930 resultados para Gated Channel Subunit
Resumo:
Although vaccines have widely been regarded as the most cost-effective way to improve public health, for some organisms new technological advances in vaccine design and delivery, incurring additional developmental costs, will be essential. These organisms are typically those for which natural immunity is either slow to develop or does not develop at all. Clearly, such organisms have evolved strategies to evade immune responses and innovative approaches will be required to induce a type of immune response which is both different to that which develops naturally and is effective. This article describes some approaches to develop vaccines for two such organisms (malaria parasites and Streptococcus pyogenes (group A Streptococcus)) that are associated with widespread mortality and morbidity, mostly in the poorest countries of the world. At this stage, the challenges are primarily scientific, but if these hurdles are surmounted then the challenges will become financial ones - developing much needed vaccines for people least able to afford them. (C) 2002 Australian Society for Parasitology Inc. Published by Elsevier Science Ltd. All rights reserved.
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In renal collecting ducts, a vasopressin-induced cAMP increase results in the phosphorylation of aquaporin-2 (AQP2) water channels at Ser-256 and its redistribution from intracellular vesicles to the apical membrane. Hormones that activate protein kinase C (PKC) proteins counteract this process. To determine the role of the putative kinase sites in the trafficking and hormonal regulation of human AQP2, three putative casein kinase II (Ser-148, Ser-229, Thr-244), one PKC (Ser-231), and one protein kinase A (Ser-256) site were altered to mimic a constitutively non-phosphorylated/phosphorylated state and were expressed in Madin-Darby canine kidney cells. Except for Ser-256 mutants, seven correctly folded AQP2 kinase mutants trafficked as wild-type AQP2 to the apical membrane via forskolin-sensitive intracellular vesicles. With or without forskolin, AQP2-Ser-256A was localized in intracellular vesicles, whereas AQP2-S256D was localized in the apical membrane. Phorbol 12-myristate 13-acetate-induced PKC activation following forskolin treatment resulted in vesicular distribution of all AQP2 kinase mutants, while all were still phosphorylated at Ser-256. Our data indicate that in collecting duct cells, AQP2 trafficking to vasopressin-sensitive vesicles is phosphorylation-independent, that phosphorylation of Ser-256 is necessary and sufficient for expression of AQP2 in the apical membrane, and that PMA-induced PKC-mediated endocytosis of AQP2 is independent of the AQP2 phosphorylation state.
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Compression amplification significantly alters the acoustic speech signal in comparison to linear amplification. The central hypothesis of the present study was that the compression settings of a two-channel aid that best preserved the acoustic properties of speech compared to linear amplification would yield the best perceptual results, and that the compression settings that most altered the acoustic properties of speech compared to linear would yield significantly poorer speech perception. On the basis of initial acoustic analysis of the test stimuli recorded through a hearing aid, two different compression amplification settings were chosen for the perceptual study. Participants were 74 adults with mild to moderate sensorineural hearing impairment. Overall, the speech perception results supported the hypothesis. A further aim of the study was to determine if variation in participants' speech perception with compression amplification (compared to linear amplification) could be explained by the individual characteristics of age, degree of loss, dynamic range, temporal resolution, and frequency selectivity; however, no significant relationships were found.
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There has been much argument about the phylogenetic relationships of the four suborders of lice (Insecta: Phthiraptera). Lyal's study of the morphology of lice indicated that chewing/biting lice (Mallophaga) are paraphyletic with respect to sucking lice (Anoplura). To test this hypothesis we inferred the phylogeny of 33 species of lice from small subunit (SSU) rRNA sequences (18S rRNA). Liposcelis sp. from the Liposcelididae (Psocoptera) was used for outgroup reference. Phylogenetic relationships among the four suborders of lice inferred from these sequences were the same as those inferred from morphology. The Amblycera is apparently the sister-group to all other lice whereas the Rhynchophthirina is apparently sister to the Anoplura; these two suborders are sister to the Ischnocera, i.e. (Amblycera (Ischnocera (Anoplura, Rhynchophthirina))). Thus, the Mallophaga (Amblycera, Ischnocera, Rhynchophthirina) is apparently paraphyletic with respect to the Anoplura. Our analyses also provide evidence that: (i) each of the three suborders of lice that are well represented in our study (the Amblycera, Ischnocera, and Anoplura) are monophyletic; (ii) the Boopiidae is monophyletic; (iii) the genera Heterodoxus and Latumcephalum (Boopiidae) are more closely related to one another than either is to the genus Boopia (also Boopiidae); (iv) the Ricinidae and Laemobothridae may be sister-taxa; (v) the Philopteridae may be paraphyletic with respect to the Trichodectidae; (vi) the genera Pediculus and Pthirus are more closely related to each other than either is to the genus Pedicinus ; and (vii) in contrast to published data for mitochondrial genes, the rates of nucleotide substitution in the SSU rRNA of lice are not higher than those of other insects, nor do substitution rates in the suborders differ substantially from one another.
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Mutations in the E1alpha subunit of the pyruvate dehydrogenase multienzyme complex may result in congenital lactic acidosis, but little is known about the consequences of these mutations at the enzymatic level. Here we characterize two mutants (F205L and T231A) of human pyruvate dehydrogenase in vitro, using the enzyme expressed in Escherichia coli. Wild-type and mutant proteins were purified successfully and their kinetic parameters were measured. F205L shows impaired binding of the thiamin diphosphate cofactor, which may explain why patients carrying this mutation respond to high-dose vitamin B-1 therapy. T231A has very low activity and a greatly elevated K-m for pyruvate, and this combination of effects would be expected to result in severe lactic acidosis. The results lead to a better understanding of the consequences of these mutations on the functional and structural properties of the enzyme, which may lead to improved therapies for patients carrying these mutations.
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delta-Atracotoxin-Ar1a (delta-ACTX-Ar1a) is the major polypeptide neurotoxin isolated from the venom of the male Sydney funnel-web spider, Atrax robustus. This neurotoxin targets both insect and mammalian voltage-gated sodium channels, where it competes with scorpion alpha-toxins for neurotoxin receptor site-3 to slow sodium-channel inactivation. Progress in characterizing the structure and mechanism of action of this toxin has been hampered by the limited supply of pure toxin from natural sources. In this paper, we describe the first successful chemical synthesis and oxidative refolding of the four-disulfide bond containing delta-ACTX-Ar1a. This synthesis involved solid-phase Boc chemistry using double coupling, followed by oxidative folding of purified peptide using a buffer of 2 M GdnHCl and glutathione/glutathiol in a 1:1 mixture of 2-propanol (pH 8.5). Successful oxidation and refolding was confirmed using both chemical and pharmacological characterization. Ion spray mass spectrometry was employed to confirm the molecular weight. H-1 NMR analysis showed identical chemical shifts for native and synthetic toxins, indicating that the synthetic toxin adopts the native fold. Pharmacological studies employing whole-cell patch clamp recordings from rat dorsal root ganglion neurons confirmed that synthetic delta-ACTX-Ar1a produced a slowing of the sodium current inactivation and hyperpolarizing shifts in the voltage-dependence of activation and inactivation similar to native toxin. Under current clamp conditions, we show for the first time that delta-ACTX-Ar1a produces spontaneous repetitive plateau potentials underlying the clinical symptoms seen during envenomation. This successful oxidative refolding of synthetic delta-ACTX-Ar1a paves the way for future structure-activity studies to determine the toxin pharmacophore.
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Venomous animals have evolved a vast array of peptide toxins for prey capture and defence. These peptides are directed against a wide variety of pharmacological targets, making them an invaluable source of ligands for studying the properties of these targets in different experimental paradigms. A number of these peptides have been used in vivo for proof-of-concept studies, with several having undergone preclinical or clinical development for the treatment of pain, diabetes, multiple sclerosis and cardiovascular diseases. Here we survey the pharmacology of venom peptides and assess their therapeutic prospects.
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Introdução - Gated-Single Photon Emission Computed Tomography (Gated-SPECT) do miocárdio usa-se cada vez mais na avaliação conjunta da perfusão do miocárdio e da função ventricular esquerda. O objectivo deste estudo é analisar a possível interferência do número de ciclos cardíacos (ciclos/projecção) e contagens totais por aquisição no cálculo da FEVE (fracção de ejecção do ventrículo esquerdo). Material e métodos - Foram incluídos 35 indivíduos a quem foram realizados estudos Gated-SPECT do miocárdio por indicação clínica. Em todos os pacientes foram adquiridos dois estudos em esforço: um com 25 ciclos! projecção (ES-25) e outro com 50 ciclos! projecção (ES-50), e dois estudos em repouso: um com 25 ciclos/projecção (ER-25) e outro com 12 ciclos! projecção (ER-1 2). Os valores da FEVE obtidos nos diferentes estudos foram tratados estatisticamente com o recurso à versão 14 SPSS. Resultados - Não houve diferenças estatisticamente significativas entre os valores de FEVE ES-25 e ES-50 (p=0.504) nem entre os valores de FEVE ER-25 e ER-12 (p=O.243). Conclusão - No caso da nossa amostra, o número de ciclos!projecção e consequentemente as contagens totais por aquisição não parecem ter influência de forma significativa no cálculo da FEVE%, nem em repouso nem após esforço.
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In this paper a realistic directional channel model that is an extension of the COST 273 channel model is presented. The model uses a cluster of scatterers and visibility region generation based strategy with increased realism, due to the introduction of terrain and clutter information. New approaches for path-loss prediction and line of sight modeling are considered, affecting the cluster path gain model implementation. The new model was implemented using terrain, clutter, street and user mobility information for the city of Lisbon, Portugal. Some of the model's outputs are presented, mainly path loss and small/large-scale fading statistics.
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We report a field-effect phototransistor with a channel comprising a thin nanocrystalline silicon transport layer and a thicker hydrogenated amorphous silicon absorption layer. The semiconductor and dielectric layers were deposited by radio-frequency plasma enhanced chemical vapor deposition. The phototransistor with channel length of 24 microns and photosensitive area of 1.4 mm(2) shows an off-current of about 1 pA, and high photoconductive gain in the subthreshold region. Measurements of the quantum efficiency at different incident light intensities and biasing conditions, along with spectral-response characteristics, and threshold voltage stability characterization demonstrate the feasibility of the phototransistor for low light level detection.
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A imagem de perfusão do miocárdio por tomografia computorizada de emissão de fotão único (SPECT, do acrónimo inglês Single Photon Emission Computed Tomography) é um dos exames complementares de diagnóstico mais indicados na cardiologia clínica. Os dois agentes tecneciados mais utilizados são o 99mTc-tetrofosmina e o 99mTc-sestamibi. Estes permitem avaliar a distribuição do fluxo sanguíneo no músculo cardíaco. O mecanismo de fixação é idêntico nos dois radiofármacos (RF’s). São excretados, do organismo, a partir do sistema hepatobiliar para o duodeno e para o intestino. Assim, o RF está presente tanto no coração como nos órgãos adjacentes. O coração encontra-se localizado sobre o diafragma, logo acima do lobo esquerdo do fígado e nas proximidades do estômago. Deste modo, os fotões difusos provenientes desses órgãos podem interferir com a interpretação das imagens SPECT, principalmente na parede inferior do ventrículo esquerdo, traduzindo-se na redução da razão alvo-fundo e na qualidade da imagem. Deste modo, a sensibilidade e a especificidade da Cintigrafia de Perfusão do Miocárdio (CPM) diminuem. Têm sido descritos na literatura vários métodos e técnicas para minimizar o efeito da actividade extra-miocárdica, como: a aquisição de imagens mais tardias, a ingestão de líquidos ou alimentos ricos em lípidos. Outras técnicas apontam para a alteração do posicionamento do paciente durante a aquisição. Assim, o objectivo deste estudo é identificar o protocolo da CPM a nível nacional e avaliar a percentagem de repetições de exames provocadas pela interferência de actividade extra-miocárdica.
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A crucial method for investigating patients with coronary artery disease (CAD) is the calculation of the left ventricular ejection fraction (LVEF). It is, consequently, imperative to precisely estimate the value of LVEF--a process that can be done with myocardial perfusion scintigraphy. Therefore, the present study aimed to establish and compare the estimation performance of the quantitative parameters of the reconstruction methods filtered backprojection (FBP) and ordered-subset expectation maximization (OSEM). Methods: A beating-heart phantom with known values of end-diastolic volume, end-systolic volume, and LVEF was used. Quantitative gated SPECT/quantitative perfusion SPECT software was used to obtain these quantitative parameters in a semiautomatic mode. The Butterworth filter was used in FBP, with the cutoff frequencies between 0.2 and 0.8 cycles per pixel combined with the orders of 5, 10, 15, and 20. Sixty-three reconstructions were performed using 2, 4, 6, 8, 10, 12, and 16 OSEM subsets, combined with several iterations: 2, 4, 6, 8, 10, 12, 16, 32, and 64. Results: With FBP, the values of end-diastolic, end-systolic, and the stroke volumes rise as the cutoff frequency increases, whereas the value of LVEF diminishes. This same pattern is verified with the OSEM reconstruction. However, with OSEM there is a more precise estimation of the quantitative parameters, especially with the combinations 2 iterations × 10 subsets and 2 iterations × 12 subsets. Conclusion: The OSEM reconstruction presents better estimations of the quantitative parameters than does FBP. This study recommends the use of 2 iterations with 10 or 12 subsets for OSEM and a cutoff frequency of 0.5 cycles per pixel with the orders 5, 10, or 15 for FBP as the best estimations for the left ventricular volumes and ejection fraction quantification in myocardial perfusion scintigraphy.
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Myocardial perfusion-gated-SPECT (MP-gated-SPECT) imaging often shows radiotracer uptake in abdominal organs. This accumulation interferes frequently with qualitative and quantitative assessment of the infero-septal region of myocardium. The objective of this study is to evaluate the effect of ingestion of different fat content on the reduction of extra-myocardial uptake and to improve MP-gated-SPECT image quality. In this study, 150 patients (65 ^ 18 years) who were referred for MP-gated-SPECT underwent a 1-day-protocol including imaging after stress (physical or pharmacological) and resting conditions. All patients gave written informed consent. Patients were subdivided into five groups: GI, GII, GIII, GIV and GV. In the first four groups, patients ate two chocolate bars with different fat content. Patients in GV – control group (CG) – had just water. Uptake indices (UI) of myocardium (M)/liver(L) and M/stomach–proximal bowel(S) revealed lower UI of M/S at rest in all groups. Both stress and rest studies using different food intake indicate that patients who ate chocolate with different fat content showed better UI of M/L than the CG. The UI of M/L and M/S of groups obtained under physical stress are clearly superior to that of groups obtained under pharmacological stress. These differences are only significant in patients who ate high-fat chocolate or drank water. The analysis of all stress studies together (GI, GII, GIII and GIV) in comparison with CG shows higher mean ranks of UI of M/L for those who ate high-fat chocolate. After pharmacological stress, the mean ranks of UI of M/L were higher for patients who ate high- and low-fat chocolate. In conclusion, eating food with fat content after radiotracer injection increases, respectively, the UI of M/L after stress and rest in MP-gated-SPECT studies. It is, therefore, recommended that patients eat a chocolate bar after radiotracer injection and before image acquisition.
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Myocardial Perfusion Gated Single Photon Emission Tomography (Gated-SPET) imaging is used for the combined evaluation of myocardial perfusion and left ventricular (LV) function. But standard protocols of the Gated-SPECT studies require long acquisition times for each study. It is therefore important to reduce as much as possible the total duration of image acquisition. However, it is known that this reduction leads to decrease on counts statistics per projection and raises doubts about the validity of the functional parameters determined by Gated-SPECT. Considering that, it’s difficult to carry out this analysis in real patients. For ethical, logistical and economical matters, simulated studies could be required for this analysis. Objective: Evaluate the influence of the total number of counts acquired from myocardium, in the calculation of myocardial functional parameters (LVEF – left ventricular ejection fraction, EDV – end-diastolic volume, ESV – end-sistolic volume) using routine software procedures.
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Introdução – A cintigrafia de perfusão do miocárdio (CPM) é um dos exames complementares de diagnóstico mais indicados na patologia cardiovascular. A atividade extramiocárdica é um problema recorrente na aquisição e avaliação da imagem. Parâmetros como o radiofármaco (RF) administrado, o tipo de esforço, a ingestão de alimentos, o posicionamento e o processamento de imagem influenciam a atividade extramiocárdica na CPM. O objetivo deste estudo é identificar o protocolo da CPM a nível nacional e avaliar a percentagem de repetições das aquisições provocadas por este problema. Metodologia – Para a realização deste estudo, a população selecionada foram os Serviços de Medicina Nuclear (SMN) em Portugal Continental. Foram analisados 15 questionários, respondidos pelos Técnicos Coordenadores dos vários SMN. Foi realizada a associação do comportamento das variáveis, através da estatística descritiva e do Coeficiente de Correlação de Spearman. Resultados – A percentagem de repetições é menor com o uso da 99mTecnécio(Tc)-Tetrofosmina, aquisição de imagens tardias, ingestão de diversos alimentos (água, leite, sumo, pão com manteiga) e com a correção de atenuação. Conclusão – Os resultados obtidos demonstram que a maioria das soluções aplicadas nos SMN resultam numa diminuição das repetições.