Prognostic Value for Mortality of Plasma Bioactive Adrenomedullin in Patients with Pulmonary Arterial Hypertension: A Sub Analysis of the Biomarker Study in the COHARD-PH Registry.

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.

Increased endothelin-1 levels in coronary artery disease with diabetes mellitus in an Indonesian population.

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.

Serum endothelin-1 level positively correlates with waist and hip circumferences in stable coronary artery disease patients.

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.

Origin and distribution of nitrate in water well of settlement areas in Yogyakarta, Indonesia.

Pollution of nitrate in water wells in Yogyakarta City was reported to increase for two decades. This study aimed to describe nitrate contamination in the water wells of colorectal cancer (CRC) and inflammatory bowel disease (IBD) patients, previously described elsewhere. Nitrate and chloride content of 150 water samples from the wells of patients with CRC and IBD who were residing in Yogyakarta, Sleman, or Bantul districts were examined. Description of nitrate contamination was presented in the form of box plot charts and map. Kruskal-Wallis analysis was used to measure the difference of nitrate concentration in three areas of study. Comparisons of nitrate and chloride concentrations were used to determine the source of nitrate contamination in water well. Fisher's exact test was used to describe the relationship of well distance with the septic tank to nitrate concentration in water well. The wells in Yogyakarta City had the highest median nitrate content compared to Sleman and Bantul (P = 0.001) with the median of 56.6, 13.1, and 7.7 for Yogyakarta, Sleman, and Bantul, respectively, and most tested samples exceed WHO safe drinking water standards. The spread of nitrate contamination has occurred in areas adjacent to Yogyakarta City compared to the previous report. The ratio of nitrate to chloride (1-8:1) suggested that the source of nitrate contamination in water wells in the study area came from feces due to inadequate on-site sanitation. The mapping showed nitrate contamination in water wells in Yogyakarta City, Sleman, and Bantul districts had spread according to urban development.

Nitrate in drinking water and risk of colorectal cancer in Yogyakarta, Indonesia.

Nitrate concentration in well water in Yogyakarta, Indonesia, and its surroundings tended to increase rapidly from time to time, and it may be associated with an elevated risk for several types of cancer. The purpose of this study was to examine the association between nitrate in drinking water and colorectal cancer (CRC) risk occurrence. A case-control study was conducted in Yogyakarta Special Province. Pathologically confirmed 75 CRC patients and 75 controls were consulted and their individual well water was sampled and examined for nitrate concentrations. Logistic regression analysis was conducted to establish the association between nitrate and CRC risk development. There was a significant correlation between nitrate in drinking water and CRC occurrence, and this value was relatively stable after being adjusted for protein intake, smoking history, age, and family history of cancer. These findings demonstrated that the risk of CRC development was fourfold among those with >10 years of nitrate exposure from well water compared with those with ≤10 years of nitrate exposure. Consequently, a significant association between nitrate in drinking water and occurrence of CRC in Yogyakarta was established.

Diverging likelihood of colon and rectal cancer in Yogyakarta, Indonesia: A cross sectional study.

OBJECTIVES: Colon and rectal cancer are associated with different risk factors and prognostic. However, this discrepancy has not been widely explored in the local population. This study aimed to investigate the site-specific likelihood of colorectal cancer (CRC) incidence in the Yogyakarta province, Indonesia. METHODS: This cross-sectional study analyses 1,295 CRC cases diagnosed in 2008-2019 registered in the Yogyakarta population-based cancer registry (PBCR) database. Cases were grouped into colon and rectal cancer. Log-binomial regression was used to determine the relative risk of either colon or rectal cancer across different gender, age group, and rurality of residence. The age-specific rates were calculated by age group and temporal trend for each group were analyzed using joinpoint regression. RESULTS: Females displayed higher odds of colon cancer (relative risk/RR = 1.20, 95%CI = 1.02-1.41) and lower odds of rectal cancer (RR = 0.92, 95%CI = 0.85-0.99). Elevated odds of colon cancer were observed in younger age group, especially 30-39 (RR = 1.87, 95%CI = 1.10-3.19), while decreased odds of rectal cancer was apparent in age group 30-39 and 40-49 (RR = 0.75, 95%CI = 0.60-0.93 and RR = 0.82, 95%CI = 0.69-0.98, respectively). Living in urban or rural areas did not significantly influence the odds of either having colon (RR = 0.98, 95%CI = 0.82-1.17) or rectal cancer (RR = 1.01, 95%CI = 0.93-1.10). During 2008-2019, trends of colon cancer in age <50 increased by 8.15% annually while rectal cancer displayed a 9.71% increase annually prior to 2017, followed by a 17.23% decrease until 2019. CONCLUSIONS: Yogyakarta population shows higher odds of young-onset colon cancer, especially between age 30-39 years old. Overall observation of trend shows increasing incidence in young-onset colon cancer, and non-significant decrease in rectal cancer.

Improving Public Trust in Biobanking: Roundtable Discussions from the 2021 ISBER Annual Meeting.

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.

Temporal and spatial analyses of colorectal cancer incidence in Yogyakarta, Indonesia: a cross-sectional study.

We aimed to explore the district-level temporal dynamics and sub-district level geographical variations of colorectal cancer (CRC) incidence in the Special Region of Yogyakarta Province. We performed a cross-sectional study using data from the Yogyakarta population-based cancer registry (PBCR) comprised of 1,593 CRC cases diagnosed in 2008-2019. The age-standardized rates (ASRs) were determined using 2014 population data. The temporal trend and geographical distribution of cases were analysed using joinpoint regression and Moran's I statistics. During 2008-2019, CRC incidence increased by 13.44% annually. Joinpoints were identified in 2014 and 2017, which were also the periods when annual percentage change (APC) was the highest throughout the observation periods (18.84). Significant APC changes were observed in all districts, with the highest in Kota Yogyakarta (15.57). The ASR of CRC incidence per 100,000 person- years was 7.03 in Sleman, 9.20 in Kota Yogyakarta, and 7.07 in Bantul district. We found a regional variation of CRC ASR with a concentrated pattern of hotspots in the central sub-districts of the catchment areas and a significant positive spatial autocorrelation of CRC incidence rates in the province (I=0.581, p<0.001). The analysis identified four high-high clusters sub-districts in the central catchment areas. This is the first Indonesian study reported from PBCR data, showing an increased annual CRC incidence during an extensive observation period in the Yogyakarta region. A heterogeneous distribution map of CRC incidence is included. These findings may serve as basis for CRC screening implementation and healthcare services improvement.

Tobacco consumption and genetic susceptibility to nasopharyngeal carcinoma (NPC) in Thailand.

BACKGROUND: The incidence of nasopharyngeal carcinoma (NPC) varies substantially worldwide, with an endemic pocket in Southeast Asia. METHOD: We assessed lifestyle and genetic factors in relation to NPC risk among 681 NPC cases and 1,078 controls from Thailand. Evaluated lifestyle factors included traditionally preserved foods, tobacco smoking, betel quid chewing, and alcohol consumption. Genetic factors included six variants implicated in a previous a genome-wide association study (GWAS) of NPC and three variants residing near the CHRNA3 and TERT genes that were linked to lung cancer risk in Asian populations. Odds ratios (OR) and 95 % confidence intervals (95 % CI) were estimated using unconditional logistic regression. RESULTS: Frequent consumption of fermented vegetables was associated with increased NPC risk (OR of consumption ≥weekly vs. ≤rare 1.78, 95 % CI 1.24-2.55, p (trend) = 0.005), as was tobacco smoking (p (trend) < 0.001), former and current smokers displaying OR of 1.57 (95 % CI 1.10-2.30) and 2.00 (95 % CI 1.48-2.71) compared to never smokers, respectively. Four out of six genetic variants implicated in the recent NPC GWAS were associated with NPC risk (p (trend) ≤ 0.03), as well as two variants (rs402710 and rs2736098) on the TERT locus at 5p15.33 (p = 0.004 and p = 0.04, respectively). CONCLUSIONS: These results strengthen our previous observation that tobacco smoking is an important risk factor of NPC in this population. Four out of six genetic variants identified in a recent NPC GWAS were confirmed, and the association noted with variants on 5p15.33 suggests that this locus is involved in NPC susceptibility, representing a novel finding in NPC epidemiology.

Seroreactivity against Epstein-Barr virus (EBV) among first-degree relatives of sporadic EBV-associated nasopharyngeal carcinoma in Indonesia.

Epstein-Barr virus (EBV) infection and family history are significant risk factors associated with undifferentiated nasopharyngeal carcinoma. The presence of aberrant immunoglobulin A (IgA) antibodies against specific EBV antigens in healthy individuals can be predictive of the disease. Very limited reports explored the EBV IgA antibody presence within families of sporadic cases of nasopharyngeal carcinoma. This study aimed to determine whether EBV IgA was observed more frequently among family members of sporadic cases of nasopharyngeal carcinoma compared to community controls and evaluated the non-viral factors as determinants of antibody level. First-degree relatives of nasopharyngeal carcinoma patients (n = 520) and case-matched community controls (n = 86) were recruited. Sera from all individuals were tested in standardized peptide-based EBV IgA ELISA. Data on demographic variables and other exogenous factors were collected using a questionnaire through face-to-face interviews. A similar frequency of EBV IgA (cut-off value/CoV 0.354) was observed in the first-degree relatives of cases and in community controls (41.2% vs. 39.5%, P = 0.770). However, with a higher antibody level (OD(450) = 1.000; about three times standard CoV), the relatives showed significantly higher frequency (36.9% vs. 14.7%, P = 0.011). When adjusted for all exogenous factors, the strongest factors associated with seropositivity are being a father (odds ratio/OR = 4.36; 95% confidence interval/CI = 1.56-12.21) or a sibling (OR = 1.89; 95% CI = 1.06-3.38) of a case of nasopharyngeal carcinoma. The higher level of EBV IgA seroreactivity in first-degree relatives of sporadic cases of nasopharyngeal carcinoma compared to the general population supports the use of EBV IgA ELISA for screening among family members.

Preparation of the "Lexique" for ISBER Best Practices 4th Edition for Biobankers in Indo-Pacific Rim Region.

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.

Allopurinol-Induced Stevens-Johnson Syndrome in Javanese Men With Positive HLA-B*58:01.

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.

Decarbonization in Biobanking: A Potential New Scientific Area.

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.

Two-years antibody responses following SARS-CoV-2 infection in humans: A study protocol.

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.

Humoral immune responses to Epstein-Barr virus encoded tumor associated proteins and their putative extracellular domains in nasopharyngeal carcinoma patients and regional controls.

Epstein-Barr virus (EBV) latency proteins EBNA1, LMP1, LMP2, and BARF1 are expressed in tumor cells of nasopharyngeal carcinoma (NPC). IgG and IgA antibody responses to these non-self tumor antigens were analyzed in NPC patients (n=125) and regional controls (n=100) by three approaches, focusing on the putative LMP1, LMP2 extracellular domains. Despite abundant IgG and IgA antibody responses to lytic antigens and EBNA1, patients had low titer (1:25-1:100) IgG to LMP1 (81.2%), LMP2 (95.6%), and BARF1 (84.8%), while immunoblot showed such reactivity in 24.2%, 12.5%, and 12.5% at 1:50 dilution, respectively. Few IgA responses were detected, except for EBNA1. Controls only showed IgG to EBNA1. ELISA using peptides from different domains of LMP1, LMP2, and BARF1 also yielded mostly negative results. When existing, low level IgG to intracellular C-terminus of LMP1 (62.9%) prevailed. Rabbit immunization with peptides representing extracellular (loop) domains yielded loop-specific antibodies serving as positive control. Importantly, these rabbit antibodies stained specifically extracellular domains of LMP1 and LMP2 on viable cells and mediated complement-driven cytolysis. Rabbit anti-LMP1 loop-1 and -3 killed 50.4% and 59.4% of X50/7 and 35.0% and 35.9% of RAJI cells, respectively, and 22% of both lines were lysed by anti-LMP2 loop-2 or -5 antibodies. This demonstrates that (extracellular domains of) EBV-encoded tumor antigens are marginally immunogenic for humoral immune responses. However, peptide-specific immunization may generate such antibodies, which can mediate cell killing via complement activation. This opens options for peptide-based tumor vaccination in patients carrying EBV latency type II tumors such as NPC.

The Distribution of M2 Macrophage and Treg in Nasopharyngeal Carcinoma Tumor Tissue and the Correlation with TNM Status and Clinical Stage.

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.

Necrosis Factor-α (TNF-α) and the Presence of Macrophage M2 and T Regulatory Cells in Nasopharyngeal Carcinoma.

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 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 were 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. 
.

Changes of lactate dehydrogenase in corneal edema after cataract surgery treated with trans-corneal oxygenation therapy.

AIM: To investigate the changes in levels of the lactate dehydrogenase (LDH) enzyme in corneal edema after cataract surgery with trans-corneal oxygenation therapy. METHODS: This pre-post design study design conducted on 15 patients with corneal edema after cataract surgery and receiving trans-corneal oxygenation therapy. Tear sample (using Schirmer paper, from the inferior fornix of the conjunctiva) was carried out prior to trans-corneal oxygenation therapy, on the day 2 (D2) and day 5 (D5) postoperatively before and after trans-corneal oxygenation therapy. Visual acuity [VA (LogMAR)], corneal endothelial density, central corneal thickness (CCT), and coefficient of variation corneal endothelial (CoV) were recorded. The value of LDH was measured using ELISA. The difference in mean LDH value before and after trans-corneal oxygenation therapy, between two groups were analyzed using Wilcoxon signed rank test. RESULTS: There was a decrease in LDH tear concentration at D2 (pre vs post: 1127.54±497.09 vs 696.91±489.49; P=0.002) and D5 (pre vs post: 1064.17±677.77 vs 780.28±428.95; P=0.027) after trans-corneal oxygenation therapy as well as decrease in LDH concentration on the D2 compared to D5 (P=0.041). The mean CCT was decreased significantly after the administration of trans-corneal oxygenation (pre vs post: 632.10±25.66 vs 563.90±51.54; P=0.005). The mean VA and CoV increased significantly after the administration of trans-corneal oxygenation (P=0.001 and P=0.028, respectively). However, there was no difference in mean of corneal endothelial density (P=0.814). CONCLUSION: Trans-corneal oxygenation therapy is associated with significant decrease of tears LDH levels in post cataract surgery with corneal edema. It is accompanied by clinical improvement such as significant reduction of CCT.

Diverse Responses of the Biobanks in Indo-Pacific Rim Region During the COVID-19 Pandemic: Case Scenarios from Two Low- and Middle-Income Countries and Two High-Income Countries in the Indo-Pacific Rim Region.

Background: Biobankers have been unexpectedly involved in the pandemic of COVID-19 since early 2020. Although specific guidance was not available, the International Society for Biological and Environmental Repositories (ISBER) Best Practices and the ISO 20387 document have been utilized to deal with the pandemic disaster. The ISO experts and best practice experts in ISBER teamed up to share the available information and learn the experiences of biobanks concerning COVID-19 through organizing webinars, surveys, and town hall meetings. Four ISBER regional ambassadors (RAs) from the Indo-Pacific Rim (IPR) region were also actively involved at one of the town hall meetings. These RAs, who are from Australia, India, Indonesia, and Japan, and the Director-at-Large of the region, have summarized their experiences in this article. Materials and Methods: The ISBER Standards Committee COVID-19 Task Force has kindly provided the survey results. The extracted glossary from the results was categorized into 10 factors: (1) crisis management; (2) sample-related issues; (3) logistics-related issues; (4) equipment-related issues; (5) ethical, legal, and social implication-related issues; (6) operation-related issues; (7) personnel-related issues; (8) management-related issues; (9) infection-related issues; and (10) research-related issues. Each IPR RA has provided a case considering these 10 factors. Results and Discussion: Two key points have emerged from the scenarios, which are as follows: (1) impacts of the biobanks in low- and middle-income countries (LMICs) are similar to those in high-income countries (HICs) and (2) tolerance of the biobanks in LMICs is not so robust as those in HICs. Furthermore, communication strategies with internal and external stakeholders are critical for a biobank to manage this crisis. This article summarizes the impacts, indicates the opportunities that COVID-19 has brought to the biobank community, and highlights the usefulness of the network beyond biobank services. Lastly, the biobanks need to turn the challenges into opportunities to overcome the crisis.

Tissue P16 is Associated with Smoking Status among Indonesian Nasopharyngeal Carcinoma Subjects.

Background: Nasopharyngeal carcinoma (NPC) is a malignancy with high incidence in Southern China and South-East Asia. NPC incidence among males in Indonesia is estimated around 8.3/100,000 populations. Tobacco smoking is a common risk factor for cancer, including NPC. P16 is one of the key proteins related to the activation of apoptotic pathways, that commonly change during carcinogenesis. Carcinogenesis is often related to environmental exposure, including tobacco smoke. Objective: To analyze the association between P16 protein and smoking status among NPC subjects in Indonesia. Methods: Forty formalin fixed-paraffin embedded NPC tissue samples of known smoking status (20 smokers, 20 non-smokers) were collected from the Department of Anatomical Pathology, Dr. Sardjito Hospital, Yogyakarta. P16 was detected by immunohistochemistry staining. German semi-quantitative scoring system was applied to the P16 staining. Expression index with the score of 0 to 3 was classified as negative staining, meanwhile 4 to 12 was classified as positive staining. The association between P16 (score) and smoking status among NPC patients was analyzed using Fischer exact test. One-sided p ≤ 0.05 was considered as statistically significant. Results: All samples were Javanese males, with age range 25-76 years old. P16 positive staining among smokers was 5% (1/20), while among non-smokers was 40% (8/20). P16 among smokers was significantly lower than non-smokers patients (p=0.010). No difference was found between quantity of smoke and P16 score. Conclusion : A significant association between P16 and smoking status in Indonesian NPC patients has been revealed. The result of this study may be used to improve prevention and management of NPC cases related to smoking habit in Indonesia.