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Abstract: The bovine mammary gland has vital importance in the dairy sector, as it is considered a source of basic dairy product, milk. Mammary gland affections are widespread, which affect the dairy industry economically and pose a potential public health hazard. Current therapeutic options are ineffective in controlling the infection and regenerating the gland effectively. Antimicrobials commonly used against mastitis make their way into the milk . In order to find a solution to these problems, advanced therapeutic options, like the one for stem cells, are considered. Mammary gland stem cells (MaSCs) are considered to maintain tissue homeostasis. The characterization of these cells and their derived lineages (progenitor cells and mammary epithelial cells) may potentially provide the physiological status or production potential of the gland. However, current isolation methods are cumbersome and fall short to isolate a pure line of cattle MaSCs from progenitors or other differentiated epithelial cells. An alternative to the therapeutic application of MaSCs is the mesenchymal stem cell (MSC). These cells can potentially control microbial infection, show anti-inflammatory and other pro-healing effects, and furthermore enhance mammary epithelial cell secretory potential to ensure tissue regeneration. The current review focuses on MaSCs and MSCs properties in light of the bovine mammary gland regeneration.

Fetal adnexa derived Mesenchymal Stem Cells (MSC) offer all in one solution in therapeutics. The present study was designed to isolate, cultivate, characterize and to evaluate the growth kinetics of ovine fetal adnexa-derived Mesenchymal Stem Cells (MSCs). The gravid uteri of ewes were collected from a local abattoir. The four cell lines viz. Wharton’s Jelly (oWJ), Cord Blood (oCB), Amniotic Fluid (oAF), and Amniotic Sac (oAS) were isolated and expanded in vitro and characterized for surface markers (CD73, CD90 & CD105) and pluripotency markers (SOX2 & OCT4) at 3rd passage. The growth kinetics of MSCs was investigated at 3rd and 5th passages. Similarly, the colony forming unit (CFU) Assay was performed at 3rd passage. The fetal adnexa derived Ovine MSCs showed the amplification of CD73, CD90 & CD105, whereas, negative marker CD34 was not amplified. Similarly, the MSCs also qualified for the amplification of Pluripotency markers (OCT4, SOX2). The MSCs in culture showed a typical growth curve with initial lag phase, an exponential phase, a plateau phase and a decline phase. The proliferation potential of MSCs was higher at P5 than P3. The overall growth rate was significantly (p

Background: Mesenchymal Stem Cells (MSCs) are present in almost all the tissues of the body and act as the backbone of the internal tissue homeostasis. Among their various characteristic features, immuno-modulatory and/ anti-inflammatory properties play an important role in therapeutics. Objective: The current topic focuses on the characterization and immuno-modulatory and/ antiinflammatory properties of MSCs. To present and discuss the current status of MSCs immunomodulatory properties. Methods: Available literature on MSCs properties and patents have been detailed, critically interpreted, and discussed based upon available literature. The main focus has been on their characteristic immunomodulatory and anti-inflammatory properties though some of the basic characterization markers have also been detailed. The databases searched for the literature include PubMed, Med Line, PubMed Central, Science Direct and a few other scientific databases. Results: MSCs are present in a very limited concentration in the tissues, and as such their culture expansion becomes imperative. MSCs immuno-modulatory and anti-inflammatory roles are achieved through direct cell-cell contact and / by the release of certain factors. Such properties are controlled by micro-environment upon which currently very limited control can be exerted. Besides, further insights in the xeno-protein free culture media as against the fetal bovine serum is required. Conclusions: MSCs have been well-isolated, cultured and characterized from numerous tissues of the body. The majority of the studies have shown MSCs as immuno-compromised with immunomodulatory and / or anti-inflammatory properties except some of the latest studies that have failed to achieve the desired results and thus, demand further research. Further research is required in the area to translate the results into clinical application.

Mesenchymal stem cells (MSCs) due to their characteristic properties have a potential to treat osteoarthritis, one of the major growing joint problems. MSCs show differential ex vivo chondrogenic potential on the basis of source that remains to be validated under in vivo environment. This study compared chondrogenic potential of MSCs derived from two common sources, adipose tissue (AD) and bone marrow (BM) under ex vivo and in vivo environments. The randomized placebo controlled osteochondral defect (OCD) study divided n = 72 rabbits equally into Control, AD-MSCs and BM-MSCs groups. Ex vivo chondrogenic induction resulted in an increased aggrecan fold expression in BM-MSCs and AD-MSCs. The former cell type had significantly (p0.05) higher fold expression as compared to the latter. The cell treated OCDs had significantly reduced gene expression for inflammatory markers (IL-6, IL-8 and TNF-α) as compared to the control. In OCD study, radiography, MRI, gross observation, histopathology and SEM revealed that the cell treated defects were early filled by the tissue that had better surface architecture and matrices as compared to the control. BM-MSCs treated defects had better scores especially for gross and histopathology than the AD-MSCs. Gene expression for osteochondral regulation and cartilaginous matrices was higher in BM-MSCs group while only for matrices including the Col I in AD-MSCs as compared to the control. It was concluded that OCD in the cell treated groups are filled early with mostly a fibrocartilaginous to hyaline tissue. BM-MSCs may have an edge over AD-MSCs in OCD repair.

Salmonella, one of the major infectious diseases in poultry, causes considerable economic losses in terms of mortality and morbidity, especially in countries that lack effective vaccination programs. Besides being resistant to diseases, indigenous chicken breeds are also a potential source of animal protein in developing countries. For understanding the disease resistance, an indigenous chicken line Kashmir faverolla, and commercial broiler were selected. RNA-seq was performed after challenging the chicken with Salmonella Typhimurium. Comparative differential expression results showed that following infection, a total of 3153 genes and 1787 genes were differentially expressed in the liver and spleen, respectively. The genes that were differentially expressed included interleukins, cytokines, NOS2, Avβ-defensins, toll-like receptors, and other immune-related gene families. Most of the genes and signaling pathways involved in the innate and adaptive immune responses against bacterial infection were significantly enriched in the Kashmir faverolla. Pathway analysis revealed that most of the enriched pathways were MAPK signaling pathway, NOD-like receptor signaling pathway, TLR signaling pathway, PPAR signaling pathway, endocytosis, etc. Surprisingly some immune-related genes like TLRs were upregulated in the susceptible chicken breed. On postmortem examination, the resistant birds showed small lesions in the liver compared to large necrotic lesions in susceptible birds. The pathological manifestations and RNA sequencing results suggest a balancing link between resistance and infection tolerance in Kashmir faverolla. Here we also developed an online Poultry Infection Database (https://skuastk.org/pif/index.html), the first publicly available gene expression resource for disease resistance in chickens. The available database not only shows the data for gene expression in chicken tissues but also provides quick search, visualization and download capacity.

The aim of this study was to determine the antioxidant and antibacterial property of Nelumbo nucifers (lotus) leaf extract. Total phenolic content and DPPH (2, 2-diphenyl-1-picrylhydrozyl) scavenging methods were used to evaluate the antioxidant property of crude extract. DPPH scavenging capacity of extract varied significantly (p

Salmonella enterica serovar Typhimurium (S. Typhimurium) is a primary avian pathogen responsible for severe intestinal pathology in younger chickens and economic losses to poultry industry. Furthermore, S. Typhimurium is also able to cause infection in humans, characterized by acute gastrointestinal disease. A study was conducted to investigate antibody response and expression kinetics of interferon gamma (IFNγ), interleukin (IL-12, and IL-18) genes in broiler chicken at 0, 1, 3, 5, 7, 9, 11, 13, and 15 D post infection following experimental infection of S. Typhimurium. Immunological studies showed higher titres of IgG and IgM in the infected group as compared to the age-matched un-infected control group. The Real-Time PCR-based gene expression analysis revealed significant increase of IFNγ, IL-12, and IL-18 mRNA levels in the infected group as compared to their respective controls (P < 0.05). The present study shall help in understanding the immune responses in birds, thus allowing development of more effective vaccines and vaccination strategies.

The study was aimed to evaluate and compare the healing potential of mesenchymal stem cells (MSCs) derived from two common sources (iliac crest derived bone marrow and omental fat) in a full thickness skin wound model. Bone marrow derived MSCs clinical efficacy in the repair of cattle teat fistulae (cutaneous and muco-cutaneous wounds) was also evaluated. In a completely randomized placebo controlled experimental full thickness skin wound model, n=36 were randomly divided into three equal groups: groups I, II and III receiving Phosphate buffered saline (PBS), BM-MSCs and adipose tissue MSCs (AD-MSCs), respectively. Grossly early reduction in inflammation and enhanced epithelialization in the cell-treated groups as compared to the control was seen. Microscopy, ultramicroscopy, gene expression analysis and mechanical testing revealed better and early matrix formation with a reduced scar formation and a higher tensile strength in the cell-treated groups as compared to the control. An overall comparable healing in the cell treated groups was observed, although BM-MSCs had led to the better matrix formation tending to scarless healing while the AD-MSCs had led to the early wound closure with a good tissue strength. In the case controlled bovine clinical teat injuries study (n=17) repaired surgically, BM-MSCs (n=13) or PBS (n=4) was injected locally. In surgico-MSCs treated cases, 84.6% non-recurrence rate was observed as compared to the 50% seen in the control. It was concluded that MSCs irrespective of the donor tissue have potential to improve healing of full thickness cutaneous wounds and/ fistulae.

Mastitis or inflammation of the mammary gland is a highly economic and deadly alarming disease for the dairy sector as well as policymakers caused by microbial infection. Transcriptomic and proteomic approaches have been widely employed to identify the underlying molecular mechanisms of bacterial infections in the mammary gland. Numerous differentially expressed mRNAs, miRNAs, and proteins together with their associated signaling pathways have been identified during bacterial infection, paving the way for analysis of their biological functions. Long noncoding RNAs (lncRNAs) are important regulators of multiple biological processes. However, little is known regarding their role in bacterial infection in mammary epithelial cells. Hence, RNA-sequencing was performed by infecting primary mammary epithelial cells (pMECs) with both gram-negative (E. coli) and gram-positive bacteria (S. aureus). Using stringent pipeline, a set of 1957 known and 1175 novel lncRNAs were identified, among which, 112 lncRNAs were found differentially expressed in bacteria challenged PMECs compared with the control. Additionally, potential targets of the lncRNAs were predicted in cis- and trans-configuration. KEGG analysis revealed that DE lncRNAs were associated with at least 15 immune-related pathways. Therefore, our study revealed that bacterial challenge triggers the expression of lncRNAs associated with immune response and defense mechanisms in goat mammary epithelial cells.

The current study demonstrates the culture characteristics of adipose tissue and bone marrow-derived mesenchymal stem cells (MSC). The study evaluates the effect of ambient temperature, physiological status of the donor and the tissue source on sheep (Ovis aries) mesenchymal stem cells. The tissue samples were harvested from full term pregnant female sheep (n = 9) and male sheep (n = 10). Adipose tissue was harvested from n = 9 sheep and bone marrow from n = 10 sheep. The samples (adipose tissue, n = 2, bone marrow, n = 3) transported at cold ambient temperature (&lt, 10 °C) failed to yield MSCs while those (n = 14) at higher (&gt, 20 °C) ambient temperature successfully yielded MSCs. Bone marrow mononuclear cell (MNC) fraction was higher than the adipose tissue-derived stromal vascular fraction (SVF), but the percent adherent cells (PAC) was higher in the later cell fraction. Adipose tissue-derived MSCs from the full term female sheep had a significantly (p &lt, 0.05) higher proliferation potential as compared to those of the male sheep-derived MSCs. Female sheep MSCs also had rapid differentiation potential. The cryopreserved MSCs had morphological features comparable to that of the fresh cells. In conclusion, the tissue type and physiological status of donor animal may affect MSCs’ characteristics and should be taken into consideration while applying in clinical settings.

In healthcare system, the predictive modelling procedure for risk estimation of cardiovascular disease is extremely challenging and an inevitable task. Therefore, the attempt to clinically examine medical databases through conventional and leading-edge machine learning technologies is contemplated to be valuable, accurate and more importantly economical substitute for medical practitioners. In this research study, primarily we have exploited both individual learning algorithms and ensemble approaches including BayesNet, J48, KNN, multilayer perceptron, Naive Bayes, random tree and random forest for prediction purposes. After analysing the performance of these classifiers, J48 attained noteworthy accuracy of 70.77% than other classifiers. We then employed new fangled techniques comprising TENSORFLOW, PYTORCH and KERAS on the same dataset acquired from Stanford online repository. The empirical results demonstrate that KERAS achieved an outstanding prediction accuracy of 80% in contrast to entire set of machine learning algorithms which were taken under investigation. Furthermore, based on the performance improvisation in prediction accuracy of cardiovascular disease, a novel prediction model was propounded after conducting performance analysis on both approaches (conventional and cutting-edge technologies). The principle objective behind this research study was the pursuit for fitting approaches that can lead to better prediction accuracy and reliability of diagnostic performance of cardiovascular disease.

Characteristic features like self-renewal, multilineage differentiation potential, and immune-modulatory/anti-inflammatory properties, besides the ability to mobilize and home distant tissues make stem cells (SCs) a lifeline for an individual. Stem cells (SCs) if could be harvested and expanded without any abnormal change may be utilized as an all-in-one solution to numerous clinical ailments. However, slender understanding of their basic physiological properties, including expression potential, behavioral alternations during culture, and the effect of niche/microenvironment has currently restricted the clinical application of SCs. Among various types of SCs, mesenchymal stem cells (MSCs) are extensively studied due to their easy availability, straightforward harvesting, and culturing procedures, besides, their less likelihood to produce teratogens. Large ruminant MSCs have been harvested from various adult tissues and fetal membranes and are well characterized under in vitro conditions but unlike human or other domestic animals in vivo studies on cattle/buffalo MSCs have mostly been aimed at improving the animals' production potential. In this document, we focused on the status and potential application of MSCs in cattle and buffalo.

Conventional somatic cell nuclear transfer (SCNT) technique of in vitro production of cloned embryos involves use of costly and complicated micromanipulators. Handmade cloning (HMC) technique has been applied as efficient and cost-effective alternative in many livestock species. The aim of the present study was to compare the efficiency of in vitro production and in vitro development of cloned sheep embryos by the two techniques. Cloned embryos were produced by conventional SCNT using micromanipulator apparatus and by HMC technique. Enucleation efficiency and efficiency of fusion with somatic cell (nucleus donor) were compared. Cleavage percentage was observed on day 2 of in vitro culture (IVC), and morula and blastocyst percentages were calculated on day 7 of IVC. Higher enucleation efficiency (96.98 ± 1.01 vs. 93.62 ± 1.03; p .05) as well as fusion efficiency was obtained with HMC technique than with conventional SCNT (96.26 ± 1.34 vs. 92.63 ± 0.70, p .05); 181 cloned sheep embryos were produced in vitro by conventional SCNT and 92 by HMC. Cleavage percentage observed on day 2 of in vitro culture was higher in HMC than SCNT (66.92 ± 3.72 vs. 55.97 ± 2.5, respectively, p .05). Morula percentage obtained was higher in SCNT than HMC (44.12 ± 2.93 vs. 30.43 ± 6.79, respectively, p .05), whereas blastocyst percentage obtained by HMC was higher (12.46 ± 4.96) than SCNT (5.31 ± 2.25; p .05). It was inferred that HMC technique provides a cost-effective and efficient method of in vitro production of cloned sheep embryos with a comparatively simpler technique with a possibility of automation. Efficiency of cloned embryo production could be improved further by propagating and standardizing this technique.

The present study was designed to test the effect of different levels of idebenone, a potent antioxidant on the quality of ram semen at post thaw. Eighteen (18) ejaculates were collected and extended with tris extender supplemented with no antioxidant (CON), with 2 μM idebenone (Id2), 5 μM idebenone (Id5), 7.5 μM idebenone (Id7.5) and 10 μM idebenone (Id10). The sperm quality was determined in terms of percent sperm motility, live sperm percentage, percent hypoosmotic swelling test (HOST) positive spermatozoa and percent intact acrosome (PIA). Moreover, malondialdehyde (MDA) level, an end product of lipid peroxidation (LPO) was also measured at post thaw both in seminal plasma and sperm cell. At post thaw, the percent sperm motility was significantly higher (p 0.05) for Id10 as compared to Id2, Id5, Id7.5 and control. The live sperm percentage was non-significantly (p 0.05) higher for Id10 as compared to control, Id5 and Id7.5 but significantly higher than Id2. The percent HOST positive spermatozoa was significantly higher (p 0.05) for Id10 than control, Id2 and Id5. The MDA level in seminal plasma was significantly lower (p 0.05) for Id10 than control and Id2. The MDA level in spermatozoa did show similar trend as in seminal plasma. Further, all the sperm parameters at all idebenone levels declined significantly from pre freeze to post thaw. In conclusion, idebenone at 10 μM level improved post thaw sperm quality by mitigating peroxidative stress, hence could be considered as a promising antioxidant additive for cryopreservation of ram semen.

This article released online on March 5, 2019 as advance publication was withdrawn from consideration for publication in Journal of Reproduction and Development at author's request.

The present study aimed to establish a zona free (handmade cloning) embryo production system for Pashmina goat embryos. Abattoir derived oocytes were matured in vitro; after maturation, oocytes were enucleated and fused with somatic cells derived from an adult Pashmina goat tissue. The reconstructs were activated using a calcium ionophore-DMAP procedure. The embryos were distributed into two experimental groups. In Experiment 1, the embryos were cultured in one of the following four culture media (i) G1.G2 media (ii) Modified synthetic oviduct fluid (mSOF) (iii) Research vitro cleave media (RVCL) and (iv) Embryo development media (EDM), and were cultured for 7 days. The cleavage rates in G1.G2, RVCL, and mSOF were higher than those in EDM (86.8, 82.4, 77.3, and 68.8%, respectively). Blastocyst rates were higher in RVCL than those in mSOF, EDM, and G1.G2 (15.0, 10.5, 4.9, and 2.2%, respectively). In experiment 2, the embryos were cultured in five different culture systems: (i) Flat surface (FS), (ii) Well in drop (WID), (iii) Well of well (WOW), (iv) Micro drop, and (v) Hanging drop, for 7 days. The cleavage rates in FS and WID were higher than those in WOW, Micro drop, and Hanging drop (84.3, 81.2, 73.6, 73.5, and 70.3%, respectively). The blastocyst rates were higher in WID than those in WOW, Micro drop, Hanging drop, and FS systems (21.6, 13.7, 11.5, 10.9, and 3.9%, respectively). The embryos produced in experiment 2 were transferred to synchronized recipients. Of the three pregnancies established on day 40, one resulted in the delivery of a healthy Pashmina kid.

A blinded, randomized, prospective experimental study was planned to compare the use of 1% and 2% lidocaine hydrochloride (LH) for distal paravertebral nerve block (DPNB) in Bakerwal goats undergoing reproductive laparoscopy. The goats (n = 13) were randomly allotted to two treatments: Treatment I (n = 7) and Treatment II (n = 6) and injected 15.0 mL of either 1% or 2% LH. Nociceptive stimuli during pre- and postoperative periods comprised of superficial and deep pinpricks. The reaction of the animals to the surgical intervention and alterations in their behaviour were recorded intraoperatively. The time to inject the anesthetic (5.4 ± 1.6 and 4.9 ± 0.9 min), onset time (2.6 ± 1.6 and 2.3 ± 1.4 min), the incision length (5.0 ± 1.4 cm and 4.7 ± 1.7 cm) and the sensory blockade duration (121 ± 18 and 123 ± 17 min) in Treatment I and Treatment II goats respectively showed no significant difference. In Treatment I goats the anesthetic dose (5.2 ± 1.1 mg kg -1) injected was significantly less than Treatment II (9.8 ± 0.8 mg kg -1) but their laparoscopy time (73 ± 25 min) was significantly more than Treatment II (56 ± 4 min) animals. The satisfactory sensory blockade lasted for similar period; Treatment I (121 ± 18 min) and Treatment II (123 ± 17 min). The physiological parameters fluctuated most of the times within the normal range. Signs of toxicity, acute pain or distress were not recorded in any of the animals. From this study it is concluded that satisfactory sensory blockade develops after DPNB using either 2% or 1% LH in goats. However, further trials involving higher number of healthy and ailing animals and evaluation of other nerve blocks are needed before advocating the routine use of 1% preparation in the caprine clinical practice.

Jersey and Kashmiri cattle are important dairy breeds that contribute significantly to the total milk production of the Indian northern state of Jammu and Kashmir. The Kashmiri cattle germplasm has been extensively diluted through crossbreeding with Jersey cattle with the goal of enhancing its milk production ability. However, crossbred animals are prone to diseases resulting to unsustainable milk production. This study aimed to provide a comprehensive transcriptome profile of mammary gland epithelial cells at different stages of lactation and to find key differences in genes and pathways regulating milk traits between Jersey and Kashmiri cattle. Mammary epithelial cells (MEC) isolated from milk obtained from six lactating cows (three Jersey and three Kashmiri cattle) on day 15 (D15), D90 and D250 in milk, representing early, mid and late lactation, respectively were used. RNA isolated from MEC was subjected to next-generation RNA sequencing and bioinformatics processing. Casein and whey protein genes were found to be highly expressed throughout the lactation stages in both breeds. Largest differences in differentially expressed genes (DEG) were between D15 vs D90 (1,805 genes) in Kashmiri cattle and, D15 vs D250 (3,392 genes) in Jersey cattle. A total of 1,103, 1,356 and 1,397 genes were differentially expressed between Kashmiri and Jersey cattle on D15, D90 and D250, respectively. Antioxidant genes like RPLPO and RPS28 were highly expressed in Kashmiri cattle. Differentially expressed genes in both Kashmiri and Jersey were enriched for multicellular organismal process, receptor activity, catalytic activity, signal transducer activity, macromolecular complex and developmental process gene ontology terms. Whereas, biological regulation, endopeptidase activity and response to stimulus were enriched in Kashmiri cattle and, reproduction and immune system process were enriched in Jersey cattle. Most of the pathways responsible for regulation of milk production like JAK-STAT, p38 MAPK pathway, PI3 kinase pathway were enriched by DEG in Jersey cattle only. Although Kashmiri has poor milk production efficiency, the present study suggests possible physicochemical and antioxidant properties of Kashmiri cattle milk that needs to be further explored.

The present study aimed to establish a zona free (handmade cloning) embryo production system for Pashmina goat embryos. Abattoir derived oocytes were matured in vitro; after maturation, oocytes were enucleated and fused with somatic cells derived from an adult Pashmina goat tissue. The reconstructs were activated using a calcium ionophore-DMAP procedure. The embryos were distributed into two experimental groups. In Experiment 1, the embryos were cultured in one of the following four culture media (i) G1.G2 media (ii) Modified synthetic oviduct fluid (mSOF) (iii) Research vitro cleave media (RVCL) and (iv) Embryo development media (EDM), and were cultured for 7 days. The cleavage rates in G1.G2, RVCL, and mSOF were higher than those in EDM (86.8, 82.4, 77.3, and 68.8%, respectively). Blastocyst rates were higher in RVCL than those in mSOF, EDM, and G1.G2 (15.0, 10.5, 4.9, and 2.2%, respectively). In experiment 2, the embryos were cultured in five different culture systems: (i) Flat surface (FS), (ii) Well in drop (WID), (iii) Well of well (WOW), (iv) Micro drop, and (v) Hanging drop, for 7 days. The cleavage rates in FS and WID were higher than those in WOW, Micro drop, and Hanging drop (84.3, 81.2, 73.6, 73.5, and 70.3%, respectively). The blastocyst rates were higher in WID than those in WOW, Micro drop, Hanging drop, and FS systems (21.6, 13.7, 11.5, 10.9, and 3.9%, respectively). The embryos produced in experiment 2 were transferred to synchronized recipients. Of the three pregnancies established on day 40, one resulted in the delivery of a healthy Pashmina kid.

The Bakerwal breed of goat in Kashmir valley is a good meat purpose breed of goat. That attains an appreciable body weight under a low input production system. As these goats are mainly reared by Gujjar and Bakarwal tribes of the J & K state, so they usually are fed with the weeds, herbs, shrubs and grasses of pastures that will otherwise go waste. These goats constitute the main livestock wealth. With the good productive and reproductive potential, it makes these animals an important animal protein source for developing countries like India. The myostatin gene (GDF8) is important in the physiology of stock animals because its product produces a direct effect on muscle development and consequently also on meat production. The myostatin sequence is known in several mammalian species and shows a high degree of amino acid sequence conservation, although several alterations in the intron and exon regions have been identified. The objective of our work is to characterize the myostatin coding regions using gene sequencing and polymerase chain reaction methods of Capra hircus (Bakerwal breed) and to compare them with the Ovis aries and other livestock species of animal, looking for variations in nucleotide and protein sequences. As mutations in the myostatin gene can inactivate its expression and result in a non-functional protein, which leads to increase in muscle growth in many species. In this way, we are able to identify 3 alterations on the presumed myostatin protein sequence as compared to non double-muscled ovine sequences.

Pashmina goat inhabits the high altitude cold arid desert of Ladakh, India. This goat is known for its finest and costliest under fiber. Though the under fiber may be a part of its complex thermoregulation mechanism, the genetics of its adaptability under cold conditions is not known. As an attempt to understand its adaptive genetics, and the role of RNA-binding proteins at the cellular response, this study was conducted to characterize the RBM3 gene in Pashmina goat and its expression during hypothermia. The ORF of Pashmina RBM3 gene was 273 bp. Phylogenetic analysis revealed that Pashmina RBM3 is closely related to Bos taurus RBM3. Pashmina RBM3 was characterized by comparative modeling studies. The final 3-D model contained two α-helices and four β-sheets. qRT-PCR data showed that Pashmina RBM3 gene expression was significantly higher (P

Dystrophin protein in association with several other cellular proteins and glycoproteins leads to the formation of a large multifaceted protein complex at the cell membrane referred to as dystrophin glycoprotein complex (DGC), that serves distinct functions in cell signaling and maintaining the membrane stability as well as integrity. In accordance with this, several findings suggest exquisite role of DGC in signaling pathways associated with cell development and/or maintenance of homeostasis. In the present review, we summarize the established facts about the various components of this complex with emphasis on recent insights into specific contribution of the DGC in cell signaling at the membrane. We have also discussed the recent advances made in exploring the molecular associations of DGC components within the cells and the functional implications of these interactions. Our review would help to comprehend the composition, role, and functioning of DGC and may lead to a deeper understanding of its role in several human diseases.

Immune responses combat various infectious agents by inducing inflammatory responses, antimicrobial pathways and adaptive immunity. The polygenic responses to these external stimuli are temporally and coordinately regulated. Specific lncRNAs are induced to modulate innate and adaptive immune responses which can function through various target interactions like RNA-DNA, RNA-RNA, and RNA-protein interaction and hence affect the immunogenic regulation at various stages of gene expression. LncRNA are found to be present in various immune cells like monocytes, macrophages, dendritic cells, neutrophils, T cells and B cells. They have been shown to be involved in many biological processes, including the regulation of the expression of genes, the dosage compensation and genomics imprinting, but the knowledge how lncRNAs are regulated and how they alter cell differentiation/function is still obscure. Further dysregulation of lncRNA has been seen in many diseases, but as yet very less research has been carried out to understand the role of lncRNAs in regulation during host-pathogens interactions. In this review, we summarize the functional developments and mechanism of action of lncRNAs, in immunity and defense of host against pathogens.

Mesenchymal stem cells (MSCs) being available in numerous adult tissues and foetal membranes have been harvested from almost all these sources in dog. Due to the presence of miniscule number of MSCs in these tissues, culture expansion followed by their characterization becomes imperative. These characterized cells have been made subject in preclinical experimental and clinical settings in dogs. The studies have variably but undeniably supported their therapeutic effects, although currently inconclusive. Number of dog clinical ailments compeer to that of human. Therapeutic benefits of dMSCs in these ailments may provide proof-of-therapeutic principle for human medicine. The current chapter details dMSCs culture characteristics and potential applications.

Goats (n=12) undergoing laparoscopy assisted embryo transfer were randomly allotted to two groups (I and II) and injected same volume of ropivacaine hydrochloride at 1.0 mg/kg and 0.5 mg/kg body weight, respectively, at the lumbosacral epidural space. The hind quarters of all the animals were lifted up for the first 3.0 minutes following injection. Immediately after induction the animals were restrained in dorsal recumbency in Trendelenburg position in a cradle. Laparoscopy was performed after achieving pneumoperitoneum using filtered room air. Regional analgesia and changes in physiological parameters were recorded. The mean induction time in animals of group I (n=6) was 12.666 ± 1.994 minutes. In these animals the analgesia extended up to the umbilical region and lasted for 60 minutes. Only two animals in group II were satisfactorily induced in 11.333 ± 2.333 minutes. In animals of group I, the time taken for regaining the full motor power was significantly long (405 ± 46.314 min) when compared to group II goats (95 ± 9.219 min). From this study it was concluded that ropivacaine did not produce adequate analgesia in most of the goats at 0.5 mg/kg. When used at 1.0 mg/kg, it produced satisfactory regional analgesia lasting for one hour but the prolonged motor loss precludes its use. Additional studies using ropivacaine hydrochloride at doses in between the two extremes used here may be undertaken before recommending it for lumbosacral anaesthesia in goats undergoing laparoscopy.

The gradual decline in the genetic diversity of farm animals has threatened their survival and risk of their extinction has increased many fold in the recent past. Endangered species could be rescued using interspecies embryo production. The objective of this study was to investigate the effect of three different culture media on the development of Handmade cloned intraspecies (goat-goat) and interspecies (goat-sheep) embryo reconstructs. Research vitro cleave media (RVCL) yielded higher cleavage and morula-blastocyst development in intraspecies and interspecies nuclear transfer groups compared with G1.G2 and modified synthetic oviductal fluid (mSOFaaci). Cleavage frequency of intraspecies cloned embryos in RVCL, mSOFaaci, and G1.G2 did not differ significantly (87.12%, 82.45%, and 92.52%, respectively). However, the morula/blastocyst frequency in RVCL was greater in mSOFaaci and G1.G2 (51.18% vs. 38.28% vs. 36.50%, respectively). Cleavage and morula/blastocyst frequency in interspecies cloned embryos was greater in RVCL than in mSOFaaci and G1.G2 (76.14% and 42.3% vs. 65.9% and 38.3% vs. 58.56% and 33.1%, respectively). Goat oocytes were parthenogenetically activated and cultured in RVCL, mSOFaaci, and G1.G2 and kept as control. Cleavage and morula/blastocyst frequency in this group was greater in RVCL than in mSOFaaci and G1.G2 (89.66% and 65.26% vs. 85.44% and 48.05% vs. 86.58% and 42.06%, respectively). Conclusively, the results suggest that not only can the interspecies embryos of goat be produced using sheep oocytes as donor cytoplast but also the percentages can be improved by using RVCL media for culturing of the embryos.

The aim of this study was to compare the survival and in vitro development of sheep oocytes after open pulled straw vitrification and different final concentrations of permeable cryoprotectants. In 5 identical replicates of two experiments, in vitro matured (IVM) oocytes were vitrified by the Open Pulled Straw (OPS) method, then warmed, and the surviving ones were subjected to parthenogenetic activation. In Experiment 1, survival rate of oocytes after vitrification in 33% ethylene glycol was higher than in 33% DMSO or a mixture of 17.5% ethylene glycol and 17.5% DMSO (87.64 vs. 77.43 vs. 69.39%, respectively). The cleavage and blastocyst rates were higher after vitrification in mixture group than in ethylene glycol and DMSO (46.81 and 15.5 vs. 37.55 and 9.12 vs. 29.51 and 6.40%, for cleavage and blastocyst rates in different groups, respectively). In Experiment 2, elevated concentrations of vitrification solutions were used. The survival rate was higher after vitrification in 40% ethylene glycol and in the mixture of 20% ethylene glycol and 20% DMSO than in 40% DMSO (90.22 vs. 87.56 vs. 75.34%, respectively). Cleavage and blastocyst rates were also higher in the ethylene glycol and ethylene glycol – DMSO mixture group than in DMSO alone group (50.67 and17.60 vs. 49.13 and 14.45 vs. 33.86 and 9.81% for cleavage and blastocyst rates in different groups, respectively). The survival rates between the two experimental groups was higher in 40% ethylene glycol group, 40% mixture group and 33% ethylene glycol group than in 40% DMSO group, 33% mixture group and 33% DMSO group. Cleavage and blastocyst rates were higher in 40% ethylene glycol group, 40% mixture group and 33% mixture group than in 40% DMSO group, 33% ethylene glycol group and 33% DMSO group. All cleavage and blastocyst rates in both the experiments were lower than those of the non-vitrified control group (87.00 and 45.00, respectively). In conclusion, although ethylene glycol group and ethylene glycol – DMSO mixture group gave better survival and cleavage – blastocyst rates than DMSO group, the survival rates were lower than the control group and hence the technique could be further improved to get better results after OPS vitrification of IVM sheep oocytes.

Common bean ( Phaseolus vulgaris L.) is a legume of appreciable importance and usefulness worldwide to the human population providing food and feed. It is rich in high-quality protein, energy, fiber and micronutrients especially iron, zinc, and pro-vitamin A; and possesses potentially disease-preventing and health-promoting compounds. The recently published genome sequence of common bean is an important landmark in common bean research, opening new avenues for understanding its genetics in depth. This legume crop is affected by diverse biotic and abiotic stresses severely limiting its productivity. Looking at the trend of increasing world population and the need for food crops best suited to the health of humankind, the legumes will be in great demand, including the common bean mostly for its nutritive values. Hence the need for new research in understanding the biology of this crop brings us to utilize and apply high-throughput omics approaches. In this mini-review our focus will be on the need for proteomics studies in common bean, potential of proteomics for understanding genetic regulation under abiotic and biotic stresses and how proteogenomics will lead to nutritional improvement. We will also discuss future proteomics-based strategies that must be adopted to mine new genomic resources by identifying molecular switches regulating various biological processes. Significance Common bean is regarded as “grain of hope” for the poor, being rich in high-quality protein, energy, fiber and micronutrients (iron, zinc, pro-vitamin A); and possesses potentially disease-preventing and health-promoting compounds. Increasing world population and the need for food crops best suited to the health of humankind, puts legumes into great demand, which includes the common bean mostly. An important landmark in common bean research was the recent publication of its genome sequence, opening new avenues for understanding its genetics in depth. This legume crop is affected by diverse biotic and abiotic stresses severely limiting its productivity. Therefore, the need for new research in understanding the biology of this crop brings us to utilize and apply high-throughput omics approaches. Proteomics can be used to track all the candidate proteins/genes responsible for a biological process under specific conditions in a particular tissue. The potential of proteomics will not only help in determining the functions of a large number of genes in a single experiment but will also be a useful tool to mine new genes that can provide solution to various problems (abiotic stress, biotic stress, nutritional improvement, etc). We believe that a combined approach including breeding along with omics tools will lead towards attaining sustainability in legumes, including common bean.

Contents When buffalo embryonic stem (ES) cell–like cells that expressed surface markers SSEA-4, TRA-1-60, TRA-1-81, CD9 and CD90 and intracellular markers OCT4, SOX2 and FOXD3, as shown by immunofluorescence, and that expressed REX-1 and NUCLEOSTEMIN as confirmed by RT-PCR, were subjected to suspension culture in hanging drops in absence of LIF and buffalo foetal fibroblast feeder layer support, they differentiated to form three-dimensional embryoid bodies (EBs). Of 231 EBs examined on Day 3 of suspension culture, 141 (61.3 ± 3.09%) were of compact type, whereas 90 (38.4 ± 3.12%) were of cystic type. The cells obtained from EBs were found to express NF-68 and NESTIN (ectodermal lineage), BMP-4 and α-skeletal actin (mesodermal lineage), and α-fetoprotein, GATA-4 and HNF-4 (endodermal lineage). When these EBs were cultured on gelatin-coated dishes, they spontaneously differentiated to several cell types such as epithelial- and neuron-like cells. When EBs were cultured in the presence of 1 or 2% DMSO or 10−8 m or 10−7 m retinoic acid for 25 days, ES cells could be directed to form muscle cell–like cells, the identity of which was confirmed by expression of α-actinin by immunofluorescence and of MYF-5, MYOD and MYOGENIN genes by RT-PCR. MYOD was first detected on Day 10 in both treatment groups and on Day 15 in controls, whereas MYOGENIN was first detected on Day 10, Day 15 and Day 25 in the presence of retinoic acid, in the presence of DMSO and in controls, respectively. The present study demonstrates the ability of buffalo ES cell–like cells to undergo directed differentiation to cells of skeletal myogenic lineage.

The aim of the present study is to compare the ability of homologous and heterologous embryonic fibroblast feeder layers to support isolation and proliferation of buffalo ES-like cells generated from hatched and expanded blastocysts produced by in vitro fertilization and characterization of derived cells through expression of pluripotent markers.Embryonic stem cells were derived from hatched and expanded blastocysts through intact blastocyst culture and enzymatic method respectively and compared for proliferation rate on homologous (buffalo) and heterologous feeder layers (goat and sheep).A total of 69 hatched and 83 expanded blastocysts were used for isolation of inner cell masses which were seeded on buffalo, goat and sheep embryonic feeder layers. Following seeding, attachment rate, primary colony formation rate and survival to maximum number of passages were observed to be higher on homologous feeder layers.Upon comparison of different feeder layer cells for derivation and maintenance of buffalo ES-like cells from hatched and expanded blastocysts, buffalo embryonic fibroblast cells were able to provide a better environment for maintaining pluripotency in culture conditions.

The possibility of producing interspecies handmade cloned (iHMC) embryos by nuclear transfer from donor cells of cattle, goat and rat using buffalo oocytes as recipient cytoplasts was explored. Zona-free buffalo oocytes were enucleated by protrusion cone-guided bisection with a microblade. After electrofusion with somatic cells, reconstructed oocytes were activated by calcimycin A23187, treated with 6-dimethylaminopurine and were cultured in K-RVCL-50® medium for 8 days. Although the cleavage rate was not significantly different when buffalo, cattle, goat or rat cells were used as donor nuclei (74.6 ± 3.8, 82.8 ± 5.3, 86.0 ± 4.9 and 82.3 ± 3.6%, respectively), the blastocyst rate was significantly higher (P

The aim of this study was to determine the antioxidant and antibacterial property of Nelumbo nucifers (lotus) leaf extract. Total phenolic content and DPPH (2, 2-diphenyl-1-picrylhydrozyl) scavenging methods were used to evaluate the antioxidant property of crude extract. DPPH scavenging capacity of extract varied significantly (p

In this study, inner cell mass (ICM) cells were isolated from in vitro produced buffalo blastocysts and were cultured on mitomycin-C treated buffalo foetal fibroblast feeder layer for producing embryonic stem (ES) cells. Among different sources (hatched vs expanded blastocysts) or methods (enzymatic vs mechanical), mechanical isolation of ICM from hatched blastocysts resulted in the highest primary colony formation rate and the maximum passage number up to which ES cells survived. Putative ES cells expressed alkaline phosphatase and exhibited a normal karyotype up to passage 7. Putative ES cells and embryos at 2- to 4-cell, 8- to 16-cell, morula and blastocyst stages strongly expressed stage-specific embryonic antigen (SSEA)-4 but lacked expressions of SSEA-1 and SSEA-3. Putative ES cells also expressed tumour rejection antigen (TRA)-1-60, TRA-1-81 and Oct4. Whereas in all early embryonic stages, TRA-1-60 was observed only in the periplasmic space, and TRA-1-81 expression was observed as small spots at a few places inside the embryos, both these markers were expressed by ICM. Oct4 expression, which was observed at all the embryonic stages and also in the trophectoderm, was the strongest in the ICM. Buffalo putative ES cells possess a unique pluripotency-related surface antigen phenotype, which resembles that of the ICM.