Carpenter Syndrome: Extended RAB23 Mutation Spectrum and Analysis of Nonsense-mediated mRNA Decay

Carpenter syndrome, a rare autosomal recessive disorder characterized by a combination of craniosynostosis, polysyndactyly, obesity, and other congenital malformations, is caused by mutations in RAB23, encoding a member of the Rab-family of small GTPases. In 15 out of 16 families previously reported, the disease was caused by homozygosity for truncating mutations, and currently only a single missense mutation has been identified in a compound heterozygote. Here, we describe a further 8 independent families comprising 10 affected individuals with Carpenter syndrome, who were positive for mutations in RAB23. We report the first homozygous missense mutation and in-frame deletion, highlighting key residues for RAB23 function, as well as the first splice-site mutation. Multi-suture craniosynostosis and polysyndactyly have been present in all patients described to date, and abnormal external genitalia have been universal in boys. High birth weight was not evident in the current group of patients, but further evidence for laterality defects is reported. No genotype-phenotype correlations are apparent. We provide experimental evidence that transcripts encoding truncating mutations are subject to nonsense-mediated decay, and that this plays an important role in the pathogenesis of many RAB23 mutations. These observations refine the phenotypic spectrum of Carpenter syndrome and offer new insights into molecular pathogenesis. (C) 2011 Wiley-Liss, Inc.

Expression of Circadian Clock and Melatonin Receptors within Cultured Rat Cardiomyocytes

Melatonin, the pineal gland hormone, provides entrainment of many circadian rhythms to the ambient light/dark cycle. Recently, cardiovascular studies have demostrated melatonin interactions with many physiological processes and diseases, such as hypertension and cardiopathologies. Although membrane melatonin receptors (MT1, MT2) and the transcriptional factor ROR alpha have been reported to be expressed in the heart, there is no evidence of the cell-type expressing receptors as well as the possible role of melatonin on the expression of the circadian clock of cardiomyocytes, which play an important role in cardiac metabolism and function. Therefore, the aim of this study was to evaluate the mRNA and protein expressions of MT1, MT2, and ROR alpha and to determine whether melatonin directly influences expression of circadian clocks within cultured rat cardiomyocytes. Adult rat cardiomyocyte cultures were created, and the cells were stimulated with 1 nM melatonin or vehicle. Gene expressions were assayed by real-time polymerase chain reaction (PCR). The mRNA and protein expressions of membrane melatonin receptors and RORa were established within adult rat cardiomyocytes. Two hours of melatonin stimulation did not alter the expression pattern of the analyzed genes. However, given at the proper time, melatonin kept Rev-erb alpha expression chronically high, specifically 12 h after melatonin treatment, avoiding the rhythmic decline of Rev-erb alpha mRNA. The blockage of MT1 and MT2 by luzindole did not alter the observed melatonin-induced expression of Rev-erb alpha mRNA, suggesting the nonparticipation of MT1 and MT2 on the melatonin effect within cardiomyocytes. It is possible to speculate that melatonin, in adult rat cardiomyocytes, may play an important role in the light signal transduction to peripheral organs, such as the heart, modulating its intrinsic rhythmicity. (Author correspondence: cipolla@icb.usp.br)

Exercise Training Reduces Insulin Resistance and Upregulates the mTOR/p70S6k Pathway in Cardiac Muscle of Diet-Induced Obesity Rats

Obesity and insulin resistance are rapidly expanding public health problems. These disturbances are related to many diseases, including heart pathology. Acting through the Akt/mTOR pathway, insulin has numerous and important physiological functions, such as the induction of growth and survival of many cell types and cardiac hypertrophy. However, obesity and insulin resistance can alter mTOR/p70S6k. Exercise training is known to induce this pathway, but never in the heart of diet-induced obesity subjects. To evaluate the effect of exercise training on mTOR/p70S6k in the heart of obese Wistar rats, we analyzed the effects of 12 weeks of swimming on obese rats, induced by a high-fat diet. Exercise training reduced epididymal fat, fasting serum insulin and plasma glucose disappearance. Western blot analyses showed that exercise training increased the ability of insulin to phosphorylate intracellular molecules such as Akt (2.3-fold) and Foxo1 (1.7-fold). Moreover, reduced activities and expressions of proteins, induced by the high-fat diet in rats, such as phospho-JNK (1.9-fold), NF-kB (1.6-fold) and PTP-1B (1.5-fold), were observed. Finally, exercise training increased the activities of the transduction pathways of insulin-dependent protein synthesis, as shown by increases in Raptor phosphorylation (1.7-fold), p70S6k phosphorylation (1.9-fold), and 4E-BP1 phosphorylation (1.4-fold) and a reduction in atrogin-1 expression (2.1-fold). Results demonstrate a pivotal regulatory role of exercise training on the Akt/ mTOR pathway, in turn, promoting protein synthesis and antagonizing protein degradation. J. Cell. Physiol. 226: 666-674, 2011. (C) 2010 Wiley-Liss, Inc.

A statistical evaluation of the field emission for copper oxide nanostructures

A statistical data analysis methodology was developed to evaluate the field emission properties of many samples of copper oxide nanostructured field emitters. This analysis was largely done in terms of Seppen-Katamuki (SK) charts, field strength and emission current. Some physical and mathematical models were derived to describe the effect of small electric field perturbations in the Fowler-Nordheim (F-N) equation, and then to explain the trend of the data represented in the SK charts. The field enhancement factor and the emission area parameters showed to be very sensitive to variations in the electric field for most of the samples. We have found that the anode-cathode distance is critical in the field emission characterization of samples having a non-rigid nanostructure. (C) 2007 Elsevier B.V. All rights reserved.

The concept of quasi-integrability: a concrete example

We use the deformed sine-Gordon models recently presented by Bazeia et al [1] to take the first steps towards defining the concept of quasi-integrability. We consider one such definition and use it to calculate an infinite number of quasi-conserved quantities through a modification of the usual techniques of integrable field theories. Performing an expansion around the sine-Gordon theory we are able to evaluate the charges and the anomalies of their conservation laws in a perturbative power series in a small parameter which describes the ""closeness"" to the integrable sine-Gordon model. We show that in the case of the two-soliton scattering the charges, up to first order of perturbation, are conserved asymptotically, i.e. their values are the same in the distant past and future, when the solitons are well separated. We indicate that this property may hold or not to higher orders depending on the behavior of the two-soliton solution under a special parity transformation. For closely bound systems, such as breather-like field configurations, the situation however is more complex and perhaps the anomalies have a different structure implying that the concept of quasi-integrability does not apply in the same way as in the scattering of solitons. We back up our results with the data of many numerical simulations which also demonstrate the existence of long lived breather-like and wobble-like states in these models.

Constraining the age of the iporanga formation with SHRIMP U-Pb zircon: Implications for possible ediacaran glaciation in the Ribeira Belt, SE Brazil

The Ribeira belt in SE Brazil is a Neoproterozoic to Early Palaeozoic orogen, whose architecture and history is not yet fully understood. The depositional age of many of the sedimentary sequences in the Ribeira Belt remains unconstrained, and with debate concerning their depositional environment and tectonic setting. In this paper we present SHRIMP zircon U/Pb age constraints for one such problematic unit in the Ribeira Belt the lporanga Formation - and discuss the significance of this age with regards to the timing of Neoproterozoic glacial events in southeast Brazil. Using a felsic volcanic unit immediately under the lporanga Formation and granite cobbles from breccias in its basal parts a reconnaissance SHRIMP U/Pb zircon maximum depositional age of 580 Ma is assigned for the base of this unit. This age is marginally younger than the 625605 Ma ages for intrusions into the Lajeado and Ribeira subgroups, with which the lporanga Formation is in tectonic contact. This indicates that the Lajeado and Ribeira subgroups are not stratigraphically equivalent to the lporanga Formation, as thought previously by some workers. The maximum depositional age of 580 Ma also places a maximum time constraint on the tectonic juxtaposition of the lporanga Formation with other supracrustal units, and on the greenschist facies metamorphism and isoclinal folding that affected it. The potential glacial origin for the lporanga Formation, if correct, would place it in the late Ediacaran - provisionally equivalent to the Gaskiers glaciation. (c) 2007 International Association for Gondwana Research. Published by Elsevier B.V. All rights reserved.

Solvatochromism in Binary Mixtures: First Report on a Solvation Free Energy Relationship between Solvent Exchange Equilibrium Constants and the Properties of the Medium

We have employed UV-vis spectroscopy in order to investigate details of the solvation of six solvatochromic indicators, hereafter designated as ""probes"", namely, 2,6-diphenyl-4-(2,4,6-triphenylpyridinium-1-yl) phenolate (RB); 4-[(E)-2-(1-methylpyridinium-4-yl)ethenyl] phenolate, MePM; 1-methylquinolinium-8-olate, QB; 2-bromo-4-[(E)-2-(1-methylpyridinium-4-yl)ethenyl] phenolate, MePMBr, 2,6-dichloro-4-(2,4,6-triphenylpyridinium-1-yl) phenolate (WB); and 2,6-dibromo-4-[(E)-2-(1-methylpyridinium-4-yl)ethenyl] phenolate, MePMBr,, respectively. These can be divided into three pairs, each includes two probes of similar pK(a) in water and different lipophilicity. Solvation has been studied in binary mixtures, BMs, of water, W, with 12 protic organic solvents, S, including mono- and bifunctional alcohols (2-alkoxyethanoles, unsaturated and chlorinated alcohols). Each medium was treated as a mixture of S, W, and a complex solvent, S-W, formed by hydrogen bonding. Values of lambda(max) (of the probe intramolecular charge transfer) were converted into empirical polarity scales, E(T)(probe) in kcal/mol, whose values were correlated with the effective mole fraction of water in the medium, chi w(effective). This correlation furnished three equilibrium constants for the exchange of solvents in the probe solvation shell; phi(W/S) (W substitutes S): phi(S-W/W) (S-W substitutes W), and phi(S-W/S) (S-W substitutes S), respectively. The values of these constants depend on the physicochemical properties of the probe and the medium. We tested, for the first time, the applicability of a new solvation free energy relationship: phi = constant + a alpha(BM) + b beta(BM) + s(pi*(BM) + d delta) + p log P(BM), where a, b, s, and p are regression coefficients alpha(BM), beta(BM), and pi*(BM) are solvatochromic parameters of the BM, delta is a correction term for pi*, and log P is an empirical scale of lipophilicity. Correlations were carried out with two-, three-, and four-medium descriptors. In all cases, three descriptors gave satisfactory correlations; use of four parameters gave only a marginal increase of the goodness of fit. For phi(W/S), the most important descriptor was found to be the lipophilicity of the medium; for phi(S-W/W) and phi(S-W/S), solvent basicity is either statistically relevant or is the most important descriptor. These responses are different from those of E(T)(probe) of many solvatochromic indicators in pure solvents, where the importance of solvent basicity is usually marginal, and can be neglected.

Aspects of Liminality in Eilis Ni Dhuibhne's The Dancers Dancing

In this essay I refer Eilis Ni Dhuibhne’s narrative construction of the main characters and the theme of the novel The Dancers Dancing, in the context of the anthropologist Victor Turner’s concept of liminality. Thus the summer in the Gaeltacht that five teenage girls experience, can be understood as a depiction of the liminal phase in a rite of passage. Ni Dhuibhne’s differently constructed characters enlighten different aspects of liminality and through the céilí dance their experiences are exposed. Furthermore this essay suggests that Julia Kristeva’s notion of the chora, which can be associated to dance, is also relevant when describing the unbounded and unlimited process that radically can reform social structures. I conclude that the liminal space offers an area of many possibilities. It functions as a free zone where the main characters can freely explore their personal issues that trouble them, or the difficulties of their own society.

Poverty as an Abuse of Human Rights in Ghana. : A grass roots perspective on poverty and human rights.

The study aimed at getting a grass root opinion on poverty and why Ghana is still poor after 50 years of independence in spite of her richness in natural resources, second largest producer of cocoa in the word and appreciable stable political environment. The opinions of the ordinary people in the Bia district and their observed living conditions was analysed in line with theoretical basis of the study and previous studies to justify the stance that poverty should be considered as an abuse of human rights. It was concluded based on position of informants and previous data available that though many factors have been raised by previous scholars as the cause of poverty, the actions and inactions of both internal and external power-holders is the main source of poverty in Ghana. It was proposed that for poverty to be reduced in a sustainable way there should be strong civil society groups and active citizens through civic education to hold power-holders accountable. Until the actions and inactions of power-holders which have subjected many Ghanaians into intergenerational poverty are seen as human rights abuse, the rights of many Ghanaians would be constantly abused. This will eventually defeat the promotion of human rights culture in Ghana.

Proceedings from the 2nd seminar on Development of Modular Products : December 13-14, 2004, Campus Framtidsdalen, Dalarna University, Sweden

Companies are focusing on efforts increasing the overall efficiency at the same time as the ability to meet customer needs becomes even more important. There is a need to improve the organisation and the product design at the same time through the visualisation of how a product family design should be performed in order to adapt to customers, company internal issues, and long-term strategy. Therefore, there is a need for qualified personnel in today’s companies with the knowledge of product development and modularity. The graduate course Development of Modular Products at Högskolan Dalarna has the objective to provide such knowledge. As a part of the course, each student will individually perform extensive research within a chosen area with respect to Product Development and Modularity. This proceeding is the result of the students own work and was presented during a two day seminar at Dalarna University. The contents of the papers cover many areas, from the identification of customer needs to cost effective manufacturing, and benefits of modularisation. The reader of this proceeding will not only benefit from many areas within Product Development and Modularity but also from the colour of many cultures. In this proceeding, students from nine countries are represented (Bangladesh, China, Costa Rica, Germany, Holland, India, Luxembourg Nigeria, and Sweden). Enjoy the reading.

Fred: An approach to generating real, correct, reusable programs from proofs

In this paper we describe our system for automatically extracting "correct" programs from proofs using a development of the Curry-Howard process. Although program extraction has been developed by many authors, our system has a number of novel features designed to make it very easy to use and as close as possible to ordinary mathematical terminology and practice. These features include 1. the use of Henkin's technique to reduce higher-order logic to many-sorted (first-order) logic; 2. the free use of new rules for induction subject to certain conditions; 3. the extensive use of previously programmed (total, recursive) functions; 4. the use of templates to make the reasoning much closer to normal mathematical proofs and 5. a conceptual distinction between the computational type theory (for representing programs)and the logical type theory (for reasoning about programs). As an example of our system we give a constructive proof of the well known theorem that every graph of even parity, which is non-trivial in the sense that it does not consist of isolated vertices, has a cycle. Given such a graph as input, the extracted program produces a cycle as promised.

Differential Habituation of Male Betta Splendens to Qualitatively Different Stimuli

Habituation is a learning mechanism that functions to decrease the amount of energy and attention focused on a certain stimuli. Male Siamese Fighting Fish, Betta splendens, are territorial animals that defend their territories using a number of aggressive displays. Male Bettas have previously shown the ability to habituate to the presence of a conspecific male when visually exposed to each other. Due to the costly nature of many of the male Betta’s displays, I hypothesized that male Bettas should differentially habituate to qualitatively different stimuli. I presented each of three groups of male Betta splendens with a different stimulus, each presenting a different level of interactivity. I predicted that the Bettas would be more likely to habituate to a less interactive stimulus than a more interactive one. No significant habituation was observed in any of the groups and no significant differences in latency to display or length of display between all three groups were observed. However, overall data trends suggest that habituation was indeed occurring and that the three different stimuli elicited different levels of display. The limited amount of visual exposure to the stimuli in this experiment might account for why results were insignificant.

The development of a Defense System against Coxsackievirus B5 Infection in Suckling Mice

There are many viruses that are able to infect the alimentary tract of man. Little is known, however, about the mechanism of infection itself or the pathophysiology of the gut during infection. 'The research reported here is concerned with the differences in susceptibility among suckling mice of various ages inoculated by the intraperitoneal and intragastric routes. Since the normal mode of entry of many viruses to the gut is via the oral route, Coxsackievirus B5, a human enterovirus which does attack this way, was utilized. It is a non-tumor producing RNA virus that has been shown to act similarly in the mouse and human. The virus was pooled in HeLa cell cultures and titered by a plaquing assay in the same cell cultures. CD-l mice, 10, 14, 18, and 22 days old , were infected either orally or intraperitoneally with 5.0 x 10^10 (10 day old animals) and 1.0 x10^9 plaque forming units per animal. Dissections were done at 1 and 3 days post infection with samples of the blood, heart, liver, and gut being taken from each animal. Each sample was titered individually and the data presented as an average of six samples. As a result of previous work, it is known that the gut of a newborn mouse isn't able to decrease the concentration of the infecting dose and therefore provides no defense against an enteric infection with Coxsackievirus B5. In contrat, mature mice are able to reduce the amount of viral dissemination across the gut as well as inhibit replication after absorption has occurred. The results of this study indicate that there is a double barrier system developing in suckling mice that is involved with and directly related to the gastrointestinal tract The first part of this defense is the inhibition of penetration of virus across the gut when the primary site of' infection is the intestinal mucosa. This mechanism develops sometime around 20 to 22 days after birth. At about 16-18 days of age, suckling mice that were challenged intragastrically are able to stop active replication and initiate clearance of virus from the systemic circulation. There are many factors that might contribute to the marked decrease in susceptibility with age of suckling mice. Some of these or possibly a combination of these factors might explain the defense mechanisms described above, but to date, the chemistry or mechanical functioning of the gastrointestinal barrier to enteric viral infection is unknown.

O processo de composição narrativa no encontro terapêutico : (des)construindo autorias

Essa pesquisa procurou investigar o processo de composição narrativa pela dupla estagiário-terapeuta/paciente, em uma situação de psicoterapia psicanalítica, a partir do contexto de uma prática supervisionada de estágio em Psicologia Clínica. Participaram da pesquisa duas acadêmicas de Psicologia que realizaram o estágio em um abrigo municipal. O trabalho clínico desenvolvido pelas estagiárias foi acompanhado pela supervisão acadêmica, cuja responsável na época era a pesquisadora. Também participaram dessa pesquisa três meninas de seis, nove e dez anos de idade, acolhidas temporariamente na instituição e em acompanhamento psicoterapêutico pelas estagiárias. Os atendimentos foram realizados uma vez por semana, individualmente, na própria instituição. As estagiárias relataram cada entrevista preliminar realizada com as crianças sob a forma escrita de entrevista dialogada, cujo objetivo é a memorização do desenvolvimento da entrevista. Essa memorização associada às reflexões acerca do estágio produzidas no espaço de supervisão acadêmica formaram as fontes dos dados. Para atingir o objetivo dessa pesquisa, três estudos foram realizados e, em cada um deles, três casos, constituídos por diferentes duplas terapêuticas, foram analisados. Os resultados dos três estudos demonstram, inicialmente, que o discurso elaborado pelas duplas terapêuticas, em cada entrevista preliminar isoladamente, estrutura-se narrativamente porque esse discurso apresenta os dois princípios da narrativa, que são a sucessão e a transformação, como propõe Tzvetan Todorov. A análise conjunta dessas entrevistas denota, entretanto, que as narrativas constituídas nesse processo não podem ser reduzidas a uma lógica de sucessão linear como formula esse autor. A seqüência narrativa é regida pela lógica de causalidade semântica, que é de natureza polifônica, como propõe Paul Ricoeur. As intervenções das estagiárias sob a forma de construções, conforme conceito estabelecido por Freud, mesmo que guiadas pelo princípio da associação livre, são demarcadas, em sua maioria, pela repetição de uma versão já conhecida da história da vida de seu paciente, geralmente àquela que versa sobre o motivo do abrigamento. Assim, essas intervenções, cujo efeito possível seria que o paciente pudesse desconstruir os sentidos dados a priori, reconstruindo novas versões para os acontecimentos de sua vida e, com isso, ocupasse o lugar de autor de sua história, acabam insistindo no trauma. Dessa forma, fica explicitado um dos paradoxos do processo de formação da escuta clínica: o estagiário, ao procurar abrir os sentidos para o seu paciente, construindo junto com ele uma versão possível para a sua história, acaba, muitas vezes, fechando o sentido, construindo uma única versão para os eventos narrados pelo paciente.