Endometrial receptivity defects MUC-1 and COX-2 polymorphisms in endometriosis.

The endometrium produces MUCIN-1 (MUC-1) and cyclooxygenase-2 (COX-2), which are essential for implantation. MUC-1 is required for adhesion, while COX-2 is necessary for decidualization. Variations or polymorphisms in MUC-1 and COX-2 can lead to changes in endometrial receptivity. This study investigated the relationship between MUC-1 and COX-2 polymorphisms and endometrial receptivity in endometriosis patients. Blood DNA samples were collected from 35 patients with endometriosis and 32 healthy patients between days 19 to 24 of their menstrual cycle (secretory phase). MUC-1 polymorphism was determined using the Amplification Refractory Mutation System (ARMS), and COX-2 gene polymorphism was assessed using Polymerase Chain Reaction-Restriction Fragment Length Polymorphism (PCR-RFLP). The frequency distribution of gene polymorphisms between the two groups was compared using bivariate analysis. There were seven genotypic combinations of MUC-1 and COX-2: AAGC; AAGG; GACC; GAGC; GAGG; GGGC; GGGG. The AAGC genotype combination test was significant, with an OR=6.43 (95% CI:1.09-7.62) and p=0.01. In conclusion, combining MUC-1 and COX-2 (AAGC) genotypes results in endometrial receptivity defects in endometriosis.

Mutational heterogeneity between different regional tumour grades of clear cell renal cell carcinoma.

Only a limited number of studies have explored the possible associations between tumour grade and mutated genes in clear cell renal cell carcinoma (ccRCC), and we set out to investigate this further using a multiple sampling and next generation sequencing (NGS) approach in a series of ccRCCs. Multiple regions were sampled from formalin-fixated paraffin-embedded ccRCC tumour blocks from seven patients. In 27 samples from six patients, we performed targeted NGS using a custom 42-gene panel based on the most frequently mutated genes in ccRCC reported in public databases. In four samples from the seventh patient, we performed whole exome sequencing (WES) and array comparative genomic hybridisation for detection of copy number variants (CNVs). Mutated genes and the tumour grades of the samples in which they had been identified were compared both within and between all individual tumours. CNVs were compared across all samples from patient 7. We identified clear genetic heterogeneity within and across tumours, but VHL mutations were seen in all patients. Looking across all samples, we identified eleven genes that were only mutated in samples with one particular tumour grade. However, these genes were never mutated in all samples with that tumour grade. Increasing chromosomal instability corresponded with increasing tumour grade, but we observed minimal association between tumour grade and total mutational load in the WES data. Our study confirms the genetic heterogeneity and tumour grade heterogeneity of ccRCC. Although a relatively small number of samples was analysed, genes were identified that could potentially be specific, though insensitive, markers of higher ccRCC tumour grades.

Prognostic Value of Lymphangiogenesis Determinants in Luminal and Non-luminal Breast Carcinomas

Background: Breast carcinomas (BCs) are sub-classified according to the molecular characteristics into luminal and non-luminal subtypes that clinically show different biological behavior, treatment and prognosis. BCs spread primarily through lymphatic vessels using cascade processes of lymphagiogenesis in which VEGF-C plays an important role during lymph node metastasis. Prognostic value of VEGF-C in luminal and non-luminal BC is still unclear and has not been studied thoroughly to clarify and define prognosis and therapeutic monitoring. Aim: To define the prognostic value of lymphangiogenesis on survival rates of luminal and non-luminal subtypes BC. Materials and Methods: This study applied prospective cohort design, using 130 patients of invasive duct carcinoma of the breast, stage I-IIIA, from Sardjito General Hospital, Indonesia and subsequent longitudinal follow-up. Immunohistochemical staining was carried out using anti-ER, -PR, -Her-2, VEGF-C, VEGFR-3 and D2-40 antibodies. The related clinicopathologic characteristics of BC patients and lymphangiogenesis determinants, including VEGF-C expression, were statistically analyzed. Results: In non-luminal BC subtypes, VEGF-C expression (HR=0.04; 95% CI=0.01-0.41), lymph node metastasis (HR=0.14; 95% CI=0.04-0.55) and stage (HR=0.30; 95% CI= 0.02-0.76) were determined as independent prognostic factors on survival rates. However, the lymphangiogenesis determinants were not associated with the survival rates of luminal BC subtypes. Conclusion: This study suggested that lymphangiogenesis affects survival rates of non-Luminal subtype rather than the luminal subtypes of BC.

Prognostic Value of Clinicopathological Factors for Indonesian Breast Carcinomas of Different Molecular Subtypes

Background: Breast carcinoma (BC) is a heterogeneous disease due to its different molecular profiles i.e. luminal (luminal A and luminal B) and non-luminal (HER2 positive and triple negative) subtypes. Prognostic value of clinicopathological factors of Indonesian BC of different molecular subtypes has never been reported previously. This study aims to elaborate prognostic impacts on Indonesian BCs focusing on separate molecular subtypes. Methods: A hundred and fifty cases of invasive BC, stage I-IIIA, in Sardjito Hospital, Indonesia, were stained using anti ER, PR, HER2 and Ki-67 antibodies. Survival and prognostic values were statistically analyzed. Results: Compared to the luminal subtypes, the non-luminal subtypes demonstrated higher proportions of intermediate-to-high grade, stage IIIA, positive lymph node infiltration and mortality. The triple negative subtype was typically intermediate-to-high grade, stage IIIA and with a high relative death risk. Luminal A lesions were characteristically low grade, stage I-II and less likely to cause death. Conclusion: In non-luminal BC, staging and lymph node metastasis are independent prognostic factors for survival in HER2 positive and triple negative subtypes, respectively. In luminal BC, clinicopathological factors demonstrated no influence on survival. This study suggests that staging and lymph node metastasis are correlated with survival in non-luminal Indonesian BCs.

Nestin expressions of exposed pulp after direct pulp capping by calcium hydroxide and platelet rich plasma.

OBJECTIVE: The aim of this study was to evaluate nestin expression of pulp tissue following direct pulp capping with platelet-rich plasma (PRP). MATERIALS AND METHODS: The thirty sound teeth from Sprague-Dawley rats were used and divided into two groups: Groups 1, teeth were capped with calcium hydroxide/Ca(OH)2 (n = 15) and Group 2 with PRP (n = 15). After 1(st), 7(th), and 21(st) days, respectively, 5 teeth each group (American Dental Association 41) were processed for light microscopic examination. Expressions of nestin were assessed by immunohistochemical techniques. RESULTS: Nestin expression of Ca(OH)2 on the distance place of exposure at 1(st) and 7(th) days were 80% and at 21(st) day were 60%. Nestin expression of PRP on the distance place at 1(st) day was 80%, 7(th) 100%, and 21(st) day was 80%. At day 21 observation, Kruskal-Wallis test shows nestin expression was increased significantly in PRP groups (P < 0.05), but it was not increase significantly compare with Ca(OH)2. CONCLUSION: PRP had ability as a direct pulp capping material to induce nestin expression.

Clinicopathological features of indonesian breast cancers with different molecular subtypes.

BACKGROUND: Breast cancer is a heterogeneous disease with molecular subtypes that have biological distinctness and different behavior. They are classified into luminal A, luminal B, Her-2 and triple negative/basal-like molecular subtypes. Most of breast cancers reported in Indonesia are already large size, with high grade or late stage but the clinicopathological features of different molecular subtypes are still unclear. They need to be better clarified to determine proper treatment and prognosis. AIM: To elaborate the clinicopathological features of molecular subtypes of breast cancers in Indonesian women. MATERIALS AND METHODS: A retrospective cross-sectional study of 84 paraffin-embedded tissues of breast cancer samples from Dr. Sardjito General Hospital in Central Java, Indonesia was performed. Expression of ER, PR, Her-2 and Ki-67 was analyzed to classify molecular subtypes of breast cancer by immunohistochemistry. The relation of clinicopathological features of breast cancers with molecular subtypes of luminal A, luminal B, Her-2 and triple negative/basal-like were analyzed using Pearson's Chi-Square test. A p-value of <0.05 was considered statistically significant. RESULTS: Case frequency of luminal A, Luminal B, Her-2+ and triple negative/basal-like subtypes were 38.1%, 16.7%, 20.2% and 25%, respectively. Significant difference was found in breast cancer molecular subtypes in regard to age, histological grade, lymph node status and staging. However it showed insignificant result in regard to tumor size. Luminal A subtype of breast cancer was commonly found in >50 years old women (p:0.028), low grade cancer (p:0.09), negative lymph node metastasis (p:0.034) and stage III (p:0.017). Eventhough the difference was insignificant, luminal A subtype breast cancer was mostly found in small size breast cancer (p:0.129). Her-2+ subtype breast cancer was more commonly diagnosed with large size, positive lymph node metastasis and poor grade. Triple negative/basal-like cancer was mostly diagnosed among <50 years old women. CONCLUSIONS: This study suggests that immunohistochemistry-based subtyping is essential to classify breast carcinoma into subtypes that vary in clinicopathological features, implying different therapeutic options and prognosis for each subtype.

Hepatitis E virus infection in two different regions of Indonesia with identification of swine HEV genotype 3.

Hepatitis E is an emerging disease with a high incidence globally. Few data are available on hepatitis E virus (HEV) infection in Indonesia. To obtain molecular information on HEV infection in two regions of Indonesia with different customs and swine breeding conditions, serum samples from 137 swine farm workers, 100 blood donors and 100 swine (27 fecal samples also obtained) in Yogyakarta (Central Java) and from 12 and 64 swine farm workers, 42 and 135 local residents and 89 and 119 swine in Tulungagung (East Java) and Mengwi (Bali), respectively, from our previous study, were compared.Serological tests for anti­HEV antibodies by ELISA, HEV­RNA detection by RT­PCR and phylogenetic analysis were performed. The total prevalence of anti­HEV antibodies in humans was higher in Bali(11.6%) than in Java (5.1%; P=0.015). No significant differences in anti­HEV prevalence among swine farm workers and local residents in Java were found. The finding of swine HEV genotype 3 in specimens from Yogyakarta and genotype 4 from Tulungagung and Bali is somewhat different from other reports.We suggest other factors in addition to close contact with swine might play an important role in HEV transmission of non­endemic/related custom groups. To the best of our knowledge, this is the first report on swine HEV genotype 3 in Indonesia.

Clinicopathological significance of lymphangiogenesis and tumor lymphovascular invasion in indonesian breast cancers.

BACKGROUND: Lymphangiogenesis, assessed as lymphovascular density (LVD), is the initial step of generalized tumor lymphovascular invasion (LVI). It also involves VEGF-C as the most important protein family. Lymphangiogenesis among breast cancer cases correlations with several clinicopathological factors are important to determine prognosis and treatment strategies, but results have been controversial and require clarification. AIM: To define correlations between VEGF-C expression, LVD and LVI with several clinicopathological parameters from Indonesian breast cancer patients. MATERIALS AND METHODS: Using a cross-sectional study, a total of 48 paraffin-embedded tissues of breast cancer from Dr. Sardjito General Hospital Indonesia were assessed for VEGF-C expression, LVD and LVI by immunohistochemistry. Correlations of these markers with clinicopathological parameters like patient age, tumor size, lymph node status, grade, ER/PR and Her-2 status, cell proliferation and p-53 expression were investigated by linear analysis. Correlations of VEGF-C expression and LVI with several clinicopathological parameters were analyzed with Coefficient Contingency Chi-Square test. RESULTS: The mean of patients age was 53.0 year, pre and post-menopausal patients accounting for 56.3% and 43.8%, respectively. Some 10.4% were well, 41.7% moderate and 47.9% poorly differentiated. ER positivity was evident in 50% while PR and Her-2 positivity was found in 31.3% and 33.3%, respectively. Breast cancer cells with over-expression of p-53 was 64.6% and with high cell proliferation was 56.3%. Lymph node metastasis was noted in 63.5%, and LVI in 72.9%. Significant correlations were found between LVD and tumor size (p:0.037), grade (p:0.000), lymphnode status (p:0.036), LVI (p:0.003), as well as with p-53 and cell proliferation. There were also significant correlation of VEGF-C (p:0.011) and LVI (p:0.001) with tumor grade. Only ER status was found to have a correlation with tumor size (p:0.027). CONCLUSIONS: This study suggested that in Indonesian breast cancer patients, lymphangiogenesis is correlated with tumor size, grade, lymph node status and tumor lymphovascular invasion, the latter also being related with p-53 over expression and high cell proliferation.

Effect of nifedipine on the expression of bcl-2 protein in rat gingiva.

Bcl-2 is a family of proteins involved in protecting the cell against death stimuli or in promoting cell death. The aim of the present study was to determine the effect of nifedipine treatment on the expression of bcl-2 protein in rat gingival tissues. Rats were given gastric intubation with various concentrations and durations of nifedipine. Nifedipine-untreated and dimethylsulfoxide (DMSO)-treated animals served as control groups. The gingival tissues were dissected and the expression of bcl-2 protein was determined immunohistochemically. The results showed that the numbers of bcl-2-positive cells in the gingiva of nifedipine-treated animals were significantly higher than in the control groups. These numbers increased parallel to increased concentration and duration of nifedipine treatment. The results suggest that nifedipine treatment may induce the expression of bcl-2 protein in rat gingival tissue in a dose- and duration-dependent fashion and that this proto-oncogenic protein may play a role in nifedipine-induced gingival hyperplasia.