Sentido de coerência : ligação materno-fetal e estilo de vida da mulher durante a gravidez

RESUMO - Um dos grandes desafios actuais enfrentados pela Saúde Pública diz respeito ao fardo representado pelas doenças crónicas não transmissíveis enquanto co-responsáveis pela maioria das mortes que ocorrem no mundo, pela significativa e progressiva redução da qualidade de vida e aumento das incapacidades dos indivíduos afectados e por uma fasquia bastante elevada das despesas em saúde. Entretanto, a complexa dinâmica genética, biológica, psicológica, afectiva, sócio-cultural e ambiental que envolve o comportamento humano, tão amplamente relacionado com algumas destas doenças – doenças cardiovasculares, alguns tipos de cancro, obesidade, hipertensão, diabetes e doenças osteo-articulares – impõe o desafio constante da busca de novas e efectivas intervenções em promoção da saúde que influenciem positivamente os estilos de vida dos indivíduos, dos grupos e das comunidades. Sendo o sentido de coerência um traço da personalidade do indivíduo desenvolvido sob a influência directa ou indirecta dos mesmo factores acima referidos, o estabelecimento de uma relação entre este constructo e os comportamentos humanos pode revelar-se promissor para a elaboração de novas intervenções em promoção da saúde. Por sua vez, a gravidez, talvez por influência da ligação materno-fetal, pode representar um ponto de viragem na vida da mulher no que respeita ao sentido de coerência e aos comportamentos de saúde e um bom começo na vida do bebé que irá nascer sob a influência dos mesmos. Com a finalidade de contribuir para a construção de intervenções efectivas em promoção da saúde, através da descoberta de prováveis potencialidades salutogénicas dos constructos referidos – sentido de coerência e ligação materno-fetal –, foi desenvolvido um estudo quantitativo, observacional que teve por objectivos: ampliar o conhecimento sobre as mudanças do sentido de coerência no decorrer da vida, especificamente durante a gravidez; ampliar o conhecimento acerca das relações de alguns factores de natureza sócio-demográfica, psico-afectiva e obstétrica com o sentido de coerência das mulheres grávidas e com a ligação materno-fetal; e identificar possíveis relações entre o sentido de coerência, a ligação materno-fetal e o estilo de vida das mulheres grávidas, este último representado pelos hábitos alimentares, consumo de cafeína, consumo de álcool, hábitos tabágicos, prática regular de actividade física e ganho de peso durante a gravidez. O presente relatório descreve a concepção e os resultados deste estudo, que envolveu a uma amostra de 61 mulheres grávidas que estavam a ser acompanhadas nos serviços de saúde materna dos Centros de Saúde de Carnaxide extensão Linda-a-Velha e de Oeiras, no Distrito de Lisboa, Portugal, entre os meses de Fevereiro e Julho de 2005. Os resultados demonstram que, para a amostra de 61 mulheres grávidas que responderam ao inquérito por questionário de auto-resposta, foram encontradas associações estatisticamente significativas entre o sentido de coerência e a escolaridade e entre o sentido de coerência e a percepção do próprio estado de saúde. Além disso, foi encontrada alguma evidência das associações entre o sentido de coerência e a faixa etária, estado civil e rendimento mensal familiar e foi observada alguma tendência para que as mulheres grávidas com níveis de sentido de coerência mais elevados consumissem menos álcool do que as mulheres grávidas com níveis de sentido de coerência inferiores. Entretanto, as demais associações testadas não foram confirmadas. Relativamente à ligação materno-fetal, foram encontradas, para a amostra de 41 mulheres grávidas que participaram do segundo momento de colheita de dados do estudo, entre a 20ª e a 24ª semanas de gravidez, associações estatisticamente significativas com a escolaridade e o nível de rendimento familiar das mulheres grávidas, não tendo sido confirmadas as demais associações testadas. Embora não tenham sido estatisticamente evidenciadas as relações entre o sentido de coerência e a ligação materno-fetal e entre estes e os comportamentos de saúde, o carácter preliminar destes resultados, devido à subjectividade do processo de selecção não probabilístico da amostra estudada e à reduzida dimensão desta amostra, e a escassez de estudos descritos na literatura fazem com que seja prudente a realização de estudos de follow-up, com amostras de maiores dimensões, aleatórias e representativas da população, para que sejam estabelecidas quaisquer conclusões acerca destas questões.-----------------------------ABSTRACT - One of the greatest challenges faced by Public Health in nowadays is the burden represented by chronic diseases as co-responsible for the majority of deaths that occurs in the world, for the meaningful and progressive reduction of quality of life and increase of disabilities in affected individuals and for an important part of health expenses. However, the complexity of the genetic, biological, psychological, emotional, social, cultural and environmental dynamics that involves human behaviours related to some of these diseases – cardiovascular diseases, some kind of cancers, obesity, hypertension, diabetes and joint and bone diseases – poses the continuous challenge of searching for new and effective interventions of health promotion that positively influence individuals, groups and community lifestyles. Due to the fact that sense of coherence is an individual personality trace directly or indirectly influenced by the same factors listed above, the discovery of a relationship between this construct and human behaviours might be promising to the creation of new health promotion interventions. On the other hand, pregnancy may represent a turn point to the mother’s life and a good start in the baby’s life in relation to sense of coherence and health behaviours and It might occur because of the influence of maternal-fetal attachment. With the purpose of contributing with the creation of effective health promotion interventions through the discovery of probable salutogenic potentials in the referred constructs – sense of coherence and maternal-fetal attachment – , it was developed a quantitative observational study with the following objectives: to increase knowledge about changes in sense of coherence throughout life, specifically during pregnancy; to increase knowledge about the relationship between sense of coherence and maternal-fetal attachment and some social, demographical, psychological, emotional and obstetric factors of pregnant women; to identify probable relationships between sense of coherence, maternal-fetal attachment and pregnant women’s lifestyles, represented by diet habits, caffeine consumption, alcohol consumption, smoking habits, physical activity habits and weigh gain during pregnancy. This report describes the structure and the findings of this study involving a sample of 61 pregnant women who had been followed by health professionals in the mother health services of Carnaxide (Linda-a-Velha unity) and Oeiras Health Centres, in Lisbon, Portugal, between February and July of 2005. The results show that, for the 61 pregnant women who filled the self-reported questionnaire, it was found a statistically significant association between sense of coherence and education level. It was also found some evidence of the associations between sense of coherence and age, marital status and mensal household income and a trend toward pregnant women with higher levels of sense of coherence to drink less alcoholic beverages than pregnant women with lower levels of sense of coherence. However, the others associations tested were not confirmed. Regarding maternal-fetal attachment, it was found, for the sample of 41 women who participated in the second moment of data collection, between the 20th and the 24th week of pregnancy, statistically significant associations with education level and mensal household income. The others associations tested were not confirmed. Although the associations between sense of coherence and maternal-fetal attachment and between these constructs and health behaviours were not confirmed, all findings presented here are considered preliminary because of small dimension of sample and non-probabilistic criteria used for sample selection. What’s more, there are almost no studies described in the literature which could confirm or contradict these findings. Therefore, it is better to be careful and develop follow-up studies, with bigger and representative of population samples, before draw any conclusions about these theme.

Evolution of Drug Resistance: Insight on TEM β-Lactamases Structure and Activity and β-Lactam Antibiotics

Since the discovery of the first penicillin bacterial resistance to β-lactam antibiotics has spread and evolved promoting new resistances to pathogens. The most common mechanism of resistance is the production of β-lactamases that have spread thorough nature and evolve to complex phenotypes like CMT type enzymes. New antibiotics have been introduced in clinical practice, and therefore it becomes necessary a concise summary about their molecular targets, specific use and other properties. β-lactamases are still a major medical concern and they have been extensively studied and described in the scientific literature. Several authors agree that Glu166 should be the general base and Ser70 should perform the nucleophilic attack to the carbon of the carbonyl group of the β-lactam ring. Nevertheless there still is controversy on their catalytic mechanism. TEMs evolve at incredible pace presenting more complex phenotypes due to their tolerance to mutations. These mutations lead to an increasing need of novel, stronger and more specific and stable antibiotics. The present review summarizes key structural, molecular and functional aspects of ESBL, IRT and CMT TEM β-lactamases properties and up to date diagrams of the TEM variants with defined phenotype. The activity and structural characteristics of several available TEMs in the NCBI-PDB are presented, as well as the relation of the various mutated residues and their specific properties and some previously proposed catalytic mechanisms.

The social stock exchange: A quantitative exploration

A Work Project, presented as part of the requirements for the Award of a Masters Degree in Economics from the NOVA – School of Business and Economics

Relationship between galactomannan structure and physicochemical properties of films produced thereof

In this work five sources of galactomannans, Adenanthera pavonina, Cyamopsis tetragonolobus, Caesalpinia pulcherrima, Ceratonia siliqua and Sophora japonica, presenting mannose/galactose ratios of 1.3, 1.7, 2.9, 3.4 and 5.6, respectively, were used to produce galactomannan-based films. These films were characterized in terms of: water vapour, oxygen and carbon dioxide permeabilities (WVP, O 2 P and CO 2 P); moisture content, water solubility, contact angle, elongation-at-break (EB), tensile strength (TS) and glass transition temperature (T g ). Results showed that films properties vary according to the galactomannan source (different galactose distribution) and their mannose/galactose ratio. Water affinity of mannan and galactose chains and the intermolecular interactions of mannose backbone should also be considered being factors that affect films properties. This work has shown that knowing mannose/galactose ratio of galactomannans is possible to foresee galactomannan-based edible films properties.

Body image dissatisfaction and its relationship with physical activity and body mass index in Brazilian adolescents

Objective: To evaluate body image dissatisfaction and its relationship with physical activity and body mass index in a Brazilian sample of adolescents. Methods: A total of 275 adolescents (139 boys and 136 girls) between the ages of 14 and 18 years completed measures of body image dissatisfaction through the Contour Drawing Scale and current physical activity by the International Physical Activity Questionnaire. Weight and height were also measured for subsequent calculation of body mass index. Results: Boys and girls differed significantly regarding body image dissatisfaction, with girls reporting higher levels of dissatisfaction. Underweight and eutrophic boys preferred to be heavier, while those overweight preferred be thinner and, in contrast, girls desired to be thinner even when they are of normal weight. Conclusion: Body image dissatisfaction was strictly related to body mass index, but not to physical activity.

Poly(ester-urethane) scaffolds: effect of structure on properties and osteogenic activity of stem cells

The present study aimed to investigate the effect of structure (design and porosity) on the matrix stiffness and osteogenic activity of stem cells cultured on poly(ester-urethane) (PEU) scaffolds. Different three-dimensional (3D) forms of scaffold were prepared from lysine-based PEU using traditional salt-leaching and advanced bioplotting techniques. The resulting scaffolds were characterized by differential scanning calorimetry (DSC), thermogravimetric analysis (TGA), scanning electron microscopy (SEM), mercury porosimetry and mechanical testing. The scaffolds had various pore sizes with different designs, and all were thermally stable up to 300â °C. In vitrotests, carried out using rat bone marrow stem cells (BMSCs) for bone tissue engineering, demonstrated better viability and higher cell proliferation on bioplotted scaffolds compared to salt-leached ones, most probably due to their larger and interconnected pores and stiffer nature, as shown by higher compressive moduli, which were measured by compression testing. Similarly, SEM, von Kossa staining and EDX analyses indicated higher amounts of calcium deposition on bioplotted scaffolds during cell culture. It was concluded that the design with larger interconnected porosity and stiffness has an effect on the osteogenic activity of the stem cells.

Regulación transcripcional y relación entre estructura y función de proteínas relacionadas con el metabolismo de colina en Pseudomonas aeruginosa. Transcriptional regulation and relationship between structure and function of proteins related with the choline metabolism in Pseudomonas aeruginosa.

El objetivo general de este proyecto es dilucidar los mecanismos de acción a nivel molecular de enzimas y proteínas involucradas en el metabolismo de colina en Pseudomonas aeruginosa, con énfasis en la identificación de residuos aminoacídicos críticos y regulación de la expresión de los genes en estudio. Los objetivos específicos que se palntean involucran abordajes bioquímicos y moleculares y serán llevados a cabo mediante técnicas de biología molecular y bioquímica (mutación sitio-dirigida, deleción génica, expresión y purificación de proteínas, fusión transcripcional a genes reporteros, etc). Planteo de hipótesis: las proteínas que se inducen por colina (fosforilcolina fosfatasa (PchP), fosfolipasa C (PlcH), acetilcolinestera (AchE), proteínas periplásmicas unidoras de colina (PUch) podrían compartir: a) una organización génica y responder a la regulación por proteínas regulatorias o a factores ambientales de manera similar; b) residuos aminoacídicos conservados que intervengan en la unión o interacción con diferentes ligandos, principalmente, colina. Para ello, se plantean los siguientes Objetivos Específicos: 1) identificar las zonas promotoras de los genes que codifican para PchP, PlcH, AchE y PUch, a fin de localizar posibles sitios de unión a proteínas reguladoras y los factores ambientales que afectan la actividad promotora. 2) determinar en las proteínas mencionadas los residuos aminoacídicos de importancia involucrados en la catálisis y en la interacción con ligandos, principalmente en la unión a compuestos de alquilamonio; 3) Se iniciarán estudios que demuestren la relación entre la inducción por colina de varios factores de patogenicidad la virulencia del microorganismo, empleando mutantes simples o múltiples en estos factores y como modelo de patogenicidad el nematodo C. elegans. A partir de los resultados obtenidos se pretende tener un conocimiento profundo sobre la regulación molecular y bioquímica de varias enzimas comprometidas en la patología que produce P. aeruginosa. Esto más el conocimiento de la fisiología de este microorganismo abre el camino para la búsqueda de posibles blancos de acción de drogas. Por otro lado, se espera tener un conocimiento integral sobre la regulación de la expresión de las actividades enzimáticas relacionadas con el metabolismo de colina y la respuesta de P. aeruginosa ante la presencia de compuestos de alquilamonio utilizados como nutrientes. Se espera conocer el papel que desempeña cada uno de los sitios de unión a los diferentes ligandos para el funcionamiento y control de las enzimas mencionadas y explicar el comportamiento diferencial de las enzimas frente a distintos sustratos y otros ligandos. El conocimiento de los sitios de unión a compuestos de alquilamonio permitirá encontrar esos dominios en diferentes proteínas del género Pseudomonas y otras bacterias Gram negativas. Desde el punto de vista evolutivo, se podrá comparar la similitud de los sitios de unión a colina entre proteínas de organismos eucariotas con procariotas (ej. PUch de bacterias Gram positivas, transportadores de colina, proteína C reactiva, AchE de eucariotas contra las encontradas en bacterias del género Pseudomonas, fosfolipasas A, C o D, etc.). Este proyecto permitirá concretar al menos dos tesis doctorales (Sanchez, Otero) más varios trabajos finales de grado (tesinas) que son y serán realizados por alumnos de la carrera de Microbiología en la UNRC. Les permitirá a los doctorandos y a los alumnos de grado adquirir una formación bastante integral ya que utilizarán herramientas de la fisiología general bacteriana, de la bioquímica clásica, de la biología molecular y de la bioinformática.

Evaluación del efecto del láser y magnetoterapia en miopatía experimental valorando biomarcadores de estrés oxidativo, histomorfometría y actividad enzimática mitocondrial. Evaluation of the effect of laser and magnetic therapy in experimental myopathy assessing oxidative stress biomarkers, histomorphometry and mitochondrial enzyme activity

El láser de baja y media energía y la magnetoterapia son utilizados en desórdenes osteomioarticulares por sus efectos analgésico, antiinflamatorio y trófico, entre los más destacados. Sin embargo, son insuficientes las investigaciones sobre su mecanismo de acción y antecedentes científicos que avalen sus efectos. Es por ello, que la determinación de acontecimientos celulares y moleculares que ocurren durante la interacción de estos tipos de energía con el sistema muscular, sería relevante para el conocimiento y optimización de tales terapias en las ciencias biomédicas. En las miopatías inflamatorias idiopáticas, se encuentra afectada la estructura, morfología y bioquímica del tejido muscular. La energía que éste requiere para el normal funcionamiento es generada en la mitocondria. Esta organela también es la responsable de la generación de especies oxidantes provocando estrés oxidativo y el inicio de los procesos de apoptosis. Por lo antes dicho, consideramos que la determinación de los biomarcadores inflamatorios asociados a estrés oxidativo, realizando el análisis histomorfométrico ultraestructural y valorando la actividad de los complejos enzimáticos mitocondriales, permitiría una evaluación de la acción terapéutica del láser y la magnetoterapia en un modelo experimental de miopatía. Para ello se propone evaluar el efecto de la magnetoterapia y del láser de baja energía (He-Ne y As.Ga) en miopatía experimental determinando indicadores inflamatorios asociados a estrés oxidativo, análisis histomorfométrico y valoración de la actividad enzimática mitocondrial. Específicamente: -Determinar indicadores inflamatorios y de estrés oxidativo: Oxido Nítrico, Grupos carbonilos, L-citrulina, Fibrinógeno, Superóxido dismutasa, Glutation peroxidasa y Catalasa por espectrofotometría. -Identificar los cambios anatomopatológicos del músculo esquelético por microscopía óptica (MO): cuantificación del infiltrado inflamatorio; MO de alta resolución (MOAR) y por microscopía electrónica: histomorfometría de la ultraestructura miofibrilar y mitocondrial. -Valorar las actividades enzimáticas de la citrato sintasa y de los complejos: I (NADH-ubiquinona reductasa), II (succinato-ubiquinona-reductasa) III (ubiquinona-citocromo c-reductasa) y IV (citocromo c-oxidasa); en mitocondrias de tejido muscular por espectrofotometría. -Evaluar la actividad apoptótica en las fibras musculares de los diferentes grupos por ténica de T.U.N.E.L. Las mediciones mitocondriales (por ME) y de infiltrado inflamatorio (por MO) se realizarán en un total de 5 fotos de aumentos similares en forma aleatoria por grupo estudiado (n=10). Los cambios estructurales observados se analizarán en el programa Axiovision 4.8, para cuantificar el área total ocupada, número total y grado de alteración de las mitocondrias y el porcentaje de infiltrado inflamatorio determinando el grado de inflamación. Los resultados de los datos cuantitativos se analizarán aplicando ANAVA (test de Fisher para comparaciones múltiples); y para los datos categóricos se utilizará Chi cuadrado (test de Pearson), estableciéndose un nivel de significación de p < 0.05 para todos los casos. Importancia del Proyecto: La salud y el bienestar del hombre son los logros perseguidos por las ciencias de la salud. La obtención de terapias curativas o paliativas con un mínimo de efectos colaterales para el enfermo se incluye en estos logros. Por esto y todo lo anteriormente expuesto es que consideramos de gran importancia poder esclarecer desde las ciencias básicas los efectos celulares y moleculares en modelos experimentales la acción de la terapia con láser y magnetoterapia para una aplicación clínica con base científica en todas las áreas de las Ciencias Médicas. In the idiopathic inflammatory myopathies, is affected the structure, morphology and biochemistry of muscle tissue. The mitochondria is responsible for the generation of oxidizing species leading to oxidative stress and the beginning of the process of apoptosis. As said before, we consider the determination of inflammatory biomarkers related to oxidative stress, by ultrastructural morphometric analysis and assessing the activity of mitochondrial enzyme complexes, permit an evaluation of the therapeutic action of laser and magnetic therapy in an experimental model myopathy. We propose to evaluate the effect of the treatment identifying indicators in experimental inflammatory myopathy associated with oxidative stress, histomorphometric analysis and assessment of mitochondrial enzyme activity. Specifically -determining: Nitric oxide, carbonyl groups, L-citrulline, fibrinogen, superoxide dismutase, glutathione peroxidase and catalase by spectrophotometry. -Identify the pathological changes in skeletal muscle by optical microscopy (OM): quantification of the inflammatory infiltrate, OM high resolution (MOAR) and electron microscopy, histomorphometry of myofibrillar and mitochondrial ultrastructure. -Evaluate the enzymatic activity of citrate synthase and complexes: I, II, III and IV in mitochondria muscle tissue by spectrophotometry. -Evaluate apoptotic activity in muscle fibers by TUNEL technique of Mitochondrial measurements and inflammatory infiltration (by OM) was performed in a total of 5 photos of similar increases in random by the study group (n = 10). The structural changes observed are discussed in the program Axiovision 4.8, to quantify number, degree of alteration of mitochondria and the percentage of inflammatory infiltrate determining the degree of inflammation. The results of the quantitative data were analyzed using ANOVA (Fisher test), and categorical data with Chi-square (Pearson test), establishing a significance level of p <0.05.

Synthesis, structure and catalytic activity of new lanthanide alkynylamidinates

Magdeburg, Univ., Fak. für Verfahrens- und Systemtechnik, Diss., 2015

Thermal biology, activity, and population parameters of Cnemidophorus vacariensis (Squamata, Teiidae), a lizard endemic to southern Brazil

We investigated the following aspects of the biology of a population of Cnemidophorus vacariensis Feltrim & Lema, 2000 during the four seasons: thermal biology, relationship with the thermal environment, daily and seasonal activity, population structure and growth rate. Cnemidophorus vacariensis is restricted to rocky outcrops of the "campos de cima da serra" grasslands on the Araucaria Plateau, southern Brazil, and is currently listed as regionally and nationally threatened with extinction. Data were collected from October 2004 through September 2007 in the state of Rio Grande do Sul. Sampling was conducted randomly from 08:00 a.m. to 6:00 p.m. The capture-mark-recapture method was employed. The lizards were captured by hand, and their cloacal temperature, sex, snout-ventral length (SVL), mass, and the temperature of their microhabitat (substrate temperature and air temperature) were recorded. Individuals were then marked by toe-clipping and released at the site of capture. Body temperatures were obtained for 175 individuals, activity data for 96 individuals, and data on population structure and growth for 59 individuals. All data were obtained monthly, at different times of the day. Cnemidophorus vacariensis average body temperature was 23.84ºC, ranging between 9.6 and 38.2ºC. Temperatures ranged between 21 and 29ºC. The correlation between external heat sources, substrate and air were positive and significant and there was a greater correlation between lizard's temperature and the temperature of the substrate (tigmothermic species). The relatively low body temperatures of individuals are associated with the climate of their environment (altitude up to 1,400 m), with large variations in temperature throughout the day and the year, and low temperatures in winter. The average body temperature observed for C. vacariensis was low when compared with that of phylogenetically related species, suggesting that the thermal biology of this species reflects adaptations to the temperate region where it lives. The monthly rates of activity of lizards were related to monthly variations in the ambient temperatures. Our data suggest that the daily and seasonal activity of C. vacariensis result from the interaction between two factors: changes in the environment temperature and the relationship between individuals and their thermal environment. The population structure of C. vacariensis varied throughout the study period, with maximum biomass in January and maximum density in February (recruitment period). The sex ratio diverged from the expected 1:1. The growth analysis showed a negative relationship between the growth rate of individuals and the SVL, revealing that young individuals grow faster than adults, a typical pattern for short-lived species. The population studied showed a seasonal and cyclical variation associated with the reproductive cycle. The life strategy of C. vacariensis seems to include adaptations to the seasonal variations in temperature, typical of its environment.

Relationship of physical activity with motor skills, aerobic fitness and body fat in preschool children: a cross-sectional and longitudinal study (Ballabeina).

Adiposity, low aerobic fitness and low levels of activity are all associated with clustered cardiovascular disease risk in children and their high prevalence represents a major public health concern. The aim of this study is to investigate the relationship of objectively measured physical activity (PA) with motor skills (agility and balance), aerobic fitness and %body fat in young children. This study is a cross-sectional and longitudinal analyses using mixed linear models. Longitudinal data were adjusted for baseline outcome parameters. In all, 217 healthy preschool children (age 4-6 years, 48% boys) participated in this study. PA (accelerometers), agility (obstacle course), dynamic balance (balance beam), aerobic fitness (20-m shuttle run) and %body fat (bioelectric impedance) at baseline and 9 months later. PA was positively associated with both motor skills and aerobic fitness at baseline as well as with their longitudinal changes. Specifically, only vigorous, but not total or moderate PA, was related to changes in aerobic fitness. Higher PA was associated with less %body fat at baseline, but not with its change. Conversely, baseline motor skills, aerobic fitness or %body fat were not related to changes in PA. In young children, baseline PA was associated with improvements in motor skills and in aerobic fitness, an important determinant of cardiovascular risk.

Cysteine 230 is essential for the structure and activity of the cytotoxic ligand TRAIL.

Unlike other tumor necrosis factor family members, the cytotoxic ligand tumor necrosis factor-related apoptosis-inducing ligand (TRAIL)/Apo-2L contains an unpaired cysteine residue (Cys(230)) in its receptor-binding domain. Here we show that the biological activity of both soluble recombinant TRAIL and cell-associated, full-length TRAIL is critically dependent on the presence of Cys(230). Mutation of Cys(230) to alanine or serine strongly affected its ability to kill target cells. Binding to its receptors was decreased by at least 200-fold, and the stability of its trimeric structure was reduced. In recombinant TRAIL, Cys(230) was found engaged either in interchain disulfide bridge formation, resulting in poorly active TRAIL, or in the chelation of one zinc atom per TRAIL trimer in the active, pro-apoptotic form of TRAIL.

Cell surface expression of the epithelial Na channel and a mutant causing Liddle syndrome: a quantitative approach.

The epithelial amiloride-sensitive sodium channel (ENaC) controls transepithelial Na+ movement in Na(+)-transporting epithelia and is associated with Liddle syndrome, an autosomal dominant form of salt-sensitive hypertension. Detailed analysis of ENaC channel properties and the functional consequences of mutations causing Liddle syndrome has been, so far, limited by lack of a method allowing specific and quantitative detection of cell-surface-expressed ENaC. We have developed a quantitative assay based on the binding of 125I-labeled M2 anti-FLAG monoclonal antibody (M2Ab*) directed against a FLAG reporter epitope introduced in the extracellular loop of each of the alpha, beta, and gamma ENaC subunits. Insertion of the FLAG epitope into ENaC sequences did not change its functional and pharmacological properties. The binding specificity and affinity (Kd = 3 nM) allowed us to correlate in individual Xenopus oocytes the macroscopic amiloride-sensitive sodium current (INa) with the number of ENaC wild-type and mutant subunits expressed at the cell surface. These experiments demonstrate that: (i) only heteromultimeric channels made of alpha, beta, and gamma ENaC subunits are maximally and efficiently expressed at the cell surface; (ii) the overall ENaC open probability is one order of magnitude lower than previously observed in single-channel recordings; (iii) the mutation causing Liddle syndrome (beta R564stop) enhances channel activity by two mechanisms, i.e., by increasing ENaC cell surface expression and by changing channel open probability. This quantitative approach provides new insights on the molecular mechanisms underlying one form of salt-sensitive hypertension.

Intergenic and external transcribed spacers of ribosomal RNA genes in lizard-infecting Leishmania: molecular structure and phylogenetic relationship to mammal-infecting Leishmania in the subgenus Leishmania (Leishmania)

To establish the relationships of the lizard- and mammal-infecting Leishmania, we characterized the intergenic spacer region of ribosomal RNA genes from L. tarentolae and L. hoogstraali. The organization of these regions is similar to those of other eukaryotes. The intergenic spacer region was approximately 4 kb in L. tarentolae and 5.5 kb in L. hoogstraali. The size difference was due to a greater number of 63-bp repetitive elements in the latter species. This region also contained another element, repeated twice, that had an inverted octanucleotide with the potential to form a stem-loop structure that could be involved in transcription termination or processing events. The ribosomal RNA gene localization showed a distinct pattern with one chromosomal band (2.2 Mb) for L. tarentolae and two (1.5 and 1.3 Mb) for L. hoogstraali. The study also showed sequence differences in the external transcribed region that could be used to distinguish lizard Leishmania from the mammalian Leishmania. The intergenic spacer region structure features found among Leishmania species indicated that lizard and mammalian Leishmania are closely related and support the inclusion of lizard-infecting species into the subgenus Sauroleishmania proposed by Saf'janova in 1982.

Nutritional status using the Mini Nutritional Assessment questionnaire and its relationship with bone quality in a population of institutionalized elderly women.

Malnutrition, a risk factor for osteoporotic fractures, is frequent in elderly people and, is underdiagnosed and undertreated. There are only few studies on the nutritional status of elderly people in Europe. The Mini Nutritional Assessment (MNA) is a non invasive and validated questionnaire to evaluate nutritional status in elderly people, classified in three groups: 1 degree score < 17: malnourished, 2 degrees score >17 and < 24: at risk of malnutrition, 3 degrees score >24: well-nourished, with a maximum of 30 points. Quantitative ultrasound of bone (QUS) is a method for assessing quality of bone which can be easily performed in nursing homes. Therefore, these two tests allowed to study the relationships between nutritional status and ultrasonic parameters of bone in 78 institutionalized women aged 86 +/- 6 years, living in 11 nursing homes around Lausanne (Switzerland). All were assessed by the MNA, had a measurement of the tricipital skin fold and of the grip strength. Functional status was evaluated by the scale "Activity of Daily Living" (ADL), and serum albumin level was measured when permitted. All had QUS of the calcaneus (with an Achilles, GE Lunar). The measured parameters are the Broadband Ultrasound Attenuation (BUA), attenuation of a band of ultrasonic frequencies through the medium, expressed in dB/MHz, and the Speed of Sound (SOS), speed of the ultrasounds through the medium, expressed in m/s. A third parameter, the stiffness index (SI), expressed as a percentage of the values obtained by the manufacturer in a young population and derived from BUA and SOS, was calculated automatically : SI = (0.67xBUA) + (0.28xSOS) - 420, expressed in percent compared to a young adult population (%YA). Fifteen percent of the women were undernourished and 58% were at risk of malnutrition. As expected, compared with the well-nourished minority, undernourished subjects had significant lower body mass index (BMI), tricipital skin fold (TSF), ADL score and albumin level (p < 0,01). The subjects "at risk of malnutrition" had significant lower BMI, ADL score (p < 0.01), tricipital skin fold and serum albumin (p < 0.05). Ultrasound parameters were low independently of the nutritional status. MNA score correlated significantly with tricipital skin fold (r = 0.508, p < 0.01), ADL (r = 0.538, p < 0.01) and albumin serum level (r = 0.409, p = 0.01). There was a trend for a correlation between the MNA and the ultrasound parameter BUA (r = 0.207, p = 0.07), whereas no correlation was found with SOS and SI. A multivariate analysis showed that tricipital skin fold and ADL explained 61% of the variance of the MNA. In conclusion, using simple and non invasive methods, this study showed that malnutrition and osteoporosis are frequent in institutionalized elderly persons in our country, and the ultrasound parameters are influenced by many others factors in addition to nutrition, especially at this age and in elderly residents of nursing homes.