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• Computational and experimental work has been performed on cocrystal of hydrochlorothiazide (HCTZ) and nicotinamide (NCT). • H-bonding interaction was studied by using monomeric, dimeric and trimeric models of HCTZ-NCT cocrystal. • NH 2 and C = O groups were involved in neighbouring hydrogen bonding interactions. • QTAIM and NBO analysis were done to explore inter and intramolecular H-bond interactions. • MEPS was performed to observe the reactive sites of the molecules. The present work focuses on the study of structural, vibrational, chemical properties and the effect of hydrogen bonding interaction in the formation of a cocrystal of hydrochlorothiazide (HCTZ, a diuretic drug) with nicotinamide (NCT) using vibrational spectroscopy (FT-IR and FT-Raman) and density functional theory. HCTZ being a BCS class IV drug, has low permeability and solubility; therefore, its cocrystal is studied. Monomer, dimer and trimer of HCTZ-NCT cocrystal were studied to understand hydrogen bonding interactions. The shifting in the wavenumber and the elongation in bond lengths of the amide (NH 2) group of HCTZ and carbonyl (C = O) group of NCT were observed in all the models of HCTZ-NCT cocrystal due to the hydrogen bonding interaction. The importance of hydrogen bonding were also explored by natural bond orbital (NBO) analysis in support of the quantum theory of atom in molecules (QTAIM). The chemical reactivity of HCTZ, NCT and HCTZ-NCT cocrystal was examined by MEPS map, FMOs, global electronic reactivity descriptor, ECT descriptors, and molar refractivity. The lesser value of the HOMO-LUMO energy gap in the cocrystal than API reflects that the chemical reactivity of the cocrystal was better than API itself. The comparison between API(HCTZ) and cocrystal (HCTZ-NCT) implies that pharmaceutical cocrystals may be a better option to improve and enhance the pharmacological properties of drugs. [Display omitted] [ABSTRACT FROM AUTHOR]

[Display omitted] • Optimized structure monomer and dimer structures of A10 DLC using DFT. • Microscopic molecular dynamics were revealed through FT-IR, FT-Raman, and DFT. • Predicted the mesomorphic models of nematic and columnar of A10. • QTAIM analysis predicts the strength and nature of hydrogen bonding. Utilizing temperature-dependent Fourier Transform Infrared (FT-IR) and Fourier Transform Raman (FT-Raman) spectroscopies, we delved into the spectral properties of 4-((2,3,4-tris(decyloxy)phenyl)diazenyl)benzoic acid, a fascinating discotic liquid crystal (DLC) named A10. Our investigation unravels valuable insights into conformational dynamics, intermolecular hydrogen bonding, and structural stability, throughout enantiotropic phase transitions (Isotropic (Iso) → Discotic Nematic (N D) → Columnar (Col) → Crystalline (Cry)) during cooling. To further understand the mesogen's phasmid structure at atomic level, density functional theory (DFT) is employed, utilizing the B3LYP/6-3116G(d,p) method for optimization and frequency calculations of monomer and dimer of A10. The alignment of computationally predicted molecular models with observed vibrational spectra at room temperature affirms the precision of our theoretical model. Innovatively, our study has uncovered the potential formation of dimer assemblies, demonstrating the persistence of intermolecular hydrogen bonding within these dimers throughout each phase transition of the DLC. Furthermore, we have identified the strength of both intermolecular and intramolecular hydrogen bonding. These findings expand our knowledge of this unique material and pave the way for further research in the field of liquid crystals. [ABSTRACT FROM AUTHOR]

[Display omitted] • Monomeric and dimeric structure of discotic liquid crystal (DLC) A8 is optimized using density functional theory (DFT). • Phase transition of DLC A8 (Isotropic → Discotic nematic → Columnar → Crystalline) have been analysed with the help of temperature dependent FT-IR and FT- Raman spectra. • Combination of experimental (FT-IR and FT-Raman) and computational results allow us to elucidate the dynamics of the molecules during the phase transitions. • AIM analysis has predicted the strength and nature of the inter and intra molecular hydrogen bonding in DLCs. The discotic liquid crystal 4-((2, 3, 4-tris (octyloxy) phenyl) diazenyl) benzoic acid , hereafter referred as DLC A8, exhibited in dimeric form has been studied using a combination of quantum chemical approaches and vibrational spectroscopy. This study investigates the structural alteration of DLC A8 associated with phase transition. The phase transitions of DLC A8 are Iso → Discotic nematic → Columnar → Crystalline, which have been investigated using differential scanning calorimetry (DSC) accompanied with polarized optical microscopy (POM). Monotropic columnar mesophase was observed during the cooling cycle while discotic nematic mesophase was observed in both the heating and cooling cycles. Density functional theory (DFT) along with IR and Raman spectroscopic techniques were utilized to study the dynamics of molecules during phase transition. To predict the most stable conformation of the molecule, one-dimensional PES scans have been performed along 31 flexible bonds using DFT/B3LYP/6-311G++(d,p) method. Vibrational normal modes were analyzed in detail, taking potential energy contribution into account. The spectral analysis of FT-IR and FT-Raman was done by deconvoluting the structural sensitive bands. The agreement between the calculated IR and Raman spectra and the observed FT-IR and Raman spectra at room temperature confirms our theoretically predicted molecular model of investigated discotic liquid crystal. Moreover, our studies have unraveled the existence of intact intermolecular H-bonding of dimers throughout the phase transitions. [ABSTRACT FROM AUTHOR]

The present study was taken up to find out Beta 2 Microglobulin (β2M) gene variants and their association with IgG concentrations in colostrum of Murrah buffaloes. 40 newly parturated Murrah buffaloes dams were included in the present study. Polymerase chain reaction- single stranded conformational polymorphism (PCR-SSCP) technique was used to explore the polymorphism in β2M gene of buffaloes. The PCR-SSCP analysis and sequencing of different patterns obtained by silver staining of fragments pertaining to exon 1 through 4 and partial introns 1 through 3, revealed 9 SNPs which were further used to prepare the haplotypes. 7 different haplotypes were observed. Indirect ELISA was done to estimate IgG concentrations in colostrum (collected 1 hour before first milking) obtained from buffaloes. IgG levels in colostrum ranged from 11.22 to 185.1 mg/ml with a mean IgG concentration of 51.71 ± 5.99 mg/ml. The effect of different haplotypes was evaluated on IgG concentrations. The least square analysis of variance could not exhibit a significant effect of buffalo dam haplotype on colostral IgG concentration. [ABSTRACT FROM AUTHOR]

[Display omitted] • Computational and experimental work has been performed on hydrochlorothiazide (HCTZ). • H-bonding interaction was studied using monomeric, dimeric and trimeric models. • NH 2 and O S O groups were involved in neighboring interactions. • QTAIM and NBO analysis was done to explore inter and intramolecular H-bond interactions. Hydrochlorothiazide (HCTZ) being a diuretic drug widely used in anti-hypertensive therapy as it lowers the blood pressure by reducing the reabsorption of electrolytes in kidney resulting an increment of urine output and lowering the blood pressure. The purpose of the present work is to study the structural, vibrational and chemical properties of HCTZ based on its monomeric, dimeric and trimeric models by utilizing computational methods and experimental techniques. Density functional theory (DFT) with functional B3LYP and 6-311++G (d, p) basis set was used for a detailed computational study. Monomeric, dimeric and trimeric models of HCTZ have been studied for a better understanding of inter- and intramolecular hydrogen bonding. FT-IR (400–3800 cm−1) and FT-Raman (100–3600 cm−1) spectroscopy have been utilized for the characterization of HCTZ. The shifting in wavenumber of NH 2 and O S O group were observed in dimer and trimer due to the formation of intermolecular hydrogen bonding. Quantum theory of atoms in molecules (QTAIM) along with natural bond orbital (NBO) analysis were performed to examine the nature and strength of hydrogen bonding which showed that all the interactions were medium and partially covalent in nature; transition from LP(3)O15 → σ*(H46 → N32) and LP(3)O39 → σ*(H74 → N51) were responsible for the formation of O15•••H46 and O39•••H74 H-bonds, respectively. HOMO-LUMO energies predicted the chemical reactivity and stability of the molecule and the energy gap for dimer (4.6240 eV) and trimer (4.0493 eV) was found to be lesser than the monomer (5.0888 eV) which showed that the dimer and trimer have predicted more chemical reactivity in comparison to monomer. The most electronegative electrostatic potential was observed around the O S O group and the most electropositive potential around the amide group from MEPS analysis. Global as well as local reactivity descriptors have predicted the reactivity and hence, stability of the title molecule. [ABSTRACT FROM AUTHOR]

The present study was taken up to find out SNPs in Beta 2 Microglobulin (β2M) gene and their possible association with serum IgG concentrations of newborn buffalo calves. Forty newborn Murrah buffaloes calves were included in the present study. Polymerase chain reaction-single stranded conformational polymorphism (PCR-SSCP) technique was used to explore the polymorphism in β2M gene of the buffalo calves. The PCR-SSCP analysis and sequencing of different patterns obtained by silver staining of various exonic and intronic fragments revealed nine SNPs. All the identified SNPs were used to prepare the haplotypes for the calves. Fourteen different haplotypes were observed. Indirect ELISA was done to estimate IgG concentrations in serum samples of the calves. IgG levels in calf serum ranged from 0.25 to 14.88 mg/ml and mean IgG concentration in serum was estimated as 11.68 ± 0.75 mg/ml. The least square analysis of variance could not exhibit a significant effect of calf haplotype on serum IgG concentration. [ABSTRACT FROM AUTHOR]

Contents: Transition proteins (TNPs) are essential in chromatin condensation during spermiogenesis, and hence, they are the candidate genes for identifying sperm motility markers. Coding and in silico predicted promoter regions of these genes were investigated in crossbred and purebred cattle, and also, their mRNA quantification was done to explore its use as a diagnostic tool of infertility. PCR‐SSCP analysis revealed two band patterns in fragment III of TNP1 and fragment II of TNP2 gene. Sequence analysis revealed a deletion of "G" nucleotide in 3′UTR region of TNP1 and C>T SNP in intronic region of TNP2 gene. Least square analysis of variance did not reveal any significant influence of nucleotide deletion on any sperm motility parameters in both crossbred and purebred cattle. However, C>T SNP had a significant effect on initial progressive motility (p < 0.05) in purebred cattle and post‐thaw motility in overall cattle population. RT‐qPCR analysis did not reveal any significant variation in TNP1 and TNP2 gene expression among poorly motile and good quality spermatozoa of Vrindavani bulls. [ABSTRACT FROM AUTHOR]

The main objective of this study was to develop baseline topographical data of landmarks for locating liver and spleen and to generate sonographic calliper measurements along with dynamic parameters of these organs. Abdominal ultrasonography was performed on 50 native adult (Spiti horses and Himalayan hill mules) healthy unsedated equines of either sex. The thickness, character and echo-architecture of liver and spleen were recorded. In horses, the right lobe of liver was found from 8th to 15th intercostal space (ICS) and left lobe from 6th to 11th (±1) ICS, while in mules the right liver lobe was found between 6th to 15th (±1) ICS and left lobe between 6th to 9th (±1) ICS. The liver was recognized by its branching vasculature and architecture was found relatively homogenous. In addition ventral edges were distinctly sharp. Spleen was found from 8th to 17th ICS both in horses as well as in mules. The splenic parenchyma was homogenously granular in appearance with few blood vessels and most echogenic organ in the abdominal cavity of equines. The only measurement that was reliably obtained was the central thickness or depth of the spleen, which varied from 41.4 to 68.4 mm (Mean± SE= 50.84±4.61 mm) in horses and 37.9 to 51.6 mm (Mean± SE = 42.76±2.34 mm) in mules. Therefore detailed ultrasonographic examination of liver and spleen helped us to compile baseline data, which will be helpful in management of the affections of liver and spleen in future. [ABSTRACT FROM AUTHOR]

The objective of this study was to develop baseline topographical data of landmarks for locating and generating sonographic calliper measurements along with dynamic parameters of caecum and colon in 50 native adult healthy unsedated Spiti horses and Himalayan Hill mules of either sex. The wall thickness, contractility, character and echo-architecture of caecum and colon were recorded. Caecum was found from 15th to 17th inter costal space (ICS). Right dorsal colon (RDC), right ventral colon (RVC), left ventral colon (LVC) and left dorsal colon (LDC) were found from 6th to 14th, 9th to 17th, 9th to 14th and 6th to 15th ICS, respectively. Descending colon was found in the left paralumbar fossa upto tuber-coxae in both horses as well as mules. The corresponding values of horses and mules for wall thickness (in mm) was 2.1±0.05 and 1.4±0.05 for the caecum, 2.16±0.05 and 1.32±0.02 for the right dorsal colon, 2.1±0.03 and 1.32 ±0.02 for the right ventral colon, 2.12±0.03 and 1.32±0.02 for the left dorsal colon, 2.1±0.03 and 1.32±0.02 for the right dorsal colon and 2.04±0.02 for the descending colon. Prominent sacculations were observed on ventral colon and absent on dorsal colon. Detailed ultrasonographic examination of caecum and colon generated baseline data, which will be useful for management of the affections of caecum and colon. [ABSTRACT FROM AUTHOR]

The study was conducted on equids of high altitude to develop baseline topographical data of landmarks, sonographic calliper measurements and dynamic parameters of abdominal organs. Ultrasonography was performed on healthy adult spiti horses and Himalayan mules of either sex. The wall thickness, contractility, character and echo-architecture of these organs were recorded for standardization. The stomach was found between 8th-15th and 8th-14th Inter-Costal Space (ICS) with wall thickness of 4.92 ± 0.23 and 4.24 ± 0.06 mm in horses and mules, respectively. Ascending duodenum was found between 11th-13th and 10th-13th ICS with a wall thickness of 2.0 ± 0.00 and 1.96 ± 0.04 mm in horses and mules, respectively. Descending duodenum was found from 16th and 15th ICS to just caudal to the last rib in horses and mules, respectively. Jejunum was found from 12th (±1) to 17th ICS with wall thickness of 2.04 ± 0.04 and 2.02 ± 0.02 mm in horses and mules, respectively. Detailed ultrasonographic examination of these organs helped to compile baseline data, which will be useful in future in management of the affections of stomach, duodenum and jejunum. [ABSTRACT FROM AUTHOR]

Protamines (PRMs), important proteins of chromatin condensation in spermiogenesis, are promising candidate genes to explore markers of sperm motility. The coding and in-silico predicted promoter regions of these genes were investigated in 102 crossbred and 32 purebred cattle. Also, mRNA quantification was done to explore its possibility as diagnostic tool of infertility. The PCR-SSCP analysis indicated there were two band patterns only in fragment I of the PRM1 and fragment II of the PRM2 gene. The sequence analysis revealed A152G and G179A transitions in the PRM1 gene. Similarly, G35A, A49G and A64G transitions were identified in the PRM2 gene which resulted in altered amino acid sequences from arginine (R) to glutamine (Q), from arginine (R) to glycine (G) and from arginine (R) to glycine (G), respectively. This caused the reduction in molecular weight of PRM2 from 2157.66 to 1931.33 Da due to reduction in the number of basic amino acids. These altered properties of the PRM2 protein led to the reduction in Mass Motility (MM: P  < 0.01), Initial Progressive Motility (IPM; P  < 0.05) and Post Thaw Motility (PTM; P  < 0.05) in crossbred bulls. The least squares analysis of variance indicated there was an effect of PRM2 haplotypes on MM ( P  = 0.0069), IPM ( P  = 0.0306) and PTM ( P  = 0.0500) in crossbred cattle and on PTM ( P  = 0.0408) in the overall cattle population. Based on the RT-qPCR analysis, however, there was not any significant variation of PRM1 and PRM2 gene expression among sperm of Vrindavani bulls with relatively lesser and greater sperm motility. [ABSTRACT FROM AUTHOR]

Vector-borne haemoparasitic diseases of migratory small ruminants have been less explored to date. Therefore, transhumant migratory sheep and goats of the Himalayas were molecularly screened for various vector-borne pathogens like Anaplasma, Theileria , Babesia , Anaplasma marginale , Theileria luwenshuni , Theileria orientalis, and Trypanosoma evansi using their specific primers. Out of the 171 animals screened, 73.09% of the animals were infected with Anaplasma spp. and 74.85% of the animals with Theileria spp., while no animals were found infected with Babesia spp. and Trypanosoma evansi. In a quest to identify the species, a few positive PCR amplicons representing Anaplasma spp., Theileria spp., and Theileria/Babesia spp. were sequenced. The sequences obtained were further BLAST analyzed to reveal maximum identity with Anaplasma capra (100%), Theileria luwenshuni (99.71%), and Theileria sinensis (98.73%). Among them Theileria luwenshuni was further confirmed with their species-specific PCR and the positive amplicons were sequenced and BLAST analysed to reveal their maximumsimilarity with Chinese isolates rather than Indian isolates. Further, phylogenetic analyses of the resulting sequences were conducted to identify their evolutionary history. It revealed that 1100 bp- amplicons of Theileria spp. were showing maximum likeness to Theileria sp. MK, a non-transforming Theileria. This is the first report of zoonotic Anaplasma capra and Theileria sp. MK in small ruminants from India and it could be mostly due to their transboundary migration history through the Indo-Tibetan border during summer seasons as a part of the traditional migratory system of livestock rearing practice in the Himalayas. The high prevalence of haemoparasites in migratory small ruminants calls for the adoption of effective control measures. • First report of Anaplasma capra , Theileria sp. MK from migratory small ruminants of India. • Anaplasma capra is zoonotic and therefore demands utmost attention. • Theileria sp. MK is a non-transforming Theileria closely related to Theileria orientalis. [ABSTRACT FROM AUTHOR]

The present study was conducted to identify the differentially expressed miRNAs (DE miRNAs) in the peripheral blood mononuclear cells of crossbred pigs in response to CSF vaccination on 7 and 21 days of post vaccination as compared to unvaccinated control (0 dpv). Simultaneously, set of miRNA was predicted using mRNA seq data at same time point. The proportion of CD4−CD8+ and CD4+CD8+ increased after vaccination, and the mean percentage inhibition was 86.89% at 21 dpv. It was observed that 22 miRNAs were commonly expressed on both the time points. Out of predicted DE miRNAs, it was found that 40 and 35 DE miRNAs were common, obtained from miRNA seq analysis and predicted using mRNA seq data on 7 dpv versus 0 dpv and 21 dpv versus 0 dpv respectively. Two DE miRNAs, ssc-miR-22-5p and ssc-miR-27b-5p, were selected based on their log2 fold change and functions of their target genes in immune process/pathway of viral infections. The validations of DE miRNAs using qRT-PCR were in concordance with miRNA seq analysis. Two set of target genes, CD40 and SWAP70 (target gene of ssc-miR-22-5p) and TLR4 and Lyn (target gene of ssc-miR-27b-5p), were validated and were in concordance with results of RNA seq analysis at a particular time point (except TLR4). The first report of genome-wide identification of differentially expressed miRNA in response to live attenuated vaccine virus of classical swine fever revealed miR-22-5p and miR-27b-5p were differentially expressed at 7 dpv and 21 dpv. [ABSTRACT FROM AUTHOR]

Since carotenoids are important as natural colorants, antioxidants, neutraceutics and pharmaceutics, the aim of the present study was to find a new good source of these pigments. We hereby report a green microalga Asterarcys quadricellulare PUMCC 5.1.1 as a new and good producer of carotenoids. The organism produced 35±1.75 μg carotenoids mg−1 dry biomass during stationary phase in control cultures. The growth and carotenoids production by the test microalga were optimized by varying nutrient growth media, pH, nitrogen and phosphate source, salinity, light quality, intensity and duration. The optimized conditions for carotenoid production were: Bold basal (BB) medium with pH 8.5, containing with10 mM nitrate, 3.5 mM phosphate and 0.17 mM salinity and illuminated with blue light with 60 μmol m-2 s-1 photon flux light intensity. Cultivation of cultures in the above mentioned optimized conditions resulted in nearly 3.0 fold increase in carotenoid production compared to the control cultures grown in unmodified BB medium. Using HPTLC, four carotenoids have been identified as β-carotene, lutein, astaxanthin and canthaxanthin. Further, carotenoids were also separated and purified by flash chromatography and the amounts of purified carotenoids were determined by HPLC. The organism produced 47.0, 28.7, 15.5 and 14.0 μg β-carotene, lutein, astaxanthin and canthaxanthin mg−1 dry biomass, respectively, under optimized conditions. The amount of total carotenoids (118 μg mg-1 dry biomass) produced by Asterarcys quadricellulare PUMCC 5.1.1 under optimized culture conditions was significantly higher than control cultures. Thus, this microalgal strain is a promising candidate for carotenoid production at commercial level. [ABSTRACT FROM AUTHOR]

The objective of this study was to develop baseline topographical data of landmarks for locating kidneys and urinary bladder and to generate sonographic calliper measurements along with dynamic parameters of these organs. Ultrasonography was performed on 50 native adult (Spiti horses and Himalayan hill mules) healthy unsedated equines of either sex. The thickness, character and echo-architecture of these organs were recorded. The right kidney was found from 15th (±1) intercostal space (ICS) extending upto first lumbar vertebra, both in horses as well as in mules. The height of right kidney varied from 112.7 to 118.3 mm (Mean± SE= 115.6±1.01 mm) and its thickness varied from 58.4 to 71.2 mm (Mean± SE= 64.3±2.37 mm). While in mules height of right kidney varied from 90.7 to 109.2 mm (Mean± SE= of 101.1±3.14 mm) and its thickness varied from 47.3 to 56.8 mm (Mean± SE= 52.3±1.57 mm). The left kidney in horses was found between 16th ICS to 2nd lumbar vertebra, while in mules it was found from 16th ICS to 3rd lumbar vertebra. The height of left kidney varied from 87 to 107.8 mm (Mean± SE= 99.0±3.67 mm) and its thickness varied from 49.5 to 53.8 mm (Mean± SE= 51.8±0.72 mm). In mules the height of left kidney varied from 85.5 to 111.4 mm (Mean± SE= 95.3±2.36 mm) and its thickness varied from 43.4 to 55.7 mm (Mean± SE= 50.6±2.53 mm). During transrectal ultrasonography urinary bladder was observed as an oval to round structure in the caudal ventral abdomen, the wall thickness of urinary bladder varied from 2.7 to 2.8 mm (Mean± SE= 2.74±0.02 mm), 2.4 to 2.7 mm (Mean± SE= 2.62±0.05 mm) in horses and mules respectively. Hence detailed ultrasonographic examination of these organs helped us to compile baseline data, which will be helpful in future in management of the affections of these organs. [ABSTRACT FROM AUTHOR]

[Display omitted] • Adsorption energy, UV–Visible spectrum, Reduced Density Gradient (RDG), HOMO-LUMO gap were calculated for all systems. • The Au7Cu nanoclusters and their complexes with glycine have maximum HOMO-LUMO gap therefore they have more stability or less reactivity in comparison to the other nanoclusters in this work. • The maximum adsorption energy is found in the small copper doped gold nanoclusters [Au 4 Cug(A,B)] and in spherically shaped thirteen atoms nanoclusters [Au 12 Cug(C)]. • Five and fifteen-atom nanoclusters could be observed red shift and eight, thirteen, and eighteen atoms and their complexes have shown a blue shift in the UV–Visible spectra when glycine is adsorbed on Cu doped Au nanoclusters. Gold nanoclusters and bimetallic gold nanoclusters are interesting areas of research. These nanoclusters are used for the detection of organic molecules, cancer treatment, bio-imaging, drug delivery etc. In this paper, we studied the interaction of copper doped Au nanoclusters of five, eight, thirteen, fifteen, and eighteen atoms with glycine amino acid. Their HOMO-LUMO gap, adsorption energy, non-covalent interaction, hardness, electrophilicity index, electrochemical potential, and UV–Visible spectra were analyzed. The maximum adsorption energy is found in the nanoclusters Au 4 Cug(A,B) and Au 12 Cug(C). Additionally, strong non-covalent interaction is observed in the Au 4 Cug(A,B) and Au 12 Cug(B) nanoclusters, which makes them suitable for applications like drug delivery, biosensing etc. These nanoclusters are also more stable compared to other nanoclusters in this work (only exception Au 7 Cu(A,B,C)) owing to their large HOMO-LUMO gap and symmetric structure. [ABSTRACT FROM AUTHOR]

The present study in the 5’ upstream region of TLR4 gene revealed four Single Nucleotide Polymorphisms (SNPs) in Vrindavani and Tharparkar cattle. The polymorphic information content (PIC), heterozygosity and allelic diversity values were low to moderate for these SNPs. In Vrindavani cattle, one SNP was found to be in Hardy-Weinberg Equilibrium (HWE) and the remaining three were found to be in linkage disequilibrium (LD) as indicated statistically (P > 0.05). In Tharparkar cattle, two SNPs were found to be in HWE and were not in LD as indicated statistically (P > 0.05). These SNPs were used for construction of haplotypes. In-silico analysis of these SNPs predicted abolition of eight transcription factor binding sites and creation of eight new sites. The quantitative real time PCR analysis did not show any significant variation of gene expression among haplotypes. However, gene expression between breed was found to be significant (P < 0.05) which suggested that upstream region of bovine TLR4 gene has a crucial role in its expression. These findings in TLR4 gene offer essential evidence that can be useful in future research exploring its role in immunity. TLR4 can be used as a marker for selection for disease resistance in bovines. [ABSTRACT FROM AUTHOR]

122 randomly selected Vrindavani cattle were studied to detect polymorphism in four fragments of theCatSper2gene that were comprised of exon 2, 4, 5, and 6 with flanking regions. Using PCR-SSCP and sequencing analysis, three SNPs (T157C, C273A, and A274C) in the first fragment, one SNP (C30G) in the second fragment, and two SNPs (T86G and T292C) in the fourth fragment were identified. The third fragment did not reveal any polymorphism. The SNPs were used for construction of haplotypes and three haplotypes were found. The least square analysis of variance revealed a significant (P < 0.01) effect of haplotype on all three motility parameters. The haplotype II and III were nonsignificantly different from each other while being significantly (P < 0.01) different from haplotype I. The nonsignificant difference of haplotype II with III can lead to a hypothesis that T>G or C>T SNPs may not play a role in sperm motility. However, when the comparison was made between haplotype I and II, it can be inferred that C>T SNP may have a role in sperm motility, as haplotype II has better motility parameters. Expression profiling ofCatper2gene revealed nonsignificant down regulation ofCatSper2gene in poor motility sperm compared to good motility sperm. [ABSTRACT FROM AUTHOR]

Crossbred cattle in some sectors of the world have a significant role in enhancing milk production thereby enhancing the per capita milk availability as a human food source. However, there are certain constraints associated with crossbred animals, such as disease susceptibility, increased reproductive problems, repeat breeding and poor seminal quality. The semen of crossbred bulls has a poor freezing capacity, increased cryo-damage, poor mass cell motility, greater percentages of dead/abnormal sperm and poor initial and post-freeze cell motility. The rejection rate of crossbred bulls for cryostorage of semen has been reported to be as great as 50% as a result of unacceptable semen quality. The identification of superior bulls using molecular technologies is needed which necessitates identification of the genes having a role in sperm function. The present study was, therefore, conducted to gain information on identification and expression of genes having a role in sperm motility in crossbred bulls. The gene transcripts in bulls with sperm of superior and inferior quality were profiled in Vrindavani crossbred cattle by microarray analyses and the results were verified by real time-quantitative PCR. Microarray analyses revealed 19,454 genes which were differentially expressed. At a two-fold cut off, 305 genes were differentially ( P < 0.01) expressed with 160 genes upregulated and 145 genes down regulated. Some of the upregulated candidate genes were further validated by RT-qPCR. These genes had a four to 16 fold upregulation in sperm with inferior motility as compared to sperm of crossbred bulls with superior motility. [ABSTRACT FROM AUTHOR]

The exploration of candidate genes for immune response in cattle may be vital for improving our understanding regarding the species specific response to pathogens. Toll-like receptor 4 (TLR4) is mostly involved in protection against the deleterious effects of Gram negative pathogens. Approximately 2.6 kb long cDNA sequence of TLR4 gene covering the entire coding region was characterized in two Indian milk cattle (Vrindavani and Tharparkar). The phylogenetic analysis confirmed that the bovine TLR4 was apparently evolved from an ancestral form that predated the appearance of vertebrates, and it is grouped with buffalo, yak, and mithun TLR4s. Sequence analysis revealed a 2526-nucleotide long open reading frame (ORF) encoding 841 amino acids, similar to other cattle breeds. The calculated molecular weight of the translated ORF was 96144 and 96040.9 Da; the isoelectric point was 6.35 and 6.42 in Vrindavani and Tharparkar cattle, respectively. The Simple Modular Architecture Research Tool (SMART) analysis identified 14 leucine rich repeats (LRR) motifs in bovine TLR4 protein. The deduced TLR4 amino acid sequence of Tharparkar had 4 different substitutions as compared toBos taurus, Sahiwal, and Vrindavani. The signal peptide cleavage site predicted to lie between 16th and 17th amino acid of mature peptide. The transmebrane helix was identified between 635–657 amino acids in the mature peptide. [ABSTRACT FROM AUTHOR]