RESUMO

Reviewing the current state of knowledge on reproductive performance and productive traits in rams has many advantages. First, the compilation of this information will serve as a literature resource for scientists conducting research around the world and will contribute to the understanding of the data collected and interpreted by researchers on the different hormonal strategies used to improve reproductive performance in rams. Second, it will allow scientists to identify current knowledge gaps and set future research priorities in ram reproduction. Rams play an important role in the global flock economy, but their reproductive analysis has been limited in the use of hormonal technologies to increase the productivity of sheep flocks. In this review, we cite the most important works on six hormones that, in one way or another, modify the hypothalamus-pituitary-gonadal axis, at different doses, in and out of the reproductive season, breeds, application methods, among other factors. The overall aim is to increase the reproductive efficiency of rams in different scenarios and, in some cases, of other species due to the lack of limited information on rams.

RESUMO

In this study, we aimed to propose changes in the protocol of cultured Astyanax altiparanae hypophysation to increase the maximum ovulation rate of 60% registered previously. To that two consecutive experiments were conducted. In the first experiment, three carp pituitary homogenate (CPH) doses (3, 6, and 9 mg/kg) were administered in a single injection, while in the second experiment, the 6 mg/kg CPH dose was tested either in single or double injections. In the first experiment, a single injection of 3 mg/kg CPH did not induce final oocyte maturation or spawning, while a dose of 6 mg/kg CPH resulted in an increase in the plasma level of prostaglandin (PGF2α) at ovulation. The single higher dose of 9 mg/kg CPH did not improve reproductive performance and even though anticipated the resumption of meiosis it was detrimental to the spawning rate. In the second experiment, the dose of 6 mg/kg CPH fractionated into two injections led to a higher spawning rate, spawning volume per female body mass, frequency of post-ovulatory complexes, and PGF2α concentration at ovulation compared to the single injection. The most effective treatment remained the 6 mg/kg of CPH fractionated into two injections, but still providing very low proportion of ovulated females (∼40 %). Overall, this study indicates that the spawning protocols for this species need to be improved to induce ovulation in a larger number of females and be more potent in those females that respond positively.

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Background: Prostaglandin F2 alpha (PGF2 α) binds to the specific receptor (PTGFR) on the corpus luteum (CL) in mammals, inducing regression of the CL structure (luteolysis) and initiating a new cycle. While PGF2 α is effective only on mature CL, the immature CL structure (early luteal phase) resists PGF2 α. In this study, sildenafil citrate, which is used to increase blood flow in the genital organs for treating specific pregnancy issues in women, was administered during the early luteal phase in a rabbit model to test the hypothesis of enhancing blood flow to the CL, thereby promoting earlier maturation and enabling a response to PGF2 α. Materials, Methods & Results: The study was conducted in 2 sub-studies: clinical and molecular. A large number of rabbits were initially included in the sub-studies to ensure a sufficient number of pseudo-pregnant rabbits. Ovulation in rabbits was induced with buserelin acetate and was considered as day 0 of the study. The sub-studies were continued with rabbits whose pseudo-pregnancies were confirmed according to progesterone (P4 ) results. As a result, the studies were continued with a total of 41 pseudo-pregnant New Zealand female rabbits, 21 of which were included in the clinical sub-study and 20 in the molecular sub-study. In both sub-studies, on day 3 of the luteal period, rabbits in the treatment group received 5 mg/kg sildenafil citrate and all rabbits received a single dose of exogenous PGF2 α on day 4 to induce luteolysis. In the clinical sub-study, echotexture and intraovarian blood flow changes in the ovaries were determined by ultrasonography (USG) examination. In the molecular sub-study, the expression changes of Hypoxia Inducible Factor 1 Alpha (HIF1A) and Vascular Endothelial Growth Factor (VEGF) related to angiogenesis, Steroidogenic Acute Regulatory Protein (StAR) related to P4 metabolism, Prostaglandin-Endoperoxide Synthase 2 (PTGS2) related to prostaglandin (PG) mechanism and 15-Hydroxyprostaglandin Dehydrogenase (HPGD) genes at mRNA level were determined using Real Time Polymerase Chain Reaction (RT-PCR) in CL tissues obtained with ovariohysterectomy (OVH) at 1 and 12 h after PGF2 α injection. In addition, blood samples were collected for determine P4 levels from all rabbits. In the clinical sub-study; there was no difference between the groups in mean gray values (MGV), whereas there was a significant decrease in both pulsatile index (PI) and resistance index (RI) values at 40 min after PGF2 α injection (P < 0.05). In the molecular sub-study, it was determined that sildenafil citrate had no significant effect (P > 0.05) on the expression levels 1 and 12 h after PGF2 α injection. According to the results of the molecular sub-study, no significant effect of sildenafil citrate on the mRNA expression levels in the investigated genes was detected (P > 0.05). However, within each group, differences were found according to OVH time after PGF2 α injection. It was observed that PTGS2 and HPGD mRNA expressions decreased at the 12th h compared to the 1st h, while HIF1A expression increased (P < 0.05). Discussion: According to the results obtained from clinical and molecular sub-studies, it was determined that a single dose of sildenafil citrate (5 mg/kg) applied on the 3rd day of the luteal period did not contribute to the maturation process of the CL, did not increase blood flow, and was insufficient to break the resistance of the CL against PGF2 α applied on the 4th day of the luteal period. However, a significant decrease in the PI value at the 40th min after PGF2 α injection suggests that sildenafil citrate has a supportive effect, and that this decrease is also seen in the RI value, suggesting that its effect is insufficient against the vasoconstrictive effect of PGF2 α.

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Although studies have shown positive effects of gonadotropin releasing hormone (GnRH) or prostaglandin F2α (PGF2α) at the moment of fixed-time artificial insemination (FTAI) in the conception rate (CR) of cattle, its effects on treatments based on progesterone (P4) and estradiol benzoate (EB) is still not conclusive. The objective of this study was (1) to evaluate the effect of a PGF2α analogue at FTAI in the CR of crossbred beef cows submitted to a 11d FTAI protocol based on P4 and EB; and (2) to describe the CR between PGF2α-treated and control cows in different body condition scores (BCS) and parity categories. Crossbred (½ Nellore and ½ Angus) beef cows were submitted to a synchronization protocol and randomly assigned into 2 groups: Control (n = 163), at FTAI cows received 2 mL of saline solution as a placebo, and PGF2α (n = 163), at FTAI cows were treated with PGF2α analogue (10 mg of dinoprost tromethamine). Pregnancy diagnosis was performed 33d post-FTAI. Binary logistic regression model was used to analyze the effect of PGF2α treatment on CR. There was no difference in CR between PGF2α and control groups (P > 0.05; odds ratio (OR) = 0.92; confidence interval (CI) = 0.59-1.4). A greater CR was found in heifers (P = 0.0006, OR = 2.65, CI = 1.61 - 4.38) and multiparous (P = 0.0006, OR = 2.12, CI = 1.04 - 4.3) when compared to primiparous cows. Cows with low BCS (4; 9-point scale) showed lower CR when compared with moderate BCS (5-6; 9-point scale) (P < 0.05; OR = 0.10; CI = 0.06 - 0.18). There was no numerical difference on CR between PGF2α-treated and control cows in different BCS and parity categories. The results suggested that the CR in this study was not influenced by 10 mg PGF2α analogue at FTAI.

RESUMO

Although studies have shown positive effects of gonadotropin releasing hormone (GnRH) or prostaglandin F2α (PGF2α) at the moment of fixed-time artificial insemination (FTAI) in the conception rate (CR) of cattle, its effects on treatments based on progesterone (P4) and estradiol benzoate (EB) is still not conclusive. The objective of this study was (1) to evaluate the effect of a PGF2α analogue at FTAI in the CR of crossbred beef cows submitted to a 11d FTAI protocol based on P4 and EB; and (2) to describe the CR between PGF2α-treated and control cows in different body condition scores (BCS) and parity categories. Crossbred (½ Nellore and ½ Angus) beef cows were submitted to a synchronization protocol and randomly assigned into 2 groups: Control (n = 163), at FTAI cows received 2 mL of saline solution as a placebo, and PGF2α (n = 163), at FTAI cows were treated with PGF2α analogue (10 mg of dinoprost tromethamine). Pregnancy diagnosis was performed 33d post-FTAI. Binary logistic regression model was used to analyze the effect of PGF2α treatment on CR. There was no difference in CR between PGF2α and control groups (P > 0.05; odds ratio (OR) = 0.92; confidence interval (CI) = 0.59-1.4). A greater CR was found in heifers (P = 0.0006, OR = 2.65, CI = 1.61 - 4.38) and multiparous (P = 0.0006, OR = 2.12, CI = 1.04 - 4.3) when compared to primiparous cows. Cows with low BCS (4; 9-point scale) showed lower CR when compared with moderate BCS (5-6; 9-point scale) (P < 0.05; OR = 0.10; CI = 0.06 - 0.18). There was no numerical difference on CR between PGF2α-treated and control cows in different BCS and parity categories. The results suggested that the CR in this study was not influenced by 10 mg PGF2α analogue at FTAI.(AU)

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The release of endometrial prostaglandin-F2α (PGF2α) in bovine females can be induced in vivo by estradiol (E2). However, its role in this mechanism has not been clarified. We hypothesized that E2 stimulates the activity and abundance of protein kinase C (PKC) and phospholipase A2 (PLA2). Our objective in this study was to analyze the effects of PKC and PLA2 inhibitors on PGF2α synthesis induced by E2 and calcium ionophore (CI) in bovine endometrial cells (BEND cells; Experiment 1). Additionally, we evaluated the abundance of PKC and PLA2 in endometrial explants of cows treated or not with E2 17 days after estrus (D17, D0 = estrus; Experiment 2). In Experiment 1, BEND cells were submitted to a PKC inhibitor (10 µM of C25H24N4O2; bisindolylmaleimide I, or BIS I), a PLA2 inhibitor (20 µM of arachydoniltrifluoromethane or AACOCF3), or none. The BEND cells were subsequently treated with E2 and CI, and PGF2α concentrations were measured in the culture medium through radioimmunoassay. For DIF-12 (PGF2α concentration 12 h after treatment subtracted from PGF2α concentration at hour 0), no PKC inhibitor effect was observed (P= 0.2709). However, DIF-12 was lower (P < 0.05) for groups treated with the PLA2 inhibitor and PLA2 inhibitor + CI + E2 groups than the control and CI + E2 groups. Thus, AACOCF3 was an efficient PLA2 inhibitor in the BEND cells culture system, and E2 did not stimulate the synthesis of PKC and PLA2. In Experiment 2, cyclic Nellore heifers received none (n = 5) or 3 mg (n = 6) of 17ß-E2 on D17 and were slaughtered 2 h after administration. The abundance of PKC and PLA2 in the endometrial tissue was evaluated using Western blotting analysis. No E2 effect was observed on PKC (P = 0.08) and PLA2 (P = 0.56). We concluded that E2 did not stimulate the activity and abundance of PKC and PLA2.(AU)

A liberação endometrial de prostaglandina-F2α (PGF2α) em fêmeas bovinas pode ser induzida in vivo pelo estradiol (E2). Entretanto o seu mecanismo de ação ainda não foi bem esclarecido. Nossa hipótese é que o E2 estimula a atividade e a abundância da proteína quinase C (PKC) e da fosfolipase A2 (PLA2). Nosso objetivo com este estudo foi analizar os efeitos de inibidores de PKC e PLA2 na síntese de PGF2α induzida por E2 e ionóforo de cálcio (CI) em células endometriais bovinas (células BEND; Experimento 1). Adicionalmente, nós avaliamos a abundância de PKC e PLA2 em explantes endometriais de vacas tratadas com ou sem E2 17 dias após o estro (D17, D0 = estro; Experimento 2). No Experimento 1, células BEND foram submetidas ao inibidor de PKC (10 µM de C25H24N4O2; bisindolylmaleimide I, ou BIS I), e ao inibidor de PLA2 (20 µM de arachydoniltrifluoromethane ou AACOCF3) ou a nenhum inibidor. As células BEND foram subsequentemente tratadas com E2 e CI e concentrações de PGF2α foram mensuradas no meio de cultura por radioimunoenssaio. Para DIF-12 (concentração de PGF2α 12 horas depois do tratamento, subtraída da concentração de PGF2α na hora 0), não foi observado efeito do inibidor de PKC (P = 0.2709). Entretanto DIF-12 foi menor (P < 0.05) nos grupos tratados com inibidor de PLA2 e inibidor de PLA2 + CI + E2 quando comparados com o grupo controle e o grupo CI + E2. O AACOCF3 foi um eficiente inibidor de PLA2 em sistema de cultura de células BEND e o E2 não estimulou a síntese de PKC e PLA2. No Experimento 2, novilhas Nelore cíclicas receberam 3 mg de 17ß-E2 (n = 6) ou nenhum tratamento (n = 5) no D17 e foram abatidas duas horas depois da administração dos tratamentos. A quantidade de PKC and PLA2 no tecido endometrial foi avaliada pela técnica de Western Blotting. Não foi observado efeito do E2 sobre a PKC (P= 0.08) e nem sobre a PLA2 (P= 0.56). Conclui-se que o E2 não estimulou a atividade e abundância de PKC e PLA2.(AU)

Assuntos

RESUMO

The release of endometrial prostaglandin-F2α (PGF2α) in bovine females can be induced in vivo by estradiol (E2). However, its role in this mechanism has not been clarified. We hypothesized that E2 stimulates the activity and abundance of protein kinase C (PKC) and phospholipase A2 (PLA2). Our objective in this study was to analyze the effects of PKC and PLA2 inhibitors on PGF2α synthesis induced by E2 and calcium ionophore (CI) in bovine endometrial cells (BEND cells; Experiment 1). Additionally, we evaluated the abundance of PKC and PLA2 in endometrial explants of cows treated or not with E2 17 days after estrus (D17, D0 = estrus; Experiment 2). In Experiment 1, BEND cells were submitted to a PKC inhibitor (10 µM of C25H24N4O2; bisindolylmaleimide I, or BIS I), a PLA2 inhibitor (20 µM of arachydoniltrifluoromethane or AACOCF3), or none. The BEND cells were subsequently treated with E2 and CI, and PGF2α concentrations were measured in the culture medium through radioimmunoassay. For DIF-12 (PGF2α concentration 12 h after treatment subtracted from PGF2α concentration at hour 0), no PKC inhibitor effect was observed (P= 0.2709). However, DIF-12 was lower (P < 0.05) for groups treated with the PLA2 inhibitor and PLA2 inhibitor + CI + E2 groups than the control and CI + E2 groups. Thus, AACOCF3 was an efficient PLA2 inhibitor in the BEND cells culture system, and E2 did not stimulate the synthesis of PKC and PLA2. In Experiment 2, cyclic Nellore heifers received none (n = 5) or 3 mg (n = 6) of 17ß-E2 on D17 and were slaughtered 2 h after administration. The abundance of PKC and PLA2 in the endometrial tissue was evaluated using Western blotting analysis. No E2 effect was observed on PKC (P = 0.08) and PLA2 (P = 0.56). We concluded that E2 did not stimulate the activity and abundance of PKC and PLA2.(AU)

A liberação endometrial de prostaglandina-F2α (PGF2α) em fêmeas bovinas pode ser induzida in vivo pelo estradiol (E2). Entretanto o seu mecanismo de ação ainda não foi bem esclarecido. Nossa hipótese é que o E2 estimula a atividade e a abundância da proteína quinase C (PKC) e da fosfolipase A2 (PLA2). Nosso objetivo com este estudo foi analizar os efeitos de inibidores de PKC e PLA2 na síntese de PGF2α induzida por E2 e ionóforo de cálcio (CI) em células endometriais bovinas (células BEND; Experimento 1). Adicionalmente, nós avaliamos a abundância de PKC e PLA2 em explantes endometriais de vacas tratadas com ou sem E2 17 dias após o estro (D17, D0 = estro; Experimento 2). No Experimento 1, células BEND foram submetidas ao inibidor de PKC (10 µM de C25H24N4O2; bisindolylmaleimide I, ou BIS I), e ao inibidor de PLA2 (20 µM de arachydoniltrifluoromethane ou AACOCF3) ou a nenhum inibidor. As células BEND foram subsequentemente tratadas com E2 e CI e concentrações de PGF2α foram mensuradas no meio de cultura por radioimunoenssaio. Para DIF-12 (concentração de PGF2α 12 horas depois do tratamento, subtraída da concentração de PGF2α na hora 0), não foi observado efeito do inibidor de PKC (P = 0.2709). Entretanto DIF-12 foi menor (P < 0.05) nos grupos tratados com inibidor de PLA2 e inibidor de PLA2 + CI + E2 quando comparados com o grupo controle e o grupo CI + E2. O AACOCF3 foi um eficiente inibidor de PLA2 em sistema de cultura de células BEND e o E2 não estimulou a síntese de PKC e PLA2. No Experimento 2, novilhas Nelore cíclicas receberam 3 mg de 17ß-E2 (n = 6) ou nenhum tratamento (n = 5) no D17 e foram abatidas duas horas depois da administração dos tratamentos. A quantidade de PKC and PLA2 no tecido endometrial foi avaliada pela técnica de Western Blotting. Não foi observado efeito do E2 sobre a PKC (P= 0.08) e nem sobre a PLA2 (P= 0.56). Conclui-se que o E2 não estimulou a atividade e abundância de PKC e PLA2.(AU)

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RESUMO

This study examined the effect of administering an additional dose of prostaglandin F2 alpha (PGF2α), in a fixed-time artificial insemination protocol (FTAI), on the fertility of female Nellore cattle. Two experiments were carried out: the first (Experiment I) took place in the state of Tocantins and the second (Experiment II) in the state of Pará, Brazil. In Experiment I (E1), 80 cows were used in three treatments (T1, T2 and T3) in which all received the same FTAI protocol. In T1 (n = 29), the cows received 12.5 mg of Dinoprost on day 9; in T2 (n = 28), they received the additional dose on day 10; and in T3 (n = 23; control group), the animals did not receive the additional PGF2α dose. Experiment II consisted of 147 bovine females distributed into two treatment groups, namely, T1 - 72 animals receiving the same protocol as T1 of E1; and T2 - 75 animals receiving the same protocol as T3 of E1. Statistical analysis was performed using SAS software, applying the PROC NPARWAY procedure for E1, and means were compared by the Wilcoxon test at the 5% significance level. In Experiment II, the data were subjected to analysis of variance by PROC GLIMMIX and means were compared by the T test at the 5% significance level. The following pregnancy rates were obtained in Experiment I: T1 - 62.06% (18/29); T2 - 57.14% (16/28); and T3 - 52.17% (12/23), with no significant difference observed between treatments. In Experiment II, pregnancy rate in T1 was 66.67% (48/72), whereas in T2 it was 41.33% (31/75), with a significant difference detected (P < 0.05). An additional dose of PGF2α provides an increase in pregnancy rate in Nellore females.(AU)

Objetivou-se avaliar o efeito da administração de uma dose adicional de prostaglandina F2 alfa (PGF2α) em protocolo de inseminação artificial em tempo fixo (IATF) sobre a fertilidade de fêmeas bovinas da raça Nelore. Foram realizados dois experimentos, sendo um (E1) no estado do Tocantins e o segundo (E2) no estado do Pará. No experimento 1 (E1) foram utilizadas 80 fêmeas bovinas distribuídas em três tratamentos (T1, T2 e T3), todos animais receberam o mesmo protocolo de IATF, diferindo no T1 (n=29), que recebeu 12,5 mg de Dinoprost no dia 9, T2 (n=28) recebeu a dose adicional no dia 10 e o T3 (n=23) - Grupo controle, que não recebeu a dose adicional de PGF2α. O experimento 2 (E2) constituído por 147 fêmeas bovinas distribuídas em dois tratamentos. T1: com 72 animais, recebendo mesmo protocolo do T1 do E1. T2: constituído por 75 animais, com protocolo idêntico ao T3 do E1. A análise estatistica foi realizada no programa SAS, sendo no E1 utilizando-se o PROC NPARWAY e as médias comparadas pelo teste de Wilcoxon com nível de significância de 5%. No E2 os dados foram submetidos à análise de variância pelo PROC GLIMMIX e as médias comparadas pelo teste T com nível de significância de 5%. No experimento 1 a taxa de prenhez no T1 foi de 62,06% (18/29), T2 de 57,14% (16/28) e T3 de 52,17% (12/23), não sendo observada diferença significativa entre os tratamentos. No experimento 2 a taxa de prenhez do T1 foi de 66,67% (48/72), enquanto T2 foi de 41,33% (31/75), verificando-se diferença significativa (P < 0,05). A dose adicional de PGF2α promoveu incremento na taxa de prenhez de fêmeas Nelore.(AU)

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BACKGROUND: Prostaglandins (PG) are lipid mediators derived from arachidonic acid metabolism. They are involved in cellular processes such as inflammation and tissue homeostasis. PG production is not restricted to multicellular organisms. Trypanosomatids also synthesize several metabolites of arachidonic acid. Nevertheless, their biological role in these early-branching parasites and their role in host-parasite interaction are not well elucidated. Prostaglandin F2α synthase (PGF2S) has been observed in the Leishmania braziliensis secreted proteome and in L. donovani extracellular vesicles. Furthermore, we previously reported a positive correlation between L. braziliensis PGF2S (LbrPGF2S) expression and pathogenicity in mice. METHODS: LbrPGF2S gene expression and PGF2α synthesis in promastigotes were detected and quantified by western blotting and EIA assay kit, respectively. To investigate LbrPGF2S localization in amastigotes during bone marrow-derived macrophage infection, parasites expressing mCherry-LbrPGF2S were generated and followed by time-lapse imaging for 48 h post-infection. PGF2S homolog sequences from Leishmania and humans were analyzed in silico using ClustalW on Geneious v6 and EMBOSS Needle. RESULTS: Leishmania braziliensis promastigotes synthesize prostaglandin F2α in the presence of arachidonic acid, with peak production in the stationary growth phase under heat stress. LbrPGF2S is a cytoplasmic protein enriched in the secretory site of the parasite cell body, the flagellar pocket. It is an enzyme constitutively expressed throughout promastigote development, but overexpression of LbrPGF2S leads to an increase of infectivity in vitro. The data suggest that LbrPGF2S may be released from intracellular amastigotes into the cytoplasm of bone marrow-derived macrophages over a 48-hour infection period, using time-lapse microscopy and mCherry-PGF2S (mChPGF2S)-expressing parasites. CONCLUSIONS: LbrPGF2S, a parasite-derived protein, is targeted to the host cell cytoplasm. The putative transfer of this enzyme, involved in pro-inflammatory lipid mediator synthesis, to the host cell suggests a potential role in host-parasite interaction and may partially explain the increased pathogenicity associated with overexpression of LbrPGF2S in L. braziliensis. Our data provide valuable insights to help understand the importance of parasite-derived lipid mediators in pathogenesis.

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RESUMO

This study examined the effect of administering an additional dose of prostaglandin F2 alpha (PGF2α), in a fixed-time artificial insemination protocol (FTAI), on the fertility of female Nellore cattle. Two experiments were carried out: the first (Experiment I) took place in the state of Tocantins and the second (Experiment II) in the state of Pará, Brazil. In Experiment I (E1), 80 cows were used in three treatments (T1, T2 and T3) in which all received the same FTAI protocol. In T1 (n = 29), the cows received 12.5 mg of Dinoprost on day 9; in T2 (n = 28), they received the additional dose on day 10; and in T3 (n = 23; control group), the animals did not receive the additional PGF2α dose. Experiment II consisted of 147 bovine females distributed into two treatment groups, namely, T1 - 72 animals receiving the same protocol as T1 of E1; and T2 - 75 animals receiving the same protocol as T3 of E1. Statistical analysis was performed using SAS software, applying the PROC NPARWAY procedure for E1, and means were compared by the Wilcoxon test at the 5% significance level. In Experiment II, the data were subjected to analysis of variance by PROC GLIMMIX and means were compared by the T test at the 5% significance level. The following pregnancy rates were obtained in Experiment I: T1 - 62.06% (18/29); T2 - 57.14% (16/28); and T3 - 52.17% (12/23), with no significant difference observed between treatments. In Experiment II, pregnancy rate in T1 was 66.67% (48/72), whereas in T2 it was 41.33% (31/75), with a significant difference detected (P < 0.05). An additional dose of PGF2α provides an increase in pregnancy rate in Nellore females.

Objetivou-se avaliar o efeito da administração de uma dose adicional de prostaglandina F2 alfa (PGF2α) em protocolo de inseminação artificial em tempo fixo (IATF) sobre a fertilidade de fêmeas bovinas da raça Nelore. Foram realizados dois experimentos, sendo um (E1) no estado do Tocantins e o segundo (E2) no estado do Pará. No experimento 1 (E1) foram utilizadas 80 fêmeas bovinas distribuídas em três tratamentos (T1, T2 e T3), todos animais receberam o mesmo protocolo de IATF, diferindo no T1 (n=29), que recebeu 12,5 mg de Dinoprost no dia 9, T2 (n=28) recebeu a dose adicional no dia 10 e o T3 (n=23) - Grupo controle, que não recebeu a dose adicional de PGF2α. O experimento 2 (E2) constituído por 147 fêmeas bovinas distribuídas em dois tratamentos. T1: com 72 animais, recebendo mesmo protocolo do T1 do E1. T2: constituído por 75 animais, com protocolo idêntico ao T3 do E1. A análise estatistica foi realizada no programa SAS, sendo no E1 utilizando-se o PROC NPARWAY e as médias comparadas pelo teste de Wilcoxon com nível de significância de 5%. No E2 os dados foram submetidos à análise de variância pelo PROC GLIMMIX e as médias comparadas pelo teste T com nível de significância de 5%. No experimento 1 a taxa de prenhez no T1 foi de 62,06% (18/29), T2 de 57,14% (16/28) e T3 de 52,17% (12/23), não sendo observada diferença significativa entre os tratamentos. No experimento 2 a taxa de prenhez do T1 foi de 66,67% (48/72), enquanto T2 foi de 41,33% (31/75), verificando-se diferença significativa (P < 0,05). A dose adicional de PGF2α promoveu incremento na taxa de prenhez de fêmeas Nelore.

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This study was conducted to evaluate effects of two administrations of d-cloprostenol at different intervals to synchronize the time of estrus and ovulation among estrous cyclic goats. In Experiment 1, 32 does were treated with 30 µg d-cloprostenol at 7.5 (T7.5, n = 16) or 11.5-day (T11.5, n = 16) intervals. In Experiment 2, the same treatments were administered and there was AI of the does (T7.5, n = 40 and T11.5, n = 38). In Experiment 1, ultrasonic assessments of ovaries were conducted at the time of the second administration of d-cloprostenol, every 12 h until detection of ovulation, and 7 days after estrous onset to detect the corpora lutea, as well as for pregnancy diagnosis 40 days after AI. In Experiment 1, the estrous response (90.6%, 29/32) was similar (P > 0.05) in both groups. Diameter of the largest follicle at the time of administration of the second dose was larger (P = 0.01) in the T7.5 than T11.5 group (7.0 compared with 5.7 mm), while the values for ovarian variables were similar (P > 0.05). In Experiment 2, the greatest (P < 0.001) synchrony in timing of initiation of estrus in does (T7.5 = 83.3% and T11.5 = 50.0%) occurred after the second day (36-48 h). The pregnancy rate tended (P = 0.0836) to be greater for does in the T7.5 (71.4%, 40/56) than T11.5 (55.6%, 30/54) group. With use of both protocols, there were acceptable estrous synchronization and pregnancy rates in estrous cyclic dairy goats.

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This paper reports an online SPE-LC-MS/MS method for the simultaneous quantification of prostaglandins (PGE2 and PGF2α) in menstrual fluid samples. To meet this goal human peripheral serum was used as surrogate matrix. The analytes were trapped on an OASIS HLB cartridge for 3 min, for sample cleanup and enrichment, and then transferred during only 42 s to an HSS T3 C18 analytical column, for separation and analysis. Prostaglandins (PGs) were detected by selected reaction monitoring in negative ion mode, PGE2 (m/z 351 → 315) and PGF2α (m/z 353 → 193) using isotope-labeled internal standard (PGE2-d4, m/z 355 → 319). The concentration linear range was of 10.34-1.034 ng mL-1 and the lower limit of quantification (LLOQ) was 10.34 ng mL-1 for both PGs. Validation parameters were successfully assessed according to the European Medicines Agency guideline (EMA), also comprising the FDA normative. The method showed no matrix effect and process efficiency around 100%, in addition to only 15 min of analysis time with lower solvent consumption. The method application was carried out using two menstrual fluid sample groups: control (n = 15) and treatment group (n = 7; samples from women that used Tahiti lemon juice). The PGF2α levels were found to be higher in treated group than in control group (p ≤ 0.05), denoting an effect of the intake of Tahiti lemon juice on the menstrual inflammatory process. The on-line method herein reported could be useful for the analysis of PGs from large research studies.

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Prostaglandin F2α (PGF2α) determines luteolysis in cattle, and the ability to manipulate its endogenous synthesis is indispensible for large-scale animal breeding. Estradiol (E2) and progesterone (P4) modulate several molecular pathways in endometrial cells, including the synthesis of PGF2α; however, its specific mechanisms are still not totally known. This study investigated the production in vitro and possible modulation of endometrial PGF2α due to a local effect of endogenous E2 in the ipsilateral uterine horn (UH) containing the dominant follicle (DF) or from P4 in ipsilateral horn containing the corpus luteum (CL). The PGF2α stimulators oxytocin (OT) and phorbol 12,13-dibutyrate (PDBu) were incubated with endometrial explants, and PGF2α content was measured. For that, cycling cows were synchronized, the development of DF and CL was examined by ultrasonography and on the seventh day of the estrous cycle, endometrial explants were collected and cultured in medium supplemented with 10-6 M PDBu or 10-6 M OT or non-supplemented. Media samples were collected immediately after treatment and 60 min later. Radioimmunoassay showed that the PGF2α content of the UH ipsilateral to the DF was 49% less than that of the contralateral UH (8.22 ± 0.95 vs. 12.24 ± 0.95 pg/mL/mg tissue, respectively; P < 0.01). However, the PGF2α levels did not differ between the UHs as a function of the CL position (9.46 ± 0.95 vs. 11 ± 0.95 pg/mL/mg; P > 0.05). The cellular stimulators promoted an increase in PGF2α synthesis (P < 0.02), and the effects differed among the animals (P < 0.04). The PGF2a production was higher in the explants treated with PDBu rather than OT (13.68 ± 1.16 vs. 10.01 ± 1.16 pg/mL/mg tissue, respectively; P < 0.05). In conclusion, PGF2α synthesis is modulated by the presence of the DF (local E2) but not the CL (local P4), and both PDBu and OT stimulated PGF2a synthesis.(AU)

A prostaglandina F2α (PGF2α) determina a luteólise em bovinos. A capacidade de manipular sua síntese endógena é indispensável para a produção animal em grande escala. O estradiol (E2) e a progesterona (P4) modulam diversas vias moleculares das células endometriais, incluindo a síntese de PGF2α; no entanto, pouco se sabe sobre seus mecanismos específicos. Este trabalho investigou a produção in vitro e a possível modulação da PGF2α endometrial devido a um efeito local do E2 endógeno no corno uterino ipsilateral ao folículo dominante (FD) ou da P4 no corno ipsilateral ao corpo lúteo (CL). Os estimuladores de PGF2α oxitocina (OT) e 12,23-dibutirato de forbol (PDBu) foram incubados com explantes endometriais, e o conteúdo de PGF2α foi mensurado. Para tal, vacas cíclicas foram sincronizadas, o desenvolvimento de FD e CL foi examinado por ultrassonografia, e no 17º dia do ciclo estral os explantes endometriais foram coletados e cultivados em meio ou suplementados com PDBu 10-6M ou 10-6M OT. As amostras de meio foram coletadas imediatamente após o tratamento e sessenta minutos depois. O radioimunoensaio mostrou que o conteúdo de PGF2α do corno ipsilateral ao FD foi 49% menor que o do corno contralateral (8,22 ± 0,95 vs. 12,24 ± 0,95 pg/mL/mg de tecido, respectivamente, P < 0,01). No entanto, os níveis de PGF2α não diferiram entre os cornos em função da posição do CL (9,46 ± 0,95 versus 11 ± 0,95 pg/mL/mg; P > 0,05). Os estimuladores celulares promoveram um aumento na síntese de PGF2α (P < 0,02), e os efeitos diferiram entre os animais (P < 0,04). A produção de PGF2α foi maior nos explantes tratados com PDBu em comparação à OT (13,68 ± 1,16 versus 10,01 ± 1,16 pg/mL/mg de tecido, respectivamente, P < 0,05). A conclusão obtida foi que a síntese de PGF2α é: modulada pela presença do FD (E2 local), mas não do CL (P4 local); e estimulada por PDBu e OT.(AU)

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Prostaglandin F2α (PGF2α) determines luteolysis in cattle, and the ability to manipulate its endogenous synthesis is indispensible for large-scale animal breeding. Estradiol (E2) and progesterone (P4) modulate several molecular pathways in endometrial cells, including the synthesis of PGF2α; however, its specific mechanisms are still not totally known. This study investigated the production in vitro and possible modulation of endometrial PGF2α due to a local effect of endogenous E2 in the ipsilateral uterine horn (UH) containing the dominant follicle (DF) or from P4 in ipsilateral horn containing the corpus luteum (CL). The PGF2α stimulators oxytocin (OT) and phorbol 12,13-dibutyrate (PDBu) were incubated with endometrial explants, and PGF2α content was measured. For that, cycling cows were synchronized, the development of DF and CL was examined by ultrasonography and on the seventh day of the estrous cycle, endometrial explants were collected and cultured in medium supplemented with 10-6 M PDBu or 10-6 M OT or non-supplemented. Media samples were collected immediately after treatment and 60 min later. Radioimmunoassay showed that the PGF2α content of the UH ipsilateral to the DF was 49% less than that of the contralateral UH (8.22 ± 0.95 vs. 12.24 ± 0.95 pg/mL/mg tissue, respectively; P < 0.01). However, the PGF2α levels did not differ between the UHs as a function of the CL position (9.46 ± 0.95 vs. 11 ± 0.95 pg/mL/mg; P > 0.05). The cellular stimulators promoted an increase in PGF2α synthesis (P < 0.02), and the effects differed among the animals (P < 0.04). The PGF2a production was higher in the explants treated with PDBu rather than OT (13.68 ± 1.16 vs. 10.01 ± 1.16 pg/mL/mg tissue, respectively; P < 0.05). In conclusion, PGF2α synthesis is modulated by the presence of the DF (local E2) but not the CL (local P4), and both PDBu and OT stimulated PGF2a synthesis.(AU)

A prostaglandina F2α (PGF2α) determina a luteólise em bovinos. A capacidade de manipular sua síntese endógena é indispensável para a produção animal em grande escala. O estradiol (E2) e a progesterona (P4) modulam diversas vias moleculares das células endometriais, incluindo a síntese de PGF2α; no entanto, pouco se sabe sobre seus mecanismos específicos. Este trabalho investigou a produção in vitro e a possível modulação da PGF2α endometrial devido a um efeito local do E2 endógeno no corno uterino ipsilateral ao folículo dominante (FD) ou da P4 no corno ipsilateral ao corpo lúteo (CL). Os estimuladores de PGF2α oxitocina (OT) e 12,23-dibutirato de forbol (PDBu) foram incubados com explantes endometriais, e o conteúdo de PGF2α foi mensurado. Para tal, vacas cíclicas foram sincronizadas, o desenvolvimento de FD e CL foi examinado por ultrassonografia, e no 17º dia do ciclo estral os explantes endometriais foram coletados e cultivados em meio ou suplementados com PDBu 10-6M ou 10-6M OT. As amostras de meio foram coletadas imediatamente após o tratamento e sessenta minutos depois. O radioimunoensaio mostrou que o conteúdo de PGF2α do corno ipsilateral ao FD foi 49% menor que o do corno contralateral (8,22 ± 0,95 vs. 12,24 ± 0,95 pg/mL/mg de tecido, respectivamente, P < 0,01). No entanto, os níveis de PGF2α não diferiram entre os cornos em função da posição do CL (9,46 ± 0,95 versus 11 ± 0,95 pg/mL/mg; P > 0,05). Os estimuladores celulares promoveram um aumento na síntese de PGF2α (P < 0,02), e os efeitos diferiram entre os animais (P < 0,04). A produção de PGF2α foi maior nos explantes tratados com PDBu em comparação à OT (13,68 ± 1,16 versus 10,01 ± 1,16 pg/mL/mg de tecido, respectivamente, P < 0,05). A conclusão obtida foi que a síntese de PGF2α é: modulada pela presença do FD (E2 local), mas não do CL (P4 local); e estimulada por PDBu e OT.(AU)

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Background: Ovarian cysts are commonly observed pathologies, which interfere with normal cyclic activity and adversely affect fertility in cows. Beta-carotene is effective in the reduction of reproductive problems by inducing the natural defence mechanisms of the body. There are several methods that can be used for the treatment of ovarian cysts. The separate and combined use of GnRH and PGF2α commonly uses in the treatment of ovarian cysts. Therefore, in the presented study the effects of Beta-carotene (βC) addition for the treatment of ovarian cysts either with GnRH solely or GnRH and PGF2α in combination on the fertility parameters of dairy cows were investigated.Materials, Methods & Results: Seventy-six Holstein Friesian cows having ovarian cysts diagnosed by ultrasonography (USG) were divided into three groups. Cows in Group I (GI, n = 27), were injected with GnRH (Buserelin acetate, 5 mL, im), PGF2α (Tiaprost-trometamol, 5 mL, im) and βC (20 mL/cow, into 4 regions by im route). In Group II (GII, n = 25) GnRH (Buserelin acetate, 5 mL, im) and PGF2α (Tiaprost-trometamol, 5mL, im) were administrated while GnRH (Buserelin acetate, 5 mL, im) solely in Group III (GIII, n = 24). Cysts were monitored via USG, and blood samples were collected on the on day of treatment (day 0) and on the 7th and 14th days following the administrations. Cows shoving oestrous were inseminated and pregnancy diagnoses were performed on the 40th day following insemination. Treatment results showed that there were statistically no significant differences between GI and GII (P > 0.05). Only numerical difference obtained in time from therapy to pregnancy and overall pregnancy index (P > 0.05). Overall pregnancy rate (85 %), first service pregnancy rates (40 %) and overall pregnancy index (2.11) in GI were found significantly higher than GIII (53.3 %; 20 %; 4.12) [P 0.05).[...]

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Background: Ovarian cysts are commonly observed pathologies, which interfere with normal cyclic activity and adversely affect fertility in cows. Beta-carotene is effective in the reduction of reproductive problems by inducing the natural defence mechanisms of the body. There are several methods that can be used for the treatment of ovarian cysts. The separate and combined use of GnRH and PGF2α commonly uses in the treatment of ovarian cysts. Therefore, in the presented study the effects of Beta-carotene (βC) addition for the treatment of ovarian cysts either with GnRH solely or GnRH and PGF2α in combination on the fertility parameters of dairy cows were investigated.Materials, Methods & Results: Seventy-six Holstein Friesian cows having ovarian cysts diagnosed by ultrasonography (USG) were divided into three groups. Cows in Group I (GI, n = 27), were injected with GnRH (Buserelin acetate, 5 mL, im), PGF2α (Tiaprost-trometamol, 5 mL, im) and βC (20 mL/cow, into 4 regions by im route). In Group II (GII, n = 25) GnRH (Buserelin acetate, 5 mL, im) and PGF2α (Tiaprost-trometamol, 5mL, im) were administrated while GnRH (Buserelin acetate, 5 mL, im) solely in Group III (GIII, n = 24). Cysts were monitored via USG, and blood samples were collected on the on day of treatment (day 0) and on the 7th and 14th days following the administrations. Cows shoving oestrous were inseminated and pregnancy diagnoses were performed on the 40th day following insemination. Treatment results showed that there were statistically no significant differences between GI and GII (P > 0.05). Only numerical difference obtained in time from therapy to pregnancy and overall pregnancy index (P > 0.05). Overall pregnancy rate (85 %), first service pregnancy rates (40 %) and overall pregnancy index (2.11) in GI were found significantly higher than GIII (53.3 %; 20 %; 4.12) [P < 0.05]. No significant difference was observed in progesterone (P4) levels between the groups (P > 0.05).[...](AU)

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Seventy six ewes were treated with 7.5, 12.5, 25 or 50µg of cloprostenol on day 6 or 9 post-estrus to compare the luteolytic efficiency of the PGF2α analogue at each stage and to evaluate if progesterone concentrations at the time of treatment affect such efficiency. Blood samples were obtained before cloprostenol administration and 12, 24, 48, and 72h thereafter. There was an effect of dose (p<0.05) but not of day post-estrus on the proportion of animals completing luteolysis. As the dose increased, the proportion of ewes completing luteolysis also increased. Also, as the dose increased from 7.5 to 25µg, more ewes showed a transient progesterone decline instead of an absence of response, indicating that in some ewes reduced doses initiated luteolysis but were not able to finish the process. Since the dose of 25µg resulted in close to 50% luteolytic efficacy, this group was used to study the effects of progesterone concentrations at the time of treatment on the response to cloprostenol. Pre-treatment progesterone concentrations were higher (p<0.01) in ewes experiencing luteolytic failure than in those that completed luteolysis. There was a negative correlation between initial progesterone concentrations and their reduction by 12h post-treatment. It is concluded that high progesterone concentrations are associated with a reduction in sensitivity to small doses of cloprostenol. Possible mechanisms and implications of this luteoprotective effect are discussed.

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This study evaluated the efficiency of two d-cloprostenol injections at different intervals on the reproductive parameters of dairy goats. Trial 1 comprised 54 goats allocated to receive two 37.5µg d-cloprostenol doses at intervals of seven (T7, n=19), 10 (T10, n=18), and 11.5 (T11.5, n=17) days. Trial 2 comprised 62 goats allocated to receive injections at T7 (n=30) and T11.5 (n=32). Ultrasonography was done and blood was collected just before d-cloprostenol injections. After the second dose, goats were artificially inseminated (AI) with frozen-thawed semen at 18-24h (Trial 1) or at 10-24h (adjusted according to the time of estrus onset in Trial 2) after estrus detection. Estrus response rate did not differ (P>0.05) among groups in Trials 1 (T7=94.7%; T10=88.9%; T11.5=88.2%) and 2 (T7=90.0%; T11.5=96.9). All females showed progesterone concentrations >1ng/mL before both d-cloprostenol injections. The largest follicle diameter present on ovaries was similar (P>0.05) among treatments at the first and second dose. The second largest follicle diameter was superior (P<0.05) to T7 than to T10 and T11.5 goats at first dose only. This possibly resulted in lower interval to estrus (P<0.05) in T7-treated goats than other treated goats in both trials. The conception rate was similar among treatment groups in Trials 1 (T7=55.6%; T10=18.8%; T11.5=26.7%) and 2 (T7=85.2%; T11.5=93.6%). The three treatments efficiently synchronized estrus. T7 and T11.5 protocols resulted in high estrus synchrony and conception rates when adjusting the AI time according to interval of estrus.

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STUDY QUESTION: What is the role of the endocannabinoid system (eCS) in the alterations of the endocrine system in a murine model of lipopolysaccharide (LPS)-induced miscarriage? SUMMARY ANSWER: In 7-days pregnant wild type, but not cannabinoid receptor type 1 knockout (CB1-KO) mice, LPS increased COX-2 expression and prostaglandin F2α (PGF2α) production in the uterus leading to lower expression of prolactin receptor in the ovary and a marked regression of corpora lutea (CL), suggesting that the eCS mediates the deleterious effects of LPS on reproductive events. WHAT IS KNOWN ALREADY: Appropriate systemic progesterone levels are critical for a successful pregnancy outcome. Precocious loss of luteal progesterone (P4) secretion leads to miscarriage in rodents. We have previously shown that LPS administration to pregnant mice induces embryonic resorption accompanied by a dramatic decrease in systemic progesterone levels in a murine model of inflammatory miscarriage, with the eCS mediating these LPS-induced deleterious effects. STUDY DESIGN SAMPLES/MATERIALS, METHODS: CD1 wild-type (WT) and CB1-KO mice were randomly allocated to Vehicle (saline; i.p.) or LPS (0.5 µg/g body weight; i.p.) treated groups: (WT-Vehicle; WT-LPS; CB1-KO-Vehicle and CB1-KO-LPS). A single injection was given on day 7 of pregnancy and tissues (blood, ovary, uterus) were collected 6, 12, 24 and 48 h later. P4 and PGF2α plasma levels were determined by radioimmunoassay. Cyclooxygenase-2 (COX-2) mRNA (RT-PCR) and protein (Western blot) content in uterus was assayed. COX-2 and prolactin receptor (PrlR) mRNA levels in the ovary were assayed by RT-PCR. Tissue morphology of the CL was assessed by haematoxylin-eosin staining. MAIN RESULTS AND THE ROLE OF CHANCE: Treatment of 7-day pregnant WT mice with LPS induced a P4 withdrawal (p < 0.05), increased in uterine COX-2 mRNA and protein expression (p < 0.05) as well as an increase in uterine PGF2α production (p < 0.05). These changes were absent in LPS-treated 7-day pregnant CB1-KO mice. In ovarian tissues, LPS treatment to 7-day pregnant WT mice induced a downregulation of PrlR mRNA expression (p < 0.05) together with an increase in COX-2 mRNA expression (p < 0.05) and PGF2α content (p < 0.05). These effects were absent in the CB1-KO mice. Collectively, our results suggest a role for the eCS mediating LPS-induced deleterious effects on reproductive tissues. LIMITATIONS, REASONS FOR CAUTION: An important caveat of this study is the endocrine differences between mice and humans during pregnancy (e.g. P4 is produced by the CL throughout pregnancy in mice, whereas this is not the case in humans), which limits the extrapolation of the results presented here. WIDER IMPLICATIONS OF THE FINDINGS: Our findings provide new insights in the role of the endocannabinoid system in the physiopathology of reproduction as well as the role of this endogenous system as a mediator of LPS deleterious effects on reproductive tissues. LARGE SCALE DATA: None. STUDY FUNDING AND COMPETING INTERESTS: Dr Ana María Franchi was funded by Agencia Nacional para la Promoción Científica y Tecnológica (PICT 2010/0813 and PICT 2013/0097) and by Consejo Nacional de Investigaciones Científicas y Técnicas (PIP 2012/0061). The authors have no competing interests.

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The aim of this study was to evaluate luteolysis using three doses of PGF2α on Day 5 or Day 7 of the estrous cycle in nonlactating Nellore (Bos indicus) cows. Cows (n = 323) were assigned within date of estrus (Day 0 of estrous cycle) to receive 12.5, 25.0, or 50.0 mg of PGF2α on either Day 5 or Day 7 of the estrous cycle in a 3 × 2 factorial arrangement. Blood samples for progesterone (P4) concentrations were collected at 0, 24, 48, and 72 hours after PGF2α to assess luteolysis (L). Luteolysis was defined on the basis of P4 concentrations at 72 hours using either less than 0.5 ng/mL (L0.5) or less than 1.0 ng/mL (L1.0) as the cut off. Luteolysis was considered "partial" when P4 concentration declined within 24 hours after PGF2α but failed to decline further or, in some cases, increased. Incidence of luteolysis was less (P < 0.01) on Day 5 than Day 7 of the estrous cycle (17.3 vs. 47.6% and 30.4 vs. 77.2%; for L0.5 and L1.0, respectively). Dose of PGF2α increased (P < 0.01) L1.0 (12.5 mg = 38.9%; 25.0 mg = 52.3%; and 50.0 mg = 70.4%). Incidence of partial luteolysis for cows on Day 5 (57.1%) was greater (P < 0.01) than that on Day 7 (19.1%) of the estrous cycle and was more prevalent (P < 0.01) with lower doses of PGF2α (12.5 mg = 49.1%; 25.0 mg = 37.4%; and 50.0 mg = 27.8%). In conclusion, both days of the estrous cycle and doses of PGF2α influenced the incidence of complete and partial luteolysis in Nellore cows and should be an important consideration when devising estrus synchronization programs in this species.

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