Identificação imuno-histoquímica de VEGF e IGF-1 em ovários de cadelas no anestro e estro / Immunohistochemical identification of VEGF and IGF-1 in ovaries of bitches on anestrus and estrus

Com o objetivo de verificar a presença de VEGF e IGF-1 nos ovários de cadelas, foram realizadas análises imuno-histoquímicas do estroma cortical; teca e granulosa de folículos secundários, terciários e terciários pré-ovulatórios luteinizados; e ovócitos de folículos primários, secundários e terciários de ovários de cinco cadelas em anestro (Anest) e cinco em estro (Est). A identificação das fases do ciclo estral foi realizada por citologia vaginal associada a dosagem plasmática de progesterona. Os ovários foram submetidos a tratamento imuno-histoquímico para identificação de VEGF (anticorpo primário PU 360-UP, Biogenex, USA; diluição 1:30) e IGF-1 (anticorpo primário PabCa, Gro-Pep, Austrália; diluição 1:100). Determinou-se um índice de imunomarcação (IM), para cada tecido avaliado, pela razão entre a área positivamente marcada dividida pela área total analisada. Para os ovócitos, verificou-se imunomarcação positiva ou negativa. As comparações de IM entre tecidos foram realizadas pelo teste de Wilcoxon (diferentes tecidos em mesmo grupo) ou Mann-Whitney (mesmo tecido entre diferentes grupos), todas no nível de 5% de significância. VEGF e IGF-1 foram identificados, de forma semelhante (P>0,05), em todas as estruturas avaliadas em ambos os grupos experimentais. Conclui-se que esses fatores de crescimento estão presentes em cadelas no anestro e estro, no estroma cortical ovariano, folículos em diferentes estádios de desenvolvimento e ovócitos.(AU)

In order to verify the presence of VEGF and IGF-1 in the ovaries of bitches, immunohistochemical analyzes of the cortical stroma; theca and granulosa of secondary, tertiary and tertiary luteinized preovulatory follicles; and oocytes of primary, secondary and tertiary follicles of ovaries from five bitches in anestrous (Anest) and five in estrus (Est) was performed. The identification of the phases of the estrous cycle was performed by vaginal cytology associated with the measurement of plasma progesterone. The ovaries were treated for immunohistochemical identification of VEGF (PU 360 primary antibody-UP, Biogenex, USA, dilution 1:30) and IGF-1 (primary antibody PabCa, Gro-Pep, Australia; 1:100 dilution). The immunostaining index (MI) was determined for each tissue by the ratio of positively marked area divided by total analyzed area. For oocytes immunostaining was determined as positive or negative. Comparisons of IM between tissues were performed with the Wilcoxon test (deferent tissues in the same group) or Mann-Whitney test (same tissue between different groups), all at 5% significance level. VEGF and IGF-1 have been similarly identified (P>0.05) in all structures evaluated in both groups. It is concluded that in bitches in estrus and anestrous these growth factors are present in ovary cortical stroma, follicles at different stages of development and oocytes.(AU)

Identificação imuno-histoquímica de VEGF e IGF-1 em ovários de cadelas no anestro e estro / Immunohistochemical identification of VEGF and IGF-1 in ovaries of bitches on anestrus and estrus

Com o objetivo de verificar a presença de VEGF e IGF-1 nos ovários de cadelas, foram realizadas análises imuno-histoquímicas do estroma cortical; teca e granulosa de folículos secundários, terciários e terciários pré-ovulatórios luteinizados; e ovócitos de folículos primários, secundários e terciários de ovários de cinco cadelas em anestro (Anest) e cinco em estro (Est). A identificação das fases do ciclo estral foi realizada por citologia vaginal associada a dosagem plasmática de progesterona. Os ovários foram submetidos a tratamento imuno-histoquímico para identificação de VEGF (anticorpo primário PU 360-UP, Biogenex, USA; diluição 1:30) e IGF-1 (anticorpo primário PabCa, Gro-Pep, Austrália; diluição 1:100). Determinou-se um índice de imunomarcação (IM), para cada tecido avaliado, pela razão entre a área positivamente marcada dividida pela área total analisada. Para os ovócitos, verificou-se imunomarcação positiva ou negativa. As comparações de IM entre tecidos foram realizadas pelo teste de Wilcoxon (diferentes tecidos em mesmo grupo) ou Mann-Whitney (mesmo tecido entre diferentes grupos), todas no nível de 5% de significância. VEGF e IGF-1 foram identificados, de forma semelhante (P>0,05), em todas as estruturas avaliadas em ambos os grupos experimentais. Conclui-se que esses fatores de crescimento estão presentes em cadelas no anestro e estro, no estroma cortical ovariano, folículos em diferentes estádios de desenvolvimento e ovócitos.(AU)

In order to verify the presence of VEGF and IGF-1 in the ovaries of bitches, immunohistochemical analyzes of the cortical stroma; theca and granulosa of secondary, tertiary and tertiary luteinized preovulatory follicles; and oocytes of primary, secondary and tertiary follicles of ovaries from five bitches in anestrous (Anest) and five in estrus (Est) was performed. The identification of the phases of the estrous cycle was performed by vaginal cytology associated with the measurement of plasma progesterone. The ovaries were treated for immunohistochemical identification of VEGF (PU 360 primary antibody-UP, Biogenex, USA, dilution 1:30) and IGF-1 (primary antibody PabCa, Gro-Pep, Australia; 1:100 dilution). The immunostaining index (MI) was determined for each tissue by the ratio of positively marked area divided by total analyzed area. For oocytes immunostaining was determined as positive or negative. Comparisons of IM between tissues were performed with the Wilcoxon test (deferent tissues in the same group) or Mann-Whitney test (same tissue between different groups), all at 5% significance level. VEGF and IGF-1 have been similarly identified (P>0.05) in all structures evaluated in both groups. It is concluded that in bitches in estrus and anestrous these growth factors are present in ovary cortical stroma, follicles at different stages of development and oocytes.(AU)

Eosinophils and mast cells in the oviduct of heifers under natural and superovulated estrous cycles

Quantification of mucosal eosinophils and mast cells in the oviducts from 22 crossbred heifers was performed in both natural (estrus, metaestrus and diestrus) and superovulated (estrus and metaestrus) estrous cycles. The number of cells/square millimeter of mucosa was obtained through counts at different regions of the oviduct (infundibulum, ampulla, ampullary/isthmic transition and isthmus). Differences were not found (P > 0.05) among numbers of cells at the different phases of the natural estrous cycle nor between natural or superovulated cycles. When all animals were separated into two phases of the estrous cycle (estrus and metaestrus), the number of eosinophils at the ampullary/isthmic transition and isthmus was higher (P < 0.01) at the estrus cycle phase, and the number at the infundibulum was higher (P < 0.001) during the metaestrus phase than at estrus. The number of mast cells was highest at the isthmus regardless of the estrus phase, and highest at the infundibulum (P < 0.001) and ampulla (P < 0.05) during the metaestrus phase. Significant correlations were found between the number of mast cells and plasma progesterone levels at the infundibulum (P < 0.001; r = 0.69) and ampulla (P < 0.03; r = 0.51). No correlation was seen between numbers of eosinophils and mast cells, progesterone and 17β-estradiol concentrations. Therefore, the distribution of mucosal eosinophils in the oviduct of heifers is apparently not related to the circulating levels of 17β-estradiol and progesterone; however, the highest number of mast cells found at the metaestrus phase of the cycle, at least for the infundibulum and ampulla, appears to be related to the high circulating progesterone plasma levels in metaestrus phase compared to estrus phase. No differences were found between animals subjected or not to superovulation.(AU)

Eosinophils and mast cells in the oviduct of heifers under natural and superovulated estrous cycles

Quantification of mucosal eosinophils and mast cells in the oviducts from 22 crossbred heifers was performed in both natural (estrus, metaestrus and diestrus) and superovulated (estrus and metaestrus) estrous cycles. The number of cells/square millimeter of mucosa was obtained through counts at different regions of the oviduct (infundibulum, ampulla, ampullary/isthmic transition and isthmus). Differences were not found (P > 0.05) among numbers of cells at the different phases of the natural estrous cycle nor between natural or superovulated cycles. When all animals were separated into two phases of the estrous cycle (estrus and metaestrus), the number of eosinophils at the ampullary/isthmic transition and isthmus was higher (P < 0.01) at the estrus cycle phase, and the number at the infundibulum was higher (P < 0.001) during the metaestrus phase than at estrus. The number of mast cells was highest at the isthmus regardless of the estrus phase, and highest at the infundibulum (P < 0.001) and ampulla (P < 0.05) during the metaestrus phase. Significant correlations were found between the number of mast cells and plasma progesterone levels at the infundibulum (P < 0.001; r = 0.69) and ampulla (P < 0.03; r = 0.51). No correlation was seen between numbers of eosinophils and mast cells, progesterone and 17β-estradiol concentrations. Therefore, the distribution of mucosal eosinophils in the oviduct of heifers is apparently not related to the circulating levels of 17β-estradiol and progesterone; however, the highest number of mast cells found at the metaestrus phase of the cycle, at least for the infundibulum and ampulla, appears to be related to the high circulating progesterone plasma levels in metaestrus phase compared to estrus phase. No differences were found between animals subjected or not to superovulation.

Alterações morfológicas induzidas por butirato, propionato e lactato sobre a mucosa ruminal e epiderme de bezerros: II. Aspectos ultra-estruturais / Lactate, propionate, and butyrate induced morphological alterations on calf ruminal mucosa and epidermis: II. Ultra-structurals aspects

Avaliou-se o efeito de ácidos graxos voláteis (AGV) sobre a integridade do epitélio no rúmen, no plano nasolabial, na epicera e no perioplum traseiro e dianteiro de bezerros e validou-se a feitura de biópsias tegumentares como indicadores de alterações morfológicas da mucosa ruminal. Dezessete bezerros, com sonda no rúmen, receberam infusões intra-ruminais de AGV ou salina, durante 37 dias. Aos 89 dias de vida, após o abate, foram colhidas amostras dos tecidos. Os AGV aumentaram a área de epitélio total e a área de células metabolicamente ativas no epitélio ruminal, embora o butirato não tenha induzido ao desenvolvimento papilar. A área de epitélio não queratinizado no plano nasolabial foi reduzida pela infusão de AGV. Butirato e lactato foram mais indutores de alterações patológicas no epitélio ruminal. Não foram observadas lesões histológicas nos epitélios do plano nasolabial, da epicera e do perioplum, mostrando que essas são conseqüências do efeito direto dos AGV sobre o epitélio ruminal. Os efeitos indireto e direto dos AGV sobre a morfologia dos tecidos epiteliais queratinizados não foram iguais. Biópsias tegumentares podem ter utilidade como indicadores de alterações morfológicas da mucosa ruminal.(AU)

The effect of volatile fatty acids (VFA) on rumen wall, epidermis of nasolabial surface, perioplum, and epicera of calves was evaluated. The experiment also aimed to validate the procedure of tegument biopsies as indicators of ruminal mucosa alterations. Seventeen neonatal calves with foley catheters received intraruminal infusions of VFA or saline, during 37 days. At 89-day-old, the animals were slaughtered and tissue samples were collected from rumen, nasolabial surface, epicera, and perioplum from face and hindquarters. VFA infusion increased total epithelium area and metabolically active ruminal cell area; although butirate did not induce the papilar development. The effect of nasolabial surface VFA infusion was the opposite to that observed in the rumen. No histological lesion was observed on nasolabial surface, epicera, and perioplum, demonstrating that these are consequences of VFA direct effect on ruminal epithelium. Butyrate and lactate induced more alterations on the ruminal epithelium. Indirect and direct VFA effects on keratinized epithelium tissues morphology were not identical. Tegumentary biopsies may be useful as indicators of morphological alterations of ruminal mucosa.(AU)

Spindle cell sarcoma in Bothrops leucurus / Sarcoma fusocelular em jararaca - Bothrops leucurus

O presente relato descreve os achados anatomopatológicos de uma neoplasia maligna em jararaca (Bothrops leucurus), mantida em cativeiro durante sete anos. O animal apresentava massa nodular subcutânea localizada no lado direito do terço anterior. O exame histológico revelou tratar-se de neoplasia mesenquimal maligna, permitindo o diagnóstico de sarcoma fusocelular.(AU)