Scaffolds Vasculares Descelularizados Provenientes de Vasos Sanguíneos de Placenta Bovina / Decellularized Vascular Scaffolds Derived from Bovine Placenta Blood Vessels

Resumo Fundamento As doenças associadas ao aparelho circulatório são as principais causas de morbidade e mortalidade em todo o mundo, implicando a necessidade de implantes vasculares. Assim, a produção de biomateriais vasculares tem se mostrado uma alternativa promissora às terapias utilizadas em estudos e pesquisas relacionados à fisiologia vascular. Objetivos O presente projeto visa ao desenvolvimento artificial de vasos sanguíneos pela recelularização de scaffolds vasculares derivados de vasos placentários bovinos. Métodos A superfície corioalantoide da placenta bovina foi utilizada para produzir biomateriais descelularizados. Para a recelularização, 2,5 x 104 células endoteliais foram semeadas acima de cada fragmento de vaso descelularizado durante três ou sete dias, quando a cultura foi interrompida e os fragmentos foram fixados para análise de adesão celular. Biomateriais descelularizados e recelularizados foram avaliados por histologia básica, microscopia eletrônica de varredura e imuno-histoquímica. Resultados o processo de descelularização produziu vasos que mantiveram a estrutura natural e o conteúdo de elastina, e não foram observadas células e gDNA remanescentes. Além disso, células precursoras endoteliais se ligaram ao lúmen e à superfície externa do vaso descelularizado. Conclusão nossos resultados mostram a possibilidade de usos futuros desse biomaterial na medicina cardiovascular, como, por exemplo, no desenvolvimento de vasos artificiais.

Background Diseases associated with the circulatory system are the main causes of worldwide morbidity and mortality, implying the need for vascular implants. Thus, the production of vascular biomaterials has proven to be a promising alternative to therapies used in studies and research related to vascular physiology. Objectives The present project aims to achieve the artificial development of blood vessels through the recellularization of vascular scaffolds derived from bovine placental vessels. Methods The chorioallantoic surface of the bovine placenta was used to produce decellularized biomaterials. For recellularization, 2.5 x 104 endothelial cells were seeded above each decellularized vessel fragment during three or seven days, when culture were interrupted, and the fragments were fixed for cell attachment analysis. Decellularized and recellularized biomaterials were evaluated by basic histology, scanning electron microscopy, and immunohistochemistry. Results The decellularization process produced vessels that maintained natural structure and elastin content, and no cells or gDNA remains were observed. Endothelial precursor cells were also attached to lumen and external surface of the decellularized vessel.Conclusion: Our results show a possibility of future uses of this biomaterial in cardiovascular medicine, as in the development of engineered vessels.

Ursolic acid increases strength in mdx mice model and may decrease fibrosis deposition by tgf-ß downregulation / El ácido ursólico aumenta la resistencia en el modelo de ratones mdx y podría disminuir la deposición de fibrosis mediante la regulación decreciente de tgf-ß

SUMMARY: Dystrophin disfunction results in sarcolemma destabilization, leading muscle cell damage by continuous degeneration cycles and limited regeneration. In muscle dystrophy, caused by dystrophin dysfunction, inflammation, necrosis and fibrosis are pathophysiological muscle function loss characteristics. As a genetic disease, this muscle dystrophy has no cure, however, advances in drug therapy using glucocorticoids can decrease the disease progression. Subsequently, alternative therapies were studied, such as ursolic acid (UA), that inhibits muscle atrophy and increases muscle mass and strength. Herein, we used 10 mg/kg daily supplementation in mdx mice for 4 weeks to evaluate serum creatine phosphokinase (CPK), muscle strength (Kondziela test), muscular organization (histology) and expression of fibrosis related genes (TGF-ß, TNF-α, mstn and ostn). UA supplementation increased muscle morphological organization, motor strength and decreased muscular TGF-ß expression. Altogether, the gene expression profile, histological organization and strength could suggest that UA treatment did not stop the fibrogenesis but decreased its progress.

RESUMEN: La disfunción de la distrofina resulta en la desestabilización del sarcolema, llevando al daño de las células musculares por ciclos continuos de degeneración y regeneración limitada. En la distrofia muscular, debido a la disfunción de la distrofina, la inflamación, la necrosis y la fibrosis, son características fisiopatológicas de la pérdida de la función muscular. Como enfermedad genetica no es possible remediar esta distrofia muscular, sin embargo, los avances en la terapia de medicamentos con glucocorticoides pueden disminuir la progresión de la enfermedad. Se estudiaron terapias alternativas, como el ácido ursólico (UA), que inhibe la atrofia muscular y aumenta la masa y la fuerza muscular. En este estudio, utilizamos una suplementación diaria de 10 mg / kg en ratones mdx durante 4 semanas para evaluar la creatina fosfoquinasa (CPK) sérica, la fuerza muscular (prueba de Kondziela), la organización muscular (histología) y la expresión de genes relacionados con la fibrosis (TGF-ß, TNF- α, mstn y ostn). La suplementación con AU aumentó la organización morfológica muscular, la fuerza motora y la disminución de la expresión muscular de TGF-ß. El perfil de expresión génica, la organización histológica y la fuerza simultáneamente podrían sugerir que el tratamiento con AU no detuvo la fibrogénesis sino que disminuyó su progreso.

Equine yolk sac: a stem cells source / Saco vitelino equino: una fuente de células madre

SUMMARY: Mesenchymal stem cells are characterized by in vitro high proliferation and multilineage potential maintenance. This study aimed to isolate and characterize equine YS mesenchymal stem cells and compare these with amniotic membranes. The yolk sac (YS) and amniotic membranes (AM) were obtained from 20 pregnant mares with gestational age around 30 days. Cells were cultured in α-MEM supplemented with 15 % FBS, 1 % antibiotic solution, 1 % L-glutamine and 1 % nonessential amino acids. To cell characterization we used cytogenetic analysis, fibroblast colony-forming unit assays, cell growth curves, immunophenotyping, flow cytometry, differentiation assays and teratoma formation. Results: Both cell sources presented fibroblastoid and epithelioid-like format. The YS cells have lower colony formation potential then AM ones, 3 versus 8 colonies per 103 plated cells. However, YS cells grew progressively while AM cells showed steady. Both, the YS and amnion cells immunolabeled for Oct-4, Nanog, SSEA-3, cytokeratin 18, PCNA, and vimentin. In addition, presented mesenchymal, hematopoietic, endothelial and pluripotency markers in flow cytometry. Discussion: Both cell sources presented high plasticity and differed into osteogenic, adipogenic, and chondrogenic lineages, and no tumor formation in nude mice was observed. The results suggest that horse YS may be useful for cell therapy such as amnion-derived cells.

RESUMEN: Las células madre mesenquimales se caracterizan por una alta proliferación in vitro y un mantenimiento potencial de múltiples líneas. Este estudio tuvo como objetivo aislar y caracterizar las células madre mesenquimales del saco vitelino equinas y compararlas con las membranas amnióticas. Se obtuvo el saco vitelino (SV) y las membranas amnióticas (MA) de 20 yeguas preñadas con edad gestacional de aproximadamente 30 días. Las células se cultivaron en α -MEM suplementado con 15 % de FBS, 1 % de solución antibiótica, 1 % de L-glutamina y 1 % de aminoácidos no esenciales. Para la caracterización celular utilizamos análisis citogenéticos, ensayos de unidades de colonias de fibroblastos, curvas de crecimiento celular, inmunofenotipaje, citometría de flujo, ensayos de diferenciación y formación de teratomas. Ambas fuentes celulares presentaron formato fibroblastoideo y epitelioide. Las células SV tienen un potencial de formación de colonias más bajo que las de MA, 3 versus 8 colonias por 103 células en placa. Sin embargo, las células SV crecieron progresivamente mientras que las células MA se mostraron estables. Tanto las células YS como las células amnios están inmunomarcadas para Oct-4, Nanog, SSEA-3, citoqueratina 18, PCNA y vimentina. Además, presentó marcadores mesenquimales, hematopoyéticos, endoteliales y pluripotenciales en citometría de flujo. Ambas fuentes celulares presentaron alta plasticidad y diferían en linajes osteogénicos, adipogénicos y condrogénicos, y no se observó formación de tumores en ratones. Los resultados sugieren que el SV de caballo puede ser útil para la terapia celular, como las células derivadas de amnios.

Rabbit vomeronasal organ-derivered cells have mesenchymal profile and neuronal commitment / Las células derivadas del órgano vomeronasal del conejo tienen perfil mesenquimatoso y compromiso neuronal

SUMMARY: The vomeronasal organ (VNO) is an accessory organ involved on the olfactory pathway, that detects pheromones and emits signals in order to modulate social and reproductive behavior. The VNO stem cells replace neurons throughout life. The aim of this study was to isolate and characterize cells derived from the vomeronasal organ from New Zealand rabbits. Five male rabbits with 120 days were used for cell isolation and culture. Results: VNO-derived cells presented labelling for proliferation (PCNA), undifferentiated profile (Nanog), neuronal (GFAP), mesenchymal stem cells (CD73, CD90 and CD105 and Stro-1). Also, presence of cytoskeletal (Vimentin, b-tubulin and CK-18) and absence of hematopoietic markers (CD34, CD117 and CD45) both by immunofluorescence and flow cytometry. By PCR it was possible to verify the expression of some undifferentiated profile (Oct-4), neuronal (Nestin) and mesenchymal (CD73, CD105 and Vimentin) genes. Functionally, VNO-derived cells differentiate in vitro into adipocytes, osteocytes and chondrocytes, and presented no tumorigenic potential when injected to Balb/c nu/nu mice. In conclusion, the rabbit VNO-derived cells have a profile that could be supportive to VNO olfactory/neuroreceptor epithelium by delivering factors to epithelial turnover or even by differentiation into epithelial cells to replacement of commissural epithelium.

RESUMEN: El órgano vomeronasal (OVN) es un órgano accesorio de la vía olfatoria, que detecta feromonas y emite señales que afectan la modulación del comportamiento social y reproductivo. Las células madre OVN reemplazan las neuronas durante toda la vida. El objetivo de este estudio fue aislar y caracterizar células derivadas del órgano vomeronasal de conejos raza Nueva Zelanda. Para el aislamiento y el cultivo celular se utilizaron cinco conejos machos con una edad de 120 días. Las células del OVN presentaron etiquetado para la proliferación (PCNA), un perfil indiferenciado (Nanog), neuronal (GFAP), células madre mesenquimales (CD73, CD90 y CD105 y Stro-1). Además, se ob- servó presencia de citoesqueleto (Vimentina, β-tubulina y CK-18) y ausencia de marcadores hematopoyéticos (CD34, CD117 y CD45) tanto por inmunofluorescencia como por citometría de flujo. Me- diante PCR fue posible verificar la expresión de algunos genes de perfil indiferenciado (Oct-4), neuronal (Nestin) y mesenquimatoso (CD73, CD105 y Vimentin). Las células derivadas del OVN se diferencian in vitro en adipocitos, osteocitos y condrocitos, y no presentan un potencial tumorigénico al ser infiltrados en ratones Balb / c nu / nu. En conclusión, las células derivadas de OVN de conejo tienen un perfil que podría ser compatible con el epitelio olfatorio / neurorreceptor de OVN transmitiendo factores al recambio epitelial o incluso mediante la diferenciación en células epiteliales para reemplazar el epitelio comisural.

Characteristics of the equine placenta at first trimester / Características de la placenta equina en el primer trimestre

The equine placenta is a simple apposition of fetal and maternal tissues, becoming more complex with the formation of microcotyledons around days 75 and 100 of gestation. The present study aimed to describe the gross and microscopic morphology of early equine placenta. Embryonic/fetal membranes from thirty-seven mares were submitted to macroscopic description, light, scanning and transmission microscopy. Overall the gross characteristics of membranes were similar with already described for older stages. However, transmission electron microscopy evidenced high metabolic rate in chorion and allantois, and high secretion profile in amnion and even higher in yolk sac. Gene ontologies enrichment, using published data, pointed several common ontologies in allantoic and amniotic fluids, related to oxygen and iron transport, extracellular space and high-density lipoprotein receptor binding. Overall, the morphological and ontology enrichment could indicate allantois and amnion crosstalk.

La placenta equina es una simple aposición de tejidos fetales y maternos, que se vuelve más compleja con la formación de microcotiledones alrededor de los días 75 y 100 de gestación. El presente estudio tuvo como objetivo describir la morfología macroscópica y microscópica de la placenta equina temprana. Las membranas embrionarias / fetales de treinta y siete yeguas fueron sometidas a descripción macroscópica, luz, escaneo y microscopía de transmisión. En general, las características generales de las membranas fueron similares a las ya descritas para las etapas más antiguas. Sin embargo, la microscopía electrónica de transmisión mostró una alta tasa metabólica en corion y alantoides, y un alto perfil de secreción en amnios e incluso mayor en el saco vitelino. El enriquecimiento de ontologías génicas, utilizando datos publicados, señaló varias ontologías comunes en fluidos alantoideos y amnióticos, relacionados con el transporte de oxígeno y hierro, espacio extracelular y unión a receptores de lipoproteínas de alta densidad. En general, el enriquecimiento morfológico y ontológico podría indicar alantoides y diafonía de amnios.

New oropharyngeal double lumen cannula for sedation for transesophageal echocardiography: case series / Nova cânula orofaríongea de duplo lúmen para sedação em ecocardiografia transesofágica: série de casos

Abstract Introduction: Currently, transesophageal echodopplercardiography is frequently performed under sedation on an outpatient basis. Sedation is related with increase in incidents on airways. Bearing in mind this scenario, we developed a new double lumen oropharyngeal cannula aimed at keeping airway patency, in addition to reducing risks to patients during endoscopy procedures performed under sedation. The main objective of our study was to assess the incidence of oxygen desaturation in a series of cases of adult patients submitted to outpatient transesophageal echo exam, under sedation and using the oropharyngeal cannula. Method: 30 patients under sedation with intravenous midazolam and propofol were assessed. After loss of consciousness, the cannula was placed and patients were maintained on spontaneous breathing. Oxygen saturation, capnometry, heart rate and non-invasive arterial blood pressure, in addition to subjective data: airway patency, handling of cannula insertion and, comfort of examiner were analyzed. Results: The incidence of mild desaturation was 23.3%, and there was no severe desaturation in any of the cases. The insertion of the oropharyngeal cannula was considered easy for 29 patients (96.6%), and transesophageal echo probe handling was appropriate in 93.33% of exams performed. Conclusions: Transesophageal echo exams under sedation aided by the double-lumen oropharyngeal cannula presented a low incidence of desaturation in patients assessed, and allowed analysis of expired CO2 during the exams.

Resumo Introdução: Nos dias atuais, exames de ecocardiografia transesofágica (ETE) são realizados de forma frequente sob sedação em regime ambulatorial. A sedação está relacionada com aumento de intercorrências nas vias aéreas. Dentro desse contexto, desenvolvemos uma cânula orofaríngea de duplo-lúmen com finalidade de manutenção da via aérea pérvia, reduzindo riscos aos pacientes durante procedimentos endoscópicos sob sedação. O principal objetivo do nosso estudo foi avaliar a incidência de dessaturação em uma série de casos de pacientes adultos submetidos a ETE ambulatorial sob sedação com o uso da cânula orofaríngea. Métodos: Foram avaliados 30 pacientes sedados com midazolam e propofol intravenoso. A cânula foi inserida após perda da consciência e os pacientes foram mantidos com ventilação espontânea. Analisados saturação de oxigênio, capnometria, frequência cardíaca e pressão arterial não invasiva, além de dados subjetivos: patência das vias aéreas, manuseio e inserção da cânula e conforto ao examinador. Resultados: A incidência de dessaturação leve foi de 23.3% e não houve dessaturação grave em nenhum caso. A inserção da cânula orotraqueal foi considerada fácil em 29 pacientes (96,6%) e o manuseio da sonda de ETE foi adequada em 93,33% dos exames realizados. Conclusões: A realização dos exames de ETE sob sedação com auxílio da cânula orofaríngea de duplo lúmen apresentou baixa incidência de dessaturação nos pacientes avaliados, além de permitir análise do CO2 expirado durante a realização dos exames.

Comparação entre Técnicas de Block-Matching e Optical Flow na análise do Strain pela Ecocardiografia / Comparison of Block-Matching and Optical Flow in Strain Echocardiography Analysis

Introdução: Diferentes tecnologias têm sido utilizadas para avaliar a função sistólica do ventrículo esquerdo. De particular interesse, está a ecocardiografia com strain bidimensional (2DSTE). Dois métodos diferentes têm sido usados para quantificar a deformação miocárdica (strain miocárdico) pela 2DSTE: block matching e optical flow. Ambos estão presentes em ecocardiógrafos comercialmente disponíveis. Entretanto, não há consenso a respeito se as medidas do strain longitudinal por estes métodos são sobreponíveis. Objetivo: Comparar os valores de pico do strain miocárdico longitudinal obtidos através da 2DSTE pelos dois diferentes métodos (block matching x optical flow). Método: Foram realizadas as medidas do strain em 16 segmentos miocárdicos do ventrículo esquerdo, conforme orientação da Sociedade Americana de Ecocardiografia pela técnica do block matching (Vivid 7, GE, Horten, Noruega) e, imediatamente após, pela técnica do optical flow (My Lab 60, Esaote, Florença, Itália) de forma randomizada e seus valores foram comparados. Resultados: Houve 28 indivíduos com idade 27,9 ± 7,7 anos, sendo 50% do sexo masculino e todos com fração de ejeção superior a 55%. A medida do strain longitudinal global foi maior pela técnica de block matching (p = 0,02). Na análise dos 16 segmentos, houve 10 segmentos com valores diferentes, em especial os segmentos apicais. Conclusão: Os valores do strain miocárdico longitudinal obtidos por métodos diferentes não são sobreponíveis e devem ser usados com cautela. Sendo assim, valores de normalidade também variam de acordo com o fabricante e o método utilizado

Introduction: Different technologies have been used to evaluate left ventricular systolic function. Of particular interest is the two-dimensional strain echocardiography (2DSTE). Two different methods have been used to quantify myocardial strain by 2DSTE: block matching and optical flow. Both are present in commercially available echocardiographs. However, there is no consensus as to whether the longitudinal strain measures using these methods are overlapping. Objective: To compare the longitudinal myocardial strain peak values obtained through 2DSTE using two different methods (block matching x optical flow). Method: Strain measurements in 16 left ventricular myocardial segments were taken as recommended by the American Society of Echocardiography using block matching (Vivid 7, GE, Horten, Norway) and, immediately after, using optical flow (My Lab 60, Esaote, Florence, Italy) in a randomized way and their values were compared. Results: There were 28 individuals aged 27.9 ± 7.7, of whom 50% were male and all with an ejection fraction greater than 55%. Global longitudinal strain was higher using block matching (p = 0.02). In the analysis of the 16 segments, 10 segments had different values, especially the apical segments. Conclusion: Longitudinal myocardial strain values obtained by different methods are not overlapping and must be used with caution. Normality values also vary according to the manufacturer and the method used

The Brazilian version of the Constant-Murley Score (CMS-BR): convergent and construct validity, internal consistency, and unidimensionality / Versão brasileira do Constant-Murley Score (CMS-BR): validade convergente e de constructo, consistência interna e unidimensionalidade

ABSTRACT OBJECTIVES: To translate and culturally adapt the CMS and assess the validity of the Brazilian version (CMS-BR). METHODS: The translation was carried out according to the back-translation method by four independent translators. The produced versions were synthesized through extensive analysis and by consensus of an expert committee, reaching a final version used for the cultural adaptation. A field test was conducted with 30 subjects in order to obtain semantic considerations. For the psychometric analyzes, the sample was increased to 110 participants who answered two instruments: CMS-BR and the Disabilities of the Arm, shoulder and Hand (DASH). The CMS-BR and DASH score range from 0 to 100 points. For the first, higher points reflect better function and for the latter, the inverse is true. The validity was verified by Pearson's correlation test, the unidimensionality by factorial analysis, and the internal consistency by Cronbach's alpha. RESULTS: The explained variance was 60.28% with factor loadings ranging from 0.60 to 0.91. The CMS-BR exhibited strong negative correlation with the DASH score (-0.82, p < 0.05), Cronbach's alpha 0.85, and its total score was strongly correlated with the patient's range of motion (0.93, p < 0.001). CONCLUSION: The CMS was satisfactorily adapted for Brazilian Portuguese and demonstrated evidence of validity that allows its use in this population.

RESUMO OBJETIVOS: Traduzir e adaptar culturalmente o Constant-Murley Score (CMS) e verificar a validade da versão brasileira (CMS-BR). MÉTODOS: A tradução foi realizada de acordo com o método de retrotradução por quatro tradutores independentes. As versões produzidas foram sintetizadas por análise extensiva e consenso de um comitê de especialistas, gerando uma versão final usada para a adaptação cultural. Realizou-se um teste em campo com 30 sujeitos para observação de possíveis considerações em relação à semântica. Para a posterior análise psicométrica, ampliou-se a amostra para 110 participantes, que responderam a dois instrumentos: CMS-BR e Disabilities of the Arm, Shoulder and Hand (DASH). O CMS-BR e o DASH possuem variação de 0 a 100 pontos. Para o primeiro, altas pontuações refletem melhor função, para o segundo, o contrário. A validade foi verificada através do teste de correlação de Pearson, a dimensionalidade através de análise fatorial e a consistência interna através do alfa de Cronbach. RESULTADOS: A variância explicada foi de 60,28% com cargas fatoriais entre 0,60 e 0,91. O CMS-BR demonstrou correlação forte e negativa com o DASH (-0,82, p < 0,05), alfa de Cronbach de 0,85 e seu escore total teve correlação forte com a amplitude de movimento dos pacientes (0,93, p < 0,001). CONCLUSÃO: O CMS-BR foi adaptado de forma satisfatória e demonstrou evidências de validade que permitem seu uso nessa população.

Global Postural Reeducation for patients with musculoskeletal conditions: a systematic review of randomized controlled trials

ABSTRACT Objectives To systematically review randomized controlled trials that assessed the effects of Global Postural Reeducation (GPR) on patient-reported outcomes in conditions of the musculoskeletal system. Method An electronic search of MEDLINE (via PubMed), EMBASE, Cochrane CENTRAL, and SciELO was performed from their inception to June 2015. Randomized controlled trials that analyzed pain and patient-reported outcomes were included in this review. The Cochrane Collaboration’s Risk of Bias Tool was used to evaluate risk of bias, and the quality of evidence was rated following the GRADE approach. There were no language restrictions. Results Eleven trials were included totaling 383 patients. Overall, the trials had high risk of bias. GPR was superior to no treatment but not to other forms of treatment for pain and disability. No placebo-controlled trials were found. Conclusion GPR is not superior to other treatments; however, it is superior to no treatment. Due to the lack of studies, it is unknown if GPR is better than placebo. The quality of the available evidence ranges from low to very low, therefore future studies may change the effect estimates of GPR in musculoskeletal conditions.

Ultrastructural analysis of the spermatogenesis in the guinea pig (Cavia porcellus) / Análise ultraestrutural da espermatogênese do porquinho-da-Índia (Cavia porcellus)

Understanding of the reproductive function is essential for both, the establishment of appropriate management systems, and for the use of new species as animal models. In this study, we used light and electron microscopy to characterize the sexual development stages of the guinea pig (Cavia porcellus) in specimens of 30, 45 and 90 days of age. We observed the differentiation of spermatocytes only through transmission electron microscopy in the leptotene, zygotene and pachytene phases of meiosis, in 30-day-old animals. During puberty, there was differentiation of the germinative epithelium and formation of the acrosome. Spermatozoa, however, were not detected. Thus, we could infer that puberty happens after 45 days of age. Sexual maturity was evident in 90-day-old specimens. Our results showed that changes in the testicular germinative epithelium during the postnatal sexual development in guinea pig led to morphological changes, including the ones related to the development of Leydig and Sertoli cells, which are directly related to puberty. In this work, we provide new morphological subsidies for a better understanding of reproductive parameters of this species, enabling its use as an animal model in the field of the reproductive biology.(AU)

A função reprodutiva é um fator de vital compreensão tanto para o estabelecimento de sistemas apropriados de manejo, quanto para o uso de novas espécies como modelos animais. Neste estudo através da microscopia de luz e eletrônica caracterizou-se a fase de desenvolvimento sexual do porquinho-da-Índia (Cavia porcellus) em espécimes de 30, 45 e 90 dias de desenvolvimento. Nos animais de 30 dias, a diferenciação dos espermatócitos foi visualizada somente na microscopia eletrônica de transmissão em leptóteno, zigóteno e paquíteno. Durante a puberdade, houve diferenciação do epitélio germinativo, formação do acrossoma, porém não foram evidenciados espermatozóides, assim, infere-se que a puberdade acontece a partir dos 45 dias de idade. A maturidade sexual foi evidente aos 90 dias de idade. Nossos resultados mostraram que ao longo do desenvolvimento sexual pós-natal do porquinho-da-Índia, mudanças no epitélio germinativo testicular levam há alterações morfológicas, inclusive com relação ao desenvolvimento das células de Sertoli e de Leydig, as quais estão diretamente relacionadas com a puberdade. Assim, novos subsídios morfológicos são fornecidos para um melhor entendimento dos parâmetros reprodutivos desta espécie, a fim de viabilizar sua utilização como modelo animal no campo da biologia reprodutiva.(AU)

The effects of whole body vibration in patients with type 2 diabetes: a systematic review and meta-analysis of randomized controlled trials

Background: Whole body vibration (WBV) has been used to increase physical activity levels in patients with type 2 diabetes mellitus (T2DM). Objective: To carry out a systematic review of the effects of WBV on the glycemic control, cardiovascular risk factors, and physical and functional capacity of patients with T2DM. Method: MEDLINE, LILACS, PEDro, and Cochrane Central Register of Controlled Trials were searched up to June 1st, 2015. Randomized controlled trials investigating the effects of WBV, compared to control or other intervention, on blood glucose levels, blood and physical cardiovascular risk factors, and physical and functional capacity in adult individuals with T2DM. Two independent reviewers extracted the data regarding authors, year of publication, number of participants, gender, age, WBV parameters and description of intervention, type of comparison, and mean and standard deviation of pre and post assessments. Results: Out of 585 potentially eligible articles, two studies (reported in four manuscripts) were considered eligible. WBV interventions provided a significant reduction of 25.7 ml/dl (95% CI:-45.3 to -6.1; I2: 19%) in 12 hours fasting blood glucose compared with no intervention. Improvements in glycated hemoglobin, cardiovascular risk factors, and physical and functional capacity were found only at 12 weeks after WBV intervention in comparison with no intervention. Conclusion: WBV combined with exercise seems to improve glycemic control slightly in patients with T2DM in an exposure-dependent way. Large and well-designed trials are still needed to establish the efficacy and understand whether the effects were attributed to vibration, exercise, or a combination of both.

Rabbit olfactory stem cells. Isolation protocol and characterization

PURPOSE: To describe a new technique for isolation of a mesenchymal stem cells (MSCs) population from the olfactory mucosa in rabbits. METHODS: Olfactory stem cells (OSCs) were retrieved from under the cribriform plate of the Ethmoid bone. Several assays were accomplished to characterize the cell population and attest its viability in vitro. The cells were submitted to flow cytometry with the antibodies CD34, CD45, CD73, CD79, CD90 and CD105 and also they were induced to differentiate in three lineages. Functional evaluation involved analysis of in vitro growth behavior, colony forming unit like fibroblasts (CFU-f) and cryopreservation response. Further transduction with Green Fluorescent Protein (GFP) was also performed. RESULTS: The OSCs showed mesenchymal features, as positive response to CD34, CD73 and CD90 antibodies and plasticity. Additionally, these cells have high proliferated rate, and they could be cultured through many passages and kept the ability to proliferate and differentiate after cryopreservation. The positive response to the transduction signalizes the possibility of cellular tracking in vivo. This is a desirable feature in case those cells are used for pre-clinical trials. CONCLUSION: The cells harvested were mesenchymal stem cells and the technique described is therefore efficient for rabbit olfactory stem cells isolation.

Bile acids as potential pheromones in pintado catfish Pseudoplatystoma corruscans (Spix & Agassiz, 1829): eletrophysiological and behavioral studies

Bile acids are potent olfactory and gustatory stimulants for fish. Electro-olfactogram recording was used to test whether the olfactory epithelium of pintado catfish Pseudoplatystoma corruscans is specifically sensitive to bile acids, some of which have been hypothesized to function as pheromones. Five out of 30 bile acids that had been pre-screened for olfactory activity in fish were selected. Cross-adaptation experiments demonstrated that sensitivity to bile acids is attributable to at least 3 independent classes of olfactory receptor sites. The taurocholic acid (TCA) and taurochenodeoxycholic acid (TCD) were the most potent compounds. By using avoidance/preference tests, we found that P. corruscans prefers water containing TCA. Bile acids are discriminated by olfactory epithelium of pintado, supporting that these compounds could function as pheromones.

Os ácidos biliares são potentes estimulantes olfatórios e gustatórios em peixes. Registros em eletro-olfactograma foram usados para testar se o epitélio olfatório de Pseudoplatystoma corruscans, pintado, é sensível aos ácidos biliares, alguns dos quais têm sido propostos como feromônios. Foram selecionados cinco de uma lista de trinta ácidos biliares previamente testados em atividade olfatória em peixes. Testes de adaptação cruzada demonstraram que a sensibilidade aos ácidos biliares se dá por 3 classes independentes de sites de receptores olfatórios. O ácido taurocólico (TCA) e o ácido tauroquenodesoxicólico (TCD) foram os compostos mais potentes. Em testes de evasão/preferência, P. corruscans prefere água contendo o ácido TCA. Os ácidos biliares são discriminadas por epitélio olfatório de pintado, evidenciando que estes compostos podem funcionar como feromônios.

Impacto da Curva de Aprendizado na Seleção de Pacientes e nos Resultados Clínicos do Implante por Cateter de Prótese Aórtica / Impact of Transcatheter Aortic Valve Implantation Learning Curve on Patient Selection and Clinical Outcomes

Introdução: O implante por cateter de prótese aórtica (TAVI, do inglês transcatheter aortic valve implantation) constitui tratamento alternativo para pacientes com estenose aórtica de alto risco cirúrgico ou inoperáveis. Para adquirir competência, o grupo multidisciplinar deve receber treinamento específico e acumular experiência na execução do TAVI. Contudo, sua curva de aprendizado não está bem estabelecida. Nosso objetivo foi analisar o impacto da curva de aprendizado na seleção de pacientes, nos aspectos técnicos e nos resultados clínicos do TAVI. Métodos: Estudo observacional e prospectivo dos primeiros 150 pacientes submetidos a TAVI por via femoral, entre janeiro de 2009 e dezembro de 2013 divididos em tercis (n = 50) de acordo com a data do procedimento. Os desfechos foram definidos conforme os critérios Valve Academic Research Consortium-2 (VARC-2). Resultados: A idade foi de 82,5 ± 6,7 anos, sendo 44% homens e 75% em classe NYHA III/IV. O EuroSCORE (24,2 ± 13% vs. 21,2 ± 10,8% vs. 23,4 ± 14,3%) e o STS Score (5,9 ± 2,9% vs. 6,7 ± 4,3% vs. 5,8 ± 3,1%) foram similares entre os grupos. Observou-se redução gradativa nos tempos do procedimento (107,2 ± 48,1 minutos vs. 90,3 ± 42,2 minutos vs. 76,6 ± 37,7 minutos; p < 0,01) e de fluoroscopia (31,3 ± 9,6 minutos vs. 25,4 ± 8,7 minutos vs. 17,2 ± 6,2 minutos; p = 0,01), e no ...

Background: Transcatheter aortic valve implantation (TAVI) is an alternative treatment for high-risk or inoperable patients with aortic stenosis. The multidisciplinary team must undergo specific training and accumulate experience to achieve optimal results. However, its learning curve is not well established. Our objective was to investigate the impact of learning curve on patient selection, technical aspects and clinical outcomes of TAVI. Methods: Observational, prospective analysis of the first 150 patients undergoing transfemoral TAVI between January 2009 and December 2013. Patients were divided into tertiles (n = 50), according to the procedure date. Outcomes were defined according to Valve Academic Research Consortium-2 (VARC-2) criteria. Results: Mean age was 82.5 ± 6.7 years, 44% were male and 75% were in NYHA class III/IV. EuroS-CORE (24.2 ± 13% vs. 21.2 ± 10.8% vs. 23.4 ± 14.3%) and STS Score (5.9 ± 2.9% vs. 6.7 ± 4.3% vs. 5.8 ± 3.1%) were similar between groups. A gradual decrease was observed in procedure times (107.2 ± 48.1 minutes vs. 90.3 ± 42.2 minutes vs. 76.6 ± 37.7 minutes; p < 0.01), fluoroscopy times (31.3 ± 9,6 minutes vs. 25.4 ± 8.7 minutes vs. 17.2 ± 6.2 minutes; p = 0.01) and contrast volume (145.5 ± 70.9 mL vs. 123.2 ± 87.8 mL vs. 101.1 ± 50 mL; p = 0.01). Mortality decreased gradually (20% vs. 10% vs. 4%; p = 0,047), and lower bleeding and moderate-to-severe aortic regurgitation were ...

Análise da incidência e preditores clínicos e ecocardiográficos do refluxo paraprotético aórtico após o implante de prótese aórtica transcateter / Analysis of the incidence and clinical and echocardiographic predictors of paravalvular aortic regurgitation after transcatheter aortic valve implantation

INTRODUÇÃO: A incidência de refluxo paraprotético (RPP) parece maior entre os pacientes submetidos a implante de prótese aórtica transcateter e sua potencial associação com aumento da mortalidade tardia tem suscitado preocupação na comunidade científica. Nosso objetivo foi avaliar a incidência e o impacto clínico e estabelecer preditores do RPP em nossa casuística. MÉTODOS: Entre julho de 2009 e fevereiro de 2013, 112 pacientes foram submetidos a implante de prótese aórtica transcateter. O grau do RPP pós-procedimento foi avaliado segundo os critérios do VARC 2. Dividiu-se a população em grupo RPP ausente/RPP discreto e grupo RPP moderado/RPP grave. RESULTADOS: A média da idade foi de 82,5 ± 3,9 anos, 58,9% eram do sexo feminino e o EuroSCORE logístico foi de 23,6 ± 13,4. Houve queda do gradiente sistólico médio (54,7 ± 15,3 mmHg vs. 11,7 ± 4 mmHg; P < 0,01) e ganho da área valvar aórtica (0,66 ± 0,15 cm² vs. 1,8 ± 0,3 cm²; P < 0,01). Ao final do procedimento, 46,4% não apresentaram RPP, e RRP discreto ou moderado foi observado em 42% e 11,6% dos pacientes. Nenhum paciente apresentou RPP grave. A análise multivariada identificou sexo masculino [odds ratio (OR) 5,85, intervalo de confiança (IC] 1,29-26,7; P = 0,022), valvoplastia aórtica percutânea prévia (OR 18,44, IC 2,30-147,85; P = 0,006), fração de ejeção < 35% (OR 4,160, IC 1,014-17,064; P = 0,048) e presença de hipertensão pulmonar grave (OR 7,649, IC 1,86-31,51; P = 0,005) como preditores independentes de RPP moderado/grave. CONCLUSÕES: A incidência de RPP moderado/grave foi baixa e comparável à de outras casuísticas. Sexo masculino, antecedente de valvoplastia aórtica percutânea prévia, presença de hipertensão pulmonar grave e disfunção ventricular esquerda grave foram preditores independentes dessa complicação.

BACKGROUND: The incidence of paravalvular aortic regurgitation (PAR) seems higher among patients submitted to transcatheter aortic valve implantation and its potential association with an increased late mortality has raised concerns in the scientific community. Our objective was to evaluate the incidence and clinical impact of PAR and establish PAR predictors in our patient population. METHODS: Between July/2009 and February/2013, 112 patients were submitted to transcatheter aortic valve implantation. The degree of PAR after the procedure was assessed according to the VARC 2 criteria. The population was divided into no/mild PAR group and moderate/severe PAR group. RESULTS: Mean age was 82.5 ± 3.9 years, 58.9% were female and the logistic EuroSCORE was 23.6 ± 13.4. There was a decrease in the mean systolic gradient (54.7 ± 15.3 mmHg vs 11.7 ± 4 mmHg; P < 0.01) and a gain in the aortic valve area (0.66 ± 0.15 cm² vs 1.8 ± 0.3 cm²; P < 0.01). At the end of the procedure 46.4% did not have PAR, and mild or moderate PAR was observed in 42% and 11.6% of the patients. No patient presented severe PAR. Multivariate analysis identified male gender [odds ratio (OR) 5.85, confidence interval (CI] 1.29-26.7; P = 0.022), previous percutaneous aortic val­vuloplasty (OR 18.44, CI 2.30-147.85; P = 0.006), ejection fraction < 35% (OR 4.160, CI 1.014-17.064; P = 0.048) and the presence of severe pulmonary hypertension (OR 7.649, CI 1.86-31.51; P = 0.005) as independent predictors of moderate/severe PAR. CONCLUSIONS: The incidence of moderate/severe PAR was low and comparable to other studies. Male gender, history of prior percutaneous aortic valvuloplasty, presence of severe pulmonary hypertension and severe left ventricular dysfunction were independent predictors of this complication.

Correção do astigmatismo irregular com lente intraocular tórica em um paciente com catarata e degeneração marginal pelúcida: relato de caso / Toric intraocular lens implantation for cataract and irregular astigmatism related to pellucid marginal degeneration: case report

A degeneração marginal pelúcida (DMP) é uma rara ectasia corneana cuja progressão resulta em astigmatismo irregular e baixa visual não corrigidos com óculos ou lentes de contato. O presente relato descreve um paciente com catarata e DMP que foi tratado com facoemulsificação e implante de lente intraocular tórica com recuperação da acuidade visual em ambos os olhos.

Pellucid marginal degeneration (PMD) is a rare corneal ectasia and its progression leads to irregular astigmatism and low vision that can not have spectacles or contact lens correction. This report describes a patient with low vision due to cataract and PMD that was treated with phacoemulsification and implantation of a toric intraocular lens with a satisfactory visual acuity outcome.