Coelomic Fluid of Lumbricus rubellus Synergistically Enhances Cytotoxic Effect of 5-Fluorouracil through Modulation of Focal Adhesion Kinase and p21 in HT-29 Cancer Cell Line.

Coelomic fluid of Lumbricus rubellus (CFL) has attracted interest due to its pharmacological properties, including antitumor effect. Furthermore, it is necessary to evaluate the response to treatment with new cancer therapeutic agents. This study aims to investigate whether the combination of CFL and 5-fluorouracil could reduce FAK protein level and iCa2+ and enhance p21 level. Furthermore, it is necessary to evaluate the response to treatment with new cancer therapeutic agents. After 24 hours of treatment, it was necessary to assess the percentage of apoptosis, FAK, and p21 protein expression by flow cytometry. iCa2+ concentration was measured using immunofluorescence. The combination therapy of CFL with 5-fluorouracil potently suppressed six treatment groups were included in this study. HT-29 cell lines were cultured and divided into six groups: group 1 was treated with vehicle (negative control), groups 2-5 were treated with 5-fluorouracil, groups 3-5 were treated with either CFL 5, 10, or 20 µg/ml immediately after 5-fluorouracil, and group 6 was treated with CFL 20 µg/ml, the progression of colorectal cancer. Combination of CFL and 5-fluorouracil significantly decreased FAK expression (p<0.05), iCa2+ (p<0.05), and increased p21 expression (p<0.05) in HT-29 cells. Our results suggest that CFL has an anticancer potential in colorectal cancer when combined with 5-fluorouracil.

Vitamin D3 induces stem cell activation via Lgr5-Bmi1 expression and improving mouse colitis histology index.

Conventional therapy for inflammatory bowel disease using long-term anti-inflammatory drugs does not seem to provide optimal results. Adjuvant therapy using vitamin D3 is believed to have an essential role in repairing the colonic mucosa through the activation of colonic stem cells. The aim of this study was to demonstrate the effect of vitamin D3 in mucosal repair through stem cell activation, marked by leucin-rich repeat-containing G protein-coupled receptor 5 (Lgr5) and B lymphoma Mo-MLV insertion region 1 (Bmi1) expression and decrease the mouse colitis histology index (MCHI) score. In this study, 50 Mus musculus strain BALB/c were divided into five groups: negative control group, colitis group, and colitis groups with vitamin D3 administration of 0.2 mcg, 0.4 mcg, and 0.6 mcg per 25 g body weight for seven days. Dextran sulfate sodium (DSS) 5% was used to induce colitis. Lgr5-Bmi1 expression was measured using immunodoublestain fluorescent labeling method. Our data suggested that administration of vitamin D3 significantly increased expression of Lgr5-Bmi1 in the colonic mucosa. The colitis group treated with the highest dose of vitamin D3 (0.6 mcg/25 gram) showed the lowest MCHI score (3.60±0.64) while the lowest dose of vitamin D3 had the highest MCHI score (12.60±1.47). In conclusion, by stimulating stem cells, vitamin D3 administration stimulates mucosal regeneration, as demonstrated by upregulated expression of Lgr5-Bmi-1.

Anxiety, Depression, and Associated Factors among General Population in Indonesia during COVID-19 Pandemic: A Cross-Sectional Survey.

Introduction: The 2019 coronavirus pandemic (COVID-19) has affected the physical and mental health of individuals, families, and communities worldwide including Indonesia. This study aimed to examine anxiety and depression in the general population and factors related to anxiety and depression due to the COVID-19 pandemic. Methods: This study employed an online cross-sectional survey of 1149 respondents. We assessed self-reports regarding current health conditions and exposure to COVID-19, anxiety, and depression in the general population in Indonesia. Results: The results showed that 26.6% and 30.5% of the participants experienced mild to severe anxiety and depression, respectively. The ordinal regression test showed that anxiety in the community was significantly related to age, feeling infected with COVID-19, feeling that a friend/colleague is infected with COVID-19, sufficient information regarding COVID-19, and the types of symptoms that are felt (fever, cough, and cold/sore throat, difficulty breathing). Besides, education level, occupation, feeling that family is infected with COVID-19, symptoms experienced, and anxiety were significantly related to depression. Conclusion: The COVID-19 pandemic has caused anxiety and depression in the general population in Indonesia. This study's results can be a catalyst in providing psychological interventions for the general public facing the COVID-19 pandemic.

Peptide Sequence of Pili Subunit Protein 49.8 kDa Shigella flexneri as Antigenic Epitope for Shigellosis Vaccine Development.

Objectives: This study investigates the amino acid sequence and identifies antigenic epitopes of 49.8 kilodalton (kDa) pili protein Shigella flexneri, which will be used as candidates for the shigellosis vaccine. Materials and Methods: Our study is a prospectively descriptive laboratory. We used bacterial isolate of S. flexneri pili isolation was performed using a pili cutter and sodium dodecyl-sulfate polyacrylamide gel electrophoresis. The amino acid sequences were analyzed using liquid chromatography dual mass spectrometry (LC-MS/MS) method in the proteomic laboratory. The target epitope antigenicity analysis was tested using Kolaskar and Tongaonkar Antigenicity software. The Bepired Linear Epitope Prediction software is used for epitope mapping. PymOL software was used for the visualization of proteins and molecular docking. Peptides and antibodies were applied to hemagglutination test and immune response was tested using the dot blot method. Results: LC-MS/MS analysis results from the mascot server showed that the 49.8 kDa pili protein is S. flexneri similar to the flagellin protein of S. flexneri 1235-66 (ID I6H2T2). The results of antigenicity analysis and epitope mapping showed that areas of protein that has the most potential and antigenic epitopes are the regions 98-111 and 263-290 with the amino acid sequences, QSSTGTNSQSDLDS (Q-S) and DTTITKAETKTVTKNQVVDTPVTTDAAK (D-K). The results of the molecular docking interaction test between the peptide and the B-cell receptor have a low binding energy. Peptide Q-S and peptide D-K antigens are hemagglutinin molecules because they can agglutinate erythrocytes. The immune response between peptide antigens and anti-peptide antibodies can react based on color gradations in the dotblot method. Conclusion: The amino acid sequences Q-S and D-K are potentially antigenic epitopes. These peptides can be used to develop candidates for shigellosis vaccine.

Shigella flexneri vaccine development: Oral administration of peptides derived from the 49.8 kDa pili protein subunit activates the intestinal immune response in mice.

Background and Aim: The morbidity and mortality of Shigella infections remain a global challenge. Epitope-based vaccine development is an emerging strategy to prevent bacterial invasion. This study aimed to identify the ability of the 49.8 kDa pili subunit adhesin protein epitope of Shigella flexneri to induce an intestinal immune response in mice. Materials and Methods: Thirty adult male Balb/c mice were divided into a control group, cholera toxin B subunit (CTB) group, CTB+QSSTGTNSQSDLDS (pep_1) group, CTB+DTTITKAETKTVTKNQVVDTPVTTDAAK (pep_2) group, and CTB+ ATLGATLNRLDFNVNNK (pep_3). We performed immunization by orally administering 50 µg of antigen and 50 µl of adjuvant once a week over 4 weeks. We assessed the cellular immune response by quantifying T helper 2 (Th2) and Th17 using flow cytometry. In addition, we assessed the humoral immune response by quantifying interleukin (IL-4), IL-17, secretory immunoglobulin A (sIgA), and ß-defensin using enzyme-linked immunoassay. Statistical analysis was performed using one-way analysis of variance and Kruskal-Wallis test. Results: Peptide oral immunization increases the cellular immune response as reflected by the increase of Th2 (p=0.019) and Th17 (p=0.004) cell counts, particularly in the CTB_pep_1 group. Humoral immune response activation was demonstrated by increased IL-4 levels, especially in the CTB+pep_3 group (p=0.000). The IL-17 level was increased significantly in the CTB+pep_1 group (p=0.042). The mucosal immune response was demonstrated by the sIgA levels increase in the CTB+pep_3 group (p=0.042) and the ß-defensin protein levels (p=0.000). Conclusion: All selected peptides activated the cellular and humoral immune responses in the intestine of mice. Further studies are necessary to optimize antigen delivery and evaluate whether the neutralizing properties of these peptides allow them to prevent bacterial infection.

Role of vitamin D in Wnt pathway activation for colonic epithelial cell differentiation.

OBJECTIVES: Inflammatory bowel disease (IBD) is a medical condition that represents a pathological form of inflammation, causing damage to the colonic mucosa. Adjunctive vitamin D therapy may activate the Wnt/ß-catenin pathway that results in cell differentiation and proliferation via stem cell signalling. This study aims to evaluate the effect of vitamin D on ß-catenin and cytokeratin 20 (KRT20) as markers of Wnt pathway activation for colonic cell repair. METHODS: For the experiment, we used 30 musculus mice strains of BALB/c, which were categorised into five groups; the control group (K-) and four other groups, where colitis was induced by dextran sulphate sodium (DSS) for seven days. On the seventh day, the remaining three groups were administered vitamin D with an initial dose of 0.2 µg/25.0 g, 0.4 µg/25.0 g and 0.6 µg/25.0 g until day 14. An objective index of disease activity and a histological score were required as markers of inflammation to evaluate the results of the clinical trials. RESULTS: ß-catenin and KRT20 showed a significant increase in the proliferation index of vitamin D at a dose of 0.6 µg/25.0 g (91.50 ± 4.09 and 48.75 ± 2.28, respectively; p < 0.05) compared to the colitis group. CONCLUSIONS: This study demonstrates that vitamin D could be used as an induction agent of Wnt activation for healing colonic mucosa via multipotent stem cells.

Role of Lymphocytes CD4/CD8 Ratio and Immunoglobulin G Cytomegalovirus as Potential Markers for Systemic Lupus Erythematosus Patients with Periodontal Disease.

OBJECTIVES: The aim of the study was to analyze the correlation between periodontitis severity in systemic lupus erythematosus (SLE) with CD4/CD8 lymphocytes ratio and cytomegalovirus gamma immunoglobulin (IgG CMV) level. MATERIALS AND METHODS: This is a descriptive study using a cross-sectional approach that included 93 subjects who were diagnosed with SLE in Rheumatology Department, Saiful Anwar Hospital, during 2017 to 2019. Periodontitis severity was assessed by periodontal Index (PI). CD4/CD8 lymphocyte ratio was determined using flow cytometry and IgG CMV levels using enzyme-linked immunosorbent assay. STATISTICAL ANALYSIS: The differences among the three groups were analyzed using analysis of variance. Correlation among the groups was calculated using Spearman/Pearson correlation coefficient test, while regression analysis was done using Statistical Package for the Social Sciences. RESULTS: The mean of periodontitis severity and standard deviation in SLE was 2.66 ± 1.02. There were negative correlation between CD4/CD8 lymphocyte ratio with periodontal index (r = -0.971) and positive correlation between IgG CMV level with periodontal index (r = 0.977). CONCLUSIONS: Inverted CD4/CD8 ratio and IgG CMV were found associated with periodontitis severity in SLE patient. Further research was recomended that CD4/CD8 lymphocytes ratio and IgG CMV can be used as a potensial marker of periodontitis severity in SLE patients.

Generating mucosal and systemic immune response following vaccination of vibrio cholerae adhesion molecule against shigella flexneri infection.

Introduction: Previous studies have shown 37.8 kDa pili subunit protein of Vibrio cholerae and 49.8 kDa pili subunit protein of Shigella flexneri can act as an adhesion molecule to initiate infection. These molecules also have the ability to agglutinate blood. The present study assessed mucosal and systemic immunity following vaccination using 37.8 kDa V. cholerae and protection against S. flexneri. Subjects and Methods: Haemagglutination test was performed after purification of V. cholerae protein, followed by an anti-haemagglutination test. The intestinal weight and colony count were used to validate the protective effect on balb/c mice which were divided into the naive group, Shigella-positive control group, Vibrio-positive control group, V. cholerae infected-Vibrio-vaccinated group and S. flexneri-infected-Vibrio-vaccinated group. Th17, Treg, interleukin (IL) IL-17A, ß-defensin and secretory-immunoglobulin A (s-IgA) were also measured to determine the systemic and mucosal immunity after vaccination. Results: The haemagglutination and anti-haemagglutination tests showed that the 37.8 kDa protein could inhibit 49.8 kDa of the S. flexneri pili subunit. Decreased intestinal weight and colony count of vaccinated group compared to naive group also support cross reaction findings. Vaccination also generates higher level of Th17, Treg, IL-17A, ß-defensin and s-IgA significantly. Conclusions: 37.8 kDa subunit pili can act as a homologous vaccine candidate to prevent V. cholerae and S. flexneri infection.

Effect of Nigella sativa Ethanol Extract on the Nitric Oxide Content and Renal Arteriole Diameter of a Pre-eclampsia Mouse Model.

OBJECTIVE: The aim of this study was to investigate the effect of Nigella sativa ethanol extract on nitric oxide (NO) levels and renal arteriole diameter of a pre-eclampsia mouse model. MATERIALS AND METHODS: This experimental study was conducted using a post-test only control group design. Thirty BALB/c mice were divided into six groups: one negative control (normal pregnant mice), one positive control (pre-eclampsia model), and four groups of pre-eclampsia mice treated with varying doses of N. sativa (500 mg/kg body weight (BW)/day, 1000 mg/kg BW/day, 1500 mg/kg BW/day, and 2000 mg/kg BW/day). Ethanol extract of N. sativa was given for 5 days. Data were analyzed using analysis of variance. RESULTS: We detected significant differences in NO levels between the pre-eclampsia mouse model and those given the ethanol extract. The latter had NO levels of 85.77±4.47 µM (500 mg/kg BW/day), 189.04±6.01 µM (1000 mg/kg BW/day), 226.56±2.13 µM (1500 mg/BW/day), and 207.98±4.74 µM (2000 mg/kg BW/day). The mean renal arteriole diameter showed significant differences among the treatment groups with N. sativa doses of 1000, 1500, and 2000 mg (15.15±2.21b µm, 16.35±2.52b µm, and 15.76±3.03b µm, respectively). CONCLUSION: N. sativa ethanol extract treatment increases NO levels and enlarges renal arteriole diameter of a pre-eclampsia mouse model in a dose-dependent manner.