Dian Eurike Septyaningtrias, Hilizza Awalina Zulfa, Mahayu Firsty, Ramadhani, Sumaryati, Dewi Sulistyawati, Dewi Kartikawati Paramita, Yustina Andwi Ari Sumiwi, Rina Susilowati


Gastrointestinal symptoms are common health problems found during aging and neurodegenerative diseases. Trimethyltin-induced rat is known as an animal model of hippocampal degeneration with no data on enteric neurodegeneration. This study aimed to investigate the effect of trimethyltin (TMT) induction on the gastrointestinal tract. A 28-day animal study with male Sprague–Dawley rats (3 months old, 150–200 g) given a single TMT injection (8 mg/kg body weight, intraperitoneal) was conducted. The number of neurons in the colonic myenteric plexus was measured using stereological estimation. Histological scoring of colon inflammation, immunohistochemistry of tumor necrosis factor-α (TNF-α), and quantitative PCR were conducted. This study showed neuronal loss in the colonic myenteric plexus of TMT-induced rat model of neurodegeneration. Minor colon inflammation characterized by inflammatory cell infiltration and slightly higher expression of TNF-α in the colon mucosa were observed in the TMT-induced rat. However, the gut microbiota composition of the TMT-induced rat was not different from that of the control rats. This study demonstrates that TMT induces colonic myenteric plexus neurodegeneration and minor colon inflammation, which suggests the potential of this animal model to elucidate the communication between the gastrointestinal tract and central nervous system in neurodegenerative diseases.

enteric nervous system, inflammation, myenteric plexus, neurodegeneration, rat colon, TNF-α, trimethyltin


Titis Nurmasitoh, Dwi Cahyani Ratna Sari, Rina Susilowati


Background: Moderate-intensity intermittent exercise (MIIE) has been proposed as an effective method for preventing Alzheimer’s dementia (AD). Aim: This study aimed to investigate the effects of MIIE on the spatial memory and protein level of AD markers in the hippocampus of trimethyltin (TMT)-induced rat model of hippocampal degeneration. Methods: Male Sprague Dawley (SD) rats were randomly assigned into four groups: normal control (N), exercise control (E), TMT control (T), and exercise and TMT (ET). Rats of the exercise groups (E and ET) were forced to run on a treadmill for 30 min each day at maximum for 12 weeks. Intraperitoneal injection of 8 mg/kgBW TMT was administered as a single dose, 10 days before the last exercise treatment for the T and ET groups. The spatial memory of rats was examined using Morris water maze (MWM) test after the exercise period. After euthanasia, the hippocampal tissue was dissected out and the level of hippocampal presenilin-1 (PSEN-1) and phosphorylated tau (p-tau) protein were measured using ELISA. The total number of hippocampal pyramidal neurons was estimated using unbiased stereological analysis. Qualitative immunohistochemistry was performed to examine the expression of brain-derived neurotrophic factor (BDNF), tumor necrosis factor-alpha (TNF-α), and interleukin-10 (IL-10) in paraffin sections of the hippocampus. Results: TMT exposure induced memory impairment indicated by the T group having the lowest percentage of time and percentage of path length in the target quadrant compared to other groups. MIIE prevented the memory impairment effect of TMT exposure indicated by the ET group having no significantly different MWM performance compared to the E and N groups. The ET group had significantly lower levels of hip
pocampal AD markers, p-tau and PSEN-1, as well as significantly higher estimated total number of pyramidal neurons of hippocampal CA1 and CA2–3 regions compared to the T group. Expressions of TNF-α was weak, while the expression of IL-10 was stronger in the ET group compared to the control group. The TMT-induced group exhibited stronger expression of BDNF.
Conclusion: MIIE prevents neuronal loss and impaired spatial memory upon TMT exposure most probably via preventing elevated levels of hippocampal AD markers and neuroinflammation.

KEYWORDS: Moderate-intensity intermittent exercise, Trimethyltin-induced neurodegeneration, Spatial memory, Aging hippocampus, Alzheimer’s disease markers


Gatot Soegiarto, Dewajani Purnomosari


In recent years, the elderly has become a rapidly growing proportion of the world’s population as life expectancy is extending. Immunosenescence and inflammaging contribute to the increased risk of chronic non-communicable and acute infectious diseases. Frailty is highly prevalent in the elderly and is associated with an impaired immune response, a higher propensity to infection, and a lower response to vaccines. Additionally, the presence of uncontrolled comorbid diseases in the elderly also contributes to sarcopenia and frailty. Vaccine-preventable diseases that threaten the elderly include influenza, pneumococcal infection, herpes zoster, and COVID-19, which contribute to significant disability-adjusted life years lost. Previous studies had shown that conventional vaccines only yielded suboptimal protection that wanes rapidly in a shorter time. This article reviews published papers on several vaccination strategies that were developed for the elderly to solve these problems: more immunogenic vaccine formulations using larger doses of antigen, stronger vaccine adjuvants, recombinant subunit or protein conjugated vaccines, newly developed mRNA vaccines, giving booster shots, and exploring alternative routes of administration. Included also are several publications on senolytic medications under investigation to boost the immune system and vaccine response in the elderly. With all those in regard, the currently recommended vaccines for the elderly are presented.
Keywords: elderly; frailty; immunosenescence; infection; recommended vaccine; comparative effectiveness analysis

Satrio Adi Wicaksono, Yustina Andwi Ari Sumiwi, Dewi Kartikawati Paramita, Rina Susilowati


Background: Studies in the digestive tract often required precision quantification of intestinal volume to observe the effect of certain intervention/
condition. Application of stereological methods could bring unbiased and accurate results but commercially computer-assisted systems are

not widely available. ImageJ-FIJI is an open source software, which could become an alternative choice in the stereological measurement

Aim: This study describes simple stereological quantification methods during volume estimation of jejunum-ileum intestinal layers
of the rats using a light microscope and ImageJ-FIJI stereological tool.
Material and Methods: Six 3-months old male Sprague-Dawley
rats were terminated and jejunum-ileum was harvested after perfusion. After removal of intestinal luminal content, whole jejunum-ileum

weight was measured. The organ was sampled as 6-10 slabs of 1 cm length in a systematic uniformed random sampling manner. Slabs were

cut longitudinally at random angles before flattened and put on filter papers for subsequent tissue processing into 2-3 paraffin blocks. One

section of 3 μm thick was sampled from each block, stained using toluidine blue and documented using a light microscope connected to a

microstepper apparatus. The volume of the intestinal layers was estimated using a point-counting grid on Image J-FIJI software.
Result: We
compared two sets of counting methods i.e. minimal counting (MC) and rigorous counting (RC) approaches that differ in their respective a/p

value. Quantification using RC approach resulted in significantly higher estimated volume of tunica submucosa and tunica muscularis while

having more preferable stereological accuracy parameters (CE<5% & CV<10%).
Conclusion: Although it required longer counting time,
rigorous approaches resulted in higher accuracy while still within the range of rule of thumb criteria of 0.2 < CE
2/CV2 < 0.5.

Keywords: Digestive tracts, ImageJFIJI, quantification, stereology, volume



Globally, the vaccine has been determined as one of the principal policies to tacklethe COVID-19 pandemic. However, some vaccinated individuals with two complete doses of inactivated experienced SARS-CoV2 infection, including the healthcare workers (HCWs). This threat led to the emergent need for a vaccine booster with different types ofplatforms aiming to enhance immunity from the Omicron variant. We conducted a literature study on the concept of heterologous compared to homologous vaccines in COVID-19 vaccination. We obtained 22 studies about COVID-19 booster vaccines. Referring to seven of them, we compared and distinguished between heterologous and homologous vaccines. We then reported the literature review according to PRISMA guideline. The study demonstrated qualitatively that heterologous vaccinations boosted antibody receptor binding domain, neutralizing antibody, and spike-specific Th1 type T cell responses and had an impact on omicron infection when compared to homologous vaccines. In conclusion,heterologous, mRNA based vaccine, predominantly induces cellular and humoral responses better than the homologous vaccine. This increased immune response is expected to provide profound immunity against the Omicron.

Keywords: vaccine, COVID-19, infectious disease, heterologous, boostervaccine, COVID-19, infectious disease, heterologous, booster

DOI: 10.20473/ijtid.v11i2.39597


Background: Diffuse large B-cell lymphoma (DLBCL) is the most common type of non-Hodgkin’s lymphoma. It is classified into homogeneous subtypes, namely Germinal Center B-Cell Like (GCB) and Non-Germinal Center B-Cell Like (GCB), with the latter associated with worse survival outcomes. Increased expression of Hypoxia-Inducible Factor-2α (HIF-2α) is often observed in DLBCL and has been correlated with enhanced angiogenesis, suggesting its potential as a prognostic factor in managing DLBCL. The study evaluates the association between HIF-2α overexpression and the GCB and non-GCB subtypes of DLBCL.

Methods: This cross-sectional study utilized samples from DLBCL patients at Dr. Kariadi Semarang Hospital between January and December 2021. Immunohistochemistry was performed on the samples to determine the DLBCL subtypes and assess the presence of HIF-2α. HIF-2α was evaluated by immunohistochemistry based on its distribution and intensity. The Kruskal-Wallis test was used to find the association between HIF-2α expression and the GCB and non-GCB subtypes of DLBCL. Furthermore, Spearman’s rank correlation test explored the correlation between all numeric variables.

Results: A total of 30 subjects were included in this study, with 7 samples of GCB subtype (23.3%) and 23 samples of non-GCB subtype (76.6%). Kruskal-Wallis test showed no association between HIF-2α expression and the subtypes of DLBCL (p=0.812). Spearman’s rho correlation test indicated no correlation between HIF-2α overexpression score with NCCN IPI score in GCB (r=0.219; p=0.637) and non-GCB (r=-0.194; p=0.386).

Conclusion: There was no association between HIF-2α expression and GCB and non-GCB subtypes of DLBCL. Additionally, no correlation was found between HIF-2α overexpression and NCCN IPI score.

KEYWORDS: DLBCL, HIP-2, GCB, non-CB, Lymphoma



Background: Microsatellite instability (MSI) is one of the important pathways involved in development of colorec-tal cancer (CRC). MSI occurs due to mutations or hypermethylation of negative MMR proteins MLH1 and PMS2. The accumulation of mutations at microsatellite locus accelerated the development of CRC. Characteristics of CRC patients with MSI are not sensitive to 5FU chemotherapy and have a good clinical outcomes that can be used as a prognostic factor and a predictor of therapy. Therefore MSI detection is needed.
Method: A retrospective cross-sectional study of 80 CRC slides obtained from DR. Sardjito Hospital Yogyakarta and clinical laboratory in 2010-2016. Immunohistochemical staining with anti-MLH1 and anti PMS2 antibodies to see MSI status. Negative MLH1 expression is called MSI MLH1 positive and PMS2 negative expression is called positive MSI PMS2. The association between MSI MLH1 and PMS2 with clinicopathology parameters was analyzed using Chi square.
Results: Colorectal cancer patients MSI MLH1 as much as 44 (61.1%), MSI PMS2 as much as 21 (29.2%) and, MSI MLH1 and PMS2 simultaneously as much as 18 (25.0%). MSI MLH1 and PMS2 positive are found in ≥50 years old. The number of male patients is more than women. Most of the patients had a T3-T4 tumor size with advanced stage and well differentiated. MSI MLH1 associated with tumor differentiation (p = 0.011).
Conclusion: MSI MLH1 is associated with tumor differentiation (p = 0.011), but, no association with the other clin-icopathology parameters. MSI PMS2 is not associated to the overall clinicopathology parameters. MSI MLH1 and PMS2 are not simultaneously associated with all clinicopathology parameters (p> 0.05).


Microsatellite Instability, MLH1, PMS2, Colorectal Cancer.

DOI: –

Ninda Devita, Adika Zhulhi Arjana, Umi Solekhah Intansari*, Rina Susilowati


This study aims to investigate NK cell number and cytokines level in various degrees of severity in COVID-19 cases. A total of 63 COVID-19 patient aged >18 y were divided into mild-moderate and severe-critical groups. Patient characteristics and peripheral blood count were obtained from medical records. NK cells number, levels of IFN-γ, IL-10, and IL-12 in peripheral blood were examined by means of flow cytometry. The severe-critical group had leukocytosis, neutrophilia, lymphopenia, higher Neutrophil Lymphocyte Ratio, lower NK cell number and higher level of IL-10. In severe-critical group, those aged >60 years had higher IL-10. In both groups, patients with diabetes comorbidities had a higher number of NK cells (p<0.05). NK cell number and IL-10 in peripheral blood have potential as a predictor of severe COVID-19 patients. Keywords: COVID-19; severity; NK cells; cytokines DOI:

Anggoro Budi Hartopo, Dyah Wulan Anggrahini, Lucia Kris Dinarti, Anne-Kathrin Schäfer, Andreas Bergmann, Jajah Fachiroh, Salvatore Di Somma


The adrenomedullin level increases in pulmonary arterial hypertension (PAH, and correlates with a high mortality rate. Its active form, bioactive adrenomedullin (bio-ADM), has been recently developed and has significant prognostic applications in acute clinical settings. Aside from idiopathic/hereditary PAH (I/H-PAH), atrial septal defects-associated pulmonary artery hypertension (ASD-PAH) is still prevalent in developing countries and associated with increased mortality. This study aimed to investigate the mortality-wise prognostic value of the plasma bio-ADM level by comparing subjects with ASD-PAH and I/H-PAH with ASD patients without pulmonary hypertension (PH) as a control group. This was a retrospective, observational cohort study. The subjects were Indonesian adult patients who were recruited from the Congenital Heart Disease and Pulmonary Hypertension (COHARD-PH) registry and divided into three groups: (1) ASD without PH (control group), (2) ASD-PAH and (3) I/H-PAH. During right-heart catheterization at the time of diagnosis, a plasma sample was taken and assayed for bio-ADM using a chemiluminescence immunoassay. Follow-up was performed as a part of the COHARD-PH registry protocol in order to evaluate the mortality rate. Among the 120 subjects enrolled: 20 turned out to have ASD without PH, 85 had ASD-PAH and 15 had I/H-PAH. Compared to the control group (5.15 (3.0–7.95 pg/mL)) and ASD-PAH group (7.30 (4.10–13.50 pg/mL)), bio-ADM levels were significantly higher in the I/H-PAH group (median (interquartile range (IQR)): 15.50 (7.50–24.10 pg/mL)). Moreover, plasma bio-ADM levels were significantly higher in subjects who died (n = 21, 17.5%) compared to those who survived (median (IQR): 11.70 (7.20–16.40 pg/mL) vs. 6.90 (4.10–10.20 pg/mL), p = 0.031). There was a tendency toward higher bio-ADM levels in those who died among the PAH subjects, in both ASD-PAH and I/H-PAH groups. In conclusion, the plasma bio-ADM level is elevated in subjects with PAH from both ASD-PAH and I/H-PAH origins, reaching the highest levels in subjects with the I/H-PAH form. A high bio-ADM level tended to be associated with a high mortality rate in all subjects with PAH, indicating a relevant prognostic value for this biomarker. In patients with I/H-PAH, monitoring bio-ADM could represent a valid tool for predicting outcomes, allowing more appropriate therapeutical choices.
Keywords:pulmonary arterial hypertension; idiopathic/heritable pulmonary arterial hypertension; atrial septal defects-associated pulmonary arterial hypertension; bioactive adrenomedullin; prognosis; mortality

Y Adani, J Fachiroh, FS T Dewi, MP Inggriani, AB Hartopo


Background and aim: The combination of environmental and genetic factors accounts for the developing of hypertension. Excessive salt intake and accumulation in the kidney can increase plasma volume and lead to high blood pressure. Sodium is consumed in various ways, such as cooking, processing and seasoning, This study was designed to verify the relationship between salt usage behaviour and blood pressure.
This cross-sectional study data were collected from the Health and Demographic Surveillance System in Sleman Regency from 2021 – 2022. The blood pressure were measured three times by an automatic blood pressure measuring device and the average values were recorded. Salt usage behaviors were surveyed by salt preference questionnaire. Relationships between salt consumption and hypertension were analyzed in SPSS ver. 26 for data analysis.
Among 1061 enrolled subjects (481 male and 580 female) with an average age of 59.19 + – 9.15 years old (p = 0.000). The highest systolic blood pressure was 221 mmHg and mean systolic blood pressure 140.20 + – 23.28 mmHg. Increase heart rate, body mass index, waist circumference, waist-to-hip ratio in hypertensive subjects were shown significant statistically (p = 0.000). The correlation between hypertension and salt usage behavior did not significantly differ between patients who used soy sauce before meal, additional of seasoning salt when cooking at home, consume salty snacks, and salt/ salty sauce that has been consumed with p = 0.039, p = 0.98, p = 0.82, p = 0.30, respectively.
These findings suggest that high salt intake does not independently associated with increased blood pressure in this population. Additional larger-scale and objective measurement of salt intake are needed for further clarification.

Keywords: high-salt intake, hypertension, salt usage behavior

DOI: 10.1097/01.hjh.0000935384.83137.65

Hilmi Muhammad, Sofia Mubarika Haryana, Rahadyan Magetsari, Aryadi Kurniawan, Bima Baikuni, Paramita Ayu Saraswati


Backgroud: Congenital Talipes Equinovarus (CTEV) is a multitude of deformities involving equinus, varus, adductus, and cavus deformities. Clubfoot affects 1 in every 1000 infants born worldwide, with various incidences according to geographical areas. It has been previously hypothesized that the possible genetic role in Idiopathic CTEV (ICTEV) might have a treatment-resistant phenotype. However, the genetic involvement in recurrent ICTEV cases is yet to be determined.

Aim: To systematically review existing literature regarding the discovery of genetic involvement in recurrent ICTEV to date to further understand the etiology of relapse.

Methods: A comprehensive search was performed on medical databases, and the review was conducted according to the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) 2020 guidelines. A comprehensive search was performed on several medical databases: PubMed (MEDLINE), Scopus, the Cochrane Library, and European PMC on May 10, 2022. We included studies reporting patients with recurring idiopathic CTEV or CTEV of unknown cause after treatment, reporting whole-genetic sequencing, whole-exome sequencing, Polymerase Chain Reaction, or Western blot analysis as methods of genetic analysis (inter-vention) and providing results of idiopathic CTEV genetic involvement. Non-English studies, literature reviews, and irrelevant articles were excluded. Quality and risk of bias assessments were performed using Newcastle-Ottawa Quality Assessment Scale for non- randomized studies where appropriate. The authors discussed data extracted with the primary outcome of gene(s) frequency being reported of their involvement in recurrent ICTEV cases.

Results: Three pieces of literature were included in this review. Two studies analyzed the genetic involvement in CTEV occurrence, while one analyzed the protein types found.

Discussion: With included studies of less than five, we could not perform other forms of analysis apart from qualitatively.Conclusion: The rarity of literature exploring the genetic etiology of recurrent ICTEV cases has been reflected in this systematic review, giving opportunities for future research.

Keywords: recurrent, idiopathic CTEV, idiopathic clubfoot, genetics, clubfoot


Gatot Soegiarto, Dewajani Purnomosari, Laksmi Wulandari, Bagus Aulia Mahdi, Karin Dhia Fahmita, et. al.


Background: The incidence of the Coronavirus Disease 2019 (COVID-19) among healthcare workers (HCWs) is widespread. It is important to understand COVID-19 characteristics among HCWs before and after vaccination. We evaluated the incidence of COVID-19 among HCWs in East Java, Indonesia comparing the characteristics of the disease between the pre- vs post-vaccination periods.

Methods: A retrospective observational study was conducted among HCWs in two major hospitals in East Java,
Indonesia, between April 01, 2020, and Oct 31, 2021. All HCWs were offered vaccination with inactivated viral vaccine (CoronaVac) from Jan 15, 2021. Therefore, we divided the time of the study into the pre-vaccination period (between April 01, 2020, and Jan 14, 2021) and post-vaccination period (between Jan 15 and Oct 31, 2021). We then compared the pattern of COVID-19 infections, and hospitalisations between these periods.

Findings: A total of 434 (15.1%) and 649 (22.6%) SARS-CoV-2 infections were reported among study participants (n = 2878) during the pre-vaccination and post-vaccination periods, respectively. The vaccine effectiveness was 73.3% during the first 3–4 months after vaccination but this decreased to 17.6% at 6–7 months after vaccination, which coincided with the emergence of the delta variant. The overall hospitalisation rate was reduced from 23.5% in the pre-vaccination period to 14.3% in the post-vaccination period. Hypertension appeared to be the strongest risk factor affecting hospitalisation in the pre-vaccination period. However, the risk due to hypertension was reduced in the post-vaccination period.

Interpretation: The risk to contract COVID-19 remains high among HCWs in East Java, Indonesia. Vaccination is important to reduce infection and hospitalisation. It is essentially important to evaluate the characteristics of COVID-19 infection, hospitalisation, the impact of co-morbidities and vaccine effectiveness in order to improve
the measures applied in protecting HCWs during the pandemic.

KEYWORDS: Healthcare workers; Vaccine; Hospitalisation; Co-morbidity


Anggoro Budi Hartopo, Maria Patricia Inggriani, Brilliant Winona Jhundy, Jajah Fachiroh, Putri Tiara Rosha, Ratri Kusuma Wardani, Fatwa Sari Tetra Dewi



There is a lack of data on modifiable coronary artery disease (CAD) risk factors in the Indonesian population, hindering the implementation of assessments and prevention programs in this population. This study investigated modifiable risk factors for CAD among Indonesians by comparing them between CAD-proven patients and healthy subjects from a similar population.
Methods : In this nested, matched case-control study, the cases were patients from a referral hospital in Yogyakarta, Indonesia and the controls were respondents in a population surveillance system in Yogyakarta, Indonesia. The cases were 421 patients who had undergone coronary angiography, showing significant CAD. The sex- and age-matched controls were 842 respondents from the Universitas Gadjah Mada Health and Health and Demographic Surveillance System Sleman who indicated no CAD presence on a questionnaire. The modifiable CAD risk factors compared between cases and controls were diabetes mellitus, hypertension, central obesity, smoking history, physical inactivity, and less fruit and vegetable intake. A multivariate regression model was applied to determine independent modifiable risk factors for CAD, expressed as adjusted odds ratios (AORs).
Results :
A multivariate analysis model of 1,263 subjects including all modifiable risk factors indicated that diabetes mellitus (AOR, 3.32; 95% confidence interval [CI], 2.09–5.28), hypertension (AOR, 2.52; 95% CI, 1.76–3.60), former smoking (AOR, 4.18; 95% CI, 2.73–6.39), physical inactivity (AOR, 15.91; 95% CI, 10.13–24.99), and less fruit and vegetable intake (AOR, 5.42; 95% CI, 2.84–10.34) independently and significantly emerged as risk factors for CAD.
Conclusions :Hypertension, diabetes mellitus, former smoking, physical inactivity, and less fruit and vegetable intake were independent and significant modifiable risk factors for CAD in the Indonesian population.
KEYWORDS: Coronary artery disease, Heart disease risk factors, Life style, Sedentary behavior, Dietary fiber


Nurvita Risdiana, Rina Susilowati, Eti Nurwening Sholikhah, Ginus Partadiredja


Erythrina subumbrans (Hassk.) Merr. is an alkaloid plant with dihydro-β-erythroidine (DhβE) content which is considered to block α4β2 nAChRs subtype and, therefore, may suppress the desire to use nicotine. This study aimed to investigate these possible effects of E.subumbrans (Hassk.) Merr. extract on nicotine withdrawal syndrome and β2 nAChRs expression in rats’ ventral tegmental area (VTA). The rats were divided into six groups, i.e., control (OO), nicotine treated (NO), nicotine, and E. subumbrans (Hassk.) Merr.-treated (NE 100, NE 200, NE 400), and E. subumbrans (Hassk.) Merr.treated (OE 200) groups. Nicotine was given ad libitum via drinking water with a step-wise increase of dosage every four days for 30 days. Somatic and affective signs were observed during the dark cycle of 24 hours abstinent period (days 31and 46). The expression of β2 nAChRs in the VTA was examined semi-quantitatively. It has been found that the rearing behavior of the NE 100 group was fewer on day 46 than on day 31. The body scratching behavior of the NE 100 group was fewer than that of the OO group on day 46. The front paws and penile licking behaviors of the NE 100 group were fewer than those of the NO group on day 46. The open arm entries of the NO group were fewer than that of the NE 200 group on day 46. The β2nAChRs expression of the NO group was lower than that of the OO group. E. Subumbrans (Hassk.) Merr. at a dosage of 100mg/kg BW may decrease some somatic signs of nicotine withdrawal syndrome.



Tirta Wardana, Siti Nur Chasanah, Risky Oktriani, Cita Herawati, Sumadi Lukman Anwar, Indwiani Astuti, Sofia Mubarika Haryana



Background: Nasopharyngeal carcinoma (NPC) is endemic cancer in Southeast Asia with a relatively poor
prognosis. Chemoradiotherapy is a primary treatment that advantages certain patients, particularly in the early
stages. New predictive and prognostic biomarkers are required to guide and select the best treatment.
Aims: To evaluate the circulation expression profile of microRNAs (miRNAs) associated with responses to che-
moradiotherapy in nasopharyngeal carcinoma.
Methods: Peripheral blood from 17 patients was collected before and after chemotherapy and radiotherapy.
Differential expression circulating miRNAs were analyzed using microRNA Cancer Panels and were compared
among patients with complete responses. Differential expression analysis using GenEx 7 Multid, statistic rep-
resented by GraphPad Prism 9. Alterations mechanism signaling pathways and biological function using IPA
(Ingenuity Pathways Analysis).
Results: Using microRNAs Cancer Plate consisting of 116 miRNAs, we identified ten circulating miRNAs that were
differentially expressed in NPC patients after chemoradiotherapy. Unsupervised clustering and confirmation
using qRT-PCR showed that miR-483-5p, miR-584-5p, miR-122-5p, miR-7-5p, miR-150-5p were overexpressed
and miRNA are miR-421, miR-133a-3p, miR-18a-5p, miR-106b-3p, miR-339-5p were significantly down-
regulated after chemoradiotherapy (p < 0.0001). In addition, ROC analysis through AUC (Area Under Curve) with 99% confidence interval (CI) p value < 0.0001. Gene enrichment analysis of microRNAs and the targeted proteins revealed that the main involved pathways for chemoradiotherapy in NPC were cell death and survival signaling pathways. Conclusion: qPCR profiling in circulating blood compared before and after chemoradiotherapy in nasopharyngeal carcinoma can identify pathways involved in treatment responses. miR-483-5p, miR-584-5p, miR-122-5p, miR-7- 5p, miR-150-5p, miR-421, miR-133a-3p, miR-18a-5p, miR-106b-3p, miR-339-5p are differentially regulated after chemoradiotherapy in NPC. KEYWORDS: DOI:

Ryo Shirakashi, Zisis Kozlakidis, Birendra Kumar Yadav, Wayne Ng, Jajah Fachiroh, Hanh Vu, Tatsuaki Tsuruyama, Koh Furuta



Calls to reduce or entirely remove the carbon footprint of ongoing activities, collectively termed as decarbonization, have become increasingly more vocal in health care with a number of recent, high profile consensus statements. These calls encourage the biobanking field, as one of the foundational health care research infrastructures, to consider decarbonization as a potential novel research area both in terms of the molecules and the equipment used in research. The current article provides a summary of the roundtable discussion during the 2022 ISBER Annual Meeting and Exhibits, highlighting the current knowledge gaps, challenges, and opportunities in this field. In particular, technological innovation, a greater awareness of the current situation, and behavioral change are important pieces of the puzzle to improving the future of decarbonization in biobanking, even if the eventually implemented routes between resource-abundant and resource-restricted settings might be distinctly different. This article sets the foundation for raising awareness of the subject and of subsequent steps that need to be undertaken.



Maria Patricia Inggriani, Ahmad Musthafa, Ira Puspitawati, Jajah Fachiroh, Fatwa Sari Tetra Dewi, Anggoro Budi Hartopo



Diabetes mellitus (DM) increases risk of coronary artery disease (CAD). Endothelin-1 (ET-1) is a potential biomarker of endothelial dysfunction. This study aimed to evaluate ET-1 level in CAD patients and its relationship with DM. The cross-sectional design included subjects with angiographically proven CAD and controls among Indonesian. DM was defined by medical history and anti-diabetics use. Serum ET-1 level was measured in both subject groups. We recruited 305 subjects, 183 CAD patients and 122 controls. CAD subjects had higher percentage of males, DM, hypertension, dyslipidemia, smoking, family history of cardiovascular disease, and obesity. ET-1 level was significantly higher in CAD than in controls (2.44 ± 1.49 pg/mL vs. 1.76 ± 0.83 pg/mL; p < 0.001). Increased ET-1 level was significantly associated with DM and dyslipidemia. The highest ET-1 level was observed in CAD with DM, followed by CAD non-DM (2.79 ± 1.63 pg/mL vs. 2.29 ± 1.40 pg/mL; p = 0.023). Among controls, ET-1 level was the lowest in non-DM subjects. Female CAD had higher proportion of DM; however, ET-1 level was similar to male CAD with DM. In conclusion, an increased ET-1 level was significantly associated with DM in patients with CAD. Further research should investigate the potential role of ET-1 receptor antagonists in the secondary prevention of CAD with DM.



Fery L Widiany, Marsetyawan Soesatyo, Lily Arsanti Lestari, Woro Rukmi Pratiwi, Mae Sri Hartati Wahyuningsih, Emy Huriyati


Background: Hemodialysis patients can experience problems, including proteinenergy malnutrition, infection, disorders of the immune system, and inflammation. One etiology of malnutrition in hemodialysis is inadequate energy and protein intake, making patients need nutritional support, which can be fulfilled by local Indonesian foodstuffs.

Objective: This paper aimed to review the potential health benefits of snails (Pila ampullacea), tempeh, and Moringa oleifera leaves as nutritional support for hemodialysis patients.

Methods: In this review, the methodology used was based on comprehensive data searched from PubMed for literature review and technology benchmarking in making nutritional support for hemodialysis patients. An in-depth discussion, including the advantages and drawbacks of each foodstuff, is presented and outlined. Furthermore, key solutions are proposed and presented to overcome hemodialysis issues.

Results: The mixture of snail, tempeh, and moringa leaves as nutritional support for hemodialysis patients shows a good combination of natural ingredients from animals and plants. The nutritional content of the three mixed ingredients is found to meet the dietary requirements of hemodialysis, which are high protein, calcium and antioxidants, low phosphorus, and a phosphorus-toprotein ratio of <16. Conclusion: The combination of snails, tempeh, and moringa leaves provides several potential health benefits in overcoming nutritional problems, decreased immune status, and inflammation of hemodialysis patients. DOI:

Suci Widhiati, Shinta Trilaksmi Dewi, Retno Danarti, Hardyanto Soebono, Yulia Eka Irmawati, Monika Puspitasari, Niken Trisnowati, Tri Wibawa, Dewajani Purnomosari, Yohanes Widodo Wirohadidjojo


Background: Autologous non-cultured cell (ANCC) spray has been used to treat burns, chronic wounds, and vitiligo, but its use in junctional epidermolysis bullosa (JEB) has not been published previously. Chronic wounds in JEB are caused by mutations of laminin 332 (L322), whose function is to attach and act as a glue in the basal membrane. It is proposed that ANCC applications can provide keratinocytes and fibroblasts required to improve epithelization and spontaneously correct revertant keratinocytes in the wound area.
Purpose: To develop a modified procedure of ANCC spray and improve epithelization using silver sulfadiazine covered with plastic wrap to treat chronic wounds of JEB.
Patients and Methods: Shave excision of the donor site was performed on a 19-year-old girl with JEB. The ANCC spray was prepared and applied to the chronic wound, which was then covered with silver sulfadiazine occluded with plastic wrap.
Results: Following the ANCC spray application, epithelization was successfully initiated. Unfortunately, the wounds recurred after four months of follow-up.
Conclusion: The modified application method of ANCC spray provides a good alternative to treat chronic wounds in JEB.
Keywords: revertant mosaicism, silver-sulfadiazine, plastic wrap, chronic wound, junctional epidermolysis bullosa


Eggi Arguni, Fatwa Sari Tetra Dewi, Jajah Fachiroh, Dewi Kartikawati Paramita, Septi Kurnia Lestari, Bayu Satria Wiratama, Annisa Ryan Susilaningrum, Bara Kharisma, Yogi Hasna Meisyarah, Merlinda Permata Sari, Zakiya Ammalia Farahdilla, Siswanto Siswanto, Muhammad Farhan Sjaugi, Teguh Haryo Sasongko, Lutfan Lazuardi


The long-term antibody response to the novel SARS-CoV-2 in infected patients and their residential neighborhood remains unknown in Indonesia. This information will provide insights into the antibody kinetics over a relatively long period as well as transmission risk factors in the community. We aim to prospectively observe and determine the kinetics of the anti-SARS-CoV-2 antibody for 2 years after infection in relation to disease severity and to determine the risk and protective factors of SARS CoV-2 infections in the community. A cohort of RT-PCR confirmed SARS-CoV-2 patients (case) will be prospectively followed for 2 years and will be compared to a control population. The control group comprises SARS-CoV-2 non-infected people who live within a one-kilometer radius from the corresponding case (location matching). This study will recruit at least 165 patients and 495 controls. Demographics, community variables, behavioral characteristics, and relevant clinical data will be collected. Serum samples taken at various time points will be tested for IgM anti-Spike protein of SARS-CoV-2 and IgG anti-Spike RBD of SARS-CoV-2 by using Chemiluminescent Microparticle Immunoassay (CMIA) method. The Kaplan-Meier method will be used to calculate cumulative seroconversion rates, and their association with disease severity will be estimated by logistic regression. The risk and protective factors associated with the SARS-CoV-2 infection will be determined using conditional (matched) logistic regression and presented as an odds ratio and 95% confidence interval.


Tirta Wardana, Risky Oktriani, Cita Herawati Murjayanto, Denise Utami Putri, Sumadi Lukman Anwar, Teguh Aryandono, Sofia Mubarika Haryana


Background and aim: Nasopharyngeal Carcinoma (NPC) is an upper respiratory tract cancer prevalent in Southeast Asia and related to chronic EBV infection. microRNAs (miRNAs) regulate gene expression implicated in NPC’s carcinogenesis . However, this circulating RNA molecule’s role and clinical utility remain unknown. Therefore, this study examined the circulation of miRNAs and their association with clinical data.

Methods: 160 plasma samples of NPC and 80 non-tumor samples were extracted to evaluate and validate the gene expressions. Quantification expression was performed using relative quantification of qPCR analysis level expression methods. The intrinsic cellular roles involving biological signaling in NPC’s oncogenesis using Ingenuity Pathways Analysis (IPA) were also used.

Results: The results of the quantification significance profiling of NPC samples revealed decreased miR-29c-3p (fold change 1.16; p<0.05) and increased 195-5p expression (fold change 1.157; p<0.05). Furthermore, the validation of hsa-miR-29c-3p expression on plasma NPC with known tumor vs non-tumor and significant changes was also performed using a fold change of 4.45 (medians of 31.45±1.868 and 24.96±1.872, respectively; p<0.0005). miR-29c had a 2.14 fold change correlated with T primary status with a median of 31.99±1.319 and 31.35±2.412, respectively (p<0.05). Stage status with fold change 1.99 also had median levels of 31.98±1.105 and 31.21±2.355, respectively (p-value <0.05). Furthermore, the node's status for the lower expression of miR-29c with fold change 1.17 had median levels of 32.78±2.221 and 31.33±1.689, respectively (p-value of 0.7). Bioinformatics analysis established the roles and functions of miR-29 in NPC progression, cell death and survival, cellular development, cellular function, and cell maintenance by inhibiting COL4A, PI3K, VEGFA, JUN, and CDK6. Conclusion: Overall, we conclude that decreased miR-29c expression is associated with poor clinical status and might inhibit NPC’s five target genes.

Keywords: MicroRNA; cancer; circulating.; clinical outcome; nasopharyngeal; profiling.


Awal Prasetyo, Udadi Sadhana, Dewi K Paramita, Sofia Mubarika Haryana, Bambang Hariwiyanto, Soenarto Sastrowijoto, Totok Utoro


BACKGROUND: The risk-combination of genetic or familial history, environmental risk factors, and EBV infection might cause nasopharyngeal carcinogenesis. The serum antibody for EBV IgA, namely, EBNA1+VCA-p18 has a good sensitivity as an early diagnostic test for nasopharyngeal carcinoma (NPC).

AIM: This study aims to determine the correlation between risk factors and histopathological typing of NPC and also the correlation between the IgA [EBNA-1 + VCA p-18] ELISA and histologic type.

METHODS: A cross-sectional method was used on 108 NPC patients which filled a questionnaire through an in-depth interview on the family condition to cancer, habit/lifestyle, and environmental risks. A total of 47 subjects were willing to donate blood samples for IgA [EBNA1 + VCA p-18] ELISA. Furthermore, Kendall’s tau-b (τ) correlation test was performed on NPC keratin type (WHO-1) and non-keratin (WHO-2 and 3).

RESULTS: The results showed that the family history of non-keratinized NPC was associated with NPC WHO-3 as demonstrated by τ = 0.473, as well as salt-eating with τ = 0.334, smoked/grilled fish/meat eating τ = 0.205, instant noodle-eating τ = 0.356, consuming canned/packaged canned foods τ = 0.240, and flavored food eating habits τ = 0.364, along with passive smoking τ = 0.377, and chronic nasopharyngeal infection τ = 0.530. The IgA titers, namely, [EBNA1 + VCA p-18] ELISA for non-keratin type NPC was greater than the keratin type; however, it was not related to WHO-3 NPC as indicated by τ = 0.376, and p = 0.011 put this underlying before however.

CONCLUSIONS: The positivity of IgA [EBNA-1 + VCA p-18] ELISA does not correlate with the non-keratin type histologic NPC, family history, as well as salt-eating, instant noodle, and flavored food eating habits, along with passive smoking and nasopharyngeal infection.


Dwita Anastasia Deo, Elizabeth Henny Herningtyas, Umi Solekhah Intansari, Taufik Mulya Perdana, Elsa Herdiana Murhandarwati, Marsetyawan HNE Soesatyo


Microscopic examination is the backbone of malaria diagnosis and treatment evaluation in Indonesia. This test has limited ability to detect malaria at low parasite density. Inversely, nested polymerase chain reaction (PCR) can detect parasites at a density below the microscopic examination’s detection limit. The objective of this study is to compare microscopic and PCR results when being used to identify malaria in suspected patients and patients who underwent dihydroartemisinin–piperaquine (DHP) therapy in the last 3–8 weeks with or without symptoms in Sumba Barat Daya, Nusa Tenggara Timur, Indonesia. Recruitment was conducted between April 2019 and February 2020. Blood samples were then taken for microscopic and PCR examinations. Participants (n = 409) were divided into three groups: suspected malaria (42.5%), post-DHP therapy with fever (4.9%), and post-DHP therapy without fever (52.6%). Microscopic examination found five cases of P. falciparum + P. vivax infection, while PCR found 346 cases. All microscopic examinations turned negative in the post-DHP-therapy group. Conversely, PCR result from the same group yielded 29 negative results. Overall, our study showed that microscopic examination and PCR generated different results in detecting Plasmodium species, especially in patients with mixed infection and in patients who recently underwent DHP therapy.

Keywords: subclinical malaria, asymptomatic malaria, high endemicity, nested PCR, microscopic examination

Birendra Kumar Yadav, Wayne Ng, Hanh Vu, Jajah Fachiroh, Tatsuaki Tsuruyama, Li Zhou, Marianne K Henderson, Sharvari Gokhale, Koh Furuta


Biobanking is a relatively newly recognized and innovative branch of science, which includes the collection of samples and associated data from hospitals, diagnostic centers, and voluntary donations for biomedical and environmental research. It involves diverse stakeholders at the junction of society, science, ethics, law, and politics. A key element in the success of a biobank is the trust and support of public donors, clinicians, and scientists. To achieve trust, it is important to implement strategies that can increase biobank awareness in common people, and different types of communities. Biobank laws and regulations and transparent governance by the biobanks are also crucial to achieving public trust.

Keywords: awareness; donors’ trust in a biobanking; transparency; trust.



Mutiara Putri, Anggoro Budi Hartopo, Maria Patricia Inggriani, Jajah Fachiroh, Fatwa Sari Tetra Dewi


Background: Hypertension is known as independent factor in correlation with coronary artery disease (CAD) and play important part in atherosclerotic process. In an animal model with hypertension which endothelin-1 play as a vasoconstrictor, there was overexpression of endothelin-1 in the vessel walls. This overexpression suggests a role of endothelin-1 in hypertension patient, especially among CAD.

Material and methods: This was a cross-sectional study. A total 226 subjects were analysed, consisted of 127 subjects with CAD and 99 healthy population. The CAD subjects were patients underwent elective coronary angiography with significant CAD lesion. The healthy population were respondents of Sleman-HDSS survey (year 2019). Hypertensive subjects were defined those with history of hypertension from anamnesis. Diabetic subjects were excluded. The endothelin-1 was measured from peripheral serum samples by ELISA method. The comparative analysis was performed with Mann-Whitney test and the correlation was performed with Spearman correlation test.

Results: Mean serum endothelin-1 level was 2.1±1.2 pg/mL in hipertensive and 2.6±1.6 pg/mL in normotensive (p=0.063) among CAD subjects. Among healthy population, mean serum endothelin-1 level was 1.7±0.7 pg/mL in hypertensive and 1.8±0.8 pg/mL) in normotensive, (p=0.675). In addition, Spearman correlation between serum endothelin-1 and systolic blood pressure showed correlation coefficient —0.045 (p = 0.543) in CAD subjects and -0.165 (p=0.069) in healthy population which indicated inverse correlation between those parameters in both populations.

Conclusion: Serum endothelin-1 level did not differ significantly based on hypertensive status both in CAD and healthy population. There was a tendency toward decreased serum endothelin-1 level in hypertensive subjects.

DOI: 10.1097/01.hjh.0000832996.66949.02

Birendra Kumar Yadav, Hanh Vu, Jajah Fachiroh, Tatsuaki Tsuruyama, Wayne Ng, Koh Furuta


Statement of the Problem: Several standards and guidelines for biobanks or biorepositories have been published by various parties (e.g., the International Society for Biological and Environmental Repositore [ISBER] and the International Organization for Standardization [ISO]). These documents are invaluable for improving the routine practices of the biobanks but the implementation has proven to be challenging for those biobanks from the non-English regions because these resources are mostly written in English.

Proposed Solution: The World Health Organization (WHO) has recently published the International Classification of Diseases 11th Revision (ICD-11) along with a translation tool (lexique) for potential users. This has inspired us to make a similar contribution in the biobanking field. All the regional ambassadors (RAs) and director-at-large (DAL) in the Indo-Pacific Rim (IPR) region worked together to produce a similar lexique for potential users of ISBER’s Best Practices (BPs) 4th edition. A lexique with languages of Hindi, Indonesian, Vietnamese, and Japanese has been prepared.

Conclusions: This lexique is a comparison table between various languages and is expandable to other languages. In addition, this lexique will be a good tool for understanding the ISBER BPs 4th edition.

Keywords: ISO 20387; best practice; lexique; translation.


Astri Ferdiana, Jajah Fachiroh, Dyah Ayu Mira Oktarina, Astrid Irwanto, Caroline Mahendra, Sri Awalia Febriana, Hardyanto Soebono


Background: Allopurinol is the most commonly used drug for the treatment of gout arthritis. However, the use of allopurinol is associated with severe cutaneous adverse reactions (SCARs) and life-threatening immune-mediated reactions that include Stevens-Johnson syndrome (SJS). SJS induced by allopurinol is strongly linked with the presence of HLA-B*58:01 in the Asian population. Such a study has not been conducted in Indonesia. We present two cases with clinical diagnosis of SJS. These patients had Javanese ethnicity, for which evidence on the genetic predisposition of allopurinol-induced SJS/TEN had not been established. Testing for the presence of the HLA-B∗58:01 allele was positive in both cases. Our case report confirms findings from studies in Asian countries that link HLA-B*58:01 and allopurinol-induced SJS/TEN. A larger study is needed to elicit evidence that the HLA-B*58:01 allele can potentially be used as a genetic marker for allopurinol-induced SCARs among different ethnicities in Indonesia.

Keywords: HLA-B*58:01; adverse drug reaction; allopurinol; pharmacogenetics; severe cutaneous adverse reactions; stevens-johnson syndrome.


Anggi Laksmita Dewi, Dewi Kartikawati Paramita, Jajah Fachiroh


Rs1948 A>G is a single nucleotide variation (SNV) in the 3’‐UTR of CHRNB4. Genotyping the synonymous CHRNB4 rs1948 may be useful in identifying a lung cancer susceptibility gene. The study aimed to develop a simple and easy tetra‐primer amplification refractory mutation system (ARMS PCR) for CHRNB4 rs1948. The following steps were taken to optimize tetra‐primer ARMS PCR: 1) determining the gene sequence and position of a single mutation; 2) developing outer and inner primers; 3) amplification of target gene fragments via PCR using an outer primer; 4) genotyping PCR product using Sanger sequencing; 5) determining the optimal annealing temperature and PCR cycle; 6) determining optimal outer and inner primer ratio; and 7) testing the reproducibility of the PCR program and final validation with Sanger sequencing. Genotype (PCR result) was visualized with 3% agarose gel electrophoresis. Optimum condition was determined as annealing temperature of 64.8 ºC and 35 cycles, outer and inner primer ratio of 1:6, and DNA volume of 3 μL. Sanger sequencing confirmed the results of the tetra‐primer ARMS PCR and it was shown that ARMS PCR was able to identify three different variants of CHRNB4 rs1948.
KEYWORDS CHRNB4; genetic variant; genotyping; nicotinic receptor; tobacco smoking; rs1948


Sandy Nur Vania Putri, Aditya Rifqi Fauzi, Dewi Kartikawati Paramita, Ishandono Dachlan, Rosadi Seswandhana


Background Traumatic brain injury (TBI) is one of the leading causes of death and disability worldwide. In various combinations, a high frequency of facial bone fractures has been reported. However, the results are still conflicting. Methods We retrospectively reviewed medical records of patients with maxillofacial trauma who were admitted to Dr. Sardjito Hospital, Yogyakarta, Indonesia, between January 2016 and December 2017. Results A total of 70 patients with maxillofacial trauma were involved (57 males and 13 females, 18–65 years). Moreover, most of them were 18–25-year-old males (34.3%). The average patient age was 35.5 ± 14 years. No significant association was found between the sex and age group of the patient (p = 0.774). Motorcycle accident was the most frequent cause of maxillofacial fractures (84%) with midfacial and multiple maxillofacial fractures being the most frequent types found in patients (40% and 40%, respectively). The most common facial fracture was in the zygomatic bone (28%) and most cases showed mild facial injury (81%). A significant association was found between the Glasgow Coma Scale (GCS) and types of
traumatic brain injury (TBI) (p = 0.031). However, when GCS was compared with facial injury, no statistically significant level was reached and its correlation was low (p = 0.267, r = 0.134). Conclusions There was a significant association between types of traumatic brain injury with head injury severity. However, we found no correlation between head injury severity and facial injury severity. To explain and validate our results, further multicenter studies with a larger sample size are required.
Level of evidence: Level IV, Risk/Prognostic.
Keywords Maxillofacial injuries · Maxillary fractures · Craniocerebral trauma · Intracranial hemorrhages · Glasgow Coma Scale


Gatot Soegiarto, Laksmi Wulandari, Dewajani Purnomosari, Karin Dhia Fahmita, Hendra Ikhwan Gautama, Satrio Tri Hadmoko, Muhammad Edwin Prasetyo, Bagus Aulia Mahdi, Nur Arafah, Dewi Prasetyaningtyas, Pujo Prawiro Negoro, Cita Rosita Sigit Prakoeswa, Anang Endaryanto, Desak Gede Agung Suprabawati, Damayanti Tinduh, Eka Basuki Rachmad, Erwin Astha Triyono, Joni Wahyuhadi, Catur Budi Keswardiono, Feby Elyana Wardani, Fitriyah Mayorita, Nunuk Kristiani, Ari Baskoro, Deasy Fetarayani, Wita Kartika Nurani, Delvac Oceandy


Several types of vaccines have been developed to prevent the coronavirus disease 2019 (COVID-19). It is important to understand whether demographic and clinical variables affect the effectiveness of various types of vaccines. This study analysed the association between demographic/clinical factors, antibody response and vaccine effectiveness in healthcare workers vaccinated with inactivated virus. We enrolled 101 healthcare workers who received two doses of inactivated viral vaccine (CoronaVac). Blood samples were analysed at 1, 3, and 5 months after the second dose of vaccination. Data regarding demographic characteristics, medical histories, and clinical parameters were collected by interview and medical examination. In a separate retrospective study, we analysed the incidence of vaccine breakthrough infection on 2714 healthcare workers who received two doses of inactivated viral vaccine. Medical histories and demographic data were collected using a structured self-reported questionnaire. We found that antibody titres markedly increased at 1 month after vaccination but gradually decreased at 3-5 months post-vaccination. We observed a significant association between age (≥40 years) and antibody level, whereas sex and body mass index (BMI) exhibited no effect on antibody titres. Amongst clinical variables analysed, high blood pressure and history of hypertension were significantly correlated with lower antibody titres. Consistently, we found a significant association in the retrospective study between hypertension and the incidence of breakthrough infection. In conclusion, our results showed that hypertension is associated with lower antibody titres and breakthrough infection following COVID-19 vaccination. Thus, blood pressure control might be important to improve the efficacy of inactivated virus vaccine.

Keywords: Antibody response; Breakthrough infection; COVID-19; Comorbidity; Hypertension; Inactivated viral vaccine.


Chia-Wen Lin, Dian E Septyaningtrias, Hsu-Wen Chao, Mikiko Konda, Koji Atarashi, Kozue Takeshita, Kota Tamada, Jun Nomura, Yohei Sasagawa, Kaori Tanaka, Itoshi Nikaido, Kenya Honda, Thomas J McHugh, Toru Takumi


Objective: This study aimed to investigate the association between intra-tumoral and stromal VDR expressions with molecular subtypes and clinicopathological factors. Methods: A total of 75 formalin-fixed paraffin embedded tissue samples were stained using immunohistochemical methods. The VDR expressions were measured by counting brown-stained nuclei in intra-tumoral and stromal areas. The association of VDR expression with molecular subtypes and clinopathological factors was examined. Statistical analysis was performed by chi square tests.

High intra-tumoral VDR expression was found in carcinomas with luminal molecular subtypes (p=0.039) and low
histological degrees (p=0.035). High VDR expression in the stroma was found in breast carcinomas with large tumor sizes. Conclusions: High intra-tumoral VDR expression is found in breast carcinomas with luminal subtypes and low histological grade (I/II). Both factors are known to have a good prognosis. These findings further strengthen the function of VDR as anti-tumorigenesis.

Keywords: Breast carcinoma- tumor microenvironment- vitamin D receptor- clinicopathological factors

DOI: 10.31557/APJCP.2022.23.4.1169

Sukma Diani Putri, Siti Rahma Yunianda Nanza, Irianiwati Widodo, Dewajani Purnomosari


Objective: This study aimed to investigate the association between intra-tumoral and stromal VDR expressions with molecular subtypes and clinicopathological factors.

Methods: A total of 75 formalin-fixed paraffin embedded tissue samples were stained using immunohistochemical methods. The VDR expressions were measured by counting brown-stained nuclei in intra-tumoral and stromal areas. The association of VDR expression with molecular subtypes and clinopathological factors was examined. Statistical analysis was performed by chi square tests.

High intra-tumoral VDR expression was found in carcinomas with luminal molecular subtypes (p=0.039) and low
histological degrees (p=0.035). High VDR expression in the stroma was found in breast carcinomas with large tumor sizes. Conclusions: High intra-tumoral VDR expression is found in breast carcinomas with luminal subtypes and low histological grade (I/II). Both factors are known to have a good prognosis. These findings further strengthen the function of VDR as anti-tumorigenesis.

Breast carcinoma- tumor microenvironment- vitamin D receptor- clinicopathological factors

DOI: 10.31557/APJCP.2022.23.4.1169

Indah K. Murni, Dian C. Sulistyoningrum, Rina Susilowati, Madarina Julia, and Kacie M. Dickinson


Background and objective: Poor diets, characterized by excess fat, sugar and sodium intakes, are considered to be one of the most important modifiable risk factors for cardiovascular disease. Diet patterns and intakes during adoles-cence may persist into adulthood and impact on risk for chronic disease later in life. We aimed to evaluate the dietary intake of obese adolescents and its relationship to cardiometabolic health including lipid status and glycemic control. Methods and study design: This was a cross-sectional study of obese children aged 15 to < 18 years in Yogyakarta, Indonesia. All children had a medical history performed including a physical examination and fasting blood sample. Dietary intake was assessed using a semi-quantitative recall food frequency questionnaire. Multivariable linear regression model was performed to determine the relationship between dietary intakes and cardiovascular disease risks and to adjust for potential confounders. Results: Of 179 adolescents, 101 (57.4%) were male and median age was 16.4 (15.0–17.9) years. The majority of ado-lescents (98%) had inadequate intake of fibre and exceeded intakes of total fat (65%) and total sugar (36%). There was statistically significant correlation found in the multivariable linear regression analysis between fibre intake and HDL cholesterol after adjusting for potential confounders (β = 0.165; p = 0.033). Conclusions: This study demonstrates that there is a high proportion of obese Indonesian adolescents with poor dietary intakes. There was relationship observed between intake of nutrients of concern (fibre) and cardiometabolic risk factor among this sample of obese adolescents. Future research should examine overall dietary patterns in more detail among this population to elucidate the role of poor diet intakes in development of cardiovascular disease risk factors in young people transitioning into adulthood.
Keywords: Dietary intake, Cardiovascular disease, Obese, Adolescents, Indonesia


Eggi Arguni, Endah Supriyati, Mohamad Saifudin Hakim, Edwin Widyanto Daniwijaya, Firdian Makrufardi, Ayu Rahayu, Anwar Rovik, Utari Saraswati, Farida Nur Oktoviani, Nenes Prastiwi, Titik Nuryastuti, Tri Wibawa, Sofia Mubarika Haryana



Growing evidence shows that viral co-infection is found repeatedly in patients with Coronavirus Disease–2019 (COVID-19). This is the first report of SARS-CoV-2 co-infection with viral respiratory pathogens in Indonesia.


Over a one month period of April to May 2020, SARS-CoV-2 positive nasopharyngeal swabs in our COVID-19 referral laboratory in Yogyakarta, Indonesia, were tested for viral respiratory pathogens by real-time, reverse transcription polymerase chain reaction (RT-PCR). Proportion of co-infection reported in percentage.


Fifty-nine samples were positive for other viral respiratory pathogens among a total of 125 samples. Influenza A virus was detected in 32 samples, Influenza B in 16 samples, Human metapneumovirus in 1 sample, and adenovirus in 10 samples. We did not detect any co-infection with respiratory syncytial virus. Nine (7.2%) patients had co-infection with more than two viruses.


Viral co-infection with SARS-CoV-2 is common. These results will provide a helpful reference for diagnosis and clinical treatment of patients with COVID-19.

Key Words: COVID-19, SARS-CoV-2, Co-infection, Viral respiratory pathogens, Coronavirus


Denise U Putri, Po-Hao Feng, Chiou-Feng Lin, Sofia M Haryana, Marsetyawan HNE Soesatyo, Kang-Yun Lee, Chia-Li Han


Local administration of attenuated mycobacterium has been used as a cancer treatment adjuvant to re-boost patient immune responses with variable clinical outcomes. We aimed to clarify the impact of attenuated heat-killed tuberculosis (HKTB) on tumor-associated macrophages which play critical roles in shaping immunological regulation in the tumor microenvironment. Upon HKTB stimulation, both primary macrophages derived from the peripheral blood of healthy subjects and from lung cancer patients as well as THP1-derived classically activated macrophages (Ms) and tumor-educated macrophages (TEMs) were polarized into the proinflammatory phenotype, as characterized by increased expression cluster of differentiation 86. A quantitative proteomic analysis revealed that stimulated TEMs were unable to activate the toll-like receptor 2, signal transducer and activator of transcription 1, or nuclear factor-κB signaling. Instead, they showed distinct intercellular adhesion molecule 1 signaling, impaired cell adhesion, and mitochondrial dysfunction. These molecular mechanisms might contribute to lower cytotoxicity of HKTB-stimulated TEMs against A549 cells via the release of distinct inflammatory cytokines compared to HKTB-stimulated Ms. Our study provides an unbiased and systematic interpretation of cellular and molecular alterations of HKTB-reeducated macrophages which should help illuminate potential strategies of HKTB-stimulated macrophage-based combination therapy for cancer treatment.


Ery Kus Dwianingsih, Junaedy Yunus, Lutfan Lazuardi, Amirah Ellyza Wahdi, Aulia Fitri Rhamadianti, Florentina Linda, Sunandar Hariyanto, Jajah Fachiroh


BACKGROUND: Biobanks play an essential role in the development of personalized medicine since they collect large numbers of high-quality biomaterials corresponding to clinical data. Despite its extensive population diversity, research institutions in Indonesia have indicated less awareness regarding biobanking for research practices.AIM: The journey to harmonize the knowledge and understanding of biobanks for health research and the development of the network in Indonesia has been summarized in this article.

METHODS: To build a national biobank network, in 2015 the Faculty of Medicine, Public Health and Nursing, UGM held the first national biobank network meeting in Indonesia. Follow-up meetings were then held to identify challenges and constraints faced by the network. Five annual national workshops (2015–2019) have been held.

RESULTS: Four working groups (WG) were formed to effectively coordinate the network, addressing the infrastructure and Laboratory Information Management System (WG 1), SOP and Best Practices (WG 2), Training and Education and Legal (WG 3), and Ethical and Social Issues (WG 4).

CONCLUSION: The formation of a national biobank network in Indonesia is based on the hope for multi-institutional collaboration to mainly foster the development of biobanks for health research with best available practices and provide a central hub of coordination.


Ika Fidianingsih, Teguh Aryandono, Sitarina Widyarini, Sri Herwiyanti, Sunarti Sunarti


Background: Breast cancer prevention still needs to be improved. Calorie restriction is thought to prevent breast cancer through the induction of autophagy. Maranta arundinacea L. (MA) has the potential for calorie restriction because it contains high fiber. This research aimed to observe the effect of dietary MA against dimethylbenz(a)anthracene (DMBA)-induced mammary cancer in Sprague Dawley rats related to autophagy. Methods: Twenty-five Sprague Dawley rats were randomly divided into five groups: 1) control group without DMBA-induced with a standard diet, 2) 20 mg/kg BW of DMBA two times a week for five weeks with a standard diet, 3) DMBA and diet modification with 30% of MA, 4) DMBA and diet modification with 45% of MA, and 5) DMBA and diet modification with 60% of MA. Examination of the nodule was conducted once every week for 22 weeks. Breast tissue/tumor examination underwent histology examination with hematoxylin-eosin. Examinations of immunohistochemical staining against Beclin1, LC3B, and SQSTM1 were conducted to reveal autophagy. The difference of autophagy protein expression was analyzed using One way ANOVA with 95% confidence level and significance set as p<0.05. Results: Cancer was detected in four rats of DMBA standard diet, two rats of 30% MA, one rat of 45% MA. No cancer was detected in the rats of control and rats with 60% of MA group. The Beclin1 expressions showed that the 60% of MA group had the
highest score (2.5±0.52) followed by the 45% of MA group (1.87±0.49), control group (1.77±0.11), 30% of MA group
(1.28±0.75), and DMBA with standard diet had the lowest score (1.28±0.91). The difference of Beclin1 expressions was statistically significant (p-value=0.03). However, the difference of the LC3B expressions (p-value=0.11) and SQSTM1 expressions (p-value=0.225) were not statistically significant. Conclusion: Dietary modifications with MA potentially prevent breast cancer and induce initiation of autophagy.
Maranta arundinacea– breast cancer- DMBA- Arrowroot- autophagy

DOI: 10.31557/APJCP.2022.23.3.985

Nastiti Wijayanti, Faris Muhammad Gazali, Endah Supriyati, Mohamad Saifudin Hakim, Eggi Arguni, Marselinus Edwin Widyanto Daniwijaya, Titik Nuryastuti, Matin Nuhamunada, Rahma Nabilla, Sofia Mubarika Haryana, Tri Wibawa


Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is a new virus responsible for the COVID-19 pandemic. The emergence of the new SARS-CoV-2 has been attributed to the possibility of evolutionary dynamics in the furin cleavage site (FCS) region. This study aimed to analyze the sequence of the FCS region in the spike protein of SARS-CoV-2 isolates that circulated in the Special Region of Yogyakarta and Central Java provinces in Indonesia. The RNA solution extracted from nasopharyngeal swab samples of confirmed COVID-19 patients were used and subjected to cDNA synthesis, PCR amplification, sequencing, and analysis of the FCS region. The sequence data from GISAID were also retrieved for further genome analysis. This study included 52 FCS region sequences. Several mutations were identified in the FCS region, i.e., D614G, Q675H, Q677H, S680P, and silent mutation in 235.57 C > T. The most important mutation in the FCS region is D614G. This finding indicated the G614 variant was circulating from May 2020 in those two provinces. Eventually, the G614 variant totally replaced the D614 variant from September 2020. All Indonesian SARS-CoV-2 isolates during this study and those deposited in GISAID showed the formation of five clade clusters from the FCS region, in which the D614 variant is in one specific cluster, and the G614 variant is dispersed into four clusters. The data indicated there is evolutionary advantage of the D614G mutation in the FCS region of the spike protein of SARS-CoV-2 circulating in the Special Region of Yogyakarta and Central Java provinces in Indonesia.


Suci Widhiati, Dewajani Purnomosari, Tri Wibawa, Hardyanto Soebono


The close relationship between the intestine and the skin has been widely stated, seen from gastrointestinal (GI) disorders often accompanied by skin manifestations. Exactly how the gut microbiome is related to skin inflammation and influences the pathophysiology mechanism of skin disorders are still unclear. Many studies have shown a two-way relationship between gut and skin associated with GI health and skin homeostasis and allostasis. This systematic review aimed to explore the associations between the gut microbiome with inflammatory skin disorders, such as acne, psoriasis, atopic dermatitis, and urticaria, and to discover the advanced concept of this relationship. The literature search was limited to any articles published up to December 2020 using PubMed and EBSCOHost. The review followed the PRISMA guidelines for conducting a systematic review. Of the 319 articles screened based on title and abstract, 111 articles underwent full-text screening. Of these, 23 articles met our inclusion criteria, comprising 13 atopic dermatitis (AD), three psoriasis, four acne vulgaris, and four chronic urticaria articles. Acne vulgaris, atopic dermatitis, psoriasis, and chronic urticaria are inflammation skin disorders that were studied recently to ascertain the relationship of these disorders with dysbiosis of the GI microbiome. All acne vulgaris, psoriasis, and chronic urticaria studies stated the association of gut microbiome with skin manifestations. However, the results in atopic dermatitis are still conflicting. Most of the articles agree that Bifidobacterium plays an essential role as anti-inflammation bacteria, and Proteobacteria and Enterobacteria impact inflammation in inflammatory skin disorders.


Rizwan Ahmad, Muhammad Riaz, Mohammed Aldholmi, Muhammad Asif Qureshi, Shahab Uddin, Ajaz Ahmad Bhat, Pratheeshkumar Poyil, Mukhtiar Baig, Jalal Pourahmad, Trivadi Ganesan, Abdul Quaiyoom Khan, Zainab Siddiqui, Maha El-Demellawy, Maryam Gholamalizadeh, Dewajani Purnomosari, Elsayed I Salim, Seyedeh Zahra Mousavi Jarrahi, Jian-ye Zhang, Samad Mohammadnejad, Alireza Mosavi Jarrahi


The journal of APJCP (Asian Pacific Journal of Cancer Prevention) focuses to gather relevant and up-to-date novel information’s related to cancer sciences. The research methodologies and approaches adopted by the researcher are prone to variation which may be desirable in the context of novel scientific findings however, the  reproducibility for these studies needs to be unified and assured. The reproducibility issues are highly concerned when preclinical studies are reported in cancer, for natural products in particular. The natural products and medicinal plants are prone to a wide variation in terms of phytochemistry and phyto-pharmacology, ultimately affecting the end results for cancer studies. Hence the need for specific guidelines to adopt a best-practice in cancer research are utmost essential. The current AIMRDA guidelines aims to develop a consensus-based tool in order to enhance the quality and assure the reproducibility of studies reporting natural products in cancer prevention. A core working committee of the experts developed an initial draft for the guidelines where more focus was kept for the inclusion of specific items not covered
in previous published tools. The initial draft was peer-reviewed, experts-views provided, and improved by a scientific
committee comprising of field research experts, editorial experts of different journals, and academics working in different organization worldwide. The feedback from continuous online meetings, mail communications, and webinars resulted a final draft in the shape of a checklist tool, covering the best practices related to the field of natural products research in cancer prevention and treatment. It is mandatory for the authors to read and follow the AIMRDA tool, and be aware of the good-practices to be followed in cancer research prior to any submission to APJCP. Though the tool is developed based on experts in the field, it needs to be further updated and validated in practice via implementation in the field.
AIMRDA- submission guidelines- cancer- natural products- APJCP

DOI: 10.31557/APJCP.2021.22.12.3735

Neti Nurani, Tunjung Wibowo, Rina Susilowati, Janatin Hastuti, Madarina Julia, Mirjam M. Van Weissenbruch


Background: Compared to their appropriate-for-gestational-age (AGA) peers, small-for-gestational-age (SGA) infants are prone to growth deficits. As the first 6 months of exclusive breastfeeding is generally recommended, it is essential to understand how this intervention might impact SGA infants’ growth. This study aims to assess growth of exclusively breastfed SGA term infants in the first 6 months of life. Methods: A prospective cohort study was conducted on term infants born in Dr. Sardjito General Hospital and two private hospitals in Yogyakarta, Indonesia. SGA was defined as birth weight less than the 10th percentile according to Fenton criteria. Weight, length, and head circumference (HC) were measured at birth and monthly until 6 months old. Results: A total of 39 AGA and 17 SGA term infants who were exclusively breastfed in their first 6 months were included and followed. In SGA compared to AGA, birth weight, length, and HC (mean ± SD) were significantly lower (p < 0.001). During the first 6 months, the SGAs grew in weight and length in parallel with the AGAs. At sixth months of age, the weight and length (mean ± SD) of the SGAs were significantly lower compared to the AGAs (p < 0.001). However, HC (mean ± SD) of SGAs grew significantly faster than the AGAs (p < 0.005). At sixth months of age, there were no significant differences in HC between the two groups (p = 0.824). Conclusions: In the first 6 months, exclusively breastfed SGA term infants, in contrast to weight and length, only show catch up growth in HC, leading to HC comparable to their AGA peers at the age of 6 months. Keywords: Growth, Exclusively breastfed, Small for gestational age, Infant DOI:


Open Access Macedonian Journal of Medical Sciences. 2021 Nov 08; 9(F):523-533.

BACKGROUND: Alzheimer’s Dementia (AD) cases are increasing with the global elderly population. To study the part of the brain affected by AD, animal models for hippocampal degeneration are still necessary to better understand AD pathogenesis and develop treatment and prevention measures. AIM: This study was a systematic review of toxic substance-induced animal models of AD using the Morris Water Maze method in determining hippocampal-related memory impairment. Our aim was reviewing the methods of AD induction using toxic substances in laboratory rodents and evaluating the report of the AD biomarkers reported in the models. METHODS: Data were obtained from articles in the PubMed database, then compiled, categorized, and analyzed. Eighty studies published in the past 5 years were included for analysis. RESULTS AND DISCUSSION: The most widely used method was intracerebroventricular injection of amyloid-β substances. However, some less technically challenging techniques using oral or intraperitoneal administration of other toxic substances also produce successful models. Instead of hippocampal neurodegeneration, many studies detected biomarkers of the AD pathological process while some reported inflammation, oxidative stress, neurotrophic factors, and changes of cholinergic activity. Female animals were underrepresented despite a high incidence of AD in women. CONCLUSION: Toxic substances may be used to develop AD animal models characterized with appropriate AD pathological markers. Characterization of methods with the most easy-handling techniques and more studies in female animal models should be encouraged.

European Journal of Plastic Surgery

Background: Traumatic brain injury (TBI) is one of the leading causes of death and disability worldwide. In various combinations, a high frequency of facial bone fractures has been reported. However, the results are still conflicting. Methods: We retrospectively reviewed medical records of patients with maxillofacial trauma who were admitted to Dr. Sardjito Hospital, Yogyakarta, Indonesia, between January 2016 and December 2017. Results: A total of 70 patients with maxillofacial trauma were involved (57 males and 13 females, 18–65 years). Moreover, most of them were 18–25-year-old males (34.3%). The average patient age was 35.5 ± 14 years. No significant association was found between the sex and age group of the patient (p = 0.774). Motorcycle accident was the most frequent cause of maxillofacial fractures (84%) with midfacial and multiple maxillofacial fractures being the most frequent types found in patients (40% and 40%, respectively). The most common facial fracture was in the zygomatic bone (28%) and most cases showed mild facial injury (81%). A significant association was found between the Glasgow Coma Scale (GCS) and types of traumatic brain injury (TBI) (p = 0.031). However, when GCS was compared with facial injury, no statistically significant level was reached and its correlation was low (p = 0.267, r = 0.134). Conclusions: There was a significant association between types of traumatic brain injury with head injury severity. However, we found no correlation between head injury severity and facial injury severity. To explain and validate our results, further multicenter studies with a larger sample size are required. Level of evidence: Level IV, Risk/Prognostic.

PLoS One. 2021 Oct 15;16(10):e0258617.
doi: 10.1371/journal.pone.0258617. PMID: 34653200; PMCID: PMC8519449.

Background: It has been shown that vitamin D is associated with obesity and the development of atherosclerosis. Less is known about this association among adolescents with obesity. Objectives To determine the association of vitamin D level and metabolic risk factors with carotid intima-media thickness (CIMT) among obese adolescents. Methods: We conducted a cross-sectional study among obese children aged 15 to 17 years in Yogyakarta, Indonesia. The association of vitamin D and other metabolic risk factors (triglyceride, low-density lipoprotein cholesterol (LDL-C) and high-density lipoprotein cholesterol (HDL-C), and insulin resistance using homeostasis model assessment of insulin resistance (HOMA-IR)) with CIMT was explored by multivariable linear regression models. Results: Out of 156 obese adolescents, 55.8% were boys. Compared to girls, boys had higher BMI z-score, waist circumference, and HDL-cholesterol. After adjustment for age, sex and second-hand smoke exposure, high HOMA-IR, total cholesterol, LDL-cholesterol and triglyceride levels were associated with higher odds of elevated CIMT. In analyses stratified by sex, a similar trend was observed in boys, while none of the risk factors were associated with CIMT in girls. We observed no association between vitamin D and CIMT. Conclusions: Hyperinsulinemia, higher total cholesterol and LDL cholesterol were associated with greater odds of elevated CIMT among obese adolescent boys.

Reviews in Cardiovascular Medicine 2021, Vol. 22 Issue (3): 919-924
DOI: 10.31083/j.rcm2203099

Central obesity is associated with increased level and activity of endothelin-1. The waist and hip circumferences are simple indicators of central obesity. Waist circumference correlates with visceral adiposity, whereas hip circumference associates with gluteofemoral peripheral adiposity. Both measurements have independent and opposite correlation with coronary artery disease (CAD) risk factors. The relation between serum endothelin-1 in stable CAD and both parameters of central obesityneeds to be investigated. This study aims to examine the correlation between serum endothelin-1 level and waist and hip circumferences as parameters of central obesity in patients with stable CAD. This was a cross-sectional study. Consecutive subjects were enrolled among those who underwent elective coronary angiography with significant CAD. Serum endothelin-1 was measured from peripheral blood samples taken before coronary angiography procedure. The measurement of waist circumference, hip circumference, and ratio derived from them, was performed. Central obesity was determined by waist circumference cut-off for Indonesian population. The correlation analysis was performed with Pearson test. The multivariate analysis was performed with multiple linear regression test. The comparison of serum endothelin-1 level between groups was performed with Student T test. We enrolled 50 subjects. The majority of subjects was male (80.0%), hypertensive (86.0%), dyslipidemic (68%) and smoker (52%). Most subjects had history of acute coronary syndrome (64%). Mean waist circumference was 87.6 +/- SD cm, hip circumference was 95.3 cm +/- SD, mean waist-to-hip ratio was 0.92 +/- SD and mean waist-to-height ratio was 0.54 +/- SD. Central obesity occurred in 32% of subjects. Mean serum endothelin-1 level was 2.2 ± 0.7 pg/mL. Serum endothelin-1 level tended to be higher in subjects with central obesity as compared to those without. Serum endothelin-1 level was significantly correlated with age, hemoglobin level, waist circumference (coefficient of 0.311, p value = 0.023) and hip circumference (coefficient of 0.359, p value = 0.010). Multivariable analysis indicated that age (coefficient of -0.353, p value = 0.007) and hip circumference (coefficient of 0.335, p value = 0.011) were independently correlated with serum endothelin-1. For conclusion, in patients with stable CAD, serum endothelin-1 was positively correlated with both waist circumference and hip circumference. Hip circumference independently and positively correlated with serum endothelin-1 level.

Sci Rep. 2021 Aug 9;11(1):16077.
DOI: 10.1038/s41598-021-95381-0. PMID: 34373489; PMCID: PMC8352923

While there has been significant progress in the molecular characterization of the childhood brain cancer medulloblastoma, the tumor proteome remains less explored. However, it is important to obtain a complete understanding of medulloblastoma protein biology, since interactions between proteins represent potential new drug targets. Using previously generated phosphoprotein signaling-profiles of a large cohort of primary medulloblastoma, we discovered that phosphorylation of transcription factor CREB strongly correlates with medulloblastoma survival and associates with a differentiation phenotype. We further found that during normal cerebellar development, phosphorylated CREB was selectively expressed in differentiating cerebellar granule neuron progenitor (CGNP) cells. In line, we observed increased differentiation in CGNPs treated with Forskolin, Bmp6 and Bmp12 (Gdf7), which induce CREB phosphorylation. Lastly, we demonstrated that inducing CREB activation via PKA-mediated CREB signaling, but not Bmp/MEK/ERK mediated signalling, enhances medulloblastoma cell sensitivity to chemotherapy.

Sci Rep. 2021 Aug 9;11(1):16077.
doi: 10.1038/s41598-021-95381-0

While there has been significant progress in the molecular characterization of the childhood brain cancer medulloblastoma, the tumor proteome remains less explored. However, it is important to obtain a complete understanding of medulloblastoma protein biology, since interactions between proteins represent potential new drug targets. Using previously generated phosphoprotein signaling‑profiles of a large cohort of primary medulloblastoma, we discovered that phosphorylation of transcription factor CREB strongly correlates with medulloblastoma survival and associates with a differentiation phenotype. We further found that during normal cerebellar development, phosphorylated CREB was selectively expressed in differentiating cerebellar granule neuron progenitor (CGNP) cells. In line, we observed increased differentiation in CGNPs treated with Forskolin, Bmp6 and Bmp12 (Gdf7), which induce CREB phosphorylation. Lastly, we demonstrated that inducing CREB activation via PKA‑mediated CREB signaling, but not Bmp/MEK/ERK mediated signalling, enhances medulloblastoma cell sensitivity to chemotherapy.

Asian Pac J Cancer Prev. 2021 Aug 1;22(8):2363-2370.
doi: 10.31557/APJCP.2021.22.8.2363

Objective: To investigate the correlation between TLR3 and pro-inflammatory cytokines (TNFα, IL6) expression with the distribution of macrophage M2 and Treg on Epstein Barr virus-encoded RNAs (EBER+) nasopharyngeal carcinoma (NPC) tissues. Methods: A total of 23 FFPE NPC tissue samples were obtained from patients in Dr. Sardjito General Hospital, Yogyakarta, Indonesia in 2008–2010, which expressed EBER was collected. The expressions of TLR3, TNFα, and IL6 were examined using an immunofluorescence assay. The distribution of macrophage M2 and Treg were examined by immunohistochemistry with anti-CD163 and -FOXP3 antibodies, respectively. The quantification of fluorescence intensity was analyzed by the RGB space method using ImageJ software. The M2 interpretation was done by the eyeballing method and the M2 scores were divided into 0 (negative), 1 (scant), 2 (focal), 3 (abundant). The average number of Treg FOXP3+ cells in five high power fields was counted. The relationship between variables was tested by the Spearman correlation test, and the coefficient correlation was used to see the correlation between variables. Results: All EBER+ NPC specimens showed TLR3 expression intracellularly. The expression of TNFα could be observed in the cell membranes and secreted extracellularly, while IL6 was secreted to the extracellular area. The expression of TNFα was two times higher than IL6. Most specimens showed low M2 score (56.52%) and high Treg (52.17%). A positive correlation was found between TLR3 and IL6 (12.9%). TNFα was positively correlated with the M2 distribution of 13.7% and Treg distribution of 12.9%, while the rest were explained by other factors. Conclusion: TNFα has a positive correlation with M2 and Treg distribution, but mostly through a different mechanism other than EBER-TLR3 interaction. Possibly, other pro-inflammatory and anti-inflammatory cytokines are involved in the formation of the NPC microenvironment, especially related to the presence of M2 and Treg, which provide immunosuppressive effects in NPC tumors

The Indonesian Biomedical Journal, Vol.13, No.2, June 2021, p.106-220
DOI: 10.18585/inabj.v13i2.1463

BACKGROUND: Single nucleotide variations (SNV) have been mapped to be associated with several human conditions and diseases. To validate the association between SNV to certain human traits or diseases, a large number of subjects must be included. Thus, in need of a fast, relatively economic, and reliable genotyping method. This can be achieved through the use of tetra-primer amplification refractory mutation system polymerase chain reaction (Tetra-primer ARMS PCR). This study reports strategy to develop Tetra-primer ARMS PCR-based genotyping of CHRNA3 rs8040868. METHODS: The optimization of Tetra-primer ARMS PCR was done through these steps: identification of gene sequence and position of single mutation; designing outer and inner PCR primers; amplification of target gene fragments through PCR by using outer primer; confirming genotype of the PCR product by using sequencing; determining an optimum ratio of outer and inner primer; and determining optimum annealing temperature and cycles for the PCR program. The PCR products were run in 2% gel agarose electrophoresis and visualized under UV illumination. RESULTS: Outer and inner primer ratio of 1:3 with annealing temperature of 64.4oC and 40x cycles was found to be the most optimum condition. Tetra-primer ARMS PCR was able to confirm the results of the DNA sequence of 2 samples, confirming wild-type variants (TT allele) and the heterozygous mutant (CT allele). CONCLUSION: Tetra-primer ARMS PCR was able to genotype rs8040868 of the CHRNA3 gene.

Intractable & Rare Diseases Research. 2021; 10(2):88-94.
DOI: 10.5582/irdr.2020.03150

Epidermolysis bullosa (EB) is a group of inherited blistering skin diseases known to have heterogenicity of phenotypes and genotypes. There are four main types of EB: Simplex, junctional, dystrophic, and Kindler syndrome, which are further classified into 34 distinct subtypes. Twenty different gene mutations are responsible for the loss of function and integrity of the basal membrane zone. In limited-resource settings such as Indonesia, diagnoses of hereditary skin disease often rely on clinical features. This limitation was managed by using the Clinical Diagnostic Matrix EB for clinical diagnosis support and whole-exome sequencing for genetic analysis. This study is the first whole-exome sequencing analysis of Javanese Indonesian patients with EB. The genetic analysis from four patients with EB identified all novel mutations unreported in the dbSNP database. There is Kindler syndrome with FERMT1 frameshift mutation in exon 4, at c.388A (p.I130fs), which causes truncated protein; junctional EB generalized intermediate (JEB-GI) subtype with missense mutation at LAMB3 gene position c.A962C (p.H321P); and recessive dystrophic EB (RDEB) a missense mutation at COL7A1 gene position c.G5000T (p.G1667V). The whole-exome sequencing was further verified by Sanger sequencing. The new mutations’ finding is possibly due to the limited genetic database in the Malayo-Polynesian ethnic group. Indonesia has hundreds of ethnic groups, and the Javanese is the largest ethnic group that populates Indonesia. Genetic data of these ethnic groups is important to be established in the international genetic database. This combination of clinical diagnostic and genetic analysis tools with whole-exome sequencing confirmed the challenging diagnosis of epidermolysis bullosa.

Front Pediatr. 2021 May 25;9:680869.
doi: 10.3389/fped.2021.680869. PMID: 34113592; PMCID: PMC8185158.

Background: Interactions between the genome and intrauterine environment can affect bone mineralization in newborns and even in adult life. Several studies show that intrauterine fetal bone mineralization or early postnatal bone condition influences the risk of osteoporosis in later life. Objectives: To determine whole body bone mineral content (WB BMC) and factors that influence neonatal WB BMC in Indonesian term newborns. Subjects/Methods: A cross-sectional study was conducted in Dr. Sardjito General Hospital, Yogyakarta, Indonesia. A total of 45 term, appropriate for gestational age (AGA) newborns were included in this study. BMC was assessed by dual-energy x-ray absorptiometry (DXA) in the first week of life. Weight (g), length (cm) and head circumference (cm) were measured at birth. Data on maternal characteristics were obtained from the maternal health records or reported by the mothers. Results: WB BMC measured in the present study (mean ± SD: 33.2 ± 9.3 g) was lower than WB BMC of similar populations in developed countries. Multiple linear regression showed that birth weight, birth length, and gestational age had a positive association with WB BMC (p = 0.048, 0.017, and <0.001, respectively), while maternal cigarette exposure had a negative association with WB BMC (p = 0.012). Male infants had significantly higher of WB BMC than female (p = 0.025). These determinants contribute to 55% variability of WB BMC. Conclusions: WB BMC in Indonesian term newborns is lower than populations in developed countries. Birth weight, length, gestational age, sex, and maternal cigarette exposure during pregnancy are significantly associated with WB BMC observed in Indonesian newborns.

Asian Pacific Journal of Cancer Prevention. Volume 22, Issue 11, Pages 3447 – 3453
DOI: 10.31557/APJCP.2021.22.11.3447

Objective: This study aimed to identify the distribution of M2 macrophage and Treg in Nasopharyngeal Carcinoma (NPC) tumor tissue samples. The presence of these two groups of cells was further correlated to clinical stage, tumor size, the lymphatic node involvement, and metastasis. Methods: The total of 50 formalin-fixed paraffin-embedded (FFPE) NPC tissue samples was collected retrospectively (27 samples) and prospectively (23 samples). Samples were FFPE tissue slices. Immunohistochemistry was done on the FFPE tissue slides using anti-CD-163 and anti-FoxP-3 antibodies for M2 macrophage and Treg detection, respectively. The M2 macrophage interpretation was performed by eye-balling method and the score was divided into 0 (negative), 1 (scant), 2 (focal), and 3 (abundant). The average number of Treg FOXP3+ cells in 5 high power fields (HPF) was calculated. The relationship of M2 macrophage and Treg was tested with Spearman’s correlation. The relationship between M2 macrophage and Treg with clinical stage, tumor size, node involvement and metastasis was tested by chi square, with p<0.1. Results: M2 macrophage and Treg were positive correlated (r=0.469, p<0.001). The presence of M2 macrophage and regulatory T cell (Treg) was significantly correlated to tumor size (p= 0.091 for M2 macrophage and p=0.022 for Treg) and clinical stage (p= 0.030 for M2 macrophage and p= 0.002 for Treg), but did not correlate with lymphatic node involvement and metastasis. Conclusions: In Epstein-Barr virus related NPC tumor microenvironment, the presence of M2 macrophage was correlated with Treg, and both types of the cells were correlated with tumor size and clinical stages.

Septyaningtrias DE, Susilowati R. Neurological Involvement of COVID-19: from Neuroinvasion and Neuroimmune Crosstalk to Long-term Consequences. Rev Neurosci. 2021 Feb 1;32(4):427-442.  doi: 10.1515/revneuro-2020-0092.

As the coronavirus disease 2019 (COVID-19) pandemic continues to be a multidimensional threat to humanity, more evidence of neurological involvement associated with it has emerged. Neuroimmune interaction may prove to be important not only in the pathogenesis of neurological manifestations but also to prevent systemic hyperinflammation. In this review, we summarize reports of COVID-19 cases with neurological involvement, followed by discussion of possible routes of entry, immune responses against coronavirus infection in the central nervous system and mechanisms of nerve degeneration due to viral infection and immune responses. Possible mechanisms for neuroprotection and virus-associated neurological consequences are also discussed.