Coagulopathy in dengue fever patients

Background:The study was done to find out the prevalence of coagulopathy in Dengue fever patients with thrombocytopenia and its clinical significance.Methods:The patients admitted in medical wards and ICU were included in the study after considering certain inclusion and exclusion criteria. APTT, PT & D-dimer assays were also done in the study population. Coagulopathy was considered when APTT values were ≥41seconds.Results:22.3% of the study population showed evidence of coagulopathy. Bleeding manifestations significantly increased with increasing APTT values. There is also significant association between platelet counts and bleeding manifestations. As platelet count falls there is a tendency for APTT to rise.Conclusion:In addition to thrombocytopenia, coagulopathy also contribute to the presence of bleeding manifestations in dengue fever patients. There is a significant correlation between bleeding manifestations with abnormal APTT values. As platelet count decreases there is tendency for rise in APTT values

LHC Magnet Tests: Operational Techniques and Empowerment for Successful Completion

The LHC magnet tests operation team developed various innovative techniques, particularly since early 2004, to complete the superconductor magnet tests by Feb. 2007. Overall and cryogenic priority handling, rapid on-bench thermal cycling, rule-based goodness evaluation on round-the-clock basis, multiple, mashed web systems are some of these techniques applied with rigour for successful tests completion in time. This paper highlights these operation empowerment tools which had a pivotal role for success. A priority handling method was put in place to enable maximum throughput from twelve test benches, having many different constraints. For the cryogenics infrastructure, it implied judicious allocation of limited resources to the benches. Rapid On-Bench Thermal Cycle was a key strategy to accelerate magnets tests throughput, saving time and simplifying logistics. First level magnet appraisal was developed for 24 hr decision making so as to prepare a magnet further for LHC or keep it on standby. Web based systems (Tests Management and E-Traveller) were other essential ideas to track & coordinate various stages of tests handled by different teams

Therapeutic Targeting of STAT3 (Signal Transducers and Activators of Transcription 3) Pathway Inhibits Experimental Autoimmune Uveitis

Mice with targeted deletion of STAT3 in CD4+ T-cells do not develop experimental autoimmune uveitis (EAU) or experimental autoimmune encephalomyelitis (EAE), in part, because they cannot generate pathogenic Th17 cells. In this study, we have used ORLL-NIH001, a small synthetic compound that inhibits transcriptional activity of STAT3, to ameliorate EAU, an animal model of human posterior uveitis. We show that by attenuating inflammatory properties of uveitogenic lymphocytes, ORLL-NIH001 inhibited the recruitment of inflammatory cells into the retina during EAU and prevented the massive destruction of the neuroretina caused by pro-inflammatory cytokines produced by the autoreactive lymphocytes. Decrease in disease severity observed in ORLL-NIH001-treated mice, correlated with the down-regulation of α4β1 and α4β7 integrin activation and marked reduction of CCR6 and CXCR3 expression, providing a mechanism by which ORLL-NIH001 mitigated EAU. Furthermore, we show that ORLL-NIH001 inhibited the expansion of human Th17 cells, underscoring its potential as a drug for the treatment of human uveitis. Two synthetic molecules that target the Th17 lineage transcription factors, RORγt and RORα, have recently been suggested as potential drugs for inhibiting Th17 development and treating CNS inflammatory diseases. However, inhibiting STAT3 pathways completely blocks Th17 development, as well as, prevents trafficking of inflammatory cells into CNS tissues, making STAT3 a more attractive therapeutic target. Thus, use of ORLL-NIH001 to target the STAT3 transcription factor, thereby antagonizing Th17 expansion and expression of proteins that mediate T cell chemotaxis, provides an attractive new therapeutic approach for treatment of posterior uveitis and other CNS autoimmune diseases mediated by Th17 cells

Application of an in vitro cell monolayer model for evaluating the transport of a peptide and a mineral nutrient.

The overall objectives of this research is two-fold. First, preformulation studies of a novel chemopreventive peptide known as Melanotan-I (MT-I) were performed in order to facilitate the development of suitable formulations of this compound. The studies conducted include (a) the development of a HPLC assay in cell culture transport buffer and human plasma, (b) determination of its partitioning properties and stability kinetics in aqueous solutions, and (c) evaluation of transport properties using an in-vitro cell monolayer model consisting of Caco-2 cells grown as a monolayer on permeable supports. The HPLC method developed here was sensitive, specific and stability-indicating. This assay was successfully applied to the study of MT-I transport across Caco-2 cells. Partitioning of MT-I determined in n-octanol:buffer and isooctane:buffer solvent systems indicated the potential of delivering this peptide via the oral route. MT-I degradation in phosphate buffer exhibited apparent first order kinetics with an estimated shelf life of 40 days at room temperature. It was found to be relatively stable at acidic conditions but had an increased degradation rate at pH > 7.4. Transport of MT-I across Caco-2 cells indicated the degradation of this peptide by the proteases associated with the enterocyte to be the primary obstacle to its oral delivery. MT-I transport was significantly enhanced in the presence of a protease inhibitor (aprotinin). The second objective of this research was to identify enhancers of calcium transport using the in-vitro Caco-2 cell monolayer model. Both medium-chain triglycerides and acylcarnitines were found to enhance the transport of calcium, although to varying degrees. Acylcarnitines were found to be more potent enhancers of calcium transport, but a greater extent of cell damage was observed with their use. In addition, Caco-2 cells were shown to possess L-type calcium channels for the first time, and acylcarnitines were demonstrated to behave like calcium channel agonists similar to that of Bay K 8644 (an established channel agonist). Promotion of both transcellular (membrane perturbation) and paracellular (loosening of tight junctions) pathways of calcium transport were shown to be significant contributors toward the overall enhancement mediated by the acylcarnitines chosen for this study

ORLL-NIH001 inhibited proteins that mediate lymphocyte trafficking into retina.

<p>(A, B) LN cells from mice with EAU were re-stimulated with IRBP in medium containing ORLL-NIH001 or commercially available STAT3 inhibitors. Expression or activation of integrins and chemokine receptors were analyzed by FACS. Plots were gated on CD4⁺ T cells and numbers in quadrants indicate percent of CD4⁺ T cells expressing CD11a, CD29, CD44, CD49d, CXCR3, CCR6, α4β7α4β1 or Ly6c. Data presented are representative of 3 independent experiments.</p

ORLL-NIH001 inhibits expansion of Th17 cells in a dose dependent manner.

<p>(A, B) Naïve CD4⁺ and CD8⁺ T cells were labeled with CFSE, stimulated for 4 days with anti-CD3/anti-CD28 under Th17 polarization condition in medium containing vehicle alone or different doses of ORLL-NIH001 (A). Numbers in the quadrants indicate percent of proliferating or non-proliferating CD4⁺ or CD8+ T cells expressing IL-17. (B) Proliferation of cells in cultures without CFSE was quantified by the Thymidine incorporation assay. Data presented are representative of at least 3 independent experiments.</p

ORLL-NIH001 inhibited expansion of mouse uveitogenic Th17 cells.

<p>(A) Draining LN cells from vehicle-treated (control) or drug-treated mice (Protocol 1) were re-stimulated <i>in vitro</i> with IRBP for 3 days and effects of ORLL-NIH001 was assessed by Thymidine incorporation assay. Freshly isolated PBMC (B, C) was isolated from individual mice 4 days post-immunization. The levels of CD3⁺CD4⁺ T cells were quantified using FACS (B) and the number of cytokine-expressing T cells was assessed by intracellular cytokine assay (C). (D, E) Freshly isolated lymph node cells from vehicle or drug-treated mice were re-stimulated ex vivo with IRBP for 3 days and then analyzed by the intracellular cytokine assay. CFSE was added to some cultures (E). Plots were gated on CD3⁺ T cells and numbers in quadrants indicate percent of CD4⁺ T cells expressing IL-17 and/or IFN-γ. Data presented are representative of at least 3 independent experiments.</p