Induced Pluripotent Stem Cells Technology For Investigating Familial Dilated Cardiomyopathy Due To Lamin A/C Mutations

Mutations of LMNA gene, encoding the proteins Lamin A and C, are common causes of familial dilated cardiomyopathy (DCM), typically manifesting in association with cardiac conduction defects. LaminA/C regulate various nuclear functions, including gene transcription and chromatin organization. Most of human studies on LaminA/C so far were conducted on fibroblasts, while those focusing on cardiomyocytes (CMs) are scarce. Thus, we generated a human cardiac model of LMNA-dependent cardiomyopathy (LMNA-CMP) by differentiating CMs from induced pluripotent stem cells (iPSCs) carrying the K219T and R190W LMNA mutations (LMNA-CMs). In vitro, these cells recapitulate morphological features of DCM. RNA-sequencing experiments revealed profound differences between control and LMNA-CMs transcriptomes. Remarkably, the main cardiac sodium channel gene (SCN5A), was one of the most downregulated gene. Accordingly, electrophysiological studies in LMNA-CMs showed a significant reduction of the maximal upstroke velocity and the related properties of the action potential, accompanied by a reduction of the peak sodium currents and diminished conduction velocity. Functional experiments also revealed diminished contractile force in LMNA mutated cells. Biochemical studies confirmed the reduction of SCN5A gene, and its encoded protein Nav1.5, and showed an increased binding of LaminA/C to its promoter. A higher deposition of H3K27me3 marker, together with an increased binding of its catalysing complex PRC2, was also found at that genomic locus. Furthermore, 3D-FISH studies indicated a preferential localization of SCN5A gene at the nuclear periphery. Similar findings were obtained in relation to genes of the contractile pathways. Altogether, these data support a mechanism by which LaminA/C influence cardiac function by directly acting on transcriptional regulation of genes that control CMs functionality. Studies conducted on CMs from CRISPR/Cas9 gene-edited iPSCs proved the causality between the LMNA mutation and the observed phenotypes. Finally, experiments for evaluating the efficacy of Remodelin molecule also demonstrated that the generated models are suitable for drug testing

Relação entre a unidade de biofeedback de pressão, deslocamento do centro de pressão e atividade eletromiográfica durante o active straight leg raising em indivíduos com e sem dor lombo-pélvica.

Introdução: A unidade de biofeedback de pressão, em indivíduos com dor lombo-pélvica, é utilizada durante exercícios de estabilização segmentar, no entanto ainda carece de evidência científica. Objetivo: Determinar a relação entre a unidade de biofeedback de pressão (UBFP), o deslocamento do centro de pressão no sentido médio-lateral (COPml) e a atividade eletromiográfica abdominal durante o active straight leg raising (ASLR) em indivíduos com e sem dor lombo-pélvica, bem como identificar diferenças entre os grupos. Metodologia: Estudo transversal analítico em 18 estudantes universitários voluntários com dor lombo-pélvica crónica inespecífica (GCD) e em 20 sem dor (GSD). Durante o ASLR (desafio postural dinâmico) foram avaliadas as variações máxima e média da pressão (recorrendo à UBFP) e do deslocamento do COPml (através da plataforma de forças), bem como a atividade muscular abdominal, bilateralmente, com recurso à eletromiografia de superfície. Estatisticamente recorreu-se à correlação de Spearman e ao teste Mann-Whitney U, ambos com um nível de significância de 0,05. Resultados: No GCD, ao contrário do GSD, não foi verificada uma relação entre a UBFP e a atividade do transverso abdominal/obliquo interno (TrA/OI) contra-lateral. Correlações moderadas, mas com sentidos opostos, foram evidenciadas em ambos os grupos, entre o deslocamento do COPml e a atividade do TrA/OI contra-lateral. Em ambos os grupos, a UBFP demonstrou estar fortemente correlacionada com o COPml. Não foram observadas diferenças significativas entre os grupos nas variáveis avaliadas. Conclusão: A UBFP, no GCD, não se apresentou relacionada com a atividade do TrA/OI. Contudo, demonstrou uma relação com o deslocamento do COPml, em ambos os grupos, sendo portanto um indicador de estabilidade do tronco e assim, uma ferramenta útil em ambiente clínico. No GCD observou-se que uma maior atividade muscular TrA/OI pressupõe maior deslocamento do COPml, sendo uma relação contrária à verificada no GSD, podendo ser um indicador da perda da sua ação tónica.Background: Pressure biofeedback unit, in subjects with low back pain, is used during segmental stabilization exercises, however scientific evidence is still lacking. Objective: Determine the relationship between pressure biofeedback unit (UBFP), center of pressure medial-lateral displacement (COPml) and electromyographic activity of the abdominal muscles during the active straight leg raising (ASLR) in individuals with and without low back pain, as well as identify differences between groups. Methods: Analytical cross-sectional study in 18 college students volunteers with chronic nonspecific low back pain (GCD) and 20 without pain (GSD). During the ASLR (dynamic postural challenge) were evaluated pressure (using the UBFP) and COPml displacement's mean and maximal variation, as well as bilateral abdominal muscles activity using surface electromyography. Statistically it were used Spearman correlations and Mann-Whitney U tests, both with a significance level of 0,05. Results: Unlike GSD, in the GCD it was not found a relationship between UBFP and contra-lateral transversus abdominis/internal oblique (TrA/OI) muscle activity. Moderate correlations, but with opposite directions, were found in both groups between COPml displacement and contra-lateral TrA/OI activity. In both groups, UBFP showed to be strongly correlated with COPml. There were no significant differences between groups in the evaluated variables. Conclusion: The UBFP, in GCD, did not appear to be related with TrA/OI activity. However, it was found a relationship with COPml displacement in both groups, thus being an indicator of trunk stability and so can be useful in the clinical environment. In GCD it was observed that a greater TrA/OI muscle activity implies a greater COPml displacement, being contrary to the relationship found in GSD, and could be an indicator of their loss of tonic action

Structural Analysis Of Bulky Thiolate-Protected Gold Nanomolecules

Routine preparation of thiolate-protected gold nanomolecules involves, most commonly, straight-chain organic thiolate ligands for which the sulfhydryl `head\u27 group is bound to a primary carbon atom. The primary connectivity of these sulfur atoms, such as for the case of n-hexanethiol and phenylethanethiol, allows for even packing, and the relative ease of formation of gold-thiolate oligomers about the surface monolayer in the formation of Au-SR bonds. Moreover, these straight-chain ligands routinely support the formation of an icosahedron-based series of core-size gold nanomolecules. Within this series of nanoparticles, Au25(SR)18 and Au38(SR)24 are among the most commonly studied core-sizes; Au25(SR)18 is situated upon an Au13 icosahedron, and Au38(SR)24 upon two face-fused icosahedra which share three gold atoms to give an Au23 core structure. The investigation of the experimental results presented by the student in this thesis outlines the alteration of these icosahedron-based core geometries. Specifically, by varying the steric hindrance of the –R group for which the sulfur head group is bound, a heightened gold:thiol ratio is achieved. With the application of bulky ligands, most notably tert-butanethiol and 1-adamantanethiol, the underlying geometry is altered such that face-centered cuboidal (fcc), octahedron-based structures are favored. These structures are defined by atomic compositions, predicted by mass spectrometry and confirmed by X-ray Diffraction methods, for which a fewer number of sulfur ligands are needed to stabilize the overall structure. This is best illustrated in the case of the Au25(S-C2H4)18 and the Au30(S-t-C4H9)18 nanomolecules; because both have the same number of stabilizing ligands, it is clear that the addition of five gold atoms to the composition is accompanied by a structural alteration. In this work, the research and discussion focuses on the structural alteration associated with the 3-dimensional atomic arrangement in employment of bulky ligands, with experimentally-dervived data acquired from mass spectrometry, UV-visible spectroscopy, and single-crystal X-ray Diffraction

THE SMART BOOKSHELF

The smart bookshelf serves as a test-bed to study environments that are intelligently augmented by projector-camera devices. The system utilizes a camera pair and a projector coupled with an RFID reader to monitor and maintain the state of a real world library shelf. Using a simple calibration scheme, the homography induced by the world plane in which book spines approximately lie is estimated. As books are added to the shelf, a foreground detection algorithm which takes into account the projected information yields new pixels in each view that are then verified using a planar parallax constraint across both cameras to yield the precise location of the book spine. The system allows users to query for the presence of a books through a user interface, highlighting the spines of present book using the known locations obtained through foreground detection and transforming image pixels to their corresponding points in the projectors frame via a derived homography. The system also can display the state of the bookshelf at any time in the past. Utilizing RFID tags increases robustness and usefulness of the application. Tags encode information about a book such as the title, author, etc, that can be used to query the system. It is used in conjunction with the visual system to infer the state of the shelf. This work provides a novel foreground detection algorithm that works across views, using loose geometric constraints instead pixel color similarity to robustly isolate foreground pixels. The system also takes into account projected information which if not handled would be detrimental to the system. The intent of this work was to study the feasibility of an augmented reality system and use this application as a testbed to study the issues of building such a system

The Various Methods Circuit Courts use to Define Initial Transferee in Fraudulent Transfers

(Excerpt) Transfers of a debtor\u27s interest or obligation in property to a third party, made to prevent creditors from reaching assets in a bankruptcy case, are known as fraudulent transfers. Under current law, there are two types of fraudulent transfers: actual fraud and constructive fraud. Actual fraud requires findings of a debtor\u27s intent to hinder, delay, or defraud any entity to which the debtor was or became, on or after the date that such transfer was made or such obligation was incurred, indebted.\u22 Constructive fraud does not require a finding of intent and occurs when a debtor receives less than a reasonably equivalent value in exchange for such transfer or obligation and if the debtor was insolvent on the date that such transfer was made or such obligation was incurred, or became insolvent as a result of such transfer or obligation. Under Section 548 of Title 11 of the United States Code (the Bankruptcy Code ), trustees have the power to avoid fraudulent transfers made within two years of a bankruptcy proceeding

“Globally Local” CSR in MNC’s : how MNC’s balance the pressures for global integration and local responsiveness in their CSR strategy?

This research aims to advance the discussion of how multinational companies balance global integration with local responsiveness in their corporate social responsibility (CSR) strategy. In particular, it studies the mechanisms between home and host countries in the development and implementation processes of a transnational CSR strategy and the types of monitoring mechanisms used. Structured by a qualitative study with a case study on a hospitality group, we find that the transnational approach is the most suitable indeed to guarantee a good balance between global standardization and local reactiveness. We advocate so by proposing some key mechanisms of a transnational CSR strategy that allow for this balance, and we discuss the main tensions and solutions that arise from this strategy. Finally, we suggest that the transnational approach to CSR should not be untied to local and global approaches and that the integration of the three is what makes the transnational approach the most suitable for MNC’s that want to balance global integration with local responsiveness.O objetivo deste estudo é de desenvolver e fazer progredir o debate sobre como empresas multinacionais podem equilibrar integração global e capacidade de reação e resposta a nivel local na sua estratégia de responsabilidade social. Em particular, pretende estudar os diferentes mecanismos utilizados entre a sede e subisidiárias nos processos de desenvolvimento e implementação de uma estratégia de responsabilidade social transnacional e os tipos de mecanismos de monitoração utilizados. Estruturado por um estudo de caso sobre um grupo hoteleiro, descobrimos que a abordagem transnacional é a mais adequada para garantir um bom equilíbrio entre a integração global e a reatividade local. Defendemos este argumento propondo alguns mecanismos-chave de uma estratégia transnacional de responsabilidade social que permitem garantir esse equilíbrio e discutimos as principais tensões e soluções que surgem neste tipo de estratégia. Finalmente, sugerimos que a estratégia de responsabilidade transnacional não deve ser desvinculada das abordagens locais e globais e que a integração das três torna a abordagem transnacional mais adequada para as multinacionais que desejam equilibrar a integração global com a capacidade de resposta local

An Olfactory Receptor Pseudogene whose Function emerged in Humans

Human olfactory receptor, hOR17-210, is identified as a pseudogene in the human genome. Experimental data has shown however, that the gene product of cloned hOR17-210 cDNA was able to bind an odorant-binding protein and is narrowly tuned for excitation by cyclic ketones. Supported by experimental results, we used the bioinformatics methods of sequence analysis, computational protein modeling and docking, to show that functionality in this receptor is retained due to sequence-structure features not previously observed in mammalian ORs. This receptor does not possess the first two transmembrane helical domains (of seven typically seen in GPCRs). It however, possesses an additional TM that has not been observed in other human olfactory receptors. By incorporating these novel structural features, we created two putative models for this receptor. We also docked odor ligands that were experimentally shown to bind hOR17-210 model. We show how and why structural modifications of OR17-210 do not hinder this receptor's functionality. Our studies reveal that novel gene rearrangement that result in sequence and structural diversity in has a bearing on OR and GPCR function and evolution

Beyond Modeling: All-Atom Olfactory Receptor Model Simulations

Olfactory receptors (ORs) are a type of GTP-binding protein-coupled receptor (GPCR). These receptors are responsible for mediating the sense of smell through their interaction with odor ligands. OR-odorant interactions marks the first step in the process that leads to olfaction. Computational studies on model OR structures can generate focused and novel hypotheses for further bench investigation by providing a view of these interactions at the molecular level beyond inferences that are drawn merely from static docking. Here we have shown the specific advantages of simulating the dynamic environment associated with OR-odorant interactions. We present a rigorous protocol which ranges from the creation of a computationally derived model of an olfactory receptor to simulating the interactions between an OR and an odorant molecule. Given the ubiquitous occurrence of GPCRs in the membranes of cells, we anticipate that our OR-developed methodology will serve as a model for the computational structural biology of all GPCRs