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Background The pH of the human gastric mucosa varies around 2.5 so that only bacteria with strong acidic stress tolerance can colonize it. The ulcer causing Helicobacter pylori thrives in the gastric mucosa. We analyse the roles of the key outer membrane protein OMPLA in its roles in acid tolerance. Results The homology model of Helicobacter pylori outer membrane phospholipase A (OMPLA) reveals a twelve stranded [beta]-barrel with a pore that allows molecules to pass with a diameter up to 4 Ã. Structure based multiple sequence alignments revealed the functional roles of many amino acids, and led to the suggestion that OMPLA has multiple functions. Besides its role as phospholipase it lets urea enter and ammonium exit the periplasm. Combined with an extensive literature study, our work leads to a comprehensive model for H. pylori's acid tolerance. This model is based on the conversion of urea into ammonium, and it includes multiple roles for OMPLA and involves two hitherto little studied membrane channels in the OMPLA operon. Conclusion The three-dimensional model of OMPLA predicts a transmembrane pore that can aid H. pylori's acid tolerance through urea influx and ammonium efflux. After urea passes through OMPLA into the periplasm, it passes through the pH-gated inner membrane channel UreI into the cytoplasm where urease hydrolyses it into NH.sub.3 and CO.sub.2. Most of the NH.sub.3 becomes NH.sub.4.sup.+ that is likely to need an inner membrane channel to reach the periplasm. Two genes that are co-regulated with OMPLA in gastric Helicobacter operons could aid this transport. The NH.sub.4.sup.+ that might leave the cell through the OMPLA pore has been implicated in H. pylor's pathogenesis. Keywords: Helicobacter pylori, Acid tolerance, Multifunctional OMPLA, Urea pathway, Author(s): Hilde S. Vollan[sup.1,2] , Tone Tannaes[sup.1] , Dominique A. Caugant[sup.2,3] , Gert Vriend[sup.4] and Geir Bukholm[sup.2,5] Background H. pylori survives in the human gastric mucosa Most bacterial proteins require [...]

Abstract Background In the past decade, researchers have proposed that the pldA gene for outer membrane phospholipase A (OMPLA) is important for bacterial colonization of the human gastric ventricle. Several conserved Helicobacter pylori genes have distinct genotypes in different parts of the world, biogeographic patterns that can be analyzed through phylogenetic trees. The current study will shed light on the importance of the pldA gene in H. pylori. In silico sequence analysis will be used to investigate whether the bacteria are in the process of preserving, optimizing, or rejecting the pldA gene. The pldA gene will be phylogenetically compared to other housekeeping (HK) genes, and a possible origin via horizontal gene transfer (HGT) will be evaluated through both intra- and inter-species evolutionary analyses. Results In this study, pldA gene sequences were phylogenetically analyzed and compared with a large reference set of concatenated HK gene sequences. A total of 246 pldA nucleotide sequences were used; 207 were from Norwegian isolates, 20 were from Korean isolates, and 19 were from the NCBI database. Best-fit evolutionary models were determined with MEGA5 ModelTest for the pldA (K80 + I + G) and HK (GTR + I + G) sequences, and maximum likelihood trees were constructed. Both HK and pldA genes showed biogeographic clustering. Horizontal gene transfer was inferred based on significantly different GC contents, the codon adaptation index, and a phylogenetic conflict between a tree of OMPLA protein sequences representing 171 species and a tree of the AtpA HK protein for 169 species. Although a vast majority of the residues in OMPLA were predicted to be under purifying selection, sites undergoing positive selection were also found. Conclusions Our findings indicate that the pldA gene could have been more recently acquired than seven of the HK genes found in H. pylori. However, the common biogeographic patterns of both the HK and pldA sequences indicated that the transfer occurred long ago. Our results indicate that the bacterium is preserving the function of OMPLA, although some sites are still being evolutionarily optimized.

Abstract Background The association between Helicobacter pylori infection and upper gastrointestinal disease is well established. However, only a small fraction of H. pylori carriers develop disease, and there are great geographical differences in disease penetrance. The explanation to this enigma lies in the interaction between the bacterium and the host. H. pylori Outer Membrane Phospholipase A (OMPLA) has been suggested to play a role in the virulence of this bacterium. The aim of this study was to profile the most significant cellular pathways and biological processes affected in gastric epithelial cells during 24 h of H. pylori exposure, and to study the inflammatory response to OMPLA+ and OMPLA- H. pylori variants. Results Interleukin-8 was the most significantly up-regulated gene and appears to play a paramount role in the epithelial cell response to H. pylori infection and in the pathological processes leading to gastric disease. MAPK and NF-kappaB cellular pathways were powerfully activated, but did not seem to explain the impressive IL-8 response. There was marked up-regulation of TP53BP2, whose corresponding protein ASPP2 may interact with H. pylori CagA and cause marked p53 suppression of apoptosis. Other regulators of apoptosis also showed abberant regulation. We also identified up-regulation of several oncogenes and down-regulation of tumor suppressor genes as early as during the first 24 h of infection. H. pylori OMPLA phase variation did not seem to influence the inflammatory epithelial cell gene response in this experiment. Conclusion In whole genome analysis of the epithelial response to H. pylori exposure, IL-8 demonstrated the most marked up-regulation, and was involved in many of the most important cellular response processes to the infection. There was dysregulation of apoptosis, tumor suppressor genes and oncogenes as early as in the first 24 h of H. pylori infection, which may represent early signs of gastric tumorigenesis. OMPLA+/-did not affect the acute inflammatory response to H. pylori.

Abstract Background Questionnaires are commonly used to collect patient, or user, experiences with health care encounters; however, their adaption to specific target groups limits comparison between groups. We present the construction of a generic questionnaire (maximum of ten questions) for user evaluation across a range of health care services. Methods Based on previous testing of six group-specific questionnaires, we first constructed a generic questionnaire with 23 items related to user experiences. All questions included a "not applicable" response option, as well as a follow-up question about the item's importance. Nine user groups from one health trust were surveyed. Seven groups received questionnaires by mail and two by personal distribution. Selection of core questions was based on three criteria: applicability (proportion "not applicable"), importance (mean scores on follow-up questions), and comprehensiveness (content coverage, maximum two items per dimension). Results 1324 questionnaires were returned providing subsample sizes ranging from 52 to 323. Ten questions were excluded because the proportion of "not applicable" responses exceeded 20% in at least one user group. The number of remaining items was reduced to ten by applying the two other criteria. The final short questionnaire included items on outcome (2), clinician services (2), user involvement (2), incorrect treatment (1), information (1), organisation (1), and accessibility (1). Conclusion The Generic Short Patient Experiences Questionnaire (GS-PEQ) is a short, generic set of questions on user experiences with specialist health care that covers important topics for a range of groups. It can be used alone or with other instruments in quality assessment or in research. The psychometric properties and the relevance of the GS-PEQ in other health care settings and countries need further evaluation.

Abstract Background International health policy surveys are used to compare and evaluate health system performance, but little is known about the effects of non-response. The objective of this study was to assess the effects of non-response in the Norwegian part of the Commonwealth Fund international health policy survey in 2009. Methods As part of an international health policy survey in 2009 a cross-sectional survey was conducted in Norway among a representative sample of Norwegian general practitioners. 1 400 randomly selected GPs were sent a postal questionnaire including questions about the Norwegian health care system, the quality of the GPs' own practice and the cooperation with specialist health care. The survey included three postal reminders and a telephone follow-up of postal non-respondents. The main outcome measures were increase in response rate for each reminder, the effects of demographic and practice variables on response, the effects of non-response on survey estimates, and the cost-effectiveness of each reminder. Results After three postal reminders and one telephone follow-up, the response rate was 59.1%. Statistically significant differences between respondents and non-respondents were found for three variables; group vs. solo practice (p = 0.01), being a specialist or not (p < 0.001) and municipality centrality (least central vs. most central, p = 0.03). However, demographic and practice variables had little association with five outcome variables and the overall survey estimates changed little with additional reminders. In addition, the cost-effectiveness of the final reminders was poor. Conclusions The response rate in the Norwegian survey was satisfactory, and the effect of non-response was small indicating adequate representativeness. The cost-effectiveness of the final reminders was poor. The Norwegian findings strengthen the international project, but restrictions in generalizability warrant further study in other countries.

Abstract Background X-linked Charcot-Marie Tooth (CMT) is caused by mutations in the connexin32 gene that encodes a polypeptide which is arranged in hexameric array and form gap junctions. Methods We describe two novel mutations in the connexin32 gene in two Norwegian families. Results Family 1 had a c.225delG (R75fsX83) which causes a frameshift and premature stop codon at position 247. This probably results in a shorter non-functional protein structure. Affected individuals had an early age at onset usually in the first decade. The symptoms were more severe in men than women. All had severe muscle weakness in the legs. Several abortions were observed in this family. Family 2 had a c.536 G>A (C179Y) transition which causes a change of the highly conserved cysteine residue, i.e. disruption of at least one of three disulfide bridges. The mean age at onset was in the first decade. Muscle wasting was severe and correlated with muscle weakness in legs. The men and one woman also had symptom from their hands. The neuropathy is demyelinating and the nerve conduction velocities were in the intermediate range (25–49 m/s). Affected individuals had symmetrical clinical findings, while the neurophysiology revealed minor asymmetrical findings in nerve conduction velocity in 6 of 10 affected individuals. Conclusion The two novel mutations in the connexin32 gene are more severe than the majority of previously described mutations possibly due to the severe structural change of the gap junction they encode.

Abstract Background Respiratory syncytial virus (RSV) is a major cause of viral bronchiolitis in infants worldwide, and environmental, viral and host factors are all of importance for disease susceptibility and severity. To study the systemic host response to this disease we used the microarray technology to measure mRNA gene expression levels in whole blood of five male infants hospitalised with acute RSV, subtype B, bronchiolitis versus five one year old male controls exposed to RSV during infancy without bronchiolitis. The gene expression levels were further evaluated in a new experiment using quantitative real-time polymerase chain reaction (QRT-PCR) both in the five infants selected for microarray and in 13 other infants hospitalised with the same disease. Results Among the 30 genes most differentially expressed by microarray nearly 50% were involved in immunological processes. We found the highly upregulated interferon, alpha-inducible protein 27 (IFI27) and the highly downregulated gene Charcot-Leyden crystal protein (CLC) to be the two most differentially expressed genes in the microarray study. When performing QRT-PCR on these genes IFI27 was upregulated in all but one infant, and CLC was downregulated in all 18 infants, and similar to that given by microarray. Conclusion The gene IFI27 is upregulated and the gene CLC is downregulated in whole blood of infants hospitalised with RSV, subtype B, bronchiolitis and is not reported before. More studies are needed to elucidate the specificity of these gene expressions in association with host response to this virus in bronchiolitis of moderate severity.

Background: Rapid technological developments, increased complexity, and increased demand have made patient safety a challenge in radiology.Purpose: To uncover the causes and consequences behind patient injury compensation claims in the use of MRI, CT, and conventional X-ray examinations, and to determine the system factors that need to be focused on in order to prevent these events.Material and Methods: This descriptive cross-sectional study uses data acquired from The Norwegian System of Patient Injury Compensation. A total of 240 cases from 2012-2016 were included.Results: According to our study, the main factors contributing to patient injury compensation claims in radiology were false-negative findings (48.7%), misinterpretation (13.1%), and "satisfaction of search" (12%). Another source of error was routines (8.7%), mainly where the patient should have been (further) examined using another modality. Other causes were related to communication (7.6%), procedures (2.9%), technical factors (2.5%), organizational and management factors (1.5%), competence (0.7%), location of the lesion (0.7%), patient factors (0.7%), false-positive findings (0.4%), and work environment (0.4%). These events led to delayed diagnosis and/or treatment in the range of 0-3650 days.Conclusion: Errors of perception (false negative and "satisfaction of search") and cognitive errors (misinterpretation) were the main reasons behind patient injury compensation claims in radiology. We suggest that a combination of double-reading, specialization, increased collaboration between professionals, as well as a reduction of unnecessary examinations should be considered to reduce adverse events in radiology. [ABSTRACT FROM AUTHOR]