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Background: Identifying molecular mechanisms responsible for the response to heat stress is essential to increase production, reproduction, health, and welfare. This study aimed to identify early biological responses and potential biomarkers involved in the response to heat stress and animal's recovery in tropically adapted beef cattle through proteomic analysis of blood plasma. Methods: Blood samples were collected from 14 Caracu males during the heat stress peak (HSP) and 16 h after it (heat stress recovery-HSR) assessed based on wet bulb globe temperature index and rectal temperature. Proteome was investigated by liquid chromatography-tandem mass spectrometry from plasma samples, and the differentially regulated proteins were evaluated by functional enrichment analysis using DAVID tool. The protein-protein interaction network was evaluated by STRING tool. Results: A total of 1,550 proteins were detected in both time points, of which 84 and 65 were downregulated and upregulated during HSR, respectively. Among the differentially regulated proteins with the highest absolute log-fold change values, those encoded by the GABBR1, EPHA2, DUSP5, MUC2, DGCR8, MAP2K7, ADRA1A, CXADR, TOPBP1, and NEB genes were highlighted as potential biomarkers because of their roles in response to heat stress. The functional enrichment analysis revealed that 65 Gene Ontology terms and 34 pathways were significant (P < 0.05). We highlighted those that could be associated with the response to heat stress, such as those related to the immune system, complement system, hemostasis, calcium, ECM-receptor interaction, and PI3K-Akt and MAPK signaling pathways. In addition, the protein-protein interaction network analysis revealed several complement and coagulation proteins and acute-phase proteins as important nodes based on their centrality and edges. Conclusion: Identifying differentially regulated proteins and their relationship, as well as their roles in key pathways contribute to improve the knowledge of the mechanisms behind the response to heat stress in naturally adapted cattle breeds. In addition, proteins highlighted herein are potential biomarkers involved in the early response and recovery from heat stress in tropically adapted beef cattle.
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This study aimed to evaluate the performance, feed efficiency, feeding behavior, and cortisol concentrations of lambs ranked into different groups of Residual Feed Intake (RFI) and Residual Intake and Gain (RIG). Forty Santa Inês lambs with a mean initial age of 120 ± 30 days and mean initial (IBW) and final body weight (FBW) of 28.9 ± 3.2 and 43.7 ± 3.9 kg, respectively, were used. The animals were fed at an automatic feed and water station (Intergado®) for 60 days and classified at the end of the experiment as low-RFI/RIG, medium-RFI/RIG, and high-RFI/RIG according to RFI and RIG. Performance, feed efficiency measures, ingestive behavior, temperament, and cortisol concentrations were measured, and Pearson's correlation analysis was performed to evaluate the relationship between the variables studied. The variables that differed most between RFI groups were dry matter intake, feed conversion, and feed efficiency, with better values in efficient animals (RFI-). Regarding the RGI groups, the average daily gain of animals classified as RIG- was 19.37% lower than that of animals classified as RIG± and 39.28% lower than that of animals classified as RIG+. Only the hourly feeding rate per day (p = 0.04) differed significantly between RFI and RIG classes, while no differences were observed for the ingestive behavior variables or cortisol. The number of visits to the feed bunk (total, with or without intake) showed no correlation with any performance or efficiency measure. Identifying animals ranked into different RFI and RIG classes shows that more efficient animals have a lower feed intake for the same weight gain and spend less time at the feed bunk.
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Hidrocortisona , Carneiro Doméstico , Ovinos , Animais , Peso Corporal , Ingestão de Alimentos , Comportamento AlimentarRESUMO
Menstrual blood mesenchymal stem cells (MenSCs) have gained prominence in the endometriosis scientific community, given their multifunctional roles in regenerative medicine as a noninvasive source for future clinical applications. In addition, changes in post-transcriptional regulation via miRNAs have been explored in endometriotic MenSCs with a role in modulating proliferation, angiogenesis, differentiation, stemness, self-renewal, and the mesenchymal-epithelial transition process. In this sense, homeostasis of the miRNA biosynthesis pathway is essential for several cellular processes and is related to the self-renewal and differentiation of progenitor cells. However, no studies have investigated the miRNA biogenesis pathway in endometriotic MenSCs. In this study, we profiled the expression of eight central genes for the miRNA biosynthesis pathway under experimental conditions involving a two-dimensional culture of MenSCs obtained from healthy women (n = 10) and women with endometriosis (n = 10) using RT-qPCR and reported a two-fold decrease in DROSHA expression in the disease. In addition, miR-128-3p, miR-27a-3p, miR-27b-3p, miR-181a-5p, miR-181b-5p, miR-452-3p, miR-216a-5p, miR-216b-5p, and miR-93-5p, which have been associated with endometriosis, were identified through in silico analyses as negative regulators of DROSHA. Because DROSHA is essential for miRNA maturation, our findings may justify the identification of different profiles of miRNAs with DROSHA-dependent biogenesis in endometriosis.
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Endometriose , Células-Tronco Mesenquimais , MicroRNAs , Humanos , Feminino , Regulação para Baixo/genética , Endometriose/genética , Endometriose/metabolismo , MicroRNAs/metabolismo , Regulação da Expressão Gênica , Células-Tronco Mesenquimais/metabolismo , Ribonuclease III/genética , Ribonuclease III/metabolismoRESUMO
The objective was to evaluate the performance, carcass and meat quality of 40 lambs classified by RFI (residual feed intake) and RIG (residual intake and gain). Dry matter intake (DMI) was recorded to calculate the RFI and RIG, classified as efficient, moderately or inefficient. After the confinement period, they were slaughtered and the carcass and meat quality were determined. The efficient animals had DMI scores of 0.700 RFI and 0.400 kg/d RIG, lower than the inefficient ones with similar weight gain. The RFI efficient animals showed greater shear force, without effect in the RIG classification. In general, the variables analyzed were not influenced by RFI or RIG. Efficiency measures do not affect the carcass and meat quality of sheep, but they do have the advantage of identifying animals with lower feed consumption, making the system more efficient. However, as the dataset is limited to fully assess the effects, this manuscript can be used as preliminary results for future studies.
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Ração Animal , Ingestão de Alimentos , Animais , Ovinos , Ração Animal/análise , Fenótipo , Aumento de Peso , Carne/análiseRESUMO
Climate change is a reality and global surface temperature is projected to rise substantially in the next 80 years. Agriculture practices will have to adapt to climate change, and also help to mitigate this effect using, among other strategies, forest conservation and management. Silvopastoral systems have been adopted in tropical climate livestock areas but their benefits on thermal comfort and reproductive performance of beef cows are not completely known. Therefore, our aims were to compare the microclimate of silvopastoral and intensive rotational unshaded grazing systems in different months and to evaluate physiological variables (Exp. 1 and 2), metabolism, and in vitro embryo production (Exp. 2) in crossbred beef females. Our hypothesis is that the silvopastoral system can improve the thermal comfort of beef heifers and cows and, consequently, also improve dry matter intake, body weight gain, and in vitro embryo production when compared to the unshaded rotational grazing system. In Exp 1, the silvopastoral system decreased body temperature and increased welfare and performance of heifers. In Exp. 2, the silvopastoral system enhanced the body weight but did not affect metabolism and the general reproductive performance, but increased the recovery rate of oocytes in primiparous cows.
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The objective of this study was to evaluate the influence of thermal stress indices and minimum temperature on the semen quality of dairy Gyr bulls and to determine whether the minimum night temperature is able to balance the thermal stress that occurred during the day. A total of 922 semen samples were collected from 391 bulls aged between 15 and 49 months, which participated in progeny pretest trials, from December 2011 to April 2017. Semen characteristics (ejaculate volume, gross motility, sperm motility, sperm vigor, sperm concentration, and percentage of total defects) and scrotal circumference were analyzed. Meteorological data were recorded daily by the National Institute of Meteorology. The analyses were carried out using mixed models. The following fixed effects were included in the model: test, month of sample collection, scrotal circumference, the age of the bull at sampling as covariate (linear regression), and the climatic variables (equivalent temperature index [ETI], temperature and humidity index [THI], or minimum temperature). The effect of the bull nested within the test was included in the model as a random effect. Two periods were considered to evaluate the effect of the climatic variables on sperm quality: 7 days before collection (period 1) and the day of collection (period 2). There was an effect of ETI on sperm vigor and concentration in both periods and on volume and percentage of total defects only in the second period. The THI exerted a significant effect on volume in both periods and on sperm motility and concentration only in the second period. There was an effect of the minimum temperature on ejaculate volume, sperm motility, sperm concentration, and percentage of total defects only in the second period, which corresponds to the day of semen collection. Taken together, the results show that there was sufficient thermal stress to negatively affect semen quality. However, the minimum temperature during the night was a significant factor that balanced these negative effects of THI and ETI on semen traits of Gyr bulls.
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Análise do Sêmen , Motilidade dos Espermatozoides , Animais , Bovinos , Pré-Escolar , Humanos , Lactente , Masculino , Sêmen , Contagem de Espermatozoides , EspermatozoidesRESUMO
OBJECTIVE: The aim of the present study was to analyze the expression of the CD63, S100A6, and GNB2L1genes, which participate in mechanisms related to the complex pathophysiology of endometriosis. METHODS: A case-control study was conducted with 40 women who were diagnosed with endometriosis, and 15 fertile and healthy women. Paired samples of eutopic endometrium and endometriotic lesions (peritoneal and ovarian endometriotic implants) were obtained from the women with endometriosis in the proliferative (n = 20) or secretory phases (n = 20) of the menstrual cycle. As controls, paired endometrial biopsy samples were collected from the healthy women in the proliferative (n = 15) and secretory (n = 15) phases of the same menstrual cycle. We analyzed the expression levels of the CD63, S100A6, and GNB2L1 genes by real-time polymerase chain reaction. RESULTS: An increase in CD63, S100A6, and GNB2L1 gene transcript levels was observed in the ectopic implants compared with the eutopic endometrium of the women with and without endometriosis, regardless of the phase of the menstrual cycle. CONCLUSION: These findings suggest that the CD63, S100A6, and GNB2L1 genes may be involved in the pathogenesis of endometriosis, since they participate in mechanisms such as inhibition of apoptosis, angiogenesis and cell proliferation, which lead to the loss of cell homeostasis in the ectopic endometrium, thus contributing to the implantation and survival of the tissue in the extrauterine environment.
OBJETIVO: O objetivo do presente estudo foi analisar a expressão dos genes CD63, S100A6 e GNB2L1, que participam em mecanismos relacionados à complexa fisiopatologia da endometriose. MéTODOS: Um estudo caso-controle foi realizado com 40 mulheres diagnosticadas com endometriose e 15 mulheres férteis e saudáveis. Amostras pareadas de endométrio eutópico e de lesões endometrióticas (implantes endometrióticos peritoneais e ovarianos) foram obtidas de mulheres com endometriose nas fases proliferativa (n = 20) ou secretora (n = 20) do ciclo menstrual. Como controle, amostras pareadas de biópsia endometrial foram coletadas de mulheres saudáveis nas fases proliferativa (n = 15) e secretora (n = 15) no mesmo ciclo menstrual. Foram analisados os níveis de expressão dos genes CD63, S100A6 e GNB2L1 por reação em cadeia da polimerase em tempo real. RESULTADOS: Foi observado um aumento nos níveis de transcritos dos genes CD63, S100A6 e GNB2L1 em implantes ectópicos quando comparado ao endométrio eutópico de mulheres com e sem endometriose, independente da fase do ciclo menstrual. CONCLUSãO: Estes achados sugerem que os genes CD63, S100A6 e GNB2L1 podem estar envolvidos na patogênese da endometriose, pois participam de mecanismos como inibição de apoptose, angiogênese e proliferação celular, os quais levam à perda da homeostase celular no endométrio ectópico e, portanto, contribuem para o implante e a sobrevivência do tecido no ambiente extrauterino.
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Apoptose/genética , Proteínas de Ciclo Celular/genética , Proliferação de Células/genética , Endometriose/genética , Endometriose/patologia , Proteínas de Neoplasias/genética , Neovascularização Patológica/genética , Receptores de Quinase C Ativada/genética , Proteína A6 Ligante de Cálcio S100/genética , Tetraspanina 30/genética , Adulto , Estudos de Casos e Controles , Feminino , Expressão Gênica , HumanosRESUMO
Semen cryopreservation comprises different steps, among them are the cooling and freezing rates which significantly influence the quality of thawed sperm. Different systems with variable freezing rates are used for freezing bull semen in the field, with a consequence of variable success rates. The objective of this study was to compare different systems for freezing bull semen in the field. Five cooling methods of semen and two methods for the subsequent freezing phase (5â¯×â¯2 factorial scheme) were used. Two to four ejaculates were collected from 12 bulls with an electroejaculator. The ejaculates were diluted in BotuBov® to a concentration of 50â¯×â¯106 spermatozoa/mL in 0.5-mL straws. After dilution, the straws were cooled to 5⯰C in five cooling systems: TK 4000® at a cooling rate of -0.25⯰C/min (R1); TK 4000® at a rate of -0.5⯰C/min (R2); Minitube® refrigerator at a rate of -2.8⯰C/min (R3); Botutainer® at a rate of -0.65⯰C (R4), and domestic refrigerator at a rate of -2.0⯰C/min (R5). After stabilization at 5⯰C for 4â¯h, these straws were then submitted to two freezing systems: TK 4000® at a freezing rate of -15⯰C/min (C1) and Styrofoam box with liquid nitrogen at a rate of -19⯰C/min (C2). Sperm kinetics were evaluated by computer-assisted sperm analysis at four time points: in fresh semen, after cooling, post-thawing, and after the rapid thermal resistance test (TRT). In addition, plasma and acrosomal membrane integrity, mitochondrial potential and intracellular H2O2 were analyzed after thawing by flow cytometry. The R1, R2 and R4 cooling systems were the most efficient in preserving sperm viability, membrane integrity and intracellular H2O2. Samples frozen in the C1 system exhibited better post-thaw and post-TRT kinetics than C2 samples. In conclusion, slower cooling curves in conjunction with a constant freezing rate obtained with the programmable unit were more efficient for freezing bull semen in the field.
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Bovinos/fisiologia , Criopreservação/veterinária , Congelamento , Preservação do Sêmen/veterinária , Sêmen/fisiologia , Animais , Masculino , Fatores de TempoRESUMO
Inadequate corral facilities and improper handling are major causes of stress in beef cattle. The purpose of this study was to evaluate the effects of minor changes in the corral and adoption of good handling practices on the behavior, cortisol release, and time spent taking blood samples in Nellore cows. Minor corral changes included obstructing the cow's vision when the handler walked deep into the animal's flight zone and the elimination of bright objects, color contrasts, puddles, shadows, and darkness in the corral. Handling was improved by eliminating dogs, electric goads (prods), and yelling, as well as adopting a calm behavior. A total of 141 Nellore cows from two typical extensive livestock farms were studied. The cows were evaluated individually before and after the corral changes. Blood samples were collected in the restraint device for cortisol measurement. The minor corral changes and the adoption of good handling practices result in better results for all variables studied. The results showed differences in the interactions between treatment and ranch for chute score (P = 0.0091) and exit score (P < 0.0001). The cortisol release was lower (P < 0.001) and better for the improved methods, resulting in calmer cows compared to cortisol released before the minor changes (41.03 ± 2.9 vs 60.40 ± 3.8 ng/mL). Minor changes made in the corrals and the adoption of good handling practices were effective in improving cow behavior in the chute and in reducing exit velocity, cortisol released, and the time spent taking blood samples.
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Criação de Animais Domésticos/métodos , Comportamento Animal , Abrigo para Animais , Hidrocortisona/sangue , Bem-Estar do Animal , Animais , Bovinos , Cães , Estresse Psicológico/prevenção & controleRESUMO
Some studies have demonstrated alterations in the composition of peritoneal fluid (PF) from women with endometriosis. Controversial studies have suggested that impaired oocyte quality may be involved in the pathogenesis of endometriosis-related infertility. The aim of this study was to evaluate the spindle and chromosome distribution of in vitro-matured oocytes in the presence of 2 concentrations of PF from infertile women with minimal/mild endometriosis (EI/II) compared to fertile controls. We performed an experimental study using a bovine model. Samples of PF were obtained from 12 women who underwent videolaparoscopy-6 infertile women with EI/II and 6 fertile women without endometriosis (control group). Immature bovine oocytes underwent in vitro maturation (IVM) in the absence of PF and in the presence of 2 concentrations (1% and 10%) of PF from fertile women and from infertile women with EI/II. After 22 to 24 hours of IVM, oocytes were fixed for subsequent immunofluorescence staining for the visualization of microtubules and chromosomes by confocal microscopy. The percentage of meiotically normal oocytes was significantly lower for oocytes that underwent IVM in the presence of 1% (62.50%) and 10% (56.25%) of PF from infertile women with EI/II than in the absence of PF (88.46%) and in the presence of 1% (78.57%) and 10% (84.61%) of PF from fertile women ( P < .01). We demonstrated that PF from infertile women with EI/II promotes meiotic abnormalities in in vitro-matured bovine oocytes. Therefore, our results contribute to the understanding of the etiopathogenic mechanisms of infertility related to EI/II.
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Líquido Ascítico/patologia , Endometriose/patologia , Infertilidade Feminina/patologia , Oócitos/patologia , Fuso Acromático/patologia , Animais , Bovinos , Feminino , Humanos , Técnicas de Maturação in Vitro de Oócitos , Metáfase , Microscopia Confocal , Projetos PilotoRESUMO
The aim of this study was to estimate genetic parameters for accumulated productivity (ACP), first calving interval (CI1), second calving interval (CI2), and mean calving interval (MCI) in Nelore beef cattle. The ACP trait is a reproduction index and comprises the total number of calves born per dam, weight of weaned calves, and age of the dam at calving. Genetic parameters were estimated by the average information restricted maximum likelihood method in two-trait analyses. The average heritability estimate for ACP was 0.17 (0.03). For CI1, CI2, and MCI, the heritability estimates were 0.02, 0.02, and 0.06, respectively. Genetic correlations between ACP with CI1, CI2, and MCI were -0.16 ± 0.47, -0.29 ± 0.53, and -0.40 ± 0.27, respectively. Despite of the low heritability estimates obtained in our study, reproduction traits should be further studied and their inclusion in the selection criteria must be evaluated in order to improve the performance of females because these traits are of great economic importance in beef cattle. Accumulated productivity could contribute to decrease the mean calving interval in Nelore cattle.
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Bovinos/fisiologia , Reprodução/fisiologia , Animais , Brasil , Bovinos/genética , Feminino , Funções Verossimilhança , Carne , Fenótipo , Reprodução/genéticaRESUMO
It is questioned whether worsening of oocyte quality and oxidative stress (OS) are involved in the pathogenesis of infertility related to endometriosis and in compromised intracytoplasmic sperm injection (ICSI) outcomes. Cumulus cells (CCs) protect oocytes from entering apoptosis induced by OS. Thus, we carried out a case-control study comparing expression of superoxide dismutase 1 (SOD1), superoxide dismutase 2 (SOD2), and glutathione peroxidase 4 (GPX4; genes encoding for the main antioxidant enzymes) in CCs from mature oocytes of 26 infertile patients with minimal/mild endometriosis, 14 patients with moderate/severe endometriosis, and 41 controls undergoing controlled ovarian stimulation for ICSI, using real-time polymerase chain reaction. As a secondary objective, we investigated the interaction between the expression of these genes and clinical pregnancy (CP) by a statistical model. Only infertile women with moderate/severe endometriosis showed increased expression of the SOD1 in CCs compared to women with minimal/mild endometriosis and controls, with a positive interaction between increased expression and the occurrence of CP, suggesting that SOD1 might be a potential biomarker of CP following ICSI.
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Células do Cúmulo/enzimologia , Endometriose/complicações , Infertilidade Feminina/etiologia , Superóxido Dismutase/genética , Adulto , Estudos de Casos e Controles , Endometriose/diagnóstico , Feminino , Humanos , Infertilidade Feminina/diagnóstico , Infertilidade Feminina/enzimologia , Infertilidade Feminina/genética , Infertilidade Feminina/terapia , Estudos Prospectivos , Reação em Cadeia da Polimerase em Tempo Real , Técnicas de Reprodução Assistida , Índice de Gravidade de Doença , Superóxido Dismutase-1 , Regulação para CimaRESUMO
Aromatase plays a fundamental role in the establishment of oocyte quality, which might be compromised in infertile women with endometriosis. The expression of the CYP19A1 gene (that encodes aromatase) was compared in cumulus cells and oestradiol concentrations in the follicular fluid of infertile women with and without endometriosis submitted to ovarian stimulation for intracytoplasmic sperm injection. Cumulus cells were isolated and the expression of the CYP19A1 was quantitated through real-time polymerase chain reaction. Oestradiol concentrations in follicular fluid were measured by chemiluminescence immunoassay. A lower expression of the CYP19A1 in the cumulus cells of infertile women with endometriosis was observed compared with controls (0.17 ± 0.13 and 0.56 ± 0.12, respectively), and no significant difference in the follicular fluid oestradiol concentrations was observed between groups. Our results show reduced expression of the CYP19A1 in cumulus cells of infertile women with endometriosis, which may play a role in the pathogenesis of endometriosis-related infertility.
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Aromatase/genética , Células do Cúmulo/enzimologia , Regulação para Baixo , Endometriose/genética , Infertilidade Feminina/genética , Estudos de Casos e Controles , Endometriose/complicações , Feminino , Humanos , Infertilidade Feminina/complicações , Indução da Ovulação , Gravidez , Taxa de Gravidez , Estudos Prospectivos , Injeções de Esperma IntracitoplásmicasRESUMO
OBJECTIVE: To elucidate the potential mechanisms involved in the physiopathology of endometriosis. We analyzed the differential gene expression profiles of eutopic and ectopic tissues from women with endometriosis. DESIGN: Prospective laboratory study. SETTING: University hospital. PATIENT(S): Seventeen patients in whom endometriosis was diagnosed and 11 healthy fertile women. INTERVENTION(S): Endometrial biopsy specimens from the endometrium of healthy women without endometriosis and from the eutopic and ectopic endometrium tissues of patients with endometriosis were obtained in the early proliferative phase of the menstrual cycle. MAIN OUTCOME MEASURE(S): Six paired samples of eutopic and ectopic tissue were analyzed by subtractive hybridization. To evaluate the expression of genes found by rapid subtraction hybridization methods, we measured CTGF, SPARC, MYC, MMP, and IGFBP1 genes by real-time polymerase chain reaction in all samples. RESULT(S): This study identified 291 deregulated genes in the endometriotic lesions. Significant expression differences were obtained for SPARC, MYC, and IGFBP1 in the peritoneal lesions and for MMP3 in the ovarian endometriomas. Additionally, significant differences were obtained for SPARC and IGFBP1 between the peritoneal and ovarian lesions. No significant differences were found for the studied genes between the control and the eutopic endometrium. CONCLUSION(S): This study identified 291 genes with differential expression in endometriotic lesions. The deregulation of the SPARC, MYC, MMP3, and IGFBPI genes may be responsible for the loss of cellular homeostasis in endometriotic lesions.