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Objectives:  Combining a three-dimensional scaffold with growth factors before implantation is one method used to increase scaffold bioactivity in bone tissue engineering. The mesenchymal stem cell (MSC)-conditioned medium (CM), called secretome, contains many proteins and growth factors required for tissue repair and growth. This study evaluated the bioactivity of a bovine bone scaffold combined with the secretome of human umbilical cord MSCs (hUC-MSCs) by analyzing MC3T3-E1 cell adhesion and viability on the scaffold., Materials and Methods:  This in vitro laboratory study evaluated the effect of hUC-MSC secretome applied to bovine bone scaffolds processed using various techniques on MC3T3-E1 cell adhesion and viability. The three experimental groups included deproteinized bovine bone mineral-secretome (DBBM-CM), freeze-dried bovine bone-secretome (FDBB-CM), and decellularized FDBB-CM, whereas the control group was treated with DBBM alone. The cell adhesion test was performed using the centrifugation method after 6 and 24 hours, whereas the cell viability test was conducted using the trypan blue exclusion method after 24, 48, and 72 hours. Cell attachment was visualized after 4',6-diamidino-2-phenylindole staining and viewed under inverted fluorescence microscopy., Stastical Analysis:  Statistical analyses were performed using one-way analysis of variance, followed by a post hoc test in cases of significant differences., Results:  Statistical analyses showed significantly greater adhesion of the preosteoblasts to the FDBB-CM scaffold at 6 hours ( p  = 0.002). The results of the adhesion test at 24 hours and the viability tests at all observation times showed no significant differences ( p  > 0.05). This study found that the average MC3T3-E1 cell adhesions and viabilities were highest for the FDBB-CM and DBBM-CM scaffolds. DBBM scaffolds with the secretome had better cell adhesion and viability than those without the secretome., Conclusion:  The addition of MSC secretome increased bovine bone scaffold bioactivity especially in DBBM and FDBB scaffolds., Competing Interests: None declared., (The Author(s). This is an open access article published by Thieme under the terms of the Creative Commons Attribution License, permitting unrestricted use, distribution, and reproduction so long as the original work is properly cited. (https://creativecommons.org/licenses/by/4.0/).)

Background: This study describes the seroconversion and serodynamics of IgG antibodies against the RBD of SARS-CoV-2 in the general population of Sleman District, Yogyakarta Special Province. We aim to identify possible factors that correlate with the seroconversion and serodynamics of IgG antibodies against the RBD of SARS-CoV-2., Methods: We performed a longitudinal study of the population at Health and Demographic Surveillance System (HDSS) Sleman, Yogyakarta, Indonesia. Study subjects were recruited between April and December 2021 using convenience sampling and were followed up 2 times, i.e. 4-5 and 8-9 weeks. The inclusion criteria for subjects were age ≥ 18 years, absence of flu-like symptoms, and negative COVID-19 by using GeNose C19® screening. A community-based survey on demographics, comorbidities and smoking habits were documented at baseline, while a history of vaccination, COVID-19-related symptoms, mobility, and preventive measures, weight and height as well as a venous blood draw, were collected at each visit. The anti-RBD-SARS-CoV-2 IgG antibody concentration from blood plasma was measured using chemiluminescent microplate immunoassay (CMIA). Descriptive analysis was performed based on IgG seropositivity by using chi-squared test or Fisher's exact test, as appropriate. Logistic regression was subsequently performed to identify factors that were correlated with IgG seropositivity. Further, a grouping of subjects based on IgG seropositivity was done to analyze factors that might correlate with seroconversion and serodynamics of anti-RBD-SARS-CoV-2 IgG antibody. A P value ≤ 0.05 was considered to indicate a significant difference., Results: Three hundred eighty-five (385) participants were analyzed. At baseline, 307 out of 385 (79.7%) subjects were seropositive for the IgG antibody against the RBD of SARS-CoV-2. Descriptive analysis showed that sex, marital status, smoking habits, obesity, vaccination status, and preventive measures were different between the IgG anti-RBD-SARS-CoV-2 seropositive and negative individuals (p≤ 0.05). Further analysis showed that, vaccination was the factor most strongly correlated with seropositivity [OR = 20.58; 95% CI 10.82, 39.15]. Based on the correlation, we separated subjects into 4 groups. Group 1 (seronegative-unvaccinated individuals; 50 subjects); Group 2 (seronegative-vaccinated individuals; 27 subjects); Group 3 (seropositive-unvaccinated individuals; 25 subjects); and Group 4 (seropositive-vaccinated individuals; 282 subjects). During monitoring, 27/49 (55.10%), 5/25 (20%), 9/22 (40.91%), and 27/257 (10.51%) of subjects in Group 1, 2, 3, and 4 respectively, received 1 or 2 doses of COVID19 vaccine. When comparing seroconversion at baseline and monitoring 2, positive IgG seroconversion was observed in Group 1 (from 0/51 (0%) to 23/49 (46.94%)) and Group 2 (from 0/27 (0%) to 10/25 (40%)), but negative seroconversion was observed in Group 4 (from 282/0 (100%) to 248/257 (96.50%)); while, all subjects in Group 3 remained seropositive at the end of monitoring. This evidence suggested for hybrid immunity, on which infection and vaccine simultaneously contributes to anti-RBD-SARS-CoV-2 IgG seroconversion., Conclusions: A high seroprevalence of the IgG antibody against RBD-SARS-CoV-2 in the Sleman population was found to correlate with COVID-19 vaccination and as infection occurred, thus enhancing hybrid immunity. We also identified nonresponder and rapid antibody decaying individuals, that call for targeted vaccinations in addition to annual universal boosting., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2025 Fachiroh et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Background: Chemotherapy has a substantial role in decreasing the risk of recurrence and mortality in breast cancer (BC) in a dose-dependent manner where a low relative dose intensity (RDI) is associated with unfavorable outcomes. Several baseline clinicopathological factors, including pro-inflammatory biomarkers, were found to be significant determinants of low RDI. This study aimed to explore the occurrence of low RDI and its influencing factors in women with BC., Methods: This cross-sectional study recruited 172 women with stage I-IV BC who received first-line chemotherapy. We collected patients' clinical, pathological, and treatment data and analyzed the pre-chemotherapy C-reactive protein (CRP) and interleukin (IL)-6 levels using a quantitative enzyme-linked immunosorbent assay (ELISA). We calculated the RDI based on the actual and planned delivered chemotherapy dose (mg/m 2 ) and duration (weeks). RDI less than 85% was defined as "low". Multivariate analysis with logistic regression was conducted to determine the association between pre-chemotherapy parameters and RDI < 85%., Results: The mean CRP level was 10.82 ± 19.17 mg/L (0.00 - 151.73 mg/L) and the mean IL-6 level was 1.12 ± 3.41 pg/mL (0.00 - 27.67 pg/mL). The average RDI for all patients was 93±8.19%. An RDI < 85% occurred in 23 patients (13.4%). The presence of diabetes mellitus (odds ratio (OR): 4.78, 95% confidence interval (CI): 1.03 - 22.27, P = 0.046), triple-negative tumors (OR: 6.45, 95% CI: 1.39 - 29.83, P = 0.017), and IL-6 levels > 0.5 pg/mL (OR: 3.45, 95% CI: 1.01 - 11.79, P = 0.049) was associated with an increased low RDI risk., Conclusion: The proportion of BC patients receiving a low chemotherapy RDI in our study was comparable to published literature and drove close monitoring of patients at risk to provide adequate management., Competing Interests: The authors declare no conflict of interest., (Copyright 2024, Hutajulu et al.)

Purpose: Chemotherapy-induced peripheral neuropathy (CIPN) poses a substantial challenge in breast cancer (BC) chemotherapy, affecting the patient's quality of life. Recent studies have focused on exploring predictors and patterns of CIPN occurrence. We aimed to develop a prediction model for CIPN occurrence using a classification and regression tree (CART) algorithm., Methods: In this prospective study of 170 patients with BC undergoing chemotherapy, patient-reported adaptation of the Common Terminology Criteria for Adverse Events version 4.0 was used to assess CIPN occurrence. Multivariable analysis using the CART model was tuned using 10-fold cross-validation to identify baseline predictors for CIPN throughout chemotherapy. A receiver operating characteristic curve analysis was conducted for the CART model. A multivariable logistic regression was conducted from the variables included in the CART model to assess the strength and direction of the association., Results: The prevalence of CIPN was 64.7% (n = 110). The most decisive predictor of CIPN occurrence in the CART model was the subject's C-reactive protein (CRP) level. CRP level >3.91 mg/dL, BMI >21.85 kg/m 2 , and a marital status of unmarried have predicted a probability of 100% in CIPN occurrence. The CART model showed an accuracy of 65.9%, sensitivity of 51.7%, specificity of 73.2%, and an area under the curve of 0.705. A CRP level of >3.91 mg/dL and a neutrophil-to-lymphocyte ratio (NLR) of >2.82 are significantly associated with the occurrence of CIPN (odds ratio [OR], 2.01 [95% CI, 1.01 to 4.01]; P = .046, OR, 2.09 [95%, CI, 1.02 to 4.24]; P = .042, respectively)., Conclusion: Baseline CRP, NLR, BMI level, and marital status are significant predictors of CIPN occurrence throughout chemotherapy. Our CART model was better at ruling out individuals who would not experience CIPN. The CART model may provide insight into the future development of individualized patient care and prevention strategies.

Inherited retinal dystrophy (IRD) is a group of phenotypes caused by mutations in visual pathways-related genes, mostly occurring at photoreceptors. This heterogeneous group includes retinitis pigmentosa (RP) recognised by bone spicule at the peripheral retina and the other is Stargardt with macular pisiform flecks. In this study, a 20- year-old male patient with RP symptoms was accompanied by a yellowish pisiform flex in the macula. However, his brother, mother and aunty have typical Stargardt disease. This study involved four persons, two males (cases 1 and 2), their mother (case 3) and aunt (case 4). Initially, cases 1 and 2 came to the clinic, case 1 was diagnosed as RP and macular dystrophy, and case 2 was diagnosed as Stargardt disease. On the follow-up, cases 1 and 2 as well as their father, mother and other family members underwent comprehensive eye examination, including fundus, Snellen, OCT, OCT-A and HFA, and found an uncommon macular abnormality besides typical RP appearance in case 1. The father is healthy while the mother and one of his aunties were diagnosed as Stargardt. A genetics analysis was conducted in case 1, finding various mutations associated with IRD mutation at the cone protein-encoded gene that concentrated at the central and rod protein-encoded gene concentrated at the peripheral retina. Whether the combination of multiple or the same mutations is responsible for this RP phenotype needs further analysis and validation. Cases 2 and 3 genetic analysis showed similar mutation results but with a healthy peripheral retina and only represented Stargardt. Case 1 is considered as RP with macular dystrophy, while cases 2, 3 and 4 are confirmed as Stargardt.

Objective: This study examined the morphological changes in the colonic mucosa and the presence of inflammation in rats induced with 1,2-dimethylhydrazine (DMH) 30 mg/kg BW over 9, 11, and 13 weeks without a latency period., Methods: Hematoxylin and eosin staining was performed to assess the morphology and characteristic alteration of the epitheliocytes in the colon. Immunohistochemistry was employed to assess the expression of tumor necrosis factor (TNF)-α and cyclooxygenase-2 (COX-2). The difference in the severity of inflammation and COX-2 expression was examined using one-way analysis of variance. The correlation of COX-2 expression with the severity of inflammation was analyzed using Spearman's rank correlation test., Result: Until week 13, chronic inflammation and non-hyperplastic and hyperplastic aberrant crypt foci occurred. The severity of inflammation gradually shifted from high moderate to low moderate. TNF-α expression was high in all groups; however, COX-2 expression was gradually lower with longer duration of induction, which corresponded with the severity of inflammation., Conclusion: DMH induction until week 13 without a latency period caused chronic inflammation without the formation of adenoma or adenocarcinoma. A very strong correlation was established between COX-2 expression and inflammation.

Introduction: Inherited retinal dystrophy (IRD) is a group of untreated genetic ocular diseases that mostly affect young people. The number of patients with IRD worldwide, including in developing countries, is growing each year. This literature review aimed to investigate the current utilised genetic screening of IRD worldwide and to propose the most feasible genetics test and diagnostic method for IRD in developing countries, especially Indonesia., Materials and Methods: A literature search was performed in PubMed and Google Scholar databases. Papers conducting wide genome sequencing, including panel sequencing (panel-seq), microarray, whole exome sequencing (WES), whole genome sequencing (WGS) and Sanger sequencing on patients with IRD, were included. Papers were sorted into several groups to visualise the sequencing technology's detection rate. Detection rate comparison analysis was done using the meta-regress protocol in the R program. Whereas the number of novel mutations in each testing tool each year was pooled and compared in the graph., Results: After conducting the literature study, 37 papers were sorted from 451 results. Most studies conducted a panel-seq with 16 records followed by WES with seven records. The detection rate of the WES meta-analysis was 0.66, which was slightly better than the panel-seq with 0.55. The number of novel mutation discoveries fluctuated each year with panel-seq as the most prominent finder. Cost factors and the limitation of sequencing devices make panel-seq a more appropriate tool in Indonesia., Conclusion: The most effective selection for evaluated genetic testing was WES. Therefore, panel-seq is more suitable for first-tier genetic testing in Indonesia.

Objective: The aim of this study is to examine the M1 and M2 macrophages distribution in the rat's colon of DMH-induced inflammation associated colorectal cancer., Methods: Colon tissue of three groups of 4 rats that induced using 1,2 dimethylhydrazine (DMH) at 30 mg/kg bw every week for 9, 11, and 13 weeks were used. The M1 and M2 distribution was examined by using antibody anti iNOS for M1 and anti-CD163 for M2 with immunohistochemistry method. The data was presents in figure and table in the form of percentage., Result: M1 macrophage was found in all groups in the low distribution level (25% - 50%), while M2 macrophage was observed in all groups with 100% distribution. In the longer period of DMH induction, M2 macrophages was distributed more abundant., Conclusion: All of the rat's colon showing chronic inflammation that led to the tumorigenesis.

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.

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 process., 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 /CV 2 < 0.5., Competing Interests: There are no conflicts of interest., (Copyright: © 2022 Journal of Microscopy and Ultrastructure.)

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., Competing Interests: The authors have declared that no competing interests exist.

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.

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. , .

The aim of this study was analyzing the BCR-ABL transcript types of patients with chronic myeloid leukemia (CML) in Dr Sardjito General Hospital, Yogyakarta, Indonesia. This study is very relevant because the data concerning BCR-ABL gene transcript types is very limited in Indonesia. Furthermore, it is important for patient's management, particularly in defining the tyrosine kinase inhibitors (TKIs) therapy and monitoring after therapy. The introduction of TKIs has become a major advance in the management of patients with CML, especially in the chronic phase (CML-CP), in which most patients are diagnosed., Methods: One hundred eighty five (185) of 370 recruited patients were included in this study (2010-2014). RNA samples were isolated from mononuclear cells of peripheral blood of the subjects taken at primary diagnosis. Detection of BCR-ABL gene transcript types was done using multiplex reverse transcriptase PCR (multiplex RT-PCR) and/or nested PCR following the cDNA synthesis. When the first PCR set failed to amplify the BCR-ABL gene, RT-conventional PCR and/or nested PCR would be applied. The proportion of each transcript type was calculated among the BCR-ABL positive CML patients., Results: Approximately 99% (183/185) of CML patients are BCR-ABL positive, with the most common type is major b3a2 (136/183; 74.3%), followed by major b2a2 (41/183; 22.4%). Two samples (1.1%) showed co-expression of b3a2 and b2a2; 1 sample showed co-expression of b3a2 and fragment at 500bp; and 3 samples showed uncommon fragments., Conclusion: Ninety nine percent (99%) of CML patients in Yogyakarta, Indonesia are BCR-ABL positive, with 74.3% have b3a2 transcript, 22.4% have b2a2 trascript, 1.1% have co-expression of b3a2 and b2a2 transcript, and the rest (2.2%) have uncommon bands that still need to be confirmed.

Background: Vascular endothelial growth factor-A (VEGF-A) has been observed as the predominant angiogenic factor in colorectal cancer (CRC) and the assessment of microvessel density (MVD) has been used to quantify tumor neoangiogenesis. This study aimed to determine clinicopathological and prognostic significance of both angiogenic markers in the local CRC patients., Methods: We analyzed tissue samples obtained from 81 cases with CRC. VEGF-A expression and MVD counts were immunohistochemically detected using anti VEGF-A and CD31. The assessments of both markers were classified as low and high. Correlation between VEGF-A expression and MVD value and clinicopathological characteristics were examined using Chi-square test. The overall survival (OS) was plotted using the Kaplan-Meier method., Results: High VEGF-A expression was found more frequently in the rectal location (P=0.042) and T4 tumors (P=0.041) compared to their counterparts. Older patients tended to show a higher MVD value compared to younger cases (P=0.062). In addition, survival analysis showed that males had a worse OS compared to females (P=0.029), and VEGF-A expression and MVD count did not correlate with patients' survival., Conclusions: There were significant differences of VEGF-A expression according to tumor location and T invasion. Sex, but not angiogenic markers, had an influence on the survival of CRC patients., Competing Interests: Conflicts of Interest: The authors have no conflicts of interest to declare.

Objective: This study aimed to identify micro-satellite instability (MSI) based on the expression of MMRp (MSH2 and MSH6) and to evaluate the association of MSI and with clinicopathological features in patients with colorectal cancer (CRC). Methods: MMRp expression in 80 tissue samples from patients with adenocarcinoma CRC were evaluated by using anti-MSH2 and -MSH6 antibodies. Loss of MSH2 and/or MSH6 expression was stated as MSI. The association between MSI status and clinicopathological features were analyzed by using binary logistic regression (p<0.05). Results: The frequency of MSI in patients with CRC varied, corresponding to 8.3% (6/72) MSH2 MSI, 36.1% (26/72) MSH6 MSI and 6.9% (5/72) MSH2-MSH6 MSI. Male patients (OR=1.98), with tumor located in colon (OR=1.47) and late stage tumor (OR=1.48) have a tendency of having MSH2 MSI. Male patients (OR=1.4), with tumor located in colon (OR=2.53) and poor tumor differentiation (OR=3.02) have a tendency to encounter MSH6 MSI. Male patients (OR=4.93) with late stage tumor (OR=1.69) have a tendency of having MSH2-MSH6 MSI. Conclusion: Patients more likely to have MSH2 MSI are males, and/or having tumor located in colon, and /or having late stage tumor. Patients more likely to have MSH6 MSI are males, and/or having tumor located in colon, and/or having tumor with poor differentiation. Patients who have greater tendency to have MSH2 and MSH6 MSI are males, and/or having late stage tumor., (Creative Commons Attribution License)

Epstein-Barr (EBV) infection and presence of a nasopharyngeal cancer (NPC) case in the family increases the risk of developing NPC. Aberrant anti-EBV immunoglobulin A (IgA) antibodies (EBV-IgA) may be present in the sera of non-cancer individuals and predict NPC. Limited studies report the presence of EBV-IgA antibodies within non-cancer individuals in Indonesia where the disease is prevalent. This study aimed at exploring whether EBV-IgA was found more frequently among first degree relatives of NPC patients and individuals presenting with chronic symptoms in the head and neck area compared to healthy controls. A total of 967 non-cancer subjects were recruited, including 509 family members of NPC cases, 196 individuals having chronic complaints in the head and neck region, and 262 healthy donors of the local blood bank. Sera were analyzed using a standardized peptide-based EBV-IgA ELISA. Overall, 61.6% of all individuals had anti-EBV IgA reactivity equal to or below cut off value (CoV). Seroreactivity above CoV was significantly higher in females (38.7%) compared to males (28.7%) (p = 0.001). Older individuals had more seroreactivity above CoV (42.5%) than the younger ones (26.4%) (p< 0.001). Seroprevalence was significantly higher in family members of NPC patients (41.7%), compared to 32.7% of individuals with chronic head and neck problems (p = 0.028) and 16.4% healthy blood donors (p< 0.001). As conclusion, this study showed a significant higher seroprevalence in healthy family members of NPC cases and subjects presenting with chronic symptoms in the head and neck area compared to healthy individuals from the general community. This finding indicates that both groups have elevated risk of developing NPC and may serve as targets for a regional NPC screening program.

Background: Polymorphic bases in several exons of the BCR gene have been found in several studies of the BCR-ABL fusion gene . Most of the polymorphisms do not have any implications for the primary structure of the BCR-ABL protein. Nucleotide changes are often located in the area close to the fusion region, and therefore may influence primer annealing. Our previous work failed to amplify 15 of 200 samples from BCR-ABL positive chronic myelogenous leukemia (CML) patients using multiplex PCR, the standard method to detect BCR-ABL transcripts used in our institution. The failure was considered due to problems in primer annealing caused by sequence variations. Sequence analysis of BCR-ABL fusion gene breakpoint types in CML patients has never been hitherto performed in Indonesia. Therefore, the aim of this study was to perform sequence analysis of several samples that did not show amplification using the standard method. Methods: Fifteen samples were qualitatively amplified by two-step PCR using inner primers in the 2nd PCR to determine the breakpoint type of the BCR-ABL fusion gene. The 2nd PCR products were used as templates to perform sequence analysis, and the results were compared to those in genbank. Result: Seven and 5 of 15 samples were confirmed as major b3a2 and major b2a2, respectively. One sample featured a combination of b3a2 and b2a2, and 2 samples a combination of b3a2 and b2a2 with an additional fragment at 500bp. Sequence analysis showed 3 sequence variations in the major b3a2 breakpoint. One had been reported earlier (c.3296T>C) but the others (c.3245C>T and c.3359T>C) were novel. Fragments at 500bp were confirmed as b3a2 and similar sequence b3a2 in genbank. Conclusion: This study found two new genetic variations in the BCR gene in BCR-ABL fusion cases., (Creative Commons Attribution License)

Single nucleotide polymorphism (SNP) detection has been used extensively for genetic association studies of diseases including cancer. For mass, yet accurate and more economic SNP detection we have optimized tetra primer amplification refractory mutation system polymerase chain reaction (ARMS PCR) to detect three SNPs in the cytochrome P450 2E1 (CYP2E1) gene locus; i.e. rs3813865, rs2070672 and rs3813867. The optimization system strategies used were (1) designing inner and outer primers; (2) determining of their optimum primer concentration ratios; and (3) determining of the optimum PCR annealing temperature. The tetra primer ARMS PCR result could be directly observed using agarose gel electrophoresis. The method succesfully determined three SNPs in CYP2E1 locus, the results being consistent with validation using DNA sequencing and restriction fragment length polymorphisms (RFLP).