Isothermal PCR for Feasible Molecular Diagnosis of Primary Toxoplasmosis in Women Recently Experienced Spontaneous Abortion

AIM: The current study aimed to assess the practicability of a simple loop-mediated isothermal amplification (LAMP) about real-time quantitative PCR to diagnose primary toxoplasmosis among high-risk pregnant women.METHODS: Cloned Toxoplasma samples were used to calculate the analytical sensitivity while specificity was assessed using pooled DNA samples extracted from other parasitic stages.RESULTS: Both techniques showed 100% sensitivity and specificity and then applied to detect recent Toxoplasma infection in peripheral blood of 77 IgG negative women out of a total 139 women lately experienced spontaneous abortion. The 2 techniques obtained positive results in 8 samples confirming primary toxoplasmosis.CONCLUSION: Generally, LAMP assay is a simple, cost-effective molecular technique can be completed in less than half an hour to diagnose primary Toxoplasma infection. The technique can be applied in a minimally equipped laboratory by ordinary workers to screen the vulnerable groups. Further analysis using larger samples with the quantitative approach is recommended to confirm the sensitivity of this emergent molecular technique

Molecular Diagnosis of Toxoplasmosis in Non Immune Pregnant Females

BACKGROUND AND AIM: Infection with the protozoan parasite Toxoplasma gondii has a worldwide distribution. Congenital infection is the most important part of the disease burden due to Toxoplasma infection in humans. Early diagnosis of maternal infection helps to prevent severe complications of toxoplasmosis. In the present study, three PCR assays (conventional, nested & quantitative) were evaluated for diagnosis of recent toxoplasmosis based on detection of Toxoplasma B1 gene.MATERIAL AND METHODS: The present study was carried out on 150 pregnant females who were serologically negative for anti-Toxoplasma IgG and IgM antibodies.RESULTS: The results revealed that out of 12 true positive cases (by 2 out of the 3 PCR protocols), 8 cases were positive by cPCR, 11 cases were positive by nPCR and 12 cases were positive by qPCR. Accurate estimation of genomic Toxoplasma DNA in positive samples was achieved by qPCR. In general, PCR assays offer a sensitive alternative of serological methods for diagnosis of recent maternal toxoplasmosis. In addition, qPCR decreases the risk of contamination of PCR products being a closed tube method and helps in estimation of infection load.CONCLUSIONS: We recommend screening of high-risk pregnant women by qPCR for early diagnosis of toxoplasmosis and proper management

SARS-CoV-2 susceptibility and COVID-19 disease severity are associated with genetic variants affecting gene expression in a variety of tissues

Variability in SARS-CoV-2 susceptibility and COVID-19 disease severity between individuals is partly due to genetic factors. Here, we identify 4 genomic loci with suggestive associations for SARS-CoV-2 susceptibility and 19 for COVID-19 disease severity. Four of these 23 loci likely have an ethnicity-specific component. Genome-wide association study (GWAS) signals in 11 loci colocalize with expression quantitative trait loci (eQTLs) associated with the expression of 20 genes in 62 tissues/cell types (range: 1:43 tissues/gene), including lung, brain, heart, muscle, and skin as well as the digestive system and immune system. We perform genetic fine mapping to compute 99% credible SNP sets, which identify 10 GWAS loci that have eight or fewer SNPs in the credible set, including three loci with one single likely causal SNP. Our study suggests that the diverse symptoms and disease severity of COVID-19 observed between individuals is associated with variants across the genome, affecting gene expression levels in a wide variety of tissue types

A first update on mapping the human genetic architecture of COVID-19

peer reviewe

Relation of cytokeratin 18-Apoptosense M30 to activity and fibrosis in chronic HCV patients

AbstractHepatitis C virus (HCV) is a progressive disease that may result in Chronic active hepatitis, cirrhosis, and hepatocellular carcinoma. Cytokeratin(CK)18 is an intermediary filament protein, expressed in hepatocytes, which is proteolytically cleaved during liver damage. M30 epitope of cytokeratin18(CK18-M30) is involved at different levels in apoptotic pathways. The aim of this study to investigate the diagnostic accuracy of cytokeratin 18-apoptosin M30 fragments (CK18-M30) as non-invasive method of liver fibrosis assessment and their correlation to METAVIR score. Serum concentrations of CK18-M30 were measured by ELISA in One hundred and sixty eight chronic hepatitis C patients (112 males and 56 females). Results showed that serum concentrations of CK18 were significantly increased in a stepwise fashion from A0 to A3 and from F0 to F4.There was highly significant positive correlation between CK18 and fibrosis stages (r = 0.881 & p < 0.001) and activity grades (r = 0.881 & p < 0.001). Also, serum CK18 correlated positively with serum levels of transaminases (r = 0.355 & p < 0.05). CK18 was correlated positively with viral level (r = 0.3 & p < 0.05). It was concluded that Elevated serum CK18-M30, as an indicator of enhanced apoptosis of hepatocytes, was increased and correlated significantly with inflammation severity, stages of fibrosis, transaminases (ALT) levels and viral load in chronic HCV patients. These findings may place CK18-M30 as a non-invasive marker of liver fibrosis and disease activity

Clinical spectrum of non alcoholic fatty liver disease in patients with chronic obstructive pulmonary disease

OBJECTIVE: The purpose of this study was to determine the prevalence of nonalcoholic fatty liver disease in a group of chronic obstructive pulmonary disease patients. MATERIAL AND METHODS: This study comprised 48 stable chronic obstructive pulmonary disease patients who were diagnosed and categorized using the Global Initiative for Chronic Obstructive Lung Disease 2017 criteria. The prevalence of nonalcoholic fatty liver disease in chronic obstructive pulmonary disease patients was determined using noninvasive biomarkers and imaging methods. Steatosis was detected using magnetic resonance mDIXON-Quant sequence imaging, while fibrosis was detected using the acoustic radiation force impulse and FIB-4 index. RESULTS: A total of 58.3% of the patients investigated had a fat level of 5%, and nearly a quarter of them had a fat content of 10% or more, and 45.8% of the patients studied had severe hepatic fibrosis. The Fibrosis-4 (FIB-4) index revealed advanced fibrosis in 18.75% of them. No statistically significant association was found between chronic obstructive pulmonary disease groups of studied patients and the presence of steatosis and fibrosis (≥F2) using acoustic radiation force impulse. The presence of fibrosis, however, was statistically significant linked with chronic obstructive pulmonary disease groups of examined patients using the FIB-4 index. γ-Glutamyl transferase and alkaline phosphatase levels were greater in Global Initiative for Chronic Obstructive Lung Disease 3/4 and C/D groups. CONCLUSION: Nonalcoholic fatty liver disease is a common comorbidity in chronic obstructive pulmonary disease and should be included in the list of chronic obstructive pulmonary disease comorbidities

Anti-Tumor Activity of Orally Administered Gefitinib-Loaded Nanosized Cubosomes against Colon Cancer

Gefitinib (GFT) is a tyrosine kinase inhibitor drug used as a first-line treatment for patients with advanced or metastatic non-small cell lung, colon, and breast cancer. GFT exhibits low solubility and hence low oral bioavailability, which restricts its clinical application. One of the most important trends in overcoming such problems is the use of a vesicular system. Cubosomes are considered one of the most important vesicular systems used to improve solubility and oral bioavailability. In this study, GFT cubosomal nanoparticles (GFT-CNPs) were prepared by the emulsification method. The selected formulation variables were analyzed and optimized by full factorial design and response surface methodology. Drug entrapment efficiency (EE%), transmission electron microscopy, particle size, polydispersity index, in vitro release and its kinetics, and the effect of storage studies were estimated. The chosen GFT-CNPs were subjected to further investigations as gene expression levels of tissue inhibitors of metalloproteinases-1 (TIMP-1) and matrix metalloproteinases-7 (MMP-7), colon biomarkers, and histopathological examination of colon tissues. The prepared GFT-CNPs were semi-cubic in shape, with high EE%, smaller vesicle size, and higher zeta potential values. The in vivo data showed a significant decrease in the serum level of embryonic antigen (CEA), carbohydrate antigen 19-9 (CA 19-9), and gene expression level of TIMP-1 and MMP-7. Histopathological examination showed enhancement in cancer tissue and highly decreased focal infiltration in the lamina propria after treatment with GFT-CNPs

COVID-19 Host Genetics Initiative. A first update on mapping the human genetic architecture of COVID-19

The COVID-19 pandemic continues to pose a major public health threat, especially in countries with low vaccination rates. To better understand the biological underpinnings of SARS-CoV-2 infection and COVID-19 severity, we formed the COVID-19 Host Genetics Initiative1. Here we present a genome-wide association study meta-analysis of up to 125,584 cases and over 2.5 million control individuals across 60 studies from 25 countries, adding 11 genome-wide significant loci compared with those previously identified2. Genes at new loci, including SFTPD, MUC5B and ACE2, reveal compelling insights regarding disease susceptibility and severity.</p