HPIP and RUFY3 are noncanonical guanine nucleotide exchange factors of Rab5 to regulate endocytosis-coupled focal adhesion turnover.

While the role of endocytosis in focal adhesion turnover-coupled cell migration has been established in addition to its conventional role in cellular functions, the molecular regulators and precise molecular mechanisms that underlie this process remain largely unknown. In this study, we report that proto-oncoprotein hematopoietic PBX-interacting protein (HPIP) localizes to focal adhesions as well as endosomal compartments along with RUN FYVE domain-containing protein 3 (RUFY3) and Rab5, an early endosomal protein. HPIP contains two coiled-coil domains (CC1 and CC2) that are necessary for its association with Rab5 and RUFY3 as CC domain double mutant, that is, mtHPIPΔCC1-2 failed to support it. Furthermore, we show that HPIP and RUFY3 activate Rab5 by serving as noncanonical guanine nucleotide exchange factors of Rab5. In support of this, either deletion of coiled-coil domains or silencing of HPIP or RUFY3 impairs Rab5 activation and Rab5-dependent cell migration. Mechanistic studies further revealed that loss of HPIP or RUFY3 expression severely impairs Rab5-mediated focal adhesion disassembly, FAK activation, fibronectin-associated-ß1 integrin trafficking, and thus cell migration. Together, this study underscores the importance of HPIP and RUFY3 as noncanonical guanine nucleotide exchange factors of Rab5 and in integrin trafficking and focal adhesion turnover, which implicates in cell migration.

Unlocking genetic insights: Evaluating wheat RILs for physiobiochemical traits under terminal heat stress conditions.

BACKGROUND: The increasing impacts of heat stress on wheat production due to climate change has entailed the development of heat-resilient crop varieties. To address this, two hundred recombinant inbred lines (RILs) derived from a cross between WH711/WH1021 were evaluated in a randomized block design (RBD) with two replications at CCSHAU, Hisar, during 2018-19 under heat stress and non-stress conditions. Heat stress was induced by altering the date of sowing so that the grain filling stage coincide with heat stress. RESULTS: Heat stress adversely affects RILs performance, as illustrated by alterations in phenotypic traits. Highest coefficients of variations were recorded for TAA, CTD 1, WUE, CTD 2, Cc and A under non-stress and heat stress conditions whereas gs, WUEi and GY under non-stress and SPAD 1, SPAD 2, GY and NDVI 2 under heat-stress conditions recorded moderate estimates of coefficient of variations. CTD 2, TAA, E, WUE and A displayed a significant occurrence of both high heritability and substantial genetic advance under non-stress. Similarly, CTD 2, NDVI 2, A, WUEi, SPAD 2, gs, E, Ci, MDA and WUE exhibited high heritability with high genetic advance under heat-stress conditions. CONCLUSIONS: Complementary and duplicate types of interactions with number of controlling genes were observed for different parameters depending on the traits and environments. RILs 41, 42, 59, 74, 75, 180 and 194 were categorized as heat tolerant RILs. Selection preferably for NDVI 1, RWC, TAA, A, E and WUEi to accumulate heat tolerance favorable alleles in the selected RILs is suggested for development of heat resilient genotypes for sustainable crop improvement. The results showed that traits such as such as NDVI, RWC, TAA, A, E, and WUEi, can be effective for developing heat-resilient wheat genotypes and ensuring sustainable crop improvement.

The Autophagy Receptor TAX1BP1 (T6BP) improves antigen presentation by MHC-II molecules.

CD4+ T lymphocytes play a major role in the establishment and maintenance of immunity. They are activated by antigenic peptides derived from extracellular or newly synthesized (endogenous) proteins presented by the MHC-II molecules. The pathways leading to endogenous MHC-II presentation remain poorly characterized. We demonstrate here that the autophagy receptor, T6BP, influences both autophagy-dependent and -independent endogenous presentation of HIV- and HCMV-derived peptides. By studying the immunopeptidome of MHC-II molecules, we show that T6BP affects both the quantity and quality of peptides presented. T6BP silencing induces the mislocalization of the MHC-II-loading compartments and rapid degradation of the invariant chain (CD74) without altering the expression and internalization kinetics of MHC-II molecules. Defining the interactome of T6BP, we identify calnexin as a T6BP partner. We show that the calnexin cytosolic tail is required for this interaction. Remarkably, calnexin silencing replicates the functional consequences of T6BP silencing: decreased CD4+ T cell activation and exacerbated CD74 degradation. Altogether, we unravel T6BP as a key player of the MHC-II-restricted endogenous presentation pathway, and we propose one potential mechanism of action.

The long noncoding RNA (LINC-RBE) expression in testicular cells is associated with aging of the rat.

Long noncoding RNAs (lncRNAs) are important regulatory biomolecules responsible for many cellular processes. The aging of mammals is manifested by a slow and gradual decline of physiological functions after adulthood, progressively resulting in age-related diseases. Testis comprises different cell-types with defined functions for producing haploid gametes and androgens in males, contributing gene-pool to the next generation with genetic variations to species for evolutionary advantage. The LINC-RBE (long intergenic noncoding-rat brain expressed) RNA showed highest expression in the Leydig cells, responsible for steroidogenesis and production of testosterone; higher expression in primary spermatocytes (pachytene cells), responsible for generation of haploid gametes and high expression in Sertoli cells, the nursing cells of the testes. Testes of immature (4-weeks), adult (16- and 44-weeks), and nearly-old (70-weeks) rats showed low, high, and again low levels of expression, respectively. This along with the nuclear-cytoplasmic localization of LINC-RBE RNA showed age-related expression and function. Thus, expression of LINC-RBE is involved in the molecular physiology of testes, especially Leydig cells, primary spermatocytes, and Sertoli cells. The decline in its expression correlates with diminishing reproductive function of the testes during aging of the rat.

Cell-type specific and differential expression of LINC-RSAS long noncoding RNA declines in the testes during ageing of the rat.

Long noncoding RNAs (lncRNAs) have emerged as major regulators of gene expression, chromatin structure, epigenetic changes, post-transcriptional processing of RNAs, translation of mRNAs into proteins as well as contributing to the process of ageing. Ageing is a universal, slow, progressive change in almost all physiological processes of organisms after attaining reproductive maturity and often associated with age-related diseases. Mammalian testes contain various cell-types, vast reservoir of transcriptome complexity, produce haploid male gametes for reproduction and testosterone for development and maintenance of male sexual characters as well as contribute genetic variation to the species. We report age-related decline in expression and cellular localization of Long intergenic noncoding repeat-rich sense-antisense (LINC-RSAS) RNA in the testes and its major cell-types such as primary spermatocytes, Leydig cells and Sertoli cells during ageing of the rat. LINC-RSAS expression in testes increased from immature (4-weeks) to adult (16- and 44-weeks) and declined from adult (44-weeks) to nearly-old (70-weeks) rats. Genomic DNA methylation in the testes showed a similar pattern. Cell-type specific higher expression of LINC-RSAS was observed in primary spermatocytes (pachytene cells), Leydig cells and Sertoli cells of testes of adult rats. Over-expression of LINC-RSAS in cultured human cell lines revealed its possible role in cell-cycle control and apoptosis. We propose that LINC-RSAS expression is involved in molecular physiology of primary spermatocytes, Leydig cells and Sertoli cells of adult testes and its decline is associated with diminishing function of testes during ageing of the rat.

Morphological and molecular characterization of two species of genus Ageratum.

BACKGROUND: The species of genus Ageratum (family Asteraceae) are distributed in various parts of the world. Ageratum conyzoides and A. houstonianum are the most commonly occurring species in India. These species are quite similar in their morphology thus creating a challenge in identification during the field survey and taxonomic validation. The accurate identification of the species is highly significant especially when those are of medicinal interest. To overcome the barriers in morphological based identification, DNA barcoding has been employed during the present investigation. METHODS AND RESULTS: Morphological and DNA barcodes matK and ITS genes, were employed to differentiate between Ageratum conyzoides and A. houstonianum. The obtained matK and ITS gene sequences were submitted to GenBank and BOLD system to obtain accession numbers. The DNA sequences were aligned with database sequences using BLAST and phylogenetic trees were constructed through neighbor-joining algorithm in MEGA 11 software. The distinguish features of A. conyzoides include ovate to elliptic-oblong leaves with a cuneate base and inflorescence heads forming domed to flat-topped clusters. However, A. houstonianum has triangular to ovate leaves with a cordate to truncate base, cymose clusters in the inflorescence and stipulate glandular involucre bracts. The matK gene has shown the highest identity percentages (100%) for A. houstonianum and 99.87% for A. conyzoides. The phylogenetic tree analysis has demonstrated a close association of A. conyzoides and A. houstonianum with their respective species, supported by bootstrap values in the matK and ITS trees. CONCLUSION: This study revealed that morphological and molecular data can be successfully utilized in the identification of A. conyzoides and A. houstonianum. The matK and ITS barcodes provide promising results in the identification of Ageratum species, with their phylogeny supporting classification within the family asteraceae.

Identification of superior haplotypes for flowering time in pigeonpea through candidate gene-based association study of a diverse minicore collection.

KEY MESSAGE: In current study candidate gene (261 genes) based association mapping on 144 pigeonpea accessions for flowering time and related traits and 29 MTAs producing eight superior haplotypes were identified. In the current study, we have conducted an association analysis for flowering-associated traits in a diverse pigeonpea mini-core collection comprising 144 accessions using the SNP data of 261 flowering-related genes. In total, 13,449 SNPs were detected in the current study, which ranged from 743 (ICP10228) to 1469 (ICP6668) among the individuals. The nucleotide diversity (0.28) and Watterson estimates (0.34) reflected substantial diversity, while Tajima's D (-0.70) indicated the abundance of rare alleles in the collection. A total of 29 marker trait associations (MTAs) were identified, among which 19 were unique to days to first flowering (DOF) and/or days to fifty percent flowering (DFF), 9 to plant height (PH), and 1 to determinate (Det) growth habit using 3 years of phenotypic data. Among these MTAs, six were common to DOF and/or DFF, and four were common to DOF/DFF along with the PH, reflecting their pleiotropic action. These 29 MTAs spanned 25 genes, among which 10 genes clustered in the protein-protein network analysis, indicating their concerted involvement in floral induction. Furthermore, we identified eight haplotypes, four of which regulate late flowering, while the remaining four regulate early flowering using the MTAs. Interestingly, haplotypes conferring late flowering (H001, H002, and H008) were found to be taller, while those involved in early flowering (H003) were shorter in height. The expression pattern of these genes, as inferred from the transcriptome data, also underpinned their involvement in floral induction. The haplotypes identified will be highly useful to the pigeonpea breeding community for haplotype-based breeding.

Relationship of emotional intelligence and capability of answering higher-order knowledge questions in physiology among first-year medical students.

Emotional intelligence (EI) has a positive correlation with the academic performance of medical students. However, why there is a positive correlation needs further exploration. We hypothesized that the capability of answering higher-order knowledge questions (HOQs) is higher in students with higher EI. Hence, we assessed the correlation between EI and the capability of medical students to answer HOQs in physiology. First-year undergraduate medical students (n = 124) from an Indian medical college were recruited as a convenient sample. EI was assessed by the Schutte Self-Report Emotional Intelligence Test (SSEIT), a 33-item self-administered validated questionnaire. A specially designed objective examination with 15 lower-order and 15 higher-order multiple-choice questions was conducted. The correlation between the examination score and the EI score was tested by Pearson's correlation coefficient. Data from 92 students (33 females and 59 males) with a mean age of 20.14 ± 1.87 yr were analyzed. Overall, students got a percentage of 53.37 ± 14.07 in the examination, with 24.46 ± 9.1 in HOQs and 28.91 ± 6.58 in lower-order knowledge questions (LOQs). They had a mean score of 109.58 ± 46.2 in SSEIT. The correlation coefficient of SSEIT score with total marks was r = 0.29 (P = 0.0037), with HOQs was r = 0.41 (P < 0.0001), and with LOQs was r = 0.14 (P = 0.19). Hence, there is a positive correlation between EI and the capability of medical students to answer HOQs in physiology. This study may be the foundation for further exploration of the capability of answering HOQs in other subjects.NEW & NOTEWORTHY This study assessed the correlation between emotional intelligence (EI) and the capability of medical students to answer higher-order knowledge questions (HOQs) in the specific context of physiology. The finding reveals one of the multifaceted dimensions of the relationship between EI and academic performance. This novel perspective opens the door to further investigations to explore the relationship in other subjects and other dimensions to understand why students with higher EI have higher academic performance.

Nutritional, Shelf-Life, and Sensory Evaluation of Cucumis callosus Based Optimized Syrup.

Cucumis callosus or "Choti kachri" is feral species of Cucurbitaceae family grown commonly in the semi-arid zones. The current study aimed to optimize and develop a syrup by utilizing C. callosus. For optimization of various combinations of C. callosus (CC) and Malus domestica (MD), various treatments were prepared namely T1 (100:00), T2 (75:25), T3 (50:50), T4 (00:100). The developed syrups were analyzed for nutritional and sensory analysis for a storage period of three months. The maximum and minimum range of different blends and storage period for various parameters were reported as TSS (oBx) (65.06 ± 0.23 to 67.17 ± 0.25), pH (4.67 ± 0.17 to 5.02 ± 0.65), acidity (%citric acid) (0.61 ± 0.02 to 0.98 ± 0.11), reducing sugars (%) (35.98 ± 0.12 to 44.12 ± 0.03), total sugars (%) (57.12 ± 0.03 to 65.51 ± 0.07), and non-reducing sugars (%) (14.88 ± 0.19 to 26.65 ± 0.11). All the blends varied non- significantly in terms of TSS, pH and acidity while significant (p ≥ 0.05) difference was noted in sugars when blended with 75:25 ratio of CC: MD. The data with respect to storage study reveals that the TSS, PH and acidity varied non-significantly while reducing sugars increased significantly. However, total, and non-reducing sugars decreased significantly with the increase of storage period. The prepared blends were subjected for sensory evaluation using 9-point hedonic scale for assessing color, consistency, taste, aroma, and overall acceptability with noted values as 8.0 ± 0.54 to 7.0 ± 0.44, 7.2 ± 0.54 to 8.0 ± 0.70, 7.0 ± 0.44 to 8.0 ± 0.70, 7.2 ± 0.44 to 8.0 ± 0.70, and 7.2 ± 0.44 to 8.0 ± 0.83. The study indicated that C. callosus is an acceptable source for development of syrup.

Meta-QTL and ortho analysis unravels the genetic architecture and key candidate genes for cold tolerance at seedling stage in rice.

Rice, a critical cereal crop, grapples with productivity challenges due to its inherent sensitivity to low temperatures, primarily during the seedling and booting stages. Recognizing the polygenic complexity of cold stress signaling in rice, a meta-analysis was undertaken, focusing on 20 physiological traits integral to cold tolerance. This initiative allowed the consolidation of genetic data from 242 QTLs into 58 meta-QTLs, thereby significantly constricting the genetic and physical intervals, with 84% of meta-QTLs (MQTLs) being reduced to less than 2 Mb. The list of 10,505 genes within these MQTLs, was further refined utilizing expression datasets to pinpoint 46 pivotal genes exhibiting noteworthy differential regulation during cold stress. The study underscored the presence of several TFs such as WRKY, NAC, CBF/DREB, MYB, and bHLH, known for their roles in cold stress response. Further, ortho-analysis involving maize, barley, and Arabidopsis identified OsWRKY71, among others, as a prospective candidate for enhancing cold tolerance in diverse crop plants. In conclusion, our study delineates the intricate genetic architecture underpinning cold tolerance in rice and propounds significant candidate genes, offering crucial insights for further research and breeding strategies focused on fortifying crops against cold stress, thereby bolstering global food resilience. Supplementary Information: The online version contains supplementary material available at 10.1007/s12298-024-01412-1.

Defective T-cell response to COVID-19 vaccination in acute myeloid leukaemia and myelodysplastic syndromes.

Limited data exist on COVID-19 vaccination efficacy in patients with acute myeloid leukemia and myelodysplasia with excess blasts (AML/MDS-EB2). We report results from a prospective study, PACE (Patients with AML and COVID-19 Epidemiology). 93 patients provided samples post-vaccine 2 or 3 (PV2, PV3). Antibodies against SARS-COV-2 spike antigen were detectable in all samples. Neutralization of the omicron variant was poorer than ancestral variants but improved PV3. In contrast, adequate T-cell reactivity to SARS-COV-2 spike protein was seen in only 16/47 (34%) patients PV2 and 23/52 (44%) PV3. Using regression models, disease response (not in CR/Cri), and increasing age predicted poor T cell response.

Meta-QTL analysis in wheat: progress, challenges and opportunities.

Wheat, an important cereal crop globally, faces major challenges due to increasing global population and changing climates. The production and productivity are challenged by several biotic and abiotic stresses. There is also a pressing demand to enhance grain yield and quality/nutrition to ensure global food and nutritional security. To address these multifaceted concerns, researchers have conducted numerous meta-QTL (MQTL) studies in wheat, resulting in the identification of candidate genes that govern these complex quantitative traits. MQTL analysis has successfully unraveled the complex genetic architecture of polygenic quantitative traits in wheat. Candidate genes associated with stress adaptation have been pinpointed for abiotic and biotic traits, facilitating targeted breeding efforts to enhance stress tolerance. Furthermore, high-confidence candidate genes (CGs) and flanking markers to MQTLs will help in marker-assisted breeding programs aimed at enhancing stress tolerance, yield, quality and nutrition. Functional analysis of these CGs can enhance our understanding of intricate trait-related genetics. The discovery of orthologous MQTLs shared between wheat and other crops sheds light on common evolutionary pathways governing these traits. Breeders can leverage the most promising MQTLs and CGs associated with multiple traits to develop superior next-generation wheat cultivars with improved trait performance. This review provides a comprehensive overview of MQTL analysis in wheat, highlighting progress, challenges, validation methods and future opportunities in wheat genetics and breeding, contributing to global food security and sustainable agriculture.

Gut brain regulation using psychobiotics for improved neuropsychological illness.

"Psychobiotics" are a novel class of probiotics that are beneficial to the health and functional efficiency of our brain and psychology. The main hold on command in ill conditions of the brain and psychology is overtaken by these psychobiotic bacteria (a dietary supplement) via the action/determined role of bacterial neurochemicals or neuroactive substances that are released by them in the intestinal epithelium after their ingestion. Although these psychobiotics flourish in the gut of the host consuming them, the effect is widely spread to the brain due to the communication between the gut and the brain via the bidirectional gut-brain axis. The nervous system involved in this directional process includes both the enteric nervous system and the central nervous system. With time, several corroborations have proved the effectiveness of psychobiotics in terms of mental illnesses and brain disorders. In the prevailing situation of the coronavirus pandemic, psychobiotics may serve as an aid because a majority of the population worldwide is already suffering from psychological issues due to changes in lifestyle and dietary habits, and in need of an immediate solution to cope with it. Moreover, the in silico approach is also vital for the development of biological relevance to neurosubstances.

Phytofabrication of Silver Nanoparticles Using Trigonella foenum-graceum L. Leaf and Evaluation of Its Antimicrobial and Antioxidant Activities.

Silver nanoparticles (AgNPs) were fabricated using Trigonella foenum-graceum L. leaf extract, belonging to the variety HM 425, as leaf extracts are a rich source of phytochemicals such as polyphenols, flavonoids, and sugars, which function as reducing, stabilizing, and capping agents in the reduction of silver ions to AgNPs. These phytochemicals were quantitatively determined in leaf extracts, and then, their ability to mediate AgNP biosynthesis was assessed. The optical, structural, and morphological properties of as-synthesized AgNPs were characterized using UV-visible spectroscopy, a particle size analyzer (PSA), FESEM (field emission scanning electron microscopy), HRTEM (high-resolution transmission electron microscopy), and FTIR (Fourier transform infrared spectroscopy). HRTEM analysis demonstrated the formation of spherically shaped AgNPs with a diameter of 4-22 nm. By using the well diffusion method, the antimicrobial potency of AgNPs and leaf extract was evaluated against microbial strains of Staphylococcus aureus, Xanthomonas spp., Macrophomina phaseolina, and Fusarium oxysporum. AgNPs showed significant antioxidant efficacy with IC50 = 426.25 µg/mL in comparison to leaf extract with IC50 = 432.50 µg/mL against 2,2-diphenyl-1-picrylhydrazyl (DPPH). The AgNPs (64.36 mg AAE/g) demonstrated greater total antioxidant capacity using the phosphomolybdneum assay compared to the aqueous leaf extract (55.61 mg AAE/g) at a concentration of 1100 µg/mL. Based on these findings, AgNPs may indeed be useful for biomedical applications and drug delivery systems in the future.

Biosynthesis of Silver Nanoparticles Utilizing Leaf Extract of Trigonella foenum-graecum L. for Catalytic Dyes Degradation and Colorimetric Sensing of Fe3+/Hg2.

The aqueous Trigonella foenum-graecum L. leaf extract belonging to variety HM 444 was used as reducing agent for silver nanoparticles (AgNPs) synthesis. UV-Visible spectroscopy, Particle size analyser (PSA), Field emission scanning electron microscopy coupled to energy dispersive X-ray spectroscopy (FESEM-EDX) and High-resolution transmission electron microscopy (HRTEM) were used to characterize AgNPs. Selected area electron diffraction (SAED) confirmed the formation of metallic Ag. Fourier Transform Infrared Spectroscopy (FTIR) was done to find out the possible phytochemicals responsible for stabilization and capping of the AgNPs. The produced AgNPs had an average particle size of 21 nm, were spherical in shape, and monodispersed. It showed catalytic degradation of Methylene blue (96.57%, 0.1665 ± 0.03 min-1), Methyl orange (71.45%, 0.1054 ± 0.002 min-1), and Rhodamine B (92.72%, 0.2004 ± 0.01 min-1). The produced AgNPs were excellent solid bio-based sensors because they were very sensitive to Hg2+ and Fe3+ metal ions with a detection limit of 11.17 µM and 195.24 µM, respectively. From the results obtained, it was suggested that aqueous leaf extract demonstrated a versatile and cost-effective method and should be utilized in future as green technology for the fabrication of nanoparticles.

Pharmacological and Therapeutic Potential of Cucumis callosus: a Novel Nutritional Powerhouse for the Management of Non-communicable Diseases.

Cucumis callosus (Kachri) is an under-exploited fruit of the Cucurbitaceae family, distributed majorly in the arid regions of India in the states of Haryana, Rajasthan, and Gujarat. The fruit is traditionally used by the native people at a small scale by home-level processing. It is a perennial herb that has been shown to possess therapeutic potential in certain disorders. In several studies, the antioxidant, anti-hyperlipidaemic, anti-diabetic, anti-cancerous, anti-microbial, and cardioprotective properties of Kachri have been reported. The fruit has a good nutritional value in terms of high percentages of protein, carbohydrates, essential fatty acids, phenols, and various phytochemicals. Also, gamma radiation treatment has been used on this crop to reduce total bacterial counts (TBC), ensuring safety from pathogens during the storage period of the fruit and its products. These facts lay down a foundation for the development of functional food formulations and nutraceuticals of medicinal value from this functionally rich crop. Processing of traditionally valuable arid region foods into functional foods and products can potentially increase the livelihood and nutritional security of people globally. Therefore, this review focuses on the therapeutic and pharmacological potentials of the Kachri fruit in the management of non-communicable diseases (NCDs) namely, diabetes, cancer, and hyperlipidemia. Graphical abstract of the review.

Comparison of intrathecal bupivacaine-fentanyl and bupivacaine-butorphanol combinations for joint replacement surgeries.

Background and Aims: The objective of the study was to compare duration of analgesia of fentanyl versus butorphanol as adjuvants to bupivacaine in spinal anesthesia. Material and Methods: A prospective, randomized, double-blinded study conducted in 80 patients of 18-75 years age group and American Society of Anesthesiologists Grades I and II undergoing joint replacement surgeries. A total of 40 patients in each Group A and Group B received 0.5% bupivacaine 3 ml with 25 mcg fentanyl and 25 mcg butorphanol respectively, in a total volume of 3.5 ml made with saline. Duration of analgesia, number of rescue analgesia, sensory, and motor block characteristics were compared between the two groups. Statistical analysis was done using t test and Chi-square test with SPSS 19.0 software. Results: Mean duration of analgesia was found more in Group B in comparison to Group A (P < 0.05). A number of doses of analgesic required postoperatively were more in Group A compared to Group B (P < 0.001). Time required for onset of sensory and motor block was comparable in both the groups. However, two segment regression of sensory block was slower in Group B compared to Group A (P < 0.05). Conclusion: We conclude that addition of butorphanol 25 µg as an adjuvant to 0.5% hyperbaric bupivacaine provided prolonged duration of analgesia compared to 25 mg fentanyl.

Japanese encephalitis virus capsid protein interacts with non-lipidated MAP1LC3 on replication membranes and lipid droplets.

Microtubule-associated protein 1 light chain 3 (MAP1LC3) is a protein with a well-defined function in autophagy, but still incompletely understood roles in several other autophagy-independent processess. Studies have shown MAP1LC3 is a host-dependency factor for the replication of several viruses. Japanese encephalitis virus (JEV), a neurotropic flavivirus, replicates on ER-derived membranes that are marked by autophagosome-negative non-lipidated MAP1LC3 (LC3-I). Depletion of LC3 exerts a profound inhibition on virus replication and egress. Here, we further characterize the role of LC3 in JEV replication, and through immunofluorescence and immunoprecipitation show that LC3-I interacts with the virus capsid protein in infected cells. This association was observed on capsid localized to both the replication complex and lipid droplets (LDs). JEV infection decreased the number of LDs per cell indicating a link between lipid metabolism and virus replication. This capsid-LC3 interaction was independent of the autophagy adaptor protein p62/Sequestosome 1 (SQSTM1). Further, no association of capsid was seen with the Gamma-aminobutyric acid receptor-associated protein family, suggesting that this interaction was specific for LC3. High-resolution protein-protein docking studies identified a putative LC3-interacting region in capsid, 56FTAL59, and other key residues that could mediate a direct interaction between the two proteins.

Interplay among Structural Stability, Plasticity, and Energetics Determined by Conformational Attuning of Flexible Loops in PD-1.

The dynamics and plasticity of the PD-1/PD-L1 axis are the bottlenecks for the discovery of small-molecule antagonists to perturb this interaction interface significantly. Understanding the process of this protein-protein interaction (PPI) is of fundamental biological interest in structure-based drug designing. Food and Drug Administration (FDA)-approved anti-PD-1 monoclonal antibodies (mAbs) are the first-in-class with distinct binding modes to access this axis clinically; however, their mechanistic aspects remain elusive. Here, we have unveiled the interactive interfaces with PD-L1 and mAbs to investigate the native plasticity of PD-1 at global (structural and dynamical) and local (residue side-chain orientations) levels. We found that the structural stability and coordinated Cα movements are increased in the presence of PD-1's binding partners. The rigorous analysis of these PPIs using computational biophysical approaches revealed PD-1's intrinsic plasticity, its concerted loops' movement (BC, FG, and CC'), distal side-chain motions, and the thermodynamic landscape, which are perturbed remarkably from its unbound to bound states. Based on intra-/inter-residues' contact networks and energetics, the hot-spots have been identified that were found to be essential to arrest the dynamical motions of PD-1 significantly for the rational design of therapeutic agents by mimicking the mAbs mechanism.

Duration of postoperative analgesia with Nalbuphine vs Butorphanol as an adjunct to spinal anesthesia for lower limb orthopedic surgeries: A randomized double-blind active control trial.

Background and Aims: Nalbuphine as well as butorphanol as adjuvant to intrathecal bupivacaine have been studied in comparison to bupivacaine alone. Both are kappa receptor agonist and have never been compared for its efficacy in terms of postoperative analgesia. The aim of this study was to evaluate duration of postoperative analgesia as well as intraoperative block characteristics using intrathecal nalbuphine hydrochloride (800 µg) or butorphanol (25 µg) as adjuvant to hyperbaric bupivacaine (12.5 mg) in lower limb fracture femur surgeries as compared to active control, that is, saline and bupivacaine. Material and Methods: This prospective, randomized, double-blind, active control study was conducted on 90 adult patients of either sex belonging to ASA grade I/II, aged 18-70 years, being operated for fracture femur surgeries in tertiary care hospital of North India. Patients were randomly divided into 3 groups (n = 30) Group A: received 0.5% hyperbaric bupivacaine 12.5 mg with 800 µg nalbuphine. Group B: Received 0.5% hyperbaric bupivacaine 12.5 mg with 25 µg butorphanol. Group C: Received 0.5% hyperbaric bupivacaine 12.5 mg with normal saline. Total volume injected was 3.0 ml. Duration of analgesia, mean VAS scores, requirement of rescue analgesia in 24 h along with intraoperative sensory or motor characteristics of block and hemodynamic parameters were studied. Statistical analysis was done using ANOVA with post-hoc Tukey test, Student's t-test and Chi-Square test. Results: Demographic profile was comparable among all the three groups. Mean duration of postoperative analgesia was 348.33 ± 66.96, 156.17 ± 43.9 and 110.36 ± 29.18 min in group A, B, and C, respectively (P = 0.006). Total doses of rescue analgesia were least in group A (32), followed by group B (42) and group C (64), respectively (P = 0.001). Group A had significantly earlier onset of sensory action (P = 0.03) as compared to group B and C. There was significant difference in sensory (P = 0.08) and motor duration (P = 0.04) among all the three groups. However, onset of motor block, haemodynamic profile and side effects were comparable among groups A, B, and C (P > 0.05). Conclusion: Addition of 800 µg nalbuphine and 25 µg butorphanol as adjuvant to intrathecal bupivacaine has better outcome as compared to active placebo group. But intrathecal nalbuphine was more effective compared to intrathecal butorphanol in terms of prolonging postoperative analgesia, reducing rescue analgesic doses and onset of sensory block. However, hemodynamic profile and side effects were comparable among all groups.