COVID-19 caused by SARS-CoV-2, is an international concern. This infection requires urgent efforts to develop new antiviral compounds. To date, no specific drug in controlling this disease has been...

Background: Functional constipation is a common and annoying gastrointestinal disorder among children worldwide in which the intestinal microbiota composition plays a fundamental role. This study aimed to compare the quantity of main intestinal Lactobacillus species in constipated children and healthy controls. Materials and Methods: Fecal samples were collected from 40 children fulfilling Rome IV criteria for functional constipation and 40 healthy volunteers. Quantitative real-time polymerase chain reaction (qPCR) method with species-specific primers was used to investigate seven main Lactobacillus species in fecal samples. Results: Lactobacillus strains of the patient group were different from the healthy controls, main differences being noticeable decrease in the population quantity of Lactobacillus reuteri (mean 10^2.61 CFU/gram feces) and lower abundance of Lactobacillus fermentum (p< 0.0001). Conclusion: Lactobacillus species might be quantitatively altered in childhood functional constipation. Our findings on the different species of Lactobacillus population showed significantly decreased quantity in the patient group compared with the healthy subjects.

Introduction. Herpes simplex virus type 1 (HSV-1) causes serious human disease and establishes a long-term latent infection. The latent form of this virus has shown to be resistant to antiviral drugs. Clustered Regularly Interspace Short Palindromic Repeats (CRISPR), an important tool in genome engineering composed of guide RNA (gRNA) and Cas9 nuclease that makes an RNA-protein complex to digest exclusive target sequences implementation of gRNA. Moreover, CRISPR-Cas9 system effectively suppresses HSV-1 infection by knockout of some viral genes. Material and methods. To survey the efficacy of Cas9 system on HSV-1 genome destruction, we designed several guide RNAs (gRNAs) that all packaged in one vector. Also, we performed a one-step restriction using BamHI and ESP3I enzymes. Results. CRISPR/Cas9 system targeted against the gD gene of HSV-1 was transfected into HEK-AD cells that moreover, revealed a significant reduction of HSV-1 infection by plaque assay and real-time PCR. Conclusion. The pCas-Guide-EF1a-GFP CRISPR Vector can create a fast and efficient method for gRNA cloning by restriction enzymes (ESP3I (BsmBI) and BamHI). Therefore, the CRISPR/Cas9 system may be utilized for the screening of genes significant for the HSV1 infection and the development of new strategies for targeted therapy of viral infections caused by HSV-1. Introduction. Herpes simplex virus type 1 (HSV-1) causes serious human disease and establishes a long-term latent infection. The latent form of this virus has shown to be resistant to antiviral drugs. Clustered Regularly Interspace Short Palindromic Repeats (CRISPR), an important tool in genome engineering composed of guide RNA (gRNA) and Cas9 nuclease that makes an RNA-protein complex to digest exclusive target sequences implementation of gRNA. Moreover, CRISPR-Cas9 system effectively suppresses HSV-1 infection by knockout of some viral genes. Material and methods. To survey the efficacy of Cas9 system on HSV-1 genome destruction, we designed several guide RNAs (gRNAs) that all packaged in one vector. Also, we performed a one-step restriction using BamHI and ESP3I enzymes. Results. CRISPR/Cas9 system targeted against the gD gene of HSV-1 was transfected into HEK-AD cells that moreover, revealed a significant reduction of HSV-1 infection by plaque assay and real-time PCR. Conclusion. The pCas-Guide-EF1a-GFP CRISPR Vector can create a fast and efficient method for gRNA cloning by restriction enzymes (ESP3I (BsmBI) and BamHI). Therefore, the CRISPR/Cas9 system may be utilized for the screening of genes significant for the HSV1 infection and the development of new strategies for targeted therapy of viral infections caused by HSV-1.