Introduction: The current monkeypox (MPX) outbreak, caused by the monkeypox virus (MPXV), has turned into a global concern, with over 59,000 infection cases and 23 deaths worldwide.Objectives: Herein, we aimed to exploit robust immunoinformatics approach, targeting membrane-bound, enveloped, and extracellular proteins of MPXV to formulate a chimeric antigen. Such a strategy could similarly be applied for identifying immunodominant epitopes and designing multi-epitope vaccine ensembles in other pathogens responsible for chronic pathologies that are difficult to intervene against.Methods: A reverse vaccinology pipeline was used to select 11 potential vaccine candidates, which were screened and mapped to predict immunodominant B-cell and T-cell epitopes. The finalized epitopes were merged with the aid of suitable linkers, an adjuvant (Resuscitation-promoting factor), a PADRE sequence (13 aa), and an HIV TAT sequence (11 aa) to formulate a multivalent epitope vaccine. Bioinformatics tools were employed to carry out codon adaptation and computational cloning. The tertiary structure of the chimeric vaccine construct was modeled via I-TASSER, and its interaction with Toll-like receptor 4 (TLR4) was evaluated using molecular docking and molecular dynamics simulation. C-ImmSim server was implemented to examine the immune response against the designed multi-epitope antigen.Results and discussion: The designed chimeric vaccine construct included 21 immunodominant epitopes (six B-cell, eight cytotoxic T lymphocyte, and seven helper T-lymphocyte) and is predicted non-allergen, antigenic, soluble, with suitable physicochemical features, that can promote cross-protection among the MPXV strains. The selected epitopes indicated a wide global population coverage (93.62%). Most finalized epitopes have 70%–100% sequence similarity with the experimentally validated immune epitopes of the vaccinia virus, which can be helpful in the speedy progression of vaccine design. Lastly, molecular docking and molecular dynamics simulation computed stable and energetically favourable interaction between the putative antigen and TLR4.Conclusion: Our results show that the multi-epitope vaccine might elicit cellular and humoral immune responses and could be a potential vaccine candidate against the MPXV infection. Further experimental testing of the proposed vaccine is warranted to validate its safety and efficacy profile.

The present experiment was designed to isolate bacterial strains from the brick kiln soil and to check the activity and enzyme kinetics of amylase from these isolates. The bacterial colonies were isolated from soil samples through the serial dilution method. The bacterial isolates were identified through morphological, electron microscopic and molecular analysis. The 16S ribosomal RNA sequences of the isolates IR-1, IR-2, IR-3, IR-8, and IR-9 showed high similarities with Bacillus tequilensis, Bacillus paramycoides, Proteus alimentorum, Bacillus wiedmannii, and Pseudomonas aeruginosa, respectively. All of the bacterial isolates showed a positive catalase activity except IR-9. Furthermore, the isolates showed variable antagonistic effects against different bacterial pathogens. All of the strains produced indole acetic acid (IAA), and the concentrations increased in the presence of tryptophan application. The isolates showed the amylase enzyme activity and maximum activity of isolates was achieved in 4% starch concentration. The IR-9 isolate showed the highest amylase activity of 5.9 U/ml. The Vmax values of the extracellular amylase from different bacterial isolates ranged between 12.90 and 50.00 IU ml−1. The lowest Km value of 6.33 mg starch was recorded for IR-8 and the maximum Kcat value of 2.50 min−1 was observed for IR-3. The amylase activity of the isolates was significantly affected by a range of different incubation time, temperature, and pH values. Further tests are required before the potential utilization of these isolates for amylase production, and in the biopesticide and biofertilizer applications.

The current coronavirus disease-2019 (COVID-19) pandemic has triggered a worldwide health and economic crisis. The severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) causes the disease and completes its life cycle using the RNA-dependent RNA-polymerase (RdRp) enzyme, a prominent target for antivirals. In this study, we have computationally screened ∼690 million compounds from the ZINC20 database and 11,698 small molecule inhibitors from DrugBank to find existing and novel non-nucleoside inhibitors for SARS-CoV-2 RdRp. Herein, a combination of the structure-based pharmacophore modeling and hybrid virtual screening methods, including per-residue energy decomposition-based pharmacophore screening, molecular docking, pharmacokinetics, and toxicity evaluation were employed to retrieve novel as well as existing RdRp non-nucleoside inhibitors from large chemical databases. Besides, molecular dynamics simulation and Molecular Mechanics/Generalized Born Surface Area (MM/GBSA) method were used to investigate the binding stability and calculate the binding free energy of RdRp-inhibitor complexes. Based on docking scores and significant binding interactions with crucial residues (Lys553, Arg557, Lys623, Cys815, and Ser816) in the RNA binding site of RdRp, three existing drugs, ZINC285540154, ZINC98208626, ZINC28467879, and five compounds from ZINC20 (ZINC739681614, ZINC1166211307, ZINC611516532, ZINC1602963057, and ZINC1398350200) were selected, and the conformational stability of RdRp due to their binding was confirmed through molecular dynamics simulation. The free energy calculations revealed these compounds possess strong binding affinities for RdRp. In addition, these novel inhibitors exhibited drug-like features, good absorption, distribution, metabolism, and excretion profile and were found to be non-toxic. The compounds identified in the study by multifold computational strategy can be validated in vitro as potential non-nucleoside inhibitors of SARS-CoV-2 RdRp and holds promise for the discovery of novel drugs against COVID-19 in future.

Zinc (Zn) is an important micronutrient with vital biological roles and its deficiency causes serious complications both in plants and human cells. The present research aims to introduce a sustainable intervention to improve zinc bioavailability, partitioning and biofortification in wheat genotypes through the use of zinc solubilizing bacteria. Three bacterial strains; Pseudomonas lurida (DJ4), Pseudomonas aeruginosa (DJ5) and Pseudomonas fluorescens (DJ13), previously isolated from a forest soil in Dir, Pakistan with zinc solubilization potential were used during this study. The maximum of 242.34 and 108.08 µg·l⁻¹ gluconic and glucuronic acid, respectively, was produce by the bacterial strain DJ13 in the presence of zinc oxide and consequently, a decrease in medium pH from 7.0 to 4.63 was noted. Further, root colonization potential of these bacterial strains with wheat genotypes (Atta Habib, Sirin and Pak China) was confirmed by scanning electron microscope (SEM). Similarly, the zinc uptake and translocation potential of the eight wheat genotypes under no zinc, ZnSO₄.7H₂O supplementation or inoculation with DJ13 was evaluated in a pot experiment in greenhouse. Wheat genotypes Zincol 2016, Sirin and Atta Habib attained higher grain zinc content after DJ13 inoculation (57.87, 63.55 and 60.47 mg·kg⁻¹ respectively). Genetic differences in zinc uptake, translocation and grain accumulation in wheat genotypes under different growth conditions were noted to be because of differential expression of ZIP transporters. High expression levels of TaZIP1, TaZIP4, TaZIP13, and TabZIP1 were correlated with increased root uptake, TaZIP1 and TabZIP1 were correlated with zinc translocation and TaZIP5 with grain accumulation. In conclusion, this research demonstrated the capacity to solubilize zinc and establish a synergistic interaction with wheat by specific bacterial strains, particularly Pseudomonas fluorescens (DJ13), significantly enhancing zinc uptake, translocation, and mobilization. These strains also modulated the expression of ZIP genes, leading to increased zinc partitioning in wheat grains. Consequently, these rhizobacteria present a highly effective biofertilizer strategy for zinc biofortification of grains and the optimization of wheat growth and development.

| An experiment was conducted to study the effect of foliar application of CaCl 2 on alleviation of the adverse effects of water stress on maize (CV. Azam). The experiment was conducted in completely randomized design (CRD) using pots in a glass house and comprised of 2 irrigations (irrigated and water stress) and three concentrations of CaCl 2 (5, 10 and 15 mM CaCl 2 .2H 2 O) as foliar spray on maize seedlings 20 days after emergence (DAE). Water was sprayed as untreated control on maize seedlings 20 DAE. After foliar application, the plants were regularly irrigated upto one week and water stress was imposed by withholding water from half of the pots for 20 days. Data were recorded on the relative water content (RWC), proline, sugar content and transcripts abundance of Lhcb2 gene in the selected leaves. Application of CaCl 2 increased the RWC, proline and sugar content as well as the Lhcb2 expression in the leaves under both irrigation conditions; however, the effect was more prominent under water stress. In the control plants, water stress decreased RWC (-57%) and Lhcb2 transcript abundance (-44.37%) but increased proline (9.02 fold) and sugar content (3.42 fold) after 20 days. Significantly higher RWC (82.00 and 55.67%), proline (4.13 and 13.29 nmol·g -1 FW) and sugar content (51.19 and 102.00 mg·g -1 FW) under irrigated and water stressed conditions, respectively, was noted when 10 mM CaCl 2 was applied as foliar spray. Furthermore, 10 mM CaCl 2 application enhanced the Lhcb2 transcripts by 18 and 62% under irrigated and water stress conditions, respectively. These results demonstrate the importance of CaCl 2 foliar application on the improvement in the water relations and biochemical adaptation, resulting in improvement in the performance of plants under water deficit conditions. These results also demonstrated that senescence, which is a type of cell death program that could be inappropriately activated during drought, was also delayed in the supplemented seedlings as highlighted by a high RWC and expression of the Lhcb2 gene.