Effects of the Chronic Use of Acetaminophen on Pregnant Rats

Purpose: to evaluate the effects of acetaminophen on the pregnancy of female albino rats. Methods: forty pregnant rats were separated into four groups. All the animals received daily by gavage 1 ml of acetaminophen solution from the first day (day zero) until the 20th day of pregnancy: group I - only distilled water (control); groups II, III and IV, respectively, 125, 500 and 1,500 mg/kg body weight of acetaminophen dissolved in distilled water. The animals were weighed on days 0, 7, 15 and 20 of pregnancy. Results: our results showed that the rats that received the medication presented a reduction in weight when compared to the control group. The incidence of reabsorption of the embryos was 2.0, 3.5 and 7.0 times higher than in the control, in groups II, III and IV, respectively. Groups GII and GIV showed a clear reduction in the weight of the concepts. In GIV there was a 50% reduction in weight increase of fetuses and placentas when compared to the control, and 15.7% of external malformations were also found. Conclusions: the continuous use of acetaminophen should be avoided at doses higher than 70 mg/kg per during pregnancy.Objetivos: estudar a ação do acetaminofeno administrado durante toda a prenhez da rata albina. Métodos: 40 ratas albinas prenhes foram distribuídas ao acaso em quatro grupos numericamente iguais: GI - recebeu diariamente 1 ml de água destilada por gavagem (controle); GII, GIII e GIV foram tratadas, respectivamente, com 125, 500 e 1.500 mg/kg de peso corporal de acetaminofeno, dose única, diária, por gavagem, sempre diluído em 1 ml de água destilada, desde o dia zero até o 20º dia de prenhez. O ganho de peso das matrizes foi avaliado nos dias 0, 7, 15 e 20 de prenhez, sendo que no 20º dia todos os animais foram sacrificados. Resultados: nossos achados mostraram sempre menor peso entre as matrizes que receberam o medicamento, quando comparadas com o controle. A incidência de reabsorção dos embriões foi da ordem de 2,0, 3,5 e 7,0 vezes maior que no controle, considerando-se, respectivamente, os grupos GII, GIII e GIV. Os grupos GIII e GIV tiveram redução do peso dos conceptos. No GIV, houve redução de 50% no crescimento de fetos e placentas com relação ao controle; aqui, também foram encontradas 15,7% de malformações externas. Conclusões: deve ser evitado o uso contínuo do acetominofeno em altas doses, superiores a 70 mg/kg por dia, durante o período gestacional.UNISA Faculdade de Medicina de Santo AmaroUniversidade Federal de São Paulo (UNIFESP) Escola Paulista de Medicina Departamento de TocoginecologiaUNIFESP, EPM, Depto. de TocoginecologiaSciEL

Effects of isoflavones on the skin of postmenopausal women: a pilot study

OBJECTIVE: The aim of this study was to evaluate the effects of isoflavones on the skin of postmenopausal women. DESIGN: A prospective study was performed with 30 postmenopausal women before and immediately after the end of treatment with 100 mg/day of an isoflavones-rich, concentrated soy extract for six months. A skin punch was performed in the gluteal region for sample collection before and immediately after the treatment program. Morphometric determination of epidermal thickness, the papillary index (wrinkling), and the amount of dermal elastic and collagen fibers was assessed. In addition, the number of blood vessels in the sample was also evaluated. The paired Student's t-test was used for statistical analysis (P < 0.05). RESULTS: Isoflavone treatment resulted in a 9.46% increase in the thickness of the epidermis in 23 patients. In addition, the papillary index was reduced in 21 women. The papillary index was inversely proportional to skin wrinkling, i.e., there were a large number of papillae after treatment. The amount of collagen in the dermis was increased in 25 women (86.2%). In 22 women (75.8%) we observed that the number of elastic fibers increased. The number of dermal blood vessels was significantly increased in 21 women. CONCLUSION: Our data show that the use of a concentrated, isoflavone-rich soy extract during six consecutive months caused significant increases in epithelial thickness, the number of elastic and collagen fibers, as well as the blood vessels.Federal University of São Paulo Gynecology DepartmentFederal University of São Paulo Morphology DepartmentUniversidade de São Paulo Faculdade de Medicina Laboratório de Ginecologia Estrutural e MolecularUniversidade de São Paulo Faculdade de Medicina Departamento de Obstetrícia e GinecologiaUNIFESP, Gynecology DepartmentUNIFESP, Morphology DepartmentSciEL

Proliferation of the superficial epithelium of ovaries in senile female rats following oral administration of conjugated equine estrogens

OBJECTIVE: To evaluate the effect of different concentrations of estrogen on the ovarian superficial epithelium in senile female rats. Design: Fifty female rats at 15 months of age and with irregular estrous cycles were selected and randomly divided into five experimental groups containing equal numbers of animals in each: GPROP, control group receiving vehicle only; GE0.05mg, group receiving conjugated equine estrogens (CEE) at a dose of 50 µg/kg; GE0.5mg, group receiving CEE at 500 µg/kg; GE1mg, group receiving CEE at 1 mg/kg; and GE2mg, receiving CEE at 2 mg/kg. The length of treatment was 21 days. After this period, the animals were anesthetized and the ovaries were fixed in 10% formaldehyde and processed for routine histology. Histomorphology was analyzed by light microscopy, and histomorphometrics were evaluated using the Imagelab program. RESULTS: In the GPROP and GE0.05mg groups, the superficial epithelium of the ovary had a simple cuboidal shape, and as the estrogen dose increased, the epithelium thickened, with pseudo-stratified or stratified epithelium appearing in the GE2mg group. The animals in the group given the highest estrogen dose (GE2mg) showed the thickest ovarian epithelium and the largest perimeter and surface area of the surface ovarian epithelium (P < 0.01). However, the difference in epithelium thickness between the GE0.5mg and GE1mg groups was only slight. CONCLUSION: Our data suggest that CEE at a dose of 2 mg/kg may induce marked proliferation of rat ovarian epithelium.UNIFESP Morphology Department Histology and Cellular Biology DivisionUniversidade de São Paulo Faculdade de MedicinaUniversidade Federal de São Paulo (UNIFESP) Faculdade de Medicina Gynecology DepartmentUNIFESP, Morphology Department Histology and Cellular Biology DivisionUNIFESP, Faculdade de Medicina Gynecology DepartmentSciEL

Seminal quality, testicle and epididymis morphology of bulls fed a diet containing gossypol

The objective was to evaluate the effect of intake of free gossypol on sperm quality and morphology of the testicles and epididymis of Nelore bulls. Twelve bulls were fed diets containing 3.3g of free gossypol/bull/day (cottonseed) (Group 1, n = 6) and a diet free of gossypol (Group 2, n = 6), respectively. Semen samples were collected in the beginning and end of the experiment which lasted 73 days. In the end of the study the testes and epididymis of bulls were removed to study the effect of free gossypol on histological characteristics. The average consumption of 3.3g of free gossypol/bull/day (mean 7.1mg of free gossypol/kg/day) reduced motility and sperm concentration and increased the percentage of major and total sperm defects, as well as the animals showing testes with seminiferous tubules of smaller thickness, fewer layers of spermatogenic lineage cells, smaller epididymal epithelium thickness and smaller number of sperm within the epididymal ducts, compared to animals with a diet free of gossypol (Group 2). The consumption of 3.3g of free gossypol/bull/day led to changes in morphology and morphometry of the testes and epididymis and reduced sperm quality of bulls.Objetivou-se avaliar o efeito da ingestão de gossipol livre sobre a qualidade espermática e a morfologia dos testículos e dos epidídimos de touros da raça Nelore. Doze touros receberam dieta contendo 3,3g de gossipol livre/touro/dia (caroço de algodão) (Grupo 1, n=6) e dieta isenta de gossipol (Grupo 2, n=6), respectivamente. Foram realizadas coletas de sêmen no início e no final do experimento, que teve duração de 73 dias. Ao final do estudo, foram retirados os testículos e os epidídimos dos touros para se estudar o efeito do gossipol livre sobre as características histológicas. O consumo médio de 3,3g de gossipol livre/touro/dia (média 7,1mg de gossipol livre/kg/dia) reduziu a motilidade e a concentração espermática e aumentou a porcentagem de defeitos espermáticos maiores e totais. Além disso, os animais apresentaram testículos com túbulos seminíferos de menor espessura de parede, menor número de camadas de células espermatogênicas, menor espessura do epitélio epididimário e menor número de espermatozoide no interior dos ductos epidídimários, em relação aos animais com dieta isenta de gossipol (Grupo 2). O consumo de 3,3g de gossipol livre/touro/dia acarretou alterações na morfologia e na morfometria dos testículos e dos epidídimos e reduziu a qualidade espermática dos touros.Universidade de Cuiabá Faculdade de Medicina VeterináriaUFMS Faculdade de Medicina Veterinária e ZootecniaEpamigUniversidade Federal do Mato GrossoUNIFESP-EPMUNIFESP, EPMSciEL

Glycosaminoglycan distribution in the rat uterine cervix during the estrous cycle

OBJECTIVE: To analyze the amount of glycosaminoglycans in the uterine cervix during each phase of the rat estrous cycle. DESIGN: Based on vaginal smears, forty female, regularly cycling rats were divided into four groups (n = 10 for each group): GI - proestrous, GII - estrous, GIII - metaestrous and GIV - diestrous. Animals were sacrificed at each phase of the cycle, and the cervix was immediately removed and submitted to biochemical extraction and determination of sulfated glycosaminoglycans and hyaluronic acid. The results were analyzed by ANOVA followed by the Bonferroni post-hoc test. RESULTS: The uterine cervix had the highest amount of total sulfated glycosaminoglycans and dermatan sulfate during the estrous phase (8.90 ± 0.55 mg/g of cetonic extract, p<0.001; and 8.86 ± 0.57 mg/g of cetonic extract, p<0.001). In addition, there was more heparan sulfate at the cervix during the proestrous phase (0.185 ± 0.03 mg/g of cetonic extract) than during any other phase (p<0.001). There were no significant changes in the concentration of hyaluronic acid in the uterine cervix during the estrous cycle. CONCLUSION: Our data suggest that the amount of total sulfated glycosaminoglycans may be influenced by hormonal fluctuations related to the estrous cycle, with dermatan sulfate and heparan sulfate being the glycosaminoglycans most sensitive to hormonal change.Universidade Federal de São Paulo (UNIFESP) Departmento de Morfologia e GenéticaUniversidade de São Paulo Faculdade de Medicina Departamento de Obstetrícia e GinecologiaUniversidade de São Paulo Instituto de Ciências Biomédicas Department of PharmacologyEscola Paulista de Medicina Department of GynecologyUNIFESP, Departmento de Morfologia e GenéticaEscola Paulista de Medicina Department of GynecologySciEL

Endometrium in women with polycystic ovary syndrome during the window of implantation

The human endometrium undergoes to a complex series of prolifertive and secretory changes in each menstrual cycle and displays only a short period of receptivity, known as the window of implantation, necessary for the implantation of the blastocyst in the uterus. The implantation process occurs in a sequential manner, leading to the establishment of pregnancy. Morphofunctional changes during this period may prevent or hinder the implantation. For this reason, the study of the endometrium at this stage is important for the improvement of therapies that may interfere with the mechanisms involved in maternal-embryonic interaction. Several gynecological disorders, including polycystic ovary syndrome (PCOS), are associated with decreased fertility and uterine receptivity. In spite of recent advances in assisted reproduction techniques, allowing the selection of high quality embryos, the implantation rate remains low and has not increased enough in recent decades. This article aims at reviewing the endometrial aspects of the window of implantation in women with polycystic ovary syndrome, focusing mainly on adhesion molecules. For that purpose, we analyzed 105 articles published in journals indexed in PubMed in the last 50 years (up to May 2011). In conclusion, the endometrial receptivity seems to be the major limiting factor for the establishment of pregnancy in a large number of gynecological diseases, including PCOS, and treatment to improve implantation rates is likely to be taken towards this direction.O endométrio humano é submetido a uma complexa série de mudanças proliferativas e secretórias em cada ciclo menstrual e exibe somente pequeno período de receptividade, conhecido como janela de implantação, necessário para a nidação do blastocisto no útero. O processo da implantação ocorre de forma sequencial, levando ao estabelecimento da gravidez. Alterações morfofuncionais durante este período podem impedir ou dificultar a implantação. Por este motivo, o estudo do endométrio nesta fase é importante para o aprimoramento de terapias que possam interferir nos mecanismos envolvidos na interção materno-embrionária. Várias doenças ginecológicas, incluindo a síndrome dos ovários policísticos (SOP), estão associadas à diminuição da fecundidade e da receptividade uterinas. Apesar de recentes avanços nas técnicas de reprodução assistida, permitindo a seleção de embriões de alta qualidade, a taxa de implantação continua baixa e não tem aumentado suficientemente nas últimas décadas. O presente artigo tem como objetivo revisar os aspectos endometriais da janela de implantação em mulheres com a síndrome dos ovários policísticos, focando especialmente as moléculas de adesão. Para tanto, nos valemos da análise de 105 artigos publicados em revistas indexadas no PUBMED nos últimos 50 anos (até maio de 2011). Como conclusão, a receptividade endometrial parece ser o maior fator limitante no estabelecimento da gestação em grande número de doenças ginecológicas, incluindo a SOP, e o tratamento para melhorar as taxas de implantação possivelmente será nessa direção.Universidade Federal de São Paulo (UNIFESP) Departamento de GinecologiaUniversidade Federal do PiauíUniversidade de São Paulo Faculdade de MedicinaUNIFESP Departamento de MorfologiaFMUSP Departamento de Obstetrícia e GinecologiaUNIFESP, Depto. de GinecologiaUNIFESP, Depto. de MorfologiaSciEL

Does Gestrinone Antagonize the Effects of Estrogen on Endometrial Implants Upon the Peritoneum of Rats?

OBJECTIVE: To evaluate the effects of estrogen treatment in combination with gestrinone on an experimental rat model of endometriosis. METHODS: Uterine transplants were attached to the peritoneum of female Wistar rats via a surgical autotransplantation technique. The implanted area was measured during the proestrus phase and after hormonal treatment. We performed morphometric analysis and examined the macroscopic and morphometric alterations of endometrial implants after hormonal treatment in ovariectomized rats. RESULTS: The high dose of estrogen caused macroscopic increases in the endometrial implant group compared with other groups, which were similar to increases in the proestrus phase. The low dose showed morphometric development of implants, such as an increase in number of endometrial glands, leukocyte infiltration and mitosis. Gestrinone antagonized both doses of estrogen. CONCLUSION: Our findings suggest that gestrinone antagonizes estrogen's effects on rat peritoneal endometrial implants

Effects of combined estrogen and raloxifene therapy on rat endometrium

PURPOSE: to evaluate the effects of conjugated equine estrogens (CEE) and raloxifene (Ral), alone or combined, on the rat endometrium. METHODS: fifty-six adult rats were ovariectomized and randomly divided into seven groups: GCont (control); GCEE (CEE 50 µg/kg); GCEE/25 (CEE 25 µg/kg); GRal/0.75 (Ral 0.75 mg/kg); GRal/0.4 (Ral 0.4 mg/kg); GCEERal (50/0.75) - (CEE 50 µg/kg + Ral 0.75 mg/kg), and GCEE-Ral (25/0.4) - (CEE 25 µg/kg + Ral 0.4 mg/kg). The drugs were orally administered (gavage) for 21 consecutive days. At the end of the experiment, all animals were anesthetized and sacrificed. Fragments of uterus were removed, fixed in 10% formaldehyde and processed for paraffin inclusion. The histological sections were stained by HE and submitted to histomorphometric evaluation. The following parameters were analyzed: thickness of superficial epithelium and number of endometrial glands/mm² and of blood vessels/mm². The data were evaluated using ANOVA followed by the Turkey-Kramer test. RESULTS: in the GCont and only Ral treatment (GRal/0.75 and GRal/0.4) the endometrium showed signals of atrophy. In the groups treated with only CEE signs of endometrial proliferation were observed, mainly in group GCEE/50. Also, there was endometrial proliferation in the groups that received combined CEE and Ral (Ral GCEE (50/0.75) and GCEE-Ral (25/0.4)), but it was more intensive in the animals treated with isolated estrogen than in those that received combined estrogen and raloxifene. CONCLUSION: raloxifene may partially block the action of estrogen on the castrated adult rat endometrium.OBJETIVO: avaliar os efeitos dos estrogênios conjugados eqüinos (Ece) e do raloxifeno (Ral), isoladamente ou associados, sobre o endométrio de ratas adultas. MÉTODOS: foram utilizadas 56 ratas adultas, ooforectomizadas, divididas aleatoriamente em sete grupos: GCont (controle); GEce (Ece 50 µg/kg); GEce/25 (Ece 25 µg/kg); GRal/0,75 (Ral 0,75 µg/kg); GRal/0,4 (Ral 0,4 mg/kg); GEce-Ral (50/0,75) - (Ece 50 µg/kg + Ral 0,75 mg/kg) e GEce-Ral (25/0,4) - (Ece 25 µg/kg + Ral 0,4 mg/kg). As drogas foram administradas por gavagem durante 21 dias consecutivos. Ao final da administração todos os animais foram anestesiados e fragmentos dos úteros removidos, fixados em formol a 10% e processados para inclusão em parafina. Os cortes obtidos foram corados por HE e submetidos à avaliação histomorfométrica. Foram avaliados os seguintes parâmetros: espessura do epitélio superficial, número de glândulas endometriais/mm² e número de vasos sangüíneos presentes no miométrio/mm². Os dados obtidos foram submetidos a análise estatística de ANOVA seguida pelo teste de Tukey-Kramer. RESULTADOS: nos grupos controle (Gcont) e tratados isoladamente com raloxifeno (GRal/0,75 e GRal/0,4) o endométrio mostrou-se atrófico. Já nos grupos tratados com Ece isoladamente foi observado endométrio muito desenvolvido, sendo este efeito mais acentuado no grupo que recebeu 50 µg/kg, no qual encontramos aumento da espessura do epitélio superficial e da lâmina própria e no número de glândulas endometriais e de vasos sanguíneos. Notamos também proliferação endometrial nos grupos que receberam a associação de Ece e Ral (GEce-Ral - 50/0,75) e (GEce-Ral - 25/0,4). CONCLUSÃO: o raloxifeno parece bloquear parcialmente a ação do estrogênio no endométrio de ratas adultas castradas.Universidade Federal de São Paulo (UNIFESP) Departamento de Ginecologia e ObstetríciaUniversidade Federal de São Paulo (UNIFESP)Universidade Federal de São Paulo (UNIFESP) Departamento de MorfologiaUniversidade Federal de São Paulo (UNIFESP) Departamento de GinecologiaUNIFESP, Depto. de Ginecologia e ObstetríciaUNIFESP, Depto. de MorfologiaUNIFESP, Depto. de GinecologiaSciEL

Morphological and Morphometrical Study of Permanent Estrus Rat Mammary Gland Treated with Danazol

Purpose: the morphologic and morphometric aspects of the breasts of rats in permanent estrus submitted to danazol were studied. Methods: the animals were divided into three groups: group A (n = 12) received water and was used as control, group B (n = 13) was exposed to 20 mg danazol kg-1 day-1, and group C (n = 10) was exposed to 80 mg danazole kg-1 day-1 for 35 consecutive days. The microscopic study evaluated the ductal and acinar distribution. Histometry of the relationship duct/stroma was based on the principles of stereology with a Zeiss k-10X ocular, with Integrationsplatte I reticulum of Weibel of 25 hits, with 100X magnification. For each studied section, 10 aleatoric fields were counted, with a total of 250 points. The variance analysis test (Kruskal-Wallis) was applied to compare the three groups in relation to the mean number of alveoli and ducts (alpha = 0.05). Results: when submitted to morphological study, all groups presented lobules with alveoli lined with cubic cells with nuclei in their central or basal portion. Small amounts of eosinophilic material were observed in some cases in the lumen, with no differences between the groups. At morphometry, with a magnification of 100X, a mean number of 28.6 ducts/10 fields was found in group A, 28.4 in group B and 29.2 in group C (Kruskal-Wallis test: Hcrit = 0.1). The mean number of alveoli in 10 fields was 5.9, 9.3 and 6.5 in groups A, B and C, respectively (Kruskal-Wallis test: Hcrit = 2.9), with no significant differences between the groups. Conclusion: danazol did not cause any changes in the morphology and morphometry of the permanent estrus mammary epithelium.Objetivo: estudar os efeitos do danazol sobre a mama da rata em estro permanente. Métodos: os animais foram divididos em três grupos: os do grupo A (n = 12) receberam água (placebo), como grupo controle; os do grupo B (n = 13) foram expostos a 20 mg/kg de danazol por dia; os do grupo C (n = 10) foram expostos a 80 mg/kg de danazol por dia. A droga foi administrada durante 35 dias consecutivos. O estudo microscópico dos cortes avaliou a distribuição ductal e acinar. A histometria da relação ducto/estroma foi feita baseada nos princípios da estereologia, com ocular k-10X da Zeiss, com retículo de integração (Integrationsplatte I-retículo de Weibel de 25 hits), com aumento final de 100 vezes. Para cada lâmina estudada, foram contados 10 campos aleatórios, num total de 250 pontos. O teste aplicado foi a análise de variância por postos de Kruskal-Wallis, para comparar os três grupos em relação ao número médio de alvéolos e ductos nas mamas (alfa = 0,05). Resultados: em todos os grupos, os lóbulos apresentaram ao estudo morfológico alvéolos revestidos com células cúbicas com núcleos na sua porção central ou basal. Pequenas quantidades de material eosinofílico foram observadas em alguns casos na sua luz. À morfometria (magnificação de 100X) encontraram-se em média 28,6 ductos/10 campos no grupo A, 28,4 no grupo B e 29,2 no grupo C (análise de Kruskal-Wallis: Hcrit = 0,1). As médias dos números de alvéolos /10 campos foram 5,9 (grupo A), 9,3 (grupo B) e 6,5 (grupo C) (Kruskal-Wallis Hcrit = 2.9), sem diferença significativa entre os grupos. Conclusão: o danazol não causou nenhuma alteração significante na morfologia e morfometria do epitélio das mamas das ratas em estro permanente.Universidade Federal de São Paulo (UNIFESP) EPMUNIFESP, EPMSciEL