Design and Development of a Hand Exoskeleton Robot for Active and Passive Rehabilitation

The present work, which describes the mechatronic design and development of a novel rehabilitation robotic exoskeleton hand, aims to present a solution for neuromusculoskeletal rehabilitation. It presents a full range of motion for all hand phalanges and was specifically designed to carry out position and force-position control for passive and active rehabilitation routines. System integration and preliminary clinical tests are also presented

Differential Role of Human Choline Kinase α and β Enzymes in Lipid Metabolism: Implications in Cancer Onset and Treatment

11 pages, 6 figures, 1 table.Background The Kennedy pathway generates phosphocoline and phosphoethanolamine through its two branches. Choline Kinase (ChoK) is the first enzyme of the Kennedy branch of synthesis of 1phosphocholine, the major component of the plasma membrane. ChoK family of proteins is composed by ChoKα and ChoKβ isoforms, the first one with two different variants of splicing. Recently ChoKα has been implicated in the carcinogenic process, since it is over-expressed in a variety of human cancers. However, no evidence for a role of ChoKβ in carcinogenesis has been reported. Methodology/Principal Findings Here we compare the in vitro and in vivo properties of ChoKα1 and ChoKβ in lipid metabolism, and their potential role in carcinogenesis. Both ChoKα1 and ChoKβ showed choline and ethanolamine kinase activities when assayed in cell extracts, though with different affinity for their substrates. However, they behave differentially when overexpressed in whole cells. Whereas ChoKβ display an ethanolamine kinase role, ChoKα1 present a dual choline/ethanolamine kinase role, suggesting the involvement of each ChoK isoform in distinct biochemical pathways under in vivo conditions. In addition, while overexpression of ChoKα1 is oncogenic when overexpressed in HEK293T or MDCK cells, ChoKβ overexpression is not sufficient to induce in vitro cell transformation nor in vivo tumor growth. Furthermore, a significant upregulation of ChoKα1 mRNA levels in a panel of breast and lung cancer cell lines was found, but no changes in ChoKβ mRNA levels were observed. Finally, MN58b, a previously described potent inhibitor of ChoK with in vivo antitumoral activity, shows more than 20-fold higher efficiency towards ChoKα1 than ChoKβ. Conclusion/Significance This study represents the first evidence of the distinct metabolic role of ChoKα and ChoKβ isoforms, suggesting different physiological roles and implications in human carcinogenesis. These findings constitute a step forward in the design of an antitumoral strategy based on ChoK inhibition.This work has been supported by grants to JCL from Comunidad de Madrid (GR-SAL-0821-2004), Ministerio de Ciencia e Innovación (SAF2008-03750, RD06/0020/0016), Fundación Mutua Madrileña, and by a grant to ARM from Fundación Mutua Madrileña.Peer reviewe

Severe Asthma Standard-of-Care Background Medication Reduction With Benralizumab: ANDHI in Practice Substudy

Background: The phase IIIb, randomized, parallel-group, placebo-controlled ANDHI double-blind (DB) study extended understanding of the efficacy of benralizumab for patients with severe eosinophilic asthma. Patients from ANDHI DB could join the 56-week ANDHI in Practice (IP) single-arm, open-label extension substudy. Objective: Assess potential for standard-of-care background medication reductions while maintaining asthma control with benralizumab. Methods: Following ANDHI DB completion, eligible adults were enrolled in ANDHI IP. After an 8-week run-in with benralizumab, there were 5 visits to potentially reduce background asthma medications for patients achieving and maintaining protocol-defined asthma control with benralizumab. Main outcome measures for non-oral corticosteroid (OCS)-dependent patients were the proportions with at least 1 background medication reduction (ie, lower inhaled corticosteroid dose, background medication discontinuation) and the number of adapted Global Initiative for Asthma (GINA) step reductions at end of treatment (EOT). Main outcomes for OCS-dependent patients were reductions in daily OCS dosage and proportion achieving OCS dosage of 5 mg or lower at EOT. Results: For non-OCS-dependent patients, 53.3% (n = 208 of 390) achieved at least 1 background medication reduction, increasing to 72.6% (n = 130 of 179) for patients who maintained protocol-defined asthma control at EOT. A total of 41.9% (n = 163 of 389) achieved at least 1 adapted GINA step reduction, increasing to 61.8% (n = 110 of 178) for patients with protocol-defined EOT asthma control. At ANDHI IP baseline, OCS dosages were 5 mg or lower for 40.4% (n = 40 of 99) of OCS-dependent patients. Of OCS-dependent patients, 50.5% (n = 50 of 99) eliminated OCS and 74.7% (n = 74 of 99) achieved dosages of 5 mg or lower at EOT. Conclusions: These findings demonstrate benralizumab's ability to improve asthma control, thereby allowing background medication reduction

XVI International Congress of Control Electronics and Telecommunications: "Techno-scientific considerations for a post-pandemic world intensive in knowledge, innovation and sustainable local development"

Este título, sugestivo por los impactos durante la situación de la Covid 19 en el mundo, y que en Colombia lastimosamente han sido muy críticos, permiten asumir la obligada superación de tensiones sociales, políticas, y económicas; pero sobre todo científicas y tecnológicas. Inicialmente, esto supone la existencia de una capacidad de la sociedad colombiana por recuperar su estado inicial después de que haya cesado la perturbación a la que fue sometida por la catastrófica pandemia, y superar ese anterior estado de cosas ya que se encontraban -y aún se encuentran- muchos problemas locales mal resueltos, medianamente resueltos, y muchos sin resolver: es decir, habrá que rediseñar y fortalecer una probada resiliencia social existente - producto del prolongado conflicto social colombiano superado parcialmente por un proceso de paz exitoso - desde la tecnociencia local; como lo indicaba Markus Brunnermeier - economista alemán y catedrático de economía de la Universidad de Princeton- en su libro The Resilient Society…La cuestión no es preveerlo todo sino poder reaccionar…aprender a recuperarse rápido.This title, suggestive of the impacts during the Covid 19 situation in the world, and which have unfortunately been very critical in Colombia, allows us to assume the obligatory overcoming of social, political, and economic tensions; but above all scientific and technological. Initially, this supposes the existence of a capacity of Colombian society to recover its initial state after the disturbance to which it was subjected by the catastrophic pandemic has ceased, and to overcome that previous state of affairs since it was found -and still is find - many local problems poorly resolved, moderately resolved, and many unresolved: that is, an existing social resilience test will have to be redesigned and strengthened - product of the prolonged Colombian social conflict partially overcome by a successful peace process - from local technoscience; As Markus Brunnermeier - German economist and professor of economics at Princeton University - indicates in his book The Resilient Society...The question is not to foresee everything but to be able to react...learn to recover quickly.Bogot

Comparative expression of ChoKα1 and ChoKβ1 mRNA in a panel of cancer cell lines.

<p>Q-PCR was performed to determine the level of expression of mRNA in non-tumorogenic mammary cell lines (HMEC, MCF10A), breast cancer cell lines (MDA-MB435, MDA-MB468, T47D, MCF7), non-tumorogenic lung cells (BEC) and lung cancer cell lines (H510, H82). The data were normalized with the endogenous 18S ribosomal RNA. For the comparison between tumoral and non-tumoral cell lines, the 2^−ΔΔCt method was applied and log₁₀ RQ is represented. Note that the data are referred to the Human Mammary Epithelial Cells (HMEC) mRNA levels in breast cell lines and no significant difference in the level of both ChoK isoforms mRNA was found with the normal MCF10A cell line. The reference for lung cancer cells was the primary Bronchial Epithelial Cells (BEC).</p

Overexpression of ChoKβ1 is not sufficient to induce tumor growth in athymic nude mice.

<p>Xenografts were established by s.c. injection of transfected HEK293T or MDCK cells in athymic nu/nu nude mice. <b>A)</b> and <b>B)</b> Western Blot analysis of ectopic expression of choline kinase isoforms in transfected HEK293T or MDCK cells, respectively, before mice inoculation. <b>C)</b> and <b>D)</b> Analysis of choline kinase activity in ChoKα1 or β1 transfected HEK293T or MDCK cells-free extracts before mice inoculation. <b>E)</b> and <b>F)</b> Volume of tumors generated by subcutaneous injection of 10⁶ transfected cells. Tumoral volume was calculated according to the formula: <i>Vol = [D * d²]/2</i>, where D and d are major and minor tumor diameters respectively. The data from HEK293T represents mean values±SEM from two independent experiments (n₁ = 12; n₂ = 16), the MDCK experiment correspond to an equivalent experiment with n = 12.</p

Anchorage independent cell growth of ChoKα1- and β1-overexpressing cells.

<p><b>A)</b> and <b>B)</b><i>In vitro</i> ChoK activity of cell-free extracts from transfected cells at the moment of plating, determined as conversion of ¹⁴C-labeled choline to PCho. <b>C)</b> Photographs of a representative experiment of the soft agar assay. A total of 10⁵ cells were plated per 60-mm dish, and the number of colonies quantified after 5-8 weeks of incubation. <b>D)</b> and <b>E)</b> Computer based automatic quantification of the number of colonies, mean values±SEM is represented. The assay was performed 3 independent times with triplicate samples obtaining similar results. Statistical significance (p≤0.05) is marked by an asterisk.</p

Differential activation of choline kinase α1 and β1 isoforms by Ras and Rho GTPases.

<p>Choline kinase isoforms were expressed alone or in combination with the indicated Ras and Rho GTPases and the <i>in vitro</i> ChoK activity determined. <b>A)</b> Analysis of ectopic expression by Western Blot in HEK293T transfected cell extracts of ChoKα1 (52 KDa), ChoKβ1(45 KDa), RhoA-QL(22 KDa), Cdc42-QL(25 KDa) and H-rasV12 (23 KDa). Empty vectors were used as controls for the endogenous levels, and GAPDH as loading control. <b>B)</b> and <b>C)</b><i>In vitro</i> choline kinase activity of ChoKα1 or ChoKβ1 in the presence of enhanced expression of constitutive active forms of RhoA, Cdc42 or H-Ras. <b>D)</b> and <b>E)</b><i>In vitro</i> ethanolamine kinase activity of ChoKα or ChoKβ in the presence of each indicated constitutive active form of GTPase. The results are represented as fold induction of conversion to the corresponding phosphorylated metabolite determined as total cpm/µg of whole cellular extract, and normalized to the empty vector transfected cells as control. Data shown represent the mean values±SEM of 3 independent experiments, each one performed with duplicate samples. Statistical significance (p≤0.05) is marked by an asterisk comparing to the activity achieved when ChoKα1 or ChoKβ1, where appropriate, are transfected alone.</p

Michaelis constant (Km) of ChoKα and β isoforms for choline and ethanolamine.

1<p>Data are represented in milliMolar.</p>2<p>Referenced to the lowest Km.</p><p>Km of the different isoforms of ChoK for each substrate is indicated in each case. The results were obtained from four independent experiments using the logarithmic Michaelis-Menten formula as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007819#s4" target="_blank">Material and Methods</a>.</p

Characterization of choline and ethanolamine kinase activity of ChoKα1 and Chokβ1 in HEK293T cells.

<p>HEK293T cells were transfected with eukaryotic expression vectors of human ChoKα1 and ChoKβ1 gene. pCDNA3b empty vector was used as control. <b>A)</b> Overexpression of ChoKα1 and ChoKβ1 in HEK293T cells detected by Western Blot. GAPDH detection was used as control of expression level. <b>B, C)</b> In vitro ChoK (B) and EtnK (C) activity of choline kinase α1 and β1 isoforms in cell-free extracts of HEK293T transfected cells. Percentage of conversion of ¹⁴C-choline or ¹⁴C-ethanolamine to the phosphorylated product is represented. The experiment was performed in duplicate samples, repeated 4 times, and mean±SEM values from all experiments estimated.</p