Rapid proteome analysis of bronchoalveolar lavage samples of lifelong smokers and never-smokers by micro-scale liquid chromatography and mass spectrometry

BACKGROUND: The aim of this study was to determine whether relative qualitative and quantitative differences in protein expression could be related to smoke exposure or smoke-induced airway inflammation. We therefore explored and characterized the protein components found in bronchoalveolar lavage (BAL) fluid sampled from either lifelong smokers or never-smokers. METHODS: BAL fluid samples obtained by bronchoscopy from 60-year-old healthy never-smokers (n = 18) and asymptomatic smokers (n = 30) were analyzed in either pooled or individual form. Initial global proteomic analysis used shotgun digestion approaches on unfractionated BAL fluid samples (after minimal sample preparation) and separation of peptides by gradient (90-min) liquid chromatography (LC) coupled with on-line linear ion trap quadropole mass spectrometry (LTQ MS) for identification and analysis. RESULTS: LTQ MS identified 481 high- to low-abundance proteins. Relative differences in patterns of BAL fluid proteins in smokers compared with never-smokers were observed in pooled and individual samples as well as by 2-dimensional gel analysis. Gene ontology categorization of all annotated proteins showed a wide spectrum of molecular functions and biological processes. CONCLUSIONS: The described method provides comprehensive qualitative proteomic analysis of BAL fluid protein expression from never-smokers and from smokers at risk of developing chronic obstructive pulmonary disease. Many of the proteins identified had not been detected in previous studies of BAL fluid; thus, the use of LC-tandem MS with LTQ may provide new information regarding potentially important patterns of protein expression associated with lifelong smoking.     

Adherence to a daily flossing regimen in university students: effects of planning when, where, how and what to do in the face of barriers

OBJECTIVES: Regular (daily) dental flossing is recommended for preventing oral diseases, but adherence is unsatisfactory. Social cognitive theory (SCT) specifies determinants of dental flossing: cognitions about risk, positive and negative outcome expectations and the perceived ability to perform behaviour predict motivation, which in turn predicts behaviour. Recent research suggests that motivation alone may not suffice to predict behaviour, and proposes if-then-planning. This study aims to predict flossing adherence from social cognitive variables and planning. MATERIAL AND METHODS: Questionnaire data from 157 non-dental university students on flossing, SCT variables and planning were gathered at three measurement points over 6 weeks. Residual floss was used to validate behaviour self-reports. RESULTS: Social cognitive variables and planning correlated significantly with flossing at all times. Discriminant function analysis suggests that after controlling for Time 1 flossing, planning Time 2 (Wilk's lambda=0.77; p<0.01) is more important in discriminating between adherent and non-adherent participants at Time 3 than Time 1 social cognitive measures. Regression analyses confirmed this result with planning as only predictor of flossing change (p<0.05). CONCLUSIONS: These results suggest targeting planning in interventions to increase compliance with flossing recommendations. Implications for such interventions are discussed.     

Mutations in the calcium-related gene IL1RAPL1 are associated with autism

In a systematic sequencing screen of synaptic genes on the X chromosome, we have identified an autistic female without mental retardation (MR) who carries a de novo frameshift Ile367SerfsX6 mutation in Interleukin-1 Receptor Accessory Protein-Like 1 (IL1RAPL1), a gene implicated in calcium-regulated vesicle release and dendrite differentiation. We showed that the function of the resulting truncated IL1RAPL1 protein is severely altered in hippocampal neurons, by measuring its effect on neurite outgrowth activity. We also sequenced the coding region of the close related member IL1RAPL2 and of NCS-1/FREQ, which physically interacts with IL1RAPL1, in a cohort of subjects with autism. The screening failed to identify non-synonymous variant in IL1RAPL2, whereas a rare missense (R102Q) in NCS-1/FREQ was identified in one autistic patient. Furthermore, we identified by comparative genomic hybridization a large intragenic deletion of exons 3-7 of IL1RAPL1 in three brothers with autism and/or MR. This deletion causes a frameshift and the introduction of a premature stop codon, Ala28GlufsX15, at the very beginning of the protein. All together, our results indicate that mutations in IL1RAPL1 cause a spectrum of neurological impairments ranging from MR to high functioning autism.     

Conceptions of acculturation: a review and statement of critical issues

This article reviews evidence for re-conceptualizing acculturation status and acculturation process in health care research with United States (U.S.) Latino populations. Prior literature on acculturation has focused on: (a) acculturative change towards the dominant culture, (b) acculturation as it occurs with Mexican Americans, and (c) language as the principal component of acculturation. We review empirically based health research on acculturation and propose an ecodevelopmental framework for better understanding the process of acculturation. We then offer recommendations that may assist public health researchers, policymakers and program developers in better understanding "real world" acculturation. This includes understanding acculturation within this ecodevelopmental context for a more complete understanding of the acculturation process and its influences on health-related behaviors, with aims of reducing or eliminating health disparities in Latino populations.     

Influence of pyridoxal phosphate in measuring aminotransferases activities in patients with viral hepatitis

Effect of a pyridoxal phosphate (PP) supplementation of reagents used for ALT and AST measurement was studied in serum of 20 patients suffering from viral hepatitis. Measurements of enzyme activities were carried out at 37 degrees C, using an automate (AU 600, Olympus). Significant differences (p < 0.0001) were observed both for ALT and AST, meanwhile they were more marked for ALT than for AST. This difference was associated with a strong interindividual variability regarding PP activation effect on ALT. In conclusion, aminotransferase measurements should be carried out with a reagent supplemented with PP, when the enzyme activity is used to evaluate a cytolysis. The same recommendation applies when ALT results are integrated into various combinations developed for the evaluation of liver status.     

Mitogenic responses of canine peripheral blood lymphocytes to staphylococcal protein A

In order to produce long term lymphoid cell cultures from canine lymphocytes of known histocompatibility antigen specificities, mitogenic responses to staphylococcal protein A (SpA) were examined and compared with those of phytohaemagglutinin (PHA) and concanavalin A (Con A). SpA was found to be the strongest mitogen tested with significant responses to concentrations as low as 31 ng/ml. There was a decrease in responsiveness above optimal mitogen concentrations with SpA and PHA. Peak responses were observed at lower concentrations for longer incubation times. PHA showed a rapid fall off in thymidine uptake below optimal concentrations whereas the SpA dose-response curve was less steep and a shoulder or secondary peak of activity was observed at low SpA concentrations in some cases.Continuous SpA stimulation of lymphocyte cultures resulted in an initial period of cell proliferation followed usually by a second period of cell proliferation around week 7 of culture. To date, viable cell cultures have been maintained for up to 12 weeks in vitro.SpA lymphoblast cultures behave normally in microcytotoxicity tests for serologically defined DLA histocompatibility antigens and remain functional in natural killer (NK) and PHA induced cell mediated cytotoxic reactions against ⁵¹Cr-labelled tumour target cells but were not themselves susceptible as target cells for NK activity.     

Enzyme calibrators: principle and practical use

Results of catalytic activities of enzymes are highly dependent on the measurement procedures and on local conditions. Thus, only poorly marked improvement of interlaboratory comparability of results have been observed in clinical enzymology. To solve this problem, SFBC and IFCC have proposed to use "validated enzyme calibrators". Standardised operating procedures adapted to 37 C have been developed by IFCC for the most commonly used enzymes in clinical chemistry, and will be soon published. Reference materials which have been certified with these SOPs can be used as calibrators for a set of measurement methods which exhibit the same analytical specificity. Calibrators must be commutable, a property that must be checked experimentally. It is possible to produce stable and commutable materials for the calibration of a set of methods. Interest of this approach has been demonstrated for several enzymes. Results of two studies presented here show that the comparison of results to the upper limit of reference ranges does not improve the interlaboratory comparability of results in contrast to the calibration of different methods by a common calibrator which allowed to reach an interlaboratory CV close to 4% for ALT and gammaGT.