MicroRNAs as serum biomarker for Senecio brasiliensis poisoning in cattle.

Senecio spp. is one of the most frequent plant-related poisonings in cattle. Its ingestion generates the disease seneciosis, characterized by hepatic damages. Liver biopsies and serum markers dosage are tools used in diagnosis; however, many breeding cattle are undiagnosed. MicroRNAs are non-coding RNA, stable in biological fluids. Their difference in expression levels may indicate the presence of the poisoning. We analyzed the miRNA profiling to identify potential diagnostic biomarkers for Senecio brasiliensis poisoning. The expression of miR-21, miR-885, miR-122, miR-181b, miR-30a, miR-378, and let-7 f were evaluated in the serum of exposed cattle. At least one histological change was found in liver and lower quantity of albumin and high AST and ALP were also detected. MiRNAs miR-30a, miR-378, miR-21, miR-885, and miR-122 presented significantly higher expression in intoxicated animals than in healthy animals. Furthermore, miR-122, miR-885, and, especially, miR-21 signatures demonstrated high sensitivity and specificity, with potential application for detecting poisoning.

Novos valores de referência veterinários para volume plaquetário médio (MVP), amplitude de distribuição plaquetária (PDW) e plaquetócrito (PCT) na microrregião de Curitibanos / New veterinary reference values for mean platelet volume (MVP), platelet distribution width (PDW) and platelet count (PCT) in the Curitibanos microregion

Objetivo: O objetivo desse estudo foi elaborar os valores de referência desses parâmetros, baseado na casuística do Laboratório de Análises Clínicas Veterinárias (LaClin) da Universidade Federal de Santa Catarina para a microrregião de Curitibanos. Métodos: Avaliaram-se os novos índices plaquetários MPV, PDW e PCT de 100 caninos (41 machos e 59 fêmeas) e 40 felinos (13 machos e 27 fêmeas) saudáveis, atendidos no período de junho de 2015 até agosto de 2017 na faculdade de Medicina Veterinária da UFSC, na microrregião de Curitibanos, Santa Catarina. O valor de referência para MPV e PDW foi elaborado de acordo com a média dos valores obtidos dos animais avaliados ± 2 desvios-padrão, e, para PCT utilizou-se ± 1 DP. Resultados: Os valores de referência de MPV(fL), PDW(%) e PCT(%) [média (limite inferior ­ limite superior)] propostos para caninos foram, respectivamente 9,08 (7,44 -10,71), 16,07 (15,32 -16,82) e 0,32 (0,21-0,42), e para felinos, respectivamente 9,90 (8,07-11,74), 16,26 (15,21-17,32) e 0,24 (0,11-0,37). Conclusão: Os valores encontrados no presente estudo não condizem com outros estudos ou nunca foram investigados.

Comparação entre três métodos para detecção de glicosúria em cães e gatos / Comparison between three methods for detection of glycosuria in dogs and cats

Objetivo: Objetivou-se nesse estudo avaliar a presença de glicose na urina utilizando três metodologias. Métodos: Foram utilizadas 22 amostras de urina da rotina laboratorial, provenientes de 12 caninos e 10 felinos. As amostras foram submetidas a avaliação da glicosúria através de tiras reagentes urinárias ­ uma com leitura manual e outra automática ­ e reação de Benedict. Aplicou-se o teste de correlação de Pearson para se compararem os resultados entre os métodos. Resultados: A correlação entre os dois tipos de leituras de tiras reagentes foi alta (R=0,80), indicando que os resultados foram semelhantes entre as tiras, e baixa correlação entre ambas as tiras e o teste de Benedict (manual: R=0,35; automática: R=0,44), indicando que os resultados obtidos entre as tiras e o teste de Benedict não foram semelhantes. Conclusão: Os resultados permitem concluir que, assim como em humanos, o teste de Benedict é mais eficiente do que as tiras reagentes na detecção de glicosúria em cães e gatos, já que detectou um maior número de animais com glicosúria do que as tiras reagentes.

Effect of medroxy-progesterone acetate on follicular growth and endometrial cycloxygenase-2 (COX-2) expression during the bovine estrous cycle / Efeito do acetato de medroxi-progesterona sobre o crescimento filicular e expressão endometrial de ciclooxigenase-2 (COX-2) durante o ciclo estral de bovinos

The objective of this study was to evaluate the effect of medroxy-progesterone acetate (MAP) with or without estradiol benzoate (EB) on follicular growth during the estrous cycle in cattle. In the first experiment, Hereford cows were synchronized with a synthetic analogue of PGF2 alpha and were treated with two different doses of MAP (250 or 500 mg) with or without EB for 7 days starting on day 8 of the estrous cycle. Follicular growth was inhibited (P<0.05) in all cows except controls and those receiving 250mg MAP without EB. Seventy-five percent of the animals (15/20) showed estrus on days 21 and 22 of the cycle rather than at MAP withdrawal, demonstrating that these treatments did not induce estrus. To determine whether the EB treatment altered endometrial sensitivity to oxytocin and thus the luteolytic cascade, multiparous pre-synchronized cows received 5 mg of EB followed 6 hours later with 50 IU of oxytocin (OT; n=9). Eight hours after EB injection, endometrial fragments were collected from the cows on days 4, 13 and 17 of the estrous cycle and COX-2 gene expression was measured by PCR. EB increased COX-2 mRNA levels only on day 17 of the estrous cycle (P<0.05). In conclusion, MAP alone or associated with EB is able to suppress bovine follicular growth. However, EB in the presence of MAP is not efficient to induce luteolysis in cows when injected on day 8 of the estrous cycle.

Este estudo teve como objetivo avaliar o efeito do acetato de medroxi-progesterona (MAP) com ou sem benzoato de estradiol (BE) sobre o crescimento folicular durante o ciclo estral bovino. No primeiro experimento, vacas da raça Hereford foram sincronizadas com um análogo sintético de PGF2á e tratadas com duas doses diferentes de MAP (250 ou 500mg), com ou sem EB, durante 7 dias, iniciando-se no oitavo dia do ciclo estral. Observou-se uma inibição do crescimento folicular (P<0,05) em todas as vacas, exceto no grupo controle e no grupo que recebeu 250mg de MAP sem BE. Os 75 por cento dos animais não exibiu estro no momento da remoção do MAP, mas sim nos dias 21 e 22 do ciclo, demonstrando que os tratamentos não induziram cio. Para se determinar se o tratamento com BE alterou a sensibilidade endometrial à ocitocina e, assim, a cascata luteolítica, vacas multíparas pré-sincronizadas receberam 5mg de BE, seguidos, após 6 horas, de 50 UI de ocitocina (OT; n=9). Oito horas após a administração de BE, colheram-se fragmentos endometriais das vacas, nos dias 4, 13 e 17 do ciclo estral, mensurando-se a expressão gênica de COX-2 através de PCR. O BE aumentou os níveis de RNAm de COX-2 apenas no dia 17 do ciclo estral (P<0,05). Em conclusão, o MAP isolado ou associado a BE é capaz de suprimir o crescimento folicular bovino. Entretanto, o BE, na presença de MAP é ineficaz na indução da luteólise bovina, quando injetado no oitavo dia do ciclo estral.

Regulation of MMP2 and MMP9 metalloproteinases by FSH and growth factors in bovine granulosa cells.

Matrix metalloproteinases (MMP) are key enzymes involved in tissue remodeling. Within the ovary, they are believed to play a major role in ovulation, and have been linked to follicle atresia. To gain insight into the regulation of MMPs, we measured the effect of hormones and growth factors on MMP2 and MMP9 mRNA levels in non-luteinizing granulosa cells in serum-free culture. FSH and IGF1 both stimulated estradiol secretion and inhibited MMP2 and MMP9 mRNA abundance. In contrast, EGF and FGF2 both inhibited estradiol secretion but had no effect on MMP expression. At physiological doses, none of these hormones altered the proportion of dead cells. Although we cannot link MMP expression with apoptosis, the specific down regulation by the gonadotropic hormones FSH and IGF1 in vitro suggests that excess MMP2 and MMP9 expression is neither required nor desired for follicle development.

Regulation of MMP2 and MMP9 metalloproteinases by FSH and growth factors in bovine granulosa cells

Matrix metalloproteinases (MMP) are key enzymes involved in tissue remodeling. Within the ovary, they are believed to play a major role in ovulation, and have been linked to follicle atresia. To gain insight into the regulation of MMPs, we measured the effect of hormones and growth factors on MMP2 and MMP9 mRNA levels in non-luteinizing granulosa cells in serum-free culture. FSH and IGF1 both stimulated estradiol secretion and inhibited MMP2 and MMP9 mRNA abundance. In contrast, EGF and FGF2 both inhibited estradiol secretion but had no effect on MMP expression. At physiological doses, none of these hormones altered the proportion of dead cells. Although we cannot link MMP expression with apoptosis, the specific down regulation by the gonadotropic hormones FSH and IGF1 in vitro suggests that excess MMP2 and MMP9 expression is neither required nor desired for follicle development.

Regulation of angiotensin type 2 receptor in bovine granulosa cells.

Angiotensin II (AngII) is best known for its role in blood pressure regulation, but it also has documented actions in the reproductive system. There are two AngII receptors, type 1 (AGTR1) and type 2 (AGTR2). AGTR2 mediates the noncardiovascular effects of AngII and is expressed in the granulosa cell layer in rodents and is associated with follicle atresia. In contrast, expression of AGTR2 is reported to occur only in theca cells in cattle. The objective of the present study was to determine whether AngII also plays a role in follicle atresia in cattle. RT-PCR demonstrated AGTR2 mRNA in both granulosa and theca cells of bovine follicles. The presence of AGTR2 protein was confirmed by immunofluorescence. Abundance of AGTR2 mRNA in granulosa cells was higher in healthy compared with atretic follicles, whereas in theca cells, it did not change. Granulosa cells were cultured in serum-free medium, and treatment with hormones that increase estradiol secretion (FSH, IGF-I, and bone morphogenetic protein-7) increased AGTR2 mRNA and protein levels, whereas fibroblast growth factors inhibited estradiol secretion and AGTR2 protein levels. The addition of AngII or an AGTR2-specific agonist to granulosa cells in culture did not affect estradiol secretion or cell proliferation but inhibited abundance of mRNA encoding serine protease inhibitor E2, a protein involved in tissue remodeling. Because estradiol secretion is a major marker of nonatretic granulosa cells, these data suggest that AngII is not associated with follicle atresia in cattle but may have other specific roles during follicle growth.

Association of canine obesity with reduced serum levels of C-reactive protein.

The prevalence of obesity is increasing in dogs as well as in humans. C-reactive protein (CRP) is an important tool for the detection of inflammation and/or early tissue damage and is linked to obesity in humans. The objective of the present study was to determine if serum CRP levels are altered in obese dogs. Fifteen lean (control group) and 16 overweight (obese group) dogs were examined. Blood samples were collected under fasted conditions for serum determination of CRP, glucose, insulin, cholesterol, triglyceride, and fructosamine. Results indicated that obese dogs were insulin resistant because serum insulin and insulin/glucose ratios were higher than in lean dogs (P < or = 0.05). Serum CRP concentrations were lower in obese dogs than in controls (P < or = 0.001). C-reactive protein was negatively correlated with insulin/glucose ratio (R = -0.42) and cholesterol (R = -0.39; P < or = 0.05). Furthermore, levels of cholesterol, triglycerides, and fructosamine were increased in the obese group compared with the control group. Based on these results, it can be postulated that CRP production is inhibited by obesity and insulin resistance in dogs.

Ureteroneocistostomia extravesical modificada pela sondagem ureterovesical peroperatória no autotransplante renal em cães / Extravesical ureteroneocystostomy modified by the ureterovesical probe peroperative in kidney autotransplantation in dogs

Realizou-se, em sete cães adultos, o auto transplante renal esquerdo associado à nefrectomia contralateral para avaliação da técnica de ureteroneocistostomia extravesical modificada pela sondagem ureterovesical peroperatória. Durante a sutura do ureter na bexiga, foi mantida uma sonda uretral na região da anastomose ureterovesical, o que facilitou a realização da técnica cirúrgica e permitiu a confecção de anastomose de diâmetro adequado. A avaliação do rim transplantado e do ureter correspondente foi feita mediante ultra-sonografia a cada sete dias durante as seis primeiras semanas de pós-operatório, período em que também se fez o exame clínico diário de todos os animais. No último dia (42°) da avaliação a curto prazo, realizou-se a urografia excretora. Em seguida, manteve-se acompanhamento clínico periódico dos sete cães por um período mínimo de quatro meses, sem que fosse detectada qualquer alteração digna de nota. A técnica de reconstrução do trato urinário utilizada mostrou-se eficiente, não se observando nenhuma complicação urológica no pós-operatório de todos os animais.

To test a modified extravesical ureteroneocystostomy using a urethral probe like a stent just on the peroperative time, seven adult mongrel dogs underwent bilateral nephrectomy followed by unilateral autotransplantation. The other kidney was discarded. The evaluation of the animals was made by ultrasonography and clinical exams for the first six weeks after the surgery. The excretory urography was made on the end of this period. Then, the clinical exams were done by at least four months in each dog, without any complication. The technique of urinary tract reconstruction was considered efficient, without urological complications on the postoperative time of all the dogs.