Impacto do pH final na maciez do músculo Longissimus lumborum de animais zebuínos: mudanças estruturais de proteínas da carne crua e cozida durante a maturação

O objetivo do presente trabalho foi determinar a força de cisalhamento Warner-Bratzler do músculo Longissimus lumborum de animais zebuínos machos inteiros (Bos indicus) durante o período de maturação, nas faixas de pH final (pHf 48 horas post mortem) normal (pH entre 5,5 e 5,8) e anormal (pH entre 5,81 e 6,19) e temperaturas internas de cozimento. Concomitante com a avaliação de força de cisalhamento, foram avaliadas também a degradação da desmina e troponina T, o comprimento do sarcômero, o teor de colágeno total e solúvel, as temperaturas máximas de desnaturação das proteínas e a morfologia geral de agregação das fibras do músculo no cozimento. A degradação da desmina e troponina T foi maior no pHf normal, aparecendo produtos de degradação a partir do dia 7 nessa faixa de pHf. Não houve diferenças nos valores de comprimento do sarcômero, descartando-se assim, a contribuição desse parâmetro sobre a temperatura máxima de desnaturação (Tmáx) das proteínas, determinada utilizando calorímetro exploratório diferencial (DSC). Similarmente, não foram encontradas diferenças para os teores de colágeno total e solúvel, e os valores de colágeno total foram baixos, sugerindo que sua contribuição na segunda transição térmica e nos valores de força de cisalhamento foi mínima. As Tmáx1 e Tmáx2, correspondentes à desnaturação da meromiosina leve e pesada, respectivamente, foram menores no pHf normal, mas o efeito foi maior para a Tmáx2. A Tmáx3 da actina e titina aumentou até 14 dias post mortem na faixa de pHf normal, e posteriormente diminuiu significativamente após 21 dias, sugerindo possível degradação dessas proteínas nesse período de dias. Não foram encontradas diferenças nos valores de Tmáx no pHf anormal, em todos os dias post mortem, o que sugere a contribuição de um possível mecanismo de proteção que estabiliza as miofibrilas no aquecimento. Houve maior agregação das fibras do músculo no pHf normal nas temperaturas internas de cozimento de 65 e 80°C, provavelmente devido à maior desnaturação térmica das miofibrilas. Os valores de força de cisalhamento foram maiores com o aumento da temperatura interna de cozimento, devido ao aumento da desnaturação térmica das miofibrilas do músculo. Independente da temperatura interna de cozimento, os valores de força de cisalhamento foram altos em quase todos os dias post mortem para ambas as faixas de pHf, o que sugere a necessidade de utilizar métodos físicos ou químicos para aumentar a maciez do músculo Longissimus lumborum de animais zebuínos.

Involvement of loop 5 lysine residues and the N-terminal β-hairpin of the ribotoxin hirsutellin A on its insecticidal activity

Ribotoxins are cytotoxic members of the family of fungal extracellular ribonucleases best represented by RNase T1. They share a high degree of sequence identity and a common structural fold, including the geometric arrangement of their active sites. However, ribotoxins are larger,with a well-defined N-terminal β-hairpin, and display longer and positively charged unstructured loops. These structural differences account for their cytotoxic properties.Unexpectedly, the discovery of hirsutellin A (HtA), a ribotoxin produced by the invertebrate pathogen Hirsutella thompsonii, showed how it was possible to accommodate these features into a shorter amino acid sequence. Examination of HtA N-terminal β-hairpin reveals differences in terms of length, charge, and spatial distribution. Consequently,four different HtA mutants were prepared and characterized. One of them was the result of deleting this hairpin [Δ(8-15)] while the other three affected single Lys residues in its close spatial proximity (K115E, K118E, and K123E). The results obtained support the general conclusion that HtA active site would show a high degree of plasticity,being able to accommodate electrostatic and structural changes not suitable for the other previously known larger ribotoxins, as the variants described here only presented small differences in terms of ribonucleolytic activity and cytotoxicity against cultured insect cells.

Estudo de alterações estruturais cerebrais em pacientes com esquizofrenia crônica e de primeiro episódio através de imagens por ressonância magnética com morfometria baseada no voxel

Introdução: Embora alterações estruturais cerebrais na esquizofrenia venham sendo repetidamente demonstradas em estudos de ressonância magnética (RM), ainda permanece incerto se tais alterações são estáticas ou progressivas. Enquanto estudos longitudinais são tradicionalmente utilizados na avaliação da questão da progressão, estudos transversais de neuroimagem comparando diretamente pacientes com esquizofrenia crônica e de primeiro episódio a controles saudáveis têm sido bastante raros até o presente. Com o recente interesse em meganálises combinando dados multicêntricos de RM visando-se a maior poder estatístico, o presente estudo multicêntrico de morfometria baseada no voxel (VBM) foi realizado para avaliar os padrões de alterações estruturais cerebrais segundo os diferentes estágios da doença, bem como para avaliar quais (se alguma) dessas alterações se correlacionariam especificamente a moderadores clínicos potenciais, tais como exposição cumulativa a antipsicóticos, tempo de doença e gravidade da doença. Métodos: Selecionou-se uma ampla amostra de pacientes com esquizofrenia (161, sendo 99 crônicos e 62 de primeiro episódio) e controles (151) a partir de quatro estudos prévios de RM (1,5T) realizados na mesma região do Brasil. O processamento e análise das imagens foi realizado usando-se o software Statistical Parametric Mapping (SPM8) com emprego do algoritmo DARTEL (diffeomorphic anatomical registration through exponentiated Lie algebra). Os efeitos de grupo sobre os volumes regionais de substância cinzenta (SC) foram analisados através de comparações voxel-a-voxel por análises de covariância em modelos lineares gerais, inserindo-se, em todas as análises, o volume total de SC, protocolo do scanner, idade e sexo como variáveis de confusão. Por fim, foram realizadas análises de correlação entre os aludidos moderadores clínicos potenciais e os volumes cerebrais globais e regionais. Resultados: Os pacientes com esquizofrenia de primeiro episódio apresentaram reduções volumétricas sutis em comparação aos controles, em um circuito neural circunscrito e identificável apenas em análises SVC (small volume correction) [p < 0.05, com correção family-wise error (FWE)], incluindo a ínsula, estruturas têmporo-límbicas e corpo estriado. Os pacientes crônicos, por outro lado, apresentaram um padrão de alterações extensas comparativamente aos controles, envolvendo os córtices frontais orbitais, superiores e inferiores bilateralmente, córtex frontal médio direito, ambos os córtices cingulados anteriores, ambas as ínsulas, e os córtices temporais superior e médio direitos (p < 0.05, análises whole-brain com correção FWE). Foram encontradas correlações negativas significantes entre exposição cumulativa a antipsicóticos e volumes globais de SC e substância branca nos pacientes com esquizofrenia, embora as correlações com reduções regionais não tenham sido significantes. Detectaram-se, ainda, correlações negativas significantes entre tempo de doença e volumes regionais relativos da ínsula esquerda, córtex cingulado anterior direito e córtices pré-frontais dorsolaterais nas análises SVC para os grupos conjuntos (esquizofrenia crônica e de primeiro episódio). Conclusão: Os achados supracitados indicam que: a) as alterações estruturais associadas com o diagnóstico de esquizofrenia são mais disseminadas na forma crônica em comparação à de primeiro episódio; b) reduções volumétricas regionais em áreas específicas do cérebro podem variar em função do tempo de doença; c) a exposição cumulativa a antipsicóticos associou-se a alterações volumétricas globais, e não regionais

Modifying enzyme activity and selectivity by immobilization

Immobilization of enzymes may produce alterations in their observed activity, specificity or selectivity. Although in many cases an impoverishment of the enzyme properties is observed upon immobilization (caused by the distortion of the enzyme due to the interaction with the support) in some instances such properties may be enhanced by this immobilization. These alterations in enzyme properties are sometimes associated with changes in the enzyme structure. Occasionally, these variations will be positive. For example, they may be related to the stabilization of a hyperactivated form of the enzyme, like in the case of lipases immobilized on hydrophobic supports via interfacial activation. In some other instances, these improvements will be just a consequence of random modifications in the enzyme properties that in some reactions will be positive while in others may be negative. For this reason, the preparation of a library of biocatalysts as broad as possible may be a key turning point to find an immobilized biocatalyst with improved properties when compared to the free enzyme. Immobilized enzymes will be dispersed on the support surface and aggregation will no longer be possible, while the free enzyme may suffer aggregation, which greatly decreases enzyme activity. Moreover, enzyme rigidification may lead to preservation of the enzyme properties under drastic conditions in which the enzyme tends to become distorted thus decreasing its activity. Furthermore, immobilization of enzymes on a support, mainly on a porous support, may in many cases also have a positive impact on the observed enzyme behavior, not really related to structural changes. For example, the promotion of diffusional problems (e.g., pH gradients, substrate or product gradients), partition (towards or away from the enzyme environment, for substrate or products), or the blocking of some areas (e.g., reducing inhibitions) may greatly improve enzyme performance. Thus, in this tutorial review, we will try to list and explain some of the main reasons that may produce an improvement in enzyme activity, specificity or selectivity, either real or apparent, due to immobilization.

Probing the Electrocatalytic Oxygen Reduction Reaction Reactivity of Immobilized Multicopper Oxidase CueO

The bioelectrocatalytic (oxygen reduction reaction, ORR) properties of the multicopper oxidase CueO immobilized on gold electrodes were investigated. Macroscopic electrochemical techniques were combined with in situ scanning tunneling microscopy (STM) and surface-enhanced Raman spectroscopy at the ensemble and at the single-molecule level. Self-assembled monolayer of mercaptopropionic acid, cysteamine, and p-aminothiophenol were chosen as redox mediators. The highest ORR activity was observed for the protein attached to amino-terminated adlayers. In situ STM experiments revealed that the presence of oxygen causes distinct structure and electronic changes in the metallic centers of the enzyme, which determine the rate of intramolecular electron transfer and, consequently, affect the rate of electron tunneling through the protein. Complementary Raman spectroscopy experiments provided access for monitoring structural changes in the redox state of the type 1 copper center of the immobilized enzyme during the CueO-catalyzed oxygen reduction cycle. These results unequivocally demonstrate the existence of a direct electronic communication between the electrode substrate and the type 1 copper center.

Disintegrability under composting conditions of plasticized PLA–PHB blends

The disintegration under composting conditions of films based on poly(lactic acid)–poly(hydroxybutyrate) (PLA–PHB) blends and intended for food packaging was studied. Two different plasticizers, poly(ethylene glycol) (PEG) and acetyl-tri-n-butyl citrate (ATBC), were used to limit the inherent brittleness of both biopolymers. Neat PLA, plasticized PLA and PLA–PHB films were processed by melt-blending and compression molding and they were further treated under composting conditions in a laboratory-scale test at 58 ± 2 °C. Disintegration levels were evaluated by monitoring their weight loss at different times: 0, 7, 14, 21 and 28 days. Morphological changes in all formulations were followed by optical and scanning electron microscopy (SEM). The influence of plasticizers on the disintegration of PLA and PLA–PHB blends was studied by evaluating their thermal and nanomechanical properties by thermogravimetric analysis (TGA) and the nanoindentation technique, respectively. Meanwhile, structural changes were followed by Fourier transformed infrared spectroscopy (FTIR). The ability of PHB to act as nucleating agent in PLA–PHB blends slowed down the PLA disintegration, while plasticizers speeded it up. The relationship between the mesolactide to lactide forms of PLA was calculated with a Pyrolysis–Gas Chromatography–Mass Spectrometry device (Py–GC/MS), revealing that the mesolactide form increased during composting.

Oportunidad de integración de la perspectiva de género en investigación e innovación en salud en Europa: red COST genderSTE

La Comisión Europea apoya, por varias vías, incorporar la perspectiva de género, ahora en el nuevo programa Horizonte 2020, pero también financiando proyectos como gendered innovations, que muestran cómo las innovaciones de género aportan calidad en investigación y en prácticas profesionales para la salud y el bienestar. Uno de sus instrumentos políticos es la Recomendación sobre Género, Ciencia e Innovación, a desarrollar en los Estados miembros. En este contexto se crea la red internacional de Género, Ciencia, Tecnología y Medio Ambiente (COST genderSTE), que pretende: 1) cambios estructurales de las instituciones, que promocionen a las investigadoras; 2) identificación de las dimensiones de género relevantes para el medio ambiente; y 3) promocionar una mejor integración de la perspectiva de género en investigación y tecnología. COST GenderSTE apoya la creación de redes y la difusión del conocimiento con perspectiva de género. Todas estas herramientas ofrecen la oportunidad de incorporar la perspectiva de género en investigación en Europa.

La importancia del higienismo y la potabilización del agua en la ciudad de Valencia (1860-1910)

El siglo XIX se caracteriza por numerosos cambios tanto a nivel social como estructural de la población: se produce un acelerado desarrollo urbano sin planificación y carente de servicios que da lugar a una mala calidad de vida y numerosas epidemias. Estos cambios tienen como resultado un nuevo movimiento, el Higienismo, que pretende mejorar las condiciones de vida de la población. En España, todas las reformas higiénicas se desarrollaron mediante los planes de Ensanche. Uno de los puntos clave de este pensamiento es el abastecimiento de agua potable como medio para evitar enfermedades. La ciudad de Valencia es un claro ejemplo de la importancia de este movimiento y cómo el abastecimiento de agua es un elemento clave en la mejora de la calidad de vida de las personas.

Queratocone : impacto psico-social da doença na vida do doente

Trabalho Final do Curso de Mestrado Integrado em Medicina, Faculdade de Medicina, Universidade de Lisboa, 2014

The Opportunity Costs of STEM Degrees and the Unmet Needs of the Low-Skilled: Two labour market problems explained. CEPS Policy Brief No. 295, 26 June 2013

There is general consensus that to achieve employment growth, especially for vulnerable groups, it is not sufficient to simply kick-start economic growth: skills among both the high- and low-skilled population need to be improved. In particular, we argue that if the lack of graduates in science, technology, engineering and mathematics (STEM) is a true problem, it needs to be tackled via incentives and not simply via public campaigns: students are not enrolling in ‘hard-science’ subjects because the opportunity cost is very high. As far as the low-skilled population is concerned, we encourage EU and national policy-makers to invest in a more comprehensive view of this phenomenon. The ‘low-skilled’ label can hide a number of different scenarios: labour market detachment, migration, and obsolete skills that are the result of macroeconomic structural changes. For this reason lifelong learning is necessary to keep up with new technology and to shield workers from the risk of skills obsolescence and detachment from the labour market.

Many worlds of the ‘low-skilled’, but only one generic policy. CEPS Policy Brief No. 312, 10 January 2014

This paper, which draws on research findings from the NEUJOBS project, encourages EU and national policy-makers to invest in a more comprehensive view of the phenomenon of ‘low-skilledness’. The ‘low-skilled’ label can hide a number of different scenarios: labour market detachment, migration and obsolete skills that are the result of macroeconomic structural changes. For this reason, the authors argue that it is necessary to promote lifelong learning to allow workers to keep pace with new technologies and to shield workers from the risk of skills obsolescence and detachment from the labour market.

Neospora caninum calcium-dependent protein kinase 1 is an effective drug target for neosporosis therapy.

Despite the enormous economic importance of Neospora caninum related veterinary diseases, the number of effective therapeutic agents is relatively small. Development of new therapeutic strategies to combat the economic impact of neosporosis remains an important scientific endeavor. This study demonstrates molecular, structural and phenotypic evidence that N. caninum calcium-dependent protein kinase 1 (NcCDPK1) is a promising molecular target for neosporosis drug development. Recombinant NcCDPK1 was expressed, purified and screened against a select group of bumped kinase inhibitors (BKIs) previously shown to have low IC50s against Toxoplasma gondii CDPK1 and T. gondii tachyzoites. NcCDPK1 was inhibited by low concentrations of BKIs. The three-dimensional structure of NcCDPK1 in complex with BKIs was studied crystallographically. The BKI-NcCDPK1 structures demonstrated the structural basis for potency and selectivity. Calcium-dependent conformational changes in solution as characterized by small-angle X-ray scattering are consistent with previous structures in low Calcium-state but different in the Calcium-bound active state than predicted by X-ray crystallography. BKIs effectively inhibited N. caninum tachyzoite proliferation in vitro. Electron microscopic analysis of N. caninum cells revealed ultra-structural changes in the presence of BKI compound 1294. BKI compound 1294 interfered with an early step in Neospora tachyzoite host cell invasion and egress. Prolonged incubation in the presence of 1294 interfered produced observable interference with viability and replication. Oral dosing of BKI compound 1294 at 50 mg/kg for 5 days in established murine neosporosis resulted in a 10-fold reduced cerebral parasite burden compared to untreated control. Further experiments are needed to determine the PK, optimal dosage, and duration for effective treatment in cattle and dogs, but these data demonstrate proof-of-concept for BKIs, and 1294 specifically, for therapy of bovine and canine neosporosis.

A new institutional approach to Japanese firms' foreign direct investment under free trade agreements

This paper examines the determinants of foreign direct investment (FDI) under free trade agreements (FTAs) from a new institutional perspective. First, the determinants of FDI are theoretically discussed from a new institutional perspective. Then, FDI is statistically analyzed at the aggregate level. Kernel density estimation of firm-size reveals some evidence of "structural changes" after FTAs, as characterized by the investing firms' paid-up capital stock. Statistical tests of the average and variance of the size distribution confirm this in the case of FTAs with Asian partner countries. For FTAs with South American partner countries, the presence of FTAs seems to promote larger-scale FDIs. These results remain correlational instead of causal, and more statistical analyses would be needed to infer causality. Policy implications suggest that participants should consider "institutional" aspects of FTAs, that is, the size matters as a determinant of FDI. Future work along this line is needed to study "firm heterogeneity."

Appendix A Table SD1. CaCO3, d18O, d13C and Sr isotopes from ODP Site 181-1120

An integrated chemostratigraphic (87Sr/86Sr, d13C and 18O) study of benthic foraminifera is presented for a 210 m-thick, intermediate depth (upper/middle bathyal transition), Miocene nannofossil ooze section of Ocean Drilling Program Site 1120, Campbell Plateau off New Zealand. Our results indicate that new 87Sr/86Sr, d13C and d18O profiles are wholly consistent with their respective Miocene reference curves. These observations facilitate identification of a total of five reliable chemostratigraphic datums, which are based on the fundamental structural changes in the 87Sr/86Sr curve and paired simultaneous d13C and d18O events. The resultant age-depth relationship clearly shows that the Miocene (20-5 Ma) biopelagic sedimentation on the Campbell Plateau was essentially continuous at a moderate to high, linear sedimentation rate (17.5 m/m.y. with an exception of the uppermost 13 m). Our findings do not support the shipboard biostratigraphic age model, which assumes that the critical early-middle Miocene transition was interrupted by a major hiatus (<~3 m.y.). Because of its unique bathymetric setting at a paleowater-depth of ~ 600 m, which is among the shallowest of the coeval isotopically studied deep-sea sections in the South Pacific/Southern Ocean, Site 1120 will serve as a reference section for surveying the evolution of intermediate-water paleoceanography in the Southern Hemisphere across the middle Miocene climatic transition.

The evolution of microRNA in primates

Thesis (Ph.D.)--University of Washington, 2016-06