Abstract The primary effluent of Indian dairy industries, cheese whey, and its main component lac... more Abstract The primary effluent of Indian dairy industries, cheese whey, and its main component lactose is considered an environmental pollutant due to its high chemical and biological oxygen demand. The model fermentative yeast, Saccharomyces cerevisiae, does not possess the necessary enzyme to utilize lactose in whey. In the present study, metabolic engineering of wild type S. cerevisiae strain with CRISPR Cas9 system was used to construct a cell factory that can flux higher galactose from the concentrated whey medium. The improved utilization of galactose confers fermentative metabolism with improved ethanol production rate. The strain provided a high titer of the ethanol (0.32±0.007 g/g) with improved galactose flux from concentrated whey at the expense of the biomass, glycerol, and acetic acid. Our present study provides evidence that galactose to ethanol fermentation is a feasible alternative for industrial waste stream treatment in developing countries.
The innate immune system is activated upon virus invasion of a host cell by recognizing viral com... more The innate immune system is activated upon virus invasion of a host cell by recognizing viral component, such as dsRNA through specific receptors, resulting in the production of type- I IFNs, which confer an antiviral state within the invaded as well as surrounding cells. In the present study, fibroblast, monocyte and macrophage cells derived from water Buffalo (Bubalus bubalis) were exposed to a synthetic dsRNA analogue, poly I:C to mimic viral invasion in each cell type. Recognition of poly I:C through cytosolic helicase receptors RIG-I and MDA5 molecule lead to the activation of the RLR pathway, subsequently activating the MAVS-IRF3/7 cascade and the production of antiviral effector molecule like IFNβ and ISGs. Within the different cell types, we identified variability in RLR receptor and IFNβ expression after poly I:C administration. Fibroblasts responded quickly and strongly with IFNβ production, followed by macrophages and monocytes. Despite absolute expression variability among different cell types the expression trend of RLRs pathway genes were similar. Length of poly I:C molecule also influence IFNβ expression in response of RLR pathway. Short (LMW) poly I:C induce stronger IFN-β expression in myeloid (macrophage and monocyte) cells. In contrast long (HMW) poly I:C preferably elicit higher IFNβ expression in non-myeloid (fibroblast) cell. Therefore, MDA5 and RIG-1 plays an indispensable role in eliciting antiviral response in non- immune (fibroblast) host cell. Thus, stimulation of RLR pathway with suitable and potentially cell-type specific agonist molecules successfully elicit antiviral state in the host animal, with fibroblasts conferring a stronger antiviral state compared with the monocytes and macrophages.
Anti- Mullerian hormone (AMH) appears to be the best biochemical-endocrine marker in assessing th... more Anti- Mullerian hormone (AMH) appears to be the best biochemical-endocrine marker in assessing the age-related decline of the ovarian follicular reserve. There is inadequate information on age-related changes in peripheral AMH concentrations in goats (Capra hircus). The present study thus aimed (i) to investigate the responsiveness of the ovary to gonadotrophin releasing hormone (GnRH) challenge to release plasma AMH into circulation in view of addressing the question whether ovarian secretion of AMH would be affected by gonadotrophic status in goats and (ii) to determine plasma AMH concentrations at different ages in goats (n= 82) using enzyme immune-assay technique. Data on hormonal concentrations in GnRH-treated animals as well as in different ages of animals were analyzed by nonparametric one-way repeated measure ANOVA and one-way ANOVA, respectively. There was no change (P greater than 0.05) of plasma AMH concentrations after GnRH challenge in experimental goats. Plasma AMH concentrations changed significantly (P less than 0.05) between the mean rank of the related groups over the time (age) in goats. Plasma AMH concentration at 3 months of age was 0.36 ± 0.04 ng/ ml and thereafter it started increasing and reached³ 2 ng/ ml between 3 and 4 years of age and then showed a progressive decline from 5 years of age and reached at 0.30 ± 0.07 ng/ ml at 10 years of age. A cubic model was found to be the best fitted model that delineated the change of plasma AMH level with age. Hence, the present study indicated that peripheral AMH concentration could be a useful management tool for the selection of goats in breeding programmes.
Background: ALRs (AIM2-like Receptors) are germline encoded PRRs that belong to PYHIN gene family... more Background: ALRs (AIM2-like Receptors) are germline encoded PRRs that belong to PYHIN gene family of cytokines, which are having signature N-terminal PYD (Pyrin, PAAD or DAPIN) domain and C-terminal HIN-200 (hematopoietic, interferon-inducible nuclear protein with 200 amino acid repeat) domain joined by a linker region. The positively charged HIN-200 domain senses and binds with negatively charged phosphate groups of single-stranded DNA (ssDNA) and double-stranded DNA (dsDNA) purely through electrostatic attractions. On the other hand, PYD domain interacts homotypically with a PYD domain of other mediators to pass the signals to effector molecules downwards the pathways for inflammatory responses. There is remarkable inter-specific diversity in the numbers of functional PYHIN genes e.g. one in cow, five in human, thirteen in mice etc., while there is a unique loss of PYHIN genes in the bat genomes which was revealed by Ahn et al. (2016) by studying genomes of ten different bat species belonging to suborders yinpterochiroptera and yangochiroptera. The conflicts between host and pathogen interfaces are compared with "Red queen's arms race" which is also described as binding seeking dynamics and binding avoidance dynamics. As a result of this never-ending rivalry, eukaryotes developed PRRs as antiviral mechanism while viruses developed counter mechanisms to evade host immune defense. The PYHIN receptors are directly engaged with pathogenic molecules, so these should have evolved under the influence of selection pressures. In the current study, we investigated the nature of selection pressure on different domain types of IFI16-like (IFI16-L) PYHIN genes in ruminants. Results: Three transcript variants of the IFI16-like gene were found in PBMCs of ruminant animals-water buffalo, zebu cattle, goat, and sheep. The IFI16-like gene has one N-terminal PYD domain and one C-terminal HIN-200 domain, separated by an inter-domain linker region. HIN domain and inter-domain region are positively selected while the PYD domain is under the influence of purifying selection. Conclusion: Herein, we conclude that the nature of selection pressure varies on different parts (PYD domain, HIN domain, and inter-domain linker region) of IFI16-like PYHIN genes in the ruminants. This data can be useful to predict the molecular determinants of pathogen interactions.
SummaryMaturing oocytes have diverse developmental potential and good quality oocytes exhibit a b... more SummaryMaturing oocytes have diverse developmental potential and good quality oocytes exhibit a better ability to attain physiological milestones in a time-dependent manner. This situation necessitates the confirmation of oocyte developmental status more precisely under an in vitro embryo production (IVEP) regime. The aim of this study was to explain timely events in germinal vesicle breakdown (GVBD), an important milestone of oocyte nuclear maturation, to delineate the developmental capacity of Bubalus bubalis oocytes. In addition, the expression profile of genes responsible for GVBD was assessed in order to understand the molecular context responsible for GVBD. The chronology of GVBD events at different time intervals during in vitro maturation (IVM) suggests that the rate at which oocytes undergo GVBD was strikingly different in the brilliant cresyl blue (BCB)+ and BCB- groups. The expression of AKT and CDC25B genes for BCB+ oocytes was maximum at 8 h of IVM, and CCNB (cyclin B) ...
The present study was conceived with the aim of modulating the cumulus expansion characteristics ... more The present study was conceived with the aim of modulating the cumulus expansion characteristics of poor quality (BCB-) buffalo oocyte complexes (COCs) in order to improve their fertilization outcomes. BCB- COCs were subjected to in vitro maturation (IVM) in presence of Interleukin - 1 beta (IL-1β) along with BCB- (control) and good quality (BCB +) COCs. Results were assessed morphologically, by scanning electron microscopy (SEM) and by expression analysis of cumulus expansion related genes. Also, numbers of zona pellucida bound spermatozoa were counted and development rates of oocytes were monitored under different groups. Expression of versican isoforms and ADAMTS-1 was observed to be significantly different between cumulus cells of BCB+ and BCB- COCs. Upon IL-1β supplementation, ADAMTS-1 expression increased in BCB- COCs along with corresponding cumulus expansion rates. SEM analysis also revealed improved cumulus expansion in IL-1β supplemented BCB- COCs. HAS2 and TNFAIP-6 were s...
Germinal vesicle breakdown (GVBD) is the first milestone that an oocyte needs to achieve towards ... more Germinal vesicle breakdown (GVBD) is the first milestone that an oocyte needs to achieve towards completing the maturation and gaining potential to fertilise. Significantly lower in vitro embryo production rate in buffaloes can be attributed to heterogeneity of GVBD occurrence among oocytes obtained from abattoir derived ovaries. Evidence from our earlier work had suggested that different qualities of buffalo oocytes differ significantly in their timing of GVBD. Besides, these oocytes also differ in terms of volume of Akt phosphorylation, which initiates the process of GVBD. With objective of synchronizing the oocytes for GVBD, immature buffalo oocytes were subjected to a two-step culture protocol, initially in the presence of GVBD inhibitors and subsequently, in vitro maturation (IVM) with added SC79 (activates Akt). Expression of developmentally important genes was assessed along with embryo development rate and blastocyst health to interpret the consequences. Oocytes subjected to...
Cover: The cover image, by Tirtha K. Datta et al., is based on the Article Differential Expressio... more Cover: The cover image, by Tirtha K. Datta et al., is based on the Article Differential Expression of Newly Identified Long Intergenic Non-Coding RNAs in Buffalo Oocytes Indicating Their Possible Role in Maturation and Embryonic Development, DOI: 10.1002/jcp.25828.
Journal of molecular recognition : JMR, Sep 3, 2017
Inflammasomes are multiprotein caspase-activating complexes that enhance the maturation and relea... more Inflammasomes are multiprotein caspase-activating complexes that enhance the maturation and release of proinflammatory cytokines (IL-1β and IL-18) in response to the invading pathogen and/or host-derived cellular stress. These are assembled by the sensory proteins (viz NLRC4, NLRP1, NLRP3, and AIM-2), adaptor protein (ASC), and effector molecule procaspase-1. In NLRP3-mediated inflammasome activation, ASC acts as a mediator between NLRP3 and procaspase-1 for the transmission of signals. A series of homotypic protein-protein interactions (NLRP3(PYD) :ASC(PYD) and ASC(CARD) :CASP1(CARD) ) propagates the downstream signaling for the production of proinflammatory cytokines. Pyrin-only protein 1 (POP1) is known to act as the regulator of inflammasome. It modulates the ASC-mediated inflammasome assembly by interacting with pyrin domain (PYD) of ASC. However, despite similar electrostatic surface potential, the interaction of POP1 with NLRP3(PYD) is obscured till date. Herein, to explore t...
Hybrid sterility or reproductive isolation in mammals has been attributed to allelic incompatibil... more Hybrid sterility or reproductive isolation in mammals has been attributed to allelic incompatibilities in a DNA-binding protein PRDM9. Not only is PRDM9 exceptional in being the only known 'speciation gene' in vertebrates, but it is also considered to be the fastest evolving gene in the genome. The terminal zinc finger (ZF) domain of PRDM9 specifies genome-wide meiotic recombination hotspot locations in mammals. Intriguingly, PRDM9 ZF domain is highly variable between as well as within species, possibly activating different recombination hotspots. The present study characterized the full-length coding sequence of PRDM9 in cattle and buffalo and explored the diversity of the ZF array in 514 samples from different bovids (cattle, yak, mithun, and buffalo). Substantial numerical and sequence variability were observed in the ZFs, with the number of repeats ranging from 6 to 9 in different bovines. Sequence analysis revealed the presence of 37 different ZFs in cattle, 3 in mithun, 4 in yak, and 13 in buffaloes producing 41 unique PRDM9 alleles in these species. The posterior mean of dN/dS or omega values calculated using Codeml tool of PAMLX identified sites -5, -1, +2, +3, +4, +5, and +6 in the ZF domain to be evolving positively in the studied species. Concerted evolution which typifies the evolution of this gene was consistently evident in all bovines. Our results demonstrate the extraordinary diversity of PRDM9 ZF array across bovines, reinforcing similar observations in other metazoans. The high variability is suggestive of unique repertoire of meiotic recombination hotspots in each species.
Somatic Cell Count (SCC) is an indicator to detect mastitis in individual cows. The SCC was measu... more Somatic Cell Count (SCC) is an indicator to detect mastitis in individual cows. The SCC was measured from individual milk samples of 275 animals comprising of 164 HF crossbreds, 51 Deoni and 60 Ongole breeds of cattle. The mean milk yield of Deoni, Ongole and HF crossbred cows was 824.44 ± 149.99, 479.73 ± 141.14 and 5572.15 ± 115.28 kg, respectively. The mean SCC in Deoni, Ongole and HF crossbred cows were 1.95 ± 0.24, 1.57 ± 0.22 and 4.14 ± 0.17 lakhs cells per ml of milk, respectively. Crossbred cows showed significantly (Plactation were significant (P on stage of lactation on SCC. The changes in SCC during different seasons were significant (pre as in Ongole the SCC was high at lean season. SCC was increased with increasing number of lactation in Deoni cows. The highest SCC value (18.56 ± 1.11 x105 cells/ml) was observed at 4th parity in HF mastitis animals.
Neutrophils being the first line of cellular defense show significant difference in their express... more Neutrophils being the first line of cellular defense show significant difference in their expression in pregnant (P) and non-pregnant (NP) cows during the peri-implantation period. To study these changes, blood samples were collected from cows coming in heat and brought for artificial insemination (AI) on day 0 (day of AI), 4, 8, 12, 14, 16, 18, 21, 24, 30 and 40day post-AI. Pregnancy was confirmed by observing non-return of heat, progesterone assay and ultrasonography. Cows were then categorized into pregnant (n=10) and non-pregnant (n=10) groups. Blood neutrophils were isolated and plasma samples collected from both P and NP cows during this period. RNA from neutrophils was isolated and studied for gene expression of adhesion molecules (CD62L, CD11b), chemokine (IL-8), and interferon stimulated genes (ISG15, OAS1, MX1, MX2, IFI16 and IFI44) using qRT-PCR. Adhesion molecules along with IL-8 showed a higher expression in NP cows. Expression of IFI16 was up-regulated as early as day ...
Riboflavin has an important role in various cellular metabolic activities through its participati... more Riboflavin has an important role in various cellular metabolic activities through its participation in oxidation-reduction reactions. In this study, as many as 60 lactobacilli were screened for the presence or absence of riboflavin biosynthesis genes and riboflavin production. Of these, only 14 strains were able to grow in a commercial riboflavin-free medium. We observed that the presence of riboflavin biosynthesis genes is strain-specific across different species of lactobacilli. The microbiological assay was found to be appreciably reproducible, sensitive, rapid, and inexpensive and, hence, can be employed for screening the riboflavin-producing strains. The study thus represents a convenient and efficient method for selection of novel riboflavin producers. These riboflavin(+) strains thus identified and characterized could be explored as potent candidates for the development of a wide range of dairy- and cereal-based foods for the delivery of in situ riboflavin to consumers.
Consumers are increasingly becoming aware of their health and nutritional requirements, and in th... more Consumers are increasingly becoming aware of their health and nutritional requirements, and in this context, vitamins produced in situ by microbes may suit their needs and expectations. B groups vitamins are essential components of cellular metabolism and among them riboflavin is one of the vital vitamins required by bacteria, plants, animals and humans. Here, we focus on the importance of microbial production of riboflavin over chemical synthesis. In addition, genetic abilities for riboflavin biosynthesis by lactic acid bacteria are discussed. Genetically modified strains by employing genetic engineering and chemical analogues have been developed to enhance riboflavin production. The present review attempts to collect the currently available information on riboflavin production by microbes in general, while placing greater emphasis on food grade lactic acid bacteria and human gut commensals. For designing riboflavin-enriched functional foods, proper selection and exploitation of riboflavin-producing lactic acid bacteria is essential. Moreover, eliminating the in situ vitamin fortification step will decrease the cost of food production.
Cathelicidins are an ancient class of antimicrobial peptides (AMPs) with broad spectrum bacterici... more Cathelicidins are an ancient class of antimicrobial peptides (AMPs) with broad spectrum bactericidal activities. In this study, we investigated the diversity and biological activity of cathelicidins of buffalo, a species known for its disease resistance. A series of new homologs of cathelicidin4 (CATHL4), which were structurally diverse in their antimicrobial domain, was identified in buffalo. AMPs of newly identified buffalo CATHL4s (buCATHL4s) displayed potent antimicrobial activity against selected Gram positive (G+) and Gram negative (G-) bacteria. These peptides were prompt to disrupt the membrane integrity of bacteria and induced specific changes such as blebing, budding, and pore like structure formation on bacterial membrane. The peptides assumed different secondary structure conformations in aqueous and membrane-mimicking environments. Simulation studies suggested that the amphipathic design of buCATHL4 was crucial for water permeation following membrane disruption. A great...
Buffalo (Bubalus bubalis) is an important animal species for small and marginal farmers involved ... more Buffalo (Bubalus bubalis) is an important animal species for small and marginal farmers involved in farming. Buffalo farming is also important for the growing economy of India and many other countries worldwide. Buffaloes are used either as draught animals or as a source of meat, horns, skin, milk and milk products. The genomic information available in public domain needs to be organized in a systematic manner for knowledge discovery. The aim of this paper is to develop a database on buffalo genome resources orthologous to Cattle (Bos taurus). Initially, the information on functional elements of Bos taurus has been retrieved from NCBI / UCSC and then mapped on to Bubalus bubalis genome information available at NCBI. The mapped data has been parsed and a database has been populated. Also, a web-based information system has been developed using LAMP technology. For better visualization, a genome browser has been developed based on light weight genome viewer tool for displaying the map...
Abstract The primary effluent of Indian dairy industries, cheese whey, and its main component lac... more Abstract The primary effluent of Indian dairy industries, cheese whey, and its main component lactose is considered an environmental pollutant due to its high chemical and biological oxygen demand. The model fermentative yeast, Saccharomyces cerevisiae, does not possess the necessary enzyme to utilize lactose in whey. In the present study, metabolic engineering of wild type S. cerevisiae strain with CRISPR Cas9 system was used to construct a cell factory that can flux higher galactose from the concentrated whey medium. The improved utilization of galactose confers fermentative metabolism with improved ethanol production rate. The strain provided a high titer of the ethanol (0.32±0.007 g/g) with improved galactose flux from concentrated whey at the expense of the biomass, glycerol, and acetic acid. Our present study provides evidence that galactose to ethanol fermentation is a feasible alternative for industrial waste stream treatment in developing countries.
The innate immune system is activated upon virus invasion of a host cell by recognizing viral com... more The innate immune system is activated upon virus invasion of a host cell by recognizing viral component, such as dsRNA through specific receptors, resulting in the production of type- I IFNs, which confer an antiviral state within the invaded as well as surrounding cells. In the present study, fibroblast, monocyte and macrophage cells derived from water Buffalo (Bubalus bubalis) were exposed to a synthetic dsRNA analogue, poly I:C to mimic viral invasion in each cell type. Recognition of poly I:C through cytosolic helicase receptors RIG-I and MDA5 molecule lead to the activation of the RLR pathway, subsequently activating the MAVS-IRF3/7 cascade and the production of antiviral effector molecule like IFNβ and ISGs. Within the different cell types, we identified variability in RLR receptor and IFNβ expression after poly I:C administration. Fibroblasts responded quickly and strongly with IFNβ production, followed by macrophages and monocytes. Despite absolute expression variability among different cell types the expression trend of RLRs pathway genes were similar. Length of poly I:C molecule also influence IFNβ expression in response of RLR pathway. Short (LMW) poly I:C induce stronger IFN-β expression in myeloid (macrophage and monocyte) cells. In contrast long (HMW) poly I:C preferably elicit higher IFNβ expression in non-myeloid (fibroblast) cell. Therefore, MDA5 and RIG-1 plays an indispensable role in eliciting antiviral response in non- immune (fibroblast) host cell. Thus, stimulation of RLR pathway with suitable and potentially cell-type specific agonist molecules successfully elicit antiviral state in the host animal, with fibroblasts conferring a stronger antiviral state compared with the monocytes and macrophages.
Anti- Mullerian hormone (AMH) appears to be the best biochemical-endocrine marker in assessing th... more Anti- Mullerian hormone (AMH) appears to be the best biochemical-endocrine marker in assessing the age-related decline of the ovarian follicular reserve. There is inadequate information on age-related changes in peripheral AMH concentrations in goats (Capra hircus). The present study thus aimed (i) to investigate the responsiveness of the ovary to gonadotrophin releasing hormone (GnRH) challenge to release plasma AMH into circulation in view of addressing the question whether ovarian secretion of AMH would be affected by gonadotrophic status in goats and (ii) to determine plasma AMH concentrations at different ages in goats (n= 82) using enzyme immune-assay technique. Data on hormonal concentrations in GnRH-treated animals as well as in different ages of animals were analyzed by nonparametric one-way repeated measure ANOVA and one-way ANOVA, respectively. There was no change (P greater than 0.05) of plasma AMH concentrations after GnRH challenge in experimental goats. Plasma AMH concentrations changed significantly (P less than 0.05) between the mean rank of the related groups over the time (age) in goats. Plasma AMH concentration at 3 months of age was 0.36 ± 0.04 ng/ ml and thereafter it started increasing and reached³ 2 ng/ ml between 3 and 4 years of age and then showed a progressive decline from 5 years of age and reached at 0.30 ± 0.07 ng/ ml at 10 years of age. A cubic model was found to be the best fitted model that delineated the change of plasma AMH level with age. Hence, the present study indicated that peripheral AMH concentration could be a useful management tool for the selection of goats in breeding programmes.
Background: ALRs (AIM2-like Receptors) are germline encoded PRRs that belong to PYHIN gene family... more Background: ALRs (AIM2-like Receptors) are germline encoded PRRs that belong to PYHIN gene family of cytokines, which are having signature N-terminal PYD (Pyrin, PAAD or DAPIN) domain and C-terminal HIN-200 (hematopoietic, interferon-inducible nuclear protein with 200 amino acid repeat) domain joined by a linker region. The positively charged HIN-200 domain senses and binds with negatively charged phosphate groups of single-stranded DNA (ssDNA) and double-stranded DNA (dsDNA) purely through electrostatic attractions. On the other hand, PYD domain interacts homotypically with a PYD domain of other mediators to pass the signals to effector molecules downwards the pathways for inflammatory responses. There is remarkable inter-specific diversity in the numbers of functional PYHIN genes e.g. one in cow, five in human, thirteen in mice etc., while there is a unique loss of PYHIN genes in the bat genomes which was revealed by Ahn et al. (2016) by studying genomes of ten different bat species belonging to suborders yinpterochiroptera and yangochiroptera. The conflicts between host and pathogen interfaces are compared with "Red queen's arms race" which is also described as binding seeking dynamics and binding avoidance dynamics. As a result of this never-ending rivalry, eukaryotes developed PRRs as antiviral mechanism while viruses developed counter mechanisms to evade host immune defense. The PYHIN receptors are directly engaged with pathogenic molecules, so these should have evolved under the influence of selection pressures. In the current study, we investigated the nature of selection pressure on different domain types of IFI16-like (IFI16-L) PYHIN genes in ruminants. Results: Three transcript variants of the IFI16-like gene were found in PBMCs of ruminant animals-water buffalo, zebu cattle, goat, and sheep. The IFI16-like gene has one N-terminal PYD domain and one C-terminal HIN-200 domain, separated by an inter-domain linker region. HIN domain and inter-domain region are positively selected while the PYD domain is under the influence of purifying selection. Conclusion: Herein, we conclude that the nature of selection pressure varies on different parts (PYD domain, HIN domain, and inter-domain linker region) of IFI16-like PYHIN genes in the ruminants. This data can be useful to predict the molecular determinants of pathogen interactions.
SummaryMaturing oocytes have diverse developmental potential and good quality oocytes exhibit a b... more SummaryMaturing oocytes have diverse developmental potential and good quality oocytes exhibit a better ability to attain physiological milestones in a time-dependent manner. This situation necessitates the confirmation of oocyte developmental status more precisely under an in vitro embryo production (IVEP) regime. The aim of this study was to explain timely events in germinal vesicle breakdown (GVBD), an important milestone of oocyte nuclear maturation, to delineate the developmental capacity of Bubalus bubalis oocytes. In addition, the expression profile of genes responsible for GVBD was assessed in order to understand the molecular context responsible for GVBD. The chronology of GVBD events at different time intervals during in vitro maturation (IVM) suggests that the rate at which oocytes undergo GVBD was strikingly different in the brilliant cresyl blue (BCB)+ and BCB- groups. The expression of AKT and CDC25B genes for BCB+ oocytes was maximum at 8 h of IVM, and CCNB (cyclin B) ...
The present study was conceived with the aim of modulating the cumulus expansion characteristics ... more The present study was conceived with the aim of modulating the cumulus expansion characteristics of poor quality (BCB-) buffalo oocyte complexes (COCs) in order to improve their fertilization outcomes. BCB- COCs were subjected to in vitro maturation (IVM) in presence of Interleukin - 1 beta (IL-1β) along with BCB- (control) and good quality (BCB +) COCs. Results were assessed morphologically, by scanning electron microscopy (SEM) and by expression analysis of cumulus expansion related genes. Also, numbers of zona pellucida bound spermatozoa were counted and development rates of oocytes were monitored under different groups. Expression of versican isoforms and ADAMTS-1 was observed to be significantly different between cumulus cells of BCB+ and BCB- COCs. Upon IL-1β supplementation, ADAMTS-1 expression increased in BCB- COCs along with corresponding cumulus expansion rates. SEM analysis also revealed improved cumulus expansion in IL-1β supplemented BCB- COCs. HAS2 and TNFAIP-6 were s...
Germinal vesicle breakdown (GVBD) is the first milestone that an oocyte needs to achieve towards ... more Germinal vesicle breakdown (GVBD) is the first milestone that an oocyte needs to achieve towards completing the maturation and gaining potential to fertilise. Significantly lower in vitro embryo production rate in buffaloes can be attributed to heterogeneity of GVBD occurrence among oocytes obtained from abattoir derived ovaries. Evidence from our earlier work had suggested that different qualities of buffalo oocytes differ significantly in their timing of GVBD. Besides, these oocytes also differ in terms of volume of Akt phosphorylation, which initiates the process of GVBD. With objective of synchronizing the oocytes for GVBD, immature buffalo oocytes were subjected to a two-step culture protocol, initially in the presence of GVBD inhibitors and subsequently, in vitro maturation (IVM) with added SC79 (activates Akt). Expression of developmentally important genes was assessed along with embryo development rate and blastocyst health to interpret the consequences. Oocytes subjected to...
Cover: The cover image, by Tirtha K. Datta et al., is based on the Article Differential Expressio... more Cover: The cover image, by Tirtha K. Datta et al., is based on the Article Differential Expression of Newly Identified Long Intergenic Non-Coding RNAs in Buffalo Oocytes Indicating Their Possible Role in Maturation and Embryonic Development, DOI: 10.1002/jcp.25828.
Journal of molecular recognition : JMR, Sep 3, 2017
Inflammasomes are multiprotein caspase-activating complexes that enhance the maturation and relea... more Inflammasomes are multiprotein caspase-activating complexes that enhance the maturation and release of proinflammatory cytokines (IL-1β and IL-18) in response to the invading pathogen and/or host-derived cellular stress. These are assembled by the sensory proteins (viz NLRC4, NLRP1, NLRP3, and AIM-2), adaptor protein (ASC), and effector molecule procaspase-1. In NLRP3-mediated inflammasome activation, ASC acts as a mediator between NLRP3 and procaspase-1 for the transmission of signals. A series of homotypic protein-protein interactions (NLRP3(PYD) :ASC(PYD) and ASC(CARD) :CASP1(CARD) ) propagates the downstream signaling for the production of proinflammatory cytokines. Pyrin-only protein 1 (POP1) is known to act as the regulator of inflammasome. It modulates the ASC-mediated inflammasome assembly by interacting with pyrin domain (PYD) of ASC. However, despite similar electrostatic surface potential, the interaction of POP1 with NLRP3(PYD) is obscured till date. Herein, to explore t...
Hybrid sterility or reproductive isolation in mammals has been attributed to allelic incompatibil... more Hybrid sterility or reproductive isolation in mammals has been attributed to allelic incompatibilities in a DNA-binding protein PRDM9. Not only is PRDM9 exceptional in being the only known 'speciation gene' in vertebrates, but it is also considered to be the fastest evolving gene in the genome. The terminal zinc finger (ZF) domain of PRDM9 specifies genome-wide meiotic recombination hotspot locations in mammals. Intriguingly, PRDM9 ZF domain is highly variable between as well as within species, possibly activating different recombination hotspots. The present study characterized the full-length coding sequence of PRDM9 in cattle and buffalo and explored the diversity of the ZF array in 514 samples from different bovids (cattle, yak, mithun, and buffalo). Substantial numerical and sequence variability were observed in the ZFs, with the number of repeats ranging from 6 to 9 in different bovines. Sequence analysis revealed the presence of 37 different ZFs in cattle, 3 in mithun, 4 in yak, and 13 in buffaloes producing 41 unique PRDM9 alleles in these species. The posterior mean of dN/dS or omega values calculated using Codeml tool of PAMLX identified sites -5, -1, +2, +3, +4, +5, and +6 in the ZF domain to be evolving positively in the studied species. Concerted evolution which typifies the evolution of this gene was consistently evident in all bovines. Our results demonstrate the extraordinary diversity of PRDM9 ZF array across bovines, reinforcing similar observations in other metazoans. The high variability is suggestive of unique repertoire of meiotic recombination hotspots in each species.
Somatic Cell Count (SCC) is an indicator to detect mastitis in individual cows. The SCC was measu... more Somatic Cell Count (SCC) is an indicator to detect mastitis in individual cows. The SCC was measured from individual milk samples of 275 animals comprising of 164 HF crossbreds, 51 Deoni and 60 Ongole breeds of cattle. The mean milk yield of Deoni, Ongole and HF crossbred cows was 824.44 ± 149.99, 479.73 ± 141.14 and 5572.15 ± 115.28 kg, respectively. The mean SCC in Deoni, Ongole and HF crossbred cows were 1.95 ± 0.24, 1.57 ± 0.22 and 4.14 ± 0.17 lakhs cells per ml of milk, respectively. Crossbred cows showed significantly (Plactation were significant (P on stage of lactation on SCC. The changes in SCC during different seasons were significant (pre as in Ongole the SCC was high at lean season. SCC was increased with increasing number of lactation in Deoni cows. The highest SCC value (18.56 ± 1.11 x105 cells/ml) was observed at 4th parity in HF mastitis animals.
Neutrophils being the first line of cellular defense show significant difference in their express... more Neutrophils being the first line of cellular defense show significant difference in their expression in pregnant (P) and non-pregnant (NP) cows during the peri-implantation period. To study these changes, blood samples were collected from cows coming in heat and brought for artificial insemination (AI) on day 0 (day of AI), 4, 8, 12, 14, 16, 18, 21, 24, 30 and 40day post-AI. Pregnancy was confirmed by observing non-return of heat, progesterone assay and ultrasonography. Cows were then categorized into pregnant (n=10) and non-pregnant (n=10) groups. Blood neutrophils were isolated and plasma samples collected from both P and NP cows during this period. RNA from neutrophils was isolated and studied for gene expression of adhesion molecules (CD62L, CD11b), chemokine (IL-8), and interferon stimulated genes (ISG15, OAS1, MX1, MX2, IFI16 and IFI44) using qRT-PCR. Adhesion molecules along with IL-8 showed a higher expression in NP cows. Expression of IFI16 was up-regulated as early as day ...
Riboflavin has an important role in various cellular metabolic activities through its participati... more Riboflavin has an important role in various cellular metabolic activities through its participation in oxidation-reduction reactions. In this study, as many as 60 lactobacilli were screened for the presence or absence of riboflavin biosynthesis genes and riboflavin production. Of these, only 14 strains were able to grow in a commercial riboflavin-free medium. We observed that the presence of riboflavin biosynthesis genes is strain-specific across different species of lactobacilli. The microbiological assay was found to be appreciably reproducible, sensitive, rapid, and inexpensive and, hence, can be employed for screening the riboflavin-producing strains. The study thus represents a convenient and efficient method for selection of novel riboflavin producers. These riboflavin(+) strains thus identified and characterized could be explored as potent candidates for the development of a wide range of dairy- and cereal-based foods for the delivery of in situ riboflavin to consumers.
Consumers are increasingly becoming aware of their health and nutritional requirements, and in th... more Consumers are increasingly becoming aware of their health and nutritional requirements, and in this context, vitamins produced in situ by microbes may suit their needs and expectations. B groups vitamins are essential components of cellular metabolism and among them riboflavin is one of the vital vitamins required by bacteria, plants, animals and humans. Here, we focus on the importance of microbial production of riboflavin over chemical synthesis. In addition, genetic abilities for riboflavin biosynthesis by lactic acid bacteria are discussed. Genetically modified strains by employing genetic engineering and chemical analogues have been developed to enhance riboflavin production. The present review attempts to collect the currently available information on riboflavin production by microbes in general, while placing greater emphasis on food grade lactic acid bacteria and human gut commensals. For designing riboflavin-enriched functional foods, proper selection and exploitation of riboflavin-producing lactic acid bacteria is essential. Moreover, eliminating the in situ vitamin fortification step will decrease the cost of food production.
Cathelicidins are an ancient class of antimicrobial peptides (AMPs) with broad spectrum bacterici... more Cathelicidins are an ancient class of antimicrobial peptides (AMPs) with broad spectrum bactericidal activities. In this study, we investigated the diversity and biological activity of cathelicidins of buffalo, a species known for its disease resistance. A series of new homologs of cathelicidin4 (CATHL4), which were structurally diverse in their antimicrobial domain, was identified in buffalo. AMPs of newly identified buffalo CATHL4s (buCATHL4s) displayed potent antimicrobial activity against selected Gram positive (G+) and Gram negative (G-) bacteria. These peptides were prompt to disrupt the membrane integrity of bacteria and induced specific changes such as blebing, budding, and pore like structure formation on bacterial membrane. The peptides assumed different secondary structure conformations in aqueous and membrane-mimicking environments. Simulation studies suggested that the amphipathic design of buCATHL4 was crucial for water permeation following membrane disruption. A great...
Buffalo (Bubalus bubalis) is an important animal species for small and marginal farmers involved ... more Buffalo (Bubalus bubalis) is an important animal species for small and marginal farmers involved in farming. Buffalo farming is also important for the growing economy of India and many other countries worldwide. Buffaloes are used either as draught animals or as a source of meat, horns, skin, milk and milk products. The genomic information available in public domain needs to be organized in a systematic manner for knowledge discovery. The aim of this paper is to develop a database on buffalo genome resources orthologous to Cattle (Bos taurus). Initially, the information on functional elements of Bos taurus has been retrieved from NCBI / UCSC and then mapped on to Bubalus bubalis genome information available at NCBI. The mapped data has been parsed and a database has been populated. Also, a web-based information system has been developed using LAMP technology. For better visualization, a genome browser has been developed based on light weight genome viewer tool for displaying the map...
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Papers by Sachinandan De