Epigenetic alterations of miR-155 and global DNA methylation as potential mediators of ochratoxin A cytotoxicity and carcinogenicity in human lung fibroblasts.

Ochratoxin A (OTA) is a well-known mycotoxin that adversely affects different human cells. Inhalational exposure to OTA and subsequent pulmonary diseases have been previously reported, yet its potential carcinogenicity and underlying molecular mechanisms have not been fully elucidated. This study aimed to evaluate the OTA-induced cytotoxicity and the epigenetic changes underlying its potential carcinogenicity in fetal lung fibroblast (WI-38) cells. OTA cytotoxicity was assessed by MTT assay; RT-qPCR was used to determine the expression of BAX, BCL-2, TP53, and miR-155, while ELISA was used for measuring 5-methyl cytosine percentage to assess global DNA methylation in OTA-treated versus control cells. WI-38 cells demonstrated sensitivity to OTA with IC50 at 22.38 µM. Though BAX and Bcl-2 were downregulated, with low BAX/BCL-2 ratio, and TP53 was upregulated, their fold changes showed decline trend with increasing OTA concentration. A significant dose-dependent miR-155 upregulation was observed, with dynamic time-related decline. Using subtoxic OTA concentrations, a significant global DNA hypermethylation with significant dose-dependent and dynamic alterations was identified. Global DNA hypermethylation and miR-155 upregulation are epigenetic mechanisms that mediate OTA toxicity on WI-38 cells. BAX downregulation, reduced BAX/BCL-2 ratio together with miR-155 upregulation indicated either the inhibition of TP53-dependent apoptosis or a tissue specific response to OTA exposure. The aforementioned OTA-induced variations present a new molecular evidence of OTA cytotoxicity and possible carcinogenicity in lung fibroblast cells.

Preliminary Study of sCD14 and sCD163 as Predictors of Disease Severity and ICU Admission in COVID-19: Relation to Hematological Parameters, Blood Morphological Changes and Inflammatory Biomarkers.

Background and Objectives: Research supports the role of monocyte/macrophage activation in COVID-19 immunopathology. This study aimed to evaluate sCD14 and sCD163 - the monocyte activation markers - and to investigate their relation to hematological parameters and blood morphology in COVID-19 infection. Methods: This is a case-control study that included 70 COVID-19 patients. Patients were subdivided into two groups: 23 severely diseased ICU-admitted patients and another group of 47 non-ICU-admitted patients. sCD163 and sCD14 levels were determined using ELISA. Results: sCD163 and sCD14 showed significantly higher levels in sera of patients compared to the control group, with significantly higher levels of sCD163 in ICU-admitted patients than non-ICU admitted patients. Receiver operating characteristic curve analysis demonstrated the usefulness of sCD163 with a cut-off value of 734 ng/mL as a potential marker to discriminate between ICU and non-ICU admitted COVID-19 patients (sensitivity of 81.16%; specificity of 96.67% and positive predictive value of 98% with area under the curve of 0.930). sCD163 levels showed a positive correlation with total white blood cells, absolute neutrophilic count, Neutrophil/Lymphocyte ratio, and a negative correlation with platelet count. sCD14 levels positively correlated with D-dimer values associated with a shift to the left and neutrophilic toxic granulations in blood morphology. Conclusion: High sCD163 and sCD14 levels, hematological parameters, and blood morphology reflect monocyte activation in COVID-19 infection. sCD163 is a potential marker of disease severity. These findings support further study of therapeutics targeting macrophage activity in COVID-19 patients with high sCD163 levels.