The emergence of <i>Pax7</i>-expressing muscle stem cells during vertebrate head muscle development

Pax7 expressing muscle stem cells accompany all skeletal muscles in the body and in healthy individuals, efficiently repair muscle after injury. Currently, the in vitro manipulation and culture of these cells is still in its infancy, yet muscle stem cells may be the most promising route towards the therapy of muscle diseases such as muscular dystrophies.It is often overlooked that muscular dystrophies affect head and body skeletal muscle differently. Moreover, these muscles develop differently. Specifically, head muscle and its stem cells develop from the non-somitic head mesoderm which also has cardiac competence. To which extent head muscle stem cells retain properties of the early head mesoderm and might even be able to switch between a skeletal muscle and cardiac fate is not known. This is due to the fact that the timing and mechanisms underlying head muscle stem cell development are still obscure. Consequently, it is not clear at which time point one should compare the properties of head mesodermal cells and head muscle stem cells.To shed light on this, we traced the emergence of head muscle stem cells in the key vertebrate models for myogenesis, chicken, mouse, frog and zebrafish, using Pax7 as key marker. Our study reveals a common theme of head muscle stem cell development that is quite different from the trunk. Unlike trunk muscle stem cells, head muscle stem cells do not have a previous history of Pax7 expression, instead Pax7 expression emerges de-novo. The cells develop late, and well after the head mesoderm has committed to myogenesis. We propose that this unique mechanism of muscle stem cell development is a legacy of the evolutionary history of the chordate head mesoderm

Utilização do Soro de Queijo na Potencialização da Produção de Etanol a Partir de Três Cultivares de Arroz

Rice cultivation has been the subject of research that involves its production chain, including the study of the use of its residues, from the cultivation to its processing. The use of whey has also been well studied, as this by-product causes great environmental impact if not treated with due caution. In this context, the objective of this study was to evaluate the use of whey in ethanol production potentiation from three rice varieties, with ethanol as final product. The residue of ethanol production was an alternative for industries and dairy products, since the means in which the whey was added had a value equal to and above 28.11% of protein. Analyzing the kinetics of the conversion of glucose into ethanol, the peak was reached at approximately 19 hours, ie the best treatment, where it was able to reach the highest concentration of ethanol. The treatment that obtained the best result was the serum of 50% in the Serra Dourada variety, where its production was 78.52 g / L of ethanol. The averages of both did not differ statistically, obtaining at the 5% level of significance by the Tukey test. The results showed that there is a technical feasibility in the incorporation of whey in the conventional process of ethanol production from the rice, optimizing the reuse of this co-product of the dairy industry.A cultura do arroz vem sendo alvo de pesquisas envolvendo sua cadeia produtiva, incluindo o estudo de aproveitamentos de seus resíduos, desde a lavoura até o seu beneficiamento. O aproveitamento do soro de queijo também tem sido bastante estudado, pois esse derivado causa grande impacto ambiental se não for tratado devidamente. Neste contexto o presente trabalho objetivou avaliar a utilização do soro de queijo na potencialização da produção de etanol a partir de três variedades de arroz tendo como produto final o etanol. O co-produto da produção de etanol mostrou-se uma alternativa para alimentação animal, apresentaram valor igual e superior a 28,11% de proteína. Analisando a cinética da conversão da glicose em etanol o ponto máximo foi atingido aproximadamente com 19 horas, quando se conseguiu atingir a maior concentração de etanol. O tratamento que obteve melhor resultado foi o mosto contendo 50% de soro para variedade Serra Dourada, onde a sua produção foi de 78,52 g/L de etanol. Os resultados obtidos mostraram, portanto, que há viabilidade técnica na incorporação do soro de queijo no processo convencional de produção de etanol a partir do arroz, otimizando-se o reaproveitamento deste co-produto da indústria de laticínio

The elderly’s look regarding the access to a basic health unit in Porto Alegre - RS

Com o crescente envelhecimento da população brasileira, fica evidente a necessidade de maior preparo e planejamento dos serviços de saúde para atender os idosos. Objetivos: Analisar como se dá o acesso da população idosa ao serviço de saúde em uma Unidade Básica de Saúde de Porto Alegre, pois nunca se obteve dados fundamentados para avaliar a assistência aos idosos nessa unidade de saúde. Materiais e métodos: Este estudo foi elaborado de uma forma descritiva e transversal, através de entrevistas semi-estruturadas contendo quinze questões abertas e fechadas baseadas no PCA-Tool como instrumento, com uma amostra de 61 pacientes idosos. Resultados: A Unidade de Saúde SESC foi lembrada pela grande maioria da população idosa como serviço de referência para primeiras necessidades em saúde. As maiores dificuldades de acesso se deram em relação às características geográficas do território onde a Unidade está inserida. É necessária uma reflexão sobre que medidas devem ser tomadas para solucionar este problema. O acesso dos idosos às modalidades de consulta médica, odontológica e de enfermagem foi considerado facilitado. Porém, a percepção desses usuários quanto às estratégias de priorização no atendimento não se apresentou tão evidente. Conclusão: A partir da opinião dessa população, apesar de algumas dificuldades assinaladas, o acesso ao serviço de saúde foi considerado facilitado para os idosos nessa Unidade de Saúde.Objective: The aim of this study was to discribe how to give the elderly access to health services in a Basic Health Unit in Porto Alegre since it substantiated data was never available to evaluate the care for the elderly population in this health unit. Materials and methods: This study was conducted with a descriptive and cross-sectional design, applying semi-structured interviews with fifteen open and closed questions based on PCA-Tool as an instrument in a sample of 61 elderly patients. Results: The main results show that the Health Unit SESC was remembered by the vast majority of the elderly population as a reference service for basic needs in health. The greatest difficulties of access were related to the characteristics of an area where the Unit is located. It is necessary to consider what measures should be taken to solve this problem. The access of older methods of medical, dental and nursing was considered easier. However, the perception of users about the strategies for prioritization in attendance did not appear so clear. Conclusion: It was concluded from the review of this population that, despite some difficulties pointed out, access to health services was considered easier for the elderly in this Health Unit

Produção de proteinas recombinantes utilizando Escherichia coli em cultivos em alta densidade celular

Orientador: Maria Helena Andrade SantanaTese (doutorado) - Universidade Estadual de Campinas, Faculdade de Engenharia de QuimicaResumo: Neste trabalho foi possível estudar o cultivo em altas densidades celulares de E. coli recombinante em bioreatores identificando importantes parâmetros cinéticos para as fases de crescimento celular e indução da proteína recombinante. Foram estudadas diversas estratégias de cultivo para obtenção de elevadas densidades celulares, no intuito de desenvolver' uma tecnologia de fermentação robusta que permitisse: Elevados níveis de biomassa em peso seco de células por litro; determinar as condições nutricionais que permitem expressão da proteína de fusão da ordem de 20% ou mais em fermentações com alta concentração de biomassa e determinar a melhor estratégia de fermentação para atingir elevadas concentrações celulares, entre as quais, batelada-alimentada com cortes, batelada-alimentada cíclica em dois estágios e fermentações com reciclo e microfiltração. A cepa utilizada neste estudo foi a E. coli N-4830-1 transformada com o plasmídeo pHis. As fermentações foram realizadas em bioreatores de bancada de capacidade útil de 4,0 L e 12,0 L (NBS e Inceltech). Foram medidas as concentrações de biomassa, glicose, acetato e proteína recombinante. Os estudos cinéticos das diferentes estratégias mostraram incrementos em biomassa e em proteína de fusão, sendo que o melhor resultado foi obtido para fermentações com reciclo total de células, microfiltração do meio e alimentação exponencial de nutrientes, as quais obtiveram produtividade celular igual a PR = 6,437 g/L/h. A concentração final em peso seco de células foi 173,8 g/L, com porcentagem de expressão da proteína de fusão em 17,8%. Os incrementos obtidos em biomassa e em proteína de fusão, em relação à batelada-alimentada tradicional, foram respectivamente 341 % e 328 %. A técnica mostrou-se adequada para remoção de inibidores no meio de cultura permitindo maior produtividade e, conseqüentemente, maior crescimento celular, sendo até 3,5 vezes superiores aos cultivos em batelada-alimentada tradicionalAbstract: This thesis studied submerged high cell-density cultures (HCDC) of recombinant Escherichia coli identifying important kinetic parameters on growth and production phases. Several strategies to obtain HCDC were studied aiming to develop a robust fermentation process that allowed: High cell dry weight per liter; to determine the nutritional conditions to express the fusion protein in levels up to 20% of total protein, and to determine the best fermentation technique to reach dry biomass concentration higher that 100,0 g/L. The strain used in this work was E. colí N-4830-1 transformed with the plasmid pHis. The fermentations were ran out in 4.0 L and 12.0 L bench-top bioreactors (NBS e Inceltech). Total protein concentration, recombinant protein concentration, cell concentration, glucose concentration and acetate concentration were analyzed. The kinetics studies demonstrated that biomass and fusion protein were increased, however the best results were performed on a continuous fermentation with microfiltration, total cell recycle and exponencial feeding. The cell productivity was PR = 6,437 glUh, the final cell dry weight was 173,8 g/L and the percentage of fusion protein was 17,8 % of total protein. The biomass and fusion protein increase obtained were, respectively, 341 % and 328 %, comparing with traditional fed-batch fermentation. This technique showed appropriated to remove inhibitors of the broth allowing higher cell concentration and, consequently, higher productivity. In this type of culture strategy, the final cell concentration was 3.5 fold than one obtained in traditional fed-batchDoutoradoDesenvolvimento de Processos BiotecnologicosDoutor em Engenharia Químic

Scalar and Pseudo-Scalar Higgs Production at Hadron Colliders

The evaluation of the NNLO QCD corrections to the production of a scalar and a pseudo-scalar Higgs boson is described.Comment: 5+2 pages, 4 figures (9 ps-files). Talk given at RADCOR/Loops and Legs 2002, September 8-13, 2002, Kloster Banz, German

Associations Between Declining Antibiotic Use in Primary Care in Scotland and Hospitalisation with Infection and Patient Satisfaction:Longitudinal Population Study

BACKGROUND: Reducing antibiotic use is central to antimicrobial stewardship, but may have unintended consequences. OBJECTIVES: To examine associations between size of decline in antibiotic prescriptions in general practices and (i) rate of hospitalization for infection and (ii) patient satisfaction. METHODS: Routine data analysis for all general practices in Scotland, quarter one 2012 (Q1 2012) to quarter one 2018 (Q1 2018). Practices were grouped into quartiles of rate of change in prescribing and changes in rates of hospitalization were compared across groups. For satisfaction analysis, associations between practice-level patient satisfaction in 2017–18 (Scottish Health and Care Experience Survey) and prior change in antibiotic prescription were examined. RESULTS: Antibiotic prescriptions overall fell from 194.1 prescriptions/1000 patients in Q1 2012 to 165.3 in Q1 2018 (14.9% reduction). The first quartile of practices had a non-significant increase in prescriptions [change per quarter = 0.22 (95% CI −0.42 to 0.86) prescriptions/1000 patients], compared with large reductions in the other three groups, largest in quartile four: −2.95 (95% CI −3.66 to −2.24) prescriptions/1000 patients/quarter (29.7% reduction overall). In all quartiles, hospitalizations with infection increased. The increase was smallest in quartile four (the biggest reduction in prescriptions) and highest in quartile one (no significant change in prescriptions): 2.18 (95% CI 1.18 to 3.19) versus 3.68 (95% CI 2.64 to 4.73) admissions/100 000 patients/quarter, respectively [difference = − 1.50 (95% CI −2.91 to −0.10)]. There was no statistically significant association between change in antibiotic prescriptions and patient satisfaction. CONCLUSIONS: Very large reductions in antibiotic prescriptions in Scottish general practices have not been associated with increases in hospitalization with infection or changes in patient satisfaction

4-aminopyridyl-based lead compounds targeting CYP51 prevent spontaneous parasite relapse in a chronic model and improve cardiac pathology in an acute model of Trypanosoma cruzi infection.

BackgroundChagas disease, caused by the protozoan Trypanosoma cruzi, is the leading cause of heart failure in Latin America. The clinical treatment of Chagas disease is limited to two 60 year-old drugs, nifurtimox and benznidazole, that have variable efficacy against different strains of the parasite and may lead to severe side effects. CYP51 is an enzyme in the sterol biosynthesis pathway that has been exploited for the development of therapeutics for fungal and parasitic infections. In a target-based drug discovery program guided by x-ray crystallography, we identified the 4-aminopyridyl-based series of CYP51 inhibitors as being efficacious versus T.cruzi in vitro; two of the most potent leads, 9 and 12, have now been evaluated for toxicity and efficacy in mice.Methodology/principal findingsBoth acute and chronic animal models infected with wild type or transgenic T. cruzi strains were evaluated. There was no evidence of toxicity in the 28-day dosing study of uninfected animals, as judged by the monitoring of multiple serum and histological parameters. In two acute models of Chagas disease, 9 and 12 drastically reduced parasitemia, increased survival of mice, and prevented liver and heart injury. None of the compounds produced long term sterile cure. In the less severe acute model using the transgenic CL-Brenner strain of T.cruzi, parasitemia relapsed upon drug withdrawal. In the chronic model, parasitemia fell to a background level and, as evidenced by the bioluminescence detection of T. cruzi expressing the red-shifted luciferase marker, mice remained negative for 4 weeks after drug withdrawal. Two immunosuppression cycles with cyclophosphamide were required to re-activate the parasites. Although no sterile cure was achieved, the suppression of parasitemia in acutely infected mice resulted in drastically reduced inflammation in the heart.Conclusions/significanceThe positive outcomes achieved in the absence of sterile cure suggest that the target product profile in anti-Chagasic drug discovery should be revised in favor of safe re-administration of the medication during the lifespan of a Chagas disease patient. A medication that reduces parasite burden may halt or slow progression of cardiomyopathy and therefore improve both life expectancy and quality of life

Different packing motifs of isomeric (E)-N1-(halophenylmethylidene)-N-methyl-2-(thiophen-2-yl)-acetohydrazides controlled by C—HO interactions

We thank the EPSRC National Crystallography Service (University of Southampton) for the X-ray data collectionsPeer reviewedPublisher PD

Liquid-liquid equilibria in aqueous two-phase ethanol/salt systems at different temperatures and their application to anthocyanins extraction

Abstract Aqueous two-phase systems (ATPS) are formed by mixtures of chemical species that, under certain conditions, separate into two immiscible phases, with water as the major component in global terms and, at lower concentrations, polymers, salts, ionic liquids and alcohols, depending on the system type. Different parameters influence ATPS equilibrium such as temperature, alcohol carbon chain size, salt type, pH, etc. The present work aimed to obtain the binodal curves for ATPS made up of ethanol, sodium citrate/ammonium sulfate and water at different temperatures (15, 25 and 35 °C) as well as to evaluate the potential of the ethanol/ammonium sulfate system in anthocyanins partition. The results showed that the increase in temperature did not alter the biphasic region in the temperature range investigated, not influencing the formation of phases. Nonlinear equations were satisfactorily fitted to binodal curves data, except for sodium citrate-containing ATPS at 15 °C. Higher concentration of the overall mixture resulted in longer tie-line of ammonium sulphate-containing ATPS. Higher values of partition coefficient and recovery yield of Syzygium cumini fruit anthocyanins were obtained at longer tie-lines