Susceptibility of sperm chromatin to acid denaturation in situ: A study in endogenous FSH-deprived adult male bonnet monkeys (Macaca radiata)

Acid denaturation of calf thymus DNA in vitro followed by acridine orange (AO) binding induced a 112% increase in the emission of red, a 58% decrease in green, and a consequential decrease in the ratio of green:red fluorescences from 1.7 to 0.9. This metachromatic property of AO on binding to DNA following acid denaturation was utilized to study the susceptibility of normal and ovine follicle-stimulating hormone (oFSH) actively immunized bonnet monkey spermatozoa voided throughout the year. For analyses, the scattergram generated by the emission of red and green fluorescences by 10,000 AO-bound sperm from each semen sample was divided into 4 quadrant zones representing percentage cells fluorescing high green-low red (Q1), high green-high red (Q2), low green-low red (Q3) and low green-high red. (Q4). Normal monkey sperm obtained during the months of July-December exhibited 76, 13, and 11% cells in Q2, Q3, and Q4 quadrants, respectively. However, during January-June, when the females of the species are markedly subfertile, noncycling, and amenorrhoeic, the spermatozoa ejaculated by the male monkeys exhibited 38, 39, and 23% sperm in Q2, Q3, and Q4, respectively, the differences being highly significant (p < .01-.001). FSH deprivation induced significant shifts in fluorescence emissions, from respective controls, with 39, 33, and 28% cells in Q2, Q3, and Q4, respectively, during July-December, and 15, 48, and 37% sperm in Q2, Q3, and Q4 quadrants, respectively, during January-June. It is postulated that the altered kinetics of germ cell transformations and the deficient spermiogenesis observed earlier following FSH deprivation in these monkeys may have induced the enhanced susceptibility to acid denaturation in sperm.

Respuesta al tratamiento super ovulatorio, con Folltropin-V (analogo sintetico de la hormona foliculoestimulante-FSH), en hembras bovinas donantes, de las razas Pardo Suizo y Reyna, en fincas de la Universidad Nacional Agraria, Managua

El presente trabajo investigativo fue realizado en las fincas Las Mercedes y Santa Rosa, ambas propiedad de la Universidad Nacional Agraria, ubicadas en el departamento de Managua-Nicaragua. El objetivo de la presente investigación fué conocer si hay influencia del factor raza en la respuesta súper ovulatoria una vez aplicado el tratamiento hormonal súper ovulatorio FOLLTROPIN-V (análogo sintético de la hormona foliculoestimulante-FSH) en hembras bovinas donantes de las razas Pardo Suizo y Reyna. El tamaño requerido de la muestra fue de 10 hembras donantes para cada raza en estudio; las cuales fueron previamente seleccionadas según la calidad genética de cada hembra, la edad comprendida entre 3-5 años y los resultados del examen clínico y ginecológico que se les realizó para saber si cumplían con los requisitos ó parámetros de salud estimados para esta técnica de mejoramiento productivo. Los datos se obtuvieron mediante el conteo de cuerpos lúteos y folículos a través de palpación rectal de los ovarios; el mismo día en que se realizó la recolecta de los embriones; los cuales fueron llevados al laboratorio para ser clasificados según su estado de desarrollo y calidad, posteriormente fueron transferidos ó congelados. Los resultados obtenidos a ravés del análisis de la respuesta superovulatoria, indican que no existe una diferencia significativa del factor raza, sin embargo se observó que la raza Reyna se comportó mejor que la raza Pardo Su izo al dar mayor cantidad de cuerpos lúteos (R: 14.75 - PS: 11.5) y embriones (R: 9.25 – PS: 5.6), pero sí se atribuye a la raza, la calidad y el estado de desarrollo embrionario que se obtuvieron, siendo la raza Reyna la que obtuvo mejor calidad de embriones aunque por ser la que más embriones dió, también fue la que brindó mayor cantidad de embriones degenerados y oocitos infértiles que son intransferibles.

Repuesta al tratamiento súper ovulatorio, con folltropin V (Análogo sintético de la FSH), en hembras bovinas donantes, de las razas Pardo Suizo y Holsteins de la finca las mercedes de la Universidad Nacional Agraria, Managua

El presente trabajo investigativo se llevo a cabo en la finca Las Mercedes, propiedad de la Universidad Nacional Agraria ubicada en el Departamento de Managua, Nicaragua, en el Km 12.5 Carretera Norte. El objetivo del estudio fue determinar si hay influencia del factor raza en la repuesta súper ovulatoria en las hembras bovinas donantes de la> razas Holsteins y Pardo Suizo sometidas al tratamiento hormonal mediante la aplicación del Folltropin V (Análogo Sintético de la Hormona Foliculoestimulante FSH).El tamallo requerido de la muestra fue d 1 O hembras donantes para cada raza en estudio ;las cuales fueron previamente seleccionados según la calidad genética de cada hembra , la edad comprendida para la selección fue de entre 3 y 5 años además se realizaron exámenes clínicos y ginecológicos para determinar si cumplían con los requisitos o parámetros de salud para realizar esta técnica de mejoramiento productivo, los datos se obtuvieron mediante el conteo de cuerpos lúteos y folículos atravez de la palpación rectal de los ovarios ;el mismo dla en que realizo la recolecta de los embriones; los cuales fueron llevados al laboratorio para ser clasificados según su estado de desarrollo y calidad posteriormente fueron transferidos y congelados . Los resultados obtenidos atravez del análisis de la respuesta súper ovulatoria, indican que hay diferencia signicativa en cuanto al numero de cuerpos híteos en la raza Pardo Suizo ya que obtuvo un promedio de 9.2 cuerpo lúteos y la raza Holsteins obtuvo un promedio de 6.9 cuerpos lúteos . Por otra parte no hubo diferencia significativa en lo que respecta al número de embriones que tuvo un promedio de 3.8 cultivados en comparación con los 3. 7 embriones de la Raza Holsteins.

年龄和FSH在肉牛活体取卵过程中对卵泡和卵母细胞发育的影响

青年牛(12月龄)和经产牛(7-8岁)平均卵泡数(≥2mm)及>10mm卵泡数明显高于老年牛(≥14岁)(P<0.01)。青年牛和经产牛平均卵母细胞数及平均COC数高于老年牛(P<0.05)。1和2级卵母细胞比例青年牛高于或等于经产牛高于老年牛,3和4级卵母细胞比例老年牛高于经产牛高于或等于青年牛(P<0.05)。青年牛和经产牛2级卵母细胞占优势,而老年牛3级卵母细胞居多。不同剂量外源性FSH诱导后,经产牛和老年牛平均卵泡数在各剂量组之间没有显著差异(P>0.05);但≥6mm卵泡数及2级卵母细胞比例明显增加,而2-5mm卵泡数相应减少及裸卵和老化卵比例降低(P<0.05)。在10次取卵过程中,青年牛和经产牛发情当天平均卵母细胞数均较以后各次的数量低(P<0.05)。

Molecular and expression characterization of three gonadotropin subunits common alpha, FSH beta and LH beta in groupers

A SMART cDNA plasmid library was constructed from protogyous greasy grouper (Epinephelus coioides) pituitary, and the full-length cDNAs of three gonadotropin (GTH) subunits common alpha, FSH beta and LH beta were cloned and sequenced from the library. The nucleotide sequences of common alpha, FSH beta and LH beta subunit cDNAs are 647, 594 and 574 bp in length, and encode for mature peptides of 94, 99 and 115 aa, respectively. High homology was observed by amino acid sequence alignment and identity comparison of the grouper mature peptides of common alpha, FSH beta and LH beta with that of other fishes. Phylogenetic tree analyses of the three GTH mature subunits revealed similar phylogeny relationships among the studied fish species. Three polyclonal antibodies were prepared from the in vitro expressed common alpha, FSH beta and LH beta mature proteins, respectively. Western blot analysis and immunofluoresence localization were performed on two typical stages of ovarian development stages in red-spotted grouper. Significant differences in protein expression levels of three gonadotropin subunits were revealed between the two ovarian development stages. In the individuals with resting ovary, common alpha was almost not detected in pituitaries, and FSH beta and LH beta expression levels were very low. While in the individuals with developing ovary, the expression of all three gonadotropin subunits reached to a high level. Immunofluoresence localization indicated that the grouper FSH beta cells mainly distributed in the middle area of PPD, while the LH beta cells distributed more widely, including in the area similar to the FSH beta cells and at the external periphery of pituitary near to the PI side. The common alpha might be expressed in both FSH beta and LH beta cells. Double immunofluoresence localization further demonstrated FSH beta and LH beta expression in distinct cells in the PPD area, although the FSH beta and LH beta cells were detected in the identical area of PPD. (c) 2005 Elsevier Ireland Ltd. All rights reserved.

Studies on equine milk and comparative studies on equine and bovine milk systems

The composition of equine milk differs considerably from that of the milk of the principal dairying species, i.e., the cow, buffalo, goat and sheep. Because equine milk resembles human milk in many respects and is claimed to have special therapeutic properties, it is becoming increasingly popular in Western Europe, where it is produced on large farms in several countries. Equine milk is considered to be highly digestible, rich in essential nutrients and to possess an optimum whey protein:casein ratio, making it very suitable as a substitute for bovine milk in paediatric dietetics. There is some scientific basis for the special nutritional and health-giving properties of equine milk but this study provides a comprehensive analysis of the composition and physico-chemical properties of equine milk which is required to fully exploit its potential in human nutrition. Quantification and distribution of the nitrogenous components and principal salts of equine milk are reported. The effects of the high concentration of ionic calcium, large casein micelles (~ 260 nm), low protein, lack of a sulphydryl group in equine β-lactoglobulin and a very low level of κ-casein on the physico-chemical properties of equine milk are reported. This thesis provides an insight into the stability of equine casein micelles to heat, ethanol, high pressure, rennet or acid. Differences in rennet- and acid-induced coagulation between equine and bovine milk are attributed not only to the low casein content of equine milk but also to differences in the mechanism by which the respective micelles are stabilized. It has been reported that β-casein plays a role in the stabilization of equine casein micelles and proteomic techniques support this view. In this study, equine κ-casein appeared to be resistant to hydrolysis by calf chymosin but equine β-casein was readily hydrolysed. Resolution of equine milk proteins by urea-PAGE showed the multi-phosphorylated isoforms of equine αs- and β-caseins and capillary zone electrophoresis showed 3 to 7 phosphorylated residues in equine β-casein. In vitro digestion of equine β-casein by pepsin and Corolase PP™ did not produce casomorphins BCM-5 or BCM-7, believed to be harmful to human health. Electron microscopy provided very clear, detailed images of equine casein micelles in their native state and when renneted or acidified. Equine milk formed flocs rather then a gel when renneted or acidified which is supported by dynamic oscillatory analysis. The results presented in this thesis will assist in the development of new products from equine milk for human consumption which will retain some of its unique compositional and health-giving properties.

Explorations in a meaning making system: Kerry as an entrepreneurial platform and the equine sector as a cluster

This Portfolio is about the changes that can be supported and achieved through transformational education that impacts on personal, professional and organisational levels. Having lived through an era of tremendous change over the second half of the twentieth century and into the twenty-first the author has a great drawing board to contemplate in the context of professional career experience as an engineer. The ability to engage in ‘subject-object’ separation is the means by which Kegan (1994, 2009) explains that transformation takes place and the Essays in this Portfolio aim to support and bring about such change. Exploration of aspects of ‘Kerry’ is the material selected to both challenge support change in the way of knowing from being subject to certain information and knowledge that to being able to consider it more objectively. The task of being able to distance judgement about the economy and economic development of Kerry was facilitated by various readings around of a number of key thinkers including Kegan, Drucker, Porter and Penrose. The central themes of Kerry or the potential for economic development are built into each Essay. Essay One focuses on reflections of Kerry life - on Kerry people within and without Kerry - and events as they affected understandings of how people related to and worked with one another. These reflections formed the basis for transformational goals identified which required a shift from an engineering mindset to encompass an economics-based view. In Essay Two knowledge of economic concepts is developed by exploring the writings of Drucker, Penrose, and Porter with pertinence to considering economic development generally, and for Kerry in particular in the form of an ‘entrepreneurial platform’. The concepts and theories were the basis of explorations presented in Essays Three and Four. Essay Three focuses on Kerry’s potential for economic development give its current economic profile and includes results from interviews with selected businesses. Essay Four is an exercise in the application of Porter’s ‘Cluster’ concept to the equine sector.

Biochemical characterization of the equine arteritis virus helicase suggests a close functional relationship between arterivirus and coronavirus helicases.

The arterivirus equine arteritis virus nonstructural protein 10 (nsp10) has previously been predicted to contain a Zn finger structure linked to a superfamily 1 (SF1) helicase domain. A recombinant form of nsp10, MBP-nsp10, was produced in Escherichia coli as a fusion protein with the maltose-binding protein. The protein was partially purified by affinity chromatography and shown to have ATPase activity that was strongly stimulated by poly(dT), poly(U), and poly(dA) but not by poly(G). The protein also had both RNA and DNA duplex-unwinding activities that required the presence of 5' single-stranded regions on the partial-duplex substrates, indicating a 5'-to-3' polarity in the unwinding reaction. Results of this study suggest a close functional relationship between the arterivirus nsp10 and the coronavirus helicase, for which NTPase and duplex-unwinding activities were recently demonstrated. In a number of biochemical properties, both arterivirus and coronavirus SF1 helicases differ significantly from the previously characterized RNA virus SF1 and SF2 enzymes. Thus, the combined data strongly support the idea that nidovirus helicases may represent a separate group of RNA virus-encoded helicases with distinct properties.

Expression of cyclooxygenase isoforms in equine gastric ulcers

Mémoire numérisé par la Division de la gestion de documents et des archives de l'Université de Montréal.