Functional significance of a highly conserved glutamate residue of the human noradrenaline transportere

The aim of the study was to investigate the role of glutamate residue 113 in transmembrane domain 2 of the human noradrenaline transporter in determining cell surface expression and functional activity. This residue is absolutely conserved in all members of the Na+- and Cl--dependent transporter family. Mutations to alanine (hE113A), aspartate (hE113D) and glutamine (hE113Q) were achieved by site-directed mutagenesis and the mutants were expressed in transfected COS-7 or HEK-293 cells. Cell surface expression of IIE113A and hE113D, but not hE113Q, was markedly reduced compared with wild type, and functional noradrenaline uptake was detected only for the hE113Q mutant. The pharmacological properties of the hE113Q mutant showed very little change compared with wild type, except for a decrease in V-max values for noradrenaline and dopamine uptake of 2-3-fold. However, the hE113D mutant showed very marked changes in its properties, compared with wild type, with 82-260-fold decreases in the affinities of the substrates, noradrenaline, dopamine and MPP+, and increased Na+ affinity for stimulation of nisoxetine binding. The results of the study show that the size and not the charge of the 113 glutamate residue of the noradrenaline transporter seems to be the most critical factor for maintenance of transporter function and surface expression.

Heterogeneity in olfactory neurons in mouse revealed by differential expression of glycoconjugates

Cell surface glycoconjugates have been implicated in the growth and guidance of subpopulations of primary olfactory axons. While subpopulations of primary olfactory neurons have been identified by differential expression of carbohydrates in the rat there are few reports of similar subpopulations in the mouse. We have examined the spatiotemporal expression pattern of glycoconjugates recognized by the lectin from Wisteria floribunda (WFA) in the mouse olfactory system. In the developing olfactory neuroepithelium lining the nasal cavity, WFA stained a subpopulation of primary olfactory neurons and the fascicles of axons projecting to the target tissue, the olfactory bulb. Within the developing olfactory bulb, WFA stained the synaptic neuropil of the glomerular and external plexiform layers. In adults, strong expression of WFA ligands was observed in second-order olfactory neurons as well as in neurons in several higher order olfactory processing centres in the brain. Similar, although distinct, staining of neurons in the olfactory pathway was detected with Dolichos biflorus agglutinin. These results demonstrate that unique subpopulations of olfactory neurons are chemically coded by the expression of glycoconjugates. The conserved expression of these carbohydrates across species suggests they play an important role in the functional organization of this region of the nervous system.

ADH genotype does not modify the effects of alcohol on high-density lipoprotein

Background: Alcohol consumption has beneficial effects on mortality which are mainly due to reduction in cardiovascular disease. These are believed to be due, at least in part, to the increase in plasma high-density lipoprotein (HDL) which is associated with alcohol consumption. It has been proposed that ADH3 genotype modifies the relationships between alcohol intake and cardiovascular disease by altering the HDL response to alcohol. The aim of this paper was to test for effects of ADH2 and ADH3 genotypes on the response of HDL components to habitual alcohol consumption. Methods: Adult male and female subjects were genotyped for ADH2 and ADH3; and plasma HDL cholesterol, apolipoprotein A-I, and apolipoprotein A-II were measured. Nine hundred one subjects had both ADH2 and ADH3 genotypes and HDL cholesterol results, while 753 had both genotypes and all three lipid results. The effect of alcohol intake on the three measured HDL components, and a factor score derived from them, was estimated for each of the ADH2 and ADH3 genotype groups. Results: All the measured components of HDL increased with increasing alcohol consumption over the range of intakes studied, 0-4 drinks per day. There were no significant interactions between alcohol consumption and ADH2 or ADH3 genotypes. Conclusions: The concept that alcohol dehydrogenase genotype and alcohol metabolic rate modify the effects of alcohol on plasma HDL concentration is not supported by our results.

Ozothamnus diosmifolius (Vent.) DC flower and leaf essential oils

Essential oils of rice flower, Ozothamnus diosmifolius, were analyzed by capillary gas chromatograplay-mass spectrometry. Flower oil contained beta-pinene (28.4%) and 1,8-cineole (28.2%), while the leaf oil contained a-pinene (26.0%), beta-pinene (11.6%) and 1,8-cineole (22.2%). Both oils had small amounts of spathulenol (4.1% and 5.2%, respectively).

Codon usage bias and A+T content variation in human papillomavirus genomes

We analyzed the codon usage bias of eight open reading frames (ORFs) across up to 79 human papillomavirus (HPV) genotypes from three distinct phylogenetic groups. All eight ORFs across HPV genotypes show a strong codon usage bias, amongst degenerately encoded amino acids, toward 18 codons mainly with T at the 3rd position. For all 18 degenerately encoded amino acids, codon preferences amongst human and animal PV ORFs are significantly different from those averaged across mammalian genes. Across the HPV types, the L2 ORFs show the highest codon usage bias (73.2 +/- 1.6% and the E4 ORFs the lowest (51.1 +/- 0.5%), reflecting as similar bias in codon 3rd position A + T content (L2: 76.1 +/- 4.2%; E4: 58.6 +/- 4.5%). The E4 ORF, uniquely amongst the HPV ORFs, is G + C rich, while the other ORFs are A + T rich. Codon usage bias correlates positively with A + T content at the codon 3rd position in the E2, E6, L1 and L2 ORFs, but negatively in the E4 ORFs. A general conservation of preferred codon usage across human and non-human PV genotypes whether they originate from a same supergroup or not, together with observed difference between the preferred codon usage for HPV ORFs and for genes of the cells they infect, suggests that specific codon usage bias and A + T content variation may somehow increase the replicational fitness of HPVs in mammalian epithelial cells, and have practical implications for gene therapy of HPV infection. (C) 2003 Elsevier B.V. All rights reserved.

Independent contribution of plaque complexity to myocardial ischemia during Dobutamine stress echocardiography

The influence of complex plaque morphology on the extent of demand-induced ischemia in unselected patients is not well defined. We sought to investigate the functional significance of lesion morphology in patients who underwent coronary angiography and dobutamine stress echocardiography (DSE).,Angiography and DSE were performed within a 6-month period (mean 1 +/- 1 month) in 196 patients. Angiographic assessments involved quantification of stenosis severity, assessment of the extent of jeopardized myocardium, and categorization of plaque morphology according to the Ambrose classification. DSE was interpreted by separate investigators with respect to wall motion score index (WMSI) and number of coronary territories involved. A general linear model was constructed to assess,the independent contribution of patient characteristics and angiographic and DSE results with respect to extent of ischemic myocardium. Complex lesion morphology was seen in 62 patients (32%). Patients with complex lesions were more likely to have had prior myocardial infarction (p < 0.001) and be current smokers (p = 0.03). During angiography, they exhibited a trend toward a greater number of diseased vessels, had a greater coronary jeopardy score (p < 0.001) and more frequent collateral flow (p = 0.03). During echocardiography, patients had a higher stress WMSI (p < 0.001) and were more likely to show ischemia in all 3 arterial territories (p < 0.01). On multivariate regression, the coronary artery jeopardy score and the presence of complex plaque morphology were independent predictors of the extent of ischemic myocardium (R 2 = 34%, p < 0.001). Thus, patients with complex plaque morphology are older, more likely to smoke, and more likely to have had prior myocardial. infarction. They exhibit more extensive disease with higher coronary jeopardy scores and a higher resting and peak stress WMSI. Despite these differences, complex plaque morphology remains an independent predictor of the extent of ischemia during stress. (C) 2003 by Excerpta Medica, Inc.

Relationship of ventricular longitudinal function to contractile reserve in patients with mitral regurgitation

Background Latent left ventricular (LV) dysfunction in patients with valvular or myocardial disease may be identified by loss of contractile reserve (CR) at exercise echocardiography. Contraction in the LV longitudinal axis may be more sensitive than radial contraction to minor disturbances of LV function. We sought to determine whether tissue Doppler measurement of longitudinal function could be used to identify CR. Methods Exercise echocardiography was performed in 86 patients (20 women, age 53 +/- 18 years), 72 with asymptomatic or minimally symptomatic mitral regurgitation, and 14 normal controls. Pulsed-wave tissue Doppler imaging (DTI) was used to measure maximum annular systolic velocity at rest and stress. Inducible ischemia was excluded by analysis of wall motion by an experienced observer. CR was defined by greater than or equal to5% improvement of stress compared with rest ejection fraction (EF). Exercise capacity was assessed from expired gas analysis. Results CR was present in 34 patients with mitral regurgitation (47%); peak EF in patients with and without CR was 74% +/- 11% versus 54% +/- 15% (P

Metodologia para medir a satisfa????o do usu??rio no Canad??: desfazendo mitos e redesenhando roteiros

Caderno composto de relat??rios de pesquisa, elaborados no ??mbito de uma rede de pesquisa formada por altos administradores p??blicos do Canad??, acad??micos e especialistas na ??rea de presta????o de servi??os p??blicos, sobre a avalia????o do cidad??o/usu??rio a respeito da qualidade dos servi??os p??blicos

Exploring the interdisciplinary process in design education

The work presented herein follows an ongoing research that aims to analyze methodological practices to be applied in Design Education. A reflection about methodological strategies in Design Education and the function of drawing in Design represents the beginning of this study. Then, we developed an interdisciplinary pedagogical experience with the Graphic Design 1st grade students from our institution (IPCA). In the current academic year, 2013/2014, we continue to evolve this project, introducing changes in the initial proposal. Major alterations focused on the aspects that could be strengthened in terms of interdisciplinarity. In this article, the authors describe those changes and discuss the outcomes of the novel proposal. As we have already reported, this investigation follows a reflection about working methods to be adopted in Design Education. This is in accordance with other previously published works that purpose the enlargement of Design into new knowledge fields such as Experience or Service Design, changing not only the role of the graphic designer, but also the skills required to be a professional designer (Alain Findelli, 2001), (Brian Lawson, 2006), (Ciampa-Brewer, 2010). Furthermore, concepts such as cooperation or multidisciplinary design, amongst others, have been frequently debated as design teaching strategies (Heller and Talarico, 2011, pp. 82-85). These educational approaches also have an impact on our research. The analysis of all these authors’ contributions together with a reflection on our teaching practice allowed us to propose an improved interdisciplinary intervention.

Efeitos imediatos da mobilização com movimento da rotação medial da tibio-femural m movimento da rot e movimento, em ind com flexão do joelho, na dor e na amplitude de movimento, em indivíduos com síndrome s da mobilização com dor e na amplitude de patelo-femural

Introdução: O síndrome patelo-femural é uma das disfunções músculo-esqueléticas mais comuns ao nível do joelho. É de etiologia multifatorial, sendo a rotação lateral da tíbia um dos fatores contribuintes, sendo que pode potenciar alterações da biomecânica da articulação patelo-femural por aumentar as forças de reação sobre a articulação. Brian Mulligan sugere que a técnica para a correção da rotação lateral da tíbia pode ser benéfica no alívio da dor e no aumento da amplitude de flexão do joelho, em pacientes com síndrome patelo-femural, apesar da evidência acerca da efetividade desta técnica ser ainda escassa. Objetivo: Avaliar os efeitos da técnica de mobilização com movimento de rotação medial da tíbio-femural com flexão do joelho, ao nível da intensidade da dor e da amplitude de movimento de flexão do joelho, durante o agachamento, em indivíduos com síndrome patelo-femural. Métodos: Estudo experimental, com uma amostra constituída por 20 estudantes universitários, do género feminino, com síndrome patelo-femural e dor ao agachamento bilateral. Estes foram distribuídos aleatoriamente por dois grupos: experimental (intervenção com técnica de mobilização com movimento) e placebo (intervenção placebo). Foram avaliadas a amplitude de flexão do joelho com um goniómetro eletrónico (Biometrics®) e a intensidade de dor com a Escala Visual Analógica, durante o agachamento bilateral, antes e imediatamente após as respetivas intervenções. O nível de significância foi de 0,05. Resultados: A realização da Análise da Covariância revelou que, relativamente à intensidade da dor, foi possível constatar que existiram diferenças significativas entre os dois grupos (p<0,001). Entre a avaliação inicial e a final, o grupo experimental diminuiu mais 2,1cm na Escala Visual Analógica do que o grupo placebo. Em relação à avaliação da amplitude articular, foi possível constatar que, existiram diferenças significativas, entre os dois grupos (p=0,004). Entre a avaliação inicial e a final, o grupo experimental teve mais 8,6º de aumento na amplitude articular do que o grupo placebo. Conclusão: Para indivíduos com síndrome patelo-femural, a técnica de mobilização com movimento para correção da rotação lateral da tíbia, parece ser benéfica no alívio da dor e no ganho de amplitude de flexão do joelho, analisando o movimento de agachamento bilateral.