Interleukin 10 Induces the Expression of Membrane-Bound HLA- G and the Production of Soluble HLA-G on HeLa CCL-2 Cells.

BACKGROUND: Interleukin-10 is a cytokine that has a pleiotropic effect on the immune system and inflammation. IL-10 can contribute to the anti-tumour immune response by increasing HLA-G expression. AIM: This study aimed to determine the effect of IL-10 induction on membrane HLA-G expression and soluble HLA-G production of HeLa CCL-2 cells. METHODS: HeLa CCL-2 cells were cultured in the well plate and divided into 4 groups consist of 1 control group that was not induced by IL-10 and 3 treatment groups that were induced by IL-10 500 ng/ml, 1000 ng/ml and 2000 ng/ml respectively. All groups were incubated for 48 hours in a 37°C incubator at 5% CO2 atmospheric pressure. HLA-G measurements were carried out both in cell lysate and cell culture supernatant using ELISA and in membrane-bound using immunofluorescence method. The expression of HLA-G in membrane-bound calculated using the ImageJ application. Data obtained were analysed by ANOVA and LSD test. RESULTS: In the control group, the HLA-G level in the culture supernatant was higher than in cell lysate (p = 0.000), as well as in all treatment groups (p = 0.000). There were significant differences between the treatment group (p = 0.000) and within the treatment group (p = 0.000) at HLA levels. The highest expression of HLA-G in HeLa cell membranes found in cell culture induced by IL-10 concentrations of 500 ng/ml, i.e., 59.28 AU in view. HLA-G membrane expression in the IL-10 1000 ng/ml induced group was significantly different from all treatment groups (p = 0.000). CONCLUSION: HeLa CCL-2 cells express HLA-G on the membrane and release dissolved HLA-G without induction of IL-10 although IL-10 induction augments the presence and the production of HLA-G in HeLa CCl-2 cells.

ANTIBACTERIAL ACTIVITY OF DRACONTOMELON DAO EXTRACTS ON METHICILLIN-RESISTANT S. AUREUS (MRSA) AND E. COLI MULTIPLE DRUG RESISTANCE (MDR).

BACKGROUND: Staphylococcus aureus, methicillin-resistant and Escherichia coli, multidrug-resistant included in the list of antibiotic-resistant priority pathogens from WHO. As multidrug-resistant bacteria problem is increasing, it is necessary to probe new sources for identifying antimicrobial compounds. Medicinal plants represent a rich source of antimicrobial agents. One of the potential plants for further examined as antibacterial is Dracontomelon dao (Blanco) Merr. & Rolfe. The present study designed to find the antibacterial activity of D. dao stem bark extracts on Methicillin-resistant S. aureus (MRSA) and E. coli Multiple Drug Resistance (MDR), followed by determined secondary metabolites with antibacterial activity and determined the value of MIC (minimum inhibitory concentration) and MBC (minimum bactericidal concentration). MATERIALS AND METHODS: D. dao stem bark extracted using 60% ethanol. Disc diffusion test methods used to find the antibacterial activity, following by microdilution methods to find the value of MIC and MBC. Secondary metabolites with antibacterial activity determined by bioautography using TLC (thin layer chromatography) methods. RESULTS: D. dao stem bark extracts are sensitive to MSSA, MRSA and E.coli MDR bacteria. The inhibition zone is 16.0 mm in MSSA, 11.7 mm in MRSA and 10.7 mm in E. coli MDR. The entire MBC/MIC ratios for MSSA, MRSA and E.coli MDR is lower than 4. The ratio showed bactericidal effects of D. dao stem bark extracts. In TLC results, colorless bands found to be secondary metabolites with antibacterial activity. CONCLUSION: D. dao stem bark extracts are potential to develop as antibacterial agent especially against MRSA and E. coli MDR strain.

Caloric restriction improves glucose homeostasis, yet increases cardiometabolic risk in caveolin-1-deficient mice.

BACKGROUND AND PURPOSE: The plasma membrane protein caveolin-1 (CAV-1) has been shown to be involved in modulating glucose homeostasis and the actions of the renin-angiotensin-aldosterone system (RAAS). Caloric restriction (CR) is widely accepted as an effective therapeutic approach to improve insulin sensitivity and reduce the severity of diabetes. Recent data indicate that polymorphisms of the CAV-1 gene are strongly associated with insulin resistance, hypertension and metabolic abnormalities in non-obese individuals. Therefore, we sought to determine whether CR improves the metabolic and cardiovascular (CV) risk factors in the lean CAV-1 KO mice. MATERIALS/METHODS: Twelve- to fourteen-week-old CAV-1 knockout (KO) and genetically matched wild-type (WT) male mice were randomized by genotype to one of two dietary regimens: ad libitum (ad lib) food intake or 40% CR for 4 weeks. Three weeks following the onset of dietary restriction, all groups were assessed for insulin sensitivity. At the end of the study, all groups were assessed for fasting glucose, insulin, HOMA-IR, lipids, corticosterone levels and blood pressure (BP). Aldosterone secretion was determined from acutely isolated Zona Glomerulosa cells. RESULTS: We confirmed that the CAV-1 KO mice on the ad lib diet display a phenotype consistent with the cardiometabolic syndrome, as shown by higher systolic BP (SBP), plasma glucose, HOMA-IR and aldosterone levels despite lower body weight compared with WT mice on the ad lib diet. CAV-1 KO mice maintained their body weight on the ad lib diet, but had substantially greater weight loss with CR, as compared to caloric restricted WT mice. CR-mediated changes in weight were associated with dramatic improvements in glucose and insulin tolerance in both genotypes. These responses to CR, however, were more robust in CAV-1KO vs. WT mice and were accompanied by reductions in plasma glucose, insulin and HOMA-IR in CAV-1KO but not WT mice. Surprisingly, in the CAV-1 KO, but not in WT mice, CR was associated with increased SBP and aldosterone levels, suggesting that in CAV-1 KO mice CR induced an increase in some CV risk factors. CONCLUSIONS: CR improved the metabolic phenotype in CAV-1 KO mice by increasing insulin sensitivity; nevertheless, this intervention also increased CV risk by inappropriate adaptive responses in the RAAS and BP.

Association of thrombocytopenia with splenomegaly in malaria patients in East Kalimantan: A cross-sectional, retrospective study.

Background: Malaria still presents as a major health problem in Indonesia and specifically in East Kalimantan. One common sign found in malaria patient is thrombocytopenia, the mechanism of which is still unclear. Several studies have suggested some mechanisms, one of which is splenomegaly. This study aimed to discover the association between thrombocytopenia and splenomegaly of malaria patients in East Kalimantan. Methods: This study was a descriptive retrospective study with clinical and laboratory data obtained from the medical records of malaria patients in four major public hospitals from January 2015 to July 2018. The association between thrombocytopenia with splenomegaly was analysed using Chi-Square test. Results: A total of 215 patients were included; 189 male (87.9%) and 26 female (12.1%). The etiologic agents found in these patients were  Plasmodium vivax (43.2%),  Plasmodium falciparum (42.8%), and mixed infection ( Plasmodium falciparum and  Plasmodium vivax) (4.6%). The thrombocyte count was normal in 28 patients (13%) and decreased in 187 patients (87%). Among patients with thrombocytopenia, the percentage of mild, moderate and severe thrombocytopenia was 18.2%, 43.8% and 33%, respectively. Splenomegaly was found in only 11 patients (5.1%). We found no association between thrombocytopenia with splenomegaly (p=0.61). Conclusions: We conclude that thrombocytopenia is not associated with splenomegaly in these malaria patients.