Timed artificial insemination in beef cattle using GnRH agonist, PGF2alpha and estradiol benzoate (EB)

The present work evaluated low-cost protocols for timed artificial insemination (TAI) in beef cattle. In Experiment 1, cycling nonlactating Nelore cows (Bos indicus, n=98) were assigned to the following groups: GnRH-PGF (GP) and GnRH-PGF-GnRH (GPG), whereas cycling (n=328, Experiment 2) or anestrus (n = 225, Experiment 3) lactating (L) cows were divided into 3 groups: GP-L, GPG-L and GnRH-PCF-Estradiol benzoate (GPE-L). In Experiment 4, lactating cows (n=201) were separated into 3 groups: GP-L, GPE-L and G 1/2PE-L. Animals from Experiment 1, 3 and 4 were treated (Day 0), at random stages of the estrous cycle, with 8 mug of buserelin acetate (GnRH agonist) intramuscularly (im), whereas in Experiment 2 half of the cows received 8 and the other half 12 mug of GnRH (im). Seven days later (D 7) all animals were treated with 25 mg of dinoprost trometamine (PGF2 alpha, im) except those cows from the G 1/2PE-L group which received only 1/2 dose of PGF2 alpha (12.5 mg) via intravulvo-submucosa (ivsm). Alter PGF2 alpha injection the animals from the control groups (GP and GP-L) were observed twice daily to detect estrus and Al was performed 12 h afterwards. The cows from the other groups received a second GnRH injection (D 8 in GPG-L and d9 in GPG groups) or one injection of estradiol benzoate (EB, 1.0 mg, D 8 in GPE-L group). All cows from GPG and GPG-L or GPE-L groups were AI 20 to 24 or 30 to 34 h, respectively, after the last hormonal injection. Pregnancy was determined by ultrasonography or rectal palpation 30 to 50 days after AI. In the control groups (GP and GP-L) percentage of animals detected in heat (44.5 to 70.3%) and pregnancy rate (20 to 42%) varied according to the number of animals with corpus luteum (CL) at the beginning of treatment. The administration of a second dose of GnRH either 24 (Experiment 2) or 48 h (Experiment 1) after PGF2 alpha resulted in 47.7 and 44.9% pregnancy rates, respectively, after TAI in cycling animals. However, in anestrus cows the GPG treatment induced a much lower pregnancy rate (14.9%) after TAI. The replacement of the second dose of GnRH by EB (GPE-L) resulted in a pregnancy rate (43.3%) comparable to that obtained after GnRH treatment (GPG-L, 47.7%, Experiment 2). Furthermore, the use of 1/2 dose of PGF2 alpha (12.5 mg ivms, Experiment 4) resulted in pregnancy rate (43.5%) similar to that observed with the full dose (im). Both protocols GPG and GPE were effective in synchronizing ovulation in cycling Nelore cows and allowed approximately a 45% pregnancy rate after TAI. Additionally, the GPE treatment is a promising alternative to the use of GPG in timed Al of beef cattle, due to the low cost of EB when compared to GnRH agonists. (C) 2001 by Elsevier B.V.

Synchronization of ovulation in crossbred dairy heifers using gonadotrophin-releasing hormone agonist, prostaglandin F2a and human chorionic gonadotrophin or estradiol benzoate

Girolando (Gir x Holstein) is a very common dairy breed in Brazil because it combines the rusticity of Gir (Bos indicus) with the high milk yield of Holstein (Bos taurus). The ovarian follicular dynamics and hormonal treatments for synchronization of ovulation and timed artificial insemination were studied in Girolando heifers. The injection of a gonadotrophin-releasing hormone (GnRH) agonist was followed 6 or 7 days (d) later by prostaglandin F2a (PGF2a). Twenty-four hours after PGF2a injection either human chorionic gonadotropin (hCG, GPh-d6 and GPh-d7 groups) or estradiol benzoate (EB, GPE-d6 and GPE-d7 groups) was administered to synchronize ovulation and consequently allow timed artificial insemination (AI) 24 and 30 h after hCG and EB injection, respectively. Follicular dynamics in Girolando heifers was characterized by the predominance of three follicular waves (71.4%) with sizes of dominant follicles (10-13 mm) and corpus luteum (approximately 20 mm) similar to those for Bos indicus cattle. In the GnRH-PGF-hCG protocol, hCG administration induced earlier ovulation (67.4 h, P<0.01) compared to the control group (GnRH-PGF) and a better synchronization of ovulation, since most of it occurred within a period of 12 to 17 h. Pregnancy rate after timed AI was 42.8 (3/7, GPh-d6) to 50% (7/14, GPh-d7). In contrast, estradiol benzoate (GnRH-PGF-EB protocol) synchronized ovulation of only 5 of 11 heifers from the GPE-d7 group and of none (0/7) from the GPE-d6 group, which led to low pregnancy rates after timed AI (27.3 and 0%, respectively). However, since a small number of Girolando heifers was used to determine pregnancy rates in the present study, pregnancy rates should be confirmed with a larger number of animals.

Differential response of the luteal phase and fertility in cattle following ovulation of the first-wave follicle with human chorionic gonadotropin or an agonist of gonadotropin-releasing hormone

A series of experiments with Holstein heifers was conducted to develop the capability of inducing accessory corpus luteum (CL) with a GnRH agonist (Buserelin, 8 mu g; GnRHa) or hCG; (3,000 IU) to increase plasma progesterone concentrations (Exp. 1, 2, and 3) and to test whether induction of accessory CL with hCG will increase conception rates in heifers (Exp. 4) and lactating cows (Exp. 5). In Exp. 1, heifers were treated on d 5 after estrus with GnRHa (n = 8) or saline (n = 7); heifers in Exp. 2 received hCG (n = 5) or saline (n = 4) on d 5. Experiment 3 allowed a contemporary evaluation of heifers treated on d 5 with GnRHa (n = 6), hCG (n = 6), saline (n = 6), or GnRHa at d 5 and hCG at the time of the induced ovulation (n = 5). The GnRHa and hCG were equally effective in inducing an accessory CL (93% induction rate), but the subsequent increase in progesterone concentrations was greater in hCG-treated heifers. A greater half life of hCG may provide longer LH-like stimulation of the first-wave follicle and subsequent developing accessory CL or a greater luteotropic effect on the original CL. Induction of an accessory CL with hCG on d 5 or 6 after insemination did not increase pregnancy rates in fertile heifers (Exp. 4: hCG = 64.8% vs control = 62.9%; n = 243) or lactating dairy cows during summer heat stress (Exp. 5: hCG = 24.2% vs control = 23.5%; n = 201).

Similar anxiolytic-like effects following intra-amygdala infusions of benzodiazepine receptor agonist and antagonist: Evidence for the release of an endogenous benzodiazepine inverse agonist in mice exposed to elevated plus-maze test

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Embryo recovery and pregnancy rates after the delay of ovulation and fixed time insemination in superstimulated beef cows

The aim of this study was to evaluate the effect of delaying ovulation subsequent to superstimulation of follicular growth in beef cows (Bos indicus) on embryo recovery rates and the capacity of embryos to establish pregnancies. Ovulation was delayed by three treatments using either progesterone (CIDR-B®) or a GnRH agonist (deslorelin). Multiparous Nelore cows (n = 24) received three of four superstimulation treatments in an incomplete block design (n = 18 per group). Cows in Groups CTRL, P48 and P60 were treated with a CIDR-B device plus estradiol benzoate (EB, 4 mg, i.m.) on Day-5, while cows in Group D60 were implanted with deslorelin on Day-7. Cows were superstimulated with FSH (Folltropin-V® 200 mg), from Day 0 to 3, using twice daily injections in decreasing amounts. All cows were treated with a luteolytic dose of prostaglandin on Day 2 (08:00 h). CIDR-B devices were removed as follows: Group CTRL, Day 2 (20:00 h); Group P48, Day 4 (08:00 h); Group P60, Day 4 (20:00 h). Cows in Group CTRL were inseminated at 10, 20 and 30 h after first detected estrus. Ovulation was induced for cows in Group P48 (Day 4, 08:00 h) and Groups P60 and D60 (Day 4, 20:00 h) by injection of LH (Lutropin®, 25 mg, i.m.), and these cows were inseminated 10 and 20 h after treatment with LH. Embryos were recovered on Days 11 or 12, graded and transferred to synchronized recipients. Pregnancies were determined by ultrasonography around Day 100. Data were analyzed by mixed procedure, Kruskal-Wallis and Chi-square tests. The number of ova/embryos, transferable embryos (mean ± S.E.M.) and pregnancy rates (%) were as follows, respectively: Group CTRL (10.8 ± 1.8, 6.1 ± 1.3, 51.5), P48 (12.6 ± 1.9, 7.1 ± 1.0, 52.3), P60 (10.5 ± 1.6, 5.7 ± 1.3, 40.0) and D60 (10.3 ± 1.7, 5.0 ± 1.2, 50.0). There were no significant differences among the groups (P > 0.05). It was concluded that fixed time AI in association with induced ovulation did not influence embryo recovery. Furthermore, pregnancy rates in embryos recovered from cows with delayed ovulation were similar to those in embryos obtained from cows treated with a conventional superstimulation protocol. © 2002 Elsevier B.V. All rights reserved.

Proteinase-activated receptor-2 (PAR2) agonist causes periodontitis in rats

Proteinase-activated receptor-2 (PAR2) is a G-protein-coupled receptor that mediates cellular responses to extracellular proteinases. Since PAR2 is expressed by oral epithelial cells, osteoblasts, and gingival fibroblasts, where its activation releases interleukin-8, we hypothesized that PAR2 activation may participate in periodontal disease in vivo. We investigated the role of PAR2 activation in periodontal disease in rats. Radiographic and enzymatic (myeloperoxidase) analysis revealed that topical application of PAR2 agonist causes periodontitis but also exacerbates existing periodontitis, leading to significant alveolar bone loss and gingival granulocyte infiltration. Inhibition of matrix metalloproteinase (MMP) and cyclo-oxygenase (COX) decreased PAR2 agonist-induced periodontitis. More specifically, the overexpression of COX-1, COX-2, MMP-2, and MMP-9 in gingival tissues suggests that they are involved in PAR 2-induced periodontitis. In conclusion, PAR2 agonist causes periodontitis in rats through a mechanism involving prostaglandin release and MMP activation. Inhibition of PAR2 may represent a novel approach to modulate host response in periodontitis.

Analysis of agonist and antagonist effects on thyroid hormone receptor conformation by hydrogen/deuterium exchange

Thyroid hormone receptors (TRs) are ligand-gated transcription factors with critical roles in development and metabolism. Although x-ray structures of TR ligand-binding domains (LBDs) with agonists are available, comparable structures without ligand (apo-TR) or with antagonists are not. It remains important to understand apo-LBD conformation and the way that it rearranges with ligands to develop better TR pharmaceuticals. In this study, we conducted hydrogen/deuterium exchange on TR LBDs with or without agonist (T 3) or antagonist (NH3). Both ligands reduce deuterium incorporation into LBD amide hydrogens, implying tighter overall folding of the domain. As predicted, mass spectroscopic analysis of individual proteolytic peptides after hydrogen/ deuterium exchange reveals that ligand increases the degree of solvent protection of regions close to the buried ligand-binding pocket. However, there is also extensive ligand protection of other regions, including the dimer surface at H10-H11, providing evidence for allosteric communication between the ligand-binding pocket and distant interaction surfaces. Surprisingly, Cterminal activation helix H12, which is known to alter position with ligand, remains relatively protected from solvent in all conditions suggesting that it is packed against the LBD irrespective of the presence or type of ligand. T 3, but not NH3, increases accessibility of the upper part of H3-H5 to solvent, and we propose that TR H12 interacts with this region in apo-TR and that this interaction is blocked by T 3 but not NH3.Wepresent data from site-directed mutagenesis experiments and molecular dynamics simulations that lend support to this structural model of apo-TR and its ligand-dependent conformational changes. (Molecular Endocrinology 25: 15-31, 2011). Copyright © 2011 by The Endocrine Society.

Cost-effectiveness comparison between pituitary down-regulation with a gonadotropin-releasing hormone agonist short regimen on alternate days and an antagonist protocol for assisted fertilization treatments

Objective: To compare cost-effectiveness between pituitary down-regulation with a GnRH agonist (GnRHa) short regimen on alternate days and GnRH antagonist (GnRHant) multidose protocol on in vitro fertilization (IVF)/intracytoplasmic sperm injection (ICSI) outcome. Design: Prospective, randomized. Setting: A private center. Patient(s): Patients were randomized into GnRHa (n = 48) and GnRHant (n = 48) groups. Intervention(s): GnRHa stimulation protocol: administration of triptorelin on alternate days starting on the first day of the cycle, recombinant FSH (rFSH), and recombinant hCG (rhCG) microdose. GnRHant protocol: administration of a daily dose of rFSH, cetrorelix, and rhCG microdose. Main Outcome Measure(s): ICSI outcomes and treatment costs. Result(s): A significantly lower number of patients underwent embryo transfer in the GnRHa group. Clinical pregnancy rate was significantly lower and miscarriage rate was significantly higher in the GnRHa group. It was observed a significant lower cost per cycle in the GnRHa group compared with the GnRHant group ($5,327.80 ± 387.30 vs. $5,900.40 ± 472.50). However, mean cost per pregnancy in the GnRHa was higher than in the GnRHant group ($19,671.80 ± 1,430.00 vs. $11,328.70 ± 907.20). Conclusion(s): Although the short controlled ovarian stimulation protocol with GnRHa on alternate days, rFSH, and rhCG microdose may lower the cost of an individual IVF cycle, it requires more cycles to achieve pregnancy. Clinical Trial Registration Number: NCT01468441. © 2013 by American Society for Reproductive Medicine.

Avaliação da taxa de prenhez em novilhas tratadas com protocolo GnRH-PGF2'alfa' -Benzoato de Estradiol (GPE) e em vacas submetidas a tratamentos hormonais com progesterona associados ou não a remoção temporária de bezerros

Pós-graduação em Medicina Veterinária - FMVZ

Induction of multiple ovulations in mares using low doses of GnRH agonist Deslorelin Acetate at 48 hours after luteolysis

This study was aimed to test low doses of a GnRH agonist, deslorelin acetate (DA), for induction of multiple ovulations in mares and to determine its impact upon their reproductive efficiency. Seven mares aging from 8-20 years were used in three consecutive reproductive cycles. Mares were initially monitored by ultrasound irrespectively of cycle stage, inseminated and submitted to embryo collection (EC) (T1). Immediately after, mares received 7.5 mg dinoprost tromothamine (DT) and were monitored by ultrasound twice a day until larger follicle reached 23-25mm and the second >18mm (T2). At this time point, mares received 100 mu g DA and ovulation was induced with 1000 mu g DA and 1000IU hCG when largest follicle reached 33-35mm in diameter, followed by EC. Mares were further allocated to T3 when received 7.5 mg DT after EC on 12 and 100 mu g DA 48 h later. DA treatment was performed until dominant follicle reached 34 +/- 1 mm or 6 days of application. All EC were performed 8 days after ovulation. Mares with multiple ovulations in T1, T2 and T3 were 14.28% (1/7), 100.00% (7/7) and 0.00% (0/7), respectively, and averaged 0.43 +/- 0.53 in T1, 0.86 +/- 0.38 in T2 and 0.00 in T3 embryos per donor, respectively. Embryo recovery rate was 43.00% in T1, 85.71% in T2 and 0.00% T3. In conclusion, use of DA in mares with follicles larger than 25mm enhanced dominant and co-dominant follicle growth, that ultimately increased the incidence of multiple ovulations and embryo recovery rate.

Effects of 15d-PGJ(2)-loaded poly(D,L-lactide-co-glycolide) nanocapsules on inflammation

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

CONTROL of THE CENTRAL CHEMOREFLEX BY A5 NORADRENERGIC NEURONS IN RATS

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

Ractopamine in diets for finishing gilts

Foi realizado um experimento utilizando-se 468 fêmeas, com peso inicial de 84,77 ± 7,20 kg, alojadas em 36 baias, para avaliar os efeitos da adição de ractopamina nas dietas sobre o desempenho, as características e os rendimentos de cortes comerciais da carcaça, a composição e os cortes cárneos do pernil. O desenho experimental usado foi o de blocos ao acaso com quatro níveis de ractopamina (0, 5, 10 e 15 mg/kg de dieta) e nove repetições com 13 animais por unidade experimental. O critério para formação dos blocos foi o peso inicial dos animais. Não foram observados efeitos dos níveis de ractopamina sobre o ganho de peso diário dos animais. O consumo diário de ração apresentou redução linear com o aumento do nível de ractopamina nas dietas. A conversão alimentar dos animais melhorou de forma linear com a inclusão de ractopamina na dieta dos animais. Verificaram-se também diferenças significativas na proporção de gordura e carne na carcaça e nos cortes cárneos. Houve efeito linear decrescente da ractopamina sobre a quantidade e profundidade de gordura e efeito linear crescente sobre a porcentagem de carne magra, de carne no pernil e sobre os pesos de semimembranosus e gluteus medius. Dietas para fêmeas suínas em fase final de terminação devem conter 15 mg de ractopamina por kg.

Serotonergic neurons in the median raphe nucleus regulate inhibitory avoidance but not escape behavior in the rat elevated T-maze test of anxiety

Rationale: A wealth of evidence supports the involvement of the serotonergic neurons of the median raphe nucleus (MRN) in anxiety. However, it is presently unclear whether serotonergic pathways arising from this nucleus play distinguishing regulatory roles in defensive behaviors that have been associated with specific subtypes of anxiety disorders. Objectives: To evaluate the role of the MRN serotonergic neurons in the regulation of two defensive behaviors, inhibitory avoidance and escape, which have been related, respectively, to generalized anxiety and panic disorders. Methods: Male Wistar rats were submitted to the elevated T-maze test of anxiety after intra-MRN administration of drugs that either non-selectively or selectively change the activity of the serotonergic neurons. Results: Intra-MRN injection of FG 7142 (0.04 and 0.08 nmol) and kainic acid (0.03 and 0.06 nmol), drugs that non-selectively stimulate the MRN serotonergic neurons, facilitated inhibitory avoidance acquisition, but impaired escape performance. Microinjection of muscimol (0.11 and 0.22 nmol), a compound that non-selectively inhibits the activity of the MRN serotonergic neurons, impaired inhibitory avoidance and facilitated escape performance. Both kainic acid and muscimol also changed rat locomotion in the open-field test. Intra-MRN injection of 8-OH-DPAT (0.6-15 nmol) and WAY-100635 (0.18-0.74 nmol), respectively an agonist and an antagonist of somatodendritic 5-HT1A receptors located on serotonergic neurons of the MRN, only affected inhibitory avoidance-while the former inhibited the acquisition of this behavior, the latter facilitated it. Conclusion: MRN serotonergic neurons seem to be selectively involved in the regulation of inhibitory avoidance in the elevated T-maze. This result supports the proposal that 5-HT pathways departing from this nucleus play an important role in anxiety processing, with implications for pathologies such as generalized anxiety disorder.