Project description:Atmospheric particulate matter (PM) is a recognized risk factor for the global burden of disease in human populations. We are presenting here the application of toxicogenomics in the evaluation of the toxic effects of organic content of atmospheric particle matter (PM), from urban and rural environments (city of Barcelona and village of La Pobla, NE Spain), using the developing zebrafish embryo. The main goal is to identify the metabolic pathways involved in the adverse effects observed in zebrafish embryos exposed to PM organic content from urban and rural environments, also allowing the selection of genes of interest that are differentially expressed. The relevance of particle size to the PM toxicity is also addressed. Indeed, the zebrafish embryos were exposed to PM of aerodynamic diameter larger than 7.2 μm and smaller than 0.5 μm (PM10 and PM0.5, respectively). PM0.5 concentrated biological and toxic activities linked to organic substances. Transcriptomic analyses showed strong induction of the AhR signalling pathway (a.k.a. dioxin-like activity) for embryos exposed to both rural and urban extracts, correlating with the concentrations of PAHs. Urban extracts, with strong contribution of traffic emissions, specifically de-regulated oxidative stress-related genes, as well as pancreatic and eye-lens specific genes, two organs known to be affected by air pollution in humans. Exposure to rural extracts, with high contribution of wood burning emissions, affected genes implicated in basic cellular functions, in agreement with their strong embryotoxicity. Extracts from rural and urban samples elicited both common and specific transcriptome responses, suggesting different potentially adverse outcomes depending on PM source and composition. The authors thank the financial support of the Spanish Ministry (project TEA-PARTICLE, grant number CGL2011-29621) and the Portuguese Foundation for Science and Technology for the doctoral grant of Sofia R. Mesquita (SFRH/BD/80710/2011) funded by the Program POPH-QREN through the Portuguese Ministry of Education and Science and the European Social Fund, and support through project PEst-C/MAR/LA0015/2013.

Project description:Atmospheric particulate matter (PM) is a recognized risk factor for the global burden of disease in human populations. We are presenting here the application of toxicogenomics in the evaluation of the toxic effects of organic content of atmospheric particle matter (PM), from urban and rural environments (city of Barcelona and village of La Pobla, NE Spain), using the developing zebrafish embryo. The main goal is to identify the metabolic pathways involved in the adverse effects observed in zebrafish embryos exposed to PM organic content from urban and rural environments, also allowing the selection of genes of interest that are differentially expressed. The relevance of particle size to the PM toxicity is also addressed. Indeed, the zebrafish embryos were exposed to PM of aerodynamic diameter larger than 7.2 Î¼m and smaller than 0.5 Î¼m (PM10 and PM0.5, respectively). PM0.5 concentrated biological and toxic activities linked to organic substances. Transcriptomic analyses showed strong induction of the AhR signalling pathway (a.k.a. dioxin-like activity) for embryos exposed to both rural and urban extracts, correlating with the concentrations of PAHs. Urban extracts, with strong contribution of traffic emissions, specifically de-regulated oxidative stress-related genes, as well as pancreatic and eye-lens specific genes, two organs known to be affected by air pollution in humans. Exposure to rural extracts, with high contribution of wood burning emissions, affected genes implicated in basic cellular functions, in agreement with their strong embryotoxicity. Extracts from rural and urban samples elicited both common and specific transcriptome responses, suggesting different potentially adverse outcomes depending on PM source and composition. The authors thank the financial support of the Spanish Ministry (project TEA-PARTICLE, grant number CGL2011-29621) and the Portuguese Foundation for Science and Technology for the doctoral grant of Sofia R. Mesquita (SFRH/BD/80710/2011) funded by the Program POPH-QREN through the Portuguese Ministry of Education and Science and the European Social Fund, and support through project PEst-C/MAR/LA0015/2013. The PM filter samples used in the present study were PM10 and PM0.5 from the urban site - total of 4 samples from 2 sampling months; and PM0.5 from the rural site - total of 2 samples from 2 sampling months (PM10 samples from the rural site were not tested due to their very low concentration in organic compounds, and insignificant biological activity, previously measured). Sampling occurred during the cold periods of the year 2012/2013. The organic fraction of PM samples was extracted by sonication using a mixture of dichloromethane:methanol. Then, the extracts were filtered, evaporated and re-dissolved in Dimethyl sulfoxide (DMSO). Zebrafish fertilized eggs were exposed to PM organic extracts (0.2% DMSO) from the urban and rural sites from 24 to 120h post-fertilization. Total RNA was isolated from whole embryos (pools of 20 individuals), using Trizol reagent protocol (Invitrogen Life Technologies, Carlsberg, CA). The microarray study was performed using the Agilent Two-colour D. rerio Oligo Microarray v3 platform, following the Agilent Microarray â?? Based Gene Expression Analysis protocol. Three biological replicates were performed for each PM sample and controls. Biological replicates, a technical replicate and a self-to-self, were simultaneously amplified and labelled using Cyanine 3 (Cy3) and Cyanine 5 (Cy5) dye. After cRNA purification (RNeasy Kit, Quiagen GmbH, Hilden, Germany), the cRNA concentration and labelling were quantified in the NanoDrop spectrophotometer, obtaining incorporation rates in the range of 15-20pmol of cyanide dye/Âµl cRNA and yield values > 0.825ug, as recommended. cRNA was fragmented and hybridized in 4x44K arrays, for 17h at 65oC. After the hybridization the array slides were washed, and immediately scanned using Microarray Scanner Agilent G2505C system. Data was extracted using Agilent Feature Extraction Software v10.5.1.1, and the quality of microarray data was evaluated using the Quality Control report provided by Agilent Software. Based on the microarray data analysis, 22 primers were selected and designed using Primer Express 2.0 software (Applied Biosystems, Foster City, CA). Genes were quantified by real-time PCR to validate the microarray data, and a subset of 13 genes were further selected by their robust behaviour in the validation assays, allowing to differentiate the toxic potential of PM from urban and rural sources.

Project description:IntroductionLow back pain (LBP) is highly prevalent in Nigeria and is more devastating in rural Nigeria due to adverse living and working conditions, reinforced by maladaptive illness beliefs. There is a need to develop measures for assessing such beliefs in this population. This study aimed to cross-culturally adapt the Fear Avoidance Beliefs Questionnaire (FABQ) and test its psychometric properties in mixed rural and urban Nigerian populations with chronic LBP.MethodsTranslation, cultural adaptation, test-retest, and cross-sectional psychometric testing. FABQ was forward and back translated by clinical/non-clinical translators. A review committee evaluated the translations. Twelve people with chronic LBP in a rural Nigerian community pre-tested the questionnaire. Cronbach's alpha assessing internal consistency; intra-class correlation coefficient and Bland-Altman plots assessing test-retest reliability; and minimal detectable change were investigated in a convenient sample of 50 chronic low back pain sufferers in rural and urban Nigeria. Construct validity was examined using Pearson's correlation analyses with the eleven-point box scale and Igbo Roland Morris Disability Questionnaire (Igbo-RMDQ), and exploratory factor analysis in a random sample of 200 adults with chronic low back pain in rural Nigeria. Ceiling and floor effects were investigated in all samples.ResultsAmendments allowed interviewer-administration. Item 8 was modified to 'I have a compensation or gains I get from having my pain' as there is no benefit system in Nigeria. Igbo phrase for 'physical activity' could also mean 'being active', 'moving the body' or 'moving about' and was used in the items with 'physical activity'. The Igbo-FABQ had good internal consistency (α = 0.80-0.86); intra class correlation coefficients (ICC = 0.71-0.72); standard error of measurements (3.21-7.40) and minimal detectable change (8.90-20.51). It correlated moderately with pain intensity and disability, with a two-factor structure and no floor and ceiling effects.ConclusionsIgbo-FABQ is valid, reliable, and can be used clinically and for research.

Project description:IntroductionNegative attitudes and beliefs about low back pain (LBP) can lead to reduced function and activity and consequently disability. One self-report measure that can be used to assess these negative attitudes and beliefs and to determine their predictive nature is the Back Beliefs Questionnaire (BBQ). This study aimed to translate and cross-culturally adapt the BBQ into Hausa and assess its psychometric properties in mixed urban and rural Nigerian populations with chronic LBP.MethodsThe BBQ was translated and cross-culturally adapted into Hausa (Hausa-BBQ) according to established guidelines. To assess psychometric properties, a consecutive sample of 200 patients with chronic LBP recruited from urban and rural clinics of Nigeria completed the questionnaire along with measures of fear-avoidance beliefs, pain catastrophizing, functional disability, physical and mental health, and pain. One hundred of the 200 patients completed the questionnaire twice at an interval of 7-14 days to assess test-retest reliability. Internal construct validity was assessed using exploratory factor analysis, and external construct validity was assessed by examining convergent, divergent, and known-groups validity. Reliability was assessed by calculating internal consistency (Cronbach's α), intraclass correlation coefficients (ICC), standard error of measurement (SEM), minimal detectable change at 95% confidence interval (MDC95), and limits of agreement using Bland-Altman plots. Reliability (ICC, SEM and MDC95) was also assessed separately for rural and urban subgroups.ResultsThe factor analysis revealed a four-factor solution explaining 58.9% of the total variance with the first factor explaining 27.1%. The nine scoring items loaded on the first factor hence supporting a unidimensional scale. The convergent and divergent validity were supported as 85% (6:7) of the predefined hypotheses were confirmed. Known-groups comparison showed that the questionnaire discriminated well for those who differed in education (p < 0.05), but not in age (p > 0.05). The internal consistency and ICC (α = 0.79; ICC = 0.91) were adequate, with minimal SEM and MDC95 (1.9 and 5.2, respectively). The limits of agreements were -5.11 to 5.71. The ICC, SEM and MDC95 for the urban and rural subgroups were comparable to those obtained for the overall population.ConclusionsThe Hausa-BBQ was successfully adapted and psychometrically sound in terms of internal and external construct validity, internal consistency, and test-retest reliability in mixed urban and rural Hausa-speaking populations with chronic LBP. The questionnaire can be used to detect and categorize specific attitudes and beliefs about back pain in Hausa culture to prevent or reduce potential disability due to LBP.

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Public beliefs on antibiotics and symptoms of respiratory tract infections among rural and urban population in Poland: a questionnaire study.

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