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We reported previously that the methanolic root extract of the Indian medicinal plant Pluchea indica Less. (Asteraceae) could neutralize viper venom-induced action [Alam, M.I., Auddy, B., Gomes, A., 1996. Viper venom neutralization by Indian medicinal plant (Hemidesmus indicus and P. indica) root extracts. Phytother. Res. 10, 58-61]. The present study reports the neutralization of viper and cobra venom by beta-sitosterol and stigmasterol isolated from the root extract of P. indica Less. (Asteraceae). The active fraction (containing the major compound beta-sitosterol and the minor compound stigmasterol) was isolated and purified by silica gel column chromatography and the structure was determined using spectroscopic analysis (EIMS, (1)H NMR, (13)C NMR). Anti-snake venom activity was studied in experimental animals. The active fraction was found to significantly neutralize viper venom-induced lethal, hemorrhagic, defibrinogenation, edema and PLA(2) activity. Cobra venom-induced lethality, cardiotoxicity, neurotoxicity, respiratory changes and PLA(2) activity were also antagonized by the active component. It potentiated commercial snake venom antiserum action against venom-induced lethality in male albino mice. The active fraction could antagonize venom-induced changes in lipid peroxidation and superoxide dismutase activity. This study suggests that beta-sitosterol and stigmasterol may play an important role, along with antiserum, in neutralizing snake venom-induced actions. [ABSTRACT FROM AUTHOR]

Precise as well as unbiased estimates of genetic parameters of economically important traits is important for framing breeding programmes meant for genetic progress of dairy animals. Present work was carried out to assess the co-variances along with genetic parameters of production and reproduction traits in Sahiwal cattle with Bayesian approach. Records of Sahiwal cattle kept at ICAR-National Dairy Research Institute, Karnal, Haryana for the period 1988-2016 were used. Heritability estimates ± posterior standard deviation for 305 days' milk yield (305DMY), lactation milk yield (LMY), lactation length (LL), peak yield (PY), days to attain peak yield (DPY), dry period (DP), gestation period (GP), calving to first insemination interval (CFI), days open (DO) and calving interval (CI) as obtained were 0.23 ± 0.03, 0.26 ± 0.03, 0.18 ± 0.03, 0.18 ± 0.03, 0.07 ± 0.02, 0.08 ± 0.02, 0.08 ± 0.02, 0.07 ± 0.02, 0.09 ± 0.02 and 0.02 ± 0.02, respectively and estimates of repeatability for the corresponding traits were 0.23 ± 0.03, 0.26 ± 0.03, 0.21 ± 0.03, 0.21 ± 0.03, 0.12 ± 0.02, 0.14 ± 0.03, 0.16 ± 0.03, 0.16 ± 0.03, 0.17 ± 0.03 and 0.17 ± 0.03. Moderate heritability values for 305DMY and LMY implies that sufficient additive genetic variability is available to obtain selection response. Repeatability estimates for 305DMY, LMY and PY were moderate, pointing towards the possibility of early appraisal of Sahiwal cattle for selection. Antagonistic genetic associations were observed between reproduction and production traits suggesting that reproduction traits need due attention while selecting high producing Sahiwal animals., (© 2024. The Author(s), under exclusive licence to Springer Nature B.V.)

Present study was conducted to investigate the impact of temperature humidity index (THI) on expected breeding value (EBV) for daily milk yield (DMY) and monthly test day fat% (MTDF%) and monthly test day SNF% (MTDSNF%) of Murrah buffaloes at ICAR-National Dairy Research Institute, Karnal. 302,101 records of DMY and 9864 records each on MTDF% and MTDSNF% buffaloes spanned over 20 years (1994 to 2013) from NDRI, Karnal, and meteorological were collected from ICAR-CSSRI, Karnal. The year was classified into three zones: non-heat stress (NHSZ), heat stress (HSZ), and critical heat stress zone (CHSZ) based on THI. The heritability (h 2 ) estimates for DMY, MTDF%, and MTDSNF% in NHSZ were 0.432 ± 0.054, 0.090 ± 0.004, and 0.070 ± 0.002; in HSZ 0.491 ± 0.073, 0.112 ± 0.003, and 0.052 ± 0.001; and in CHSZ 0.524 ± 0.077, 0.116 ± 0.004, and 0.092 ± 0.003, respectively. The repeatability (r) for DMY, MTDF%, and MTDSNF% in NHSZ were 0.528 ± 0.006, 0.166 ± 0.007, and 0.135 ± 0.007; in HSZ 0.572 ± 0.007, 0.198 ± 0.006, and 0.077 ± 0.006; and in CHSZ 0.599 ± 0.008, 0.217 ± 0.004, and 0.156 ± 0.009, respectively. EBV for DMY and MTDF% was maximum in NHSZ (8.85 kg and 7.85%) and in HSZ (7.27 kg and7.78%) and lowest (6.90 kg and 7.77%) at CHSZ. For MTDSNF%, EBV was highest during NHSZ (9.6403), declined to 9.6265 at HSZ, and marginally elevated to 9.6271 at CHSZ. Impact of climatic factors on milk production and constituent traits is vital, and proper management should be followed during the heat stress and critical heat stress periods to improve the production performance of Murrah buffaloes., (© 2023. The Author(s), under exclusive licence to Springer Nature B.V.)

Multicellularity was a watershed development in evolution. However, it also meant that individual cells could escape regulatory mechanisms that restrict proliferation at a severe cost to the organism: cancer. From the standpoint of cellular organization, evolutionary complexity scales to organize different molecules within the intracellular milieu. The recent realization that many biomolecules can "phase-separate" into membraneless organelles, reorganizing cellular biochemistry in space and time, has led to an explosion of research activity in this area. In this review, we explore mechanistic connections between phase separation and cancer-associated processes and emerging examples of how these become deranged in malignancy., Significance: One of the fundamental functions of phase separation is to rapidly and dynamically respond to environmental perturbations. Importantly, these changes often lead to alterations in cancer-relevant pathways and processes. This review covers recent advances in the field, including emerging principles and mechanisms of phase separation in cancer., (©2022 The Authors; Published by the American Association for Cancer Research.)

An attempt was made for the first time in India to develop the performance indices for quantifying the relative emphasis to be given to production, reproduction, health and longevity traits for selection and genetic improvement of Sahiwal cattle in a sustainable manner. The study was conducted using information related to various production, reproduction, health and longevity traits spread over a period of 29 years in Sahiwal cattle. Using income and expenditure method, the relative economic values for 305 days milk yield (305DMY), average daily milk yield (ADMY), calving to first insemination interval (CFI), days open (DO) and longevity (LNG) were estimated as 1, 1, -6.62, -6.62 and 5.96 in Sahiwal cattle. A total of three performance indices were constructed using three-trait combination, that is production, reproduction and longevity before and after excluding the days suffered by the animals. Correlation between the aggregate genotype and index (R IH ) was computed to determine the accuracy of each performance index, and based on highest R IH value, best performance index was identified for selection of Sahiwal cattle. Developed indices were further assessed for robustness by increasing the relative economic values of the traits by 25% and 50%. The performance index (305DMY, DO and LNG) was found to be the best index before as well as after excluding days suffered by the animals, but in later case emphasis to be given to different traits was found to be more balanced. The best index indicated that about 47, 42 and 11% relative emphasis to be assigned to 305 days milk yield, days open and longevity for selection of Sahiwal animals for sustainable genetic improvement., (© 2021 Wiley-VCH GmbH.)

Bull fertility is considered an indispensable trait, as far as farm economics is concerned since it is the successful conception in a cow that provides calf crop, along with the ensuing lactation. This ensures sustainability of a dairy farm. Traditionally, bull fertility did not receive much attention by the farm managers and breeding animals were solely evaluated based on phenotypic predictors, namely, sire conception rate and seminal parameters in bull. With the advent of the molecular era in animal breeding, attempts were made to unravel the genetic complexity of bull fertility by the identification of genetic markers related to the trait. Marker-Assisted Selection (MAS) is a methodology that aims at utilizing the genetic information at markers and selecting improved populations for important traits. Traditionally, MAS was pursued using a candidate gene approach for identifying markers related to genes that are already known to have a physiological function related to the trait but this approach had certain shortcomings like stringent criteria for significance testing. Now, with the availability of genome-wide data, the number of markers identified and variance explained in relation to bull fertility has gone up. So, this presents a unique opportunity to revisit MAS by selection based on the information of a large number of genome-wide markers and thus, improving the accuracy of selection.

The GH has a multifaceted role in growth, reproduction, and milk production. Nonsynonymous mutations identified as L153V were observed from GH1 (L) to GH2 (V) with higher genotypic frequency for GH1 being 0.87. GH2 (V) allele was identified as a rare allele and the population followed Hardy-Weinberg equilibrium. LL homozygote variant had significantly better growth, reproduction, and expected milk production at different ages in crossbred (CB) males. Reports are scanty explaining the molecular mechanism of how individuals with LV genotype were phenotypically inferior to that of wild-type LL. In the present study, it was explored that GH peptide with LV heterozygotes of GH gene, were observed to have reduced structural stability thermodynamically and thus functionally leads to reduced economic traits in CB animals. The fact was first time reported and confirmed through genomic analysis, bioinformatics, and later confirmed through immunohistochemistry. Differential expression analysis of the GH gene with respect to other genes in the hypothalamus-pituitary growth axis of CB cattle was also studied to have a complete insight of the GH gene., (Copyright © 2019 Elsevier Inc. All rights reserved.)

Covalent modifications to histones are essential for development, establishing distinct and functional chromatin domains from a common genetic sequence. Whereas repressed chromatin is robustly inherited, no mechanism that facilitates inheritance of an activated domain has been described. Here, we report that the Set3C histone deacetylase scaffold Snt1 can act as a prion that drives the emergence and transgenerational inheritance of an activated chromatin state. This prion, which we term [ESI + ] for expressed sub-telomeric information, is triggered by transient Snt1 phosphorylation upon cell cycle arrest. Once engaged, the prion reshapes the activity of Snt1 and the Set3C complex, recruiting RNA pol II and interfering with Rap1 binding to activate genes in otherwise repressed sub-telomeric domains. This transcriptional state confers broad resistance to environmental stress, including antifungal drugs. Altogether, our results establish a robust means by which a prion can facilitate inheritance of an activated chromatin state to provide adaptive benefit., Competing Interests: Declaration of Interests The authors declare no competing interests., (Copyright © 2020 Elsevier Inc. All rights reserved.)

Theory and experiments suggest that organisms would benefit from pre-adaptation to future stressors based on reproducible environmental fluctuations experienced by their ancestors, but the mechanisms driving pre-adaptation remain enigmatic. We report that the [SMAUG + ] prion allows yeast to anticipate nutrient repletion after periods of starvation, providing a strong selective advantage. By transforming the landscape of post-transcriptional gene expression, [SMAUG + ] regulates the decision between two broad growth and survival strategies: mitotic proliferation or meiotic differentiation into a stress-resistant state. [SMAUG + ] is common in laboratory yeast strains, where standard propagation practice produces regular cycles of nutrient scarcity followed by repletion. Distinct [SMAUG + ] variants are also widespread in wild yeast isolates from multiple niches, establishing that prion polymorphs can be utilized in natural populations. Our data provide a striking example of how protein-based epigenetic switches, hidden in plain sight, can establish a transgenerational memory that integrates adaptive prediction into developmental decisions., (Copyright © 2019 Elsevier Inc. All rights reserved.)

Spatiotemporal gene regulation is often driven by RNA-binding proteins that harbor long intrinsically disordered regions in addition to folded RNA-binding domains. We report that the disordered region of the evolutionarily ancient developmental regulator Vts1/Smaug drives self-assembly into gel-like condensates. These proteinaceous particles are not composed of amyloid, yet they are infectious, allowing them to act as a protein-based epigenetic element: a prion [SMAUG + ]. In contrast to many amyloid prions, condensation of Vts1 enhances its function in mRNA decay, and its self-assembly properties are conserved over large evolutionary distances. Yeast cells harboring [SMAUG + ] downregulate a coherent network of mRNAs and exhibit improved growth under nutrient limitation. Vts1 condensates formed from purified protein can transform naive cells to acquire [SMAUG + ]. Our data establish that non-amyloid self-assembly of RNA-binding proteins can drive a form of epigenetics beyond the chromosome, instilling adaptive gene expression programs that are heritable over long biological timescales., (Copyright © 2019 Elsevier Inc. All rights reserved.)

Background: The bovine AGPAT6 gene is one of the potential candidate genes governing milk fat synthesis., Objectives: Identification of single nucleotide polymorphisms (SNP) in the targeted region of AGPAT6 gene and their effect on expected breeding values (EBV) of first lactation milk production traits viz. fat %, fat yield and 305 days milk yield in Karan Fries (KF) breeding bulls were sought., Materials and Methods: A tetra-primer ARMS PCR technique was adapted to genotype an SNP, g.36,175,805C>T located on 5' flanking region of AGPAT6 gene. The relationship between EBV of milk production traits and polymorphic locus of AGPAT6 gene was assessed., Results: Three kinds of genotype (CC, CT, and TT) with respect to g.36,175,805C>T SNP locus were observed. The identified SNP had significant (P<0.05) influence on EBV of fat % (EBV-FP). The KF bulls with CC and CT genotype had comparatively higher EBV-FP than the bulls with the TT genotype. The substitution of "C" allele by "T" allele led to a decrease of 0.0045 % in the EBV-FP., Conclusion: The identified SNP was significantly associated with EBV-FP, thus it may be utilized as a molecular marker for developing marker-assisted selection strategy to enhance the milk fat content in KF cattle population., (Copyright: © 2019 The Author(s); Published by Iranian Journal of Biotechnology.)

A central tenet of molecular biology is that heritable information is stored in nucleic acids. However, this paradigm has been overturned by a group of proteins called "prions." Prion proteins, many of which are intrinsically disordered, can adopt multiple conformations, at least one of which has the capacity to self-template. This unusual folding landscape drives a form of extreme epigenetic inheritance that can be stable through both mitotic and meiotic cell divisions. Although the first prion discovered-mammalian PrP-is the causative agent of debilitating neuropathies, many additional prions have now been identified that are not obviously detrimental and can even be adaptive. Intrinsically disordered regions, which endow proteins with the bulk property of "phase-separation," can also be drivers of prion formation. Indeed, many protein domains that promote phase separation have been described as prion-like. In this review, we describe how prions lie at the crossroads of phase separation, epigenetic inheritance, and evolutionary adaptation., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

The thermodynamic stability of biomolecules in the perspective of evolution is a complex issue and needs discussion. Intra molecular bonds maintain the structure and the state of internal energy (E) of a biomolecule at "local minima". In this communication, possibility of loss in internal energy level of a biomolecule through the changes in the bonds has been discussed, that might earn more thermodynamic stability for the molecule. In the process variations in structure and functions of the molecule could occur. Thus, E of a biomolecule is likely to have energy stature for minimization. Such change in energy status is an intrinsic factor for evolving biomolecules buying more stability and generating variations in the structure and function of DNA molecules undergoing natural selection. Thus, the variations might very well contribute towards the process of evolution. A brief discussion on conserved sequence in the light of proposition in this communication has been made at the end. Extension of the idea may resolve certain standing problems in evolution, such as maintenance of conserved sequences in genome of diverse species, pre- versus post adaptive mutations, 'orthogenesis', etc., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

RNA-binding proteins (RBPs) control the fate of nearly every transcript in a cell. However, no existing approach for studying these posttranscriptional gene regulators combines transcriptome-wide throughput and biophysical precision. Here, we describe an assay that accomplishes this. Using commonly available hardware, we built a customizable, open-source platform that leverages the inherent throughput of Illumina technology for direct biophysical measurements. We used the platform to quantitatively measure the binding affinity of the prototypical RBP Vts1 for every transcript in the Saccharomyces cerevisiae genome. The scale and precision of these measurements revealed many previously unknown features of this well-studied RBP. Our transcribed genome array (TGA) assayed both rare and abundant transcripts with equivalent proficiency, revealing hundreds of low-abundance targets missed by previous approaches. These targets regulated diverse biological processes including nutrient sensing and the DNA damage response, and implicated Vts1 in de novo gene "birth." TGA provided single-nucleotide resolution for each binding site and delineated a highly specific sequence and structure motif for Vts1 binding. Changes in transcript levels in vts1 Δ cells established the regulatory function of these binding sites. The impact of Vts1 on transcript abundance was largely independent of where it bound within an mRNA, challenging prevailing assumptions about how this RBP drives RNA degradation. TGA thus enables a quantitative description of the relationship between variant RNA structures, affinity, and in vivo phenotype on a transcriptome-wide scale. We anticipate that TGA will provide similarly comprehensive and quantitative insights into the function of virtually any RBP.

Intense selection of buffaloes for milk production at organized herds of the country without giving due attention to fertility traits has lead to deterioration in their performances. The study was initiated to assess the relationship between milk production and productivity with fertility in Murrah buffaloes. In the present study, fertility was defined in terms of pregnancy rate of Murrah buffaloes. Pregnancy rate measures the percentage of non-pregnant animals that become pregnant during each oestrous cycle. Data pertaining to 1224 lactation records of Murrah buffaloes spread over a period 22 years from 1993 to 2014 were analyzed in the study. It was observed that pregnancy rate is negatively associated with 305days or less milk yield (-0.08±0.04) and wet average (-0.12±0.02) and positively associated with life time (0.15±0.03) in Murrah buffaloes. Wet average is defined as average daily milk yield per lactation of Murrah buffaloes. To achieve around 2000kg 305days or less milk yield and 7.5kg wet average, the level of pregnancy rate in Murrah buffaloes should vary between 30 and 50%. The per unit change in pregnancy rate with respect to milk yield in Murrah buffaloes, were studied using regression analysis. The results suggest that increasing hundred kilogram in 305days or one kilogram milk yield per day in 305days, pregnancy rate reduced by 0.9% in overall lactations of Murrah buffaloes. By increasing hundred kilogram life time 305days or less milk yield and one kilogram life time wet average, pregnancy rate of Murrah buffaloes reduced by about 0.2% and 0.34%, respectively. The study quantifies the decline of pregnancy rate with increase of lactation milk yield and wet average in Murrah buffaloes and emphasis the importance of fertility i.e. pregnancy rate in the evaluation and breeding programmes of Murrah buffaloes., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Aim: The present study was performed to investigate the effect of genetic and non-genetic factors affecting milk fat yield and to estimate genetic parameters of monthly test day fat yields (MTDFY) and lactation 305-day fat yield (L305FY) in Murrah buffaloes., Materials and Methods: The data on total of 10381 MTDFY records comprising the first four lactations of 470 Murrah buffaloes calved from 1993 to 2014 were assessed. These buffaloes were sired by 75 bulls maintained in an organized farm at ICAR-National Dairy Research Institute, Karnal. Least squares maximum likelihood program was used to estimate genetic and non-genetic parameters. Heritability estimates were obtained using paternal half-sib correlation method. Genetic and phenotypic correlations among MTDFY, and 305-day fat yield were calculated from the analysis of variance and covariance matrix among sire groups., Results: The overall least squares mean of L305FY was found to be 175.74±4.12 kg. The least squares mean of overall MTDFY ranged from 3.33±0.14 kg (TD-11) to 7.06±0.17 kg (TD-3). The h(2) estimate of L305FY was found to be 0.33±0.16 in this study. The estimates of phenotypic and genetic correlations between 305-day fat yield and different MTDFY ranged from 0.32 to 0.48 and 0.51 to 0.99, respectively., Conclusions: In this study, all the genetic and non-genetic factors except age at the first calving group, significantly affected the traits under study. The estimates of phenotypic and genetic correlations of MTDFY with 305-day fat yield was generally higher in the MTDFY-5 of lactation suggesting that this TD yields could be used as the selection criteria for early evaluation and selection of Murrah buffaloes.

Heat stress has adverse effects on the reproductive performances of dairy cattle and buffaloes. The dairy sector is a more vulnerable to global warming and climate change. The temperature humidity index (THI) is the widely used index to measure the magnitude of heat stress in animals. The objective of this paper was to assess the decline in performances of reproductive traits such as service period, conception rate and pregnancy rate of dairy cattle and buffaloes with respect to increase in THI. The review stated that service period in cattle is affected by season of calving for which cows calved in summer had the longest service period. The conception rate and pregnancy rate in dairy cattle were found decreased above THI 72 while a significant decline in reproductive performances of buffaloes was observed above threshold THI 75. The non-heat stress zone (HSZ) (October to March) is favorable for optimum reproductive performance, while fertility is depressed in HSZ (April to September) and critical HSZ (CHSZ) (May and June). Heat stress in animals has been associated with reduced fertility through its deleterious impact on oocyte maturation and early embryo development. The management strategies viz., nutrition modification, environment modification and timed artificial insemination protocol are to be strictly operated to ameliorate the adverse effects of heat stress in cattle and buffaloes during CHSZ to improve their fertility. The identification of genes associated with heat tolerance, its incorporation into breeding program and the inclusion of THI covariate effects in selection index should be targeted for genetic evaluation of dairy animals in the hot climate.

In artificial insemination, poor quality of semen unsuitable for cryopreservation and susceptibility of spermatozoa to cryodamage in crossbred bulls have been a matter of concern. Present study was designed to identify the testicular cytology indices that might be used to predict the semen quality and cryotolerance of spermatozoa in bulls. Based on the ejaculate rejection rate and sperm cryotolerance, bulls (Holstein Friesian X Tharparkar crossbred) were classified into either good (producing good quality semen with spermatozoa having good cryotolerance; n = 4) or poor (producing poor quality semen with spermatozoa having poor cryotolerance; n = 4). Testicular cytology was studied in all the 8 bulls using fine needle aspiration technique. Testicular cytology of good bulls and poor bulls differed significantly. The proportion of Sertoli cells was significantly higher in good bulls (25.3 ± 1.6) compared to poor bulls (11.0 ± 0.8). The Sertoli cell index was 46.1 ± 5.0 in good bulls while it was only 13.8 ± 1.3 in poor bulls. The cut off values, as determined using Receiver Operating Characteristics analysis, indicate that the bulls having testicular cytogram comprising of < 15.5% Sertoli cells, < 24.3 Sertoli cell index and > 4.0 spermatogenic cells to Sertoli cell ratio might be a poor bull in terms of semen quality and cryotolerance of spermatozoa. The proportion of Sertoli cells in the testicular cytology had positive (P < 0.05) relationship with semen quality and cryotolerance of spermatozoa.

Aim: The objective of the study was to evaluate the effects of non-genetic factors on reproduction traits viz. age at first semen freezing and age at first semen use of breeding bulls in Sahiwal bulls by fitting least-squares analysis., Materials and Methods: The information on reproduction traits of 43 Sahiwal breeding bulls belonging to 8 sets of Sahiwal breeding program at Indian Council of Agricultural Research-National Dairy Research Institute (ICAR-NDRI), Karnal (Haryana), India during 27 years (1987-2013) were analyzed using fixed linear model. The information was collected from AI records, reproduction sheets, and bull AI register maintained at different sections of Institute viz. record room of Dairy Cattle Breeding Division (DCB), Cattle Yard, Artificial Breeding Research Centre, ICAR-NDRI, Karnal., Results: The average age at first semen freezing and age at first semen use of Sahiwal breeding bulls was estimated as 3.17±0.01 years and 5.35±0.01 years, with the coefficient of variation 18.93% and 20%, respectively. The overall least-squares mean for age at first semen freezing and age at first semen use was estimated as 3.14±0.09 years and 5.25±0.02 years, respectively, in Sahiwal breeding bulls. Period of freezing/use had significant effects on reproductive traits (p<0.01). Season had no significant effect on any of the traits considered in this study., Conclusion: It can be concluded that management inputs such as nutrition, breeding, and optimum environment should be taken care of to optimize age at first semen freezing and age at first semen use for better utilization of superior germplasm.

Heat stress has adverse effects on fertility of dairy animals. Decline in fertility is linearly associated with an increase in combination of both temperature and humidity. The purpose of this study was to investigate the relationship between temperature humidity index (THI) and the pregnancy rate of Murrah buffaloes in a subtropical climate. The effects of genetic and non-genetic factors viz., sire, parity, period of calving and age group at first calving were found non-significant on pregnancy rate. The effect of THI was found significant (p<0.001) on pregnancy rate of Murrah buffaloes calved for first time and overall pregnancy rate. The threshold THI affecting the pregnancy rate was identified as THI 75. The months from October to March showed THI<75 and considered as non heat stress zone (NHSZ), while months from April to September were determined as heat stress zone (HSZ) with THI≥75. The lowest overall pregnancy rate (0.25) was obtained in July with THI 80.9, while the highest overall pregnancy rate (0.59) was found in November with THI 66.1. May and June were identified as critical heat stress zone (CHSZ) within the HSZ with maximum decline (-7%) in pregnancy rate with per unit increase in THI. The highest overall pregnancy rate was estimated as 0.45 in NHSZ with THI value 56.7 to 73.2. The pregnancy rate was found to have declined to 0.28 in HSZ with THI 73.5 to 83.7. However, the lowest pregnancy rate was estimated as 0.27 in CHSZ with THI value 80.3 to 81.6.

Aim: The study was conducted at Artificial Breeding Research Centre, NDRI, Karnal, to determine the sexual behavior and its relationship with semen quality parameters in Sahiwal breeding bulls., Materials and Methods: A total of 63 ejaculates were collected from six adult Sahiwal bulls (age ~47 mo and bwt ~466 kg), to study the relationship of sexual behavior and semen quality. The degree of association between different variables was estimated by Pearson's correlation coefficient method., Results: The results depicted that, sexual aggressiveness showed significantly high positive correlation with libido score (LS) and sexual behavior score (SBS). Reaction time (RT) and total time taken in mounts (TTTM) had a significant negative correlation with LS and SBS. Penile erection score and penile protrusion score (PPS) both had a significant positive correlation with ejaculatory thrust score, mating ability score, and SBS. Results of correlation among seminal attributes and with sexual behavior depicted that ejaculate volume had positive significant correlation with initial progressive motility (IPM), sperm concentration (SCON), head abnormality, total abnormality, hypo-osmotic swelling test (HOST), acrosomal integrity (AI) whereas, mass activity had positive significant correlation with IPM, SCON, non-eosinophilic spermatozoa count (NESC), HOST, AI, RT and TTTM and IPM had positive significant correlation with SCON, NESC, HOST, AI, and TTTM, whereas and HOST had positive significant correlation with AI. Among seminal attributes, SCON had a positive significant correlation with PPS where as head abnormalities had a positive significant correlation with RT and TTTM., Conclusion: It can be concluded that the relationship of sexual behavior and semen quality parameters are reflecting that the sexual behavior of individual bulls is important to harvest good quality and quantity of semen as desired type of sexual preparation can be provided.

In this study genome-wide di-methylated H3K4 (H3K4me2) and tri-methylated H3K27 (H3K27me3) modification profiles were analyzed in spermatozoa of buffalo bulls having wide fertility differences. The custom designed 4 × 180 K buffalo (Bubalus bubalis) ChIP-on-chip array was fabricated by employing array-based sequential hybridization using bovine and buffalo genomic DNA for comparative hybridization. The buffalo specific array developed had 177,440 features assembled from Coding sequences, Promoter and CpG regions comprising 2967 unique genes. A total of 84 genes for H3K4me2 and 80 genes for H3K27me3 were found differentially enriched in mature sperm of high and sub-fertile buffalo bulls. Gene Ontology analysis of these genes revealed their association with different cellular functions and biological processes. Genes identified as differentially enriched between high and sub-fertile bulls were found to be involved in the processes of germ cell development, spermatogenesis and embryonic development. This study presents the first genome-wide H3K4me2 and H3K27me3 profiling of buffalo bull sperm. Results provide a list of specific genes which could be made responsible for differential bull fertility., (© 2014 Wiley Periodicals, Inc.)

Aim: The aim of the present investigation was to optimize the age at first use (AAFU) of semen of Murrah breeding bulls, which will help in early selection of bulls under progeny testing program for improving the reproductive performance in the herd., Materials and Methods: The data on AAFU, conception rate based on first A.I. (CRFAI), overall conception rate (OCR), and birth weight (B.WT) of 57 Murrah bulls during 1993-2014 at NDRI center pertaining to 14 sets of Network Project on Buffalo Improvement at ICAR-National Dairy Research Institute, Karnal, Haryana, India were adjusted for significant environmental influences and subsequently analyzed. Simple and multiple regression models were used for prediction of CRFAI and OCR of Murrah breeding bulls. Comparative evaluation of three developed models (I-III) showed that Model III, having AAFU and B.WT, fulfill the accuracy of model as revealed by high coefficient of determination, low mean sum of squares due to error, low conceptual predictive value, and low Bayesian information criterion., Results: The results revealed that the average predicted CRFAI was highest (39.95%) at <3.5 years and lowest (34.87%) at >4.5 years of age at first A.I/use. Similarly, average predicted OCR was highest (41.05%) at <3.5 years and lowest (39.42%) at >4.5 years of age at first A.I/use of Murrah bulls., Conclusion: In organized herd under progeny testing program, Murrah bulls should be used at young age, i.e. prior to 3.5 years, which is expected to result in 5.08% better CRFAI and 1.63% better OCR in comparison to Murrah bulls used after 4.5 years of age.

Aim: The aim of the present study was to assess the influence of temperature and humidity prevalent under subtropical climate on the breeding values for fertility traits viz. service period (SP), pregnancy rate (PR) and conception rate (CR) of Murrah buffaloes in National Dairy Research Institute (NDRI) herd., Materials and Methods: Fertility data on 1379 records of 581 Murrah buffaloes spread over four lactations and climatic parameters viz. dry bulb temperature and relative humidity (RH) spanned over 20 years (1993-2012) were collected from NDRI and Central Soil and Salinity Research Institute, Karnal, India. Monthly average temperature humidity index (THI) values were estimated. Threshold THI value affecting fertility traits was identified by fixed least-squares model analysis. Three zones of non-heat stress, heat stress and critical heat stress zones were developed in a year. The genetic parameters heritability (h(2)) and repeatability (r) of each fertility trait were estimated. Genetic evaluation of Murrah buffaloes was performed in each zone with respect to their expected breeding values (EBV) for fertility traits., Results: Effect of THI was found significant (p<0.001) on all fertility traits with threshold THI value identified as 75. Based on THI values, a year was classified into three zones: Non heat stress zone(THI 56.71-73.21), HSZ (THI 75.39-81.60) and critical HSZ (THI 80.27-81.60). The EBVfor SP, PR, CR were estimated as 138.57 days, 0.362 and 69.02% in non-HSZ while in HSZ EBV were found as 139.62 days, 0.358 and 68.81%, respectively. EBV for SP was increased to 140.92 days and for PR and CR, it was declined to 0.357 and 68.71% in critical HSZ., Conclusion: The negative effect of THI was observed on EBV of fertility traits under the non-HSZ and critical HSZ Thus, the influence of THI should be adjusted before estimating the breeding values for fertility traits in Murrah buffaloes.

Lactoferrin (Lf) gene promoter was screened for the presence of single nucleotide polymphism in indigenous and crossbred cattle from North India and to evaluate its association with Mastitis. Study revealed the presence of genetic variation in regulatory region of bovine Lactoferrin gene using PCR-RFLP technique. Three genotypes namely GG, GH and HH were identified. A single nucleotide change, from guanine to adenine at 25th position was found to be significantly associated (p<0.05) with clinical mastitis in indigenous Sahiwal and crossbred Karan Fries cattle maintained at organised herd of National Dairy Research Institute, Karnal. A non-significant association was observed between subclinical mastitis, somatic cell score (SCS), and GG genotype in Karan Fries cattle, however, a lower SCS was observed in animals having GG genotype. Overall a lower incidence of clinical mastitis was recorded in those animals having GG genotype of Lf in Sahiwal and Karan Fries (KF) cattle. The SNP identified in the promoter region may effect expression lactoferrin protein, which may lead to different levels of antibacterial and anti-inflammatory activity of Lf gene. Results from this study indicated the probable role played by Lactoferrin promoter to serve as candidate gene for mastitis susceptibility among indigenous and crossbred milch cattle.

Sub-fertility is a major problem in crossbred bulls leading to disintegration of breeding systems and huge economic loss. Identification of some potential biomarkers to determine the latent fertility of bulls accurately has long been the interest of researchers. In this study, we analyzed the proteome of seminal plasma (SP) from bulls with varying fertility to identify the fertility-associated proteins. The proteomic profile of high- and low-fertile bulls was compared by two-dimensional difference gel electrophoresis and differentially expressed proteins were identified through matrix-assisted laser desorption/ionization-time of flight/mass spectrometry. Out of the 18 differentially expressed proteins (P < 0.05), 9 were overexpressed in SP of high-fertile bulls and 9 were overexpressed in SP of low-fertile bulls. The differential expressions ranged from 1.5- to 5.5-fold between the two groups, where protection of telomeres-1 protein (POT1) was highly overexpressed (2.9-fold) in high-fertile group and prostaglandin E2 receptor EP3 (PTGER3) was highly abundant (5.5-fold) in low-fertile group. The protein interaction network was elucidated using STRING software tool, and the functional bioinformatics analysis was done using Blast2Go software. Most of the differentially expressed proteins were found to be involved in cellular processes and biological regulation with binding and catalytic function. It is inferred that the expression of certain proteins in the SP varied with bull fertility, and concurrent appraisal of their expression along with other fertility assays may help in determining bull fertility., (Copyright © 2014 Elsevier Inc. All rights reserved.)

The DNA methylation pattern in spermatozoa of buffalo bulls of different fertility status was investigated. Spermatozoa isolated DNA from two groups of buffalo bulls (n = 5), selected based on their artificial insemination-generated conception rate data followed by IVF efficiency, were studied for global methylation changes using a custom-designed 180 K buffalo (Bubalus bubalis) CpG island/promoter microarray. A total of 96 individual genes with another 55 genes covered under CpG islands were found differentially methylated in sperm of high-fertile and subfertile buffalo bulls. Important genes associated with biological processes, cellular components, and functions were identified to be differentially methylated in buffalo bulls with differential fertility status. The identified differentially methylated genes were found to be involved in germ cell development, spermatogenesis, capacitation, and embryonic development. The observations hint that methylation defects of sperm DNA may play a crucial role in determining the fertility of breeding bulls. This growing field of sperm epigenetics will be of great benefit in understanding the graded fertility conditions of breeding bulls in commercial livestock production system., (Copyright © 2014 Elsevier Inc. All rights reserved.)

Selection for genetically superior Murrah buffaloes under Network Project on Buffalo Improvement, India, is presently based on milk yield, and it was observed that even in the absence of any direct selection pressure applied on fertility, there has been a downward trend in fertility associated with the selection for milk yield. The aim of the study was to develop selection indices which include fertility besides milk yield in Murrah buffaloes. Data pertaining to 1,224 lactation records spread over a period of 19 years were recorded and analyzed in the study. The negative association of pregnancy rate (fertility) with a 305-day milk yield (-0.08 ± 0.04) and wet average (-0.12 ± 0.02) indicated the importance of incorporating fertility trait in the construction of selection index. Four types of selection indices were constructed and evaluated. Two indices were developed using expected producing ability 305-days milk yield (EPAMY) with 6.5 and 4% fat in milk along with expected fertilizing ability (EFA). The other two indices developed consist of expected producing ability wet average (EPA WA) and EFA. The index involving (EPA WA and EFA) was found to be a more effective selection criterion in our herd, as the accuracy of index was more (0.61), in comparison to the index involving EPA MY and EFA. The robustness of selection index was assessed by increasing the relative economic values of included traits by 25 and 50%, and accuracy of the index remains almost stable without much change. The developed selection strategy involving EPA WA and EFA should be considered for the genetic evaluation of Murrah buffaloes, as it has a potential for maximizing the lifetime reproduction and production performances of the breed.

tRNA restriction by anticodon nucleases underlies cellular stress responses and self-nonself discrimination in a wide range of taxa. Anticodon breakage inhibits protein synthesis, which, in turn, results in growth arrest or cell death. The eukaryal ribotoxin PaT secreted by Pichia acaciae inhibits growth of Saccharomyces cerevisiae via cleavage of tRNA(Gln(UUG)). We find that recombinant PaT incises a synthetic tRNA(Gln(UUG)) stem-loop RNA by transesterification at a single site 3' of the wobble uridine, yielding 2',3'-cyclic phosphate and 5'-OH ends. Incision is suppressed by replacement of the wobble nucleobase with adenine or guanine. The crystal structure of PaT reveals a distinctive fold and active site, essential components of which are demonstrated by mutagenesis. Pichia acaciae evades self-toxicity via a distinctive intracellular immunity protein, ImmPaT, which binds PaT and blocks nuclease activity. Our results highlight the evolutionary diversity of tRNA restriction and immunity systems., (Copyright © 2014 The Authors. Published by Elsevier Inc. All rights reserved.)

The decline in the male reproductive ability in terms of sexual behavior and seminal traits might lead to nonavailability of required number of bulls in a progeny testing program. The present study was conducted in 493 crossbred cattle (Bos taurus × Bos indicus) bulls to study polymorphisms of growth hormone (GH) gene and its association with seminal and sexual behavioral characteristics. A 428-base pair fragment of GH gene spanning over the fourth exon, fourth intron, and fifth exon was amplified and digested with AluI restriction enzyme. Bulls were found to be polymorphic, with two variants, LL and LV, and higher genotypic frequency for LL being 0.88. Twelve nucleotide changes and a single nonsynonymous substitution of Leucine by Valine were observed from GH1 (L) to GH2 (V). Statistical analysis revealed that the genotype of the GH gene had a significant effect on libido score, reaction time, Flehmen response, requirement of mounting stimulus, sperm mass activity, number of semen doses per collection, individual fresh sperm motility, postthaw sperm motility, acrosome integrity, hypo-osmotic swelling test, live and dead count, total morphological abnormality, and head abnormality of sperm in crossbred bulls. Growth hormone gene might be considered a candidate gene for seminal and sexual behavioral traits in crossbred cattle., (Copyright © 2014 Elsevier Inc. All rights reserved.)

A predictive model for daughter pregnancy rate (DPR) in Indian Murrah buffaloes has been developed. The data pertaining to first lactation reproduction traits, viz., age at first calving (AFC), first service period (FSP), and number of services per first conception (NS/FCON) of 719 Murrah buffaloes calved from 1985 to 2010 at the National Dairy Research Institute (NDRI), Karnal (Haryana), India were adjusted against significant environmental influence. First lactation reproduction records of 474 Murrah buffaloes were used for determining voluntary waiting period (VWP) and estimating DPR. Seven simple and multiple regression models in each case were developed where the buffaloes had their first insemination after 63 days (DPR 63), 84 days (DPR 84), and 105 days (DPR 105) of first calving. Among the seven models (I to VII) for DPR 63, DPR 84, and DPR 105, model II, having only FSP as an independent reproduction variable, was found to be the best prediction model, based on the four statistical criterions (high coefficient of determination (R 2), low mean sum of squares due to error (MSSe), conceptual predictive (CP) value, and Bayesian information criterion (BIC)). Three linear equations were developed using model II, viz., DPR 63 = 0.0033 (274 − FSP), DPR 84 = 0.0027 (332 − FSP), and DPR 105 = 0.0027 (310 − FSP). The average errors for the prediction of DPR 63, DPR 84, and DPR 105 were 1, 13, and 8 %, respectively. Therefore, 63 days of VWP is the optimum period for getting the best DPR in Indian Murrah buffaloes.

The study was conducted to assess early-expressed reproductive traits of Boran cattle and their crosses with Jersey and Holstein Friesian (HF). The traits studied were age at first services (AFS), number of services for first conception, age at first calving (AFC), first dry period (FDP), first calving interval (FCI), and first service period (FSP). Genetic group and period of birth/calving had a significant (p < 0.05) effect on reproductive traits. The Boran cattle were inferior to HF or Jersey crosses. First crosses (F 1) for Jersey and Boran (50 % Jersey: 50 % Boran) showed a significantly (p < 0.05) younger AFS (by 7.25 months) and AFC (by 10.75 months), had shorter FCI (by 63.27 days), FDP (by 61.13 days), and FSP (by 60.3 days), and needed less (by 0.35) numbers of services per first conception as compared to the Boran cattle. The F 1 for Jersey and Boran (50 % Jersey: 50 % Boran) crosses showed better performance than the F 1 for HF and Boran (50 % HF: 50 % Boran). Heritability values for AFS and AFC were the highest and were estimated at 0.51 ± 0.10 and 0.49 ± 0.13, respectively, and lowest heritability was recorded for FDP (0.02 ± 020) and FSP (0.10 ± 0.29). The genetic correlation was highest (0.10 ± 0.20) between AFS and AFC and was lowest (-0.01 ± 0.66) between FCI and FSP. The breed additive for Jersey was only significant (p < 0.01) for AFS and AFC. The crossing of HF with Boran cattle has desirably reduced 9.16 ± 2.88 months in AFS; the corresponding reduction in AFS was 3.49 ± 3.59 months by crossing with Jersey. The performance comparisons and genetic and crossbreeding parameters indicated that crossbreeding of Boran with HF or Jersey can improve reproductive performance.

There are many biological contexts in which DNA damage generates "dirty" breaks with 3'-PO4 (or cyclic-PO4) and 5'-OH ends that cannot be sealed by DNA ligases. Here we show that the Escherichia coli RNA ligase RtcB can splice these dirty DNA ends via a unique chemical mechanism. RtcB transfers GMP from a covalent RtcB-GMP intermediate to a DNA 3'-PO4 to form a "capped" 3' end structure, DNA3'pp5'G. When a suitable DNA 5'-OH end is available, RtcB catalyzes attack of the 5'-OH on DNA3'pp5'G to form a 3'-5' phosphodiester splice junction. Our findings unveil an enzymatic capacity for DNA 3' capping and the sealing of DNA breaks with 3'-PO4 and 5'-OH termini, with implications for DNA repair and DNA rearrangements.

The RNA ligase RtcB splices broken RNAs with 5'-OH and either 2',3'-cyclic phosphate or 3'-phosphate ends. The 3'-phosphate ligase activity requires GTP and entails the formation of covalent RtcB-(histidinyl)-GMP and polynucleotide-(3')pp(5')G intermediates. There are currently two models for how RtcB executes the strand sealing step. Scheme 1 holds that the RNA 5'-OH end attacks the 3'-phosphorus of the N(3')pp(5')G end to form a 3',5'-phosphodiester and release GMP. Scheme 2 posits that the N(3')pp(5')G end is converted to a 2',3'-cyclic phosphodiester, which is then attacked directly by the 5'-OH RNA end to form a 3',5'-phosphodiester. Here we show that the sealing of a 2',3'-cyclic phosphate end by RtcB requires GTP, is contingent on formation of the RtcB-GMP adduct, and involves a kinetically valid RNA(3')pp(5')G intermediate. Moreover, we find that RtcB catalyzes the hydrolysis of a 2',3'-cyclic phosphate to a 3'-phosphate at a rate that is at least as fast as the rate of ligation. These results weigh in favor of scheme 1. The cyclic phosphodiesterase activity of RtcB depends on GTP and the formation of the RtcB-GMP adduct, signifying that RtcB guanylylation precedes the cyclic phosphodiesterase and 3'-phosphate ligase steps of the RNA splicing pathway.

A cherished tenet of nucleic acid enzymology holds that synthesis of polynucleotide 3'-5' phosphodiesters proceeds via the attack of a 3'-OH on a high-energy 5' phosphoanhydride: either a nucleoside 5'-triphosphate in the case of RNA/DNA polymerases or an adenylylated intermediate A(5')pp(5')N--in the case of polynucleotide ligases. RtcB exemplifies a family of RNA ligases implicated in tRNA splicing and repair. Unlike classic ligases, RtcB seals broken RNAs with 3'-phosphate and 5'-OH ends. Here we show that RtcB executes a three-step ligation pathway entailing (i) reaction of His337 of the enzyme with GTP to form a covalent RtcB-(histidinyl-N)-GMP intermediate; (ii) transfer of guanylate to a polynucleotide 3'-phosphate to form a polynucleotide-(3')pp(5')G intermediate; and (iii) attack of a 5'-OH on the -N(3')pp(5')G end to form the splice junction. RtcB is structurally sui generis, and its chemical mechanism is unique. The wide distribution of RtcB proteins in bacteria, archaea, and metazoa raises the prospect of an alternative enzymology based on covalently activated 3' ends.

RNA 3'-phosphate cyclase (RtcA) synthesizes RNA 2',3' cyclic phosphate ends via three steps: reaction with ATP to form a covalent RtcA-(histidinyl-Nε)-AMP intermediate; transfer of adenylate to an RNA 3'-phosphate to form RNA(3')pp(5')A; and attack of the vicinal O2' on the 3'-phosphorus to form a 2',3' cyclic phosphate and release AMP. Here we report the crystal structures of RtcA•ATP, RtcA•ATP•Mn(2+), and RtcA•ATP•Co(2+) substrate complexes and an RtcA•AMP product complex. Together with the structures of RtcA apoenzyme and the covalent RtcA-AMP intermediate, they illuminate the mechanism of nucleotidyl transfer, especially the stereochemical transitions at the AMP phosphate, the critical role of the metal in orienting the PP(i) leaving group of ATP during step 1, and the protein conformational switches that accompany substrate binding and product release. The octahedral metal complex of RtcA•ATP•Mn(2+) includes nonbridging oxygens from each of the ATP phosphates, two waters, and Glu14 as the sole RtcA component. Whereas the RtcA adenylylation step is metal-catalyzed, the subsequent steps in the cyclization pathway are metal-independent.

RtcB enzymes are a newly discovered family of RNA ligases, implicated in tRNA splicing and other RNA repair reactions, that seal broken RNAs with 2',3'-cyclic phosphate and 5'-OH ends. Parsimony and energetics would suggest a one-step mechanism for RtcB sealing via attack by the O5' nucleophile on the cyclic phosphate, with expulsion of the ribose O2' and generation of a 3',5'-phosphodiester at the splice junction. Yet we find that RtcB violates Occam's razor, insofar as (i) it is adept at ligating 3'-monophosphate and 5'-OH ends; (ii) it has an intrinsic 2',3'-cyclic phosphodiesterase activity. The 2',3'-cyclic phosphodiesterase and ligase reactions both require manganese and are abolished by mutation of the RtcB active site. Thus, RtcB executes a unique two-step pathway of strand joining whereby the 2',3'-cyclic phosphodiester end is hydrolyzed to a 3'-monophosphate, which is then linked to the 5'-OH end to form the splice junction. The energy for the 3'-phosphate ligase activity is provided by GTP, which reacts with RtcB in the presence of manganese to form a covalent RtcB-guanylate adduct. This adduct is sensitive to acid and hydroxylamine but resistant to alkali, consistent with a phosphoramidate bond.

tRNA anticodon damage inflicted by the Kluyveromyces lactis γ-toxin underlies an RNA-based innate immune system that distinguishes self from nonself species. γ-toxin arrests the growth of Saccharomyces cerevisiae by incising a single phosphodiester 3' of the wobble base of tRNA(Glu(UUC)) to generate a break with 2',3'-cyclic phosphate and 5'-OH ends. Recombinant γ-toxin cleaves oligonucleotide substrates in vitro that mimic the anticodon stem-loop of tRNA(Glu). A single 2'-deoxy sugar substitution at the wobble nucleoside abolishes anticodon nuclease activity. To gain further insights to γ-toxin's substrate specificity, we tested deoxynucleoside effects at positions other than the site of transesterification. The results attest to a stringent requirement for a ribonucleoside at the uridine 5' of the wobble base. In contrast, every other nonwobble ribonucleoside in the anticodon loop can be replaced by a deoxy without significantly affecting γ-toxin's cleavage activity. Whereas either the 5' half or the 3' half of the anticodon stem can be replaced en bloc with DNA without a major effect, simultaneously replacing both strands with DNA interfered strongly, signifying that γ-toxin requires an A-form helical conformation of the anticodon stem. We purified γ-toxin mutants identified previously as nontoxic in vivo and gauged their anticodon nuclease activities in vitro. The results highlight Glu9 and Arg151 as candidate catalytic residues, along with His209 implicated previously. By analogy to other endoribonucleases, we speculate that γ-toxin drives transesterification by general acid-base catalysis (via His209 and Glu9) and transition-state stabilization (via Arg151).

RNA 3'-phosphate cyclase (Rtc) enzymes are a widely distributed family that catalyze the synthesis of RNA 2',3'-cyclic phosphate ends via an ATP-dependent pathway comprising three nucleotidyl transfer steps: reaction of Rtc with ATP to form a covalent Rtc-(histidinyl-N)-AMP intermediate and release PP(i); transfer of AMP from Rtc to an RNA 3'-phosphate to form an RNA(3')pp(5')A intermediate; and attack by the terminal nucleoside O2' on the 3'-phosphate to form an RNA 2',3'-cyclic phosphate product and release AMP. The chemical transformations of the cyclase pathway resemble those of RNA and DNA ligases, with the key distinction being that ligases covalently adenylylate 5'-phosphate ends en route to phosphodiester synthesis. Here we show that the catalytic repertoire of RNA cyclase overlaps that of ligases. We report that Escherichia coli RtcA catalyzes adenylylation of 5'-phosphate ends of DNA or RNA strands to form AppDNA and AppRNA products. The polynucleotide 5' modification reaction requires the His(309) nucleophile, signifying that it proceeds through a covalent RtcA-AMP intermediate. We established this point directly by demonstrating transfer of [(32)P]AMP from RtcA to a pDNA strand. RtcA readily adenylylated the 5'-phosphate at a 5'-PO(4)/3'-OH nick in duplex DNA but was unable to covert the nicked DNA-adenylate to a sealed phosphodiester. Our findings raise the prospect that cyclization of RNA 3'-ends might not be the only biochemical pathway in which Rtc enzymes participate; we discuss scenarios in which the 5'-adenylyltransferase of RtcA might play a role.

In present study production performance of 96 lame cows was compared with 67 healthy cows. No significant effect of parity and year of calving on milk yield were observed but the effect of season of calving was significant (P < 0.01). Effect of lameness on milk yield at the second, third and fourth months and 305 days was highly significant (P < 0.01), and was also significant (P < 0.05) on lactation yield of the fifth and tenth months. The effect of lameness on monthly and 305-day milk yield was significant (P < 0.01) only for those cows diagnosed lame before calving and during the first month of lactation. The differences in mean monthly yield were highly significant (P < 0.01) at the second, third and fourth months; and significant (P < 0.05) in the first and fifth months. The loss in the first lactation month of cows which were diagnosed as lame in the second month, was found to be significant (P < 0.05). Thus the yield of the month previous to the diagnosis (sub-clinical stage) was also affected. A significant (P < 0.01) total loss of 498.95 kg of milk yield was observed during a period of 305 days., (© 2010 The Authors. Journal compilation © 2010 Japanese Society of Animal Science.)

Search for a novel anti-inflammatory agent from a herbal source, such as Eupatorium adenophorum Spreng., a plant from the Eastern Himalayas, is of prime interest in the present investigation. Inflammation causes tissue destruction and development of diseases such as asthma, rheumatoid arthritis, and so forth. The ethanolic leaf extract of E. adenophorum (EEA) was administered intravenously and in other cases topically at the site of delayed type hypersensitivity (DTH) reaction in mouse foot paw induced with dinitrofluorobenzene. EEA can effectively inhibit DTH reaction and bring back normalcy to the paw much earlier than the controls. Efficacy of EEA on regulatory mechanisms for inflammation has also been considered. Intravenous administration of EEA increased the number of CD4(+) T cells in spleen and tumor necrosis factor (TNF)-α in serum of DTH mice. Initially it was difficult to reconcile with the anti-inflammatory role of EEA and simultaneous induction of TNF-α, an established pro-inflammatory cytokine. EEA induces higher expression of TNF-α gene and amount of the cytokine in serum. We discussed the other role of TNF-α, its involvement in repairing tissue damage incurred in course of inflammatory reaction. EEA also induces TGF-β encoding a cytokine involved in tissue repair mechanism. EEA inhibits expression of another pro-inflammatory cytokine gene IL-1β and downregulates cycloxygenase 2 (COX2) gene responsible for metabolism of inflammatory mediators like prostaglandins. Furthermore, anti-inflammatory role of EEA is also revealed through its inhibition of hydroxyl radical generation. Notably EEA does not necessarily affect the expression of other inflammation-related genes such as IL-6, IL-10 and IKK. The present study reports and analyzes for the first time the anti-inflammatory property of the leaf extract of E. adenophorum.

CD14 is an important molecule for innate immunity that can act against a wide range of pathogens. The present paper has characterized CD14 gene of crossbred (CB) cattle (Bos indicus×Bos taurus). Cloning and sequence analysis of CD14 cDNA revealed 1119 nucleotide long open reading frame encoding 373 amino acids protein and 20 amino acids signal peptide. CB cattle CD14 gene exhibited a high percentage of nucleotide identity (59.3-98.1%) with the corresponding mammalian homologs. Cattle and buffalo appear to have diverged from a common ancestor in phylogenetic analysis. 25 SNPs with 17 amino acid changes were newly reported and the site for mutational hot-spot was detected in CB cattle CD14 gene. Non-synonymous substitutions exceeding synonymous substitutions indicate the evolution of this protein through positive selection among domestic animals. Predicted protein structures obtained from deduced amino acid sequence indicated CB cattle CD14 molecule to be a receptor with horse shoe-shaped structure. The sites for LPS binding, LPS signalling, leucine-rich repeats, putative N-linked glycosylation, O-linked glycosylation, glycosyl phosphatidyl inositol anchor, disulphide bridges, alpha helix, beta strand, leucine rich nuclear export signal, leucine zipper and domain linker were predicted. Most of leucine and cysteine residues remain conserved across the species.

Natural killer (NK) cells are spontaneously cytotoxic against tumour target cells. Their number was found to be four times more in the spleen of tumour-bearing Swiss albino mice. After activation with recombinant interleukin-2 (rIL-2), NK cells were tested and found to seek out the tumour site when injected intravenously in tumour-bearing mice. Their potential for fighting tumours in vivo was further seen following adoptive transfer of rIL-2 activated NK (A-NK) cells in tumour-bearing mice. After surgical removal of tumour load, adoptive transfer of A-NK cells inhibited tumour recurrence in 92.3%cases, thereby suggesting the use of this protocol for therapeutic purposes to obtain a better outcome.

Three steroidal saponins, racemosides A (1), B (2) and C (3), were isolated from the methanolic extract of the fruits of Asparagus racemosus, and characterized as (25S)-5beta-spirostan-3beta-ol-3-O-{beta-D- glucopyranosyl (1-->6)-[alpha-L-rhamnopyranosyl (1-->6)-beta-D-glucopyranosyl (1-->4)]-beta-D-glucopyranoside}, (25S)-5beta-spirostan-3beta-ol-3-O-alpha-L-rhamnopyranosyl (1-->6)-beta-D-glucopyranosyl (1-->6)-beta-D-glucopyranoside and (25S)-5beta-spirostan-3beta-ol-3-O-{alpha-L-rhamnopyranosyl-(1-->6)-[alpha-L-rhamnopyranosyl (1-->4)]-beta-D-glucopyranoside}, respectively, by spectrometric analysis and some chemical strategies.

The present study reports the isolation and purification of lupeol acetate from the methanolic root extract of Indian medicinal plant Hemidesmus indicus (L.) R.Br. (family: Asclepiadaceae) which could neutralize venom induced action of Daboia russellii and Naja kaouthia on experimental animals. Lupeol acetate could significantly neutralize lethality, haemorrhage, defibrinogenation, edema, PLA(2) activity induced by Daboia russellii venom. It also neutralized Naja kaouthia venom induced lethality, cardiotoxicity, neurotoxicity and respiratory changes in experimental animals. Lupeol acetate potentiated the protection by snake venom antiserum action against Daboia russellii venom induced lethality in male albino mice. Venom induced changes in lipid peroxidation and super oxide dismutase activity was antagonized by lupeol acetate. Snake venom neutralization by lupeol acetate and its possible mechanism of action has been discussed.

Pathophysiology due to snakebite is a combined effect of various actions of the complex venom constituents. Importance of protein toxins in snake envenomation is well known. The present investigation reports the existence of nonprotein/nonpetide low molecular weight toxin in Indian King Cobra venom, which plays an important role in envenomation consequences in experimental animal models. A group of non-peptidic toxins (OH-NPT1) was isolated from Indian King Cobra Ophiophagus hannah by thin layer chromatography and silica gel column chromatography. UV, IR, NMR and (ESI) TOF-MS studies characterized the OH-NPT1 as a mixture of aliphatic acids having molecular weights 256, 326 and 340Da. The minimum lethal dose of OH-NPT1 was found to be 2.5 microg/20g (iv) and 4microg/20g (ip) in male albino mice. The cardiotoxic property of OH-NPT1 was established through studies on isolated guinea pig heart and auricle preparations, ECG studies in albino rat and estimation of LDH1/LDH and CPK-MB/CPK ratio in Swiss albino mice. Commercial antiserum failed to neutralize the lethality and cardiotoxicity of the toxin. However, calcium and magnesium effectively neutralized the lethal action.