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G Elamaran KP Singh MK Singh SK Singla MS Chauhan RS Manik P Palta 《Reproduction in domestic animals》2012,47(6):1027-1036
This study examined the effects of O2 concentration (5% vs 20%) during in vitro maturation (IVM), fertilization (IVF) and culture (IVC) or supplementation of IVM and IVC media with cysteamine (50 and 100 μm , respectively; IVM, IVF and IVC carried out in 20% O2), on blastocyst rate and relative mRNA abundance of some apoptosis‐related genes measured by real‐time qPCR in immature and in vitro‐matured buffalo oocytes and in embryos at 2‐, 4‐, 8‐ to 16‐cell, morula and blastocyst stages. The blastocyst rate was significantly higher (p < 0.05) while the percentage of TUNEL‐positive cells was significantly lower (p < 0.05) under 5% O2 than that under 20% O2. The mRNA expression of anti‐apoptotic genes BCL‐2 and MCL‐1 was significantly higher (p < 0.05) and that of pro‐apoptotic genes BAX and BID was lower (p < 0.05) under 5% O2 than that under 20% O2 concentration at many embryonic stages. Following cysteamine supplementation, the blastocyst rate and the relative mRNA abundance of BCL‐XL and MCL‐1 was significantly higher (p < 0.05) and that of BAX but not BID was lower (p < 0.05) at many stages of embryonic development, although it did not affect the percentage of TUNEL positive cells in the blastocysts significantly. The mRNA expression pattern of these genes during embryonic development was different in 5% vs 20% O2 groups and in cysteamine supplemented vs controls. At the 8‐ to 16‐cell stage, where developmental block occurs in buffalo, the relative mRNA abundance of BCL‐2 and MCL‐1 was highest under 5% O2 concentration and that of BAX and BID was highest (p < 0.05) under 20% O2 concentration. These results suggest that one of the mechanisms through which beneficial effects of low O2 concentration and cysteamine supplementation are mediated during in vitro embryo production is through an increase in the expression of anti‐apoptotic and a decrease in the expression of pro‐apoptotic genes. 相似文献
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Comparative Expression Analysis of Gametogenesis‐Associated Genes in Foetal and Adult Bubaline (Bubalus bubalis) Ovaries and Testes
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SM Shah N Saini S Ashraf M Zandi MK Singh RS Manik SK Singla P Palta MS Chauhan 《Reproduction in domestic animals》2015,50(3):365-377
This study was conducted to identify and analyse the expression of gametogenesis‐associated genes and proteins in foetal and adult buffalo gonads of both the sexes. Relative quantification of the genes was determined by qPCR and Western blotting. Immunohistochemistry was also performed for various gametogenesis‐associated proteins in foetal and adult gonads of both the sexes. We observed significantly (p < 0.05) increased expression of primordial germ cell‐specific, meiotic as well as genes associated with oocyte maturation and development in foetal ovaries as compared to the adult ones. However, significantly (p < 0.05) increased expression of proteins associated with oocyte maturation like GDF9 and ZP4 was found in adult ovaries, indicating temporal regulation of mRNA translation during oogenesis. Meiotic genes showed significantly (p < 0.05) increased expression in adult testes as compared to foetal testes and ovaries, indicating onset of meiosis at a later stage in spermatogenesis. In general, the expression of primordial germ cell‐associated as well as meiotic genes was higher in adult testes, indicating the increased biological activity in the organ. Immunohistochemistry revealed localized expression of gametogenesis‐associated proteins in ovarian follicles and seminiferous tubules of testes, while the surrounding somatic tissues were devoid of these proteins. The study gives an understanding of the sequential and temporal events of gene expression as well as mRNA translation during male and female gametogenesis. It could also be concluded that follicles and seminiferous tubules are the functional units of the female and male gonads, respectively, and their function could be enhanced by appropriate chemical and genetic intervention of the somatic tissue immediately surrounding them. This assumes importance in the context that buffalo attains sexual maturity at an older age of 2–3 years and have smaller ovaries with lesser number of primordial follicles in comparison with cattle, which is suggested to be the main reason of their poor breeding performance. 相似文献
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Noninvasive blood pressure measurement is now a standard diagnostic test in small animal practice. Factors such as patient selection, environment, technical support staff, and necessary equipment and supplies will be discussed. Step by step techniques for obtaining reliable blood pressure measurements in both cats and dogs are presented. 相似文献
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Expression and localization of angiopoietin family in corpus luteum during different stages of oestrous cycle and modulatory role of angiopoietins on steroidogenesis,angiogenesis and survivability of cultured buffalo luteal cells
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SR Mishra MS Parmar VP Yadav R Reshma J Bharati MK Bharti A Paul VS Chouhan G Taru Sharma G Singh M Sarkar 《Reproduction in domestic animals》2016,51(6):855-869
The objective of this study was to document the expression and localization of angiopoietin (ANGPT) family members comprising of angiopoietin (ANGPT1 and ANGPT2), and their receptors (Tie1 and Tie2) in buffalo corpus luteum (CL) obtained from different stages of the oestrous cycle, and the modulatory role of ANGPT1 and ANGPT2 alone or in combinations on progesterone (P4) secretion and mRNA expression of phosphotidylinositide‐3kinase‐protein kinase B (PI3K‐AKT), phosphoinositide‐dependent kinase (PDK), protein kinase B (AKT), Bcl2 associated death promoter (BAD), caspase 3 and von willebrand factor (vWF) in luteal cells obtained from midluteal phase (MLP) of oestrous cycle in buffalo. Real‐time RT‐PCR (qPCR), Western blot and immunohistochemistry were applied to investigate mRNA expression, protein expression and localization of examined factors whereas, the P4 secretion was assessed by RIA. The mRNA and protein expression of ANGPT1 and Tie2 was maximum (p < .05) in mid luteal phase (MLP) of oestrous cycle. The ANGPT2 mRNA and protein expression was maximum (p < .05) in early luteal phase, decreased in MLP and again increased in late luteal phase of oestrous cycle. ANGPT family members were localized in luteal cells and endothelial cells with a stage specific immunoreactivity. P4 secretion was highest (p < .05) with 100 ng/ml at 72 hr when luteal cells were treated with either protein alone. The mRNA expression of PDK, AKT and vWF was highest (p < .05) and BAD along with caspase 3 were lowest (p < .05) at 100 ng/ml at 72 hr of incubation period, when cultured luteal cells were treated with either protein alone or in combination. To conclude, our study explores the steroidogenic potential of angiopoietins to promote P4 secretion, luteal cell survival and angiogenesis through an autocrine and paracrine actions in buffalo CL. 相似文献
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This retrospective case study aims to evaluate the accuracy of menace response, response to nasal stimulation and proprioceptive placing in diagnosing forebrain lesion in dogs. A total of 145 client-owned dogs investigated by magnetic resonance imaging study of the brain between December 2017 and June 2019 were evaluated. Seventy-one dogs with no magnetic resonance imaging-detectable intracranial and significant cerebrospinal fluid abnormality or recent history of seizure (<48 h) served as controls. Binary regression analysis was performed to determine the sensitivity, specificity and likelihood ratios of each selected test. Older age at presentation was a significant risk factor for the presence of a forebrain lesion. Menace (62.5%) and proprioceptive deficits (40.5%) were common findings in all dogs. They were also significantly associated with the presence of forebrain abnormality. Moreover, they were more sensitive (77.3% and 82.2%, respectively) and specific (50.0% and 62.5%, respectively) when applied to dogs aged 6 years or older. Nonetheless, all of these tests' likelihood ratios, and thus reliability are poor. These neurological tests are commonly employed for diagnosing forebrain disease in dogs, yet are not highly accurate in diagnosing forebrain abnormality. Clinicians should interpret these clinical test results along with the patient history when designing a diagnostic plan. 相似文献
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beta-Adrenergic blockers, particularly atenolol, are often recommended for the tachycardia and hypertension that accompany hyperthyroidism; however, the effects of monotherapy with atenolol on both systolic blood pressure (SBP) and heart rate (HR) have not been reported. Twenty hyperthyroid cats with SBP > or = 160 mmHg were studied retrospectively to investigate the SBP and HR lowering effects of atenolol. Median pre-treatment SBP and HR for all cats were 186.5 mmHg and 231 beats/min, respectively. All cats were treated with atenolol at a dosage of 1-2 mg/kg PO q 12h for a minimum of 5 days prior to reassessment and treatment with radioactive iodine. SBP and HR both decreased following atenolol therapy in this group of cats to median values of 171.5 mmHg (P=0.0088) and 185/min (P=0.0003). However, when successful clinical control of hypertension was defined as a post-treatment SBP < 160 mmHg, atenolol monotherapy was ineffective in 70% of the cases. There was no statistically significant difference in baseline serum total thyroxine or atenolol dosage between clinical responders and non-responders. While atenolol effectively reduces HR in most cats with hyperthyroidism, elevated SBP is poorly controlled, and the addition of another vasodilator such as amlodipine or an angiotensin converting enzyme inhibitor is needed to treat associated hypertension. 相似文献
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Prothrombin time and activated partial thromboplastin time using a point‐of‐care analyser (Abaxis VSpro®) in Bennett's wallabies (Macropus rufogriseus)
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BN Nevitt SK Chinnadurai MK Watson JN Langan MJ Adkesson 《Australian veterinary journal》2016,94(10):384-386
There are few reports of coagulation times in marsupial species. Blood samples collected from 14 Bennett's wallabies (Macropus rufogriseus) under anaesthesia during routine health assessments were analysed for prothrombin time (PT) and activated partial thromboplastin time (aPTT) using a point‐of‐care analyser (POC) (Abaxis VSPro®). The wallabies had an aPTT mean of 78.09 s and median of 78.1 s. The PT for all wallabies was greater than 35 s, exceeding the longest time measured on the POC. Although PT was significantly longer, aPTT was similar to the manufacturer's domestic canine reference range. 相似文献