Horizontal transfer of exosomal microRNAs transduce apoptotic signals between pancreatic beta-cells.

BACKGROUND: Diabetes mellitus is a common metabolic disorder characterized by dysfunction of insulin-secreting pancreatic beta-cells. MicroRNAs are important regulators of beta-cell activities. These non-coding RNAs have recently been discovered to exert their effects not only inside the cell producing them but, upon exosome-mediated transfer, also in other recipient cells. This novel communication mode remains unexplored in pancreatic beta-cells. In the present study, the microRNA content of exosomes released by beta-cells in physiological and physiopathological conditions was analyzed and the biological impact of their transfer to recipient cells investigated. RESULTS: Exosomes were isolated from the culture media of MIN6B1 and INS-1 derived 832/13 beta-cell lines and from mice, rat or human islets. Global profiling revealed that the microRNAs released in MIN6B1 exosomes do not simply reflect the content of the cells of origin. Indeed, while a subset of microRNAs was preferentially released in exosomes others were selectively retained in the cells. Moreover, exposure of MIN6B1 cells to inflammatory cytokines changed the release of several microRNAs. The dynamics of microRNA secretion and their potential transfer to recipient cells were next investigated. As a proof-of-concept, we demonstrate that if cel-miR-238, a C. Elegans microRNA not present in mammalian cells, is expressed in MIN6B1 cells a fraction of it is released in exosomes and is transferred to recipient beta-cells. Furthermore, incubation of untreated MIN6B1 or mice islet cells in the presence of microRNA-containing exosomes isolated from the culture media of cytokine-treated MIN6B1 cells triggers apoptosis of recipient cells. In contrast, exosomes originating from cells not exposed to cytokines have no impact on cell survival. Apoptosis induced by exosomes produced by cytokine-treated cells was prevented by down-regulation of the microRNA-mediating silencing protein Ago2 in recipient cells, suggesting that the effect is mediated by the non-coding RNAs. CONCLUSIONS: Taken together, our results suggest that beta-cells secrete microRNAs that can be transferred to neighboring beta-cells. Exposure of donor cells to pathophysiological conditions commonly associated with diabetes modifies the release of microRNAs and affects survival of recipient beta-cells. Our results support the concept that exosomal microRNAs transfer constitutes a novel cell-to-cell communication mechanism regulating the activity of pancreatic beta-cells.

Seleção de clones de figueira cv. roxo de Valinhos formados por gemas irradiadas

A figueira (Ficus carica L.), pertencente à família das Moráceas, constitui-se numa das mais importantes frutíferas cultivadas, elevando o Brasil à condição de décimo maior produtor e exportador de figos do mundo. Porém, a ficicultura apresenta alguns problemas fitossanitários, além de, no Brasil, estar toda implantada com uma única cultivar, a Roxo de Valinhos, que produz frutos sem sementes, inviabilizando o melhoramento convencional. Nesse sentido, o melhoramento genético, com o uso de mutagênicos, passa a ser uma linha de pesquisa altamente importante, podendo contribuir enormemente para o desenvolvimento da cultura. Diante disto, o objetivo do presente trabalho foi selecionar mutantes em plantas de figueira formadas por estacas irradiadas com raios gama, a fim de aumentar sua variabilidade genética com relação ao desenvolvimento vegetativo e reprodutivo. Utilizaram-se plantas formadas por estacas originadas de gemas da cultivar Roxo de Valinhos irradiadas com raios gama, no irradiador tipo Gamma a 0,10 m do ápice, na dose de Gy com taxa de dose de 238 Gy/h. O experimento constou de 450 tratamentos, sendo cada planta formada considerada um tratamento, numerando-as sequencialmente de 1 a 450 e cultivadas em espaçamento de 2,5 x 1,5 m.. As avaliações foram realizadas a partir das características tanto das folhas quanto dos frutos, bem como da incidência das principais pragas e doenças da cultura nestas plantas. Da análise dos dados, conclui-se que há variabilidade genética entre os tratamentos e que algumas plantas são prováveis mutantes, mostrando-se assim com potencial para posteriores estudos, devendo ser testadas em plantios comerciais.

Predição simultânea dos efeitos de marcadores moleculares e seleção genômica ampla em cajueiro

A seleção genômica ampla (genome wide selection - GWS) foi proposta como uma forma de aumentar a eficiência e acelerar o melhoramento genético, enfatizando a predição simultânea dos efeitos genéticos de grande número de marcadores genéticos de DNA dispersos em todo o genoma de um organismo, de forma a capturar os efeitos de todos os locos e explicar a variação genética de um caráter quantitativo. Objetivou-se com o presente trabalho aplicar o princípio da GWS no melhoramento do cajueiro, estimando simultaneamente os efeitos de 238 marcadores avaliados em 74 indivíduos de uma família de irmãos completos, visando a explicar grande porcentagem da variação genotípica total do caráter peso da amêndoa e a aumentar a eficiência do melhoramento do cajueiro. Verificou-se que a capacidade preditiva e a acurácia são praticamente maximizadas na análise com 70 marcadores de maiores efeitos. O aumento do número de marcadores não aumenta linearmente a acurácia da GWS pelo método RR-BLUP. Os 70 marcadores de maiores efeitos capturam 74% da variação genotípica total e propiciam alta acurácia seletiva (86%) da seleção para o peso de amêndoas, enquanto os cinco marcadores de maiores efeitos capturam apenas 19% da variação genotípica total e propiciam acurácia seletiva de apenas 44%. Assim, a seleção assistida (MAS), baseada em poucos (cinco) marcadores de efeitos significativos, propicia eficiência muito inferior à GWS. Os valores genéticos genômicos preditos na população de validação cruzada aproximam-se bem dos valores fenotípicos observados, com correlação de 0,79. A estimação simultânea dos efeitos dos marcadores, segundo o conceito da GWS, é uma alternativa interessante, visando a aumentar a eficiência do melhoramento do cajueiro.

Reproducibility of Vertebral Fracture Assessment Readings From Dual-energy X-ray Absorptiometry in Both a Population-based and Clinical Cohort: Cohen's and Uniform Kappa.

Vertebral fracture assessments (VFAs) using dual-energy X-ray absorptiometry increase vertebral fracture detection in clinical practice and are highly reproducible. Measures of reproducibility are dependent on the frequency and distribution of the event. The aim of this study was to compare 2 reproducibility measures, reliability and agreement, in VFA readings in both a population-based and a clinical cohort. We measured agreement and reliability by uniform kappa and Cohen's kappa for vertebral reading and fracture identification: 360 VFAs from a population-based cohort and 85 from a clinical cohort. In the population-based cohort, 12% of vertebrae were unreadable. Vertebral fracture prevalence ranged from 3% to 4%. Inter-reader and intrareader reliability with Cohen's kappa was fair to good (0.35-0.71 and 0.36-0.74, respectively), with good inter-reader and intrareader agreement by uniform kappa (0.74-0.98 and 0.76-0.99, respectively). In the clinical cohort, 15% of vertebrae were unreadable, and vertebral fracture prevalence ranged from 7.6% to 8.1%. Inter-reader reliability was moderate to good (0.43-0.71), and the agreement was good (0.68-0.91). In clinical situations, the levels of reproducibility measured by the 2 kappa statistics are concordant, so that either could be used to measure agreement and reliability. However, if events are rare, as in a population-based cohort, we recommend evaluating reproducibility using the uniform kappa, as Cohen's kappa may be less accurate.

Status epilepticus: impact of therapeutic coma on outcome.

OBJECTIVES: Therapeutic coma is advocated in guidelines for management of refractory status epilepticus; this is, however, based on weak evidence. We here address the specific impact of therapeutic coma on status epilepticus outcome. DESIGN: Retrospective assessment of a prospectively collected cohort. SETTING: Academic hospital. PATIENTS: Consecutive adults with incident status epilepticus lasting greater than or equal to 30 minutes, admitted between 2006 and 2013. MEASUREMENTS AND MAIN RESULTS: We recorded prospectively demographics, clinical status epilepticus features, treatment, and outcome at discharge and retrospectively medical comorbidities, hospital stay, and infectious complications. Associations between potential predictors and clinical outcome were analyzed using multinomial logistic regressions. Of 467 patients with incident status epilepticus, 238 returned to baseline (51.1%), 162 had new disability (34.6%), and 67 died (14.3%); 50 subjects (10.7%) were managed with therapeutic coma. Therapeutic coma was associated with poorer outcome in the whole cohort (relative risk ratio for new disability, 6.86; 95% CI, 2.84-16.56; for mortality, 9.10; 95% CI, 3.17-26.16); the effect was more important in patients with complex partial compared with generalized convulsive or nonconvulsive status epilepticus in coma. Prevalence of infections was higher (odds ratio, 3.81; 95% CI, 1.66-8.75), and median hospital stay in patients discharged alive was longer (16 d [range, 2-240 d] vs 9 d [range, 1-57 d]; p < 0.001) in subjects managed with therapeutic coma. CONCLUSIONS: This study provides class III evidence that therapeutic coma is associated with poorer outcome after status epilepticus; furthermore, it portends higher infection rates and longer hospitalizations. These data suggest caution in the straightforward use of this approach, especially in patients with complex partial status epilepticus.

Reação de aceroleira (Malpighia emarginata D.C.) à Meloidogyne enterolobii

O Brasil é o maior produtor, consumidor e exportador de acerola (Malpighia emarginata D.C.) do mundo. Por conter altos teores de vitamina C, tornou-se uma fruta altamente requisitada no mercado mundial para o preparo de sucos e no consumo in natura. Nos últimos anos, as lavouras desta fruta vêm apresentando um decréscimo nas produções em razäo da ocorrência de nematoides de galhas (Meloidogyne spp.), um dos principais problemas que afetam a cultura. O objetivo deste trabalho foi avaliar a resistência de genótipos de aceroleira frente à Meloidogyne enterolobii. O experimento foi conduzido em casa de vegetação do Departamento de Proteção Vegetal, da Faculdade de Ciências Agronômicas/UNESP - Câmpus de Botucatu (SP). Foram utilizados cinco clones: Cereja-Brs-236; Fruta Cor- Brs-238; Roxinha-Brs-237; Mirandópolis; Japi, e três variedades: Okinawa; Olivier e Waldy-CATI. Cada planta foi inoculada com 2.500 ovos e eventuais juvenis de segundo estádio (Pi) de M. enterolobii. Após 60 dias, a parte área de cada planta foi descartada, e o sistema radicular lavado, submetido à coloração com floxina-B e examinado para a obtenção dos índices de galhas (IG) e massa de ovos (IMO), e processados pelo método de trituração em liquidificador, peneiramento e centrifugação com sacarose para a obtenção do número total de ovos (Pf), que foi utilizado para o cálculo do fator de reprodução (Pf/Pi). Todos os clones e as variedades foram considerados suscetíveis à Meloidogyne enterolobii apresentando os fatores de reprodução variando de 4,1 a 18,3.

M.A.T.

La memòria del projecte que us presentem, tracta el procés de realització del Treball Final de Grau, M.A.T.. Aquest és un projecte audiovisual i multimèdia creat per set alumnes de Comunicació Audiovisual de la Universitat de Vic. El grup encarregat de la part audiovisual, format per la Dámaris Alonso, en Guillem Bover, en Raúl Caso i la Irina Mora, ha creat un curtmetratge lineal. M.A.T. és un projecte que sorgeix per impulsar el gènere del thriller dins de les produccions catalanes. A més a més, la nostra idea ha consistit en generar un producte que es pogués distribuir a través de plataformes interactives. Per aquest motiu, el projecte es divideix en un curtmetratge audiovisual i un joc interactiu inclòs a la pàgina oficial. Els integrants del grup ens vam encarregar de desenvolupar la peça audiovisual, la qual narra una història de venjança i traïció en un entorn de la màfia actual del nostre país. Una de les principals característiques d’aquest projecte és el tractament del temps i del ritme de la narració. És a dir, hem creat una trama audiovisual que s’estén durant quinze anys i hem generat el suspens que s’escau segons els paràmetres del gènere cinematogràfic de la peça. Un factor clau d’aquest projecte és una partida d’escacs, la qual juga un paper molt important, no tan sols en la història del curtmetratge, sinó perquè és la forma d’interacció que fa servir l’usuari a la pàgina web.

Radiotherapy for GIST progressing during or after tyrosine kinase inhibitor therapy: A prospective study.

PURPOSE: Gastrointestinal stromal tumor (GIST) has been considered radiation-resistant, and radiotherapy is recommended only for palliation of bone metastases in current treatment guidelines. No registered prospective trial has evaluated GIST responsiveness to radiotherapy. PATIENTS AND METHODS: Patients with GIST progressing at intra-abdominal sites or the liver were entered to this prospective Phase II multicenter study (identifier NCT00515931). Metastases were treated with external beam radiotherapy using either conformal 3D planning or intensity modulated radiotherapy and conventional fractionation to a cumulative planning target volume dose of approximately 40Gy. Systemic therapy was maintained unaltered during the study. RESULTS: Of the 25 patients entered, 19 were on concomitant tyrosine kinase inhibitor therapy, most often imatinib. Two (8%) patients achieved partial remission, 20 (80%) had stable target lesion size for ⩾3months after radiotherapy with a median duration of stabilization of 16months, and 3 (12%) progressed. The median time to radiotherapy target lesion progression was 4-fold longer than the median time to GIST progression at any site (16 versus 4months). Radiotherapy was generally well tolerated. CONCLUSIONS: Responses to radiotherapy were infrequent, but most patients had durable stabilization of the target lesions. GIST patients with soft tissue metastases benefit frequently from radiotherapy.

Eficacia del kinesiotape como terapia complementaria en niños que presentan espasticidad.

¿Se obtiene una mejora de resultados añadiendo Kinesiotape a sujetos que presentan PCI espástica comparado con un tratamiento convencional de fisioterapia? Objetivos: Evaluar la efectividad del Kinesiotape en cuanto a disminución de espasticidad y tono muscular, mejora de calidad de vida y aumento del rango articular. Material y métodos: se buscará una muestra de 238 sujetos con PCI espástica entre 5 y 20 años y que presenten una sinergia flexora en extremidad superior. Como criterios de exclusión se consideran: cualquier contraindicación del Kinesiotape, intervención quirúrgica de extremidad superior, consumo de neurotoxinas, bloqueantes nerviosos con agentes neurolíticos, relajantes musculares y pacientes que hayan sido tratados mediante Toxina Botulínica en extremidad superior. Los sujetos serán aleatoriamente distribuidos en dos grupos; un grupo control (n = 119) el cual realiza tratamiento específico de Bobath y un grupo experimental (n = 119) el cual realiza el tratamiento específico de Bobath añadiendo el Kinesiotape en la musculatura del bíceps braquial. Esta intervención durará ocho semanas y se reevaluarán los resultados en un periodo de seis meses y a los doce meses. Para ello se utilizaran las siguientes escalas: para la espasticidad se utilizarán las escalas de Tardieu - Held y la de Aswhorth modificada. Para medir la independencia del niño se emplearán la escala Pediatric Evaluation of Disability Inventory (PEDI), para conocer el rango articular se utilizarán un goniómetro de brazos y para determinar la actividad eléctrica del tono muscular la electromiografía (EMG).

Conocimiento y satisfacción de la aprobación de la prescripción enfermera (Ley 28/2009) en enfermeras de formación máster y especialidad de la Universidad de Barcelona

Objetivo: Identificar el nivel de conocimiento, satisfacción y capacitación de las enfermeras con relación a la prescripción enfermera tras la aprobación de la Ley 28/2009. Método: Estudio observacional descriptivo transversal realizado en la Escuela de Enfermería de la Universidad de Barcelona. Los sujetos a estudio fueron 485 enfermeras en formación máster o especialización y docentes de dicha escuela. Las variables principales de estudio fueron: satisfacción, conocimiento e información de la aprobación de la Ley, conocimiento de los elementos autorizados a prescribir, percepción de capacidad prescriptora de las enfermeras y opinión de la necesidad formativa e informativa respecto a la Ley. Como secundarias: años de experiencia profesional, ámbito laboral, curso formativo y sociodemográficas. La recogida de datos se realizó entre los meses de marzo y abril de 2010 mediante un cuestionario autoaplicable ad hoc de 29 ítems, probado previamente. Resultados: Participaron 345 enfermeras, el 87,2% (301) alumnas en formación máster. El 72,3% (238) de las enfermeras conocían la aprobación de la ley, pero un 95,9% (303) indicó tener insuficiente información. La satisfacción media con la aprobación de la Ley se situó en 6,24 ± 2,08. El 57,6% (189) de las enfermeras opinaron estar capacitadas para prescribir productos sanitarios y un 41,3% (134), para prescribir fármacos. Conclusiones: Hay satisfacción con la aprobación de la Ley 28/2009. Actualmente muchas enfermeras prescriben productos farmacológicos y sanitarios de forma autónoma.

Animal modelling for inherited central vision loss.

Disease-causing variants of a large number of genes trigger inherited retinal degeneration leading to photoreceptor loss. Because cones are essential for daylight and central vision such as reading, mobility, and face recognition, this review focuses on a variety of animal models for cone diseases. The pertinence of using these models to reveal genotype/phenotype correlations and to evaluate new therapeutic strategies is discussed. Interestingly, several large animal models recapitulate human diseases and can serve as a strong base from which to study the biology of disease and to assess the scale-up of new therapies. Examples of innovative approaches will be presented such as lentiviral-based transgenesis in pigs and adeno-associated virus (AAV)-gene transfer into the monkey eye to investigate the neural circuitry plasticity of the visual system. The models reported herein permit the exploration of common mechanisms that exist between different species and the identification and highlighting of pathways that may be specific to primates, including humans.

Acute and Chronic Altitude-Induced Cognitive Dysfunction in Children and Adolescents.

OBJECTIVE: To assess whether exposure to high altitude induces cognitive dysfunction in young healthy European children and adolescents during acute, short-term exposure to an altitude of 3450 m and in an age-matched European population permanently living at this altitude. STUDY DESIGN: We tested executive function (inhibition, shifting, and working memory), memory (verbal, short-term visuospatial, and verbal episodic memory), and speed processing ability in: (1) 48 healthy nonacclimatized European children and adolescents, 24 hours after arrival at high altitude and 3 months after return to low altitude; (2) 21 matched European subjects permanently living at high altitude; and (3) a matched control group tested twice at low altitude. RESULTS: Short-term hypoxia significantly impaired all but 2 (visuospatial memory and processing speed) of the neuropsychological abilities that were tested. These impairments were even more severe in the children permanently living at high altitude. Three months after return to low altitude, the neuropsychological performances significantly improved and were comparable with those observed in the control group tested only at low altitude. CONCLUSIONS: Acute short-term exposure to an altitude at which major tourist destinations are located induces marked executive and memory deficits in healthy children. These deficits are equally marked or more severe in children permanently living at high altitude and are expected to impair their learning abilities.

Mammographic density in asymptomatic menopausal women: correlation with clinical and sonographic findings

OBJECTIVE: To evaluate mammographic breast density in asymptomatic menopausal women in correlation with clinical and sonographic findings. MATERIALS AND METHODS: Mammograms and clinical and sonographic findings of 238 asymptomatic patients were retrospectively reviewed in the period from February/2022 to June/2006. The following variables were analyzed: mammographic density patterns, sonographic findings, patients' age, parity, body mass index and use of hormone replacement therapy. RESULTS: Age, parity and body mass index showed a negative correlation with breast density pattern, while use of hormone replacement therapy showed a positive correlation. Supplementary breast ultrasonography was performed in 103 (43.2%) patients. Alterations which could not be visualized at mammography were found in 34 (33%) of them, most frequently in women with breast density patterns 3 and 4. CONCLUSION: The authors concluded that breast density patterns were influenced by age, parity, body mass index and time of hormone replacement therapy. Despite not having found any malignant abnormality in the studied cases, the authors have observed a predominance of benign sonographic abnormalities in women with high breast density patterns and without mammographic abnormalities, proving the relevance of supplementary ultrasonography to identify breast lesions in such patients.

Similar Hemoglobin Mass Response in Hypobaric and Normobaric Hypoxia in Athletes

PURPOSE: To compare hemoglobin mass (Hbmass) changes during an 18-d live high-train low (LHTL) altitude training camp in normobaric hypoxia (NH) and hypobaric hypoxia (HH). METHODS: Twenty-eight well-trained male triathletes were split into three groups (NH: n = 10, HH: n = 11, control [CON]: n = 7) and participated in an 18-d LHTL camp. NH and HH slept at 2250 m, whereas CON slept, and all groups trained at altitudes <1200 m. Hbmass was measured in duplicate with the optimized carbon monoxide rebreathing method before (pre-), immediately after (post-) (hypoxic dose: 316 vs 238 h for HH and NH), and at day 13 in HH (230 h, hypoxic dose matched to 18-d NH). Running (3-km run) and cycling (incremental cycling test) performances were measured pre and post. RESULTS: Hbmass increased similar in HH (+4.4%, P < 0.001 at day 13; +4.5%, P < 0.001 at day 18) and NH (+4.1%, P < 0.001) compared with CON (+1.9%, P = 0.08). There was a wide variability in individual Hbmass responses in HH (-0.1% to +10.6%) and NH (-1.4% to +7.7%). Postrunning time decreased in HH (-3.9%, P < 0.001), NH (-3.3%, P < 0.001), and CON (-2.1%, P = 0.03), whereas cycling performance changed nonsignificantly in HH and NH (+2.4%, P > 0.08) and remained unchanged in CON (+0.2%, P = 0.89). CONCLUSION: HH and NH evoked similar Hbmass increases for the same hypoxic dose and after 18-d LHTL. The wide variability in individual Hbmass responses in HH and NH emphasizes the importance of individual Hbmass evaluation of altitude training.