Search for RNA aptamers against non-structural protein of SARS-CoV-2: Design using molecular dynamics approach

Background Recent outbreak of deadly Severe Acute Respiratory Syndrome Coronavirus-2 (SARS-CoV-2) urges the scientist to identify the potential vaccine or drug to control the disease. SARS-CoV-2 with its single stranded RNA genome (length ~ 30 kb) is enveloped with active spike proteins. The genome is non-segmental with 5’-cap and 3’-poly tail and acts as a mRNA for the synthesis of replicase polyproteins. The replicase gene lying downstream to 5’-end encodes for non-structural protein, which in turn pose multiple functions ranging from envelope to nucleocapsid development. This study aims to identify the highly stable, effective and less toxic single strand RNA-based aptamers against non-structural protein 10 (NSP10). NSP10 is the significant activator of methyltransferase enzymes (NSP14 and NSP16) in SARS-CoV-2. Inhibiting the activation of methyltransferase leads to partial viral RNA capping or lack of capping, which makes the virus particles susceptible to host defence system. Results In this study, we focused on designing RNA aptamers through computational approach, docking of protein-aptamer followed by molecular dynamics simulation to perceive the binding stability of complex. Docking study reveals the high binding affinity of three aptamers namely RNA-053, 001, 010 to NSP10 with the HADDOCK score of − 88.5 ± 7.0, − 87.7 ± 11.5, − 86.1 ± 12 respectively. Molecular Dynamics suggests high conformational stability between the aptamer and the protein. Among the screened aptamers two aptamers maintained at least 3-4 intermolecular H-bonds throughout the simulation period. Conclusions The study identifies the potential aptamer candidate against less investigated but significant antiviral target i.e., NSP10/NSP16 interface complex.

Platelets in HIV: A Guardian of Host Defence or Transient Reservoir of the Virus?

The immune and inflammatory responses of platelets to human immunodeficiency virus 1 (HIV-1) and its envelope proteins are of great significance to both the treatment of the infection, and to the comorbidities related to systemic inflammation. Platelets can interact with the HIV-1 virus itself, or with viral membrane proteins, or with dysregulated inflammatory molecules in circulation, ensuing from HIV-1 infection. Platelets can facilitate the inhibition of HIV-1 infection via endogenously-produced inhibitors of HIV-1 replication, or the virus can temporarily hide from the immune system inside platelets, whereby platelets act as HIV-1 reservoirs. Platelets are therefore both guardians of the host defence system, and transient reservoirs of the virus. Such reservoirs may be of particular significance during combination antiretroviral therapy (cART) interruption, as it may drive viral persistence, and result in significant implications for treatment. Both HIV-1 envelope proteins and circulating inflammatory molecules can also initiate platelet complex formation with immune cells and erythrocytes. Complex formation cause platelet hypercoagulation and may lead to an increased thrombotic risk. Ultimately, HIV-1 infection can initiate platelet depletion and thrombocytopenia. Because of their relatively short lifespan, platelets are important signalling entities, and could be targeted more directly during HIV-1 infection and cART.

Genes of SARS-CoV-2 and emerging variants

The pandemic caused by severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) is distinctly different from outbreaks caused by other coronaviruses: SARS-CoV and Middle East respiratory syndrome coronavirus (MERS-CoV). The differences in the rapid transmission and severity of human coronaviruses are due to the genetic composition of the virus. SARS-CoV-2 contains genes encoding non-structural proteins (NSPs), structural proteins, and accessory proteins. The NSPs are mainly involved in replication of the virus within the host and inhibition of the host defence system. Structural proteins are involved in viral entry and attachment to host cells, preservation of the core virion and elicit the majority of the immune response. The functions of the accessory proteins are largely unknown. Most focus has been given to structural proteins, especially the spike protein as the strongest vaccine candidate. However, the recent emergence of spike variants and their ability to rapidly transmit and escape neutralisation by vaccine-induced antibodies has threatened the global community. Meanwhile, recent studies of accessory proteins reveal their importance in viral pathogenesis. Hence, proper understanding of the functions of all unknown viral proteins is crucial to devise alternate antiviral strategies.

Potential antibacterial effects of flaxseed and Nigella sativa extracts on Streptococcus pyogenes

Antibiotic resistance is a major global problem, associated with inadvertent drug usage. Herbal interventions are a therapeutic strategy that warrants greater research attention. Flaxseed and Nigella sativa are well recognized original super foods that have demonstrated potent anti-microbial and anti-biofilm activities. In the oral cavity, the bacterial population is a result of the dynamic relationship between pathogens and commensals Streptococcus pyogenes is an important global human Gram-positive pathogen that causes a wide variety of acute infections, it is highly virulent since it has the ability overcome the host defence system. This in vitro study aims to evaluate antimicrobial activity of flaxseed and Nigella sativa extract against S. pyogenes. Ethanolic extract of flaxseed and Nigella sativa extracts were prepared and the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) against S. pyogenes was estimated. The results of this study show that both extracts exhibited antibacterial activity against S. pyogenes. Present study demonstrated the bactericidal activity of both extracts which can be an adjunct to the future natural anti-bacterial therapy.

Emerging Complexity and the Need for Advanced Drug Delivery in Targeting Candida Species

Background: Candida species are the important etiologic agents for candidiasis, the most prevalent cause of opportunistic fungal infections. Candida invasion results in mucosal to systemic infections through immune dysfunction and helps in further invasion and proliferation at several sites in the host. The host defence system utilizes a wide array of the cells, proteins and chemical signals that are distributed in blood and tissues which further constitute the innate and adaptive immune system. The lack of antifungal agents and their limited therapeutic effects have led to high mortality and morbidity related to such infections. Methods: The necessary information collated on this review has been gathered from various literature published from 1995 to 2019. Results: This article sheds light on novel drug delivery approaches to target the immunological axis for several Candida species (C. albicans, C. glabrata, C. parapsilosis, C. tropicalis, C. krusei, C. rugose, C. hemulonii, etc.). Conclusion: It is clear that the novel drug delivery approaches include vaccines, adoptive transfer of primed immune cells, recombinant cytokines, therapeutic antibodies, and nanoparticles, which have immunomodulatory effects. Such advancements in targeting various underpinning mechanisms using the concept of novel drug delivery will provide a new dimension to the fungal infection clinic particularly due to Candida species with improved patient compliance and lesser side effects. This advancement in knowledge can also be extended to target various other similar microbial species and infections.

Structural basis of transcription inhibition by the DNA mimic protein Ocr of bacteriophage T7

Bacteriophage T7 infects Escherichia coli and evades the host defence system. The Ocr protein of T7 was shown to exist as a dimer mimicking DNA and to bind to host restriction enzymes, thus preventing the degradation of the viral genome by the host. Here we report that Ocr can also inhibit host transcription by directly binding to bacterial RNA polymerase (RNAP) and competing with the recruitment of RNAP by sigma factors. Using cryo electron microscopy, we determined the structures of Ocr bound to RNAP. The structures show that an Ocr dimer binds to RNAP in the cleft, where key regions of sigma bind and where DNA resides during transcription synthesis, thus providing a structural basis for the transcription inhibition. Our results reveal the versatility of Ocr in interfering with host systems and suggest possible strategies that could be exploited in adopting DNA mimicry as a basis for forming novel antibiotics.Impact statementDNA mimicry Ocr protein, a well-studied T7 phage protein that inhibits host restriction enzymes, can also inhibit host transcription through competing with sigma factors in binding to RNA polymerase.

Antimicrobial Proteins: Key Components of Innate Immunity

Antimicrobial peptides (AMPs) are the small ubiquitous self-defence products which are extensively distributed in plants. They can be classified into several groups, including thionins, defensins, snakins, lipid transfer proteins, glycine-rich proteins, cyclotides, and hevein-type proteins. AMPs are important mediators of an innate host defence system, with antimicrobial activities against a broad spectrum of microorganisms. AMPs can be extracted and isolated from different plants and plant organs such as stems, roots, seeds, flowers and leaves. They perform various physiological defensive mechanisms to eliminate viruses, bacteria, fungi and parasites, and so could be used as therapeutic and preservative agents.

Rapid evolution and conserved function of the piRNA pathway

Transposons are major genome constituents that can mobilize and trigger mutations, DNA breaks and chromosome rearrangements. Transposon silencing is particularly important in the germline, which is dedicated to transmission of the inherited genome. Piwi-interacting RNAs (piRNAs) guide a host defence system that transcriptionally and post-transcriptionally silences transposons during germline development. While germline control of transposons by the piRNA pathway is conserved, many piRNA pathway genes are evolving rapidly under positive selection, and the piRNA biogenesis machinery shows remarkable phylogenetic diversity. Conservation of core function combined with rapid gene evolution is characteristic of a host–pathogen arms race, suggesting that transposons and the piRNA pathway are engaged in an evolutionary tug of war that is driving divergence of the biogenesis machinery. Recent studies suggest that this process may produce biochemical incompatibilities that contribute to reproductive isolation and species divergence.

Molecular characterization and expression analysis of CSαβ defensin genes from the scorpion Mesobuthus martensii

Defensins are important components of innate host defence system against bacteria, fungi, parasites and viruses. Here, we predicted six potential defensin genes from the genome of the scorpion Mesobuthus martensii and then validated four genes from them via the combination of PCR and genomic sequence analysis. These four scorpion defensin genes share the same gene organization and structure of two exons and one phase-I intron with the GT-AG rule. Conserved motif and phylogenetic analysis showed that they belonged to the members of the invertebrate cysteine-stabilized α-helix/β-sheet motif defensin (CSαβ) defensin family. All these four CSαβ defensin genes have the expression feature of constitutive transcription (CON) by the whole scorpion infection model, promoter sequence analysis and dual luciferase assays. Further evolution and comparison analysis found that the invertebrate CSαβ defensin genes from most of arachnids and mollusks appear to share the expression pattern of CON, but those from insects and lower invertebrates (nematodes, annelids, cnidarians and sponges) seem to have identical inducible transcription (IND) after being challenged by microorganisms. Together, we identified four scorpion CSαβ defensin genes with the expression feature of CON, and characterized the diversified expression patterns of the invertebrate CSαβ defensin genes, which will shed insights into the evolution of the invertebrate CSαβ defensin genes and their expression patterns.

Diagnosis and management of shock in the ICU

Sepsis is triggered by an infection and treatment of sepsis requires timely identification of the patient, and rapid treatment with antibiotics, source control, and fluids. In the absence of a true biomarker for sepsis, the clinician needs to recognize which patients are at risk, as well as the common signs and symptoms of infection. The site of infection, the patient’s phenotype, and the location of the patient will help drive decisions about initial antibiotic therapy. Patients with sepsis should be treated to ensure adequate cardiac output and organ perfusion, which usually requires infusion of intravenous fluids. Crystalloid fluids are most frequently infused, and patients will often require large doses in the first 6–24 hours of treatment. In addition to haemodynamic and fluid support, some patients require infection source control. Many sepsis patients require additional supportive therapy with vasoactive agents, mechanical ventilation, renal replacement therapy, and nutritional therapy. The use of these supportive therapies allows for a patients host defence system to work in conjunction with antibiotics to fight off the infection. When using these supportive therapies, the clinician should attempt to minimize the complications of the therapies and the causative infection. Once a patient starts to clinically improve, it is essential that therapies that are no longer necessary are withdrawn. Patients who do not respond to initial therapy should be evaluated for either resistant organisms, persistent sources, or alternate diagnoses.