Background and Objectives: Immunocompromised patients are considered a high-risk group to develop mycobacterial infections with either pulmonary and/or extra-pulmonary diseases. Low-cost and low-density (LCD) DNA-microarray is considered an easy and efficient method to detect typical and atypical Mycobacterial species. Materials and methods: Thirty immunocompromised patients were recruited to provide their clinical specimens (sputum, serum, urine and lymph node aspirate). Both Real-Time PCR and LCD-microarray techniques were performed and compared to the conventional methods of Ziehl-Neelsen (ZN) staining and Lowenstein Jensen (LJ) culturing. Results and conclusion: Mycobacterium tuberculosis complex (MTBC) was detected in all 30 clinical specimens (100% sensitivity) by Real-Time PCR and LCD-microarray. Additionally, co-infection with 4 atypical species belonging to mycobacteria other than tuberculosis (MOTT) was identified in 7 sputum specimens. These atypical mycobacterial species were presented as M. kansasii 10% (n=3), M. avium complex 6.6% (n=2), M. gordanae 3.3% (n=1) and M. peregnium 3.3% (n=1). This study documents the presence of certain species of atypical mycobacteria among immunocompromised patients in Egypt. To the best of our knowledge, this is the first detection of M. peregenium among clinical specimen in Egypt.

Although many vaccines have been approved for COVID-19, the continuing high incidence motivates the importance of drug repositioning. Among COVID-19 multiple complications, severe pneumonia and thromboembolism-related death are the most aggressive. We aimed to monitor the mechanisms by which SARS-CoV-2 led to severe illness and death to explore targets for therapies. Our objective was achieved by determining the cut-off of the novel biomarker human platelet membrane glycoproteins IIb/IIIa (CD41/CD61) (GPIIb/IIIa) in microparticle free plasma separated from peripheral blood samples of COVID-19 patients and healthy subjects using the ELISA technique. In comparison with the control, we observed an elevated level of GPIIb/IIIa in COVID-19 patients, especially those with severe illness including severe pneumonia and pulmonary embolism. Our data indicate the relevance of determination of GPIIb/IIIa as a diagnostic and prognostic biomarker for haematological complications including platelet aggregation and pulmonary embolism in COVID-19 patients suffering from severe illness. We conclude that determination of GPIIb/IIIa level by an easy, reliable, and low-cost ELISA kit helps in early diagnosis of haematological complications in COVID-19 patients and may help in improving the clinical outcomes and treatment of COVID-19 patients. Adding GPIIb/IIIa inhibitors (synthetic or natural products) to the treatment protocol of COVID-19 patients may add benefit in improving the clinical outcomes of COVID-19 patients.

Introduction. Tuberculosis (TB) is a great public health problem in developing countries such as Egypt. Genotyping of Mycobacterium tuberculosis isolates has a prominent role in the field of TB prevention. Aim. This study aimed to evaluate real-time PCR using Minor Groove Binder (MGB) probes and to identify circulating lineages/sub-lineages of M. tuberculosis and their transmission patterns. Hypothesis. We hypothesize that MIRU-VNTR technique is efficient in identifying circulating M. tuberculosis lineages in Egypt. Methodology. Fifty sputum specimens positive for acid-fast bacilli were included. Isoniazid (INH) resistance was detected using the 1 % proportion method. Real-time PCR using MGB-probes was used for simultaneous detection of TB infection and INH resistance. Partial sequencing of the katG gene was used to confirm INH resistance results. A standard 15 Mycobacterial Interspersed Repetitive Unit Variable Number Tandem Repeat (15-MIRU-VNTR) approach was used for genotyping through the MIRU-VNTRplus online platform. Results. Only seven specimens showed phenotypic resistance to INH. M. tuberculosis was detected in all samples, while a mutation in the katG gene codon 315 was detected only in five samples, which were also phenotypically INH-resistant. Sequencing of the katG gene showed codon 315 mutation genotypically and phenotypically in the five INH-resistant isolates. Molecular genotyping of M. tuberculosis isolates revealed that the majority of isolates (26/50, 52 %) belonged to the S family of lineage_4. A low clustering rate (2 %) was observed among our isolates. According to the Hunter-Gaston Discriminatory Index (HGDI), 11 MIRU-VNTR loci were highly or moderately discriminative, while four loci were less polymorphic. Conclusion. MIRU-VNTR genotyping revealed a low clustering rate with a low recent transmission rate of M. tuberculosis strains in Alexandria, Egypt.

Background. To date (14 January 2022), the incidence and related mortality rate of COVID-19 in America, Europe, and Asia despite administrated of billions doses of many approved vaccines are still higher than in Egypt. Epigenetic alterations mediate the effects of environmental factors on the regulation of genetic material causing many diseases. Objective. We aimed to explore the methylation status of HeyL promoter, a downstream transcription factor in Notch signal, an important regulator of cell proliferation and differentiation blood, pulmonary epithelial, and nerves cells. Methods. Our objective was achieved by DNA sequencing of the product from methyl-specific PCR of HeyL promoter after bisulfite modification of DNA extracted from the blood samples of 30 COVID-19 patients and 20 control health subjects and studying its association with clinical-pathological biomarkers. Results. We found that the HeyL promoter was partial-methylated in Egyptian COVID-19 patients and control healthy subjects compared to full methylated one that was published in GenBank. We identified unmethylated CpG (TG) flanking the response elements within HeyL promoter in Egyptian COVID-19 patients and control healthy subjects vs. methylated CpG (CG) in reference sequence (GenBank). Also, we observed that the frequency of partial-methylated HeyL promoter was higher in COVID-19 patients and associated with aging, fever, severe pneumonia, ageusia/anosmia, and dry cough compared to control healthy subjects. Conclusion. We concluded that hypomethylated HeyL promoter in Egyptian population may facilitate the binding of transcription factors to their binding sites, thus enhancing its regulatory action on the blood, pulmonary epithelium, and nerves cells in contrast to full methylated one that was published in GenBank; thus, addition of demethylating agents to the treatment protocol of COVID-19 may improve the clinical outcomes. Administration of therapy must be based on determination of methylation status of HeyL, a novel prognostic marker for severe illness in COVID-19 patients.