OBJECTIVETo study aberrant DNA methylation potentially resulting in changes in the expression of cancer-related genes as a possible epigenetic mechanism for cadmium carcinogenesis.

METHODSGenomic DNA isolated from CdCl(2)-transformed BALB/c-3T3 cells was digested with Mse1 (methylation non-sensitive) alone or with Mse1 and BstU1 (methylation sensitive). The resulting DNA was analyzed for aberrant methylation using PCR-based technique-Methylation-Sensitive Restriction Fingerprinting (MSRF). Several DNA fragments differentially methylated in the transformed cells identified by MSRF were confirmed by Southern hybridization analysis using the aberrantly methylated DNA fragments as the probes.

RESULTSAberrant DNA methylation was identified in the transformed cells. DNA sequencing and sequence similarity analysis identified one of the aberrantly methylated DNA fragments as the p16 tumor suppressor gene.

CONCLUSIONDNA hypermethylation is known to result in gene silencing, it appears that hypermethylation of p16 gene may represent a possible epigenetic mechanism for Cd-induced cell transformation and carcinogenesis.

Animals ; BALB 3T3 Cells ; Blotting, Southern ; Cadmium ; toxicity ; Cell Transformation, Neoplastic ; CpG Islands ; DNA Methylation ; Genes, p16 ; Mice ; Restriction Mapping

Malaria is a life-threatening disease caused by protozoan Plasmodium species. Plasmodium falciparum is the deadliest species. Reducing and eliminating malaria burden are linked to most of the Sustainable Development Goals (SDG), central to SDG3 targeting the end of malaria by 2030. This study was aimed at assessing the Management of malaria and prevalence of P. falciparum kelch-13 among febrile patients in selected Government Hospitals in Nigeria. Malaria patients (399) attending outpatient clinics of the Hospitals between August, 2019 and January, 2021, were enlisted in the study, following ethical approval and informed consents. Blood (5mL) was collected from patients for microscopic and molecular investigation of malaria parasite. DNA extraction, PCR amplification, BLAST, and alignment were performed. Plasmodium resistance to Artemether/lumefantrine was determined by PCR amplification of extracted DNA using Kelch-13 gene primer. Data obtained were subjected to One-way Analysis of Variance and Linear Regression. The VapA gene primer amplified 55 (68.75%) out of the 80 DNA extracts tested. Twenty-five strains of P. falciparum belonging to 3 clades phylogenetically were identified and they showed evolutionary relationships with others. Plasmodium falciparum resistant Kelch-13 gene was detected in 70% of the isolates. This study observed a high prevalence of resistant gene to ACT drugs in the study area. Monitoring the effectiveness of ACTs must be done routinely to ensure timely changes in National treatment policies.

Objective: To evaluate and compare the effects of bevacizumab, mitoinycin-C (MMC), 5-fluorouracil (5-FU), and triamcinolone acetonide (TA) on the viability of cultured human Tenon's capsule fibroblasts (cHTF) in vitro.

Methods: Human Tenon's fibroblasts (HTF) were harvested and cultured in a Roswell-Park-Memorial 1-Institute (RPMI) media. MMC, 5-FU, bevaciz. umab, and TA were administered to the cHTF at 3-fold decreasing concentrations starting from 20 ug, 5 mg, 25 mg, and 4 mg respectively. A negative control/untreated group containing RPMI media only was included in the study. Fibroblast cell viability was assessed using resazurin fluorim etric assay. Half¬maximal inhibitory concentration (IC50) was computed for agents which showed significant decrease in cHTF viability compared to the untreated group.

Results: There was no significant difference in cH IF viability between the untreated control group compared to 5-FU (p=0.97), bevacizumab (p=0.10), and TA (p=0.06) groups. Mitomycin-C showed a significant decrease in cHTF viability (p<0.001) which was dose dependent. The IC50 of MMC was computed at 12.16 ug using the prism software.

Conclusion: Mitomycin-C demonstrated dose-dependent decrease in viability of cultured human Tenon's fibroblasts. 5-FU, bevacizumab, and triamcinolone did not show this effect.

Key Words: Mitomycin-C, 5-fluorouracil, Bevaciz. umab, Tria. mcinolone acetonide, Fibroblast, Trabeculectomy

Human ; Male ; Female ; .humans ; Mitomycin ; Triamcinolone Acetonide ; Trabeculectomy ; Resazurin ; Bevacizumab ; Fluorouracil ; Cell Survival ; Control Groups ; Inhibitory Concentration 50 ; Tenon Capsule ; Xanthenes ; Oxazines ; Antibodies, Monoclonal, Humanized ; Fibroblasts ; Software