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Background: Pregnancy is a unique physiological diabetogenic state characterised by increased insulin resistance that ensures adequate supply of nutrients to the developing fetus. The insulin sensitivity falls to upto 50 percent in the late pregnancy. Thus insulin resistance and the resultant hyperinsulinemia are the characteristics features that are evident in the normal pregnancy during third trimester. In Hypertensive disorders of pregnancy (HDP), there is exacerbation of the physiological insulin resistance that occurs in normal pregnancy resulting in increased fasting serum insulin level. Methods: This is a case control study conducted on 90 antenatal women, during the study period of one and half years (from December 2020 to June 2022) in IMS and SUM Hospital, Bhubaneswar. With informed written consent and after fulfilling the criterias, 60 normotensive patients were chosen as controls and 30 pregnant patients with hypertensive disorders of pregnancy were chosen as cases. After 8 hours of overnight fasting, 2ml of blood is drawn and processed by CMIA technology to detect fasting serum insulin levels. The mean fasting serum insulin levels were compared between the cases and the controls. Results: The mean fasting serum insulin level of controls was found to be 9.27 and the mean fasting serum insulin level of cases was found to be 15.01 which was higher than controls. This was found to be statistically significant with a P value of 0.000. Conclusions: Increased fasting serum insulin level is observed in women with HDP than normotensive pregnant women.

Terminalia chebula fruits proteins antidiabetic activities were studied in in vitro model. Using appropriate standard methods were used to quantify the phytochemicals present in Terminalia chebula fruits extract. The results revealed that, the extract rich with proteins when compared to other phyto chemicals. The DPPH model was used for the analysis of antioxidant activity of the crude proteins. The alpha amylase enzyme, alpha glucosidases enzyme inhibition activity and Glucose uptake in Yeast cells studies were done to check the antiglycemic activity. These results revealed that, the crude protein showed antidiabetic activity. This potential pharmacological activity of proteins might be due to the presence of proteins.

The investigation of microplastics have been carried out among the commercially important marine fishes at Pulicat lake waters. The gastrointestinal tract of fishes were isolated and experimented for further analysis to detect the presence of microplastics. The fishes viz., Nemipterus japonicus, Mugil cephalus, Scomberomorus guttatus, Terapon puta, Sardinella longiceps, Caranx ignobilis, Stolephorus indicus, Rastrelliger kanagurta, Eleuthronema tetradactylum, Alectis indica, Leiognathus sp., Trichiurus lepturus, Trachurus lathamni, Selaroides leptolepis and Terapon jarbua were examined in this study. The microplastics samples were confirmed with BrukerAlpha FTIR ATR spectrophotometer in order to confirm plastics functional groups. Out of the 16 plastic particles isolated, 8 particles were mesoplastics and the rest of them were microplastics. The FTIR-ATR peaks were confirmed C-H stretching and N- H stretching (polyamide), C-H stretching, O-H stretching or N-H stretching (poly lactic acid), CH group (polythene) and C-H stretching, C=O stretching and C-O stretching (polyester). The presence of microplastics in the fish from the sampling locations may be due to unmonitored dumping of wastes in the water. The presence of microplastics may affect the growth of fish which leads to poor commercial value. The future research should be aimed to evaluate the impact and the risk of microplastics to the fishes, environment as well as human beings and to create awareness among the people, as well as the fishermen.

The investigation of microplastics has been carried out among Indian Major Carps from three lakes of Coimbatore, India. The gastrointestinal tract of fishes were isolated and examined to detect the presence of microplastics. The fishes of Indian major carps viz., Catla catla, Labeo rohita and Cirrhinus mrigala were used in this study. Three lakes were selected namely Periyakulam Lake, Kurichi Lake and Singanallur Lake in Coimbatore district, India. The microplastics samples were confirmed with Bruker – Alpha FTIR ATR spectrophotometer in order to confirm plastics functional groups. The presence of microplastics in the fish from the sampling locations may be due to unmonitored dumping of wastes in the water. The presence of microplastics may affect the growth of fish which leads to poor commercial value. The future research should be aimed to evaluate the impact and the risk of microplastics to the fishes, environment as well as to human beings.

SUMMARYChromosomal barcoding and high-throughput sequencing were used to investigate the population dynamics of Streptococcus pneumoniae. During infant mouse colonization, >35-fold reduction in diversity and expansion of a single clonal lineage was observed within 1 day post-inoculation. This loss of diversity was not due to immune factors, host microbiota or exclusively because of genetic drift. Rather, it required the expression of blp bacteriocins induced by the BlpC-quorum sensing pheromone. This points towards the role of intra-strain competition whereby the subpopulation reaching a quorum eliminates others that have yet to activate the blp locus. We show that this loss of diversity also restricts the number of unique clones that could establish colonization during transmission between hosts. Moreover, we show that genetic variation in the blp locus is associated with transmissibility in the human population. We posit this is due to its importance in clonal selection and its role as a selfish element.

Background : Before conducting an MRI examination, there will be a positioning. One of the positions carried out is the placement of isocenter. Isocenter is a magnetic center point where each gradient will be centered at that midpoint (Brown and Semelka, 2010). Different isocenter placements can affect image quality (Caramanos et al, 2010). In practical experience in the Telogorejo Semarang SMC Hospital, the placement of the isocenter in the nation while in Dr. Soetomo and Premier Surabaya Hospital placed Isocenter in Glabella. To examine the effect of isocenter placement on image quality and MRI brain T1 fast spin echo anatomical image information in the nasion and glabella and determine the optimal isocenter placement between the nasion and glabella. Method : This type of research is quantitative with an experimental approach. This research was conducted at Telogorejo SMC Hospital Semarang. Data in the form of 16 MRI images of sagittal brain pieces of T1 Fast Spin Echo weighting on isocenter placement in the nasion and glabella of 8 volunteers. Testing is done using the SPSS program where the image quality is analyzed by linear regression test, while for anatomical image information is analyzed by the Wilcoxon test. Result : The results of this study indicate that there is an effect of isocenter placement on the quality of MRI brain images of T1 fast spin echo sagittal pieces as seen from p value SNR (0.039) and CNR (0.010)

Membrane lipid homeostasis is required for bacteria to survive in a spectrum of host environments. This homeostasis is achieved by regulation of fatty acid chain length and of the ratio of unsaturated to saturated fatty acids. In the pathogen Streptococcus pneumoniae, fatty acid biosynthesis is carried out by a cluster of fatty acid biosynthesis (fab) genes (FASII locus) whose expression is controlled by the FabT repressor. Encoded immediately downstream of the FASII locus is BriC, a competence-induced, cell-cell communication peptide that promotes biofilm development as well as nasopharyngeal colonization in a murine model of pneumococcal carriage. Here, we demonstrate that briC is cotranscribed with genes of the fab gene cluster and that a reduction of briC levels, caused by decoupling its transcription from fab gene cluster, negatively affects biofilm development. BriC elevates fabT transcription, which is predicted to alter the balance of unsaturated and saturated fatty acids produced by the pathway. We find that briC inactivation results in a decreased production of unsaturated fatty acids. This affects the membrane properties by decreasing the abundance of di-unsaturated phosphatidylglycerol molecular species. We propose that the link between BriC, FabT, and phospholipid composition contributes to the ability of S. pneumoniae to alter membrane homeostasis in response to the production of a quorum-sensing peptide. IMPORTANCE Adaptation of bacteria to their host environment is a key component of colonization and pathogenesis. As an essential component of bacterial membranes, fatty acid composition contributes to host adaptation. Similarly, cell-cell communication, which enables population level responses, also contributes to host adaptation. While much is known about the pathways that control the biosynthesis of fatty acids, many questions remain regarding regulation of these pathways and consequently the factors that affect the balance between unsaturated and saturated fatty acids. We find that BriC, a cell-cell communication peptide implicated in biofilm regulation and colonization, both is influenced by a fatty acid biosynthesis pathway and affects this same pathway. This study identifies a link between cell-cell communication, fatty acid composition, and biofilms and, in doing so, suggests that these pathways are integrated into the networks that control pneumococcal colonization and host adaptation.

Significance During infection, microbes must survive the hostile environmental conditions of the human host. When exposed to stresses, bacteria activate an intracellular response, known as the stringent response pathway, to ensure their survival. This study connects two fundamental pathways important for cellular growth in a Gram-positive pathogen: it demonstrates that enzymes responsible for cell wall biosynthesis are connected to the stringent response pathway via their ability to ameliorate errors in protein translation. Our study was performed on S. pneumoniae, where the MurM cell wall biosynthesis enzyme, a tRNA-dependent amino acyl transferase, is linked to penicillin resistance. We now demonstrate the importance of MurM in translation quality control and establish that it serves as a gatekeeper of the stringent response pathway., Survival in the human host requires bacteria to respond to unfavorable conditions. In the important Gram-positive pathogen Streptococcus pneumoniae, cell wall biosynthesis proteins MurM and MurN are tRNA-dependent amino acyl transferases which lead to the production of branched muropeptides. We demonstrate that wild-type cells experience optimal growth under mildly acidic stressed conditions, but ΔmurMN strain displays growth arrest and extensive lysis. Furthermore, these stress conditions compromise the efficiency with which alanyl-tRNAAla synthetase can avoid noncognate mischarging of tRNAAla with serine, which is toxic to cells. The observed growth defects are rescued by inhibition of the stringent response pathway or by overexpression of the editing domain of alanyl-tRNAAla synthetase that enables detoxification of tRNA misacylation. Furthermore, MurM can incorporate seryl groups from mischarged Seryl-tRNAAlaUGC into cell wall precursors with exquisite specificity. We conclude that MurM contributes to the fidelity of translation control and modulates the stress response by decreasing the pool of mischarged tRNAs. Finally, we show that enhanced lysis of ΔmurMN pneumococci is caused by LytA, and the murMN operon influences macrophage phagocytosis in a LytA-dependent manner. Thus, MurMN attenuates stress responses with consequences for host–pathogen interactions. Our data suggest a causal link between misaminoacylated tRNA accumulation and activation of the stringent response. In order to prevent potential corruption of translation, consumption of seryl-tRNAAla by MurM may represent a first line of defense. When this mechanism is overwhelmed or absent (ΔmurMN), the stringent response shuts down translation to avoid toxic generation of mistranslated/misfolded proteins.

Membrane lipid homeostasis is required for bacteria to survive in a spectrum of host environments. This homeostasis is achieved by regulation of fatty acid chain length and of the ratio of saturated to unsaturated fatty acids. In the pathogen Streptococcus pneumoniae, fatty acid biosynthesis is encoded by a cluster of fatty acid biosynthesis (fab) genes (FASII locus) whose expression is controlled by the FabT repressor. Encoded immediately downstream of the FASII locus is BriC, a competence-induced, cell-cell communication peptide that promotes biofilm development as well as nasopharyngeal colonization in a murine model of pneumococcal carriage. Here, we demonstrate that briC is co-transcribed with genes of the fab gene cluster and that a reduction of briC levels, caused by decoupling its transcription from fab gene cluster, negatively impacts biofilm development. BriC elevates fabT transcription, which is predicted to alter the balance of saturated and unsaturated fatty acids produced by the pathway. We find that briC inactivation results in a decreased production of unsaturated fatty acids that impact the membrane properties by decreasing the abundance of di-unsaturated phosphatidylglycerol molecular species. We propose that the link between BriC, FabT and phospholipid composition contributes to the ability of S. pneumoniae to alter membrane homeostasis in response to the production of a quorum-sensing peptide.IMPORTANCEAdaptation of bacteria to their host environment is a key component of colonization and pathogenesis. As an essential component of bacterial membranes, fatty acid composition contributes to host adaptation. Similarly, so does cell-cell communication, which serves as a mechanism for population levels responses. While much is known about the pathways that control the biosynthesis of fatty acids, many questions remain regarding regulation of these pathways and consequently the factors that impacts the balance between saturated and unsaturated fatty acids. We find that BriC, a cell-cell communication peptide implicated in biofilm regulation and colonization, is both influenced by a fatty acid biosynthesis pathway and impacts this same pathway. This study identified a link between cell-cell communication, fatty acid composition, and biofilms and, in doing so, suggests that these pathways are integrated into the networks that control pneumococcal colonization and host adaptation.

Physical based creep models that elucidate the creep deformation behaviour with ongoing microstructural evolution can be a useful tool for the components life assessment as well as the design of improved materials, deployed at high temperature and pressure. In this research work, a creep model that is a combination of physical based model and CDM approach is employed to predict the creep curves of steel 316LN. The microstructure based variables those are different dislocation densities (mobile and forest) are the input parameters. The model provides a provision for the assessment of each microstructural variable, in each time step of the creep deformation. Consequently, other than creep curves, the model also demonstrates the evolution of dislocation density (mobile and forest), dislocation velocity, dislocation mobility and mean free path. Initial values of input parameters (various dislocation densities and mean free path) are obtained from the literature for initializing the model. It can be observed that the predicted creep curves are in reasonable agreement with experimental ones and the evolution of other involved parameters is discussed thoroughly.

International scientists, engineers, and students have made and continue to make significant contributions to the U.S.’s science and engineering research enterprise. This source of talent is a comparative advantage for the U.S. and is critical for keeping the nation at the leading edge of discovery and knowledge. This paper quantifies and reaffirms the impact of international scientists and scholars, who serve as a vital source of talent fueling American ingenuity, innovation, and leadership. Bolstering measures to attract and retain top students from across the globe and fostering a culture where immigrants are welcome and can thrive is imperative for the continued success of the U.S.’s society and economy.

The stunning ability of bacteria to evolve and adapt has contributed to the success of these single cells, which have inhabited the Earth for billions of years and play vital roles in the environment and in human health. The goal of this chapter is to present and discuss the population-level organizational scheme of bacterial pangenomes, wherein genes are distributed among the strains of a species, such that each individual strain encodes only a subset of the genes available at the population level. Genes from the accessory/distributed genome (those present only in a subset of strains within a species) impart diverse functions or variations on a conserved function to strains. Moreover, horizontal gene transfer generates novel gene combinations. The maintenance and spread of any given gene arrangement are influenced by fitness. Further, the extent of genomic plasticity is regulated by restriction modification systems, phage-defense systems, and Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)—associated proteins (CRISPR-Cas). The combination of a pangenome structure and genomic plasticity reveals a successful strategy for bacterial adaptation to ever-changing environments. From a clinical perspective, pangenome analyses inform the selection of therapeutic targets, designed to focus either on an entire species or on virulence features within a species. Further, they provide a framework for modeling the efficacy of drugs and vaccines. In summary, following the explosion in sequencing technology, pangenome studies have revealed remarkable genomic organizations at the levels of species, with important implications to our understanding of evolution, and our ability to design therapeutics and predict their long-term outcomes.

Bacterial cells modify their gene expression profiles throughout different stages of growth and in response to environmental cues. Analyses of gene expression across conditions reveal both conserved and condition-specific gene responses of bacteria to adapt to these dynamic conditions. In this chapter, we present a guide to pneumococcal RNA extraction for use in the NanoString nCounter platform. The nCounter is a highly effective method to measure gene expression of bacteria not only in a planktonic mode of growth but also in the presence of host cells where the RNA of interest represents only a small portion of the total material.

Binding of Streptococcus pneumoniae (Spn) to nasal mucus leads to entrapment and clearance via mucociliary activity during colonization. To identify Spn factors allowing for evasion of mucus binding, we used a solid-phase adherence assay with immobilized mucus of human and murine origin. Spn bound large mucus particles through interactions with carbohydrate moieties. Mutants lacking neuraminidase A (nanA) or neuraminidase B (nanB) showed increased mucus binding that correlated with diminished removal of terminal sialic acid residues on bound mucus. The non-additive activity of the two enzymes raised the question why Spn expresses two neuraminidases and suggested they function in the same pathway. Transcriptional analysis demonstrated expression of nanA depends on the enzymatic function of NanB. As transcription of nanA is increased in the presence of sialic acid, our findings suggest that sialic acid liberated from host glycoconjugates by the secreted enzyme NanB induces the expression of the cell-associated enzyme NanA. The absence of detectable mucus desialylation in the nanA mutant, in which NanB is still expressed, suggests that NanA is responsible for the bulk of the modification of host glycoconjugates. Thus, our studies describe a functional role for NanB in sialic acid sensing in the host. The contribution of the neuraminidases in vivo was then assessed in a murine model of colonization. Although mucus-binding mutants showed an early advantage, this was only observed in a competitive infection, suggesting a complex role of neuraminidases. Histologic examination of the upper respiratory tract demonstrated that Spn stimulates mucus production in a neuraminidase-dependent manner. Thus, an increase production of mucus containing secretions appears to be balanced, in vivo, by decreased mucus binding. We postulate that through the combined activity of its neuraminidases, Spn evades mucus binding and mucociliary clearance, which is needed to counter neuraminidase-mediated stimulation of mucus secretions., Author summary Streptococcus pneumoniae (Spn) is a leading mucosal pathogen, whose host interaction begins with colonization of the upper respiratory tract. While there has been extensive investigation into bacterial interaction with epithelial cells, there is little understanding of bacterial-mucus interactions. Our study used mucus of human and murine origin and a murine model of colonization to study mucus associations involving Spn. The main findings reveal i) the enzymatic activity of Spn’s neuraminidases (NanA and NanB) contribute to mucus evasion through removing terminal sialic acid, ii) the enzymatic activity of NanB controls expression of the main neuraminidase, NanA, and iii) Spn induces sialic acid containing mucus secretions in vivo in a neuraminidase-dependent manner. We postulate that during colonization, neuraminidase-dependent reduction in mucus binding enables evasion of mucociliary clearance, which is necessary to counter neuraminidase-mediated stimulation of mucus secretions. Thus, our study provides new insights into the role of Spn neuraminidases on colonization.

Gram-positive bacteria employ an array of secreted peptides to control population-level behaviors in response to environmental cues. We review mechanistic and functional features of secreted peptides produced by the human pathogen Streptococcus pneumoniae. We discuss sequence features, mechanisms of transport, and receptors for 3 major categories of small peptides: the double-glycine peptides, the Rap, Rgg, NprR, PlcR, and PrgX (RRNPP)-binding peptides, and the lanthionine-containing peptides. We highlight the impact of factors that contribute to carriage and pathogenesis, specifically genetic diversity, microbial competition, biofilm development, and environmental adaptation. A recent expansion in pneumococcal peptide studies reveals a complex network of interacting signaling systems where multiple peptides are integrated into the same signaling pathway, allowing multiple points of entry into the pathway and extending information content in new directions. In addition, since peptides are present in the extracellular milieu, there are opportunities for crosstalk, quorum sensing (QS), as well as intra- and interstrain and species interactions. Knowledge on the manner that population-level behaviors contribute to disease provides an avenue for the design and development of anti-infective strategies.

Bacterial cells modify their gene expression profiles throughout different stages of growth and in response to environmental cues. Analyses of gene expression across conditions reveal both conserved and condition-specific gene responses of bacteria to adapt to these dynamic conditions. In this chapter, we present a guide to pneumococcal RNA extraction for use in the NanoString nCounter platform. The nCounter is a highly effective method to measure gene expression of bacteria not only in a planktonic mode of growth but also in the presence of host cells where the RNA of interest represents only a small portion of the total material.

Botrytis bunch rot, caused by Botrytis cinerea, is one of the most destructive diseases of grapes (Vitis vinifera L.). A phenotyping assay was designed under laboratory conditions to analyze grape-B. cinerea interactions. A conidial suspension of concentration 1 × 105 conidia/ml was prepared and used to inoculate berries from two different grape varieties: V. aestivalis-derived ‘Norton’, a North American hybrid, and V. vinifera ‘Cabernet Sauvignon’, a European grape. The differences in the infection reaction between these two cultivars were visible 10 days after inoculation. Berries from Norton had a low average disease incidence (7.5 %) and disease severity (3.7 %). In contrast, berries from Cabernet Sauvignon were completely susceptible, exhibiting an average disease incidence and severity greater than 90 %. This phenotyping assay can be further applied to evaluate disease progression in a mapping population, developed by crossing V. aestivalis-derived ‘Norton’ and V. vinifera ‘Cabernet Sauvignon’.

Evaporative silicification can drive microbial fossilization within diverse natural habitats. Research into this process is pivotal to understanding the terrestrial fossil record and the preservation of biomarkers within extreme environments. We employed laboratory experiments to silicify the polyextremotolerant bacterium Halomonas hydrothermalis cultured at low, intermediate and high salinities (1, 3.5 or 11.8% w/v of NaCl) under iron-rich or iron-deprived conditions. Silicification was achieved by adding sodium silicate solution (30 or 150 ppm of Si) onto cultures, followed by evaporation. Scanning electron microscopy demonstrated the presence of mineralized bacteria with intact morphology across all culture conditions. However, multivariate analysis of the attenuated total reflectance Fourier-transform infrared (ATR-FT-IR) spectra of silicified cultures showed significant differences between the examined salinities, most notably between cultures silicified after incubation at high salinity and ...

Grape bunch rot, caused by the necrotrophic fungal pathogen Botrytis cinerea, is one of the most destructive diseases in grapevines worldwide. At this point, control of Botrytis bunch rot is achieved through a combination of canopy management and fungicide usage. The identification of resistant germplasm could result in improved disease management. The Vitis aestivalis-derived ?Norton? is one of the very few commercial red grape cultivars that can be grown under elevated B. cinerea pressure where highly susceptible V. vinifera is difficult to grow. Tests on the F1 progeny from a cross between V. aestivalis-derived ?Norton? and V. vinifera ?Cabernet Sauvignon? may elucidate the underlying genetic and molecular mechanisms of berry disease resistance and berry quality. To this end, we have developed a population of 92 F1 genotypes from this cross that has yielded fruit for the past three years. Also, a linkage map employing simple sequence repeat (SSR) markers clustered in 19 chromosomes has been under construction. By using genotyping-by-sequencing (GBS) technology as part of the VitisGen (www.vitisgen. org) project, we aim to improve marker-density and identify the tightly linked markers associated with Botrytis bunch rot resistance. The overall goals of this study are to use genetic markers to rapidly implement favorable alleles and accelerate breeding cycles for new improved cultivar releases that warrant decreased pesticide use in the vineyard, resulting in greater environmental and economic sustainability.

The Vitis aestivalis -derived ‘Norton’ is one of the very few commercial red grape varieties that can be grown in regions with high disease pressure and cold temperatures in winter and spring where V. vinifera is difficult to grow. This study began with the objectives of generating interspecific hybrids of Norton and V. vinifera (Cabernet Sauvignon, Pinot Noir, Syrah, and Merlot) in order to combine the disease resistance and cold hardiness characters of V. aestivalis with the excellent wine quality of V. vinifera . Norton grapes were also crossed with the Vitis interspecific hybrid ‘Vignoles’ for the same reason. The true hybrids obtained from all of the crosses were identified using microsatellite markers. A high percentage of hybrids (70–92%) were obtained in all crosses except Norton × Merlot (17%). Results demonstrate the reliability of SSR markers for Norton-based interspecific hybrid identification.

Fluidized beds have widespread industrial applications ranging from chemical industries to power plants. The flow inside a fluidized bed system consists of two main phases, a particle phase and the fluid phase. The two phases are strongly coupled to each other through various forces like drag and pressure. Capturing this multiphase phenomenon requires modeling strategies that possess good fidelity over a range of scales. Discrete Element Modeling (DEM) coupled with Computational Fluid Dynamics (CFD) provides a good platform to analyze the complex coupled multiphase hydrodynamics inside fluidized bed systems. Conventional DEM-CFD framework suffers from contradictory spatial resolution requirements for the particle and fluid phases, respectively. This prevents the conventional DEM-CFD method to be applied to geometries that have features comparable to the particle diameter of the solid phase. The novelty of this work lies in the development and validation of a two-grid formulation that removes the resolution restrictions of the conventional DEM-CFD framework. The results obtained from this new framework agree reasonably well with the experiments showing the capability of the new scheme to simulate conditions not possible with conventional DEM-CFD framework. In addition, this research also focuses on performing both 2D and 3D jetting fluidized bed simulations having millions of particles; validate/compare results with experiments and to perform heat transfer studies in a jetting fluidized bed system. The results suggest convective and diffusive mixing for a single jet at higher superficial velocity to be better than the mixing obtained in a multiple jet framework. The comparison with experimental results obtained in a multiple jetting setup shows that a 2D simulation captures the essential jet characteristics near the distributor plate reasonably well while a 3D simulation is needed to capture proper bubble dynamics near the freeboard of the bed. These results give insight into the detailed dynamics of fluidized bed systems and provide a foundation for a better design of these systems. Ph. D.