Mutational analysis of hemoglobin genes and functional characterization of detected variants, through in-silico analysis, in Pakistani beta-thalassemia major patients.

Thalassemia is one of the most prevalent genetic disorders worldwide. The present study aimed to explore the mutational spectrum of all hemoglobin (HB) encoding genes and to identify the potentially damaging and pathogenic variants in the beta (ß)-thalassemia major patients and thalassemia minor carriers of Southern Punjab, Pakistan. A total of 49 ß-thalassemia major patients and 49 carrier samples were screened for the identification of HBA1, HBA2, HBB, HBD, HBE1, HBG1 and HBG2 variants by NGS. PCR was performed for the amplification of HB encoding genes and the amplified product of 13 patients and 7 carrier samples were processed for the Sanger sequencing. Various bioinformatics tools and databases were employed to reveal the functional impact and pathogenicity potential of the observed variants. Results depicted a total of 20 variants of HB-related genes by NGS and 5 by Sanger sequencing in thalassemia patients. While 20 variants by NGS and 3 by Sanger were detected in carriers. Few known genetic variants of HB-encoding genes are being reported for the first time in Pakistani thalassemia patients and carriers. However, two novel HBB variants c.375A>C (p.P125P) and c.*61T>G and a novel variant of HBE1 (c.37A>T (p.T13S)) were also documented. Pathogenicity analysis predicted the pathogenic potential of HBB variants (c.47G>A (p.W16*), c.27-28insG (p. S10fs), and c.92+5G>C) for ß thalassemia. The study of functional impact indicated that these HBB variants result in the premature termination of translation leading to the loss of functional ß-globin protein. It is therefore suggested that the pathogenic HBB variants, identified during present study, can be employed for the diagnosis, carrier screening, and planning therapy of thalassemia.

Cytokinin Production by Azospirillum brasilense Contributes to Increase in Growth, Yield, Antioxidant, and Physiological Systems of Wheat (Triticum aestivum L.).

Plant growth-promoting rhizobacteria are known to associate with several cereal crops. The rhizobacterium exerts its function by synthesizing diverse arrays of phytohormones, such as cytokinin (Ck). However, it is difficult to determine the plant growth promotion when a bacterium produces many different kinds of phytohormones. Therefore, to assess the involvement of Ck in growth promotion and activation of antioxidant and physiological systems, we set up this experiment. Wheat seeds (Triticum aestivum L.) were inoculated with Azospirillum brasilense RA-17 (which produces zeatin type Ck) and RA-18 (which failed to produce Ck). Results showed that seed inoculation with RA-17 significantly improved growth and yield-related parameters compared with RA-18. The activity of enzymes, proline contents, and endogenous hormonal levels in wheat kernels were improved considerably with RA-17 than with RA-18. Strain RA-17 enhanced grain assimilation more than strain RA-18 resulting in a higher crop yield. These results suggest that microbial Ck production may be necessary for stimulating plant growth promotion and activating antioxidant and physiological systems in wheat.

Public knowledge, attitudes, and practices towards COVID-19 in Pakistan: A cross-sectional study.

In response to the recent COVID-19 pandemic, each country has adopted different measures for its control, for instance, complete lockdown, smart lockdown based on hotspot analysis, and/or strict movement control on their residents. The effectiveness of such controls depends largely on public knowledge, attitude, and practices towards this novel virus. The prime objective of this study is to examine the knowledge, attitudes, and practices of university students and graduates in Pakistan regarding COVID-19. For this purpose, cross-sectional data is collected from 1789 participants through an online survey conducted between July 12 and August 10. The survey instrument consists of demographic characteristics, 13 items on knowledge, 6 items on attitudes, and 3 items on practices, modified from a previously published questionnaire on COVID-19. Our statistical analyses are based on descriptive statistics, chi-square tests, and analysis of variance (ANOVA). The overall correct response rate of the knowledge section is 73%, which is comparatively lower than similar studies in the region. The majority of participants are optimistic about the successful control of COVID-19 in Pakistan, and following the instructions issued by the government of Pakistan. However, a bulk of participants showed skepticism and no trust in the health facilities of hospitals in Pakistan, which could be the potential explanation of falling confirmed cases in Pakistan. In addition, a low knowledge score is found to be significantly associated with low confidence in successful control of COVID-19, and socially undesirable practices. The results suggest an urgent need for health education programs to hold optimistic attitudes and rebuilding of trust of the general public in health facilities in local hospitals to avoid the possible next wave of COVID-19 in Pakistan.

Positive Selection Drives the Adaptive Evolution of Mitochondrial Antiviral Signaling (MAVS) Proteins-Mediating Innate Immunity in Mammals.

The regulated production of filamentous protein complexes is essential in many biological processes and provides a new paradigm in signal transmission. The mitochondrial antiviral signaling protein (MAVS) is a critical signaling hub in innate immunity that is activated when a receptor induces a shift in the globular caspase activation and recruitment domain of MAVS into helical superstructures (filaments). It is of interest whether adaptive evolution affects the proteins involved in innate immunity. Here, we explore and confer the role of selection and diversification on mitochondrial antiviral signaling protein in mammalian species. We obtined the MAVS proteins of mammalian species and examined their differences in evolutionary patterns. We discovered evidence for these proteins being subjected to substantial positive selection. We demonstrate that immune system proteins, particularly those encoding recognition proteins, develop under positive selection using codon-based probability methods. Positively chosen regions within recognition proteins cluster in domains involved in microorganism recognition, implying that molecular interactions between hosts and pathogens may promote adaptive evolution in the mammalian immune systems. These significant variations in MAVS development in mammalian species highlights the involvement of MAVS in innate immunity. Our findings highlight the significance of accounting for how non-synonymous alterations affect structure and function when employing sequence-level studies to determine and quantify positive selection.