Genetic polymorphisms in the angiotensin converting enzyme, actinin 3 and paraoxonase 1 genes in women with diabetes and hypertension

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.

Factors associated to serum paraoxonase 1 activity in patients with cardiovascular disease

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<0.05). Intake of cholesterol, saturated fatty acids (SFA) and monounsaturated fatty acids (MUFAS) was higher in patients with lower PON1 activity. Lipid ingestion tended to be higher in patients with lower PON1 activity (p=0.08). In the multivariate logistic regression model, SFA intake (P=0.03), genotype (P=0.09), gender (P=0.04), age (P=0.07) and carbohydrate intake (P=0.16) contributed the most to the serum PON1 activity. Conclusion: Based on these findings, nutritional guidance for these patients becomes essential, since dietary components interact with serum PON1 activity more than genotype.

The effect of paraoxonase 1 (PON1) gene polymorphisms T(-107)C and L55M and diet composition on serum PON1 activity in women

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.

The role of prostaglandin F2α on ovulation and LH release in cows / A função da prostaglandina F2α na ovulação e na liberação de LH em vacas

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 <0.0001) in cows from the GnRH group than in the PGF and Control groups. In experiment 4, cows with preovulatory follicles (>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.(AU)

O objetivo deste estudo foi avaliar o papel da prostaglandina F2α (PGF) na ovulação. No Experimento 1, as vacas foram alocadas aleatoriamente em dois tratamentos para receber 150 µg de d-Cloprostenol (Grupo PGF, n = 12) ou 2 mL de NaCl 0,9% (Grupo Controle, n = 11) e os CIDR, foram removidos 4 dias depois. Nenhuma vaca ovulou nos grupos Controle e PGF. No Experimento 2, as vacas foram separadas aleatoriamente em dois grupos experimentais para receber 4 injeções de 150 µg de d-Cloprostenol (n = 9) ou 2 mL de NaCL 0,9% (n = 9). Não foi observada ovulação em nenhum dos animais deste experimento. No Experimento 3, vacas ovariectomizadas receberam três injeções de 300µg de análogo de PGF (Grupo PGF, n = 5), 100µg de Lecirelina (Grupo GnRH, n = 5) ou 2 mL de PBS (Grupo Controle, n = 4). A concentração de LH foi maior (P <0,0001) nas vacas do grupo GnRH do que nos grupos PGF e Controle. No Experimento 4, vacas com folículos pré-ovulatórios (> 11,5 mm) foram tratadas com solução salina (Grupo Controle, n = 6), Lecirelina (Grupo GnRH, n = 7) ou Cloprostenol Sódico (Grupo PGF, n = 6). Houve um aumento significativo na área vascular dos folículos de 0 a 24h nos tratamentos com GnRH e PGF. Em conclusão, a PGF não foi capaz de induzir ovulação em vacas com alta ou baixa concentração plasmática de progesterona. Além disso, a PGF sozinha não foi capaz de induzir a liberação de LH e a luteinização do folículo, mas aumentou a vascularização folicular.(AU)

Prostaglandin E2 induces ovulation in prepubertal mice / Prostaglandina E2 induz ovulação em camundongos pré-púberes

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). Ep2gene 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.(AU)

O objetivo deste estudo foi determinar a capacidade da prostaglandina E2 (PGE2) em induzir a ovulação e expressão do receptor PGE2 (EP2 e EP4) e genes COX (COX-1 e COX-2) no ovário e na hipófise de camundongos pré-púberes. O controle positivo consistiu na aplicação de 5 µg de hormônio liberador de gonadotrofina (GnRH, n = 29); o controle negativo aplicação 0,5 mL de tampão fosfato-salino (PBS, n=31); o tratamento testado aplicação de 250 µg de PGE2 (n = 29), perfazendo um total de 89 camundongos (BALB/c) pré-púberes. Os camundongos foram sacrificados 14 a 15 h após os tratamentos para detectar ovulações e coleta de tecido. O teste do qui-quadrado foi usado para comparar a proporção de animais ovulando. As expressões gênicas e o número de ovulação foram analisados por ANOVA e o teste de tukey foi usado para comparar as médias entre os grupos. Uma maior proporção de camundongos (P <0,001) ovulou após receber GnRH (89,7%, 26/29) em comparação com o grupo PGE2 (58,6%, 17/29). No entanto, a proporção foi maior em comparação com aqueles tratados com PBS (0%, 0/31). A expressão do gene Ep2 na hipófise foi duas vezes maior (P <0,05) no grupo PGE2 em comparação com os grupos PBS e GnRH. Além disso, a PGE2 estimulou a Cox1(2,7 vezes, P <0,05) enquanto o GnRH estimulou a expressão de Cox2 (6,5 vezes, P <0,05) na pituitária em comparação com o grupo PBS. Em conclusão, nossos resultados suportam a hipótese de que PGE2 é capaz de induzir ovulação em camundongos pré-púberes com aumento concomitante na expressão dos genes Ep2 e Cox1 na glândula pituitária.(AU)

Frequency of alleles associated with celiac disease in patients with autoimmune thyroid disease / Frequência dos alelos associados com a doença celíaca em portadores de doenças autoimunes da tireoide

ABSTRACT Objective To determine the frequency of Human leukocyte antigen alleles and to verify the association of the presence of these alleles with symptoms and other diseases related to celiac disease in patients with autoimmune thyroid diseases. Methods A questionnaire on the symptoms and diseases associated with celiac disease was applied. Genomic deoxyribonucleic acid was extracted by collecting cells from the oral mucosa. The alleles (DQA1*0501; DQB1*0201; DRB1*04) were identified by means of the polymerase chain reaction. Results A total of 110 patients with autoimmune thyroid diseases participated in this study. It was observed that 66.4% of the individuals carried at least one of the alleles assessed and that 58.2% of the individuals were positive for at least one of the DQ2 alleles (DQA1*0501; DQB1*0201) and out of these 18.2% were positive for both DQ2 alleles (DQA1*0501; DQB1*0201). With regard to DQ8 (DRB1*04), 21.8% of the studied population was positive for this allele and 3.6% was positive for both DQ2 (DQA1*0501; DQB1*0201) and DQ8 (DRB1*04). A significant association was found between the presence of the DRB1*04 allele and gastrointestinal symptoms (p=0.02). A significant association of the DRB1*04 allele with type 1 diabetes mellitus (p=0.02) was observed. Conclusion The genetic profiles most commonly associated with celiac disease, such as DQ2 (DQA1*0501; DQB1*0201) and DQ8 (DRB1*04) were around 20.0% prevalent in the studied population. These are risk haplotypes for celiac disease especially when symptoms and diseases related to celiac disease are present. Therefore, it is important to screen patients to investigate a potential diagnosis for celiac disease.

RESUMO Objetivo Determinar a frequência dos alelos do Human leukocyte antigen e verificar a associação da presença desses alelos com sintomas e outras doenças relacionados à doença celíaca em portadores de doenças autoimunes da tireoide. Métodos Aplicou-se um questionário relacionado aos sintomas e doenças associados à doença celíaca. O ácido desoxirribonucleico genômico foi extraído por meio da coleta das células da mucosa bucal. Os alelos (DQA1*0501; DQB1*0201; DRB1*04) foram identificados por meio da reação em cadeia da polimerase. Resultados Participaram deste estudo 110 portadores de doenças autoimunes da tireoide. Observou-se que 66,4% dos indivíduos carregavam pelo menos um dos alelos estudados e que 58,2% dos indivíduos eram positivos para pelo menos um dos alelos DQ2 (DQA1*0501; DQB1*0201) e destes 18,2% foram positivos para ambos alelos do DQ2(DQA1*0501; DQB1*0201). Com relação ao DQ8 (DRB1*04), 21,8% da população estudada eram positivos para esse alelo e 3,6% eram positivos tanto para o DQ2 (DQA1*0501; DQB1*0201) quanto para o DQ8 (DRB1*04). Foi encontrada associação significativa da presença do alelo DRB1*04 com os sintomas gastrointestinais (p=0,02). Houve associação significativa do alelo DRB1*04 com diabetes mellitus tipo 1 (p=0,02). Conclusão O perfil genético mais fortemente associado à doença celíaca, tais como DQ2 (DQA1*0501; DQB1*0201) e DQ8 (DRB1*04) estavam presentes em torno de 20,0% da população estudada, estes são haplótipos de risco para doença celíaca e principalmente na presença de sintomas e doenças relacionadas à doença celíaca. Sendo assim, é importante realizar o rastreamento para investigar um possível diagnóstico para doença celíaca.

Influence of nutritional factors and the PON1 C(-107)T polymorphism on paraoxonase-1 activity in childhood / Influência de fatores nutricionais e do polimorfismo PON1 C(-107)T sobre a atividade da Paraoxonase 1 na infância

Abstract 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.

Paraoxonase 1 serum activity in women: the effects of menopause, the C(-107)T polymorphism and food intake

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.

The effect of vitamin C supplementation on neutropenia induced by cyclophosphamide in mice / Efecto con la suplementación de vitamina C sobre la neutropenia inducida por ciclofosfamida en ratones

ABSTRACT Neutropenia is one of the adverse effects caused by the administration of chemotherapy drugs. The present study aimed at investigating the effect of vitamin C supplementation in a model of immunosuppression induced by cyclophosphamide in mice. Vitamin C supplementation (50 mg/kg/day) administered intraperitoneally (i.p.) for 7 consecutive days in adult Swiss albino female. Mice were divided into four groups (n= 8/group): 1. Control (only distilled water i.p.), 2. Cyclophosphamide group (cyclophosphamide i.p. 150 and 100 mg/kg on days 1 and 4, respectively and distilled water daily), 3. Vitamin C group (Vitamin C 50 mg/kg i.p. and distilled water daily), and 4. Cyclophosphamide and Vitamin C group (cyclophosphamide i.p. 150 and 100 mg/kg on days 1 and 4, respectively and vitamin C 50 mg/kg i.p. daily). Vitamin C did not interfere in leukocytes count, but when co-administered with cyclophosphamide, significant interaction was observed, intensifying the neutropenia caused by cyclophosphamide. Vitamin C did not influence body weight during treatment, but groups receiving cyclophosphamide had a significant weight loss from the third day of treatment until the end of experiment compared to the control group. Vitamin C supplementation intensified neutropenia induced by cyclophosphamide and did not prevent weight loss induced by cyclophosphamide in mice.

RESUMEN La neutropenia es uno de los efectos adversos causados por la administración de medicamentos de quimioterapia. El presente estudio tuvo como objetivo investigar el efecto de la administración de suplementos de vitamina C en un modelo de inmunosupresión inducida por ciclofosfamida en ratones. Se realizó la suplementación de vitamina C (50 mg/kg/día) administrada por vía intraperitoneal (i.p) por 7 días en ratones hembras swiss albinas adultas. Los ratones fueron divididos en cuatro grupos (n= 8 / grupo): Control (solamente agua destilada i.p) Ciclofosfamida (ciclofosfamida i.p. 150 mg/kg y 100 mg/kg en el día 1 y día 4 y agua destilada diariamente) Vitamin C (vitamina C 50 mg/kg i.p y agua destilada diariamente), Ciclofosfamida y Vit C (ciclofosfamida i.p. 150 y 100 mg/kg en el día 1 y día 4 y vitamina C 50 mg/kg i.p. diariamente). La vitamina C por sí no interfirió en los valores de leucocitos y tampoco influyó en el peso corporal durante el tratamiento, pero los grupos que recibieron ciclofosfamida tuvieron una pérdida de peso significativa desde el tercer día de tratamiento hasta el final del experimento en comparación con el grupo control. La suplementación de vitamina C intensificó la neutropenia inducida por ciclofosfamida y no evitó la pérdida de peso inducida por ciclofosfamida en ratones.

Evaluation of the Redox State of Wistar Rats Submitted to High-Fat Diet Supplemented With Infusion of Ilex paraguariensis

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.

Systemic inflammatory and stress markers in cattle and sheep submitted to different reproductive procedures / Marcadores sistêmicos de inflamação e estresse em bovinos e ovinos submetidos a diferentes procedimentos reprodutivos

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.

RESUMO: Estudos anteriores avaliaram o efeito de diferentes procedimentos reprodutivos sobre marcadores de desconforto em bovinos e ovinos. Tais estudos podem estimular a adoção de técnicas que preservem o bem-estar animal. Entretanto, os marcadores comumente utilizados apresentam alta influência de fatores externos. Para contornar isso, a avaliação conjunta de diferentes parâmetros sistêmicos pode ser utilizada para determinar com maior precisão a presença de estresse, dor ou inflamação, como cortisol, proteínas de fase aguda, a bradicinina e a substância P. O objetivo desta revisão é relacionar potenciais marcadores de inflamação e estresse, discutindo como e quando são regulados frente aos estímulos em bovinos e ovinos. Ainda, pretende-se revisar de que forma procedimentos reprodutivos com diferentes graus de invasividade acarretam em desconforto, fornecendo informações para a elaboração de estratégias que possibilitem minimizá-lo.

Seroprevalence estimate and associated risk factors for neosporosis in dairy cattle in the northwest region of Rio Grande do Sul State, Brazil / Estimativa da soroprevalência e fatores de risco para neosporose em rebanhos leiteiros na região Noroeste do estado do Rio Grande do Sul, Brasil

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.

RESUMO: O objetivo desse estudo foi estimar a soroprevalência da neosporose e os possíveis fatores de risco em rebanhos (Bos taurus taurus) localizados na mesorregião Noroeste do Rio Grande do Sul, Brasil. Foram coletadas 322 amostras de sangue de bovinos leiteiros, em 18 propriedades localizadas em 17 munícipios desta mesorregião. Um questionário epidemiológico foi aplicado em cada propriedade, contendo questões relacionadas às características gerais dos rebanhos, dados reprodutivos e manejo animal. As amostras de soro foram testadas através do teste de imunoensaio enzimático (ELISA) para Neospora caninum. Os resultados demonstraram uma soroprevalência de Neospora de 88,9% (16/18) entre os rebanhos e 31,1% (100/322) entre os indivíduos. Entre os fatores de risco analisados foi observado que descarte por problemas reprodutivos (OR=0,6), presença de áreas alagadiças (OR=0,5) e venda comercial (OR=0,4) estavam associados a soroprevalência. No entanto, a compra de animais substituídos no rebanho desempenhou um papel significativo na ocorrência da doença (OR=2,2). Os resultados desse estudo sugerem que o Neospora caninum esteve presente nos rebanhos estudados, bem como, existem fatores associados com a soroprevalência nas propriedades da mesorregião do Noroeste do Rio Grande do Sul.

Association between paraoxonase 1 (PON1) enzyme activity, PON1 C(−107)T polymorphism, nutritional status, and lipid profile in children

BACKGROUND: Paraoxonase 1 (PON1) is an enzyme that possesses anti-atherogenic and anti-inflammatory properties with serum levels determined by genetic and exogenous factors. Lower serum PON1 arylesterase activity is associated to metabolic alterations related to childhood overweight and onset and/or development of diabetes and CVD later in life. However, data on the relationship between genetic PON1 polymorphisms and nutritional status as well as lipid profile in children are limited. To investigate the distribution of the C(−107)T PON1 gene polymorphism and its relation with serum PON1 enzyme activity, nutritional status and lipid profile in children. METHODS: A cross-sectional study was performed including 73 children aged 5 to 7 years who attended public pediatric clinics. PON1 C(−107)T, arylesterase activity, body mass index for the age, and serum lipid profile were evaluated. RESULTS: PON1 activity was higher in overweight children compared to the normal weight ones (p= 0.02). The genotypic frequency did not differ between the two groups (p> 0.05). Carriers of CC genotype had higher enzyme activity than T allele carriers, and this difference was greater among normal weight children. HDL levels were higher among normal weight children carrying CC genotype, compared to those carrying the T allele (p< 0.01).CONCLUSION: The PON1 C(−107)T polymorphism is associated with higher serum enzyme activity in children, as observed previously in adults. In addition, this polymorphism also shows association to higher high density lipoprotein (HDL) levels and serum PON1 arylesterase activity in the normal weight children studied.

Age at onset of caloric restriction and its effects on the redox profile of the rat hippocampus

BACKGROUND: The benefits of caloric restriction (CR) on the protection against age-related neurodegenerative diseases have been the subject of several studies. However, the effects of CR on the central nervous system are still poorl y understood since most studies were carried out in mature animals. The present study aimed to investigate whether the age at onset of CR could differently affect the redox status of the rat hippocampus. METHODS: Thirty-two male Wistar rats at 35 days old (35d;n= 16) and 65 days old (65d;n= 16) were fed ad libitum or subjected to 30 % CR (n= 8 group/age) for 12 weeks. At the end of the experiment, the rats were euthanized, blood was collected, and the hippocampus was dissected for measuring the redox status. RESULTS: CR in 35d and 65d rats induced a 16 and 21% reduction in body weight gain, respectively, compared to controls (p< 0.05). Urea, total cholesterol, triacylglycerol, HDL cholesterol, and LDL cholesterol concentrations were lower in CR 35d rats than in 35d controls (p< 0.05). No differences were detected between the CR groups and controls in the object recognition test (p> 0.05) and in superoxide dismutase activity, nitric oxide content, and lipid peroxidation levels(p> 0.05). However, glutathione peroxidase activity was higher (p< 0.0001) in 65d rats compared to that in 35d rats, and GSH content was higher (p< 0.05) in CR-fed rats compared to that in controls at both ages. CONCLUSIONS: In conclusion, CR increased GSH content when started at both ages but did not affect the activity of antioxidant enzymes and the level of ROS in the hippocampus. In addition, CR did not induce any detrimental effects on memory and nutritional status when started in both 35d and 65d rats

Impacto da suplementação de vitamina C sobre níveis de peroxidação lipídica e glutationa reduzida em tecido hepático de camundongos com imunossupressão induzida por ciclofosfamida / Impact of vitamin C supplementation on lipid peroxidation and reduced glutathione levels in the liver tissue of mice with cyclophosphamide-induced immunosuppression

RESUMO Objetivo: Investigar os efeitos da vitamina C sobre níveis de peroxidação lipídica e glutationa reduzida em tecido hepático de camundongos imunossuprimidos por ciclofosfamida. Métodos: O estudo foi realizado em camundongos Swiss, fêmeas, com 45 dias de idade, separados em quatro grupos com oito animais cada. Grupos: controle (água destilada), vitamina C (50 mg/kg), ciclofosfamida (100 + 150 mg/kg) e tratamento (vitamina C 50 mg/kg + ciclofosfamida 100 +150 mg/kg). Todas as aplicações foram via intraperitoneal. O ensaio biológico teve duração de seis dias, sendo o sétimo a eutanásia dos animais. As análises bioquímicas de peroxidação lipídica (quantificação de substâncias reativas ao ácido tiobarbitúrico) e glutationa reduzida (estimativa de tiois não proteicos) foram realizadas em tecido hepático. Resultados: A ciclofosfamida causou aumento significativo (p<0,0001) nos níveis de peroxidação lipídica. Não foram observadas alterações significativas nos grupos tratados com vitamina C. A ciclofosfamida por si só, não alterou níveis de glutationa reduzida. A vitamina C causou a redução do nível de glutationa reduzida em relação ao controle tanto nos animais que receberam ciclofosfamida quanto nos que não receberam. No entanto, nos grupos tratados com o quimioterápico houve uma interação entre a droga e a vitamina, ou seja, o quimioterápico intensificou a diminuição da glutationa reduzida provocada pela vitamina C. Conclusão: A ciclofosfamida, na dose e período utilizados, foi capaz de induzir o dano oxidativo verificado pelo aumento da peroxidação lipídica. A vitamina C, na dose de 50 mg/kg de peso, não apresentou potencial para proteger contra o dano oxidativo provocado pelo quimioterápico.

ABSTRACT Objective: To investigate the effects of vitamin C supplementation on the levels of lipid peroxidation and reduced glutathione in the liver tissue of mice immunosuppressed with cyclophosphamide. Methods: Thirty-two 45-day-old female Swiss mice were divided into four groups of eight animals each as follows: control (distilled water); vitamin C (50 mg/kg); cyclophosphamide (100 + 150 mg/kg); and treatment (vitamin C 50 mg/kg + cyclophosphamide 100 +150 mg/kg). The substances were provided intraperitoneally for six days, and on the seventh day, the mice were euthanized. The biochemical analyses of lipid peroxidation (quantification of thiobarbituric acid-reactive substances) and reduced glutathione (estimate of non-protein thiols) were performed on liver tissue. Results: Cyclophosphamide increased the levels of lipid peroxidation (p<0.0001). Significant changes were not found in the groups treated with vitamin C. Cyclophosphamide alone did not affect the levels of reduced glutathione. Compared with the control group, vitamin C reduced the levels of reduced glutathione in animals that received or not cyclophosphamide. Vitamin C interacted with cyclophosphamide, that is, the chemotherapeutic agent further decreased the lower levels of reduced glutathione secondary to vitamin C intake. Conclusion: Cyclophosphamide, in the study dosage and duration, was capable of inducing oxidative damage, verified by increased lipid peroxidation. A vitamin C dosage of 50mg/kg of body weight did not protect against the oxidative damage caused by the chemotherapeutic agent.

Effect of porcine somatotropin on metabolism and testicular characteristics of prepubertal pigs / Efeito da somatotrofina suína sobre o metabolismo e características testiculares de suínos pré-púberes

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 day 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 < 0.05). However, a reduction in the number of seminiferous tubules, and Sertoli cells, and in GHR expression (P < 0.05) was observed. Thus, pST administration increased body and testis development in prepubertal pigs, however it reduced the density of seminiferous tubules and Sertoli cells.

Foi investigado o efeito da pST sobre características testiculares e metabolismo de suínos pré-púberes. O Experimento 1 determinou o intervalo entre aplicações de pST, baseado nas concentrações de IGF-I. O Experimento 2 avaliou o efeito da pST sobre o metabolismo, características testiculares e expressão gênica de GHR, IGF-I e PCNA. No Experimento 1, foram usados 12 leitões com 30 dias de idade. O grupo pST (n = 6) foi submetido a uma injeção IM de pST e o grupo Controle (n = 6) a uma injeção de placebo. Coletas de sangue foram realizadas até o dia sete após a aplicação de pST para determinação dos níveis de IGF-I e parâmetros metabólicos. No Experimento 2, foram usados 12 leitões com 22 dias de idade. O grupo pST foi submetido às aplicações de pST a cada 3 dias, e o grupo Controle, às doses de placebo, durante 30 dias. Coletas de sangue foram realizadas a cada três dias. Amostras de fígado e testículo foram coletadas para determinar a expressão gênica e características testiculares. No Experimento 1, a concentração de IGF-I foi maior no grupo pST (P = 0,02). No Experimento 2, o grupo pST teve maior peso corporal e testicular (P = 0,06) e aumento na expressão de PCNA no testículo (P < 0,05). Contudo, foi observada uma redução no número de túbulos seminíferos, células de Sertoli e GHR (P < 0,05). Assim, a administração de pST aumentou o desenvolvimento testicular e corporal de suínos pré-púberes, porém reduziu a densidade de túbulos seminíferos e células de Sertoli.

The use of sodic monensin and probiotics for controlling subacute ruminal acidosis in sheep / O uso de monensina sódica e probióticos para o controle de acidose ruminal subaguda em ovinos

It was to validated 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.

Foi validado um protocolo para a indução de acidose ruminal subaguda (SARA) (Experimento 1) e testar a eficácia do probiótico Saccharomyces cerevisiae ou monensina na prevenção da queda do pH do fluido ruminal em ovinos (Experimento 2). No Experimento 1, seis ovelhas foram mantidas em jejum por dois dias e, em seguida, alimentadas basicamente com concentrado durante quatro dias. Nesse protocolo as ovelhas mantiveram o pH do fluido ruminal abaixo de 6,0 por 75 horas consecutivas. No Experimento 2, 18 ovelhas foram distribuídas em três grupos: controle (GC, n = 6), monensina (GM, n = 6) e o grupo probiótico (GP, n = 6). A SARA foi induzida de acordo com o Experimento 1. GP apresentaram valores de pH mais baixos (5,7 ± 0,1) do que o GC (6,0 ± 0,1) (P = 0,05), enquanto GM (5,7 ± 0,1) foi semelhante durante a indução de SARA. A indução SARA reduziu a população de protozoários no rúmen (P < 0,05) e aumentou a concentração de cloreto no líquido ruminal (P < 0,01). Durante a SARA observou-se aumento das concentrações séricas de fósforo (P < 0,01), AST (P < 0,01) e GGT (P < 0,01), mas reduziu a de LDH (P < 0,01). Em conclusão, o protocolo utilizado para a indução de SARA foi capaz de manter o pH do rúmen entre 5,5-6,0 por períodos superiores a 48 horas. No entanto, a suplementação com monensina e probióticos não foi eficaz na prevenção das alterações nos parâmetros ruminais e séricos durante SARA.

Efeito da adição de glucomanano modificado em dietas contendo aflatoxina sobre parâmetros clínicos de ovinos / Effect of the modified glucomannan in diets containing aflatoxin on the clinical parameters in sheep

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.

The aim of this study was to determine the effect of modified glucomannan adsorbent (ADS) on the clinical, hematological and ruminal parameters in sheep subjected to diets containing aflatoxins (AFLA). Twenty two crossbred ewes (Corriedale vs Texel), aging 18 months were separated into four groups according to the addition of AFLA (1,5 PPM) or ADS (2 PPM) in the concentrate portion of the diet: animals receiving AFLA in the diet (AFLA; n=6); animals receiving AFLA and ADS in the diet (AFLA/ADS; n=6); animals receiving ADS in the diet (n=5) and control group, without addition of the ADS or AFLA in the diet (CONTROL; n=5). The diets were supplied during 42 days. Ruminal fluid was collected and analysed every seven days and the clinical evaluations (clinical examination, urinalysis, and hemogram) every 10 days. The addition of adsorbent increased the plasmatic concentration of eosinophiles. Clinical and ruminal parameters were not affected by the presence of adsorbent or aflatoxin into the diet. The modified glucomannan adsorbent influenced the leukocyte response of sheep, but no effect on ruminal function and clinical parameters.

Efeito da somatotropina recombinante bovina (rbST) sobre os níveis de marcadores metabólicos de touros semiconfinados das raças Aberdeen Angus e Brangus / Effect of recombinant bovine somatotropin (rbST) on the concentrations of metabolic markers in semi-confined sires Aberdeen Angus and Brangus

O objetivo deste estudo foi avaliar o efeito da somatotropina recombinante bovina (rbST) sobre os marcadores do metabolismoenergético, proteico, mineral e hepático de touros mantidos em semiconfinamento. Foram utilizados 20 touros de raçasAberdeen Angus e Brangus, sendo divididos aleatoriamente em dois grupos (Placebo e rbST), para que no Placebo fosseadministrado via subcutânea duas doses de NaCl, 0,9% e no rbST fosse administrado o rbST (Lactotropin®). Foram realizadascinco coletas de sangue, com intervalo de 14 dias, para avaliação da concentração de triacilgliceróis e colesterol comomarcadores energéticos, ureia, como indicador proteico, fosfatase alcalina e fósforo como marcadores minerais e asenzimas aspartato aminotransferase (AST) e a gama glutamiltransferase (GGT) como indicadoras da função hepática. Aaplicação de rbST não influenciou os marcadores energéticos, minerais, e proteico, exceto a enzima hepática GGT (P 0,05).Estes resultados indicam que a utilização do rbST em touros não alterou o metabolismo, pois os parâmetros avaliadosmantiveram-se dentro dos padrões considerados fisiológicos para a espécie.

The aim of this study was to evaluate the effect of recombinant bovine somatotropin (rbST) on metabolic markers for energy, protein, mineral and hepatic metabolism of sires kept grazing and receiving concentrate. Twenty sires Aberdeen Angus and Brangus used for this experiment were divided in two groups (Placebo and rbST). The group Placebo received subcutaneous administration of NaCl, 0.9% and rbST receive rbST twice. Five blood collections with 14 days of interval were made for evaluation of triglyceride and cholesterol as energetic metabolism markers, urea, as proteic marker, alkaline phosphatase and phosphorus as minerals markers and aspartate aminotransferase (AST) and gamma glutamyltransferase (GGT) as indicators of the hepatic function. The rbST did not influence the energetic, mineral and proteic markers, except the hepatic enzyme GGT (P<0,05). These results indicated that the use of rbST in sires did not affect metabolism, therefore evaluated parameters remaining in the physiological range for the specie.

Factors that affect the in vitro production of bovine embryos. A review / Factores que afectan la producción in vitro de embriones bovinos: una revisión / Fatores que afetam a produção in vitro de embriões bovinos. A revisão

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.