The 16th CU-TU MT Student's Research Conference

Comparison of COX1 gene sequences of Strongyloides stercoralis in Thailand with GenBank
database

Suluckkana Sriiamkul, Anatapong shinakod, Dujdow Songthamwat
Department of Medical technology, Faculty of Allied Health Sciences, Thammasat University

Strongyloides stercoralis is a parasitic nematode that causes strongyloidiasis It is estimated that
approximately 30 100 million people are infected worldwide There are many previously phylogenetic studies
based on COX1 gene of S stercoralis but there has been no research about correlation of S stercoralis groups
divided by phylogenetic tree of COX1 gene and patient data in Thailand This study aimed to classify COX1
gene sequences of S stercoralis isolated from Thai patients in Vajira Hospital with similar sequences in
GenBank database by phylogenetic tree and analyzed their laboratory results and clinical symptoms The COX1
gene sequences of S stercoralis from 25 patients were analyzed by BLASTN to find similar reference
sequences in GenBank 95 identity Phylogenetic trees based on both COX1 genes of S
stercoralis sequences were constructed Our results showed that the phylogenetic tree based on Maximum
likelihood method resembles Neighbor joining method There was 20 clades including 12 clades that had both
sequences from our isolates and GenBank database Our isolates shared the same clades with those from
Myanmar, Lao People's Democratic Republic Laos , and Japan In addition, 40 of our patients were
immunocompromised and anemic Clade III, V, XII, XIV, XVII, XX Sixty eight percent had neutrophilia Clade
II, III, V, IV, IX, X, XII, XIII, XV, XIV, XVII, XVIII, XX and 24 had eosinophilia Clade II, III, IV, XIV, XIII In
conclusion, COX1 gene sequences of S stercoralis from 25 patients resembles GenBank sequences in the
same region The patient s laboratory results and clinical symptoms in each clade are preliminary information
for epidemiology However, the numbers of subject 25 samples are a limitation of this study More samples
should be performed

Keywords Strongyloides stercoralis, COX1 gene, Phylogenetic tree, Maximum likelihood, Neighbor joining

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Analysis of protein-protein interaction network of malaria parasite during
intraerythrocytic life cycle

Chookiat Zatang, Sirapat Pornprasittikul and Onguma Natalang
Department of Medical Technology, Faculty of Allied Health Sciences, Thammasat University

Malaria is a life-threatening disease. In 2019, there were an estimated 229 million cases of malaria
worldwide. The estimated number of malaria deaths stood at 409,000 in 2019. Children aged under 5 years
accounted for 67% (274,000) of all malaria deaths worldwide. In 2019, the African Region was home to 94% of
malaria cases and deaths.To study the interaction network of proteins of malaria in red blood cells by using
STRING program. Gather information to make a database of Proteins involved in the study of protein-protein
interaction networks were analyzed using STRING v11.5 program. interaction network group of proteins and a
table showing the relative values of each protein in that group. The results showed that Ring stage has a large
group of protein genes: RNA binding protein, Ribosomal protein, Ring stage protein, Merozoite stage has a
high expression protein gene group: Merozoite surface protein, Endoplasmic reticulum transport protein,
Trophozoite stage. High-expressing protein gene groups: Ribosomal protein, RNA binding protein, Schizont
stage; High-expressing protein gene group: DNA Replication Protein, Proteasome Protein. The results revealed
which protein gene groups were associated with different stages in erythrocytes of Plasmodium falciparum,
which could be used to study the response mechanisms of malaria parasites in erythrocytes and can be used
for the development of vaccines or antimalarial drugs to be more effective. The studied data can be gathered
in the database to be used as a basic knowledge in the study of the biology of malaria.

Keywords: STRING, Plasmodium falciparum, Ring stage, Trophozoite stage, Schizont stage

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Detecting SARS-CoV-2 and Bacteria on Highly Contaminated Surfaces at Thammasat
University Rangsit Campus and Nearby Areas

1Pakkaporn Archawametheekul, 1Sunisata Angkham, 1Nattamon Niyomdecha and 2Pirom Noisumdaeng
1Department of Medical Technology, Faculty of Allied Health Science, Thammasat University
2Faculty of Public Health, Thammasat University

The Novel Coronavirus 2019 (COVID-19) is an emerging pandemic disease caused by severe acute
respiratory syndrome coronavirus-2 (SARS-CoV-2), a current major public health problem internationally. SARS-
CoV-2 may survive on surfaces in public areas for considerable amounts of time, resulting in a risk of
transmission and infection. To eliminate virus contaminant, diverse forms of 70% isopropyl alcohol have been
widely used. However, negative impacts of continued copious use of alcohol during the COVID-19 pandemic
may incite bacterial alcohol resistance evolution. This study aimed to detect SARS-CoV-2 and bacteria on highly
contaminated surfaces on the Thammasat University Rangsit campus and nearby areas, and evaluate alcohol
susceptibility of sampled bacteria isolates. Fifty pooled swab samples were collected from specific
environmental surfaces. Each sample was divided into two parts to test for SARS-CoV-2 and bacteria,
respectively. The target gene of SARS-CoV-2 was detected by a real-time polymerase chain reaction (RT-
qPCR), while bacterial isolation and identification were done by culture and biochemical testing. An alcohol
susceptibility test with three different measures (70% ethanol, 80% ethanol and 75% isopropanol) was
subsequently performed and compared among isolated bacteria from this study, standard strains (ATCC), and
isolated bacteria obtained from before the COVID-19 pandemic. Results were that SARS-CoV-2 was detected
at 8% (4/50) from hospital restroom doorknobs and faucets, as well as on ATM keypads at the Talaad Thai fresh
market. In addition, up to 96% (48/50) of bacterial contaminants were detected in 25 medically important
isolates, categorized into three groups: Staphylococcus, Enterobacterales and non-fermentative bacteria.
Comparing alcohol susceptibility, 75% isopropanol was the most effective in destroying all groups of bacteria.
Notably, isolated bacteria from this study were significantly less susceptible to all types of alcohol tested, and
Staphylococcus was the most resistant to destruction.

Keywords SARS-CoV-2, COVID-19, Bacteria, highly contaminated surfaces, Alcohol susceptibility test.

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Investigation of 12 novel compounds for the antimicrobial effect on Streptococcus suis

Atitsara Kanjanawiwin, Merisa Khawwises, and Potjanee Srimanote
Department of Medical Technology, Faculty of Allied Health Sciences, Thammasat University

Although, Streptococcus suis isolated from patients in Thailand still have a satisfactory level of drug
susceptibility to the mainstay antibiotic, penicillin, the overall antimicrobial susceptibility pattern indicated the
tendency toward the intermediate susceptible and resistance in many antibiotics. In particular, the antibiotic
resistance of S. suis isolated from asymptomatic infected pigs that have the potential to transfer the drug
resistance S. suis to humans via the food chain, leading to the concern on antibiotic resistance S. suis infection
in humans in the future. Therefore, this study aims to investigate further the 12 novel compounds that can inhibit
the growth of S. suis effectively. The information on the structure of 12 novel compounds were revealed.
Bioactivities such as MIC value, putative target molecules, and mechanism of action were elucidated by using
the chEMBL web application. The S. suis MIC data of the 12 novel compounds were compared with that of the
relative Streptococcus species or other bacteria that had available data on those web applications. In case the
target molecule of a novel compound in other bacteria is known, the presence, DNA homology, and protein
similarity of that target molecule in the S. suis were elucidated using the BLAST search tool. The result of this
study shows that some novel compounds that can inhibit S. suis might have a target molecule similar to other
bacteria. In conclusion, chEMBL and BLAST search tool web applications are useful in silico analysis to predict
the antimicrobial activity and drug target in S. suis.
Keywords: Streptococcus suis, Gram-positive bacteria, Novel compounds, MIC, Target molecules

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Plasmodium knowlesi genetic diversity and susceptibility of human infection. A systematic review.

Bustan Samadeng1 , Naruesorn Mee-im1 and Jiraporn Kuesap2
1 Department of Medical technology, Faculty of Allied Health Sciences, Thammasat University
2Graduate Program in Biomedical Sciences, Faculty of Allied Health Sciences, Thammasat University

Plasmodium knowlesi is a naturally occurring malaria parasite in monkeys however naturally human
infections are reported with potential cause of severe and fatal especially in Southeast Asia. The prevalence of
infection is different in each endemic area. The epidemiological diversity may be related to the genetic
polymorphism of genes involved in human invasion. Therefore, this study was conducted to investigate the
genetic diversity of P. knowlesi gene involved in human invasion. A systematic review was performed by
retrieving published articles available from PubMed, Web of Science and Scopus. Titles, abstracts, and full-text
articles were screened by two independent investigators and assessed the quality of the selected research
articles using the JBI criteria. Out of the 3357 publications retrieved from the three databases, 27 publications
were eligible to be included in this systematic review. A total of eight genes were identified; P. knowlesi
circumsporozoite protein (PkCSP), P. knowlesi merozoite surface protein 1 (PkMSP-1), P. knowlesi merozoite
surface protein 3 (PkMSP-3), P. knowlesi apical membrane antigen-1 (PkAMA-1), P. knowlesi duffy binding
protein (PkDBP), erytrocyte binding protein, P. knowlesi normocyte binding proteins (Pknbp) and the
Plasmodium rhoptry-associated protein 1 (PkRAP-1) from Malaysia, Thailand and Singapore. The analysis of
nucleotide diversity, haplotype diversity, pairwise nucleotide different and phylogenetic revealed the diversity
of P. knowlesi invasion-related gene in each area, especially in Malaysia, P. knowlesi endemic areas. Therefore,
P. knowlesi invasion gene variants contribute to parasite endemic and virulence.

Keyword: malaria , Plasmodium knowlesi , genetic diversity , systematic review

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Study of mechanism of Ca mobilization between ER-mitochondria induced by nonstructural B
protein of enterovirus A

Utaiwan Taechaoatsawakun, Saranyata Suansiri and Jeeraphong Thanongsaksrikul
Department of Medical Technology, Faculty of Allied Health Sciences, Thammasat University

Nonstructural protein 2B of enterovirus A71 (EV-A71) has calcium (Ca2+) conductance activity. It has been
shown that EV-A71 2B protein could increase cytosolic Ca2+ levels and induce cell apoptosis in human SH-SY5Y
neuroblastoma cells. However, the mechanism has not been elucidated yet. In this study, the sources of Ca2+
mobilization in human embryonic kidney (293T HEK) cells transfected with EV-A71 2B protein were preliminarily
identified using Ca2+ channel inhibitors. BAPTA inhibits influx of extracellular Ca2+. BAPTA/AM is intracellular Ca2+
chelator. 2-APB, Felodipine, and Thapsigargin are Ca2+ channel inhibitors of ER that inhibit IP3R channel, RYRs
channel, and sarcoplasmic/endoplasmic reticulum Ca2+-ATPase (SERCA), respectively. Ruthenium red is Ca2+
channel inhibitor on mitochondria membrane namely mitochondrial uniporter (MUP). Intracellular Ca2+ can be
stained by green fluoresce dye Fluo-4/AM. It was found that 293T HEK untransfected control cells showed
homogenous pattern of Ca2+ staining. While the cells expressing EV-A71 2B protein showed discrete speckles of
Ca2+ staining. Upon treatment of Ca2+ channel inhibitors, the cells transfected with EV-A71 2B protein showed
homogenous pattern and reduced intensity of Ca2+ staining. We speculated the Ca2+ mobilization in human 293T
HEK cells transfected with EV-A71 2B protein is involved with Ca2+ channels on ER and mitochondria. The notion
warrants further experimental validations. The obtained data will facilitate identification of key molecules involving
Ca2+ dysregulation which mediated by EV-A71 2B protein. The gained knowledge will pave the way for drug
development for mitigate neuropathology caused by EV-A71 infection.

Keywords: Nonstructural 2B protein, Enterovirus A71, Human embryonic kidney (293T HEK) cell, apoptosis,
Ca2+ mobilization

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Development of falciparum K

Sirisopha Chandrasakha

Program of Medical Technology (or Department of Medical Technology), Faculty of Allied Health Sciences, Chulalongkorn
University

Malaria is an infectious disease caused by five Plasmodium spp. including P. falciparum, P. Vivax,
P. ovale, P.malariae, P.knowlesi. P.falciparum is significant species of malaria because it is the most frequent
species more virulent than other strains in Thailand. For malaria treatment, ACTs(artemisinin-based combination
therapy) is the first-line drug used for treatment of P. falciparum in Thailand but nowadays the P.falciparum
parasites are resistant to artemisinin caused by the mutations in the Kelch13 gene. Detection of Kelch13 gene
mutation is needed for malaria treatment guidelines.In this study, we aim to develop the droplet digital PCR

(ddPCR) assay for Kelch13 C580Y mutation. the ddPCR is a new PCR method utilizing a water-oil emulsion
droplet system. It can reduce turnaround time and increase sensitivity to detect gene mutation. The ddPCR

reaction was prepared in total 20 l per reaction containing the ddPCR reagents, forward and reverse primers,
probe, and the DNA template. The preliminary results of ddPCR condition show that an optimized PCR
annealing temperature of the assay was
droplets. Further study, we will use the novel developed ddPCR conditions for kelch13 mutation detection.

Keywords: Plasmodium falciparum Kelch mutations, Artemisnin-based combination therapy , Droplet digital
PCR (ddPCR).

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A survey of Toxoplasma gondii oocyst and other intestinal parasites from cat s feces in Bang
Sai Kai community and nearby areas

Sutima Chaikhwao1, Naruemon Klaharn1, Panyaporn Boonjitbanjong1
1Program of Medical Technology or Department of Medical Technology , Faculty of Science and technology,
Bansomdejchaopraya Rajabhat University

Toxoplasma gondii is a parasite of the Coccidia with cat as an important definitive host Man is
accidental host who may get infection in many ways such as eating food or drinking water with T gondii oocyst
contaminated from cat s faeces including contamination from hand to mouth ; eating undercooked meat with
T gondii cyst which cause Toxoplasmosis The disease is an opportunistic infection where a person who has
immune compromised will develop severe symptoms of the disease and may be fatal It is also a disease that
is dangerous for the fetus as it can be transplacental transmission and even an accidental infection in the
laboratory In Thailand, there are several reports of cat playing a key role in transmitting the disease to humans
In addition, there are other intestinal parasites of cat such as roundworms Toxocara cati , hookworms
Ancylostoma ceylanicum, Ancylostoma braziliense , tapeworms Taenia spp , Dipylidium caninum , liver fluke
Opisthorchis viverrini , where cat may be a source of disease transmission to humans Cats are the animal that
are very close to us both as pets or even stray cats which are often run around our places So they may be the
important source to transmit those parasites to us This bring us to this survey which have objectives to examine
100 cats faeces both pet and stray cats in Bang Sai Kai community and nearby areas for T gondii oocyst and
the other intestinal parasites by Simple smear, Formalin ether and Sugar floatation methods Till now a total of
35 fecal samples were examined from 11 stray cats and 24 pet cats The parasites are detected from 5 pet cats
which are T cati 2 cases and hookworm 3 cases and from 7 stray cats which are T cati 2 cases and hookworm
6 cases This preliminary results showed that both pet cats and stray cats in Bang Sai Kai and near by areas
have T cati and hookworm, the other parasites not found yet Stray cats have tendency to be more parasites
than pet cats In pet cats where the parasites are detected, they may not get drug treatment for intestinal
parasites or the owner did not take care them well In the stray cats where the parasites are detected, they may
run around in the environment to get food by themselves Those food may have more chance to contaminate
with the infective stage of the parasites to infect them

Keyword Toxoplasma gondii, Intestinal parasite of Cat, Stray cat, Pet cat

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Molecular Identification of Carotenoid-producing yeasts; Rhodotorula mucilaginosa

Laongdaw Nasom1, Kornkanok Thongdee1, Tawanwat Damnak2, Thanwa Wongsuk2, Potjaman Pumeesat1
1Department of Medical Technology, Faculty of Science and Technology, Bansomdejchaopraya Rajabhat University.
2 Department of Clinical Pathology, Faculty of Medicine Vajira Hospital, Navamindradhiraj University

Rhodotorula species are pigmented yeasts that are normally found in natural environments such as
soil, freshwater, seawater, plants and household appliances. Rhodotorula spp produce carotenoids exhibiting
yellow, red or orange colony characteristics. They have been considered as non-pathogenic fungi, and there
have been several previous case reports of Rhodotorula spp infection. Rhodotorula mucilaginosa, Rhodotorula
glutinis and Rhodotorula minuta are the most causative agents of human infection in this genus, especially
immunocompromised hosts such as hematologic malignancies, organ or bone marrow transplantation,
neutropenia and acquired immunodeficiency syndrome (AIDS). Besides patients with immune deficiency, the
presence of a central venous catheter (CVC) is an important risk factor of Rhodotoular infection. R.
mucilaginosa, an important emerging fungal pathogen, is the most prevalent species of Rhodotorula fungemia.
Thus, the aim of the current study was to identify R. mucilaginosa from carotenoid-producing yeasts isolated
from laundry machine samples. A total of thirty-two red/orange yeasts were grown on sabouraud dextrose agar
(SDA) and incubated at 25°C and 37°C for 24 and 48 h. DNA extraction was performed by boiling yeasts in
20 mM NaOH at 95°C for 45 min. Polymerase chain reactions (PCR) with fungus-specific primer of internal
transcribe spacer (ITS) 1-5.8S-ITS2 regions were amplified. PCR products were sequenced, edited, and
compared with existing data in the GenBank. The results demonstrated that all thirty-two carotenoid producing
isolates (100%) grew well and showed round, smooth, moist to mucoid, coral red to salmon-colored appearance
when grown at 25°C for 24 h. On the other hand, six isolates (18.75%) grew slowly at 37°C. They showed pin-
point colonies that were pale or did not show carotenoid pigmented type. Additionally, the size of PCR products
were approximately 600 base pairs (bp), and twenty-five R. mucilaginosa (78.13%) were identified by ITS1-
5.8S-ITS2 sequencing. The virulence factors of these yeasts should be investigated, particularly in terms of
antifungal susceptibility testing, biofilm formation, protease and phospholipase production, etc.
Keywords: Carotenoid-producing yeasts, Internal transcribe spacer, Polymerase chain reaction

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Effect of Phyllanthus amarus Supplements on Interferon-Gamma Production by T Cell in Obese
Subjects

Ratchawan Sungsuwan1, Juthamas Maneela1, Supawan Sangthong1, Nattaphol Prakobkaew1, Orachorn Boonla2, Surachat Buddhisa1
1 Department of Medical Technology, Faculty of Allied Health Sciences, Burapha University
2 Department of Physical Therapy, Faculty of Allied Health Sciences, Burapha University

Obesity has emerged as one of the most serious public health concerns in the 21st century, with more than
1 billion adults overweight, at least 300 million of them clinically obese and is a major contributor to the global
burden of chronic disease and disability. It has been recently recognized that obesity associated with impaired T
cell functions, a key component of adaptive immune system and increase risk for infections and cancer compared
with non-obese patients. Phyllanthus amarus belongs to the family Euphorbiaceae has been used widely in various
traditional medicines to treat of various diseases. Moreover, a study has previously shown that Niranthin isolated
from Phyllanthus amarus has the ability to enhance the proliferation and interferon-gamma (IFN- ) production by T
cells in responses to Leishmania donovani infection in BALB/c mice. However, no studies have been reported on
the effect of Phyllanthus amarus on IFN- production by human T cell. This study was aimed to investigate the effect
of Phyllanthus amarus supplements on IFN- production by T cell in obese volunteers. The result shown that
significantly decreased IFN- production in subject with obesity compared to non-obese and overweight subjects.
While the level of IFN- after oral administration of crude of P. amarus (150 mg/day) for 8 weeks was not significantly
different compared to pre-treated. Suggesting that obesity lead to impaired T cell function and Phyllanthus amarus
capsule supplement was no effect on IFN- production by T cell. The information may lead to understanding the
effect of P. amarus on T cell functions and the development of novel treatment to the diseases caused by impairing
of T cell functions in obese patients.

Keywords: Obesity, T cell, Interferon-gamma, Phyllanthus amarus

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Prevalence of latent tuberculosis infection (LTBI) of foreign workers in Chonburi province

Kunthida Parisutsoontorn, Natchalita Wattano, Sasiwan Hansa, Kanokporn Srisucharitpanit
Department of Medical Technology. Faculty of Allied Health Sciences. Burapha University

Latent tuberculosis (Latent TB) is an asymptomatic tuberculosis infection. However, when immunity is
reduced, Mycobacterium tuberculosis in the lungs can proliferate and symptoms occur. Active TB is difficult and
takes a long time to treat. This study aimed to detect latent TB of foreign workers in Chonburi Province. Four hundred
blood samples of foreign workers from Burapha University Hospital were collected. The Interferon-gamma release
assays (IGRAs) were used to detect latent TB infection. The Interferon-gamma (IFN- ) released from T-cells were
quantitative determined to indicate a tuberculosis infection. The QFT-Plus Tube coated specific antigen of
M. tuberculosis (ESAT-6 and CFP-10) induced CD4+ and CD8+ T-cell to release IFN- into plasma. Then, IFN-
was measured by ELISA. The controls were acceptable, so analytical procedures were reliable. One specimen was
positive for latent TB infection, whereas nine were negative from a total of twenty-two specimens. However, the
sample analysis result consisted of twelve samples that could not be interpreted (Indeterminate). The possibility
causes may be from specimen as hemolysis and lipemic. Importantly, pre-analytical techniques were not suitable
such as blood collection procedure, plasma separation, poorly mixing of blood with antigen coating on the surface
of blood collection tubes. Therefore, the testing process must be adjusted further.
Keywords: Latent tuberculosis, Interferon-gamma release assays (IGRAs), QuantiFERON-TB Gold Plus, Foreign
workers

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Oxidized High-density Lipoprotein Concentration in Type 2 Diabetes Mellitus

Kunanya Homphan, Vanida Sukvijit, Raveenan Mingpakanee
Program of Medical Technology (or Department of Medical Technology), Faculty of Allied Health Sciences, Chulalongkorn
University

Background: Cardiovascular diseases (CVD) are the leading cause of global mortality and a major contributor to
disability. CVD risk factors such as obesity, hypertension, and dyslipidemia are common in patients with DM,
placing them at increased risk for cardiac events. Total cholesterol, high-density lipoprotein (HDL)-cholesterol (HDL-
C), LDL cholesterol (LDL-C), and triglyceride are the conventional lipid markers for CVD assessment. Multiple
mechanisms exist for HDL to prevent atherosclerosis. Oxidized HDL on the other hand, loses its functions and
becomes dysfunctional HDL. More than HDL-C level, oxidized HDL measurement appears to be linked to CVD
assessment.
Objective: The aim of this study was to compare the oxidized HDL concentrations in type 2 diabetes and non-
diabetic volunteers.
Methodology : Blood samples from 18 non-Type 2 diabetes and 22 Type 2 diabetes volunteers were collected after
12-hour fasting. HbA1C, fasting blood glucose, total cholesterol, triglyceride, LDL-C and HDL-C were measured by
automated analyzer. The oxidized HDL was isolated from serum by precipitation with a commercial kit (Abcam,
Human). The oxidized HDL concentration in each group was evaluated by sandwich ELISA (Abcam, Human) and
compared the difference by using Shapiro-Wilk SPSS version 22)
Result : There were no significant differences in cholesterol, triglyceride, LDL, HDL, and oxidized HDL between
Type 2 diabetes (165.71 107.40) and non-Type 2 diabetes volunteers (132.16+98.94) when using = 0.05
(p-value = 0.112
Conclusion: From our experiment, there were no significant difference in lipid profiles between non Type 2 diabetes
and Type 2 Diabetes groups. The limitation of this study was to small sample size. To achieve more reliable results
in the future, we recommend using more samples in each population.
Keyword: Cardiovascular diseases (CVD), Type 2 Diabetes Mellitus, oxidized HDL

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Antioxidant and anti-inflammatory activities of edible mushroom Astraeus asiaticus

Kantheera Tangwongsupang , Kantapat Simmalee , Sunita Nilkhet2, Panthakarn Rangsinth , Siriporn Chuchawankul
Department of Medical Technology, Faculty of Allied Health Sciences, Chulalongkorn University
Program of Clinical Biochemistry and Molecular Medicine, Department of Clinical Chemistry, Faculty of Allied Health Sciences,
Chulalongkorn University
Department of Transfusion Medicine and Clinical Microbiology, Faculty of Allied Health Sciences, Chulalongkorn University

Studies on edible mushrooms have reported their potential antioxidant and anti-inflammatory activities. However,
the antioxidant activity of Astraeus asiaticus (AA) is yet to be explored. The present study aims to analyze the antioxidant
and anti-inflammatory activity of hexane and ethanol crude extracts from AA. The radical scavenging activity was investigated

-azino-bis (3-ethylbenz-thiazoline-6-sulfonic acid) (ABTS) and 2,2-di-phenyl-1-picrylhydrazyl (DPPH) radical
scavenging capacity assay and were reported as ascorbic acid equivalents (AAE µg/mg). the hexane extract (1 mg/ml)
showed scavenging activity at 1.76 ± 0.71 AAE µg/mg and 4.79 ± 1.56 AAE µg/mg, whereas, the ethanol extract (1 mg/ml)
showed the scavenging activity at 17.34 ± 2.89 AAE µg/mg and 8.42 ± 1.54 AAE µg/mg, respectively. the total phenolic and
total flavonoid were determined by Folin-Ciocalteu reagent that reported as Gallic acid equivalents (GAE, µg/g) and
Aluminium chloride colorimetric assay that reported as Quercetin equivalents (QE, µg/g), respectively. The total phenolic
content in hexane and ethanol extract (1 mg/ml) were found to be 1.39 ±0.12 GAE µg/mg and 17.59 ± 2.79 GAE µg/mg,
respectively. Where-in the total flavonoid content in hexane and ethanol extract (1 mg/ml) were found to be 4.20 ± 5.32 QE
µg/mg and 5.00 ± 4.84 QE µg/mg, respectively. Chemical analysis revealed that the ethanolic extract showed higher
antioxidant activity as well as higher total phenolic content in comparison to hexane extract. Furthermore, the binding affinity
between the phytochemical compounds in AA and enzymes involved in inflammatory processes, such as COX and LOX
were also investigated using molecular docking. However, the underlying molecular mechanism by which AA mediates the
antioxidant and anti-inflammatory response should be unveiled and understood in depth for further therapeutic applications.

Keyword: Astraeus asiaticus, Total phenolic assay, Total flavonoid assay, Scavenging radical capacity assay, Molecular
docking

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Recombinase Polymerase Amplification of RhD subtype DEL

Prattana Khamthakhruea1, Pensurad Uipanit1, Patrawadee Pitakpolrat2, Pattarin Tangtanatakul1
1Department of Medical Technology, Faculty of Allied Health Sciences, Chulalongkorn University
2Department of Laboratory Medicine, Faculty of Medicine, Chulalongkorn University

DEL (c.1227G>A) is a Rhesus blood group variant, found in up to 70% of Thai blood donors. Due to the imitation
of serology technique, DEL is difficult to identify in a routine laboratory. Molecular techniques have become a
method of choice to identify DEL variants because of its short turnaround time, high sensitivity and accuracy.
With fast and easy steps, Recombinase polymerase amplification (RPA) technique is a convenient molecular
technique suitable for routine usage.
Here, we aim to detect DEL variants using RPA.
First, the DNA was extracted from DEL blood donors. All samples (n=6) were confirmed to be the DEL variant
by sequencing and absorption-elution method. DEL c.1227G>A were specifically amplified. The PCR products
were determined by agarose gel electrophoresis.
According to the objective of our study, this result can be further applied for DEL screening in routine or point-
of-care kits development which result in better transfusion processes.

Keywords: Recombinase polymerase amplification (RPA), Polymerase chain reaction (PCR), DEL blood group.

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Detection of predicted Coa and Cob antigens using molecular techniques

Palida Khieorat1, Sunisa Ounjai1, Kamphon Intharanut2, Oytip Nathalang2
1Department of Medical Technology, Faculty of Allied Health Sciences, Thammasat University
2Graduate Program, Faculty of Allied Health Sciences, Thammasat University

Two antithetical antigens, Coa and Cob of the Colton CO blood group system are clinically important
in transfusion medicine To date, the data of those antigen frequencies among Thai populations remains
unknown This study aimed to determine CO A and CO B allele frequencies in Thai blood donors using
polymerase chain reaction PCR based techniques to predict Coa and Cob antigens and compare them with
other populations previously reported DNA samples obtained from 200 and 200 blood donors from central and
southern Thailand were genotyped by DNA sequencing Altogether, 400 samples were sequenced to identify
CO A and CO B alleles Primers were designed using Primers3 and BLAST A fragment of 150 bp containing
SNP c 134C>T in the aquaporin 1 AQP1 gene was obtained Analyses of the sequencing results in all 400
samples were performed by the SnapGene software Regarding the sequencing results, samples that exhibit
the CO A allele revealed the presence of c 134C, while the presence of c 134T represented the CO B allele
The genotyping results among 200 samples 100 0 of central Thais were homozygous CO A CO A For
southern Thais, 198 were homozygous CO A CO A 99 0 , and 2 were heterozygous CO A CO B 1 0
Homozygous CO B CO B was not found in both groups This study was the first to determine CO A and CO B
genotypes in two Thai populations using DNA sequencing This finding would be helpful to predict
alloimmunization risks related to transfusion induced reactions However, further study with a larger sample size
is suggested

Keywords Colton blood group, allele, genotype, Thai blood donors

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Factors affecting continuous donations among Thammasat University Hospital donors

Wachirada Naiwijarn1, Worakamon Pimsiri1, Oytip Nathalang2 and Kamphon Intharanut2
1Department of Medical Technology, Faculty of Allied Health Sciences, Thammasat University
2Graduate Program in Biomedical Sciences, Faculty of Allied Health Sciences, Thammasat University

Blood donations are essential to the blood supply available for patients in life saving treatments

including surgeries, cancer treatment, chronic illnesses, and traumatic injuries The COVID pandemic has

had an enormous impact on the number of blood donations, eventually, Thammasat University Hospital TUH

has been also affected The aim of this study was to identify factors that were affected continuous donations,

and to develop one such tool for predictive of repeat donation among TUH donors A retrospective study for a

period of years to donations at TUH was done to identify factors affecting continuous donations

and develop the scoring tool, as well as internal validation was done prospectively Data including age, sex,

ABO grouping, Rh D typing, and collection site were obtained on their continuous donations, factors that would

influence this decision Additionally, the outcome is dichotomized as controls and cases based on the first time

and continuous donations Receiver operating characteristic ROC curve was used to obtain the cut off for

independent variable age , odds ratio was used to assign the score During the study period, out of

donations, met the inclusion criteria controls ; cases Factors that positively predicted

intention to return included male OR , CI ; P < ; age, more than year old

OR , CI ; P < ; Group AB OR , CI ;P

and onsite donation OR , CI ; P , and their were chosen to develop the score

The total score value of was chosen as the cut off for the two groups It predicted repeat donations with a

strength of sensitivity and specificity On internal validation, the score was observed to have an

accuracy of In conclusion, male, age more than year old, AB blood group and onsite donation

appear to positively influence to repeat donation The developed predictive score is a simple reliable and handy

tool with high accuracy

Keywords Blood donation, first time donation, continuous donation, predictive score

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An approach to blood utilization and blood preparation at Pakkred Hospital 3 years experience
2018 2020

Natnapa Kladkleeb, Parisara Chanchek and Pramote Sriwanitchchrak
Department of Medical technology, Faculty of Allied Health Sciences, Thammasat University

The number of requests for blood transfusion in patients in each ward is currently expanding, which
may be resulted in the shortage of blood supply and increase the workload of blood bank personnel The
implementation of three index blood utilization management including crossmatch to transfusion C T ratio ,
transfusion probability T and transfusion index Ti is suggested The objective of this study was to study an
approach to blood utilization in each ward at Pakkred Hospital and then use the results to develop guidelines
for blood preparation for the next time Three years during 2018 2020 retrospective data demographics and
blood requests of patients in each ward was collected Three index blood utilizations of C T ratio, T and Ti
were analyzed There were 4,009 patients on each ward and total blood requests were 7,612 units Only 2,714
patients with 4,994 units were transfused C T ratio 1 5, T 67 7, Ti 1 2 The highest blood requests
were patients in Inpatient Department C T ratio 1 4, T 84 3, Ti 1 5 , followed by patients in Emergency
Room C T ratio 1 3, T 86 7, Ti 1 8 , followed by patients in Labour Room C T ratio 8 7, T 8 1, Ti

0 1 , followed by patients in Outpatient Department C T ratio 1 4, T 52 6, Ti 1 2 and patients in
Operating Room C T ratio 1 2, T 83 3, Ti 1 7 respectively This retrospective study showed the over
blood ordering of the blood bank at Pakkred Hospital It found that blood preparation was prepared more than
necessary The appropriate blood utilization and blood preparation should be used to reduce resources and
the cost of blood preparation

Keywords blood transfusion, blood preparation, crossmatch to transfusion, transfusion probability, transfusion
index

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The optimization of nanoparticle encapsulation for drug delivery system

Sujitra Promla, Pasinee kongkavee and Ratchaneewan aunpad
Department of Medical technology, Faculty of Allied Health Sciences, Thammasat University

Background Antimicrobial peptide AMP is one of biomolecule that has gained interest in drug
development due to its ability to inhibit or kill the drug resistant pathogenic bacteria However, AMPs remain
chemically and biologically unstable, which is a major obstacle for their clinical application Increasing the
stability of AMP with drug delivery systems for peptide encapsulation is therefore critical This research has
developed a nano scale drug delivery system or nanoparticles synthesized from chitosan and dextran sulfate
which are polymer with biocompatible, biodegradable, and low toxicity By electrostatic interactions between
anionic of the dextran sulfate and cationic of the peptide and chitosan, the peptide will be encapsulated within
the chitosan dextran sulfate nanoparticles Objective To determine the optimal ratio of chitosan and dextran
sulfate in nanoparticles preparation for peptide encapsulation Method The Nanoparticles were synthesized by
a complex coacervation method The peptide was mixed with dextran sulfate and chitosan was then added to
dextran sulfate or dextran sulfate was added to chitosan in different ratios consisting of 5 3, 4 4, and 3 5 by
weight After that the size and zeta potential were analyzed with Zetasizer and the antibacterial activity of
nanoparticles in inhibiting or killing pathogenic bacteria, Staphylococcus epidermidis ATCC 39584, was
determined by agar diffusion method Results The reduced ratio of chitosan added to the increased ratio of
dextran sulfate was associated with a reduction in the size of the nanoparticles and the increased ratio of
dextran sulfate added to the reduced ratio of chitosan was associated with an increase in the size of the
nanoparticles In addition, the zeta potential was also related to the ratio of dextran sulfate and peptide as the
nanoparticle encapsulation of peptide in each different ratio had more positive in total charge Nanoparticle
synthesis at the ratio 3 5 of chitosan added to dextran sulfate was the optimal ratio due to the formation of
nanoparticles of similar size homogenous size distribution According to the antibacterial activity experiment,
the empty nanoparticles had no antimicrobial activity while the nanoparticles encapsulation of peptide showed
antimicrobial activity, but all ratios have no difference Discussion This research demonstrated the mode of
addition and ratios of polymers related to the size and zeta potential of nanoparticles The ratio of 3 5 is the
most optimal ratio for the synthesis of chitosan dextran sulfate nanoparticles However, experiment of the
peptide encapsulation and release is required The study on the antimicrobial activity by other methods should
be done to select the optimal ratio for AMP encapsulation and use for drug delivery systems in the future

Keywords drug delivery system, nanoparticles, chitosan, dextran sulfate, antimicrobial peptide

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Page 68

Development and evaluation of quality control material for hematocrit testing with hematocrit
centrifuge

Patcharawee Tusnaimatakul, , Pramote Sriwanitchrak

Department of Medical Technology, Faculty of Allied Health Sciences, Thammasat University

Quality control material for hematocrit testing with hematocrit centrifuge of primary health care
laboratories are important for patient care Nowadays, EDTA blood is used as a quality control sample It must
be stored at 4 °C and has a short shelf life So, it s a limitation of the test The objective of this research is
developed and evaluate of quality control material for hematocrit testing with hematocrit centrifuge From
processed blood for use in primary care laboratories for quality control Processing expired blood from a blood
bank with 2 5 glutaraldehyde to treat the condition After that, hematocrit volume was adjusted to three levels
low 21 , normal 42 , high 51 , and the percentage of hematocrit was measured by centrifugation In this
study, samples of all three quality control levels were tested for homogeneity In addition, stability was tested
for a period of 3 months The prepared quality control samples were kept at room temperature for the duration
of the research The results showed the target value of 100 percent the same accurate test results for the
homogeneity test and the stability test of quality control material for hematocrit The test results showed the
quality control material for hematocrit were suitable But Further research is needed to practice in the laboratory
for appropriateness for the quality control material for hematocrit testing

Keywords Hematocrit testing, Proficiency testing, Primary health care unit

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Page 69

Development of paper-based immunoassay with immunogold silver enhancement for oxidized LDL
detection

Dolaporn Pattarathitinuruk, Apichaya Mongkolvorawan, Raveenan Mingpakanee
Program of Medical Technology (or Department of Medical Technology), Faculty of Allied Health Sciences, Chulalongkorn
University

Cardiovascular disease (CVD) is one of the leading causes of death in the world. Total cholesterol,
triglycerides, high-density lipoprotein cholesterol (HDL-C), and low-density lipoprotein cholesterol (LDL-C) are all
lipid profiles used in routine laboratories to assess the risk of CVD. LDL-C is a major risk factor for cardiovascular
disease. LDL can be oxidized, resulting in oxidized LDL (ox-LDL). The ox-LDL was not taken up by the LDL receptor,
but it is taken up by macrophages via the scavenger receptor, resulting in the formation of foam cells in the arterial
wall. The LDL-C concentration in the routine laboratory is a direct enzymatic colorimetric method. This method can
only detect quantity, but It cannot differentiate between LDL and oxidized LDL.The oxidized LDL can be detected
by sandwich enzyme-linked immunosorbent assay, however, the enzymes have a limited shelf life and difficult to
maintain. The purpose of this study is to develop a paper-based immunogold silver enhancement assay.
Monoclonal anti-HNE antibodies are immobilized on Whatman paper and secondary antibody is anti-apoB
conjugated gold nanoparticles. The gold nanoparticles reduce silver ions into metallic silver in the presence of a
reducing agent. The reduced silver then deposits on the gold surface, thus enlarging the size of the gold
nanoparticles. The results can be measured using a desktop scanner and analyzed using ImageJ software and
scaled to have RGB value. The benefit of this technique is reducing the time and cost, no specific equipment is
required. The ox-LDL concentration could be a new CVD marker for CVD risk assessment.
Keywords: Oxidized LDL, Paper-based, Immunogold Silver Enhancement.

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Page 70

Antitumor activity of Pleurotus Sajor-Caju on triple negative breast cancer cell line

Nutchaworada Thitirattananon¹, Nathnaree Taebunpakul¹, Sunita Nilkhet², Panthakarn Rangsinth³, Siriporn Chuchawankul³
¹Department of Medical Technology, Faculty of Allied Health Sciences, Chulalongkorn University
²Program in Clinical Biochemistry and Molecular Medicine, Department of Clinical Chemistry, Faculty of Allied Health Sciences,
Chulalongkorn University
³Department of Transfusion Medicine and Clinical Microbiology, Faculty of Allied Health Sciences, Chulalongkorn University

Breast cancer is the most commonly diagnosed cancer in woman globally. Also, it is the main cause for
cancer related death. The high incidence of breast cancer is Triple negative breast cancer (TNBC), it defined by

. There are the reasons
that make TNBC has the worst prognosis and difficult to treat by hormonal therapies unlike the other breast cancer
types. In this study, we interested in Pleurotus Sajor-Caju (PS) or Oyster mushroom. It is a common culinary
mushroom found in Southeast Asian countries, including Thailand which is commercially relevant and high in
nutritional value. Further, PS extracts are known to show potential immunomodulatory and antitumor activities.
Previous studies on HCT116WT colorectal cancer cell lines revealed the induction of apoptosis by PS .Moreover,
anti-proliferative activity was shown against U937 leukemia cells. However, the antitumor activity of bioactive
compounds from PS against triple negative breast cancer (TNBC), MDA-MB-231 cell line have not been well
explored. Hence, in this study, we intend to investigate the antitumor activity of the PS maceration extract on MDA-
MB-231 cells and HaCaT cells as a representative of normal cells. MTT assay revealed that reduction of cell viability
in MDA-MB-231 cells in a dose dependent manner (6.25 to 100 µg/mL) in response to treatment with PS maceration
extract compared to untreated cells after 24 and 48 hrs compared to HaCaT cells. Though PS showed cytotoxic
effect on HaCaT cells, it exhibited anti-proliferative activity against MDA-MB-231, showing its potential as anticancer
therapeutic.

Keywords : Pleurotus sajor-caju (PS), Triple negative breast cancer (TNBC), MDA-MB-231 cells, MTT assay

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Page 71

The Immunomodulatory effect of Pleurotus sajor caju crude extract in human monocyte

Kulsiree Sutthirat, Fasai Jainuan, Kamolporn Amornsupak
Program of Medical Technology or Department of Medical Technology , Faculty of Allied Health Sciences, Chulalongkorn
University

Infectious diseases are disorders caused by pathogenic microorganisms such as bacteria, viruses,
fungi, or parasites Despite the availability of antibiotics and vaccines against causative pathogens, the mortality
rate remains high which is in part due to drug resistant infections This problem could be prevented by using
antibiotics wisely and well infection prevention and control The innate immune system plays an important
defense role against foreign invaders and protects the infection Therefore, factors that could enhance innate
immune response would be a benefit to control such diseases Pleurotus Sajor caju PS , a macro fungus,
contains high beta glucans which act as pathogen associated molecular patterns This study is aimed to study
the immunomodulatory effect of the PS crude extract in human monocyte using THP 1 cell line The appropriate
concentration of PS crude extract for THP 1 treatment was determined by MTT assay Then, THP 1 cells were
stimulated with PS crude extract for 24 h After that, the supernatant was collected and measured by ELISA
assay kit The increased proinflammatory cytokines collected from PS crude extract treated cells were analyzed
and compared with untreated control cells and PMA or beta glucan treated cells as negative control and
positive control, respectively The preliminary results obtained from this study will be the basic knowledge that
highlights the impact of PS crude extract as an immunomodulator or adjuvant for the prevention and treatment
of important pathogenic infections in the future
Keywords Pleurotus sajor caju, Immunomodulatory effect, monocyte, beta glucan

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Page 72

Characterization and limitation study of simultaneous paper-based Rh blood typing device

Chananchida Kamtan, Siwakorn Seedapalee, Tasanee Sakuldamrongpanich, Wanida Laiwattanapaisal
Program of Medical Technology (or Department of Medical Technology), Faculty of Allied Health Sciences, Chulalongkorn
University

The Rh blood group is regarded as one of the most vital blood groups in blood transfusion due to its highly
immunogenic resulting in severe transfusion reactions. Although a conventional tube is a gold standard method for
blood group typing, it requires large amount of antibody and its disadvantage in terms limited antibodies shelf-life,
Therefore, most laboratories of the General Hospitals did not spare the antibodies for Rh blood typing. In this
research, paper-based analytical devices (PAD) for simultaneous 5 Rh antigens typing, including D, C, E, c and e,
has been proposed for simple, low-cost and superior storage stability. The principle was based on the presence or
absence of hemagglutination at the PADs detection zones where the antibodies were pre-immobilized. The antigen-
positive RBCs reacted and agglutinated with their corresponding antibodies while the antigen-negative RBCs failed
to agglutinate and were eluted by the elution buffer, thus visualizing the red color at the channel. In the previous
study, the limitations of the proposed device have not been systematically investigated. To investigate the effect of
hematocrit, the EDTA whole blood with different percentage of hematocrit (%Hct) ranging from 10% to 60% were
tested. Anti-D and 1%BSA were immobilized and tested for the positive and negative hemagglutination,
respectively. The results show that, the method potentially be used in Hct ranges from 10-60%, as agglutination or
non-agglutination could be clearly observed. However, further study should be evaluated with the other Rh antigens.
Furthermore, studies on the stability of the device should be elucidated to increase the performance of paper-based
Rh blood typing device.
Keywords: Rh blood group, simultaneous paper-based blood typing, hemagglutination, whole blood

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Effect of Lysiphyllum strychnifolium Craib A Schmitz extract on reducing proinflammatory
cytokine in THP 1 cell line

Thatsawan prakobkhetkorn, Nichakorn kaengkarn, Metawee weruwanarug, Artittaya Rodchanasarod, Piya Wongyanin
Program of Medical technology Bansomdejchaopraya Rajabhat University

Lysiphyllum strychnifolium Craib A Schmitz have high level of phenolic compounds Its extract
showed high antioxidant and immunomodulation activity In this study, effect of Lysiphyllum strychnifolium
Craib A Schmitz extract on reducing quantity of proinflammatory cytokine was studied in THP cell line by
sandwich ELISA technique The result showed that, THP 1 cell line co cultured with Lysiphyllum strychnifolium
Craib A Schmitz extract at concentrations of 0 4, 0 2 and 0 1 g ml and LPS 200ng ml can reduce quantity of
proinflammatory cytokine namely IL 1, IL 6, IL 8 and TNF compared to control THP 1 cell line cultured with
LPS 200 ng ml only In conclusion, the result from this study indicated that extract from Lysiphyllum
strychnifolium Craib A Schmitz can reduce the quantity of proinflammatory cytokine so it can also be reduced
inflammation in cell Moreover, knowledge from this study will help to develop gauze pad from Lysiphyllum
strychnifolium Craib A Schmitz extract in the future
Key word Lysiphyllum strychnifolium Craib A Schmitz, proinflammatory cytokine, sandwich ELISA

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Page 74

Study on T follicular helper cell response to SARS-CoV-2 vaccination

Nunnaphat Mekjinda, Thanchanok Jenjitsiri, Pornpimon Angkasekwinai
Department of Medical Technology, Faculty of Allied Health Sciences, Thammasat University

Coronavirus disease 2019 (COVID-19), an infectious disease caused by the virus Severe Acute
Respiratory Syndrome-Coronavirus-2 is highly contagious and the outbreak quickly spreads around the world.
COVID-19 disease can be transmitted through the respiratory system, which causes fever, cough, tiredness,
and difficulty breathing and these symptoms could be severe, especially for elderly and people with
immunodeficiency, giving a rise to a higher mortality rate. Immunization studies against infection and COVID-
19 vaccination are extremely significant to people, since vaccination reduces the risks of SARS-CoV-2 infection.
There are different types of vaccines against COVID-19, including viral vector vaccine, inactivated vaccine, and
mRNA vaccine. Initially, inactivated vaccine, Sinovac was the main Covid-19 vaccine regimen in Thailand.
Different types of vaccination show variable efficacy of preventing infection and disease severity. Currently, the
level of immunity to SARS-CoV-2 after vaccination was measured based on the levels of antibodies with no
much considering on the induction of antigen specific T cell responses. The previous studies found that T
follicular helper cells (TFH) are responsible for the activation of B cell responses. The purpose of this research
aims to study SARS-CoV-2 specific T cell response and T follicular helper cells using peripheral blood
mononuclear cells (PBMC) from people who received 2 doses of Sinovac, Astrazeneca-Pfizer combination, and
2 doses of Astrazeneca. The Activation-induced marker (AIM) assay after stimulation with SARS-CoV-2 S1
antigen were used to analyzed SARS-CoV-2-specific CD4+ T and TFH cell response using flow cytometry
analysis. We found that the frequencies of SARS-CoV-2 specific T follicular helper cells in people who received:
Astrazeneca-Pfizer, Astrazeneca, and Sinovac were 4.29%, 1.4% and 1.20%, respectively. As a summary, this
research indicates the activation of T follicular helper cells in the immune response to the SARS-CoV-2 vaccine.

Keywords: T follicular helper cell, SARS-CoV-2, Activation induced cell marker cell (AIM assay), COVID-19
vaccine

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Page 75

Transcriptome profiling of macrophages response to Cryptococcus neoformans infection

Panyawadee Triphop, Jirachaya Hattachote, Pornpimon Angkasekwinai
Department of Medical technology, Faculty of Allied Health Sciences, Thammasat University

Cryptococcosis is caused by Cryptococcus neoformans, an opportunistic fungus that causes disease
in immunocompromised patients, especially in HIV-infected patients, whose mortality rate is often due to
pathogen transmitted to the brain. Previous studies have shown that the pathogen can infect macrophages and
transmit to the brain and leading to meningitis. Different macrophage responses can cause different disease
outcome. However, macrophages response to different stages of the disease after C. neoformans infection has
not been studied. We hypothesized that macrophages may have a response pattern that promotes the spread
of infection during the disease progression. Therefore, the aim of this research was to study the whole
transcriptome profile of macrophages responses during the progression of C. neoformans infection. We
employed mouse model of cryptococcosis. The alveolar macrophage was isolated at 7 and 14 days post-
infection using the Fluorescence-activated cell sorting (FACS) and the expression of genes at the transcriptome
level was studied using RNA sequencing. Venn diagram data presented 1 7 2 and 4 4 6 genes uniquely
expressed in macrophages infected for 7 and 14 days, respectively. For the hierarchical clustering analysis
carried out using expression level FPKM of differential expression genes revealed a distinctive expression
profile of macrophages response at 7 and 14 days after infection. Volcano plot analysis revealed that the
highest gene groups in the top 10 genes after day 7 infection were Serpina3 g, Iigp1 , Arg1 , Ifi4 4 , Tgtp2 ,
Gm4951, Nos2, Serpina3f, Tlr12 and Gpb11, whereas day 14 were Arg1, Nos2, H2-M2, Ccl24, Tmeff1, Iigp1,
Ddx3y, Retnla, Ccl8 and C1qa. Thus, our data suggest that macrophage phenotype after infection in the chronic
stage is different to the early stage by upregulating Arg1 , C1 qa, H2 - M2 and Ccl2 4 that were related to the
transformation of macrophage into an alternative activated macrophage (M2) phenotype. This research
provides understanding to the role of macrophage and can be used as a guide for further study of cell functions
in C. neoformans infectious disease.

Keywords: Cryptococcus neoformans, Cryptococcosis, Macrophage, RNA sequencing, Transcriptome profile

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Page 76

Delivery Route of Tumor Antigens in Dendritic Cells

Warintorn Thammathaporn, Phattawan Chaiya and Sawang Petvises
Department of Medical technology, Faculty of Allied Health Sciences, Thammasat University

Dendritic cells (DCs) are white blood cells that act as professional antigen presenting cells by bridging
between innate and adaptive immunity. DCs also induce tumor immune response by activation of anti-tumor T
cells. DCs can phagocytose tumor cells, process and deliver varieties of antigenic peptides to induce cytotoxic
T cell activity. This study has reviewed literature about delivery route of tumor antigens in dendritic cells to
optimize the immune response in cancer. The five major types of tumor antigens including of DNA, tumor cell
lysate, mRNA, protein and peptides, which were primarily searched from the Pubmed database, primarily
appeared in human experiment only, were selected for consideration. Selected original articles were assessed
and evaluated the application feasibility, safety and efficacy in inducing T cell immunity. According to the
relevant studies, tumor DNA responsible for the mutated cancer genes has a high ability to induce immune
response, but delivery of tumor DNA may pose a harm due to integration of tumor DNA to host genome. Tumor
cell lysate covers up all known and unknown tumor antigens that can activate T cells. However, it is hard to
determine which tumor antigens are capable of activating T cells and may induce autoimmune reactivity by
epitopes shared in normal tissues. Proteins are antigens present in all types of cancer. Tumor proteins cannot
be presented directly and therefore need to be processed into peptides before presenting onto the MHC
molecules. Tumor peptides can induce DCs directly via the MHC molecules, requiring minimal processing prior
to presentation. However, loading peptides requires HLA haplotypes identification. The mRNA delivery into DCs
facilitates the processing and presentation both class I and II MHC-restricted epitopes. Furthermore, mRNA
cannot integrate into the host genome, indicating the safety of mRNA usage. The mRNA has a short half-life
and can be degraded easily according to route of delivery and vector used. Based on assessment and
thoroughly review, we expect that tumor mRNAdelivery would be the best alternative for inducing tumor immune
response via dendritic cells.

Keywords: dendritic cell, tumor cell lysate, protein, peptide, mRNA

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Page 77

Proteomic analysis of plasma proteins in human immunodeficiency virus-infected patients under
antiretroviral therapy

Harit Saepim, Kanyaphat Charoenvorachai, Chareepporn Akekawatchai
Department of Medical technology, Faculthy of Allied Health Sciences, Thammasat University

Human immunodeficiency virus (HIV) is the cause of AIDS, remains a major public health problem
worldwide. HIV patients has been treated by using Anti-retroviral therapy (ART). results in a reduction in the
mortality rate of people living with HIV. But the study of people with HIV who were treated with ART was found
to have side effects after taking the drug for a long time. The infected people tend to develop non-AIDS co-
morbidities. The mechanism of this disease is caused by the activity of the immune system and the inflammatory
process. Therefore, this study aimed to examine changes in plasma protein profile involved in inflammatory
processes in response to antiretroviral therapy in HIV-infected patients by proteomics. We collect specimens
from a group of people infected with HIV. The experimental group was divided into two groups: 32 people with
HIV who had never received ART and 73 people who had received ART. The baseline clinical and laboratory
data were age, sex, CD4+ levels. Then the plasma was analyzed to compare protein by proteomics (Heat map,
Venn diagram and STITCH 5.0) to look for protein that may respond to antiretroviral therapy in people with HIV.
The result showed that the median age of participants was 40 years. From heat map result, the expression of
protein is diverse. Then, we put it in Venn diagram, it has 75 proteins that expression difference in HIV patients
treat with and without ART. We take 75 proteins to predict by STITCH 5.0. The candidate proteins were
associated with inflammatory markers and MTOR signaling pathway. Our data indicated that inflammatory
markers and MTOR signaling pathway may be protein involved in inflammatory processes in response to ART.

Keywords: anti retroviral therapy, Human immunodeficiency virus, proteins, inflammatory process

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Effects of long term ketogenic diets on cardiovascular risk in type 2 DM A systematic review

Siriyakorn Larbmak, Noppasorn Saripan and Kanyanath Piumngam
Department of Medical technology, Faculty of Allied Health Sciences, Thammasat University

The prevalence of type 2 diabetes mellitus T2DM has been rising more rapidly nowadays This
chronic disease causes chronic hyperglycemia and leads to damage to many systems of the body and many
complications over time Current treatment involves diet and exercise to lower blood sugar levels and reduce
other risks such as lipidemia control to prevent potential complications Some previous studies have suggested
that a ketogenic diet or low carb, high fat diet is effective for weight loss and glycemic control in T2DM
However, there is no adequate study data on the sustainability, safety, and long term efficacy of the ketogenic
diet in T2DM patients including the effect on cardiovascular risk, and long term results are controversial This
study aims to systematically review the effect of the long term ketogenic diet on the risk of cardiovascular
disease in patients with T2DM A systematic review was searched on 3 databases included Scopus, PubMed,
and Web of science The inclusion criteria were a clinical trial published in English Participants were T2DM
patients on the ketogenic diet for more than 6 months This study focused on the effects of the long term
ketogenic diet on cardiovascular risk factors JBI s critical appraisal tools were used to assess the quality of
published papers Eight studies met the inclusion criteria and there were 371 participants with T2DM on the
ketogenic diet followed from 6 months to 2 years Five studies measured blood pressure and all of them showed
a result in decreased Seven studies measured blood glucose levels, 2 of them had no significant increase in
blood glucose level and 5 studies had a decrease in blood glucose level, 4 out of 5 had a significant decrease
All the studies 8 studies measured the lipid profile and all of them had an increase in HDL cholesterol, 7 out
of 8 studies reported a decrease in total cholesterol, LDL cholesterol, and triglyceride This systematic review
can conclude that a long term ketogenic diet improves cardiovascular risk in patients with T2DM and is safe to
use in long term 2 years It may also be used to control the potential therapeutic role of the ketogenic diet in
the long term management of T2DM

Keywords Long term ketogenic diet, Type 2 diabetes, Cardiovascular risk

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Page 94

The suitable non-invasive markers for advanced hepatic fibrosis in type 2 diabetic patients

Kunthida Poungpae, Wirada Nudang and Pilaiwan Siripurkpong
Department of Medical technology, faculty of Allied Health Science, Thammasat University

Fatty liver and liver fibrosis are found in Type 2 diabetic patients (T2DM) more than non-diabetic
patients. Non-alcoholic fatty liver disease (NAFLD) in T2DM can cause the advance hepatic fibrosis (AF),
cirrhosis, and the mortality in T2DM, respectively. The gold standard technique for diagnostic of AF is liver
biopsy, which is an invasive method and are needed the specialized physicians for result interpretation.
Therefore, non-invasive markers are interested as a screening test for AF before liver biopsy is chosen. From
previous study, non-invasive markers with varying the cut-off value were suggested for diagnosis of liver fibrosis
in T2DM. However, the suitable non-invasive marker for AF in T2DM are remained unclearly. Therefore, this
study aimed to compare the efficiency of non-invasive markers and look for the suitable markers for diagnosis
of AF in T2DM. This research study is a systematic review using PubMed database during 2015-2021 with
search term based on PICO. The search strategy is included patients with type 2 diabetes, and FIB-4 and/or
APRI, and advanced hepatic fibrosis, and liver biopsy as reference method. After excluding unrelated papers,
only 4 relevant original papers were extracted for the cut-off value, the sensitivity and specificity of non-invasive
markers for advanced liver fibrosis in T2DM and T2DM with NAFLD. The results were as follows: for FIB-4, the
cut-off value >1.45 closely showed the sensitivity and specificity values, but the cut-off value >2.67 presented
a low sensitivity (22-44%), but high specificity (93-97%), for APRI, the cut-off value >0.42 showed a similarity
of sensitivity and specificity, but the cut-off value ≥ 1.00 had a low sensitivity (16-27%), and high specificity
(94-97%). In summary, FIB-4 cut-off >2.67 and APRI cut-off ≥ 1.00 are inappropriate as screening test for
advanced hepatic fibrosis in type 2 diabetic patients due to their low sensitivity. Patients with T2DM who are at
risk for liver fibrosis may be excluded from further diagnosis by liver biopsy.
Keywords: Type 2 diabetic patients, Advanced Liver/hepatic fibrosis, Non-invasive marker, FIB-4, APRI

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Page 95

Study of control material matrices containing red blood cells for finger-stick blood glucose meters

Natthinee Larpvorakitchai1 , Chorthip Charoensilp1 , Somsak Fongsupa1
1Department of Medical Technology, Faculty of Allied Health Sciences, Thammasat University

Finger-stick blood glucose meters have an increasing role in glycemic control for the prevention of diabetic
control materials with the same matrices as the finger-stick blood sample are not commercially available. The
artificial glucose control material containing red blood cells in 0.85% NaCl was developed and can only be used
with some finger-stick blood glucose meters. It is not still able to use for approved glucometers in Thailand. This
study aimed to demonstrate the effects of the matrices of glucose control material on glucose determination by
finger-stick blood glucose meters. Three levels of glucose control materials (low, medium and high) containing
glutaraldehyde-fixed red blood cells were prepared in 0.85 % sodium chloride solution (NSS) and saline-citrate
buffer (pH 7.4), and then determined by automatic chemistry analyzer and five finger-stick blood glucose meters
including Gluco-Navii, Contour Plus, GluNEO Lite, Onetouch select, and Vivachek fad. The results showed that
glucose concentrations in the control materials at three concentration levels (low, medium and high) in NSS and
saline-citrate buffer were not significantly different (p >0.05) by using automatic chemistry analyzer and four finger-
stick blood glucose meters, excepting GluNEO Lite. In addition, the control materials in NSS are better than saline-
citrate buffer by determination with finger-stick blood glucose meters. This study indicated that the matrices of
glucose control material effect glucose determination by using few finger-stick blood glucose meters.
Keyword: Finger-stick blood glucose meter, Glucose control material, Saline-sodium citrate buffer

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Page 96

Effects of boiling and storage on antioxidant activity and total phenolic content of ginger
juice

Kancharat Wanichnaranon, Wassachol Manusjun, Supissara Kongpech, Nontawit Pirat
Huachiew Chalermprakiet University

This study measured the antioxidant activity and total phenolic compounds of ginger.
Trolox Equivalent Antioxidant Capacity (TEAC) method, and Total Phenolic Content (TPC)
method using boiling under different conditions. Different brewing temperatures were studied
at 70, 80, 90 and 100 °C by TEAC method ranged 2,619.05-22,000 and the TPC method is in
the range 1,497-4,441.1 GAE/g of ginger the study found that boiling temperature had no
significant effect on TPC.
Keyword: DPPH, Trolox Equivalent Antioxidant Capacity, Total phenolic content

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Page 97

Protective role of natural products against oxidative stress in Caenorhabditis elegans Part 1

Pattapon Kittivithitkun1, Pornlada Chaijaroen1, Anchalee Prasansuklab2, Tewin Tencomnao3
1Program of Medical Technology, Faculty of Allied Health Sciences, Chulalongkorn University
2College of Public Health Sciences, Chulalongkorn University
3Department of Clinical Chemistry, Faculty of Allied Health Sciences, Chulalongkorn University

In modern days, medical technologies are massively developed, which results in an increase of the
life expectancy of people in the society. However, as people live longer, their body tends to progressively
degenerate with time. Although our understanding on aging is still limited, recent studies illustrate that aging is
associated with the accumulation of oxidative damage caused by an imbalance between the production of free
radicals and the capacity of antioxidant defense system. Hence, targeting oxidative stress using natural
products could be useful in treatment of aging and age-related pathologies.

Beijing grass (Murdannia loriformis) has been an important medicinal herb in traditional medicine for
centuries. It helps to support the immune system, detoxify the body, and aid in respiratory healing. Many studies
have revealed its effectiveness as an antioxidant agent. This plant is rich in flavonoids, which could prevent
free radicals and inhibit cancer growth. Levan, a polysaccharide active component of Erwinia herbicola, also
exhibited potential antioxidant properties. Nowadays, the nematode Caenorhabditis elegans has become one
of the leading model organisms for aging research. Nevertheless, both natural products have not been studied
for their protective roles in this model. Thus, we aimed to examine whether Beijing grass extracts or levan could
enhance oxidative stress resistance in C. elegans. In this study, Beijing grass was extracted by maceration with
hexane and ethanol, while levan was purchased. The wild-type N2 worms were incubated with extracts (12.5,
25, 50, 100 and 200 ug/ml) or levan (5, 10, 25 ug/ml) and monitored their survival under juglone-induced
oxidative stress. The number of live and dead worms were counted after 48 hours exposure and expressed as
percent viability. From our findings, it seems that none of the extracts or levan at any test concentrations showed
protective activity. Rather, the extracts appear to cause toxicity to the nematodes.
Keyword: Murdannia loriformis, Levan, Caenorhabditis elegans, Oxidative stress, Anti-aging

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Protective role of natural products against oxidative stress in Caenorhabditis elegans Part 2

Ariya Fujii1, Saowapark yahom1, Anchalee Prasansuklab2, Tewin Tencomnao3
1Program of Medical Technology, Faculty of Allied Health Sciences, Chulalongkorn University
2College of Public Health Sciences, Chulalongkorn University
3Department of Clinical Chemistry, Faculty of Allied Health Sciences, Chulalongkorn University

In modern days, medical technologies are massively developed, which results in an increase of the life
expectancy of people in the society However, as people live longer, their body tends to progressively degenerate
with time Although our understanding on aging is still limited, recent studies illustrate that aging is associated with
the accumulation of oxidative damage caused by an imbalance between the production of free radicals and the
capacity of antioxidant defense system Hence, targeting oxidative stress using natural products could be useful in
treatment of aging and age related pathologies

Beijing grass Murdannia loriformis has been an important medicinal herb in traditional medicine for
centuries It helps to support the immune system, detoxify the body, and aid in respiratory healing Many studies
have revealed its effectiveness as an antioxidant agent This plant is rich in flavonoids, which could prevent free
radicals and inhibit cancer growth Levan, a polysaccharide active component of Erwinia herbicola, also exhibited
potential antioxidant properties Nowadays, the nematode Caenorhabditis elegans has become one of the leading
model organisms for aging research Nevertheless, both natural products have not been studied for their protective
roles in this model Thus, we aimed to examine whether Beijing grass extracts or levan could enhance oxidative
stress resistance in C elegans In this study, Beijing grass was extracted by maceration with hexane and ethanol,
while levan was purchased The wild type N2 worms were incubated with extracts or levan in various concentrations
and monitored their survival under juglone induced oxidative stress The number of live and dead worms were
counted after 48 hours exposure and expressed as survival percentage viability Here we found that levan at the
concentrations of 50,100 and 200 µg ml tend to have the potential to defend against oxidative toxicity in nematodes,
whereas the extracts did not show any protective activity

Keyword Murdannia loriformis, Levan, Caenorhabditis elegans, Oxidative stress, Anti aging

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