Aberrant DNA Methylation in Human Hepatocellular Carcinoma

Essential epigenetic mechanism i.e., DNA methylation in humans is being continuously acknowledged as a hallmark of various cancer. Hypo-methylation and CpG hyper methylation considered early events in cancer development hence; their understanding will provide us new tools for diagnosis. Hypo-methylation of repetitive sequences associated with genomic instability and may cause changes in the local chromatin environment and disrupt gene expression that contributes to carcinoma development. Additionally, hyper methylation of the promoter region of genes, including p15, p16, RASSF1A, accumulates during cancer development, which can influence the process of angiogenesis, DNA repair, regulation of cell cycle, apoptosis as well as tumour cell invasion. The methylation process mediated via DNA methyltransferases; hence, the variation in these enzymes may lead to hepatocellular carcinoma (HCC). Oncogenes activation and tumour suppressor genes inactivation during the growth and development of HCC, promotor hyper-methylation and hypo-methylation are the activaters. This review summarized recent DNA methylation information and role of aberrant methylation in cancer progression by using different research papers from NCBI between 2016-2020, that would be helpful in the diagnosis and treatment of hepatocellular carcinoma.  Keywords: DNA Methylation; CpG Island; Gene Expression; Hypo-Methylation; Hyper- Methylation; Review

In Vitro, D-Ribose and Formaldehyde Glycating Effects on Hen Egg White Lysozyme

Background: Glycation causes severe damage to the protein structure, instigating different diseases like cataracts, nephropathy, vasculopathy, retinopathy, atherosclerosis, neurodegenerative disease, diabetes, and age-dependent complications. Formaldehyde, a pollutant present in human habitation, is produced endogenously or exogenously during cooking or incinerating wood, paints, furniture, chipboards, fabric etc. Its higher concentrations can cause cell damage that promotes the formation of DNA/Protein cross-links. The present study aimed to evaluate the glycating effects of formaldehyde on hen egg white lysozyme in comparison with known glycating agent D-ribose. Methods: In this, in-vitro study, hen egg white lysozyme (HEWL) glycation with different concentrations of formaldehyde (0.25mM, 0.5mM, 1mM and 2mM) and D-ribose (0.01mM, 0.05mM, 0.1mM and 0.5mM) was examined using two different experimental conditions: concentration and time duration. Further cross-linking of protein was also analysed using SDS-PAGE technique. Results: Glycation of HEWL treated with formaldehyde increased with increasing concentrations (0.25mM, 0.5mM, 1mM and 2mM) and time duration (1, 3, 7 and 15 days). Cross linking of HEWL showed visible glycation at 2mM concentration. Cross-linked HEWL products gave dimer at 0.25mM and 0.5mM and trimers at 1mM and 2mMat 3, 7 and 15days. However, compared to formaldehyde, D-ribose glycation at different concentrations (0.01mM, 0.05mM, 0.1mM and 0.5mM) did not show the prominent cross linking of protein. Conclusion: Formaldehyde was found to be a more potent glycating agent compared to D-ribose. Compared to D-ribose, formaldehyde can produce protein misfolding and can be used in clinical research to establish the role of formaldehyde in patients with diseases. Keywords: Formaldehyde; D-Ribose; Lysozyme; SDS PAGE

Inflammatory markers and COVID-19 disease progression

Background: The COVID-19 pandemic has resulted in a global humanitarian crisis. Despite ongoing research, transmission risks and many disease characteristics remained unclear. Most patients have displayed elevated levels of certain inflammatory markers, which we sought to investigate further in relation to disease severity. The aim of this study was to examine the correlation between inflammatory markers and the severity of COVID-19 among patients. Methods: We conducted a cross-sectional study from April to September 2020, involving 143 COVID-19 PCR-positive patients from Ziauddin Hospital. Electronic patient records provided data on demographics, clinical status, and laboratory results. Results: The majority of PCR-positive patients were elderly males with comorbidities such as diabetes and hypertension. Almost all patients exhibited increased levels of various inflammatory markers, with procalcitonin (97.2%) being the most common. Statistically significant differences were observed in the levels of TLC (p = 0.005), CRP (p = 0.001), LDH (p = 0.001), Ferritin (p = 0.001), D-dimer (p = 0.001), and procalcitonin (p = 0.028), in relation to COVID-19 severity. Conclusions: The data suggest a significant association between levels of inflammatory markers and COVID-19 severity. All markers, except procalcitonin, demonstrated a significant correlation with disease severity. These results could enhance our understanding of COVID-19 pathogenesis and help predict and manage severe cases

Protein expression profiling of nuclear membrane protein reveals potential biomarker of human hepatocellular carcinoma

Abstract Background Complex molecular events lead to development and progression of liver cirrhosis to HCC. Differentially expressed nuclear membrane associated proteins are responsible for the functional and structural alteration during the progression from cirrhosis to carcinoma. Although alterations/ post translational modifications in protein expression have been extensively quantified, complementary analysis of nuclear membrane proteome changes have been limited. Deciphering the molecular mechanism that differentiate between normal and disease state may lead to identification of biomarkers for carcinoma. Results Many proteins displayed differential expression when nuclear membrane proteome of hepatocellular carcinoma (HCC), fibrotic liver, and HepG2 cell line were assessed using 2-DE and ESI-Q-TOF MS/MS. From the down regulated set in HCC, we have identified for the first time a 15 KDa cytochrome b5A (CYB5A), ATP synthase subunit delta (ATPD) and Hemoglobin subunit beta (HBB) with 11, 5 and 22 peptide matches respectively. Furthermore, nitrosylation studies with S-nitrosocysteine followed by immunoblotting with anti SNO-cysteine demonstrated a novel and biologically relevant post translational modification of thiols of CYB5A in HCC specimens only. Immunofluorescence images demonstrated increased protein S-nitrosylation signals in the tumor cells and fibrotic region of HCC tissues. The two other nuclear membrane proteins which were only found to be nitrosylated in case of HCC were up regulated ATP synthase subunit beta (ATPB) and down regulated HBB. The decrease in expression of CYB5A in HCC suggests their possible role in disease progression. Further insight of the functional association of the identified proteins was obtained through KEGG/ REACTOME pathway analysis databases. String 8.3 interaction network shows strong interactions with proteins at high confidence score, which is helpful in characterization of functional abnormalities that may be a causative factor of liver pathology. Conclusion These findings may have broader implications for understanding the mechanism of development of carcinoma. However, large scale studies will be required for further verification of their critical role in development and progression of HCC

Additional file 2 of Angiotensin-converting enzyme 2 (ACE2) polymorphisms and susceptibility of severe SARS-CoV-2 in a subset of Pakistani population

Supplementary Table 1: ACE2 haplotypes, haplotype frequency and permutation test for all haplotypes organized in block 1 and block

Additional file 1 of Angiotensin-converting enzyme 2 (ACE2) polymorphisms and susceptibility of severe SARS-CoV-2 in a subset of Pakistani population

Supplementary Figure: 1 (a): Gel doc for ACE2 demonstrating the bands for PCR products