Soluble Human Leukocyte Antigen Molecules Detected in Orofacial Cleft Patients: A Case-Control Study

Abstract Objective: To compare soluble HLA-C and HLA-DR molecules present in the plasma of orofacial cleft and non-orofacial cleft populations. Material and Methods: Orofacial cleft patients were recruited using an accidental sampling approach (n=15). Peripheral blood was collected from the participants and processed for Enzyme Linked Immunosorbent Assay (ELISA) against HLA-C and HLA-DR with specific antibodies. The absorbance was calculated utilizing ELISA reader. Data were statistically analyzed using an independent t-test to compare the disease and control groups. Results: The levels of soluble HLA-C and HLA-DR were significantly higher in the diseased group compared to the control group (p<0.05). Conclusion: The role of HLA molecules in non-communicable disease and congenital anomalies, particularly orofacial cleft, remains speculative despite the positive results of this study and those of previous investigations. It suggests that the variables examined may affect specific pathways involved in the pathogenesis of orofacial cleft, and predispose the individuals concerned to the oral cleft.

High Mobility Group Box 1 and Heat Shock Protein-70 Expression Post (-)-Epigallocatechin-3-Gallate in East Java Green Tea Methanolic Extract Administration During Orthodontic Tooth Movement in Wistar Rats

Abstract Objective: To investigate the expression of High Mobility Group Box 1 (HMGB1) and Heat Shock Protein-70 (HSP-70) during orthodontic tooth movement (OTM) after (-)- Epigallocatechin-3-Gallate (EGCG) in East Java Green Tea (Camelia Sinensis) Methanolic Extract (GTME) administration in vivo. Material and Methods: 28 Wistar rats (Rattus Novergicus) was used and divided into 4 groups accordingly: K- without EGCG and OTM; K+ with OTM, without EGCG for 14 days; T1with OTM for 14 days and EGCG for 7 days; treatment group 2 (T2) with OTM and EGCG for 14 days. OTM animal model was achieved through the installation of the OTM device by means of NiTi close coil spring with 10g force placed between the first incisor and first maxillary molars. The samples were terminated on Day 14. The pre-maxillary was isolated for the immunohistochemical examination. Analysis of Variance (ANOVA) then continued with Tukey Honest Significant Difference (HSD) (p<0.05) was performed to analyze the data. Results: The highest HMGB1 and HSP-70 expression were found in the K+ group pressure side, meanwhile the lowest HMGB1 and HSP-70 expression were found in K- group tension side in the alveolar bone. There was a significant decrease of HMGB1 and HSP-70 expression in T2 compared to T1 and K+ with significant between groups (p<0.05; p=0.0001). Conclusion: The decreased expression of HMGB1 and HSP-70 in alveolar bone of OTM wistar rats due to post administration of GTME that consisted EGCG.

The potency of chitosan-based Pinus merkusii bark extract nanoparticles as anti-cancer on HeLa cell lines.

BACKGROUND AND AIM: Cervical cancer accounts for the fourth as a cause of death from cancer in women worldwide, with more than 85% of events and deaths occurring in developing countries. The main problems of chemotherapy are the lack of selectivity and drug resistance. This study aimed to investigate the signal transduction of chitosan-based Pinus merkusii bark extract nanoparticles (Nano-PMBE) as an anticancer on HeLa cell line. MATERIALS AND METHODS: Nano-PMBE was prepared based on the ionic gelation method. Its anticancer activities in HeLa cells were investigated through cytotoxicity test, cell cycle, and apoptosis analysis. The expression of p53 and caspase-9 was also observed. RESULTS: The results showed that Nano-PMBE has a size of 394.3 nm. Meanwhile, the Nano-PMBE was cytotoxic to HeLa cells (IC50 of 384.10 µg/ml), caused G0/G1 phase arrest and cell apoptosis in HeLa cells. Besides, the expression of p53 and caspase-9 has increased. CONCLUSION: The results showed a notable anticancer effect of Nano-PMBE by arresting the cell cycle and inducing apoptosis in HeLa cells, suggesting that it might have therapeutic potential for cervical cancer. Further research is needed to find out more about the anticancer mechanism of Nano-PMBE in HeLa cells to in vivo and clinical studies.

The Densitometric Analysis of Protein Pattern in Cleft Lip and Palate Patients.

OBJECTIVES: Cleft lip and palate (CLP) belongs to the congenital anomaly that is clinically seen as cleft in lip, alveolar bone, palate, and nasal septum. The patients suffer from esthetic and various functional defects. CLP is resulted from impaired palatogenesis during the embryonic phase. The etiology of CLP is influenced by genetic, environmental, and combination of both. According to the literature, CLP is highly associated with defect in interferon regulatory factor 6 (IRF6) and poliovirus receptor-like (PVRL1) genes. The present study aimed to investigate the total protein profile and to identify protein IRF6 and PVRL1 in plasma of CLP patients. MATERIALS AND METHODS: Dot-Blot analysis was performed to identify protein target of IRF6 and PVRL1. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis was performed in gel concentration 12% using plasma of CLP patients, their parents, and control population. The gels were stained by Coomassie blue afterward. Gels were analyzed through ImageLab 5.2.1 software. RESULTS: The intensity of major bands in CLP patients was darker than control group, but remains similar to the parents group. The target protein IRF6 and PVRL1 were positively identified through Dot-Blot. Retardation factor value was significantly different in major bands of CLP patients compared to control group. CONCLUSION: There pattern of protein profile in CLP patients was different compared to non-CLP.