Amanda Maria Sena Reis

Primary Ovarian Rhabdomyosarcoma in a Dog

A 10-year-old female English pointer was diagnosed with an ovarian tumour with abdominal metastases. Ultrasonography revealed several nodules of 1-5 cm diameter within the abdominal cavity. Fine needle aspiration cytology of the nodules suggested a malignant mesenchymal tumour. On necropsy examination the right ovary and its capsule were enlarged and there were white-red, friable nodular masses distributed over the surface of the pancreas, liver, omentum, mesentery and serosae of the small and large intestines. Microscopical evaluation revealed neoplastic cells with a high degree of pleomorphism and vascular invasion. Immunohistochemically, the neoplastic cells expressed myosin, desmin, vimentin and CD10, but were negative for cytokeratin, placental alkaline phosphatase, inhibin-alpha and smooth muscle actin. Based on these findings a diagnosis of primary ovarian alveolar rhabdomyosarcoma was made.

Nonlinear effects of caffeine on the viability, synthesis and gene expression of chondrocytes from the offspring of rats treated during pregnancy

OBJECTIVE Evaluate the effects of doses of caffeine administered to pregnant rats on the articular cartilage chondrocytes of their offspring. METHODS Twenty-four adult Wistar rats were randomly assigned to four groups, with one control group and three groups being treated with caffeine at doses of 25, 50 and 100 mg/kg throughout pregnancy. At birth, three offspring/females were euthanized so that the chondrocytes could be extracted. At 7, 14 and 21 days of culture, the chondrocytes were subjected to the MTT cell viability assay and an evaluation of their alkaline phosphatase activity and collagen synthesis. Chondrocytes were also stained by Hematoxylin-eosin, PAS, Safranin-O and Alcian Blue. The Sox-9, Runx-2, aggrecan, collagen-II and alkaline phosphatase gene transcript levels were also evaluated. Mean comparisons were performed by the Student-Newman-Keuls test. RESULTS Chondrocyte cultures from the 25 mg/kg group had the lowest results, as chondrocytes from this group had reduced viability, percentage of cells, alkaline phosphatase activity and collagen and chondrogenic matrix synthesis. A reduced expression of Sox-9, alkaline phosphatase and collagen-II was also detected in the 25 mg/kg group. Chondrocyte cultures of the group treated with 50 mg/kg caffeine showed reduced collagen synthesis and Sox-9 expression. The caffeine dose of 100 mg/kg also reduced collagen and Sox-9 and alkaline phosphatase expression. CONCLUSION Caffeine administered to pregnant rats negatively alters the articular cartilage chondrocytes of their offspring, reducing the synthesis of collagen and Sox-9 expression regardless of the dose. This study also concluded that the effects of caffeine are not linear or dose-dependent.

Effects of excess maternal thyroxin on the bones of rat offspring from birth to the post-weaning period

Objective To evaluate, in rat offspring, bone changes induced by excess maternal thyroxin during pregnancy and lactation, and to assess the reversibility of these changes after weaning. Material and methods Twenty Wistar rats were distributed in two groups, hyperthyroid and control, that were treated daily with L-thyroxin (50 mcg/animal) and placebo, respectively. The treatment was initiated seven days before mating and continued throughout pregnancy and lactation. From every female of each of the two groups, two offspring were euthanized after birth, two at 21 days of age (weaning), and two at 42 days of age (21 days after weaning). In newborns, the length of pelvic and thoracic limbs were measured, and in the other animals, the length and width of the femur and humerus were measured. Bones were dissected, decalcified, embedded in paraffin, and analyzed histomorphometrically. Results Excess maternal thyroxin significantly reduced the length of the pelvic limb in neonates. In 21-day-old individuals, excess maternal thyroxine reduced the length and the width of the femur and the humerus. It also increased thickness of the epiphyseal plate and the percentage of trabecular bone tissue. In 42-day-old individuals, there were no significant differences between groups in relation to the parameters evaluated in the previous periods. Conclusion Excess maternal thyroxine reduced growth in suckling rats both at birth and at weaning, and it also increased the percentage of trabecular bone tissue in 21-day-old animals. These changes, however, were reversible at 42 days, i.e., 21 days after weaning. Arch Endocrinol Metab. 2016;60(2):130-7.

Efeitos da ingestão de cafeína durante a gestação e a lactação sobre a pele de ratas e de filhotes e sua relação com as concentrações séricas do cortisol materno

O objetivo deste trabalho foi verificar os efeitos da ingestao materna de diferentes doses de cafeina durante a gestacao e a lactacao, na pele de ratas-maes e filhotes, bem como sua relacao com as concentracoes sericas do cortisol materno. Vinte e quatro ratas Wistar adultas foram distribuidas em quatro grupos, representados pelo controle e tratados, com cafeina nas doses de 25, 50 e 100mg/kg. Os grupos tratados receberam cafeina por sonda orogastrica durante toda a gestacao e a lactacao. O controle recebeu agua destilada como placebo. Foram avaliados e quantificados os diferentes tipos de foliculos pilosos e a espessura da epiderme. A tecnica de imuno-histoquimica, com o uso do anticorpo anti-CDC47, foi utilizada para avaliar a proliferacao celular da epiderme e dos foliculos pilosos das maes. Na mae, tambem foram mensurados os niveis sericos de cortisol pela tecnica da quimioluminescencia. Os dados foram submetidos a analise de variância com comparacao das medias pelos testes Kruskal-Wallis e SNK. Nos grupos tratados com cafeina nas doses de 25 e 50mg/kg, tanto as maes quanto seus filhotes apresentaram hipotricose e/ou alopecia focal. Apesar de a frequencia de alteracoes macroscopicas das maes ter sido superior a dos filhotes, nestes as lesoes, quando presentes, foram difusas. A analise histologica demonstrou calcinose de foliculos pilosos nas maes e nos filhotes. Mas a morfometria somente revelou diferenca significativa no numero de foliculos pilosos das maes, bem como reducao significativa da proliferacao celular dos foliculos pilosos do grupo tratado com 50mg/kg de cafeina. Os niveis de cortisol materno somente foram significativamente elevados no grupo tratado com 100mg/kg de cafeina. Conclui-se que a cafeina ingerida pelas ratas gestantes e lactantes pode causar lesoes cutâneas tanto nas maes quanto nos filhotes, caracterizadas por hipotricose e/ou alopecia, independentemente dos niveis sericos do cortisol materno.

In vitro effects of caffeine in growth cartilage of rats

OBJECTIVE: To evaluate the in vitro effects of caffeine on proliferation, apoptosis and gene transcripts expression of chondrogenic differentiation in growth cartilage. METHODS: The cartilaginous epiphyses of femurs of newborn rats, were divided into two subgroups: treated with caffeine and control group, both observed over the time periods of 0, 7, 14 and 21 days. The cartilaginous epiphyses of femurs of each subgroup and each time span were subjected to histomorphometrics, immunohistochemical analysis, tunel technique and RT-PCR in real time. RESULTS: The decrease in proliferative activity and the increase of apoptotic chondroblasts at 21 days were found regardless of the subgroup. However, the decrease in cell proliferation caused by caffeine was lower than in the control group and significantly increased the expression of gene transcripts for chondrogenic differentiation, represented by SOx-9 and RUNx-2. However, the in vitro culture with caffeine revealed antagonistic effects: despite the positive effect on chondroblasts proliferation and differentiation, caffeine increased apoptosis, characterized by increased expression of caspase-3 and of the number of cells undergoing apoptosis (p<0.05). CONCLUSION: Caffeine presents antagonistic effects in vitro on growth cartilage, increasing the proliferation, differentiation and cell apoptosis. Experimental Study.Objective To evaluate the in vitro effetcs of caffeine on proliferation, apoptosis a nd gene transcripts expression of chondrogenic differentiation in growth cartilage. Methods The cartilaginous epiphyses of femurs of newborn rats, which were divided into two subgroups: treated with caffeine and control group, both observed over the time periods of 0, 7, 14 and 21 days. The cartilaginous epiphyses of femurs of each subgroup and each time span were subjected to histomorphometric, immunohistochemical analysis, Tunel technique and RT-PCR in real time. Results The decrease in proliferative activity and the increase of apoptotic chondroblasts at 21 days were found regardless of the subgroup. However, the decrease in cell proliferation caused by caffeine was lower than in the control group and significantly increased the expression of gene transcripts for chondrogenic differentiation, represented by Sox-9 and Runx-2. However, the in vitro culture with caffeine revealed antagonistic effects: despite the positive effect on chondroblasts proliferation and differentiation, caffeine increased apoptosis, characterized by increased expression of caspase 3 and of the number of cells undergoing apoptosis (p<0.05). Conclusion Caffeine presents antagonistic effects in vitro on growth cartilage, increasing the proliferation, differentiation and cell apoptosis. Experimental Study.