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Abstract Senescent cells have been linked to the pathogenesis of metabolic dysfunction-associated steatotic liver disease (MASLD). However, the effectiveness of senolytic drugs in reducing liver damage in mice with MASLD is not clear. Additionally, MASLD has been reported to adversely affect male reproductive function. Therefore, this study aimed to evaluate the protective effect of senolytic drugs on liver damage and fertility in male mice with MASLD. Three-month-old male mice were fed a standard diet (SD) or a choline-deficient western diet (WD) until 9 months of age. At 6 months of age mice were randomized within dietary treatment groups into senolytic (dasatinib + quercetin [D + Q]; fisetin [FIS]) or vehicle control treatment groups. We found that mice fed choline-deficient WD had liver damage characteristic of MASLD, with increased liver size, triglycerides accumulation, fibrosis, along increased liver cellular senescence and liver and systemic inflammation. Senolytics were not able to reduce liver damage, senescence and systemic inflammation, suggesting limited efficacy in controlling WD-induced liver damage. Sperm quality and fertility remained unchanged in mice developing MASLD or receiving senolytics. Our data suggest that liver damage and senescence in mice developing MASLD is not reversible by the use of senolytics. Additionally, neither MASLD nor senolytics affected fertility in male mice.

Ames dwarf mice (df/df) display delayed aging relative to their normal (N) siblings, living approximately 40–60 % longer. As such, investigating the mechanisms that enable these organisms to have extended lifespan is useful for the development of interventions to slow aging and deter age-related disease. Nonalcoholic fatty liver disease (NAFLD) is a condition that is characterized by the accumulation of excess adipose tissue in the liver. Previous studies highlight the potential of calorie restriction (CR) in promoting longevity, but little is known about its effects on the biomolecular processes that govern NAFLD. In this study, we examined the role of 6-month CR on genes regulating lipid metabolism in the livers of long-living df/df mice and their N littermates. Importantly, our findings showed significant downregulation of miR-34a-5p in N-CR mice and df/df mice regardless of dietary regimen. Alongside, our RT-PCR results indicated that downregulation of miR-34a-5p is correlated with the expression of metabolism-associated mRNAs involved in modulating the processes of de novo lipogenesis (DNL), fatty acid oxidation (FAO), very-low density lipoprotein transport (VLDL-T), and reverse cholesterol transport (RCT). To further verify the role of miR-34a-5p in regulating metabolic processes, we transfected the human liver cancer (HepG2) cell line with miR-34a mimic, and studied its effect on direct targets Sirt1, Ampk, and Ppara as well as downstream lipid transport regulating genes. Our findings suggest that CR and df/df life extending mutation are robust drivers of the miR-34a-5p signaling pathway and prevent the pathogenesis of age-related diseases by improving overall lipid homeostasis.

ABSTRACT Objective: To study associations between polymorphisms in the angiotensin converting enzyme (ACE I/D), actinin 3 (ACTN3 R577X) and paraoxonase 1 (PON1 T(-107)C) genes and chronic diseases (diabetes and hypertension) in women. Materials and methods: Genomic DNA was extracted from saliva samples of 78 women between 18 and 59 years old used for genetic polymorphism screening. Biochemical data were collected from the medical records in Basic Health Units from Southern Brazil. Questionnaires about food consumption, physical activity level and socioeconomic status were applied. Results: The XX genotype of ACTN3 was associated with low HDL levels and high triglycerides, total cholesterol and glucose levels. Additionally, high triglycerides and LDL levels were observed in carriers of the TT genotype of PON1, and lower total cholesterol levels were associated to the CC genotype. As expected, women with diabetes/hypertense had increased body weight, BMI (p = 0.02), waist circumference (p = 0.01), body fat percentage, blood pressure (p = 0.02), cholesterol, triglycerides (p = 0.02), and blood glucose (p = 0.01), when compared to the control group. Conclusion: Both ACTN3 R577X and PON1 T(-107)C polymorphisms are associated with nutritional status and blood glucose and lipid levels in women with diabetes/hypertense. These results contribute to genetic knowledge about predisposition to obesity-related diseases.

ABSTRACT Objective: The aim of this study was to evaluate the serum activity of PON1 in women according to SNPs L55M and T-107C and diet composition. Materials and methods: Blood and serum samples from 26 women were used. DNA extraction, PCR and digestion with restriction enzymes of the PCR fragment were performed for genotyping the PON1 SNPs T-107C and L55M. Serum PON1 activity was measured in a single time point. Patients completed the semi-quantitative food frequency questionnaire and diet composition was estimated. Results: Genotypic distribution for L55M SNP was 56% for the LL genotype, 32% for LM and 12% for MM; for the PON1 C(-107)T SNP it was 28% for the TT genotype, 41% for CT and 31% for CC. Individuals with C and L alleles had higher serum PON1 activity. Combining the two SNPs, we observed that individuals carrying the LL and CC genotypes had twice the activity of carriers of the TT and MM genotypes. Considering food intake, no significant difference was observed between genotypes and intake levels. Conclusion: PON1 T(-107)C and L55M SNPs exert a strong effect on serum PON1 activity in an additive manner and are more important than diet to predict serum PON1 activity.

ABSTRACT Objectives: Paraoxonase 1 (PON1) is an enzyme that has antioxidant potential, which confers a protective effect against the atherosclerotic process. However, studies associating genetics, dietary patterns and PON1 activity in individuals with cardiovascular disease (CVD) are scarce. Thus, the aim of the current study was to evaluate the influence of dietary factors on serum PON1 in CVD patients. Subjects and methods: Cross-sectional, sub-study of the BALANCE Program Trial. All patients aged 45 years or older and had evidence of established atherosclerotic disease in the preceding 10 years. Body weight, height, waist circumference, blood pressure, lipid profile and fasting glucose were collected. Food intake was assessed with 24-h dietary recall. Data was analyzed using SAS University Edition and a P value ≤ 0.05 was considered statistically significant. Sample was divided into three groups, according to the PON1 T(-107)C genotype (CC, CT and TT) and serum PON1 activity (Low, Medium, High). Results: There were no genotype differences for major factors. However, the systolic blood pressure was lower for CT individuals (p

Objective: The cardioprotective enzyme paraoxonase‐1 (PON1) suffers an important influence from genetic polymorphisms and nutritional factors. The aim of this study was to investigate the influence of diet, nutritional status, and the C(‐107)T polymorphism on PON1 arylesterase activity in children. Methods: This was a cross‐sectional study with 97 children, aged between 5 and 8 years, of both genders, from a pediatric outpatient clinic in southern Brazil. A sociodemographic, behavioral, and food consumption questionnaire was applied, and anthropometric measurements and laboratory blood samples were taken. PON1 arylesterase activity was measured by phenol extinction (U/mL), and DNA extraction and analysis of the PON1 C(‐107)T polymorphism were performed. The Hardy–Weinberg equilibrium was tested with the chi‐squared test and linear regression was used to estimate PON1 activity according to four adjustment models, with an acceptable error of 5%. Results: In the sample, the male gender accounted for 50.5%, 39.2% were 6 years of age, 54.5% had normal weight, and 51.5% had PON1 activity below the median (90.0, 15–30 U/mL). Genotype frequency was 54.6% (53/97), 31.0% (30/97), and 14.4% (14/97), respectively, for CT, CC, and TT, consistent with the Hardy–Weinberg equilibrium (p = 0.22). In the regression analysis, the model that included sociodemographic variables as well as frequency of consumption of fruits, vegetables, legumes, dairy products, and beans estimated a variability of 14.8% in PON1 activity combined with the PON1 C(‐107)T polymorphism. Conclusions: During childhood, a good‐quality diet with greater inclusion of healthy foods was important to predict the activity of the cardioprotective enzyme PON1 combined with the C(‐107)T polymorphism of the PON1 gene. Resumo: Objetivo: A enzima cardioprotetora Paraoxonase 1 (PON1) sofre importante influência de polimorfismos genéticos e fatores nutricionais. O objetivo deste estudo foi investigar a influência da alimentação, do estado nutricional e do polimorfismo C(‐107)T sobre a atividade arilesterase da PON1 em crianças. Métodos: Estudo transversal com 97 crianças entre 5 e 8 anos, de ambos os sexos, de um ambulatório de pediatria no sul do Brasil. Realizou-se questionário sociodemográfico, de comportamento e de consumo alimentar, medidas antropométricas e coleta de sangue em laboratório. A atividade arilesterase da PON1 foi mensurada pela extinção de fenol (U/mL), realizada extração do DNA e análise do polimorfismo PON1 C(‐107)T. O equilíbrio de Hardy‐Weinberg foi testado com qui‐quadrado e usada regressão linear para estimar a atividade da PON1 segundo quatro modelos de ajuste, erro aceitável de 5%. Resultados: Na amostra o sexo masculino representou 50,5%, 39,2% tinham 6 anos, 54,5% eram eutróficos e 51,5% tinha atividade da PON1 inferior à mediana (90,0;15‐30 U/ml). A frequência dos genótipos foi 54,6% (53/97), 31,0% (30/97) e 14,4% (14/97), respectivamente, para CT, CC e TT, estiveram em equilíbrio de Hardy‐Weinberg (p = 0,22). Na análise de regressão o modelo que incluiu variáveis sociodemográficas, de frequência do consumo de frutas, verduras, legumes, laticínios e feijões estimou uma variabilidade de 14,8% na atividade da PON1 combinada ao polimorfismo PON1 C(‐107)T. Conclusões: Na infância uma alimentação de boa qualidade, com maior participação de alimentos saudáveis foi importante para predizer a atividade da enzima cardioprotetora PON1 combinada ao polimorfismo C(‐107)T do gene da PON1.

Overexpression of growth hormone (GH) in gh-transgenic zebrafish of a highly studied lineage F0104 has earlier been reported to cause increased muscle growth. In addition to this, GH affects a broad range of cellular processes in transgenic fish, such as morphology, physiology, and behavior. Reports show changes such as decreased sperm quality and reduced reproductive performance in transgenic males. It is hypothesized that microRNAs are directly involved in the regulation of fertility potential during spermatogenesis. The primary aim of our study was to verify whether gh overexpression disturbs the sperm miRNA profile and influences the sperm quality in transgenic zebrafish. We report a significant increase in body weight of gh-transgenic males along with associated reduced sperm motility and other kinetic parameters in comparison to the non-transgenic group. MicroRNA transcriptome sequencing of gh-transgenic zebrafish sperms revealed expressions of 186 miRNAs, among which six miRNA were up-regulated (miR-146b, miR-200a-5p, miR-146a, miR-726, miR-184, and miR-738) and sixteen were down-regulated (miR-19d-3p, miR-126a-5p, miR-126b-5p, miR-22a-5p, miR-16c-5p, miR-20a-5p, miR-126b-3p, miR-107a-3p, miR-93, miR-2189, miR-202–5p, miR-221–3p, miR-125a, miR-125b-5p, miR-126a-3p, and miR-30c-5p) in comparison to non-transgenic zebrafish. Some of the dysregulated miRNAs were previously reported to be related to abnormalities in sperm quality and reduced reproduction ability in other species. In this study, an average of 134 differentially expressed miRNAs-targeted genes were predicted using the in silico approach. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis demonstrated that the genes of affected pathways were primarily related to spermatogenesis, sperm motility, and cell apoptosis. Our results suggested that excess GH caused a detrimental effect on sperm microRNAome, consequently reducing the sperm quality and reproductive potential of zebrafish males.

The objective of this study was to determine the ability of prostaglandin E2 (PGE2) to induce ovulation and expression of PGE2 receptor (EP2 and EP4) and COX genes (COX-1 and COX-2) in the ovary and pituitary of prepubertal mice. The positive control consisted of the application of 5 μg of gonadotropin-releasing hormone (GnRH, n = 29); the negative control applied 0.5 mL of phosphate buffered saline (PBS, n=31); the treatment tested the application of 250 μg of PGE2 (n = 29), making a total of 89 prepubertal mice (BALB/c). Mice were euthanized 14 to 15 h after treatments to detect ovulation and tissue collection. A Chi-square test was used to compare the proportion of animals ovulating. Gene expressions and number of ovulation were analyzed by one-way ANOVA and Tukey’s test was used to compare means among groups. A greater proportion of mice (P < 0.001) ovulated after receiving GnRH (89.7%, 26/29) compared to PGE2 group (58.6%, 17/29). However, the proportion was higher compared to those treated with PBS (0%, 0/31). Ep2 gene expression in the pituitary was > two-fold higher (P < 0.05) in the PGE2 group compared to the PBS and GnRH groups. Further, PGE2 stimulated Cox1 (2.7 fold, P < 0.05) while GnRH stimulated Cox2 expression (6.5 fold, P < 0.05) in the pituitary when compared to the PBS group. In conclusion, our results support the hypothesis that PGE2 can induce ovulation in prepubertal mice with a concomitant increase in Ep2 and Cox1 gene expression in the pituitary gland.

This study aimed to evaluate the role of prostaglandin F2α (PGF) on ovulation. In Experiment 1, cows were randomly allocated to two treatments to receive 150 μg of d-Cloprostenol (PGF Group, n = 12) or 2 mL of NaCl 0.9% (Control Group, n = 11) and CIDRs, were removed 4 days later. No cow ovulated in Control and PGF groups. In Experiment 2, cows were randomly separated into two experimental groups to receive 4 injections of 150 μg of d-Cloprostenol (n = 9) or 2 mL of NaCL 0.9% (n = 9). In this experiment, ovulation was not observed in any cows. In Experiment 3, ovariectomized cows receive three injections of 300μg of PGF analog (PGF Group, n = 5), 100μg of Lecirelin (GnRH Group, n = 5) or 2 mL of PBS (Control Group, n = 4). The LH concentration was higher (P 11.5 mm) were treated with Saline (Control Group, n = 6); Lecirelin (GnRH Group, n = 7) or Cloprostenol Sodium (PGF Group, n = 6). There was a significant increase in the vascular area of follicles from 0 to 24 h in GnRH and PGF treatments. In conclusion, PGF was not able to induce ovulation in cows with high or low plasma progesterone concentration. Additionally, PGF alone was not able to induce LH release and follicle luteinization, but increased follicular vascularization.

This study evaluated the effect of a prostaglandin F2? (PGF) analogue as an ovulatory stimulus in dairy cows and buffaloes raised in the Amazon biome. To this end, three experiments were performed in the state of Rondônia, located in the Amazon biome. In Experiment 1, 22 lactating dairy buffaloes received 2 mg of intramuscular (I.M.) estradiol benzoate (EB) on day 0 and an intravaginal progesterone-releasing device (CIDR) from day 0 to day 9 of the protocol. On days 8 and 9, all cows were given 500 ?g of I.M. d-cloprostenol (PGF analogue). On day 10, buffaloes were divided into two groups to receive 500 ?g of PGF (PGF group, n = 8) or no treatment (CTL group, n = 14), respectively. In Experiment 2, 16 lactating crossbred dairy cows (Holstein x Gir) received 2 mg of EB on day 0 and a CIDR insert from day 0 to day 8. On days 7 and 8, all cows were given 500 ?g of d-cloprostenol. On day 9, cows were divided into two groups to receive 500 ?g of d-cloprostenol (PGF group, n = 8) or no treatment (CTL group, n = 8), respectively. In Experiment 3, 16 lactating crossbred dairy cows (Holstein x Gir) were handled and treated similarly as in Experiment 2, although cows did not receive d-cloprostenol on day 8. Single-point outcome variables were analyzed using one-way analysis of variance (ANOVA), while proportions with dichotomous outcomes were analyzed with the chi-square test. In Experiment 1, there was no difference (P = 0.30) in the ovulation rate between groups, and, on average, 68% of the buffaloes ovulated. Moreover, the treatment did not affect the interval to ovulation (P = 0.61) nor the diameter of the preovulatory follicle (P = 0.47). As for Experiment 2, only one cow, from the PG group, did not ovulate. There were no differences between the CTL and PG groups (P = 0.69) in the moment of ovulation, which occurred in average 82 h after CIDR removal. Finally, in Experiment 3, cows treated with PGF ovulated earlier than those in the CTL group (62.5 ± 5.8 and 94.5 ± 13.5 h, respectively; P = 0.05). Collectively, these results suggest that PGF hastens ovulation in lactating dairy cows, whereas no effect was observed in dairy buffaloes.

Crohn’s disease (CD) and rheumatoid arthritis (RA) are immune mediated inflammatory diseases. Several studies indicate a role for microRNAs (miRNAs) in the pathogenesis of a variety of autoimmune diseases, including CD and RA. Our study’s goal was to investigate circulating miRNAs in CD and RA patients to identify potential new biomarkers for early detection and personalized therapeutic approaches for autoimmune diseases. For this study, subjects with CD (n = 7), RA (n = 8) and healthy controls (n = 7) were recruited, and plasma was collected for miRNA sequencing. Comparison of the expression patterns of miRNAs between CD and healthy patients identified 99 differentially expressed miRNAs. Out of these miRNAs, 4 were down regulated, while 95 were up regulated. Comparison of miRNAs between RA and healthy patients identified 57 differentially expressed miRNAs. Out of those, 12 were down regulated, while 45 were up regulated. For all the miRNAs down regulated in CD and RA patients, 420 GO terms for biological processes were similarly regulated between both groups. Therefore, the identification of new plasma miRNAs allows the emergence of new biomarkers that can assist in the diagnosis and treatment of CD and RA.

The aim of this study was to compare the J-Synch and conventional protocols associated with estrus detection in beef heifers and to compare pregnancy rate between non-lactating cows displaying estrus or not during the J-Synch protocol. In Experiment 1, heifers were subjected to timed artificial insemination (AI) in a conventional protocol with ECP (n=147) or J-Synch protocol plus eCG (n=149). The AI occurred 12 hours after estrus expression; or 48 (Conventional protocol) and 72 hours (J-Synch protocol) after device removal for animals not displaying estrus. The J-Synch group received 10 µg of GnRH at AI. In Experiment 2, the JSynch was performed (n=116 cows), but without eCG injection, and estrus was monitored. Pregnancy rate was not different between protocols in Experiment 1 (Conventional: 50.68%; J-Synch: 60.4%). Heifers that displayed estrus had higher pregnancy rate only in the conventional protocol. In Experiment 2, pregnancy rate was not different between cows that displayed estrus or not. Therefore, performing AI earlier according to estrus expression increases pregnancy rate in conventional protocol, however it does not increase pregnancy rate in the J-Synch protocol.

Abstract Head and neck cancer is characterized by malignant tumors arising from the epithelium covering the upper aerodigestive tract, and the majority of these epithelial malignancies are squamous cell carcinomas (SCCs) of the oral cavity (OSCCs). The aim of the current work was to identify miRNAs regulated in OSCC cancerous tissue when compared to a healthy adjacent tissue and to verify the presence of the same miRNAs in the circulation of these patients. For that serum samples and biopsies of healthy and tumor tissues were collected from five patients diagnosed with OSCC of the oral cavity, RNA was extracted from these samples and microRNAs libraries were prepared and sequenced. A total 255 miRNAs were identified in tissue and 381 different miRNAs were identified in serum samples. When comparing the miRNA expression between tumor and healthy tissue we identified 48 miRNAs (25 down- and 23 up-regulated) that were differentially expressed (FDR

A ação do hormônio Lactogênio Placentário Bovino (bLP) no metabolismo materno ainda é pouco conhecida. Alguns marcadores, como a proteína de fase aguda Paraoxanase (PON1), são utilizados como indicadores da função hepática auxiliando na determinação da condição metabólica no período de transição em vacas leiteiras. O objetivo deste trabalho foi avaliar a atividade sérica de PON1 durante o periparto em vacas leiteiras com diferentes níveis de bLP. As vacas foram divididas em três grupos de acordo com as concentrações plasmáticas de bLP, em que BAIXO < 2,68 ng mL-1 (n=6), MÉDIO > 2,68 ng mL-1 e < 2,80 ng mL-1 (n=6) e ALTO > 2,80 ng mL-1 (n=6). O período experimental ocorreu entre os 21 dias pré-parto e 28 dias pós-parto. Amostras de soro foram coletadas para a determinação das concentrações de bLP e atividade sérica de PON1. Houve diferença entre os três grupos (P ? 0,0001) de acordo com as concentrações de bLP, assim como entre os dias coletados (P ? 0,0001). No pré-parto, os níveis de PON1 apresentaram diferença entre grupos (P ? 0,05) e coletas (P ? 0,05). Vacas com alta concentração de bLP apresentaram menor atividade sérica de PON1 (P ? 0,05), enquanto vacas com baixos níveis do hormônio obtiveram maior atividade da enzima (P ? 0,05). No pós-parto não houve diferença entre grupos (P ? 0,10), houve diferença entre os dias coletados (P ? 0,0001) e interação entre grupos e coletas (P ? 0,01). O grupo com altas concentrações de bLP apresentou menores níveis de PON1 (P ? 0,01), enquanto vacas do grupo com baixo bLP mantiveram maiores concentrações de PON1 (P ? 0,01). Conclui-se que, vacas com maiores concentrações de bLP no período pré-parto apresentam redução na atividade sérica da enzima PON1 durante o período periparto.

Head and Neck Squamous Cell Carcinoma (HNSCC) is the sixth most common cancer worldwide. These tumors originate from epithelial cells of the upper aerodigestive tract. HNSCC tumors in different regions can have significantly different molecular characteristics. While many microRNAs (miRNAs) have been found to be involved in the regulation of the carcinogenesis and pathogenesis of HNSCC, new HNSCC related miRNAs are still being discovered. The aim of this study was to explore potential miRNA biomarkers that can be used to diagnose HNSCC and prognose survival of HNSCC patients. For this purpose, we chose a panel of 12 miRNAs: miR-146a-5p, miR-449a, miR-126-5p, miR-34a-5p, miR-34b-5p, miR-34c-5p, miR-217-5p, miR-378c, miR-6510-3p, miR-96-5p, miR-149-5p, and miR-133a-5p. Expression of these miRNAs was measured in tumor tissue and neighboring healthy tissue collected from patients diagnosed with HNSCC (n = 79) in either the oral cavity, oropharynx, or larynx. We observed a pattern of differentially expressed miRNAs at each of these cancer locations. Our study showed that some of these miRNAs, separately or in combination, could serve as biomarkers distinguishing between healthy and tumor tissue, and their expression correlated with patients’ overall survival.

This study evaluated the relationship of pre-partum BCS variations on the acute phase response around parturition and milk production of dairy cows. The animals were divided into two groups: cows that gained BCS from the third to the first week pre-partum (GBC = n 11) and those that lost BCS (LBC = n 9) in the same period. Blood samples were collected pre and post-partum to determine serum concentrations of glucose (GLU), non-esterified fatty acids (NEFA), insulin (INS), albumin (ALB), paraoxonase (PON), haptoglobin (HP) and blood cell counts. The GBC group showed higher PON concentration in the postpartum (p

Oral squamous cell carcinoma (OSCC) is the most common type of head and neck cancer and, as indicated by The Oral Cancer Foundation, kills at an alarming rate of roughly one person per hour. With this study, we aimed at better understanding disease mechanisms and identifying minimally invasive disease biomarkers by profiling novel small non-coding RNAs (specifically, tRNA halves and YRNA fragments) in both serum and tumor tissue from humans. Small RNA-Sequencing identified multiple 5′ tRNA halves and 5′ YRNA fragments that displayed significant differential expression levels in circulation and/or tumor tissue, as compared to control counterparts. In addition, by implementing a modification of weighted gene coexpression network analysis, we identified an upregulated genetic module comprised of 5′ tRNA halves and miRNAs (miRNAs were described in previous study using the same samples) with significant association with the cancer trait. By consequently implementing miRNA-overtargeting network analysis, the biological function of the module (and by “guilt by association,” the function of the 5′ tRNA-Val-CAC-2-1 half) was found to involve the transcriptional targeting of specific genes involved in the negative regulation of the G1/S transition of the mitotic cell cycle. These findings suggest that 5′ tRNA-Val-CAC-2-1 half (reduced in serum of OSCC patients and elevated in the tumor tissue) could potentially serve as an OSCC circulating biomarker and/or target for novel anticancer therapies. To our knowledge, this is the first time that the specific molecular function of a 5′-tRNA half is specifically pinpointed in OSCC.

The transition period of dairy cows is characterized by several metabolic alterations, due to the negative energy balance, which negatively affects fertility. The aim of this study was to evaluate the effect of supplementing the in vitro maturation medium with serum from dairy cows at early and late lactation in the oocyte maturation on the rates of nuclear maturation, cleavage and initial embryo development. The cumulus-oocyte complexes (COCs) were randomly divided into treatments: LL: serum from four cows at 354 ± 3.8 days in milk (DIM); and EL: serum from four cows at 10.8 ± 4.8 DIM. Subsequently, the COCs were submitted to maturation, fertilization; and in vitro culture in an incubator at 39°C with 5% CO2. For analysis of nuclear maturation; oocytes were collected after IVM, denuded and stained with Hoechst®. No difference was observed in the number of oocytes that reached metaphase II among treatments. The cleavage rate was not different between LL (59.3%; 320/540) and EL (47.0%; 243/517) treatments (P>0.05), as embryonic development between LL (15.0%; 81/540) and EL (16.6%; 86/517) (P>0.05). It was concluded that supplementation with serum from cows at different lactation periods during the bovine oocyte IVM did not influence the results of in vitro embryo production.

ABSTRACT Objective The aims of this study were to investigate changes in serum paraoxonase 1 (PON1) activity in women at the pre and postmenopausal stages and its association with the PON1 C(-107)T polymorphism and food intake profile. Subjects and methods A cross-sectional study with female patients aged between 35 and 59 years old was conducted. Women were divided into two groups: premenopausal (n = 40) and postmenopausal (n = 36). Women enrolled in the study had serum PON1, total cholesterol, HDL, LDL, glucose and HbA1c, as well as the BMI measured. Additionally, women were genotyped for the PON1 T(-107)C polymorphism and the food intake profile was obtained through interview. Results Glucose (p = 0.03), HbA1c (p = 0.002) and total cholesterol (p = 0.002)concentrations were higher in post than premenopausal women, however PON1 activity was not different (p > 0.05). Carriers of the C allele had higher PON1 activity (CC: 88.9 ± 6.5 U/mL and CT: 79.9 ± 4.7 U/mL) than women of the TT genotype (66.6 ± 5.9 U/mL) (p < 0.05). However, the model predicting PON1 activity was slightly better when genotype, total fat and cholesterol content in the diet were all included. Conclusion In sum, we observed that the PON1 C(-107)T genotype was the major regulator of PON1 activity, and menopause had no effect on PON1 activity. The lipid and glycemic profile were altered in postmenopausal women.

Colorectal cancer is recognized as the fourth leading cause of cancer-related deaths worldwide. Thus, there is ongoing search for potential new biomarkers allowing quicker and less invasive detection of the disease and prediction of the treatment outcome. Therefore, the aim of our study was to identify colorectal cancer specific miRNAs expressed in cancerous and healthy tissue from the same patient and to further correlate the presence of the same miRNAs in the circulation as potential biomarkers for diagnosis. In the current study we detected a set of 40 miRNAs differentially regulated in tumor tissue when comparing with healthy tissue. Additionally, we found 8 miRNAs differentially regulated in serum of colorectal cancer patients. Interestingly, there was no overlap in miRNAs regulated in tissue and serum, suggesting that serum regulated miRNAs may be not actively secreted from colorectal tumor cells. However, four of differentially expressed miRNAs, including miR-21, miR-17, miR-20a and miR-32 represent the miRNAs characteristic for different tumor types, including breast, colon, lung, pancreas, prostate and stomach cancer. This finding suggests important groups of miRNAs which can be further validated as markers for diagnosis of tumor tissue and regulation of carcinogenesis.

ABSTRACT: Previous studies have evaluated the effects of different reproductive procedures on discomfort markers in sheep and cattle. Such studies may help stimulate the adoption of techniques that are more beneficial for animal welfare. However, markers that are commonly used to evaluate discomfort are highly influenced by external factors. To overcome this, several systemic markers can be evaluated to more precisely identify stress, pain, and inflammation. Such markers include cortisol, acute phase proteins, bradykinin, and substance P. We aimed to review the potential markers of stress, pain, and inflammation, and discuss how and when they are regulated after different stimuli related to reproductive procedures in cattle and sheep. Furthermore, we aimed to review how reproductive procedures with different degrees of invasiveness cause stress and provide information that may help develop strategies to limit animal discomfort.

ABSTRACT This study aimed to evaluate the effect of Ilex paraguariensis infusion on redox state of Wistar rats submitted to high-fat and standard diet. Glutathione determination and lipid peroxidation in the hippocampus tissues and liver was performed, as well as the analysis of gene expression of superoxide dismutase, catalase and glutathione peroxidase by real-time polymerase chain reaction. The results from hippocampus showed that the groups fed with standard diet exhibited significant reduction of lipid peroxidation when supplemented with Ilex paraguariensis. The analysis from glutathione determination in the hippocampus showed a significant increase in glutathione activity in the group treated with high-fat diet and Ilex paraguariensis. In the liver, results showed no significant difference in both glutatione and lipid peroxidation analisys. Gene expression of superoxide dismutase, catalase and glutathione peroxidase showed that there was significant difference in the groups treated with high-fat diet and Ilex paraguariensis. In summary, the Ilex paraguariensis showed substantial potential for antioxidant activities.

ABSTRACT: The aim of this study was to estimate neosporosis seroprevalence and its associated risk factors in milk herds (Bos taurus taurus) located in the northwestern region of Rio Grande do Sul State, Brazil. Three hundred twenty-two blood samples were collected from dairy cows on 18 farms in 17 cities of this region. An epidemiologic questionnaire was completed for each farm. It consisted of questions about the general characteristics of the herd, reproduction, and animal management. Serum samples were tested for Neospora caninum using a commercial enzyme-linked immunosorbent assay (ELISA) kit. Results indicated a seroprevalence of Neospora in 88.9% (16/18) of herds and 31.1% (100/322) of individuals. Risk factor analyses demonstrated that culling by reproductive disorder (OR = 0.6), flooding (OR = 0.5), and commercial sale (OR = 0.4) were associated with seroprevalence. Nevertheless, the purchase of replacement animals in the herd played an important role in disease occurrence (OR = 2.2). Results of this study suggested that Neospora caninum was present in the studied herds under investigation and that there are risk factors associated with its seroprevalence on the farms of the northwestern of Rio Grande do Sul.

The head and neck squamous cell carcinoma (HNSCC) represents one of the most common cancers in humans. Close to 600,000 new diagnoses are made every year worldwide and over half of diagnosed patients will not survive. In view of this low survival rate, the development of novel cell-based assays for HNSCC will allow more mechanistic approaches for specific diagnostics for each individual patient. The cell-based assays will provide more informative data predicting cellular processes in treated patient, which in effect would improve patient follow up. More importantly, it will increase the specificity and effectiveness of therapeutic approaches. In this study, we investigated the role of serum from HNSCC patients on the regulation of microRNA (miRNA) expression in exposed cells in vitro. Next-generation sequencing of miRNA revealed that serum from HNSCC patients induced a different miRNA expression profile than the serum from healthy individuals. Out of 377 miRNA detected, we found that 16 miRNAs were differentially expressed when comparing cells exposed to serum from HNSCC or healthy individuals. The analysis of gene ontologies and pathway analysis revealed that these miRNA target genes were involved in biological cancer-related processes, including cell cycle and apoptosis. The real-time PCR analysis revealed that serum from HNSCC patients downregulate the expression level of five genes involved in carcinogenesis and two of these genes—P53 and SLC2A1—are direct targets of detected miRNAs. These novel findings provide new insight into how cancer-associated factors in circulation regulate the expression of genes and regulatory elements in distal cells in favor of tumorigenesis. This has the potential for new therapeutic approaches and more specific diagnostics with tumor-specific cell lines or single-cell in vitro assays for personalized treatment and early detection of primary tumors or metastasis.

The objective of this study was to determine the effects of uterine and systemic inflammatory responses to uterine bacterial contamination at calving in dairy cows on the growth and ovulatory outcomes of the first dominant follicle postpartum. Ovulatory capability of the first dominant follicle postpartum was predicted in 53 multiparous cows by using a combination of follicle growth characteristics and circulating estradiol concentrations. Endotoxin levels were assayed in follicular fluid samples that were aspirated the day after ovulatory outcome prediction. Plasma levels of haptoglobin, a proinflammatory acute phase protein, and paraoxonase, a negative acute phase protein were determined. Uterine bacteria and inflammation were evaluated in three uterine fluid samples from each cow collected on the day of calving, the day after follicle aspiration, and at 35 days postpartum. Cows that had a strong initial uterine inflammatory response (robust recruitment of polymorphonuclear leukocytes of ≥ 35% and cows with uterine pH < 8.5 on the day of calving) were more likely to have an ovulatory first dominant follicle. Follicular fluid endotoxin levels were higher in non-ovulatory cows compared with ovulatory cows. Endotoxin levels in circulation were not different between ovulatory groups but were higher prepartum than on day 7 and 14 postpartum. Systemic inflammation characterized by elevated haptoglobin concentrations was higher in non-ovulatory cows despite similar bacterial contamination and circulating endotoxin levels. Paraoxonase activity in follicular fluid was significantly associated with the paraoxonase activity in plasma, however, plasma paraoxonase concentrations were not different between non-ovulatory and ovulatory cows. Cows with a higher uterine bacterial load on the day of calving had slower ovarian follicle growth. In summary, a robust uterine inflammatory response on the day of calving was positively associated with ovarian function while elevated systemic inflammation during the early postpartum period was negatively associated with the ovulatory status of the first dominant follicle postpartum.

The Ames dwarf (df/df) mice have extended longevity and can preserve the ovarian reserve longer than Normal (N) mice. Based on this, the aim of our study was to evaluate the ovarian microRNA (miRNA) profile in young and aged df/df and N mice. Ovarian tissue was collected at 5-6 months and at 21-22 months of age for miRNA sequencing. We detected a total of 404 miRNAs in the ovarian samples, from which the abundance of 22 and 33 miRNAs changed with age in N and df/df mice, respectively. Of these, only three miRNAs were commonly regulated with age between N and df/df mice, indicating a very divergent miRNA profile between genotypes. We also detected that 46 miRNAs were regulated between N and df/df mice, of which 23 were regulated exclusively in young mice, 12 exclusively in old mice and 12 commonly regulated at young and old ages. Many genes likely to be targeted by these miRNAs are involved in the FoxO, mTOR, PI3k/Akt and insulin signaling pathways. These results suggest that the aging process has a differential impact on the ovarian miRNA profile in df/df mice, and suggest that these miRNAs can be central players in the maintenance of a younger ovarian phenotype.

The liver is an organ with a differential value on the market. However, due to its metabolic functions it is susceptible to various types of alterations, including a large rate of disposal by perihepatitis. The aim of this study was to evaluate the sensitivity and specificity of the diagnosis of perihepatitis by the Federal Inspection Service (FIS), according to histopathological examination and correlate these findings with plasmatic concentrations of AST and GGT. One hundred and fifty four culled sows of the Landrace breed were used. Slaughter was performed by the method of desensitization by electrical stimulation and subsequent exsanguination. Then 5 mL of whole blood was collected to evaluate concentrations of GGT and AST. During evisceration, liver condition was assessed by visual inspection and classified as good or condemned by perihepatitis. Also, fragments of the liver were collected to histopathologic examination. The alterations in the liver parenchyma were classified as degenerative, inflammatory or reparative. The alterations found in liver tissues, considered as perihepatitis by visual inspection, were often due to fibrosis, fatty degeneration or mild hepatitis. The serum concentrations of GGT were high. Moreover, it concentrations were higher in livers with lesions of inflammatory and degenerative nature when compared to livers without injuries (p0.05). Concluding, the perihepatitis diagnosis from the FIS showed low specificity and sensitivity, and the enzyme GGT was a good indicator of liver injuries in swine.

The objective of this study was to evaluate the influence of the ovulatory follicle diameter on the reproductive performance of lactating beef cows subjected to low progesterone fixed-time artificial insemination (FTAI) protocols. Ninety-three lactating beef cows (60-80 days postpartum) at random stages of estrous cycle were given a luteolytic dose of prostaglandin F2α (500 μg cloprostenol; PGF) twice, 11 d apart. Ten days after the second PGF treatment, cows were given 1.5 mg of estradiol benzoate im and a progesterone-releasing intravaginal device (Cue-Mate) with a single pod containing 0.78 g progesterone (Day 0). Cows received another luteolytic dose of PGF on Day 0. On Day 8, the Cue-Mate was removed. Fifty-four to 56 hours later, cows received 12.5 mg of porcine LH (pLH) i.m. and were concurrently artificially inseminated. Ultrasound examinations of the ovaries were performed on Days 10 and 17 to evaluate the diameter of ovulatory follicle and corpus luteum, respectively. Cows which presented ovulatory follicle > 19 mm resulted in larger corpus luteum than cows that had ovulatory follicle

The objective of this study was to evaluate the plasma concentrations of insulin-like growth factor-I (IGF-I), and the mRNA hepatic expression of IGF-I and of the growth hormone receptors GHR and GHR 1A, in postpartum beef cows. Four Angus and four crossbred (Angus x Nelore) postpartum suckled beef cows were used. Liver and blood samples were collected every 10 days, from calving to 40 days postpartum, for gene expression and for β-hydroxybutyrate and IGF-I assays, respectively. Samples for progesterone assay were collected every other day, from day 10 to 40 postpartum. Three cows ovulated before 40 days postpartum. IGF-I concentration was higher in Angus x Nelore than in Angus cows. There was no difference in the expression of GHR, GHR 1A and IGF-I according to breed or ovulatory status. IGF-I concentrations were higher in crossbred cows, but have not changed according to postpartum ovulatory status. Moreover, changes in postpartum IGF-I concentrations are not associated with changes in liver GHR, GHR 1A and IGF-I mRNA expression in either breed.O objetivo deste estudo foi avaliar as concentrações plasmáticas do fator de crescimento semelhante à insulina tipo I (IGF-I) e a expressão hepática de mRNA de IGF-I e dos receptores do hormônio de crescimento GHR e GHR 1A, no pós-parto de vacas de corte. Quatro vacas Angus e quatro mestiças (Angus x Nelore) lactantes foram usadas durante o período de pós-parto. As coletas de tecido hepático e de sangue foram realizadas a cada dez dias, do parto até 40 dias pós-parto, para a avaliação da expressão gênica, e análise de β-hidroxibutirato e IGF-I, respectivamente. As amostras para análise de progesterona foram coletadas a cada dois dias, dos dez aos 40 dias pós-parto. Três vacas ovularam antes dos 40 dias pós-parto. A concentração de IGF-I foi maior em vacas Angus x Nelore do que em vacas Angus. Não houve diferença na expressão hepática de GHR, GHR 1A e IGF-I de acordo com a raça ou com a ovulação. As concentrações de IGF-I foram maiores em vacas cruzadas, e não mudaram com o status ovulatório pós-parto. Além disso, as mudanças observadas na concentração de IGF-I, durante o período pós-parto, não estão associadas a alterações na expressão hepática de mRNA de GHR, GHR 1A e IGF-I, em nenhuma das duas raças.

Foi realizado um estudo para identificar parâmetros de desempenho associados a indicadores de eficiência durante o parto de fêmeas suínas. Foram coletados os seguintes dados de 636 partos e 7.100 leitões nascidos: ordem de parto, número de leitões nascidos vivos, natimortos, mumificados, total de nascidos, aplicação de ocitocina, uso de palpação vaginal, momento do nascimento de cada leitão e do início e final do parto. O intervalo médio de nascimento dos leitões foi de 16,7 minutos e a duração média do parto, de 247 minutos. A duração média do parto aumentou conforme o total de leitões nascidos, o uso de ocitocina e a ocorrência de palpação vaginal. O total de nascimentos nos partos longos (> 211 min), de 13,1 leitões, foi maior que nos partos curtos (< 210 min), de 11,7 leitões, portanto, quanto maior o total de nascidos, maior a duração do parto. As leitegadas foram maiores nas fêmeas de segundo parto (12,8) em comparação àquelas de primeiro parto (11,9). A duração do parto é influenciada pelo total de leitões nascidos, que é maior em fêmeas de segundo parto e está associada à maior ocorrência de leitões natimortos e mumificados.This study aimed to identify some performance parameters associated with the farrowing efficiency in sows. From a total of 636 births and 7,100 piglets born, data was collected for: parity (OP), number of piglets born alive (NV), stillborns (NAT), mummified (MUM), total litter size (TN), use of vaginal palpation and oxytocin injection, time of birth for each piglet, and beginning and end of the parturition. The average interval among piglets birth was 16.7 min and the average duration of parturition was 247 min. The average duration of parturition increased regarded to the number of born piglets, the use of oxytocin and with the ocurence of vaginal palpation. The total number of births in the long parturitions (> 211 min), with 13.1 piglets was higher than in the short parturitions (< 210 min), with 11.7 piglets, therefore, the higher the number of total piglets born, the longer is the duration of parturition. Litter size was higher for sows of the second parity (12.8) than for sows of first parity (11.9). The duration of parturition is affected by the number of piglets born, which is higher in second parity than first parity females and it is associated to the higher occurence of stillbirths and mummified piglets.

This study aim to evaluate the quality of boar semen stored at different temperatures and media on PIGPEL5 extender, using egg yolk as an external membrane protector, and on PIGPEL5Plus, containing low density lipoprotein (LDL) as protector. In the experiment 1, the semen doses were stored in a semen container at 5 and 17 °C and the results were compared with semen stored at 17 °C in BTS extender. In the experiment 2, the doses with the PIGPEL5 and PIGPEL5Plus extenders were stored in a commercial refrigerator and in semen container, both at 5 °C. The use of extenders with extracellular cryoprotectant (PIGPEL5 e PIGPEL5Plus) showed higher (p < 0.05) motility values in comparison to BTS until 48 h. Furthermore, the use of extracellular cryoprotectant was good to maintenance of membrane integrity and normal morphology at 5 oC similarly to 17 oC extenders. There was no difference (p > 0.05) in the motility, membrane integrity and morphology for the substitution of egg yolk (PIGPEL5) for LDL (PIGPEL5Plus). In experiment 2, also there was no difference (p > 0.05) in motility, membrane integrity and morphology between extenders and type of container until 72 h. The conservation of semen on PIGPEL5 extender and commercial refrigerator (5 ºC) allowed patterns of sperm quality according recommended for AI.Este estudo avaliou a qualidade do sêmen suíno preservado em diferentes temperaturas e meios de conservação no diluente PIGPEL5, utilizando gema de ovo como protetor externo de membrana, e no diluente PIGPEL5Plus, utilizando lipoproteína de baixa densidade (LDL) como protetor. No experimento 1, as doses de sêmen foram conservadas em refrigerador de sêmen a 5 e 17 °C nos diluentes PIGPEL5 e PIGPEL5Plus e comparadas ao diluente BTS a 17 °C. Já no experimento 2, foi comparada a conservação do sêmen nos diluentes PIGPEL5 e PIGPEL5Plus em geladeira convencional ou em refrigerador de sêmen a 5 ºC. Observou-se que o uso de diluidores com crioprotetor extracelular (PIGPEL5 e PIGPEL5Plus) apresentou maiores (p < 0,05) resultados de motilidade com relação ao BTS até 48 h. Além do mais, o uso de crioprotetor extracelular foi benéfico para a manutenção da integridade e morfologia normais a temperatura de 5 ºC em níveis similares aos diluidores a 17 ºC. Não houve diferença (p > 0,05) na motilidade, integridade de membrana e morfologia para a substituição da gema de ovo (PIGPEL5) pela LDL (PIGPEL5Plus). No experimento 2, também não houve diferença na motilidade, integridade de membrana e morfologia espermática entre os diluentes e formas de acondicionamento testados até 72 h (p > 0,05). O acondicionamento do sêmen no diluente PIGPEL5 em geladeira doméstica (5 ºC) proporcionou padrões de qualidade espermática de acordo com o recomendado para a IA.

Os objetivos neste experimento foram avaliar o efeito do tempo de exposição a um progestágeno (P4) na taxa de prenhez de novilhas púberes e pré-púberes inseminadas em tempo-fixo (IATF) e estudar o retorno econômico dos protocolos utilizados, por meio da comparação desses protocolos ao sistema tradicional de acasalamento natural aos 26 meses de idade. Foram utilizadas 114 novilhas da raça Bradford com 16 meses de idade, classificadas em quatro grupos de acordo com o tratamento e a maturidade sexual: GPPLP - pré-puberes, longo tratamento com P4 (n=21); e GPPCP - pré-puberes, curto tratamento com (n=19); GPLP - púberes, longo tratamento com P4 (n=34); e GPCP - púberes, curto tratamento com P4 (n=40). As novilhas do GPPLP e GPLP receberam no dia -8 um pessário intravaginal impregnado com acetato de medroxiprogesterona. No dia 0, o pessário foi removido e outro foi inserido, juntamente com uma injeção de 2,5 mg de benzoato de estradiol (BE). No dia 8, o pessário foi retirado e as novilhas receberam 0,5 mg de BE e 250 µg de cloprostenol sódico. A inseminação artificial em tempo fixo (IATF) foi realizada 54 horas depois. As novilhas do GPPCP e GPCP receberam o mesmo tratamento de sincronização dos grupos GPPLP e GPLP, com exceção do AMP do dia -8 ao dia 0. A taxa de prenhez foi maior nas novilhas púberes (60,3%) que nas pré-puberes (35,0%). No entanto, não foi detectado efeito do tempo de tratamento na taxa de prenhez nas novilhas púberes e pré-puberes. A resposta econômica na utilização de protocolos com P4 foi maior em novilhas aos 16 meses que no sistema de acasalamento aos 26 meses. A exposição prolongada a um P4 não foi eficiente para aumentar a taxa de prenhez de novilhas submetidas a IATF.The aim of this experiment was to evaluate the effect of the duration of progestagen (P4) exposure and sexual maturity of prepubertal and pubertal heifers submitted to timed artificial insemination (TAI). A hundred and fourteen 16- month old Bradford heifers were classified, according treatment and sexual maturity, in 4 groups: PPLPG (prepubertal long exposure to P4, n=21), PPSPG (prepubertal short exposure to P4, n=19), PLPG (pubertal long exposure to P4, n=34) and PSPG (pubertal short exposure to P4, n=40). On day -8, the heifers from PPLPG (prepubertal long exposure to progestagen) and PLPG (pubertal long exposure to progestagen) received an intravaginal sponge impregnated with medroxiprogesterone acetate (MAP). On day 0 the sponge was removed and a new sponge was inserted, together with an injection of 2.5 mg estradiol benzoate (EB). On day 8 the sponge was removed and the heifers received an injection of 0.5 mg EB and 250 ìg sodium cloprostenol. The TAI was performed 54 hours after sponge withdrawal. The heifers from PPSPG and PSPG received the same synchronization treatment, except for the additional MAP from day -8 to day 0. The pregnancy rate was higher in pubertal (60.3%) than in pre-pubertal (35.0%) heifers. However, there was no effect of the duration of treatment in the pregnancy rate between pubertal and prepubertal heifers. The second part of this study aimed to evaluate the economical return of the protocols and compare them with the traditional breeding system at 26 months of age. A better economic response with the use of protocols associating P4 in heifers at 16 months was observed compared to the traditional system at 26 months of age. The prolonged exposure to P4 did not improved pregnancy rate in heifers submitted to TAI.

Objetivou-se verificar a redução de perdas embrionárias por meio do bloqueio da secreção de prostaglandina com a utilização de flunixin meglumine (FM) e avaliar o retorno econômico desta técnica. No experimento 1, utilizaram-se vacas de corte no pós-parto e, no experimento 2, somente novilhas. Todas as fêmeas foram sincronizadas e inseminadas em tempo fixo (IATF) e, após 14 dias da IATF, foram distribuídas em dois grupos: grupo flunixin meglumine (GFM), composto de 59 vacas no experimento 1 e 23 novilhas no experimento 2; e grupo controle (GC), de 56 vacas no experimento 2 e 20 novilhas no experimento 1, que receberam ou não flunixin no dia 14, respectivamente. O tratamento com flunixin não influenciou a taxa de prenhez, que, no experimento 1, foi de 42,4% no grupo flunixin e 42,8% no grupo controle e, no experimento 2, de 39,1% no grupo flunixin e 25,0% no grupo controle. Verificou-se rentabilidade de 21,62; 9,24; 14,36 e 12,06% para os protocolos dos grupos controle e flunixin dos experimentos 1 e 2, respectivamente. A taxa de prenhez não foi influenciada pelo uso de flunixin meglumine no dia 14 após inseminação artificial em vacas no pós-parto e novilhas inseminadas em tempo fixo.This study aimed to evaluate the effectiveness and the economical advantage of using flunixin meglumine (FM) to block uterine secretion of prostaglandin in order to reduce embryonic losses in beef cattle females. Postpartum cows (E1) and heifers (E2) were submitted to estrus synchronization and inseminated at fixed time. Fourteen days after the insemination the animals were assigned to the FM treatment group (FMG, including 59 cows and 23 heifers) and to the control group (CG, including 56 cows and 20 heifers). Pregnancy rates (42.4% for FMG and 42.8% for CG in E1) and (39.1% for FMG and 25.0% for CG in E2) did not differ between groups. Estimates of net income were 21.62, 9.24, 14.36 and 12.06% for the CG e FMG protocols in cows and in heifers, respectively. The use of FM at 14 days after AI did not affect pregnancy rates of postpartum beef cows and heifers.

In vitro embryo production (IVP) represents a way to increase gamete use from animals with high zootechnical value. In spite of the advances obtained in IVP over the last few years, production of transferable embryos is still low. The aim of this review is to discuss ways to produce in vitro embryos, as well as oocytes formation and maturation processes that can be related to the effectiveness of obtained results. Some studies show the influence of follicular growth factors, gonadotropins, steroids and other hormones on the follicular development and the quality of the cumulus oocyte complex (COC). The follicular phase of slow growth is critical for the development of the oocyte capacity to reach the final competence and diameter. Information about endocrine influences, or likewise, the dependence of growth of small antral follicles when a loss in the oocyte or follicle functionality occurs is scarce in the literature. A variable number of different techniques and protocols for treatment of oocytes donors are described with the aim of improve the results, the COCs recovering rate and the developmental competence in vitro of collected oocytes. From the considerations presented in this review, it is possible to verify the importance of better understanding the factors involved in the IVP process, with the aim of allow new alternatives to increase the results obtained in programs of animal assisted reproduction.La producción in vitro de embriones (PIV) representa una manera de aumentar el uso de gametos de animales con alto valor zootécnico. A pesar de los avances obtenidos en PIV en los últimos años, la producción de embriones tranferibles sigue siendo baja. El objetivo de esta revisión es discutir maneras de producir embriones in vitro, así como los procesos de formación y de maduración de los oocitos que se pueden relacionar con la eficacia de los resultados obtenidos. Algunos estudios demuestran la influencia de los factores foliculares del crecimiento, gonadotrofinas, esteroides y otras hormonas en el desarrollo folicular y la calidad del complejo del cumulus oocito (CCO). La fase folicular del crecimiento lento es crítica para el desarrollo de la capacidad del oocito de alcanzar la capacidad y el diámetro final. Información sobre influencias endocrinas, o además, la dependencia del crecimiento de pequeños folículos antrales cuando ocurre una pérdida en la funcionalidad del oocito o del folículo, es escasa en la literatura. Un número variable de diversas técnicas y los protocolos para el tratamiento de oocitos de las donantes son descritos en esta revisión, con lo objetivo de mejorar los resultados, el índice de la recuperacion de CCOs y la capacidad de desarrollo in vitro de oocitos recogidos. De las consideraciones presentadas en esta revisión, es posible verificar la importancia de entender los factores implicados en el proceso de PIV, para permitir el desarrollo de nuevas alternativas que mejoren los resultados obtenidos en programas de la reproducción animal asistida.A produção in vitro (PIV) de embriões representa uma maneira de incrementar o uso de gametas de animais de alto valor zootécnico. Apesar dos avanços obtidos na PIV nos últimos anos, a produção de embriões transferíveis ainda é baixa. O objetivo desta revisão é discutir maneiras de produzir embriões in vitro, assim como o processo de formação e maturação de oócitos, que pode estar relacionado a eficácia dos resultados obtidos. Aguns estudos demonstram a influência de fatores de crescimento, gonadotrofinas, esteróides e outros hormônios no desenvolvimento folicular e na qualidade do complexo cumulus oócito (CCO). A fase folicular de crescimento lento é critica para o desenvolvimento da capacidade do oócito de atingir a competência e o diâmetro finais. Informação sobre as influências endócrinas, ou seja, da dependência do crescimento de pequenos folículos antrais quando ocorre perda da funcionalidade do oócito ou folículo são escassas na literatura. Um número variável de diferentes técnicas e protocolos para o tratamento de doadoras de ovóctios são descritos com o objetivo de melhorar os resultados, a taxa de recuperação de CCOs e o desenvolvimento da competência in vitro dos oócitos coletados. Das considerações apresentadas nesta revisão é possível verificar a importância do conhecimento dos fatores envolvidos no processo de PIV, com o objetivo de possibilitar que novas alternativas incrementem os resultados obtidos em programas de reprodução animal assistida.

The aim of this study was to evaluate the use of a prostaglandin F2α analogue (PGF2α) and oestradiol benzo-ate (ODB) in estrus synchronization programs. A hundred and eighty five non-lactating females divided in 4 groups were used. G1 (44) and G2 (46) received two applications of PGF2α (D0 and D11) and G3 (26) and G4 (37) received one application after 5 days of estrous detection. G1 and G3 also received a 1 mg ODB application 48 hours after the PGF2α. The pregnancy rate at AI was 42,2% for G1, 47,8% for G2, 32,1% for G3 e 57,6% for G4 (p>0,05) and of 38,4% and 51,4% for animals submitted to FTAI and estrous detection, respectively (p>0,05). This results show that pregnancy rates of females fixed time inseminated didn’t differ of that inseminated after estrus detection. Key-words: Beef cattle, estrus synchronization, fixed time artificial insemination, prostaglandin. O objetivo deste estudo foi avaliar o emprego de um análogo da prostaglandina F2α (PGF2α) em associação ao benzoato de estradiol (BE) em programas de sincronização de cios. Utilizaram-se 185 fêmeas não-lactantes divididas em quatro grupos. O G1 (44) e o G2 (46) receberam duas aplicações de PGF2α (D0 e D11) e os animais do G3 (26) e G4 (37) uma aplicação de PGF2α, após cinco dias de detecção de cio. O G1 e o G3 receberam 1mg de BE 48 horas após a PGF2α. A taxa de prenhez na IA foi de 42,2% no G1, 47,8% no G2, 32,1% no G3 e 57,6% no G4 (p>0,05) e de 38,4% e 51,4% para animais submetidos à IATF e à detecção de cio, respectivamente (p>0,05). Notou-se que a taxa de prenhez nas fêmeas inseminadas em tempo-fixo não diferiu daquelas que foram inseminadas após a detecção de cio. Palavras-chaves: Bovinos de corte, inseminação artificial em tempo fixo, prostaglandina, sincronização de cios.

O objetivo deste experimento foi avaliar o efeito das técnicas transcervical e laparoscópica sobre a taxa de prenhez de ovelhas inseminadas em tempo-fixo. A taxa de prenhez para as técnicas transcervical e laparoscópica foi de 40% (p>0.05). Quanto ao local de deposição do sêmen na inseminação artificial (IA) transcervical, não houve diferença na taxa de prenhez (p>0.05), com taxas de 25.0% na cervical superficial, 43.7% cervical médio, 41.7% cervical profundo e 50.0% intra-uterino. Estes resultados demonstram que a IA transcervical apresenta resultados semelhantes aos obtidos com o uso de laparoscopia, quando estas técnicas são realizadas em tempo-fixo. PALAVRAS-CHAVE: Cérvix, intrauterino, sêmen congelado, tracionamento The objective of this experiment was to evaluate the effect of the transcervical and laparoscopic techniques on the pregnancy rate of inseminated sheep in fixed-time. The pregnancy rate for the transcervical and for laparoscopic techniques was 40% (p>0.05). About the place of deposition of the semen in transcervical artificial insemination (AI), there was not difference in the pregnancy rate (p>0.05), with rates of 25.0% in the superficial cervical, 43.7% in medium cervical, 41.7% in deep cervical and 50.0% intrauterine. These results demonstrate that transcervical AI showed similar results to the obtained with laparoscopy, when these techniques are accomplished in fixed-time. KEY WORDS: Cervix, intrauterine, frozen semen, traction

The aim of this work was to validate a protocol for induction of subacute ruminal acidosis (SARA) (Experiment 1) and test the efficiency of probiotic Saccharomyces cerevisiae or monensin to avoid pH ruminal drops in sheep (Experiment 2). In Experiment 1, six ewes were fasted for two days and then fed most with concentrate during four days. Ewes in this protocol had ruminal fluid pH below 6.0 and kept it for 75 consecutive hours. In Experiment 2, 18 sheep were distributed into three groups: Control (CG, n = 6), monensin (MG, n = 6) and probiotic group (PG, n = 6). SARA was induced according Experiment 1. PG had lower pH (5.7 ± 0.1) than CG (6.0 ± 0.1) (P = 0.05), while MG (5.7 ± 0.1) was similar to both during SARA induction. SARA induction reduced ruminal protozoa population (P < 0.05) and increased chloride concentrations in ruminal fluid (P < 0.01). In serum, SARA increased concentrations of phosphorus (P < 0.01), AST (P < 0.01) and GGT (P < 0.01), but reduced LDH (P < 0.01). In conclusion, the protocol used for SARA induction was able to maintain ruminal pH between 5.5-6.0 for more than 48 hours. However, monensin and probiotics supplementation was not effective in preventing changes in ruminal and serum parameters during SARA.

The effect of pST on the testicular characteristics and metabolic parameters of prepubertal pigs was evaluated. Experiment 1 aimed to determine the interval between applications of pST based on the concentrations of circulating IGF-I. Experiment 2 aimed to evaluate the effect of pST on metabolic parameters, testicular characteristics, and expression of GHR, IGF-I and PCNA. In Experiment 1 twelve piglets with 30 days of age were used. The pST Group (n=6) was submitted to one i.m. injection of pST and the Control Group (n=6) to one placebo injection. Blood collections were performed until d 7 post pST application to determine IGF-I concentration and metabolic profile. In Experiment 2 twelve piglets with 22 days of age were used. The pST Group was submitted to pST injections every three days, and the Control Group received placebo doses during 30 days. Blood collections were performed every 3 days. Samples of liver and testicular tissue were collected to determine gene expression and testicular characteristics. In Experiment 1 IGF-I concentration was higher for the pST Group (P=0.02). In Experiment 2 the pST Group had higher body and testicular weight (P=0.06) and increased gene expression of PCNA in testes (P

The objective of this work was to evaluate the effect of insulin alone or in association with equine chorionic gonadotropin (eCG) on the fertility of postpartum beef cows subjected to synchronization. A total of 340 cows was subjected to fixed time artificial insemination. In the trial 1, the cows were subjected to temporary weaning (TW), while in the trial 2 the same protocol was tested without TW. The addition of an insulin injection to a progesterone/eCG-based protocol without TW increased the pregnancy rate of beef cows with body condition score (BCS) equal to or lower than 2.5. Insulin had no effect on cows submitted to TW or with BCS equal to or higher than 3.0.O objetivo deste trabalho foi avaliar apenas o efeito da insulina ou dela em associação com gonadotrofina coriônica equina (eCG) na fertilidade de vacas em pós-parto submetidas à sincronização. Um total de 340 vacas foi submetido à inseminação artificial em tempo fixo. No experimento 1, as vacas foram submetidas ao desmame temporário (DT), enquanto no experimento 2, o mesmo protocolo foi testado sem DT. A adição de uma injeção de insulina a um protocolo baseado em progesterona/eCG sem DT aumentou a taxa de prenhez de vacas de corte com escore de condição corporal (ECC) igual ou menor que 2,5. A insulina não teve efeito em vacas submetidas a DT ou com ECC igual ou maior que 3,0.

O objetivo deste estudo foi determinar o efeito do adsorvente (ADS) glucomanano modificado sobre parâmetros clínicos, hematológicos e ruminais de ovinos submetidos a dietas contendo aflatoxina (AFLA). Foram utilizadas 22 ovelhas mestiças (Corriedale e Texel), com 18 meses de idade. Os animais foram divididos em quatro grupos, de acordo com a adição de AFLA (1,5 PPM) ou ADS (2 PPM) na fração concentrada da dieta: animais recebendo AFLA na dieta (AFLA; n=6); animais recebendo AFLA e ADS na dieta (AFLA/ADS; n=6); animais recebendo ADS na dieta (ADS; n=5) e grupo controle, sem adição de ADS ou AFLA na dieta (CONTROLE; n=5). As dietas foram fornecidas por um período de 42 dias. Foram realizadas coletas de fluido ruminal a cada sete dias e avaliações clínicas (exame físico geral, urinálise e hemograma) a cada 10 dias. Quanto ao perfil leucocitário, a suplementação com adsorvente aumentou a concentração plasmática de eosinófilos. Os demais parâmetros clínicos e ruminais não foram influenciados pela presença do adsorvente ou da aflatoxina na dieta. Assim, o adsorvente glucomanano modificado influenciou a resposta leucocitária de ovinos, porém sem efeito na função ruminal e nos parâmetros clínicos avaliados.

O objetivo deste estudo foi avaliar o emprego de um análogo da prostaglandina F2α (PGF2α) em associação ao benzoato de estradiol (BE) em programas de sincronização de cios. Utilizaram-se 185 fêmeas não-lactantes divididas em quatro grupos. O G1 (44) e o G2 (46) receberam duas aplicações de PGF2α (D0 e D11) e os animais do G3 (26) e G4 (37) uma aplicação de PGF2α, após cinco dias de detecção de cio. O G1 e o G3 receberam 1mg de BE 48 horas após a PGF2α. A taxa de prenhez na IA foi de 42,2% no G1, 47,8% no G2, 32,1% no G3 e 57,6% no G4 (p>0,05) e de 38,4% e 51,4% para animais submetidos à IATF e à detecção de cio, respectivamente (p>0,05). Notou-se que a taxa de prenhez nas fêmeas inseminadas em tempo-fixo não diferiu daquelas que foram inseminadas após a detecção de cio. Palavras-chaves: Bovinos de corte, inseminação artificial em tempo fixo, prostaglandina, sincronização de cios.

O objetivo deste experimento foi avaliar o efeito das técnicas transcervical e laparoscópica sobre a taxa de prenhez de ovelhas inseminadas em tempo-fixo. A taxa de prenhez para as técnicas transcervical e laparoscópica foi de 40% (p>0.05). Quanto ao local de deposição do sêmen na inseminação artificial (IA) transcervical, não houve diferença na taxa de prenhez (p>0.05), com taxas de 25.0% na cervical superficial, 43.7% cervical médio, 41.7% cervical profundo e 50.0% intra-uterino. Estes resultados demonstram que a IA transcervical apresenta resultados semelhantes aos obtidos com o uso de laparoscopia, quando estas técnicas são realizadas em tempo-fixo. PALAVRAS-CHAVE: Cérvix, intrauterino, sêmen congelado, tracionamento