Morfologia e grânulos citoplasmáticos do corpo lúteo de vacas aneloradas (Bos taurus indicus) gestantes e não gestantes / Morphology and cytoplasmic granules of the corpus luteum in pregnant and non-pregnant Nelore cows

Avaliaram-se a proporção volumétrica dos constituintes do corpo lúteo e a distribuição de células lúteas com grânulos citoplasmáticos no corpo lúteo de animais Nelore ao longo da gestação e em animais não gestantes. Foram coletados ovários com corpos lúteos de 24 animais abatidos em frigorífico. Os animais foram distribuídos em dois grupos: gestantes e não gestantes. A idade gestacional foi determinada pela medição do comprimento apicocaudal do feto. Os ovários foram distribuídos segundo o trimestre gestacional: primeiro - gestação de até 90 dias; segundo - de 91 a 180 dias; e terceiro - de 181 a 270 dias. Os corpos lúteos, processados para inclusão em paraplast, foram avaliados em microscópio de luz. Na proporção volumétrica do corpo lúteo, os constituintes avaliados foram citoplasma e núcleo de células lúteas, tecido conjuntivo e fibroblastos, células endoteliais e pericitos e vasos sanguíneos. A proporção volumétrica de citoplasma de células lúteas reduziu de 46,1±2,8% para 37,9±3,5% e a de núcleo de células lúteas reduziu de 9,2±1,0% para 6,2±1,0%, do primeiro para o segundo trimestre. A proporção de tecido conjuntivo e fibroblastos aumentou de 20,9±5,4%, no primeiro trimestre, para 34,0±4,2%, no segundo trimestre. Grânulos citoplasmáticos foram evidenciados pelas técnicas histoquímicas xylidine ponceau e azul de bromofenol, que indicam sua provável composição proteica. A quantidade de células maiores com grânulos aumentou significativamente de 2,3±1,9%, no primeiro trimestre, para 25,5±20,3%, no segundo. A proporção volumétrica de citoplasma e núcleo de células lúteas diminuiu ao longo da gestação, e a de tecido conjuntivo aumentou. A quantidade de células lúteas maiores contendo grânulos também aumentou ao longo da gestação.(AU)

The purpose of this study was to evaluate, in cycling and pregnant Nelore cows, the volumetric proportion of the corpus luteum and the distribution of lutein cells with cytoplasmic granules. Twenty four ovaries with corpus luteum were collected in a slaughterhouse. The ovaries were distributed into two groups: pregnant and non-pregnant cows. The group of pregnant cows was divided according to the stage of pregnancy, by the crown-rump fetus length: 1st third - until 90 days, 2nd third - between 91 and 180 days and 3rd third - between 181 and 270 days. The corpus luteum were processed for inclusion in paraplast, and slides were stained for analysis in light microscopy. The constituents quantified in the volumetric proportion were: lutein cells cytoplasm and nucleus, connective tissue and fibroblasts, endothelial cells and pericyte and blood vessels. The volumetric proportion of lutein cells cytoplasm decreased significantly from 46.1±2.8% to 37.9±3.5%, and also decreased significantly in the nucleus from 9.2±1.0% to 6.2±1.0% between the first and the second trimester of pregnancy. The amount of connective tissue and fibroblasts increased significantly throughout gestation, with 20.9±5.4% on the first trimester and 34.0±4.2% on the second. Granules were evidenced by the histochemical technique of Xylidine Ponceau and Bromophenol blue, indicating their probable protein composition. The amount of lutein cells with granules increased throughout gestation, but significant difference was observed only between the first (2.3±1.9%) and second trimester (25.5±20.3%). The cytoplasmic and nucleus volumetric proportion decreased during pregnancy and the volumetric proportion of connective tissue increased. The amount of lutein cells with granules increased throughout pregnancy.(AU)

Cryoprotection effectiveness of low concentrations of natural and lyophilized LDL (low density lipoproteins) on canine spermatozoa / Eficiência de baixas concentrações de lipoproteínas de baixa densidade natural e liofilizada na crioproteto de espermatozoides caninos

The aim of this study was to evaluate the use of low concentrations of natural and lyophilized low density lipoprotein (LDL) from hen's egg yolk for cryopreservation of canine semen. Different ammonium sulphate concentrations were tested to extract LDL from egg yolk. The yolk was centrifuged, and LDL was isolated using 10, 20, 40, 45, or 50 percent ammonium sulphate solution (ASS). The LDL-rich floating fraction was collected for chemical characterization. Dry matter content was lowest (P<0.05) in the LDL extracted with the 50 percent ASS. The purification of LDL increased in association with increasing ammonium sulphate concentrations. SDS-PAGE showed that the 50 percent ASS solution yielded a purer fraction of LDL from egg yolk. For semen cryopreservation, TRIS extender was used replacing 20 percent egg yolk (control) by natural or lyophilized LDL using 1, 2, and 3 percent (w/v). Semen was centrifuged (755Xg for 7 min), diluted with one of the extenders, packed into 0.5mL straws (100x106 sperm/mL), and placed in a programmable cryopreservation machine. Thawed semen (37°C/ 30s) was analyzed for sperm motility, morphology, and by the hypoosmotic and epifluorescence tests (CFDA/ PI). Natural LDL extracted with 50 percent ASS was as effective as whole egg yolk to preserve canine frozen sperm when using low concentrations. The lyophilized LDL, mainly in the two higher concentrations tested (2 and 3 percent), was unsuitable to maintain the effectiveness of the LDL cryoprotective effect on dog sperm...

O objetivo deste estudo foi avaliar o uso de baixas concentrações da lipoproteína de baixa densidade (LBD) extraída da gema do ovo de galinha, nas formas natural e liofilizada, na criopreservado do sêmen canino. Diferentes concentrações de sulfato de amônio também foram testadas na extrato da LBD da gema do ovo. A gema foi centrifugada, sendo a LBD isolada usando-se soluto saturada de sulfato de amônio (SSA) nas concentrações de 10, 20, 40, 45 e 50 por cento. A frado rica em LBD foi coletada para caracterizado química. O conteœdo de matéria seca foi menor (P<0,05) na LBD extraída com SSA 50 por cento. A pureza da LBD melhorou medida que se aumentou a concentrado de SSA utilizada. SDS-PAGE mostrou que a SSA 50 por cento produziu uma frado mais pura de LBD oriunda da gema do ovo. Para o congelamento de sêmen, o meio diluidor TRIS teve a gema do ovo a 20 por cento (controle) substituída pela LBD a 1, 2 e 3 por cento (p/v), nas formas natural e liofilizada. O sêmen foi centrifugado (755xg por 7min), diluído em um dos meios diluidores em teste e envasado em palhetas de 0,5mL (100x106 sptz/mL), sendo congelado em máquina de congelamento programável. O sêmen descongelado (37°C/30s) foi analisado quanto motilidade e morfologia espermática e nos testes hiposm-tico e de epifluorescência (DACF/IP). A LBD natural extraída com SSA 50 por cento foi tão eficiente quanto a gema do ovo na preservado do espermatozoide canino congelado nas baixas concentrações testadas. A LBD liofilizada, principalmente as duas maiores concentrações (2 e 3 por cento), não foi adequada para manter o efeito crioprotetor da LBD sobre o espermatozoide canino...

Qualidade seminal e morfometria testicular de coelhos (Oryctolagus cuniculus) suplementados com geleia real / Seminal quality and testicular morphometry of rabbits (Oryctolagus cuniculus) supplemented with royal jelly

Objetivou-se avaliar o efeito da geleia real na qualidade seminal e na morfometria testicular de coelhos. Quatorze coelhos adultos da raça Nova Zelândia foram distribuídos em três grupos: com administração diária de 1mL de água, via oral (SG); administração diária de 0,5mg (0,5G) de geleia real diluída em 1mL de água, via oral; e administração diária de 1,0mL (1,0G) de geleia real diluída em 1mL de água, via oral. O fornecimento de geleia real foi iniciado 30 dias antes das coletas de sêmen, permanecendo durante todo o período de coleta, totalizando 90 dias. Utilizou-se o método da vagina artificial para coleta de sêmen. Foram avaliados os parâmetros físicos e morfológicos do sêmen e os parâmetros de morfometria testicular. Houve diferença no volume seminal do 0,5G (0,54±0,22) em relação ao SG (0,39±0,13) e ao 1,0G (0,30±0,09) (P<0,05). Para os grupos SG, 0,5G e 1,0G, não houve diferença (P>0,05) para turbilhonamento espermático, concentração espermática, motilidade progressiva e vigor espermático. Os defeitos maiores no grupo 0,5G (8,52±3,26) foram menores do que nos grupos SG (14,09±4,26) e 1,0G (16,1±3,95) (P<0,05). Não houve diferença entre os defeitos menores e os defeitos totais (P>0,05). Os pesos corporal, testicular, epididimário e o índice gonadossomático não diferiram entre os grupos (P>0,05). A ingestão diária de 0,5mg de geleia real apresentou efeitos positivos na morfologia espermática de coelhos.

A trial was carried out to evaluate the effects of royal jelly on the seminal quality and testicular morphometry of rabbits. Fourteen mature rabbits of New Zealand breed were distributed between three groups. The first group was supplied with 1mL of water only (SG), the second group was supplied with 0.5mg of royal jelly diluted in 1mL of water (0.5G), and the third group was supplied with 1mg of royal jelly also diluted in 1mL of water (1.0G). The royal jelly supply started 30 days before semen collection and lasted the entire experimental period. An artificial vagina was used to collect the rabbits' semen. Physical and morphological parameters in the semen and the testicular morphometry were evaluated. Differences were found on the seminal volume in group 0.5G(0,54±0,22) in relation to SG (0,39±0,13) and 1.0G (0,30±0,09) groups (P<0.05). For SG, 0.5G and 1.0G groups, no differences (P>0.05) were found in sperm concentration, gross motility, individual motility and vigor. The total of primary defects in group 0.5G (8,52±3,26) was lower than in groups SG (14,09±4,2) and 1.0G (16,1±3,95) (P<0.05). No significant difference was found between secondary defects and the total defects on the semen (P>0.05). Body, testicular and epididymal weights did not differ between groups, as well to the gonadosomatic index (P>0.05). The ingestion of royal jelly produced positive results on the seminal production of males.

Microscopia e morfometria de componentes tubulares de testículos de coelhos suplementados com geleia real / Microscopy and morphometry of tubular components of rabbit testis supplemented with royal jelly

Estudaram-se os efeitos da geleia real sobre a espermatogênese de coelhos tratados com diferentes concentrações de geleia real. Os tratamentos foram formados por três grupos: grupo-controle; grupo que recebeu 0,5mg/dia de geleia real; e grupo que recebeu 1,0mg/dia de geleia real. O estudo envolveu a morfometria testicular. Não houve diferença entre os tratamentos quanto aos pesos corporal (T1=3,20±0,19kg, T2=2,96±0,30kg e T3=3,21±0,37kg) e gonadal (T1=2,36±0,33g, T2=2,53±0,33g e T3=2,64±0,39g) e quanto aos índices gonadossomático (T1=0,15±0,02%, T2=0,17±0,03% e T3=0,16±0,02%) e tubulossomático (T1=0,06±0,01%; T2=0,07±0,01% e T3=0,06±0,01%). O diâmetro médio dos túbulos seminíferos (T1=225,95±13,27µm, T2=239,68±21,50µm e T3=231,57±15,94µm), a altura do epitélio seminífero (T1=66,05±5,37µm, T2= 73,47±9,11µm e T3=63,34±4,79µm) e o comprimento de túbulos seminíferos por testículo (T1=46,63±13,44m, T2=43,58±12,17m e T3=46,96±9,54m) e por grama de testículo (T1=19,50±2,68m, T2=17,12±3,91m e T3=17,78±1,98m) não diferiram entre tratamentos. Conclui-se que a suplementação com geleia real, nas doses utilizadas, não altera as características testiculares avaliadas.

This study aimed to investigate the effects of royal jelly on spermatogenesis in rabbits treated with different concentrations of RJ (Control; 0,5mg/day; and 1,0mg/day) using testicular morphometry. There was no significant difference between the body weight (T1= 3.20±0.19kg; T2= 2.96±0.30kg; T3=3.21±0.37kg) and gonadal weight (T1= 2.36±0.33g; T2= 2.53±0.33g; T3= 2.64±0.39g), gonadossomatic index (T1= 0.15±0.02%; T2= 0.17±0.03%. T3= 0.16±0.02%) and tubulossomatic index (T1= 0.06±0.01%; T2= 0.07±0.01%. T3= 0.06±0.01%) between treatments, showing that the percentage of body mass, and the percentage of seminiferous tubules allocated in testis were similar in the 3 experimental groups. Similarly, the mean diameter of the seminiferous tubules (T1= 225.95±13.27µm; T2=239.68±21.50µm; T3= 231.57±15,94µm), the height of the seminiferous epithelium (T1=66,05±5,37µm; T2=73.47±9.11µm; T3=63.34±4.79 µm) and length of seminiferous tubule for testis (T1=46.63±13.44m; T2=43.58±12.17m; T3=46.96±9.54m) and per gram of testis (T1=19.50±2.68m; T2=17.12±3.91m; T3=17.78±1.98m) did not differ statistically. It was concluded that supplementation with royal jelly, at the doses used, did not alter the testicular parameters evaluated here.

Parâmetros morfofisiológicos testiculares de camundongos (Mus musculus) suplementados com geleia real / Morphophysiological parameters of mice (Mus musculus) testicles supplemented with royal jelly

Avaliaram-se os efeitos da geleia real sobre os parâmetros morfofisiológicos testiculares de camundongos (Mus musculus). Utilizaram-se 57 machos Swiss, com quatro meses de idade, distribuídos aleatoriamente em seis tratamentos: T1: solução fisiológica, via intraperitoneal; T2: 0,1mg de geleia real, via intraperitoneal; T3: 0,2mg de geleia real, via intraperitoneal; T4: água destilada, via oral; T5: 0,1mg de geleia real, via oral; e T6: 0,2mg de geleia real, via oral. Após 45 dias de suplementação com geleia real, os animais sacrificados e pesados tiveram seus testículos coletados, incluídos em parafina e corados com hematoxilina/eosina. Não houve diferença entre os tratamentos quanto aos: pesos corporal e testicular, índice gonadossomático, diâmetro tubular, altura do epitélio, comprimento total dos túbulos seminíferos, comprimento tubular por grama de testículo, índices tubulossomático e leydigossomático e valores de proporção volumétrica referentes à túnica própria, epitélio seminífero, vaso sanguíneo e vaso linfático. Foi encontrada diferença entre T1 e T3 em relação aos túbulos seminíferos e ao espaço intertubular.

The effects of royal jelly on the morphophysiological parameters of mice (Mus musculus) testicles were studied. Fifty-eight male Swiss mice were evaluated. They were four-month old and were randomly distributed in six treatments: T1: physiological solution, intraperitonial route; T2: 0.1mg of royal jelly, intraperitonial route; T3: 0.2mg of royal jelly, intraperitonial route; T4: distilled water, orally; T5: 0.1mg of royal jelly, orally; and T6: 0.2mg of royal jelly, orally. After 45 days of supplementation with royal jelly, the animals were weighted, slaughtered, and the testicles collected, included in paraffin, and stained with haematoxylin-eosin. No differences among treatments were observed for: body and testicular weights, gonadossomatic index, tubular diameter, epithelial height, total length of seminiferous tubules, tubular length per testicle gram, tubulossomatic and leydigossomatic indexes and the value of volumetric proportion related to tunic, seminiferous epithelium, blood vessel, and lymphatic vessel. Differences between T1 and T3 about the seminiferous tubules and intertubular space were found.

Preservação do sistema genital de chinchila (Chinchilla lanigera) para avaliação em microscopia de luz / Preservation of chinchilla genital system (Chinchilla lanigera) for microscopic evaluation

The preservation of chinchilla genital organs using a fixed solution of paraformaldehyde 10 percent buffered and a saturated solution of Bouin during 4, 12 and 18 hours of fixation, inclusion in paraffin and staining with hematoxylin-eosin was evaluated. The Bouin solution with 12 hours of fixation was the best protocol of fixation for the ovaries, oviduct, uterus and vagina, resulting in little tissue retraction and better cellular integration when compared to the other fixing times (4 and 18 hours). The fixation with paraformaldehyde showed good results of tissues fixation. Chinchilla genital organs can be preserved with paraformaldehyde 10 percent buffered and Bouin solution using 12 hours of fixation.

Proporção volumétrica dos constituintes do corpo lúteo de Nelore / Volumetric proportions of the corpus luteum constituints of Nelore

Volumetric proportions and nuclear diameter of the small and large luteinic cells of the corpus luteum (CL) were evaluated in ovaries of 17 Nelore cows and heifers, collected in slaughterhouse and classified into two groups: group I (GI, n=7), non-pregnant animals, and group II (GII, n=10), pregnant animals. The CL was reduced to small cuts (less than 3mm), which were fixed in Bouin solution and processed for morphometric analysis. The volumetric proportion analysis showed higher mean in the GI animals for the nuclei and cytoplasm of luteinic cells, while the mean of connective tissue and fibroblasts was higher in the GII animals, while the mean of the capillary endothelial cells and pericytes did not differ between the groups. The average nuclear diameter of the large and small luteinic cells did not differ between the groups.

Características de ovários de fêmeas zebu (Bos taurus indicus), colhidos em abatedouros / Characteristics of the ovaries of zebu cows (Bos taurus indicus) collected in slaughterhouses

The ovarian characteristics of pregnant and non pregnant zebu cows (Bos taurus indicus) were studied. The ovaries were obtained in slaughterhouses, identified and divided into: group I, non pregnant animals (GI, n=43) and group II, pregnant animals (GII, n=43). The ovaries were measured and cut in their lenght for measuring and evaluation of the corpus luteum (CL) characteristics. For the GI the measures for the left ovary were 2.5 ± 0.70, 1.61 ± 0.32 and 1.22 ± 0.39cm, respectively for length, width and thickness, and 2.62 ± 0.54, 1.71 ± 0.36 and 1.21 ± 0.31cm for the right ovary. For the GII the measures for the left ovary were 2.78 ± 0.48, 1.80 ± 0.33 and 1.23 ± 0.35cm, respectively for length, width and thickness and 2.84 ± 0.50, 1.74 ± 0.45 and 1.21 ± 0.40cm for the right one. Both groups presented 51.2 percent of CL in the left ovary and 48.8 percent in the right one, with high incidence of CL enclosed and without cavity. The mean for the diameter of the CL was 1.59±0.32cm for GI e 1.82±0.35cm for GII. The results suggest the need to consider carefully the size of the ovary and the enclosed characteristics of the CL when performing gynecological examination of zebu cows via rectal palpation

Preservaçäo do sistema genital de cabras para a avaliaçäo em microscopia de luz / Preservation of goat genital system for microscopic evaluation purpose

The preservation of the goat genital organs with a fixed solution of paraformaldehyde 10 per cent and a saturated solution of Bouin during 4, 8, 12 and 24 hours of fixation, included in paraffin and stained with hematoxylin-eosin was evaluated. The Bouin solution with four hours of fixation showed to be the best protocol of fixation for the ovary, oviduct, uterus and vagina, resulting in little tissue retraction and better cellular integration when compared to the other fixing times (8, 12 and 24 hours). The fixation with paraformaldehyde showed significant alteration in the tissue architecture of the oviduct and vagina. Althouh paraformaldehyde is not the most adequate solution for the goat genital system preservation, its use can be considered in the absence of Bouin solution

Hipoinsulinemia em alcoolistas com hepatopatia mínima / Hypoinsulinemia in alcoholics with minimal liver disease

OBJETIVO: Em alcoolistas portadores de lesões hepáticas mínimas avaliar os níveis de glicose e insulina séricas após estímulo com glicose intravenosa. MÉTODOS: Em oito etilistas, portadores de alterações hepáticas mínimas, caracteriza por biópsia hepática, e em 26 controles sadios não-alcoólicos, foram estudados os níveis glicêmicos e insulinêmicos (RIE) nos tempos 1, 3, 5, e 10 minutos após estímulo com glicose intravenosa (0.5g/Kg de peso). RESULTADOS: As médias da insulina sérica dos tempos 1, 3 minutos e resposta integrada total (RIT-10min) após estímulo foram menores no grupo alcoolista em relação ao controle (p < 0,05) apesar de curvas glicêmicas normais. CONCLUSÃO: Etilistas crônicos com lesões hepáticas mínimas, à semelhança do observado anteriormente em etilistas assintomáticos, apresentam níveis diminuídos de insulina sérica, mas com curvas glicêmicas normais, após estímulo com glicose intravenosa, indicando hipofunção de células beta do pâncreas.

Pancreatite hereditária / Hereditary pancreatitis

Pancreatite hereditaria (PH) é uma causa rara de pancreatite crônica cuja manifestaçäo clínica mais comum é a dor abdominal recorrente que se inicia na infância ou na adolescência. OBJETIVO. Relatar um caso de PH com apresentaçäo atipica e revisäo da literatura. MÉTODOS. Estudou-se um paciente näo etilista, sem história de dor abdominal, que apresentava quadro de esteatorréia e desnutriçäo. A investigaçäo diagnóstica revelou a presença de pancreatite crônica avançada. Dois outros casos semelhantes foram detectados na família. Aspectos clínicos e epidemiológicos desta entidade foram revisados. CONCLUSäO. PH, embora incomum, deve fazer parte do diagnóstico diferencial das pancreatites crônicas, efetuando-se a triagem familiar diante da suspeita clínica

Blood glucose and insulin response to intravenous glucose by patients with chronic Chagas' disease and alcoholism

Blood glucose and insulin responses to intravenous glucose 500 mg/kg in a group of alcoholic chagasic patients (Group I, N = 15) were assessed and compared to those of a group of patients with Chagas' disease without alcoholism (Group II, N = 16), a group of alcoholics without Chagas' disease (Group III, N = 22), and a group of non-alcoholics without Chagas' disease (Group IV, N = 15). Determinations of serum glucose and insulin were done under fasting conditions, 5 and 10 min after the injection of 50 per cent glucose. There was no difference in mean serum glucose levels between groups. Mean serum insulin levels at times 5 and 10 min and for the total integrated response were lower (P < 0.05) in Groups I and II as compared to Group IV; at time 10 min the mean was lower (P < 0.05) in Group III as compared to Group IV. There was a nonsignificant trend for a lower insulinemic response in Group I as compared to Group II. These data suggest denervation of or injury to pancreatic B cell by Chagas' disease