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Phytogenic compounds may influence salivation or salivary properties. However, their effects on the bovine salivary proteome have not been evaluated. We investigated changes in the bovine salivary proteome due to transition from forage to high-concentrate diet, with and without supplementation with a phytogenic feed additive. Eight non-lactating cows were fed forage, then transitioned to a 65% concentrate diet (DM basis) over a week. Cows were control (n = 4, CON) or supplemented with a phytogenic feed additive (n = 4, PHY). Proteomic analysis was conducted using liquid chromatography coupled with mass spectrometry. We identified 1233 proteins; 878 were bovine proteins, 189 corresponded to bacteria, and 166 were plant proteins. Between forage and high-concentrate, 139 proteins were differentially abundant (P 0.05), with 48 proteins having a log2FC difference |2|. The salivary proteome reflected shifts in processes involving nutrient utilization, body tissue accretion, and immune response. Between PHY and CON, 195 proteins were differently abundant (P 0.05), with 37 having a log2FC difference |2|; 86 proteins were increased by PHY, including proteins involved in smell recognition. Many differentially abundant proteins correlated (r |0.70|) with salivary bicarbonate, total mucins or pH. Results provide novel insights into the bovine salivary proteome using a non-invasive approach, and the association of specific proteins with major salivary properties influencing rumen homeostasis. SIGNIFICANCE: Phytogenic compounds may stimulate salivation due to their olfactory properties, but their effects on the salivary proteome have not been investigated. We investigated the effect of high-concentrate diets and supplementation with a phytogenic additive on the salivary proteome of cows. We show that analysis of cows' saliva can be a non-invasive approach to detect effects occurring not only in the gut, but also systemically including indications for gut health and immune response. Thus, results provide unique insights into the bovine salivary proteome, and will have a crucial contribution to further understand animal response in terms of nutrient utilization and immune activity due to the change from forage to a high-energy diet. Additionally, our findings reveal changes due to supplementation with a phytogenic feed additive with regard to health and olfactory stimulation. Furthermore, findings suggest an association between salivary proteins and other components like bicarbonate content.

BackgroundWe examined quit attempts, use of cessation assistance, quitting beliefs and intentions among smokers who participated in the 2018 International Tobacco Control (ITC) Europe Surveys in eight European Union Member States (England, Germany, Greece, Hungary, the Netherlands, Poland, Romania and Spain).MethodsCross-sectional data from 11 543 smokers were collected from Wave 2 of the ITC Six European Country (6E) Survey (Germany, Greece, Hungary, Poland, Romania and Spain—2018), the ITC Netherlands Survey (the Netherlands—late 2017) and the Four Countries Smoking and Vaping (4CV1) Survey (England—2018). Logistic regression was used to examine associations between smokers’ characteristics and recent quit attempts.ResultsQuit attempts in the past 12 months were more frequently reported by respondents in the Netherlands (33.0%) and England (29.3%) and least frequently in Hungary (11.5%), Greece (14.7%), Poland (16.7%) and Germany (16.7%). With the exception of England (35.9%), the majority (56–84%) of recent quit attempts was unaided. Making a quit attempt was associated with younger age, higher education and income, having a smoking-related illness and living in England. In all countries, the majority of continuing smokers did not intend to quit in the next 6 months, had moderate to high levels of nicotine dependence and perceived quitting to be difficult.ConclusionsApart from England and the Netherlands, smokers made few quit attempts in the past year and had low intentions to quit in the near future. The use of cessation assistance was sub-optimal. There is a need to examine approaches to supporting quitting among the significant proportion of tobacco users in Europe and increase the use of cessation support as part of quit attempts

Background The EUREST-PLUS ITC Europe surveys aim to evaluate the impact of the European Union’s Tobacco Products Directive (EU TPD) implementation within the context of the WHO FCTC. This article describes the methodology of the 2016 (Wave 1) and 2018 (Wave 2) International Tobacco Control 6 European (6E) Country Survey in Germany, Greece, Hungary, Poland, Romania and Spain; the England arm of the 2016 (Wave 1) and 2018 (Wave 2) ITC 4 Country Smoking and Vaping (4CV) Survey; and the 2016 (Wave 10) and 2017 (Wave 11) ITC Netherlands (NL) Survey. All three ITC surveys covering a total of eight countries are prospective cohort studies with nationally representative samples of smokers. Methods In the three surveys across the eight countries, the recruited respondents were cigarette smokers who smoked at least monthly, and were aged 18 and older. At each survey wave, eligible cohort members from the previous waves were retained, regardless of smoking status, and dropouts were replaced by a replenishment sample. Results Retention rates between the two waves of the ITC 6E Survey by country were 70.5% for Germany, 41.3% for Greece, 35.7% for Hungary, 45.6% for Poland, 54.4% for Romania and 71.3% for Spain. The retention rate for England between ITC 4CV1 and ITC 4CV2 was 39.1%; the retention rates for the ITC Netherlands Survey were 76.6% at Wave 10 (2016) and 80.9% at Wave 11 (2017). Conclusion The ITC sampling design and data collection methods in these three ITC surveys allow analyses to examine prospectively the impact of policy environment changes on the use of cigarettes and other tobacco products in each country, to make comparisons across the eight countries.

The glomerular basement membrane (GBM) and extra-cellular matrix (ECM) are essential to maintain a functional interaction between the glomerular podocytes and the fenestrated endothelial cells in the formation of the slit diaphragm for the filtration of blood. Dysregulation of ECM homeostasis can cause Focal segmental glomerulosclerosis (FSGS). Despite this central role, alterations in ECM composition during FSGS have not been analyzed in detail yet. Here, we characterized the ECM proteome changes in miR-193a-overexpressing mice, which suffer from FSGS due to suppression of Wilms’ tumor 1 (WT1). By mass spectrometry we identified a massive activation of the acute phase response, especially the complement and fibrinogen pathways. Several protease inhibitors (ITIH1, SERPINA1, SERPINA3) were also strongly increased. Complementary analysis of RNA expression data from both miR-193a mice and human FSGS patients identified additional candidate genes also mainly involved in the acute phase response. In total, we identified more than 60 dysregulated, ECM-associated genes with potential relevance for FSGS progression. Our comprehensive analysis of a murine FSGS model and translational comparison with human data offers novel targets for FSGS therapy.

Macadamia nut is an increasingly popular food item of a healthy diet. However, macadamia nut is also a potent allergenic food. To date, there is little information about the allergenic proteins involved. In this study, using sera from macadamia nut allergic individuals, four IgE-binding proteins were detected. Their identities were determined by tandem mass spectrometry with de novo sequencing. Three IgE-reactive proteins, the vicilin Mac i 1, the legumin Mac i 2 and the antimicrobial peptide 2a/Mac i 1 (28-76) were purified from the nut while the non-specific lipid transfer protein was produced as a recombinant in Pichia pastoris. IgE-binding assays using sera from well-characterized groups of tree nut and/or peanut allergic patients revealed that the allergens were mainly recognized by sera from macadamia nut allergic individuals. Hence, these newly discovered allergens will enable molecular diagnostics to identify patients at high risk of macadamia nut allergy.

Devil's claw is used for the treatment of inflammatory symptoms and degenerative disorders in horses since many years, but without the substantive pharmacokinetic data. The pharmacokinetic parameters of harpagoside, the main active constituent of Harpagophytum procumbens DC ex Meisn., were evaluated in equine plasma after administration of Harpagophytum extract FB 8858 in an open, single‐dose, two‐treatment, two‐period, randomized cross‐over design. Six horses received a single dose of Harpagophytum extract, corresponding to 5 mg/kg BM harpagoside, and after 7 days washout period, 10 mg/kg BM harpagoside via nasogastric tube. Plasma samples at certain time points (before and 0–24 hr after administration) were collected, cleaned up by solid‐phase extraction, and harpagoside concentrations were determined by LC‐MS/MS using apigenin‐7‐glucoside as internal standard. Plasma concentration‐time data and relevant parameters were described by noncompartmental model through PKSolver software. Harpagoside could be detected up to 9 hr after administration. C max was found at 25.59 and 55.46 ng/ml, t 1/2 at 2.53 and 2.32 hr, respectively, and t max at 1 hr in both trials. AUC 0–inf was 70.46 and 117.85 ng hr ml−1, respectively. A proportional relationship between dose, C max and AUC was observed. Distribution (V z/F) was 259.04 and 283.83 L/kg and clearance (CL/F) 70.96 and 84.86 L hr−1 kg−1, respectively. Treatment of horses with Harpagophytum extract did not cause any clinically detectable side effects.

Dichlorodiphenyltrichloroethane (DDT), a persistent organochlorine pesticide, has been linked to adverse biological effects in organisms. However, there is limited knowledge about its toxic effects on marine organisms and the underlying molecular mechanisms. This study investigated the toxic effects of DDT in the hooded oyster Saccostrea cucullata. The oysters were exposed to DDT at concentrations of 0, 10, 50, 100, 500, 1000 and 2000 µg/L for 96 h and the LC50 (96 h) was 891.25 µg/L. Two sublethal concentrations (10 and 100 µg/L) were used to investigate the histopathological effects and the proteome response. Histopathological results showed that DDT caused the alteration of mantle tissue. This included the induction of mucocytes in the epithelium and the inflammatory effect in the connective tissue indicated by the enlargement of blood sinus and hemocyte aggregation within the sinus. Proteomic results showed that, amongst approximately 500 protein spots that were detected across 2DE gels, 51 protein spots were differentially expressed (P 1.2). Of these, 29 protein spots were identified by LC-MS/MS. These included 23 up-regulated, 5 down-regulated and 1 fluctuating spots. Thus, we observed that stress response and cytoskeletal proteins are the central targets of DDT action. Furthermore, DDT alters the expression of proteins involved in energy metabolism, calcium homeostasis and other proteins of unknown function. Additionally, proteomic results clearly elucidated the molecular response of the histopathological changes which were driven by the alteration of cytoskeletal proteins. Our results improve the current knowledge of toxicity of the DDT to histology and molecular response of oyster proteome to DDT exposure. In addition, histopathological changes will be beneficial for the development of an appropriate guideline for health assessment of this species in ecotoxicological context.

Background: Economic recessions may influence illegal drug use via different mechanisms, for example increased use due to more psychological distress or decreased use due to lower incomes and purchasing power. This paper reviews the literature on how economic recessions and unemployment affect the use of illegal drugs among adults. Methods: We conducted a systematic realist literature review, which is an explanatory method that aims to understand underlying mechanisms that connect an event to an outcome in a specific context. A systematic search was performed in EconLit, Embase, Medline, PsycINFO, SocIndex, and Web of Science for studies examining mechanisms explaining how recessions or unemployment affect illegal drug use. Results: We synthesized 28 studies published between 1990 and 2015. Most evidence (17 studies) was found for the counter-cyclical mechanism that recessions and unemployment increase psychological distress, which increases drug use. Mainly supportive evidence for this mechanism was found in several high quality studies, in different contexts, and in a diverse number of countries and samples. In contrast, decreased income did not seem to decrease drug use (10 studies). Little evidence was available on the non-working time mechanism (4 studies) and the social exclusion mechanism (5 studies). Most of the studies that did examine these latter mechanisms confirmed the hypothesized counter-cyclical associations. Conclusion: The current evidence is in line with the hypothesis that drug use increases in times of recession because unemployment increases psychological distress which increases drug use. During times of recession, psychological support for those who lost their job and are vulnerable to drug use (relapse) is likely to be important. (C) 2017 The Author(s). Published by Elsevier B.V.

Introduction:The relationship between work loss and smoking has not been studied extensively, and underlying factors are often not examined. The aim of this study was to test two hypotheses. First, work loss is associated with greater intention to quit and more likelihood of smoking cessation, and this relationship is moderated by a decrease in income. Second, work loss is associated with lower quit intention and lower rates of smoking cessation, and this relationship is moderated by an increase in psychological distress.Methods:We used pooled data from three countries participating in the ITC Project: France, Germany and the Netherlands (n=2712). We measured unemployment, income and psychological distress at two consecutive survey waves, and calculated changes between survey waves. We first conducted multiple logistic regression analyses to examine the association between work loss and smoking cessation behavior. Next, we added income decrease and psychological distress increase to the models. Finally, we added interaction terms of work loss by income decrease and work loss by distress increase to the model.Results:Work loss was not associated with quit intention, quit attempts, and quit success. When income decrease and psychological distress increase were added to the model, we found a positive association between distress increase and quit attempts. The interactions, however, were not statistically significant.Conclusions:Our results indicate that smokers who become unemployed and face a decrease in income are not less likely to quit smoking than smokers who are employed.

This study examined to what extent e-cigarette users noticed the European Union’s new legislation regarding e-cigarettes, and whether this may have influenced perceptions regarding addictiveness and toxicity. Data were obtained from yearly surveys (2015−2017) of the International Tobacco Control (ITC) Netherlands Survey. Descriptive statistics and Generalized Estimating Equations were applied. About a third of the e-cigarette users noticed the text warning (28%) and the leaflet (32%). When compared to tobacco-only smokers, e-cigarette users showed greater increases in perceptions regarding addictiveness (β = 0.457, p = 0.045 vs. β = 0.135, p < 0.001) and toxicity (β = 0.246, p = 0.055 vs. β = 0.071, p = 0.010). In conclusion, the new legislation’s noticeability should be increased.

Rationale: Efforts towards tobacco control are numerous, but relapse rates for smoking cessations remain high. Behavioral changes necessary for continuous cessation appear complex, variable and subject to social, biological, psychological and environmental determinants. Currently, most cessation studies concentrate on short-to midterm behavioral changes. Besides, they use fixed typologies, thereby failing to capture the temporal changes in smoking/cessation behaviors, and its determinants.Objective: To obtain long-term, data-driven longitudinal patterns or profiles of smoking, cessation, and related determinants in a cohort of adult smokers, and to investigate their dynamic links. Methods: The dataset originated from the International Tobacco Control (ITC) Netherlands Project, waves 2008 to 2016. Temporal dynamics of smoking/cessation, psychosocial constructs, and time-varying determinants of smoking were extracted with Group-Based Trajectory Modeling technique. Their associations were investigated via multiple regression models.Results: Substantial heterogeneity of smoking and cessation behaviors was unveiled. Most respondents were classified as persistent smokers, albeit with distinct levels of consumption. For a minority, cessation could be sustained between 1 and 8 years, while others showed relapsing or fluctuating smoking behavior. Links between smoking/cessation trajectories with those of psychosocial and sociodemographic variables were diverse. Notably, changes in two variables were aligned to behavioral changes towards cessation: decreasing number of smoking peers and attaining a higher self-perceived control.Conclusion: The unveiled heterogeneity of smoking behavior over time and the varied cross-dependencies between smoking data-driven typologies and those of underlying risk factors underscore the need of individually tailored approaches for motivational quitting.

For growth-rate retardation in commercial growing pigs suffering from non-infectious diseases, no biomarker is available for early detection and prevention of the condition or for the diagnosis of affected animals. The point in question is that the underlying pathological pathway of the condition is still unknown and multiple nutritional or management issues could be the cause of the disease. Common health status markers such as acute phase proteins, adenosine deaminase activity or total antioxidant capacity did not show any alteration in the saliva of animals with growth-rate retardation, so other pathways should be affected. The present study investigates saliva samples from animals with the same commercial crossbreed, sex and age, comparing control pigs and pigs with growth-rate retardation. A proteomics approach based on two-dimensional gel electrophoresis including mass spectrometry together with validation experiments was applied for the search of proteins that could help understand disease mechanisms and be used for early disease detection. Two proteins were detected as possible markers of growth-rate retardation, specifically S100A12 and carbonic anhydrase VI. A decrease in innate immune response was confirmed in pigs with growth-rate retardation, however further studies should be necessary to understand the role of the different CA VI proteoforms observed.

Introduction Since May 2020, the ban on production and sale of menthol and other flavored cigarettes will come into force in the European Union countries. Then millions of cigarette smokers can quit smoking or switch to other tobacco products, both conventional and novel. Study aim To assess the prevalence, socio-demographic characteristics, cessation behaviors and attitudes of menthol and other flavored cigarette smokers towards tobacco control policies and programs. Material and Methods Cross-sectional data were collected in 2016 among 10,760 adult current smokers from 8 European countries (EUREST-PLUS ITC Europe Project). Smokers of menthol, other flavoured, unflavored tobacco, or no usual brand were compared on smoking prevalence, socio-demographic characteristics, tobacco dependence, motivation to quit, self-efficacy, prior cessation behaviours, and attitudes towards tobacco control measures. The univariate data analysis adjusted to few confounding factors was made with the use of SPSS Complex Samples Package. Results Study results shows that around 7% of adults in all 8 analyzed European countries smoke menthol cigarettes and around 3% other flavored cigarettes. However, prevalence of menthol smoking substantially varies between countries (0.4% in Spain to 12.4% in England). Compared to other groups, menthol cigarette smokers are younger, more likely to be female, better educated, had higher household income, and smoked fewer cigarettes. In comparison to smokers of unflavored cigarettes, smokers of menthol and other flavoured cigarettes are less likely to smoke daily, tend to be less dependent and have higher quitting self-efficacy. In comparison to all other smokers, menthol smokers are less likely to support a ban on sale of flavored tobacco products, including menthol cigarettes. After enforcement of the ban, around a fifth of all menthol and flavored cigarette smokers intends to switch to another brand, and a third to reduce the amount they smoked or to quit smoking. Conclusions The EU TPD ban on sale of flavored cigarettes will affect one in ten smokers in the countries surveyed and provides an opportunity for targeting these groups with tobacco control policies and cessation programs. Lower tobacco dependence rates found in smokers of menthol and other flavored cigarettes may lead to greater success rates if quit attempts are strengthened by smoking cessation programs. However, smokers of different cigarette flavours in Europe differ on a range of smoking and cessation characteristics and may need different, target-tailored approaches.

The exoproteome of parasitic protists constitutes extracellular proteins that play a fundamental role in host-parasite interactions. Lytic factors, especially secreted proteases, are capable of modulating tissue invasion, thereby aggravating host susceptibility. Despite the important role of exoproteins during infection, the exoproteomic data on Histomonas meleagridis are non-existent. The present study employed traditional 1D-in-gel-zymography (1D-IGZ) and micro-LC-ESI-MS/MS (shotgun proteomics), to investigate H. meleagridis exoproteomes, obtained from a clonal virulent and an attenuated strain. Both strains were maintained as mono-eukaryotic monoxenic cultures with Escherichia coli. We demonstrated active in vitro secretion kinetics of proteases by both parasite strains, with a widespread proteolytic activity ranging from 17 kDa to 120 kDa. Based on protease inhibitor susceptibility assay, the majority of proteases present in both exoproteomes belonged to the family of cysteine proteases and showed stronger activity in the exoproteome of a virulent H. meleagridis. Shotgun proteomics, aided by customized database search, identified 176 proteins including actin, potential moonlighting glycolytic enzymes, lytic molecules such as pore-forming proteins (PFPs) and proteases like cathepsin-L like cysteine protease. To quantify the exoproteomic differences between the virulent and the attenuated H. meleagridis cultures, a sequential window acquisition of all theoretical spectra mass spectrometric (SWATH-MS) approach was applied. Surprisingly, results showed most of the exoproteomic differences to be of bacterial origin, especially targeting metabolism and locomotion. By deciphering such molecular signatures, novel insights into a complex in vitro protozoan- bacteria relationship were elucidated.

Introduction:There is clear evidence that the use of cessation aids significantly increases the likelihood of successful smoking cessation. The aim of this study was to examine quitting activity and use of cessation aids among smokers from various European countries. Subgroup differences were also examined for sex, income, education, and age in each country.Methods:Cross-sectional data were collected in 2016 from 10683 smokers in eight European countries participating in the ITC Project: England (n=3536), Germany (n=1003), Greece (n=1000), Hungary (n=1000), the Netherlands (n=1136), Poland (n=1006), Romania (n=1001), and Spain (n=1001). We measured quitting activity, including quit attempts in the previous 12 months and intention to quit, use of cessation aids (i.e. medication, quitlines, internet, local services, e-cigarettes), and whether respondents had received advice from health professionals about quitting and e-cigarettes.Results:Quit attempts were most common in England (46.3%) and least common in Hungary (10.4%). Quit intention was highest in England and lowest in Greece. Use of e-cigarettes to quit was highest in England (51.6%) and lowest in Spain (5.0%). Use of cessation aids was generally low across all countries; in particular this was true for quitlines, internetbased support, and local services. Receiving health professional advice to quit was highest in Romania (56.5%), and lowest in Poland (20.8%); few smokers received advice about e-cigarettes from health professionals. No clear differences were found for sex and income groups. Across countries, smokers with lower education reported less quitting activity.Conclusions:Quitting activity and use of cessation methods were low in most countries. Greater quit attempts and use of cessation aids were found in England, where large investments in tobacco control and smoking cessation have been made. Health professionals are important for motivating smokers to quit and promoting the effectiveness of various methods, but overall, few smokers get advice to quit.

Introduction: Little is known about the extent to which smoking restrictions are socially accepted in a country such as the Netherlands where smoking restrictions have been implemented and reversed several times. The current study assessed trends as well as factors associated with two indicators of social acceptance of smoking restrictions in the Netherlands: acceptance of smoking in public places and implementation of home smoking bans.Methods: We used data from the Dutch Continuous Survey of Smoking Habits (DCSSH) between 2005 and 2014 (n = 182 826). The DCSSH is a national population survey with a cross-sectional design in which respondents aged 15 years and older are surveyed weekly.Results: Acceptance of smoking in public places decreased for six out of eight included venues, with the largest decrease for smoking in restaurants. The decrease in acceptance was larger among younger respondents and smokers. Smoking on terraces was an exception: decrease in acceptance there was larger among older respondents and ex-smokers. Implementation of home smoking bans increased over time. Having implemented a home smoking ban was associated with being male, being younger, having a high socioeconomic status, and being ex-or never smoker.Conclusions: Social acceptance of smoking restrictions has increased in the Netherlands, despite a suboptimal implementation process of smoking restrictions. However, there is still potential for improvement as acceptance of smoking is still quite high for some public venues like bars. It is important to strengthen smoking restrictions in order to further denormalize smoking in the Netherlands.Implications: We examined the extent to which smoking restrictions are socially accepted in the Netherlands where smoking restrictions have been implemented and reversed several times. Acceptance of smoking in public places decreased and implementation of home smoking bans increased between 2005 and 2014. Social acceptance of smoking restrictions increased in the Netherlands despite a suboptimal implementation process of smoking restrictions. However, acceptance of smoking in bars remains relatively high.

Background: Attrition bias can affect the external validity of findings. This article analyses attrition bias and assesses the effectiveness of replenishment samples on demographic and smoking-related characteristics for the International Tobacco Control Netherlands Survey, a longitudinal survey among smokers. Methods: Attrition analyses were conducted for the first five survey waves (2008–12). We assessed, including and excluding replenishment samples, whether the demographic composition of the samples changed between the first and fifth waves. Replenishment samples were tailored to ensure the sample remained representative of the smoking population. We also constructed a multivariable survival model of attrition that included all five waves with replenishment samples. Results: Of the original 1820 respondents recruited in 2008, 46% participated again in 2012. Demographic differences between waves due to attrition were generally small and replenishment samples tended to minimize them further. The multivariable survival analysis revealed that only two of the 10 variables analysed were significant predictors of attrition: a weak effect for gender (men dropped out more often) and weak to moderate effects for age (respondents aged 15–24 years dropped out more than aged 25–39 years, who dropped out more than those aged 40+ years). Conclusions: Weak to moderate attrition effects were found for men and younger age groups. This information could be used to minimize respondent attrition. Our findings suggest that sampling weights and tailored replenishment samples can effectively compensate for attrition effects. This is already being done for the International Tobacco Control Netherlands Survey, including the categories that significantly predicted attrition in this study.

The growth promoting effect of supplementing animal feed with antibiotics like tetracycline has traditionally been attributed to their antibiotic character. However, more evidence has been accumulated on their direct anti-inflammatory effect during the last two decades. Here we used a pig model to explore the systemic molecular effect of feed supplementation with sub therapeutic levels of oxytetracycline (OTC) by analysis of serum proteome changes. Results showed that OTC promoted growth, coinciding with a significant down regulation of different serum proteins related to inflammation, oxidation and lipid metabolism, confirming the anti-inflammatory mechanism of OTC. Interestingly, apart from the classic acute phase reactants also down regulation was seen of a hibernation associated plasma protein (HP-27), which is to our knowledge the first description in pigs. Although the exact function in non-hibernators is unclear, down regulation of HP-27 could be consistent with increased appetite, which is possibly linked to the anti-inflammatory action of OTC. Given that pigs are good models for human medicine due to their genetic and physiologic resemblance, the present results might also be used for rational intervention in human diseases in which inflammation plays an important role such as obesity, type 2 diabetes and cardiovascular diseases.

BackgroundMuch attention has been directed towards the possible effects of e-cigarette advertisements on adolescent never smokers. However, e-cigarette advertising may also influence perceptions and behaviours of adult smokers. The aim of our study was to examine whether noticing e-cigarette advertisements is associated with current use of e-cigarettes, disapproval of smoking, quit smoking attempts, and quit smoking success.MethodsWe used longitudinal data from two survey waves of the ITC Netherlands Survey among smokers aged 16 years and older (n = 1198). Respondents were asked whether they noticed e-cigarettes being advertised on television, on the radio, and in newspapers or magazines in the previous 6 months.ResultsThere was a significant increase in noticing e-cigarette advertisements between 2013 (13.3%) and 2014 (36.0%), across all media. The largest increase was for television advertisements. There was also a substantial increase in current use of e-cigarettes (from 3.1% to 13.3%), but this was not related to noticing advertisements in traditional media (OR = 0.99, p = 0.937). Noticing advertisements was bivariately associated with more disapproval of smoking (Beta = 0.05, p = 0.019) and with a higher likelihood of attempting to quit smoking (OR = 1.37, p = 0.038), but these associations did not reach significance in multivariate analyses. There was no significant association between noticing advertisements and quit smoking success in either the bivariate or multivariate regression analysis (OR = 0.92, p = 0.807).ConclusionNoticing e-cigarette advertisements increased sharply in the Netherlands between 2013 and 2014 along with increased e-cigarette use, but the two appear unrelated. The advertisements did not seem to have adverse effects on disapproval of smoking and smoking cessation.

BackgroundMass media campaigns to encourage smoking cessation have been shown to be effective in a context of comprehensive tobacco control programme. The effectiveness of antismoking ads that evoke negative emotions remains unclear, in particular in countries with high smoking prevalence and among smokers with low perceived susceptibility, low self-efficacy or who are not users of smoking cessation services.ObjectiveTo evaluate short-term and long-term effects of a 1-month French national highly emotional media campaign, with a focus on these specific targets.DesignA 6-month longitudinal survey by Internet. A sample of 3000 smokers were interviewed before the media campaign (T0). They were contacted again just after (T1) and 6 months after the campaign (T2).OutcomesPerceived susceptibility to the risks of smoking, self-efficacy to quit smoking, use of smoking cessation services (quitline and website) and 7-day quitting.MethodsThe analysis was carried out on 2241 individuals who answered at T1 and T2. Multiple logistic regressions were computed to test the association between the change in each outcome at T1 and T2 and the level of exposure based on self-reported recall.ResultsSelf-reported recall was associated with an increase in perceived susceptibility and with use of cessation services. Campaign recall was also associated with higher 7-day quitting immediately after the campaign (OR=1.8 (1.0 to 3.2), PConclusionsFear-appeal mass media campaigns can be effective in encouraging cessation among smokers in a country with high smoking prevalence (France), but should be accompanied by convincing self-efficacy messages.

IntroductionThe aim of the study was to compare the construct validity and the predictive validity of three instruments to measure intention to quit smoking: a Stages of Change measure, the Motivation To Stop Scale (MTSS), and a Likert scale. We used the Theory of Planned Behavior as theoretical framework.MethodsWe used data from the International Tobacco Control Netherlands Survey. We included smokers who participated in three consecutive survey waves (n = 980). We measured attitude, subjective norm, and perceived behavioral control in 2012, intention to quit with three instruments in 2013, and having made a quit attempt in the last year in 2014. We conducted Structural Equation Modeling with three models for the instruments of intention separately and with one model that included the three instruments simultaneously.ResultsAll three instruments of intention were significantly and positively related to attitude and perceived behavioral control but none was related to subjective norm. All three instruments were significantly and positively related to making a quit attempt. The relation of the Likert scale with making a quit attempt (β = 0.38) was somewhat stronger than that of the Stages of Change measure (β = 0.35) and the MTSS (β = 0.22). When entering the three instruments together into one model, only the Likert scale was significantly related to making a quit attempt.ConclusionsAll three instruments showed reasonable construct validity and comparable predictive validity. Under the studied conditions, the Likert scale performed slightly better than the Stages of Change measure and the MTSS.ImplicationsAn assessment of the Stages of Change, the Motivation To Stop Scale, and a Likert scale showed comparable predictive and construct validity as measures for intention to quit smoking. All three instruments can be used in future research; however, under the studied theoretical framework, that is, the Theory of Planned Behavior, the Likert scale performed slightly better than the other two instruments.

The head kidney and spleen are major lymphoid organs of the teleost fish. The authors identify proteome profiles of head kidney and spleen of rainbow trout (Oncorhynchus mykiss) using a shotgun proteomic approach. Gene ontology annotation of proteins is predicted using bioinformatic tools. This study represents detailed proteome profiles of head kidney and spleen of rainbow trout, with a total of 3241 and 2542 proteins identified, respectively. It is found that lymphoid organs are equipped with a variety of functional proteins related to defense, receptor, signal transduction, antioxidant, cytoskeleton, transport, binding, and metabolic processes. The identified proteome profiles will serve as a template for understanding lymphoid organ functions in salmonids and will increase the amount of spectra information of rainbow trout proteins in the public data repository PRIDE. This data can be accessed via ProteomeXchange with identifiers PXD008473 and PXD008478.

Background Free or lower cost cessation medication is reported more often as a trigger to think about quitting by smokers with low education and/or income, but it is not clear whether this leads to actual quitting of smoking. The aim of this study was to investigate whether reporting free medication as trigger is related to medication use, reported quit attempts and quit success and whether this association is modified by education and income. Methods Data were derived from the 2013 and 2014 surveys of the International Tobacco Control (ITC) Netherlands (n=1164) and United Kingdom (n=768) cohort. From the 2013 survey, current smokers were selected who also participated in the 2014 follow-up survey. Logistic regression analyses were used to assess associations between free medication as a trigger to think about quitting in 2013 and the use of smoking cessation medication, quit attempts and smoking cessation in 2014. Results 37.0% of smokers in the UK and 24.9% of smokers in the Netherlands reported free or lower cost medication as trigger to think about quitting. Smokers who reported this trigger were more likely to have used cessation medication during a quit attempt both in the UK (OR = 4.19, p < 0.001) and in the Netherlands (OR = 2.14, p = 0.033). The association between free medication as trigger to think about quitting and quit attempts was significant in the UK (OR = 1.45, p = 0.030), but not in the Netherlands (OR = 1.10, p = 0.587). There was no significant association with quit success. Associations did not differ across income and education groups. Conclusions Free or lower cost smoking cessation medication may stimulate quit attempts and increase the use of cessation medication among smokers with low, moderate, and high socioeconomic status.

Background The Netherlands implemented smokefree legislation in July 2008. However, this legislation was reversed and reimplemented for small bars without employees several times. Since October 2014, the smokefree legislation applies to all bars again, but not all of them comply and designated smoking rooms are still allowed. Methods A nationally representative sample of adult smokers from the Netherlands was interviewed each year between 2008 and 2016 as part of the International Tobacco Control (ITC) Survey (n=1246-1773 per year). Trends in perceptions of rules about smoking in bars they visited, seeing people smoke in bars during the last visit, support for smokefree bars, and social acceptance of smoking in bars were examined with GEE analyses. Results There was a significant increase in smokers reporting that bars they visited had a total smoking ban (9% in 2008 to 45% in 2016), which was more often reported by older and high income smokers. Between 10 and 18% of smokers reported that they did not know the rules about smoking in bars; this was more often reported by older smokers. Reports of having seen people smoking in bars during the last visit decreased from 93% to 19%. Support for smokefree bars doubled (20% to 43%) and was higher among older and high educated smokers. Social acceptance of smoking in bars decreased (75% to 41%) and was higher among younger and low income smokers. Conclusions Eight years after the implementation of smokefree legislation in bars, smoking in bars has decreased but is far from eliminated in the Netherlands. Educational campaigns targeted at younger and lower socioeconomic status smokers are needed to inform these groups about the (need for) rules about smoking in bars. Now that only a minority of smokers think it is still acceptable to smoke in bars, the Dutch government should consider banning designated smoking rooms too.

Yersinia ruckeri is the causative agent of enteric redmouth disease in salmonids. Head kidney and spleen are major lymphoid organs of the teleost fish where antigen presentation and immune defense against microbes take place. We investigated proteome alteration in head kidney and spleen of the rainbow trout following Y. ruckeri strains infection. Organs were analyzed after 3, 9 and 28 days post exposure with a shotgun proteomic approach. GO annotation and protein-protein interaction were predicted using bioinformatic tools. Thirty four proteins from head kidney and 85 proteins from spleen were found to be differentially expressed in rainbow trout during the Y. ruckeri infection process. These included lysosomal, antioxidant, metalloproteinase, cytoskeleton, tetraspanin, cathepsin B and c-type lectin receptor proteins. The findings of this study regarding the immune response at the protein level offer new insight into the systemic response to Y. ruckeri infection in rainbow trout. This proteomic data facilitate a better understanding of host-pathogen interactions and response of fish against Y. ruckeri biotype 1 and 2 strains. Protein-protein interaction analysis predicts carbon metabolism, ribosome and phagosome pathways in spleen of infected fish, which might be useful in understanding biological processes and further studies in the direction of pathways.

Measures to discourage tobacco use can be effective in reducing smoking. In the Netherlands, some of these measures were changed or even temporarily or permanently reversed after implementation. This dissertation evaluates inconsistencies in the implementation process and the socioeconomic differences found. The study results show that inconsistent implementation of measures can affect the outcome. For instance, adjustments to the regulations regarding e-cigarettes are associated with changes in e-cigarette use. It is also shown that there are socioeconomic differences between the effects of some of these measures. Social acceptance of smoking in public places and at home is higher among smokers from low socioeconomic backgrounds. Low-income smokers of low education are more frequently encouraged to consider stopping by various measures to discourage tobacco use. In this dissertation, the validity of various instruments measuring intention to stop smoking is assessed. The three instruments that are studied are shown to be good predictors of attempts to stop smoking. They are also shown to be useful for future studies based on questionnaires.

Introduction: The aim of the current study is to investigate trends and socioeconomic differences in policy triggers for thinking about quitting in six European countries.Methods: Data were derived from all available survey waves of the International Tobacco Control (ITC) Europe Surveys (2003-2013). France conducted three survey waves (n = 1420-1735), Germany three waves (n = 515-1515), The Netherlands seven waves (n = 1420-1668), Ireland three waves (n = 582-1071), Scotland two waves (n = 461-507), and the rest of the United Kingdom conducted seven survey waves (n = 861-1737). Smokers were asked whether four different policies (cigarette price, smoking restrictions in public places, free or lower cost medication, and warning labels on cigarette packs) influenced them to think about quitting. Generalized Estimating Equation (GEE) models were estimated for each country.Results: Cigarette price was mentioned most often in all countries and across all waves as trigger for thinking about quitting. Mentioning cigarette price and warning labels increased after the implementation of price increases and warning labels in some countries, while mentioning smoking restrictions decreased after their implementation in four countries. All studied policy triggers were mentioned more often by smokers with low and/or moderate education and income than smokers with high education and income. The education and income differences did not change significantly over time for most policies and in most countries.Conclusions: Tobacco control policies work as a trigger to increase thoughts about quitting, particularly in smokers with low education and low income and therefore have the potential to reduce health inequalities in smoking. (C) 2015 Elsevier Ireland Ltd. All rights reserved.