Differential role of ICAM ligands in determination of human memory T cell differentiation

BACKGROUND: Leukocyte Function Antigen-1 (LFA-1) is a primary adhesion molecule that plays important roles in T cell activation, leukocyte recirculation, and trans-endothelial migration. By applying a multivariate intracellular phospho-proteomic analysis, we demonstrate that LFA-1 differentially activates signaling molecules. RESULTS: Signal intensity was dependent on both ICAM ligand and LFA-1 concentration. In the presence of CD3 and CD28 stimulation, ICAM-2 and ICAM-3 decreased TGFβ1 production more than ICAM-1. In long-term differentiation experiments, stimulation with ICAM-3, CD3, and CD28 generated IFNγ producing CD4+CD45RO+CD62L-CD11a(Bright)CD27- cells that had increased expression of intracellular BCL2, displayed distinct chemokine receptor profiles, and exhibited distinct migratory characteristics. Only CD3/CD28 with ICAM-3 generated CD4+CD45RO+CD62L-CD11a(Bright)CD27- cells that were functionally responsive to chemotaxis and exhibited higher frequencies of cells that signaled to JNK and ERK1/2 upon stimulation with MIP3α. Furthermore, these reports identify that the LFA-1 receptor, when presented with multiple ligands, can result in distinct T cell differentiation states and suggest that the combinatorial integration of ICAM ligand interactions with LFA-1 have functional consequences for T cell biology. CONCLUSION: Thus, the ICAM ligands, differentially modulate LFA-1 signaling in T cells and potentiate the development of memory human T cells in vitro. These findings are of importance in a mechanistic understanding of memory cell differentiation and ex vivo generation of memory cell subsets for therapeutic applications

New treatments in renal cancer: The AhR ligands

Kidney cancer rapidly acquires resistance to antiangiogenic agents, such as sunitinib, developing an aggressive migratory phenotype (facilitated by c-Metsignal transduction). The Aryl hydrocarbon receptor (AhR) has recently been postulated as a molecular target for cancer treatment. Currently, there are two antitumor agent AhR ligands, with activity against renal cancer, that have been tested clinically: aminoflavone (AFP 464, NSC710464) and the benzothiazole (5F 203) prodrug Phortress. Our studies investigated the action of AFP 464, the aminoflavone pro-drug currently used in clinical trials, and 5F 203 on renal cancer cells, specifically examining their effects on cell cycle progression, apoptosis and cell migration. Both compounds caused cell cycle arrest and apoptosis but only 5F 203 potently inhibited the migration of TK-10, Caki-1 and SN12C cells as well as the migration signal transduction cascade, involving c-Met signaling, in TK-10 cells. Current investigations are focused on the development of nano-delivery vehicles, apoferritin-encapsulated benzothiazoles 5F 203 and GW610, for the treatment of renal cancer. These compounds have shown improved antitumor effects against TK-10 cells in vitro at lower concentrations compared with a naked agent.Fil: Itkin, Boris. Gobierno de la Ciudad de Buenos Aires. Hospital General de Agudos "Juan A. Fernández"; ArgentinaFil: Breen, Alastair. University of Nottingham; Estados UnidosFil: Turyanska, Lyudmila. University of Nottingham; Estados UnidosFil: Sandes, Eduardo Omar. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Oncología "Ángel H. Roffo"; ArgentinaFil: Bradshaw, Tracey D.. University of Nottingham; Estados UnidosFil: Loaiza Perez, Andrea Irene. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Oncología "Ángel H. Roffo"; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay; Argentin

Assessment of the toll-like receptor 4 Asp299Gly, Thr399Ile and interleukin-8 -251 polymorphisms in the risk for the development of distal gastric cancer

<p>Abstract</p> <p>Background</p> <p>The intensity of the inflammation induced by <it>Helicobacter pylori </it>colonization is associated with the development of distal gastric cancer (GC). The host response to <it>H</it>. <it>pylori </it>has been related to genetic polymorphisms that influence both innate and adaptive immune responses.</p> <p>Our aim was to investigate whether the presence of the <it>TLR4 Asp299Gly</it>, <it>TLR4 Thr399Ile </it>and <it>IL-8-251 </it>A/T polymorphisms had any influence in the development of distal GC in a Mexican population.</p> <p>Methods</p> <p>We studied 337 patients that were divided in two groups: 78 patients with histologically confirmed distal GC and 259 non-cancer controls. The presence of <it>H. pylori </it>in the control population was defined by positive results of at least two of four diagnostic tests: serology, histology, rapid urease test and culture. Human DNA was purified and genotyped for <it>TLR4 Asp299Gly </it>polymorphism by pyrosequencing, for <it>TLR4 Thr399Ile </it>by PCR-RFLP and for <it>IL8-251 </it>by the amplification refractory mutation system (ARMS)-PCR.</p> <p>Results</p> <p>The non-cancer control group was found to be in Hardy-Weinberg equilibrium at the polymorphic loci studied (chi-square _H-W= 0.58 for <it>IL8-251</it>, 0.42 for <it>TLR4 Asp299Gly </it>and 0.17 for <it>TLR4 Thr399Ile</it>). The frequencies of mutated alleles (homozygous plus heterozygous) were compared between cases and controls. We found no significant difference for <it>TLR4- Asp299Gly </it>[the 7.7% of distal GC patients and 7.7 % non-cancer controls (p = 0.82)] and for <it>TLR4 Thr399Ile </it>[the 1.3% of GC patients and the 5% of the control population (p = 0.2)]. In contrast, for <it>IL-8-251 </it>A/T, 80.77% of the GC patients and 66.4% in the control group age and gender matched had at least one copy of mutated allele (OR = 2.12, 95% CI = 1.1–4.2) (p = 0.023).</p> <p>Conclusion</p> <p>This study showed that the <it>IL8-251*A </it>allele could be related to the development of distal gastric cancer in this Mexican population.</p

Accelerated directed evolution of dye-decolorizing peroxidase using a bacterial extracellular protein secretion system (BENNY)

Background Dye-decolorizing peroxidases (DyPs) are haem-containing peroxidases that show great promises in industrial biocatalysis and lignocellulosic degradation. Through the use of Escherichia coli osmotically-inducible protein Y (OsmY) as a bacterial extracellular protein secretion system (BENNY), we successfully developed a streamlined directed evolution workflow to accelerate the protein engineering of DyP4 from Pleurotus ostreatus strain PC15. Result After 3 rounds of random mutagenesis with error-prone polymerase chain reaction (epPCR) and 1 round of saturation mutagenesis, we obtained 4D4 variant (I56V, K109R, N227S and N312S) that displays multiple desirable phenotypes, including higher protein yield and secretion, higher specific activity (2.7-fold improvement in kcat/Km) and higher H2O2 tolerance (sevenfold improvement based on IC50). Conclusion To our best knowledge, this is the first report of applying OsmY to simplify the directed evolution workflow and to direct the extracellular secretion of a haem protein such as DyP4

Determinación y Estandarización De Las Mejores Condiciones De Proceso De Lavado Cip (Cleaning In Place): En La Zona De Evaporación De La Planta Pulverizadora De Leche Colanta Planeta Rica

La industria lechera utiliza desde hace muchos años el CIP (cleaning in place) como método de limpieza. En la planta Pulverizadora Colanta Planeta Rica no existía un procedimiento de CIP que garantizara la eficiencia y condiciones constantes durante el proceso. Mediante modificaciones de infraestructura en los equipos, recomendaciones de los proveedores, revisión bibliográfica y ensayos realizados en la planta se logró determinar y estandarizar las condiciones para desarrollar el proceso CIP como; concentraciones y temperaturas de las soluciones de lavado, caudales y tiempos de proceso. Se logró que las soluciones de lavado recircularan a las concentraciones deseadas (soda caustica de 3 – 4% y ácido nítrico de 2 – 3%). Se consiguió mantener la temperatura de la soda en un rango de 70 a 80o C durante su recirculación en los efectos del evaporador. Se alcanzó el caudal máximo posible para realizar el CIP en el pasterizador el cual garantizara el arrastre total de partículas adheridas y se determinó el caudal más adecuado para el lavado del evaporador que garantizara una limpieza eficiente y evitara la inundación del equipo. Se logró la reducción del CIP de 6 a 4,5 horas representando una ganancia de $748 ́459.008 anuales, ya que el tiempo ahorrado es ahora invertido en la producción de leche en polvo, lo cual estuvo mayoritariamente influenciado por la reducción del tiempo de recirculación de soda en el pasterizador, el cual no se había bajado de 3 horas pese a la sospecha que pudiera influir negativamente en el equipo o en el proceso productivo

Caracterización del Desbalance en Redes de Distribución Eléctricas Argentinas, a través del Factor de Desbalance Contemplando la Reglamentación Vigente

El objeto del presente trabajo es mostrar cuales son los márgenes de desbalance que aparecen y/o podrían aparecer en el sistema de distribución de la Argentina, sin violar la reglamentación nacional vigente al respecto, para compararlos con los recomendados por la normativa internacional vinculada con el tema. En tal sentido se determina el máximo valor posible que podría alcanzar la relación porcentual entre la componente de secuencia negativa y positiva (relación empleada por IEC e IEEE para evaluar el desbalance), utilizando como herramienta un algoritmo genético diseñado específicamente para cumplir con lo propuesto. Además se muestran una serie de mediciones de desbalance, obtenidas en distintos puntos del sistema de distribución eléctrica de baja tensión

The control and maintenance of desired flow patterns in bends of different orientations

Multiphase flows are common in industrial settings and bends in pipe lines cannot be avoided due to space limitations. Gas-liquid two phase flows could form material discontinuities that could have adverse effect on productivity and the pipe network due to sudden variations resulting due to the rapid momentum flux variations at fittings such as bends. Research into gas-liquid flow and bends can be motivated by the effect of the bend on the flow downstream of it which could alter the flow pattern occurring and the performance of downstream equipment. Alternatively, the interest might come from what occurs in the bend itself, there could be dryout of the film on the walls and consequent damage to the heat transfer equipment. Here we present measurements made with a number of accurate and fast responding sensors on three cases, two on the effect of the bend and one considering effects in the bend. The results show that the flow transformations occur in two phase flows depending on the orientation of the bend and the change could be captured using fast sweeping measurement techniques. We present the evidence of effectiveness of several types of measurement techniques that could fit into various combinations of phases. The results, point to how to achieve certain flow patterns. Also recommendations are provided regarding the position of any sensor installed to determine flow pattern

Discusión de la teoría económica y su aplicación en modelos multidisciplinarios energéticos : Parte 1: Introducción a los métodos de decisión difusos

This paper deals with introductory aspects of fuzzy decision theory: law of supply and demand, market and its limitations (externalities) with the solutions proposed in theory and practice to solve them (tolls). Within these limitations (still under discussion), the concept of uncertainty is introduced, an aspect that hinders an optimal decision making of the participating individuals based on current economic theories that are based on the full certainty of the economic context. Therefore, decision theory and its extension to fuzzy decision theory, which is applicable in contexts of uncertainty, are introduced. The Analytic Hierarchy Process (AHP) and its link with fuzzy decision models by means of exponential weights are presented. Therefore, the purpose of this article is to define a methodology for optimal evaluation, valuation and regulation, in contexts of fundamental uncertainty, determining the optimal reward or punishment, based on non-monetizable indexes (directly translatable to an equivalent economic cost). From there, an experimental index is defined, which is used in the present line of research, the Intrinsic Cost (IC).En el presente trabajo se abordan aspectos introductorios a la teoría de decisión difusa: ley de oferta y demanda, mercado y sus limitaciones (externalidades) con las soluciones propuestas en la teoría y práctica para solucionarlas (peaje). Dentro de estas limitaciones (aún en discusión), se introduce el concepto de incertidumbre, aspecto que dificulta una toma de decisión óptima de los individuos participantes en base a las teorías económicas actuales que se basan en la plena certidumbre del contexto económico. Por lo tanto, se introduce la teoría de decisión y su extensión hacia la teoría de la decisión difusa, que es aplicable en contextos de incertidumbre. Se presenta el Análisis de Procesos Jerárquicos – Analytic Hierarchy Process (AHP) y su vínculo con los modelos de decisión difusos mediante los ponderadores exponenciales. Por lo tanto, el propósito del presente artículo es definir una metodología de evaluación, valoración y regulación óptima, en contextos de incertidumbre del tipo fundamental, determinando el premio o castigo óptimo, en base a índices no monetizables (traducibles a un costo económico equivalente) de forma directa. A partir de allí se define un índice experimental, el cual es utilizado en la presente línea de investigación, el Costo Intrínseco – Intrinsic Cost (IC). ARK CAICYT: http://id.caicyt.gov.ar/ark:/s18539777/r6x80gjkr

Rehabilitation of torture survivors in five countries: common themes and challenges

<p>Abstract</p> <p>Background</p> <p>Torture continues to be a global problem and there is a need for prevention and rehabilitation efforts. There is little available data on torture survivors from studies designed and conducted by health professionals in low income countries. This study is a collaboration between five centres from Gaza, Egypt, Mexico, Honduras and South Africa who provide health, social and legal services to torture survivors, advocate for the prevention of torture and are part of the network of the International Rehabilitation Council for Torture Victims (IRCT).</p> <p>Methods</p> <p>Socio-demographic, clinical and torture exposure data was collected on the torture survivors attending the five centres at presentation and then at three and six month follow-up periods. This sample of torture survivors is presented using a range of descriptive statistics. Change over time is demonstrated with repeated measures analysis of variance.</p> <p>Results</p> <p>Of the 306 torture survivors, 23% were asylum seekers or refugees, 24% were socially isolated, 11% in prison. A high level of traumatic events was experienced. 64% had suffered head injury whilst tortured and 24% had ongoing torture injury problems. There was high prevalence of symptoms of anxiety, depression, post traumatic stress as well as medically unexplained somatic symptoms. The analysis demonstrates a modest drop in symptoms over the six months of the study.</p> <p>Conclusions</p> <p>Data showed that the torture survivors seen in these five centres had high levels of exposure to torture events and high rates of clinical symptoms. In order to provide effective services to torture survivors, health professionals at torture rehabilitation centres in low income countries need to be supported to collect relevant data to document the needs of torture survivors and to evaluate the centres' interventions.</p