Video_1_Auto-Reactive Th17-Cells Trigger Obsessive-Compulsive-Disorder Like Behavior in Mice With Experimental Autoimmune Encephalomyelitis.MOV

Th17-lymphocytes are well known for their deleterious role in autoimmunity. But does the notoriety of this repertoire extend beyond autoimmunity? In the present study we employed experimental autoimmune encephalomyelitis as model system to study the role auto-reactive Th17 cells in neuropsychiatric disorders. The mice with experimental autoimmune encephalomyelitis exhibited exaggerated grooming activity. The observed behavioral anomaly resembled obsessive compulsive disorder (OCD) upon analysis of grooming microstructure, induced grooming, marble burying and nestlet shredding. The observed OCD like behavior was relieved upon Th17 cell depletion; alternatively, it could alone be induced by adoptive transfer of myelin oligodendrocyte glycoprotein (35-55) reactive Th17 in B6.Rag1−/− mice. The observed OCD like behavior was also alleviated upon treatment with a selective serotonin reuptake inhibitor, fluoxetine.</p

Image_1_Auto-Reactive Th17-Cells Trigger Obsessive-Compulsive-Disorder Like Behavior in Mice With Experimental Autoimmune Encephalomyelitis.pdf

Th17-lymphocytes are well known for their deleterious role in autoimmunity. But does the notoriety of this repertoire extend beyond autoimmunity? In the present study we employed experimental autoimmune encephalomyelitis as model system to study the role auto-reactive Th17 cells in neuropsychiatric disorders. The mice with experimental autoimmune encephalomyelitis exhibited exaggerated grooming activity. The observed behavioral anomaly resembled obsessive compulsive disorder (OCD) upon analysis of grooming microstructure, induced grooming, marble burying and nestlet shredding. The observed OCD like behavior was relieved upon Th17 cell depletion; alternatively, it could alone be induced by adoptive transfer of myelin oligodendrocyte glycoprotein (35-55) reactive Th17 in B6.Rag1−/− mice. The observed OCD like behavior was also alleviated upon treatment with a selective serotonin reuptake inhibitor, fluoxetine.</p

Trichloroisocyanuric acid mediated one-pot synthesis of 3,5-diarylisoxazoles from <i>α,β</i>-unsaturated ketones

A facile one-pot synthesis of 3,5-diarylisoxazoles from α,β-unsaturated ketones and hydroxylamine hydrochloride is reported. The reaction is efficiently promoted by trichloroisocyanuric acid (TCCA) to afford the desired products, mostly in high yields and in relatively short time. The mild nature of the synthesis and short reaction time are notable advantages of the developed protocol. This protocol is effective towards various substrates having different functionalities.</p

A convenient one-pot synthesis of N-fused 1,2,4-triazoles via oxidative cyclization using chromium (VI) oxide

A facile one-pot synthesis of N-fused 1,2,4-triazoles from heterocyclic hydrazines and aldehydes is reported. The reaction is efficiently promoted by chromium (VI) oxide to afford the desired products mostly in high yields and in relatively short time. The high yield of the products and short reaction time are notable advantages of the developed protocol. This protocol is effective toward various substrates having different functionalities.</p

Circular supply chain implementation performance measurement framework: a comparative case analysis

Circular supply chain (CSC) has gained traction amongst academicians, practitioners, and policymakers across the world due to its wide range of sustainable benefits to business organizations. CSC amalgamates the circular economy (CE) thinking into supply chain operations of industry and improves the three sustainability dimensions of the organizational performance. However, manufacturing organizations in developing economies are finding difficult to measure the impact of CSC adoption on organizational performance. Therefore, this research aims to explore the CSC performance measures and to develop a performance measurement framework for assessing the impact of CSC implementation on business organizational performance. This research proposes a modified balanced scorecard (BSC) based hybrid framework of Pythagorean fuzzy analytic hierarchy process (PF-AHP) and Pythagorean fuzzy weighted aggregated sum product assessment (PF-WASPAS) methods. The effectiveness of the proposed framework is validated through an empirical case study of an Indian manufacturing company. Further, the proposed framework is tested with other three Indian manufacturing companies and their results are compared with the case company. The finding reveals that the overall performance of empirical case company is 62.88% based on define set of performance measures and performance of other three companies are 64.51, 56.47, and 52.43%, respectively. The outcomes of this study shows that the proposed research framework is more reliable, consistent, and robust with circular perspectives and it also offers an effective way to measure and benchmark the impact of CSC adoption on organizational performance. This research contributes to the knowledge of CSC management for achieving sustainability in the business environment.</p

DataSheet_1_TLR4 activation by lysozyme induces pain without inflammation.pdf

Mostly, pain has been studied in association with inflammation, until recent studies which indicate that during bacterial infections, pain mechanisms could be independent of the inflammation. Chronic pain can sustain long after the healing from the injury, even in the absence of any visible inflammation. However, the mechanism behind this is not known. We tested inflammation in lysozyme-injected mice foot paw. Interestingly, we observed no inflammation in mice foot paw. Yet, lysozyme injections induced pain in these mice. Lysozyme induces pain in a TLR4-dependent manner and TLR4 activation by its ligands such as LPS leads to inflammatory response. We compared the intracellular signaling of MyD88 and TRIF pathways upon TLR4 activation by lysozyme and LPS to understand the underlying mechanism behind the absence of an inflammatory response upon lysozyme treatment. We observed a TLR4 induced selective TRIF and not MyD88 pathway activation upon lysozyme treatment. This is unlike any other previously known endogenous TLR4 activators. A selective activation of TRIF pathway by lysozyme induces weak inflammatory cytokine response devoid of inflammation. However, lysozyme activates glutamate oxaloacetate transaminase-2 (GOT2) in neurons in a TRIF-dependent manner, resulting in enhanced glutamate response. We propose that this enhanced glutaminergic response could lead to neuronal activation resulting in pain sensation upon lysozyme injections. Collectively we identify that TLR4 activation by lysozyme can induce pain in absence of a significant inflammation. Also, unlike other known TLR4 endogenous activators, lysozyme does not activate MyD88 signaling. These findings uncover a mechanism of selective activation of TRIF pathway by TLR4. This selective TRIF activation induces pain with negligible inflammation, constituting a chronic pain homeostatic mechanism.</p

Identification of novel inhibitors of <i>Neisseria gonorrhoeae</i> MurI using homology modeling, structure-based pharmacophore, molecular docking, and molecular dynamics simulation-based approach

MurI is one of the most significant role players in the biosynthesis of the peptidoglycan layer in Neisseria gonorrhoeae (Ng). We attempted to highlight the structural and functional relationship between Ng-MurI and D-glutamate to design novel molecules targeting this interaction. The three-dimensional (3D) model of the protein was constructed by homology modeling and the quality and consistency of generated model were assessed. The binding site of the protein was identified by molecular docking studies and a pharmacophore was identified using the interactions of the control ligand. The structure-based pharmacophore model was validated and employed for high-throughput virtual screening and molecular docking to identify novel Ng-MurI inhibitors. Finally, the model was optimized by molecular dynamics (MD) simulations and the optimized model complex with the substrate glutamate and novel molecules facilitated us to confirm the stability of the protein-ligand docked complexes. The 100 ns MD simulations of the potential lead compounds with protein confirmed that the modeled complexes were stable. This study identifies novel potential compounds with good fitness and docking scores, which made the interactions of biological significance within the protein active site. Hence, the identified compounds may act as new leads to design and develop Ng-MurI inhibitors. Communicated by Ramaswamy H. Sarma</p

Structure-based drug discovery to identify SARS-CoV2 spike protein–ACE2 interaction inhibitors

After the emergence of the COVID-19 pandemic in late 2019, the severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) has undergone a dynamic evolution driven by the acquisition of genetic modifications, resulting in several variants that are further classified as variants of interest (VOIs), variants under monitoring (VUM) and variants of concern (VOC) by World Health Organization (WHO). Currently, there are five SARS-CoV-2 VOCs (Alpha, Beta, Delta, Gamma and Omicron), two VOIs (Lambda and Mu) and several other VOIs that have been reported globally. In this study, we report a natural compound, Curcumin, as the potential inhibitor to the interactions between receptor binding domain (RBD(S1)) and human angiotensin-converting enzyme 2 (hACE2) domains and showcased its inhibitory potential for the Delta and Omicron variants through a computational approach by implementing state of the art methods. The study for the first time revealed a higher efficiency of Curcumin, especially for hindering the interaction between RBD(S1) and hACE-2 domains of Delta and Omicron variants as compared to other lead compounds. We investigated that the mutations in the RBD(S1) of VOC especially Delta and Omicron variants affect its structure compared to that of the wild type and other variants and therefore altered its binding to the hACE2 receptor. Molecular docking and molecular dynamics (MD) simulation analyses substantially supported the findings in terms of the stability of the docked complexes. This study offers compelling evidence, warranting a more in-depth exploration into the impact of these alterations on the binding of identified drug molecules with the Spike protein. Further investigation into their potential therapeutic effects in vivo is highly recommended. Communicated by Ramaswamy H. Sarma</p

Identification of thrombin inhibiting antithrombin-III like protein from <i>Punica granatum</i> using <i>in silico</i> approach and in vitro validation of thrombin inhibition activity in crude protein

Thrombosis is characterized by the formation of clots in the blood vessels. Antithrombin-III deficiency in the blood causes thrombus formation. Supplementing antithrombin-III may serve as anticoagulant therapy. In the present studies, an antithrombin like Protein from Punica granatum has been identified and characterized using in silico approach. Based on sequence homology, an ALPP was selected depending upon its highest binding affinity of −41.28 kcal/mol with thrombin. Thrombin structure complexed with ALPP was docked with TAME using AutoDock Vina. No binding was observed for TAME at Ser195 of thrombin. MD simulation (50 ns) was performed to evaluate the flexibility and stability of docked complexes. In vitro assays with crude protein showed 78% thrombin inhibition at 5 µg and calculated IC50 value was 0.188 µg. The presence of thrombin inhibitors in crude protein was also confirmed by reverse zymography. Thus, it is very likely that the protein identified from P. granatum may act as thrombin inhibitor.</p

Immuno-electron microscopy of recombinant SpaFED-piliated lactococcal cells.

<p>Immunogold pilin protein-labeling and electron microscopic analysis of recombinant WT (GRS1189) and SpaF pilin-deleted (GRS1126) SpaFED-piliated lactococci (including GRS71 cells as a control) were done using the techniques described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0113922#s4" target="_blank">Materials and Methods</a>. GRS1189 (<b>A and inset</b>) and GRS71 (<b>B</b>) cells are single-labeled with anti-SpaD serum and protein A-10-nm gold particles. GRS1189 cells are double-labeled either with SpaD (10-nm; white arrowhead) and SpaF (15-nm; gray arrowhead) antisera (<b>C</b>) or with SpaD (10-nm; white arrowhead) and SpaE (15-nm; black arrowhead) antisera (<b>D</b>). GRS1226 cells are single-labeled with SpaD antiserum (10-nm) (<b>E</b>) as well as double-labeled either with SpaD (10-nm) and SpaF (15-nm) antisera (<b>F</b>) or with SpaD (10-nm; white arrowhead) and SpaE (15-nm; black arrowhead) antisera (<b>G</b>). A scale bar with dimensions is included in each panel.</p