Variation in neuronal differentiation of a newly isolated mouse embryonic stem cell line: a detailed immunocytochemistry study.

Neural precursor differentiation from mouse ES (embryonic stem) cells have been demonstrated using EB (embryoid body), co-culture on stromal feeder layers, and in the absence of external inducing signals. Most of available mouse ES cell original research articles have worked with only six different cell lines. Our goals were to isolate one new mouse ES lineage, and perform a detailed immunocytochemistry study during neural differentiation, making use of an EB strategy protocol following the generation of neural progenitors, glial cells and postmitotic neurons. The dynamics of differentiation of ES cell derived neuronal precursors into differentiated glia cells and neurons were followed in vitro and correlated to exposure to specific elements of feeder medium. Morphological aspects of generated cellular types, including its immunocytochemical expression of differentiation markers were studied. Immuno-positivity against ß-III tubulin, PGP and TH (tyrosine hydroxylase) was observed from stage I. Approximately 80% of cells were positive for TH at stage I. The first glial cell type appears in stage III. TH, PGP or ß-III tubulin-positive cells with neuronal typical morphology only being seen in stage III when TH-positive cells corresponded to approximately 12% of total cells. Variations among other literature findings can be explained by the choice we made to use a newly isolated ES cell line. As colonies may behave differently during neuronal differentiation, it reinforces the necessity of studying original ES cell lines.

Dienogest: Uma nova opção terapêutica em endometriose / Dienogest: A new option in the treatment of endometriosis

Endometriose é uma doença crônica que afeta mulheres jovens em idade reprodutiva provocando dor, dispareunia, infertilidade e que também afeta a qualidade de vida das pacientes. Numerosos tratamentos existem, alguns clínicos e outros cirúrgicos. Entretanto, essa doença deve ser considerada como uma doença crônica que pode exigir muitos anos de tratamento, já que as recidivas são frequentes após as cirurgias ou após qualquer tratamento. Entre os novos tratamentos médicos, embora ainda não disponível no Brasil, está a administração oral de um novo progestagênio, Dienogest (DNG), o qual tem tido grande êxito no tratamento da endometriose, principalmente no tocante aos sintomas de dor associados. Esta revisão mostra os estudos mais relevantes realizados em diversos países com tratamentos em curto prazo - em geral 24 semanas - e em longo prazo - aproximadamente 53 semanas - e que mostraram alta eficácia, poucos eventos adversos e de pouca intensidade e alta satisfação das pacientes. A revisão também discute a relevância do DNG na prática clínica

Endometriosis is a chronic disease which affects young women at reproductive age and provokes pain, dyspareunia, infertility and which impaires patients' quality of life. Several treatments are available including medical and surgical. However, this disease must be considered as a chronic disease which could need many years of treatment because recurrences are common after surgery or any medical treatment. Among the new medical treatments, albeit not available in Brazil at the present time, there is the oral administration of a new progestin, Dienogest (DNG) which was successful in the treatment of endometriosis, mainly regarding pain-associated endometriosis. This review shows the main studies conducted in several countries with short term treatments - up to 24 weeks - as well as long-term treatments - in general up to 53 weeks - that showed high efficacy, few adverse events, and of middle or minimum intensity with high satisfaction of the patients. The review also discusses the relevance of DNG in the clinical practice

Telomerase activity alterations in sequential passages of mouse embryonic stem cells.

Telomerase is associated with cell proliferation capacity, protection and stabilization of chromosomes. TA (telomerase activity) can be detected in highly replicative cells, e.g. stem and cancer cells. Most available mESC (mouse embryonic stem cell) research is done with a few cell lines. The purpose of this study has been to evaluate the TA in different passages of newly isolated mESC. TRAP (Telomeric Repeat Amplification Protocol)-ELISA method was used in a semi-quantitative evaluation of TA. Three mESC lineages were investigated (CT2, CT3 and CT4) at three different passages (P13, P15 and P19). In contrast with previous studies, these mESC lines did not show the same TA throughout their passages, having initially low TA values, followed by a subsequent rise and stabilization.

Development and prevention of postsurgical adhesions in a chimeric mouse model of experimental endometriosis.

OBJECTIVE: To examine the impact of a recent surgery on development of endometriosis-related adhesions in a chimeric model and to determine the therapeutic efficacy of pioglitazone (PIO). DESIGN: Human endometrial biopsies were maintained in E(2) with or without PIO for 24 h before intraperitoneal injection into immunocompromised mice also treated with or without PIO at multiple time points after peritoneal surgery. The presence and extent of adhesions were examined in animals relative to the initial establishment of experimental endometriosis. SETTING: Medical school research center. PATIENT(S): Endometrial biopsies for experimental studies were provided by normally cycling women without a medical history indicative of endometriosis or adhesions. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Examination of the development of endometriosis-related adhesions in an experimental model. RESULT(S): Without therapeutic intervention, injection of E(2)-treated human endometrial tissue into mice near the time of peritoneal surgery resulted in multiple adhesions and extensive endometriotic-like disease. In contrast, PIO treatment reduced adhesive disease and experimental endometriosis related to surgical injury. CONCLUSION(S): The presence of human endometrial tissue fragments in the peritoneal cavity of mice with a recent surgical injury promoted development of both adhesive disease and experimental endometriosis. Targeting inflammation and angiogenesis with PIO therapy limited the development of postsurgical adhesions associated with ectopic endometrial growth.

Células-tronco: a resposta para os enigmas da patogênese da endometriose / Stem cells: the answer to the puzzles of the pathogenesis of endometriosis

A endometriose é uma doença crônica caracterizada pela presença de tecido endometrial ectópico. Esta doença frequentemente resulta em alta morbidade, incluindo dor pélvica crônica e infertilidade. A causa da endometriose é provavelmente multifatorial, sendo ainda objeto de muitos estudos. Fatores genéticos, ambientais e imunes estão possivelmente envolvidos na etiopatogenia desta doença, sendo especulada a associação com outros mecanismos. Nos últimos anos, alguns trabalhos têm demonstrado a presença de células-tronco/progenitoras no endométrio sadio; tais células possivelmente estão envolvidas na capacidade regenerativa desse tecido, assim como na patogênese de doenças ginecológicas proliferativas, como endometriose e carcinoma endometrial. Esta revisão avaliou as evidências disponíveis sobre a existência de células tronco/progenitoras endometriais e agrupou os resultados em uma hipótese de envolvimento dessas células na patogênese da endometriose. Foram selecionados os artigos mais relevantes sobre o tema. A identificação de células-tronco nos tecidos humanos e animais é presumida a partir da identificação de label retaining cells, células side population, marcadores de indiferenciação, potencial de clonogenicidade e diferenciação celular. A presença de supostas células-tronco no endométrio normal e ectópico foi demonstrada por alguns pesquisadores. Células endometriais no endométrio eutópico e em implantes endometrióticos, originadas a partir de células-tronco derivadas de medula óssea transplantada em humanos e de animais, também foram identificadas. Esses resultados sugerem que as células-tronco/progenitoras podem estar envolvidas na gênese da endometriose. No entanto, mais estudos são necessários para corroborar esta hipótese.

Endometriosis is a chronic disease characterized by the presence of ectopic endometrial tissue. This disease often results in high degree of morbidity, including chronic pelvic pain and infertility. Probably, the cause of endometriosis is multifactorial and it has been the object of many studies. Genetic, environmental and immune factors are possibly involved in the pathogenesis of this disease, and it is speculated that there are other mechanisms associated. In recent years, some studies have shown the presence of adult stem cells in the healthy endometrium. These cells are possible involved in the regenerative capability of endometrium and in the pathogenesis of proliferative gynecological diseases, such as endometriosis and endometrial carcinoma. This review evaluated the available evidence on the existence of stem/progenitor cells in the endometrium and gathered the results in an hypothesis of involvement of these cells in the pathogenesis of endometriosis. The most relevant articles about this subject were selected. The identification of stem cells in animal and human tissues is presumed from the identification of label retaining cells, side population cells, undifferentiation markers, besides the potential of clonogenesis and cellular differentiation. The presence of putative stem cells in the normal and ectopic endometrium was demonstrated by some researches. Endometrial cells in eutopic endometrium and endometriotic implants, originated from bone marrow-derived stem cells transplanted into humans and animals, have also been identified. These results suggest that stem/progenitor cells may be involved in the genesis of endometriosis. However, more studies are necessary to this hypothesis

O significado do grau de recomendaçãoe força de evidência A da classificaçãoda Associação Médica Brasileira

A revista Femina vem utilizando os graus de recomendações e força de evidência da Associação Médica Brasileira (AMB)1 para classificar as referências bibliográficas contidas no texto de seus artigos com o objetivo de dar ao leitor o verdadeiro valor atribuído a cada intervenção utilizada nas investigações

Quinacrine sterilization for human immunodeficiency virus-positive women.

OBJECTIVE: To evaluate the safety of nonsurgical quinacrine sterilization for HIV-positive (HIV+) women. DESIGN: An open trial of quinacrine sterilization was carried out in women infected with HIV and women who were HIV negative (HIV-). Comparison of the results with the two groups provided an assessment of the safety and effectiveness of quinacrine sterilization for HIV+ women. SETTING: University Medical School outpatient services. PATIENT(S): A total of 258 women who desired sterilization were offered quinacrine sterilization as a means of limiting family size. Sixty-four were HIV+, and 194 were HIV-. Women who were HIV+ had CD4 counts >200 and were otherwise healthy. INTERVENTION(S): A modified Copper T intrauterine device inserter was used to place 252 mg of quinacrine, divided into seven pellets (36 mg each) into the uterine cavity. Three insertions of this formulation were performed, 1 month apart. Viral load and CD8 and CD4 lymphocytes were measured both before and after quinacrine sterilization and at follow-up visits. Pregnancies and adverse events were recorded carefully. A decrement life table was made to statistically analyze results. RESULT(S) AND MAIN OUTCOME MEASURE(S): No serious adverse event occurred in any patient in this study. Adverse effects related to quinacrine sterilization were abdominal cramping, vulvar itching, nausea, and vaginal bleeding. Vaginal bleeding was the only short-term side effect noted to occur more frequently in HIV-infected women after quinacrine sterilization. Among HIV+ women, 35.9% had complaints of increased bleeding, whereas only 8.2% of those who were HIV- had such complaints, which probably were insertion related. Viral load and the CD4+ and CD8+ lymphocyte measures displayed no statistically significant difference after quinacrine sterilization. CONCLUSION(S): Quinacrine sterilization is a safe method for the sterilization of HIV-infected women and has no short-term effect on the pathology of the disease.

Sonographic evaluation of bladder volume in diabetic children and adolescents compared with nondiabetic subjects.

OBJECTIVES: An increase in volume and evidence of postvoiding residuals are both encountered in the bladder of diabetic patients and can lead to urinary tract infections and impair renal function. Our objective was to compare the bladder volume of diabetic children and adolescents with that of nondiabetic subjects. METHODS: We investigated 247 diabetic patients and 228 control subjects between the ages 3 and 21 years. The sonographic examination was performed with a full bladder, after spontaneous voiding, and after forced voiding. Three dimensions were determined: longitudinal, transverse, and anteroposterior transverse. The data were analyzed by the Chi2 test, analysis of variance, the Kruskal-Wallis test, and multivariate regression. RESULTS: The 2 groups were equivalent for age (P =.56) and sex (P =.82). The median full bladder volume was larger in the diabetic group (268 mL) than in the control group (220 mL; P =.0004). Postvoiding residual volume after spontaneous and forced voiding was significantly higher in the diabetic group (P <.0001). Multivariate analysis showed that being diabetic (P =.021), older than 9 years (P =.000), and female (P =.036) all influenced full bladder volume. CONCLUSIONS: Sonographic evaluation showed incipient bladder dysfunction in diabetic patients. The multivariate analysis showed that the increase in bladder volume had a association with age (>9 years), female sex, and mean glycosylated hemoglobin value. Thus, bladder volume evaluation by sonography should be incorporated in the routine assessment of patients with type 1 diabetes.

Avaliação da aplicabilidade da técnica de maturaçäo in vitro de oócitos humanos e posterior fertilização / Evaluation of the usefulness of the in vitro maturation technique of human oocyte and subsequent fertilization

OBJETIVO: avaliar a aplicabilidade da técnica de maturaçäo in vitro de oócitos humanos e posterior fertilizaçäo. MÉTODOS: estudo prospectivo näo randomizado descritivo realizado no período de novembro de 1999 a março de 2001 no qual foram incluídas 15 pacientes com infertilidade tubária e 20 ciclos de fertilizaçäo in vitro. Todas assinaram o termo de consentimento livre e esclarecido antes de iniciar o estudo. As pacientes tinham idade entre 18 e 32 anos incompletos, obstruçäo tubária como causa exclusiva de infertilidade e índice de massa corporal inferior a 25 kg/m². As pacientes receberam 300 UI de hormônio folículo estimulante (FSH) recombinante por via intramuscular no segundo dia do ciclo e doses adicionais de 150 UI no quarto e no sexto dia do ciclo. A coleta ovular foi realizada no sétimo dia do ciclo. Os oócitos foram colocados em meio TCM 199 acrescido de antibióticos, piruvato, FSH, gonadotrofina coriônica humana e soro (Serum Substitute Supplement - Irvine Scientific®). Após 48 h de cultivo, os oócitos que atingiram o estágio de metáfase II foram inseminados e os fertilizados foram transferidos. RESULTADOS: foram puncionados 144 folículos com a coleta de 67 oócitos imaturos (46,5 por cento). Quarenta e três oócitos atingiram o estágio de metáfase II (64,2 por cento) e foram inseminados. Destes, 30 fertilizaram e 25 embriöes foram transferidos para 10 pacientes. Houve uma gravidez com nascimento de um bebê. CONCLUSÄO: concluiu-se que a técnica de maturar oócitos humanos in vitro previamente à fertilizaçäo in vitro é técnica exeqüível, capaz de gerar gravidez