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BACKGROUND & AIMS: Anti-tumor necrosis factor (anti-TNF) therapies are the most widely used biologic drugs for treating immune-mediated diseases, but repeated administration can induce the formation of anti-drug antibodies. The ability to identify patients at increased risk for development of anti-drug antibodies would facilitate selection of therapy and use of preventative strategies.METHODS: We performed a genome-wide association study to identify variants associated with time to development of anti-drug antibodies in a discovery cohort of 1240 biologic-naïve patients with Crohn's disease starting infliximab or adalimumab therapy. Immunogenicity was defined as an anti-drug antibody titer ≥10 AU/mL using a drug-tolerant enzyme-linked immunosorbent assay. Significant association signals were confirmed in a replication cohort of 178 patients with inflammatory bowel disease.RESULTS: The HLA-DQA1*05 allele, carried by approximately 40% of Europeans, significantly increased the rate of immunogenicity (hazard ratio [HR], 1.90; 95% confidence interval [CI], 1.60-2.25; P = 5.88 × 10-13). The highest rates of immunogenicity, 92% at 1 year, were observed in patients treated with infliximab monotherapy who carried HLA-DQA1*05; conversely the lowest rates of immunogenicity, 10% at 1 year, were observed in patients treated with adalimumab combination therapy who did not carry HLA-DQA1*05. We confirmed this finding in the replication cohort (HR, 2.00; 95% CI, 1.35-2.98; P = 6.60 × 10-4). This association was consistent for patients treated with adalimumab (HR, 1.89; 95% CI, 1.32-2.70) or infliximab (HR, 1.92; 95% CI, 1.57-2.33), and for patients treated with anti-TNF therapy alone (HR, 1.75; 95% CI, 1.37-2.22) or in combination with an immunomodulator (HR, 2.01; 95% CI, 1.57-2.58).CONCLUSIONS: In an observational study, we found a genome-wide significant association between HLA-DQA1*05 and the development of antibodies against anti-TNF agents. A randomized controlled biomarker trial is required to determine whether pretreatment testing for HLA-DQA1*05 improves patient outcomes by helping physicians select anti-TNF and combination therapies. ClinicalTrials.gov ID: NCT03088449.

Objective: A previous study suggested that the frequency with which patients visit wound care clinics influences the rate of chronic wound healing but high bias was present with regard to wound care centre selection. The objective of this retrospective cohort study was to confirm this finding by using a very large sample size of diabetic foot ulcers (DFUs) from the US Wound Registry. Method: Patients who visited the clinic more than once for a new DFU were eligible for study inclusion. No exclusion was made with regard to Wagner grade, wound severity, or patient comorbidity. A Cox regression was conducted to analyse time to heal within 1 year using covariates known or suspected to influence wound healing, including visit frequency. Results: In terms of relative effect size, covariates that impeded wound healing the most were wound age and visit frequency with lower visit frequencies associated with lower probabilities of wound healing. Compared with DFUs (n=39,750) seen at a frequency of 7.5 times or more per 4 weeks, wounds seen at intervals of 2 weeks or less had a hazard ratio of 0.098 [(95% confidence interval: 0.09-0.11]. Using a separate breakpoint of ≥2 versus < 2 per 4 weeks specifically for the estimate of overall effect size, Cohen's w was 0.14—a small-to-medium effect size. Conclusion: Our findings confirm previous work and have implications for clinical practice and analysis of uncontrolled studies.

Over recent decades it has become increasingly apparent that malignant cells, including chronic lymphocytic leukemia (CLL) cells, do not exist in isolation. Rather they coalesce with numerous "normal" cells of the body and, in the case of CLL, inhabit key immunological niches within secondary lymphoid organs (SLO), where a plethora of stromal and immune cells mediate their growth and survival. With the advent and approval of targeted immune therapies such as monoclonal antibodies (mAb), which elicit their efficacy by engaging immune-mediated effector mechanisms, it is important to develop accurate methods to measure their activities. Here, we describe a series of reliable assays capable of measuring important antibody-mediated effector functions: antibody-dependent cellular phagocytosis (ADCP), antibody-dependent cellular cytotoxicity (ADCC), and complement-dependent cytotoxicity (CDC) that measure these immune activities.

Five species of Castanopsis namely C. armata, C. echinocarpa, C. hystrix, C. indica and C. tribuloides of Meghalaya were investigated to study the microscopic features and variation in their wood elements. It is observed that C. armata, C. hystrix and C. indica have semi ring porous wood with distinct growth rings while C. echinocarpa shows diffuse porous to semi ring porous wood with indistinct growth rings and C. tribuloides has semi ring porous to ring porous wood with distinct growth rings. Aggregate rays were observed only in C. armata. Both homocellular and heterocellular rays were present in all species. All Castanopsis species showed the presence of apotracheal diffuse, diffuse in aggregate, paratracheal scanty and vasicentric parenchyma. Tissue proportion measurements of different wood elements showed maximum percentage of fibers and minimum of parenchyma in all selected species. Mean minimum and maximum fiber length, fiber diameter, fiber wall thickness, vessel element length and vessel diameter were recorded as 1329.74 ± 9.03 (C. tribuloides)–1584.57 ± 7.79 μm (C. echinocarpa), 22.72 ± 2.76 (C. indica)–30.30 ± 7.89 μm (C. armata), 7.91 ± 1.77 (C. tribuloides)–8.95 ± 2.10 μm (C. hystrix), 722.28 ± 7.20 (C. tribuloides)–860.81 ± 13.52 μm (C. indica), 225.73 ± 29.49 (C. hystrix)–256 ± 31.06 μm (C. armata). Minimum and maximum vessel frequency were observed as 3.8 per mm2 (C. armata) and 5.2 per mm2 (C. tribuloides), while, minimum and maximum rays per mm were observed in C. hystrix and C. tribuloides. Variance ratio (F test) indicated the occurrence of significant differences among wood element dimensions except wall thickness of Castanopsis. Also, no correlation was observed between these wood elements except vessel frequency and wall thickness (r = −0.931*).

The main aim of present study is to gather original information on anatomical features and variations in dimensions of wood elements of some Quercus species of Meghalaya. All species of Quercus show diffuse porous wood with indistinct growth rings except Q. acutissima which shows tendency towards semi ring porous and Q. semiserrata with fairly distinct growth rings. Vessels are solitary or radial multiple of two arranged in radial or diagonal pattern. All species of Quercus show presence of scalariform perforation plates and vasicentric tracheids. Aggregate rays are present only in Q. acutissima where as Q. lanceofolia and Q. semiserrata show the presence of multiseriate rays. Both homocellular and heterocellular rays are present in Quercus species except Q. acutissima and Q. fenestrata which show only heterocellular rays. Axial parenchyma is apotracheal diffuse in all species except Q. acutissima that shows diffuse in aggregate, banded and unilateral paratracheal parenchyma in addition to apotracheal diffuse. Chambered crystals are present in Q. dealbata and Q. fenestrata.Tissue proportion of different wood elements shows that fiber percentage is maximum in all species. Variance ratio (F test) indicates that there are statistically significant differences in wood elements dimension namely fiber length, fiber diameter, vessel diameter, vessel density and wood density. Also a positive and significant correlation exists between fiber diameter and wall thickness while other parameters are not correlated with each other.

Ulcerative colitis (UC) and Crohn’s disease (CD) (collectively termed inflammatory bowel disease (IBD)) are complex disorders reflected by wide variation in clinical practice. These guidelines, commissioned by the Clinical Services’ Committee of the British Society of Gastroenterology (BSG) for clinicians and allied professionals caring for patients with IBD in the United Kingdom, provide an evidence based document describing good clinical practice for investigation and treatment. The guidelines are intended to bring consistency, but should not necessarily be regarded as the standard of care for all patients. Individual cases must be managed on the basis of all clinical data available for that case. Patient preferences should be sought and decisions jointly made between patient and health professional. ### 1.1 Development of guidelines A comprehensive literature search was performed using electronic databases (Medline, PubMed, and Ovid; keywords: “inflammatory bowel disease”, “ulcerative colitis”, and “Crohn’s disease”) by Dr Carter. A preliminary document was drafted by Dr Carter, Dr Lobo, and contributing authors. This was summarised by Dr Travis and revised after circulation first to the committee and then to members of the IBD section of the BSG, before submission to the Clinical Services’ Committee. ### 1.2 Grading of recommendations1 The guidelines conform to the North of England evidence based guidelines development project. The grading of each recommendation is dependent on the category of evidence supporting it: ### 1.3 Scheduled review of guidelines The content and evidence base …

Purpose: To determine whether metastasis-free survival following stereotactic radiosurgery is comparable with that following enucleation in a cohort of patients with choroidal and ciliary body melanoma. Methods: This was a non-randomized, retrospective study of 196 patients with uveal melanoma treated between 1990 and 2000. A total of 118 eyes were enucleated and 78 eyes were treated with stereotactic radiosurgery. The following variables were recorded: age and sex of patient; tumour location, volume and height; the presence of retinal detachment, and extrascleral extension. Tumour location was divided into three categories: within the posterior pole; beyond the limits of the posterior pole but not including the ciliary body, and ciliary body location. Analysis of survival time was performed using Kaplan−Meier estimation of survival curves and Cox's proportional hazards regression modelling. Results: The 5-year cumulative metastasis-free survival rate was 51% in the enucleation treatment group compared to 74% in the stereotactic treatment group. However, in the multivariate analysis there was no statistical difference in survival rates between the two treatment groups. The only variables that influenced survival rates were tumour location (p = 0.002), ciliary body tumours with the worst prognosis, and tumour volume (p = 0.001). Conclusions: Tumour size and location at presentation determined metastasis-free survival. Large ciliary body tumours had the highest risk of metastasis. Metastasis-free survival after stereotactic radiosurgery was comparable to that after enucleation.

Percutaneous endoscopic gastrostomy (PEG) is the accepted method for long-term enteral nutrition. Recent studies have suggested a higher mortality than was initially reported. The demands for gastrostomy insertion have risen, encompassing indications where the long-term outcomes are uncertain. We, therefore, constructed guidelines to try and improve the appropriateness of patient selection. Our aim was to prospectively assess the impact of guidelines for PEG insertion over a 1-yr period in a single center, Rotherham District General Hospital (hospital A) and compare against an adjacent center serving a similar population, Doncaster Royal Infirmary (hospital B) where guidelines had not been instituted.Data were collected from June, 1998 to May, 1999. Indication for PEG was documented and survival analysis performed using the Kaplan-Meier survival method.The number of PEG insertions had been rising each year in both centers. After guidelines were introduced in hospital A, the number of procedures fell by 20, whereas in hospital B, the PEG insertion rate continued to rise (p = 0.02). There was a lower mortality observed in hospital A (at 1 month, 16%; at 3 months, 26%; at 6 months, 39%; and at 1 yr, 46%) than in hospital B (at 1 month, 26%; at 3 months, 44%; at 6 months, 58%; and at I yr, 68%), although this did not achieve statistical significance (log rank test, p = 0.1).This is the first study to prospectively assess the impact of guidelines and a proactive role in the decision for PEG insertion. Fewer procedures were performed, and there was a trend toward a reduction in mortality.

A 50-year-old male with a 7 month history of progressive dysphagia to solids then subsequently to liquids. He underwent a diagnostic gastroscopy which was normal. A further barium swallow suggested achalasia. He was referred to a tertiary centre, where he underwent pH and manometry studies which confirmed a diagnosis of achalasia. He was referred for a laparoscopic cardiomyotomy, and at surgery there was a suspected tumour at the gastro-oesophageal junction. A follow-up endoscopy with biopsies was normal. Following this, a positron emission tomography scan showed T3 distal oesophageal cancer with no nodal involvement or distal metastasis. An attempt at oesophagectomy was performed, but at operation there was locally advanced carcinoma infiltrating the coeliac axis. He is currently undergoing palliative chemotherapy.

Summary Background Irritable bowel syndrome has a high prevalence. Consensus diagnostic criteria (ROME II) based on symptoms have been established to aid diagnosis. Although coeliac disease can be misdiagnosed as irritable bowel syndrome, no prospective study has been published in which patients with this disorder are investigated for coeliac disease. We aimed to assess the association of coeliac disease with irritable bowel syndrome in patients fulfilling ROME II criteria. Methods We undertook a case-control study at a university hospital. 300 consecutive new patients who fulfilled Rome II criteria for irritable bowel syndrome, and 300 healthy controls (age and sex matched) were investigated for coeliac disease by analysis of serum IgA antigliadin, IgG antigliadin, and endomysial antibodies (EMA). Patients and controls with positive antibody results were offered duodenal biopsy to confirm the possibility of coeliac disease. Findings 66 patients with irritable bowel syndrome had positive antibody results, of whom 14 had coeliac disease (11 EMA positive, three EMA negative). Nine patients with positive antibody results were lost to follow-up or refused biopsy (only one EMA-positive patient refused biopsy), and 43 had normal duodenal mucosa. Two controls, both of whom were EMA positive, had coeliac disease. Compared with matched controls, irritable bowel syndrome was significantly associated with coeliac disease (p=0·004, odds ratio=7·0 [95% CI 1·7–28·0]). Interpretation Patients with irritable bowel syndrome referred to secondary care should be investigated routinely for coeliac disease. With only EMA, three of 14 cases would have been missed.

Background & Aims: The interleukin 1 receptor antagonist gene allele 2 has been suggested as a determinant of both disease susceptibility and extent in ulcerative colitis. The aim of this study was to assess the allele as a predictor of both the indication for colectomy and the occurrence of pouchitis after ileal pouch–anal anastomosis formation. Methods: Genotyping for the +2018 single nucleotide polymorphism in the interleukin 1 receptor antagonist gene was performed in 109 patients who had undergone colectomy, including 82 patients who had been followed prospectively after ileal pouch–anal anastomosis formation. Results: Patients with pouchitis had a higher allele 2 carriage rate compared with those without pouchitis (72% vs. 45%) and Kaplan–Meier survival analysis showed that allele 2 carriers had a significantly increased incidence of pouchitis compared with noncarriers (log-rank test, 6.5). After adjustment for confounding covariates in a Cox proportional hazards model, the relative hazard was 3.1 (95% confidence interval [CI], 1.2–7.8; P = 0.02). Although there was a higher allele 2 carriage rate in patients with chronic refractory compared with acute severe ulcerative colitis (63% vs. 48%), this difference was not significant (odds ratio, 1.9; 95% Cl, 0.9–4.1; P = 0.1). Conclusions: The interleukin 1 receptor antagonist gene allele 2 predicts pouchitis after ileal pouch-anal anastomosis in ulcerative colitis. GASTROENTEROLOGY 2001;121:805-811

Background: Helicobacter pylori antimicrobial resistance is the most common reason for eradication failure. Small studies have shown metronidazole resistance to be more prevalent in certain population groups. Aim: To determine the resistance rates in a large cohort of patients from a single centre in the UK, and to evaluate resistance patterns over time, according to age, sex and socio-economic status. Methods: Consecutive patients with H. pylori-positive antral gastric biopsy samples were studied from 1994 to 1999. Susceptibility testing was performed to metronidazole, tetracycline, macrolide and amoxicillin by the modified disk diffusion method. The Jarman under-privileged area score was used as a measure of socio-economic status. Results: A total of 1064 patients were studied. Overall metronidazole resistance was 40.3%, decreasing with age (P

Percutaneous endoscopic gastrostomy (PEG) feeding has been validated in specific clinical situations such as acute stroke with dysphagia and oropharyngeal malignancy. The perception that gastrostomy insertion is safe and technically simple has led to an increase in the demands for PEG insertion, encompassing clinical applications such as in patients with dementia, in whom its role has not been justified. The purpose of this study was to compare the mortality of patients with dementia who were fed by PEG to that of other subgroups of patients requiring gastrostomy feeding.The study focused on a cohort of 361 consecutive patients requiring PEG feeding between August 1992 and July 1997 from two District General Hospitals (Rotherham District General Hospital and Doncaster Royal Infirmary) in South Yorkshire. A retrospective cohort survival analysis was performed using the Kaplan-Meier survival method and Cox proportional hazards analysis.In all patients requiring gastrostomy feeding there is a high initial mortality of 28% at 30 days. However, patients with dementia have a worse prognosis compared to other subgroups, with 54% having died at 1 month and 90% at 1 yr (log rank test p0.0001). This difference remained significant (log rank p0.0001) after adjusting for age at the time of PEG insertion.This is the first demonstration in the United Kingdom that the mortality of patients with dementia who are fed by gastrostomy is considerable. Consequently, we may wish to advise against gastrostomy feeding in selected patients within this clinical setting.

Single layers of Cu and In were deposited onto Mo coated glass substrates by radio frequency sputtering. The Cu11In9 phase has been found to be the majority phase in these precursors. The selenization of the sputtered layers has been achieved by depositing a 1 μm Se layer onto the precursor by thermal evaporation followed by an annealing in vacuum. Samples were annealed at different temperatures varying between 100–600°C at intervals of 50°C. The chalcopyrite structured ternary phase of CuInSe2 with a significant amount of preferential orientation in the (112) direction was obtained in samples annealed at 400°C or above. Morphological, compositional and structural analyses of the samples annealed at different temperatures were performed using a variety of complementary techniques. The results were analysed to explain the growth of CuInSe2 on the selenization of sputtered Cu-In precursors. The occurrence of various binary phases of Cu-In, Cu-Se and In-Se in different annealing temperature ranges has also been investigated. The phenomenon of volume expansion in CuInSe2 on selenization has been found to manifest itself as a shift in the characteristic (110) X-ray diffraction peak of Mo.

The standard method for back contacting to copper indium diselenide solar cells uses rf sputtered or e-beam evaporated molybdenum on a glass substrate. These processes are costly with a high energy input. A new simple technique has been proposed for the electroless deposition of a nickel - molybdenum alloy on chromium-coated glass substrates. This technique can also be applied directly to copper indium diselenide for a backwall cell configuration. Improved mechanical and electrical performances for this technique are reported.

The majority of complement protein C3 is synthesized by the liver, but many other cell types produce small amounts of functionally active C3. The overall contribution of such extrahepatic C3 production to the total circulating C3 level is unknown. Bone marrow and extrahepatic C3 productions were quantified in bone marrow transplant (BMT) and liver transplant (LT) recipients, respectively, where a mismatch for the C3 allotypes distinguished by the mAb HAV 4-1 had occurred. In the BMT group, donor-derived C3 was detected by ELISA and immunoblotting techniques. It contributed to 0.1 to 2.6% of the total circulating C3 during the immediate post-BMT period in response to inflammatory stimuli. Cell culture and immunostaining techniques demonstrated that monocytes were the source of the C3. By 6 wk following BMT, donor-derived C3 levels had decreased to below the detection limit of the assays. By contrast in the LT group, total extrahepatic C3 levels were higher (3.1-5.7% of the total circulating C3) and remained stable for up to 1 yr post-LT. This study demonstrates that extrahepatic derived C3 forms a larger proportion of the circulating C3 levels than was considered previously and that in the resting state, most of this extrahepatically derived C3 comes from nonmyeloid sources. In addition, monocytes, which in the resting state contribute negligible amounts of C3, have the potential when stimulated to contribute significantly to the total systemic C3 pool. These findings highlight the importance of locally secreted complement proteins.

Thin layers of In, Cu and Se were deposited at ambient temperature by thermal evaporation onto freshly cleaned glass substrates. The films were annealed in an inert atmosphere by an Argon-ion laser operating on all green lines with the power varied from 200 mW to 1 W. The films were characterized by scanning electron microscopy (SEM), energy dispersive X-ray analysis (EDAX) and X-ray diffractometry (XRD). The temperature-increment of the different layers under the incident laser powers were calculated theoretically. These results were used to suggest a mechanism for the formation of the CuInSe2 compound upon annealing the stack of elemental layers. It has been found that for the present form of the stack, the formation of the ternary chalcopyrite phase of CuInSe2 takes place through the appearance of the binary phase and subsequent reaction amongst them.

The genomic RNA of human astrovirus serotype 1 (HAst-1) contains three open reading frames (ORFs), 1a, 1b, and 2. ORF 1b is located downstream of, and overlaps, 1a, and it has been suggested on the basis of sequence analysis that expression of ORF 1b is mediated through -1 ribosomal frameshifting. To examine this possibility, a cDNA fragment containing the 1a-1b overlap region was cloned within a reporter gene and placed under the control of the bacteriophage SP6 promoter in a recombinant plasmid. Synthetic transcripts derived from this plasmid, when translated in the rabbit reticulocyte lysate cell-free system, specified the synthesis of polypeptides whose size and antibody reactivity were consistent with an efficient -1 ribosomal frameshift event at the overlap region. The HAst-1 frameshift signal has two essential components, a heptanucleotide slippery sequence, A6C, and a stem-loop structure in the RNA. The presence of this structure was confirmed by complementary and compensatory mutation analysis and by direct structure probing with single- and double-stranded RNA-specific reagents. The HAst-1 frameshift signal, like that present at the overlap of the gag and pro genes of the retrovirus human T-cell lymphotrophic virus type II, does not involve the formation of an RNA pseudoknot.

The optical and electrical properties of CulnSe2 (CIS) have shown it to be a promising photovoltaic material. The preparation of CulnSe2, thin films by means of the stacked elemental layer (SEL) technique was developed at Newcastle Photovoltaic Centre, Northumbria University. The method is presented as a low-cost technology because of its ability to retain stoichiometry and its scalability for large-area device applications. This paper presents a modification of the technique employed to produce CulnSe2 thin films and describes the structure, optical and electrical properties of such films. X-ray diffraction examination, energy dispersive x-ray analysis and measurements of the optical transmission and reflection and resistivity were used to determine the film properties relative to the preparation parameters and stoichiometry, The growth conditions were optimized for solar cell applications. Electrical resistivities in the range 10-2 to 105 Omega cm, energy gaps between 0.96 and 1.03 eV, and an absorption coefficient close to 105 for the CIS films were obtained in this work.

The solar cell structure is based upon a heterojunction of p-type CulnSe2 (CIS) and n-type CdS or mixed CdZnS. The use of a mixed CdZnS layer is reported to increase the open circuit voltage. The cells have been fabricated using the stacked elemental layer (SEL) technique for the fabrication of CIS thin films. The effect of layer resistivities on the performance of the device has been studied. The photoresponse was also determined and an essentially flat region between 0.6 and 1.0 mu m was found. Dark and light I-V characteristics in ambient and different solar cell temperatures have been investigated. Capacitance-voltage-frequency relationships were determined. A photovoltaic effect was observed with an ELH illumination (85 mW cm-2). Conversion efficiencies up to 5.2% were obtained. The results are discussed and recommendations for future work are suggested.

cDNA containing the entire coding sequence of the respiratory syncytial (RS) virus fusion (F) protein gene (574 amino acids) and two large PstI restriction fragments, encoding amino acids 18 to 212 and 214 to 574, were expressed in Escherichia coli as C-terminal chimeras with beta-galactosidase (beta-gal) in the pEX expression vector system. A further cDNA fragment, overlapping the PstI restriction site and encoding amino acids 190 to 289, was derived by PCR and expressed in a similar manner. Polyclonal rabbit serum raised against RS virus bound to all four chimeric proteins but most strongly to those containing C-terminal sequences. Two monoclonal antibodies (MAbs), 1E3 and RS348, capable of neutralizing the virus and inhibiting the viral fusion function, bound to all chimeras except that derived from the N-terminal PstI fragment, suggesting that their binding sites were located between amino acids 214 and 289. Further analysis of binding to expressed fragments from restriction enzyme digests and PCR amplification demonstrated that both antibodies bound to amino acids 253 to 289. MAb RS348 bound to 12-mer overlapping synthetic peptides containing the sequence 265 to 272 (PITNDQKK) but MAb 1E3 failed to bind to any 12-mer peptide derived from the F protein sequence. Immunization of mice with chimeric proteins containing the whole F protein coding sequence or amino acids 253 to 384, which includes the binding site of the two MAbs identified here, failed to induce antibodies that recognized the native RS virus F protein or could neutralize the virus. This suggests that either the beta-gal partner inhibits the immune response to the protein or that elements missing from the protein expressed in E. coli, perhaps conformational or added post-translation, contribute to the neutralizing antibody epitope.

We have determined the sequence for 1,000 bases from the 3' terminus of a human astrovirus serotype 1 isolated in Newcastle. This is the first sequence reported for a representative of this virus family. We find one open reading frame which terminates 83 bases from a poly A tail. The 3' non-coding-region has similarities to some picornavirus termini. However the amino acids specified by the coding region have no significant homology to the picornavirus protein 3D, encoded at the 3' terminus of these viruses. Northern blot analysis of intracellular virus-specific RNAs revealed one size of transcript which corresponded to full-length virus RNA. Available data thus indicate that astroviruses may resemble picornaviruses in replication strategy.

A one-step reverse transcription quantitative real-time polymerase chain reaction (RT-QPCR) method in combination with RNase treatment and low copy number samples was developed in order to examine the effect of temperature on the ability of virus capsids to protect their RNA content. The method was applied to a non-cultivable virus (GII.4 norovirus) and Feline calicivirus vaccine strain F-9 (FCV) which is often used as a norovirus surrogate. Results demonstrated that FCV RNA is exposed maximally after 2min at 63.3 degrees C and this correlated with a greater than 4.5log reduction in infectivity as assessed by plaque assay. In contrast human GII.4 norovirus RNA present in diluted clinical specimens was not exposed maximally until 76.6 degrees C, at least 13.3 degrees C greater than that for FCV. These data suggest that norovirus possesses greater thermostability than this commonly used surrogate. Further, these studies indicate that current food processing regimes for pasteurisation are insufficient to achieve inactivation of GII.4 NoVs. The method provides a novel molecular method for predicting virus infectivity.

Summary We report here the cloning and 3′ sequence determination of feline calicivirus strain F9. Subcloning the 3′ terminus enabled the production of strand specific probes for RNA synthesis. We extend the number of virus specific RNAs detected intracellularly to 8, and show that numbers 1–5 are represented as negative strands which may serve as templates in the synthesis of these RNAs.

We report results of a nonsynonymous SNP scan for ulcerative colitis and identify a previously unknown susceptibility locus at ECM1. We also show that several risk loci are common to ulcerative colitis and Crohn's disease (IL23R, IL12B, HLA, NKX2-3 and MST1), whereas autophagy genes ATG16L1 and IRGM, along with NOD2 (also known as CARD15), are specific for Crohn's disease. These data provide the first detailed illustration of the genetic relationship between these common inflammatory bowel diseases.

In this paper, we will present a new technique for fault isolation and failure analysis in integrated circuits based on a scanning magnetoresistive imaging system. By detecting the stray magnetic fields at the surface of a chip using magnetic sensors with sub-micron spatial resolution, we are able to obtain a full map of in-plane current densities, resolving features smaller than 100 nanometers. We will briefly discuss the capabilities and limitations of the technique and will present results on a variety of frontside and backside samples.

To determine whether metastasis-free survival following stereotactic radiosurgery is comparable with that following enucleation in a cohort of patients with choroidal and ciliary body melanoma.This was a non-randomized, retrospective study of 196 patients with uveal melanoma treated between 1990 and 2000. A total of 118 eyes were enucleated and 78 eyes were treated with stereotactic radiosurgery. The following variables were recorded: age and sex of patient; tumour location, volume and height; the presence of retinal detachment, and extrascleral extension. Tumour location was divided into three categories: within the posterior pole; beyond the limits of the posterior pole but not including the ciliary body, and ciliary body location. Analysis of survival time was performed using Kaplan-Meier estimation of survival curves and Cox's proportional hazards regression modelling.The 5-year cumulative metastasis-free survival rate was 51% in the enucleation treatment group compared to 74% in the stereotactic treatment group. However, in the multivariate analysis there was no statistical difference in survival rates between the two treatment groups. The only variables that influenced survival rates were tumour location (p = 0.002), ciliary body tumours with the worst prognosis, and tumour volume (p = 0.001).Tumour size and location at presentation determined metastasis-free survival. Large ciliary body tumours had the highest risk of metastasis. Metastasis-free survival after stereotactic radiosurgery was comparable to that after enucleation.

A novel method of producing CuInSe/sub 2/ thin films has been developed. This method has promising potential for low-cost, large-area production and could be easily transferred to industrial application. Preliminary results showed that CuInSe/sub 2/ films are homogeneous over the substrate area used ( approximately 50 cm/sup -2/). All the cells produced from the same film demonstrated efficiencies of around 3%. The highest cell efficiency obtained was 3.5%. The corresponding device parameters were V/sub oc/=260 mV, J/sub sc/=25 mA/cm/sup 2/, and FF=45% under ELH illumination, 85 mW/cm/sup 2/. Results from capacitance-voltage and quantum efficiency measurements are also discussed. >

BACKGROUND AND AIMS—An association between the allele 2 of the interleukin 1 receptor antagonist gene variable number tandem repeats polymorphism in intron 2 and ulcerative colitis was first reported in 1994. Subsequent studies in Caucasian Northern European patients have not confirmed this, although trends towards an association were observed. The lack of statistical significance could reflect inadequate power. In this study the association was reassessed in a large independent set of well characterised Caucasian patients and a meta-analysis of reported patient series was performed. PATIENTS AND METHODS—A total of 320 patients with endoscopically and histologically confirmed ulcerative colitis (124 pancolitis, 196 left sided and distal disease) and 827 ethnically matched controls were genotyped at polymorphic sites in the interleukin 1 receptor antagonist gene. Carriage rates were compared using χ2 statistics. A meta-analysis of this and seven previous studies in North European Caucasian patients was performed using the Mantel-Haenszel χ2 test. RESULTS—Patients had a significantly increased carriage rate of allele 2 compared with controls (52% v 45%; odds ratio 1.3 (95% confidence interval (CI) 1.01-1.7); p=0.04). The allele 2 carriage rate was highest in extensive colitis (carriage rate 56%; odds ratio 1.5 (95% CI 1.1-2.3) p=0.02) and in individuals who had undergone colectomy (carriage rate 55%; odds ratio 1.5 (95% CI 0.95-2.4); p=0.08). Meta-analysis of all eight studies showed a significant association between carriage of allele 2 and ulcerative colitis (odds ratio 1.23 (95% CI 1.04-1.45); p=0.01). CONCLUSIONS—The association of the interleukin 1 receptor antagonist gene polymorphism with ulcerative colitis is confirmed. The association is minor and confers only a small risk to an individual but will contribute a high attributable risk in a population due to the high allelic frequency. Accurate phenotypic characterisation defines more homogeneous subsets of patients, such as those with extensive disease, in whom the association is greater. Keywords: ulcerative colitis; cytokine gene polymorphisms; interleukin 1 receptor antagonist; interleukin 1; inflammatory bowel disease; genetics

6-Mercaptopurine and azathioprine are established treatments for Crohn's disease, both for maintenance1 and induction of remission.2 They are the agents of choice for steroid dependant disease. Their use in Crohn's disease is limited in two ways. Firstly, potentially serious dose dependant and idiosyncratic hypersensitivity side effects occur in a significant minority of patients treated. Secondly, the clinical effect has been said to have a prolonged time to onset with response rates only significantly increasing after 17 weeks of therapy,2 limiting usefulness in refractory active disease. Pharmacokinetic studies have suggested that the active metabolites of these drugs, the 6-thioguanine nucleotides (6TGN), have a long half life in red blood cells and slowly accumulate in tissues, requiring prolonged treatment to achieve steady state levels.3 Sandborn and colleagues have carefully addressed whether an intravenous loading dose of azathioprine would reduce the time to respond in patients with …

Percutaneous endoscopic gastrostomy (PEG) is the commonest method of long-term enteral nutrition. In the UK there is no nationally funded framework to provide community support for patients with gastrostomies.To assess the hospital support required and the complications occurring in a cohort of patients discharged into the community with a gastrostomy in-situ.We prospectively collected data between June and November 1998 regarding patients (n=87) with PEG complications in the community following discharge from two adjacent hospitals in South Yorkshire (Rotherham District General Hospital and the Royal Hallamshire Hospital).The mortality within this cohort during the study period was 17.2%. Of these 15 patients 10.3% (n=9) were thought to have died as a result of the progression of their disease, whilst in the other 6.9% (n=6) pneumonia was documented as the cause of death. Problems with gastrostomies requiring telephone advice occurred in 24.1% of patients and 65.5% necessitated a home visit. However during this period of 6 months, 23% of patients were admitted as an emergency, accounting for 61 in-patient days. All were from nursing or residential homes and the emergency occurred after normal surgery hours. General practitioners dealt with aspiration pneumonia in 25.2% of the cohort but other tube problems were referred to the endoscopy unit.This is the first study to specifically highlight the specialist intervention and support required by patients discharged into the community with PEGs, a group for whom there is often no formal aftercare. A specialist nurse or dietician could establish a liaison service focusing on primary care and using hospital resources when appropriate. This study provides further evidence to support the recommendations for tube feeding made by the British Association for Parenteral and Enteral Nutrition.

We have determined the nucleotide sequence of sacbrood virus (SBV), which causes a fatal infection of honey bee larvae. The genomic RNA of SBV is longer than that of typical mammalian picornaviruses (8832 nucleotides) and contains a single, large open reading frame (179-8752) encoding a polyprotein of 2858 amino acids. Sequence comparison with other virus polyproteins revealed regions of similarity to characterized helicase, protease and RNA-dependent RNA polymerase domains; structural genes were located at the 5' terminus with non-structural genes at the 3' end. Picornavirus-like agents of insects have two distinct genomic organizations; some resemble mammalian picornaviruses with structural genes at the 5' end and non-structural genes at the 3' end, and others resemble caliciviruses in which this order is reversed; SBV thus belongs to the former type. Sequence comparison suggested that SBV is distantly related to infectious flacherie virus (IFV) of the silk worm, which possesses an RNA of similar size and gene order.

During 1997, an extensive outbreak of astrovirus occurred in a unit where paediatric patients were being treated for leukaemias and inherited immune deficiency disorders. Prolonged shedding of virus for many months following infection was demonstrated in three patients who had undergone bone marrow transplantation. Comparison of reverse transcription-polymerase chain reaction (RT-PCR), enzyme immunoassay (EIA), and electronmicroscopy (EM) to monitor the outbreak showed that many subclinical infections, mainly in children aged3 years could only be detected by RT-PCR. Use of RT-PCR revealed that several patients were infected earlier and shed virus for longer than by using EM or EIA. The virus responsible for the outbreak was identified as HAstV-1 and was shown to have a sequence that differed from a strain obtained in 1988.

Assessment of oestrogen and progesterone receptors (ER and PgR) in breast cancer is widely used for the prediction of response to endocrine therapy and as a prognostic marker. Cytosolic assays have been replaced in many centres by immunochemical techniques, which have many advantages including applicability to small samples, simplicity, and cost-effectiveness. This study describes the generation and characterisation of two novel murine monoclonal antibodies recognizing ER and PgR, designated NCL-ER-6F11 and NCL-PGR respectively, which are effective in heat-treated formalin-fixed, paraffin-embedded tissue. The antibodies have been characterized by Western blotting and by immunohistochemistry on normal and pathological breast and other tissues. NCL-ER-6F11 has been shown to compare favourably with a currently available ER antibody. These antibodies may prove of value in the assessment of hormone receptor status in human breast cancer.

Astroviruses (genus Astrovirus) are assigned to a newly established virus family, the Astroviridae. The molecular biology of these agents reveals many features unique amongst the non-enveloped animal viruses and resembles that of members of certain plant virus families. In particular, their possession of a serine protease and use of ribosomal frameshifting to express the RNA polymerase are similar to the luteoviruses. Many aspects of the astrovirus replication strategy are still unclear, but replication may involve a nuclear step and non-structural proteins may influence host cell range.

Astroviruses (genus Astrovirus) are assigned to a newly established virus family, the Astroviridae. The molecular biology of these agents reveals many features unique amongst the non-enveloped animal viruses and resembles that of members of certain plant virus families. In particular, their possession of a serine protease and use of ribosomal frameshifting to express the RNA polymerase are similar to the luteoviruses. Many aspects of the astrovirus replication strategy are still unclear, but replication may involve a nuclear step and non-structural proteins may influence host cell range.

Caffeine is a commonly used drug that increases arousal, a condition associated with increased cholinergic activity in the mammalian cerebral cortex including the hippocampus. We have used the technique of microdialysis in association with microbore high-performance liquid chromatography to investigate the effects of caffeine on the extracellular levels of acetylcholine in the hippocampus of awake, freely moving rats. The oral administration of caffeine dose-dependently (3-30 mg/kg) increased the extracellular levels of acetylcholine. This increase was completely blocked when the microdialysis probe was perfused with the Na(+)-channel blocker tetrodotoxin, and strongly attenuated when a Ca(2+)-free Ringer solution was used. The effect of caffeine on hippocampal acetylcholine release was concentration-dependently counteracted by local perfusion of an A1 receptor agonist, N6-cyclopentyladenosine (0.1-1 mumol/liter), but not by the A2 receptor agonist, CGS 21680 (10 mumol/liter). Neither agonist affected base-line acetylcholine release at these concentrations. These results demonstrate that acetylcholine release in the hippocampus is under tonic inhibitory control of the endogenous neuromodulator adenosine, and that orally administered caffeine enhances action potential-dependent vesicular acetylcholine release by antagonism of local A1 receptors. Hence, the data provide a possible link between adenosine A1 receptors in the hippocampus, increased cholinergic activity and the psychostimulant effects of caffeine.

The neutralizing activity and fusion-inhibition activity per unit weight of immunoglobulin were determined for each of a panel of 20 monoclonal antibodies (MAbs) to the fusion (F) protein of respiratory syncytial (RS) virus. Neutralization did not correlate with fusion-inhibiting activity, suggesting that the F protein plays at least two independent, antibody-sensitive roles in viral infection. Antibodies with the highest biological activity against A2, a subgroup A strain of RS virus, neutralized a subgroup B strain (8/60) poorly, suggesting a degree of antigenic variation that may be important in human infection. All but one fusion-inhibiting MAb bound to protein blots and binding was mapped to two areas on overlapping F protein fragments. One MAb with relatively poor fusion-inhibiting activity bound only to fragments C-terminal of amino acid 384, the remainder bound only to fragments containing residues 253 to 289. MAbs directed to the latter site were heterogeneous in neutralizing activity, subgroup specificity and fusion-inhibiting activity. These variations between MAbs could not be accounted for by differences in their binding avidities. We suggest that this binding site is not the complete antibody epitope which probably includes conformation-dependent elements.

Astroviruses and caliciviruses are two families defined initially by their characteristic morphology. Many of these viruses have been difficult to grow in culture. Molecular biology has now provided a valuable insight into the nature of these viruses, and in many respects knowledge of genome structure now outstrips that of more classical virological features. However these advances have allowed a more detailed approach to virus classification and have led to the establishment of the Astroviridae as a distinct virus family.

Astroviruses and caliciviruses are two families defined initially by their characteristic morphology. Many of these viruses have been difficult to grow in culture. Molecular biology has now provided a valuable insight into the nature of these viruses, and in many respects knowledge of genome structure now outstrips that of more classical virological features. However these advances have allowed a more detailed approach to virus classification and have led to the establishment of the Astroviridae as a distinct virus family.

Editor,—The presentation of abstracts at scientific meetings provides an opportunity to rapidly convey the results of novel research. It also allows the researcher a chance to receive informal peer review. This may help to clarify aspects of the work, particularly in the identification and correction of potential weaknesses prior to submission for full publication. Although abstracts submitted to conferences are peer reviewed, this process may not be as rigorous as that of an indexed journal considering publication of the full manuscript.1 Presentation of an abstract at a prestigious meeting may suggest that full publication is probable. Certainly, acceptance as opposed to rejection increases the likelihood of subsequent publication, but this is not absolute.2 Other medical specialities …

tion rates and multiple choice question (MCQ) scores (r=0.24 and 0.27, p

Introduction The demand for endoscopy is ever increasing as is the rise in overall costs of endoscopy and waiting times. Adherence to appropriate indications is essential to prevent the rise in proportions of negative and hence unnecessary endoscopies. Patients with chronic renal impairment are one group of patients who may have significant gastrointestinal symptoms and often form an important number of referrals to the endoscopy unit for upper GI endoscopy. These patients are of variable complexity and often have multi-organ disease. Anaemia can often be the indication for referral despite this being part of the disease complex. We wanted to investigate the indications and endoscopic findings in renal patients from a busy tertiary referral centre. Methods The aim was to assess the findings at gastroscopy in patients with renal failure from a renal tertiary referral centre. This was a single centre, retrospective analysis of renal patients who were referred for gastroscopy. Patient details were obtained from the CIPTS database by isolating patients under the named renal physician referrer. Gastroscopy reports between 2006 until April 2009 were obtained and analysed for indications and diagnosis at endoscopy. Results A total of 90 gastroscopies in 73 patients were conducted during the study period. The age range was 45–92 and 58% were male patients. 86% of the gastroscopies were inpatient procedures. The majority of indications for gastroscopies included anaemia, haematemesis or melaena and these accounted for the indication 75 times either alone or in combination. Out of 90 gastroscopies 31 (34%) were considered as normal. Those gastroscopies in the normal category included reports of normal gastroscopies, hiatus hernias or minimal gastritis. All else was classed as abnormal (66%). Erosive lesions within the upper GI tract were diagnosed 47 times in the 90 reports (52%). These included oesophageal, gastric and duodenal ulcers, mallory weiss tears, duodenal erosions, and pyloric and duodenal stenoses. Three reports had cancer (1 duodenal, 2 oesophageal), and vascular lesions (angiodysplasia, telangiectasia, portal gastropathy, varices) were reported 14 (16%) times. Conclusion Two-thirds of patients with renal failure referred for an upper GI endoscopy have abnormal findings. The majority have erosive disorders as opposed to vascular lesions as previously reported to be associated in the literature. Our study demonstrates a high pick-up rate of in-patients with renal failure and we recommend that the threshold to endoscopy in such patients should be low, and anaemia should not be ascribed to one of chronic disease without performing a gastroscopy.

This paper presents recent results on the study of the Stacked Elemental Layer (SEL) technique developed at Newcastle Polytechnic for the fabrication of CuInSe2 thin films onto large area substrates as a low cost technology. The method has now been developed to produce solar cells using either Cd(Zn)S or indium doped CdS using standard high vacuum evaporation for the window layer deposition. It has been found that well structured chalcopyrite CuInSe2 with the desired electro-optical properties can be formed. The CuInSe2 films produced exhibit resistivity variations with composition comparable to those produced by more conventional techniques and crystallinity, as demonstrated by X-ray diffraction measurements, exceeding some such conventional films. Film properties as a function of S.E.L. deposition parameters have been fully delineated. To date device efficiencies approaching 4% under 85 mW cm-2 ELH illumination have been demonstrated. Some salient points highlighting the potential of the SEL technique for commercial application are made.

A retrospective study was carried out to determine the results of laser treatment for choroidal neovascular membranes in age-related macular degeneration in 92 patients in whom fluorescein angiography was performed for this condition over a 7-year period. Twenty-nine of these patients, treated with the argon laser, were followed up regularly for 15 months. The treatment comprised overlapping laser spots of 200 microns and an average power of 390 mW. The results of this study demonstrated that: (i) the majority of membranes were closer than 500 microns from the foveola; (ii) the second eye involvement rate in the same patient was 15,79% over 15 months; and (iii) a 37,5% significant visual loss or 41% two lines or greater visual loss was seen after 15 months, which compares favourably with results of laser treatment reported by other institutions.