Necator americanus and Schistosoma mansoni in a region of southeastern Brazil.Individuals living in areas endemic for helminths are commonly infected with multiple species. Despite increasing emphasis given to the potential health impacts of polyparasitism, few studies have investigated the relative importance of household and environmental factors on the risk of helminth co-infection. Here, we present an investigation of exposure-related risk factors as sources of heterogeneity in the distribution of co-infection with N. americanus and S. mansoni in the region. Bayesian hierarchical models were then developed to identify risk factors for mono- and co-infection in relation to community-based survey data to assess their roles in explaining observed heterogeneity in mono and co-infection with these two helminth species.Cross-sectional parasitological and socio-economic data from a community-based household survey were combined with remotely sensed environmental data using a geographical information system. Geo-statistical methods were used to explore patterns of mono- and co-infection with N. americanus (71.1%) and/or S. mansoni (50.3%) infection; 41.0% of individuals were co-infected with both helminths. Prevalence of co-infection with these two species varied substantially across the study area, and there was strong evidence of household clustering. Hierarchical multinomial models demonstrated that relative socio-economic status, household crowding, living in the eastern watershed and high Normalized Difference Vegetation Index (NDVI) were significantly associated with N. americanus and S. mansoni co-infection. These risk factors could, however, only account for an estimated 32% of variability between households.The majority of individuals had N. americanus and S. mansoni co-infection between households cannot be entirely explained by exposure-related risk factors, emphasizing the possible role of other household factors in the heterogeneous distribution of helminth co-infection. Untangling the relative contribution of intrinsic host factors from household and environmental determinants therefore remains critical to our understanding of helminth epidemiology.Our results demonstrate that variability in risk of  Necator americanus and Schistosoma mansoni are among the most prevalent of chronic human infections in the developing world. Individuals living in endemic areas are commonly infected with both species. Although the implications of being co-infected with helminths are increasingly recognized, factors influencing patterns of co-infection within human communities remain ill-defined. Here, we describe spatial patterns and risk factors for co-infection with N. americanus and S. mansoni in a co-endemic area in south-eastern Brazil. The prevalence of co-infection with these two helminths in this region was high (41%), varied across the study area and was clustered in high-risk households. We reveal that factors associated with lower socio-economic status  and residential environment (living in the eastern watershed or in areas with less vegetation) were significantly associated with the risk of co-infection relative to being uninfected with either species. Importantly, much of the variability in risk between households (i.e. household clustering) could not be readily explained by these risk factors. The results suggest that, whilst measures aimed at reducing exposure to infection may have an important impact on co-infection and its associated morbidity, untangling the relative contribution of intrinsic host factors (e.g. immune response) from household and environmental determinants remains critical to our understanding of helminth epidemiology.Helminth species such as  Ascaris lumbricoides and Trichuris trichiura were at increased risk of stunting Schistosoma mansoni and two or three soil-transmitted helminth (STH) infections was significantly higher that those harbouring single STH species People living in poor areas of the tropics commonly harbour multiple parasitic infections, including infection with multiple helminth species Recent interest in the scientific study of polyparasitism has given renewed prominence to some old epidemiological questions; in particular identifying factors governing patterns of infection. A wealth of epidemiological investigation across numerous ecological and socio-economic settings indicate that certain characteristics are common to the epidemiology of single helminth species in communities, including household clustering and spatial heterogeneity A. lumbricoides is also endemic to the region, we focus specifically on co-infection with the hookworm Necator americanus and S. mansoni since these species both contribute to iron-deficiency anaemia  and were initially examined using the formalin-ether sedimentation technique for the presence of helminth eggs. Individuals positive for any helminth infection were subsequently examined by the Kato–Katz faecal thick smear technique to quantify the intensity of the infection expressed as eggs per gram of faeces (epg). Two slides were taken from each day's faecal sample for a total of up to four slides from each individual. Morphological examination of expelled worms following treatment among a sub-sample of individuals showed that hookworm infection was exclusively of the species http://edcdaac.usgs.gov/aster/asterdataprod.asp). ASTER provides information on Normalized Difference Vegetation Index (NDVI), a proxy of vegetation density and soil moisture, and digital elevation Household locations were mapped using a hand-held Trimble GeoExplorer global positioning system (GPS) receiver  and ArcPad 6.0.3 software . Readings, with a resolution of 5 m, were taken at the front door, or as near as possible in order to receive a sufficient satellite reception and an average of 10 readings of the co-ordinates were taken. Remotely sensed proxy environmental data were extracted for May 2001 from the Advanced Spaceborne Thermal Emission and Reflection Radiometer (ASTER) satellite sensor at 30 m spatial resolution , and the ethical review boards of George Washington University (USA) and London School of Hygiene and Tropical Medicine (UK). Individuals found to be infected with any soil transmitted helminth or with S. mansoni or N. americanus if at least one egg was detected by either formalin-ether sedimentation or Kato–Katz faecal thick smear. Participants were classified into five age-groups: under 5 years, younger children (5–9 years), older children (10–19 years), adults (20–59 years) and over 60 years. Information on ownership of household assets was used to construct a wealth index using principal component analysis, using the method of Filmer and Pritchett Participants were recorded as positive for an infection with N. americanus, (iii) mono-infection with S. mansoni and (iv) co-infection with N. americanus and S. mansoni. In order to assess the importance of demographic, socio-economic and environmental risk factors on the occurrence of mono- and co-infection simultaneously we used a multinomial modelling approach, which extends logistic regression by estimating the effects of explanatory variables on the probability that the outcome is in a particular category. Initially, for each covariate frequentist unadjusted multinomial models were fit on the outcome in Stata 9.1 , and covariates with P>0.2  were excluded from further analysis. Standard errors were adjusted for dependence between individuals within households. Scatter-plots and the entry of categorised predictor variables were used to investigate non-linear relationships.As an outcome measure a  multi-categorical response for infection status was constructed as follows: (i) no infection, (ii) mono-infection with Subsequently, the retained covariates were built into a Bayesian multinomial mixed effect model in WinBUGS Version 14 . To account for dependence of individuals within households, household was included as a random effect. We employed a Bayesian Monte Carlo Markov Chain (MCMC) approach, which readily allows the development of complex random effects models within households, because, whilst individuals in the same household may have more similar risk than individuals in different households, household-level risk may or may not be spatially autocorrelated. In order to examine the spatial structure of co-infection with N. americanus and S. mansoni at the household level, semi-variograms were generated using the R module GeoR on the basis of household prevalence of mono-infection with N. americanus and S. mansoni and co-infection with both parasites. Before variography, the data was de-trended by regressing against longitude and latitude, in order to remove large-scale spatial trends.Spatial heterogeneity (or structure) refers to the spatially non-random distribution of infection across the study region, such that an individual's risk of infection may be more similar to those living close to them that those living farther away. Such spatial clustering is not necessarily synonymous with clustering Semi-variograms present the semi-variance  of pairs of observations that are separated by the same distance; thus, describing how similar observations are at different spatial distances Of the 1687 residents of the mapped households, 1539 individuals provided stool samples. Sixteen households (59 residents) in the far south-east of the study site were excluded from analysis because cloud-free satellite data were not available. Socio-economic data were unavailable for a further 275 individuals, who mainly lived in the urban municipality. As such, 1208 individuals living in 275 households had complete data. Households with GPS positions less than 10 m apart were treated as a single spatial unit, providing data for 230 locations for spatial analysis, the largest of which had 16 residents.N. americanus, 50.3% had S. mansoni and 41.0% of individuals were co-infected with both helminths (co-infection). 30.1% were infected with only N. americanus, and only 9.4% of individuals were infected with only S. mansoni. The prevalence of co-infection was significantly higher among males than females (p<0.001) and increased significantly with increasing age, peaking among persons aged 20–59 years (p<0.001)  . The occN. americanus or S. mansoni or co-infection with both. The highest frequencies of co-infection were observed in the east of the study area, with an overall prevalence of 86.3% compared to 13.7% in the western watershed. To investigate the global spatial structure of infection patterns semi-variograms were estimated on the basis of household prevalence of mono-infection with N. americanus and S. mansoni and co-infection with both parasites. After removal of the large-scale spatial trend (by regressing against longitude and latitude) there was an apparent lack of any spatial structure for both N. americanus and S. mansoni mono-infection across all separation distances (not shown). Likewise, the semi-variogram for co-infection provides no evidence of spatial dependency, indicating that once the large-scale trends were removed there was no general spatial structure in the distribution of co-infection (nfection .p<0.01).Relative frequencies of household and environmental factors are shown in N. americanus, but not for S. mansoni mono-infection, while residential environment was associated with both mono-infections.Posterior estimates from the adjusted analysis using a hierarchical Bayesian multinomial mixed effects model confirm that the risk of co-infection relative to being uninfected was highest among males and adults aged 20–59 years . There wN. americanus mono-infection (i.e. inclusion of these covariates reduced the household-level variance parameter iu by 40%), substantially less household heterogeneity was explained by these factors for S.mansoni mono-infection (8%) and co-infection (10%). In contrast, environmental factors (living in the eastern watershed and areas with low NDVI) explained 36% of household-level variation in N. americanus mono-infection, 45% in S.mansoni mono-infection but only 19% in co-infection. Household and environmental factors jointly explained 54% of household variation in N. americanus mono-infection, but only 39.5% for S. mansoni mono-infection and 31.9% for co-infection.There was significant household clustering for all outcomes, as indicated by estimates for the household level random effects; the highest degree of unexplained household-level variation was observed for co-infection. Household-level variance was substantially higher when household and environmental risk factors were excluded from the model . Whilst N. americanus and S. mansoni. Our multi-level approach has the advantage of taking into account household clustering of infection, a commonly observed feature of helminth epidemiology We employed a combination of spatial statistics and hierarchical multinomial modelling to investigate spatial patterns and household and environmental factors influencing occurrence of mono- and co-infection by the helminths N. americanus and S. mansoni. While household clustering of single helminth infections is well-documented N. americanus and S. mansoni is more common in households with lower relative socio-economic status is consistent with a study among schoolchildren in rural Cote d'Ivoire, which investigated school-level patterns in co-infection with N. americanus and S. mansoniThe results presented in Our data demonstrated a dominant spatial trend (NE-SW) in household prevalence of co-infection , but theS. mansoni rather than N. americanus, which is more homogeneously distributed across the study area. The high prevalence of S. mansoni in the east of the study region is likely to reflect the increased infectivity of water bodies in this area. It has been frequently demonstrated that within communities high intensity Schistosoma infections can be found clustered around water bodies such as rivers and lakes The dominant NE-SW trend in co-infection observed reflects the distribution of In terms of extrapolation to other settings, Americaninhas is representative of areas of rural northeast Minas Gerias state where helminth infections are highly endemic. Factors which may vary in other settings include contrasting socio-economic and environmental conditions, giving rise to different patterns and risk factors. However, our adopted analytical approach provides a robust methodology to further investigate the epidemiology of polyparasitism in other settings. A final potential limitation of our study, which applies to all multinomial analyses, is the assumption of Independent Irrelevant Alternatives (IIA), which essentially states that the risk associated with each outcome will not change if a new outcome is introduced. However, we believe that this analysis should not be restricted by IIA because our four choices exhaust the available responses (there are no other possible outcomes involving these two infections) N. americanus and S. mansoni was considerably greater than for mono-infection with either N. americanus or S. mansoni. Although this may simply be a reflection of additive household variation associated with each species, it may alternatively be indicative of household factors specifically influencing risk of co-infection. For example, extrinsic factors such as location and infectivity of household water-sources A key finding of our study was that household and environmental risk factors could only account for an estimated 32% of variation between households in the risk of co-infection. Furthermore, unexplained household-level variation of co-infection with N. americanus infection intensity was attributed to genetic factors S. mansoni infection intensity Schistosoma. japonicum, Trichuris trichuria and A. lumbricoides) was in part explained by both genetic  and household (9%) components. This was however a post-treatment study setting, hindering interpretation of results and preventing analysis of infection intensity.Unaccounted household-level variability may alternatively be explained by intrinsic host-related factors such as genetics Household clustering of helminth infection may also be influenced by genetic heterogeneity in the parasite population. Numerous molecular studies have revealed allelic and nucleotide diversity in the genomes of human helminth parasite populations Whilst studies at micro-epidemiological scales are less useful for mapping and prediction of the distribution of co-infection, they are valuable in identifying why certain individuals within communities are at increased risk of multiple helminth infection, and as such have an increased risk of morbidity Appendix S1(0.03 MB PDF)Click here for additional data file.


Crimean-Congo hemorrhagic fever virus (C-CHFV) strains were isolated from a fatal case and the attending physician in Kosovo, Yugoslavia. Early, rapid diagnosis of the disease was achieved by reverse transcription-polymerase chain reaction. The physician was successfully treated with oral ribavirin. These cases yielded the first genetically studied C-CHFV human isolates in the Balkans. Crimean-Congo hemorrhagic fever virus  have been reported in several countries. This widespread virus has been found in Africa, Asia, the Middle East, and eastern Europe. C-CHFV was first identified in the Balkans in 1954. In 1973, Gligic et al. isolated three C-CHFV strains  from two Hyalommaplumbeum plumbeum and one castor bean tick (Ixodes ricinus) Crimean-Congo hemorrhagic fever (C-CHF) is a tickborne hemorrhagic fever with documented person-to-person transmission and a case-fatality rate of approximately 30%. Nosocomial outbreaks caused by gastrointestinal tract, she was transferred to the Institute for Tropical and Infectious Diseases in Belgrade, where she died a few hours after admission.On June 30, 2001, a 43-year-old woman (the patient with the index case), living in Kosovska Mitrovica, Kosovo, became severely ill and was admitted to the hospital with high fever, headache, nausea, generalized myalgia, and disorientation. Two days later her condition rapidly deteriorated; bleeding from the Blood cultures were negative for bacterial pathogens. C-CHF and hemorrhagic fever with renal syndrome (HFRS) were suspected, considering the clinical picture of the patient and the epidemiology of the diseases in this geographic area. A serum sample collected before death was tested for immunoglobulin (Ig) G and IgM specific for a variety of microorganisms  by enzyme-linked immunosorbent assay and indirect immunofluorescent assay (IFA). No specific antibodies were detected.To detect C-CHFV antibodies, the patient’s serum was tested in twofold dilutions  with fluorescein-labeled goat anti-human immunoglobulin  on spot slides containing Vero E6 cells (ATCC CRL 1586), with approximately 50% of the cells infected with the prototype Nigerian C-CHFV strain (IbAr 10200). Titers were recorded as the greatest dilution of serum at which characteristic cytoplasmic immunofluorescence is detected. No specific antibodies were detected.exanthema of soft palate, pneumonia symptoms, leukopenia, and thrombocytopenia 9/L; erythrocyte count: 80x109/L).. We observed a similarity to C-CHFV S segment sequences.We extracted viral RNA from serum and whole blood samples from both patients. Complementary DNA was amplified by nested polymerase chain reaction (PCR) by using primers from the small (S) RNA segment of C-CHFV Since strong evidence exists that ribavirin is effective for treating C-CHF ,6, oral The patient was placed in isolation, and all personnel wore protective clothing, such as gowns, masks, and gloves. All contact persons in the hospital were informed and advised to contact a physician if any symptoms occurred. No additional persons became ill. Samples taken from the driver and the nurse who transferred the woman from the local hospital to Belgrade were tested by IFA for the presence of antibodies to C-CHFV and were found negative, even a month after the woman’s death.Whole blood of both C-CHF cases  was injected separately into 25-mL flasks containing Vero E6 cells. The cells were checked for C-CHFV antigen by immunofluorescence assay with hyperimmune human globulin against C-CHFV (strain Hodja). Positive cells were detected in both flasks on the 6th day after injection. The supernatant of the cells was used to infect fresh E6 cultures, and the virus was passaged six further times. We designated the virus from the fatal C-CHF case as C-CHFV/Kosovo/9553/2001, and the virus from the infected physician as C-CHFV/Kosovo/9717/2001.Extraction of viral RNA of cell culture supernatants, amplification, and phylogenetic analysis showed that the obtained sequences were identical to each other and to the sequences amplified from the clinical samples. The nucleotide sequences were submitted to GenBank and assigned accession numbers AF428144 (for the index case) and AF428145 (for the secondary case). Phylogenetic analysis showed that these sequences clustered together with Drosdov strain (a C-CHFV strain isolated from blood of a patient in Russia), differing from it by 4%, while the genetic difference from the prototype C-CHFV Nigerian strain IbAr 10200 was 17% .Because of the severity of C-CHF, this illness has been difficult to study, either for diagnostic or therapeutic improvements. Antibodies to the virus are not present until 5 days after onset of illness. In addition, patients who have died of C-CHF do not usually develop a measurable antibody response. In these patients, diagnosis is achieved either by isolation of the virus from blood or tissue samples taken in the first days of illness or by molecular methods. In our first patient, who died, no antibodies were detected. Similarly, no antibodies were present during the first days of illness in the secondary case. Immunoprecipitation used for antigen detection gave the first evidence for C-CHF infection in the secondary case. However, reverse transcription (RT)-PCR established the diagnosis of the disease in both cases; sequencing the PCR amplicons confirmed the result. To our knowledge, this secondary case is the first in which PCR was used for rapid diagnosis, although use of PCR on stored acute-phase samples of infected patients has been described ,8.In addition, phylogenetic analysis was informative in comparing the virus strains at the genetic level, giving insights into the molecular epidemiology of the disease. The Greek C-CHFV strain (AP92), not yet associated with disease in humans, differs greatly from the Kosovo strains (24.3% nucleotide difference), forming an independent phylogenetic clade. Isolation of the virus, the standard for diagnosis, was achieved in both cases.The first evidence of the in vitro activity of ribavirin against C-CHFV and its prophylactic use in patients’ contacts occurred in 1989 Although the risk for fatal outcome of the secondary case was not as high, based on criteria described by Swanepoel et al. Concerning the origin of infection of the primary case, no details are available, such as a tick bite or any contact with C-CHF cases in the rest of Kosovo, where a C-CHF outbreak was taking place. No other C-CHF case had been reported from this region. The first patient died a few hours after admittance to the hospital, and the number of contact persons was limited. No one had any symptoms or C-CHF antibodies, except the physician who performed the intubation. He probably came in contact with patient’s excreta, despite protective measures.No tertiary case was detected, as in C-CHF nosocomial outbreaks in other countries  ,11–13. TFurther studies are needed both in diagnostic and treatment fields to elucidate this life-threatening disease. In addition, studies at the molecular level might relate the genetic variation with the pathogenicity of the strains, rendering feasible the production of an effective vaccine.


In the past decades, various protein subcellular-location (SCL) predictors have been developed. Most of these predictors, like TMHMM 2.0, SignalP 3.0, PrediSi and Phobius, aim at the identification of one or a few SCLs, whereas others such as CELLO and Psortb.v.2.0 aim at a broader classification. Although these tools and pipelines can achieve a high precision in the accurate prediction of signal peptides and transmembrane helices, they have a much lower accuracy when other sequence characteristics are concerned. For instance, it proved notoriously difficult to identify the fate of proteins carrying a putative type I signal peptidase (SPIase) cleavage site, as many of those proteins are retained in the cell membrane as N-terminally anchored membrane proteins. Moreover, most of the SCL classifiers are based on the classification of the Swiss-Prot database and consequently inherited the inconsistency of that SCL classification. As accurate and detailed SCL prediction on a genome scale is highly desired by experimental researchers, we decided to construct a new SCL prediction pipeline: LocateP.[LocateP combines many of the existing high-precision SCL identifiers with our own newly developed identifiers for specific SCLs. The LocateP pipeline was designed such that it mimics protein targeting and secretion processes. It distinguishes 7 different SCLs within Gram-positive bacteria: intracellular, multi-transmembrane, N-terminally membrane anchored, C-terminally membrane anchored, lipid-anchored, LPxTG-type cell-wall anchored, and secreted/released proteins. Moreover, it distinguishes pathways for Sec- or Tat-dependent secretion and alternative secretion of bacteriocin-like proteins. The pipeline was tested on data sets extracted from literature, including experimental proteomics studies. The tests showed that LocateP performs as well as, or even slightly better than other SCL predictors for some locations and outperforms current tools especially where the N-terminally anchored and the SPIase-cleaved secreted proteins are concerned. Overall, the accuracy of LocateP was always higher than 90%. LocateP was then used to predict the SCLs of all proteins encoded by completed Gram-positive bacterial genomes. The results are stored in the database LocateP-DB .LocateP is by far the most accurate and detailed protein SCL predictor for Gram-positive bacteria currently available. In bacteria, secreted proteins are involved in stress sensing, substrate binding, cell communication, microbe-host interaction, adhesion, and other essential processes relevant to the environment and life style of the organisms. The secreted proteins are exported via various mechanisms and are retained by the bacterial cell via various interactions or released to the medium Figure . To idenet al. [et al. [et al. [Computational methods have gained considerable precision in the past decades. Initial tools focussed on detecting the presence, type and location of protein transmembrane segments, including signal peptides for targeting and translocation of proteins. One of the very first SCL prediction methods was introduced by Kyte and Doolittle  in 1982 et al.  combined [et al.  combined [et al.  combinedUnfortunately, as a result of the trade-off between specificity and accuracy, computational methods will always be prone to error. Moreover, the number of false predictions increases even further when the SCL-related sequence characteristics have not been properly identified. For instance, among the Sec-dependent exported proteins, current predictors have severe difficulties to distinguish the proteins that are cleaved from the cell membrane by the type I signal peptidase (SPIase) – in this paper we will refer to these proteins as "secreted" – from a relatively large group of membrane-anchored proteins that also contain a putative SPIase-cleavage site but are not cleaved – in this paper we will lump these proteins in the category "N-anchored" -43.As knowledge on the precise SCL of a protein is especially important to judge the biological nature and role of its activity, we constructed a new SCL prediction pipeline called LocateP. Our pipeline is geared to identify the detailed SCL of bacterial proteins by combining existing and novel prediction tools. Special effort was made to increase the accuracy of the prediction of N-anchored proteins. The version of LocateP presented here focuses on SCL prediction of proteins from Gram-positive bacteria.A major drawback of most current sub-cellular location (SCL) predictors is that they are not aimed at the prediction of very specific SCLs but merely at the rather broad locations intracellular, membrane bound/associated and extracellular, in line with the Swiss Prot classification system. We therefore constructed a SCL predictor pipeline LocateP, that distinguishes 7 SCLs and 3 targeting pathways that can be identified in Gram-positive bacteria, with a focus on extracellular SCLs  secretion pathway prediction, 2) transmembrane-segment detection, (3) signal peptide identification, and (4) cleavage and retention signal recognition. The LocateP pipeline employs existing SCL prediction tools . The Weblogos [To identify the features that determine cleavage, the multiple sequence alignments of the signal peptides from experimentally validated N-anchored and secreted proteins  containiWeblogos  of the tet al. [Ideally, the performance of the LocateP pipeline should be checked with large experimentally validated data sets. Unfortunately, the availability of such large data sets is rather limited. LocateP was tested first with the experimental data set of Tjalsma et al.  which waBacillus subtilis 168, Bacillus cereus ATCC14579 and Lactobacillus plantarum WCFS1 was verified . LocateP clearly performed better than the other two specific tools when tested with intracellular and membrane proteins sets, and thus showed an excellent capability of Tat-signal peptide detection  to > 90LocateP was designed as a pipeline, and hence could have performed less well on specific categories than specialized tools. In particular, the performance would have been considerably lower if the flow scheme had been chosen wrongly. However, a comparison of the performance on lipoproteins, membrane proteins, Sec-secreted and Tat-secreted proteins with the specialized tools LipoP 1.0, TMHMM 2.0, Phobius, SignalP 3.0, Predisi, Tat-find v.1.2 and TatP shows that LocateP does not suffer from being a pipeline tool. Apparently, our choice to mimic the order in the bacterial secretion process was a correct one. In fact, it has been shown by others that the SCL prediction can be improved considerably by simulating the protein sorting processes ,102. OveBecause of the high prediction accuracy of LocateP on proteins of known biological function . LocateP showed slightly lower accuracy than SigTree did, which suggests that the sequence composition of signal peptides from Gram-positive and Gram-negative bacterial proteins could be different. Future versions of LocateP will be improved and extended to Gram-negative bacterial protein SCL prediction by incorporating Gram-negative specific subcellular-location prediction tools.LocateP was first tailored for the SCL prediction of Gram-positive bacterial proteins; therefore prediction of the Gram-negative specific proteins, such as β-barrel membrane proteins, was not yet included in the pipeline. LocateP was compared to SigTree , a signaFinally, we must emphasize that in several cases an automatic SCL prediction of a protein will inevitably give an incorrect prediction using LocateP or any other tool: (i) when the start codon of a gene encoding a protein with a signal peptide has been wrongly identified (either too far upstream or too far downstream), (ii) when a frame shift in the open-reading frame leads to different fragments of encoded proteins, and (iii) when an intracellular protein contains a signal peptide-like hydrophobic helix near the N-terminus; in this case such helices generally fold into the interior of the globular protein -144.As detailed and accurate genome-scale SCL prediction of encoded proteins is highly desired by scientists in various biological research areas, numerous existing and newly developed tools were combined into one pipeline: LocateP. To date, LocateP is the most detailed protein SCL predictor for Gram-positive bacterial proteins among all tools that have been reported, in that it presently distinguishes 7 different SCLs and 3 sorting pathways, with focus on extracellular SCLs. Moreover, it is also the most accurate SCL predictor, especially on distinguishing N-anchored and secreted proteins. LocateP was applied on all completed Gram-positive bacterial genomes from the NCBI sequence database. The results are updated synchronously with Genbank updates and are publicly available via the database LocateP-DB . The preth, 2007 ("ORGANISM" annotation fields: 'Firmicutes' or 'Actinobacteria'), and were continuously updated since then. Protein sequences of Bacillus species were collected both from GenBank on April 1st, 2007 and from the ERGO database [th, 2006.The genome sequences of Gram-positive bacteria were extracted from GenBank on May 30database  on Novemth, 2007, and an expert evaluation against the protein function annotation as retrieved from GenBank on August, 10th, 2007.Eight different protein data sets of known subcellular location were selected from literature describing other tools and describing proteome studies Table  legend. Multiple sequence alignments were built with MUSCLE . HMMs weMany studies have compared and evaluated currently available transmembrane segment and signal peptide predictors -100,148.LocateP detects signal peptides by scanning the protein N-terminus, which was defined as the initial 60 amino acids of the protein, using SignalP 3.0, Phobius and PrediSi. Some proteins have a signal peptide shortly after these 60 amino acids. These proteins were predicted as "intracellular", but we added the extra remark of "TMH start AFTER 60" to the annotation indicating that these proteins could be secreted. No attempt was made to choose alternative start codons of incorrectly predicted start sites of ORFs.et al. have experimentally determined the SCL of a large number of Bacillus subtilis proteins [Recently, Tjalsma proteins . The expproteins  in the Eproteins  using fuPairs of HMMs were built to separate the group of proteins with a putative SPI cleavage-site into those that are cleaved (i.e. secreted/released) and those that are not (i.e. N-anchored). The sequences were aligned around the putative cleavage site and the length of the HMMs was varied (length >8). All HMM pairs were applied to both "EXP-" sets; the E-value was set at 10,000 to assure each protein gained an HMM score. For each pair the separation between truly cleaved and truly N-anchored proteins was analyzed and it appeared that the HMM pair containing equivalent amounts of H and C region residues achieved the highest specificity in distinguishing the two sets  each displayed a relatively high specificity, but this was increased significantly by combining the two HMMs. A generic scoring scheme was derived via the following procedure: i) The HMM scores were rounded to discrete integers and the score distribution for the EXP-anchored and EXP-secreted protein sets was used to determine a first cut-off. The discrete HMM scores related to the HMMnon-cleaved ranged from -19 to +20 with all non-cleaved (i.e. N-anchored) proteins scoring higher then 3, those related to the HMMcleaved ranged from -29 to +20 with all cleaved (i.e. secreted) proteins scoring higher then 0. In fact, for both HMMs only in a small scoring range the two protein groups overlapped. Therefore, all sequences with a score ≤2 using the HMMnon-cleaved were attributed the SCL: SEC-secreted, and those with a score ≤-1 using the HMMcleaved were attributed the SCL: N-anchored. ii) For those sequences that scored >2 with the HMMnon-cleaved and >-1 with the HMMcleaved, the score with both models was compared. In case HMMcleaved > HMMnon-leaved score, the sequence was considered SEC-secreted, whereas, in case HMMnon-cleaved ≥ HMMcleaved score, the sequence was considered N-anchored.The individual HMMs of the selected pair  showed the highest specificity when the T-score was set to 3. Gaps were allowed in this model except in the region of the lipobox (residue -5 to the lipobox Cysteine)The experimental data of Tjalsma et al.  indicateparanoid  search fparanoid ,157,158,et al. [E. coli [Bacillus subtilis [The 105 putative Tat-secreted proteins (according to Swiss-Prot) from the TatP-positive training set  were taket al. , the -3 [E. coli -162, whisubtilis ,164). Thsubtilis ,163,164.The following topological criteria were used to identify LPxTG-type cell-wall anchored and C-anchored membrane proteins. For the selection of LPxTG-anchored proteins, the criteria were : (i) theThe criteria used for predicting C-anchored proteins were: (i) the protein has only 2 predicted TM helices, one situated at the N-terminus and one at the C-terminus, (ii) the protein has a cleaved N-terminal signal peptide, (iii) the protein has a C-terminal transmembrane helix and a positively charged C-terminal tail but no LPxTG motif, (iv) the distance between the N-terminal and C-terminal helices is larger than 45 residues.RS conceived the project and acquired the funding, MM performed the research and constructed the LocateP pipeline and database, JB helped in tool selection, CF and RS supervised the project, while MM, CF and RS wrote the manuscript. All authors read and approved the manuscript.Flow chart and decision tree of the LocateP pipeline. The different SCL tools used at each decision step are indicated. The different SCLs distinguished are boxed in the middle; "Bacteriocin" signifies bacteriocin-like proteins secreted by non-classical pathways, identified by Bagel. a) all tools agreed, b) all possible hits, c) majority vote, d) 2–3 TM segments and C-terminus detected by LPxTG HMM.Click here for fileBacillus subtilis.The LocateP predicted N-anchored and secreted proteins with known function in Click here for fileBacillus subtilis.The LocateP predicted N-anchored and secreted proteins of unknown function in Click here for fileLiterature references for other experimental evidence (Yes (O)) listed in additional file Click here for file


The long-term course of dizziness was investigated combining medical chart and survey data. The survey was undertaken median (interquartile range (IQR)) 4.6 (4.3) years after the initial medical examination.Chart data comprised sex, age, diagnosis, symptom duration, postural sway and neck pain. Survey data comprised symptom severity assessed by the Vertigo Symptom Scale – Short Form (VSS-SF), and data regarding current state of dizziness, medication, neck pain and other chronic conditions.The sample consisted of 503 patients, the mean (standard deviation (SD)) age was 50.0 (11.6) years, women being slightly overrepresented (60%). Severe problems with dizziness (VSS-SF mean (SD) 13.9, (10.8)) were indicated in the total group and in 5 of 6 diagnostic sub-groups. Vertigo/balance- and autonomic/anxiety-related symptoms were present in all groups. Current dizziness was confirmed by 73% who had significantly more severe problems than the non-dizzy (VSS-SF mean (SD): 17.2 (10.1) versus 5.0 (7.3)). Symptoms were related to vertigo/balance more than to autonomic/anxiety (test of interaction p < 0.001).Based on simple logistic regression analysis, sex, symptom duration, neck pain, sway and diagnoses predicted dizziness. Symptom duration and neck pain remained predictors in the adjusted analysis. Age, symptom duration, neck pain, sway and diagnoses predicted vertigo/balance-related dizziness in both regression analyses. Sex, neck pain and sway predicted development of autonomic/anxiety-related dizziness according to simple regression analysis, while only neck pain remained a significant predictor in the adjusted analysis. With respect to diagnosis, simple regression analysis showed significant reduced likelihood for development of dizziness in all vestibular sub-groups when compared to the non-otogenic dizziness group. With respect to vertigo/balance- and autonomic/anxiety-related symptoms, the implication of diagnostic belonging varied. No effect of diagnoses was seen in adjusted analyses.The majority of patients had persistent and severe problems with dizziness. The wait-and-see attitude before referral to specialist institutions may be questioned. Early, active movements seem necessary, and attention should be paid to the presence of neck pain. Diagnoses had limited prognostic value. Questionnaire-based evaluations could assist in classification and identification of type of dizziness and thereby provide a better basis for specific rehabilitation. For many patients experiencing vertigo and dizziness, symptoms resolve spontaneously within a short period of time . AccordiReferral rates from primary care to specialist units vary. The number of visits, length of history  as well The purpose of the present study was to explore the long-term course of dizziness in some patients referred to a balance clinic in a university hospital with complaints of persistent dizziness. It was also an aim to explore the predictive ability of some factors identified in the medical examination.Patients between 18 and 70 years with the following diagnoses in the vestibular category were included in the present study: Menière's disease, vestibular schwannomas, benign paroxysmal positional vertigo and vestibular neuritis. Patients with central vestibular disorders were excluded. Patients with non-vestibular dizziness were assigned to two groups. Those with significant neck symptoms accompanying dizziness were diagnosed as "cervical dizziness" when no other explanation was found. The remaining patients diagnosed in the non-vestibular group were classified as "non-otogenic" when no other cause was found. The inclusion criteria were met by 821 patients.All included patients had been examined for suspected vestibular disorders. Patients were referred from general practice and other specialist care units. The extent of the medical examination varied accordingly, but most of the referred patients had undergone evaluation by an otolaryngologist. The evaluation in our clinic included audiometry (pure tone and speech) and clinical ear, nose and throat examination. The clinical examination was associated with laboratory investigations comprising static posturography, electronystagmography with bithermal caloric tests, testing for spontaneous and positional nystagmus, ocular smooth pursuit and saccades. The final diagnosis was set retrospectively by an experienced otolaryngologist according to medical chart information.The study combined existing medical chart data (1992–2001), and postal survey data collected in spring 2002. The median (interquartile range (IQR)) time period from the patients' first medical examination to the survey, was 4.6 (4.3) years.©, Bologna Italy) [Medical chart data were sex, age, neck pain (yes/no) and balance in the standing position  by static stabilometry  . SymptomSurvey data were age, use of vestibular sedatives, presence of neck pain and of other chronic conditions. Apart from age, the questions were coded as yes/no. Current state of dizziness was identified by the following question: "Are you still bothered with dizziness?" with response categories: Yes/No. Recent spells of dizziness were identified by "When did you have your last episode of dizziness?" with response categories: 1) During the day, 2) Within last week, 3) Within last month, 4) More than a month ago. Categories 1–3 were collapsed to indicate "recent episode", i.e. within the last month. Symptom duration from debut to survey, and time interval between medical examination and survey, were calculated.Severity of symptoms was assessed in the survey by a Norwegian version of the Vertigo Symptom Scale – Short Form (VSS-SF)  developeThe study was approved by the Regional Committee for Medical Research Ethics in Western Norway as part of a larger study. Written informed consent was obtained from all patients.Demographic and survey data were compared between the diagnostic groups, and reported as mean, median, standard deviation (SD), interquartile range (IQR) and 95% confidence interval (CI). Distribution of symptom scores was examined by q-q plots and by comparing mean and median scores of the VSS-SF and sub-scales. As normality could be assumed, parametric statistical methodology was used. Mean sub-scale score was obtained by dividing the sub-scale sum score with the corresponding number of items. One-way analysis of variance (ANOVA) with Bonferroni's post-hoc tests was used to explore differences between groups in normal continuous data, for non-normal continuous data the Kruskal-Wallis test was used. Pearson's chi-square statistical methodology was used to explore differences between groups in dichotomous data. Statistical significance was set at p ≤ 0.05. SPSS version 15 for Windows was used for all analyses.One-way ANOVA with Bonferroni's post-hoc test was performed to determine the significant differences between diagnostic group means. In order to analyse possible dependency of the VSS-SF on the state of dizziness and diagnosis, a two-way ANOVA was performed with dizzy/not dizzy (2 categories) and diagnostic groups (6 categories) as grouping factors. This allowed for testing if symptom score differed in dizzy and not dizzy, and if the difference varied across diagnostic groups (interaction). To analyse the possible dependency of the VSS sub-scales on the state of dizziness and diagnosis, ANOVA was performed with dizzy/not dizzy (2 categories) and diagnostic group (6 categories) as grouping factors, and sub-scale as repeated factor . This allowed for testing complex interactions, e.g. if difference in sub-scale scores between dizzy and not dizzy was the same for both sub-scales, and if it was modified by diagnostic group. The general linear model (GLM) procedure with the repeated measures option of SPSS was applied .and type of dizziness. Dependent variables were VSS-SF, VSS-V and VSS-A using the respective cut-off values indicating the absence or presence of symptoms. Sex, age , neck pain, path length (converted to meters), symptom duration at medical examination  and diagnosis  retrieved from the medical charts were used as explanatory variables. The complete model is presented in the results.Simple (unadjusted) and multiple (adjusted) logistic regression analyses were used to identify predictors of overall dizziness Of 2067 patients, 821 (40%) met the inclusion criteria and 549 (67%) patients returned the questionnaire. Closer inspection showed a varying number of missing items in 86 of the returned VSS-SF forms. Imputation was done in 40 forms, i.e. forms lacking one or two items on the respective sub-scales, 46 forms were discarded. The final sample therefore consisted of 503 responders with sufficient data for calculation of symptom scores on the VSS-SF. In the group, 385 patients responded to the first mailing (early responders), while 118 patients responded to posted reminders (late responders). The remaining 272 patients did not return the questionnaire (non-responders).At the time of the medical examination the median (IQR) symptom duration was 1.8 (4.0) years. The median (IQR) path length measured by the balance platform was 829 (668) mm. Neck pain was reported by 27%. Details of demographic information from medical charts according to diagnostic groups are presented in Table At the time of the survey, the mean (SD) age was 50.0 (11.6) years. Patients with vestibular schwannoma were significantly older than patients in the non-otogenic and cervicogenic groups (p < 0.001). Women were slightly overrepresented (60%), but no difference in sex distribution was seen between the diagnostic groups p = 0.33). Only 8% used vestibular sedatives, 59% n = 298) indicated neck pain, and 61% (n = 299) reported other chronic conditions. The median symptom duration (IQR) was 7.1 (5.5) years. In the total sample, the mean (SD) symptom score on the VSS-SF was 13.9 (10.8); on the VSS-V it was 7.5 (6.8), and on the VSS-A it was 6.4 (5.3). Significant higher scores were seen in the non-otogenic compared to the vestibular schwannoma group on the total and sub-scale scores. Details of scores in the diagnostic groups are presented in Table 98 indica3. Only 8The results from the two- and three-way ANOVA showed that diagnosis could be eliminated as it had no significant effect on the severity of symptoms. As expected, the mean symptom score on the VSS-SF differed significantly (p < 0.01) between dizzy and not dizzy patients; the mean difference was estimated as a score of 12.18 . On the sub-scales, the mean difference in symptom scores between dizzy and not dizzy was larger for the VSS-V than for the VSS-A (test of interaction p < 0.001) estimated as 0.98  on the VSS-V, and as 0.63  on the VSS-A sub-scale. This indicates that vertigo/balance was a greater problem than autonomic/anxiety-related symptoms. Details of scores according to dizziness and diagnostic groups are presented in Table Simple logistic regression analyses showed that sex, symptom duration, neck pain, sway, and diagnosis, but not age, significantly predicted dizziness on the VSS-SF. Moreover, all the variables significantly predicted vertigo/balance-related symptoms, while sex, neck pain and sway predicted autonomic/anxiety-related symptoms. Diagnosis predicted significant reduced likelihood for development of symptoms when comparing non-otogenic dizziness to diagnoses in the vestibular category. The effect of diagnostic belonging with respect to vertigo/balance- and autonomic/anxiety-related symptoms varied. Details of the results are presented in Table The adjusted regression models left long-term symptom duration (> 6 months) and neck pain as significant predictors of dizziness. These two variables, in addition to sway and age, significantly predicted vertigo/balance-related symptoms, with age as a negative predictor. Neck pain was the only significant predictor of autonomic/anxiety-related symptoms Table . DiagnosThere were no significant differences between responders and non-responders (n = 272) with respect to information of age, sex, sway parameters and symptom duration from debut to survey.At the time of the survey, the whole sample had severe symptoms of dizziness years after onset, and dizziness was associated with vertigo/balance- as well as autonomic/anxiety-related symptoms in all diagnostic groups. Significantly more severe dizziness was seen in the non-otogenic compared to the vestibular schwannoma group. The majority (73%) confirmed current dizziness with recent episodes, and the mean (SD) symptom score in this group was 17.2 (10.1) on the VSS-SF scale, which defines severe dizziness ≥ 12 . In the or positive (lower) symptom scores [The severe and persisting dizziness was somewhat surprising, and the results should be interpreted with caution, as one third failed to return the questionnaire. It is possible that an attrition bias may have influenced the results negatively. It has been suggested that non-responders do not bother to reply because of fewer problems . Respondm scores . It is dm scores .Symptom severity in the present study corresponded to findings in another study  with simIn the present study vertigo/balance-related symptoms were a greater problem than autonomic/anxiety-related symptoms in line with findings in the other study from our department . On the The majority of patients had vestibular disorders. The conditions are considered to be benign, and most, with some exceptions, have a short and self-limiting course ,23. In MOne explanation of the severity of symptoms could be related to patient selection, since a large proportion of patients were referred due to persistent symptoms, leading to more secondary health problems and possibly worse prognosis. Symptom severity might also be explained by the presence of co-morbid conditions at the time of the survey, although no association between dizziness and other chronic conditions were found. Physical inactivity over the years could also explain symptom severity. Early exercise as a means to enhance compensation has been documented , and theLong-term symptom duration and neck pain were the most important predictors of symptom development. Short-term duration of symptoms at referral < 6 months) combined with programs of exercises is suggested to influence the prognosis positively . A posit months cAs for neck pain, head instability associated with vestibular disorders  may resuOur study showed that increased sway, i.e. physical signs of balance problems, had some predictive effect on vertigo/balance-related symptoms. Balance is negatively influenced by age , but in Being classified in the non-otogenic category increased the likelihood of developing problems compared to patients in the vestibular category. The first category rules out specific causes of dizziness, which in itself could result in sustained problems for psychological as well as physical reasons; i.e. fear of moving to avoid provocation of dizziness. However, in the adjusted analyses, there was no effect of diagnosis, which is in conformity with two other studies ,36. The Classification of patients with long-lasting dizziness is not straight forward, and strict criteria could not always be applied, due to missing information in patient history data  and unclIn conclusion, this study shows that a large group of patients have persistent and severe dizziness influenced by vertigo/balance- and autonomic/anxiety-related symptoms years after the initial diagnosis. The wait-and-see attitude is problematic, as early and active exercises paying attention to the presence of neck pain is essential to promote compensation. Diagnoses have limited prognostic value. It is suggested that questionnaire-based evaluations could assist in classification of patients and identification of symptoms, and thereby provide a better basis for specific rehabilitation.VSS: Vertigo symptom scale; VSS-SF: Vertigo Symptom Scale – Short Form; VSS-V: vertigo-balance sub-scale; VSS-A: autonomic-anxiety sub-scale; IQR: interquartile range; SD: standard deviation.The authors declare that they have no competing interests.KW designed the studies, collected data, performed statistical analysis and drafted the manuscript. AEL, FG, and SHGN all participated in designing the study and contributed in drafting the manuscript. GEE has participated in the statistical analysis and in drafting the manuscript pertaining to statistics and results. All authors have read and approved the final manuscript.The pre-publication history for this paper can be accessed here:


Past studies have linked certain values  with adolescent substance-use. The aims of this study are to replicate cross-sectional research linking values and adolescent substance-use and to determine if such values predict future substance-use.A longitudinal school-based survey of 2196 young people (age 15) followed up in early adulthood (age 18/19). Participants provided data about their beliefs and values at age 15, and their substance-use  at ages 15 and 18/19. In addition data were collected about their social background .Cross-sectionally, young people with anti-authority values were more likely to use various substances, e.g. 17-67% more likely to regularly smoke (daily), drink (most days), or use drugs (weekly) for each SD above typical levels. Adjusting for social background, associations were not substantially attenuated. However in the prospective analysis, adjusting for both background and substance-use at age 15, only two (anti-authoritarian and work ethic) values were  associated with substance-use at age 18/19.While we replicated results found in prior cross-sectional studies, evidence from this study does not support the argument that holding certain 'pro-social' or 'good' values substantively protects against later substance-use and challenges the likely effectiveness of values-based interventions in relation to later substance-use. Character or values education is the policy of using the school curriculum to influence young people's values, typically promoting traditional and citizenship values. It is a topic which provokes controversy in its own right -3, even Before reviewing the limited evidence linking values and substance-use, we briefly outline the controversies surrounding values or character education, define what 'personal or human values' are and how such values are measured.Combating Violence With Values: The Character Education Solution' [http://news.bbc.co.uk/1/hi/world/europe/8437460.stm).Proponents of values education make strong claims for its effectiveness, as for example in Lickona's 'olution' . These colution' . Proponeolution' . Represeolution'   is covered in the Personal and Social Education curriculum [Much of the debate originates in America, with ex-president George W. Bush among the many advocates of character education . During rriculum .Values education can be described, according to the literature review, as a subject about which much has been written but little is known." (page 33) [The debate is largely conducted in the absence of any reliable evidence, with a recent Australian government review summarising the evidence base thus: "page 33) . Against"desirable goals, varying in importance, that serve as guiding principles in people's lives." [restraint of actions, inclinations, and impulses likely to upset or harm others and violate social expectations or norms" [If there is a lack of empirical evidence in relation to values education, the opposite is true for the psychological study of values. There is extensive cross-cultural agreement regarding the measurement and structure of values, which are broadly defined as  lives." . The add lives." ,9. Briefr norms"  and is mpursuit and protection of the common good) and Excellence (seeking to accomplish something noteworthy and admirable). These are compatible with Schwartz's universalism (humanitarian) and achievement (self-enhancement) values , including some that could be considered health-risks. Achievement values were associated with stress-related behaviours (taking on too many commitments) and hedonism with over-eating. Both hedonism/stimulation and traditional values were strongly linked to aggregate measures of appropriate hedonistic or traditional behaviours (r >.62). Further, when self-reported behaviours were substituted with partner or peer ratings of participants' behaviours the association remained strong, providing further validation. While Bard and Schwartz [Among the few existing studies, there is evidence that specific values relate to a number of everyday  behaviours in a plausible manner. In a convenience sample of undergraduates, Bard & Schwartz  found stSchwartz  provide Even fewer studies have examined the link between young people's values and health-related behaviours, but of these, most concentrate on substance-use. We briefly review this literature, finding strong cross-sectional associations, but also considerable methodological limitations.One difficulty is that much research has conflated different types of risk-behaviours, sometimes creating a composite risk-score from a disparate selection of problem behaviours encompassing sexual behaviour, violence, substance-use, etc . In a reet al. [Using a composite indicator of substance-use in schoolchildren (ages 10-11) Cole et al.  found onet al. . This laet al. ,16, in aIn a study of high school student's (ages 15-16) problem behaviours, achievement (being well-respected) values were associated with lower, and hedonistic (fun and enjoyment) values with higher use of three substances  . Additior = 0.21-.29) and self-enhancement values (r = 0.36) values, but negatively correlated with humanistic  values [One diary study investigating alcohol-use among college students found alcohol consumption moderately correlated with both hedonistic  values . AdditioFew studies have focused exclusively on smoking and values. In a random sample of UK adults, heavy-smokers held stronger achievement and hedonistic  values while light-smokers held more traditional and humanistic  values . A repliMore recently, a study by Chen  examinedIn summary, although the evidence-base is limited (often restricted to student populations), a consistent finding is that traditional values are protective, while self-direction (independence) values are risk factors for substance-use. Self-enhancement (ambition) values can be protective or not dependent upon the context, age group or substance, e.g. they are potentially a risk factor in environments with a work-hard, play-hard ethos such as college. Humanitarian values are also inconsistently associated with substance-use, possibly because of the association between cannabis-use and peace-activist/humanitarian movements.However, the evidence-base remains inadequate for at least two reasons. Firstly, only a few ,21 studiUsing a large representative longitudinal sample of young people, surveyed at ages fifteen and again at 18/19, we have three aims; first, to replicate previous cross-sectional studies finding associations between values and smoking, alcohol-use and drug-use; second, to determine if these associations remain after adjustment for relevant confounds; third, to establish if values predict future substance-use at age 18/19, after adjusting for both confounds and past health-behaviour.The material for the study is drawn from a Scottish longitudinal community health and lifestyle survey of young people, administered first in-school via questionnaire  and then in the post-education period by nurse interview at age 18/19. The focus here is on data collected between 1999 (aged 15) and 2003 (age 18/19) within the framework of the 'West of Scotland 11 to 16 Study/16+' . The stuDue to the school-based nature of the sample the sampling scheme involved several elements to ensure a representative sample at both the primary and secondary school stages and sufficient school units to investigate or control for school-level clustering . BrieflyAt age 11 the sample was representative  of 11 year olds in the study area . DiffereSome equality in marriage is a good thing but by and large the husband ought to have the main say'; Work Ethic (Schwartz's Conformity: self-discipline), e.g. 'Even if I didn't like the work, I would still want to do it as well as I could'; Equity , e.g. 'The government should tax the rich more in order to help the poor'; Citizenship and sense of belonging (Schwartz's Security: sense of belonging), e.g. 'It is a privilege to be Scottish'; Anti-authority (Schwartz's Anti-tradition: no respect for tradition/obedience), e.g. 'Young people today don't have enough respect for traditional values' (reverse scored); Anti-traditional - apolitical or environmentalist - politics , e.g. "There should be restrictions on car drivers in the city to cut down on pollution'; Materialism , e.g. 'There's nothing wrong with having a big house or an expensive car'; Individualism , e.g. 'The idea that society owes you a living is out of date'. The vast majority of these items have been validated in past studies, but they do not originate from established values scales. However, given the universal nature of values and the high face validity of many items it is highly likely our items are strongly correlated with equivalent items drawn from an established values scale e.g. "Parents can tell you what to do" (our item) Vs "Honoring parents"  pupils were asked to rate on a 5-point Likert scale (strongly agree to strongly disagree) how much they endorsed 32 questions relating to values and social attitudes, derived from a number of well-established studies of young people's values -28  to 7 (most deprived), was derived from pupils' postal codes using the 'Carstairs'  index, aWith respect to measures of substance-use at ages 15 and 18/19, smoking was defined as regular smoking, derived from a 5-point frequency scale  and is similar to that used by the UK Office for National Statistics . AlcoholThe analysis used logistic regression to determine the association between values at age 15 and both concurrent and later (age 18/19) substance-use. Analysis was conducted first unadjusted and then adjusted for background factors and included past substance-use for age 18/19 outcomes. We constructed weights to compensate for differential attrition (21), but use of these weights did not alter results, nor did adjusting for school clustering (either via multilevel modeling or adjusting estimates for clustering). The influence of missing data was further explored by comparing results for models using three different methods for dealing with missing data; complete data only; including an additional missing data category for variables with more than modest amounts (50+ cases) of missing data and multiple imputation methods. Multiple imputation was implemented using the STATA 'ice' procedure and included all variables from the relevant model. Categorical variables were imputed using logit or multiple logit, continuous variables using regression and deprivation using ordinal logit commands. Ten imputed datasets were used to calculate the final combined estimates. Although the results for each method were not substantively different, we report results based on multiple imputation. Results from all other alternative models are available upon request.Table Table With the single exception of alcohol-use, both anti-authority and anti-traditional politics predicted increased substance-use of any type, with anti-authority typically the strongest predictor. Traditional sex-roles also predicted increased substance-use irrespective of type, although only in the adjusted model in relation to smoking. Work ethic was associated with increased regular smoking, individualism with lower smoking and drug-use and equity with a significant, but small, increase in smoking.Table Returning to our three aims, we replicated the well-established cross-sectional links between values and substance-use found in the literature. Secondly, we established that background factors did not explain these cross-sectional associations. Finally, but most importantly, although some values do predict future substance-use in simple (unadjusted) analyses, after adjusting for background and prior substance-use almost all the associations either vanish or are reduced to marginal effect sizes.With respect to the cross-sectional findings, the associations between specific values and substance-use in this study are compatible with past research, but intriguingly not all values typically thought to reduce substance-use did so. This raises questions about what values are 'good' in relation to substance-use. As expected, independence and rebellious orientated  values were associated with greater substance use. However, several unexpected associations are more notable, but require explanation. The links between traditional sex-roles and increased substance-use and between work ethic and smoking, suggests that certain cultures, competitive environments, or 'masculine' sex-roles  encouragOur longitudinal findings directly challenge the proposition that authoritarian or traditional values are necessarily better for health than liberal or individualist ideals -3. HowevFrom a developmental perspective it is of course possible that values in childhood or early adolescence may predict uptake before age 15, but given the consensus that values are not fully formed until mid to late adolescence -37 this Should policymakers encourage certain moral values on the assumption that this will inhibit later (early adult) substance-use? The evidence from our study suggests not, or at least not to any significant degree. If the aim is to promote values deemed socially desirable, then this is a wider moral question largely detached from public-health policy. Advocates of values education will likely focus on the cross-sectional results and the two  significant longitudinal associations between anti-authority/political values and substance-use, while their opponents will likely concentrate on the small effect size(s) and the, generally, negative longitudinal findings. We do not wish to discourage innovative approaches to reducing population levels of substance-use among young people, but it is clear from our results that future values-based studies and interventions should be rigorous  and focused on 'practical' significance.Arguably, most people intuitively consider certain values  to be of greater ethical and social 'worth' and these values are to be encouraged. However, expanding this to encompass improved health and health-related behaviours is problematic. Our evidence suggests that these values are not necessarily 'good' (or 'bad') for you, at least in relation to substance-use.All authors are supported financially by the Medical Research Council of Great Britain as part of the Youth and Health Programme (WBS U.1300.00.007) and have no other competing interests.RY wrote the majority of the manuscript, conceived the theoretical approach taken in the paper, participated in design of the final phase of the 11-16/16+ study, and performed the statistical analysis. PW conceived of the original 11-16 study, and participated in its design and coordination and helped to draft the manuscript. Both authors read and approved the final manuscript.The pre-publication history for this paper can be accessed here:http://www.biomedcentral.com/1471-2458/10/165/prepubSupplementary supporting material. Contains 2 figures; figure 1a locates specific items within the Schwartz value model; figure 1b provides a simplified model of Schwartz's circumplex model. Box 1 contains a broad introduction to the Schwartz model and a summary of its 10 values and typical items used in their measurement. Table 1 shows the factor loadings of the 32 items after varimax factor analysis in relation to an 8 factor solution.Click here for fileQuestionnaire items. List of 32 items on opinions and beliefs used in the factor analysis.Click here for file


Fibroblast activation protein (FAP)/seprase and dipeptidylpeptidase-IV (DPP-IV)/CD26 are serine integral membrane proteases. They are involved in tissue remodelling, cancer invasion and metastases, mechanisms that are controversial. The aim was to identify cell types that express FAP and DPP-IV in human bone and soft tissue tumours, and to determine whether there are any correlations between the expression of FAP and DPP-IV and the malignant potential of tumours.in situ expression in 25 malignant and 13 benign human bone and soft tissue tumours. Reverse transcriptase-polymerase chain reaction analyses confirmed mRNA expression of FAP and DPP-IV in all individuals. Immunohistochemistry using pre-fixed frozen sections revealed that FAP was positive in low-grade myofibroblastic sarcoma, the fibroblastic component of osteosarcomas, and malignant fibrous histiocytomas, but negative in Ewing’s sarcomas and rhabdomyosarcomas. DPP-IV showed similar immunohistochemical results. Among benign tumours, non-ossifying fibromas, desmoid tumours and chondroblastomas expressed both FAP and DPP-IV. Giant cells expressed DPP-IV in giant cell tumours.This study analysed Our data suggest that FAP and DPP-IV are consistently expressed in bone and soft tissue tumour cells that are histogenetically related to activated fibroblasts and/or myofibroblasts, irrespective of their malignancy. DPP-IV is also expressed in monocyte–macrophage lineage cells. Fibroblast activation protein (FAP)/seprase is a serine integral membrane proteinase. The molecular weight of FAP is either 95 or 97 kDa as a monomer and 170 kDa as a dimer. The dimer shows gelatinolytic activity.The present study was designed to identify cell types that express FAP and DPP-IV in human bone and soft tissue tumours, and to determine whether there are any correlations between the expression of FAP and DPP-IV and the malignant potential of tumours.Forty-one bone and soft tissue tumours were analysed. All specimens were obtained by open biopsy or surgical resection from patients, none of whom had received radiation or chemotherapy. All tumours were diagnosed on the basis of clinical features, imaging, conventional histopathology and immunohistochemistry of paraffin-embedded sections. There were 26 malignant and 15 benign tumours: 11 osteosarcomas , four Ewing’s sarcomas, four malignant fibrous histiocytomas (MFH) of pleomorphic type (undifferentiated high-grade pleomorphic sarcoma), two rhabdomyosarcomas, two low-grade myofibroblastic sarcomas,18et al.mRNA analyses were performed on all specimens according to the method described by Katou For immunohistochemistry, specimens were fixed in periodate–lysine 4% paraformaldehyde for 6–8 h at 4°C. After washing in phosphate-buffered saline, the fixed specimens were frozen. Primary antibodies were mouse monoclonal anti-FAP clone F19),,9 rat moThe US Food and Drug Administration-approved scoring system for HER2 protein was adopted for semiquantification of the localization patterns of FAP and DPP-IV because the immunoreactivity of both proteases along the cell membrane is similar to that of HER2.http://www.ncbi.nlm.nih.gov/) .Reverse transcriptase-polymerase chain reaction analyses identified the expression of mRNAs for both FAP and DPP-IV in all specimens examined . Direct To identify the cell types, immunohistochemistry was performed using pre-fixed frozen sections, not conventional paraffin-embedded sections, in order to attain more reliable sensitivity, and the results analysed using the semiquantification method . For malWe then analysed benign tumours to confirm the expression of FAP and DPP-IV. To analyse giant cell tumours, we subdivided the cellular components into giant cells, mononuclear round cells, and spindle-shaped fibroblastic cells. As summarized in The present study has clearly shown that fibroblast-like spindle tumour cells strongly express FAP and DPP-IV in low-grade myofibroblastic sarcoma, fibroblastic areas of osteosarcoma, MFH, non-ossifying fibroma, desmoid tumour, and chondroblastoma. The expression pattern is similar to that of activated fibroblasts and/or myofibroblasts in granulation tissue. In contrast, neither FAP nor DPP-IV is expressed in round or oval cells in Ewing’s sarcoma or rhabdomyosarcoma.Our results demonstrate the following two points. First, expression of FAP and DPP-IV is clearly independent of the malignant potential of the tumours. Second, it is rather related to the histogenesis of the tumour cells, i.e. positive tumour cells are related to activated fibroblasts and/or myofibroblasts. Myofibroblasts are activated fibroblasts that have some of the characteristics of smooth muscle cells, such as contractile properties.et al.et al.et al.We then considered the relationship to the malignant potential of the tumours, given that both FAP and DPP-IV are serine integral membrane proteinases. This suggests their involvement in cancer invasion and metastasis. However, as shown below, the situation is rather complicated. Cheng In addition to the above-mentioned expression in fibroblastic cells, DPP-IV is associated with another notable result of this study. Giant cells in giant cell tumours of bone were positive for DPP-IV. Giant cell tumours were formerly named osteoclastomas, and there has been debate concerning the relationship between giant cells in giant cell tumours and osteoclasts. Osteoclasts in normal bone tissue resemble macrophages in their histogenesis and function,In conclusion, both FAP and DPP-IV are expressed in tumour cells that are histogenetically related to activated fibroblasts and/or myofibroblasts, and their expression is not dependent on the malignancy of tumours. Our data suggest that FAP and DPP-IV can be used as markers of activated fibroblasts and/or myofibroblasts. DPP-IV may also be used as a marker of cells in the monocyte–macrophage lineage.


Racial and ethnic disparities in cervical cancer screening have been attributed to socioeconomic, insurance, and cultural differences. Our objective was to explore racial and ethnic differences in adherence to cervical cancer screening recommendations among female post-graduate physicians.We conducted a cross-sectional survey at one university hospital among a convenience sample of 204 female post-graduate physicians , examining adherence to United States Preventive Services Task Force cervical cancer screening recommendations, perception of adherence to recommendations, and barriers to obtaining care.Overall, 83% of women were adherent to screening recommendations and 84% accurately perceived adherence or non-adherence. Women who self-identified as Asian were significantly less adherent when compared with women who self-identified as white . Women who self-identified as East Indian were significantly less likely to accurately perceive adherence or non-adherence when compared to women who self-identified as white . Women who self-identified as Asian were significantly more likely to report any barrier to obtaining care when compared with women who self-identified as white  and there was a non-significant tendency toward women who self-identified as East Indian being more likely to report any barrier to obtaining care when compared with women who self-identified as white .Among a small group of insured, highly-educated physicians who have access to health care, we found racial and ethnic differences in adherence to cervical cancer screening recommendations, suggesting that culture may play a role in cervical cancer screening. Racial and ethnic disparities in cancer diagnosis and prognosis have been well documented. One explMany factors have been associated with decreased cervical cancer screening, including obesity,7 and olThe challenge of disentangling racial and ethnic differences in cervical cancer screening from differences related to socioeconomic status, education, insurance, or acculturation is immense. However, by studying cervical cancer screening among female post-graduate physicians, we had a unique opportunity to examine a group of insured, highly-educated women who have access to care and professional experience in cervical cancer and prevention. Physician preventive health care utilization has not been widely studied, but in instances when it has, some studies have found physicians to use preventive care services at high rates, -22 whileA cross-sectional survey was administered to female post-graduate physicians at Montefiore Medical Center, the University Hospital of the Albert Einstein College of Medicine, Bronx, NY, identified using a register of all trainees. All female post-graduate physicians were eligible for participation. Between May 2003 and January 2004, women were approached in-person by an investigator  to complete a self-administered, anonymous survey examining "health behaviors." Most housestaff were approached while alone in the hospital's medical library or cafeteria. After consenting to participate, housestaff were given an unmarked survey and an envelope that contained other completed surveys. The investigator stepped away for complete privacy and retrieved the envelope after the survey had been completed and placed with the others in the envelope, which required approximately five minutes. Completed surveys were not taken out of the envelope until the project's data collection phase was finished in order to further protect participants' privacy. Surveys were not distributed by electronic or inter-office mail because of response rate concerns. Institutional review board approval was obtained prior to the study.The survey assessed socio-demographic characteristics, including self-identified race/ethnicity, and had been piloted for readability but not validated. Multiple-choice questions were used to assess the following primary outcomes: 1) adherence to United States Preventive Services Task Force (USPSTF) recommendations for cervical cancer screening, 2) perception of personal adherence to recommendations, and 3) barriers to obtaining care among women who perceived themselves to be non-adherent to USPSTF recommendations. The following barrier responses were provided: no health insurance; no physician; not comfortable having Pap smear at own institution; no time; not sexually active; test inaccurate; and other (with space provided). Where appropriate, respondents could select more than one proposed response and additional space was provided for open-ended responses.The USPSTF gave cervical cancer screening an A recommendation, using indirect evidence to determine that screening women with a cervix within three years of sexual activity (or age 21) and at least every three years thereafter captures most of the benefit in terms of reducing morbidity and mortality. However,Data analysis was performed using SPSS version 11.5 . Associations between self-identified race/ethnicity and our primary outcomes were assessed using Pearson's Chi-Square and Fisher's Exact Test, depending upon the sample size for analysis, using self-identified white race as the reference group. Kappa was used to test for agreement beyond chance between adherence and perception of adherence to recommendations. All statistical tests were two-tailed.Among 393 female post-graduate physicians identified using a register of trainees, 206 were approached for participation. Response rate was > 99% (204 of 206); two declined participation, one citing urgent patient care responsibility and one citing no interest. Therefore, surveys were completed by 52% (204 of 393) of all female post-graduate physicians, representing a convenience sample.Mean age of respondents was 30 years; 53% self-identified themselves as white, 24% as Asian, 8% as African-American, 7% as East Indian, and 5% as Hispanic/Latina Table . Two eleOverall, 83% of women were adherent to USPSTF screening recommendations Table . Women wOverall, 84% of women accurately perceived personal adherence or non-adherence , which was reported by 50% of self-identified Asian women, 47% of self-identified East Indian women, and 30% of self-identified white women. Women who self-identified as Asian were more likely to report not feeling comfortable having a Pap smear at her workplace institution as a barrier to obtaining screening when compared with women who self-identified as white , while women who self-identified as East Indian were more likely to report not being sexually active as a barrier to obtaining screening when compared with women who self-identified as white . No women reported not having health insurance as a barrier to obtaining cervical cancer screening, nor did any women report cultural barriers to obtaining screening as an open-ended response .Among a small group of female post-graduate physicians at a single institution, racial and ethnic differences in current adherence to cervical cancer screening recommendations were found. Specifically, self-identified Asian women were less adherent to recommendations, while self-identified East Indian women less accurately perceived adherence or non-adherence to recommendations, erring in their perception of being non-adherent to recommendations when they were adherent. While barriers to obtaining screening were frequently reported, self-identified Asian women were more likely to report such barriers. Self-identified East Indian women also tended to be more likely to report such barriers, although this association was not statistically significant. Moreover, across all races and ethnicities, women reported barriers that are typical of physicians in post-graduate training: no time to schedule or keep an appointment.To our knowledge, this study is the first to examine cervical cancer screening among a racially and ethnically diverse group of post-graduate physicians, highly-educated women with professional experience in cervical cancer and prevention. Moreover, all housestaff during the time of the study were provided individual insurance policies at no cost and were able to access care either at the workplace (at on-site affiliated clinics) or outside of the workplace (at off-site affiliated or non-affiliated clinics). Because we found no differences in cervical cancer screening between self-identified white women and either African-American or Hispanic/Latina women, our study indirectly confirms prior research that demonstrated that adjusting for socioeconomic status and access to care explained observed differences in cervical cancer screening between white women and both African-American and Hispanic/Latina women.However, the observed difference in adherence to recommendations between self-identified Asian and white women, which has been shown in prior research,,16 suggeThere are several considerations in interpreting this study. First, the survey relied on open-ended responses for report of cultural barriers to care, such as orientation towards Eastern medicine or discomfort around sexuality. Perhaps physicians in post-graduate training were preoccupied by immediate barriers to obtaining care (lack of time) and did not consider reporting cultural barriers. Second, sub-samples of post-graduate female physicians by race/ethnicity were small, prohibiting conclusive comparisons, and only half of all post-graduate female physicians at this one large academic medical center were surveyed. Third, because we initiated the study in May 2003, our opportunity to survey the substantial number of post-graduate physicians in their final year of training was small . However, nearly all approached post-graduate physicians completed the survey, minimizing potential selection bias, and there was no difference in survey response by academic department. Fourth, our study is based on self-reported data. The tendency of respondents to over-report health promotion and disease-prevention activities is widely recognized. -29 HowevThis exploratory pilot study among insured, highly-educated physicians who have access to care found racial and ethnic differences in adherence to cervical cancer screening recommendations, suggesting that culture may play a role in cervical cancer screening. To address disparities in preventive service utilization, these results suggest that education, health literacy, insurance and access to care may be insufficient. Further work exploring cultural differences is needed.The authors declare that they have no competing interests.Drs. Ross and Nuñez-Smith were scholars in the Robert Wood Johnson Clinical Scholars Program at Yale University sponsored by the Robert Wood Johnson Foundation at times during their project involvement. Dr. Ross is currently supported by the Hartford Foundation and the Department of Veterans Affairs Health Services Research and Development Service project no. TRP-02-149. The views expressed in this article are those of the authors and do not necessarily represent the views of the Department of Veterans Affairs. Neither the Robert Wood Johnson nor Hartford Foundation had any role in the design or conduct of the study; collection, management, analysis or interpretation of the data; preparation, review or approval of the manuscript.JSR had full access to all the data in the study and takes responsibility for the integrity of the data and the accuracy of the analysis.Study concept and design: JSR, BAFAcquisition of data: JSR, BAFAnalysis and interpretation of data: JSR, MN-S, JRRDrafting of the manuscript: JSRCritical revision of the manuscript for important intellectual content: JSR, MN-S, BAF, JRRStatistical analysis: JSRAdministrative, technical, or material support: JSRStudy supervision: JSRThe pre-publication history for this paper can be accessed here:


Plasmodium berghei NK65, and the development of MA-ARDS was characterized by increased lung weight, edema, leukocyte infiltration and hemorrhages , characterized by pulmonary edema and hemorrhages. No efficient treatments are available for MA-ARDS and its pathogenesis remains poorly understood. To develop a new animal model for MA-ARDS, mice were infected with s Figure . The pulP. berghei ANKA, characterized by fulminant cerebral malaria. Hence, infection with P. berghei NK65 generates a broader time window to study the pathogenesis and to evaluate candidate treatments. The finding that high doses of dexamethasone cured MA-ARDS suggests that it might be more effective against MA-ARDS than it was in the clinical trials for cerebral malaria.We developed a novel model of MA-ARDS with many similarities to human MA-ARDS and without cerebral complications. This contrasts with the more classical model with


The adverse effects of tobacco abuse on cardiovascular outcomes are well-known. However, the impact of passive smoke exposure on angina status and therapeutic response is less well-established. We examined the impact of second-hand smoke (SHS) exposure on symptomatic improvement in patients with chronic ischemic coronary disease undergoing enhanced external counterpulsation (EECP).This observational study included 1,026 non-smokers (108 exposed and 918 not-exposed to SHS) from the Second International EECP Patient Registry. We also assessed angina response in 363 current smokers. Patient demographics, symptomatic improvement and quality of life assessment were determined by self-report prior and after EECP treatment.Non-smoking SHS subjects had a lower prevalence of prior revascularization (85% vs 90%), and had an increased prevalence of stroke (13% vs 7%) and prior smoking  compared to non-smokers without SHS exposure. Despite comparable degrees of coronary disease, baseline angina class, medical regimens and side effects during EECP, fewer SHS non-smokers completed a full 35-hour treatment course  compared to non-smokers without SHS. Compared to non-smokers without SHS, non-smoking SHS subjects had less angina relief after EECP , both higher than that achieved in current smokers (66%). By multivariable logistic regression, SHS exposure was an independent predictor of failure to symptomatic improvement after EECP among non-smokers .Non-smokers with SHS exposure had an attenuated improvement in anginal symptoms compared to those without SHS following EECP. Epidemiological evidence has unequivocally confirmed that active smoking is a risk factor for cardiovascular disease and the leading cause of preventable death ,2. The iEnhanced external counterpulsation (EECP) is a non-invasive treatment for chronic, refractory angina that involves the sequential inflation of three sets of lower-extremity cuffs during diastole, leading to increased venous return and cardiac output, systolic unloading, and augmentation of the coronary artery perfusion pressure. EECP has been shown to be effective in treating patients with chronic ischemic coronary artery disease using various measures, including improved functional class , reducedThere have been several emerging predictors of a lower likehood of benefit from EECP treatment including the presence of diabetes mellitus, prior surgical coronary revascularization and heart failure . MoreoveThe study group consisted of 1,026 non-smoking and 363 current smoking patients treated with EECP for refractory angina enrolled in the Second International EECP Patient Registry (IEPR-II). All patients signed informed written consent prior to entry into the Registry, and the Registry was approved by each center's institutional review board. The primary analyses were performed comparing the non-smokers without SHS exposure (n = 918) and non-smokers with SHS exposure (n = 108). The IEPR-II enrolled consecutive patients treated with EECP for chronic angina in 73 US centers between 2002 and 2004. Briefly, the IEPR-II collected patient demographics, medical history, and coronary artery disease status before and after initiation of EECP therapy. Data were collected prospectively on anginal status according to the Canadian Cardiovascular Society Classification (CCSC), anti-anginal medication use, quality of life and adverse clinical events. The present analysis includes patients who were enrolled from sites reporting ≥ 85% clinical follow-up. Since the IEPR aimed to collect data on as broad a range of patients as possible, the criteria for entry were only that the patient gave informed consent and had at least one hour of EECP treatment for chronic angina. The patients were interviewed by telephone 6 months after the last EECP treatment session, and yearly thereafter in order to record anginal status, quality of life, and adverse cardiac events . Non-smoData are presented as percentages for categorical variables or as mean values and standard deviations for continuous variables. Percentages and means are for patients with that data item recorded. Data were missing for fewer than 3% of each variable collected. Comparison of continuous variables between groups was analyzed by Wilcoxon rank tests and for categorical variables by chi-square, Fisher's exact test or Mantel-Henzel tests, as appropriate. Multivariable logistic regression analysis was used to determine independent predictors of failure to achieve reduction in angina. All baseline variables associated with the outcome with a p-value of < 0.2 were entered into the preliminary model. A backward selection procedure was used to determine independent, statistically significant predictors. Two-tailed p-values < 0.05 were defined as significant.Of the 1,026 non-smoking patients who underwent EECP treatment for angina, 108 (10.5%) reported current SHS exposure. Demographic and clinical characteristics, such as the prevalence of hypertension, diabetes, prior myocardial infarction, heart failure, and anginal class, were similar among the two groups of non-smokers . The rate of major adverse cardiovascular events  during the course of EECP therapy was 5.6% for the SHS group, compared to 2.8% for the non-SHS group (p = 0.12). However, there was a higher incidence of unstable angina during EECP in the SHS group (8.3%) compared to the non-SHS non-smoker group . Other adverse events occurring during the EECP treatment period occurred at a similar rate  both higher than the 66% angina reduction achieved by the current smoking group , age ≤ 55 years , and mild angina (CCSC I/II) pre-EECP .In this large cohort of patients with refractory angina undergoing EECP, SHS exposure was an independent predictor of failure to achieve a reduction in angina class after treatment among non-smokers. Only 68% of those non-smoking subjects with SHS had a reduction in angina class, compared to 79% of the non-smokers without SHS exposure. This low level of angina reduction is comparable to that achieved among current cigarette smokers (66%). Moreover, the SHS group had a higher rate of adverse unstable angina cardiac events during the course of EECP compared to non-smokers without SHS exposure.Prior clinical studies observed impaired endothelium-dependent vasorelaxation, the earliest sign of endothelial dysfunction, in smokers -27. PassThe agents and pathways responsible for these functional changes of endothelial function have not yet been completely elucidated, but they may be related, at least in part, to the inactivation of nitric oxide (NO) . Animal EECP has been shown to increase c-GMP production, which promotes vascular smooth muscle tone and improves arterial function . EECP alA limitation of this analysis was the assessment of SHS exposure. By defining SHS exposure as only house-hold exposure, we may have underestimated the proportion of subjects with occupational SHS. However, the vast majority of patients undergoing EECP have severe angina, and are not employed outside the home. Self-reporting of the SHS status may be a limitation. There may be other unmeasured confounding variables, such as obesity, poor diet, and physical inactivity, which may be associated with SHS. Our database was not able to capture these potential confounding variables. The IEPR-2 Registry was designed as a cohort study of patients undergoing EECP and provides a rich data set for exploration. As with many analyses of the Registry data, the SHS hypothesis was not pre-specified; this should be kept in mind in the interpretation of the findings.In conclusion, SHS may reduce the anti-anginal effects of EECP among non-smokers. This reduced success rate of EECP in SHS non-smokers is comparable to that seen in current cigarette smokers. These findings are consistent with studies showing that SHS exposure reduces nitric oxide production, which is an important mechanism for the clinical benefit seen with EECP.ARIC: Atherosclerosis Risk in Communities; CABG: Coronary artery bypass graft surgery; CCSC: Canadian Cardiovascular Society Classification; CO: Carbon Monoxide; CVD: Cardiovascular disease; EECP: Enhanced external counterpulsation; IEPR-II: Second International EECP Patient Registry; MACE: Major adverse cardiovascular events; NO: Nitric oxide; SHS: second-hand smokeThe IEPR-2 is sponsored by Vasomedical, Inc., Westbury, NY. Drs. Michaels, Kennard, and Kelsey have performed consulting for Vasomedical Inc., the manufacturer of EECP. Dr. Michaels has also participated on the speaker's bureau for Vasomedical Inc.SE drafted the manuscript. EK, SF, and AM conceived of the study, and participated in its design and coordination and helped to draft the manuscript. All authors read and approved the final manuscript.The pre-publication history for this paper can be accessed here:


Cowpox virus (CPXV) from the ulcerative eyelid lesions of a 14-year-old girl, who had cared for a clinically ill wild rat that later died. CPXV isolated from the rat (Rattus norvegicus) showed complete homology with the girl’s virus. Our case is the first proven rat-to-human transmission of cowpox.We isolated  A 14-year-old girl was admitted to the hospital with ulcerated nodules on her upper lip and her left lower eyelid and several molluscum-like-lesions on her right eyelids . She wasOrthopoxvirus. Serum immunoglobulin (Ig) M antibodies to the viral isolate and to Vaccinia virus were detected upon admission. The virus was identified as Cowpoxvirus (CPXV) by polymerase chain reaction (PCR) and sequence analysis   . The ratHuman CPXV is a rare zoonosis, which is transmitted as a result of contact with animals.Parapoxvirus). CPXV is almost always a self-limiting disease in immunocompetent hosts.Usually localized at the site of inoculation, CPXV lesions progress from a papule through vesiculation and pustulation into an ulcerative nodule, which is covered with an elevated border with a black eschar. Ulcera heal with scar formation. Differential clinical diagnosis of CPXV includes herpes virus infection, anthrax, and orf , wood mice (Apodemus sylvaticus) and field voles (Microtus agretis)  . Seroprefections ,7. Physi


This paper describes a GC/MS system capable of performing Volatile Organic Analysis on liquids, solids, and air. When combined with a syringe auto-injector, the system is completely automated for both volatile and semi-volatile analyses. An OI Analytical Model 4551 Vial Multisampler and an OI Analytical DPM-16 Multisampler are interfaced and then connected to an OI Analytical Model 4560 Sample Concentrator, an HP Model 5971 MSD, an HP Model 7673 Auto-Injector, and an HP Model 5890 Series II GC to form a multi-tasking GC/MS system. This system is shown to allow greater versatility in the laboratory.