eNOS overexpression exacerbates vascular closure in the obliterative phase of OIR and increases angiogenic drive in the subsequent proliferative stage

Purpose: In ischemic retinopathies, the misdirection of reparative angiogenesis away from the hypoxic retina leads to pathologic neovascularization. Thus, therapeutic strategies that reverse this trend would be extremely beneficial. Nitric oxide (NO) produced by endothelial nitric oxide synthase (eNOS) is an important mediator of vascular endothelial growth factor (VEGF) function facilitating vascular growth and maturation. However, in addition to NO, eNOS can also produce superoxide (O), exacerbating pathology. Here, our aim was to investigate the effect of eNOS overexpression on vascular closure and subsequent recovery of the ischemic retina.

Methods: Mice overexpressing eNOS-GFP were subjected to oxygen-induced retinopathy (OIR) and changes in retinal vascularization quantified. Background angiogenic drive was assessed during vascular development and in aortic rings. NOS activity was measured by Griess assay or conversion of radiolabeled arginine to citrulline, nitrotyrosine (NT), and superoxide by immunolabeling and dihydroethidium fluorescence and VEGF by ELISA.

Results: In response to hyperoxia, enhanced eNOS expression led to increased NOS-derived superoxide and dysfunctional NO production, NT accumulation, and exacerbated vessel closure associated with tetrahydrobiopterin (BH) insufficiency. Despite worse vaso-obliteration, eNOS overexpression resulted in elevated hypoxia-induced angiogenic drive, independent of VEGF production. This correlated with increased vascular branching similar to that observed in isolated aortas and during development. Enhanced recovery was also associated with neovascular tuft formation, which showed defective NO production and increased eNOS-derived superoxide and NT levels.

Conclusions: In hyperoxia, reduced BH bioavailability causes overexpressed eNOS to become dysfunctional, exacerbating vaso-obliteration. In the proliferative phase, however, eNOS has important prorepair functions enhancing angiogenic growth potential and recovery in ischemia. © 2012 The Association for Research in Vision and Ophthalmology, Inc.

Consensus statements for management of Barrett's dysplasia and early-stage esophageal adenocarcinoma, based on a Delphi process

Esophageal adenocarcinoma (EA) is increasingly common among patients with Barrett's esophagus (BE). We aimed to provide consensus recommendations based on the medical literature that clinicians could use to manage patients with BE and low-grade dysplasia, high-grade dysplasia (HGD), or early-stage EA.

Allele resolution of HLA-A using oligonucleotide probes in a two-stage typing strategy

High-resolution polymerase chain reaction using sequence-specific oligonucleotide probes (PCR-SSOP) typing methods for HLA-A identification have been established. The four systems, which operate independently of each other, are intended for use as secondary typing systems following HLA-A identification with a medium-resolution PCR-SSOP technique. The systems, all using digoxigenin-labelled probes, are based on group specific amplifications for resolution of: i) HLA-A*29 & -A*33; ii) HLA-A*24 & -A*30; and iii) HLA-A*26, -A*25, -A*11, -A*34, -A*66 and -A*68 alleles, respectively. The fourth system, for the detection of HLA-A*02 alleles, is a modification of a previously reported PCR-SSOP subtyping system. The methods have been applied to individuals from the local bone marrow registry and HLA-A allele frequencies for the Northern Ireland population have been established.

Kruppel-associated Box (KRAB)-associated co-repressor (KAP-1) Ser-473 phosphorylation regulates heterochromatin protein 1β (HP1-β) mobilization and DNA repair in heterochromatin

The DNA damage response encompasses a complex series of signaling pathways that function to regulate and facilitate the repair of damaged DNA. Recent studies have shown that the repair of transcriptionally inactive chromatin, named heterochromatin, is dependent upon the phosphorylation of the co-repressor, Krüppel-associated box (KRAB) domain-associated protein (KAP-1), by the ataxia telangiectasia-mutated (ATM) kinase. Co-repressors, such as KAP-1, function to regulate the rigid structure of heterochromatin by recruiting histone-modifying enzymes, such HDAC1/2, SETDB1, and nucleosome-remodeling complexes such as CHD3. Here, we have characterized a phosphorylation site in the HP1-binding domain of KAP-1, Ser-473, which is phosphorylated by the cell cycle checkpoint kinase Chk2. Expression of a nonphosphorylatable S473A mutant conferred cellular sensitivity to DNA-damaging agents and led to defective repair of DNA double-strand breaks in heterochromatin. In addition, cells expressing S473A also displayed defective mobilization of the HP1-ß chromodomain protein. The DNA repair defect observed in cells expressing S473A was alleviated by depletion of HP1-ß, suggesting that phosphorylation of KAP-1 on Ser-473 promotes the mobilization of HP1-ß from heterochromatin and subsequent DNA repair. These results suggest a novel mechanism of KAP-1-mediated chromatin restructuring via Chk2-regulated HP1-ß exchange from heterochromatin, promoting DNA repair.

Performing History on the Victorian Stage

An investigation of the use of historically accurate scenery in Victorian revivals of Shakespeare's history plays.

Impact damage and repair of composite structures

This paper gives an overview of the work carried out in a GARTEUR (Group for Aeronautical Research and Technology in Europe) program, under the chairmanship of the author, to develop and validate analytical and numerical methods to characterise real impact damage in composite structures, particularly those designed to sustain load in a postbuckled state, and to study the durability of bonded repairs. GARTEUR is an inter-governmental agreement between the seven European countries with the largest direct employment in the Aerospace industry, to mobilise scientific and technical knowledge between the member countries. A number of Action Groups have been launched, since GARTEUR’s inception in the early 1970s, to address specific technical issues of interest to the participating members. The research presented in this paper was performed under Action Group 28 with partners from ONERA, EADS-CCR (France), DLR, AIRBUS-Deutschland, EADS-M (Germany), CIRA (Italy), INTA (Spain), SICOMP, Saab, (Sweden), NLR (The Netherlands), QinetiQ, BAE Systems, Imperial College London and the University of Sheffield (United Kingdom). The Action Group tasks were divided into four Work Elements (WEs): WE1-Prediction and characterisation of impact damage, WE2- Postbuckling with delamination, WE3-Repair and WE4-Fatigue. This paper outlines the main developments and achievements within each Work Element.

A late-Middle Pleistocene (Marine Isotope Stage 6) vegetated surface buried by Old Crow tephra at the Palisades, interior Alaska

A 40 cm thick primary bed of Old Crow tephra (131 ± 11 ka), an important stratigraphic marker in eastern Beringia, directly overlies a vegetated surface at Palisades West, on the Yukon River in central Alaska. Analyses of insect, bryophyte, and vascular plant macrofossils from the buried surface and underlying organic-rich silt suggest the local presence of an aquatic environment and mesic shrub-tundra at the time of tephra deposition. Autochthonous plant and insect macrofossils from peat directly overlying Old Crow tephra suggest similar aquatic habitats and hydric to mesic tundra environments, though pollen counts indicate a substantial herbaceous component to the regional tundra vegetation. Trace amounts of arboreal pollen in sediments associated with the tephra probably reflect reworking from older deposits, rather than the local presence of trees. The revised glass fission-track age for Old Crow tephra places its deposition closer to the time of the last interglaciation than earlier age determinations, but stratigraphy and paleoecology of sites with Old Crow tephra indicate a late Marine Isotope Stage 6 age. Regional permafrost degradation and associated thaw slumping are responsible for the close stratigraphic and paleoecological relations between Old Crow tephra and last interglacial deposits at some sites in eastern Beringia. © 2009 Elsevier Ltd.

An Experimental and Numerical Study of the Static and Fatigue Performance of a Composite Adhesive Repair

Experimental static and fatigue tension-tension tests were carried out on 5HS/RTM6 composite intact coupons and coupons incorporating adhesively-bonded (FM300-2) stepped flush joints. The results show that the adhesive joint, which is widely used in repairs, significantly reduces the static strength as well as the fatigue life of the composite. Both, the static and the fatigue failure of the ‘repaired’ coupons occur at the adhesive joint and involve crack initiation and propagation. The latter is modelled using interface finite elements based on the decohezive zone approach. The material degradation in the interface constitutive law is described by a damage variable, which can evolve due to the applied loads as well as the number of fatigue cycles. The fatigue formulation, based on a published model, is adapted to fit the framework of the pseudotransient formulation that is used as a numerical tool to overcome convergence difficulties. The fatigue model requires three material parameters. Numerical tests show that a single set of these parameters can be used to recover, very accurately, the experimental S-N relationship. Sensitivity studies show that the results are not mesh dependent.

Availability of stage at diagnosis, cancer treatment delay and compliance with cancer guidelines as cancer registry indicators for cancer care in Europe:Results of EUROCHIP-3 survey

EUROCHIP (European Cancer Health Indicators Project) focuses on understanding inequalities in the cancer burden, care and survival by the indicators "stage at diagnosis," "cancer treatment delay" and "compliance with cancer guidelines" as the most important indicators. Our study aims at providing insight in whether cancer registries collect well-defined variables to determine these indicators in a comparative way. Eighty-six general European population-based cancer registries (PBCR) from 32 countries responded to the questionnaire, which was developed by EUROCHIP in collaboration with ENCR (European Network of Cancer Registries) and EUROCOURSE. Only 15% of all the PBCR in EU had all three indicators available. The indicator "stage at diagnosis" was gathered for at least one cancer site by 81% (using TNM in 39%). Variables for the indicator "cancer treatment delay" were collected by 37%. Availability of type of treatment (30%), surgery date (36%), starting date of radiotherapy (26%) and starting date of chemotherapy (23%) resulted in 15% of the PBCRs to be able to gather the indicator "compliance to guidelines". Lack of data source access and qualified staff were the major reasons for not collecting all the variables. In conclusion, based on self-reporting, a few of the participating PBCRs had data available which could be used for clinical audits, evaluation of cancer care projects, survival and for monitoring national cancer control strategies. Extra efforts should be made to improve this very efficient tool to compare cancer burden and the effects of the national cancer plans over Europe and to learn from each other. © 2012 UICC.

Hepatocellular carcinoma and polymorphisms in carcinogen-metabolizing and DNA repair enzymes in a population with aflatoxin exposure and hepatitis B virus endemicity

High rates of hepatocellular carcinoma (HCC) in The Gambia, West Africa, are primarily due to a high prevalence of chronic hepatitis B virus infection and heavy aflatoxin exposure via groundnut consumption. We investigated genetic polymorphisms in carcinogen-metabolizing (GSTM1, GSTT1, HYL1*2) and DNA repair (XRCC1) enzymes in a hospital-based case-control study. Incident HCC cases (n = 216) were compared with frequency-matched controls (n = 408) with no clinically apparent liver disease. Although the prevalence of variant genotypes was generally low, in multivariable analysis (adjusting for demographic factors, hepatitis B virus, hepatitis C virus, and TP53 status), the GSTM1-null genotype [odds ratio (OR), 2.45; 95% confidence interval (95% CI), 1.21-4.95] and the heterozygote XRCC1-399 AG genotype (OR, 3.18; 95% CI, 1.35-7.51) were significantly associated with HCC. A weak association of the HYL1*2 polymorphism with HCC was observed but did not reach statistical significance. GSTT1 was not associated with HCC. The risk for HCC with null GSTM1 was most prominent among those with the highest groundnut consumption (OR, 4.67; 95% CI, 1.45-15.1) and was not evident among those with less than the mean groundnut intake (OR, 0.64; 95% Cl, 0.20-2.02). Among participants who had all three suspected aflatoxin-related high-risk genotypes [GSTM1 null, HLY1*2 (HY/HH), and XRCC1 (AG/GG)], a significant 15-fold increased risk of HCC was observed albeit with imprecise estimates (OR, 14.7; 95% CI, 1.27-169). Our findings suggest that genetic modulation of carcinogen metabolism and DNA repair can alter susceptibility to HCC and that these effects may be modified by environmental factors.