Tubular system volume changes in twitch fibres from toad and rat skeletal muscle assessed by confocal microscopy

The volume of the extracellular compartment (tubular system) within intact muscle fibres from cane toad and rat was measured under various conditions using confocal microscopy. Under physiological conditions at rest, the fractional volume of the tubular system (t-sys(Vol)) was 1.38 +/- 0.09% (n = 17),1.41 +/- 0.09% (n = 12) and 0.83 +/- 0.07% (n = 12) of the total fibre volume in the twitch fibres from toad iliofibularis muscle, rat extensor digitorum longus muscle and rat soleus muscle, respectively. In toad muscle fibres, the t-sys(Vol) decreased by 30% when the tubular system was fully depolarized and decreased by 15% when membrane cholesterol was depleted from the tubular system with methyl-beta-cyclodextrin but did not change as the sarcomere length was changed from 1.93 to 3.30 mum. There was also an increase by 30% and a decrease by 25% in t-sys(Vol) when toad fibres were equilibrated in solutions that were 2.5-fold hypertonic and 50% hypotonic, respectively. When the changes in total fibre volume were taken into consideration, the t-sys(Vol) expressed as a percentage of the isotonic fibre volume did actually decrease as tonicity increased, revealing that the tubular system in intact fibres cannot be compressed below 0.9% of the isotonic fibre volume. The results can be explained in terms of forces acting at the level of the tubular wall. These observations have important physiological implications showing that the tubular system is a dynamic membrane structure capable of changing its volume in response to the membrane potential, cholesterol depletion and osmotic stress but not when the sarcomere length is changed in resting muscle.

Properties of the vertebrate skeletal muscle tubular system as a sealed compartment

Confocal imaging of impermeant fluorescent dyes trapped in the tubular (t-) system of skeletal muscle fibres of rat and cane toad was used to examine changes in the morphology of the t-system upon mechanical skinning, the time course of dye loss from the sealed t-systern in mechanically skinned fibres and the influence of rapid application and removal of glycerol on the morphology of the sealed t-system. In contrast to intact fibres, which have a t-systern open to the outside, the sealed t-systern of toad mechanically skinned fibres consistently displayed local swellings (vesicles). The occurrence of vesicles in the sealed t-system of rat-skinned fibres was infrequent. Application and removal of 200-400 mM glycerol to the sealed t-system did not produce any obvious changes in its morphology. The dyes fluo-3, fura-2 and Oregon green 488 were lost from the sealed t-system of toad fibres at different rates suggesting that the mechanism of organic anion transport across the tubular wall was not by indiscriminate bulk transport. The rate of fluo-3 and fura-2 loss from the sealed t-system of rat fibres was greater in rat than in toad fibres and could be explained by differences in surface area: volume ratio of the t-system in the two fibre types. Based on the results presented here and on other results from this laboratory, an explanation is given for the formation of numerous vesicles in toad-skinned fibres and lack of vesicle formation in rat-skinned fibres. This explanation can also help with better understanding the mechanism responsible for vacuole formation in intact fibres. (C) 2002 Elsevier Science Ltd. All rights reserved.

Osmotic properties of the sealed tubular system of toad and rat skeletal muscle

A method was developed that allows conversion of changes in maximum Ca2+-dependent fluorescence of a fixed amount of fluo-3 into volume changes of the fluo-3-containing solution. This method was then applied to investigate by confocal microscopy the osmotic properties of the sealed tubular (t-) system of toad and rat mechanically skinned fibers in which a certain amount Of fluo-3 was trapped. When the osmolality of the myoplasmic environment was altered by simple dilution or addition of sucrose within the range 190-638 mosmol kg(-1), the sealed t-system of toad fibers behaved almost like an ideal osmometer, changing its volume inverse proportionally to osmolality However, increasing the osmolality above 638 to 2,550 mosmol kg(-1) caused hardly any change in t-system volume. In myoplasmic solutions made hypotonic to 128 mosmol kg(-1), a loss of Ca2+ from the sealed t-system of toad fibers Occurred, presumably through either stretch-activated cationic channels or store-operated Ca2+ channels. In contrast to the behavior of the t-system in toad fibers, the volume of the sealed t-system of rat fibers changed little (by

Histological features of acute tubular necrosis in native kidneys and long-term renal function

There are few studies on the relationship between the morphology of acute tubular necrosis (ATN) in native kidneys and late functional recovery. Eighteen patients with acute renal failure (ARF) who had undergone renal biopsy were studied. All had the histological diagnosis of ATN and were followed for at least six months. Clinical characteristics of ARF were analyzed, and histological features were semi-quantitatively evaluated (tubular atrophy, interstitial inflammatory infiltrate, interstitial fibrosis, and ATN). According to the maximal GFR achieved during the follow-up, patients were divided into two groups: complete recovery (GFR >= 90 mL/min/1.73 m(2)) and partial recovery (GFR < 90 mL/min/1.73 m(2)). Only 39% of the patients achieved complete recovery. Patients with partial recovery achieved their maximal GFR (63 +/- 9 mL/min/1.73 m(2)) 37 +/- 14 months after ARF, a period of time similar to those patients with complete recovery (i.e., 54 +/- 22 months). Patients with partial recovery had more severe ARF: oliguria was more frequent (90 versus 17%, p < 0.01), and they had higher peak creatinine (13.85 +/- 1.12 versus 8.95 +/- 1.30 mg/dL, p = 0.01), and longer hospitalization (45 +/- 7 versus 20 +/- 4 days, p = 0.03). No single histological parameter was associated with partial recovery, but the sum of all was when expressed as an injury index [4.00 (2.73-5.45) versus 2.00 (1.25-3.31), p < 0.05]. In conclusion, among patients with atypical ATN course, those with more severe ARF and tubule-interstitial lesions are more prone to partial recovery.

Tubular eyes of deep-sea fishes: A comparative study of retinal topography

The world's deep oceans are home to a number of teleosts with asymmetrical or tubular eyes. These immobile eyes possess large spherical lenses and subtend a large binocular visual field directed either dorsally or rostrally. Derived from a lateral non-tubular eye, the tubular eye is comprised of a thick main retina, subserving the rostrally or dorsally directed binocular visual field, and a thin accessory retina subserving, the lateral, monocular visual field. The main retina is thought to receive a focussed image, while the accessory retina is too close to the lens for a focussed image to be received. Several species also possess retinal diverticula, which are small evaginations of differentiated retina located in the rostrolateral wall of the eye and thought to increase the visual field. In order to investigate the spatial resolving power of these retinae (main, accessory and diverticulum), the distribution of cells within the ganglion cell layer was analysed from retinal wholemounts and sectioned material in ten species representing four genera. In all species, the main retina possesses a marked increase in cell density towards a specialised retinal region (area centralis), with a centro-peripheral gradient range between 7.1 and 60:1 and a peak density range of between 30 and 55 x 10(3) cells per mm(2). The accessory retinae and the transitional zone between the main and accessory retinae possess relatively low cell densities (between 1 and 10 x 10(3) cells per mm(2)) and lack an area centralis. Retinal diverticula examined in four species possess mean ganglion cell densities of between 7.2 and 109.4 x 10(3) cells per mm(2). Analyses of soma areas show that the ganglion cell layer of most species possesses cells with areas in a range of 8.0 to 15.4 mu m(2) in the main retina and between 15.1 and 17.4 mu m(2) in the accessory retina. The peak spatial resolving power of the main retina of the ten species varies from 4.1 to 9.1 cycles per degree. The positions of the retinal areae centrales relative to each species' binocular visual field are discussed in relation to what is known of feeding behaviour of these fishes in the deep-sea.

Role of p21 and oxidative stress on renal tubular resistance after acute ischaemic injury

Background. Subsequent ischaemic episodes may induce renal resistance. P21 is a cell cycle inhibitor that may be induced by oxygen-free radicals and may have a protective effect in ischaemic acute kidney injury (AKI). This study aimed at evaluating the role of oxidative stress and p21 on tubular resistance in a model of acquired resistance after renal ischaemia and in isolated renal tubules. Methods. Wistar rats were divided into: Group 1-sham; Group 2-sham operated and after 2 days submitted to 45-min ischaemia; and Group 3-45-min ischaemia followed after 2 days by a second 45-min ischaemia. Plasma urea was evaluated on Days 0, 2 and 4. Serum creatinine, creatinine clearance and oxidants (thiobarbituric acid-reactive substances) were determined 48 h after the second procedure (Day 4). Histology, immunohistochemistry for lymphocytes (CD3), macrophages (ED1), proliferation (PCNA) and apoptosis (TUNEL) were also evaluated. Rat proximal tubules (PTs) were isolated by collagenase digestion and Percoll gradient from control rats and rats previously subjected to 35 min of ischaemia. PTs were submitted to 15-min hypoxia followed by 45-min reoxygenation. Cell injury was assessed by lactate dehydrogenase release and hydroperoxide production (xylenol orange). Results. Ischaemia induced AKI in Group 2 and 3 rats. Subsequent ischaemia did not aggravate renal injury, demonstrating renal resistance (Group 3). Renal function recovery was similar in Group 2 and 3. Plasma and urine oxidants were similar among in Group 2 and 3. Histology disclosed acute tubular necrosis in Group 2 and 3. Lymphocyte infiltrates were similar among all groups whereas macrophages infiltrate was greater in Group 3. Cell proliferation was greater in Group 2 compared with Group 3. Apoptosis was similar in groups 2 and 3. The p21 expression was increased only in Group 3 whereas it was similar in groups 1 and 2. PTs from the ischaemia group were sensitive to hypoxia but resistant to reoxygenation injury which was followed by lower hydroperoxide production compared to control PT. Conclusion. Renal resistance induced by ischaemia was associated with cell mechanism mediators involving oxidative stress and increased p21 expression.

Bcl-X-L translocation in renal tubular epithelial cells in vitro protects distal cells from oxidative stress

Background. The molecular pathogenesis of different sensitivities of the renal proximal and distal tubular cell populations to ischemic injury, including ischemia-reperfusion (IR)-induced oxidative stress, is not well-defined. An in vitro model of oxidative stress was used to compare the survival of distal [Madin-Darby canine kidney (MDCK)] and proximal [human kidney-2 (HK-2)] renal tubular epithelial cells, and to analyze for links between induced cell death and expression and localization of selected members of the Bcl-2 gene family (anti-apoptotic Bcl-2 and Bcl-X-L, pro-apoptotic Bax and Bad), Methods. Cells were treated with 1 mmol/L hydrogen peroxide (H2O2) Or were grown in control medium for 24 hours. Cell death (apoptosis) was quantitated using defined morphological criteria. DNA gel electrophoresis was used for biochemical identification. Protein expression levels and cellular localization of the selected Bcl-2 family proteins were analyzed (West ern immunoblots, densitometry, immunoelectron microscopy). Results. Apoptosis was minimal in control cultures and was greatest in treated proximal cell cultures (16.93 +/- 4.18% apoptosis) compared with treated distal cell cultures (2.28 +/- 0.85% apoptosis, P < 0.001). Endogenous expression of Bcl-X-L and Bax, but not Bcl-2 or Bad, was identified in control distal cells, Bcl-X-L and Bax had nonsignificant increases (P > 0.05) in these cells. Bcl-2, Bax, and Bcl-X-L, but not Bad, were endogenously expressed in control proximal cells. Bcl-X-L was significantly decreased in treated proximal cultures (P < 0.05), with Bas and Bcl-2 having nonsignificant increases (P > 0.05). Immunoelectron microscopy localization indicated that control and treated hut surviving proximal cells had similar cytosolic and membrane localization of the Bcl-2 proteins. In comparison, surviving cells in the treated distal cultures showed translocation of Bcl-X-L from cytosol to the mitochondria after treatment with H2O2, a result that was confirmed using cell fractionation and analysis of Bcl-XL expression levels of the membrane and cytosol proteins. Bax remained distributed evenly throughout the surviving distal cells, without particular attachment to any cellular organelle. Conclusion. The results indicate that in this in vitro model, the increased survival of distal compared with proximal tubular cells after oxidative stress is best explained by the decreased expression of anti-apoptotic Bcl-X-L in proximal cells, as well as translocation of Bcl-X-L protein to mitochondria within the surviving distal cells.

Theoretical and experimental studies of unbraced tubular trusses allowing for torsional stiffness

This paper describes the buckling phenomenon of a tubular truss with unsupported length through a full-scale test and presents a practical computational method for the design of the trusses allowing for the contribution of torsional stiffness against buckling, of which the effect has never been considered previously by others. The current practice for the design of a planar truss has largely been based on the linear elastic approach which cannot allow for the contribution of torsional stiffness and tension members in a structural system against buckling. The over-simplified analytical technique is unable to provide a realistic and an economical design to a structure. In this paper the stability theory is applied to the second-order analysis and design of the structural form, with detailed allowance for the instability and second-order effects in compliance with design code requirements. Finally, the paper demonstrates the application of the proposed method to the stability design of a commonly adopted truss system used in support of glass panels in which lateral bracing members are highly undesirable for economical and aesthetic reasons.

Efeito da geometria dos dentes da serra de fita na produção de madeira serrada de eucalipto

O objetivo deste trabalho foi determinar o efeito dos parâmetros que compõem os dentes de serra de fita na produção e qualidade da madeira serrada de eucalipto. Analisou-se o efeito de dois passos de dentes (57,15 mm e passo variado: 50,8 - 50,8 - 50,39 - 50,71 - 60,03 - 60,35 mm), de dois ângulos de ataque (26 e 270) e de duas alturas (22 e 25,4 mm) em duas classes diamétricas (25 a 34 cm e 35 e 47 cm) de clones de híbridos Eucalyptus grandis x Eucalyptus urophylla com 15 anos de idade. Não houve ganho de qualidade quando se trabalhou com passo variado. As lâminas de serra que possuíam ângulo de ataque de 270 e passo único tanto com altura do dente de 22 mm quanto com 25,4 mm geraram desvios acima da espessura-meta. A menor variação em torno da espessura-meta foi obtida quando se utilizaram lâminas de serra de ângulo de ataque de 270 e altura do dente de 25,4 mm. O maior desvio ocorreu quando o desdobro foi efetuado com passo variado e ângulo de ataque de 260. A utilização de passo variado, de modo geral, foi o que apresentou maiores desvios na espessura das tábuas em relação à espessura desejada.

A geometria na formação inicial de professores: contributos para a caracterização dos estudantes

Dissertação apresentada para obtenção do grau de Mestre em Educação Matemática na Educação Pré-Escolar e nos 1.º e 2.º Ciclos do Ensino Básico

Análise estrutural de uma torre tubular sob acção do vento

Com a realização deste trabalho, pretende-se essencialmente dar a conhecer a influência que a acção do vento possui no dimensionamento de determinadas estruturas, neste caso específico, em torres tubulares de telecomunicações. É por todos sabido, da importância de um capaz e evoluído sistema de comunicações, no desenvolvimento sustentado do mundo moderno. Nesse sentido, o avanço galopante, principalmente nas últimas duas décadas, das tecnologias de Telecomunicações, implicou uma rápida resposta em consonância na implantação e proliferação de infraestruturas de suporte aos equipamentos dessas tecnologias. Assim, a estrutura em forma de torre tubular, entre as demais variadas secções que as constituem, foi adquirindo preponderância neste campo, constituindo a mais vasta utilização de estruturas de suporte aos equipamentos de telecomunicações, nomeadamente em meios rurais, onde escasseiam edificações com alturas suficientes para fazer face às necessidades das operadoras licenciadas para os devidos efeitos. Será efectuada uma breve descrição sobre as diversas e mais comuns tipologias de torres utilizadas no âmbito do suporte de equipamentos de telecomunicações. Descreve-se, em forma de relatório e levantamento fotográfico, a ocorrência do colapso de uma torre tubular de telecomunicações. Por último, e na sequência do incidente referido no ponto anterior, será efectuada com detalhe, a análise estrutural da torre tubular que foi instalada na posição da anteriormente instalada.

Os alunos surdos e a matemática: um projeto de intervenção em geometria

O presente estudo é um projeto de investigação-ação, que contou com a participação de três alunos surdos do 8º ano, de uma Escola Pública de Lisboa, que se assume vocacionada para o ensino de surdos. Os alunos referidos possuem diferentes graus de surdez e diferentes formas de comunicação. Neste trabalho, procurámos: (i) conhecer as estratégias utilizadas pelos alunos para a resolução de problemas de geometria; (ii) conhecer as formas de comunicação entre professor e alunos para promover a resolução de problemas de geometria, nomeadamente na abordagem do teorema de Pitágoras; (iii) contribuir para o desenvolvimento de competências específicas para a compreensão e resolução de problemas de geometria; (iv) reforçar a noção da utilidade da geometria na vida quotidiana. Após a caracterização inicial da situação pedagógica, elaborámos um plano de de intervenção que implementámos e monitorizámos, procedendo no final à avaliação dos resultados. Os dados foram recolhidos através da observação participante das aulas de Matemática, das entrevistas realizadas aos alunos, das conversas informais e da análise de diversos documentos. A análise dos dados recolhidos durante o processo de intervenção permitiu identificar as formas de comunicação e estratégias de ensino mais utilizadas pela professora, bem como as formas de comunicação e as estratégias de aprendizagem que os alunos usam. A análise dos resultados mostra que os alunos desenvolveram capacidades ao nível da compreensão do conceito de forma das figuras geométricas e da resolução de problemas geométricos através de construções, embora nem todos tenham atingido o mesmo nível.

Contributos de um ambiente de geometria dinâmica (geogebra) e do geoplano na compreensão das propriedades e relações entre quadriláteros: um estudo com alunos do 4.º ano

O estudo insere-se no âmbito da educação matemática, mais especificamente na área da geometria. Com esta investigação pretende-se compreender qual o contributo do Ambiente de Geometria Dinâmica (AGD), GeoGebra, e do material manipulável (geoplano) na identificação das propriedades e relações entre quadriláteros: trapézio, paralelogramo, retângulo, losango e quadrado. De acordo com o objetivo do estudo formularam-se três questões: Qual o papel das representações na identificação das propriedades dos quadriláteros? Que influência tem a visualização na identificação das propriedades dos quadriláteros? Quais os contributos da utilização do AGD, GeoGebra, e do material manipulável (geoplano) na compreensão das propriedades e relações entre os quadriláteros? Atendendo à problemática em estudo, optou-se por uma metodologia de investigação predominantemente qualitativa, baseada em três estudos de caso. A recolha de dados empíricos foi realizada numa turma de 4.º ano do 1.º ciclo do ensino básico onde se implementou uma experiência de ensino, envolvendo todos os alunos e recorrendo ao geoplano e ao GeoGebra. Essa recolha incidiu na observação participante, em sala de aula, no registo de notas da investigadora, nos documentos produzidos pelos alunos e nos registos áudio e vídeo das discussões nos grupos e no coletivo da turma. Em termos de resultados, salienta-se que a utilização de representações evidenciou a compreensão que os alunos têm dos conceitos, facilitou a identificação das propriedades dos quadriláteros e a compreensão das relações entre eles. Os alunos identificaram as propriedades com base nas representações, no entanto focaram-se em casos particulares, de acordo com a imagem mental que têm da figura, especificamente o protótipo, indiciando a influência da visualização. Dos resultados sobressai também a dificuldade que os alunos sentiram em considerar uma figura como representante de uma classe e em distinguir atributos essenciais e não essenciais. Da análise dos dados ressalta também que tanto o geoplano como o GeoGebra foram uma mais-valia na concretização da experiência de ensino deste estudo. Os dois possuem vantagens e limitações e podem ser usados de maneira que um complemente o outro, podendo dessa forma contribuir, favoravelmente, para a aprendizagem da geometria. - ABSTRACT The study falls within the scope of mathematics education, specifically in the area of geometry. This research aims to understand the contribution of the Dynamic Geometry Environment (GeoGebra) and manipulative material (Geoboard) in the identification of properties and relations of quadrilaterals: trapezium, parallelogram, rectangle, diamond and square. According to the purpose of the study three questions were formulated: What is the role of the representations in the identification of the properties of quadrilaterals? What is the influence of visualization on the identification of the properties of quadrilaterals? What are the contributions on the use of Dynamic Geometry Environment (GeoGebra) and manipulative material (Geoboard) to understand the properties and relationships among quadrilaterals? Given the problem under study, it was chosen a predominantly qualitative research methodology based on three case studies. The empirical data collection was carried out on a fourth primary grade class, where it was implemented a teaching experience, involving all students and the use of Geoboard and GeoGebra. This collection was focused on participant observation in the classroom, in the record of the researcher’s notes, the documents produced by the students and the audio and video recordings of group discussions and collective class. In terms of results, it is noted that the use of representations showed the students understanding of the concepts and that it has facilitated the identification of the properties of quadrilaterals and understanding of the relationships between them. The students identified properties based on representations, however they focused in particular cases, according to the mental image that they have of the figure, specifically the prototype, indicating the influence of visualization. Results also highlights the difficulty that students felt in considering a figure as a representation of a group and distinguishing essential and nonessential attributes. Data analysis also highlights that both Geoboard and GeoGebra have been an asset in the delivery of teaching experience in this study. Both have advantages and limitations and they may be used to complement each other and contribute favorably to the learning of geometry.