Investigating the utility of satellite-based precipitation products for simulating extreme discharge events: an exhaustive model-driven approach for a tropical river basin in India.

Satellite-based precipitation estimates are a critical source of information for understanding and predicting hydrological processes at regional or global scales. Given the potential variability in the accuracy and reliability of these estimates, comprehensive performance assessments are essential before their application in specific hydrological contexts. In this study, six satellite-based precipitation products (SPPs), namely, CHIRPS, CMORPH, GSMaP, IMERG, MSWEP, and PERSIANN, were evaluated for their utility in hydrological modeling, specifically in simulating streamflow using the Variable Infiltration Capacity (VIC) model. The performance of the VIC model under varying flow conditions and timescales was assessed using statistical indicators, viz., R2, KGE, PBias, RMSE, and RSR. The findings of the study demonstrate the effectiveness of VIC model in simulating hydrological components and its applicability in evaluating the accuracy and reliability of SPPs. The SPPs were shown to be valuable for streamflow simulation at monthly and daily timescales, as confirmed by various performance measures. Moreover, the performance of SPPs for simulating extreme flow events (streamflow above 75%, 90%, and 95%) using the VIC model was assessed and a significant decrease in the performance was observed for high-flow events. Comparative analysis revealed the superiority of IMERG and CMORPH for streamflow simulation at daily timescale and high-flow conditions. In contrast, the performances of CHIRPS and PERSIANN were found to be poor. This study highlights the importance of thoroughly assessing the SPPs in modeling diverse flow conditions.

Nitric oxide mediated regulation of ascorbate-glutathione pathway alleviates mitotic aberrations and DNA damage in Allium cepa L. under salinity stress.

Allium cepa L. is an important medicinal and food plant enormously affected by salinity in terms of its growth and quality. This experiment investigates ameliorative potential of NO donor sodium nitroprusside (SNP) on chromosomal aberrations and physiological parameters in A. cepa L. roots exposed to salinity stress. Roots with different concentrations of NaCl (25, 50, and 100 mM) alone, and in combination with 100 µM SNP were analyzed for mitotic aberrations, DNA damage, proline, malondialdehyde (MDA) content, and ascorbate-glutathione (AsA-GSH) cycle after 120 h of salinity treatments. Results revealed that salinity stress increased chromosomal aberrations, MDA, proline accumulation, and severely hampered the AsA-GSH cycle function. The comet assay revealed a significant (p ≤ 0.05) enhancement in tail length (4.35 ± 0.05 µm) and olive tail moment (3.19 ± 0.04 µm) at 100 mM NaCl exposure. However, SNP supplementation decreased total percent abnormalities, while increased the prophase, metaphase, anaphase, and telophase indexes. Moreover, ascorbate peroxidase and glutathione reductase activities increased with AsA/DHA and GSH/GSSG ratios, respectively. Results suggest that SNP supplementation alleviates salinity stress responses by improving AsA-GSH cycle and proline accumulation. Based on present findings, NO supplementation could be recommended as a promising approach for sustainable crop production under salinity stress.

Allium cepa L. response to salt stress has been investigated but its role on chromosomal changes and DNA damage are less investigated therefore, our focus is to explore NO supplementation effects on cytological aberrations and biochemical responses in A. cepa L. roots under salinity stress.

Integrated analysis of long-noncoding RNA and circular RNA expression in Peste-des-Petits-Ruminants Virus (PPRV) infected marmoset B lymphocyte (B95a) cells.

Peste-des-Petits-Ruminants (PPR) or goat plague is an important viral disease of sheep and goats caused by the small ruminant morbilli virus or PPR virus (PPRV). Long non coding RNAs (lncRNA) and circular RNAs (circRNA) play a pivotal role in several biological processes including regulation of virus-host interactions. The present study explored the expression of lncRNA, circRNA and their functions in PPRV infected B-lymphocyte (B95a) cells. The results revealed a total of 4531 lncRNA and 2348 circRNA expression in both mock and PPRV infected samples. Analysis of differentially expressed (DE) RNA identified 123 DE-lncRNA and 39 DE-circRNA as significantly dysregulated. Functional analysis of cis-target genes of DE-lncRNA indicated activation of TCF dependent WNT signaling and PKN1 stimulated transcription process. Interactions (sponging) of microRNA (miRNA) revealed 344 DE-lncRNA-miRNA and 93 DE-circRNA-miRNA pairs. The competing endogenous RNA (ceRNA) network of lncRNA/circRNA-miRNA-mRNA in PPRV infected B95a cells was represented by 69 ceRNA pairs. We validated the DE-circRNA by targeted amplification and sequencing of back spliced junctions (BSJs). The present study revealed a profile of lncRNA, circRNA and their potential ceRNA network in PPRV infection. The results provide insight for better understanding of PPRV-host interactions.

Effect of acute heat shock on stress gene expression and DNA methylation in zebu (Bos indicus) and crossbred (Bos indicus × Bos taurus) dairy cattle.

Environmental temperature is one of the major factors to affect health and productivity of dairy cattle. Gene expression networks within the cells and tissues coordinate stress response, metabolism, and milk production in dairy cattle. Epigenetic DNA methylations were found to mediate the effect of environment by regulating gene expression patterns. In the present study, we compared three Indian native zebu cattle, Bos indicus (Sahiwal, Tharparkar, and Hariana) and one crossbred Bos indicus × Bos taurus (Vrindavani) for stress gene expression and differences in the DNA methylation patterns. The results indicated acute heat shock to cultured PBMC affected their proliferation, stress gene expression, and DNA methylation. Interestingly, expressions of HSP70, HSP90, and STIP1 were found more pronounced in zebu cattle than the crossbred cattle. However, no significant changes were observed in global DNA methylation due to acute heat shock, even though variations were observed in the expression patterns of DNA methyltransferases (DNMT1, DNMT3a) and demethylases (TET1, TET2, and TET3) genes. The treatment 5-AzaC (5-azacitidine) that inhibit DNA methylation in proliferating PBMC caused significant increase in heat shock-induced HSP70 and STIP1 expression indicating that hypomethylation facilitated stress gene expression. Further targeted analysis DNA methylation in the promoter regions revealed no significant differences for HSP70, HSP90, and STIP1. However, there was a significant hypomethylation for BDNF in both zebu and crossbred cattle. Similarly, NR3C1 promoter region showed hypomethylation alone in crossbred cattle. Overall, the results indicated that tropically adapted zebu cattle had comparatively higher expression of stress genes than the crossbred cattle. Furthermore, DNA methylation may play a role in regulating expression of certain genes involved in stress response pathways.

Deriving wetland-cover types (WCTs) from integration of multispectral indices based on Earth observation data.

The wetland cover is defined as the spatially homogenous region of a wetland attributed to the underlying biophysical conditions such as vegetation, turbidity, hydric soil, and the amount of water. Here, we present a novel method to derive the wetland-cover types (WCTs) combining three commonly used multispectral indices, NDVI, MNDWI, and NDTI, in three large Ramsar wetlands located in different geomorphic and climatic settings across India. These wetlands include the Kaabar Tal, a floodplain wetland in east Ganga Plains, Chilika Lagoon, a coastal wetland in eastern India, and Nal Sarovar in semi-arid western India. The novelty of our approach is that the derived WCTs are stable in space and time, and therefore, a given WCT across different wetlands or within different zones of a large wetland will imply similar underlying biophysical attributes. The WCTs can therefore provide a novel tool for monitoring and change detection of wetland cover types. We have automated the proposed WCT algorithm using the Google Earth Engine (GEE) environment and by developing ArcGIS tools. The method can be implemented on any wetland and using any multispectral imagery dataset with visible and NIR bands. The proposed methodology is simple yet robust and easy to implement and, therefore, holds significant importance in wetland monitoring and management.

Penumbra quantification from MR SWI-DWI mismatch and its comparison with MR ASL PWI-DWI mismatch in patients with acute ischemic stroke.

In acute-ischemic-stroke patients, penumbra assessment plays a significant role in treatment outcome. MR perfusion-weighted imaging (PWI) and diffusion-weighted imaging (DWI) mismatch ratio can provide penumbra assessment. Recently reported studies have shown the potential of susceptibility-weighted imaging (SWI) in the qualitative assessment of penumbra. We hypothesize that quantitative penumbra assessment using SWI-DWI can provide an alternative to the PWI-DWI approach and this can also reduce the overall scan-time. The purpose of the current study was to develop a framework for accurate quantitative assessment of penumbra using SWI-DWI and its validation with PWI-DWI-based quantification. In the current study, the arterial-spin-labelling (ASL) technique has been used for PWI. This retrospective study included 25 acute-ischemic-stroke patients presenting within 24 hours of the last noted baseline condition of stroke onset. Eleven patients also had follow-up MRI within 48 hours. MRI acquisition comprised DWI, SWI, pseudo-continuous-ASL (pCASL), FLAIR and non-contrast-angiography sequences. A framework was developed for the enhancement of prominent hypo-intense vein signs followed by automatic segmentation of the SWI penumbra ROI. Apparent-diffusion-coefficient (ADC) maps and cerebral-blood-flow (CBF) maps were computed. The infarct core ROI from the ADC map and the ASL penumbra ROI from CBF maps were segmented semiautomatically. The infarct core volume, SWI penumbra volume (SPV) and pCASL penumbra volume were computed and used to calculate mismatch ratios MRSWIADC and MRCBFADC . The Dice coefficient between the SWI penumbra ROI and ASL penumbra ROI was 0.96 ± 0.07. MRSWIADC correlated well (r = 0.90, p < 0.05) with MRCBFADC , which validates the hypothesis of accurate penumbra assessment using the SWI-DWI mismatch ratio. Moreover, a significant association between high SPV and the presence of vessel occlusion in the MR angiogram was observed. Follow-up data showed salvation of penumbra tissue (location and volumes predicted by proposed framework) by treatments. Additionally, functional-outcome analysis revealed 93.3% of patients with MRSWIADC > 1 benefitted from revascularization therapy. Overall, the proposed automated quantitative assessment of penumbra using the SWI-DWI mismatch ratio performs equivalently to the ASL PWI-DWI mismatch ratio. This approach provides an alternative to the perfusion sequence required for penumbra assessment, which can reduce scan time by 17% for the protocol without a perfusion sequence.

Replication competence of canine distemper virus in cell lines expressing signaling lymphocyte activation molecule (SLAM) of goat, sheep and dog origin.

Cellular receptors play an important role in entry and cell to cell spread of morbillivirus infections. The cells expressing SLAM and Nectin-4 have been used for successful and efficient isolation of canine distemper virus (CDV) in high titre. There are several methods for generation of cells expressing receptor molecules. Here, we have used a comparatively cheaper and easily available method, pcDNA 3.1 (+) for engineering Vero cells to express SLAM gene of goat, sheep and dog origin (Vero/Goat/SLAM (VGS), Vero/Sheep/SLAM (VSS) and Vero/Dog/SLAM (VDS), respectively). The generated cell lines were then compared to test their efficacy to support CDV replication. CDV could be grown in high titre in the cells expressing SLAM and a difference of log two could be recorded in virus titre between VDS and native Vero cells. Also, CDV could be grown in a higher titre in VDS as compared to VGS and VSS. The finding of this study supports the preferential use of SLAM expressing cells over the native Vero cells by CDV. Further, the higher titre of CDV in cells expressing dog-SLAM as compared to the cells expressing SLAM of non-CDV hosts (i.e. goat and sheep) points towards the preferential use of dog SLAM by the CDV and may be a plausible reason for differential susceptibility of small ruminants and Canines to CDV.

Development of novel iron-regulated Pasteurella multocida B: 2 bacterin and refinement of vaccine quality in terms of minimum variation in particle size and distribution vis-a-vis critical level of iron in media.

To enhance the qualitative bacterial biomass per unit of media and to overcome the limitations of the existing haemorrhagic septicaemia (HS) vaccines, a comprehensive study was undertaken encompassing the role of iron on the bacterial biomass of Pasteurella multocida B: 2 to vaccine development. Trypsin digested hydrochloric acid-treated sheep blood (THSB) as a novel iron rich supplement had been devised for the first time for augmenting the qualitative bacterial biomass per unit of media which was evident with growth kinetic study. The higher recovery of iron from THSB became evident via atomic absorbance spectrophotometry. The critical level of iron in the media as well as mode of iron supplementation showed a major impact on the outer membrane protein profile of P. multocida B:2 and variation in droplet size and particle-size distribution of formulated vaccine. Immune response study against iron-regulated bacterin adjuvanted with aluminum hydroxide gel in mouse model showed that 3% THSB supplementation of casein sucrose yeast (CSY) not only augmented the growth of P. multocida B:2 significantly but conferred highest pre-challenged ELISA IgG titer and protection against pasteurellosis. Thus, THSB supplementation of CSY can resolve existing up-scaling and immunogenic potential problems of HS vaccine production.

Interactive role of exogenous 24 Epibrassinolide and endogenous NO in Brassica juncea L. under salinity stress: Evidence for NR-dependent NO biosynthesis.

The present study unravels origin of nitric oxide (NO) and the interaction between 24-Epibrassinolide (EBL) and nitrate reductase (NR) for NO production in Indian mustard (Brassica juncea L.) under salinity stress. Two independent experiments were performed to check whether (i) Nitrate reductase or Nitric oxide synthase takes part in the biosynthesis of endogenous NO and (ii) EBL has any regulatory effect on NR-dependent NO biosynthesis in the alleviation of salinity stress. Results revealed that NR-inhibitor tungstate significantly (P ≤ 0.05) decreased the NR activity and endogenous NO content, while NOS inhibitor l-NAME did not influence NO biosynthesis and plant growth. Under salinity stress, inhibition in NR activity decreased the activities of antioxidant enzymes, increased H2O2, MDA, protein carbonyl content and caused DNA damage, implying that antioxidant defense might be related to NO signal. EBL supplementation enhanced the NR activity but did not influence NOS activity, suggesting that NR was involved in endogenous NO production. EBL supplementation alleviated the inhibitory effects of salinity stress and improved the plant growth by enhancing nutrients, photosynthetic pigments, compatible osmolytes, and performance of AsA-GSH cycle. It also decreased the superoxide ion accumulation, leaf epidermal damages, cell death, DNA damage, and ABA content. Comet assay revealed significant (P ≤ 0.05) enhancement in tail length and olive tail moment, while flow cytometry did not showed any significant (P ≤ 0.05) changes in genome size and ploidy level under salinity stress. Moreover, EBL supplementation increased the G6PDH activity and S-nitrosothiol content which further boosted the antioxidant responses under salinity stress. Taken together, these results suggested that NO production in mustard occurred in NR-dependent manner and EBL in association with endogenous NO activates the antioxidant system to counter salinity stress.

Lentiviral-mediated delivery of classical swine fever virus Erns gene into porcine kidney-15 cells for production of recombinant ELISA diagnostic antigen.

Classical swine fever virus (CSFV), a member of the Pestivirus genus within the Flaviviridae family causes contagious fatal disease in swine. Antibodies against E2, Erns and NS3 proteins of virus can be detected in infected animals. Development of an ELISA coating antigen to improve the sensitivity of detecting Erns-specific antibodies in pig sera is always desirable for diagnosis as well as for differentiation of infected from vaccinated animals. In present study, a lentivirus-based gene delivery system was used to develop a stable PK-15 cell line expressing Erns (PK-Erns) for production of diagnostic antigen. The Lenti-Erns virus was purified from the supernatant of co-transfected 293LTV cells and used to transduce PK-15 cells. The homogenous PK-Erns cell line was produced by single cell cloning by monitoring eGFP expression. The Erns gene in the genomic DNA and RNA transcripts in total RNA isolated from PK-Erns cells were detected by PCR and RT-PCR, respectively. Expression of 45 kDa Erns glycoprotein was detected in western blot using CSFV-specific hyperimmune sera. The use of PK-Erns cell lysate as antigen in serial dilution and single dilution ELISAs with known positive and negative pig sera was investigated. The PK-Erns ELISA revealed sensitivity equivalent to commercial HerdChek ELISA kit. The sensitivity, specificity and accuracy of the PK-Erns ELISA was 95%, 100% and 96.66%, respectively compared to ELISA using purified CSFV as coating antigen. When field pig sera (n = 69) were tested in PK-Erns ELISA, a significant correlation between the titers from serial dilution and single dilution ELISA was observed. This indicated that PK-Erns cell line can serve as continuous source of ELISA diagnostic antigen for detection of CSFV-specific antibodies in pig sera.

3D-Double-Inversion recovery detects perilesional gliosis better than 3D-FLAIR and postcontrast T1 imaging in calcified neurocysticercosis.

BACKGROUND: Perilesional gliosis is an important substrate for seizures in patients harboring a calcified neurocysticercosis (NCC) lesion and magnetic resonance imaging (MRI) is useful for evaluating gliosis. AIMS: The purpose of this study was to evaluate the usefulness of double-inversion recovery (DIR) sequence for identifying perilesional gliosis. SETTINGS AND DESIGN: Hospital-based cross-sectional study. METHODS AND MATERIALS: Forty-five patients with seizures were included in this study and a total of 88 calcified lesions identified on susceptibility weighted imaging (SWI) were evaluated on 3D-fluid attenuating inversion recovery (FLAIR), 3D-DIR, and 3D-postcontrast T1-weighted imaging on a 3T MRI for the presence of perilesional signal changes/enhancement. Perilesional signal was rated on a semiquantitative scale from grade 0 to 2 by independent raters. STATISTICAL ANALYSIS USED: Friedman, Wilcoxon signed rank, and Kappa tests were used. RESULTS: 3D-DIR sequence performed better than both 3D-FLAIR and postcontrast 3D-T1W sequences as more number of lesions showed perilesional signal change on DIR sequence. DIR sequence showed perilesional signal abnormality in 24 lesions in which 3D-FLAIR was normal, whereas in another 18 lesions, it demonstrated perilesional signal changes better than 3D-FLAIR. In only three lesions, FLAIR was found to be superior to DIR sequence, whereas postcontrast T1W images showed rim enhancement in five cases where no perilesional signal change was seen on FLAIR/DIR sequences. CONCLUSIONS: Combining 3D-DIR with 3D-FLAIR, and postcontrast 3D-T1W sequences is beneficial for evaluation of calcified NCC lesions and 3D-DIR sequence is better than other two sequences for perilesional signal abnormalities.

Counteractive mechanism (s) of salicylic acid in response to lead toxicity in Brassica juncea (L.) Czern. cv. Varuna.

MAIN CONCLUSION: Salicylic acid alleviates lead toxicity in Brassica juncea (L.) by promoting growth under non-stress and activating stress-defense mechanism (s) under lead stress conditions. It also boosts the ascorbate-glutathione cycle and thus helps in minimizing oxidative and DNA damage. Brassica juncea plants were exposed to different concentrations (0, 500, 1000 and 2000 mg kg-1) of lead (Pb) and subsequently sprayed with 0.5 mM of salicylic acid (SA) to check for morphological and leaf gas exchange parameters like transpiration rate (E), stomatal conductance (GH2O), net photosynthetic rate (A) and maximum quantum yield of PS II (Fv/Fm). Leaf epidermis by scanning electron microscopy (SEM), enzymatic and non-enzymatic components of ascorbate-glutathione (AsA-GSH) cycle, DNA damage by comet assay, lipid peroxidation and endogenous SA quantification by HPLC were analyzed. Lead accumulation in root, shoot and its sub-cellular distribution ratio (SDR) and localization was also determined using atomic absorption spectroscopy (AAS) and rhodizonate-dye staining method, respectively. Results revealed that notable amount of Pb was accumulated in root and shoot in dose-dependent manner which significantly (P ≤ 0.05) posed the toxicity on the majority of morphological parameters, structural integrity of epidermal and guard cells, photosynthetic pigments, malondialdehyde (MDA) and H2O2 content. Notable decrease in leaf gas exchange parameters, Fv/Fm, poor performance of AsA-GSH cycle and striking amount of DNA damage, was found as well. However, SA revoked Pb toxicity to a great extent by promoting growth, chlorophyll content, improving the A, Fv/Fm, boosting the overall performance of AsA-GSH cycle and by lessening the DNA damage.

EvaGreen-based Multiplex Real-time PCR Assay for Rapid Differentiation of Wild-Type and Glycoprotein E-Deleted Bovine Herpesvirus-1 Strains.

Bovine herpesvirus-1 (BoHV-1) is an important viral pathogen causing significant economic losses to the cattle industry. Glycoprotein E-deleted marker vaccines form the basis for BoHV-1 control programs widely, wherein detection and differentiation of wild-type and gE-deleted vaccine strains is of crucial importance for proper disease management. In the present study, we report an EvaGreen-based multiplex real-time polymerase chain reaction (EGRT-PCR) assay for rapid differentiation of wild-type and glycoprotein E-deleted strains of BoHV-1. The EGRT-PCR assay could simultaneously detect two viral genes (glycoprotein B and E) and an internal positive control gene (bovine growth hormone- bGH), in a single-tube reaction. The analytical sensitivity of the EGRT-PCR assay was as little as 10 copies of the BoHV-1 DNA per reaction. The modified real-time PCR assay could successfully differentiate wild-type and gE-deleted BoHV-1 strains based on gene specific melting temperatures (Tm) peaks. Our results have shown that the EGRT-PCR developed in this study might prove to be a promising tool in disease management by enabling rapid differentiation of wild-type and gE-deleted strains of BoHV-1.

Visual Detection of Brucella spp. in Spiked Bovine Semen Using Loop-Mediated Isothermal Amplification (LAMP) Assay.

Several pathogens including Brucella spp. are shed in semen of infected bulls and can be transmitted to cows through contaminated semen during artificial insemination. The present study reports omp2a and bcsp31 gene based loop-mediated isothermal amplification (LAMP) assays for detection of Brucella genomic DNA in semen from infected bulls. The positive results could be interpreted visually by change in colour of reaction mixture containing hydroxyl naphthol blue (HNB) dye from violet to sky blue. LAMP assays based on omp2a and bcsp31 could detect as little as 10 and 100 fg of B. abortus S19 genomic DNA, respectively. Sensitivity of omp2a and bcsp31 LAMP assays for direct detection of organisms in bovine semen was 2.28 × 10(1) CFU and 2.28 × 10(2) CFU of B. abortus S19 in spiked bovine semen, respectively. The omp2a LAMP assay was found equally sensitive to TaqMan probe based real-time PCR and 100 times more sensitive than conventional PCR in identifying Brucella in spiked semen. The diagnostic applicability of the omp2a LAMP assay was evaluated with seventy-nine bovine semen samples and results were re-evaluated through TaqMan probe based real-time PCR and conventional PCR. Taken together, the omp2a LAMP assay is easy to perform, rapid and sensitive in diagnosis of Brucella spp. in bovine semen.

Loop-Mediated Isothermal Amplification for Rapid Detection and Differentiation of Wild-Type Bovine Herpesvirus-1 and Glycoprotein E-Deleted Marker Vaccine Strain.

Bovine herpesvirus-1 (BoHV-1) is an important viral pathogen affecting cattle and causing numerous reproductive disorders leading to significant economic losses to the cattle industry. The control programs for BoHV-1 are widely based on the use of glycoprotein E-deleted marker vaccines, wherein detection and differentiation of wild-type and gE-deleted vaccine strains is of crucial importance for proper disease management. In this study, we report rapid and simple loop-mediated isothermal amplification (LAMP) assays for detection and differentiation of gE-deleted BoHV-1 from wild-type virus under isothermal conditions. The assays could be completed in 90 mintes, including viral DNA isolation, target amplification and visual interpretation of results with naked eye. The analytical sensitivity of the assays was 10 times higher than conventional PCR and could detect as little as 100 fg of viral DNA per reaction. The applicability of LAMP for detection of BoHV-1 in bovine semen was assessed by testing semen samples collected from breeding bulls and compared with TaqMan real-time PCR (as gold standard). The LAMP assays had diagnostic specificity of 100%. The diagnostic sensitivity was 88.2% and 83.3% for gB- and gE-LAMP, respectively, when compared with TaqMan real-time PCR. Our results have shown that the LAMP method developed in this study is a potential tool for rapid, sensitive, specific, cost-effective, and user-friendly detection and differentiation of wild type BoHV-1 from gE-deleted marker vaccine.

Prokaryotic expression of ovis aries conglutinin encoding neck and carbohydrate recognition domain and its functional characterization.

Conglutinin, a soluble pattern recognition receptor of innate immune system in bovines is known for its potential defensive activity against microorganisms either by direct agglutination in the presence of calcium or by acting as opsonin. In the present study, sheep (Ovis aries) conglutinin encoding neck and carbohydrate recognition domain (rSCGN) was expressed in the E coli BL21 expression host. The recombinant conglutinin revealed molecular weight of 27 kDa in SDS PAGE and also in western blotting using antibuffalo conglutinin polyclonal serum. The protein was characterized further for its functional activity in various assays. In ELISA based sugar and LPS binding assay, the rSCGN revealed its high binding activity toward N-acetyl glucosamine and E. coli LPS in the presence and the absence of calcium ions, respectively. Hemagglutination of chicken red blood cells caused by Newcastle disease virus was not inhibited in the presence of rSCGN as it lacked complete collagenous region present in the native protein. In virus neutralization test, the recombinant protein was found to reduce multiplication of bovine herpes virus-1 propagated in MDBK cells. This prokaryotically expressed 27 kDa recombinant sheep conglutinin can serve as antigen in future studies to develop sandwich ELISA for assessing the level of native conglutinin in sheep serum.

Design, synthesis and pharmacological evaluation of novel polycyclic heteroarene ethers as PDE10A inhibitors: part II.

We report the design and synthesis of novel pyrrolo[3,2-b]quinoline containing heteroarene ethers as PDE10A inhibitors with good to excellent potency, selectivity and metabolic stability. Further optimization of this primary series resulted in the identification of 1-methyl-3-(4-{[3-(pyridine-4-yl)pyrazin-2-yl]oxy}phenyl)-1H-pyrrolo[3,2-b]pyridine 13a with good hPDE10A potency (IC50: 6.3 nM), excellent selectivity over other related PDEs and desirable physicochemical properties. The compound exhibited high peripheral and adequate brain levels upon oral dosing in rodents. The compound also showed excellent efficacy in multiple preclinical animal models related to psychiatric disorders, particularly schizophrenia.

Rapid detection of bovine herpesvirus 1 in bovine semen by loop-mediated isothermal amplification (LAMP) assay.

Bovine herpesvirus 1 (BoHV-1) is the most common viral pathogen found in bovine semen, causing numerous reproductive disorders leading to economic losses to the cattle industry. For rapid detection of BoHV-1 in bovine semen, in this study, we applied a loop-mediated isothermal amplification (LAMP) assay. The assay could be completed within 90 min, including total DNA isolation, target amplification, and visual interpretation of positive or negative results with the naked eye. The assay detected as little as 10 fg of BoHV-1 DNA per reaction. The analytical sensitivity of the assay was 0.2 TCID50 BoHV-1 per reaction, which was 100 times more sensitive than conventional PCR and comparable to TaqMan real-time PCR. The applicability of the assay was assessed by analysing 118 semen samples collected from breeding bulls. On comparison with TaqMan real-time PCR, the LAMP assay had a diagnostic sensitivity of 97 %, specificity of 100 %, and accuracy of 99.2 % for detection of BoHV-1 in bovine semen. The LAMP assay developed in this study is a rapid, sensitive, and cost-effective alternative for detection of BoHV-1 in bovine semen.