In vitro flow evaluation of recombinant tissue factor pathway inhibitor immobilized on collagen impregnated Dacron

Recombinant tissue factor pathway inhibitor (rTFPI) was immobilized on collagen impregnated (CI) knitted Dacron surfaces and its resistance to fibrin deposition evaluated following exposure to nonanticoagulated whole blood. Recombinant TFPI readily adsorbed to the CI Dacron surface and maintained its inhibitory activity. Under static conditions, rTFPI treated CI Dacron showed little fibrin deposition when compared with untreated surfaces. Treated samples exposed to flowing native blood at wall shear rates of 100 or 200 sec(-1) also demonstrated reduced fibrin deposition (up to 56%) compared with untreated samples. To assess the relative roles of the contact and tissue factor pathways in fibrin formation on artificial grafts, flow studies were performed with whole blood containing corn trypsin inhibitor, a potent inhibitor of FXIIa and contact activation. Corn trypsin inhibitor reduced fibrin deposition on untreated CI Dacron by 40%. Immobilized rTFPI alone, or corn trypsin inhibitor in combination with immobilized rTFPI, reduced fibrin deposition by 58% and 61%, respectively. These data suggest that immobilized rTFPI slows fibrin deposition on the vascular graft material by inhibiting both the contact pathway and blood borne tissue factor procoagulant activity arising from either the alternatively spliced form of tissue factor or from tissue factor containing microparticles.     

Life expectancy in elderly patients following burns injury

Introduction

Burn injuries commonly occur in vulnerable age and social groups. Previous research has shown that frailty may represent a more important marker of adverse outcome in healthcare rather than chronological age (Roberts et al., 2012). In this paper we determined the relationship between burn injury, frailty, co-morbidities and long-term survival.

Effect of Surotomycin,a Novel Cyclic Lipopeptide Antibiotic,on Intestinal Colonization with Vancomycin-Resistant Enterococci and Klebsiella pneumoniae in Mice

Surotomycin (formerly called CB-183,315) is a novel, orally administered cyclic lipopeptide antibacterial in development for the treatment of Clostridium difficile infection (CDI) that has potent activity against vancomycin-resistant enterococci (VRE) but limited activity against Gram-negative bacilli, including Bacteroides spp. We used a mouse model to investigate the impact of surotomycin exposure on the microbiome, and to test the consequences of the disruption on colonization by vancomycin-resistant enterococci (VRE) and extended-spectrum β-lactamase-producing Klebsiella pneumoniae (ESBL-KP), in comparison with the effects of oral vancomycin and metronidazole. Mice (8 per group) received saline, vancomycin, metronidazole, or surotomycin through an orogastric tube daily for 5 days and were challenged with 10⁵ CFU of VRE or ESBL-KP administered through an orogastric tube on day 2 of treatment. The concentrations of the pathogens in stool were determined during and after treatment by plating on selective media. A second experiment was conducted to determine if the antibiotics would inhibit established VRE colonization. In comparison to controls, oral vancomycin promoted VRE and ESBL-KP overgrowth in stool (8 log₁₀ to 10 log₁₀ CFU/g; P < 0.001), whereas metronidazole did not (<4 log₁₀ CFU/g; P > 0.5). Surotomycin promoted ESBL-KP overgrowth (>8 log₁₀ CFU/g; P, <0.001 for comparison with saline controls) but not VRE overgrowth. Surotomycin suppressed preexisting VRE colonization, whereas metronidazole and vancomycin did not. These results suggest that treatment of CDI with surotomycin could reduce levels of VRE acquisition and overgrowth from those with agents such as vancomycin and metronidazole. However, surotomycin and vancomycin may promote colonization by antibiotic-resistant Gram-negative bacilli.     

cJun modulates Gγ-globin gene expression via an upstream cAMP response element

The upstream Gγ-globin gene cAMP response element (G-CRE) was previously shown to play a role in drug-mediated fetal hemoglobin induction. This effect is achieved via p38 mitogen activated protein kinase (MAPK)-dependent CREB1 and ATF-2 phosphorylation and G-CRE transactivation. Since this motif is also a predicted consensus binding site for cJun we extended our analysis to determine the ability of cJun to transactivate γ-globin through the G-CRE. Using chromatin immunoprecipitation assays we showed comparable in vivo cJun and CREB1 binding to the G-CRE region. Protein–protein interactions were confirmed between cJun/ATF-2 and CREB1/ATF-2 but not between CREB1 and cJun. However, we observed cJun and CREB1 binding to the G-CRE in vitro by electrophoretic mobility shift assay. Promoter pull-down assay followed by sequential western blot analysis confirmed co-localization of cJun, CREB1, and ATF-2 on the G-CRE. To show functional relevance, enforced expression studies with pLen-cJun and a Gγ-promoter (- 1500 to + 36) luciferase reporter were completed; we observed a concentration-dependent increase in luciferase activity with pLen-cJun similar to that produced by CREB1 enforced expression. Moreover, the G/A mutation at -1225 in the G-CRE abolished cJun transactivation. Finally, enforced cJun expression in K562 cells and normal primary erythroid progenitors enhanced endogenous γ-globin gene expression. We conclude that these data indicate that cJun activates the Gγ-globin promoter via the G-CRE in a manner comparable with CREB1 and propose a model for γ-globin activation based on DNA–protein interactions in the G-CRE.     

Increased power of mixed models facilitates association mapping of 10 loci for metabolic traits in an isolated population

The potential benefits of using population isolates in genetic mapping, such as reduced genetic, phenotypic and environmental heterogeneity, are offset by the challenges posed by the large amounts of direct and cryptic relatedness in these populations confounding basic assumptions of independence. We have evaluated four representative specialized methods for association testing in the presence of relatedness; (i) within-family (ii) within- and between-family and (iii) mixed-models methods, using simulated traits for 2906 subjects with known genome-wide genotype data from an extremely isolated population, the Island of Kosrae, Federated States of Micronesia. We report that mixed models optimally extract association information from such samples, demonstrating 88% power to rank the true variant as among the top 10 genome-wide with 56% achieving genome-wide significance, a >80% improvement over the other methods, and demonstrate that population isolates have similar power to non-isolate populations for observing variants of known effects. We then used the mixed-model method to reanalyze data for 17 published phenotypes relating to metabolic traits and electrocardiographic measures, along with another 8 previously unreported. We replicate nine genome-wide significant associations with known loci of plasma cholesterol, high-density lipoprotein, low-density lipoprotein, triglycerides, thyroid stimulating hormone, homocysteine, C-reactive protein and uric acid, with only one detected in the previous analysis of the same traits. Further, we leveraged shared identity-by-descent genetic segments in the region of the uric acid locus to fine-map the signal, refining the known locus by a factor of 4. Finally, we report a novel associations for height (rs17629022, P< 2.1 × 10??).     

Role of 14-bp insertion/deletion polymorphism in HLA-G among Indian women with recurrent spontaneous abortions

Human leukocyte antigen (HLA)-G is predominantly expressed on the extravillous cytotrophoblasts at the fetal-maternal interface. The 14-bp polymorphism in exon 8 is associated with HLA-G messenger ribonucleic acid (mRNA) stability and isoform alternative splicing patterns, thereby influencing the functionality of HLA-G in pregnancy. We analysed the 14-bp indel polymorphism in 143 recurrent spontaneous abortions (RSAs) and 150 control couples. We did not find any significant difference in the 14-bp insertion/deletion allele frequencies among the RSA and control couples. Analysis for increased sharing of the polymorphism in the RSA and the control couples also did not show any significant difference. However, we found an increase in the frequency of the 14-bp deletion homozygotes in the RSA women, which could lead to extremely high levels of soluble HLA-G (sHLA-G).     

Outcomes of Ahmed glaucoma valve implantation and risk factors for graft failure in eyes with Descemet’s stripping endothelial keratoplasty

International Ophthalmology - The aim of this study was to assess the efficacy of Ahmed glaucoma valve (AGV) in eyes with Descemet’s stripping endothelial keratoplasty (DSEK) and glaucoma and...