RELATIONSHIP BETWEEN PLASMA 17-BETA ESTRADIOL ON THE DAY OF ESTRUS AND NUMBER OF VIABLE EMBRYOS IN BOS-INDICUS (NELLORE) COWS SUPEROVULATED WITH FSH

Thirteen cows, Bos indicus, of the Nellore breed were superovulated with 22 mg of follicle stimulating hormone (FSH) administered by intramuscular route during four consecutive days (D10, D11, D12 and D13), starting on the 10th day of the estrous cycle (day 0 = estrus). Prostaglandin (PGF2alpha, 1.0 mg, im) was administered on D12, 48 h after the first FSH injection, for the induction of estrus on D14, when artificial insemination was performed. Seven days later (D21 of the cycle), embryos were collected, and evaluated, and the ovarian response was estimated on the basis of number of corpora lutea determined by rectal palpation. Blood samples were obtained for the determination of plasma 17-beta estradiol on D10, D11, D12, D13, D14 and D21 and plasma progesterone on D14 by RIA. The donors were divided into two groups according to progesterone levels on D14, the day of the induced estrus (GI: P4 less-than-or-equal-to 1.00 ng/ml, N = 5 and GII: P4 > 1.00 ng/ml, N = 8). A linear positive correlation was observed between plasma 17-beta estradiol concentration on the day of estrus and viable embryo number. We conclude that plasma 17-beta estradiol and progesterone concentrations on the day of estrus can be used to predict the viability of embryos recovered from Nellore cows superovulated with FSH.

Fertility-associated proteins in Nelore bull sperm membranes

The present study was undertaken to evaluate the protein composition of the sperm membranes (SM) of Nelore bulls, assessing protein markers associated with bull fertility, and whether these markers can be used for predicting bull fertility. Samples were obtained of 20 Nelore bulls, with fertility ranked and divided into three groups (greater, normal and least). To rank the bull's fertility weighted classification was used (according to the number of pregnant cows, number of AI cows and number of herds, considering three different breeding seasons), using the PROC GENMOD as a statistical model, with 99% significance. A total of 7897 Nelore cows, randomly distributed among 28 different farms, were considered in the statistical analyses. The bulls were divided into three fertility groups (pregnancy rates): greater (%F > 80), normal (79 <%F > 71) and least (< 68%F) with 3, 13 and 4 bulls, respectively. Two-dimensional gel electrophoresis (2DE) of sperm membranes indicated in 27 spots (SM40, SM53, SM69, SM93, SM102, SM111, SM137, SM138, SM189, SM196, SM201, SM202, SM204, SM225, SM236, SM237, SM239, SM241, SM246, SM247, SM275, SM283, SM342, SM346, SM355, SM372, SM391) was prevalent in the higher fertility group, and just one spot (SM244) was prevalent in the lower fertility group. Spots SM244 and SM239 had their identification defined by PMF/MALDI-MS, as BSP-A3 and aSFP, respectively. Both these proteins showed a great potential for predicting bull's fertility. The amount of aSFP was 8.5 times greater in the sperm membrane protein profile of the higher fertility groups of Nelore bulls. Besides that, the BSP-A3 was 2.5 times greater in the lower fertility group. For the other spots potentially associated with fertility not yet identified, additional tests will be necessary, but it is clear that the 2D electrophoresis of the sperm membrane can be used for a new approach to predict Nelore bull fertility. (c) 2005 Elsevier B.V. All rights reserved.

Proteins of Leishmania (Viannia) shawi confer protection associated with Th1 immune response and memory generation

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Ultrastructural analysis of bovine oocytes from ovarian follicles with different diameters

In vitro embryo production is an important technique for facilitating the reproduction of animals with high genetic merit. The greatest challenge for the reproducibility of this technique is the quality of the oocyte that is submitted for in vitro maturation. The aim of this work was to evaluate the ultrastructural characteristics of oocytes from follicles of different diameters using transmission electron microscopy. The animals were divided into 2 groups and were given a single i.m. injection of 250 IU FSH (Pluset, Serono, Italy). To synchronize the follicular growth, all follicles > 2 mm were aspirated during the estrous cycle, which was considered day zero (D0). Group 1 (G1; n = 4) received FSH on day 1 (D1), and the 2- to 5-mm follicles were aspirated on day 2 (D2). The animals in group 2 (G2; n = 5) received FSH on day 2 (D2), and their 10- to 15-mm follicles were aspirated on day 5 (D5). After aspiration, the oocytes from both groups were fixed and prepared for ultrastructural analysis. The oocytes analyzed from both groups had similar ultrastructural characteristics. The presence and distribution of organelles in the cytoplasm of the oocytes did not differ between groups, suggesting that, in relation to the ultrastructural characteristics, oocytes from 2 to 5 mm and 10 to 15 mm follicles are similar.

Morfologia do figado da paca (Cuniculus Paca, Linnaeus 1766)

The elements related to the morphology of the liver of paca (Cuniculus paca), the second largest rodent of the Brazilian fauna, were observed; this species present zootechnical potential. Eight animals from the animals sector of Faculdade de Ciencias Agrarias e Veterinarias - Campus of Jaboticabal - UNESP, which is duly certified by IBAMA as an experimental breeding institute, were used. Through a dissection procedure, it was found that the liver of the paca is located in the cranial portion of the abdomen, immediately after the diaphragm, to which it is connected by the triangular, coronary, and falciform ligaments, having its bigger part located right to the medium plan. The liver of this rodent presents the following lobation: right lateral lobe, right medial lobe, quadrate lobe, left medial lobe, and left lateral lobe, besides the caudate lobe formed by the papillary process of caudate lobe and the caudate process of caudate lobe. Gallbladder is located between the quadrate and right medial lobes. Fragments of this organ were collected, fixed, and histologically prepared, being the samples analyzed through light microscopy. It was microscopically observed that intralobular connective tissue is scarce, basically it consists polyhedral hepatocytes organized into cords interposed between sinusoids and the portal triads are found in the lobe, consisting of the portal vein, hepatic artery, and biliary duct.

Comprimento da peça intermediária de espermatozóides bovinos e suas relações com características do sêmen e fertilidade

With the objective of identifying predictors of fertility in bulls, the correlation coefficients of the midpiece length of bovine spermatozoa with semen traits and fertility were estimated. Data were obtained from semen samples of 50 crossbred bulls submitted to a progeny test. The midpiece length was determined in 40 midpieces of each bull through a image analysis system (Videoplan). Besides the physical traits before and after semen freezing, the oxygen consumption rate and the cytochemical activity index were observed. The correlation coefficients were low and non-significant (P> 0.05), indicating that the midpiece length is not a good predictor of fertility.

Evaluation of plasma membrane integrity of frozen-thawed goat spermatozoa with or without seminal plasma

The aim of the present work was to evaluate plasma membrane integrity, motility, vigor and morphology of fresh and frozen goat spermatozoa with or without seminal plasma. Semen samples were diluted in Tris solution, before and after thawing, with a combination of carboxifluorescein diacetate and propidium iodide. The results showed differences (P < 0.01) for motility and minor defects in the presence or absence of seminal plasma, for both fresh and frozen samples. Periods of collection had a significant effect on motility, probably due to changes in the photoperiod. Plasma membrane integrity was significantly reduced by the freezing process, whether seminal plasma was present or absent. In conclusion, removal of seminal plasma decreased motility and vigor rates in frozen samples. The photoperiod probably decreased the testosterone level, contributing negatively to the high percentage of sperm abnormalities, mainly damaged membranes. The use of fluorescent probes allowed a better estimation of the percentage of functional cells, instead of only estimating the percentage of motile cells or morphology defects. © 2001 Elsevier Science B.V. All rights reserved.

Desenvolvimento morfolôgico dos ovários em embriões e fetos bovinos da raça Nelore

The morphologic events related to the prenatal development of the ovaries in 81 Nelore breed embryos and fetuses gathered in a local slaughterhouse, with age range from 26 to 240 days following fecundation were studied. The age of fetuses was estimated from measures taken in the cranium-caudal direction. The sex was identified from macroscopic observations and using Polymerase Chain Reaction (PCR) technique. For histology the gonads were fixed in Bourn's fluid for 24 hours and 5 μm thick section's were stained with hematoxylin-eosin. Formation of gonadal ridge and presence of germinal cells were found within it at 29 and 34 days, respectively. Oogonia and primordial follicles, unlike the growing follicles, exhibited significant differences in diameter in the various periods studied. Positive correlation (P<0.05) was found between the diameter of oogonia and their nucleus as well as between primordial and growing follicles with their oocytes and respective nuclei. The gonad was fully formed at 40 days. Primordial follicles, in the growing stage, and antral follicles first appeared, approximately at 95, 140, and 180 days, respectively. Despite the onset and duration of oogenesis being similar to that of taurine breeds, folliculogenesis initiates at an early stages in the Nelore breed.

Efeitos de diferentes sistemas de cultivo in vitro sobre o crescimento de folículos pré-antrais isolados de ovários de fetos bovinos

The objective of this study is to use different in vitro culture systems of preantral follicles from Nelore breed bovine fetuses in the last gestation quarter. The evaluation of treatments considered the time of growth of isolated follicles. Preantral follicles were mechanically isolated and submitted to the individual culture, for 9 days, in media no supplemented or supplemented with fetal calf serum (FCS), bovine serum albumin (BSA) or synthetic defined supplement substitute of serum KnockoutSR (KNO). We have also evaluated the effects of collagen gel or fetal calf fibroblast monolayer as substratum for in vitro cultures. The increase on the follicular diameter was followed in the first day (0 h), at the 72 h, 144 h and 216 h. Considering cultures of isolated follicles, the results have shown that the association between media supplemented with FCS and collagen gel was significantly more efficient on the increase of the follicular diameter than other treatments. It is not still established a system of appropriate cultivation that sustains the differentiation and multiplication of the granular cells and that maintains the contact of the same ones with the oocyte to provide molecules and factors that supply the metabolic demand. We also understand that our results also represent another promising step on the search for the ultimate system of in vitro culture of preantral follicles from bovines.

Morfogênese do testículo de embriões e fetos de vacas da raça nelore (Bos taurus indicus)

The aim of this study was to accompany the process of testicular development from the non-differentiable phase to its complete formation. Embryos and fetuses of Nelore breed cows (Bos Taurus indicus) were obtained in slaughterhouses near the Uberlandia city, Minas Gerais. The gonads and the embryos were fixed in Bouin's fixative and afterwards processed for conventional optical microscopy. The gonadal was observed firstly in a 1.0 cm long embryo. In 2.5 cm long embryos the presence of the albuginea allows the sex identification. The mean thickness of the albuginea ranged from 29.08 to 558.45 mm. Gradually increase of vascularization of the albuginea and parenchyma is observed. The mediastinum is located centrally. There was a decrease in the space occupied by the testicular cords, from 63.71 to 41.99% of the total testes volume. Its diameter ranged from 31.68 to 48.80 mm. The diameter of germinal cells (and their nuclei) was from 12.27 (6.65) to 16.95 914.21) mm. The quantity of germinal cells by cross section of cord decreased from a maximum of 2.80 to 0.76. The total number of germinal cells was from 16 at the beginning of colonization of the gonad to 18.32 x 106 at the end of the study. The number of Sertoli's cells by cross section of cord ranged from 10.00 to 16.25. The results obtained show that the origin and formation of testes in embryos and fetuses from Nelore breed cows (Bos taurus indicus) does occur in a very similar way to what is described for Bos taurus taurus.

γ-Radiation induces apoptosis via sarcoplasmatic reticulum in guinea pig ileum smooth muscle cells

We investigated the effects of γ-radiation on cells isolated from the longitudinal smooth muscle layer of the guinea pig ileum, a relatively radioresistant tissue. Single doses (up to 50 Gy) reduced the amount of sarcoplasmatic reticulum and condensed the myofibrils, as shown by electron microscopy 3 days post-irradiation. After that, contractility of smooth muscle strips was reduced. Ca2+ handling was altered after irradiation, as shown in fura-2 loaded cells, with elevated basal intracellular Ca2+, reduced amount of intrareticular Ca2+, and reduced capacitive Ca2+ entry. Radiation also induced apoptosis, judged from flow cytometry of cells loaded with proprium iodide. Electron microscopy showed that radiation caused condensation of chromatin in dense masses around the nuclear envelope, the presence of apoptotic bodies, fragmentation of the nucleus, detachment of cells from their neighbors, and reductions in cell volume. Radiation also caused activation of caspase 12. Apoptosis was reduced by the administration of the caspase inhibitor Z-Val-Ala-Asp-fluoromethyl-ketone methyl ester (Z-VAD-FMK) during the 3 day period after irradiation, and by the chelator of intracellular Ca2+, 1,2-bis(o-aminophenoxy)-ethane-N,N,N′,N′-tetraacetic acid (BAPTA), from 1 h before until 2 h after irradiation. BAPTA also reduced the effects of radiation on contractility, basal intracellular Ca2+, amount of intrareticular Ca2+, capacitative Ca2+ entry, and apoptosis. In conclusion, the effects of gamma radiation on contractility, Ca2+ handling, and apoptosis appear due to a toxic action of intracellular Ca2+. Ca2+-induced damage to the sarcoplasmatic reticulum seems a key event in impaired Ca2+ handling and apoptosis induced by γ-radiation. © 2008 Elsevier B.V. All rights reserved.

Los 20 años del MERCOSUR: una integración a dos velocidades

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Acute exercise decreases PTP-1B protein level and improves insulin signaling in the liver of old rats

It is now commonly accepted that chronic inflammation associated with obesity during aging induces insulin resistance in the liver. In the present study, we investigated whether the improvement in insulin sensitivity and insulin signaling, mediated by acute exercise, could be associated with modulation of protein-tyrosine phosphatase 1B (PTP-1B) in the liver of old rats. Aging rats were subjected to swimming for two 1.5-h long bouts, separated by a 45 min rest period. Sixteen hours after the exercise, the rats were sacrificed and proteins from the insulin signaling pathway were analyzed by immunoblotting. Our results show that the fat mass was increased in old rats. The reduction in glucose disappearance rate (Kitt) observed in aged rats was restored 16 h after exercise. Aging increased the content of PTP-1B and attenuated insulin signaling in the liver of rats, a phenomenon that was reversed by exercise. Aging rats also increased the IRβ/PTP-1B and IRS-1/PTP-1B association in the liver when compared with young rats. Conversely, in the liver of exercised old rats, IRβ/PTP-1B and IRS-1/PTP-1B association was markedly decreased. Moreover, in the hepatic tissue of old rats, the insulin signalling was decreased and PEPCK and G6Pase levels were increased when compared with young rats. Interestingly, 16 h after acute exercise, the PEPCK and G6Pase protein level were decreased in the old exercised group. These results provide new insights into the mechanisms by which exercise restores insulin signalling in liver during aging. © 2013 Moura et al; licensee BioMed Central Ltd.