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Genetic engineering of allogeneic cell therapeutics that fully prevents rejection by a recipient's immune system would abolish the requirement for immunosuppressive drugs or encapsulation and support large-scale manufacturing of off-the-shelf cell products. Previously, we generated mouse and human hypoimmune pluripotent (HIP) stem cells by depleting HLA class I and II molecules and overexpressing CD47 (B2M-/-CIITA-/-CD47+). To determine whether this strategy is successful in non-human primates, we engineered rhesus macaque HIP cells and transplanted them intramuscularly into four allogeneic rhesus macaques. The HIP cells survived unrestricted for 16 weeks in fully immunocompetent allogeneic recipients and differentiated into several lineages, whereas allogeneic wild-type cells were vigorously rejected. We also differentiated human HIP cells into endocrinologically active pancreatic islet cells and showed that they survived in immunocompetent, allogeneic diabetic humanized mice for 4 weeks and ameliorated diabetes. HIP-edited primary rhesus macaque islets survived for 40 weeks in an allogeneic rhesus macaque recipient without immunosuppression, whereas unedited islets were quickly rejected.

Abstract Background Aspergillus flavus Link:Fr, an opportunistic fungus that produces aflatoxin, is pathogenic to maize and other oilseed crops. Aflatoxin is a potent carcinogen, and its presence markedly reduces the value of grain. Understanding and enhancing host resistance to A. flavus infection and/or subsequent aflatoxin accumulation is generally considered an efficient means of reducing grain losses to aflatoxin. Different proteomic, genomic and genetic studies of maize (Zea mays L.) have generated large data sets with the goal of identifying genes responsible for conferring resistance to A. flavus, or aflatoxin. Results In order to maximize the usage of different data sets in new studies, including association mapping, we have constructed a relational database with web interface integrating the results of gene expression, proteomic (both gel-based and shotgun), Quantitative Trait Loci (QTL) genetic mapping studies, and sequence data from the literature to facilitate selection of candidate genes for continued investigation. The Corn Fungal Resistance Associated Sequences Database (CFRAS-DB) (http://agbase.msstate.edu/) was created with the main goal of identifying genes important to aflatoxin resistance. CFRAS-DB is implemented using MySQL as the relational database management system running on a Linux server, using an Apache web server, and Perl CGI scripts as the web interface. The database and the associated web-based interface allow researchers to examine many lines of evidence (e.g. microarray, proteomics, QTL studies, SNP data) to assess the potential role of a gene or group of genes in the response of different maize lines to A. flavus infection and subsequent production of aflatoxin by the fungus. Conclusions CFRAS-DB provides the first opportunity to integrate data pertaining to the problem of A. flavus and aflatoxin resistance in maize in one resource and to support queries across different datasets. The web-based interface gives researchers different query options for mining the database across different types of experiments. The database is publically available at http://agbase.msstate.edu.

Introduction: Non Alcoholic Fatty Liver Disease (NAFLD) is one of the major leading causes of liver diseases, comprising a spectrum of conditions ranging from simple steatosis to cirrhosis. In the era of preventive medicine, it is of utmost importance to recognise the subset of NAFLD patients at high risk of progressing to liver cirrhosis. A newly emerging concept of myosteatosis is now suspected to be an early manifestation of NAFLD disease progression. Aim: To find the correlation between myosteatosis and liver fibrosis among patients with NAFLD. Materials and Methods: This was a cross-sectional study conducted in the Department of Radiodiagnosis and Department of Gastroenterology at Amala Institute of Medical Sciences in Thrissur, Kerala, India, from January 2021 to June 2022. A total of 57 subjects with Magnetic Resonance Imaging-proven (MRI) NAFLD were included in the study. Body weight and height were measured. Liver fat and myosteatosis were measured using the MRI-derived Proton Density Fat Fraction (PDFF) method {Iterative Decomposition of Water and Fat with Echo (IDEAL-IQ sequence)}. Liver fibrosis was assessed using 2D shear wave elastography. The proportion of myosteatosis and liver fibrosis among NAFLD patients was estimated. Partial correlation, controlling for gender, was evaluated using partial Spearman’s rho correlation coefficients. An Reciever Operating Characteristic (ROC) curve was plotted to assess muscle fat fraction in predicting liver fibrosis outcome among patients. Results: Out of the 57 subjects studied, 17 were females and 40 were males. The median Interquartile Range (IQR) age of the subjects was 43.0 (16.5). The median MRI hepatic fat fraction was 10.8. The median muscle PDFF in males was 8.4, and in females, it was 16.9. The median H-PDFF was 18.8. Myosteatosis correlated positively with liver fibrosis (r=0.558; p

ObjectiveLimb-girdle muscular dystrophies (LGMDs), one of the most heterogeneous neuromuscular disorders (NMDs), involves predominantly proximal-muscle weakness with >30 genes associated with different subtypes. The clinical-genetic overlap among subtypes and with other NMDs complicate disease-subtype identification lengthening diagnostic process, increases overall costs hindering treatment/clinical-trial recruitment. Currently seven LGMD clinical trials are active but still no gene-therapy-related treatment is available. Till-date no nation-wide large-scale LGMD sequencing program was performed. Our objectives were to understand LGMD genetic basis, different subtypes' relative prevalence across US and investigate underlying disease mechanisms.MethodsA total of 4656 patients with clinically suspected-LGMD across US were recruited to conduct next-generation sequencing (NGS)-based gene-panel testing during June-2015 to June-2017 in CLIA-CAP-certified Emory-Genetics-Laboratory. Thirty-five LGMD-subtypes-associated or LGMD-like other NMD-associated genes were investigated. Main outcomes were diagnostic yield, gene-variant spectrum, and LGMD subtypes' prevalence in a large US LGMD-suspected population.ResultsMolecular diagnosis was established in 27% (1259 cases; 95% CI, 26-29%) of the patients with major contributing genes to LGMD phenotypes being: CAPN3(17%), DYSF(16%), FKRP(9%) and ANO5(7%). We observed an increased prevalence of genetically confirmed late-onset Pompe disease, DNAJB6-associated LGMD subtype1E and CAPN3-associated autosomal-dominant LGMDs. Interestingly, we identified a high prevalence of patients with pathogenic variants in more than one LGMD gene suggesting possible synergistic heterozygosity/digenic/multigenic contribution to disease presentation/progression that needs consideration as a part of diagnostic modality.InterpretationOverall, this study has improved our understanding of the relative prevalence of different LGMD subtypes, their respective genetic etiology, and the changing paradigm of their inheritance modes and novel mechanisms that will allow for improved timely treatment, management, and enrolment of molecularly diagnosed individuals in clinical trials.

Phosphoglycerate kinase-1 (PGK1) deficiency is a rare X-linked disorder caused by pathogenic variants in the PGK1 gene. Complete loss-of-function variants have not been reported in this gene, indicating that residual enzyme function is critical for viability in males. Therefore, copy number variants (CNVs) that include single exon or multiple exon deletions or duplications are generally not expected in individuals with PGK1 deficiency. Here we describe a 64-year-old male presenting with a family history (three additional affected males) and a personal history of childhood-onset metabolic myopathy that involves episodes of muscle pain, stiffness after activity, exercise intolerance, and myoglobinuria after exertion. Biochemical analysis on a muscle biopsy indicated significantly reduced activity (15% compared to normal) for phosphoglycerate kinase (PGK1), a glycolytic enzyme encoded by PGK1. A diagnosis of PGK1 deficiency was established by molecular analysis which detected an approximately 886 kb deletion involving the polyadenylation site in the 3'UTR of the PGK1 gene. RNA analysis showed significantly reduced PGK1 transcript levels (30% compared to normal). This is the first deletion reported in the PGK1 gene and is the first pathogenic variant involving the 3'UTR polyadenylation site of this gene. Our report emphasizes the role of 3'UTR variants in human disorders and underscores the need for exploring noncoding regions of disease-associated genes when seeking a molecular diagnosis.

Abstract Background Patient satisfaction to Antenatal care services has traditionally been linked to the quality of services given and the extent to which specific needs are met. Even though data in this area was limited in Ethiopia, improving quality of care was one of the strategies in health sector development program IV. This study, therefore, attempted to assess client satisfaction to skilled antenatal care services in the study area. Methods and materials A cross-sectional facility based survey was conducted among women who were attending antenatal care clinic, using quantitative method triangulated with qualitative data collection. Participants were selected using systematic sampling method according to the flow pregnant women to the antenatal care clinics. The study was carried out in all functional public health centers in the district. During the survey, 405 women were interviewed. A logistic regression model was applied to control for confounders. Results Out of the total respondents, overall satisfied to skilled antenatal care services were about 277(68%). The most common specific component of antenatal care that had good-satisfaction by the respondents was “Privacy” at examination (81.7%). Most satisfied health education session was “Diet and nutrition” session (82.2%). Absence of sonar test, no doctor and long waiting time were commonest causes of dissatisfaction. Respondents who have > 2 previous antenatal care visit were 3 times more likely (AOR = 2.93; 95% CI, 1.21, 7.12) to have satisfaction to antenatal care services as compared to those with

Fungal endophytes are the critical organisms in the environment residing in the plant tissues without showing any harmful effects on its host life cycle. The finding of fungal endophytes in natural habitat has been insufficient due to some non-sporulating and non-culturable fungal endophytes by traditional method. Several investigations on fungal endophytes in plants have resulted in an excessive knowledge of the group. This review emphasis on the biology of fungal endophytes, their discovery, isolation, identification by morphological and molecular methods, production, purification and structure elucidation of the bioactive compounds.

Laccase is an effective biocatalyst for the degradation of persistent phenolic pollutants. However, its high cost and instability under harsh operational conditions hinder its applications. Hence a peptide/nucleotide-matrix-free laccase mimetic nanozyme would be attractive. Herein, MoS2/Cu is synthesized as a laccase mimic for catalytic oxidation of phenolic pollutants. Characterization of the nanozyme by SEM-EDX, XRD, and XPS confirmed the presence of multi-oxidation state copper on MoS2 which is consistent with the composition of natural laccase. The laccase mimetic activity of MoS2/Cu was examined by using 2,4-dichloro phenol as a substrate which oxidized to form a colored product with 4-aminoantipyrine (λmax = 510 nm). Surprisingly, MoS2/Cu showed 55.6% higher activity than pristine Cu NPs at pH 7, indicating the role of MoS2 as an effective scaffold. The catalytic reaction exhibited a lower Km (0.11 mM) than that reported for natural laccase. MoS2/Cu showed improved stability retaining 84.63% of its activity at 90 °C and lost only 10.93% of its activity after one month. Furthermore, it exhibited remarkable activity in the presence of different salts and alcohol stress. Notably, the nanozyme converted over 92% of phenolic substrates in 9 h. The results imply a robust performance for the remediation of phenolic compounds. [ABSTRACT FROM AUTHOR]

Introduction: Non Alcoholic Fatty Liver Disease (NAFLD) is one of the major leading causes of liver diseases, comprising a spectrum of conditions ranging from simple steatosis to cirrhosis. In the era of preventive medicine, it is of utmost importance to recognise the subset of NAFLD patients at high risk of progressing to liver cirrhosis. A newly emerging concept of myosteatosis is now suspected to be an early manifestation of NAFLD disease progression. Aim: To find the correlation between myosteatosis and liver fibrosis among patients with NAFLD. Materials and Methods: This was a cross-sectional study conducted in the Department of Radiodiagnosis and Department of Gastroenterology at Amala Institute of Medical Sciences in Thrissur, Kerala, India, from January 2021 to June 2022. A total of 57 subjects with Magnetic Resonance Imaging-proven (MRI) NAFLD were included in the study. Body weight and height were measured. Liver fat and myosteatosis were measured using the MRI-derived Proton Density Fat Fraction (PDFF) method {Iterative Decomposition of Water and Fat with Echo (IDEAL-IQ sequence)}. Liver fibrosis was assessed using 2D shear wave elastography. The proportion of myosteatosis and liver fibrosis among NAFLD patients was estimated. Partial correlation, controlling for gender, was evaluated using partial Spearman's rho correlation coefficients. An Reciever Operating Characteristic (ROC) curve was plotted to assess muscle fat fraction in predicting liver fibrosis outcome among patients. Results: Out of the 57 subjects studied, 17 were females and 40 were males. The median Interquartile Range (IQR) age of the subjects was 43.0 (16.5). The median MRI hepatic fat fraction was 10.8. The median muscle PDFF in males was 8.4, and in females, it was 16.9. The median H-PDFF was 18.8. Myosteatosis correlated positively with liver fibrosis (r=0.558; p<0.001). It also negatively correlated with hepatic steatosis (r=-0.321; p=0.02). A statistically significant correlation was not found between liver fat and liver fibrosis. An ROC curve was plotted to predict the liver fibrosis outcome by muscle fat fraction {Area Under Curve (AUC: 0.605; p-value: 0.204)}, which showed a sensitivity of 0.615 and a specificity of 0.389 at a cutoff score of 10.43. Conclusion: Myosteatosis positively correlated with liver fibrosis and negatively with liver steatosis. [ABSTRACT FROM AUTHOR]