Lipids play a crucial role in the entry and egress of viruses, regardless of whether they are naked or enveloped. Recent evidence shows that lipid involvement in viral infection goes much further. During replication, many viruses rearrange internal lipid membranes to create niches where they replicate and assemble. Because of the close connection between lipids and inflammation, the derangement of lipid metabolism also results in the production of inflammatory stimuli. Due to its pivotal function in the viral life cycle, lipid metabolism has become an area of intense research to understand how viruses seize lipids and to design antiviral drugs targeting lipid pathways. Palmitoylethanolamide (PEA) is a lipid-derived peroxisome proliferator-activated receptor-α (PPAR-α) agonist that also counteracts SARS-CoV-2 entry and its replication. Our work highlights for the first time the antiviral potency of PEA against SARS-CoV-2, exerting its activity by two different mechanisms. First, its binding to the SARS-CoV-2 S protein causes a drop in viral infection of ~70%. We show that this activity is specific for SARS-CoV-2, as it does not prevent infection by VSV or HSV-2, other enveloped viruses that use different glycoproteins and entry receptors to mediate their entry. Second, we show that in infected Huh-7 cells, treatment with PEA dismantles lipid droplets, preventing the usage of these vesicular bodies by SARS-CoV-2 as a source of energy and protection against innate cellular defenses. This is not surprising since PEA activates PPAR-α, a transcription factor that, once activated, generates a cascade of events that leads to the disruption of fatty acid droplets, thereby bringing about lipid droplet degradation through β-oxidation. In conclusion, the present work demonstrates a novel mechanism of action for PEA as a direct and indirect antiviral agent against SARS-CoV-2. This evidence reinforces the notion that treatment with this compound might significantly impact the course of COVID-19. Indeed, considering that the protective effects of PEA in COVID-19 are the current objectives of two clinical trials (NCT04619706 and NCT04568876) and given the relative lack of toxicity of PEA in humans, further preclinical and clinical tests will be needed to fully consider PEA as a promising adjuvant therapy in the current COVID-19 pandemic or against emerging RNA viruses that share the same route of replication as coronaviruses.

Present work demonstrates the fabrication of new and facile sandwich-type electrochemical immunosensor based on palladium nanoparticles (PdNPs), polyaniline (PANI) and fullerene-C-60 nanocomposite film modified glassy carbon electrode (PdNP@PANI-C-60/GCE) for ultrasensitive detection of Prostate-specific antigen (PSA) biomarker. PdNP@PANI-C-60 was electrochemically synthesized on GCE and used as an electroactive substrate. PdNP@PANI-C-60 was characterized by scanning electron microscopy (SEM), energy-dispersive X-ray spectroscopy (EDS), cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS). Primary antibody anti-PSA (Ab1) was covalently immobilized on PdNP@PANI-C-60/GCE using NHS/EDC linkers. In the presence of PSA antigen, horseradish peroxidase secondary antibody (HRP-Ab2) was brought into the surface of the electrode, developing stable amplified signals of H2O2 reduction. Under the optimal conditions, a linear curve for determination of PSA at the proposed immunosensor was 1.6x10(-4) ng.mL(-1) to 38 ng.mL(-1) with a limit of detection (LOD) of 1.95x10(-5) ng.mL(-1). The proposed immunosensor was successfully validated in serum and urine samples towards PSA detection with satisfactory and acceptable results.

The novel corona virus disease 2019 (SARS-CoV 2) pandemic outbreak was alarming. The binding of SARS-CoV (CoV) spike protein (S-Protein) Receptor Binding Domain (RBD) to Angiotensin converting enzyme 2 (ACE2) receptor initiates the entry of corona virus into the host cells leading to the infection. However, considering the mutations reported in the SARS-CoV 2 (nCoV), the structural changes and the binding interactions of the S-protein RBD of nCoV were not clear. The present study was designed to elucidate the structural changes, hot spot binding residues and their interactions between the nCoV S-protein RBD and ACE2 receptor through computational approaches. Based on the sequence alignment, a total of 58 residues were found mutated in nCoV S-protein RBD. These mutations led to the structural changes in the nCoV S-protein RBD 3d structure with 4 helices, 10 sheets and intermittent loops. The nCoV RBD was found binding to ACE2 receptor with 11 hydrogen bonds and 1 salt bridge. The major hot spot amino acids involved in the binding identified by interaction analysis after simulations includes Glu 35, Tyr 83, Asp 38, Lys 31, Glu 37, His 34 amino acid residues of ACE2 receptor and Gln 493, Gln 498, Asn 487, Tyr 505 and Lys 417 residues in nCoV S-protein RBD. Based on the hydrogen bonding, RMSD and RMSF, total and potential energies, the nCoV was found binding to ACE2 receptor with higher stability and rigidity. Concluding, the hotspots information will be useful in designing blockers for the nCoV spike protein RBD. [Formula: see text]Communicated by Ramaswamy H. Sarma.

Screening of Prostate-specific antigen (PSA) in human blood is the most common approach to diagnose prostate cancer. The joint application of biology and electrochemistry has shown a tremendous rise in research towards the development of electrochemical diagnostic tools for various diseases. The present study demonstrates the development of an effective immunosensing platform incorporating hydroquinone (HQ) immobilized, fullerene-C-60 and copper nanoparticles (CuNPs) composite film on glassy carbon electrode (HQ@CuNPs-reduced-fullerene-C-60/GCE) for the selective, quick and trace detection of PSA. In order to fabricate immunosensor sequential immobilization of primary antibody (Ab(1)), blocking agent (bovine serum albumin (BSA)), antigen (prostate-specific antigen (PSA)) and secondary antibody (Ab(2)) tagged with horseradish peroxide (HRP) was carried out on HQ@CuNPs-reduced-fullerene-C-60/GCE. Electrochemical characterization and the signal response of immunosensor were tested using cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS). Due to the synergetic effect of fullerene-C-60 and CuNPs, the novel nanocomposite film exhibited excellent catalytic activity towards hydrogen peroxide (H2O2) reduction for greatly amplified immunosensing signals. HQ@CuNPs-fullerene-C-60/GCE exhibited a well-defined redox peak and accelerated electrochemical reduction of H2O2 without any interference of dissolved oxygen and false-positive result in phosphate buffer solution (PBS) at pH 7.0. The parameters influencing the electrochemical response were optimized. Under the optimized conditions, wide linearity between PSA concentrations and current responses ranging from 0.005 ng/mL to 20 ng/mL with the lower detection limit of 0.002 ng/mL was obtained at the proposed immunosensor. The clinical applicability of the proposed immunosensor was successfully tested in serum and urine samples. Results revealed that the proposed immunosensor may create new boundaries in the identification of PSA in human blood samples. (c) 2020 Elsevier Ltd. All rights reserved.

Obesity prevalence continues to be a foremost health concern across the globe leading to the development of major health risk conditions like type II diabetes, hyperlipidemia, hypertension and even cancers. Because of the deprived drug-based management system, there is an urgent need for the development of new drugs aiming at satiety and appetite control targets. Among the reported satiety signaling targets, 5HT2C receptor plays a crucial role in decreasing appetite and has become a promising target for the development of anti-obesity drugs. Lorcaserin, a 5HT2C receptor agonist and the only drug available in the market, was designed based on the receptor mechanism of action. Due to limited drug options available and considering the adverse drug effects of Lorcaserin, the development of new drugs which are highly specific toward the 5HT2C target and with lesser side effects is essential. The present study is majorly focused on developing new 5HT2C agonists through computational approaches like screening, docking, and simulation using Phase, QikProp, Glide and Desmond applications of the Schrodinger suite. Screening protocols resulted in eight best hit molecules with affinity for the receptor and among them, five hits displayed binding affinity toward the conserved residue Asp 134 of the receptor. The stability of the five molecules in complex with the 5HT2C receptor was studied through molecular dynamic simulations. Three molecules, ZINC32123870, ZINC40312983 and ZINC32124535, maintained stable interactions with the Asp 134 residue throughout the 50 ns simulation run time. Further, due to the high sequence similarity seen among the receptors of 5HT2 family, the three potential hits were cross validated against other subtypes 5HT2A and 5HT2B of the 5HT2 family to determine the specificity of the molecules against the target. Among the three hits, ZINC32124535 was identified as the best potential hit based on the hydrogen bond interaction percentage with Asp residue [5HT2A (Asp 155:60%); 5HT2B (Asp155: No interaction); 5HT2C (Asp 134:86%)]. The ZINC32124535 molecule produced one salt bridge and hydrogen bond interactions with Asp 134, alike the known drug Lorcaserin. Based on the results, ZINC32124535 was identified as the best potential hit against the 5HT2C receptor.

Background and objectives: Nearly 20–30% of the world’s population suffers from allergic rhinitis, among them 15% are progressing to asthma conditions. Sorghum bicolor profilin (Sorb PF), one of the panallergens, was identified, but the allergen specificity is not yet characterized. Materials and Methods: To map the antigenic determinants responsible for IgE binding, the present study is focused on in silico modeling, simulation of Sorb PF and docking of the Sorb PF peptides (PF1-6) against IgG and IgE, followed by in vivo evaluation of the peptides for its allergenicity in mice. Results: Peptide PF3 and PF4 displayed high docking G-scores (−9.05) against IgE only. The mice sensitized with PF3 peptide showed increased levels of IL5, IL12, TNF-alpha, and GMCSF when compared to other peptides and controls, signifying a strong, Th2-based response. Concurrently, the Th1 pathway was inhibited by low levels of cytokine IL2, IFN-γ, and IL-10 justifying the role of PF3 in allergenic IgE response. Conclusions: Based on the results of overlapping peptides PF3 and PF4, the N-terminal part of the PF3 peptide (TGQALVI) plays a crucial role in allergenic response of Sorghum profilin.

Caspase a protease family member, have a vital role in cell death and inflammation process. Caspase-3, an effector caspase controls the regulation of apoptosis and has an anti apoptotic function. The mechanical significance of restoring apoptosis signaling to selectively target malignant cells is utilized to develop strong therapeutic strategies by the caspase family of mortality - induction molecules. Caspase-3 has currently no clear role in treatment for tumor progression and tumor sensitivity. The present study was aimed to screen caspase for potential inhibitors using computer aided docking methodologies. For this, zinc natural molecule database molecules were screened using e-pharmacophore and ADME protocols along with docking studies. Docking analysis selected two molecules, namely ZINC13341044 and ZINC13507846 with G-scores -5.27 and -6.19 respectively. These two potential hits are predicted as caspase inhibitors based on the results and can be further processed for in vitro validation.

Abstract In India, Sorghum plant allergenicity was reported to be approximately 54.9%. Sorghum bicolor Polcalcin (Sorb PC) was identified as the panallergen but the specificity of this allergen is yet to be characterized. The present study was aimed to characterize the antigenic determinants of Sorb PC that are responsible for eliciting the IgE response. In silico modeling, simulation studies and docking of Sorb PC peptides (PC1–11) against IgG and IgE followed by in vivo evaluation was adopted. Peptide docking studies revealed PC 6 with highest G-score −12.85 against IgE followed by PC-11, 5, 1 and 7 (−10.91) peptides. The mice sensitized with PC7 peptide showed interleukin (IL) 4 (IL-4), IL-5, IL-12, TNF-α and GMCSF levels increased when compared with other peptides and controls, signifying a strong T helper type 2 (Th2)-based response. In tandem, the T helper type 1 (Th1) pathway was inhibited by low levels of cytokine IL-2, interferon γ (IFN-γ) and increased IL-10 levels justifying the role of PC7 in allergic IgE response. Considering the above data of overlapping peptides of PC6 and PC7, N-terminal part of the PC7 peptide (DEVQRMM) is found to play a crucial role in Sorghum Polcalcin allergenic response.

Saccharomyces cerevisiae Ypr147cp was found localized to lipid droplets but the physiological role of Ypr147cp remains unknown. Sequence analysis of Ypr147cp revealed an α/β hydrolase domain along with the conserved GXSXG lipase motif. Recombinant Ypr147cp showed both triacylglycerol lipase and ester hydrolase activities. Knock out of YPR147C led to accumulation of TAG in ypr147cΔ when compared to wild type (WT). In addition, transmission electron microscopic analysis of ypr147cΔ cells revealed a greater number of lipid bodies, justifying the increase in TAG content, and the phenotype was rescued upon overexpression of YPR147C in ypr147cΔ. Moreover, the lipid profiling confirmed the accumulation of fatty acids derived from neutral and phospholipids in ypr147cΔ cells. Based on these results, Ypr147cp is identified as a lipid droplet associated triacylglycerol lipase along with an ester hydrolyzing capacity.

In this study, we have developed an electrochemical immunosensor for the detection of prostate-specific antigen (PSA) based on gold nanoparticles (AuNPs) and a chitosan (CHI) nanocomposite film coated on a screen printed electrode (SPE). The modified SPE was characterized using scanning electron microscopy (SEM), energy dispersive X-ray spectroscopy (EDX), cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS). The immunosensor was fabricated by sequential immobilization of anti-PSA primary antibody, PSA antigen and HRP-tagged secondary antibody on the surface of AuNPs/CHI/SPE. When this immunosensor was placed in 2.5mM H2O2, as HRP substrate, and 1mM methylene blue (MB), as redox mediator, greatly amplified immunosensing signals were observed. Immunosensing signals for the reduction of H2O2 were monitored in phosphate buffer (pH 7.0) by CV and square wave voltammetry (SWV). Under optimized conditions steady state current increased linearly with PSA concentration over the range 1-18ng/ml with a detection limit of 0.001ng/ml. The fabricated immunosensor demonstrated excellent sensitivity, stability and reproducibility and therefore was successfully applied to analyse PSA in biological samples. Finally, the results indicate that the proposed immunosensor has potential in clinical screening of cancer biomarkers.

Systematic illustration of electrode fabrication.

Ymr210w was identified as a MAG (Monoacylglycerol) lipase. The accumulation of the phospholipids in the ymr210wΔ was not clearly understood. It was expressed in S. cerevisiae using pYES2/CT vector and His-tag purified recombinant protein confirmed TAG lipase activity. To further evaluate the role of YMR210w, ester hydrolase activity was also confirmed with pNP-acetate, pNP-butyrate and pNP - palmitate. GC-MS lipid profiling of ymr210wΔ showed an increase in the 15:0 Pentadecanoic acid by 76% among the total lipids. Phospholipid, Erucic acid 22:1 (Δ13) showed 43% increase while steryl esters showed significant changes with 16:0 hexadecanoic acid augmentations by 80% and 18:0 Octadecanoic acid by 165% when compared to wild type (WT). Increase in the steryl ester and TAG content supports the accumulation of lipid bodies in ymr210wΔ strain when compared with WT cells.

Mycoremediation of low molecular weight polycyclic aromatic hydrocarbons (PAH) is well studied. This study highlights the degradation of high molecular weight PAH (HMW-PAHs): Benz (a) anthracene (BAA), Dibenz (a, h) anthracene (DBA), and Indeno pyrene (IND) by Aspergillus terricola var americanus. The metabolism of the HMW-PAHs by this fungus was investigated in liquid submerged culture added with 60 mg/L of each compound. Depletion of BAA, DBA, and IND was 94.80, 90.16, and 93.80%, respectively, after 10 days of culture. Fungal broth solvent extracts were analyzed for the presence of metabolites by gas chromatography mass spectrometry and the extracellular enzymes produced were also assayed. The coexistence of two metabolic pathways, cytochrome P450 monooxygenase and ligninolytic pathway were identified. A high laccase activity was observed in BAA cultures on day 4 when compared to other PAH's studied. In conclusion, A. terricola might be a promising fungus for bioremediation of HMW-PAH mixtures co-metabolically.

Alterations in lipid metabolism have been progressively documented as a characteristic property of cancer cells. Though, human ABHD2 gene was found to be highly expressed in breast and lung cancers, its biochemical functionality is yet uncharacterized. In the present study we report, human ABHD2 as triacylglycerol (TAG) lipase along with ester hydrolysing capacity. Sequence analysis of ABHD2 revealed the presence of conserved motifs G205XS207XG209 and H120XXXXD125. Phylogenetic analysis showed homology to known lipases, Drosophila melanogaster CG3488. To evaluate the biochemical role, recombinant ABHD2 was expressed in Saccharomyces cerevisiae using pYES2/CT vector and His-tag purified protein showed TAG lipase activity. Ester hydrolase activity was confirmed with pNP acetate, butyrate and palmitate substrates respectively. Further, the ABHD2 homology model was built and the modelled protein was analysed based on the RMSD and root mean square fluctuation (RMSF) of the 100 ns simulation trajectory. Docking the acetate, butyrate and palmitate ligands with the model confirmed covalent binding of ligands with the Ser207 of the GXSXG motif. The model was validated with a mutant ABHD2 developed with alanine in place of Ser207 and the docking studies revealed loss of interaction between selected ligands and the mutant protein active site. Based on the above results, human ABHD2 was identified as a novel TAG lipase and ester hydrolase.

ABSTRACT Objective: Obesity was considered as a serious health concern apart from the age group in today’s population globally. The percentage of obese people in the world’s population is increasing at a faster rate, and health issues arising due to obesity are gradually increasing. Our present insilico study was aimed to screen out natural molecules against the peroxisome proliferator-activated receptor (PPAR), especially alpha aids in triggering the obesity. Methods: Several targets for treating obesity were identified, and one among such promising target was PPAR. Using the insilico applications such as natural database was screened and the molecules were further evaluated based on their docking score parameter with the receptor. Results: The docking methodology suggested that two molecules zinc02091671 and zinc02137525 were found to reproduce the similar type of interactions such as that of the known inhibitor and crystal ligand. Conclusion: The reported two molecules were found to be promising agonists based on the computational studies and can be advanced the in vitro based evaluation. Keywords: Obesity, Peroxisome proliferator-activated receptor, e-pharmacophore, QikProp, Docking.

ABSTRACT Objective: Obesity was considered as a serious health concern apart from the age group in today’s population globally. The percentage of obese people in the world’s population is increasing at a faster rate, and health issues arising due to obesity are gradually increasing. Our present insilico study was aimed to screen out natural molecules against the peroxisome proliferator-activated receptor (PPAR), especially alpha aids in triggering the obesity. Methods: Several targets for treating obesity were identified, and one among such promising target was PPAR. Using the insilico applications such as natural database was screened and the molecules were further evaluated based on their docking score parameter with the receptor. Results: The docking methodology suggested that two molecules zinc02091671 and zinc02137525 were found to reproduce the similar type of interactions such as that of the known inhibitor and crystal ligand. Conclusion: The reported two molecules were found to be promising agonists based on the computational studies and can be advanced the in vitro based evaluation. Keywords: Obesity, Peroxisome proliferator-activated receptor, e-pharmacophore, QikProp, Docking.

Background/Objectives: Bioremediation of High Molecular Weight PAHs (HMW PAHs) with a combination of microorganisms was investigated. In the present study, the PAHs degradation capacity of a microbial consortium consisting of fungal, bacterial cultures was evaluated. Within 7 days, consortium showed 64% and 69% degradation of Benzo (a) pyrene (BaP) and Benzo (k) fluoranthene (BkF) each PAH of 30mg/L concentration respectively as the sole carbon source. The culture broth was analyzed for the presence of metabolites by gas Chromatography-Mass Spectrometry (GC-MS) and the activities of enzymes produced by the consortium were also assayed during the degradation process. Coexistence of three metabolic pathways, cytochrome P450 monooxygenase with the detection of epoxy derivatives leading to phenols, trans-dihydrodiols, cis-dihyrodiols leading to salycyclicacid and catechol by bacterial dioxygenases and via quinones of ligninolytic pathway were identified. Besides, Microbial consortia also demonstrated good biosurfactant production capacity, achieving a maximum emulsification activity of 61% (BaP and BkF) in the presence of PAHs as sole carbon source. These results indicate that this consortium can be used in the bioremediation of PAHs mixtures cometabolically in the contaminated environments.Keywords: Biosurfactant, Degradation Pathways, GC-MS Metabolites, PAHs, Microbial Consortium

Objective: Current drugs to Alzheimer’s disease (AD) are targeted to delay the breakdown of acetylcholine, thereby increasing the concentration of acetylcholine released into synaptic cleft and enhancing cholinergic neurotransmission. This paper deals with screening and identification of acetylcholinesterase (AChE) inhibitors in solvent extracts of Hyptis suaveolens (HS). Methods: In search of natural inhibitors of AChE, this study is focused on extract of HS, a member of Lamiaceae. 1:4 ratio of methanolic extract is prepared with shade dried areal parts of HS plant. The extract was assayed by Ellman’s method for inhibition activity and then purified using ammonium sulfate precipitation and chromatography techniques. Gas chromatography-mass spectrometry (GC-MS) identified compounds were analyzed by docking studies. Results: Methanolic extract showed maximum percentage inhibition of 75.00±4.30 (2.1 mg/ml) with an IC value of 1.020±0.026 mg/ml. However, saturated ammonium sulfate precipitation of methanolic extract and further fractionation by gel permeation chromatography showed 86.00±1.30% AChE inhibition (AChEI) activity. Reverse phase high-performance liquid chromatography (RP-HPLC) fraction (retention time [RT] 5.170) showed significant inhibition when compared to the other peak (RT 6.643). RP-HPLC fraction (RT 5.170) with significant inhibition was identified as Eugenol by GC-MS analysis. In silico analysis of all the GC-MS identified molecules revealed Eugenol as possessing preeminent absorption, distribution, metabolism, elimination properties and a glide (docking) score of −9.14 kcal/mole with AChE enzyme of pacific electric ray (Torpedo californica - TcAChE) (PDB ID: 1EVE). Conclusion: Screening, purification and identification, and identification of diverse phytochemicals of the HS plant, as potent source of AChEI. Keywords: Acetylcholinesterase, Acetylcholnesterase inhibitor, Hyptis suaveolens, Alzheimer’s disease, Dementia. 50

In India, 20-30% of the human population suffer from allergic rhinitis and 15% of them develop asthma. Plant pollens are one of the causative aeroallergens and are mixture of a number of molecules including major and minor allergens (Panallergens). Profilin and polcalcin are the known pollen specific panallergens. Allergenicity of the Sorghum plant in Andhra Pradesh was found to be 54.9%. But the allergens responsible have not been characterized well. This study highlights identification and molecular characterization of Sorghum bicolor profilin (Sorb PF) and S. bicolor polcalcin (Sorb PC) allergen genes based on homology. The coding sequences of the two genes were PCR amplified from the cDNA constructed from Sorghum pollen total RNA. The gene sequences were deposited in NCBI, KC427126 and KC427125. Recombinantly expressed histidine tag (His-tag) purified Sorghum polcalcin and profilin confirmed 9 and 14 kDa proteins, respectively. Based on multiple sequence alignment and phylogenetic analysis, Sorghum polcalcin and profilin were found to be closely related with Cynodon dactylon, Phleum pratense and Oryza sativa grass species. In silico Algpred based screening of SorbPF and SorbPC showed an allergenicity score of 1.149 and 0.879, respectively. The structure of two Ef-hand sequences (DTNGDGKISLSEL and DTDGDGFIDFNEF) of SorbPC showed an exact match with Phlp7. It is concluded that Sorghum recombinant profilin and polcalcin proteins can be of potential use in developing diagnostic kits for allergenicity to Sorghum pollen grains.

Increase in obesity rates and obesity associated health issues became one of the greatest health concerns in the present world population. With alarming increase in obese percentage there is a need to design new drugs related to the obesity targets. Among the various targets linked to obesity, pancreatic lipase was one of the promising targets for obesity treatment. Using the in silico methods like structure based virtual screening, QikProp, docking studies and binding energy calculations three molecules namely zinc85531017, zinc95919096 and zinc33963788 from the natural database were reported as the potential inhibitors for the pancreatic lipase. Among them zinc95919096 presented all the interactions matching to both standard and crystal ligand and hence it can be further proceeded to drug discovery process.