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The current outbreak of coronavirus-associated disease characterized as coronavirus disease 19 (COVID-19) marked its existence in late 2019. Since then, it has been a concern for human health and safety. Coronaviruses (CoVs) are known to cause a wide range of diseases; out of which common cold and pneumonia occur in human beings. In this review, we elaborate on the basic characteristics, structure, variants mutations and the pathological attributes of SARS-CoV 2. We also discuss the interaction of the viral proteins with the host cell receptor known as the ACE2 receptor and various therapeutics for the treatment of disease.

The novel coronavirus (SARS-CoV-2) outbreak has started taking away the millions of lives worldwide. Identification of known and approved drugs against novel coronavirus disease (COVID-19) seems to be an urgent need for the repurposing of the existing drugs. So, here we examined a safe strategy of using approved drugs of SuperDRUG2 database against modeled membrane protein (M-protein) of SARS-CoV-2 which is essential for virus assembly by using molecular docking-based virtual screening. A total of 3639 drugs from SuperDRUG2 database and additionally 14 potential drugs reported against COVID-19 proteins were selected. Molecular docking analyses revealed that nine drugs can bind the active site of M-protein with desirable molecular interactions. We therefore applied molecular dynamics simulations and binding free energy calculation using MM-PBSA to analyze the stability of the compounds. The complexes of M-protein with the selected drugs were simulated for 50 ns and ranked according to their binding free energies. The binding mode of the drugs with M-protein was analyzed and it was observed that Colchicine, Remdesivir, Bafilomycin A1 from COVID-19 suggested drugs and Temozolomide from SuperDRUG2 database displayed desirable molecular interactions and higher binding affinity towards M-protein. Interestingly, Colchicine was found as the top most binder among tested drugs against M-protein. We therefore additionally identified four Colchicine derivatives which can bind efficiently with M-protein and have better pharmacokinetic properties. We recommend that these drugs can be tested further through in vitro studies against SARS-CoV-2 M-protein.

Pulmonary fibrosis is characterized by damage to the epithelial cells and alveolar-capillary basement membrane. The increased expression levels of transforming growth factor β (TGF-β) and TGF-β-receptor-1 induced differentiation of lung fibroblasts to myofibroblasts, an alarming sign and considered the hallmark event development of pulmonary fibrosis. In the current study, the stability of phytochemicals of

This study exclusively focused on the potential application of an inexpensive and sustainable waste macro-algal biomass as an adsorbent for biosorption of copper ions from aqueous medium. After extraction of agar from brown macro-marine algaeGelidiella acerosa, the residual biomass without any further treatment was used as an adsorbent for the expulsion of copper from wastewater. Physicochemical parameters of biosorption like initial pH, initial concentration of Cu(II) solution and biosorbent dosage were optimized using response surface methodology. The maximum copper biosorption potential of 96.36% was observed at optimum conditions of pH of 5.31, initial concentration of 23.87 mg/l and biosorbent dosage of 0.41 g/l. Adopting FTIR and SEM techniques, the surface morphological features of biosorbent were studied. The pseudo-second-order kinetic model was found to be a proper approach to describe biosorption kinetics. All these results confirmed that spentG.acerosacould be considered as an efficient, eco-friendly and economic alternative for Cu(II) removal from aqueous solution.

Biosurfactants are amphiphilic compounds produced by bacteria either extracellularly or as a part of the cell membrane. Biosurfactants have had a profound impact on medical and pharmaceutical biotechnology. In our previous work, we isolated a new biosurfactant produced by Acinetobacter indicus M6 which reduces the surface tension of water from 72.0 to 39.8 mN/m and which showed thermophilic, halophytic and acidophilic stability. The chemical nature was found to be a class of glycolipoprotein. Here, our research presents the extracted biosurfactant’s anti-proliferative activity against lung cancer cells (A549), and anti-microbial and anti-biofilm activity against MRSA. The anti-tumour activity of biosurfactant against lung cancer cells was evaluated in terms of cell viability at different concentrations. The results showed a decrease in the percentage of lung cancer viable cells with increasing biosurfactant concentrations and incubation time, with a significant decrease being observed at 200 µg/ml concentration leading to cell proliferation inhibition at G1 phase. Treatment of biofilms for seven days at 500 µg/ml resulted in up to 82.5% biofilm disruption. Keywords: Biosurfactant, Acinetobacter indicus M6, Cell viability, Anti-proliferative activity, Anti-biofilm activity

The immunoglobulin (Ig) binding proteins of Staphylococcus aureus namely staphylococcal protein A (SpA) and staphylococcal binder of immunoglobulin (Sbi) are responsible for false positives during immunoassays. Avian IgY antibodies were reported to have no affinity to SpA and thus are safe for use in immunoassays. However, the behaviour of Sbi with IgY was not reported. The purpose of the present study is to evaluate the interactions between IgY antibodies and Sbi protein from different S. aureus strains. Initially, heterologous cloning and expression of complete sbi gene in Escherichia coli was undertaken. Recombinant Sbi protein was utilized to generate polyclonal anti-Sbi IgY and anti-Sbi antibodies in chicken and BALB/c mice respectively. Indirect ELISA and Western blotting were performed to evaluate the reactivity of anti-Sbi antibodies. Non-reducing PAGE followed by Western blotting and double-antibody sandwich dot-ELISA were performed to analyze the reactivity of IgY antibodies with recombinant Sbi and native Sbi from S. aureus strains. To avoid the possible interference of enzyme-conjugated secondary antibodies from mammalian sources, mouse anti-Sbi revealing antibodies were labeled with biotin so that streptavidin-HRP was used as developing reagent for chromogenic reaction. Sbi was highly immunogenic in chicken and mouse with antibody titers of 1:128,000 and 1:64,000 dilutions respectively. We observed that unimmunized IgY antibodies showed no affinity to either recombinant Sbi or native Sbi from S. aureus strains in Western blotting and double antibody sandwich ELISA. In view of these observations, we recommend that IgY antibodies are safe and free from false positives due to SpA and Sbi in immunoassays involving detection of S. aureus antigens/exotoxins.

A novel approach of cellulase-based co-culture producing bioethanol using low-cost nutrient medium has been employed for the study, and docking strategies provided the information of cellobiose and cellotetraose inhibitors during cellulase production. The Acinetobacter species was isolated from termite gut and confirmed in 16s rDNA analysis. SDS-PAGE reveals the molecular weight of purified cellulose was 45 kDa. Cellulose activity was achieved maximum of 1.2 IU/mL at 48th hour of incubation, and about 80% of polysaccharides were converted into simple sugars. The enzyme activity was optimum at different physiological conditions like temperature at 37 °C, pH 7.0 and with 4% concentration of banana peduncle extract powder. Upon reaching maximum cellulolytic activity of 0.594 IU/mL, the percentage of ethanol produced from cellulosic hydrolysate using S. cerevisiae reached maximum ethanol (18.3 g/L) during 48 h of incubation. Cellulase produced by Acinetobacter indicus KTCV2 strain exhibits a short incubation period (48 h) and produces cellulase in broader pH and temperature ranges. Banana peduncle offers a cheapest raw material composed of cellulosic biomass in agricultural practices, which served as a good substrate for the production of ethanol.

CRISPR genome editing utilizes Cas9 nuclease and single guide RNA (sgRNA), which directs the nuclease to a specific site in the genome and makes a double-stranded break (DSB). Design of sgRNA for CRISPR-Cas targeting, and to promote CRISPR adaptation, uses a regulatory mechanism that ensures maximum CRISPR-Cas9 system functions when a bacterial population is at highest risk of phage infection. Acinetobacter baumannii is the most regularly identified gram-negative bacterium infecting patients. Recent reports have demonstrated that the extent of diseases caused by A. baumannii is expanding and, in a few cases, now surpasses the quantity of infections caused by P. aeruginosa. Most Acinetobacter strains possess biofilm-forming ability, which plays a major role in virulence and drug resistance. Biofilm bacteria use quorum sensing, a cell-to-cell communication process, to activate gene expression. Many genes are involved in biofilm formation and the mechanism to disrupt the biofilm network is still not clearly understood. In this study, we performed in silico gene editing to exploit the AbaI gene, responsible for biofilm formation. The study explored different tools available for genome editing to create gene knockouts, selecting the A. baumannii AbaI gene as a target. Keywords: AbaI, Acinetobacter baumannii, CHOPCHOP, Crispr-cas9, sgRNA

Freshwater biota especially molluscs are more vulnerable because even the slightest change in the quality of their habitat possibly leads to their mortality. In addition, their importance is felt that particularly molluscs contribute toward carbon sequestering from the environment as atmospheric carbon gets deposited as carbonates in their shell which constitute 10% of their body mass. Hence, the molecular phylogenetic relationships of two families inhabiting freshwater streams viz., Ampullariidae and Viviparidae considering the genera dwelling in Indian subcontinent namely Pila globosa and Bellamya bengalensis using the partial nucleotide sequence of mitochondrial COI gene were of considerable importance in building conservational strategies. The phylogenetic analysis of ampullariid and viviparid species inferred that these two chosen families were paraphyletic and rooted on the genera of Cyclophoroidea and possibly with Saulea vitrea as the connecting link.

Background ATP-binding cassette (ABC) transporters constitute one of the largest transporter protein families and play a role in diverse biological processes. Results In the present study, bacteriocin isolated from the Enterococcus casseliflavus MI001 strain was identified as an ABC transporter ATP-binding protein. The optimal conditions for the production of bacteriocin were found to be at 35 °C, a pH 5.5, and an incubation time of 24 h. Purification was performed using ammonium sulphate precipitation, gel filtration, and DEAE ion exchange chromatography. The bacteriocin was purified with an eightfold purification scheme resulting with a specific activity of 15,000 AU/mg. The NMR spectrum of purified bacteriocin revealed the presence of amino acids, namely lysine, methionine, cysteine, proline, threonine, tryptophan, and histidine. Further, the bacteriocin ABC transporter showed antimicrobial activity against food spoilage microorganisms. Conclusions The ABC transporter ATP-binding protein could be used as a potential alternative for food preservation, and it may be considered as a bio-preservative agent in food processing industries.

Background: Gastrointestinal (GI) diseases are a major cause of emergency department visits requiring hospitalizations leading to considerable burden on global economy. Several factors contribute to the onset of gastrointestinal diseases such as pathogens (parasites, bacteria, virus, toxins etc.), autoimmune disorders and severe inflammation of intestine. Objective: One common feature among all these diseases is the dysentery and alteration of gut microbiota composition (gut dysbiosis). Apart from conventional therapies such as antibiotics and ORS supplementation, gut microbiota modulation with probiotic supplementation has emerged as a successful and healthy alternative in mitigating GI diseases. In this review our goal is to discuss the causes of gastrointestinal diseases and the present state of various therapeutic strategies such as probiotics as live biotherapeutics and Fecal Microbial Transplants (FMT’s). Conclusion: Several reports and clinical trials point out to the beneficial effects of probiotics in modulating the gut microbiota and improving the side effects of gastrointestinal diseases. Live biotherapeutics and FMT’s could be suitable and successful alternatives to conventional therapies in mitigating the gastrointestinal pathogens.

In the present work, a simple and straightforward method was developed to clone any PCR-amplified products into restriction sites that are very close, adjacent or overlapping in the expression vector. The novelty of the methodology involves a crucial primer-designing step by adding appropriate overhangs to the 5' ends of primers based on the multiple cloning sites (MCS) (polylinker) region of expression vector. After PCR amplification, actual cloning is performed not in adjacent RE sites, but in sites that are little distant in the MCS. However, the sites lost during this cloning step are maintained intact since they are provided by the cloned PCR product (through the primer overhangs). Gene for green fluorescent protein (GFP) was cloned and expressed employing this strategy to demonstrate its simplicity. This method is highly useful for vector modification without losing the restriction sites present in the MCS.

Staphylococcus aureus is an important pathogen responsible for a variety of diseases ranging from mild skin and soft tissue infections, food poisoning to highly serious diseases such as osteomyelitis, endocarditis, and toxic shock syndrome. Proper diagnosis of pathogen and virulence factors is important for providing timely intervention in the therapy. Owing to the invasive nature of infections and the limited treatment options due to rampant spread of antibiotic-resistant strains, the trend for development of vaccines and antibody therapy is increasing at rapid rate than development of new antibiotics. In this article, we have discussed elaborately about the host-pathogen interactions, clinical burden due to S aureus infections, status of diagnostic tools, and treatment options in terms of prophylaxis and therapy.

There is an increasing demand for the generation of recombinant pharmaceutical proteins for a wide array of therapeutic applications. In comparison to bacterial, yeast and animal cells, the production of recombinant proteins in plants with economic and therapeutic importance has only started recently. The most important prerequisite of any expression systems is that it should be simple and inexpensive. In this regard, plant-based expression has emerged an as accepted alternative to conventional expression platforms due to economic feasibility, rapid scalability, higher stability of recombinant proteins, safety due to lack of harmful substances (human, animal pathogens and pyrogens) and capability of producing proteins with desired secondary modifications. Heterologous expression using plants has played a pivotal role in the development of a myriad of recombinant proteins, including neutraceuticals and monoclonal antibodies being utilized in various therapeutic approaches. This paper presents an overview about the current status, various strategies and advantages of pharmaceutical protein production in plant expression systems. We also present a summary of expression of therapeutic monoclonal antibodies, vaccines, clinical trials and the regulatory aspects of plant-based expression. Furthermore, the challenges encountered in plant expression such as costs associated with existing purification strategies are discussed.