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For plants to develop, fertile soil is necessary. Estimating soil parameters based on time change is crucial for enhancing soil fertility. Sentinel-2's remote sensing technology produces images that can be used to gauge soil parameters. In this study, values for soil parameters such as electrical conductivity, pH, organic carbon, and nitrogen are derived using Sentinel-2 data. In order to increase the clustering accuracy, this study suggests using Canopy centre-based fuzzy-C-means clustering and comparing it to manual labelling and other clustering techniques such as Canopy, density-based, expectation-maximisation, farthest-first, k-means, and fuzzy-C-means clustering, its usefulness is demonstrated. The proposed clustering achieved the highest clustering accuracy of 78.42%. Machine learning-based classifiers were applied to classify soil fertility, including Naive Bayes, support vector machine, decision trees, and random forest (RF). Dataset labelled with the proposed RF clustering classifier achieves a high classification accuracy of 99.69% with ten-fold cross-validation.

AbstractObjectiveHigh level of exogenous ROS in the circulation affects RBC membrane integrity which facilitates the generation of endogenous RBC ROS, implicated in series of physiological changes primarily associated with thrombosis and vital tissue damage. Although, Pennisetum glaucum(pearl millet)stores abundance of proteins, their therapeutic potential is least explored. Thus, the purpose of this study is to examine the role of Pennisetum GlaucumProtein Extract (PGE) on oxidative stress induced cell/tissue damage and thrombosis.MethodIn this investigation, protein characterization was done by using SDS-PAGE, Native-PAGE, PAS-staining and HPLC. In-vitrooxidative stress was induced in RBC using sodium nitrite. While, in-vivooxidative stress was induced in experimental rats using diclofenac. Stress markers and biochemical parameters were evaluated. Role of PGE on thrombosis was assessed by using, in-vitroplasma recalcification time, activated partial thromboplastin time, prothrombin time, mouse tail bleeding time (In-vivo) and platelet aggregation.Results:PGE revealed varied range of molecular weight proteins on SDS-PAGE. PGE normalized the sodium nitrite induced oxidative damage of RBC and diclofenac induced oxidative damage in liver, kidney and small intestine. PGE exhibited anticoagulant effect by increasing the coagulation time of both PRP and PPP and mouse tail bleeding time. Furthermore, PGE prolonged the clotting time of only APTT but did not affect PT. PGE inhibited agonists ADP and epinephrine induced platelet aggregation.ConclusionOur findings suggest, PGE could be a better contender in the management of oxidative stress and its associated diseases.AbbreviationsPGEPennisetum Glaucum protein ExtractAPPTActivated Partial Thromboplastin TimePTProthrombin TimeROSReactive Oxygen SpeciesPRPPlatelet Rich PlasmaPPPPlatelet Poor PlasmaSDS-PAGESodium Dodecyl Sulfate-Polyacrylamide Gel ElectrophoresisPASPeriodic Acid-schiff StainingODOptical DensityINRInternational Normalized RatioPBSPhosphate Buffered SalineSODSuperoxide DismutaseTCATrichloro Acetatic AcidDTNBDi-Thio-bis-NitroBenzoic acidSGOTSerum Glutamate Oxaloacetate TransaminaseSGPTSerum Glutamate Pyruvate TransaminaseALPAlkaline PhosphataseDFCDiclofenacSylSilymarinMEDMinimum Edema DoseMHDMinimum Hemorrhagic Dose

Biomolecular condensates formed by liquid–liquid phase separation have been implicated in multiple diseases. Modulation of condensate dynamics by small molecules has therapeutic potential, but so far, few condensate modulators have been disclosed. The SARS-CoV-2 nucleocapsid (N) protein forms phase-separated condensates that are hypothesized to play critical roles in viral replication, transcription, and packaging, suggesting that N condensation modulators might have anti-coronavirus activity across multiple strains and species. Here, we show that N proteins from all seven human coronaviruses (HCoVs) vary in their tendency to undergo phase separation when expressed in human lung epithelial cells. We developed a cell-based high-content screening platform and identified small molecules that both promote and inhibit condensation of SARS-CoV-2 N. Interestingly, these host-targeted small molecules exhibited condensate-modulatory effects across all HCoV Ns. Some have also been reported to exhibit antiviral activity against SARS-CoV-2, HCoV-OC43, and HCoV-229E viral infections in cell culture. Our work reveals that the assembly dynamics of N condensates can be regulated by small molecules with therapeutic potential. Our approach allows for screening based on viral genome sequences alone and might enable rapid paths to drug discovery with value for confronting future pandemics.

The aim of the present study is to synthesize iron oxide nanoparticles (FeONPs) and to evaluate the antibacterial efficacy against selected bacteria from aqueous leaf extract of Chamaecrista nigricans, a medicinal under shrub. The precursor for the synthesis of FeONPs was 0.01 M Ferric chloride hexahydrate (FeCl3.6H2O). The structural and optical characterization of FeONPs were performed using ultra violet-visible spectroscopy (UV–vis), Fourier transform infrared (FTIR) spectroscopy, X-ray diffraction (XRD), scanning electron microscopy (SEM), and field emission scanning electron microscopy with energy dispersive X-ray spectroscopy (FESEM-EDX). The effect of antibacterial activity was envisaged with the synthesized FeONPs by means of the agar well diffusion method against three gram-negative bacterial pathogens, namely, Escherichia coli(MTCC 1610), Klebsiella pneumoniae(MTCC618), and Pseudomonas aeruginosa(MTCC 741). The effectiveness of FeONPs was determined by measuring the zones of inhibition in millimeter (mm). UV–visible spectroscopy at 238 nm clearly indicates the presence of FeONPs. The FTIR spectroscopic analysis verified the existence of metallic ions of Fe and bioactive components derived from the plant. The X-ray diffraction technique was employed to ascertain the phase and crystal structure. The results obtained from FESEM-EDX demonstrated that the nanoparticles of iron oxide exhibited a spherical morphology, with a mean diameter of 17 nm in terms of surface volume. Antibacterial investigation has revealed that the FeONPs has greater efficacy in inhibiting the growth of Escherichia coliwhen compared to Pseudomonas aeruginosa.

Problem statement: Antibiotic susceptibility is still the best way for bacterial pathogen escape mechanism against immunity. Approach: In the present investigation, bacterial pathogens like Staphylococcus aureus, Escherichia coli, Aeromonas hydrophila, Klebsiella and Pseudomonas aeruginosa were used to screen antibiotic susceptibility and immunomodulatory potential. Results: All the test pathogens were sensitive to all the test antibiotics 11 [+ or -] 2 mm) except penicillin. The conditions for the preparation of antigens of intact natural composition and conformation from pathogens (whole cell and heat killed), were determined using Swiss albino mice (Balb/C) as experimental species. Immunomodulatory potential of test pathogens were screened using animal model. Test pathogen decreases the body weight comparing that of normal mice, some notable changes were also noted in activity, growth, water consumption, feed consumption. Antibody titre level in animal serum decreased upto 50% in whole cell pathogen and heat killed pathogen treated animals. Conclusion: The five pathogens administered animals, decrement in B-lymphocyte was much pronounced in Pseudomonas aeruginosa followed by Escherichia coli, Staphylococcus aureus, Klebsiella sp., Aeromonas hydrophila in the 5 week. Pathogen treated mice showed an IgG suppressive effect. It is found to be suppressive to T cell production, so induction in cell mediated immunity has confirmed pathogenic potential of test pathogens. All these test pathogenic strains were remarkably suppressing immune system of pathogen exposed animals. Key words: Bacterial pathogens, immune response, antibiotic susceptibility, immunomodulatory, mice, INTRODUCTION Pathogens expresses many potential virulence factors such as, surface proteins that promote colonization of host tissues; invasins that promote bacterial spread in tissues (leukocidin, kinases, hyaluronidase); surface factors that [...]

Hematopoietic progenitor kinase 1 (HPK1) is an MAP4K family member within the Ste20-like serine/threonine branch of the kinome. HPK1 expression is limited to hematopoietic cells and has a predominant role as a negative regulator of T cell function. Because of the central/dominant role in negatively regulating T cell function, HPK1 has long been in the center of interest as a potential pharmacological target for immune therapy. The development of a small molecule HPK1 inhibitor remains challenging because of the need for high specificity relative to other kinases, including additional MAP4K family members, that are required for efficient immune cell activation. Here, we report the identification of the selective and potent HPK1 chemical probe, A-745. In unbiased cellular kinase-binding assays, A-745 demonstrates an excellent cellular selectivity binding profile within pharmacologically relevant concentrations. This HPK1 selectivity translates to an in vitroimmune cell activation phenotype reminiscent of Hpk1-deficient and Hpk1-kinase-dead T cells, including augmented proliferation and cytokine production. The results from this work give a path forward for further developmental efforts to generate additional selective and potent small molecule HPK1 inhibitors with the pharmacological properties for immunotherapy.

In today's phase of growing technology Convolution Neural Networks (CNNs) are all over the places. It is a thriving segment in machine learning and Artificial Intelligences (AI) techniques. CNN needs bulk amount of computing competence and memory with higher frequency range. In this present investigation, Pre-Accumulator and Post-Multipliers (PAPM) are proposed which accelerate the speed of processor. 4-bit multiplier using Carry Save Adder (CSA) is built with 6Transistors-Adder and sutras of Vedic mathematics is constructed. Accumulator of multiplier and accumulator are designed with Two Level Edge Triggering Flip-Flops (TLET-FF) to increase bandwidth of the memory. The proposed architecture of Multiply Accumulate (MAC) circuit consumes very less power when compared to existing high speed MACs. Performance of accumulator is contrasted with three different, two-level triggered flip-flops namely 16TLET-FF, 14TLET-FF and 12TLET-FFs. The projected MAC replaces the existing multipliers due its low power together with high frequency of operation.

Wideband slot antenna gain enhancement using concentric square and circular loop metasurface reflector (SCLMR) is presented in this paper. In two steps, the design is realized. The analysis design of a FSS-based SCLMR unit cell is included in the first step. The SCLMR structure is made up of resonant metallic square-loop and inner circular loop structures. In terms of bandwidth, the unit cell structure can be considered as a stop-band filter (2.5 GHz-6.5 GHz). The design of a wideband slot antenna (29 mm ×24 mm ×1.57 mm) is included in the second step. EM simulation tool is used to analyze, design, optimize, and validate the proposed unit cell and slot antenna combination. CST microwave studio is utilized to design and analyze a slot antenna and a unit cell in this study. The computed result of an antenna with SCLMR indicates a 101.21 % operational impedance bandwidth (2.76 GHz to 8.41 GHz). With SCLMR, the antenna has a high gain of 8.75 dBi. The designed antenna structure provides a gain enhancement of 5.50 dBi while in comparison to the same antenna without SCLMR.

Near infrared (NIR) spectroscopy is developing as an advanced and non-invasive tool in the wood, wood products and forestry sectors. It may be applied as a rapid and cost effective technique for assessment of different wood quality parameters of timber species. In the present study, NIR spectra of heartwood samples of Tectona grandis (teak) were collected before measuring fibre morphological parameters (fibre length, fibre diameter and fibre lumen diameter)and main chemical constituents (cellulose, hemicellulose, lignin and extractives) using maceration and wet chemistry methods respectively. Multivariate partial least squares (PLS) regression was applied to develop the calibration models between measured values of wood parameters and NIR spectral data. Pre-processing of NIR spectra demonstrated better predictions based on higher values of correlation coefficient for estimation (R^2), validation (R_cv^2), ratio of performance to deviation (RPD), and lower values of root mean square errors of estimation (RMSEE), cross-validation (RMSECV) and number of latent variable (rank). Internal cross-validation was used to find the optimum rank. Robust calibrations models with high R^2 (>0.87), low errors and high RPD values (> 2.93) were observed from PLS analysis for fibre morphological parameters and main chemical constituents of teak. These linear models may be applied for rapid and cost effective estimation of different fibre parameters and chemical constituents in routine testing and evaluation procedures for teak.

Mass spectrometry (MS) can provide high sensitivity and specificity for biochemical assays without the requirement of labels, eliminating the risk of assay interference. However, its use had been limited to lower-throughput assays due to the need for chromatography to overcome ion suppression from the sample matrix. Direct analysis without chromatography has the potential for high throughput if sensitivity is sufficient despite the presence of a matrix. Here, we report and demonstrate a novel direct analysis high-throughput MS system based on infrared matrix-assisted desorption electrospray ionization (IR-MALDESI) that has a potential acquisition rate of 33 spectra/s. We show the development of biochemical assays in standard buffers for wild-type isocitrate dehydrogenase 1 (IDH1), diacylglycerol kinase zeta (DGKζ), and p300 histone acetyltransferase (P300) to demonstrate the suitability of this system for a broad range of high-throughput lead discovery assays. A proof-of-concept pilot screen of ∼3k compounds is also shown for IDH1 and compared to a previously reported fluorescence-based assay. We were able to obtain reliable data at a speed amenable for high-throughput screening of large-scale compound libraries.

Alzheimer’s Disease (AD) is a progressive neurodegenerative disease and the most common cause of dementia. The current treatment options for AD are limited to ameliorating cognitive decline temporarily and not reversing or preventing the progression of dementia. Hence, more effective therapeutic strategies are needed to combat this devastating disease. The low-density lipoprotein receptor has been shown to modulate the neuronal metabolism of cholesterol and apolipoprotein E, a major genetic risk factor for AD. LDLR overexpression in mice has been shown to increase amyloid-β clearance and reduce amyloid deposition. We conducted a phenotypic screen to identify novel signaling pathways and targets that regulate LDLR expression in glial cells using an annotated compound library of approximately 29 000 compounds. The screen identified novel targets such as polo like kinase 1 (PLK1), activin receptor like kinase 5 (ALK5), and serotonin transporter (SERT). We used genetic, chemical biology and pathway analysis to confirm the target hypothesis. This work highlights that phenotypic screening is a promising strategy to identify novel mechanisms and targets for therapeutic intervention of complex neurodegenerative disorders.

Background: Congenital central nervous system (CNS) anomalies are the structural or functional abnormalities of the brain and spinal cord that occur during the intrauterine developmental process.Purpose: The present study aims to detect the prevalence of congenital CNS anomalies among stillborn fetuses, the association between congenital anomalies and maternal factors, and also the association between autopsy and ultrasound findings.Methods: This study was conducted on 50 stillborn fetuses, obtained from the Department of Obstetrics and Gynecology at JSS Medical College and Hospital, Mysuru. The fetuses were fixed in 10% formalin and autopsies were performed as per the standard fetal autopsy protocol. The congenital CNS anomalies were studied in detail.Results: CNS anomalies were the most common congenital anomalies observed. Out of the total 50 stillborn fetuses studied, CNS anomalies were found in 17 fetuses and their occurrence was more common among male stillborn than females. Meningomyelocele was the most common anomaly, followed by anencephaly. The other anomalies documented were meningocele, encephalocele, meningoencephalocele, agenesis of the corpus callosum, craniorachischisis, bifid cerebellum with hypoplastic vermis, holoprosencephaly, and sirenomelia. Fisher’s exact test showed a significant association between maternal hypothyroidism and congenital CNS anomalies (P< .05). The autopsy confirmed the ultrasound findings in 40 (80%) fetuses. There were significant additional findings observed in seven (14%) fetal autopsies and ultrasound diagnosis completely changed in three (6%) cases, after the final autopsy procedure.Conclusion: The fetal autopsy is the single most directly evident investigation, which gives information that changes or significantly improves the clinical diagnosis. A multidisciplinary holistic approach toward pregnancy will help to detect any kind of abnormality in the fetus and thus to take a timely decision toward the management.

The SARS-CoV-2 main protease (Mpro) is essential for viral replication because it is responsible for the processing of most of the non-structural proteins encoded by the virus. Inhibition of Mpro prevents viral replication and therefore constitutes an attractive antiviral strategy. We set out to develop a high-throughput Mpro enzymatic activity assay using fluorescently labeled peptide substrates. A library of fluorogenic substrates of various lengths, sequences and dye/quencher positions was prepared and tested against full length SARS-CoV-2 Mpro enzyme for optimal activity. The addition of buffers containing strongly hydrated kosmotropic anion salts, such as citrate, from the Hofmeister series significantly boosted the enzyme activity and enhanced the assay detection limit, enabling the ranking of sub-nanomolar inhibitors without relying on the low-throughput Morrison equation method. By comparing cooperativity in citrate or non-citrate buffer while titrating the Mpro enzyme concentration, we found full positive cooperativity of Mpro with citrate buffer at less than one nanomolar (nM), but at a much higher enzyme concentration (∼320 nM) with non-citrate buffer. In addition, using a tight binding Mpro inhibitor, we confirmed there was only one active catalytical site in each Mpro monomer. Since cooperativity requires at least two binding sites, we hypothesized that citrate facilitates dimerization of Mpro at sub-nanomolar concentration as one of the mechanisms enhances Mpro catalytic efficiency. This assay has been used in high-throughput screening and structure activity relationship (SAR) studies to support medicinal chemistry efforts. IC50values determined in this assay correlates well with EC50values generated by a SARS-CoV-2 antiviral assay after adjusted for cell penetration.

Over the last 5 years, IR-MALDESI-MS (Infrared Matrix-Assisted Laser Desorption Electrospray Ionization Mass Spectrometry) has been demonstrated for use in a range of high-throughput biochemical and cellular assays with remarkable sample acquisition rates up to 22 Hz for a single 384-well assay plate. With such high single plate acquisition rates, the rate limiting step becomes how fast subsequent plates can be presented to the MS for analysis. To make this transfer as fast as possible while maintaining safe operation in a laboratory environment, we developed a collaborative robotic plate transfer system (CRPTS) that combines a 6-axis robot with dual plate grippers, a 7thaxis conveyor stage, and a 420-plate capacity sample loading window. As a demonstration of the throughput and flexibility of CRPTS, we performed a biochemical assay that monitored the oxidation of tris(2-carboxyethyl)phosphine (TCEP) to screen for nuisance compounds. Using continuous and step motion scan profiles, we analyzed 158,799 compounds contained in 448 assay plates over the course of 12.5 hours (Z-Factor=0.87) and 17.5 hours (Z-factor=0.99), respectively. Extrapolating these results enables the screening of a million compounds within 6-7 working days.

The continuous and indiscriminate use of NPK fertilizers for boosting productivity in the farming sector longer periods resulted in the imbalance of soil nutrients in the long run. This nutrient disparity in soil gradually throw back in crops, animals and human beings, leading to various degenerative and deficiency related diseases now more than ever. This constrained site specific nutrient management for crops, which essentially rely upon evaluation of variability spatially and temporarily. In this study we scrutinized the spatial variation of soil fertility in exhaustive plough lands of Thrissur district, Kerala, India. A total of 600 geo referred soil samples were collected from different agroecological units of the district and examined for selected micronutrients such as Fe, Cu, Mn, Zn and B. Geo-statistical mapping tool (Arc GIS10.2.2) was used to quantify the degree of spatial variability in various soil fertility parameters. The spatial variation of nutrients in the study area was assessed by using semivariogram method in kriging interpolation and spatial dependence was calculated. The best fit model was applied to the kriging interpolation according to the determination coefficient, which is the correlation of measured and predicted values on space and spatial distribution maps of all the micronutrients. Among the variables analyzed, B revealed strong spatial dependence (24%), Zn with weak spatial dependence (78%) with model gaussian and others with moderate spatial dependence. The results from the present study call to develop a strategy for site-specific management for the parameters showing moderate spatial dependence and weak spatial dependence. But for B, showing strong spatial dependence, only uniform management is needed because it was greatly affected by the structural factors such as climate, topography and parent material. Spatial variability of soil properties is essential for precision agriculture because soil parameters with little or no spatial dependence will not be conducive to site-specific management, and will be managed on the average level only.

A HTS campaign identified compound 1, an excellent hit-like molecule to initiate medicinal chemistry efforts to optimize a dual ROCK1 and ROCK2 inhibitor. Substitution (2-Cl, 2-NH2, 2-F, 3-F) of the pyridine hinge binding motif or replacement with pyrimidine afforded compounds with a clean CYP inhibition profile. Cocrystal structures of an early lead compound were obtained in PKA, ROCK1, and ROCK2. This provided critical structural information for medicinal chemistry to drive compound design. The structural data indicated the preferred configuration at the central benzylic carbon would be (R), and application of this information to compound design resulted in compound 16. This compound was shown to be a potent and selective dual ROCK inhibitor in both enzyme and cell assays and efficacious in the retinal nerve fiber layer model after oral dosing. This tool compound has been made available through the AbbVie Compound Toolbox. Finally, the cocrystal structures also identified that aspartic acid residues 176 and 218 in ROCK2, which are glutamic acids in PKA, could be targeted as residues to drive both potency and kinome selectivity. Introduction of a piperidin-3-ylmethanamine group to the compound series resulted in compound 58, a potent and selective dual ROCK inhibitor with excellent predicted drug-like properties.

IDH1 plays a critical role in a number of metabolic processes and serves as a key source of cytosolic NADPH under conditions of cellular stress. However, few inhibitors of wild-type IDH1 have been reported. Here we present the discovery and biochemical characterization of two novel inhibitors of wild-type IDH1. In addition, we present the first ligand-bound crystallographic characterization of these novel small molecule IDH1 binding pockets. Importantly, the NADPH competitive α,β-unsaturated enone 1makes a unique covalent linkage through active site H315. As few small molecules have been shown to covalently react with histidine residues, these data support the potential utility of an underutilized strategy for reversible covalent small molecule design.

The more than 1.5 billion people who live in South Asia are correctly viewed not as a single large population but as many small endogamous groups. We assembled genome-wide data from over 2,800 individuals from over 260 distinct South Asian groups. We identified 81 unique groups, 14 of which had estimated census sizes of more than 1 million, that descend from founder events more extreme than those in Ashkenazi Jews and Finns, both of which have high rates of recessive disease due to founder events. We identified multiple examples of recessive diseases in South Asia that are the result of such founder events. This study highlights an underappreciated opportunity for decreasing disease burden among South Asians through discovery of and testing for recessive disease-associated genes.

Cellular pharmacodynamic assays are crucial aspects of lead optimization programs in drug discovery. These assays are sometimes difficult to develop, oftentimes distal from the target and frequently low throughput, which necessitates their incorporation in the drug discovery funnel later than desired. The earlier direct pharmacodynamic modulation of a target can be established, the fewer resources are wasted on compounds that are acting via an off-target mechanism. Mass spectrometry is a versatile tool that is often used for direct, proximal cellular pharmacodynamic assay analysis, but liquid chromatography-mass spectrometry methods are low throughput and are unable to fully support structure–activity relationship efforts in early medicinal chemistry programs. Infrared matrix-assisted laser desorption electrospray ionization (IR-MALDESI) is an ambient ionization method amenable to high-throughput cellular assays, capable of diverse analyte detection, ambient and rapid laser sampling processes, and low cross-contamination. Here, we demonstrate the capability of IR-MALDESI for the detection of diverse analytes directly from cells and report the development of a high-throughput, label-free, proximal cellular pharmacodynamic assay using IR-MALDESI for the discovery of glutaminase inhibitors and a biochemical assay for hit confirmation. We demonstrate the throughput with a ∼100,000-compound cellular screen. Hits from the screening were confirmed by retesting in dose–response with mass spectrometry-based cellular and biochemical assays. A similar workflow can be applied to other targets with minimal modifications, which will speed up the discovery of cell active lead series and minimize wasted chemistry resources on off-target mechanisms.

Compounds that inhibit glutathione peroxidase 4 (GPX4) hold promise as cancer therapeutics in their ability to induce a form of nonapoptotic cell death called ferroptosis. Our research identified 24, a structural analog of the potent GPX4 inhibitor RSL3, that has much better plasma stability (t1/2> 5 h in mouse plasma). The bioavailability of 24provided efficacious plasma drug concentrations with IP dosing, thus enabling in vivostudies to assess tolerability and efficacy. An efficacy study in mouse using a GPX4-sensitive tumor model found that doses of 24up to 50 mg/kg were tolerated for 20 days but had no effect on tumor growth, although partial target engagement was observed in tumor homogenate.

Anti-fibrinolytic agents are used to reduce obstetric blood loss as the fibrinolytic system is known to get activated after placental delivery. To evaluate the efficacy of parenteral tranexamic acid in reducing blood loss during normal labour and to compare it with the amount of blood loss in patients who received placebo in the third stage of labour. Patients with spontaneous labour or planned for induction of labour and fulfilling the inclusion criteria were recruited for the study. In each patient, the pre-delivery pulse rate, blood pressure, Hb gm% and PCV% were noted. Labour was monitored carefully using a partogram. The study group received Inj. Oxytocin and Inj. Tranexamic acid. The control group received Inj. Oxytocin and Placebo injection. Immediately after delivery of the baby, when all the liquor was drained, the patient was placed over a blood drape—a disposable conical, graduated plastic collection bag. The amount of blood collected in the blood drape was measured. Then the patient was given pre-weighed pads, which were weighed 2 h post-partum. The blood loss was measured by measuring the blood collected in the drape and by weighing the swabs before and after delivery. The total number of patients studied was 100—equally distributed in both the groups. The age group of the patients and BMI were comparable. There was a significant increase in the pulse rate and decrease in blood pressure in the control group as compared with the study group. The post-delivery haemoglobin and haematocrit were significantly reduced in the control group as compared to the study group. The mean blood loss at the end of 2 h was 105 ml in the study group and 252 ml in the control group. There was a significant increase in the usage of uterotonics and also in the need for blood transfusion in the control group; 12 % of the patients in the control group had to stay for more than 3 days compared to 2 % in the study group. Tranexamic acid injection, an antifibrinolytic agent when given prophylactically after the delivery of the baby, by intravenous route appears to reduce the blood loss and maternal morbidity during normal labour effectively.

The third stage of labour commences after the delivery of the foetus and ends with the delivery of the placenta and its membranes. Postpartum haemorrhage is the most common cause of maternal mortality and accounts for about 25 % of maternal deaths in India. The present study was designed to evaluate the effectiveness of placental blood drainage after spontaneous vaginal delivery as part of active management of third stage of labour in decreasing the duration, blood loss, and complications of the third stage, against no drainage of placental blood. Two hundred pregnant patients with 37 or more weeks of gestation, with single live foetus in cephalic presentation, who underwent a spontaneous vaginal delivery, were included in the study. The patients were prospectively randomized equally into two groups (100 each in the study and control groups). Placental blood was drained in all the patients in the study group, whereas in the control group the cord blood was not drained. Blood lost in the third stage of labour was measured by collecting in a disposable conical measuring bag, and blood from the episiotomy was mopped, and the mops were discarded separately. The baseline statistics in both the group were comparable. The duration of third stage of labour was 210.5 s in the study group and 302.5 s in the control group. The ‘p’ value was statistically significant (p≤ 0.0001). The mean blood loss in study group was 227.5 ml and was 313.3 ml in the control group (p≤ 0.0001). The incidence of postpartum haemorrhage was 1 % in study group and 9 % in control group. The mean drop in Hb % level was 0.6 gm/dl in study group and 1.1 gm/dl in control group. These above differences were both statistically significant. Placental blood drainage as part of active management of third stage of labour was effective in reducing the duration, the blood loss, and also the incidence of PPH. Placental blood drainage is a simple, safe, and non-invasive method of managing the third stage of labour, which can be practiced in both tertiary care centres as well as rural setup in addition to the routine uterotonics.

The increased complications to the mother and fetus during or after pregnancy and birth are often caused by a wide array of pathogenic organisms mostly belonging to the TORCH group [toxoplasmosis, rubella, cytomegalovirus (CMV), and herpes simplex virus (HSV)]. These agents cause asymptomatic or mild infection in the mother while serious consequences in fetus. The present study was aimed to find significant etiological pathogens in the causation of high risk pregnancy (HRP) in South Indian population. A total of 1,158 HRP women (2010–2013) from Modern Government Maternity Hospital, Hyderabad were considered. Two milliliter of blood was obtained and the serum was analyzed for IgG and IgM antibodies against TORCH agents by ELISA. Twenty-five percent of the study group had fetal congenital malformation in the present pregnancy (Group 1; N= 291) while 75 % showed bad obstetric history (BOH) (Group 2; N= 867). Maternal age of ≤25 years, primi gravida, and consanguinity showed predisposing role for Group 1 while maternal age ≥30 years and ≥ 3 gravida were contributing risk for Group 2. The seropositvity in HRP women for toxoplasma, rubella, CMV, and HSV was 28, 84, 92, and 61 %, respectively for IgG while it was 6, 3, 4, and 3 % for IgG + IgM. Total seropositvity of toxoplasma, rubella, CMV, and HSV in Group 1 was 29, 97, 97, and 62 % while it was 36, 84, 97, and 65 % in Group 2, respectively. Maternal age of ≤25 years, primi gravida, and consanguinity contributed to fetal congenital malformation in the present pregnancy while maternal age of ≥30 years and ≥ 3 gravida towards BOH. Toxoplasma is protective while rubella and CMV are the infectious agents for HRP. Among the groups, toxoplasma and rubella conferred a predisposing risk towards Group 2 and Group 1, respectively. Sixty-one percent seropositvity of HSV in relation to bad obstetric outcome is the highest prevalence reported so far in India.

Due to depleting reserves of fossil fuels, political uncertainties, increase in demand of energy needs and growing concerns of environmental effects, bioenergy as an alternative source of energy needs had taken centre stage globally. In this report, we review the progress made in lignocellulose, cellulose and fermentation based biofuels in addition to tree borne oil seeds. Algae as a source of feedstock for the biofuel has also been reviewed. Recent efforts in genome sequencing of biofuel crops and molecular breeding approaches have increased our understanding towards crop improvement of major feedstocks. Besides, patenting trends in bioenergy sector were assessed by patent landscape analysis. The results showed an increasing trend in published patents during the last decade which is maximum during 2011. A conceptual framework of “transgenesis in biofuels to industrial application” was developed based on the patent analytics viz., International Patent Classification (IPC) analysis and Theme Maps. A detailed claim analysis based on the conceptual framework assessed the patenting trends that provided an exhaustive dimension of the technology. The study emphasizes the current thrust in bioenergy sector by various public and private institutions to expedite the process of biofuel production.

KCC2, potassium chloride cotransporter 2, is expressed exclusively in the CNS (on inhibitory neurons) and plays a major role in maintaining appropriately low intracellular chloride levels that ensure inhibitory actions of GABAAand glycine receptors. As such, it plays a pivotal role in inhibitory mechanisms that control neuronal excitation in the CNS. KCC2 downregulation has been implicated in various excitatory disorders, such as epilepsy and neuropathic pain. Positive modulators of KCC2 expression or activity may thus provide effective therapy for these disorders. However, the identification of such agents is hindered by the lack of a high-throughput screening method. Here the authors report the development of a fluorescence-based thallium (Tl+) transport assay using a Fluorometric Imaging Plate Reader (FLIPR), in which KCC2 activity is assessed by measuring the initial rate of KCC2-mediated Tl+transport/influx. The authors demonstrate Tl+/Cl–cotransport by KCC2, which exhibits a high apparent affinity for Tl+and dependency on the presence of the Cl–ion. Pharmacological studies revealed anticipated effects and potencies of known KCC-positive (NEM, staurosporine) and KCC-negative (DIOA, furosemide) modulators. The authors demonstrate that the assay is robust and reproducible and can be employed in high-throughput screening for positive modulators of KCC2 as potential therapeutic agents.

High throughput screening (HTS) has become a pivotal part of drug discovery for identifying potential lead molecules for specific disease targets. The success of screening is measured by several factors including the throughput and cost of the assays, but ultimately by the number of HTS hits that progress forward through the drug discovery pipeline. Miniaturization of assays by conversion from 96- to 384- to 1536- wells to well-less platforms, and the utilization of automated equipments are methodologies implemented to increase the throughput and reduce the cost of the screen. Pooling of test compounds is also an effective, though controversial method to enhance screening throughput while reducing reagent costs. Herein, we describe specific measures implemented at Abbott Laboratories to enhance the efficiency of 10- mixture orthogonal screening, and the strategy developed to reduce the rate of false positives and eliminate false negatives. Results from a comparison study of mixtures versus single compound screening will be discussed for three distinct screening formats: a GPCR functional assay, a GPCR binding assay and a tyrosine kinase assay.

Adventitious shoot regeneration from root, hypocotyl, cotyledon and primary leaf explants of safflower (Carthamus tinctoriusL.) was studied. Shoot regeneration was promoted by benzyladenine (BA) + naphthaleneacetic acid (NAA), BA + indole-3-butyric acid (IBA), kinetin + NAA and thidiazuron (TDZ) + NAA incorporated in Murashige and Skoog (MS) basal medium. High frequency of shoot regeneration and high number of shoots per regenerating explant were obtained on a wide range of TDZ + NAA combinations. Proliferated shoots were elongated in MS + 0.5 mg dm−3kinetin and well-developed shoots were rooted in half strength MS + 0.5 mg dm−3NAA. Rooted shoots were successfully acclimatized and established in soil.

Protocols for in vitro propagation of non-toxic variety of J. curcas through axillary bud proliferation and direct adventitious shoot bud regeneration from leaf segments have been established. Shoot bud proliferation from axillaries was assessed on an initial basal Murashige and Skoog (MS) salt medium supplemented with different concentrations of benzyladenine (BA), kinetin and thidiazuron (TDZ) followed by subculture to medium with 4.4-8.9  μM BA. Regardless of the concentration of BA in the subculture medium, shoot multiplication rate was optimum (10–12.3) with primary culture on medium supplemented with 2.3–4.5 μM TDZ. Efficient adventitious shoot regeneration from leaf tissues was achieved with culture on medium with 8.9–44.4 μM BA + 4.9 μM indole-3-butyric acid (IBA) followed by transfer to medium supplemented with 8.9 μM BA + 2.5 μM IBA. Similarity index between toxic Indian variety and the non-toxic variety based on 435 RAPD markers was 96.3%. Crossing studies followed by phorbol ester quantitation revealed that outcrosses with toxic J. curcas do not affect the phorbol ester content of seeds borne on the non-toxic variety.

Abstract A protocol for the transformation of castor embryo axes using the pCAMBIA vector 1304 in disarmed Agrobacterium tumefaciens strain EHA105 is presented. Co-cultivated explants were initially subjected to expansion and proliferation on MS medium with 0.5 mg l-1 TDZ followed by three cycles of selection on medium with 0.5 mg l-1 BA and increasing concentrations of hygromycin (20–40–60 mg l-1). Selected shoot clusters were transferred to medium with 0.5 mg l-1 BA for proliferation and 0.2 mg l-1 BA for shoot elongation. Elongated shoots were rooted on half-strength MS medium with 2.0 mg l-1 NAA. The presence and stable integration of the hpt gene was confirmed through PCR, RT-PCR, PCR-Southern blot, sequence analysis, Southern blot analysis and PCR analysis of progeny. Southern blot analysis of the primary transformants showed single copy integration and progeny analysis revealed monogenic inheritance of the introduced gene. This paper reports the first successful attempt at producing transgenic castor.

Although fluorescence imaging plate reader (FLIPR)-based assays have been widely used in high-throughput screening, improved efficiencies in throughput and fidelity continue to be investigated. This study presents an offline compound addition protocol coupled with a testing strategy using mixtures of compounds in a 384-well format to identify antagonists of the neurokinin-1 receptor expressed in the human astrocytoma cell line (U373 MG). Substance P evoked a concentration-dependent increase in intracellular cellular Ca2+with an EC50value of 0.30 ± 0.17 nM, which was inhibited by neurokinin-1 (NK1) antagonists L-733,060 and L-703,606. Test compounds, as mixtures of 10 compounds/well, were added to the cells offline using an automated dispensing unit and incubated prior to performing the assay in the FLIPR. Using the offline protocol, a higher through put of ~200,000 compounds was achieved in an 8-h working day, and several novel structural classes of compounds were identified as antagonists for the NK1 receptor. These studies demonstrate that the offline compound addition format using a mixture of compounds in a 384-well FLIPR assay provides an efficient platform for screening and identifying modulators for G-protein-coupled receptors. (Journal of Biomolecular Screening2005:46-55)

Interspecific hybridization has been successful between two economically important species of Jatropha, viz., J. curcasand J. integerrima. The interspecific hybrids exhibited morphological intermediacy for various vegetative characteristics but produced flowers with three distinct colours. Backcrossing of the F1hybrids resulted in a number of flower colours varying from dark pink through green to white enhancing the ornamental value of the genus. Hybridity was confirmed through PCR amplification using random primers. The various propagation methods for these new ornamental Jatrophasare discussed.

Micro Arrayed Compound Screening (pARCS) is a miniaturized ultra-high-throughput screening platform developed at Abbott Laboratories. In this format, 8640 discrete compounds are spotted and dried onto a polystyrene sheet, which has the same footprint as a 96-well plate. A homogeneous time-resolved fluorescence assay format (LANCE) was applied to identify the inhibitors of caspase-3 using a peptide substrate labeled with a fluorescent europium chelate and a dabcyl quencher. The caspase-3 enzyme was cast into a thin agarose gel, which was placed on a sheet containing test compounds. A second gel containing caspase substrate was then laid above the enzyme gel to initiate the reaction. Caspase-3 cleaves the substrate and separates the europium from the quencher, giving rise to a time-resolved fluorescent signal, which was detected using a ViewLux charge-coupled device imaging system. Potential inhibitors of caspase-3 appeared as dark spots on a bright fluorescent background. Results from the pARCS assay format were compared to those from a conventional 96-well plate-screening format.

Micro Arrayed Compound Screening (μARCS) is a miniaturized ultra-high-throughput screening platform developed at Abbott Laboratories. In this format, 8640 discrete compounds are spotted and dried onto a polystyrene sheet, which has the same footprint as a 96-well plate. A homogeneous time-resolved fluorescence assay format (LANCE) was applied to identify the inhibitors of caspase-3 using a peptide substrate labeled with a fluorescent europium chelate and a dabcyl quencher. The caspase-3 enzyme was cast into a thin agarose gel, which was placed on a sheet containing test compounds. A second gel containing caspase substrate was then laid above the enzyme gel to initiate the reaction. Caspase-3 cleaves the substrate and separates the europium from the quencher, giving rise to a time-resolved fluorescent signal, which was detected using a ViewLux charge-coupled device imaging system. Potential inhibitors of caspase-3 appeared as dark spots on a bright fluorescent background. Results from the μARCS assay format were compared to those from a conventional 96-well plate-screening format.

A fluorescence-based assay using the FLIPR Membrane Potential Assay Kit (FMP) was evaluated for functional characterization and high throughput screening (HTS) of potassium channel (ATP-sensitive Ki channel; KATP) modulators. The FMP dye permits a more sensitive evaluation of changes in membrane potential with a more rapid response time relative to DiBAC4(3). The time course of responses is comparable to ligand-evoked activation of the channel measured by patch-clamp studies. The pharmacological profile of the K+ channel evaluated by using reference KATP channel openers is in good agreement with that derived previously by DiBAC4(3)-based FLIPR assays. Improved sensitivity of responses together with the diminished susceptibility to artifacts such as those evoked by fluorescent compounds or quenching agents makes the FMP dye an alternative choice for HTS screening of potassium channel modulators.