Effect of cryopreservation and capacitation on expression patterns of ATP synthesis associated genes in bubaline spermatozoa.

This study investigated the expression patterns of certain ATP synthesis associated genes (GAPDHS, PGK2, ENO4 and MDH2) in fresh and frozen-thawed bubaline spermatozoa vis-à-vis capacitation. Eight adult Murrah buffalo bulls, aged 5-7 years, were randomly selected from a bull farm and fresh semen was collected once a week for four weeks (n = 32) from the selected bulls. Simultaneously, frozen straws of the same bulls were collected (n = 32, four straws were pooled per sample).

Carbohydrate-Small Molecule Hybrids as Lead Compounds Targeting IL-6 Signaling.

In the past 25 years, a number of efforts have been made toward the development of small molecule interleukin-6 (IL-6) signaling inhibitors, but none have been approved to date. Monosaccharides are a diverse class of bioactive compounds, but thus far have been unexplored as a scaffold for small molecule IL-6-signaling inhibitor design. Therefore, in this present communication, we combined a structure-based drug design approach with carbohydrate building blocks to design and synthesize novel IL-6-signaling inhibitors targeting glycoprotein 130 (gp130).

Perspectives of pluripotent stem cells in livestock.

The recent progress in derivation of pluripotent stem cells (PSCs) from farm animals opens new approaches not only for reproduction, genetic engineering, treatment and conservation of these species, but also for screening novel drugs for their efficacy and toxicity, and modelling of human diseases. Initial attempts to derive PSCs from the inner cell mass of blastocyst stages in farm animals were largely unsuccessful as either the cells survived for only a few passages, or lost their cellular potency; indicating that the protocols which allowed the derivation of murine or human embryonic stem (ES) cells were not sufficient to support the maintenance of ES cells from farm animals. This scenario changed by the innovation of induced pluripotency and by the development of the 3 inhibitor culture conditions to support naïve pluripotency in ES cells from livestock species.

Transcriptional and translational abundance of visfatin (NAMPT) in buffalo ovary during estrous cycle and its in vitro effect on steroidogenesis.

Visfatin is a highly conserved adipokine protein having multiple biological effects, including regulation of reproduction. Evidence in recent years has shown a pivotal role of visfatin in ovarian functions. The present study was conducted to evaluate the mRNA and protein abundance of visfatin in ovarian follicles and corpora lutea (CL) during different stages of their development in the ovary of water buffalo (Bubalus bubalis) and to investigate the role of visfatin on estradiol (E) and progesterone (P) secretion.

Applications, challenges, and strategies in the use of nanoparticles as feed additives in equine nutrition.

The rapid expansion of nanotechnology has been transforming the food industry by increasing market share and expenditure. Although nanotechnology offers promising benefits as feed additives, their usage in equines is primarily geared toward immunotherapy, hyper-immunization techniques, drug delivery systems, grooming activities, and therapeutic purposes. Nanoparticles could be engaged as alternatives for antibiotic feed additives to prevent foal diarrhea.

A versatile bulk electrotransfection protocol for murine embryonic fibroblasts and iPS cells.

Although electroporation has been widely accepted as the main gene transfer tool, there is still considerable scope to improve the electroporation efficiency of exogenous DNAs into primary cells. Here, we developed a square-wave pulsing protocol using OptiMEM-GlutaMAX for highly efficient transfection of murine embryonic fibroblasts (MEF) and induced pluripotency stem (iPS) cells using reporter genes as well as gRNA/Cas9-encoding plasmids. An electrotransfection efficiency of > 95% was achieved for both MEF and iPS cells using reporter-encoding plasmids.

Systematic optimization of square-wave electroporation conditions for bovine primary fibroblasts.

Background: Gene transfer by electroporation is an established method for the non-viral mediated transfection of mammalian cells. Primary cells pose a particular challenge for electroporation-mediated gene transfer, since they are more vulnerable than immortalized cells, and have a limited proliferative capacity. Improving the gene transfer by using square wave electroporation in difficult to transfect cells, like bovine fetal fibroblasts, is a prerequisite for transgenic and further downstream experiments.

Environmental sustainability assessment of tropical dairy buffalo farming vis-a-vis sustainable feed replacement strategy.

Feeding management in dairy animals is crucial for ensuring optimal production apart from making the farming as a whole, a more sustainable activity. In our study we evaluated the production and environmental effects of two different feeding regimens i.e.

Assessment of eco-sustainability vis-à-vis zoo-technical attributes of soybean meal (SBM) replacement with varying levels of coated urea in Nellore sheep (Ovis aries).

The contemporary environmental-stewardship programmes primarily aimed at curbing the global warming potential by adopting a multidisciplinary approach. Manipulating the feeding strategies has great potential in reducing the environmental footprints of livestock production. This study intends to assess the effect of soybean meal (SBM) replacement with varying levels of coated urea (SRU) on both zoo-technical (nutrient digestibility, heat increment, and physio-biochemical parameters) and environmental attributes.

Implantation associated divergence in neutrophil glucocorticoid and cysteine-X-cysteine receptor genes in cattle.

The differential roles of Interferon-tau (IFNT) and Progesterone (P) in triggering pro- and anti-inflammatory responses during peri-implantation period were assessed in cows. An ex vivo study was done by isolating neutrophils from the animals post artificial insemination (AI) at defined intervals and analyzing the expression changes of Glucocorticoid receptor-α (GRα) and Interleukin-8 receptor (CXCR1) genes. An in vitro study was also done isolating neutrophils from prepubertal heifers subjecting them to IFNT and Progesterone P supplementation separately.

Exploring the concentration-dependent actions of interferon-τ on bovine neutrophils to understand the process of implantation.

Interferon-τ (IFNT) is a major cytokine produced by the ruminant trophoectoderm during the peri-implantation period exerting immunomodulatory actions on various cells including neutrophils. The current in vitro study was undertaken to analyze the concentration-dependent effects of IFNT on neutrophil gene dynamics to understand its possible role in implantation process. The neutrophils were isolated from the blood of heifers and were cultured subjecting them to different IFNT concentrations (1, 5, or 10 ng/mL).

Design and Synthesis of Novel 1,2,3-Triazole- and 2-Isoxazoline-Based Bis-Heterocycles as Immune Potentiators.

A series of novel bis-heterocycles encompassing isoxazolines and triazoles were synthesized through a novel one-pot procedure that involves in situ generation of nitrile oxide and its 1,3-dipolar cycloaddition to variedly substituted acrylates. The synthesized bis-heterocycles (8a-l) were subjected to in vitro lymphocyte proliferation assays followed by in vivo studies of the more active compounds (8g and 8h) to assess their influence on various aspects of the immune system like ex vivo splenocyte proliferation (T- and B-cell proliferation), antibody production (HA titer), delayed-type hypersensitivity reaction, T-cell subtypes (CD4 and CD8), cytokine production (IL-2, IFN-γ, and IL-4), NO (macrophage) production, and toxic effects. The results from the in vitro and in vivo studies establish that the tested compounds 8g and 8h possess excellent immunopotentiating activity.

Synthesis and Biological evaluation of novel 4β-[(5-substituted)-1,2,3,4-tetrazolyl] podophyllotoxins as anticancer compounds.

A series of novel 4β-[(5-substituted)-1,2,3,4-tetrazolyl] podophyllotoxin derivatives were synthesized by employing azide-nitrile click chemistry approach. All the derivatives were evaluated for their cytotoxicity against a panel of four human cancer cell lines and their IC50 values were found to be in the range of 2.4-29.

HSP70 as a marker of heat and humidity stress in Tarai buffalo.

Heat and humidity stress is a constant challenge to buffalo rearing under tropical climatic conditions. Heat shock proteins (HSPs) constitute a ubiquitous class of highly conserved proteins that contribute to cell survival during different conditions of stress. The present study was carried out in Tarai buffaloes to study the expression of HSP70 in their peripheral blood mononuclear cells during different seasons and establish it as a marker of heat and humidity stress in buffaloes.

Expression and localization of insulin-like growth factor system in corpus luteum during different stages of estrous cycle in water buffaloes (Bubalus bubalis) and the effect of insulin-like growth factor I on production of vascular endothelial growth factor and progesterone in luteal cells cultured in vitro.

This study investigated the expression and localization of insulin-like growth factor (IGF) system at different stages of buffalo CL and the role of IGF-I in stimulating vascular endothelial growth factor (VEGF) and progesterone (P4) production in cultured luteal cells. The mRNA expression of IGF system, VEGF, steroidogenic acute regulatory protein, P450scc, and hydroxysteroid dehydrogenase (HSD) was investigated by quantitative real-time polymerase chain reaction (PCR). Protein expression of IGF was demonstrated by Western blot and localization by immunohistochemistry.

Design and synthesis of novel 1,2,3-triazole derivatives of coronopilin as anti-cancer compounds.

A series of 1,2,3-triazole coronopilin congeners have been designed and synthesized by employing click chemistry approach starting from parthenin and evaluated for their cytotoxicity against a panel of six human cancer cell lines (PC-3, THP-1, HCT-15, HeLa, A-549 and MCF-7). While many compounds exhibited significant anticancer activity, compound 3a, was found to be the most promising analogue in this series with IC50 values of 3.1 μM on PC-3 cell line.

Expression and localization of ghrelin and its functional receptor in corpus luteum during different stages of estrous cycle and the modulatory role of ghrelin on progesterone production in cultured luteal cells in buffalo.

Evidence obtained during recent years provided has insight into the regulation of corpus luteum (CL) development, function, and regression by locally produced ghrelin. The present study was carried out to evaluate the expression and localization of ghrelin and its receptor (GHS-R1a) in bubaline CL during different stages of the estrous cycle and investigate the role of ghrelin on progesterone (P4) production along with messenger RNA (mRNA) expression of P4 synthesis intermediates. The mRNA and protein expression of ghrelin and GHS-R1a was significantly greater in mid- and late luteal phases.

Luteinizing hormone, insulin like growth factor-1, and epidermal growth factor stimulate vascular endothelial growth factor production in cultured bubaline granulosa cells.

The objective of this study was to characterize in vitro expression and secretion of vascular endothelial growth factor (VEGF) in bubaline granulosa cells (GC), grown in serum containing media supplemented with luteinizing hormone (LH), insulin like growth factor-1 (IGF-1), and epidermal growth factor (EGF) at three different doses and time durations. GCs were collected from ovarian follicles of varying diameters [Gp-I (small), 4-6 mm; Gp-II (medium), 7-9 mm; Gp-III (large), 10-13 mm; Gp-IV (pre-ovulatory), >13 mm]. In general, each of the three treatments resulted in a dose as well as time dependent increase in the mRNA expression and secretion of VEGF in the cultured GCs of Gp-IV follicles.

Expression and localization of locally produced growth factors regulating lymphangiogenesis during different stages of the estrous cycle in corpus luteum of buffalo (Bubalus bubalis).

Recent experiments using expression, immunolocalization, and cell culture approaches have provided leading insights into regulation of luteal angiogenesis by different growth factor systems and its role in the function of corpus luteum (CL) in buffalo. On the contrary, lymphangiogenesis and its regulation in the CL are still poorly understood. The aim of this study was to evaluate the expression and localization of lymphangiogenic factors (vascular endothelial growth factor [VEGF]-C and VEGFD), their receptor (VEGFR3), and lymphatic endothelial marker (LYVE1) in bubaline CL during different stages of the estrous cycle and to investigate functional role of VEGFC and VEGFD in luteal lymphangeogenesis.

Imaging features of a gelatin-thrombin matrix hemostatic agent in the intracranial surgical bed: a unique space-occupying pseudomass.

Background And Purpose: Absorbable gelatin-thrombin matrix is increasingly being used in neurosurgical procedures; unlike other hemostats, the stable matrix is left undisturbed and fills the surgical bed after achieving hemostasis. We investigated the immediate postoperative radiographic imaging appearance of the gelatin-thrombin matrix in intracranial operative beds.

Materials And Methods: Thirty-one consecutive patients (18 men, 13 women; mean age, 59 years) with 34 surgical cavities, had 31 brain MRIs and 9 head CTs performed ≤ 48 hours postoperatively.

Temporal changes in pregnancy-associated glycoproteins across different stages of gestation in the Barbari goat.

The objective of this study was to characterize the temporal profile of pregnancy-associated glycoproteins (PAGs; isoforms 1-11) across different stages of gestation in the Barbari goat. Placentae were collected from local abattoir, classified according to crown rump length of the corresponding foetus into five groups (0-30, 31-60, 61-90, 91-120, and 121-150 days of gestation), and used for relative quantification of mRNA expression by Pfaffl method. In addition, adult female goats (pregnant, n = 7; non-pregnant, n = 5) were used to estimate weekly plasma PAG and progesterone (P4) concentrations.

Synthesis and anticancer activity of novel spiro-isoxazoline and spiro-isoxazolidine derivatives of α-santonin.

In the present study, novel spiro derivatives of α-santonin were prepared and tested for their anticancer activity against a panel of six human cancer cell lines. Spiro-isoxazoline and spiro-isoxazolidine derivatives have been generated on C-ring of α-santonin (α-methylene-γ-butyrolactone) by the 1,3-dipolar cycloaddition of α-santonin derivative 6 with nitrile oxides 7 and nitrones 9 respectively. Among all, compound 10b″ had shown IC50 of 0.

Expression of leptin and its receptor in corpus luteum during estrous cycle in buffalo (Bubalus bubalis).

Leptin is supposed to play a crucial role in ovarian luteal dynamics. The present study was aimed to investigate the importance of leptin and its receptors in buffalo corpus luteum (CL) obtained from different stages of the estrous cycle. Real-time RT-PCR (qPCR), western blot and immunohistochemistry techniques were applied to investigate mRNA expression, protein expression and localization of examined factors.

Urological complications in ureteric stenting live related renal transplantation.

Objective: To determine the frequency and types of post-transplant urological complications in live-related kidney transplantation with reference to the impact of JJ ureteric stent.

Study Design: Case series.

Place And Duration Of Study: Shaikh Zayed Postgraduate Medical Institute and National Institute of Kidney Diseases, Lahore, from June 2006 to July 2010.