Abstract Introduction The CA1 region of the hippocampus has an important role in learning and memory. It has been shown that estrogen deficiency may reduce the synaptic density in the region and that hormone replacement therapy may attenuate the reduction. Objectives This study aimed to evaluate the effects of estrogen and raloxifene on the synaptic density profile in the CA1 region of the hippocampus in ovariectomized rats. Methods Sixty ovariectomized three-month-old virgin rats were randomized into six groups (n = 10). Treatments started either three days (early treatment) or sixty days (late treatment) after ovariectomy. The groups received propylene glycol vehicle (0.5 mL/animal/day), equine conjugated estrogens (50 μg/animal/day), or raloxifene (3 mg/kg/day) either early or late after ovariectomy. The drugs were administered orally by gavage for 30 days. At the end of the treatments, the animals were anesthetized and transcardially perfused with ether and saline solution. The brains were removed and prepared for analysis under transmission electron microscopy and later fixed. Results Results showed a significant increase in the synaptic density profile of the hippocampal CA1 region in both the early estrogen (0.534 ± 0.026 µ/m2) and the early raloxifene (0.437 ± 0.012 µ/m2) treatment groups compared to the early or late vehicle-treated control groups (0.338 ± 0.038 µ/m2 and 0.277 ± 0.015 µ/m2 respectively). Conclusions The present data suggest that the raloxifene effect may be lower than that of estrogen, even early or late treatment, on synaptic density in the hippocampus. CA memory reduction threemonthold month old n 10. 10 . 10) ovariectomy 0.5 05 0 5 (0. mL/animal/day, mLanimalday mL/animal/day , mL animal day mL/animal/day) 50 (5 μg/animal/day, μganimalday μg/animal/day μg μg/animal/day) 3 ( mg/kg/day mgkgday mg kg treatments solution fixed 0.534 0534 534 (0.53 0026 026 0.02 µm2 µm µ m2 m 0.437 0437 437 (0.43 0012 012 0.01 vehicletreated treated 0.338 0338 338 (0.33 0038 038 0.03 µ/m 0277 277 0.27 0015 015 respectively. respectively respectively) 1 0. (0 0.53 053 53 002 02 0.0 0.43 043 43 (0.4 001 01 0.33 033 33 (0.3 003 03 027 27 0.2 00 0.4 04 4 0.3 2

Abstract Purpose: Gelfoam scaffold is a feasible and safe non-invasive technique for Adipose tissue-derived Stem Cell (ASC)-delivery in the treatment of frozen-thawed ovarian autografts. This study seeks to analyze the genes expression profile of rat frozen-thawed ovarian autografts treated with scaffold-based delivery of adipose tissue-derived stem cells. Methods: Eighteen adult Wistar rats were distributed into three groups: Control (frozen-thawed only); Group 1 (Gl) and Group 2 (G2) (frozen-thawed ovaries treated with culture medium or ASC, respectively). Both treatments were performed immediately after autologous retroperitoneal transplant with scaffold-based delivery. The ovarian grafts were retrieved 30 days after transplantation. Quantitative gene expression (qPCR) for apoptosis, angiogenesis, and inflammatory cytokines (84 genes in each pathway) were evaluated by RT-PCR. Graft morphology (HE), apoptosis (cleaved-caspase-3), neoangiogenesis (VEGF), and cellular proliferation (Ki-67) were assessed. Results: In grafts treated with ASC, the apoptosis pathway showed the highest number of genes over-regulated — 49 genes — compared to inflammation cytokines and angiogenesis pathway — 36 and 23 genes respectively, compared to grafts treated with culture medium. Serpinb5 family was highlighted in the angiogenesis pathway and Cxcl6 in the inflammation cytokines pathway. In the apoptosis pathway, the most over-regulated gene was Cap-sasel4. ASC treatment promoted the reduction of cleaved caspase-3 in the theca internal layer and increased cell proliferation by Ki-67 in the granulosa layer without altering VEGF. A mild inflammatory infiltrate was observed in both groups. Conclusion: ASC therapy in rat frozen-thawed ovarian autografts promoted an abundance of genes involved with apoptosis and inflammatory cytokines without compromising the ovary graft morphology and viability for short time. Further studies are necessary to evaluate the repercussion of apoptosis and inflammation on the graft in the long term. HIGHLIGHTS The scaffold-based delivery therapy with adipose tissue-derived stem cells in the rat ovarian autografts seems to be the best option when compared to direct injection or systemic route. Ovarian grafts treated with adipose tissue-derived stem cells showed the highest number of genes over-regulated in the apoptosis pathway, compared to inflammation cytokines and angiogenesis pathway. Capsase14 was the most over-regulated gene in the apoptosis pathway. The treatment with adipose tissue-derived stem cells in ovarian grafts treated didn’t compromise the ovary graft morphology and viability for short time.

Apresenta-se uma proposta pedagógica, não habitual nas escolas médicas tradicionais, que introduz precocemente o aluno de graduação em um estágio de observação da prática médica. Alunos do segundo semestre do curso médico preparados para fazer uma observação participativa do atendimento aos pacientes e entrevistas com profissionais de saúde, desenvolveram a aprendizagem e a reflexão sobre comunicação do profissional de saúde com sua clientela a partir dos fatos observados. Resultados indicam que a experiência com a disciplina propiciou aos alunos um estilo próprio de entrevistar e abordar o paciente, que enfatiza a integridade do ser e as representações do seu mundo, em contraposição à comunicação médico-paciente que acontece durante a anamnese estruturada e na consulta médica feita na rotina ambulatorial com grande demanda.

We present a pedagogical proposal, which is not common in traditional medical schools. This proposal introduces the graduate student early on to a medical practice observation stage. Second semester students of the medical course were trained to observe patient consultations and interviews with health professionals in a participative way. They developed their learning ability and learned how to reflect on the health professional-client communication process from what they observed. Results indicate hat this experience provided students with their own personal style of approaching and interviewing the patients that emphasizes the integrity of the individual and the way in which he or she looks at their world, as opposed to the doctor-patient communication that happens during a structured anamnesis and in a medical consultation provided in an extremely busy outpatients department.

Se presenta una propuesta pedagógica que no es habitual en las Escuelas de Medicina tradicionales, ella introduce precozmente al alumno de graduación en la observación de la practica medica. Los alumnos del segundo semestre de la Facultad de Medicina se prepararon para hacer una observación participativa de la atención al enfermo y de entrevistas con profesionales de la salud, desarrollaron un conocimiento y una reflexión sobre la comunicación del profesional de la salud con sus pacientes, a través de la acción observada. Los datos apuntan que la experiencia con la disciplina, dio a los alumnos un estilo propio de entrevistas y abordajes a los pacientes que enfatiza la integridad del ser y las representaciones de su mundo, en contraposición a la comunicación médico/enfermo que ocurre durante la entrevista estructurada en la consulta medica que se hace en la rutina corrida del ambulatorio donde hay mucha demanda.

Apresenta-se uma proposta pedagógica, não habitual nas escolas médicas tradicionais, que introduz precocemente o aluno de graduação em um estágio de observação da prática médica. Alunos do segundo semestre do curso médico preparados para fazer uma observação participativa do atendimento aos pacientes e entrevistas com profissionais de saúde, desenvolveram a aprendizagem e a reflexão sobre comunicação do profissional de saúde com sua clientela a partir dos fatos observados. Resultados indicam que a experiência com a disciplina propiciou aos alunos um estilo próprio de entrevistar e abordar o paciente, que enfatiza a integridade do ser e as representações do seu mundo, em contraposição à comunicação médico-paciente que acontece durante a anamnese estruturada e na consulta médica feita na rotina ambulatorial com grande demanda.

We present a pedagogical proposal, which is not common in traditional medical schools. This proposal introduces the graduate student early on to a medical practice observation stage. Second semester students of the medical course were trained to observe patient consultations and interviews with health professionals in a participative way. They developed their learning ability and learned how to reflect on the health professional-client communication process from what they observed. Results indicate hat this experience provided students with their own personal style of approaching and interviewing the patients that emphasizes the integrity of the individual and the way in which he or she looks at their world, as opposed to the doctor-patient communication that happens during a structured anamnesis and in a medical consultation provided in an extremely busy outpatients department.

Se presenta una propuesta pedagógica que no es habitual en las Escuelas de Medicina tradicionales, ella introduce precozmente al alumno de graduación en la observación de la practica medica. Los alumnos del segundo semestre de la Facultad de Medicina se prepararon para hacer una observación participativa de la atención al enfermo y de entrevistas con profesionales de la salud, desarrollaron un conocimiento y una reflexión sobre la comunicación del profesional de la salud con sus pacientes, a través de la acción observada. Los datos apuntan que la experiencia con la disciplina, dio a los alumnos un estilo propio de entrevistas y abordajes a los pacientes que enfatiza la integridad del ser y las representaciones de su mundo, en contraposición a la comunicación médico/enfermo que ocurre durante la entrevista estructurada en la consulta medica que se hace en la rutina corrida del ambulatorio donde hay mucha demanda.