Elucidating the EXOSC3-IRE1α interaction: a convergent study incorporating computational, in vitro and in vivo studies.

The Unfolded protein response (UPR) is an adaptive signalling pathway which is triggered by accumulation of unfolded/misfolded protein in ER lumen. The UPR consist of three transmembrane proteins-IRE1α, PERK and ATF6 that sense ER stress which leads to activation and downstream signaling from ER lumen to cytosol to restore homeostasis. IRE1α is an evolutionary conserved arm of UPR and acts as an interaction platform for many potential proteins that become activated under ER stress conditions. We investigated potential partners of IRE1 α through MS studies and found EXOSC3 as one of the binding partner of IRE1α. Exosomal complex proteins have 3'5' exonuclease properties (EXOSC3) that play an important role in mRNA surveillance. This property of exosomal proteins coincides with IRE1α ribonuclease activities and its mechanism of action is similar to that of IRE1α-RIDD pathway which degrades any unstable mRNA that disrupts cellular homeostasis. At the same time, studies have shown that knockdown of EXOSC3 causes ER stress in human cells, so we speculated that there might be a functional crosstalk between IRE1α and EXOSC3 under ER stress conditions. Therefore, we employed computational tools to predict and explore the stability and dynamics of the IRE1α-EXOSC3 complex. The analysis indicates that IRE1α and EXOSC3 exhibit potential interaction with the involvement of ScanNet, predicting binding pockets between the two proteins. Further, the interaction was validated via co-immunoprecipitation and yeast two-hybrid assays, thus suggesting EXOSC3 as a component of the UPRosome complex. Hence, this functional crosstalk might influence the dynamic functional output of IRE1α.Communicated by Ramaswamy H. Sarma.

Comparative assessment of SARS CoV2-associated mortalities in 3 COVID waves and related risk factors: The South Kashmir's experience.

Introduction: Severe acute respiratory syndrome coronavirus 2 causing coronavirus disease 2019 (COVID-19) pandemic have spurted in three major waves in India at different times and had different levels of severity in different waves. The objective of our study was to determine the comparative mortality rate in three COVID-19 waves and determine the factors associated with mortality. Methods: We identified a cohort of 1,132 COVID-19 patients who were admitted between April 14, 2020 and February 08, 2022 at our center. All the admitted patients with positive COVID-polymerase chain reaction were included in the study. Sample characteristics were determined by screening age, sex, socio-economic status, occupation, symptomatology of COVID, patient status on admission, baseline investigations, comorbidities, medical history, oxygen dosage needed during admission, the span of hospital stay, diagnosis, and vitals such as blood pressure, pulse, and oxygen saturation. All the data were procured from an institutional database. Results: In total, 1,132 patients included in the study, the mean age was 65.08 ± 12.29 and 56% were males. The affliction rate was 42.13% in >60 years, 29.5% in 46-60 years, 20.8% in 31-45 years, and 7.4% in 30 years' group. In the first, second, and third waves of COVID-19, the mortality rates were 13.21%, 23.53%, and 11.39%, respectively. Among the comorbidities, mortality rates were proportionately higher in those with hypertension (6.7%), and diabetes (5.5%), than those with chronic obstructive pulmonary disease (3.3%), chronic kidney disease (CKD) (1.5%), heart disease (1.6%), and malignancy (0.2%). Conclusion: We identify the peaked mortalities in the second encounter which was predicted by age, comorbidities such as hypertension, and diabetes.

An Investigation of the Antiviral Potential of Phytocompounds against Avian Infectious Bronchitis Virus through Template-Based Molecular Docking and Molecular Dynamics Simulation Analysis.

Vaccination is widely used to control Infectious Bronchitis in poultry; however, the limited cross-protection and safety issues associated with these vaccines can lead to vaccination failures. Keeping these limitations in mind, the current study explored the antiviral potential of phytocompounds against the Infectious Bronchitis virus using in silico approaches. A total of 1300 phytocompounds derived from fourteen botanicals were screened for their potential ability to inhibit the main protease, papain-like protease or RNA-dependent RNA-polymerase of the virus. The study identified Methyl Rosmarinate, Cianidanol, Royleanone, and 6,7-Dehydroroyleanone as dual-target inhibitors against any two of the key proteins. At the same time, 7-alpha-Acetoxyroyleanone from Rosmarinus officinalis was found to be a multi-target protein inhibitor against all three proteins. The potential multi-target inhibitor was subjected to molecular dynamics simulations to assess the stability of the protein-ligand complexes along with the corresponding reference ligands. The findings specified stable interactions of 7-alpha-Acetoxyroyleanone with the protein targets. The results based on the in silico study indicate that the phytocompounds can potentially inhibit the essential proteins of the Infectious Bronchitis virus; however, in vitro and in vivo studies are required for validation. Nevertheless, this study is a significant step in exploring the use of botanicals in feed to control Infectious Bronchitis infections in poultry.

Ribavirin inhibits the replication of infectious bursal disease virus predominantly through depletion of cellular guanosine pool.

Introduction: The antiviral activity of different mutagens against single-stranded RNA viruses is well documented; however, their activity on the replication of double-stranded RNA viruses remains unexplored. This study aims to investigate the effect of different antivirals on the replication of a chicken embryo fibroblast-adapted Infectious Bursal Disease virus, FVSKG2. This study further explores the antiviral mechanism utilized by the most effective anti-IBDV agent. Methods: The cytotoxicity and anti-FVSKG2 activity of different antiviral agents (ribavirin, 5-fluorouracil, 5-azacytidine, and amiloride) were evaluated. The virus was serially passaged in chicken embryo fibroblasts 11 times at sub-cytotoxic concentrations of ribavirin, 5-fluorouracil or amiloride. Further, the possible mutagenic and non-mutagenic mechanisms utilized by the most effective anti-FVSKG2 agent were explored. Results and Discussion: Ribavirin was the least cytotoxic on chicken embryo fibroblasts, followed by 5-fluorouracil, amiloride and 5-azacytidine. Ribavirin inhibited the replication of FVSKG2 in chicken embryo fibroblasts significantly at concentrations as low as 0.05 mM. The extinction of FVSKG2 was achieved during serial passage of the virus in chicken embryo fibroblasts at ≥0.05 mM ribavirin; however, the emergence of a mutagen-resistant virus was not observed until the eleventh passage. Further, no mutation was observed in 1,898 nucleotides of the FVSKG2 following its five passages in chicken embryo fibroblasts in the presence of 0.025 mM ribavirin. Ribavarin inhibited the FVSKG2 replication in chicken embryo fibroblasts primarily through IMPDH-mediated depletion of the Guanosine Triphosphate pool of cells. However, other mechanisms like ribavirin-mediated cytokine induction or possible inhibition of viral RNA-dependent RNA polymerase through its interaction with the enzyme's active sites enhance the anti-IBDV effect. Ribavirin inhibits ds- RNA viruses, likely through IMPDH inhibition and not mutagenesis. The inhibitory effect may, however, be augmented by other non-mutagenic mechanisms, like induction of antiviral cytokines in chicken embryo fibroblasts or interaction of ribavirin with the active sites of RNA-dependent RNA polymerase of the virus.

A multiepitope vaccine candidate against infectious bursal disease virus using immunoinformatics-based reverse vaccinology approach.

Infectious bursal disease virus is the causative agent of infectious bursal disease (Gumboro disease), a highly contagious immunosuppressive disease of chicken with a substantial economic impact on small- and large-scale poultry industries worldwide. Currently, live attenuated vaccines are widely used to control the disease in chickens despite their issues with safety (immunosuppression and bursal atrophy) and efficiency (breaking through the maternally-derived antibody titer). To overcome the drawbacks, the current study has, for the first time, attempted to construct a computational model of a multiepitope based vaccine candidate against infectious bursal disease virus, which has the potential to overcome the safety and protection issues found in the existing live-attenuated vaccines. The current study used a reverse vaccinology based immunoinformatics approach to construct the vaccine candidate using major and minor capsid proteins of the virus, VP2 and VP3, respectively. The vaccine construct was composed of four CD8+ epitopes, seven CD4+ T-cell epitopes, 11 B-cell epitopes and a Cholera Toxin B adjuvant, connected using appropriate flexible peptide linkers. The vaccine construct was evaluated as antigenic with VaxiJen Score of 0.6781, immunogenic with IEDB score of 2.89887 and non-allergenic. The 55.64 kDa construct was further evaluated for its physicochemical characteristics, which revealed that it was stable with an instability index of 16.24, basic with theoretical pI of 9.24, thermostable with aliphatic index of 86.72 and hydrophilic with GRAVY score of -0.256. The docking and molecular dynamics simulation studies of the vaccine construct with Toll-like receptor-3 revealed fair structural interaction (binding affinity of -295.94 kcal/mol) and complex stability. Further, the predicted induction of antibodies and cytokines by the vaccine construct indicated the possible elicitation of the host's immune response against the virus. The work is a significant attempt to develop next-generation vaccines against the infectious bursal disease virus though further experimental studies are required to assess the efficacy and protectivity of the proposed vaccine candidate in vivo.

Isolated Avulsion Fractures of Lesser Trochanter in Adolescents - A Case Series and Brief Literature Review.

INTRODUCTION: Adolescent apophyseal avulsion injuries related to sports is common around the pelvis and proximal femur. Amid them, avulsions of the lesser trochanter are uncommon as compared to other sites. The growth plates in children are weaker than the unossified bone and tendons to which they attach. As a result, with sudden violent muscular contraction, avulsion fracture of the apophysis occurs. The periosteum remains attached to the avulsed fragment preventing gross displacement. Treatment is usually symptomatic with good outcomes. CASE SERIES: We present a series of three cases of isolated avulsions of lesser trochanter. Patients presented with pain in groin and limp following sprinting and fall. Radiographs revealed the diagnosis, and we proceeded with conservative management in all patients since the displacement was not significant. Patients recovered fully with no long-term complaints. Indications for surgery are not well defined, and we believe conservative treatment has good functional outcomes. CONCLUSION: In patients with painful limp after sporting activities, there should be a high index of suspicion for avulsion fractures. Radiographs should be done, and other pathologies were excluded. Symptomatic treatment is recommended with a gradual return to sports after at least 3 months.

First description of the male caste of the Himalayan endemic ant Lasiusalienoflavus Bingham, 1903 (Hymenoptera: Formicidae), with re-description of the female and queen castes.

The present paper provides a description of the male caste and re-description of the worker and queen castes of the poorly known ant species Lasiusalienoflavus Bingham, 1903. This species has hitherto been reported only from the Himalayas, and the present data are also based on specimens collected in the north-western part of the mountain range. Likewise other Himalayan ants, this species also shows restricted distribution, which suggests a rather high degree of endemism (45%) of this group in the Himalayas.