Volume 71, Number 1, January-February 2019
Siriraj Medical Journal
SMJ
ISSN 2629-995X
ORIGINAL ARTICLE
1 The Effect of Kao-Ta (9-Square Step Exercise) and Kao-Ten
(9-Square Dance Exercise) on Balance Rehabilitation in
Patients with Balance Disorders
Suvajana Atipas, et al.
8 Correlation between Hypo-osmotic Swelling Test and DNA
Fragmentation Assessed by the TUNEL Assay in
Asthenozoospermia
Pitak Laokirkkiat, et al.
14 Lipid-Poor Adrenal Lesion: Differentiation of Benign from
Malignant Disease by Using Imaging Features on Routine
Contrast-Enhanced CT
Shanigarn Thiravit, et al.
21 Normative Values of Second-Trimester Maternal Serum
Markers Using an Automated Assay Platform for Down
Syndrome Screening
Kusol Russameecharoen, et al.
25 Economic Evaluation of Ready-to-use Injectable
Medications by Pharmacy Department Compared with
the Traditional System of Individual Preparation by Nurse
Prapaporn Noparatayaporn, et al.
31 Administration of Renin-Angiotensin System Inhibitor
Affects Tumor Recurrence and Progression in Non-Muscle
Invasive Bladder Cancer Patients
Saran Maneesuwansin, et al.
38 Cancer Pain Management: Is It Still Problematic?
Pramote Euasobhon, et al.
44 Patient Interviews Improve Empathy Levels of Preclinical
Medical Students
Sapol Thepwiwatjit, et al..
52 Effect of Intraoperative Hypothermia on Surgical
Outcomes after Colorectal Surgery within an Enhanced
Recovery after Surgery Pathway
Varut Lohsiriwat, Panumat Jaturanon
59 A Randomized Controlled Trial of the Correlation between
Iodine Supplement in Pregnancy and Maternal Urine
Iodine and Neonatal Thyroid Stimulating Hormone Levels
Saifon Chawanpaiboon, Vittaya Titapant
66 The Implementation of a Red Blood Cell Transfusion
Guideline in Critically III Surgical Patients at Siriraj Hospital
Anticha Siritongtaworn, et al.
74 Prevalence of and Factors Associated with Inappropriate
Indications for Transthoracic Echocardiography in Adult
Outpatients at Siriraj Hospital
Kesaree Punlee, et al.
80 Esophageal Replacement in Children: A 10-Year,
Single-Center Experience
Mongkol Laohapensang, et al.
REVIEW ARTICLE
89 Extracellular Vesicles in Malaria Infection
Ladawan Khowawisetsut, et al.
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By Pitak Laokirkkiat, et al.
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Volume 71, No.1: 2019 Siriraj Medical Journal
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1
Original Article
SMJ
Suvajana Atipas, M.D.*, Cheerasook Chongkolwatana, M.D.*, Thitaree Suwannutsiri, M.D.**, M.L.
Kanthong ongyai, M.D.*, Supaporn Henggrathock, B.ATM.***, Pravit Akarasereenont, M.D., Ph.D.***
*Department of Otorhinolaryngology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok 10700, **Otorhinolaryngology Unit, Taksin
Hospital, Bangkok 10600, ***Center of Applied ai Traditional Medicine, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok 10700,
ailand.
Effect of Kao-Ta (9-Square Step Exercise) and
Kao-Ten (9-Square Dance Exercise) on Balance
Rehabilitation in Patients with Balance Disorders
Corresponding author: Cheerasook Chongkolwatana
E-mail: cheerasook.cho@mahidol.ac.th
Received 13 June 2018 Revised 28 September 2018 Accepted 7 November 2018
ORCID ID: 0000-0002-8423-6357
doi: http://dx.doi.org/10.33192/Smj.2019.01
ABSTRACT
Objective: To study the eect of Kao-ta (9-square step exercise) and Kao-ten (9-square dance exercise) on balance
improvement in patients with balance disorders.
Methods: is prospective pilot study in patients with balance disorders was conducted at the outpatient clinic,
Department of Otorhinolaryngology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, ailand
from December 2015 to December 2016. Patients diagnosed by clinical symptoms and at least one abnormal
condition on posturography were taught how to perform Kao-ta and Kao-ten exercise. Participants were provided
with the equipment necessary to create a nine square grid at home. ey were instructed to perform 3 minutes of
Kao-ta followed by 2 minutes of Kao-ten twice per day for at least 45 days in an 8-week period. Posturography and
visual analogue scale (VAS) of balance symptom severity were compared between before and aer exercise program.
Results: Eleven patients with balance disorders were included. e mean age was 57.2±12.9 years (range: 33-70), and
all patients were women. e average composite equilibrium score at baseline was 64.4±8.1. Aer 8 weeks of Kao-ta
and Kao-ten, the average composite equilibrium score increased to 73.8±10.2 (p<0.01). e median (P25, P75) of
the abnormal equilibrium score condition decreased from 2 (1, 3) at baseline to 1 (0, 2) aer 8 weeks (p=0.016). e
median VAS of balance symptom severity decreased from 4 (3, 6) at baseline to 2 (0.2, 5.5) aer 8 weeks (p=0.028).
Conclusion: Kao-ta and Kao-ten exercise can improve symptoms in patients with balance disorders aer 8 weeks
of exercise
Keywords: Kao-ta; Kao-ten; balance rehabilitation; patients; balance disorders; ai traditional medicine (Siriraj
Med J 2019;71: 1-7)
INTRODUCTION
Balance disorders can be found in any age group,
but they are more commonly observed in older adults.
e prevalence of balance disorder complaints is 5-10%
of patients among general practitioners, and 10-20%
of patients among otorhinolaryngologists.
1
During
2014-2016, about 12% of the patients (7,250 patients)
that visited the outpatient clinic of the Department of
Otorhinolaryngology, Faculty of Medicine Siriraj Hospital
had balance disorder, and 48.12% of those were aged
greater than 60 years.
Balance requires coordination among the visual,
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2
proprioceptive, and inner ear vestibular systems. e
central nervous system receives inputs from these systems,
and then sends back information to eectuate muscle
control of the eyes, neck, torso, and extremities to maintain
balance. Any impairment along any of these pathways
can cause balance disorders and/or vertigo.
Treatments of balance disorders and vertigo include:
1. Specic treatment, such as canalith repositioning
for benign paroxysmal positional vertigo. It should be
noted that many diseases have no specic treatment,
and some diseases are incurable.
2. Symptomatic treatment, which consists mostly
of medication therapy
3. Balance rehabilitation
When balance disorders occur, the vestibular
system will initiate compensatory adaptations in an
eort to correct the aberration. ese adaptations and
recovery of defective functions can take days to weeks.
However, balance may not fully recover in all patients,
and rehabilitation plays a key role in the recovery of
balance in this group.
e goal of rehabilitation is to improve overall
body balance. Cawthorne-Cooksey exercise, which was
introduced in 1940, is one of the most well-known imbalance
rehabilitation methods.
2
is exercise is indicated in
patients with prolonged symptoms or partial recovery.
Cawthorne-Cooksey exercise can improve balance in
up to 50-80% of patients
3-5
, and it has demonstrated
benet in patients with acute vertigo. Other traditional
exercises, such as Tai chi
5-7
and Wii Fit
8
, have shown
rehabilitation benet in patients with balance disorders.
A 2008 study in aquatic physiotherapy using whirlpool
as part of an exercise protocol revealed positive eects
on unilateral vestibular hypofunction.
9
Kao-ta (9-square step exercise) or Ouay’s Test is
a form of exercise invented in 1970 by Professor Dr.
Ouay Ketusinh, a renowned Professor of Physiology
from the Faculty of Medicine Siriraj Hospital, Mahidol
University. Professor Dr. Ouay Ketusinh also founded a
ai traditional medical school that was later named the
Center of Applied ai Traditional Medicine, Faculty of
Medicine Siriraj Hospital, Mahidol University, Bangkok,
ailand. In 1972, he presented his invention Nine Square
Test or Ouay’s Test to the International Committee on
the Standardization of Physical Fitness Test (ICSPFT) in
Cologne, Germany. He later presented his Nine Square
Health Twist exercise (original name of the 9-square
dance exercise) at the Olympic Conference in the same
year. He published his article describing Kao-ta and
Kao-ten in ai language in 1984.
10
Kao-ta was originally employed as a speed test.
Kao-ten (9-square dance exercise), which was developed
from Kao-ta, requires more coordination than Kao-ta
because it includes many body turns.
11
A user of either
exercise maneuvers his/her body within a square somehow
drawn or represented on the oor or ground. e box,
which can have overall dimensions of 90 x 90 cm, 120
x 120 cm, or 150 x 150 cm, is divided into 9 equal sized
boxes – 3 at the bottom, 3 at the middle, and 3 at the
top (Fig 1 and 2). Both exercises are safe and easy to
perform without elaborate equipment, and both are
health promotion techniques that are taught in ai
traditional medicine. Although another original goal
of these exercises was to strengthen the cardiovascular
system among older adults, the fact that both involve head
turning and body balance indicates that they are rooted
in the same principles as other balance improvement
exercises.
e aim of this rst ever pilot study was to investigate
the ecacy of Kao-ta and Kao-ten ai exercises for
improving symptoms in patients with balance disorders.
Subjective evaluation using VAS symptom score, and
objective evaluation using posturography score were
compared between before and aer the prescribed 8-week
ai exercise program.
MATERIALS AND METHODS
is prospective pilot study in patients with balance
disorders was conducted at the outpatient clinic of the
Department of Otorhinolaryngology, Faculty of Medicine
Siriraj Hospital, Mahidol University, Bangkok, ailand
during the December 2015 to December 2016 study period.
Siriraj Hospital is ailand’s largest national tertiary
referral center. Patients aged 18-70 years with persistent
and prolonged (more than 4 weeks) balance disorder
were enrolled. Balance disorders included vestibular
neuritis, dizziness in the elderly or vestibulopathy, and
inability to maintain balance in at least one (out of six)
condition on posturography. Patients having one or
more of the following were excluded: inability to properly
or adequately perform Kao-ta and/or Kao-ten (e.g.,
neuromuscular disorder, vision defect); having central
cause of balance disorders; having psychiatric problems;
and/or, having disease with specic treatment (e.g.,
benign paroxysmal positional vertigo). e protocol
for this study as approved by the Siriraj Institutional
Review Board (COA no. SI 704/2015), and all included
patients provided written informed consent.
e objective evaluation of balance was performed using
SMART Equitest® Computerized Dynamic Posturography
(NeuroCom International, Inc., Clackamas, OR, USA).
Briey, patients stand on a support surface that can be
Atipas et al.
Volume 71, No.1: 2019 Siriraj Medical Journal
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3
Original Article
SMJ
Fig 1. Kao-ta (9-square step exercise)
Preparatory position: Stand with both feet within the bottom le square (1a).
Exercise steps: Moving in a counterclockwise direction, move your right foot directly to the right into the bottom right square (1b), followed
then by your le foot. Now both of your feet are once again in the same square (1c). Now move your right foot straight ahead to the top
right square (1d), followed then by your le foot (1e). Now move your le foot directly to the le into the top le square (1f), followed then
by your right foot (1g). Now, move your le foot directly backwards to the bottom le square, which is where you rst started (1h), followed
then by your right foot (1i). Now that you have completed one cycle of the exercise, move to the bottom right square to begin, except now
you will repeat these movements going in the opposite (clockwise) direction.
Fig 2. Kao-ten (9-square dance
exercise)
Preparatory position: Stand with
your feet apart, with your le foot
in the bottom le square, and your
right foot in the bottom right square
(2a).
Exercise steps: Move your le foot
diagonally to the top right square
(2b). Now bring your right leg around
the front of your le leg, and position
your right foot in the top le square
(2c). Now move your le foot back
to the bottom le square where you
started with your le foot (2d), followed
by movement of your right foot to
the bottom right square where you
started with your right foot (2e).
en you will repeat these movements
going in the opposite direction.
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4
controlled to be level or tilted. Located in front of the
patient is a visual eld that can be stable or moved in and
out. e result of this test is the “equilibrium score”, which
is an average of body balance in 6 dierent conditions.
Persons with normal balance have normal results in
all conditions. e result is also reported as how many
of the equilibrium conditions are abnormal out of the
six conditions. e program then uses the equilibrium
score and the degrees of sway to calculate a “composite
equilibrium score”, which is a weighted average of all
6 tested conditions. An improvement in the composite
equilibrium score of greater than 2 standard deviations or
8 points compared to age-matched normative data within
the system is considered a meaningful improvement in
balance.
12,13
e subjective evaluation of the severity of patient
imbalance was performed using a visual analogue scale
(VAS). Patients rated the intensity of their imbalance
using a 10 cm VAS, with a 0 indicating no symptoms of
imbalance, and a 10 indicating the worst possible level
of imbalance.
14
Patients were taught how to correctly perform the
Kao-ta and Kao-ten exercises by instructors from the
Center of Applied ai Traditional Medicine, Faculty
of Medicine Siriraj Hospital, Mahidol University. Each
participant was also given an instruction manual (Fig 1
and 2), a music CD (with a rhythm and beat matched to
the steps of the exercises), and corrugated plastic sheets
with a roll of colored adhesive tape that were together used
to make a nine square grid. A video clip of the Kao-ta and
Kao-ten exercises is available at http://www.si.mahidol.
ac.th/ent/knowledge/videos/kao-ta_kao-ten_exercise.
php.
Patients had to perform the Kao-ta and Kao-ten
exercises to music twice a day for a total of 5 minutes per
session. Each session consisted of Kao-ta for 3 minutes,
followed by Kao-ten for 2 minutes. Each patient had to
perform at least 45 days of exercise within the 8-week study
period (80% of days). Since balance disorder patients are
at higher risk for falling, all study participants were asked
to perform their exercises in their bare feet. No socks or
slippers were allowed. Moreover, it was recommended
that a caretaker participate as an observer during each
exercise session in case of a fall or some other unexpected
event. Patients were advised to take a dimenhydrinate
tablet as rescue medication if severe symptoms developed.
Patients were instructed to record the time and date of
their exercises, and any adverse events in a logbook that
was provided to each study participant.
e results of posturography testing and VAS scores
of all patients were recorded at baseline and aer the
8-week exercise program. Patients were asked to return
to the clinic for a follow-up visit at 4 weeks so that we
could check their logbooks, inquire about adverse events,
evaluate patient satisfaction, and ensure that the exercises
were being performed correctly.
Statistical analysis
PASW Statistics for Windows version 18.0 (SPSS,
Inc., Chicago, IL, USA) was used to perform all statistical
analyses. Descriptive statistics are reported as mean ±
standard deviation, number and percentage, or median
(P25, P75). Paired t-test was used to compare composite
equilibrium scores, and Wilcoxon signed-rank test was
used to compare equilibrium scores and VAS symptom
scores between baseline and aer 8 weeks of exercise. A
p-value less than 0.05 was regarded as being statistically
signicant.
RESULTS
Of the 12 patients that initially enrolled, one patient
was not able to complete the study due to imbalance
symptoms that were too severe to perform Kao-ta and
Kao-ten. e remaining 11 patients completed the study
and were included in the nal analysis. e mean age of
patients was 57.2±12.9 years (range: 33-70), and all of
them were women. e diagnoses of study participants
were, as follows: nonspecic dizziness (7 patients, 63.6%),
vestibulopathy (3 patients, 27.3%), and probably Meniere’s
disease (1 patient, 9.1%) (Table 1).
Posturography
e average composite equilibrium score at baseline
was 64.4±8.1. Aer 8 weeks of Kao-ta and Kao-ten, the
mean±standard deviation composite equilibrium score
increased to 73.8±10.2 (p<0.01). e median (P25, P75) of
abnormal equilibrium score condition decreased from 2
(1, 3) at baseline to 1 (0, 2) aer 8 weeks (p=0.016). Aer
8 weeks of exercise, 10 patients (90.9%) had improvement
in their composite equilibrium score, and 6 of them
(54.5%) had scores that increased by at least 8 points.
Seven of 11 patients (63.6%) had at least one condition
that returned to normal aer completion of the 8-week
exercise program (Tables 2 and 3).
Visual analogue scale (VAS)
e median (p25, P75) VAS of balance symptom
severity decreased from 4 (3, 6) at baseline to 2 (0.2, 5.5)
aer 8 weeks (p=0.028). Nine patients (81.8%) rated their
severity of imbalance as improved (Tables 2 and 3). Four
patients (36.6%) were unable to complete a full 5 minutes
of exercise during the rst one or two days; however,
Atipas et al.
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5
Original Article
SMJ
TABLE 1. Demographic characteristics and clinical diagnosis of the 11 study participants.
Age (years), mean±SD (range) 57.2 ± 12.9 (range: 33-70)
Gender, n (%)
Male 0 (0.0%)
Female 11 (100%)
Diagnosis, n (%)
Nonspecicdizziness 7(63.6%)
Vestibulopathy 3 (27.3%)
Probable Meniere’s disease 1 (9.1%)
Abbreviation: SD=standard deviation
TABLE 2. Clinical diagnosis and result of objective test and subjective VAS symptom score.
Patient Age (yr) Diagnosis Composite Equilibrium score VAS
equilibrium score (number of abnormal
conditions)
Pre Post Pre Post Pre Post
1 51 ProbableMeniere'sdisease 43 50 5 5 6.8 6
2 69 Vestibulopathy 66 72 2 1
†
4.5 5.5
3 70 Vestibulopathy 65 73* 1 1 3 1
4 33 Vestibulopathy 67 85* 3 0
†
3 2
5 65 Nonspecicdizziness 69 76 2 1
†
7.5 5.5
6 67 Nonspecicdizziness 70 70 2 2 4 0.2
7 68 Nonspecicdizziness 69 78* 1 0
†
5 3
8 42 Nonspecicdizziness 71 84* 1 0
†
4 1
9 58 Nonspecicdizziness 68 81* 2 0
†
6 0
10 43 Nonspecicdizziness 63 80* 2 0
†
0 0
11 63 Nonspecicdizziness 57 63 3 3 3 4
*Composite equilibrium score increased ≥8 points
†
Abnormal equilibrium score that returned to normal in at least 1 condition
Abbreviations: VAS=visual analog scale; Pre=before exercise; Post=aer 8 weeks of exercise
TABLE 3. Comparison of composite equilibrium score, abnormal equilibrium score condition, visual analogue
score (VAS) before and aer 8 weeks of exercise.
Before exercise After exercise P-value
Compositeequilibriumscore 64.4±8.1 73.8±10.2 <0.01
Abnormalequilibriumscorecondition* 2(1,3) 1(0,2) 0.016
VAS 4(3,6) 2(0.2,5.5) 0.028
Data are presented as mean ± standard deviation or median (P25, P75)
A p-value<0.05 indicates statistical signicance
*Abnormal equilibrium score condition indicates the median (range) number of the total of 6 conditions that had an abnormal equilibrium
score at baseline and aer 8 weeks of exercise
Volume 71, No.1: 2019 Siriraj Medical Journal
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6
those patients developed the stamina to exercise for a
full 5 minutes in all subsequent exercise sessions. No
patients had to take rescue medication, and no adverse
events were reported or observed in this study.
DISCUSSION
Balance rehabilitation is one of the most eective
methods for treating balance disorders. Kao-ta and
Kao-ten ai exercises, which were invented by Professor
Dr. Ouay Ketusinh, were designed to improve coordination
among the eyes, head, torso, and extremities. e design
features of Kao-ta and Kao-ten inspired this research
team to investigate these exercises for balance benet
in patients with balance disorders.
Aer 8 weeks of Kao-ta and Kao-ten exercise in
this study, 90.9% of patients had a better composite
equilibrium score, and 54.5% of those had an increase
at least 8 points, which indicates that they had better
balance.
12,13
Moreover, the equilibrium score of at least
one condition returned to normal in 63.6% of patients.
Nine patients (81.8%) reported improved balance. ese
results are comparable to those observed aer Cawthorne-
Cooksey exercise in ai elderly population with imbalance
disorders.
4
Cawthorne-Cooksey exercise was reported
to eectuate 50-80% improvement in patient balance.
3-5
Compensation usually occurs 2-3 days aer symptoms
of balance disorders develop, but 30% of patients do not
compensate well enough.
15
erefore, we only recruited
patients with symptoms for 4 weeks or more in order
to exclude patients that might develop spontaneous
compensation.
All patients in our study were female, so there were
no gender or physical strength biases. We also endeavored
to reduce the probability of incorrect exercise technique
by inviting instructors from the Center of Applied ai
Traditional Medicine, Faculty of Medicine Siriraj Hospital,
Mahidol University to teach correct Kao-ta and Kao-ten
technique to our study patients. A logbook was given to
each patient to record daily exercise times, and to note
any accidents or adverse events that occurred during
the study period. In addition, all patients attended our
outpatient clinic at 4 weeks for a follow-up to assess
patient satisfaction and to inquire about adverse events.
During that visit, patient logbooks were checked, and
questions were asked to elicit information specic to
the correctness and regularity of the 2 prescribed ai
exercises.
e results of this pilot study revealed that Kao-ta
and Kao-ten exercise can signicantly improve patient
balance. However, a controlled study in a larger study
population is needed conrm our ndings, and to further
elucidate the scope of the benet conferred by these ai
exercises. Comparatives studies that compare Kao-ta and
Kao-ten with other balance rehabilitation methods are
also recommended.
Limitations
is pilot study has a mentionable limitation. Patients
were required to perform at least 45 days of exercise
(80%) during the 8-week study period. It is, therefore,
possible that patients that performed more than 45
days of exercise may have realized better outcomes than
patients that performed only the minimum 45 days of
exercise. No provision was made in this study to evaluate
performance based on the number of days of exercise
beyond the 45-day minimum.
CONCLUSION
e results of the rst ever pilot study revealed that
Kao-ta and Kao-ten exercise can improve symptoms in
patients with balance disorders aer 8 weeks of exercise
when evaluated by visual analogue scale and computerized
dynamic posturography. No exercise-related adverse
events were observed or reported.
ACKNOWLEDGMENTS
e authors gratefully acknowledge Mr. Suthipol
Udompunturak of the Division of Clinical Epidemiology,
Department of Research and Development, Faculty of
Medicine Siriraj Hospital, Mahidol University for assistance
with statistical analysis; Dr. Premyot Ngaotepprutaram
of the Department of Otorhinolaryngology, Faculty of
Medicine Siriraj Hospital, Mahidol University for his
contributions to this study; Ms. Narisara Nateluecha and
Ms. Sukritta Pongsitthichok of the Center of Applied
ai Traditional Medicine, Faculty of Medicine Siriraj
Hospital, Mahidol University for their contributions to
this study; and, Mrs. Jeerapa Kerdnoppakhun for her
assistance with manuscript development.
Conict of interest declaration
All authors declare no personal or professional
conicts of interest relating to any aspect of this study.
Funding disclosure
is study was funded by a grant from the Faculty of
Medicine Siriraj Hospital, Mahidol University, Bangkok,
ailand (grant no. R15931030).
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vestibular vertigo: a neurotological survey of the general
population. Neurology. 2005;65:898-904.
Atipas et al.
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2. Anne Shumway-Cook PE. Vestibular rehabilitation-An eective,
evidence-based treatment [January 17, 2015]. Available from:
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3. Hecker HC, Haug CO, Herndon JW. Treatment of the vertiginous
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4. Prasansuk S, Siriyananda C, Na Nakorn A, Atipas S, Chongvisal
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5. Wrisley DM, Pavlou M. Physical therapy for balance disorders.
Neurol Clin. 2005;23:855-74, vii-viii.
6. Millar JL, Schubert MC, Shepard NT. Vestibular and balance
rehabilitation: Program essentials. In: Flint PW, Haughey BH,
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MM, editor. Cummings otolaryngology-head and neck surgery.
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M. Vestibular compensation and vestibular rehabilitation.
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49-57.
8. Cone BL, Levy SS, Goble DJ. Wii Fit exer-game training
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young adults. Gait Posture. 2015;41:711-5.
9. Gabilan YP, Perracini MR, Munhoz MS, Gananc FF. Aquatic
physiotherapy for vestibular rehabilitation in patients with
unilateral vestibular hypofunction: exploratory prospective
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10. Ketusinh O. Kao-Ta, Kao-Ten, Kao Ma, Kao Sung in exercise.
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Medicine. 9-Square Step Exercise. In: Tawee Laohapand,
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Supavanich Press; 2014. p. 67-70.
12. NeuroCom international I. Clinical interpretation guide
computerized dynamic posturography. USA: NeuroCom
international, Inc.; n.d. p. 37-52.
13. Wrisley DM, Stephens MJ, Mosley S, Wojnowski A, Duy
J. Burkard R. Learning eects of repetitive administrations
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14. Kammerlind AS, Hakansson JK, Skogsberg MC. Eects of
balance training in elderly people with nonperipheral vertigo
and unsteadiness. Clin Rehabil. 2001;15:463-70.
15. Teggi R, Caldirola D, Fabiano B, Recanati P, Bussi M. Rehabilitation
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8
Pitak Laokirkkiat, M.D.*, Nida Jareemit, M.D.*, Isarin anaboonyawat, M.D., MMedSci.*, Hollie Smith,
MMedSci.**, Sukanya Sriiam, M.Sc.*
*Infertility unit, Department of Obstetrics and Gynecology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok 10700, ailand,
**Nurture Fertility, e East Midlands Fertility Clinic, Nottingham, United Kingdom.
Correlation between the Hypoosmotic SwellingT
and DNA Fragmentation Assessed by the TUNEL
Assay in Asthenozoospermia
Corresponding author: Isarin anaboonyawat
E-mail: isarin.tha@mahidol.ac.th
Received 4 June 2018 Revised 9 October 2018 Accepted 7 November 2018
ORCID ID: 0000-0002-3912-1382
doi: http://dx.doi.org/10.33192/Smj.2019.02
ABSTRACT
Objective: To study the correlation between the hypoosmotic swelling test (HOST) and DNA fragmentation in
asthenozoospermia assessed by the terminal deoxynucleotidyl transferase‐mediated deoxyuridine triphosphate‐
biotin nick end‐labeling (TUNEL) assay.
Methods: is cross-sectional study was conducted in 27 semen samples obtained from infertile men with
asthenozoospermia. Both HOST and TUNEL assay were performed for each sample. e sperm swelling pattern
and positive apoptosis staining of individual spermatozoa were evaluated. HOST and TUNEL scores, and the
proportion of positive staining in each grade were calculated in each sample.
Results: e results showed a negative correlation between HOST and TUNEL scores (r = -0.428, P = 0.026). Sperm
swelling grade A had a higher incidence of positive apoptosis staining when compared with other grades (P < 0.01).
ere was no statistically signicant dierence in positive apoptotic staining between other grades; nevertheless,
sperm swelling grade D tended to have a lower incidence of positive apoptosis staining.
Conclusion: Based on the results of this study, HOST may be used as an optional test to identify DNA-intact
spermatozoa whereby sperm with a grade D swelling pattern should be selected preferentially for intracytoplasmic
sperm injection (ICSI), whereas sperm with a grade A swelling pattern should be avoided for ICSI.
Keywords: Hypoosmotic swelling test; TUNEL; asthenozoospermia; male infertility; intracytoplasmic sperm
injection (Siriraj Med J 2019;71: 8-13)
INTRODUCTION
Male factor infertility constitutes 30% of infertility
causes. e World Health Organization (WHO) denes the
lower limit of normal sperm motility as 32% of progressively
motile sperm (5
th
centile, 95% condence interval [CI]:
31-34) and 40% of total motile sperm (5
th
centile, 95%
CI: 38-42).
1
e prevalence of asthenozoospermia has
been reported to be 18.71-24.19%.
2,3
In ailand, 10.78%
of infertile couples are aected by asthenozoospermia.
4
During natural human fertilization, sperm velocity is
essential for its transition through the vagina and fallopian
tubes, and for the penetration of the cumulus oophorus
and zona pellucida. erefore, sperm motility impacts
fertilization rates; i.e., low-velocity sperm are associated
with a reduced chance of zona pellucida penetration and
thereby fertilization potential.
5-7
Intracytoplasmic sperm
Laokirkkiat et al.
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injection (ICSI) was developed to overcome impaired
fertilization as a result of reduced semen parameters,
including asthenozoospermia. However, higher DNA
fragmentation rates are reported in asthenozoospermia
and the introduction of ICSI for asthenozoospermia has
promoted the transmission of disintegrated sperm DNA
to the resulting ospring, which consequently aects fetal
or postnatal development.
8
Many studies have reported
a negative correlation between sperm DNA damage and
fertilization, implantation and miscarriage rates, in addition
to embryo quality and rates of childhood diseases and
cancer.
9-14
us, it is important to discriminate DNA-
intact spermatozoa in asthenozoospermia to improve
pregnancy rates and reproductive health outcomes.
DNA integrity can be analyzed by several diagnostic
tests, such as terminal deoxynucleotidyl transferase
mediated deoxyuridine triphosphate nick end‐labeling
(TUNEL) assay.
15,16
However, drawbacks associated with
the TUNEL assay are that it is a time-consuming technique,
which requires specic skills and, most importantly, causes
sperm toxicity. erefore, the adoption of a nontoxic
sperm selection test is essential for ICSI. In cases of severe
asthenospermia, it is dicult to distinguish viable and
nonviable sperm. Since some viable sperm are known
to contain DNA-intact components, it is important
to be able to select viable sperm for ICSI to optimize
fertilization rates.
17
e hypoosmotic swelling test (HOST) is the most
commonly used method for assessing sperm vitality.
1
e basis of HOST relies on the semipermeable nature
of the sperm cell membrane, which allows the inux
of water when placed in a hypoosmotic solution, and
results in the expansion and coiling of the tail.
18
HOST
presumes that only cells with intact membranes (live
cells) will swell when within a hypotonic solution,
which allows easy identication of viable sperm.
1
It is
a simple, quick, safe, and cost-eective test. HOST was
recently introduced as a test for sperm function because
membrane integrity is important for sperm capacitation,
the acrosome reaction, and sperm–oocyte binding and
penetration. Moreover, HOST has been suggested as an
alternative test for DNA integrity. Previous studies have
demonstrated a favorable association between HOST and
many reproductive outcomes, including fertilization and
pregnancy rates.
19-21
Some studies have shown benets
from using HOST for the selection of viable nonmotile
spermatozoa.
22, 23
Casper et al. found increased fertilization
and cleavage rates (43% and 39%, respectively) when
HOST was used to select viable sperm compared with
when sperm was randomly selected in cases of complete
asthenozoospermia (26% and 23%, respectively).
24
Ortega
et al. also reported improved fertilization rates using
HOST in complete asthenozoospermia.
23
In contrast,
recent studies have failed to report a benet of HOST
utilization to select sperm on the basis of lower DNA
fragmentation levels.
20,21
However, these studies were
conducted on semen samples with parameters within the
normal range; therefore, they may not be applicable in
asthenozoospermia cases.
20,21
ere is insucient data in
relation to asthenozoospermia; therefore, we conducted this
study to clarify the correlation between HOST outcomes
and DNA fragmentation levels in asthenozoospermia as
assessed by the TUNEL assay. e secondary objective
was to identify the positive apoptosis staining in each
sperm-swelling grade.
MATERIALS AND METHODS
is cross-sectional study was approved by the
Institutional Review Board of the university hospital and
it was conducted in males aged 18 years or older who
were attending a university-based infertility unit. All
patients who requested a semen analysis and provided
written informed consent were screened. Semen samples
were collected by masturbation. Twenty-seven patients
with asthenozoospermia classied according to WHO
2010 were eligible for this study.
1
Two aliquots of each
sample were collected within 90 minutes aer semen
sample collection and liquefaction. Both aliquots were
rst prepared for HOST and then xed for TUNEL
assay. e xed and stained semen sample slides were
examined using a phase contrast microscope (BX40;
Olympus, Tokyo, Japan). A single interpreter assessed
a total of 200 spermatozoa per semen sample twice. e
semen parameters, i.e., average HOST score, average
TUNEL score, individual spermatozoa HOST grading,
individual apoptotic staining, and proportion of positive
apoptosis staining in each grade of sperm swelling, were
calculated and recorded.
HOST
According to WHO 2010,
1
the hypoosmotic solution
was prepared by dissolving 0.375 g of sodium citrate
dehydrate and 1.351 g of D-fructose in 100 mL of puried
water. An aliquot of 100 mL of semen was mixed with
1 mL of the swelling solution before being incubated
at 37°C for 30 minutes. Ten microliters of the mixed
sample was then placed on a clean slide and covered
with a coverslip. In live sperm with normal membrane
function, the hypoosmotic buer diused through the
membrane into the sperm tail. Under microscopy, live
spermatozoa showed various degrees of tail swelling
and curling, whereas dead cells exhibited no membrane
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10
changes. All membrane-change patterns were categorized
from grades A to G (Fig 1), where grade A corresponds
to no tail swelling. e HOST score was obtained from
the average percentage of total swollen spermatozoa
(excluding the natural swelling before treatment with
hypoosmotic buer). e lower reference limit for normal
vitality was 58% (5
th
centile, 95% CI: 55-63).
1
TUNEL assay
Both semen slides were fixed after HOST. The
TUNEL assay was then performed according to the
manufacturer’s instructions for ApopTag® Plus Peroxidase
In Situ Apoptosis Detection Kit (Merck, Kenilworth, NJ,
USA). In the assay, spermatozoa with DNA fragmentation
stained brown, while sperm containing intact DNA
stained green (Fig 2).
Fig 1. Aer incubation in hypo-osmotic solution for 30 minutes, every sperm was assessed under phase-contrast microscopy and categorized
in 7 groups, from grade A to G as demonstrated in the gure. Grade A showed the sperm with no membrane swelling which indicated a
non-viable sperm. Grade B to G showed dierent grades of tail swelling in viable sperms.
e TUNEL score was calculated using the formula:
% Average positive staining =
Average of stained apoptotic spermatozoa
x 100
200
e proportion of positive apoptosis staining in each grade of sperm swelling was calculated as follows:
Positive apoptosis staining (%) =
number of positive apoptosis staining in that grade
x 100
total number of spermatozoa in that grade
Statistical analysis
e semen parameters were presented as mean,
median, and standard deviation. e intraclass correlation
coecient was used to evaluate consistency in two time
measurements for the HOST and TUNEL scores in each
sample. e correlations between HOST and TUNEL
scores were analyzed using Pearson correlation and
Wilcoxon signed-rank tests. P < 0.05 was accepted as
statistically signicant. Data were analyzed using PASW
Statistics (v. 18.0; SPSS Inc., Chicago, IL, USA).
RESULTS
e characteristics of 27 semen samples are shown in
Table 1. A fairly negative correlation between HOST and
TUNEL scores (r = -0.428, P = 0.026) was revealed using
Pearson correlation (Fig 3). ere was high agreement in
A C EB D F G
Fig 2. Apoptosis staining under normal light microscopy was
demonstrated in the gure.
e positive apoptosis staining was demonstrated in the
solid circle.
e negative apoptosis staining was displayed in the circle
of dotted line.
Laokirkkiat et al.
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TABLE 1. Characteristics of semen parameters (N = 27) were displayed.
Semen parameters Mean ± SD
pH 7.7 ± 0.3
Volume (mL) 2.5 ± 1.8
Sperm concentration (10
6
/mL) 21.4 ± 15.1
Total sperm count (10
6
/mL) 51.9 ± 43.7
Progressive motility (%) 24.1 ± 5.1
Vitality (%) 58.9 ± 11.3
Normal morphology (%) 8.9 ± 5.9
Natural sperm swelling (%) 7.2 ± 3.2
r = - 0.428
p = 0.026
Fig 3. Correlation between HOS TEST score and TUNEL score.
the duplicate test evaluations by the same investigator.
e intraclass correlations were 0.97 (95% CI = 0.935-
0.986) and 0.952 (95% CI = 0.898-0.978) for the HOST
and TUNEL score assessments, respectively.
When the positive staining of TUNEL was compared
among the groups with various tail swelling patterns,
grade A sperm demonstrated the highest proportion of
apoptosis staining (P < 0.01). ere was no signicant
dierence in the DNA integrity between other grades.
Nevertheless, the sperm swelling grade D tended to have
a lower proportion of positive apoptosis staining (Fig 4).
DISCUSSION
e quality of sperm is highly associated with the
outcome of assisted reproductive technology. e selection
of sperm with high DNA integrity is an important step
prior to ICSI to optimize the outcome because DNA
Fig 4. Proportion of positive apoptosis staining in each grade of
sperm swelling.
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12
fragmentation is known to aect fertilization rates and
embryo development.25 Assessment based solely on
morphology or vitality alone may not be enough because
many sperm with normal morphology and motility
still contain damaged genetic material, especially in
asthenospermia.
8
Membrane function contributes a key role in various
sperm function competencies, i.e., capacitation, the acrosome
reaction, and fertilization. Membrane disintegration
is suggested to be associated with implantation failure
and miscarriage.
26
HOST is a vitality test used to reveal
the sperm membrane integrity and it is not only useful
in increasing fertilization rates in cases of complete
asthenospermia, but can also be used to assess sperm
DNA fragmentation.
19,20
is study demonstrates the relationship between
HOST and TUNEL assay in cases of asthenozoospermia
and highlights the benets of using HOST to select sperm
with lower levels of DNA fragmentation. e results
are consistent with the study by Stanger,
20
which also
showed a negative correlation (r = -0.81) between HOST
and TUNEL assay outcomes. However, this correlation
was stronger than in this study. is discrepancy is likely
to be the result of the dierence in study populations,
with this study enrolling only asthenozoospermic males
whose samples are prone to DNA fragmentation.
8
Stranger reported signicantly higher grade A and
lower grade D swelling patterns in abnormal semen
samples compared with normal samples.
20
However,
that study did not directly identify the DNA status in
each grade. In this study, the DNA integrity and tail
swelling pattern was assessed individually for each sperm;
therefore, the DNA integrity pattern for each grade
can be calculated. In the DNA fragmentation analysis
in each grade of sperm swelling, grade A was found to
contain the highest portion of DNA fragmentation,
while grade D obtained the lowest percentage. In the
remaining viable sperm, healthy DNA was identied
mostly in grade C followed by grades E, F, B, and G
(Fig 4), which concurs with previous studies that suggested
a correlation between tail swelling pattern and DNA
damage.
20,21
ese results are also consistent with a study
conducted in males with normal semen parameters.
27
e HOST or hypotonic resistance test distinguishes the
sperm with dierent capacities in the function of Na
+
/
K
+
and Na
+
/H
+
exchange of the membrane. Sperm that
exhibited the minimal swelling patterns, as in grade D,
are supposed to contain higher membrane competency
along the agellum.
28
Nevertheless, a high level of swelling
signies impaired Na
+
/K
+
ATPase function.
20
e normal
HOST-sensitive membrane feature is suggested to be
linked with less or no DNA damage.
20
erefore, this
nding suggests that HOST is clinically applicable for
selecting better-quality sperm with a grade D swelling
pattern for ICSI.
CONCLUSION
There is a fairly negative correlation between
HOST outcomes and DNA fragmentation levels in
asthenozoospermia as assessed by the TUNEL assay.
e study results support the use of HOST as a tool
to identify viable spermatozoa in terms of intact DNA
in asthenozoospermia to improve reproductive health
outcomes. As sperm swelling grade A has the highest
incidence of positive apoptosis staining when compared
with other grades, they should not be selected for ICSI,
whereas grade D tend to have a lower incidence of positive
apoptosis staining and therefore should be prioritized
for selection during ICSI.
ACKNOWLEDGMENTS
is study was nancially supported by the Siriraj
Grant for Research Development, Faculty of Medicine
Siriraj Hospital, Mahidol University (Grant Number
R015532035). e authors thank Miss Julaporn Pooliam
and Dr. Ratikorn Saejong for their assistance in performing
the statistical analysis.
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Shanigarn Thiravit, M.D.*, Natee Ruangpaisanbamrung, M.D.**, Voraparee Suvannarerg, M.D.*, Phakphoom Thiravit, M.D.*
*Department of Radiology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok 10700, **Bangkok Hospital Chanthaburi, Chanthaburi
22000, ailand.
Lipid-Poor Adrenal Lesion: Differentiation of Benign
from Malignant Disease by Using Imaging Features
on Routine Contrast-Enhanced CT
Corresponding author: Phakphoom iravit
E-mail: art.phak@gmail.com
Received 24 May 2018 Revised 24 September 2018 Accepted 7 November 2018
ORCID ID: 0000-0001-5058-9624
doi: http://dx.doi.org/10.33192/Smj.2019.03
ABSTRACT
Objective: To assess the eectiveness of CT imaging features of lipid-poor adrenal lesions on routine contrast-
enhanced CTs in dierentiating benign from malignant masses.
Methods: A retrospective study was performed on 84 lipid-poor adrenal lesions (HU >10 on unenhanced CT scans),
which were sized 1-4 cm, had a proven nal diagnosis, and were detected during routine contrast-enhanced MDCT
studies. Of those, 58 were found in patients with an underlying extra-adrenal malignancy. Two authors determined
the morphological features according to their shape, margin, density on unenhanced images, and enhancement
pattern. e sensitivity, specicity, and positive and negative predictive values were also calculated for each feature
which suggested benignancy, plus a combination of those features.
Results: ere were 46 (55%) benign and 38 (45%) malignant adrenal masses. e low-density feature (10-20
HU on unenhanced CT images) indicated as benign with a high specicity of 92%, even in patients with known
malignancy. e other features (round/oval shape, smooth margin, and homogenous enhancement) showed a high
sensitivity (75%-85%) but a low specicity (39%-56%) in predicting benignity. e combined features for presumed
benignancy could predict a benign mass with the highest specicity of 95%.
Conclusion: e small, lipid-poor, adrenal masses detected by routine contrast-enhanced CTs are likely to be benign
when their internal density on unenhanced images is not higher than 20 HU and/or, especially, when a combination
of all morphological features for presumed benignancy presents.
Keywords: Adrenal adenoma; adrenal gland; CT (Siriraj Med J 2019;71: 14-20)
INTRODUCTION
The prevalence of an incidental adrenal mass
discovered on chest or abdominal contrast-enhanced
CT scans is approximately 4% which is mainly a benign
adenoma.
1
Using internal attenuation of ≤ 10 Hounseld
units (HU) on unenhanced CT images, sensitivity and
specicity for the diagnosis of adrenal adenoma were
71% and 98%, respectively.
2,3
Previous study reported
that approximately 30% of non-fat-containing adrenal
masses (HU > 10) were lipid-poor adrenal adenomas.
2
Currently, an adrenal gland CT with a 10- or 15-minute
delayed protocol is usually performed to evaluate lipid-
poor adrenal lesions. A diagnosis of lipid-poor adenoma
is established by calculating the absolute contrast washout
(ACW) or relative contrast washout (RCW) values. e
thresholds for ACW and RCW for 10-minute delayed
Thiravit et al.
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protocol were 37% and 53%, respectively - with the
sensitivity and specicity of 100% and 98%, respectively.
4,5,6
Despite the high sensitivity and specificity of
specic CT adrenal protocol, this protocol requires
another appointment, more expense and exposure to
radiation and contrast media.
7
ere have been eorts
to dierentiate benign from malignant adrenal masses
by using the morphological features apparent on the
rst routine CT scan.
7-10
One study found that benign
adrenal masses were associated with homogeneous
low attenuation, an enlarged gland with the adrenal
conguration maintained, a round or oval shape, and
a thin or absent rim enhancement, whereas malignant
masses were associated with a size exceeding 4 cm, a thick
or nodular enhancing rim, and adjacent organ invasion.
9
Another study showed that an irregular margin and a
thick rim enhancement were highly associated with
malignancy but had low sensitivity.
7
erefore, the aim
of this study was to evaluate the CT imaging features of
lipid-poor adrenal lesions on routine contrast-enhanced
CTs to dierentiate benign from malignant masses.
MATERIALS AND METHODS
Subjects
is retrospective study was approved by the Siriraj
Institutional Review Board (Si 588/2015). We identied
175 patients with lipid-poor adrenal lesions detected on
routine contrast-enhanced CTs of the chest or abdomen
at our hospital between January 2013 and March 2014.
A lipid-poor adrenal lesion was dened as any adrenal
lesion with an HU level > 10 on an unenhanced CT scan.
We excluded those patients who had adrenal masses sized
smaller than 1 cm or larger than 4 cm, had no accessible
pathological reports, or had had no follow-up studies
for more than 1 year. Per exclusion criteria, 78 patients
with 84 proven lipid-poor adrenal masses (1-4 cm) le
for the analysis.
Adrenal masses
e nal diagnosis of adrenal masses was conrmed
by histology or imaging studies, as follows:
A benign mass was considered if it was proven
by pathology or imaging studies (in detail, if there was a
lesion with calculated ACW and RCW values of > 53%
and/or > 37%, respectively, on a 10-minute delayed CT
adrenal protocol; or a lesion with calculated ACW and
RCW values of > 60% and/or > 40%, respectively, on a
15-minute delayed CT adrenal protocol; or a lesion with
a signal drop on opposed phase compared with in-phase
chemical shi MR images (CS-MRI); or a lesion with
1-year stability during follow-up.)
4,7
A malignant mass was considered if it was proven
by pathology or a change of nodule size, or a new lesion
developed during chemotherapeutic sessions or imaging
follow-up in patients with known malignancy; the changes
should be in keeping with primary tumors or the patient’s
conditions.
MDCT technique
Routine contrast-enhanced chest and/or abdominal
CT examinations were performed by 64-slice scanners.
e intravenous low-osmolar contrast medium was
administered at the rate of 3 ml/sec, with a dose of 100
ml for standard abdominal CTs or 80 ml for standard
chest CTs. e post-contrast images were performed 80
seconds aer contrast administration for the abdomen,
and 35 seconds aer contrast administration for chest
studies. e images were reconstructed to 1.25 mm
thickness.
Image interpretation
All lipid-poor adrenal masses on the CT images obtained
via the Picture Archiving and CommunicationSystem
(PACS) system were evaluated by 2 radiologists who had
a subspecialty in abdominal imaging and were blinded to
the nal diagnoses. e imaging features were evaluated
according to their shape (round/oval, irregular), margins
(smooth/microlobulated, irregular), densities on the
unenhanced images (10–20 HU or > 20 HU), and patterns
of enhancement (homogeneous, rim/heterogeneous). e
location, laterality, and lesion size were also recorded.
An adrenal mass was presumed to be benign if a mass
had a round or oval shape, a smooth or microlobulated
margin, a low density (10–20 HU) on an unenhanced
CT image, and a homogeneous enhancement, whereas
a malignant mass was presumed to have an irregular
shape, an irregular margin, a higher density (> 20 HU)
on unenhanced CT images, and a heterogeneous or rim
enhancement.
Statistical methods
e features presented in the benign and malignant
lesions were compared using a chi-square test, and a
P-value of < 0.05 was considered statistically signicant.
The sensitivity, specificity, positive predictive value
(PPV), and negative predictive value (NPV) for each
morphological feature for presumed benignancy and the
combined features were calculated. e corresponding
95% CIs were also reported. e statistical analyses were
performed by using SPSS Statistics for Windows, version
18.0 (SPSS Inc., Chicago, IL, USA).
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RESULTS
Patients
ere were 50 men and 28 women - with mean
age of 60 years (range 18–89). Of them, 56 (66.7%) had
a history of malignancy, while 19 (24.4%) had bilateral
adrenal masses.
Overall adrenal lesions
Of the 84 proven, lipid-poor, adrenal masses detected
on routine contrast-enhanced CT studies, 46 (55%) were
benign, and 38 (45%) were malignant masses. e nal
diagnosis of benign adrenal masses was pathologically
proven for 4 lesions (two adrenal adenomas, 1 bland
adrenal cortical tissue, and 1 granulation tissue), and by
clinical and/or imaging follow-up for another 42 lesions.
Of those 42 lesions, 17 were considered benign adenomas
according to the imaging criteria for the 10-minute delayed
CT adrenal protocol and/or CS-MRI; a further 4 lesions
were considered benign pheochromocytomas by MIBG
scans and clinical follow-up; while the remainder (23
masses) were considered benign due to lesion stability
during the 1-year follow-up.
irty-seven malignant masses were diagnosed as
metastases by either tissue diagnosis (n=1) or the imaging
criteria (n=36; nodule size changes or new nodules
developed aer chemotherapeutic session or during
imaging follow-up in patients with known malignancy).
e other was a pathologically-proven adrenocortical
carcinoma.
e mean ages of patients with benign and malignant
masses were 59.3 years and 60.2 years, respectively. e
average size of all adrenal masses was 2 cm (range 1-4
cm in maximal width). e mean sizes of the benign
and malignant masses were 18.2 and 22.0 mm, with an
SD of 7.4 and 8.2 mm, respectively (P = 0.027). Of the
adrenal masses, 59 (70%) were unilateral and 25 (30%)
bilateral. Among the 59 unilateral masses, 20 (36%)
were benign, and 36 (64%) were malignant. As for the
bilateral location, 10 out of the 25 (40%) were benign,
and the remaining 15 (60%) were malignant. ere
were no signicant dierences between the benign and
malignant masses in terms of age, sex, or laterality (P >
0.05).
A total of 56 out of the 84 (67%) lesions were found
in patients with an underlying extra-adrenal malignancy,
while the other were found in patients without one.
In the case of the 56 adrenal lesions, the masses were
signicantly malignant etiologies (n=36, 64%) rather
than benign (n=20, 36%; P < 0.001).
CT morphological features of benign and malignant
adrenal lesions
A statistically signicant dierence between benign
and malignant lesions is shown in Table 1. Most benign
adrenal masses had a round/oval shape (91%), a smooth/
microlobulated margin (93%), and a homogenous
enhancement (82%); those three features showed a very
high sensitivity but rather low specicity to dierentiate
from malignant lesions. In addition, the density of 10–20
HU on unenhanced CT images showed the highest level
of specicity (92%) among other features for presumed
benignancy. However, the detection of this feature may
be limited due to its low sensitivity (40%; Table 2).
TABLE 1. CT features of 84 lipid-poor adrenal masses.
Variable
Malignant Benign
P-value
N=38 (%) N=46 (%)
Shape Round/oval 22 (58) 42 (91) < 0.001
Irregular 16 (42) 4 (9)
Margin Smooth/microlobulated 22 (58) 43 (93) < 0.001
Irregular 16 (42) 3 (7)
Density (HU) on 10–20 3 (8) 20 (43) < 0.001
unenhanced CT images > 20 35 (92) 26 (57)
Pattern of enhancement Homogeneous 16 (42) 38 (82) < 0.001
Rim/heterogeneous 22 (58) 8 (18)
Thiravit et al.
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TABLE 2. e sensitivity, specicity, PPV, and NPV of CT features for presumed benignancy in 84 adrenal masses.
Morphology
Sensitivity Specicity PPV NPV
P-value
% (95% CI) % (95% CI) % %
Round shape 91.3 (79-98) 42.1 (26-59) 65.6 80 < 0.001
Smooth margin 93.5 (82-99) 42.1 (26-59) 66.1 84.2 < 0.001
Low density (10-20 HU) 43.5 (29-59) 92.1 (79-98) 86.9 57.3 < 0.001
Homogenous enhancement 82.6 (69-92) 57.8 (41-74) 70.4 73.3 < 0.001
Abbreviations: PPV, positive predictive value; NPV, negative predictive value
We also specically analyzed patients with a history
of malignancy (n=56), nding that the low density (10-
20 HU) showed the highest specicity (91%) in the
determination of a benign etiology, and with statistical
signicance. is result was similar to that for the patients-
without-malignancy group. However, the other features
for presumed benignancy, including a smooth margin
and homogenous enhancement, showed a high sensitivity
(75%–85%) but low specicity (42%-56%). A round/oval
shape was the only feature that showed no statistical
signicance to dierentiate between benign and malignant
diseases in this patient population (P = 0.06; Table 3).
Combined CT morphological features of benign and
malignant adrenal lesions
We combined three morphologies for presumed
benignancy (round shape, smooth margin, and homogenous
enhancement), for which the sensitivities, specicities,
PPVs, and NPVs are presented at Table 4. We also
combined all morphological features for presumed
benignancy, including the low density feature; although
this increased the specicity to 95%, it decreased the
sensitivity to 34.8%.
In predicting malignant adrenal lesions, the combination
of the high density (HU > 20) feature and a history of
malignancy showed a specicity, sensitivity, PPV, and
NPV of 86.8%, 73.9%, 73.3%, and 87.2%, respectively
(p < 0.001).
DISCUSSION
In 2017, the ACR Incidental Findings Committee
released an updated version of its White Paper on the
management of adrenal incidentalomas. ose lesions
are being detected more oen than in the past due to
the increased use of, as well as improvements to, the
spatial resolution of cross-sectional imaging modalities.
11
e guidelines in the White Paper help radiologists
and clinicians decide whether an incidentaloma should
be further investigated, followed up, or le alone. e
guidelines focus mainly on nodules of more than 1 cm
TABLE 3. e sensitivity, specicity, PPV, and NPV of CT features for presumed benignancy in 56 adrenal masses
in patients with known extra-adrenal malignancy.
Morphology
Sensitivity Specicity PPV NPV
P-value
% (95% CI) % (95% CI) % %
Round shape 85.0 (62-97) 38.9 (23-57) 43.6 82.4 0.06
Smooth margin 85.0 (62-97) 41.7 (26-59) 44.8 83.3 0.04
Low density (10-20 HU) 40.0 (19-64) 91.7 (78-98) 72.7 73.3 0.004
Homogenous enhancement 75.0 (51-91) 55.6 (38-72) 48.4 80.0 0.03
Abbreviations: PPV, positive predictive value; NPV, negative predictive value
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in size and on lipid-poor nodules (nodules with an
internal density > 10 HU on unenhanced CT scans),
both of which are usually troublesome.
11
A nodule should be evaluated whether its size is
more or less than 4 cm because size is an important
indicator for malignancy.
10,11
In the evaluation of benign
or malignant adrenal nodules, the specic adrenal CT,
CS-MRI, PET/CT, or tissue diagnosis can provide the
solution.
4,6,11
However, those do not serve as initial tools,
which consequently necessitates another appointment,
causing additional costs as well as patient worry and
inconvenience. In this study, we aimed to evaluate the CT
imaging features of lipid-poor adrenal lesions discovered
on routine contrast-enhanced CTs as those may help to
initially dierentiate benign from malignant masses.
Our study showed that there were signicant dierences
in shape, margin, density on unenhanced images and
the pattern of enhancement, to determine the nature of
lipid-poor adrenal masses. However, three morphological
features for presumed benignancy, which were a round/
oval shape, a smooth margin, and a homogeneous
enhancement, had high sensitivity but relatively low
specicity to suggest benignity. at poor specicity
implied that recognizing those imaging features on routine
CT scans cannot be helpful in dierentiating benign from
malignant adrenal diseases. e ndings were also in
concordance with the results in other studies.
7-10,12
For
example, a study by Berland et al. reported two features
for benign lesions showing 100% PPV but no statistical
signicance, namely, homogenous attenuation with little
punctuate enhancement, and a gland enlargement with the
normal conguration maintained.
9
Another interesting
nding of a homogenous attenuation lower than that
of muscle was reported with a high PPV, but it was not
investigated in our study.
9
Homogenous enhancement
is considered a feature suggesting benignancy, and it has
been reported to be seen in adrenal adenomas rather than
adrenal carcinomas, metastases, and pheochromocytomas
by many investigators.
7,10
Nevertheless, a heterogenous
enhancement may not be considered an absolute nding
for suggesting malignancy since this usually presents in
pheochromocytomas.
13
According to the widely used guidelines for adrenal
incidentalomas, a nodule can be considered as benign
by measuring the CT density with a threshold less than
10 HU.
11
Despite that, using the higher threshold value
(> 10 HU), which has been evaluated in many studies,
also demonstrates an impressive performance for lesion
characterization.
2,10,12,14
In a meta-analysis study, the
reported sensitivity and specificity for adenomas at
the cut-o values of 20 HU versus 10 HU were 88%
and 84% versus 71% and 98%, respectively.
2
Park et al.
also supported the application of a threshold value of
20 HU for characterizing an adrenal adenoma as they
reported a sensitivity, specicity, PPV, and NPV of 60%,
100%, 100%, and 67%, respectively.
14
In our study, we
showed a specicity and sensitivity of a low-density
feature (10-20 HU) on unenhanced CT images of 92%
and 44%, respectively. is was the only feature with a
high specicity among the other features for presumed
benignancy, and we also observed a similar trend in
patients either with or without known malignancy.
Despite the low sensitivity of this feature, we believe that
using a threshold of 20 HU for distinguishing benign
and malignant adrenal lesions can be acceptable, subject
to the careful consideration of some false negative rates.
In the case of patients with a history of cancer, an adrenal
nodule may be found on a routine chest or abdominal CT
during their metastatic workup. It is therefore important
to know whether an adrenal nodule is of a metastatic
or benign incidental nature. In our study, a benign-
appearing margin, shape, and enhancement pattern
were not helpful in predicting benignity except for the
low-density feature on unenhanced images, as discussed
TABLE 4. e sensitivity, specicity, PPV, and NPV of combined CT features for presumed benignancy in 84
lipid-poor adrenal masses
Combination
Sensitivity Specicity PPV NPV
P-value
% (95% CI) % (95% CI) % %
Benign A 73.9 (59-86) 65.8 (49-80) 72.3 67.6 < 0.001
Benign B 34.8 (21-50) 94.7 (82-99) 88.9 54.6 0.001
Benign A: round shape, smooth margin, and a homogenous enhancement.
Benign B: round shape, smooth margin, a homogenous enhancement, and a low density (10–20 HU).
Abbreviations: PPV, positive predictive value; NPV, negative predictive value
Thiravit et al.
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earlier. In the study by Song et al., the ndings of rim
enhancement and an irregular margin were nearly 100%
specic to malignant lesions.
7
Our results also revealed that
adrenal masses were signicantly malignant rather than
benign in patients with previously known malignancy.
As a history of malignancy is considered signicant,
diagnosis of a benign or malignant mass using the features
on a contrast-enhanced CT alone is usually not reliable
in this patient population. Further evaluation such as
CT with an adrenal protocol is recommended.
11
Due to the high sensitivity but low specicity of the
individual morphological features for presumed benignancy
to indicate a benign mass, we further combined all those
features; they included a round/oval shape, a smooth
margin, a density at 10–20 HU on unenhanced images,
and a homogeneous enhancement. is combination
was able to achieve greater percentages for specicity
(95%) and PPV (89%). is nding encourages more
condence to diagnose a lipid-poor adrenal mass as
being benign if it presents all those features for presumed
benignancy within the lesion.
Our study had several limitations. First, it was
a retrospective study with a small sample size. The
interpretation of the morphological features was also
subjective as it depended on the readers’ experiences.
Still, as this review process was close to daily practice, the
results can be applied to real-life situations. In addition,
we did not calculate the inter-observer agreement, but
instead used a consensus approach to resolve discordant
readings. Finally, there were a variety of phases of contrast-
enhanced CTs in our study. However, this did not aect
our results because dierent enhancements between
phases was not within the scope of our study.
CONCLUSION
Lipid-poor adrenal masses with a density lower than
20 HU can be considered as benign even in patients with
known malignancy. However, the rest of the individual
features for presumed benignancy have a poor performance
in dierentiating benign from malignant adrenal masses.
ACKNOWLEDGMENTS
e article was presented in form of the electronic
poster at the ECR 2018, held February 28- March 4,
2018, in Vienna, Austria. Shanigarn iravit, Voraparee
Suvannarerg, Phakphoom iravit were supported by
the Chalermphrakiat Grant from the Faculty of Medicine
Siriraj Hospital, Mahidol University.
Fig 2. A 74-year-old woman with no history of malignancy presented with signicant weight loss; bilateral adrenal masses were found on
abdominal CT images. e masses showed an oval shape, irregular margins, and an internal density > 20 HU on unenhanced CT images.
ey were subsequently proven to be malignant by CT-guided core needle biopsy.
Fig 1. A 58-year-old man with known rectal cancer was found to have a 19-mm right adrenal mass (arrow) during a metastatic workup.
e mass had a density of 14 HU on unenhanced CT images, and had smooth margins, an oval shape, and a homogeneous enhancement.
As the mass showed no signicant change at the 20-month CT follow-up, it was considered to be benign.
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REFERENCES
1. Song JH, Chaudhry FS, Mayo–Smith WW. e incidental
adrenal mass on CT: prevalence of adrenal disease in 1,049
consecutive adrenal masses in patients with no known malignancy.
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2. Boland GW, Lee M, Gazelle GS, Halpern EF, McNicholas MM,
Mueller PR. Characterization of adrenal masses using unenhanced
CT: an analysis of the CT literature. AJR Am J Roentgenol.
1998;171:201-4.
3. Lee MJ, Hahn PF, Papanicolaou N, Egglin TK, Saini S, Mueller
PR, et al. Benign and malignant adrenal masses: CT distinction
with attenuation coefficients, size, and observer analysis.
Radiology. 1991;179:415-8.
4. Seo JM, Park BK, Park SY, Kim CK. Characterization of lipid-
poor adrenal adenoma: chemical-shi MRI and washout CT.
AJR Am J Roentgenol. 2014;202:1043-50.
5. Blake MA, Kalra MK, Sweeney AT, Lucey BC, Maher MM,
Sahani DV, et al. Distinguishing benign from malignant adrenal
masses: multi-detector row CT protocol with 10-minute delay.
Radiology. 2005;238:578-585.
6. Sangwaiya MJ, Boland GW, Cronin CG, Blake MA, Halpern
EF, Hahn PF. Incidental adrenal lesions: accuracy of characterization
with contrast-enhanced washout multidetector CT—10-minute
delayed imaging protocol revisited in a large patient cohort.
Radiology. 2010;256(2):504-10.
7. Song JH, Grand DJ, Beland MD, Chang KJ, Machan JT,
Mayo–Smith WW. Morphologic features of 211 adrenal masses
at initial contrast-enhanced CT: can we dierentiate benign
from malignant lesions using imaging features alone? AJR Am
J Roentgenol. 2013;201:1248-53.
8. Hussain S, Belldegrun A, Seltzer SE, Richie JP, Gittes RF,
Abrams HL. Dierentiation of malignant and benign adrenal
masses: predictive indices on computed tomography. AJR Am
J Roentgenol. 1985;144:61-65.
9. Berland LL, Koslin DB, Kenney PJ, Stanley RJ, Lee JY. Dierentiation
between small benign and malignant adrenal masses with
dynamic incremented CT. AJR Am J Roentgenol. 1988;151:95-
101.
10. Ctvrtlík F, Herman M, Student V, Tichá V, Minarík J. Dierential
diagnosis of incidentally detected adrenal masses revealed on
routine abdominal CT. Eur J Radiol. 2009;69(2):243-52.
11. Mayo–Smith WW, Song JH, Boland GL, Francis IR, Israel GM,
Mazzaglia PJ, et al. Management of Incidental Adrenal Masses:
A White Paper of the ACR Incidental Findings Committee. J
Am Coll Radiol. 2017;14(8):1038-44.
12. Na Songkhla N, Chaikittisilpa N, Muangsomboon K. Analysis
of MDCT Findings in the Dierentiation of Adrenal Masses in
Lung Cancer Patients in Siriraj Hospital. Siriraj Med J.
2017;65(2):36-40.
13. Park SH, Kim MJ, Kim JH, Lim JS, Kim KW. Dierentiation of
adrenal adenoma and nonadenoma in unenhanced CT:
new optimal threshold value and the usefulness of size criteria
for dierentiation. Korean J Radiol. 2007;8:328-35.
14. Northcutt BG, Raman SP, Long C, Oshmyansky AR, Siegelman
SS, Fishman EK, et al. MDCT of adrenal masses: Can dual-
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834-39.
Thiravit et al.
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Kusol Russameecharoen, M.D., Katika Nawapun, M.D., Buraya Phattanachindakun, M.D., Vitaya Titapant,
M.D., Tuangsit Wataganara, M.D., Nisarat Phithakwatchara, M.D.
Department of Obstetrics and Gynecology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok 10700, ailand.
Normative Values of Second-Trimester Maternal
Serum Markers Using an Automated Assay Platform
for Down Syndrome Screening
Corresponding author: Nisarat Phithakwatchara
E-mail: nisaratp@gmail.com
Received 14 August 2017 Revised 16 November 2017 Accepted 8 December 2017
ORCID ID: 0000-0002-2517-4432
doi: http://dx.doi.org/10.33192/Smj.2019.04
ABSTRACT
Objective: Automated chemiluminescent immunoassay has several advantages over manual ELISA with comparable
test performance. Few studies have reported the reference values of the second-trimester serum markers maternal
serum alpha-fetoprotein (AFP), human chorionic gonadotropin (hCG), unconjugated estriol (uE3), and inhibin
A (Inh A) by automated immunoassay in Asian population. Accordingly, this study aimed to determine the
median values of second trimester serum markers as a function of gestational age (GA) in ai population using
an automated immunoassay.
Methods: is prospective cross-sectional study of serum markers in healthy singleton second trimester (14-22
weeks) pregnant women was conducted at Siriraj Hospital from September 2012 to April 2015. Maternal serum
AFP, hCG, uE3, and Inh A were analyzed by automated immunoassay. Predicted median values as a function of
GA were calculated from best-t regression equations.
Results: A total of 1,526 women were included. Median values serum markers were constructed from the following
optimal models: AFP (ng/mL) = 99.082 - 14.195 GA + 0.662 GA
2
, r
2
=0.995; hCG (mIU/mL) = 390168.106 - 35
968.397GA + 876.708GA
2
, r
2
=0.972; uE3 (ng/mL) = -3.388 + 0.274 GA, r
2
=0.997; and, Inh-A (pg/mL) = 1206.875 -
114.171 GA + 3.174 GA
2
, r
2
=0.882. Using the same platform analysis and maternal weight adjustment, the reference
values in ai population were shown to be dierent from those of other ethnicities.
Conclusion: Median values of second-trimester serum markers for ai population were determined. Maternal
weight and the use of population-specic normal values have to be taken into account for Down syndrome screening
in the second trimester.
Keywords: Automated immunoassay; Down syndrome; maternal serum screening; quadruple test; reference values;
second trimester (Siriraj Med J 2019;71: 21-24)
INTRODUCTION
Alterations in the serum levels of alpha-fetoprotein
(AFP), human chorionic gonadotropin (hCG), unconjugated
estriol (uE3), and inhibin A (Inh A) during the mid-
trimester (quadruple test), oer a detection rate of 81%
for a 5% false positive rate for prenatal detection of
Down syndrome.
1,2
e quality of the results from risk
calculation is aected by the analytical performance of
the assay used for serum marker determination, the
accurate dating of pregnancy, population-specic median
values of serum analytes, and the reliable relationship
between serum markers and gestational age.
3-6
Well-
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22
established assays oen provide automated quantitation
of serum AFP, hCG, and uE3, but not Inh A. A new
totally automated quantitative assay for these four serum
biomarkers has recently been developed. is automated
immunoassay is dierent from the manual enzyme-
linked immunosorbent assay (ELISA) in terms of sample
treatment, incubations, washes and detection systems,
attributed to the advantage of less labor and fast analysis.
Up to now, only few studies have been reported regarding
the reference values of these four second-trimester serum
markers analyzed by an automated immunoassay among
Asian population.
7,8
Moreover and importantly, no data
derived from an automated immunoassay is available
for ai population. Accordingly, the aim of this study
was to determine the median values of second trimester
serum markers as a function of gestational age (GA) in
ai population using an automated immunoassay.
MATERIALS AND METHODS
Study population
is prospective validation study was carried out
in pregnant women at 14 to 22 weeks’ gestation who
attended antenatal care at the Division of Maternal-Fetal
Medicine, Department of Obstetrics and Gynecology,
Faculty of Medicine Siriraj Hospital from September
2012 to April 2015. e study protocol was approved
by Siriraj Institutional Review Board (Si 413/2012). In
order to generate median values of serum levels for each
analyte, at a signicance of 5%, standard deviation of 32,
margin of error in estimating mean of 5%, and reservation
for data loss of 10%, 160 subjects were required for each
gestational week (total of 1,600 subjects). Inclusion
criteria were women with singleton pregnancy, ai
racial origin, and ≥ 18 years of age. Gestational age was
estimated by either a reliable menstrual history and/
or by ultrasound examination before 13 +6 weeks of
gestation. Exclusion criteria were multiple pregnancies
or prior invasive prenatal diagnostic procedures before
the time of enrollment. e peripheral blood samples
were shipped for analysis to the laboratory no later
than 2 hours aer blood drawing. Serum sample was
transferred at least 500 μL aliquot.
Sample analysis
A new paramagnetic particle chemiluminescent
immunoassay on the Access 2 Immunoassay Systems
(Beckman Coulter, CA, USA) using Beckman Coulter
Access Reagents (cat. No. 33210, 33500, 33570, and
A36097 for AFP, hCG, uE3, and Inh-A, respectively) was
used. Serum levels of these quadruple markers were then
calculated from a stored, multi-point calibration curve.
e lowest detection thresholds of AFP, hCG, uE3, and
Inh A with 95% condence were 0.5 ng/mL, 0.5 mIU/
mL, 0.017 ng/mL, and < 1 pg/mL, respectively. Results
of invasive prenatal genetic testing were obtained from
voluntary self-reporting, with an absence of reported
abnormalities until the time of birth adjudicated to be
euploid.
Statistical analysis
Demographic characteristics are presented as
numbers and percentages for categorical data and as
mean ± standard deviation or median and interquartile
range (IQR) for continuous data, depending on the
distribution. Median values of AFP, hCG, uE3, and Inh
A were calculated for each completed gestational week.
Regression analysis was used to estimate the relationship
of serum markers and gestational age and the optimal
model was then selected to predict median values of
each marker. Patient results were then stratied into
six groups according to maternal weight (< 45, 45-54.9,
55-64.9, 65-74.9, 75-84.9, and ≥ 85 kg). To adjust for
maternal weight, predicted multiples of the median
(MoMs) values of each marker were calculated from the
best t equation using regression analysis. All data were
analysed statistically by using SPSS (IBM SPSS Statistics
for Windows version 18, Microso Corporation; Chicago,
IL, USA) and GraphPad Prism (GraphPad soware for
Windows version 7.00, San Diego, California, USA).
RESULTS
Of the 1,600 women enrolled in this study, 1,526
(95.38%) women with complete data set were selected
for further analysis. Median maternal age, weight, and
body mass index (BMI) at the time of study enrollment
were 27 years (IQR, 23 - 31), 52 kg (IQR, 47 - 59.25),
and 20.82 kg/m
2
(IQR, 18.90 - 23.51), respectively. Only
3% (46/1526) of this study population were obese (BMI
≥ 30 kg/m
2
). Most patients (1,417/1,526, 92.9%) were
aged less than 35 years and 843 patients (55.24%) were
nulliparous. Median gestational age at delivery and
birthweight were 39 weeks (IQR, 38 - 39) and 3,110 grams
(IQR, 2,890 - 3,330), respectively. All patients included
in this study had a naturally conceived pregnancy, no
one reported being a current smoker.
e regression equations for serum markers as a
function of gestational age (GA) in weeks from 14 to 22
weeks of gestation are described as follows:
AFP (ng/mL) = 99.082 – 14.195 GA + 0.662 GA
2
, r
2
= 0.995, P < 0.001
hCG (mIU/mL) = 390168.106 – 35968.397GA + 876.708GA
2
,
r
2
= 0.972, P < 0.001
Russameecharoen et al.
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23
Original Article
SMJ
uE3 (ng/mL) = -3.388 + 0.274 GA, r
2
= 0.997, P < 0.001
Inh-A (pg/mL) = 1206.875 – 114.171 GA + 3.174 GA
2
,
r
2
= 0.882, P = 0.002.
Median values of all serum markers at each gestational
week calculated from the regression equations are shown
in Fig 1. Serum levels of AFP and uE3 signicantly
increased, and serum levels of hCG signicantly decreased
from 14 to 22 gestational weeks. Serum levels of Inh A
continuously dropped until reaching the nadir at 18
weeks of gestation, then continuously elevated.
Signicant inverse relationships between multiples
of the median (MoMs) serum levels of AFP, hCG, uE3,
and Inh A and maternal weight (Wt) (in kilograms) in
each category were observed with the best-t equations
described, as follows:
AFP (MoMs) = 1.972 – 0.024 Wt + 0.00011 Wt
2
, r
2
= 0.994, P < 0.001
hCG (MoMs) = 1.484 – 0.008 Wt, r
2
= 0.921, P = 0.002
uE3 (MoMs) = 1.322 - 0.006 Wt, r
2
= 0.934, P = 0.002
Inh-A (MoMs) = 1.302 – 0.006 Wt, r
2
= 0.998, P < 0.001
Predicted MoM values of all serum markers adjusted
for maternal weight are shown in Table 1.
Fig 1. Median values of serum markers in ai women by gestational age. (a) alpha-fetoprotein (AFP) levels expressed in ng/ml, (b) human
chorionic gonadotropin (hCG) levels expressed in mIU/ml, (c) unconjugated estriol (uE3) levels expressed in ng/ml, (d) inhibin A (Inh A)
levels expressed in pg/ml.
TABLE 1. Predicted median MoM values of serum markers by maternal weight category.
Maternal weight category Number of cases AFP hCG uE3 Inh A
(MoM) (MoM) (MoM) (MoM)
I < 45 kg 199 1.158 1.148 1.070 1.050
II 45 – 54.9 kg 718 1.060 1.092 1.028 1.008
III 55 – 64.9 kg 376 0.950 1.020 0.974 0.954
IV 65 – 74.9 kg 178 0.849 0.940 0.914 0.894
V 75 – 84.9 kg 39 0.769 0.860 0.854 0.834
VI ≥ 85 kg 16 0.710 0.776 0.791 0.771
Abbreviations: AFP: alpha-fetoprotein; hCG: human chorionic gonadotropin; uE3: unconjugated estriol; Inh A: inhibin A; MoM: multiples
of the median; kg: kilogram.
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24
DISCUSSION
In this study, median values of serum levels of AFP,
hCG, uE3, and Inh A in ai women carrying singleton
pregnancy from 14 to 22 weeks of gestation were generated
using a new Access 2 automated chemiluminescence
immunoassay system. Previous studies supported the
highly correlated results of dimeric Inh A between this
new immunoassay platform and manual ELISA, with
a comparable performance of prenatal detection of
Down syndrome.
9,10
Several advantages of an automated
chemiluminescent immunoassay over a manual ELISA
with a comparable test performance explain its preference
for serum Inh A analysis.
9,10
e eects of gestational age
and maternal weight were consistent with previously
published data from other platforms.
11,12
Maternal weight is another inuential determinant of
these serum marker levels, displaying negative aliations.
e pathophysiology behind these aliations is at present
indistinct. In order to compare the median values of
maternal serum markers in our population to those
previously reported from other populations using the
same automated immunoassay, it is essential to adjust the
median values by weight-correction models. Each of four
serum markers had a similar pattern of change during
the second trimester of pregnancy among various ethnic
groups.
7,9,10,13
Nevertheless, unique normal values were
reported among dierent ethnic groups. Higher levels of
serum AFP, hCG, and Inh A and a lower level of serum
uE3 in our population were observed in comparison
of those in the Caucasian groups.
9,10,13
Despite the fact
that ai and Korean people are both racially classied
as Asian and have similar serum AFP and hCG levels
in the second trimester, there are some more subtle
dierences in uE3 and Inh A levels between these two
ethnic groups.
7
Our population seemed to have a lower
level of serum uE3 and a slower progression of serum Inh
A aer reaching its nadir level at 18 weeks of gestation.
7
ese would seem to signify the necessity of population-
specic normal values of these serum markers.
e potential impact of this study is reinforced by
a number of key strengths. is is the rst prospective,
well-designed study of normal values of second-trimester
serum markers using an automated immunoassay in a
large cohort of ai population with a high rate of available
outcomes. Limitations of this study include a highly selected,
low-risk study population, no accuracy assessment of these
reference values, and no data comparison between the
normal values derived from the automated immunoassay
and those derived from the manual ELISA method in
the studied ai population. e impact of maternal
age, smoking status, and the method of conception on
these reference values could not be determined. Further
studies on test performance are needed to support the
application of these normative values to second-trimester
screening for Down syndrome.
ACKNOWLEDGMENTS
is work was funded by Beckman Coulter Singapore
Pte. Ltd. and PCL Holding Co., Ltd. e study sponsors
supplied the reagents for use in this study. e study sponsors
had no role in the study design, data collection, statistical
analysis and interpretation, manuscript preparation, or
publication decision.
REFERENCES
1. Malone FD, Canick JA, Ball RH, Nyberg DA, Comstock CH,
Bukowski R, et al. First-trimester or second-trimester screening,
or both, for Down's syndrome. N Engl J Med. 2005;353(19):2001-11.
2. Wald NJ, Rodeck C, Hackshaw AK, Walters J, Chitty L, Mackinson
AM, et al. First and second trimester antenatal screening for
Down's syndrome: the results of the Serum, Urine and Ultrasound
Screening Study (SURUSS). Health Technol Assess. 2003;7(11):1-77.
3. MacRae AR, Gardner HA, Allen LC, Tokmakejian S, Lepage
N. Outcome validation of the Beckman Coulter access analyzer
in a second-trimester Down syndrome serum screening
application. Clin Chem. 2003;49(1):69-76.
4. Wald NJ, Hackshaw AK, George LM. Assay precision of serum
alpha fetoprotein in antenatal screening for neural tube defects
and Down's syndrome. J Med Screen. 2000;7(2):74-7.
5. Bishop J, Dunstan FD, Nix BJ, Reynolds TM. e eects of
gestation dating on the calculation of patient specic risks
in Down's syndrome screening. Ann Clin Biochem. 1995;32
(Pt 5):464-77.
6. Reynolds T, Ellis A, Jones R. Down's syndrome risk estimates
demonstrate considerable heterogeneity despite homogeneity
of input. Ann Clin Biochem. 2004;41(Pt 6):464-8.
7. Lee JH, Park Y, Suh B, Song SM, Kwon OH, Kim JH. Performance
characteristics of the UniCel DxI 800 immunoassay for the
maternal serum quadruple test, including median values
for each week of gestation, in Korean women. Korean J Lab
Med. 2010;30(2):126-32.
8. Kwon JY, Park IY, Park YG, Lee Y, Lee G, Shin JC. Korean-specic
parameter models for calculating the risk of Down syndrome in
the second trimester of pregnancy. J Korean Med Sci.
2011;26(12):1619-24.
9. Lambert-Messerlian GM, Palomaki GE, Canick JA. Inhibin
A measurement using an automated assay platform. Prenat
Diagn. 2008;28(5):399-403.
10. Rawlins ML, La'ulu SL, Erickson JA, Roberts WL. Performance
characteristics of the Access Inhibin A assay. Clin Chim Acta.
2008;397(1-2):32-5.
11. Wanapirak C, Sirichotiyakul S, Luewan S, Yanase Y, Traisrisilp
K, Tongsong T. Dierent median levels of serum triple markers
in the second trimester of pregnancy in a ai Ethnic Group.
J Obstet Gynaecol Res. 2012;38(4):686-91.
12. Promsonthi P, Panburana P, Kadegasem P, Chaemsaithong P,
Preechapornprasert D, Chanrachakul B. Inhibin-A levels
between 14 and 20 weeks of gestation in ai women. J Obstet
Gynaecol Res. 2012;38(1):118-21.
13. Vranken G, Reynolds T, Van Nueten J. Medians for second-
trimester maternal serum markers: geographical dierences
and variation caused by median multiples-of-median equations.
J Clin Pathol. 2006;59(6):639-44.
Russameecharoen et al.
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25
Original Article
SMJ
Prapaporn Noparatayaporn, M.Sc.*, **, Tanita aweethamcharoen, Ph.D.*, **, Apirom Laocharoenkeat, M.S.**,
Panitta Narkchuay, M.N.S. ***, Anchalika Klinniyom, M.Sc. (in Pharm)**, Cherdchai Nopmaneejumruslers,
M.D.****, Khemchat Wangtawesap, M.D. *****, Siriporn Pitimana-aree, M.D. *****, Darin Sakiyalak, M.D.******
*Siriraj Health Policy Unit, **Department of Pharmacy, ***Department of Nursing, **** Department of Medicine, *****Department of Anesthesiology,
******Department of Ophthalmology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok 10700, ailand.
Economic Evaluation of Ready-to-use Injectable
Medications by Pharmacy Department Compared
with the Traditional System of Individual Preparation
by Nurse
Corresponding author: Tanita aweethamcharoen
E-mail: tanitath@gmail.com
Received 24 May 2017 Revised 27 October 2017 Accepted 8 December 2017
ORCID ID: 0000-0002-6093-779X
http://dx.doi.org/10.33192/Smj.2019.05
ABSTRACT
Objective: To dispense medication in a form of ready to use (RTU) medication was recommended by the standards of
Joint Commission International (JCI) and Standard Guidelines of Hospital Pharmacy for preventing the medication
error. However, the cost and benet were questionable. e costs may increase while benets were unclear. Before
making the implementation decision, the cost of investment and benet between traditional (injectable medication
is prepared by nurse) and RTU systems (injectable medication is prepared by pharmacy department) should be
evaluated.
Methods: is study compared the cost and benet of injectable medication administration between the traditional
system and the RTU system within a large academic hospital. e decision tree was designed to produce comparable
data on the hospital’s perspective. e time horizon was 10 years thus all costs were discounted at 3% annually.
Sensitivity analysis was performed to test the stability of the results.
Results: e cost of investment at 10-year intervals of the RTU system was lower than the traditional system by
about 18,710,160 baht. e benet was decreased 19.32 full time equivalents (FTEs) of nurse when compared with
the traditional system. e result showed that the ve most sensitive variables were number of doses, mixing time
per dose (prepared by nurse), space for production, salary and fringe benets of pharmacists, and mixing time per
dose (prepared by pharmacist).
Conclusion: e RTU system saved 1,871,016 baht per year and 19.32 FTEs of nurse. Moreover, the RTU system
enhanced the opportunity of nurses and pharmacists to play more professional role and promoted the ecient
health care system.
Keywords: Ready-to-use medication; premixed medication; intravenous admixture; medication administration;
economics (Siriraj Med J 2019;71: 25-30)
INTRODUCTION
The growing nursing workforce shortage has
increased nurse workload and reduced the amount of
nursing time available for patient care activities.
1
e prior
study found that nurses time spend on specic activities
such as documentation, medication administration,
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26
care coordination, and patient care activities.
2
In the
hospital view, some parts of nurses’ activity especially
admixing in step of medication administration would
be considered to decrease workload of nurse. Some
injection medications such as high alert drugs were
prepared by pharmacy department and dispensed in
ready to use dosage form. us medication would be
administered to the patient without the admixing step
by nurse. Totally, nurses could decrease their workload
and have more times for patient care. From previous
study, 73.3% of nurses agreed that ready to use (RTU)
medication preparation reduced nurses’ workload.
3
For the pharmacist’s role, dispensing medications in
form of ready-to-use medications or premixed medications
in order to decrease medication errors was recommended
by the standard of Joint Commission International (JCI)
and Standard Guidelines of Hospital Pharmacy.
4
Having
RTU medications may help reduce admixing errors or
administration errors.
5
us, to prevent the medication
error, enhance pharmacist role, and decrease nurse
workload, RTU medication was considered. Previous
study found that advantages of RTU medications are
as follows:
1. RTU medications assured that patients received
accurate dosages, and reduced medication errors.
6-10
2. RTU medications could be administered more
quickly especially in a busy time in hospital. us they
could reduce overloading in emergency rooms and other
treatment areas.
10
3. e cost of RTU medications preparation was
less than cost of individual preparation by nurse.
10,11
4. RTU medications reduced risk of microbial
contamination and cross contamination.
10
5. RTU medications could reduce needle-prick
injury which was a major occupational health and safety
issue facing healthcare professionals especially nurses.
10
Previous studies
6-9
found that injectable medication
preparation by pharmacist decreased medication errors
whereas the system may increase cost of investment.
However, the overall cost was expected to be decreased
from prevented medication errors.
For this setting, the cost of investment for the RTU
system and the evidence of benet were controversial
issues that provoke diering views from the relevant
health care personnel. e costs may increase from the
capital cost for the standard practice of sterile preparation
and the involved personnel in the production process
while benets were unclear. Before RTU medication
implementation, cost of investment and benet of the
system needs to be evaluated. us the objective of the
study was comparison on the cost of investment and
benet of injectable medication preparation system
between traditional and RTU systems.
MATERIALS AND METHODS
e study evaluated two intravenous admixture
systems; the traditional system (injectable medications
were prepared by nurse) and the RTU system (injectable
medications were prepared by pharmacy department.
Nine high volume antibiotics aer reconstitution is stable
under refrigeration (2-8 °C) for at least seven days were
chosen for RTU medication. e recommended dose
of preparation was the usual adult dose for treatment in
the hospital. Cost of investment was analyzed in hospital
perspective. Direct cost including capital cost, labor
cost, material cost, and other costs incurred in ten years
were accumulated and discounted to present value with
3% discounting rate.
12
To analyze cost and sensitivity
analysis, TreeAge Pro Healthcare was used. e benet
in terms of full-time equivalents (FTE), of the number
of sta required for work process, was analyzed for
comparison. In medication preparation process, the
traditional system required nurse, whereas pharmacist
and pharmacy technician were involved in the RTU
system. e FTE of the study was calculated based on
6 hours per day and 230 days per year. Data included
in the study was extracted from the hospital data and
directly recorded at ward and pharmacy department.
(is study was approved by the Institutional Review
Board Ethics Committee IRB. No. 558/2558(EC2).)
Traditional system
Injectable medication was prepared for administration
with aseptic technique at ward environment by nurse.
en medication would be immediately administered
to patient. Cost of investment comprised of labor cost
and material cost. Labor cost including salary and fringe
benets of nurse was calculated from time spent in
preparation process. Time spent of work process was
collected at ward and calculated as full-time equivalent
(FTE). Data of salary and fringe benets were obtained
from human resource unit and estimated that in every
year salaries increased 5% (salary increase between 3-7%
per year). Sterile syringe and needle were used and
accumulated for cost per dose in preparation process.
RTU system
Injection medication was prepared by pharmacist
(R.Ph) and pharmacy technician (Ph.Tech) in cleanroom
as the standard practice of pharmaceutical compounding
- sterile preparations. RTU medication was stored at
controlled temperature (2-8 °C) and dispensed with cold
Noparatayaporn et al.
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Original Article
SMJ
chain system. Cost of investment consisted of capital
cost, labor cost, material cost, and other costs. Capital
costs were xed cost incurred in the production of RTU
medication such as laminar airow hood, vial roller mixer,
sealer, repeater, pharmaceutical refrigerator, autoclave,
hot air oven. For this study, a lifetime of 10 years has
been chosen for all production equipment by an expert
panel. Computer and printer would be changed every
3 - 5 years. Cleanroom and laminar airow hood needed
maintenance since the rst year of production. For other
equipment, for the rst two years of implementation
the support and maintenance were free. In this system,
opportunity cost of space for production was included.
Labor cost was dened as labor cost of pharmacist and
pharmacy technician. Time spent in preparation and
packaging process was collected at pharmacy department.
Data of salary and fringe benets were obtained from
human resource unit and estimated that in every year
salaries increased 5%. RTU medication was contained
within the sterile packaging labeled medication name,
concentration, lot number, and expiration date. Material
including sterile syringe, sterile needle, packaging, and
labeling was used for RTU medication.
Sensitivity analysis
Sensitivity analysis was used to evaluate how
uncertainty in the model inputs aected the outputs
of the model. ere was a wide range of input data for
sensitivity analysis as Table 1. e use of sensitivity
analysis results were classied to four categories: making
decision or development of recommendations for decision
makers, communication, increased understanding or
quantication of the system, and model development.
While all these uses were potentially important, this
study focused on decisions making or recommendations.
When the optimal option was insensitive to parameter
changes, policy maker considered higher condence of
implementing the optimal option. On the other hand,
if the option was sensitive, sensitivity analysis could be
used to specify the level of importance of changes and
recommend solutions. Even if the levels of variables
in the optimal solution were changed dramatically by
a higher or lower parameter value, the stability of the
outcomes should examine the dierence in prot (or
another relevant objective) between these solutions and
the base-case solution.
13
One way sensitivity analysis was
performed on all variables in order to test the stability
of the outcomes and presented as the tornado diagram.
Tornado diagram showed changes in the net present
value under the feasible range of each variable.
RESULTS
e estimated number of medications per year was
300,827 doses. 10-year cost of investment for traditional
and RTU system were analyzed and presented as present
value. Direct cost was accumulated from capital cost,
labor cost, material cost, and other costs. e total 10-
year cost of the traditional system included labor and
material costs which was 98.24 and 9.25 million baht,
respectively. For the RTU system, total 10-year cost of
labor and material were 39.52 and 5.07 million baht,
respectively. Capital and opportunity costs were also
included in the RTU system. Total 10-year capital and
opportunity costs were 4.65 and 39.54 million baht,
respectively. e cost of investment of the traditional
system and the RTU system were 107,492,820 baht and
88,782,660 baht, respectively as shown in Table 2. e
RTU system reduced overall cost about 18,710,160 baht
in 10 years when compared with the traditional system.
For workload, the RTU system could shorten some
preparation processes so lower human resource requirements
per dose were needed. As Table 1, medication preparation
time in the traditional system took 319 seconds per dose
compared with 240 seconds per dose in the RTU system.
e result showed that the traditional system required
19.32 FTEs of nurses while the RTU system required
3.63 FTEs of pharmacists and 10.90 FTEs of pharmacy
technicians. us using the RTU system could replace
19.32 FTEs of nurses.
Sensitivity analysis
From the Fig 1, the most sensitive variable was
number of doses per year of RTU medication. e RTU
system saved cost when the minimum number of RTU
medications was 211,346 doses per year. e following
sensitive variables were nursing time for medication
preparation, space of production, salary and fringe benets
of pharmacist, and pharmacy technician, respectively.
Decreasing the nursing time for medication preparation
from 319 sec/dose to 258 sec/dose, the lowest expected
value changed from the RTU system to the traditional
system. When the space of production was more than
552.46 square meters, the lowest expected value changed
from the RTU system to the traditional system. Increasing
the salary and fringe benets per month of pharmacist
and pharmacy technician to 77,018.29 and 28,191.26
baht, respectively, the lowest expected value changed
from the RTU system to the traditional system. For the
salary and fringe benets of nurse with less than 31,418.62
baht/month, the lowest expected value changed from the
RTU system to the traditional system. With the increase
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28
TABLE 1. All variables and intervals used in the model and sensitivity analysis.
Variable Value Low High
Number of medications (doses/year) 300,827 220,934 391,075
Working time per year (sec/FTE) 4,968,000 4,968,000 5,796,000
Increasing rate of salary per year (%) 5 3 7
Discount rate (%) 3 0 7
Traditional system
Medication preparation time: nurse (sec/dose) 319 30 2,730
Salary and fringe benets of nurse (baht/month) 38,809.96 23,250.00 124,257.10
Material cost (baht/dose) 3.50 2.50 5.00
Ready to use (RTU) system
Laminar Airow Hood cost (baht/piece) 400,000 300,000 500,000
Vial Roller Mixer cost (baht/piece) 25,000 25,000 30,000
Sealer cost (baht/piece) 25,000 25,000 30,000
Repeater 300,000 300,000 350,000
Pharmaceutical Refrigerator cost (baht/piece) 38,000 26,000 38,000
Autoclave cost (baht/piece) 350,000 350,000 400,000
Hot Air Oven cost (baht/piece) 130,000 130,000 150,000
Computer and Software cost (baht/piece) 25,000 20,000 30,000
Printer cost (baht/piece) 20,000 20,000 25,000
Refrigerator for storing RTU medications 128,000 104,000 152,000
Maintenance cost of Laminar Airow Hood (baht/machine/year) * 7,000 4,900 9,100
Maintenance cost of cleanroom (baht/year) * 60,000 60,000 100,000
Maintenance cost of product equipment (baht/year)
†
50,000 35,000 65,000
Space of production (square metre : Sq m) 375 255 555
Opportunity cost (baht/m
2
/year) 12,000 12,000 18,000
Medication preparation time: R.Ph (sec/dose) 60 36 120
Medication preparation time: Ph.Tech (sec/dose) 180 120 180
Salary and fringe benets of R.Ph (baht/month) 37,720.96 21,290.00 124,236.90
Salary and fringe benets of Ph.Tech (baht/month) 15,092.14 11,422.00 54,291.30
Material cost (baht/dose) 1.92 1.51 9.00
Abbreviations: R.Ph = Pharmacist, Ph.Tech = Pharmacy technician
* Cleanroom and laminar airow hood required maintenance every 6 months since the rst year of implementation.
†
Maintenance cost occurred in the third year aer implementation.
Noparatayaporn et al.
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Original Article
SMJ
Fig 1. Tornado diagram
in material cost for RTU preparation to 9 baht/dose,
the traditional system would be cost saving. Not only
space of production, but opportunity cost of space also
aected the alternative. Increasing the opportunity cost
from 1,000 baht/m
2
/month to 1,473 baht/m
2
/month, the
lowest expected value changed from the RTU system to
the traditional system.
DISCUSSION
is study focused on medication in standard dose
with extended shelf life at least seven days. e estimated
numbers of medications per year were 300,827 doses. In
the RTU system, capital, maintenance, and opportunity
costs increased. However, the RTU system saved labor
and material costs. Material cost per dose in the RTU
system lowered from fewer needle and syringe volumes
per dose preparation. As a result, cost saving of labor and
material costs from the RTU system were 58,725,439 baht
and 4,176,096 baht in 10 years, respectively. Overall in a
10-year period, the RTU system saved 18,710,160 baht.
As a result, the RTU system could save nursing time 19.32
FTEs from reducing administration time while workload
of pharmacist and pharmacy technician increased 3.63
FTEs and 10.90 FTEs, respectively. e benets of the
RTU system implementation were the opportunity of
nurse to provide a patient care and pharmacists to play
the role as the standard of JCI and good pharmacy
practice. Moreover, the RTU system is able to enhance
the safety of medication administration and promote
the ecient healthcare system.
From sensitivity analysis, the number of medications
was the most sensitive variable. To increase items covered
by the RTU system could save more cost of the system.
e result would be inuenced by the change of number
of medications, admixing time per dose by nurse, space
for production and opportunity cost, salary and fringe
benets of pharmacist, pharmacy technician, and nurse,
and material cost of the RTU system. To decrease labor
cost in the RTU system, oering overtime compensation
could decrease labor cost compared with full-time sta
employment. When nurse took admixing time less than
258 seconds, the traditional system would be the lower
cost alternative. In this observation, 44.43% of admixing
doses took less than 258 seconds. However, nursed can
be disturbed by calls and other notications during
administer or mixing injectable drug. e previous study
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30
showed that the interruption event occurred one in 1.7
times of admixing. us nurse required more time in
real practice during admixing from the interrupted event.
e study did not take into account the cost of
medication and diluent which were equal in both systems.
RTU medication was the dose of standard usually used
with long stability, so cost from expired RTU medications
was not accumulated. However, the ecient system
would be planned for the RTU system implementation
to prevent waste. Cost saving from unused medication
in vial as prior study was not included because dose of
RTU medication was similar to the company packaging.
3
Initially, the RTU system was implemented in nine
antibiotics and cost saving from reducing of administrative
errors was not accumulated as study from Colombia
which explored in high alert medication which cause
patient harm.
14
CONCLUSION
e RTU system was the lowest cost alternative, saves
time and workload of nurse by eliminating reconstitution
at the point of care. As well as the economic interest, the
RTU system could enhance the opportunity of nurses
and pharmacists to play more professional role and
contribute to patient safety improvement and hospital
quality following the standard of JCI.
ACKNOWLEDGMENTS
This research was successfully achieved by the
cooperation of sta in nursing, pharmacy department,
and Siriraj Health Policy unit during the period of data
collection, which was information from their routine
job. is research was supported by grant from the
Routine to Research (R2R) of Siriraj Hospital, Mahidol
University, ailand.
REFERENCES
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Francisco: Health Workforce Solutions LLC; 2007.
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Time and Motion Study: How Do Medical-Surgical Nurses
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4. (ailand) TAoHP. Standard Guidelines of Hospital Pharmacy
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accuracy in compounding i.v. admixtures at ve hospitals.
Am J Health Syst Pharm.1997;54(8):904-12.
6. Webster CS, Merry AF, Gander PH, Mann NK. A prospective,
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7. ASHP Expert Panel on Medication Cost Management. ASHP
guidelines on medication cost management strategies for hospitals
and health systems. Am JHealthSyst Pharm.2008;65(14):
1368-84.
8. Crawford SY, Narducci WA, Augustine SC. National survey of
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413.
9. Salberg DJ, Newton RW, Leduc DT. Cost of wastage in a hospital
intravenous admixture program. Hosp Formul.1984;19(5):
375-8.
10. Makwana S, Basu B, Makasana Y, Dharamsi A. Prefilled
syringes: An innovation in parenteral packaging. Int J Pharm
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11. Armoiry X, Aubrun F, Aulagner G, Piriou V. Economic impact
of ephedrine prelled syringes in anaesthesiology. Ann Fr
Anesth Reanim.2014;33(1):47-8. [Article in French]
12. Drummond MF, Sculpher MJ, Torrance GW, O’Brien BJ,
Stoddart GL. Methods for the economic evaluation of health
care programmes third edition. 2nd ed. Oxford: Oxford University
Press, 2005.
13. Pannell DJ. Sensitivity analysis of normative economic models:
eoretical framework and practical strategies. Agricultural
Economics 1997;16:139-52.
14. Rosselli D, Rueda JD, Silva MD, Salcedo J. Economic Evaluation
of Four Drug Administration Systems in Intensive Care Units
in Colombia. Value Health Reg Issues.2014;5:20-24.
Noparatayaporn et al.
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31
Original Article
SMJ
Saran Maneesuwansin, M.D., Chalairat Suk-ouichai, M.D., Patkawat Ramart, M.D., Siros Jitpraphai, M.D.,
Kittipong Phinthusophon, M.D., Ekkarin Chotikawanich, M.D., Teerapon Amornvesukit, M.D., Tawatchai
Taweemonkongsap, M.D., Bansithi Chaiyaprasithi, M.D., Sunai Leewansangtong, M.D., Sittiporn Srinualnad,
M.D., Chaiyong Nualyong, M.D.
Department of Surgery, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok 10700, ailand.
Administration of Renin-Angiotensin System
Inhibitor Affects Tumor Recurrence and Progression
in Non-Muscle Invasive Bladder Cancer Patients
Corresponding author: Chalairat Suk-ouichai
Email: chalairat.suk@mahidol.ac.th
Received 6 February 2018 Revised 15 March 2018 Accepted 21 March 2018
ORCID ID: 0000-0003-3175-0886
http://dx.doi.org/10.33192/Smj.2019.06
ABSTRACT
Objective: To evaluate the eects of renin-angiotensin system inhibitors (RASIs) on tumor-recurrence and disease-
progression in non-muscle invasive bladder cancer (NMIBC) patients.
Methods: From 2006-2015, 348 NMIBC patients at Siriraj Hospital were recruited for this study. Tumor-recurrence
was identied aer the transurethral resection of bladder cancer (TUR-BT) and pathological conrmation of NMIBC,
while stage-progression was dened as muscularis-propria invasion aer pathological review or metastases. Cox
proportional hazards models were used to assess the recurrence-free survival (RFS) and progression-free survival
(PFS) rates.
Results: Of the 348 patients, 86 (24.7%) received RASIs at the rst TUR-BT. e median age was 68 years, and it
was signicantly older for the RASI cohort. No dierences in the tumor characteristics of the groups were found.
e median follow-up periods for tumor-recurrence and stage-progression were 2.3 and 3.7 years, respectively.
Forty percent of the patients experienced tumor-recurrence, with the no-RASI cohort experiencing a signicantly
higher tumor-recurrence rate (46% versus 22%, p<0.001). e 5-year RFS rates were 54% and 78% for the no-RASI
and RASI cohorts, respectively (p=0.001). Stage-progression was observed in 6% of the patients. e 5-year PFS
rates were 87% and 97% for the no-RASI and RASI cohorts, respectively. On univariate and multivariate analyses,
a tumor size ≥3 cm and tumor multifocality were associated with recurrent bladder cancer (p<0.02). On the other
hand, the administration of RASIs was associated with a reduced recurrence (p≤0.002).
Conclusion: Our study suggests that RASI administration might be a potential factor to prevent bladder cancer
recurrence. Further study is needed to evaluate the eects of RASIs.
Keywords: Non-muscle invasive bladder cancer; renin-angiotensin system inhibitors; tumor recurrence; stage
progression (Siriraj Med J 2019;71: 31-37)
Abbreviations
AJCC = American Joint Committee on Cancer
Ang II = Angiotensin II
AT1R = Angiotensin type 1 receptor
BCG = Bacillus Calmette–Guerin
CI = Condence interval
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EORTC = European Organization for Research and Treatment of Cancer
HR = Hazard ratio
IQR = Interquartile range
MIBC = Muscle invasive bladder cancer
MMC = Mitomycin C
MP = Muscularis propria
NMIBC = Non-muscle invasive bladder cancer
OR = Odd ratio
PFS = Progression-free survival
RASI = Renin-angiotensin system inhibitor
RFS = Recurrence-free survival
RR = Relative risk
TUR-BT = Transurethral resection of bladder tumor
INTRODUCTION
Bladder cancer is the second most common urologic
malignancy and the eighth most common overall malignancy
in ailand.
1
Most patients present with gross hematuria,
and 75% of patients have non-muscle invasive bladder
cancer (NMIBC) stages Ta, T1, and Tis.
2-4
Transurethral
resection of bladder tumor (TUR-BT) is the standard
treatment and the diagnostic procedure.
2-4
Up to 50%
of NMIBC patients experience tumor recurrence, and
6% -17% progress to muscle invasive bladder cancer
(MIBC).
4
e use of adjuvant agents aer a TUR-BT has
been introduced to reduce the risk of tumor recurrence
and progression.
2-4
Adjuvant intravesical therapy, such as Bacillus
Calmette–Guerin (BCG) and mitomycin C (MMC),
has been utilized in current practice to decrease the
incidences of recurrent NMIBCs and disease progression.
2-6
A recent meta-analysis demonstrated that adjuvant
intravesical BCG as immunotherapy was associated with
reduced recurrent NMIBC (RR 0.56, 95% CI 0.43-0.71).
5
Additionally, intravesical chemotherapy such as MMC,
doxorubicin, and epirubicin have also been associated
with a decreased risk of bladder cancer recurrence (RR
0.68, 95% CI 0.55 - 0.83; RR 0.80, 95% CI 0.72 - 0.88;
and RR 0.63, 95% CI 0.53 - 0.75, respectively). As to
tumor progression, only adjuvant intravesical BCG
has been associated with a reduced risk of progression
(RR 0.39, 95% CI 0.24 - 0.64). Given its potential role
in both reduced tumor recurrence and progression,
immunotherapy such as checkpoint blockade has also
been studied as an option for the treatment of NMIBC
patients.
7
However, all agents are adjuvant treatments
aer the TUR-BT.
Angiotensin II (Ang II) is a key biological peptide
in renin-angiotensin systems. Ang II is involved in the
regulation of blood pressure, water, and sodium homeostasis,
and in the control of other neurohumoral systems. It
also leads to the excessive production of reactive oxygen
species, and to the hypertrophy, proliferation, migration,
and apoptosis of vascular cells.
8
Angiotensin type 1
receptors (AT1Rs) are expressed in various malignancies,
including bladder cancer, and are signicantly involved
in tumor growth, metastasis, and angiogenesis.
9
Ang II-
AT1R signaling leads to the potent induction of vascular
endothelial growth factors.
10
Recent publications have
outlined that renin-angiotensin system inhibitors (RASIs)
have an antiangiogenic eect on bladder cancer.
11–12
Our primary objective was to determine the eects
of RASIs on tumor recurrence and disease progression
in NMIBC patients.
MATERIALS AND METHODS
Aer receiving Siriraj Institutional Review Board
approval (Si 708/2015), patients diagnosed with NMIBC
at Siriraj Hospital between 2006 and 2015 were recruited
for the study. Excluded were those patients who were
followed up for less than 1 year, underwent a cystectomy
due to unresectable lesions, took RASIs aer the initial
TUR-BT, or had concurrent upper urinary tract tumors.
A total of 348 patients were ultimately available for the
study.
e patient and tumor characteristics were collected
retrospectively. e RASIs had been prescribed by physicians
as anti-hypertensive drugs or for other indications, such
as cardiac or renal disorders. e pathological reports
were in accord with the guidelines of the American Joint
Committee on Cancer (AJCC) current during that period.
NMIBCs were dened as Ta, TI or Tis. Regarding the
past records, some tumors were reported pathologically
as NMIBC by non-muscularis propria invasion (Tu).
MIBCs had been conrmed by histological muscularis
propria invasion (T2-T4). Tumor-recurrence had been
identied aer the TUR-BT and pathological conrmation
of NMIBC. Stage-progression was identied as MIBC or
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lymph nodes or distant metastases. Tumor multifocality
was dened as the presence of 2 or more tumors. e
surveillance schedules had followed the standard guidelines
for each NMIBC risk group. e TUR-BT technique
and the administration of intravesical therapy depended
upon the preferences of the attending sta.
Continuous variables were shown as medians and
interquartile ranges (IQRs), and they were compared
with the Mann–Whitney U test. Categorical variables
were presented as numbers (percent) and compared with
the Chi-square or Fisher’s exact tests. Cox proportional
hazards models were employed to assess the recurrence-free
survival (RFS) and progression-free survival (PFS) rates.
e RFS curve was generated using the Kaplan-Meier
method and compared with the log rank test. Variables
with p<0.05 were considered signicant. e analyses
were performed using SPSS Statistics for Windows,
version 17.0 (SPSS Inc., Chicago, IL, USA).
RESULTS
Overall, 348 patients were analyzed. eir median
age was 68 years. e patient and tumor characteristics
are listed in Table 1. Of those patients, 86 (25%) were
taking RASIs at the time of the rst TUR-BT. e types of
RASI are listed in Table 2. Male gender was predominant
in both cohorts. e smoking histories, tumor sizes,
grades, and multifocality of the cohorts were similar.
One hundred and eighty-one patients (29%) had never
smoked, 101 (52%) used to smoke, and 66 (19%) were
current smokers. e tumor size was less than 3 cm in
233 patients (67%), while it was equal to, or greater than,
3 cm in 115 patients (33%). Ta was found in 95 patients
(27.3%), T1 and Tis in 43 patients (12.4%), and Tu in
210 patients (60.3%). One hundred and sixty-eights
patients (48%) had high grade tumors, with 119 and 49
patients in the no-RASI and RASI cohorts, respectively.
Multifocal tumors were found in 170 patients (49%), with
130 and 40 patients in the no-RASI and RASI cohorts,
respectively. MMC was administrated aer the TUR-
BT in 50 patients (14.4%), while 113 patients (32.5%)
received BCG therapy, and a further 25 (7.2%) received
both agents.
e median follow-up times for tumor recurrence
and stage progression were 2.3 years (IQR 1.1-4.2) and
3.7 years (IQR 2.0-5.8), respectively. One hundred and
forty patients (40%) experienced tumor recurrence, with
patients in the RASI cohort having a signicantly lower
rate of tumor-recurrence (22% versus 46%, p<0.001), as
shown in Table 3. e 5-year recurrence-free survival
(RFS) rates were 54% and 78% for the no-RASI and RASI
cohorts, respectively (p=0.001). e RFS between the 2
cohorts was demonstrated with Kaplan–Meier curves (Fig
1). Stage progression was observed in 19 patients (6%),
with 3 and 16 patients in the RASI and no-RASI cohorts,
respectively (p=0.5). e 5-year PFS rates were 87% and
97% for the no-RASI and RASI cohorts, respectively.
On univariate and multivariate analyses (Table
4), a tumor size equal to or greater than 3 cm, tumor
multifocality, and patients without RASIs were associated
with recurrent bladder cancer (all p<0.02). However, there
was no signicant association between RASI administration
and decreased disease progression in both the univariate
and multivariate analyses.
DISCUSSION
In ailand, bladder cancer is the second most common
urologic malignancy and the eighth most common overall
malignancy, with a prevalence of 4.5/100,000 in males and
of 1.2/100,000 in females.
1
NMIBC is the most common
presentation, and a variety of adjuvant treatments have
been investigated to prevent tumor recurrence and disease
progression.
2-7
In a recent meta-analysis, only intravesical
BCG demonstrated an association with reduced bladder
cancer recurrence and progression in NMIBC patients.
5
In
contrast, adjuvant chemotherapy has been solely associated
with decreased tumor recurrence, not progression.
5
However, the adverse eects of intravesical BCG are still a
concern. Targeted therapies such as checkpoint inhibitors,
which function as immunotherapy, have been explored
for their substantial eects.
7
In addition, the AT1R has
been found in various cancer cells and been shown to be
involved in tumor growth and angiogenesis.
9-10
Shirotake
and colleagues, who studied the AT1R of bladder cancer
patients, reported that it was an independent predictor
of the RFS rate on multivariate analysis.
9
As such, RASIs,
which are prescribed as anti-hypertensive drugs, may
have a potential role in diminishing the risks of tumor
recurrence and progression.
e AT1R has been found in bladder cancer specimens
and has been signicantly associated with intramural
neovascularization.
9
AT1R could therefore be a potential
factor to identify patients with a high risk of tumor
recurrence. Ang II-AT1R also has an impact on tumor
microenvironments and thus promotes tumor growth,
survival, invasive behavior, and tumor cell migration.
10
is suggests that RASIs, such as angiotensin-converting
enzyme inhibitors and angiotensin receptor blockages,
which are prescribed as anti-hypertensives, might have
substantial roles. Blocking the AT1R might reduce tumor
growth and angiogenesis and, in turn, inhibit tumor
proliferation.
9,10
Previous studies from Yuge et al. and
Blute et al. showed that the administration of RASIs was
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TABLE 1. Patient and tumor characteristics.
All patients No-RASIs RASIs P-value
(n=348) (n = 262) (n = 86)
Age , years, median (IQR) 68.2 (30.9-93.2) 0.03
< 65 years, n (%) 127 (36.5) 104 (39.7) 23 (26.7)
≥ 65 years, n (%) 221 (63.5) 158 (60.3) 63 (73.3)
Gender, n (%) 0.89
Male 261 (75.0) 196 (74.8) 65 (75.6)
Female 87 (25.0) 66 (25.2) 21 (24.4)
Smoking, n (%) 0.10
Never 181 (29.0) 135 (51.5) 46 (53.5)
Former 101 (52.0) 71 (27.1) 30 (34.9)
Current 66 (19.0) 56 (21.4) 10 (11.6)
Tumor size, n (%) 0.37
< 3 cm 233 (67.0) 172 (65.6) 61 (70.9)
≥ 3 cm 115 (33.0) 90 (34.4) 25 (29.1)
Tumor stage, n (%) 0.04
Ta 95 (27.3) 68 (26.0) 27 (31.4)
T1+Tis 43 (12.4) 27 (10.3) 16 (18.6)
Tu 210 (60.3) 167 (63.7) 43 (50.0)
Tumor grade, n (%) 0.06
Low 180 (51.7) 143 (54.6) 37 (43.0)
High 168 (48.3) 119 (45.4) 49 (57.0)
Tumor multifocality, n (%) 0.62
No 178 (51.1) 132 (50.4) 46 (53.5)
Yes 170 (48.9) 130 (49.6) 40 (46.5)
Presence of MP in specimen, n (%) 0.22
No 208 (59.8) 141 (53.8) 67 (77.9)
Yes 140 (40.2) 121 (46.2) 19 (22.1)
Intravesical agents, n (%) 0.50
None 160 (46.0) 124 (47.3) 36 (41.9)
MMC 50 (14.4) 40 (15.3) 10 (11.6)
BCG 113 (32.5) 80 (30.5) 33 (38.4)
Abbreviations: BCG = Bacillus Calmette-Guerin; IQR = interquartile range; MMC = mitomycin C; MP = muscularis propria; RASI =
renin-angiotensin system inhibitor
Maneesuwansin et al.
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TABLE 2. Types of renin-angiotensin system inhibitors.
TABLE 3. Tumor recurrence and stage progression.
Renin-angiotensin system inhibitors n = 86
Angiotensin-converting enzyme inhibitor, n (%) 38 (44.2)
Enalapril 32 (37.2)
Imidapril 1 (1.2)
Nootropril 1 (1.2)
Peridopril 1 (1.2)
Quinapril 3 (3.5)
Angiotensin II receptor blockers, n (%) 48 (55.8)
Irbesartan 5 (5.8)
Losartan 28 (32.6)
Olmesartan 1 (1.2)
Telmisartan 2 (2.3)
Valsartan 12 (14)
All patients No-RASIs RASIs P-value
(n=348) (n = 262) (n = 86)
Tumor recurrence 140 (40.2) 121 (46.2) 19 (22.1) <0.001
Stage progression 19 (5.5) 16 (5.67) 3 (3.5) 0.46
Abbreviations: RASI = renin-angiotensin system inhibitor
Fig 1. Kaplan-Meier curves demonstrates
recurrence-free survival between 2 cohorts of
patients: RASIs (gray) and no-RASIs (black).
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TABLE 4. Univariate and multivariate analyses for factors associated with tumor recurrence.
Univariate analysis Multivariate analysis
HR (95 % CI) P-value HR (95 % CI) P-value
Age ≥ 65 years 1.25 (0.88-1.78) 0.22
Female 0.92 (0.63-1.36) 0.69
Smoking 0.31
Former 0.92 (0.63-1.34)
Current 0.69 (0.43-1.12)
Intravesical 0.26
MMC 1.12 (0.70-1.81)
BCG 0.83 (0.57-1.21)
MMC+BCG 0.55 (0.25-1.20)
High grade 1.16 (0.83-1.61) 0.39
Tumor size ≥ 3 cm 1.56 (1.12-2.19) 0.009 1.50 (1.07-2.10) 0.018
Tumor multifocality 1.53 (1.10-2.14) 0.012 1.49 (1.07-2.09) 0.019
Administration of RASIs 0.45 (0.28-0.73) 0.001 0.46 (0.29-0.75) 0.002
Abbreviations: BCG = Bacillus Calmette-Guerin, CI = condence interval, HR = hazard ratio, IQR = interquartile range, MMC = mitomycin
C, RASI = renin-angiotensin system inhibitor
associated with reduced risks of tumor recurrence and
disease progression in NMIBC patients.
11,12
Of the 348 patients in the present study, 140 (40%)
and 19 (6%) patients experienced tumor recurrence and
progression, respectively. e incidences in our study
were similar to those in research by Yuge et al., which
demonstrated that 39% of patients had tumor recurrence
and 5% had stage progression.
11
On multivariate analysis,
RASI administration was signicantly associated with a
reduced risk of tumor recurrence in our study as well as
in the work of Yuge et al. and Blute et al.
11,12
Nevertheless,
none of the studies showed that RASIs were associated
with disease progression. In our study, the RASI cohort
had a signicantly greater 5-year RFS rate than that of
the no-RASI cohort (78% versus 54%, p=0.001). is
was comparable to the 5-year RFS rates of the study by
Yuge et al., which were 78% and 53% for the RASI and
no-RASI cohorts, respectively, (p=0.01).
11
Patients with NMIBC after a TUR-BT are classified based
on the risk stratication of recurrence and progression.
2-4
Dierent further management strategies are employed
for each group of NMIBC patients. As to the European
Organization for Research and Treatment of Cancer
(EORTC) risk table, patients with multiple or large
tumors are at high risk for bladder cancer recurrence and
stage progression.
2–4,13–15
Our study revealed that tumor
multifocality and a tumor size greater than 3 cm were
associated with an increased risk of tumor recurrence.
Millán-Rodríguez et al. studied a cohort of 1,529 patients
with NMIBC; their Kaplan–Meier analysis demonstrated
that tumor recurrence was statistically signicant for
multiple tumors and a large tumor size (p<0.001). On
multivariate analysis, multifocality and a large tumor
were also associated with an increased risk of tumor
recurrence (OR 2.0, 95% CI 1.6–2.4; and OR 1.7, 95%
CI 1.3–2.0, respectively).
13
It has been suggested that
high-risk NMIBC patients should receive intravesical
BCG to prevent recurrence and progression.
2-4,13-15
Previous meta-analyses have shown that intravesical
therapies signicantly decrease the risk of recurrence
in NMIBC patients.
5
e type of intravesical therapy
used depended on the risk prole of each patient. BCG
therapy showed better outcomes in terms of a reduction
in tumor recurrence than a TUR-BT alone or intravesical
chemotherapy.
5
Nonetheless, some patients could not
tolerate its adverse eects, and some tumors still refracted
or relapsed aer treatment.
6,16
In our study, intravesical
therapy, including BCG and MMC, was not associated
with tumor recurrence and disease progression. e
number of patients recruited for the study could be the
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explanation, given that the role of the RASIs was our
primary outcome.
e major limitation of our study was its retrospective
design with intermediate follow-up. As to the historic
data, the tumor stages and grades diered slightly from the
current classications. While patients given intravesical
BCG in other studies showed a reduced risk of tumor
recurrence and progression, our study had too limited a
number of patients to show similar outcomes. In addition,
supplies of intravesical BCG were unavailable in ailand
during several periods, thereby precluding the utilization of
BCG based upon patient risk classications. Nevertheless,
there was a sucient number of patients in our study
to demonstrate the impact of RASIs on bladder cancer
recurrence. To our knowledge, this is the rst study to
reveal the impact of RASIs on NMIBCs in ailand. A
prospective study will be needed to further evaluate the
eectiveness of RASIs in decreasing NMIBC recurrence
and stage progression.
CONCLUSION
Our study suggests that RASI administration in
patients with NMIBC might be a potential factor to
prevent bladder cancer recurrence. To the best of our
knowledge, this is the rst study in ailand to address
the benets of RASIs for NMIBC patients. Further study
is needed to evaluate the eects of RASIs on NMIBC
patients.
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5. Chou R, Selph S, Buckley DI, Fu R, Grin JC, Grusing S, et
al. Intravesical Therapy for the Treatment of Nonmuscle
Invasive Bladder Cancer: A Systematic Review and Meta-
Analysis. J Urol. 2017;197(5):1189-99.
6. Cambier S, Sylvester RJ, Collette L, Gontero P, Brausi MA, van
Andel G, et al. EORTC Nomograms and Risk Groups for
Predicting Recurrence, Progression, and Disease-specic and
Overall Survival in Non-Muscle-invasive Stage Ta-T1 Urothelial
Bladder Cancer Patients Treated with 1-3 Years of Maintenance
Bacillus Calmette-Guerin. Eur Urol. 2016;69(1):60-69.
7. Sieer-Radtke AO, Apolo AB, Bivalacqua TJ, Spiess PE, Black
PC. Immunotherapy With Checkpoint Blockade in the Treatment
of Urothelial Carcinoma. J Urol.2018;199(5):1129-42.
8. Sparks MA, Crowley SD, Gurley SB, Mirotsou M, Coman TM.
Classical Renin-Angiotensin system in kidney physiology.
Compr Physiol. 2014;4(3):1201-28.
9. Shirotake S, Miyajima A, Kosaka T, Tanaka N, Maeda T, Kikuchi
E, et al. Angiotensin II type 1 receptor expression and microvessel
density in human bladder cancer. Urology. 2011;77(4):1009
e19-25.
10. Tanaka N, Miyajima A, Kosaka T, Miyazaki Y, Shirotake S,
Shirakawa H, et al. Acquired platinum resistance enhances
tumour angiogenesis through angiotensin II type 1 receptor
in bladder cancer. Br J Cancer. 2011;105(9):1331-7.
11. Yuge K, Miyajima A, Tanaka N, Shirotake S, Kosaka T, Kikuchi
E, et al. Prognostic value of renin-angiotensin system blockade in
non-muscle-invasive bladder cancer. Ann Surg Oncol. 2012;
19(12):3987-93.
12. Blute ML, Jr., Rushmer TJ, Shi F, Fuller BJ, Abel EJ, Jarrard
DF, et al. Renin-Angiotensin Inhibitors Decrease Recurrence
aer Transurethral Resection of Bladder Tumor in Patients
with Nonmuscle Invasive Bladder Cancer. J Urol. 2015;194(5):
1214-9.
13. Millan-Rodriguez F, Chechile-Toniolo G, Salvador-Bayarri J,
Palou J, Vicente-Rodriguez J. Multivariate analysis of the
prognostic factors of primary supercial bladder cancer. J
Urol. 2000;163(1):73-78.
14. Rodriguez Faba O, Palou J. Predictive factors for recurrence
progression and cancer specic survival in high-risk bladder
cancer. Curr Opin Urol. 2012;22(5):415-20.
15. Sexton WJ, Wiegand LR, Correa JJ, Politis C, Dickinson SI,
Kang LC. Bladder cancer: a review of non-muscle invasive
disease. Cancer Control. 2010;17(4):256-68.
16. Sfakianos JP, Kim PH, Hakimi AA, Herr HW. e eect of
restaging transurethral resection on recurrence and progression
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341-5.
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Pramote Euasobhon, M.D.*, Suratsawadee Wangnamthip, M.D.*, Chernporn Payomyam, M.D.**, Pranee
Rushatamukayanunt, M.D.*, Sukunya Jirachaipitak, M.D.*, Pratamaporn Chanthong, M.D.***, Janravee
Laurujisawat, RN*, Kesinee Vimolwattanasarn, RN.*
*Siriraj Clinical Pain Management Training Center, Department of Anesthesiology, Faculty of Medicine Siriraj Hospital, Mahidol University, **Division
of Anesthesia, Hua Hin Hospital, Prachubkirikhun 77110, ***Siriraj Palliative Care Center, Faculty of Medicine Siriraj Hospital, Mahidol University,
Bangkok 10700, ailand.
Cancer Pain Management: Is It Still Problematic?
Corresponding author: Suratsawadee Wangnamthip
E-mail: suratsawadee.wan@mahidol.ac.th
Received 12 June 2018 Revised 14 August 2018 Accepted 3 September 2018
ORCID ID: 0000-0002-6795-8550
http://dx.doi.org/10.33192/Smj.2019.07
ABSTRACT
Objective: To evaluate the ecacy of pain management during 3-month follow-ups for outpatients with cancer pain.
Methods: A retrospective chart review was conducted and included all newly diagnosed cancer pain patients
visiting the Siriraj Pain Clinic, Mahidol University, between January 2013 and June 2014. Demographic data, pain
intensity at rst visit, 1-, 2- and 3-month follow-ups, pharmacological therapy and co-treatments were collected.
Good treatment response was dened as more than 30% pain reduction from baseline. Predictive factors associated
with pain treatment response were also assessed.
Results: Out of 432 new patients, 118 cancer pain patients were included in the study with a mean age of 59.8±13.7
years (range 18-91 years). About half of patients had at least one comorbidity. Over 90% of all cancer patients
presented with distance metastasis. Mixed neuropathic/nociceptive pain (53.4%) and nociceptive pain alone (43.2%)
were common pain features in cancer pain patients. e mean initial pain intensity described by verbal numerical
scales was 7.7±2.1 (range 3-10). e majority of patients (60.2%) received co-treatment. e main pharmacological
therapies in all patients were opioids (99.2%) and anticonvulsants (90.7%). At 3-month follow-up, nearly half of
patients achieved a good treatment response. However, 44.6% of good responders still had moderate and severe
pain. No predictive factors associated with the pain treatment response was found.
Conclusion: Approximately half of patients with cancer pain in the pain clinic achieved a good treatment response
whereas one-h of the patients had an increase in pain severity at their 3-month follow-up.
Keywords: Cancer pain; clinical response; pain management; predictive factor (Siriraj Med J 2019;71: 38-43)
INTRODUCTION
Pain is one of the most common symptoms in
cancer patients (70-90%),
1
negatively aecting their
functionality, emotion and quality of life.
2,3
Multiple
factors are involved in the pain mechanism, including the
disease itself, treatments, and comorbidities.
4
Although
cancer pain treatment guidelines were developed by the
World Health Organization (WHO)in the year 1986, and
subsequently modied in 1996,
5
treatment outcomes still
remain unsatisfactory.
6
For example, Deandrea reported
43% of cancer patients suered from moderate to severe
pain.
7
In addition, Yennurajalingam reported that only
45% of cancer patients in the United States receiving
treatment experienced 30% pain improvement on a
numerical rating scale.
8
Regarding the dierence of culture among the nations
Euasobhon et al.
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which may aect to pain acceptance and factor-related to
pain treatment response, the treatment outcome data of
ai patients is required to explore. However, there has
been no study reporting the eectiveness of cancer pain
management services in ailand. erefore, this study
aimed to evaluate the ecacy of cancer pain management
and identify factors associated with the pain treatment
response during 3-month follow-ups at the Siriraj Pain Clinic.
MATERIALS AND METHODS
e study was approved by the Siriraj Institutional
Review Board (Si 703/2014). e researchers retrospectively
and manually reviewed outpatient records of all newly
diagnosed cancer pain patients who visited Siriraj Pain
Clinic from January 1
st
, 2013 to June 30
th
, 2014. Cancer
patients with non-cancer pain conditions and patients who
were lost before the 3-month follow-up were excluded.
Demographic data including age, gender, living region,
comorbidities (diabetes mellitus, hypertension, heart
disease and chronic kidney disease), primary cancer site
(gastrointestinal tract, respiratory tract, urogenital tract,
breast, etc.), pharmacological therapy and co-treatments
were recorded. Pain intensity using verbal numerical
scales (VNS) at rst visit, 1-, 2- and 3-month follow-ups
were collected as the primary outcomes.
Pain severity was classied into three levels which
were mild pain (VNS 0-3), moderate pain (VNS 4-6)
and severe pain (VNS 7-10).
Responses to treatments varied from time to time,
even for individual patients. e timing of pain evaluation
was crucial. If pain was evaluated aer a few weeks
following treatment, the number subjects in the non-
responding group may be appear elevated because it
could be too early to see a response. Conversely, it is
possible evaluating pain at distant time periods could
allow the subjects’ cancer to progress and cause more
pain, reducing the number in subjects the responsive
group. erefore, the optimum time for evaluating the
ecacy of pain treatment in this study was set at three
months from the initial treatment. Factors associated
with pain treatment responses were also analyzed.
Statistical analysis
e sample size was calculated based on the study of
Yennurajalingam et al
8
and required 40% proportion of
patients to achieve good treatment response at 3-month
follow-up. We selected the condence level 95% and
allowable error 10% to calculate by Query program.
Calculated sample size was 93 patients with 10% dropout.
erefore, at least 102 patients were included in this
study.
e qualitative data, including gender and main site of
cancer, was described as numbers and percentages. e
quantitative data, consisting of the initial pain scores,
numbers of visits,and age, was described using mean
and standard deviation.
According to Farrar’s study, the researchers dened
good treatment response as the pain reduction of at least
30% using a verbal numerical scale.
9
To compare the
good treatment response group and the poor treatment
response group (VNS pain responses less than 30%),
unpaired t-test (normality) or Mann-Whitney U-test
(non-normality) were used to analyze quantitative data
and Chi-square test or Fisher’s exact test were used for
qualitative data. Logistic regression analysis was applied
when there were one or more statistically signicant
factors (p < 0.05). All parameters were analyzed by SPSS
program version 15.0.
RESULTS
Aer screening outpatient charts, 432 cancer pain
patients were reviewed; of these 314 were excluded
due to attrition or incomplete follow-ups during the 3
months. A total of 118 cancer patients were included
in this study (Table 1), 50.8% of which were males with
the median age of 60 years old (range 18-91 years old).
About half of the patients had at least one comorbidity.
Most of the patients presented with advanced stage of
cancer, accounting for 91.5% of the total number. e
majority of patients suered from nociceptive and mixed
pain. e most common adjuvant treatment (apart from
pain medication) was radiation (28.8%) while only two
patients underwent the pain intervention. Regarding
pharmacological treatment, almost all of the patients
(117 patients) were prescribed opioids (Table 2). e
most common opioid used was morphine (54.7%), while
the most common adjuvant drugs were anticonvulsants
(90.7%). Dosages for opioid therapy in each visit have
been presented in Table 3.
e overall outcome aer cancer pain treatment
found that the mean VNS decreased from 7.7±2.1 to
5.7±2.6 at the 1-month follow-up and maintained at this
level until 3-month (Table 3). Moreover, there were 47.5%
of the patients achieving good treatment response while
44.6% of which still suered from moderate to severe
pain at three months aer treatment (Fig 1). However,
19.5% of the patients got worse even aer 3 months of
pain treatment.
Many potential factors associated with treatment
outcome were analyzed, including age, gender, comorbidity,
primary organ tumour, stage of cancer, initial pain
intensity, types of pain, adjuvant therapy, type and dose
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TABLE 1. Demographic data.
TABLE 2. Medications used in pain treatment on 3-month follow-up visit.
Patient characteristics N=118
Age (yrs) 60 (18-91)
Gender M:F 60 (50.8):58 (49.2)
Comorbidity
0 62 (52.5)
1 34 (28.8)
≥ 2 22 (18.6)
Primary tumor
Gastrointestinal system 26 (22.0)
Respiratory system 20 (16.9)
Others 72 (61.0)
Stage of cancer
Local invasion 10 (8.5)
Distant metastasis 108 (91.5)
Initial pain intensity (verbal numerical scale) 8 (3-10)
Types of pain
Nociceptive 51 (43.2)
Neuropathic 4 (3.4)
Mixed pain 63 (53.4)
Co-treatment
Radiation therapy 34 (28.8)
Chemotherapy 24 (20.3)
Surgery 10 (8.5)
Pain intervention 2 (1.6)
Data are presented as median (range), n (%)
Drugs N=118
Opioids 117 (99.2)
Strong opioid 78 (66.1)
morphine 64 (54.7)
fentanyl 12 (10.3)
methadone 2 (1.7)
Weak opioid 39 (33.1)
codeine 4 (3.4)
tramadol 35 (29.9)
Acetaminophen 49 (41.5)
NSAIDs 37 (31.4)
Antidepressant 70 (59.3)
Anticonvulsant 107 (90.7)
Data are presented as n (%), NSAIDs = Non-steroidal anti-inammatory drugs
Euasobhon et al.
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TABLE 3. Opioid dosage and average pain intensity in each visit for 3 months.
Opioid 1
st
visit 1-month 2-month 3-month
Morphine (mg/day) 20 (10-180) 20 (10-200) 20 (10-180) 35 (10-180)
Fentanyl (mcg/hr) 12 (12-25) 25 (12-100) 25 (12-50) 25 (12-37)
Methadone (mg/day) 10 (10-10) 10 (10-15) 20 (10-30) 10 (10-10)
Codeine (mg/day) 60 (45-90) 60 (45-90) 45 (45-60) 45 (30-60)
Tramadol (mg/day) 150 (50-400) 150 (50-300) 150 (100-300) 150 (100-200)
VNS 7.7±2.1 5.7±2.6 5.8±2.9 5.7±3.1
Data are presented as median (range), mean±SD
Abbreviation: VNS = verbal numerical scales
of opioids, etc. However, there were no predictive factors
signicantly associated with the pain treatment response
(Table 4).
DISCUSSION
This study evaluated outcomes of cancer pain
treatment at the Siriraj Pain Clinic at three months
aer treatment. Even though aer following regimens
provided by guidelines for cancer pain management
5
,
average pain score was reduced from 7.7 to 5.7 which
was about 26% pain reduction. Also, 42.3% of patients
were still in severe pain and less than half achieved good
treatment response. However, this data was comparable
to Yennurajalingam’s study, in which the responsive
group was accounted for 45% of subjects.
8
Regarding
the treatment outcome in this study revealed that cancer
pain is still problematic even in the tertiary pain center.
Considering about one-fifth of patients in this
study and one-third of patients in the Yennurajalingam’s
study developed worse pain over 3 months,
8
this could
be attributed to either rapid cancer progression or the
duration of the visits per month was perhaps too long,
particularly in advanced stage cancer patients. erefore,
more frequent follow-up visits should be set in the case
of uncontrolled pain. To address some of the diculties
that were encountered, for example, remoteness from the
hospital or physical limitations, a telephone interviewing
program is advisable. In addition, some patients might
not well tolerated to higher dose of opioids or doubted
about using opioid therapy which could be another
hindrance to increase opioid dosage. Interestingly, mean
opioid dosage appeared to be stable from the rst visit
to 3-month visit, which should have been increased in
the following visits if the pain could not be controlled
Fig 1. Pain treatment response at 3-month follow-up.
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TABLE 4. Factors associated with pain treatment outcomes.
Factors
Good treatment response Poor treatment response
P-value
n=56 n=62
Age 58.7±13.5 60.8±14.0 0.418
Female 28 (50) 30 (48.4) 0.861
Living outside Bangkok and metropolitan area 27 (48.2) 35 (56.5) 0.371
Comorbidities 25 (44.6) 31 (50) 0.561
Primary organ tumor
Gastrointestinal system 12 (21.4) 14 (22.6)
Respiratory system 9 (16.1) 11 (17.7) 0.949
Others 35 (62.5) 37 (59.7)
Advanced cancer stage 50 (89.3) 58 (93.5) 0.515
Initial pain intensity 8.0±2.1 7.4±2.1 0.976
Types of pain
Nociceptive 28 (50) 23 (37.1)
Neuropathic 2 (3.6) 2 (3.2) 0.348
Mixed 26 (46.4) 37 (59.7)
Co-treatment
Radiation therapy 13 (23.2) 21 (33.9) 0.202
Chemotherapy 13 (23.2) 11 (17.7) 0.461
Surgery 4 (7.1) 6 (9.7) 0.746
Pain intervention 0 (0) 2 (3.2) 0.497
Opioid dosage
Morphine 45.0±36.5 46.8±35.3 0.803
Fentanyl 24.8±8.8 28.3±9.4 0.492
Tramadol 145±35.9 153±35.2 0.793
Data are presented as mean±SD, n (%)
(Table 3). Nevertheless, the poor treatment response
group seemed to consume only a little more opioids
than the good response group (Table 4), presenting
that opioids might not help in the poor response group
or they experienced some problems to increase opioid
consumption.
e previous study found that high initial pain intensity
was a factor associated with good clinical response in
outpatients.
8
In contrast, another retrospective study in
inpatients demonstrated that high pain intensity on the
rst day of consultation was related to poor treatment
response.
10
However, there was no association between
initial pain intensity and the pain treatment response in
this study. In addition, no signicant association of any
potential factors to the treatment response was observed.
Although this study was a retrospective study, all
data were obtained by pain specialists and well-trained
nurses in pain management, which made the data more
reliable. However, there were some limitations in this
study. Firstly, some data were not documented in some
included charts, for instance, the side eects, the frequency
of incidental pain or eects of pain on emotion and
function, which may aect the treatment outcomes.
erefore, we decide not to present and analyze those data
in this study. Secondly, the number of patients may not
be enough to detect signicant factors associated with the
Euasobhon et al.
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treatment response. Although there were a large number
of cancer patients consulted the pain clinic, about 73%
of reviewed charts were excluded from this study mainly
due to incomplete follow-up in three months, which
may cause by patients’ death within three months aer
the rst visit or some diculty of traveling to-and-from
the hospital. Despite the limitations, the results of this
study serve as baseline data for cancer pain treatment
and further development of guidelines for cancer pain
management.
Future prospective studies should be conducted to
compare the eectiveness of treatments between dierent
cancer service centers. However, future prospective
research with cancer pain management protocol should
be developed to improve the eectiveness of cancer
pain treatment. Also, the psychological aspects played
an important role in cancer patients and could not
be solved only by pain medications. Multidisciplinary
teams, including psychiatrists and behavioral therapists,
could oer better care and help improve the treatment’s
ecacy.
CONCLUSION
Approximately half of patients with cancer pain
in the pain clinic achieved a good treatment response
whereas one-h of the patients had an increase in pain
severity at 3-month follow-up. ere is no signicant
factor that associated with treatment response.
ACKNOWLEDGMENTS
It is very grateful to Somthawil Potranan and
Phantip Phongwanichanan, pain specialist nurses, for
the excellent help in data gathering. e authors would
also like to thank Nichapat Sooksri and Sunsanee Mali-
ong, research assistants, for administrative work and
Suthipol Udompunthurak for his statistical analysis
assistance.
Funding: Siriraj Research Development Fund, Faculty
of Medicine Siriraj Hospital, Mahidol University.
Potential conicts of interest: None.
REFERENCES
1. Portenoy RK, Lesage P. Management of cancer pain. Lancet.
1999;353:1695-700.
2. Cleeland CS, Gonin R, Hateld AK. Pain and its treatment in
outpatients with metastatic cancer. N Engl J Med. 1994;330:592-
96.
3. Breivik H, Cherny N, Collett B. Cancer-related pain: A pan-
European survey of prevalence, treatment, and patient attitudes.
Ann Oncol. 2009;20:1420-33.
4. Mellar PD, Declan W. Epidemiology of cancer pain and factors
inuencing poor pain control. Am J Palliat Med. 2004;21:137-
42.
5. Zech DF, Grond S, Lynch J, Hertel D, Lehmann KA. Validation
of World Health Organization Guidelines for cancer pain
relief: a 10-year prospective study. Pain. 1995;63:65-76.
6. Manfredi PL, Chandler S, Pigazzi A, Payne R. Outcome of
cancer pain consultations. Cancer. 2000;89:920-24.
7. Deandrea S, Montanari M, Moja L, Apolone G. Prevalence
of under treatment in cancer pain. A review of published
literature. Ann Oncol. 2008;19:1985-91.
8. Yennurajalingam S, Kang JH, Hui D, Kang DK, Kim SH, Bruera
E. Clinical response to an outpatient palliative care consultation
in patients with advanced cancer and cancer pain. J Pain Sym
Man. 2012;44:340-50.
9. Farrar JT, Young JP Jr, LaMoreaux L, Werth JL, Poole RM.
Clinical importance of changes in chronic pain intensity
measured on an 11-point numerical pain rating scale. Pain.
2001;94:149-58.
10. Wangnamthip S, Euasobhon P, Siriussawakul A, Jirachaipitak
S, Laurujisawat J, Vimolwattanasarn K. Eective Pain Management
for Inpatients at Siriraj Hospital: A Retrospective Study. J Med
Assoc ai. 2016;99:565-71.
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44
Sapol epwiwatjit, M.D., Sasiriyar Athisereerusth, M.D., Wanicha Lertpipopmetha, anit Nanthanasub,
M.D., Yodying Dangprapai, M.D., Ph.D.
Department of Physiology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok 10700, ailand.
Patient Interviews Improve Empathy Levels of
Preclinical Medical Students
Corresponding author: Yodying Dangprapai
Email: yodying.dan@mahidol.ac.th
Received 3 April 2018 Revised 19 October 2018 Accepted 7 November 2018
ORCID ID: 0000-0003-4437-2351
http://dx.doi.org/10.33192/Smj.2019.08
ABSTRACT
Objective: In order to cultivate and maintain empathy during medical school, an experiential learning program, “A
Patient as a Human Being”, was designed to promote empathy in second-year medical students through interviews
with patients focusing on their suering and the diculties arising from their illnesses and hospital stays.
Methods: e second-year medical students were divided into groups of three and four. Each group was assigned
a patient to interview under close supervision. e selected patients were informed beforehand about the interview
and voluntarily agreed to participate. e ai version of the Jeerson Scale of Physician Empathy–Student Version
(JSPE-SV) was used to assess the students’ empathy levels.
Results: e baseline JSPE-SV score (n = 310) was 114.10±10.20. Aer the interview, the scores signicantly increased
(1.19 [0.21-2.18], P = 0.009). Students in the lower-half group of baseline scores showed a higher improvement (2.64
[1.14–4.15], P < 0.001) than those in the upper-half group. e dierence coecient by multivariate analysis of the
improved JSPE–SV scores between the two groups was 3.03 [1.08-4.98] (p = 0.002), accompanied by a correlation
between the pre-activity empathy score and the improved score ( p = - 0.21, P-value < 0.01).
Conclusion: e patient interviews improved the empathy levels of the preclinical medical students, especially
those with lower baseline empathy levels.
Keywords: Empathy; preclinical medical student; experiential learning; patient interview (Siriraj Med J 2019;71: 44-51)
INTRODUCTION
Empathy has been described as the ability to understand
and accept the feelings of other people.
1-3
In medicine,
empathy may be dened as the capability of physicians
to accurately comprehend the mental status of their
patients.
1
Importantly, empathy has been described as
one of the important components of the doctor-patient
relationship in the provision of holistic patient care.
4–8
Nevertheless, the empathy level of medical students tends
to decline during medical school.
8-14
erefore, empathy
maintenance and improvement during a course of medical
training has been emphasized.
5,7, 15-18
Various activities
have been created to enhance empathy among medical
students, including communication-skill training in various
formats, such as didactic lectures, experiential learning,
role-playing, and reective writing aer a learning session
with standardized patients. However, students’ empathy
levels were not measured as part of those activities.
19, 20
To study empathy in medicine, various questionnaires
have been developed as a measurement tool, such as
the Jeerson Scale of Physician Empathy (JSPE)
21
, the
Empathy Test (ET)
22
, the Balanced Emotional Empathy
Thepwiwatjit et al.
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Scale (BEES)
23
, and the Consultation and Relational
Empathy (CARE).
24
Among those tools, the JSPE is the
most widely used instrument for assessing the empathy
of healthcare professionals, including medical students.
e JSPE–Student Version (SV) has been developed and
validated into many languages, including ai.
To cultivate empathy in preclinical medical student,
the development program entitled “A Patient as a Human
Being” was designed as a mandatory activity in the
Humanistic Medicine course for second-year medical
students. rough a group interview of a patient admitted
to Siriraj Hospital, students focused on the patient’s
suering and the diculties arising from the illness and
hospital stay. Aer the interview, each student submitted
a piece of reective writing exploring the interview
experience. To evaluate the eects of this experiential
learning on students’ empathy, the ai version of the
JSPE–SV was utilized.
MATERIALS AND METHODS
Participants
e ai medical school system has implemented
a 6-year curriculum for the bachelor’s degree in Doctor
of Medicine (M.D.). e rst year, referred to as the pre-
medicine year, focuses on general education and/or the
liberal arts. e next two years are the preclinical years,
and the remainder of the course is the clinical years. e
present study focused on preclinical medical students at
the Faculty of Medicine Siriraj Hospital, in particular,
the second-year medical students (n = 325) participating
in the activity “A Patient as a Human Being”, which is a
part of the compulsory Humanistic Medicine course. In
all, 310 students (n = 310) voluntarily participated in the
study. However, 14 of those students were subsequently
excluded because their questionnaires were incomplete.
Instrument
e ai version of the Jeerson Scale of Physician
Empathy–Student Version (JSPE–SV)
25
was used to evaluate
the empathy level of each medical student before and
aer participating in the activity “A Patient as a Human
Being”. e JSPE–SV comprises twenty self-reported
statements. Students rated each statement from one to
seven, which represented the spectrum from “strongly
disagree” to “strongly agree”, respectively. e total score
could range from 20 to 140 points. Participants with a
higher score were regarded as having a higher level of
empathy than those with a lower score. e development
of the ai version of the JSPE–SV by the back-translation
procedure had been previously examined, and it had
demonstrated acceptable validity and reliability.
25
Intervention
“A Patient as a Human Being” is one of the learning
activities in the Humanistic Medicine course for second-
year medical students at the Faculty of Medicine, Siriraj
Hospital. It provides them with rst-hand experience in
interviewing a patient regarding the suering arising
from having a disease and the diculties associated with
a hospital stay. In other words, this activity provides
the opportunity for the preclinical medical students to
explore and to feel suering and hardship from a patient’s
perspective. Before the activity, the professional manners
to be employed during the interview were introduced.
Each group of three or four medical students subsequently
met with an assigned faculty member and a h-year
medical student to prepare for the patient interview.
e recruitment of each selected inpatient was based
on voluntary permission to participate in the activity.
A thirty-minute interview was conducted by a group of
preclinical students under the supervision of a senior
medical student or faculty member. Aer the interview,
the second-year students reected on their experiences
through conversation with an assigned faculty member
and reective writing.
To investigate the eects of the activity on the
empathy levels of second-year medical students, a study
protocol was submitted to, and approved by the Siriraj
Institutional Review Board (Si 759/2016). Details of the
research were subsequently explained to second-year
medical students. Each student who voluntarily decided
to participate later anonymously completed the ai
version of the JSPE-SV. is was done before and aer
their patient interview, using a web-based questionnaire
created by the Undergraduate Medical Education Unit
of the Faculty of Medicine, Siriraj Hospital. e empathy
scores before and aer the interview and reection tasks
were designated as the pre-activity and post-activity
scores, respectively.
Data analysis
Data acquired from the electronic database consisted
of the pre- and post-activity scores, and demographic
information such as gender and grade point average
(GPA). ese underwent a quantitative analysis using
the R Statistical System, version 3.2.5, with the built-in
library and signicance threshold set to 0.05.
To determine the eects of the interview-reection
activity on the empathy scores, the pair-wised dierence
between the pre-activity and post-activity scores (which
represents improvement) was examined with the one-
tailed dependent t-test. To determine the factors associated
with an improvement in the empathy scores, further
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46
analyses were performed on subgroups of students in
terms of their gender, GPA, participation in university
activities, experience in taking care of patients with chronic
illnesses, and their having heard the word “empathy”
before admission to the medical school.
To transform the GPA and the pre-activity score from
numerical data to categorical data, the median cut-point
was applied to classify these samples into two subgroups.
For instance, with the “Higher GPA” vs. the “Lower GPA”
subgroups, the cut point was the median of the GPAs of
the participants. As a further example, in the case of the
“Higher pre-activity score” vs. the “Lower pre-activity
score” subgroups, the cut point was the median of the
pre-activity scores of the participants. Since the data
were categorized into at most two subgroups for each
variable, the dierence between the subgroups for each
variable was evaluated by the two-tailed independent
t-test. e Pearson correlation coecient was applied
to evaluate any association between the numerical factor
and the pre-activity empathy score.
To study the factors determining the pre-activity
empathy scores, a comparison was made of the score for
each subgroup within the same variable (e.g., the “Higher
GPA” vs. the “Lower GPA” subgroups). A univariate
analysis was performed on each variable using a two-tailed
independent t-test for categorical factors. Multivariate
analyses were performed on the between-subgroup
dierence using a multivariate linear regression model
to adjust for confounding eects.
RESULTS
Of the 325 second-year medical students, 310 students
(95.38%) undertook the web-based questionnaire, with
296 (91.08%) of the questionnaires being fully completed.
e 14 students with incomplete data were excluded
from this study. e demography of the categorical
and numerical variables is presented in Tables 1 and 2,
respectively.
Association factors determining pre-activity scores
Unadjusted associations between the pre-activity
empathy scores and the candidate variables demonstrated
no statistically signicant dierences. e detailed results
of the association factors that determined the pre-activity
scores are in Table 3.
Improvements in empathy scores
Overall, the statistical analyses revealed a signicant
increase in the empathy scores from the pre- to the post-
activity scores (1.19 [0.21–2.18], P-value = 0.009). e
unadjusted comparisons within the subgroups showed
signicant improvements in the empathy scores of the
following subgroups: male students (1.31 [0.01–2.62],
P-value = 0.025); students with a higher GPA (1.60 [0.35–
2.85], P-value = 0.007); students with no participation in
the extracurricular activities of the Mahidol University
(2.06 [0.25–3.87], P-value = 0.014); students who had
experience of taking care of a patient with chronic
illness (1.84 [0.37–1.31], P-value = 0.008); students
with no experience of hearing the word “empathy”
before admission to the medical school (1.81 [0.11–3.51],
P-value = 0.020); and students with a lower pre-activity
empathy score (2.64 [1.14–4.15], P-value < 0.001). When
the improved empathy scores were compared between
the subgroups, the only statistically signicant factor was
the pre-activity score. From the multivariate analysis,
the students in the “Lower pre-activity” group had a
significantly higher improvement in empathy score
than those in the “Higher pre-activity” group (P-value
= 0.002). When the GPA and pre-activity empathy score
were considered as a scale variable, there were signicant
degrees of correlation between the improvement and GPA
(p = 0.13, P-value = 0.03) and between the improvement
and pre-activity empathy score (p = - 0.21, P-value
< 0.01). ese correlations supported the ndings of
the categorical groups’ comparison. e details of the
statistical analyses are given in Tables 4 and 5.
DISCUSSION
“A Patient as a Human Being”, a truly experiential
learning activity that improves the empathy levels of
preclinical medical students, involves authentic patients.
is distinguishes it from previously reported activities
using simulated medical consultations with standardized
patients
26
or clinical interview training with simulated
situations
27
rough direct communication with authentic
patients, “A Patient as a Human Being” oers a unique
learning experience designed to cultivate empathy among
preclinical students participating in an otherwise traditional
medical curriculum. Well-developed communication
skills are essential for a doctor to express empathy and
to give high quality care.
28-30
Conversations with real
patients have previously been reported to increase and
maintain the empathy levels of medical students.
31,32
e JSPE is an instrument that has been used by
many previous studies to measure levels of empathy
as well as to evaluate the eects of learning activities
designed to improve and maintain empathy
13,15,26,33-35
Importantly, the JSPE has been translated into ai and
validated on ai medical students.
25
erefore, the ai
version of the JSPE was the best available tool to study
empathy levels in ailand.
Thepwiwatjit et al.
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TABLE 1. Demographic information of the numerical data.
Numerical variables Min. - Max. Mean S.D.
Age (years) 18 - 21 19.58 0.59
Grade point average 2.13 - 4.00 3.48 0.33
Pre-activity empathy score 76 - 140 114.10 10.20
Post-activity empathy score 72 - 140 115.30 11.88
Abbreviations: min. = the minimum value, max. = the maximum value, S.D. = standard deviation of the mean
TABLE 2. Demographic information of the categorical data.
Categorical variables N (% of total)
Gender
Male 181 (61.15)
Female 115 (38.85)
Grade point average
High (Mean = 3.75, S.D. = 0.13) 149 (50.34)
Low (Mean = 3.20, S.D. = 0.24) 147 (49.66)
Participation of university activities
Yes 183 (61.82)
No 113 (38.18)
Experience in caring for a patient with chronic illnesses
Yes 135 (45.61)
No 161 (54.39)
Having heard the word ‘empathy’ before admission to the medical school
Yes 186 (62.84)
No 110 (37.16)
Abbreviations: N = numbers of subject, S.D. = standard deviation of the mean
In this study, the baseline empathy score of the
second-year medical students was 114.10±10.20, which
was similar to the mean score of the second year medical
students at another medical school in ailand.
25
e
Faculty of Medicine, Siriraj Hospital, introduces the word
“empathy” to rst-year medical students through the
mandatory course entitled Medical Profession. erefore,
the term “empathy” was not considered a new word for
the second-year medical students.
Previous studies have demonstrated associations
between empathy levels and factors such as gender and
education level. For example, female students have been
shown to have a higher mean empathy score than male
students.
14,15, 25, 35, 36
In contrast, in the current study, the
relatively limited number of female medical students
might have been responsible for there being no signicant
dierence in the empathy levels of the genders. In addition,
it has been demonstrated that higher education levels
result in higher empathy scores.
37
However, a high GPA
has also been associated with lower empathy scores.
38
Furthermore, it has been demonstrated that medical
students attending any service activity (such as the free
clinic for patients without insurance) have a higher
empathy score than those who have never attended.
35
In contrast to the ndings of the other studies,
the present study did not nd any association between
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48
TABLE 3. Associations of the pre-activity empathy score with corresponding subgroup analysis for each candidate
variable.
Variables N Mean S.D. Between
Lower 95% CI Upper 95% CI
P-value
Subgroup
Difference
Gender 0.29 -2.07 2.65 0.808
Male 181 114.20 10.55
Female 115 113.91 9.73
Grade point average 0.21 -2.13 2.55 0.861
High 149 114.19 10.89
Low 147 113.99 9.53
Participation of university activities - 0.17 - 2.54 2.20 0.889
Yes 183 114.03 10.53
No 113 114.19 9.75
Experience in taking care of a patient - 0.79 - 3.12 1.53 0.501
with chronic illnesses
Yes 135 113.66 9.47
No 161 114.45 10.83
Having heard the word ‘empathy’ 0.71 - 1.73 3.15 0.567
before admission to the medical school
Yes 186 114.35 10.14
No 110 113.65 10.39
Abbreviations: N = numbers of subject, CI = Condence Interval
the empathy scores and gender, GPA, participation
in university activities, experience of taking care of a
patient (e.g., a family member) with chronic illnesses,
or having heard the word “empathy” before admission
to the medical school. According to the current study,
for each subgroup comparison (Table 3), male gender,
having a higher GPA, the non-participation in university
activities, no experience of taking care of a patient with
chronic illnesses, and having heard the word “empathy”
before medical school admission showed a positive trend
to having a higher baseline empathy score. However,
the dierences in the empathy scores of all factors were
without statistical signicance.
“A Patient as a Human Being” was designed to
improve and maintain the empathy levels of preclinical
medical students. From the empathy scores measured
by the JSPE–SV, ai version, the student participants
in the present study demonstrated an improvement of
1.19 [0.21–2.18] in the eect size, and with statistical
signicance (P = 0.009). Many activities
19
have been
reported by other studies to improve empathy levels,
for example, simulated medical consultations using
standardized patients with subsequent reection by
students on the patients’ feelings about their diseases and
the doctors’ feelings towards the patients.
26
In addition,
participation in communication skills workshops
27
has
also been demonstrated to improve empathy levels.
Even though there was not much improvement in the
scores, the current study strengthened the nding of
many previous studies that experiential learning gained
from authentic situations, simulations, or workshops is
eective in improving empathy.
“A Patient as a Human Being” had a statistically
signicantly greater impact on the students who had a
lower baseline empathy level than those with a higher
level. e students with the lower baseline empathy level
had a statistically signicant improvement in eect size
of 2.90 by univariate analysis and 3.03 by multivariate
analysis. is study demonstrated that a medical school
could improve the empathy levels of preclinical medical
students through a patient interview and subsequent
reection on the patient’s suering. Moreover, a previous
study found that medical students with a higher empathy
level demonstrated a lower rate of empathy decrease than
those with a lower baseline empathy level
14
erefore,
medical schools could sustain the empathy levels among
students by providing an interview-reection activity
throughout their curriculum.
Thepwiwatjit et al.
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TABLE 4. Univariate analysis of within subgroup improvement of the empathy score aer participation in ‘A patient
as human being’ activity. Statistical analyses of empathy score within subgroup improvements and corresponding
univariate subgroups were demonstrated.
Categorization N Within Lower 95% CI Upper 95% CI P-value
subgroup
improvement
Overall 296 1.19 0.21 2.18 0.009
By Gender
Male 181 1.31 0.01 2.62 0.025
Female 115 1.01 - 0.46 2.48 0.091
By GPA
Higher 149 1.60 0.35 2.85 0.007
Lower 147 0.78 - 0.73 2.30 0.157
By experience of participation in university
activities
Yes 183 0.66 - 0.47 1.80 0.127
No 113 2.06 0.25 3.87 0.014
By experience of taking care of a patient
with a chronic illness
Yes 135 1.84 0.37 3.31 0.008
No 161 0.65 - 0.66 1.97 0.166
By experience of hearing the word ‘empathy’
before admission
Yes 186 0.83 - 0.36 2.02 0.088
No 110 1.81 0.11 3.51 0.020
By pre-activity empathy score
High score 148 - 0.26 - 1.47 0.96 0.660
Low score 148 2.64 1.14 4.15 < 0.001
Abbreviations: N = numbers of subject, CI = Condence Interval
In the subgroup division of an unadjusted comparison
(Table 5), differences between each subgroup were
demonstrated using both univariate and multivariate
analyses. In the case of the male medical students, a lower
baseline empathy level, a high GPA, and no participation
in university activities showed an improvement in empathy
scores aer participating in “A Patient as a Human
Being”. ese observations imply that the benets of this
activity were not homogenous among medical students
from dierent subgroups. In agreement with previous
studies, there would appear to be no single activity
capable of improving the empathy levels of students from
diverse backgrounds, but the personal and professional
development activities (such as communication skills
workshops, interpersonal skills workshops, and literature
and medicine programs) would help students realize the
importance of empathy in the medical profession.
36
One of limiting factors of this study was the dierences
among the patients participating in the interviews. To
illustrate, some patients shared their stories with humor,
whereas others were highly emotional, expressing their
feelings tearfully through a large part of the interview.
Furthermore, through the self-reective writing, it was
apparent that students from the same interview session
demonstrated dierent degrees of perception regarding
patients’ suerings and diculties. Another possible
limitation could be that “A Patient as a Human Being” is
an intra-curricular activity. erefore, there was no control
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50
group to emphasize the true eect of this experiential
learning. Last but not least, this study demonstrated
only the short-term eect of the patient interviews and
subsequent reection; no long-term assessment was
made.
CONCLUSION
“A Patient as a Human Being”, a mandatory learning
activity in a traditional six-year medical curriculum,
provided rst-hand experience for the preclinical medical
students to feel patients’ suering from their current
illnesses and hospital stays. Aer the interviews with
the patients, through an experiential learning cycle,
students reected on what they had learned during
the interviews and how they could help the patients as
preclinical medical students. Participation in “A Patient
as a Human Being” increased the second-year medical
students’ empathy levels, especially in the case of students
with lower baseline empathy levels.
TABLE 5. Univariate and multivariate analysis of between subgroup dierence of the empathy score aer participation
in ‘A patient as human being’ activity. Statistical analyses of empathy score between subgroup and corresponding
univariate and multivariate subgroups were demonstrated.
Categorization Univariate Analysis Multivariate Analysis
Difference* Lower Upper P-value Difference* Lower Upper P-value
95%CI 95%CI coefcient 95%CI 95%CI
By Gender
Male > Female 0.30 - 1.68 2.28 0.765 0.41 - 1.60 2.42 0.691
By GPA
Higher > Lower 0.82 - 1.16 2.79 0.417 1.24 - 0.73 3.20 0.217
By experience of participation in
university activities
No>Yes 1.41 - 0.74 3.55 0.197 1.44 - 0.59 3.47 0.164
By experience of taking care of
a patient with a chronic illness
Yes>No 1.19 - 0.80 3.16 0.240 1.64 - 0.34 3.61 0.104
By experience of hearing the word
‘empathy’ before admission
No>Yes 0.98 - 1.12 3.07 0.356 0.97 - 1.06 2.99 0.348
By pre-activity empathy score
Low score > High score 2.90 0.95 4.84 0.004 3.03 1.08 4.98 0.002
Note: Dierence* refers to between subgroup dierence
Abbreviation: CI = Condence Interval
ACKNOWLEDGMENTS
e authors would like to thank the Department of
Psychiatry, Faculty of Medicine Ramathibodi Hospital,
Mahidol University for the ocial permission to use the
ai version of the Jeerson Scale of Physician Empathy-
Student Version (JSPE-SV). Furthermore, we would like
to thank Associate Professor Rungnirand Praditsuwan,
the Vice Dean for Undergraduate Education, the Faculty
of Medicine Siriraj Hospital for his invaluable advices
throughout this study.
Funding: is study was supported by an Education
Research Fund from the Faculty of Medicine Siriraj
Hospital, Mahidol University.
Declaration of interest: e authors have no conict
of interest.
Thepwiwatjit et al.
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Varut Lohsiriwat, M.D., Ph.D., Panumat Jaturanon, M.D.
Department of Surgery, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok 10700, ailand.
Effect of Intraoperative Hypothermia on Surgical
Outcomes after Colorectal Surgery within
an Enhanced Recovery after Surgery Pathway
Corresponding author: Varut Lohsiriwat
E-mail: bolloon@hotmail.com
Received 6 July 2018 Revised 22 July 2018 Accepted 6 November 2018
ORCID ID: 0000-0002-2252-9509
http://dx.doi.org/10.33192/Smj.2019.09
ABSTRACT
Objective: e adverse eects of intraoperative hypothermia from the published literature were mainly based on non-
enhanced recovery aer surgery (ERAS) settings. is study aimed to determine association between intraoperative
hypothermia and outcomes following colorectal surgery under ERAS pathway.
Methods: A prospectively collected database of patients undergoing elective colorectal surgery under ERAS pathway
from 2011 to 2015 was reviewed. Patients were divided into 2 groups: hypothermic group (core temperature <36
o
C
continuously exceeding 30 minutes during an operation) and normothermic group. Short-term outcomes were
compared.
Results: is study included 195 patients: 150 (77%) in hypothermic group and 45 (23%) in normothermic group.
Rectal surgery (OR=5.15), operative time exceeding 3 hours (OR=3.80), multi-organ resection (OR=3.12) and male
gender (OR=2.62) were signicant predictors for intraoperative hypothermia. Rates of postoperative complication
and wound infection were comparable between hypothermic patients and normothermic patients (23% vs 13%;
p=0.17 and 6.0 vs 6.7%; p=0.87, respectively). Hypothermic patients had a longer time to tolerate normal diet (2.0
days vs 1.3 days; p=0.023) but a comparable time to rst bowel movement (2.6 days vs 2.6 days; p=0.84). Hypothermic
patients had a signicant longer hospitalization (5.7 days vs 4.4 days; p=0.048). A multivariate analysis showed
that intraoperative hypothermia was an independent predictor for delayed food intake (OR=2.9, 95%CI=1.2-6.9;
p=0.014) but not for prolonged hospitalization (OR=1.7, 95%CI=0.7-3.9; p=0.207).
Conclusion: Intraoperative hypothermia prolonged time to tolerate food intake aer colorectal surgery within an
ERAS setting but it did not adversely aect the return of bowel function, wound infection, complication and length
of hospitalization.
Keywords: Hypothermia; enhanced recovery aer surgery; colon; rectum; surgery; outcomes (Siriraj Med J 2019;71:
52-58)
INTRODUCTION
During an intraabdominal operation, patient’s
body temperature decreases as a result of impaired
thermoregulatory mechanisms secondary to anesthesia and
e abstract of this manuscript was presented as a poster at the Annual Scientic Congress of the Royal Australasian
College of Surgeons, Australia, between 8 May and 12 May 2017.
heat loss through a surgical wound or to the environment.
1
According to the World Health Organization (WHO)
2
and the U.S. Agency for Healthcare Research and
Quality,
3
intraoperative hypothermia is dened as a
Lohsiriwat et al.
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core temperature less than 36
o
C (96.8
o
F). Intraoperative
hypothermia was associated with poor surgical outcomes
aer major abdominal operations including colorectal
surgery.
4-6
It led to increased surgical bleeding and
requirement of blood transfusion,
7
higher incidence
of cardiac arrhythmia and ischemia,
4
higher rates of
surgical site infection and prolonged hospitalization.
5
Maintaining perioperative normothermia in surgical
patients is therefore an essential part of several surgical
guidelines such as the latest WHO recommendations
for surgical site infection prevention
2
and the Enhanced
Recovery Aer Surgery (ERAS) society recommendations
for perioperative care in elective colorectal surgery.
8,9
Since the adverse eects of intraoperative hypothermia
in colorectal surgery from the published literature were
mainly based on non-ERAS settings,
5,10
strong evidence
supporting this association in patients undergoing
colorectal operations within an ERAS pathway is lacking.
e current study aimed to determine the association
between intraoperative hypothermia and surgical outcomes
following colorectal surgery within an ERAS pathway.
MATERIALS AND METHODS
Patients
A prospective, observational study of adult patients
undergoing elective segmental resection (colectomy
and/or proctectomy) within an ERAS pathway from
March 2011 to October 2015 in the Faculty of Medicine
Siriraj Hospital, ailand was conducted. Patients with
clinical peritonitis or acute colonic obstruction were
excluded. e study was approved by the Institutional
Ethics Committee and written informed consent was
obtained from each patient (Si 014/2013).
Perioperative and operative care
All of the studied patients were operated on and
treated by a board-certied colorectal surgeon (the rst
author) who has applied an ERAS pathway into colorectal
surgery since 2010. ERAS strategies in our institute
were adopted from the ERAS society recommendations
for perioperative care in elective colorectal surgery.
8,9
Some details of our ERAS program have been described
previously.
11-13
Briey, a practice of mechanical bowel
preparation, prophylactic antibiotic regimen, prophylaxis
of postoperative nausea and vomiting, anastomosis creation
with or without stoma formation, analgesic regimen,
early enteral feeding and immediate mobilization was
standardized. However, there was no standardized protocol
of active warming in an operating theater except blood
warmer was used for intraoperative blood transfusion
(if any). Notably, active warming was not standardized
in our ERAS protocol due to the cost and availability of
related equipment. Patients would be discharged from the
hospital if they had no fever, good appetite, satisfactory
gastrointestinal recovery and a good level of ambulation.
All of the patients were scheduled for follow-up at 7-10
days and 30 days aer an operation.
Diagnosis of intraoperative hypothermia
Intraoperative core temperature of the patients was
continuously measured aer induction of anesthesia using
a single esophageal probe which was inserted by a sta
anesthesiologist to the distal half of the esophagus. In this
study patients were classied into a hypothermic group
if their intraoperative core temperature was continuously
below 36
o
C more than 30 minutes. A cut-o period of
half an hour in a hypothermic state (< 36
o
C) was decided
based on a previous study of >50,000 surgical patients
which showed a trend of hypothermia-associated adverse
outcomes from this time point.
14
Data collection
Data including patient characteristics, operative
details, and postoperative outcomes were prospectively
collected. Patient characteristics included age, gender, body
mass index (BMI), American Society of Anesthesiologists
(ASA) grade, and ColoRectal Physiological and Operative
Severity Score for the enUmeration of Mortality and
Morbidity (CR-POSSUM) score.
15
Operative details
included type of operation, operative time, and estimated
blood loss. Overall ERAS protocol compliance of
each case was determined based on the ERAS society
recommendations for perioperative care in elective
colorectal surgery.
8,9
Postoperative outcomes included
postoperative complications (graded I-V according
to the Clavien-Dindo classication system),
16
surgical
site infection (based on the criteria of the U.S. Centers
for Disease Control and Prevention),
17
time to first
bowel movement, time to tolerate normal diet, length
of postoperative stay, death and readmission within 30
days aer the operation.
Statistical analysis
All of the data were prepared and compiled using the
Statistical Package for the Social Sciences program version
18.0 for Windows (SPSS Inc, Chicago, IL). Continuous
variables were expressed as mean ± standard deviation or
median (interquartile; IQR), and were compared using
the Student t-test or Mann-Whitney U test. Categorical
data were expressed as number (percentage) and were
compared using the Pearson Chi-square test or Fisher
exact probability test. Factors inuencing poor surgical
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54
outcomes were analyzed using a univariate analysis. Only
signicant variables from the univariate analysis were
included in a multivariate model of logistic regression,
and the odds ratio with 95% condence intervals (95%CI)
for each variable was determined. A p-value of <0.05 was
considered statistically signicant.
RESULTS
is study included 195 patients: 150 (77%) in
hypothermic group and 45 (23%) in normothermic group.
Maximum, minimum and average intraoperative core
temperature was signicantly lower in the hypothermic
group (Table 1). Patients in the hypothermic group tended
to had a greater volume of intraoperative IV uid (2.4
L vs 1.7 L; p=0.141) and more median blood loss (200
mL vs 100 mL; p=0.074). Patient’s characteristics of each
group are shown in table 1. Factors strongly associated
with intraoperative hypothermia were rectal surgery
(OR=5.15, 95%CI=2.25-11.79; p<0.001), operative time
exceeding 3 hours (OR=3.80, 95%CI=1.82-7.93; p<0.001),
multi-organ resection (OR=3.12, 95%CI=1.04-9.32;
p=0.034) and male gender (OR=2.62, 95%CI=1.30-5.26;
p=0.006).
TABLE 1. Patient’s characteristics and intraoperative parameters.
Hypothermic group Normothermic group P-value
(n=150) (n=45)
Age, year 64.3 ± 13.0 61.6 ± 13.8 0.227
Male 85 (56.7) 15 (33.3) 0.006*
BMI, kg/m
2
23.3 ± 4.5 23.3 ± 4.7 0.968
ASA class ≥ 3 29 (19.3) 8 (17.8) 0.815
CR-POSSUM predictive mortality, % 1.85 (0.98-3.38) 1.37 (0.95-2.58) 0.382
Hematocrit, % 36.7 ± 5.2 35.9 ± 5.7 0.355
Serum albumin, g/dL 3.8 ± 0.6 3.8 ± 0.6 0.368
Cancer surgery 138 (92.0) 39 (86.7) 0.278
Tumor staging ≥ 3 91 (60.7) 26 (57.8) 0.729
Rectal surgery 79 (52.7) 8 (17.8) <0.001*
Multi-organ resection 35 (23.3) 4 (8.9) 0.034*
Laparoscopic surgery 23 (15.3) 11 (24.4) 0.158
Epidural analgesia 48 (32.0) 10 (22.2) 0.208
Total IV morphine consumption, mg/kg 0.11 (0-0.57) 0.10 (0-0.50) 0.976
Core temperature,
o
C
Maximum 36.0 (35.7-36.3) 36.6 (36.4-36.9) <0.001*
Minimum 35.4 (35.0-35.7) 36.1 (36.0-36.4) <0.001*
Average 35.8 (35.4-36.0) 36.5 (36.2-36.6) <0.001*
Duration of surgery, hour 3.7 ± 1.5 3.1 ± 1.4 0.010*
Intravenous uid, L 2.4 ± 1.1 1.7 ± 1.0 0.141
Blood loss, mL 200 (100-400) 100 (50-300) 0.074
Intraoperative blood transfusion, yes 20 (13.3) 4 (8.9) 0.426
Overall ERAS protocol compliance
#
, % 84.4 ± 6.2 85.8 ± 6.5 0.195
* P-value < 0.05
Values are expressed as mean ± standard deviation, median (interquartile range) or number (percentage).
Abbreviations: ASA = American society of Anesthesiologists, BMI = body mass index, CR-POSSUM = ColoRectal Physiological and
Operative Severity Score for the enUmeration of Mortality and Morbidity, ERAS = enhanced recovery aer surgery, IV = intravenous
#Overall compliance of each patient was determined based on the ERAS® society recommendations for perioperative care in elective colorectal
surgery.
Lohsiriwat et al.
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55
Original Article
SMJ
e incidences of postoperative complication and
wound infection were comparable between hypothermic
patients and normothermic patients (23% vs 13%; p=0.17
and 6.0 vs 6.7%; p=0.87, respectively). One patient in
the normothermic group had a 30-day mortality while
the other did not (p=0.12). Hypothermic patients had a
longer time to tolerate normal diet (2.0 days vs 1.3 days;
p=0.023) but a comparable time to rst bowel movement
(2.6 days vs 2.6 days; p=0.84). Hypothermic patients had
a signicant longer hospitalization (5.7 days vs 4.4 days;
p=0.048) (Table 2).
A multivariate analysis adjusted for risk factors
associated with delayed time to tolerate normal food
(> 2 days) showed that intraoperative hypothermia
was an independent predictor (OR=2.9, 95%CI=1.2-
6.9; p=0.014) (Table 3). For predicting prolonged
hospitalization (>5 days), a multivariate analysis showed
that postoperative complication (OR=5.2, 95%CI=2.3-
11.9; p<0.001) and operative time exceeding 3 hours
(OR=3.4, 95%CI=1.8-6.4; p<0.001) were two signicant
risk factors. Intraoperative hypothermia was not associated
with prolonged hospitalization (OR=1.7, 95%CI=0.7-3.9;
p=0.207).
TABLE 2. Postoperative outcomes.
Hypothermic group Normothermic group P-value
(n=150) (n=45)
Overall complication 34 (22.7) 6 (13.3) 0.174
Complication excluding grade I
#
19 (12.7) 3 (6.7) 0.265
Wound infection 9 (6.0) 3 (6.7) 0.870
30-day death 0 1 (2.2) 0.231
30-day readmission 5 (3.3) 3 (6.7) 0.389
Time to tolerate normal diet, days 2.0 ± 2.0 1.3 ± 1.3 0.023*
Time to rst bowel movement, days 2.6 ± 1.1 2.6 ± 1.1 0.838
Length of hospitalization, days 5.7 ± 4.2 4.4 ± 2.6 0.048*
*P-value < 0.05
Values are expressed as mean ± standard deviation or number (percentage).
#According to the Clavien-Dindo classication of surgical complications
TABLE 3. Multivariate analysis of factors potentially associated with delayed time to tolerate normal diet (>2 days).
Oddsratio 95%condentialinterval P-value
Intraoperative hypothermia 2.88 1.20-6.90 0.014*
Postoperative complication 1.99 0.97-4.09 0.059
Operative time exceeding 3 hours 1.81 0.97-3.38 0.059
Rectal surgery 1.58 0.86-2.92 0.142
Open surgery 1.32 0.61-2.89 0.482
Hypoalbuminemia 1.04 0.50-2.19 0.912
No epidural analgesia 0.84 0.44-1.64 0.621
Multi-organ resection 0.75 0.34-1.66 0.483
*P-value < 0.05
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56
DISCUSSION
is study of 195 patients showed that intraoperative
hypothermia was an independent risk factor for prolonged
time to tolerate normal food in patients undergoing elective
colorectal surgery. However, there was no association
between intraoperative and time to rst bowel movement,
overall complication, surgical site infection or length of
hospitalization. Rectal surgery, operative time exceeding 3
hours, multi-organ resection and male patients perceived
greater risks of intraoperative hypothermia.
is study showed that intraoperative hypothermia
was associated with delayed time to tolerate normal diet.
Hypothermia-associated prolonged GI recovery could
be explained by several possible mechanisms. First, the
sympathetic nervous system is stimulated during the
period of hypothermia to generate heat production
and prevent further heat loss
18
. Neurotransmitters of
the sympathetic nervous system such as adrenaline and
noradrenaline are known to decrease GI motility and
reduce luminal secretion.
19,20
Sympathetic stimulation also
led to an inhibition of the vagus nerve-mediated gastric
contractions
21
and decreased food appetite.
22
Second, the
abnormal activities of the sympathetic nervous system
in the GI tract may cause gut inammation and motility
disorders.
23
ird, in animal studies cold temperature
diminished spontaneous movements of small bowel
and depressed acetylcholine-induced contraction thus
indicating that the tonic and phasic component of small
bowel contraction are sensitive to cold temperature.
24
Although intraoperative hypothermia was an independent
predictor for delayed time to tolerate solid food, it did not
aect time to rst bowel movement or time to discharge
patients.
Within an ERAS setting this study failed to demonstrate
a correlation between intraoperative hypothermia and
postoperative complications including wound infection.
ese ndings are similar to those reported in several
large and recent studies examining an association between
perioperative hypothermia and surgical site infection
following colorectal surgery.
10,25,26
For example, Baucom and
her colleagues showed that, regardless of how hypothermia
was dened, intraoperative temperature did not predict
infectious complications aer laparoscopic and open
colorectal operations.
10
Linking to the American College
of Surgeons National Surgical Quality Improvement
Program, Melton et al also did not nd any correlation
between intraoperative hypothermia and 30-day surgical
site infection in 1008 colorectal procedures.
26
Our ndings were in contrast to the 1996 landmark
study by Kurz et al which was a randomized prospective
trial of routine care versus additional intraoperative
warming in 200 patients undergoing open colorectal
resection in non-ERAS setting.
5
Kurz et al reported that
patients with hypothermia had three times higher rates
of wound infection (19% vs 6%) and 2.6-days longer
hospitalization compared with normothermic patients.
Although hypothermic patients in our study had 1.3-
days longer hospitalization than the other group, in a
multivariate analysis the prolonged hospitalization was a
result of postoperative complications – not intraoperative
hypothermia. Notably, in our study the rates of wound
infection in both studied groups were comparable (6% in
hypothermic patients and 6.7% in normothermic patients)
and almost identical to those with active warming in the
study of Kurz et al. It is conceivable that the detrimental
eects of ‘mild’ intraoperative hypothermia on surgical
site infection may be negligible in an ERAS setting.
Within an ERAS pathway, the implementation of bundled
interventions including appropriate administration of
prophylactic antibiotics and better glycemic control
signicantly decreased the rates of surgical site infection
aer colorectal operation to 4-7%.
27
e incidence of intraoperative hypothermia in
our study was high (77%). is may be explained by
the fact that active warming protocol and standardized
maneuvers for preventing hypothermia are lacking in
our institute even an ERAS pathway has been applied for
several years.
11-13
In some institutes, where the routine
use of body-warming devices and other eorts to prevent
and manage perioperative hypothermia, the incidence of
intraoperative hypothermia may be as low as 7%.
28
Forced
air warming system appeared to be the most ecient
in maintaining perioperative normothermia compared
with reective blanket and warmed cotton blanket.
29
Warming of large amounts of intravenous uid, blood
and inspired air is also commonly used in the theater
to preventing hypothermia in developed countries.
26,30
However, the rate of active warming of patients during
an operation is low in resource-poor countries including
ailand and other Asian countries.
31-33
Our data indicated that rectal surgery, operative time
exceeding 3 hours, multi-organ resection and male gender
were signicant predictors for intraoperative hypothermia.
Several risk factors for intraoperative hypothermia have
been identied in the literature including high ASA physical
status, major surgery, operative time exceeding 2-3 hours,
use of combined epidural and general anesthesia and
intravenous administration of un-warmed uid or blood
components.
33
Meanwhile, active warming, overweight,
high baseline core temperature before anesthesia and
high ambient temperature were signicant protective
factors for intraoperative hypothermia.
32
Lohsiriwat et al.
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57
Original Article
SMJ
Our study benets from the use of a single-center
database of ERAS pathway in colorectal operations.
Notably, the patients in this registry were taken care of by
single surgeon’s team with good adherence to the ERAS
protocol. However, there are several limitations of this
observational study. First, the sample size was relatively
small. Potential negative impact of hypothermia on
surgical outcomes reported in non-ERAS setting, such as
prolonged hospitalization,
5
may be not clearly evident in
this study due to the low sample size. Second, the eect of
intraoperative hypothermia was evaluated only in patients
undergoing colorectal surgery – mainly open surgery for
colorectal cancer, making it dicult to extrapolate our
results to patients undergoing other operations. ird,
it could be argued that active warming is currently the
accepted standard of care and laparoscopic surgery has
become a common approach with a less incidence or less
degree of intraoperative hypothermia. We acknowledge
that it is true in developed countries but maybe not
in developing and underdeveloped regions
31,33
– thus
making this study a great opportunity to re-evaluate
the eect of intraoperative hypothermia in the current
surgical practice. Furthermore, it would be interesting
to examine in the future whether cost savings from the
omission of active warming is o-set by additional costs
to provide care in the postoperative period. Finally, our
ndings were analyzed based on a denition of ‘mild’
intraoperative hypothermia. Whether moderate or severe
hypothermia will adversely aect surgical outcomes
under an ERAS pathway needs to be examined.
In conclusion, despite these limitations, our data
indicated that intraoperative hypothermia prolonged
time to resume normal food aer colorectal surgery
within an ERAS setting but it did not adversely aect
the return of bowel function, surgical site infection,
postoperative complications and length of hospitalization.
ese ndings suggest that the detrimental eects of
‘mild’ intraoperative hypothermia on surgical outcomes
may be minimal in an ERAS setting.
ACKNOWLEDGMENTS
The author would like to thank Mr. Suthipol
Udompunthurak from Clinical Epidemiology Unit,
the Oce for Research and Development, Faculty of
Medicine Siriraj Hospital, for his kind assistance with
statistical analysis.
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15. Tekkis PP, Prytherch DR, Kocher HM, Senapati A, Poloniecki
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Lohsiriwat et al.
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Original Article
SMJ
Saifon Chawanpaiboon, M.D., Vitaya Titapant, M.D.
Department of Obstetrics & Gynaecology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok 10700, ailand.
A Randomized Controlled Trial of the Correlation
between Iodine Supplementation in Pregnancy and
Maternal Urine Iodine and Neonatal Thyroid
Stimulating Hormone Levels
Corresponding author: Saifon Chawanpaiboon
E-mail: saifon.cha@mahidol.ac.th
Received 31 July 2018 Revised 22 August 2018 Accepted 18 October 2018
ORCID ID: 0000-0002-3207-6187
http://dx.doi.org/10.33192/Smj.2019.10
ABSTRACT
Objective: To establish the correlation of maternal urine iodine and neonatal thyroid stimulating hormone (TSH)
in iodine supplemented and non-iodine supplemented pregnant women.
Materials and Methods: e study was a prospective, randomized, controlled trial, which was conducted at the
antenatal care unit, labor ward, and neonatal unit of Siriraj Hospital, Mahidol University, Bangkok, ailand. In
all, 224 pregnant women were recruited during 1 October 2015 and 31 July 2017. ey were randomized into 2
groups: an iodine and a non-iodine supplemented group. One woman in the control group le the study as she
had delivery at another hospital.
Results: ere were no statistically signicant dierences in the demographic data, original area of domicile, and
adverse neonatal outcomes (including preterm labor and low birth weight) of the patients in the two groups. e
maternal urinary iodine levels were 84.14 ± 61.85 and 58.41 ± 41.36 microgram/L, and the median values of the
neonatal TSH levels were 3.7 ± 1.87 and 4.4 ± 1.99 mIU/ml, in the iodine and non-iodine supplemented groups,
respectively. e dierences in both values were statistically signicant (p-value < 0.05).
Conclusion: is study determined that there were statistically signicant dierences in the maternal urinary iodine
levels and the median values of the neonatal TSH levels of the iodine and non-iodine replacement groups of pregnant
women. Even though there were no clinically signicant dierences and none of the newborns was diagnosed with
hypothyroidism, iodine supplementation in all pregnant women should be considered. A larger prospective, RCT
trial would conrm the benets of a strategy of routinely administering iodine to pregnant women at Siriraj Hospital.
Keywords: Iodine supplementation; maternal urine iodine; neonatal TSH; hypothyroidism (Siriraj Med J 2019;71:
59-65)
INTRODUCTION
Iodine is an essential substance for fetal brain
development; cell metabolism; cell growth; and the
myocardial, hepatic, and muscle functions. yroid
hormone production requires iodine as the major
substance. Iodine deciency results in impaired thyroid
hormone synthesis and/or thyroid enlargement (goiter),
hypothyroidism, cretinism, a decreased fertility rate,
increased infant mortality, and mental retardation
1
, as well
as miscarriage and preterm labor in pregnant women.
2
As
the development of the fetal thyroid gland and hormone
production are delayed during gestation, the fetus is
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60
totally dependent during early pregnancy on maternal
thyroxine for normal brain development.
3
Maternal
dietary supplementation of iodine during pregnancy
is also benecial. Irreversible fetal brain damage can
result from inadequate iodine supplementation during
pregnancy, which may lead to inadequate production
of thyroid hormones and hypothyroidism in pregnant
women.
3
Iodine supplementation before and during
pregnancy can prevent cretinism and improve the cognitive
function of the general population.
1
e growing requirement for iodine in pregnancy
arises from the progressive increase in maternal thyroxine
(T4) production needed to maintain maternal euthyroidism
and to transfer thyroid hormone to the fetus during
the rst trimester, before the fetal thyroid begins to
function. Other reasons are an iodine transfer to the fetus,
particularly in later gestation, and an increase in renal
iodine clearance.
4
e US Institute of Medicine
5
and the
World Health Organization (WHO)
6
recommendations
for iodine intake during pregnancy are 220 and 250
micrograms/day, respectively. From WHO global estimates
of iodine status, more than half of the children with
iodine deciency came from Southeast Asia and Africa.
7
ailand has an iodine deciency, even though the
south and southeast parts of the country have substantial
coastal areas. In 2013, a national survey found that iodine
deciency was a major problem among pregnant ai
women. Nearly half of all the women suering from
iodine deciency were reported to be located in the north
and northeast of ailand, neither of which are near the
coast.
8
ailand’s capital city, Bangkok, has plenty of sea
food available for consumption. Almost all ai food in
that city is cooked with the ingredients of sh sauce and
iodized salt, which may result in adequate iodine intake
by pregnant ai women; hence, iodine supplementation
may be unnecessary for pregnant women in Bangkok.
Prior to the present study, no research had been
conducted of the degree of correlation between iodine
supplementation in pregnancy and the maternal urine
iodine and neonatal TSH levels of patients at Siriraj
Hospital. is research was carried out to determine the
need for iodine-containing medicated supplementation
by pregnant ai women. We hypothesized that iodine
supplementation would still be necessary despite pregnant
ai women having access to adequate seafood nutrition.
MATERIALS AND METHODS
e study was approved by the Ethics Committee of
the Siriraj Institutional Review Board (Si 524/2014). is
prospective, randomized, controlled trial was conducted
at the antenatal clinic and labor ward, Faculty of Medicine
Siriraj Hospital, Mahidol University, from October 2015
to July 2017. Using nQuery Advisor (Statistical Solutions
Ltd., Cork, Ireland), the calculation of the sample size
was based on the ndings of a study from Denmark
9
,
which reported that the median values of the neonatal
TSH levels in pregnant women with and without iodine
supplementation were 9 mU/l and 7.07 mU/l, respectively.
However, the standard deviation (SD) employed in the
present study’s calculations was 4 times greater than that
used in the Danish study to account for the variations
in the populations of many parts of ailand. A 10%
follow-up loss was also factored in. e nal sample size
was determined to be 112 cases for each group.
Included in the study were all pregnant women who
were older than 18 years and who had a singleton fetus at a
gestational age of less than 18 weeks on the day the women
commenced participation. e exclusion criteria were
patients who had any of the following: a contraindication
to the use of iodine, a previous administration of other
iodine-containing drugs, a multifetal pregnancy, a pregnancy
with a fetal anomaly, or a pregnancy with an abnormal
thyroid function (hyperthyroidism or hypothyroidism).
e total of 224 pregnant women were recruited and
divided into 2 groups of 112 by block randomization (block
sizes of ten). One group received an iodine-containing
ferrous tablet, and the other a no iodine-containing
ferrous tablet. e patient’s demographic data were
obtained; the data items comprised age, pre-pregnant
body weight, height, occupation, income, area of domicile,
socioeconomic status, parity, antenatal care history, and
medications received during pregnancy.
All of the pregnant women were given the standard
care aorded to other patients, such as gestational diabetes
mellitus and thalassemia screening, ultrasonography, and
other indicated fetal surveillances. In the case of anemic
patients (dened as a hematocrit level under 33%), an
iron supplement (FeSO4 tablets) was prescribed.
e primary objective was to nd any correlations
between the neonatal TSH levels at 48-hours of life in
the iodine supplemented and non-iodine supplemented
groups. e reference cut-o value of an abnormally high
TSH level used by the study was 12 mIU/L. is value
was based on the laboratory reference range provided
by the Genetics Division, Pediatrics Department, Siriraj
Hospital.
All descriptive data were analyzed by descriptive
statistics, and an unpaired t-test or Mann–Whitney U
test was used to analyze the correlation of the data and
the neonatal TSH levels. e data was deemed to have
statistical signicance at a p-value of less than 0.05.
Chawanpaiboon et al.
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RESULTS
Of the total of 224 pregnant women initially recruited,
1 woman in the control group le the study as she had
delivery at another hospital.
e demographic data and neonatal outcomes are
presented in Tables 1 and 2. e patients’ data included
the mean age, parity, body weight, height, body mass
index (BMI), gestational age at the rst antenatal care
unit visit and later at the start of medication, occupation,
education level, and monthly income. ere were no
statistically signicant dierences between the data for
the two groups (p-value < 0.05).
As to the neonatal outcomes, the preterm birth rates
of the patients in the iodine and non-iodine supplemented
groups were 8.2% and 7.1%, and the rates of low birth
weights were 6.1% and 10.1%, respectively. No statistically
signicant dierences were detected in those gures.
e mean gestational ages at delivery were 38.1 and
38.3 weeks, and the mean neonatal birth weights were
3,061.8 + 474.7 and 3,075.9 + 407.4 grams, respectively,
again with no statistically signicant dierences (Table 3).
e mean urinary iodine level of the patients in the
iodine supplement group was 84.14 ± 61.85 microgram/L,
which was higher than that of the patients in the non-
iodine supplemented group (58.41 ± 41.36 micrograms/L;
Table 5). However, the median neonatal TSH level of the
patients in the iodine supplemented group was 3.7 ± 1.87
mIU/L, which was lower than the gure of 34.4 ± 1.99
mIU/L for the patients in the non-iodine supplemented
group. ere were statistically signicant dierences in
the results for the two groups (p-values 0.001 and 0.01,
respectively; Table 6).
DISCUSSION
e most authoritative guidelines on how to assess
iodine nutrition in a population were published in 2007
by the WHO, the United Nations Children’s Fund
(UNICEF), and the International Council for Control
of Iodine Deficiency Disorders (ICCIDD, currently
named the Iodine Global Network).
10
Iodine deciency
is the most common micronutrient deciency in the
world, especially in Asia.
7
ailand also has an iodine
deciency even though plenty of seafood is available in
most areas of the country. e WHO and the ICCIDD
have promoted the usage of iodized table salt to alleviate
endemic cretinism in many parts of the world.
10
Despite
iodine deciency aecting both the mother and the child,
iodine supplementation during pregnancy tends to be
of low concern among physicians in ailand. Pregnant
women require a 50% increase in their iodine intake to
ensure there is sucient available for thyroid hormone
production by fetuses.
11
However, inadequate iodine
supplementation may result in an iodine deciency for
both mothers and fetuses; alternatively, the mother may
achieve euthyroidism, yet the fetus becomes hypothyroid.
In 2013, the annual statistical report of the Pediatric
Genetic Division of the Pediatrics Department, Faculty
of Medicine Siriraj Hospital, reported a case of a neonate
that had an abnormally high TSH level of 0.17%, yet only
0.02% of the neonates in the group of neonates with
abnormally high TSH levels had been diagnosed with
hypothyroidism.
12
is proportion seems to be small and
may not be representative of the extent of hypothyroidism
among ai children generally. However, the current study
found that the urine iodine and neonatal TSH levels for
TABLE 1. Demographic data of the study groups.
Demographic data Iodine supplemented Non-iodine supplemented P-value
+
group (N = 112) group (N = 111)
Mean ± SD Mean ± SD
Age 29.9 ± 5.8 29.5 ± 5.8 0.64
Body weight 54.8 ± 10.5 54.9 ± 11.1 0.92
Height 157.9 ± 6.4 158.7 ± 5.5 0.39
BMI 21.9 ± 3.9 21.8 ± 4.0 0.76
GA at 1st ANC unit visit 10.3 ± 3.2 11.0 ± 3.7 0.17
GA at start of medication 14.9 ± 2.1 14.9 ± 2.5 0.15
BMI, body mass index; +, t-test; GA, gestational age; ANC, antenatal care
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TABLE 2. Demographic data of the study groups.
Demographic data Iodine supplemented Non-iodine supplemented P-value
+
group (N = 112) group (N = 111)
N (%) N (%)
Parity 0.33
1 39 (34.8) 52 (46.8)
2 47 (42.0) 34 (30.6)
3 21 (18.8) 16 (14.4)
4 5 (4.5) 7 (6.3)
5 0 (0) 2 (1.8)
Occupation 0.39
Housewife 16 (14.3) 22 (19.8)
Farmer 0 (0) 1 (0.9)
Governmentofcer 4(3.6) 8(7.2)
Stateenterpriseofcer 3(2.7) 1(0.9)
Laborer 68 (60.7) 62 (55.8)
Merchant 16 (14.3) 16 (14.4)
Other (student, business owner, 5 (4.5) 1 (0.9)
unemployed)
Income (Baht/mo) 0.57
< 10,000 13 (11.6) 9 (8.1)
10,000–29,999 66 (58.9) 79 (71.1)
30,000–49,999 21 (18.8) 15 (13.5)
> 50,000 12 (10.7) 8 (7.2)
Education 0.64
Primary school 1 (0.9) 0 (0)
Secondary school 43 (38.4) 39 (38.6)
Bachelor and higher degree 68 (60.7) 72 (64.9)
+
, Chi-square test
TABLE 3. Neonatal outcomes.
Neonatal outcome Iodine supplemented Non-iodine supplemented P-value
+
group (N = 112) group (N = 111)
Mean (SD) Mean (SD)
GA at delivery 38.1 ± 1.7 38.3 ± 1.9 0.97
Neonatal birth weight 3,061.8 ± 474.7 3,075.9 ± 407.4 0.58
GA, gestational age;
+
, Chi-square test
Chawanpaiboon et al.
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TABLE 4. Incidences of preterm births and low birth weights.
Iodine supplemented Non-iodine supplemented P-value
+
group (N = 112) group (N = 111)
N (%) N (%)
Preterm birth (GA < 37 wk) 11 (9.8) 10 (9.0) 0.9
Extremely preterm(< 28 wk) 0 (0) 0 (0)
Very preterm (28– < 32 wk) 2 (1.7) 1 (0.1)
Moderate to late preterm (32– < 37 wk) 7 (6.2) 6 (5.4)
LBW (birth weight < 2,500 g) 8 (7.1) 10 (9.0) 0.36
Extremely LBW (< 1,000 g) 0 (0) 0 (0)
Very low birth weight (1,000– < 1,500 g) 2 (1.8) 0 (0)
Low birth weight (1,500– < 2,500 g) 6 (5.3) 10 (9.0)
GA, gestational age;
+
, Chi-square test
TABLE 5. Urinary iodine levels of the two groups.
Groups N Mean of urinary iodine P-value
+
(range) microgram/L
Iodine supplemented group 112 84.14 ± 61.85 0.001
(9.40 – 437.00)
Non-iodine supplemented group 111 58.41 ± 41.36
(3.01–215.60)
+
, Chi-square test
TABLE 6. Median of neonatal TSH levels of the two groups.
Groups N Median of neonatal P-value
+
TSH (range) mIU/ml
Iodine supplemented group 112 3.7 ± 1.87 0.01
(0.66–11.30)
Non-iodine supplemented group 111 4.4 ± 1.99
(1.31–10.60)
+
, Chi-square test
those mothers with iodine supplementation were higher
and lower, respectively, than the corresponding gures
for the mothers without iodine supplementation. It has
been reported that children born from mothers with an
iodine deciency may lose up to 13.5 IQ points.
13
e
International Child Development Steering Group has
identied that iodine deciency is one of the four, key,
global health factors for impaired child development
that have the most urgent need for intervention.
14
As about 90% of absorbed iodine is excreted in
urine, the median urinary iodine concentration (MUIC)
is the best indicator of iodine intake.
15
e MUIC in the
general population should be between 100 and 199 mcg/L,
while in pregnant women, it should be in the range of
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64
150 to 249 mcg/L. In our study, the MUIC in the group
of pregnant women with iodine supplementation was
only 84.14 ± 61.85 (9.4–437) mcg/L, which was lower
than the standard requirement.
e neonatal TSH level is a biological indicator in
national congenital hypothyroid screening programs.
e guidelines of WHO/ICCIDD/UNICEF state that a <
3% frequency of TSH values > 5 mIU/L (in whole blood
spots) indicates iodine suciency in a population. is
numerical value was measured from 72 hour–old neonates
who were born in iodine sucient areas. Our study found
that the median value of the TSH from the neonates of
the mothers receiving iodine supplementation was 3.7
± 1.87 (0.66–11.3) mIU/ml, which was lower than those
without iodine supplementation. However, Smyth
16
and
Li
17
suggested that using neonatal TSH levels may not
be a reliable method for indicating an iodine deciency
in newborns because of the discrepancy between the
MUIC and neonatal TSH levels.
e results of our study indicated that even in
pregnant women receiving iodine supplementation, the
MUIC and neonatal TSH levels were still abnormal. e
main consensus in the WHO/UNICEF/ICCIDD guidance
on achieving an adequate iodine intake in the general
population as well as in pregnant women is that salt
iodization is the key strategy.
10
Iodine supplementation
during pregnancy is important, but encouraging the
usage of iodized salt should be the primary strategy for
all pregnant women. e consensus reached by WHO/
UNICEF/ICCIDD was that pregnant women should not
be advised to take iodine-containing supplements if the
general population they come from is iodine sucient,
indicated by that population having a median UIC ≥
100 µg/l for at least 2 years.
18
In ailand’s case, the
country generally has an iodine deciency; therefore,
household iodized-salt usage and supplementation with
iodine-containing iron during pregnancy should prevent
iodine deciency in newborns.
Encouraging the long-term use of household iodized
salt is the most eective strategy for eradicating iodine
deficiency.
10
Even though the Thai National Iodine
Deciency Disorder Control Project has promoted the
regular use of household iodized salt since 1989, an
iodine deciency among pregnant ai women was still
reported in national surveys conducted during the years
2000–2006.
19
e dual promotion of household iodized
salt usage and of the prescribing of iodine-containing
iron supplementation during pregnancy has been in force
since October 2010. A previous study from ailand
reported an improvement in the iodine nutrition of
pregnant women aer the implementation of those health
policies.
20
However, our study showed that although the
iodine-containing iron supplementation aected the
MUIC and neonatal TSH levels, the MUIC level was
still lower than that recommended by WHO/UNICEF/
ICCIDD. Extensive repromotion of household iodized
salt should be considered in order to improve the iodine
nutrition of pregnant women.
e limitation of this study was that the baseline
iodine status before the iodine supplementation was
not measured. erefore, some of the pregnant women
may have had a severe iodine deciency status prior to
attending the study, which would have aected the MUIC
and neonatal TSH levels. Moreover, it was not possible
to control the daily iodine level of the participants’ diets,
which could have aected the iodine status of some of
the pregnant women.
Many studies have supported the view that iodine
supplementation during pregnancy can improve iodine
status and neonatal TSH levels.
20–22
Providing iodine
supplementation to all pregnant women is still benecial
to neonatal brain development.
e strength of our study was that it was a randomized
controlled trial. e sequence generation and allocation
were well designed, which was evidenced by there being
no signicant dierences in the demographic data of the
2 groups. Nevertheless, the study had some limitations.
ese were some foreign babies missed the screening
of the TSH levels due to their parents being unable to
aord the related costs, an incompleteness of iodine
supplementation, a lack of assessment of the medication
adherence by both groups, and the absence of basis data
relating to iodine status before participation in the study.
In order to obtain more precise data on the benets
of iodine supplementation during antenatal care, a larger
prospective RCT should be performed. Moreover, the
baseline iodine status should be determined before
commencing the iodine supplementation.
CONCLUSION
ere were statistically signicant dierences in the
mean levels of the urinary iodine and the median values
of the neonatal TSH levels of the iodine and non-iodine
supplement groups of pregnant women. Even in areas with
plentiful supplies of iodized food, pregnant women still
had an iodine deciency. us, iodine supplementation
for all pregnant women should be encouraged, even if
only to ensure proper fetal brain development.
What is already known on this topic
From previous studies, iodine has been established
as being essential for fetal brain and thyroid development,
Chawanpaiboon et al.
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65
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and it is recommended for all pregnant women. However,
the administration of iodine supplementation for pregnant
women is not routinely applied at all centers in ailand,
especially at Siriraj Hospital, which is a tertiary center.
e general belief among health professionals that there
is sucient iodine intake during pregnancy has led to a
lack of concern about the need for iodine supplementation
during pregnancy at Siriraj Hospital. e objective of
this study was to ascertain the TSH levels of neonates of
iodine- and non-iodine-supplemented mothers during
pregnancy.
What this study adds
e study found a statistically signicant dierence
in the median values of the neonatal TSH levels of the
iodine and non-iodine supplemented groups. Even though
there were no clinical signs of hypothyroidism in the
neonates, iodine is still benecial for brain development.
is study indicated that iodine supplementation aected
the TSH levels of neonates and should therefore be
adopted for all pregnant women in ailand, especially
at Siriraj Hospital. A larger study is needed to conrm
the benets of widespread iodine supplementation.
ACKNOWLEDGMENTS
e authors thank Professor Prasit Watanapa, Dean
of the Faculty of Medicine, Siriraj Hospital, for his support
of research in residency training, and Nattacha Palawat
for her administrative support. is research project
was supported by Faculty of Medicine, Siriraj Hospital,
Mahidol University (Grant Number [IO] R015831068).
Potential conicts of interest
We have no potential conicts of interest.
REFERENCES
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disorders. Lancet. 2008;372(9645):1251-62.
2. Zimmermann MB. Iodine deciency. Endocr Rev. 2009;30(4):376–
408.
3. Public Health Committee of the American yroid A, Becker
DV, Braverman LE, Delange F, Dunn JT, Franklyn JA, et al.
Iodine supplementation for pregnancy and lactation—United
States and Canada: recommendations of the American yroid
Association. yroid. 2006;16(10):949-51.
4. Glinoer D. e regulation of thyroid function during normal
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Nickel, Silicon, Vanadium, and Zinc. Washington (DC)2001.
6. Assessment of Iodine Deciency Disorders and Monitoring
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7. Andersson M, Karumbunathan V, Zimmermann MB. Global
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9. Nohr SB, Laurberg P. Opposite variations in maternal and
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Pollitt E, et al. Child development: risk factors for adverse
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http://whqlibdoc.who.int/publications/2007/9789241595827_eng.
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16. Burns R, Mayne PD, O’Herlihy C, Smith DF, Higgins M, Staines
A, et al. Can neonatal TSH screening reect trends in population
iodine intake? yroid. 2008;18(8):883–8.
17. Li M, Eastman CJ. Neonatal TSH screening: is it a sensitive
and reliable tool for monitoring iodine status in populations?
Best Pract Res Clin Endocrinol Metab. 2010;24(1):63-75.
18. Secretariat WHO, Andersson M, de Benoist B, Delange F,
Zupan J. Prevention and control of iodine deciency in pregnant
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P, Panburana P, Chailurkit LO, Khlairit P, et al. Eects of
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Anticha Siritongtaworn, M.D., Puriwat To-adithep, M.D., Onuma Chaiwat, M.D.
Department of Anesthesiology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok 10700, ailand.
The Implementation of a Red Blood Cell Transfusion
Guideline in Critically Ill Surgical Patients at Siriraj
Hospital
Corresponding author: Onuma Chaiwat
E-mail: onuma.cha@mahidol.ac.th
Received 17 August 2017 Revised 17 October 2017 Accepted 8 December 2017
ORCID ID: 0000-0003-4464-9226
http://dx.doi.org/10.33192/Smj.2019.11
ABSTRACT
Objective: To compare the RBC transfusion rate and clinical outcomes before and aer the implementation of a
transfusion guideline and to determine the adherence rate to the guideline.
Methods: is is a prospective experimental study in adult patients (≥18 years) admitted to surgical intensive care
units. Aer developing and implementing a transfusion guideline, the data including patients’ characteristics,
transfusion and outcomes were collected prospectively (post-educational group). Data in pre-educational group
was retrieved retrospectively from medical records.
Results: ere were 197 patients in pre-educational group and 188 patients in post-educational group. e incidence
of RBC transfusion signicantly decreased signicantly in post-educational group (37.2% vs. 48.7%, p = 0.02). e
hemoglobin threshold for RBC transfusion was signicantly lower in post-education group (8.16±1.43 vs. 8.97±1.57,
p=0.001). e 28-day mortality rate was signicantly decreased aer the implementation of the guideline. (3.2 %
vs. 11.2 %, p = 0.007). e adherence rate to transfusion guideline was reported as 47.1%.
Conclusion: e incidence of RBC transfusion was lower aer the implementation of a transfusion guideline.
Keywords: Red Blood cell; transfusion; surgical patients; intensive care; guidelines (Siriraj Med J 2019;71: 66-73)
BACKGROUND
Anemia has been associated with poor clinical outcomes
including higher mortality in patients who underwent
non-cardiac surgery.
1
Red blood cell transfusions (RBCs)
is one of the available methods that is frequently used to
treat anemia or hemorrhage in order to improve oxygen
delivery to tissues. Nevertheless, it is well established
that RBC transfusion is an independent predictor of
death, nosocomial infection, multi-organ dysfunction
syndrome (MODS) and acute respiratory distress syndrome
(ARDS) in critically ill patients.
2
Transfusion related
immunomodulation (TRIM) is accounted to be the
underlying cause.
3
However, many studies from diverse locations in
North America and Western Europe have demonstrated
that 30-40% of the patients admitted to intensive care
unit (ICU) still received RBC transfusion.
4-6
In addition,
it was increasing up to 73% in patients with an ICU-stay
longer than 7 days.
4
e pre-transfusion hemoglobin
(Hb) was reported around 8.5 g/dl. Surgical patients have
shown to receive more blood transfusions than medical
patients, particularly in those with emergency surgery.
4
A retrospective study from Siriraj Hospital in critically ill
surgical patients who had been on mechanical ventilation
for greater than 24 hours reported an incidence of RBC
transfusion was as high as 83 % and the mean RBC
Uataya et al.
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SMJ
transfusion threshold was at Hb level of 8.7±1.2 g/dL.
7
e morbidity and mortality were higher in transfused
patients.
7
Although recent data suggested that restrictive
transfusion strategy with the a Hb threshold of 7.0 g/
dl was well tolerated in general critically ill patients
8,9
,
trauma patients
10
, sepsis patients
11,12
and in cardiac surgery
patients
13,14
concerns have been raised regarding the
actual practices and the remaining high incidence of
RBC transfusion.
In addition, there has been lack of high quality data
regarding the optimal Hb threshold in certain critically
ill populations such as those with preexisting coronary
disease, cerebrovascular disease and renal failure. e
implementation of a guideline with species indication
for transfusion might help to reduce the unnecessary
transfusion. Although a number of previous guidelines
12,15,25
have been published, the adherence to the guideline might
be problematic. Educational program including training
course and posters related to the specic guideline for
transfusion probably improves the adherence rate to the
guideline. As a result, lower RBCs transfusion rate and
better clinical outcomes are anticipated. is study aims
to compare the RBC transfusion rate before and aer
the implementation of a guideline and the adherence
rate to the guideline in critically ill surgical patients.
MATERIALS AND METHODS
e study was approved by Siriraj Institutional
Review Board (Si 602/2013) with the waiver of informed
consent. It was a prospective experimental study. All
surgical patients admitted to SICU with age of equal
to or more than 18 years during the study period were
included. Patients with brain death or imminent death
required withholding or withdrawing the treatment or
had demonstrated an active blood loss (dened as blood
loss of more than 30% of blood volume) were excluded.
e study consisted of 5 periods
• 1 month for developing a transfusion guideline and
distributing a guideline to ICU physicians and surgeons
by setting the meeting and ocial letters to the head of
department
• Aer an approval from SIRB, 4 months pre-
guideline data collection (April to August 2011) from the
previous medical records were obtained (pre-educational
group)
• 1 month (February 2014) for providing educational
programs to the anesthesia attending sta who were the
same group in both pre- and post-educational period
and anesthesia resident trainees who were working in
ICU during the study period. Programs included formal
lecture at the beginning, poster presentation, reminder
card and reminder about transfusion guideline once a
month
• 4 months prospective observational period with
data collection aer providing educational programs
between March and July 2014 (post-educational group)
• 2 months of follow up period
Data collection were 1) patient proles including age,
gender, weight, height, primary diagnosis, co-morbid
disease (diabetes mellitus, hypertension, cardiovascular
disease, chronic lung disease, chronic kidney disease,
hematologic disease), smoking, alcohol drinking, Acute
Physiology and Chronic Health Evaluation II (APACHE
II) score, type and duration of surgery, type and duration
of anesthesia and the presence of sepsis on ICU admission;
2) transfusion data (post-educational group) including
Hb/hematocrit (Hct) level before RBC transfusion. In
case, the patients were transfused more than one time,
we collected the transfusion information at the rst
transfusion), indication for RBC transfusion and the
number, type and storage time of the transfused RBC;
3) clinical outcomes including nosocomial infection,
cardiovascular events, acute lung injury (ALI)/ARDS,
and MODS, ventilator days, ICU and hospital length of
stay and 28-day mortality rate
Guideline for transfusion of RBC in critically ill surgical
patients, Siriraj Hospital
15, 16
Preface
1. World Health Organization (WHO) defines
anemia as hemoglobin concentration (cHb) < 13.0 g/dl
in men and < 12.0 g/dl in women.
2. Except dramatic emergency events with
exsanguinating patients, transfusion of RBC should be
generally performed as single-unit transfusion, which
means one unit a time; the next unit, if necessary is given
aer actual Hb recording.
3. Any varyiation from the standards described
below should be explained by the decision maker and
documented in the patients le.
Indications for red cell transfusion (Fig 1)
Patients with prolonged hemorrhagic shock or
acute hemorrhage (> 30% blood volume) that is dicult
to estimate and not manageable by colloid/crystalloid
infusion, and signs of oxygen deciency, such as drop
in central venous oxygen saturation (ScvO
), increase in
arterio-venous dierences of oxygen (AVDO
2
), elevated
plasma lactate level and increase in base excess (BE).
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Fig 1. Transfusion Guidelines in surgical ICUs, Siriraj Hospital.
Additional indications
A. Hemoglobin concentration (cHb) < 7 g/dl in
normovolemic anemia without pathologic symptoms
related to anemia.
A. 1 Symptoms of anemia
I. Chest pain (deemed to be cardiac in origin)
II. Congestive heart failure
III. Otherwise unexplainable tachycardia
Situation has to be reassessed aer each RBC unit given.
B. Hemoglobin concentration (cHb) < 8 g/dl in
normovolemic anemia with history of:
I. Ischemic heart disease
II. Electrocardiographic evidence of previous
myocardial infarction
III. Presence of congestive heart failure
IV. Stroke or transient ischemic attack
C. Hemoglobin concentration (cHb) < 10 g/dl in
patients with acute myocardial infarction or unstable
angina.
Statistical analysis
e primary outcome was the incidence of RBC
transfusion after the guideline implementation and
the secondary outcomes included the adherence to the
guidelines (Hb threshold of RBC transfusion), ICU and
hospital length of stay (LOS), in-hospital and 28-day
mortality. e sample size was calculated by predicting
the transfusion rate before the implementation using
a guideline about 50% and reduced to 35% during the
post-implementation period, at test signicant level 0.05
and power of 80%. Aer adding up 20% for possible
missing data, the sample size of 200 patients in each
group was required.
Continuous data were presented as median with
interquartile range (IQR) or mean with standard deviation
(SD) with 95% condential interval (CI). Categorical
data were presented as numbers and percentages. Non-
parametric Mann-Whitney U test or unpaired t-test for
continuous data and Chi-Square or Fisher’s exact test for
numbers of events were used for comparison between
pre- and post- educational program. P-value < 0.05 was
considered as statistically signicant. Data analysis was
performed using SPSS 15.0 (SPSS Inc., Chicago, IL).
RESULTS
e overall population was 400 patients, 200 in a
pre-educational group and 200 in a post-educational
group. ree patients from pre-educational group were
excluded due to massive hemorrhage and twelve patients
in post-educational group were excluded due to massive
hemorrhage (10) and incomplete data (2). ese le 197
and 188 patients in pre- and post-educational groups
for analysis. (Fig 2)
Fig 2. e diagram on patient recruitment, inclusion and exclusion
is shown.
Fig 3. e mean Hb and Hct in adherence and non-adherence group
in post-educational patients.
Patient characteristics and intra-operative data were
shown in Table 1. ere was no signicant dierence
in patient characteristics, underlying medical diseases,
Uataya et al.
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TABLE 1. Characteristic of patients on admission to surgical intensive care unit.
Pre-education N=197 Post-education N=188 P-value
Age (year) 64.49 ± 17.86 61.59 ± 19.49 0.13
Gender (male) N (%) 103 (52.3) 105 (55.85) 0.54
Weight (kg) 60.66 ± 15.12 60.28 ± 16.08 0.82
Height (cm) 159.78 ± 8.95 160.55 ± 9.06 0.41
Surgical patients N (%) 180 (91.37) 176 (93.62) 0.44
Medical patients N (%) 17 (8.63) 12 (6.83)
Postoperative day, 0 (0-6) 0 (0-8) 0.53
Median (IQR)
Underlying medical diseases N (%)
Coronary artery disease 31 (15.74) 23 (12.34) 0.4
Vascular disease 34 (17.26) 44 (23.4) 0.16
Respiratory disease 61 (30.96) 62 (32.98) 0.53
End stage renal disease 8 (4.06) 6 (3.19) 0.54
Endocrine disease 57 (28.93) 65 (35.57) 0.27
Stroke 26 (13.2%) 26 (13.8) 0.88
Anemia N (%) 163 (82.74) 146 (77.66) 0.25
Coagulopathy N (%) 40 (20.3) 43 (22.87) 0.62
Immunocompromised N (%) 7 (3.55) 15 (7.98) 0.08
Malignancy N (%) 70 (35.53) 61 (32.45) 0.59
Surgical data N (%)
Elective surgery 124 (62.94) 114 (60.64) 0.28
Emergency surgery 52 (26.4) 63 (33.51)
General surgery 77 (39.1) 73 (38.83) 0.81
Vascular surgery 38 (19.29) 41 (21.8)
Orthopedic surgery 28 (14.21) 22 (11.7)
Obstetric & gynecological surgery 11 (5.58) 13 (6.91)
Type of anesthesia N (%) 135 (68.53) 134 (71.28) 0.05
- General anesthesia 11 (5.58) 3 (1.6)
- Regional anesthesia
ASA class > 2 N (%) 105 (58.33) 119 (63.3) 0.14
Volume of RBC transfusion in 565.5 706.5 0.11
operating room (ml), Median (IQR) (324.8, 924.8) (350.8,1217.8)
APACHE II score 9.67 ± 6.62 8.66 ± 5.5 0.1
SOFA score ,Median (IQR) 2 (0,5) 2 (0,3) 0.07
Indication for SICU admission
Hemodynamic monitoring 196 (99.5) 188 (100) 1.0
Respiratory support 100 (50.76) 120 (63.83) 0.01
Shock
Septic shock 25 (12.7) 18 (9.57) 0.35
Cardiogenic shock 6 (3.05) 2 (1.06)
Hypovolemic shock 11 (5.58) 14 (7.45)
Laboratory on admission
Hb (g/dl) 10.53 ± 2.05 11.57 ± 7.64 0.71
Hct (%) 31.73 ± 6.28 33.24 ± 6.42 0.21
Creatinine (mg/dl) 1.45 ± 1.6 1.32 ± 1.32 0.39
Albumin (g/dl) 2.84 ± 0.69 2.93 ± 2.61 0.66
PaO
2
(mmHg) 177.25 ± 69.06 164.79 ± 79.80 0.2
PaO
2
/FIO
2
ratio 345.05 ± 196.24 359.16 ± 178.33 0.56
Abbreviations: APACHE II = Acute Physiology and Chronic Health Evaluation II score; ASA = American Society of Anesthesiology Physical
Status; CI = condence interval; FiO2 = fraction of inspired oxygen; Hb = hemoglobin; Hct = hematocrit; ;IQR = interquartile range; PaO
2
= partial pressure of arterial oxygenation; RBC = red blood cell; SOFA = Sequential Organ Failure Assessment; SICU = surgical intensive
care unit
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surgical data and patient conditions on ICU admission
between the pre and post- education groups except for
the indication for SICU admission. Respiratory support
as an indication for SICU admission was higher in post-
educational group, than in pre-educational group (63.83%
vs. 50.76%, p = 0.01).
Patients in post-educational group had signicantly
lower RBC transfusion rate (37% vs. 48%, p = 0.02) and
the adherence rate to the guideline was 47% .e mean
Hb level before RBC transfusion was signicantly lower
in post-educational group (8.16 ± 1.43 vs. 8.97 ± 1.57g/
dL, p = 0.001). e 28-day mortality and ICU mortality
were also signicantly lower in post-educational group.
Patients in pre-educational group had signicantly higher
incidence of acute respiratory distress syndrome (ARDS)
(2.5% vs. 0%, p = 0.05) and acute kidney injury (AKI)
(8.1% vs. 2.1%, p = 0.01) than those in post-educational
group (Table 2).
TABLE 2. Transfusion data and outcomes.
Pre-education N=197 Post-education N=188 P-value
Transfusion in SICU, N (%) 96 (48.7) 70 (37.2) 0.02
Threshold
Hb threshold 8.97 ± 1.57 8.16 ± 1.43 0.001
Hct threshold 27.14 ± 4.76 24.54 ± 4.12 < 0.001
RBC volume transfused in SICU, 627.5 650 0.95
Median (IQR) (316.25, 1030.25) (342.5, 904.0)
Outcomes
Ventilator days, Median (IQR) 1 (0,2) 1 (0,2) 0.18
28-day mortality N (%) 22 (11.12) 6 (3.19) 0.007
SICU mortality N (%) 15 (7.61) 3 (1.56) 0.009
SICU LOS, Median (IQR) 2 (1,3) 2 (1,3) 0.73
Hospital LOS, Median (IQR) 15 (9,28) 13 (9,27) 0.37
Complication in SICU,N (%)
Delirium 5 (2.54) 6 (3.2) 0.55
Stroke 0 (0) 4 (2.1) 0.07
Acute myocardial infarction 2 (1.0) 4 (2.1) 0.4
Pulmonary edema 3 (1.52) 2 (1.1) 0.55
ARDS 5 (2.5) 0 (0) 0.05
Acute kidney injury 16 (8.1) 4 (2.1) 0.01
Pneumonia 4 (2.0) 5 (2.7) 0.54
CRBSI 0 (0) 1 (0.5) 0.35
Severe sepsis 6 (3.1) 8 (4.3) 0.48
Septic shock 10 (5.1) 5 (2.7) 0.28
Abbreviations: ARDS = acute respiratory distress syndrome; CRBSI = catheter-related bloodstream infection; Hb = hemoglobin; Hct =
hematocrit; LOS = length of stay; RBC = red blood cell; SICU = surgical intensive care unit; IQR = interquartile range
TABLE 3. Indications for red blood cell transfusion in post-educational group (N = 70).
Indication N (%)
Low Hb 33 (47.1%)
Suspected anemic symptoms 14 (20%)
Keep Hct > 30 % 10 (14.3%)
History of coronary artery disease 8 (11.43%)
MI/Unstable angina 4 (5.7%)
Undened indication 1 (1.4%)
Abbreviations: Hb = hemoglobin; Hct = hematocrit; MI = myocardial infarction
Uataya et al.
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TABLE 4. Laboratory result before red blood cell transfusion in transfused patients of post-educational group
(N = 70).
Laboratory Mean ± SD
Serum lactate (mg/dL) 3.22 ± 2.58
ScvO2 (%) 67 ± 4.24
Base excess (mEq/L) -5.12 ± 5.53
Abbreviation: ScvO
2
= central venous oxygen saturation
DISCUSSION
In this study, the implementation of a RBC transfusion
guideline using the educational program can reduce
the RBC transfusion rate and decrease the hemoglobin
threshold for RBC transfusion. In addition the clinical
outcomes regarding the complication (ARDS, AKI) and
the mortality rate were improved aer the educational
program. Previous study had reported the high incidence
of RBC transfusion up to 83 % in general surgical ICU,
Siriraj hospital.
7
e rate of transfusion is considerably
high in critically ill surgical patients who have been on
mechanical ventilation longer than 48 hours.
7
erefore, the
implementation of a strategy that can reduce the number
of transfusions should be considered. As a result, this study
regarding the implementation of the RBC transfusion
guideline was performed with the aim to decrease the
rate of RBC transfusion. e educational program was
a tool selected to implement the guideline in this study.
Previous study demonstrated that an educational program
coupled with the institution policy for RBC transfusion
could reduce the total RBC units transfused.
17
In this
study, we found that aer the implementation of the
RBC transfusion guideline, the rate of RBC transfusion
was signicantly decreased from nearly 50 % to 37 %
which was comparable to other regions.
15
In addition,
the Hb threshold for RBC transfusion was signicantly
decreased from 9 to 8 g/dL.
Interestingly, the primary and secondary outcomes
were improved, and even the adherence rate to the
transfusion guideline was less than 50%. Several issues
were concerned including 1) physician might accept the
transfusion trigger of Hb around 8-9 g/dL even though
evidences from previous randomized controlled trials
8,11,14
have addressed the safety of restrictive transfusion. A
survey of physician’s attitudes to transfusion practice in
critically ill patients in the United Kingdom
18
reported that
there was signicant variation in transfusion threshold
among physicians. It depended on the scenario and the
severity of patients. e majority of respondents selected a
baseline Hb threshold between 9 and 10 g/dL. In addition,
previous studies from dierent locations reported the
average pre-transfusion Hb at 8.5 g/dL.
15
2) the educational
program regarding the transfusion guideline might not
be frequently provided or reminded. Previous literatures
showed that the implementation of education program
has signicantly decreased RBC transfusion, increased
transfusion safety and helped to prevent the occurrence
of transfusion-related adverse eects.
17,19,20
However,
only the educational program might not be adequate to
improve the outcomes. e combination of the educational
program and the support from medical board in terms of
integration of the guideline recommendation into RBC
transfusion order including the guideline recommendation
can result in the better adherence rate and outcomes
17
3) the plastic surgeon required maintaining the hematocrit
level at least 30 % in the operations which involved ap
procedures, and 5 out of 37 patients (14 %) of non-
adherence were in this category. e researcher did not
argue against this concept, although there were studies
which reported that transfusion trigger of hematocrit
< 25 % can decrease blood transfusion rates without
increasing rates of ap-related complications.
21
; and
4) half of the patients in non-adherence group had
sepsis, and the recent sepsis guideline implemented by
Siriraj hospital recommended the Hb threshold for RBC
transfusion at 10 g/dL.
22
However, this recommendation
was based on the protocol of “early goal-directed therapy
(EGDT)”
23
in sepsis patients that targeted an increase
in mixed venous oxygen saturation to ≥ 70%. is was
achieved by the administration of uid resuscitation, then
RBC transfusion to keep a hematocrit ≥ 30 % and then
administering inotropes (dobutamine). e EGDT group
received more uid and RBC transfusion in the rst 6
hours. e EGDT group demonstrated the improvement
in survival, although it was not possible to separate the
impact of only RBC transfusion strategy on outcome.
Moreover, the recent multicenter RCT
11
, which compared
restrictive and liberal RBC transfusion in patients with
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72
septic shock, did not demonstrate signicant dierence
in terms of mortality between the two groups. Finally,
the recent edition of Sepsis ad septic shock guidelines
was published in 2016. It recommended to reserve RBC
transfusion for patients with a Hb level less than 7 g/dl.
24
Moreover, the updated clinical practice guidelines from
the American Association of Blood Banks (AABB)
recommended that RBC transfusion is not indicated
until the Hb level is 7 g/dL for hospitalized adult patients
who are hemodynamically stable, including critically ill
patients rather than when the Hb is 10 g/dL.
12
Although the adherence rate was not high, the
RBC transfusion rate signicantly decreased aer the
implementation of the guideline. It might not be the
guidelines itself that improved the outcomes, but the
advancement in knowledge regarding the adverse eects of
blood transfusion or the new recommendation from clinical
practice guidelines might be accountable. Nevertheless,
some limitation should be addressed. Twelve patients
(6%) were excluded from the post-educational group
which might alter the adherence rate and outcomes. e
dierent in duration of data collection between pre and
post-educational program should be concerned. e data
of pre-educational group was collected in 2011 whereas the
post-educational information was prospectively collected
during 2014. e dierence in time frame might result
in the variety of treatment modalities especially the RBC
transfusion. e lower incidence of RBC transfusion and
the improvement in mortality rate may result from the
better knowledge from recent publications rather than
the implementation of the guideline. e frequency and
ecacy of educational program as mentioned earlier
might be insucient to change attitude of transfusion.
Other limitation should be addressed including there
was no information regarding detail of transfusion
in pre-educational group especially the indication of
RBC transfusion which might help to interpret the high
transfusion rate. In addition the information regarding
the RBC transfusion in the operating theaters was not
available in both groups, although the hemoglobin at
SICUs admission was not signicantly dierent between
pre and post educational groups.
In conclusion, the implementation of educational
program regarding RBC transfusion guideline can decrease
the RBC transfusion rate.
What is already known on this topic?
Although RBC transfusion is an independent
predictor of death, nosocomial infection, multi-organ
dysfunction syndrome (MODS) and acute respiratory
distress syndrome (ARDS) in critically ill patients, the
majority of the patients admitted to intensive care unit
(ICU) still received RBC transfusion. A pre-transfusion
hemoglobin (Hb) was reported around 8.5 g/dl.
What does this study adds?
The implementation of educational program
regarding RBC transfusion guideline can reduce RBC
transfusion rate, complications and the mortality rates.
e Hb threshold for RBC transfusion was signicantly
decreased from 9 g/dL in the pre-educational group to
8 g/dL in the post-educational group.
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Kesaree Punlee, B.Sc., M.M.*, **, Kamol Udol, M.D., M.Sc.***, *, Vithaya Chaithiraphan, M.D.*, Wandee
Rochanasiri, B.N.S.*, **, Suteera Phrudprisan, B.N.S.*, **, Nithima Ratanasit, M.D.****
*Her Majesty Cardiac Center, **Department of Nursing, Siriraj Hospital, ***Department of Preventive and Social Medicine, ****Department of Medicine,
Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok 10700, ailand.
Prevalence of and Factors Associated with
Inappropriate Indications for Transthoracic
Echocardiography in Adult Outpatients at Siriraj
Hospital
Corresponding author: Kamol Udol
E-mail: kamol.udo@mahidol.ac.th
Received 11 May 2018 Revised 8 October 2018 Accepted 7 November 2018
ORCID ID: 0000-0002-1508-7749
http://dx.doi.org/10.33192/Smj.2019.12
ABSTRACT
Objective: Ordering transthoracic echocardiography (TTE) for inappropriate indications could prolong patient
waiting time, increase echocardiography laboratory workload, and compromise the quality of TTE studies. is
study aimed to investigate the prevalence of and factors associated with inappropriate indications for TTE in adult
outpatients at Siriraj Hospital.
Methods: Two cardiologists prospectively and independently evaluated indications for adult TTE scheduled
during regular oce hours at Her Majesty Cardiac Center, Siriraj Hospital. Cases were classied as appropriate,
inappropriate, or uncertain according to the 2011 Appropriate Use Criteria for Echocardiography proposed by a
group of American cardiovascular organizations. Agreement between the two cardiologists was measured using
weighted kappa statistic, and disagreement was resolved by consensus. Factors were evaluated for association with
inappropriate indications for TTE.
Results: Four hundred and eighty-two patients were included. Weighted kappa statistic was 0.46 [95% condence
interval (CI) 0.34 to 0.59] for agreement between the two cardiologists. Four hundred and thirty-two TTE were
appropriate (89.6%, 95% CI 86.6% to 92.1%), 27 were inappropriate (5.6%, 95% CI 3.9% to 8.0%), and 23 were
uncertain (4.8%, 95% CI 3.2% to 7.0%). Neither status of ordering physician (cardiologist, cardiology fellow, or
cardiothoracic surgeon) nor payment type was found to be signicantly associated with the appropriateness of
indications for TTE.
Conclusion: e prevalence of inappropriate indications among adult outpatients undergoing TTE during regular
oce hours at Siriraj Hospital was low. No signicant association was observed between the appropriateness of
TTE indications and either status of ordering physician or payment type.
Keywords: Transthoracic echocardiography; evaluation of echocardiography; appropriate use criteria; appropriateness
(Siriraj Med J 2019;71: 74-79)
Punlee et al.
INTRODUCTION
Transthoracic echocardiography (TTE) is widely
used in clinical practice due to its safe and non-invasive
nature, its wide availability, and its relatively low cost
compared to other cardiac tests. e demand for TTE
is increasing, and this has resulted in longer waiting
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periods for patients. e number of adult TTE procedures
performed during regular oce hours at Her Majesty
Cardiac Center, Siriraj Hospital, increased from 2,683
per year in 2009 to 3,326 per year in 2012. e average
waiting time for a patient scheduled for outpatient TTE
was 48.9, 74.5, and 58.4 days in 2009, 2010, and 2011,
respectively. is increase in demand for TTE increases
the workload of the echocardiography laboratory, and
this workload increase could compromise the quality of
echocardiographic examinations.
At our echocardiography laboratory, TTE examinations
were previously performed only by cardiologists, or by
cardiology fellows under the supervision of cardiologists.
However, as the number of TTE requests increased over
the years, non-cardiologist cardiac sonographers were
recruited to cope with the increased demand. Even with
the increased number of sonographers, long waiting time
is still a frequent complaint from patients and requesting
physicians. In routine clinical practice, we observed that
some patients were scheduled for TTE with doubtful
or inappropriate indications. Reducing the number of
TTE procedures that are unnecessarily performed due to
inappropriate indications may help to shorten the waiting
period for TTE, reduce echocardiography laboratory
workload, improve the quality of TTE examinations, and
reduce healthcare costs. Experts in echocardiography have
proposed clinical situations in which echocardiography
should be considered indicated as a guideline for the
appropriate and eective use of echocardiography in
clinical practice.
1,2
Reports from Western countries showed
that 56% to 92% of echocardiographic examinations
were considered appropriate,
3-10
and that non-cardiac
specialists were more likely to inappropriately request
echocardiography.
4,6,8
In ailand, a study published
in 2011 from a university hospital found that 89% of
echocardiographic requests among inpatients and
outpatients were appropriate
11
when evaluated against
the 2007 Appropriateness Criteria for Transthoracic and
Transesophageal echocardiography.
1
e aim of this
study was to investigate the prevalence of and factors
associated with inappropriate indications for TTE in
adult outpatients at Siriraj Hospital according to the
updated Appropriate Use Criteria for Echocardiography
published in 2011.
2
MATERIALS AND METHODS
Methods
We conducted this prospective study to assess the
appropriateness of TTE requests among adult outpatients
scheduled for TTE at the echocardiography laboratory of
Her Majesty Cardiac Center, Faculty of Medicine Siriraj
Hospital, Mahidol University, Bangkok, ailand. e
Siriraj Institutional Review Board (SIRB) approved the
protocol for this study (Si 067/2013), and all patients
provided written informed consent before enrollment
into the study.
Study population
e principal inclusion criterion was adult outpatients
electively scheduled for TTE during regular office
hours. Patients were prospectively and consecutively
enrolled. Patients scheduled for stress echocardiography,
transesophageal echocardiography, echocardiography
performed for other research purposes, and TTE performed
aer oce hours were excluded.
Study procedures
Two cardiologists (KU and VC) independently
reviewed the medical record of each patient to determine
the indication for TTE. Indications were classied as
appropriate, inappropriate, or uncertain according to
the 2011 Appropriate Use Criteria for Echocardiography
proposed by the American College of Cardiology Foundation
(ACCF) in collaboration with various other American
cardiovascular organizations.
2
ere are 98 criteria listed
in the appropriate use criteria for TTE, 57 of which are
considered appropriate, 29 inappropriate, and 12 uncertain.
If the indication for TTE in a particular patient did not
match any of the 98 criteria, each of the two reviewing
cardiologists used his own judgment to determine the
appropriateness of the indication. Disagreement between
the two cardiologists was resolved by discussion and
consensus. Data relating to the status of the physician
that ordered the TTE, and the type of payment to cover
the cost of the TTE were also recorded. Regarding
physician status, only cardiology fellows, cardiologists,
and cardiothoracic surgeons are authorized to order TTE
at our center. Payment type was categorized into any
one of 3 ailand health insurance schemes or self-pay.
Statistical Analysis
e primary objective of this study was to estimate the
proportion of TTE that were ordered with inappropriate
indications among adult outpatients electively scheduled
for TTE during regular oce hours. We estimated that
a minimum of 480 subjects would be required to ensure
that the limits of the 2-sided 95% condence interval
(CI) of this proportion would not exceed ±0.045 from
the observed proportion. We chose to use an expected
proportion of inappropriate TTE of 0.5 in the sample size
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calculation, as this value would yield the largest variance
of the estimate of proportion, thereby maximizing the
required sample size.
Demographic data, status of ordering physician,
and payment type were summarized using descriptive
statistics. Continuous variables are presented as median
and interquartile range (IQR), and Kruskal-Wallis test
was used to compare continuous variables between
groups. Categorical variables are reported as number and
percentage, and Fisher’s exact test was used to compare
categorical variables between groups. Agreement between
the two cardiologists was measured using weighted
kappa statistic with linear weights. All statistical tests
were carried out at the 2-sided signicance level of 0.05.
RESULTS
A total of 482 patients were consecutively enrolled in
this study between March 1 and June 11, 2013. Demographic
data, status of ordering physician, and payment type of
study participants are shown in Table 1. Almost half
(47.1%) of subjects were male. Patient age ranged from
16 to 95 years. Cardiologists or cardiology fellows ordered
TTE in about 95% of cases. e Civil Servant Medical
Benets system and the National Health Insurance system
covered the cost of TTE in 40.7% and 33.8% of patients,
respectively.
e two cardiologists concordantly determined
the indications for TTE as appropriate in 409 subjects,
as inappropriate in 12 subjects, and as uncertain in 11
subjects, representing the observed agreement of 89.6%.
When chance agreement was taken into account, there
was moderate agreement between the two cardiologists
regarding the level of appropriateness of TTE indications
(weighted kappa 0.46, 95% CI 0.34 to 0.59). Consensus
was achieved in all 50 cases where there was initial
disagreement between reviewers.
Aer consensus was reached, the indications for TTE
were considered appropriate in 432 patients (89.6%, 95%
CI 86.6% to 92.1%), inappropriate in 27 patients (5.6%,
95% CI 3.9% to 8.0%), and uncertain in 23 patients (4.8%,
95% CI 3.2% to 7.0%). ere were six patients whose
indications for TTE were not specically listed in the
2011 Appropriate Use Criteria for Echocardiography, so
determination of the level of appropriateness was based
on judgement of the reviewing cardiologists. Among those
6 indications, 1 was judged appropriate, 2 inappropriate,
and 3 uncertain.
ere was a statistically signicant dierence in age
among the appropriate, inappropriate, and uncertain TTE
indication groups. Subjects in the uncertain group were
signicantly younger than those in the appropriate and
inappropriate groups (p = 0.003; Table 2). Females were
signicantly more likely than males to have TTE with
appropriate indications (92.9% vs. 85.9%, p = 0.043).
Although cardiology fellows had a higher proportion
of appropriate indications for TTE request (92.7%)
than cardiologists (86.0%) and cardiothoracic surgeons
(85.7%), there was no statistically signicant association
between the status of the ordering physician and the level
of TTE indication appropriateness (p = 0.062). Similarly,
patient payment type was not found to be signicantly
associated with the level of indication appropriateness
(p = 0.071).
TABLE 1. Characteristics of study participants.
Variables Value (n = 482)
Age (years), median (IQR) 61 (50, 71)
Male, n (%) 227 (47.1%)
Ordering physician, n (%)
Cardiologist 200 (41.5%)
Cardiology fellow 261 (54.1%)
Cardiothoracic surgeon 21 (4.4%)
Payment type, n (%)
CivilServantMedicalBenets 196(40.7%)
National Health Insurance 163 (33.8%)
Social Security 48 (10.0%)
Self-pay 75 (15.5%)
Abbreviation: IQR = interquartile range
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TABLE 2. Comparison of various factors among dierent levels of appropriateness relative to indications for adult
outpatient transthoracic echocardiography during regular oce hours at Siriraj Hospital.
Variables
a
Appropriate Inappropriate Uncertain P-value
b
Age (years), median (IQR) 62 (50, 72) 61 (54, 72) 43 (33, 63) 0.003
c
Gender, n (%) 0.043
Male 195(85.9%) 17(7.5%) 15(6.6%)
Female 237(92.9%) 10(3.9%) 8(3.1%)
Ordering physician, n (%) 0.062
Cardiologist 172 (86.0%) 17 (8.5%) 11 (5.5%)
Cardiologyfellow 242(92.7%) 8(3.1%) 11(4.2%)
Cardiothoracicsurgeon 18(85.7%) 2(9.5%) 1(4.8%)
Payment type, n (%) 0.071
CivilServantMedicalBenets 176(89.8%) 14(7.1%) 6(3.1%)
NationalHealthInsurance 150(92.0%) 6(3.7%) 7(4.3%)
SocialSecurity 38(79.2%) 3(6.3%) 7(14.6%)
Self-pay 68(90.7%) 4(5.3%) 3(4.0%)
a
Percentage data are shown as row percentage.
b
Fisher’s exact test, except where indicated otherwise
c
Kruskal-Wallis test
Abbreviation: IQR = Interquartile range
e most frequent indication for TTE, according to
the 2011 Appropriate Use Criteria for Echocardiography,
was presentation with symptoms or conditions potentially
related to suspected cardiac etiology, such as chest pain,
shortness of breath, and palpitations (94 patients, 19.5%;
Table 3), followed by initial evaluation for suspected
valvular or structural heart disease (48 patients, 10.0%;
Table 3). Among the 27 patients with inappropriate
indications, the most frequently observed indication was
routine surveillance of ventricular function in clinically
stable subjects with known coronary artery disease (5
patients, 18.5%; Table 4).
TABLE 3. e 10 most common indications for adult outpatient transthoracic echocardiography during regular
oce hours at Siriraj Hospital.
Indication n (%)
(Total n = 482)
1. Symptomsorconditionspotentiallyrelatedtosuspectedcardiacetiologyincludingbutnot 94(19.5%)
limited to chest pain, shortness of breath, palpitations, transient ischemic attack, stroke,
or peripheral embolic event
2. Initial evaluation when there is a reasonable suspicion of valvular or structural heart disease 48 (10.0%)
3. Re-evaluation of known valvular heart disease with a change in clinical status or cardiac exam 36 (7.5%)
or to guide therapy
4. Prior testing that is concerning for heart disease or structural abnormality including but not 35 (7.3%)
limited to chest X-ray, baseline scout images for stress echocardiogram, ECG, or cardiac biomarkers
5. Routinesurveillance(≥1year)ofmoderateorseverevalvularregurgitationwithoutchange 31(6.4%)
in clinical status or cardiac exam
6. Sustainedornonsustainedatrialbrillation,supraventriculartachycardia,orventriculartachycardia 30(6.2%)
7. Initialevaluationofknownorsuspectedheartfailure(systolicordiastolic)basedonsymptoms, 29(6.0%)
signs, or abnormal test results
8. Initial evaluation of ventricular function following acute coronary syndrome 26 (5.4%)
9. Routinesurveillance(≥1year)ofmoderateorseverevalvularstenosiswithoutachangein 18(3.7%)
clinical status or cardiac exam
10. Evaluation of suspected pulmonary hypertension including evaluation of right ventricular function 15 (3.2%)
and estimated pulmonary artery pressure
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TABLE 4. Common inappropriate indications for adult outpatient transthoracic echocardiography during regular
oce hours at Siriraj Hospital.
Indication n (%)
(Total n = 27)
1. Routine surveillance of ventricular function with known coronary artery disease and 5 (18.5%)
no change in clinical status or cardiac exam
2. Initial evaluation of ventricular function (e.g., screening) with no symptoms or signs of 2 (7.4%)
cardiovascular disease
3. Routine perioperative evaluation of ventricular function with no symptoms or signs of 2 (7.4%)
cardiovascular disease
4. Routine surveillance (< 1 year) of moderate or severe valvular stenosis without a change 2 (7.4%)
in clinical status or cardiac exam
5. Routine surveillance (< 3 year after valve implantation) of prosthetic valve if no known 2 (7.4%)
or suspected valve dysfunction
6. Routine evaluation of systemic hypertension without symptoms or signs of hypertensive 2 (7.4%)
heart disease
DISCUSSION
We evaluated the indications for TTE among adult
outpatients undergoing TTE during regular oce hours,
and determined their appropriateness according to the
2011 Appropriate Use Criteria for Echocardiography.
Our findings revealed that most of outpatient TTE
examinations performed at our center during the study
period were appropriate, regardless of the status of the
ordering physician or the payment type used to cover
the cost of TTE.
Our result is very comparable to that reported from
another university hospital in ailand (89% appropriate TTE
indications) that enrolled both inpatients and outpatients,
and that based the appropriateness of TTE indications
on 2007 Appropriateness Criteria for Transthoracic and
Transesophageal Echocardiography.
11
In that study, the
level of appropriateness of TTE requests was similar
between inpatients and outpatients. e most common
inappropriate indications for TTE were preoperative
evaluation (50% of inappropriate TTE), as determined
by expert opinion since this indication is not listed as
an indication in the 2007 Appropriateness Criteria for
Transthoracic and Transesophageal Echocardiography, and
evaluation of endocarditis without evidence of bacteremia
or new murmur (19% of inappropriate TTE). In our
study, the most common inappropriate indications
for TTE were evaluation of le ventricular function in
clinically stable coronary artery disease patients, and
evaluation of asymptomatic individuals without suggestive
evidence of cardiovascular disease (26% of inappropriate
TTE for both indications combined). TTE performed
for perioperative evaluation was found in only 7% of
inappropriate TTE in our study, probably because this
indication is clearly listed as inappropriate in the 2011
Appropriate Use Criteria for Echocardiography. We
did not have any TTE evaluation for endocarditis in our
study, because we included only elective outpatients.
e rate of appropriate TTE requests at our hospital,
and probably at most university hospitals in ailand, is
higher than the rates reported from the United States of
America (56% to 92%).
3-10
is may be explained in part by
the policy imposed at our center, and at other university
hospitals in ailand, to limit the privilege of ordering
TTE to only cardiovascular specialists. At many centers
in the US, family physicians and general practitioners
can order TTE, and the rates of inappropriate requests
made by these physicians are usually higher than those
made by cardiovascular specialists.
4,6,8
Another possible
explanation for the observed high rate of appropriate TTE
indications is that ordering physicians may occasionally
have to perform echocardiographic examination themselves
without being paid extra to do so. is may have the
eect of inuencing requests for TTE that are more
accurately based on guideline recommendations.
e observed tendency of higher rate of appropriate
requests among cardiology fellows compared to cardiologists
and cardiothoracic surgeons might reect the nature
of practice during training at an academic institution.
e management decisions of cardiology fellows are
usually monitored by certied cardiologists, and this
may inspire them to be more cautious and more likely
to consult their mentor before ordering a test.
is study was inspired by the idea that decreasing
or eliminating inappropriate TTE requests would lead to
Punlee et al.
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improvement in patient waiting time for, and perhaps the
quality of TTE. However, the magnitude of inappropriate
requests found in this study was only 5.6%, which indicates
that the practice at our center regarding the use of TTE
is reasonable, and that only modest improvement would
be possible. As such, alternative strategies need to be
explored in order to improve outpatient TTE service.
Some limitations of this study need to be mentioned.
First, included patients were enrolled over a relatively
short 14-week study period, so the indications that were
identied may not reect all of the TTE indications that
are used in clinical practice. Second, even though our
reviewers are both board-certied cardiologists, which
gave our study more credibility than if we had enlisted
non-cardiologist reviewers, their agreement was only
moderate, which suggests the complexity associated with
reviewing medical records to identify TTE indications. e
dierences between reviewers are likely due to incomplete
documentation on echocardiography request forms
and/or medical records. ird and last, the results of
this study may not be generalizable to other centers
with dierent TTE request system practices, such as
non-teaching hospitals or private hospitals.
CONCLUSION
e prevalence of inappropriate indications among
adult outpatients undergoing TTE during regular oce hours
at Siriraj Hospital was low. No signicant association was
observed between the appropriateness of TTE indications
and either status of ordering physician or payment type.
ACKNOWLEDGMENTS
is study was funded by a grant from the Siriraj
Research Development Fund (managed by the Routine to
Research Project), Faculty of Medicine Siriraj Hospital,
Mahidol University, Bangkok, ailand.
Potential conict of interest: All authors declare no
personal or professional conicts of interest, and no
nancial support from companies that produce and/
or distribute the drugs, devices, or materials described
in this report.
REFERENCES
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2. Douglas PS, Garcia MJ, Haines DE, Lai WW, Manning WJ,
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3. Barbier P, Alimento M, Berna G. Clinical utility of guideline-based
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4. Ward RP, Mansour IN, Lemieux N, Gera N, Mehta R, Lang RM.
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echocardiography. JACC Cardiovasc Imaging. 2008;1:663-71.
5. Kirkpatrick JN, Ky B, Rahmouni HW, Chirinos JA, Farmer SA,
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2009;22:53-9.
6. Willens HJ, Gómez-Marín O, Heldman A, Chakko S, Postel C,
Hasan T, et al. Adherence to appropriateness criteria for
transthoracic echocardiography: comparisons between a regional
department of Veterans Aairs health care system and academic
practice and between physicians and mid-level providers. J
Am Soc Echocardiogr. 2009;22:793-9.
7. Parikh PB, Asheld J, Kort S. Does the revised appropriate use
criteria for echocardiography represent an improvement
over the initial criteria? A comparison between the 2011 and
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Am Soc Echocardiogr. 2012;25:228-33.
8. Ballo P, Bandini F, Capecchi I, Chiodi L, Ferro G, Fortini A,
et al. Application of 2011 American College of Cardiology
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9. Bailey SA, Mosteanu I, Tietjen PA, Petrini JR, Alexander J, Keller
AM. e use of transthoracic echocardiography and adherence to
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Echocardiogr. 2012;25:1015-22.
10. Patil HR, Coggins TR, Kusnetzky LL, Main ML. Evaluation
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11. Satitthummanid S, Songmuang SB, Suithichaiyakul T. Evaluation
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Mongkol Laohapensang, M.D.*, Tipsuda Tangsriwong, M.D.**, Niramol Tantemsapya, M.D.*
*Department of Surgery, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, 10700, **Department of Surgery, Buddhachinaraj
Phitsanulok Hospital, Phitsanulok 65000, ailand
Esophageal Replacement in Children: A 10-Year,
Single-Center Experience
Corresponding author: Mongkol Laohapensang
E-mail: mongkol.lao@mahidol.ac.th
Received 26 October 2018 Revised 31 January 2019 Accepted 1 February 2019
ORCID ID: 0000-0002-0774-5705
http://dx.doi.org/10.33192/Smj.2019.13
ABSTRACT
Objective: Various esophageal replacement gras have been used in children, although none can equal the native
esophagus. e purpose of this study was to review the complications and outcomes associated with using dierent
techniques in a single institute.
Methods: A retrospective medical record review was conducted from 2006 to 2016. Patient demographics, perioperative
clinical courses, complications and long-term outcomes were reported as percentages and categorized according
to the surgical procedure performed.
Results: A total of 15 children underwent esophageal replacement procedures, comprising 7 (47%) isoperistaltic
gastric tubes, 3 (20%) colonic interpositions, 3 (20%) gastric transpositions and 2 (13%) reversed gastric tubes.
Indications for esophageal replacement included long-gap esophageal atresia (5; 33%), esophageal atresia with
severe postoperative complications (6; 40%), and caustic injury (4; 27%).e mean age of patients was 2.9 years
(range: 0.2–15 years). e average follow-up duration was 3.6 years (range: 0.4–8 years). ere was no perioperative
mortality and no gra loss in any group. e long-term outcomes were acceptable, with no late stricture. Eighty-
six percent of the patients in the isoperistaltic gastric tube group and all patients in the other procedural groups
achieved full oral feeding. Nevertheless, the patients had various degrees of malnutrition.
Conclusion: Esophageal replacement remains a major challenge in children. Our experience indicates that children
can be safely operated on using any of these methods, with acceptable outcomes and no deaths. Nevertheless, the
long-term consequences and complications should be monitored throughout adulthood.
Keywords: Esophageal replacement; isoperistaltic gastric tube; reversed gastric tube; gastric transposition; colonic
interposition (Siriraj Med J 2019;71: 80-88)
Laohapensang et al.
INTRODUCTION
Over the last several decades, many dierent techniques
and various visceral substitute gras have been used
for esophageal replacement in children.
1-4
Common
indications for the esophageal replacement procedure
in children include long-gap esophageal atresia, severe
peptic or caustic injuries, and anastomotic strictures.
5-7
e ideal esophageal substitute should closely imitate the
native esophagus both in size and function; nevertheless,
none can match a normal esophagus.
1,5
Several studies
have reported comparable outcomes for each technique,
with no signicant dierences in terms of their early and
late complications.
1-7
e procedure selection and gra
choice in those studies were based on the anatomy and
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availability of the visceral substitute and, in particular,
the experience and preferences of the surgeon. Factors
inuencing the outcomes were related to the infrequency
with which the procedures were performed, the variable
expertise among the surgeons, and the lack of an ideal
conduit.
5
e aim of the present study was to review our
10-year-experience using various esophageal replacement
techniques and to compare the complications and outcomes
of those procedures in children at the Division of Pediatric
Surgery, Faculty of Medicine Siriraj Hospital, Mahidol
University, Bangkok, ailand.
Ethical approval for this study as a retrospective
evaluation of practice was obtained from the Siriraj
Institutional Review Board (Si 628/2016).
MATERIALS AND METHODS
e medical records of all patients who underwent an
esophageal replacement procedure between 2006 and 2016
were retrospectively reviewed. e clinical data extracted
included patient demographics, indications for surgery,
perioperative courses, complications and outcomes. Each
parameter was reported as a percentage and categorized
according to the surgical procedure performed. e
patient demographics included the associated congenital
anomalies and pre-replacement surgical procedures. e
early postoperative outcomes were evaluated according
to the duration of the overall admissions, the degree of
intensive care and intubation needed, and the number
of readmissions aer the replacement operation. e
perioperative adverse events comprised respiratory
complications, esophageal leakages, esophagocutaneous
stulas, gut obstructions, delayed gastric function, dumping
syndrome, wound infections and death; they were reported
as percentages for each surgical procedure. e operative
outcomes were categorized into gastrointestinal and
respiratory outcomes and listed in the same fashion. e
long-term anthropometric assessments were expressed
in degrees of malnutrition, using the weight at the nal
follow-up compared to the standard growth chart.
RESULTS
Between January 2006 and December 2016, 15
patients underwent an esophageal replacement at the
hospital. e patients had mostly been referred from
other hospitals. e mean age of the patients was 2.9 years
(range: 0.2-15 years). ere were 11 (73%) males and 4
(27%) females. e average follow-up duration was 3.6
years (range: 0.4-8 years). e operative procedures of
the 15 children studied consisted of 7 (47%) isoperistaltic
gastric tubes, 3 (20%) colonic interpositions, 3 (20%)
gastric transpositions and 2 (13%) reversed gastric tubes.
e indications for esophageal replacement included 5
(33%) long-gap esophageal atresias, 6 (40%) esophageal
atresias with postoperative complications, and 4 (27%)
caustic injuries. e indications for surgery, categorized
by procedure, are demonstrated in Table 1.
e majority of patients (73%) had esophageal
atresia, either with long-gap or severe postoperative
complications aer esophagoesophagostomy (anastomotic
leakages, strictures unresponsive to dilatation, empyema
thoracis and diverticulum with bleeding), whereas 4
patients (27%) had caustic injuries.
e patient characteristics and associated anomalies
are summarized in Table 2. e median age at surgery
was 12 months for those patients who underwent the
gastric tube and gastric transposition procedures, but
much older (108 months) for the colonic interposition
group. Accordingly, the median body weights at surgery
were 9.6 and 8 kg for the isoperistaltic gastric tube and
gastric transposition groups, respectively. e smallest
baby in each group weighed 4.7 and 6.2 kg, respectively.
ere were only 2 cases in the reversed gastric tube
group weighing 12 and 21 kg at the time of surgery. As
the colonic interposition procedure was performed in
older children, their median weight was 20 kg. VACTERL
association (vertebral, anorectal, cardiac, renal and limbs
anomalies) was the most common associated anomaly,
due to esophageal atresia (types A, B and C) being the
main indication for surgery.
TABLE 1. Indications for esophageal replacement.
Diagnosis Isoperistaltic gastric Reversed gastric Gastric Colonic
tube tube transposition interposition
Long-gap EA 4 - - 1
EA with complications 2 1 3 -
Caustic injury 1 1 - 2
Total 7 (47%) 2 (13%) 3 (20%) 3 (20%)
Abbreviation: EA= esophageal atresia
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TABLE 2. Patient characteristics.
Isoperistaltic Reversed Gastric Colonic
gastric tube gastric tube transposition interposition
(n = 7) (n = 2) (n = 3) (n = 3)
Median age at surgery
(months) 12 (3–120) 12, 120 12 (6–24) 108 (36–180)
BW at surgery (kg) 9.6 (4.7–27) 12, 21 8 (6.2–10) 20 (17–23)
Caustic injury 1 (14%) 1 (50%) - 2 (67%)
Type atresia 6 (86%) 1 (50%) 3 (100%) 1 (33%)
Nostula(typeA) 2(33%) - - 1(100%)
Proximalstula(typeB) 2(33%) - - -
Distalstula(typeC) 2(33%) 1(100%) 3(100%) -
Congenital anomaly
Vertebral - - 2 (67%) 1 (33%)
Cardiac 2 (28%) - 3 (100%) -
Anorectal - - 1 (33%) -
Renal - - 1 (33%) -
Limbs - - 1 (33%) -
Duodenal atresia 1 (14%) - 1 (33%) -
Down syndrome 1 (14%) - - -
Pre-replacement surgery
Esophagoesophagostomy 2 (28%) 1 (50%) 2 (67%) -
Gastrostomy 7 (100%) 2 (100%) 3 (100%) 2 (67%)
Cervical esophagostomy 6 (86%) 1 (50%) 3 (100%) -
Endoscope with dilatation 2 (28%) 1 (50%) - 2 (67%)
Gastrojejunostomy - - - 1 (33%)
Duodenoduodenostomy 1 (1–4%) - - -
All patients with long-gap esophageal atresia
unsuitable for elongation techniques underwent cervical
esophagostomy and feeding gastrostomy prior to their
replacement surgery. Other types of esophageal atresia
patients initially underwent esophagoesophagostomy
and subsequently required cervical esophagostomy and
gastrostomy due to severe complications, as mentioned
above. In the case of children with caustic strictures, 4
(27%) underwent esophageal replacement when their
endoscopic dilatations failed. Almost all patients (93%)
were fed by gastrostomy until the appropriate time and
weight for the replacement procedure.
e early postoperative outcomes are presented in
Table 3. e gastric transposition group had the longest
median admission period of 68 days owing to previous
operations and complications. e intensive care periods
after the replacement operation for all groups were
typically 2 to 3 weeks. e median intubation period for
these techniques was about 2 to 6 days. Postoperative
readmissions occurred approximately 3 to 5 times for
esophagoscopy and anastomotic dilatation, although
some were due to respiratory problems.
e perioperative complications are summarized in
Table 4. ere was no perioperative mortality, and none
of the gras were lost. Respiratory complications were the
most common adverse event in all groups. ere was 1
esophageal leakage (in the gastric transposition group) and
2 esophageal stulas (one each in the gastric transposition
and isoperistaltic gastric tube groups). e complications
of delayed gastric function and dumping syndrome arose
aer gastric transposition, with an incidence of 33%
each. Wound infections developed most frequently in
the colonic interposition group, aecting all of its cases.
All esophageal leakage, esophagocutaneous stulas and
gut obstructions were successfully treated conservatively.
e gastrointestinal and respiratory outcomes are listed
in Table 5. Full oral feeding was achieved by 86% of the
isoperistaltic gastric tube group and 100% of the other
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TABLE 3. Early postoperative outcomes.
Isoperistaltic Reversed Gastric Colonic
gastric tube gastric tube transposition interposition
(n = 7) (n = 2) (n = 3) (n = 3)
Admission period 37 (17–106) 18, 27 68 (24–84) 22 (20–55)
Intensive care period 17 (8–44) 8, 15 23 (9–35) 13 (4–14)
Intubation period 2 (1–9) 1, 2 6 (1–10) 2 (1–12)
Postoperative readmissions 5 (1–14) 2, 12 5 (2–20) 3 (1–3)
(number of readmissions)
Values expressed as days: median (range).
TABLE 4. Perioperative complications.
Isoperistaltic Reversed Gastric Colonic
gastric tube gastric tube transposition interposition
(n = 7) (n = 2) (n = 3) (n = 3)
Respiratory complications 1 (14%) 2 (100%) 3 (100%) 1 (33%)
Pneumothorax - 1 (50%) - -
Pneumonia or aspiration 1 (14%) 1 (50%) 3 (100%) 1 (33%)
Esophageal leakage - - 1 (33%) -
Esophagocutaneousstula 1(14%) - 1(33%) -
Gut obstruction 2 (28%) - - 1 (33%)
Delayed gastric function - - 1 (33%) -
Dumping syndrome - - 1 (33%) -
Wound infection 1 (14%) - - 3 (100%)
Death - (0%) - (0%) - (0%) - (0%)
TABLE 5. Gastrointestinal and respiratory outcomes of esophageal replacement.
Isoperistaltic Reversed Gastric Colonic
gastric tube gastric tube transposition interposition
(n = 7) (n = 2) (n = 3) (n = 3)
Gastrointestinal
Full oral feeding 6 (86%) 2 (100%) 3 (100%) 3 (100%)
Anastomotic stenosis 6 (86%) 1 (50%) 3 (100%) 3 (100%)
Number of endoscopic
dilatations per patient 2 (2–11) 6 4 (2–10) 2 (1–31)
Dysphagia 4 (57%) 1 (50%) 1 (33%) 2 (67%)
Oromotor dysfunction - 1 (50%) 3 (100%) -
Reux 1(14%) - 1(33%) 1(33%)
Tortuous/redundant 2 (28%)* 1 (50%)* - 3 (100%)
Respiratory
Chronic lung disease 1 (14%) - - -
Recurrent pneumonia 1 (14%) 1 (50%) 3 (100%) -
Restrictive lung disease - - - 1 (33%)
*e tortuosity of the gras was surgically corrected by manubrium excision
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groups. Anastomotic stenosis was found in 50%-100% of
the cases in the groups, and all resolved aer endoscopic
dilatation. e median number of endoscopic dilatations
required ranged from 2-6 times per patient. Dysphagia
and reux manifested sporadically in 14%-67% of the
cases in the groups. Oromotor dysfunction developed
in all patients who underwent gastric transposition.
Nevertheless, the majority of patients achieved full oral
feeding. Tortuosity of the cervical anastomosis arose in
28%-50% of cases, and mainly in the isoperistaltic and
reversed gastric tube groups; all cases were surgically
corrected by manubrium resection. Redundancy of the
gra was present in the colonic interposition group
with minimal symptoms; therefore, no intervention was
required. e respiratory outcomes comprised chronic
lungs, restrictive lung disease and, the most common,
recurrent pneumonia (which was found in all patients
in the gastric transposition group).
e long-term anthropometric outcomes at the
nal follow-up are illustrated in Table 6 and Fig 1. e
patients had various degrees of malnutrition, particularly
in the gastric tube and gastric transposition groups, with
57%-100% of the aforementioned groups weighing less
than the third centile on the standard growth chart. An
average weight gain was achieved within the study period
by 67% of the patients in the colonic interposition group
and 20% of the study cohort.
DISCUSSION
Esophageal replacement remains a major challenge
in children. Since there is presently no replacement
technique that can replace the features of a normal
esophagus, many pediatric surgeons believe patients
are best served by their own esophagus. e common
indications in children are long-gap esophageal atresia;
severe peptic ulcers; caustic and anastomotic strictures; and
some rare esophageal disorders such as tumor, prolonged
impaction of radiolucent foreign bodies and intractable
achalasia.
5,6,8,9
e common indications for replacement
procedure in our study were long-gap esophageal atresia
(types A and B) and complicated esophageal atresia with
distal tracheoesophageal stula. Caustic injuries following
failed dilatations are another common indication for
esophageal replacement in children. As in other developing
countries
10
, caustic injuries in ailand continue to
be one of the most common health hazards because
caustic agents, such as household cleaning materials,
are frequently stored in unsuitable or poorly labelled
secondary containers.
Various esophageal replacement gras and techniques
have comparable outcomes, none of which emulate the
normal esophagus.
11
e ideal esophageal conduit should
maintain the entire esophageal length, which would
allow normal swallowing while technically being simple
and adaptable for small children. Accordingly, it should
not compress the mediastinum or suppress respiration,
should not become tortuous or redundant, should have
minimal gastro-esophageal reux, should not increase
the malignancy risk, and should function normally for
the lifetime of the patient.
5,10
Many esophageal replacement techniques are being
practiced and recommended without clear consensus.
Furthermore, no randomized, controlled trials have yet
demonstrated signicant, superior, clinical outcomes of
any one of the dierent types of esophageal replacement.
e four most commonly used esophageal replacement
techniques are gastric transposition, gastric tube interposition
(isoperistaltic or reversed), colonic interposition and
TABLE 6. Long term anthropometric outcomes at nal follow up.
Isoperistaltic Reversed Gastric Colonic
gastric tube gastric tube transposition interposition
(n = 7) (n = 2) (n = 3) (n = 3)
Malnutrition 4 (57%) 2 (100%) 2 (67%) 1 (33%)
Percentile body weight
< 3 percentile 4 (57%) 2 (100%) 2 (67%) 1 (33%)
3-25 percentile 2 (28%) - 1 (33%) -
25-75 percentile 1 (14%) - - 2 (67%)
75-97 percentile - - - -
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Fig 1. Growth at nal follow-up.
jejunal interposition.
8,11–15
At our institute, the more
technically demanding jejunal interposition has been the
least preferred; therefore, the esophageal replacement
procedures practiced have comprised gastric transposition,
isoperistaltic or reversed gastric tube, and colonic
interposition. Almost half (47%) of the patients studied
at our institute underwent isoperistaltic gastric tube,
given that the stomach has better vascularity and fewer
ischemic complications.
1,11
e gra choices were based
on anatomical considerations and the preferred technique
of the treating surgeons rather than on any discernible
objective data. e factors inuencing the outcomes were
related to the relatively infrequent need for esophageal
replacement, the surgeons’ expertise and the absence
of ideal gras.
5
Various esophageal substitutes have different
advantages, technical diculties and specic complications.
e overall morbidity is high, the most common being
anastomotic leakage and stricture, with both ranging
from 10% to 20%.
8
e overall replacement-related mortality rate has
been assessed to be approximately 2%.
5
In our study,
there was no perioperative mortality, and none of the
gras were lost. e early post-operative outcomes of the
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86
methods showed no dierences in their intubation periods,
intensive care periods or post-operative readmission
rates. e intubation period was about 1-2 days, whereas
the intensive care period was typically about two weeks.
Patients in the gastric transposition group had the longest
median intubation and intensive care periods (6 and 23
days, respectively), being complicated by type-C esophageal
atresia from previous esophagoesophagostomy operations
with associated cardiac anomalies. e post-operative
readmissions of all groups were due to pneumonia or
esophageal dilatation, and they averaged around 3-5
times per group.
Gastric tube interposition gras are constructed from
the greater curvature in a reversed (antiperistaltic) or
isoperistaltic fashion.
9
Tube gra necrosis is rare because
the gastric tube has an excellent and reliable blood supply
from the submucosal plexus and gastroepiploic vessels.
1
e gastric tube can bridge relatively long gaps and remain
as a passive conduit with a tubular shape and without
dilatation or tortuosity. Other series using a gastric
tube substitute had cervical anastomotic leakage despite
being sealed spontaneously with dilatable strictures.
1,16
In our study, the most common perioperative adverse
event was respiratory complications, appearing in both
isoperistaltic and reversed gastric tube interpositions.
Esophagocutaneous stulas (14%), gut obstructions
(28%) and wound infections (14%) were not common
aer isoperistaltic gastric tube interposition, and all were
resolved by conservative treatment.
e evaluation of the long-term gastrointestinal
outcomes found that 86% of the patients in the isoperistaltic
gastric tube interposition group and all patients in the
other groups achieved full oral feeding. Anastomotic
stenosis was common, albeit dilatable in all groups.
Borgnon et al.
1
reported that of their series of esophageal
replacements with isoperistaltic gastric tube, 80% achieved
a normal diet, 15% had mild dysphagia, 15% had major
dysphagia, and 10% had redundant gras with dumping
syndrome and cervical Barrett’s esophagus. Our study
demonstrated that 57% of the patients had dysphagia,
14% had reux and 28% had tortuosity of the cervical
anastomosis. In consideration of the proximal esophageal
gra anastomosis, most could be achieved through the
neck incision. When the anastomosis is high within
the neck, the thoracic inlet can act as a constriction to
the anastomosis. In that event, the thoracic inlet can be
widened by resecting the upper part of the manubrium
or the sternal head of the le clavicle. Consequently,
the tortuous gras in 3 patients who underwent gastric
tube interposition procedures were surgically corrected
by manubrium excision. Although the incidences of
long-term respiratory problems, including recurrent
pneumonia and chronic lung diseases, ranged from
14%-50%, they did not dier between the two gastric
tube groups.
e disadvantage of the gastric tube is an extensive
suture line that produces a higher incidence of leakages
and strictures. e reduced gastric capacity and the
production of acid within the tube gra results in acid
reux into the cervical esophagus, precipitating Barrett’s
esophagitis.
1,5,16,17
Esophagitis and metaplasia have been
described in children following gastric tube replacement.
ere should be regular monitoring of the esophageal
conduit since chronic exposure to acid reflux may
predispose to metaplasia and adenocarcinoma.
1
Gastric transposition or the gastric pull-up procedure
has several advantages. It is relatively simple with a
single anastomosis at the neck, creating a suciently
long and very well-vascularized gra.
10,12
It has been
shown to have a reduced risk of anastomotic leakage,
stricture and necrosis.
5,8
e perioperative complications
in our study were esophageal leakage, esophagocutaneous
stula, delayed gastric function and dumping syndrome
(each arising in 33% of patients) and pneumonia (in all
patients). Vagotomy during gastric mobilization may
lead to delayed gastric emptying and dumping syndrome.
In addition, pyloromyotomy and pyloroplasty may be
performed to prevent delayed gastric emptying. Spitz et
al.
8,18
reported the outcomes of gastric transposition in 236
patients, which were a 2.5% mortality rate, a 12% leakage
rate and a 20% stricture rate. In our study, the long-term
outcomes of the gastric transposition group revealed
all patients (100%) achieved full oral feeding, with all
having anastomotic stenosis, oromotor dysfunction and
recurrent pneumonia. It is speculated that the recurrent
pneumonia was a consequence of underlying complicated,
type C, esophageal atresia. However, previous studies
have acknowledged that gastric transposition is associated
with a higher respiratory morbidity.
11
Occupying the
mediastinum and chest, the stomach may not empty
eectively, causing compression (mass eect) of the
intrathoracic organs and a long-term reduction of the
lung capacity. e long-term gastrointestinal outcomes
also included 33% gastroesophageal reux and dysphagia
rates. The reflux may lead to recurrent heart burn,
regurgitation, bad breath and pneumonia. Dysphagia has
been found to be common, despite successful and adequate
replacements
5,6
, which could be due to discoordinated
peristalsis, antiperistaltic layout, a tortuous esophageal
conduit or signicant acid reux.
Laohapensang et al.
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Original Article
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Colonic interposition is the most frequently used
esophageal replacement procedure and provides superior
length.
1,5
e perioperative complications in our study
showed 33% respiratory complication and gut obstruction
rates. Although there was a 100% wound infection rate,
there were no leakages or stulas. In the long-term
follow-up, all patients (100%) achieved full oral feeding,
and they developed dilatable anastomotic stenosis and
redundancy of the colonic gra; however, the patients did
not require intervention. e long-term gastrointestinal
outcomes comprised dysphagia (67%) and reux (33%),
while the respiratory problem was restrictive lung disease
(33%). In 2015, Lobeck et al.
14
reported that the most
common postoperative complication among 10 patients
after colonic interposition was esophageal stricture
(54%). e disadvantage of colonic interposition is the
redundancy of gras with stasis and dysphagia due to
a negative pressure in the thoracic cavity and emptying
by gravity.
4,5,19, 20
Complications such as anastomotic
leakage and strictures have also been reported to be
related to a precarious blood supply.
4
Other renowned,
large-scale series
8,15,18
with long-term outcomes have
demonstrated no signicant dierences in the early
or late complications of the gastric transposition and
colonic interposition techniques.
Jejunal interposition can be used as a pedicle or free
gra.
11
e main advantage of this gra type is the most
suitable caliber of the gras with peristaltic activity.
3
Conversely, high failure, morbidity and mortality rates
13
have been reported for this procedure, presumably due
to technical diculties and a tenuous blood supply. Our
center has no experience with this technique.
The average follow-up duration in the current
study was 3.6 years (range: 0.4-8 years). e measured
growth in patients aer esophageal replacement at the
nal follow-up demonstrated growth retardation and
malnutrition in all procedural groups, with 57% in the
isoperistaltic gastric tube, 100% in the reversed gastric
tube, 67% in the gastric transposition and 33% in the
colonic interposition groups. Malnutrition was deemed
to be present when a child’s weight-for-age fell below
the third centile of the standard growth chart. Growth
retardation is prevalent in patients with esophageal
atresia. Oral aversion is common among infants with
long-gap esophageal atresia due to delayed oral feeding,
caused by either a lack of sham feeding in oral feeds or
delays in the replacement.
5
It is important to monitor
nutritional status, growth and development in children
aer an esophageal replacement as they are known to
fall below centiles for both height and weight.
1,3,4
e limitations of a retrospective design, the small
number of patients and technical variations led to diculty
in making comparisons to discover any statistical signicance
in this study. Close monitoring and long-term follow-up
through more substantial group studies may elicit the
clinically important factors relating to the care of these
patients.
CONCLUSION
Esophageal substitution remains a major challenge in
children. Our experience indicates that children can be safely
operated on using any of the various methods currently
available, with acceptable perioperative morbidities and
no mortality. Nevertheless, the long-term consequences
and complications should be monitored throughout
adulthood.
Conict of interest: e authors have no conicts of
interest to declare.
REFERENCES
1. Borgnon J, Tounian P, Auber F, Larroquet M, Boeris Clemen
F, Girardet JP, et al. Esophageal replacement in children by
an isoperistaltic gastric tube: a 12-year experience. Pediatr
Surg Int. 2004;20:829-33.
2. Elshafei H, Elshafei E, ElDebeiky M, Hegazy N, Zaki A, Abdel
Hay S. Colonic conduit for esophageal replacement: long-term
endoscopic and histopathologic changes in colonic mucosa.
J Pediatr Surg. 2012;47:1658-61.
3. Gallo G, Zwaveling S, Van der Zee DC, Bax KN, de Langen
ZJ, Hulscher JB. A two-center comparative study of gastric
pull-up and jejunal interposition for long gap esophageal
atresia. J Pediatr Surg. 2015;50:535-9.
4. Lima M, Destro F, Cantone N, Ma M, Ruggeri G, Dòmini
R. Long-term follow-up aer esophageal replacement in children:
45-Year single-center experience. J Pediatr Surg. 2015;50:
1457-61.
5. Soccorso G, Parikh DH. Esophageal replacement in children:
Challenges and long-term outcomes. J Indian Assoc Pediatr
Surg. 2016;21:98-105.
6. Lee HQ, Hawley A, Doak J, Nightingale MG, Hutson JM. Long-
gap oesophageal atresia: comparison of delayed primary
anastomosis and oesophageal replacement with gastric tube.
J Pediatr Surg. 2014;49:1762-6.
7. Tannuri U, Maksoud-Filho JG, Tannuri AC, Andrade W,
Maksoud JG. Which is better for esophageal substitution in
children, esophagocoloplasty or gastric transposition? A 27-
year experience of a single center. J Pediatr Surg. 2007;42:500-4.
8. Spitz L. Esophageal replacement: Overcoming the need. J
Pediatr Surg. 2014;49:849-52.
9. Tabira Y, Sakaguchi T, Kuhara H, Teshima K, Tanaka M,
Kawasuji M. e width of a gastric tube has no impact on
outcome aer esophagectomy. Am J Surg. 2004;187:417-21.
10. Cowles RA, Coran AG. Gastric transposition in infants and
children. Pediatr Surg Int. 2010;26:1129-34.
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11. Reinberg O. Esophageal replacements in children. Ann N Y
Acad Sci. 2016;1381:104-12.
12. Ludman L, Spitz L. Quality of life aer gastric transposition
for oesophageal atresia. J Pediatr Surg. 2003;38:53-7.
13. Carraro EA, Muscarella P. Esophageal replacement for benign
disease. Tech Gastrointest Endosc. 2015;17:100-6.
14. Lobeck I, Dupree P, Stoops M, de Alarcon A, Rutter M, von
Allmen D. Interdisciplinary approach to esophageal replacement
and major airway reconstruction. J Pediatr Surg. 2016;51:
1106-9.
15. Burgos L, Barrena S, Andres AM, Martinez L, Hernandez F,
Olivares P, et al. Colonic interposition for esophageal replacement
in children remains a good choice: 33-year median follow-up
of 65 patients. J Pediatr Surg. 2010;45:341-5.
16. Gounot E, Borgnon J, Huet F, Sapin E. Isolated isoperistaltic
gastric tube interposition for esophageal replacement in children.
J Pediatr Surg. 2006;41:592-5.
17. Uchida Y, Tomonari K, Murakami S, Hadama T, Shibata O,
Shirabe J. Occurrence of peptic ulcer in the gastric tube used
for esophageal replacement in adults. Jpn J Surg. 1987;17:190-4.
18. Spitz L, Kiely E, Pierro A. Gastric transposition in children—a
21-year experience. J Pediatr Surg. 2004;39:276-81.
19. AbouZeid AA, Mohammad SA, Rawash LM, Radwan AB,
El-Asmar KM, El-Shafei E. e radiological assessment of
colonic replacement of the esophagus in children: A review
of 43 cases. Eur J Radiol. 2015;84:2625-32.
20. Vasseur Maurer S, Estremadoyro V, Reinberg O. Evaluation
of an antireux procedure for colonic interposition in pediatric
esophageal replacements. J Pediatr Surg. 2011;46:594-600.
Laohapensang et al.
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Review Article
SMJ
Ladawan Khowawisetsut, Ph.D.*, Narakorn Khunweeraphong, Ph.D.**
*Department of Parasitology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok 10700, ailand, **Medical University of Vienna,
Center for Medical Biochemistry, Max F. Perutz Laboratories, Campus Vienna Biocenter, Dr. Bohr-Gasse 9/2, A-1030 Vienna, Austria.
Extracellular Vesicles in Malaria Infection
Corresponding author: Ladawan Khowawisetsut
E-mail: ladawan.kho@mahidol.ac.th
Received 14 August 2017 Revised 12 January 2018 Accepted 15 January 2018
ORCID ID: 0000-0003-1585-495X
http://dx.doi.org/10.33192/Smj.2019.14
ABSTRACT
Malaria is one of the tropical diseases which cause high rate of morbidity and mortality. e disease is caused
by the infection of protozoan parasites in the genus Plasmodium. e severe syndromes of malaria infection arise
from the complex sequences of parasite-host interactions. It starts with parasite invasion and followed by the rupture
of infected red blood cells causing the release of parasite products that activate the host immune response. During
the past decade, research on the functions of extracellular vesicles (EVs) in many diseases including malaria has
increased dramatically. is article reviews the role of EVs in malaria immunopathogenesis. Investigations into
modulators in immune response, ubiquitous mechanism for intercellular communication between parasite-parasite
and parasite-host, as well as its usefulness as the diagnostic biomarkers are highlighted.
Keywords: Extracellular vesicles; exosomes; Malaria; Plasmodium spp. (Siriraj Med J 2019;71: 89-94)
INTRODUCTION
Extracellular vesicles (EVs) are heterogeneous types
of small membrane-enclosed particles which originated
from many cell types and can be found in body uids
such as serum, plasma and cerebrospinal uids (CSF).
EVs released from the cells in physiological conditions
have signicant role in homeostasis. Its level tends to
increase under the pathological conditions. EVs transport
cellular components such as proteins, lipids and genetic
materials from the originating cells to the recipient cells.
ese EVs can be directly fused at plasma membrane or
internalized by endocytosis resulting in the EVs cargo
transfer and function in the recipient cells. Nowadays,
the roles of EVs have been widely demonstrated in
many diseases such as cardiovascular diseases, cancer
and autoimmune diseases.
1,2
e contribution of EVs
is also well-established in infectious diseases. However,
study of EVs in the context of a parasitic infection is
complicated because both the parasites and the host
cells release the EVs into the extracellular space which
play a role in disease pathogenesis. e EVs in niche
environment provide the intercellular communication
between parasites and other parasites or host cells. It leads
to either activation or modulation in the host immune
response to parasites. is review provides an overview
of the research studies on EVs in malaria infection.
Characteristics of EVs in malaria infection
Although the denite terminology for dierent types
of EVs has not yet nalized. ree types of commonly
known EVs, apoptotic bodies, ectosomes and exosomes,
are classied based on their biogenesis and characteristics.
Because of their dierent biogenesis and originating
cells, each EV type contains distinct active biological
components and has specic biological functions.
3,4
e apoptosis bodies are the largest particles with
1-5 micron in size. ey result from outward blebbing of
plasma membrane of cells at the end stage of apoptosis.
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90
ey carry various molecules including the nuclear
fractions, cell organelles and genomic DNA which involve
in facilitating apoptotic process. e term of ectosomes, also
referred as microvesicles (MVs) or microparticles (MPs),
are used for dening the membrane-enclosed particles
ranging in size from 0.1 to 1 micron. MPs are released by
outward budding from plasma membrane of activated
cells or early apoptotic cells. e plasma membrane is
composed of a bilayer of lipids, including phospholipid
and oily substances. Phospholipids normally enriched
at the outer leaet of plasma membrane are cationic
and neutral phospholipids such as sphingomyelin and
phosphatidylcholine (PC) while anionic phospholipids
located at the inner leaet are phosphatidylserine (PS),
phosphatidylinositol (PI) and phosphatidylethanolamine
(PE). Once cells are in the activated stage or undergo
apoptosis, there is the alteration of plasma membrane
asymmetry, externalization of anionic phospholipids and
vesiculation from the plasma membrane. erefore, the
types of ectosomes can be identied by specic surface
markers such as phosphatidylserine, integrins, selectins
and other compositions expressed on the membrane of
originating cells. e exosomes are the smallest particles
with the diameter 40-100 nm. ey are generated from
the endocytosis of plasma membrane into endosomes and
later are accumulated as vesicles in multivesicular bodies
(MVB). e fusion of MVB with the plasma membrane
allows the release of exosomes from the originating cells
to the extracellular space. e common proteins used
for identifying exosomes include tetraspanins (CD81,
CD63, CD9), otillin, constitutive heat shock protein 70
(Hsc70) and vesicle trafficking-related proteins (TSG101,
ALIX, and RAB proteins, syntenin-1).
5
Both ectosomes
and exosomes deliver numerous proteins (cytoplasmic
proteins, transmembrane proteins and membrane associated
proteins), lipid and nucleic acids (mRNA, miRNA and
other non-coding RNAs), similar to those expressed
in the parental cells, to the membrane or cytosol of the
target cells and function in the same way as parental
cells do.
e characteristics of EVs derived from plasma of
malaria-infected patients and malaria culture supernatant
have been determined. e biophysical analysis of EVs
derived from P. falciparum-infected red blood cells
(iRBCs) showed that the majority of EVs were 50-300 nm
diameter with single bilayer membrane.
6
e proteomic
analysis of EVs revealed that they contained both host
and parasite proteins. e MPs isolated from plasma
of malaria-infected patients contained host proteins
including complement-associated proteins, coagulation-
associated proteins and cytoskeletal proteins. Meanwhile,
major parasitic protein components were involved in
parasite invasion and parasite growth.
7
e EVs from
P. falciparum culture supernatant are enriched with red
blood cell lipid ras proteins and membrane-associated
parasite antigens, especially proteins associated with
red blood cell membranes and proteins involved in
parasite invasion.
8,9
ese abundant parasitic proteins
included the parasite proteins found in cytosol of iRBCs
and exported to red cell membrane such as Maurer’s
cles; the merozoite secretory proteins such as RhopH
protein complex (RhopH1, RhopH2 and RhopH3), the
RAP complex (RAP2 and RAP3), RALP1 and RON3;
the microneme resident proteins such as EBA-175 and
EBA-181; and the dense granule proteins such as HSP101,
PTEX150, EXP2, SBP1, RESA, and MAHRP1.
9
Not
only proteins, but other genetic materials including
genomic DNA(gDNA), functional mRNA, miRNA and
other small non-coding RNA have also been detected
in EVs.
6,8,10
e parasitic gDNA, human and parasitic
small RNA between 4-150 nucleotides were present in
EVs derived from P. falciparum-iRBCs, especially those
from ring-stage iRBCs.
6
e miRNA prole of plasma
EVs and peripheral blood cells under normal healthy
conditions showed that red blood cell contributed the
highest cellular miRNA to the blood and dierent cell
lineage expressing dierent patterns of miRNA. e
miR-451 and miR-150 established crucial function in
erythroid- and lymphoid dierentiation, respectively.
e level of miR-223 is abundant in granulocytes and
platelets.
11
e highest miRNAs expressed in plasma
EVs were miR-223, -484, -191, -146a, -16, -26a, -222,
-24, -126, and-32.
12
Since Plasmodium spp., do not have
a mechanism to produce miRNA, all miRNAs isolated
from malaria-infected red blood cells or from the plasma
of malaria-infected patients have been conrmed to be
of human origin.
13-15
e majority of miRNA found in
iRBCs-derived EVs were miR-451, let-7b and miR-106b.
10
However, the expression levels of miRNAs, miR-19b,
-4732, let-7a, -16, -183, -18a and 148b in iRBCs-derived
EVs were lower than those in uninfected RBCs-derived
EVs.
6
EVs as an amplier in the pathology of malaria infection
Human malaria is caused by the infection of intracellular
protozoan genus Plasmodium transmitted by Anopheles
mosquitoes. ere are ve species of Plasmodium which
are P. falciparum, P. vivax, P. malariae, P. ovale and P.
knowlesi. Among these, the P. falciparum and P. vivax
are highly prevalent in ailand and the Southeast Asian
region.
16
Malaria infection begins with the injection of
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sporozoites from infected mosquitoes during a blood
meal. Sporozoites invade hepatocytes and proliferate into
merozoites. It leads to rupture of infected hepatocytes.
e hepatic merozoites enter the blood circulation, invade
erythrocytes and begin the erythrocytic schizogony. e
clinical symptoms are associated with the invasion of
asexual erythrocytic-stage parasite to erythrocyte, and
the inappropriate immune response to iRBCs and diverse
parasite-derived products. e severe P. falciparum infection
results from the sequestration of iRBCs, leukocytes and
platelets to endothelial cells (ECs) within microvessels, the
excessive proinammatory cytokine production and the
severe red blood cell hemolysis. e parasite sequestration
leads to accumulate host cells within vasculature, injure
ECs and disrupt blood ow, causing tissue hypoxia and
lactic acidosis. e sequestration mechanisms contribute
to organ-specic syndromes such as cerebral malaria and
placental malaria.
17
Recent studies in both animal models
and malaria-infected patients suggest the signicant
contributions of EVs to severe malaria.
In rodent malaria model, the association of EVs and
malaria pathogenesis was initially studied in ATP-binding
cassette transporter (ABC) knockout mice. e ABC
is a cholesterol transporter involved in controlling the
outward translocation of PS at the plasma membrane.
18
e
deletion of this gene results in reducing the externalized
expression of PS on the cell surface and inhibit the MPs
production. e Plasmodium berghei strain ANKA (PbA)
infection in the ABCA1 knockout mice (ABCA
-/-
) showed
the decreasing of plasma tumor necrosis factor (TNF)
level and resistance to cerebral malaria. e MPs from
these mice had lower procoagulant activity than those
from wide type mice.
19
Treatment with pantethine, a
provitamin regulated lipid metabolism, to PbA-infected
mice also reduced the MPs production, decreased platelet
reactivity and impaired endothelial cell activation by MPs
resulting in prevention of cerebral malaria development.
20
By contrast, the adoptive transfer of MPs from PbA-
infected mice with the neurological symptoms led to
localize MPs in cerebral microvessels of PbA-infected
recipient mice. e transfer of endothelial cell-derived
MPs (EMPs) also induces the signs of pathologies in the
brain and lung of the recipient mice.
21
ese ndings
indicate that host cell-derived EVs have pathogenic
roles such as procoagulant activity and proinammatory
potentials in the pathogenesis of cerebral syndrome.
Accumulating evidence of host cell-derived MPs in
plasma of malaria-infected patients has also supported
the contention that the elevated levels and origin of MPs
are associated with the disease severity. e levels of red
blood cell-derived MPs (RMPs), platelet-derived MPs
(PMPs) and EMPs were markedly increased in severe
falciparum malaria patients. e level of RMPs were
signicantly elevated in patients with severe anemia while
the levels of EMPs and PMPs correlated to coma depth
and thrombocytopenia.
22-24
Similarly, the levels of MPs
derived from leukocytes, platelets, and erythrocytes were
signicantly increased and the level of PMPs correlated
with the presence of fever in acute P. vivax-infected
patients.
25
ese elevated levels of MPs were reduced to
normal level at the convalescent phase and the clearance
of parasitemia.
24,26
Although the precise mechanisms underlying the
induction of MPs production during the course of infection
are not completely understood, many factors have been
described. e CD40L-induced platelet apoptosis and
thrombocytopenia were associated with increasing plasma
PMPs in PbA-infected mice with severe syndrome.
27
e
exposure to febrile temperature led to the signicant
increases of PS expression on the surface of iRBCs,
particularly at the late schizont stage before the red blood
cell egress which was corresponding to releasing EVs
from iRBCs.
28
Moreover, the level of proinammatory
cytokine TNF was positively correlated with the level of
circulating plasma MPs in malaria-infected patients, thus
TNF might be another factor that induces the releasing
of MPs in malaria infection.
26
e major cell sources of EVs in malaria infected
patients were from platelets and red blood cells. e in
vitro study showed that infected red blood cells produced
more EVs per cell than uninfected cells.
8,24
e components
that were carried by EVs were associated in driven malaria
pathogenesis. e EVs derived from plasma of malaria-
infected patients and P. falciparum culture supernatant
contain the proteins which are involved in parasite
invasion and parasite growth. Therefore, these EVs
might play a role in facilitating red blood cell invasion by
merozoite during intraerythrocytic life cycle. e plasma
MPs from cerebral malaria infected mice, but not from
non-infected mice, carried proteins that were implicated
in molecular mechanisms relevant to cerebral malaria
pathogenesis, including endothelial activation.
29
In addition,
the reticulocyte-derived exosomes of non-lethal P. yoelii
17X-infected mice contained the parasite antigens. e
immunization of these puried exosomes induced the
specic immune response to P. yoelii infected red blood
cells.
30
e EVs also transferred the functional miRNAs,
especially miR-451, from red blood cells to endothelial
cells and targeted on the genes of proteins required for
barrier function leading to vascular alteration.
10
In addition to these biomolecules, hexanal compound
was also encompassed with EVs of iRBC, because the
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92
volatile organic compound presented in EV of iRBC
is diacetin which is the insect attractant like hexanal.
However, both compounds have been proven to play a
role in malaria transmission.
31
EVs as a messenger in parasite communication
Within the past decade, EVs have emerged as important
mediators of communication between EVs-secreting cells
and recipient cells. Two crucial experimental studies
demonstrated the crosstalk between intraerythrocytic
stage -P. falciparum by EVs. Regev-Rudzki et al exhibited
the role of EVs cargo in parasite crosstalk by using the
transgenic parasites. e EVs derived from drug resistance
P. falciparum-iRBCs can transfer DNA encoding for a drug
resistance marker between individual parasites leading to
spreading of drug resistance in the parasite population.
Moreover, this study showed that EVs transferring between
P. falciparum parasites under drug pressure-induced
stress conditions allows increased dierentiation of
gametocytes.
32
In parallel, Mantel et al also revealed that iRBCs
are able to internalize EVs isolated from malarial culture
supernatant and transfer them into the parasite cytosol
leading to increased gametogenesis.
8
However, the exact
underlying mechanism of this phenomenon was not
obviously proven in this work. However, another study
showed that the endogenous translocation of human
miRNA-451 from red blood cells into the parasites leads
to chimeric fusion RNAs with regulatory subunit of
cAMP-dependent protein kinase (PKA-R) transcripts of
P. falciparum resulting in reduction of the translation of
the regulatory PKA subunit. e suppression of PKA-R
is associated with reduced parasite growth and increased
numbers of gametocytes.
33
All these ndings suggested
that the EVs transfer genetic materials of parasite and
host cells to parasites of other infected red blood cells
resulting in alteration of the parasite cycle.
EVs as a modulator on the host immune response
Internalization of EVs by immune cells leads to
either activation or suppression of the immune cell
function. Because monocytes are the key immune cells
that play a role in phagocytic eradication of iRBCs and free
merozoites in blood circulation during intraerythrocytic
life cycle, there are several interesting studies in monocyte/
macrophage immunomodulation by EVs during malaria
infection. e iRBCs-derived MPs from the plasma of
PbA-infected mice induce macrophage activation by
up-regulation of CD40 expression and proinammatory
cytokine TNF production. e activation is via TLR-4
and MyD88 dependent pathways resulting in systemic
inflammation which impacts on both progression
of disease complications and generation of adaptive
immune responses.
34
In addition, the internalization
of iRBCs-derived MVs isolated from malarial culture
supernatant by macrophages triggers the strong pro-
and anti-inammatory cytokine responses of TNF and
IL-10 production, respectively. e pre-incubation of
these MVs with neutrophils also reduces neutrophil
function.
8
Not only in malaria infection, the malaria
EVs also attenuate neutrophil function in response to
bacterial infection by inhibiting ability to produce reactive
oxygen species and suppression of cytokine secretion.
35
ese data demonstrate that EVs from iRBCs, but not
from uninfected red blood cells, strongly modulate the
cells of the innate immune system.
Immunization with reticulocyte-derived EVs
from P. yoelii 17X-infected mice can induce the
P. yoelii infected red blood cells-specic IgG antibody
production.
30
Study showed that the immunization with
the combination of these reticulocyte-derived EVs and
CpG-ODN to BALB/c mice leads to signicant increase
in the percentage of eector T cells of both CD4 and CD8
T cells, especially eector memory CD4 subset when
compared to mice immunized with reticulocyte-derived
EVs isolated from uninfected mice. In addition, in vitro
experiment showed that the exosome isolated from
P. vivax-infected patients are captured by splenocytes
leading to signicant increase of the number of CD3 T
cells and CD8 T cells, but there is no change on B or
NK cell population. ese data suggested that EVs also
activate the immune cells of the adaptive immunity.
36
In addition to the immunization of iRBCs-derived EVs
in malaria infection, the subcutaneous immunization
with the exosomes from excretory/secretory products
of Echinostoma caproni, an experimental intestinal
helminth, in mice can reduce symptom severity during
infection.
37
e intraperitoneal immunization of mice
with EVs isolated from Heligmosomoides polygyrus, a
gastrointestinal nematode, in alum adjuvant Is resulted
in induction of specic antibody response against larval
challenge and reduction of intestinal worm burdens.
38
Collectively, the parasitic EVs can also modulate diverse
aspects of the immune system, suggesting that these
EVs might be the candidate strategy used for vaccine
development.
EVs as a source of diagnostic biomarkers
Today, the circulating miRNAs have been the research
subject of interest as they can be used as biomarkers in
many diseases. Alteration of miRNA expression reects
the pathological status. Upon malaria infection, infected
Khowawisetsut et al.
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SMJ
hepatocytes or other tissues at pathological sites, such
as brain, placenta and bone marrow, may produce and
release the EVs containing tissue-specific miRNAs
to blood circulation. Detection of such miRNAs may
allow discrimination between infected individuals with
uncomplicated symptoms and those with specic organ
complications or to be used as the surrogate markers for
detecting the hypnozoites that remain dormant in the
liver.
e dierential miRNA expression study in the brain
tissues from PbA-infected mice showed that the levels of
miR-27a, miR-150, and let-7i, miRNAs in regulation of
cellular proliferation and the innate immune response, are
upregulated in infected mice with cerebral malaria when
compared to those without cerebral malaria symptoms.
39
e development of protective immunity against malarial
blood stages of P. chabaudi or the lethal outcome of
P. chabaudi infection in mice are also associated with
alteration of miRNA expressions in the liver during the
infections.
40,41
As human miRNAs, mir-451 and miR-16,
are highly expressed in red blood cells, it is not surprising
that these miRNAs are also detected in plasma of both
normal healthy and malaria-infected patients.
14,15
However,
their levels were signicantly downregulated in P. vivax
infection and negatively correlated with the severity of
parasitemia.
42
is might be due to the consumption of
miRNAs by parasites during parasite growth inside the
red blood cells and the clearance of circulating miRNAs
by spleen during infection. A recent study of the EVs-
bound miRNAs from plasma of mothers with placental
malaria showed overexpression of placenta miRNA, a
miRNA in regulatory process during gestation, compared
with non-infected group.
43
CONCLUSION
e involvement of EVs in the pathophysiology of
malaria infection have been extensively shown in both
malaria-infected patients and in experimental animal
studies. EVs are considered as a vehicle necessary for
transferring of proteins and nucleic acids which trigger
intercellular communication between parasites and
host immune cells which lead to change in the parasite
biology and regulation of host immune responses. Due
to their association with the disease severity, they are
now being researched as potential biomarkers and for
their use in future vaccine development.
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