Diversity of culturable gut bacteria of diamondback moth, Plutella xylostella (Linnaeus) (Lepidoptera: Yponomeutidae) collected from different geographical regions of India.

Diamondback moth, Plutella xylostella is one of the important pests of cole crops, the larval stages cause damage to leaves from seedling stage to the harvest thus reducing the quality and quantity of the yield. The insect gut posses a large variety of microbial communities among which, the association of bacteria is the most spread and common. Due to variations in various agro-climatic factors, the insect often assumes the status of major pest. These geographical variations in insects influence various biological parameters including insecticide resistance due to diversity of microbes/bacteria. The diverse role of gut bacteria in insect fitness traits has now gained perspectives for biotechnological exploration. The present study was aimed to determine the diversity of larval gut bacteria of diamondback moth collected from five different geographical regions of India. The gut bacteria of this pest were found to be influenced by different geographical regions. A total of 14 larval gut bacterial isolates were obtained. Majority of these bacteria belong to Enterobacteriaceae followed by Yersiniaceae, Morganellaceae and Enterococcaceae. Phylogeny analysis of all the bacterial isolates collected from five different geographical regions of India revealed that all the isolated strains are resolved in well-defined clades with their closest type species.

Antibacterial potential of hive bees honey from Himachal Pradesh, India.

This paper presents a pioneer study on the microbial diversity and antibacterial potential of hive bees (Apis cerana and A. mellifera) honey collected from Himachal Pradesh. In total, 26 bacteria (14 from A. cerana and 12 from A. mellifera) but no fungal isolate were recovered. Bee species and locations comparison in terms of bacterial load (log CFU/g) revealed maximum loads of 3.74 and 3.99 in the honey from A. cerana and Mandi location, respectively. The most prevalent strains (HC3, HC5, HC6, HC8 and HM2) were identified (16S rRNA ribotyping) as Staphylococcus haemolyticus (MT742636), "Bacillus subtilis subsp. stecoris" (MT742637), Bacillus safensis subsp. safensis (MT742638), "Bacillus zanthoxyli" (MT742639) and Bacillus safensis subsp. safensis (MT938911). The apiary honey displayed good to excellent inhibitory activity against Escherichia coli ATCC1041 whereas, fair to good against Bacillus subtilis ATCC6633, Pseudomonas aeruginosa ATCC10662, Salmonella typhi NCTC786 and Klebsiella pneumoniae ATCC13883, highlighting its use as a therapeutic agent. Furthermore, it can be effective in minimizing numerous side effects associated with the consumption of synthetic drugs for treating bacterial infections thereby signifying the role of honey as a healthier substitute for synthetic drugs.

A molecular basis for the exquisite CD1d-restricted antigen specificity and functional responses of natural killer T cells.

Natural killer T (NKT) cells respond to a variety of CD1d-restricted antigens (Ags), although the basis for Ag discrimination by the NKT cell receptor (TCR) is unclear. Here we have described NKT TCR fine specificity against several closely related Ags, termed altered glycolipid ligands (AGLs), which differentially stimulate NKT cells. The structures of five ternary complexes all revealed similar docking. Acyl chain modifications did not affect the interaction, but reduced NKT cell proliferation, indicating an affect on Ag processing or presentation. Conversely, truncation of the phytosphingosine chain caused an induced fit mode of TCR binding that affected TCR affinity. Modifications in the glycosyl head group had a direct impact on the TCR interaction and associated cellular response, with ligand potency reflecting the t(1/2) life of the interaction. Accordingly, we have provided a molecular basis for understanding how modifications in AGLs can result in striking alterations in the cellular response of NKT cells.

T cell receptor CDR2 beta and CDR3 beta loops collaborate functionally to shape the iNKT cell repertoire.

Mouse type I natural killer T cell receptors (iNKT TCRs) use a single V alpha 14-J alpha 18 sequence and V beta s that are almost always V beta 8.2, V beta 7, or V beta 2, although the basis of this differential usage is unclear. We showed that the V beta bias occurred as a consequence of the CDR2 beta loops determining the affinity of the iNKT TCR for CD1d-glycolipids, thus controlling positive selection. Within a conserved iNKT-TCR-CD1d docking framework, these inherent V beta-CD1d affinities are further modulated by the hypervariable CDR3 beta loop, thereby defining a functional interplay between the two iNKT TCR CDR beta loops. These V beta biases revealed a broadly hierarchical response in which V beta 8.2 > V beta 7 > V beta 2 in the recognition of diverse CD1d ligands. This restriction of the iNKT TCR repertoire during thymic selection paradoxically ensures that each peripheral iNKT cell recognizes a similar spectrum of antigens.

Pt-catalyzed rearrangement of oxaspirohexanes to 3-methylenetetrahydrofurans: scope and mechanism.

A novel Pt-catalyzed rearrangement of oxaspirohexanes to 3-methylenetetrahydrofurans is reported. Mechanistic studies by (13)C-labeling experiments confirm oxidative addition of Pt(II) regioselectively to the least substituted carbon-carbon bond of the cyclopropane to form a platinacyclobutane intermediate. To our knowledge, this is the first alkoxy-substituted platinacyclobutane that has been observed spectroscopically. The scope and a proposed mechanism of this new Pt-catalyzed transformation are described.

Synthesis of a 2"-deoxy-ß-GalCer.

Structural studies of ternary complexes of CD1d/glycosyl ceramides/iNKT cells and CD1d/sulfatide/sulfatide reactive Type II NKT cells have shown how the polar moieties on the glycolipids interact with both the antigen presenting protein (CD1d) and the T cell receptors. However, these structures alone do not reveal the relative importance of these interactions. This study focuses on the synthesis of the previously unknown 2"-deoxy-ß-galactosyl ceramide 2. This glycolipid is also evaluated for its ability to stimulate iNKT cells and sulfatide-reactive Type II NKT cells.

Impact of capacity building and tele ECG based decision support on change in thrombolysis rate and inhospital and one year mortality in patients with STEMI, using hub and spoke model; multi-phasic intervention trial.

BACKGROUND: We report the impact of capacity building and teleconsultation on change in the thrombolysis rates and one-year mortality in patients with STEMI using a hub and the spoke model of STEMI care. METHODS: Twenty secondary care public hospitals were linked with a teaching hospital as a hub centre and the impact of the intervention on change in ischemic time, thrombolysis rates and all-cause in-hospital and one-year mortality was compared. RESULTS: 29 patients with STEMI were treated during pre-intervention from April 2020 to June 2020 and 255 patients during the post-intervention period from July 2020 to Oct 2021 in spoke centres. 245 patients were reported to a hub centre during the study period. The thrombolysis rate was significantly higher in the spoke centres after intervention (65.5%vs. 27.5 % p < 0.001) and was also significantly higher than in patients treated in a hub centre (65.5 % vs. 45.7 % p < 0.01). The in-hospital mortality was significantly lower in patients treated at spoke centres compared to those treated at the hub centre (7.8 % vs. 15.5 % < 0.003). The significant difference in mortality rate continued at one year (11.0 % vs.18.4 % p < 0.01). The median time from symptoms to thrombolytic therapy was significantly lower in STEMI patients treated in spoke centres compared to a hub centre (230 min vs. 356 min p < 0.001). CONCLUSION: The hub and spoke model of STEMI care is effective in increasing thrombolysis rate, and decreasing in-hospital and one-year mortality rate.

Current Status and Medicinal Prominence of Arnebia euchroma (Ratanjot): A Critically Endangered Plant of Trans-Himalayan Region.

Trans-Himalayan region has been a major component of the India's opulent medicinal plant heritage that encompasses numerous critically endangered plant species. Arnebia euchroma (Royle ex Benth.) Johnston (common name: Ratanjot), a Trans- Himalayan native, is amongst them, and it belongs to the family Boraginaceae. Ratanjot has long been used as a colourant in food and cosmetics besides a major ingredient of traditional remedies prescribed for curing mild constipation, dermatitis, frostbite, and eczema like health disorders. Though principally harvested for its roots, almost all the parts of this plant have been used in pharmaceutical products, food, dyes and beverages since prehistoric times. Its roots are a rich source of naphthoquinone pigment(s) mainly shikonin, acetylshikonin and deoxyshikonin, accountable for its medicinal value as antimicrobial, wound healing, anti-inflammatory, anticancer, and antioxidant agent(s). Considering the medicinal importance and critically endangered status of this taxon, the need of the hour is to conserve and propagate it for supplying sufficient raw materials for its commercial exploitation.

Lethal and sublethal effects of thiamethoxam, a neonicotinoid molecule, on colony performance of A. mellifera.

Among insect pollinators, honey bees, Apis mellifera (Hymenoptera: Apidae), are universally acknowledged, most important managed pollinators that also provide honey production. In recent years, neonicotinoids are widely used against a broad spectrum sucking pests. However, they also pose a major threat to the beekeeping industry. The present study aimed to quantify the impact of thiamethoxam, a second-generation, broad-spectrum neonicotinoid on foraging behavior, colony performance, and survival of Apis mellifera L. in mustard crop under semi-field (cage) and field conditions. Under semi-field conditions, the foraging activity of A. mellifera on mustard bloom reduced significantly on the 2nd day after spray of thiamethoxam as compared to pre-count and control. Significant decrease in brood area (7th to 21st day), nectar stores (7th to 28th day), and pollen stores (7th to 21st day) were also recorded after the spray. The bee mortality under semi-field conditions was significantly higher on the 1st and 2nd day after spray in comparison to control. Under field conditions, average bee activity remained statistically low up to the 12th day after spray on mustard bloom in comparison to pre-count and control. The effect of thiamethoxam under field conditions was less pronounced for bee mortality and colony parameters. Based on LD50, thiamethoxam was proved toxic to adults and larvae of A. mellifera.

Synthesis and evaluation of 3''- and 4''-deoxy and -fluoro analogs of the immunostimulatory glycolipid, KRN7000.

Four 3''- and 4''-deoxy and -fluorogalactosyl ceramides were synthesized, and their ability to stimulate iNKT cells, based on levels of IL-2 production, was assessed in three NKT cell receptor hybridomas. In two of the hybridomas, 1.2 and 2H4, all of the analogs were immunostimulatory, while in the 1.4 hybridoma only the 4''-fluoro analog led to the production of significant levels of IL-2.

Applications of the Tunable Diode Laser Absorption Spectroscopy: In-Process Estimation of Primary Drying Heterogeneity and Product Temperature During Lyophilization.

The aim of this research was to evaluate the impact of variability in ice sublimation rate (dm/dt) measurement and vial heat transfer coefficient (Kv) on product temperature prediction during the primary drying phase of lyophilization. The mathematical model used for primary drying uses dm/dt and Kv as inputs to predict product temperature. A second-generation tunable diode laser absorption spectroscopy (TDLAS)-based sensor was used to measure dm/dt. In addition, a new approach to calculate drying heterogeneity in a batch during primary drying is described. The TDLAS dm/dt measurements were found to be within 5%-10% of gravimetric measurement for laboratory- and pilot-scale lyophilizers. Intersupplier variability in Kv was high for the same "type" of vials, which can lead to erroneous product temperature prediction if "one value" of vial heat transfer coefficient is used for "all vial types" from different suppliers. Studies conducted in both a laboratory- and a pilot-scale lyophilizer showed TDLAS product temperature to be within ±1°C of average thermocouple temperature during primary drying. Using TDLAS data and calculations to estimate drying heterogeneity (number of vials undergoing primary drying), good agreement was obtained between theoretical and experimental results, demonstrating usefulness of the new approach.

A minimal binding footprint on CD1d-glycolipid is a basis for selection of the unique human NKT TCR.

Although it has been established how CD1 binds a variety of lipid antigens (Ag), data are only now emerging that show how alphabeta T cell receptors (TCRs) interact with CD1-Ag. Using the structure of the human semiinvariant NKT TCR-CD1d-alpha-galactosylceramide (alpha-GalCer) complex as a guide, we undertook an alanine scanning mutagenesis approach to define the energetic basis of this interaction between the NKT TCR and CD1d. Moreover, we explored how analogues of alpha-GalCer affected this interaction. The data revealed that an identical energetic footprint underpinned the human and mouse NKT TCR-CD1d-alpha-GalCer cross-reactivity. Some, but not all, of the contact residues within the Jalpha18-encoded invariant CDR3alpha loop and Vbeta11-encoded CDR2beta loop were critical for recognizing CD1d. The residues within the Valpha24-encoded CDR1alpha and CDR3alpha loops that contacted the glycolipid Ag played a smaller energetic role compared with the NKT TCR residues that contacted CD1d. Collectively, our data reveal that the region distant to the protruding Ag and directly above the F' pocket of CD1d was the principal factor in the interaction with the NKT TCR. Accordingly, although the structural footprint at the NKT TCR-CD1d-alpha-GalCer is small, the energetic footprint is smaller still, and reveals the minimal requirements for CD1d restriction.