The B-13 cell is a readily expandable rat pancreatic acinar-like cell that differentiates on simple plastic culture substrata into replicatively-senescent hepatocyte-like (B-13/H) cells in response to glucocorticoid exposure.

Background/Aim: DNA methylation is the most studied epigenetic modification in cancer. Ten-eleven translocation enzymes (TET) catalyze the oxidation of 5methylcytosine (5-mC) to 5-hydroxymethylcytosine (5-hmC) in the DNA. In the current research, we aimed to evaluate the role of 5-hmC and TET enzymes in non-small cell lung cancer (NSCLC) patients and their possible association with outcomes. Patients and Methods: ELISA was used to measure the 5-hmC levels in genomic DNA and qRT-PCR was used to evaluate TET1, TET2, and TET3 mRNAs expression levels in NSCLC tissues and their paired normal controls. Results: The levels of 5-hmC were significantly lower in NSCLC tissues than in normal tissues, with a mean ±SD of 0.28±0.37 vs. 1. 84±0.58, respectively (t=22.77, p<0.0001), and this reduction was correlated with adverse clinical features. In addition, all TET genes were significantly down-regulated in NSCLC tissues in comparison to their matched normal tissues. The mean±SD level of p<0.0001), TET2-mRNA level in NSCLC was 5.25±2.78 vs. 9.52±1.01 in normal tissues (t=14.48, p<0.0001), p<0.0001). Downregulation of TET genes was correlated with poor clinical features. Conclusion: 5-HmC levels as well as TET1, TET2, and TET3 mRNA levels were reduced in NSCLC tissues. The reduced levels of 5-hmC and TET mRNAs were associated with adverse clinical features, suggesting that the level of 5-hmC may serve as a valuable prognostic biomarker for NSCLC.

The rat pancreatic AR42J-B13 (B-13) cell line differentiates into non-replicative hepatocyte-like (B-13/H) cells in response to glucocorticoid. Since this response is dependent on an induction of serine/threonine protein kinase 1 (SGK1), this may suggest that a general pivotal role for SGK1 in hepatocyte maturation. To test this hypothesis, the effects of expressing adenoviral-encoded flag tagged human SGK1F (AdV-SGK1F) was examined at 3 stages of human induced pluripotent stem cell (iPSC) differentiation to hepatocytes. B-13 cells infected with AdV-SGK1F in the absence of glucocorticoid resulted in expression of flag tagged SGK1F protein; increases in β-catenin phosphorylation; decreases in Tcf/Lef transcriptional activity; expression of hepatocyte marker genes and conversion of B-13 cells to a cell phenotype near-similar to B-13/H cells. Given this demonstration of functionality, iPSCs directed to differentiate towards hepatocyte-like cells using a standard protocol of chemical inhibitors and mixtures of growth factors were additionally infected with AdV-SGK1F, either at an early time point during differentiation to endoderm; during endoderm differentiation to anterior definitive endoderm and hepatoblasts and once converted to hepatocyte-like cells. SGK1F expression had no effect on differentiation to endoderm, likely due to low levels of expression. However, expression of SGK1F in both iPSCs-derived endoderm and hepatocyte-like cells both resulted in promotion of cells to an hepatoblast phenotype. These data demonstrate that SGK1 expression promotes an hepatoblast phenotype rather than maturation of human iPSC towards a mature hepatocyte phenotype and suggest a transient role for Sgk1 in promoting an hepatoblast state in B-13 trans-differentiation to B-13/H cells.

Background: Hyperglycemic patients are at a high risk of COVID-19 severity. Neutrophils have been considered critical effector cells in COVID-19 development. Vitamin D deficiency is prevalent in hyperglycemic patients and was found to adversely associate with the neutrophil count. Aim: The goal of this work was to evaluate the characteristics of diabetic and pre-diabetic COVID-19 patients and discovered changes in neutrophils and their correlation, if any, with disease clinical presentation. Patients and Methods: The study included total of (514) Covid-19 positive patients confirmed by PCR and recruited from the Prince Mohammad Bin Abdulaziz Hospital in Riyadh, Saudi Arabia. Patient’s clinical characteristics were collected for all patients. Laboratory tests include HbA1c, neutrophil count, C-reactive protein (CRP), erythrocyte sedimentation rate (ESR), ferritin, D- dimer, 25 hydroxy vitamin D (25(OH)D), and folate. Results: The results found that 286 patients (55.6%) were diabetic, 77 patients (15%) were pre-diabetic and 151 (29.4%) were normoglycaemic. A significant difference was exhibited regarding the neutrophil count and inflammatory factors of COVID-19 severity. Furthermore, the neutrophil count was found to be directly correlated with the severity monitoring biochemical markers for Covid-19: CRP, ESR, ferritin, and D-dimer and inversely associated with vitamin D levels in diabetic and pre-diabetic patients. Conclusion: Our findings highlight the change of neutrophils in COVID-19 diabetic and pre-diabetic patients that was found to correlate positively with CRP, ESR, ferritin, and D-dimer, and negatively with 25(OH)D, but their correlation with the clinical presentation of the disease need further large investigations.