Challenges in the diagnosis and management of autoimmune hemolytic anemia: A case-based approach. Experience from a tertiary care hospital in the Haryana region.

BACKGROUND: Autoimmune hemolytic anemia (AIHA) is a rare immune disorder which occurs when antibodies are directed against self red blood cells (RBCs) leading to hemolysis. AIHA is widely classified as warm autoimmune hemolytic anemia, cold agglutinin syndrome, mixed AIHA, paroxysmal cold hemoglobinuria and rarely drug induced AIHA. The pathogenesis of AIHA is complex interplay between genetic predisposition, immune dysregulation and enviornmental triggers. A direct antiglobulin test can be used to assess the immunological origin of the hemolysis in order to diagnose AIHA after identifying laboratory and clinical symptoms of hemolysis. OBJECTIVE: The objective is to understand underlying mechanism in AIHAs, and usage of targeted therapies to modulate specific components of the immune response. MATERIALS AND METHODS: We are hereby presenting a case series of 11 clinically suspected cases of AIHA in collaboration with their clinical features, immuno-hematological and other laboratory parameters, Flow cytometric analysis of lymphocyte subset in relevant cases, underlying etiology as well as serological subtype are also included. RESULTS: Majority of the patients were categorized as secondary AIHA (7/11, 63.63%). Out of 11 cases 7 were serologically subtyped as warm AIHA (7/11, 63.63%) ,2 cases were DaaT negative AIHA (2/11;18.18%), 2 cases were characterized as mixed AIHA subtype (2/11, 18.18%). CONCLUSION: Accurate subtyping of AIHA requires a systematic immunohematological approach coupled with comprehensive evaluations of clinical, hematological, and biochemical parameters.

Toxic effects of aniline in liver, gills and kidney of freshwater fish Channa punctatus after acute exposure.

Aniline (C6H5NH2) is one of the hazardous aromatic amine where an amino group -NH2) is connected to phenyl ring (C6H5). Based on the evaluation of the 96-hour LC50 of aniline, two sublethal concentrations (4.19 mg/l and 8.39 mg/l) were selected for acute exposure tests in freshwater fish Channa punctatus. The liver, gills and kidney of fish being the principal sites of xenobiotic material accumulation, respiration, biotransformation, and excretion are the focus of the present study. Throughout the exposure time, the comet assay revealed increased tail length and tail DNA percentage indicating maximum damage to liver, gills and kidney of treated group after 96 h. After acute exposure, there was a significant (p ≤ 0.05) increase in the enzymatic activity of glutathione-S-transferase (GST) and acetylcholinesterase (AChE), whereas decline in superoxide dismutase (SOD) and catalase (CAT) activity was observed. Meanwhile, levels of malondialdehyde (MDA) increased over the exposure period for both concentrations. After 96 h of exposure, degree of tissue change (DTC) was evaluated in liver, gill and kidney of aniline exposed fish. Additionally, light microscopy revealed multiple abnormalities in liver, gills and kidney of all the treated groups. Significant changes were observed in the levels of biochemical markers viz., glucose, triglyceride, cholesterol, aspartate transaminase, alanine transaminase and urea following a 96-hour exposure to aniline. Studies using ATR-FTIR and transmission electron microscopy (TEM) revealed changes in biomolecules and structural abnormalities in several tissues of the aniline-exposed groups in comparison to the control group respectively.

Evaluating efficacy of Pseudomonas sp. EN-4 to lower the toxic potential of 4-bromophenol and assessing its competency in simulated microcosm.

An indigenous bacterium Pseudomonas sp. EN-4 had been reported earlier for its ability to co-metabolise 4-bromophenol (4-BP), in presence of phenol (100 mg/L) as co-substrate. The present study was undertaken to validate the efficacy of biotransformation by comparing the toxicity profiles of untreated and EN-4 transformed samples of 4-BP, using both plant and animal model. The toxicity studies in Allium cepa (A. cepa) indicated to lowering of mitotic index (MI) from 12.77% (water) to 3.33% in A. cepa bulbs exposed to 4-BP + phenol, which reflects the cytotoxic nature of these compounds. However, the MI value significantly improves to 11.36% in its biologically treated counterpart, indicating normal cell growth. This was further supported by significant reduction in chromosomal aberrations in A. cepa root cells exposed to biologically treated samples of 4-BP as compared to untreated controls. The oxidative stress assessed by comparing the activity profiles of different marker enzymes showed that the activities of superoxide dismutase (SOD), ascorbate peroxidase (APX) and guaiacol peroxidase (GPX) were reduced by 56%, 72%, and 37% respectively, in EN-4 transformed samples of 4-BP + phenol compared to its untreated counterpart. Similar trends were evident in the comet assay of fish (Channa punctatus) blood cells exposed to untreated and biologically treated samples of 4-BP. The comparative studies showed significant reduction in tail length (72.70%) and % tail intensity (56.15%) in fish blood cells exposed to EN-4 treated 4-BP + phenol, compared to its untreated counterpart. The soil microcosm studies validated the competency of the EN-4 cells to establish and transform 4-BP in soil polluted with 4-BP (20 mg/kg) and 4-BP + phenol (20 + 100 mg/kg). The isolate EN-4 achieved 98.08% transformation of 4-BP in non-sterile microcosm supplemented with phenol, indicating to potential of EN-4 cells to establish along with indigenous microflora.

Evaluation of the Recombinant Bacterial Chitinases as Anti-proliferative and Anti-migratory Agents for the Human Breast Cancer Cell Line, MCF-7.

Chitinases, a glycosyl hydrolase family 18 members, have a wide distribution in both prokaryotes and eukaryotes, including humans. Regardless of the absence of endogenous chitin polymer, various chitinases and chitinase-like proteins (CLPs) have been reported in mammals. However, several other carbohydrate polymers, such as hyaluronic acid and heparan sulfate, show structural similarities with chitin, which could be a potential target of chitinase and CLPs. Heparan sulfate is part of the integral membrane proteins and involves in cell adherence and migration. Hence, to demonstrate the effect of chitinase on cancer cell progression, we selected two chitinases from Serratia marcescens, ChiB and ChiC, which function as exo- and endo-chitinase, respectively. The ChiB and ChiC proteins were produced recombinantly by cloning chiB and chiC genes from Serratia marcescens. The cell viability of the Michigan Cancer Foundation-7 (MCF-7) cells was studied using different concentrations of the purified recombinant proteins. Cell viability assay was performed using 3-(4, 5-dimethyl thiazolyl-2)-2, 5-diphenyltetrazolium bromide and water-soluble tetrazolium salt, and the effect of ChiB and ChiC on cell proliferation was studied by clonogenic assay. The cell migration study was analysed by wound healing, transwell migration, and invasion assays. Cell cycle analysis of propidium iodide-stained cells and cell proliferation markers such as pERK1/2, pAKT, and SMP30 were also done. It was observed that both ChiB and ChiC were able to impede cell viability, cell migration, and invasion significantly. These observations and our in silico molecular docking analysis suggest that ChiC is a potential anticancer agent and is more efficient than ChiB. Since the ChiC is able to inhibit both cancer cell proliferation and migration, it could be a potential candidate for the treatment of metastatic cancer.

Evaluation of haematological, genotoxic, cytotoxic and ATR-FTIR alterations in blood cells of fish Channa punctatus after acute exposure of aniline.

Aniline (C6H5NH2) an important intermediate in the organic and fine chemical industry, is ubiquitously used worldwide. It is one of the important building block for manufacturing of 4,4-methylene diphenyl diisocyanate (MDI), accelerators in rubber processing, dyes, tattoo inks, photographic chemicals, antioxidants, corrosion inhibitors, pharmaceuticals and antiseptics. The current study evaluated 96 h LC50 of aniline and based on this, two sublethal concentrations (4.19 mg/l and 8.39 mg/l) were selected for acute exposure studies in freshwater food fish Channa punctatus. Erythrocytes of fish are nucleated hence they play an important role in physiology, immune system, protein signalling and haemostatic condition along with respiration. Blood samples were collected after 24, 48, 72, and 96 h of exposure to study haematological, cytotoxic and genotoxic effects of sublethal concentrations of aniline in C. punctatus. Symbolic elevation in time and dose dependent DNA damage was observed by comet assay as well as micronuclei assay revealing maximum damage after 96 h of exposure. After aniline exposure, scanning electron microscopy and ATR-FTIR studies showed anomalies in structure and alterations in biomolecules of RBCs of aniline exposed group as compared to control group respectively. Semi prep HPLC studies revealed bioaccumulation potential of aniline in higher concentration exposed group.

Synergistic and additive interactions of Shewanella sp., Pseudomonas sp. and Thauera sp. with chlorantraniliprole and emamectin benzoate for controlling Spodoptera litura (Fabricius).

The imprudent use of insecticides causes the development of resistance in insect pest populations, contamination of the environment, biological imbalance and human intoxication. The use of microbial pathogens combined with insecticides has been proposed as an alternative strategy for insect pest management. This IPM approach may offer effective ways to control pests, in addition to lowering the risk of chemical residues in the environment. Spodoptera litura (Fabricius) is a major pest of many crops like cotton, maize, tobacco, cauliflower, cabbage, and fodder crops globally. Here, we evaluated the combined effects of new chemistry insecticides (chlorantraniliprole and emamectin benzoate) and entomopathogenic bacterial strains, Shewanella sp. (SS4), Thauera sp. (M9) and Pseudomonas sp. (EN4) against S. litura larvae inducing additive and synergistic interactions under laboratory conditions. Both insecticides produced higher larval mortality when applied in combination with bacterial isolates having maximum mortality of 98 and 96% with LC50 of chlorantraniliprole and emamectin benzoate in combination with LC50 of Pseudomonas sp. (EN4) respectively. The lower concentration (LC20) of both insecticides also induced synergism when combined with the above bacterial isolates providing a valuable approach for the management of insect pests. The genotoxic effect of both the insecticides was also evaluated by conducting comet assays. The insecticide treatments induced significant DNA damage in larval hemocytes that further increased in combination treatments. Our results indicated that combined treatments could be a successful approach for managing S. litura while reducing the inappropriate overuse of insecticides.

An Evolutionarily Conserved Strategy for Ribosome Binding and Host Translation Inhibition by ß-coronavirus Non-structural Protein 1.

An important pathogenicity factor of SARS-CoV-2 and related coronaviruses is Non-structural protein 1 (Nsp1), which suppresses host gene expression and stunts antiviral signaling. SARS-CoV-2 Nsp1 binds the ribosome to inhibit translation through mRNA displacement and induces degradation of host mRNAs. Here we show that Nsp1-dependent host shutoff is conserved in diverse coronaviruses, but only Nsp1 from ß-Coronaviruses (ß-CoV) inhibits translation through ribosome binding. The C-terminal domain (CTD) of all ß-CoV Nsp1s confers high-affinity ribosome binding despite low sequence conservation. Modeling of interactions of four Nsp1s with the ribosome identified the few absolutely conserved amino acids that, together with an overall conservation in surface charge, form the ß-CoV Nsp1 ribosome-binding domain. Contrary to previous models, the Nsp1 ribosome-binding domain is an inefficient translation inhibitor. Instead, the Nsp1-CTD likely functions by recruiting Nsp1's N-terminal "effector" domain. Finally, we show that a cis-acting viral RNA element has co-evolved to fine-tune SARS-CoV-2 Nsp1 function, but does not provide similar protection against Nsp1 from related viruses. Together, our work provides new insight into the diversity and conservation of ribosome-dependent host-shutoff functions of Nsp1, knowledge that could aid future efforts in pharmacological targeting of Nsp1 from SARS-CoV-2 and related human-pathogenic ß-CoVs. Our study also exemplifies how comparing highly divergent Nsp1 variants can help to dissect the different modalities of this multi-functional viral protein.

An evolutionarily conserved strategy for ribosome binding and inhibition by ß-coronavirus non-structural protein 1.

An important pathogenicity factor of SARS-CoV-2 and related coronaviruses is Nsp1, which suppresses host gene expression and stunts antiviral signaling. SARS-CoV-2 Nsp1 binds the ribosome to inhibit translation through mRNA displacement and induces degradation of host mRNAs through an unknown mechanism. Here we show that Nsp1-dependent host shutoff is conserved in diverse coronaviruses, but only Nsp1 from ß-CoV inhibits translation through ribosome binding. The C-terminal domain of all ß-CoV Nsp1s confers high-affinity ribosome-binding despite low sequence conservation. Modeling of interactions of four Nsp1s to the ribosome identified few absolutely conserved amino acids that, together with an overall conservation in surface charge, form the ß-CoV Nsp1 ribosome-binding domain. Contrary to previous models, the Nsp1 ribosome-binding domain is an inefficient translation inhibitor. Instead, the Nsp1-CTD likely functions by recruiting Nsp1's N-terminal "effector" domain. Finally, we show that a viral cis -acting RNA element has co-evolved to fine-tune SARS-CoV-2 Nsp1 function, but does not provide similar protection against Nsp1 from related viruses. Together, our work provides new insight into the diversity and conservation of ribosome-dependent host-shutoff functions of Nsp1, knowledge that could aide future efforts in pharmacological targeting of Nsp1 from SARS-CoV-2, but also related human-pathogenic ß-coronaviruses. Our study also exemplifies how comparing highly divergent Nsp1 variants can help to dissect the different modalities of this multi-functional viral protein.

Larvicidal, growth inhibitory and biochemical effects of soil bacterium, Pseudomonas sp. EN4 against Spodoptera litura (Fab.) (Lepidoptera: Noctuidae).

BACKGROUND: Spodoptera litura (Fabricius) (Lepidoptera: Noctuidae) also known as tobacco caterpillar, is one of the most serious polyphagous pests that cause economic losses to a variety of commercially important agricultural crops. Over the past few years, many conventional insecticides have been used to control this pest. However, the indiscriminate use of these chemicals has led to development of insecticide resistant populations of S. litura in addition to harmful effects on environment. Due to these ill effects, the emphasis is being laid on alternative eco-friendly control measures. Microbial control is one of the important components of integrated pest management. Thus, in search for novel biocontrol agents, the current work was carried out with the aim to evaluate the insecticidal potential of soil bacteria against S. litura. RESULTS: Among the tested soil bacterial isolates (EN1, EN2, AA5, EN4 and R1), maximum mortality (74%) was exhibited by Pseudomonas sp. (EN4). The larval mortality rate increased in a dose-dependent manner. Bacterial infection also significantly delayed the larval development, reduced adult emergence, and induced morphological deformities in adults of S. litura. Adverse effects were also detected on various nutritional parameters. The infected larvae showed a significant decrease in relative growth and consumption rate as well as efficiency of conversion of ingested and digested food to biomass. Histopathological studies indicated damage to the midgut epithelial layer of larvae due to the consumption of bacteria treated diet. The infected larvae also showed a significantly decreased level of various digestive enzymes. Furthermore, exposure to Pseudomonas sp. also caused DNA damage in the hemocytes of S. litura larvae. CONCLUSION: Adverse effects of Pseudomonas sp. EN4 on various biological parameters of S. litura indicate that this soil bacterial strain may be used as an effective biocontrol agent against insect pests.

A rare case of complicated malaria presenting as warm autoimmune hemolytic anemia.

Autoimmune hemolytic anemia (AIHA) has been rarely reported worldwide or from India as the underlying cause of anemia in malaria. We hereby present a case of complicated Plasmodium falciparum malaria with concomitant warm AIHA in a 31-year-old male. Direct Antiglobulin Test (DAT) was positive and elution studies showed pan-agglutination reaction. Clinico-hematological and serological follow-up of the patient was done post artesunate treatment until day 9. We suggest that it is important to establish the immune basis of anemia in malaria patients for guiding the treatment plan for the clinicians and providing packed red blood cell transfusion if required.

Fumarate and nitrite reduction by Prevotella nigrescens and Prevotella buccae isolated from Chronic Periodontitis patients.

OBJECTIVE: This study is an investigation of anaerobic nitrite and fumarate reduction/respiration abilities of two characterised Prevotella species namely Prevotella nigrescens (SS6B) and Prevotella buccae (GS6B) isolated from the periodontal pockets of chronic periodontitis (ChP) patients. METHODS: Isolation and identification of the periodontal bacteria from 20 patients showing clinical symptoms of ChP. Characterisation of anaerobic nitrite and fumarate reduction was done in P. nigrescens (SS6B) and P. buccae (GS6B) using reduction assays, inhibition assays with use of specific inhibitors, growth assays and enzyme activity assays. Degenerate PCR was used to detect and amplify nitrite reductase (nrfA) and fumarate reductase (frdA) gene sequences in these Prevotella isolates. In addition, molecular and in silico analysis of the amplified anaerobic reductase gene sequences was performed using NCBI conserved domain analysis, Interpro database and MegaX. RESULTS: We provided experimental evidence for presence of active nitrite and fumarate reductase activities through enzyme activity, reduction, inhibitor and growth assays. Moreover, we were able to detect presence of 505 bps nrfA gene fragment and 400 bps frdA gene fragment in these Prevotella spp. These fragments show similarity to multiheme ammonia forming cytochrome c nitrite reductases and fumarate reductases flavoprotein subunit, respectively. CONCLUSION: Anaerobic nitrite and fumarate respiration abilities in P. nigrescens and P. buccae isolates appear to be important for detoxification process and growth, respectively.

Vathasura Kudineer, an Andrographis based polyherbal formulation exhibits immunomodulation and inhibits chikungunya virus (CHIKV) under invitro conditions.

ETHNOPHARMACOLOGICAL RELEVANCE: Chikungunya disease (CHIKD) is caused by the alphavirus, chikungunya virus (CHIKV) and is characterized by acute fever and joint inflammation; the inflammation continues even after clearance of the virus from the system, persisting for several months to years. Currently, there are no modern medicines/vaccines available for its treatment and use of over-the-counter anti-inflammatory generic medicines to relieve symptoms is generally practiced. In India, Indian traditional medicines hold a lot of promise to treat this infection and are routinely used during outbreaks. AIM OF THE STUDY: In the present study, we characterized the phytochemical and physicochemical properties of aqueous and ethanol extracts of the Vathasura Kudineer (VSK), a Andrographis based Siddha polyherbal formulation. Additionally, we evaluated its immunomodulatory and antiviral potential using an in vitro system. MATERIALS AND METHODS: Aqueous and ethanolic extracts of VSK were prepared and their physico and phytochemical properties were obtained by biochemical and biophysical assays, HPTLC and FTIR. The aqueous extracts of VSK and several of its ingredients were evaluated for their cytotoxicity in Vero cells and using the maximum non-toxic concentration (MNTC), were processed further for evaluating their ability to inhibit CHIKV infection in Vero cells. We performed the co-treatment assay with ethanol extract of VSK and several of its ingredients to assess the antiviral activity against chikungunya virus on Vero cells and through pre-treatment assay (anti-adhesive effect), co-incubation assay (virucidal effect) and post-treatment assay (post-entry effect) were evaluated. Further, we tested the aqueous extract of VSK along with some of its ingredients for their immunomodulatory properties. We performed antioxidant and anti-inflammatory assays using LPS-simulated RAW 264.7 cells. For antioxidant capacity of extracts, we performed extra-cellular ABTS radical scavenging activity and intra-cellular effects on ROS generation and SOD activity. We assessed the effect on most important inflammatory mediators like Nitric oxide (NO) and Prostaglandin E2 (PGE2) and pro-inflammatory cytokines like interleukin-1 beta (IL-1ß) and tumor necrosis factor alpha (TNFα). RESULTS: We provided the fingerprint of the phytochemicals of both ethanol and aqueous extracts of VSK that can be used for identification. We observed that ethanol extract was able to inhibit CHIKV infection at MNTC with 48 h of treatment on Vero cells. Its ingredient VSKI-As (Anethum sowa) found to be most effective to show virucidal effect while VSKI-Cs (Clerodendrum serratum) and VSKI-Pn (Pipper nigrum) found to be effective in post-entry effect. VSK was able to show ABTS radical scavenging activity, reduce ROS generation, inhibit the inflammatory mediators (NO and PGE2) and pro-inflammatory cytokines (IL-1ß and TNFα) production in LPS-stimulated RAW 264.7 cells. CONCLUSIONS: We provided the evidence that VSK has both immunomodulatory as well as antiviral potential. It shows virucidal as well as post-entry effects on chikungunya virus. VSK can inhibit pro-inflammatory cytokines, IL-1ß and TNFα production by suppressing the inflammatory mediators, NO and PGE2.

Type and screen method and Coombs crossmatch method for pretransfusion testing: A prospective comparative study in a tertiary care hospital in Delhi.

BACKGROUND: The aim of pretransfusion testing (PTT) is to prevent the immune-mediated hemolytic reaction by the transfusion of incompatible donor red cells. The methods of PTT have evolved over the years and a new method of type and screen (T and S) was introduced, in which only ABO grouping, Rh typing, and antibody screening would be carried out with omission of routine Coombs crossmatch. Although T and S is an accepted method for PTT in developed countries, only a few studies in literature have evaluated its efficacy in India. AIM AND OBJECTIVE: The aim of the study was to compare T and S method with conventional Coombs crossmatch method for PTT. MATERIALS AND METHODS: Two thousand and fifty samples were randomly selected from the samples received in blood bank for requisition of blood transfusion after taking informed consent. ABO blood grouping and Rh typing were performed for each recipient's sample. "T and S" and "Coombs crossmatch" were done simultaneously by two different persons without knowing the result of each test. A commercially available three cell panel was used for antibody screening, in which the recipient's plasma was reacted with red cells in the low ionic strength solution Coombs Gel card at 37°C by column agglutination technology. RESULTS: Antibody screening was positive in 29 (1.41%) patients and negative in 2021 (98.59%) patients. Out of 29 patients, 27 had alloantibodies and 2 had autoantibody. Most common alloantibody found in our study was anti-D. Other antibodies found were anti-K, anti-C + D, anti-E, anti-C, anti-c, anti-Jka, and anti-Mi (a). Crossmatch on first attempt was compatible in 2028 (98.93%) patients and incompatible in 22 (1.07%) patients. Out of 29 patients who were positive for antibody screening, crossmatch was incompatible in nine patients. Crossmatch was compatible in twenty patients, who had positive antibody screen. However, crossmatch compatibility in these patients, reflect either absence of corresponding antigen or antigen present in low dose (heterozygous) in donor blood. On the other hand, out of 22 patients incompatible on first crossmatch, antibody screening was negative in 13 patients and was positive in only nine patients. Hence, 13 patients with antibody would have been missed by antibody screen alone. Kappa statistics was used to compare the efficacy of "type and screening" and "Coombs crossmatch." It showed κ =0.445 (P < 0.001) implying moderate agreement between the two variables of "T and S" and "Coombs crossmatch." CONCLUSION: T and S is a scientifically better method but needs to be implemented with caution. We need to develop our own cell panels having adequate representation of indigenous antigen (including In, Mi (a), etc.,). Large-scale studies need to be done in India with indigenous screening cells to evaluate efficacy and safety of T and S method.

Plant derived antimicrobial peptides: Mechanism of target, isolation techniques, sources and pharmaceutical applications.

Antimicrobial resistance is a global health and development threat which is caused by the excess and prolonged usage of antimicrobial compounds in agriculture and pharmaceutical industries. Resistance of pathogenic microorganisms to the already existing drugs represent a serious risk to public health. Plant sources such as cereals, legumes, fruits and vegetables are potential substrates for the isolation of antimicrobial peptides (AMP) with broad spectrum antimicrobial activity against bacteria, fungi and viruses with novel immunomodulatory activities. Thus, in the quest of new antimicrobial agents, AMPs have recently gained interest. Therefore, AMP can be used in agriculture, pharmaceutical and food industries. This review focuses on various explored and unexplored plant based food sources of AMPs, their isolation techniques and antimicrobial mechanism of peptides. Therefore, the literature discussed in this review paper will prove beneficial the research purposes for agriculture, pharmaceutical and food industries. PRACTICAL APPLICATIONS: Isolation of antimicrobial peptides (AMPs) can be done on industrial scale. AMP isolated from food sources can be used in pharmaceutical and agriculture industries. AMP from natural sources mitigate the problem of antimicrobial resistance. AMP isolated from food products can be used as nutraceutical.

Prevotella: An insight into its characteristics and associated virulence factors.

Prevotella species, a gram-negative obligate anaerobe, is commonly associated with human infections such as dental caries and periodontitis, as well as other conditions such as chronic osteomyelitis, bite-related infections, rheumatoid arthritis and intestinal diseases like ulcerative colitis. This generally harmless commensal possesses virulence factors such as adhesins, hemolysins, secretion systems exopolysaccharide, LPS, proteases, quorum sensing molecules and antibiotic resistance to evolve into a well-adapted pathogen capable of causing successful infection and proliferation in the host tissue. This review describes several of these virulence factors and their advantage to Prevotella spp. in causing inflammatory diseases like periodontitis. In addition, using genome analysis of Prevotella reference strains, we examined other putative virulence determinants which can provide insights as biomarkers and be the targets for effective interventions in Prevotella related diseases like periodontitis.

Recent efforts for drug identification from phytochemicals against SARS-CoV-2: Exploration of the chemical space to identify druggable leads.

Nature, which remains a central drug discovery pool, is always looked upon to find a putative druggable lead. The natural products and phytochemical derived from plants are essential during a global health crisis. This class represents one of the most practical and promising approaches to decrease pandemic's intensity owing to their therapeutic potential. The present manuscript is therefore kept forth to give the researchers updated information on undergoing research in allied areas of natural product-based drug discovery, particularly for Covid-19 disease. The study briefly shreds evidence from in vitro and in silico researches done so far to find a lead molecule against Covid-19. Following this, we exhaustively explored the concept of chemical space and molecular similarity parameters for the drug discovery about the lead(s) generated from in silico-based studies. The comparison was drawn using FDA-approved anti-infective agents during 2015-2020 using key descriptors to evaluate druglike properties. The outcomes of results were further corroborated using Molecular Dynamics studies which suggested the outcomes in alignment with chemical space ranking. In a nutshell, current research work aims to provide a holistic strategic approach to drug design, keeping in view the identified phytochemicals against Covid-19.

Discovery of Potent, Selective, and State-Dependent NaV1.7 Inhibitors with Robust Oral Efficacy in Pain Models: Structure-Activity Relationship and Optimization of Chroman and Indane Aryl Sulfonamides.

Voltage-gated sodium channel NaV1.7 is a genetically validated target for pain. Identification of NaV1.7 inhibitors with all of the desired properties to develop as an oral therapeutic for pain has been a major challenge. Herein, we report systematic structure-activity relationship (SAR) studies carried out to identify novel sulfonamide derivatives as potent, selective, and state-dependent NaV1.7 inhibitors for pain. Scaffold hopping from benzoxazine to chroman and indane bicyclic system followed by thiazole replacement on sulfonamide led to identification of lead molecules with significant improvement in solubility, selectivity over NaV1.5, and CYP2C9 inhibition. The lead molecules 13, 29, 32, 43, and 51 showed a favorable pharmacokinetics (PK) profile across different species and robust efficacy in veratridine and formalin-induced inflammatory pain models in mice. Compound 51 also showed significant effects on the CCI-induced neuropathic pain model. The profile of 51 indicated that it has the potential for further evaluation as a therapeutic for pain.

Critical analysis of two cases of major crossmatch incompatibility in tertiary care hospital in Delhi.

Rh blood group is one of the most complexes of the human blood groups system. RHD gene encodes the D antigen, and the RHCE encodes the C, c, E, and e antigens. Out of these, Rh D antigen is the most immunogenic while e antigen is the least immunogenic. Rh antibodies are produced in Rh-negative individuals following sensitization which occurs either through pregnancy or during blood transfusion. We hereby report two cases of anti-e antibody, both presenting as major crossmatch incompatibility.

Quinoxaline derivatives disrupt the base stacking of hepatitis C virus-internal ribosome entry site RNA: reduce translation and replication.

RNA-biased small molecules with a monoquinoxaline core target the L-shaped structure of subdomain IIa of Hepatitis C virus internal ribosome entry site (IRES) RNA in proximity to the Mg2+ binding site. The binding event leads to the destacking of RNA bases, resulting in the inhibition of IRES-mediated translation and HCV RNA replication.

lncRNA HULC facilitates efficient loading of HCV-core protein onto lipid droplets and subsequent virus-particle release.

The cellular lipid pool plays a central role in hepatitis C virus (HCV) life cycle, from establishing infection to virus propagation. Here, we show that a liver abundant long noncoding RNA, highly upregulated in liver carcinoma (HULC), is upregulated during HCV infection and manipulates the lipid pool to favour virus life cycle. Interestingly, HULC was found to be crucial for the increase in number of lipid droplets in infected cells. This effect was attributed to the role of HULC in lipid biogenesis. Further, we demonstrated that HULC knockdown decreases the association of HCV-core protein with lipid droplets. This exhibited a direct consequence on the release of HCV particles. The role of HULC in HCV-particle release was further substantiated by additional knockdown and mutation experiments. Additionally, we found that increased level of HULC in HCV-infected cells was a result of Retinoid X Receptor Alpha (RXRA)-mediated transcription, which seemed to be aided by HCV-core protein. Taken together, the results identify a distinct role of long noncoding RNA HULC in lipid dynamics during HCV infection, which provides new insights into the complex process of HCV propagation and pathogenesis.