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.

Human ACE-2, MCP1 and micro-RNA 146 as Novel Markers for COVID- 19 Affection and Severity.

BACKGROUND & AIMS: Coronavirus disease - 2019 (COVID-19) is a major pandemic that causes high morbidity and mortality rates. AIM OF THIS STUDY: to detect the relations between many risk factors, ACE-2, MCP-1, Micro RNA 146 gene expression, and COVID-19 infection and disease severity. METHODS: This study was carried out on 165 cases of COVID-19 and 138 controls. ACE2 and MCP1 levels were measured in COVID-19 cases and control by ELISA and micro-RNA-146 expression by PCR. RESULTS: We found an increased blood level of ACE2 and MCP1 in COVID- 19 patients than in healthy persons and a significant down-regulation of micro-RNA 146 gene expression in cases than in controls. There was a significant correlation between increased blood level of ACE2, regulation of micro-RNA 146 gene expression and severity of lung affection, a significant correlation was found between increased blood level of MCP1 and thrombosis in COVID-19 patients. Neurological complications were significantly correlated with more viral load, more ACE2 blood level, and down regulation of micro RNA146 expression. CONCLUSION: High viral load, increased blood level of ACE2, and down-regulation of micro-RNA 146 expression are associated with more severe lung injury and the presence of neurologic complications like convulsions and coma in COVID-19 Egyptian patients.

Clinical and mycological investigations of post-COVID-19 acute invasive fungal sinusitis.

Objectives: An increased incidence of acute invasive fungal sinusitis associated with the recent COVID-19 pandemic has been observed, which is considered a public health concern. This study aims to detect the incidence, risk factors, causative agents, clinical presentations, outcomes, and susceptibility rate of various antifungals. Methods: In this cross-sectional cohort study, a total of 30 patients showing acute invasive fungal rhinosinusitis following a COVID-19 infection were investigated. Histopathological biopsies, culture identification, and molecular confirmation of the causative agents were conducted. The demographic data, risk factors, clinical presentations, treatment regimen and its outcomes, and efficacy of antifungals were listed and analyzed. Results: A total of 30 cases with a mean age of 59.6 ± 11.9 years were included. Diabetes mellitus was the most recorded comorbidity with a rate of 86.7%, whereas most of the patients received corticosteroids. The mycological examination confirmed the existence of Mucor (Rhizopus oryzae) and Aspergillus (Aspergillus niger) in 96.7% and 3.3% of the cases, respectively. Various stages of sinonasal involvement (ethmoid, maxillary, sphenoid, and inferior turbinate) represented 100%, 83.3%, 66.7%, and 86.7% of the cases, respectively. Headache and facial pain, ophthalmoplegia, visual loss, and blindness represented 100%, 66.7%, 90%, and 53.3% of the cases, respectively. All the cases were simultaneously treated with surgical debridement and amphotericin B. Moreover, R. oryzae was susceptible to it, whereas A. niger was sensitive to voriconazole, resulting in a survival rate of 86.7% (26/30). The R. oryzae and A. niger isolates were proven to be sensitive to acetic acid, ethyl alcohol, formalin, and isopropyl alcohol. Conclusions: In patients with COVID-19, the diagnosis of acute invasive fungal sinusitis and prompt treatment with antifungal medicine and surgical debridement are important in achieving better outcomes and survival rates. Level of Evidence: 4.

Association of Forkhead Box O3a (FOXO3a) single - nucleotide polymorphism with bronchial asthma in Egyptian population.

Asthma is a common chronic inflammatory condition with a highly complex genetic predisposition and environmental factors which play an important role in its development. Polymorphism in FOXO3a transcription factor has been linked to a number of inflammatory and respiratory diseases such as bronchiolitis and idiopathic pulmonary fibrosis suggesting that it may be implicated in the pathogenesis of asthma. This study aimed to investigate FOXO3a SNP (rs13217795) association with bronchial asthma and its degree of severity in adult Egyptian population. This case control study included 60 asthmatic patients and 40 apparently healthy controls. Peripheral blood samples were collected from all participants. Genotyping was performed using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). The study revealed high frequency of the mutant TT genotype of FOXO3a gene in asthma patients (51.7%) than controls (12.5%) with OR= 7.48 & 95% CI (2.58-21.71) (P˂0.05) and in severe cases (41.9%) compared to mild and moderate cases (25.8% and12.5%, respectively). T allele frequency showed significant statistical association with asthma, OR= 12.40, 95% CI (5.65-27.19) (P˂0.05). However, there was no association between T allele and disease severity. The high frequency of the mutant TT genotype among patients and sever cases may indicates that FOXO3a rs13217795 C>T single nucleotide polymorphism can be considered as a risk factor in development and severity of asthma.