The fungus Acremonium alternatum enhances salt stress tolerance by regulating host redox homeostasis and phytohormone signaling.

While endophytic fungi offer promising avenues for bolstering plant resilience against abiotic stressors, the molecular mechanisms behind this biofortification remain largely unknown. This study employed a multifaceted approach, combining plant physiology, proteomic, metabolomic, and targeted hormonal analyses to illuminate the early response of Brassica napus to Acremonium alternatum during the nascent stages of their interaction. Notably, under optimal growth conditions, the initial reaction to fungus was relatively subtle, with no visible alterations in plant phenotype and only minor impacts on the proteome and metabolome. Interestingly, the identified proteins associated with the Acremonium response included TUDOR 1, Annexin D4, and a plastidic K+ efflux antiporter, hinting at potential processes that could counter abiotic stressors, particularly salt stress. Subsequent experiments validated this hypothesis, showcasing significantly enhanced growth in Acremonium-inoculated plants under salt stress. Molecular analyses revealed a profound impact on the plant's proteome, with over 50% of salt stress response proteins remaining unaffected in inoculated plants. Acremonium modulated ribosomal proteins, increased abundance of photosynthetic proteins, enhanced ROS metabolism, accumulation of V-ATPase, altered abundances of various metabolic enzymes, and possibly promoted abscisic acid signaling. Subsequent analyses validated the accumulation of this hormone and its enhanced signaling. Collectively, these findings indicate that Acremonium promotes salt tolerance by orchestrating abscisic acid signaling, priming the plant's antioxidant system, as evidenced by the accumulation of ROS-scavenging metabolites and alterations in ROS metabolism, leading to lowered ROS levels and enhanced photosynthesis. Additionally, it modulates ion sequestration through V-ATPase accumulation, potentially contributing to the observed decrease in chloride content.

Adverse Effects of Meditation: Autonomic Nervous System Activation and Individual Nauseous Responses During Samadhi Meditation in the Czech Republic.

Buddhist meditation practices, including Samadhi meditation, which forms the basis for mindfulness practice, are broadly promoted as pathways to wellbeing, but evidence of their adverse effects is emerging. In a single-group observational study with assessments of autonomic system before, during, and after Samadhi meditation, we explore the relationship between post-meditation nausea symptoms and the degree of change in autonomic system activity during meditation as compared to before and after in 57 university students (42 women; mean age = 22.6) without any previous experience in meditation or yoga practices. We hypothesize that nauseous feelings in meditation are connected to a rapid increase of activity in the sympathetic nervous system, as indicated by decreased heart-rate variability (HRV). We additionally explore links between meditation-induced nausea and two markers of parasympathetic activity: increased HRV and vasovagal syncope. Engaging in meditation and increased nausea during meditation were both associated with increased markers of HRV parasympathetic activity, but 12 individuals with markedly higher nausea demonstrated increased HRV markers of sympathetic activity during meditation. Vasovagal syncope was observed but found to be unrelated to nausea levels. Drivers of adverse effects of meditation in some individuals require further investigation.

Increasing prevalence of mental disorders in smokers seeking treatment of tobacco dependence: a retrospective observational study.

BACKGROUND: There has been a noticeable relative increase in psychiatric comorbidities among smokers as opposed to the general population. This is likely due to comparatively slower decrease in smoking prevalence among individuals with mental health conditions. This study aims to assess the prevalence trend of past or current mental health disorders in individuals seeking specialized smoking cessation assistance. METHODS: We conducted a retrospective single-centre observational study to assess the presence of mental disorders such as anxiety, depression, bipolar affective disorder, or schizophrenia in personal history of 6,546 smokers who sought treatment at the Centre for Treatment of Tobacco Dependence in Prague, Czech Republic between 2006 and 2019. The study examined the impact of gender, age, and the effect of successive years on the prevalence of the mental disorders using Poisson distribution regression. RESULTS: In the studied cohort, 1,743 patients (26.6%) reported having one or more mental disorders. Compared to patients without a psychiatric disorder, they exhibited similar levels of carbon monoxide in expired air (mean 17 ppm, SD 11 ppm) and scored one point higher on the Fagerström Test of Cigarette Dependence. Among smokers with a mental disorder, women were more prevalent (62%) than men (38%). The prevalence of mental disorders increased on average by 4% every year, rising from 23% in 2006 to 35% in 2019. CONCLUSIONS: Consistent with the observation that the prevalence of smoking among people with any mental disorder is higher and declining at a slower rate than in the general population, there is a steadily increasing percentage of these patients seeking specialized treatment over time. Professionals who offer tobacco dependence treatment should be aware of the upward trend in psychiatric disorders among smokers, as more intensive treatment may be needed. Similarly, psychiatric care should pay attention to smoking of their patients.

Minnesota Tobacco Withdrawal Scale revision 2020, validated Czech translation.

Most smokers are addicted to tobacco or nicotine not only psychosocially but also physically, what results in withdrawal symptoms after they miss their dose. These are one of the major barriers to quitting. In the treatment of tobacco dependence, the evaluation of the degree of withdrawal symptoms helps to guide psychobehavioral intervention and, above all, the choice of the type, dose, and duration of pharmacotherapy. The Minnesota Withdrawal Symptoms Scale is one of the most widely used for this purpose, revised in 2020. We present a validated translation of this update.

Relationship of Weight Concerns to Body Mass Index in a Tobacco Treatment Program.

Objectives: Our aim was to examine the relationship between weight concerns (WC) and smoking cessation. Methods: WC was assessed before smoking cessation treatment in 671 adult patients who completed a 12-month follow-up visit at the Centre for Tobacco-Dependent, Prague, Czech Republic, from 2013 through 2019. We evaluated the abstinence rate at 12-month follow-up. Results: Among 669 patients with baseline WC (mean age, 43.4 years), 47% were women (145/306) and 21% were men (78/363). No association existed between WC and abstinence at 12 months. Smokers with obesity had increased fear of gaining weight (34% versus 24% of overweight and 23% of healthy-weight smokers) (p=.034) and were less confident in their ability to maintain their current weight (36% versus 55% of overweight smokers and 59% of healthy- weight smokers) (p<.001). Conclusions: Many smokers are concerned about gaining weight after stopping smoking, but in this cohort of patients, having WC was not associated with 12-month abstinence, but obesity or overweight was related to being afraid of postcessation weight-gain and low confidence for maintaining weight. Practitioners should be aware of the prevalence of WC in those stopping smoking and should address concerns, such as poor motivation and low confidence about managing their weight.

Colour Categorization and its Effect on Perception: A Conceptual Replication.

The presented study examines the question of colour categorization in relation to the hypothesis of linguistic relativity. The study is based on research conducted by Gilbert et al. (2006) and their claim that linguistic colour categorization in a particular language helps colour recognition and speeds the process of colour discrimination for colours from different linguistic categories but only for the right visual field. Our study approached the research question differently. We used the same methodology as Gilbert's team et al. (2006), but we used different colour categories in the Czech language and significantly enlarged the number of participants to 106 undergraduate psychology students. Our results show that the fastest reaction times were in trials when the target was located in the left visual field, quite opposite from the Gilbert's et al. (2006) study. We argue that this finding is based on different processes than simple colour linguistic categorisation and attentional processes actually play an important role in the task.

Tobacco dependence treatment guidelines.

These recommendations are relevant for all clinical fields and all medical professions, although for some more and for some less. Smoking is the cause of every sixth death in the Czech Republic and is related to diseases of all organ systems. Effective treatment for the diagnosis of F17 exists and should be offered and available to approximately 2 million smokers in the Czech Republic as a standard part of medical and preventive care according to time availability - from brief intervention in frame of each clinical contact with the patient to intensive care. It includes psycho-socio-behavioral support and pharmacotherapy. It should cover all professions in clinical medicine - according to WHO recommendations, especially doctors, nurses, pharmacists and dentists and should be reimbursed within health systems. It should also be offered in other services like e.g., social or addictology care. From an economic point of view, it is one of the most cost-effective interventions in medicine.

SNPs within CHRNA5-A3-B4 and CYP2A6/B6, nicotine metabolite concentrations and nicotine dependence treatment success in smokers.

AIM: Plasma values of nicotine and its metabolites are highly variable, and this variability has a strong genetic influence. In our study, we analysed the impact of common polymorphisms associated with smoking on the plasma values of nicotine, nicotine metabolites and their ratios and investigated the potential effect of these polymorphisms and nicotine metabolite ratios on the successful treatment of tobacco dependence. METHODS: Five variants (rs16969968, rs6474412, rs578776, rs4105144 and rs3733829) were genotyped in a group of highly dependent adult smokers (n=103). All smokers underwent intensive treatment for tobacco dependence; 33 smokers were still abstinent at the 12-month follow-up. RESULTS: The rs4105144 (CYP2A6, P<0.005) and rs3733829 (EGLN2, P<0.05) variants were significantly associated with plasma concentrations of 3OH-cotinine and with 3OH-cotinine: cotinine ratios. Similarly, the unweighted gene score was a significant (P<0.05) predictor of both cotinine:nicotine and 3OH-cotinine:cotinine ratios. No associations between the analysed polymorphisms or nicotine metabolite ratios and nicotine abstinence rate were observed. CONCLUSION: Although CYP2A6 and EGLN2 polymorphisms were associated with nicotine metabolism ratios, neither these polymorphisms nor the ratios were associated with abstinence rates.

Comparison between success rates for smokers re-treated by a smokers' clinic and success rates for smokers treated for the first time.

AIMS: To compare success rates and characteristics of smokers treated a second time by a smokers' clinic with success rates of their first treatment. DESIGN: Retrospective cohort study. SETTING: Tobacco Dependence Treatment clinic in Prague, Czech Republic, between 2005 and 2017. PARTICIPANTS: A total of 5225 smokers treated either once (n = 5006, single treatment sample, SS) or also second time (n = 219, re-treated sample, RS), on average 4.47 years after the first visit. INTERVENTION: Smokers received intensive treatment of tobacco dependence with pharmacotherapy options. Outcomes were evaluated after 1 year. In case of failure or relapse, participants could undergo re-treatment in the same setting at least 1 year after the start of the first treatment. MEASUREMENTS: Twelve-month self-reported continuous abstinence; CO-validated (≤ 6 parts per million); number of visits; type of pharmacotherapy; mental health history; Fagerström Test for Cigarette Dependence; time between first and second treatment. RESULTS: The abstinence rate in the SS was 34.8% [95% confidence interval (CI) = 33.4%, 36.1%] and in the RS was 37% (95% CI = 30.6%, 43.8%) and 39.7% (95% CI = 33.2%, 45.5%) for their first and second treatments, respectively. The samples were comparable on smoking and socio-demographic characteristics and pharmacotherapy used, but the RS in the second treatment had a higher prevalence of diagnosed mental health disorder at 39.3% (95% CI = 32.8%; 46.1%) compared with 23.7% (95% CI = 22.5%; 24.9%) in the SS. Participants who initiated their second quit attempt 1 to 2 years after the first one were less successful than those who initiated their second quit attempt later (25 versus 43%; P < 0.05). The results of the first treatment cycle were not found to be a reliable predictor for outcomes of the second cycle of treatment in univariate or multivariate logistic regression (odds ratio = 1.35, 95% CI = 0.70-2.63, P = 0.373). CONCLUSION: In Prague, Czech Republic, smokers re-attending stop-smoking treatment more than 2 years after their previous quit attempt appear to achieve similar success rates to those being treated for the first time.

Tobacco dependence treatment in the Czech Republic: history, presence, and future.

The tobacco dependence treatment has a relative short history. It has become available in various intensity within our healthcare in the last more than two decades. The Society for the Treatment of Tobacco Dependence in cooperation with the Czech Medical Chamber initiated the establishment of Tobacco Dependence Treatment Centers, managing their database (N = 43) on its website (www.slzt.cz), as well as a directory of outpatient doctors providing this treatment (N = 214), and consulting pharmacies (N ~ 150). On this website, there are also mobile apps for quitting smoking in Czech, the national quit line, addictology clinics dealing with tobacco dependence (N = 3), a link to published treatment recommendations and other information. All doctors, nurses and other healthcare professionals routinely should apply brief intervention lasting at least tens of seconds, and also other diverse forms of assistance should be available from leaflets to intensive treatment. We also focus on the necessary future direction of this field of medicine.

Czech and Slovak life scripts: the rare case of two countries that used to be one.

In this study, we examined cultural life scripts in two countries, the Czech Republic and Slovakia. The cultural life script is semantic knowledge about culturally shared expectations regarding the order and timing of important life events during an idealised life course. For many decades, Slovakia and the Czech Republic were one country: Czechoslovakia. After a regime change in 1989 and the separation of the Czech Republic and Slovakia in 1993, their two cultures have been evolving and changing independently from one another, making these countries interesting for examining life scripts. We found that the cultural life scripts provided by Slovak and Czech participants shared 25 event categories, representing 89.3% of event categories in the Czech sample and 80.6% of event categories in the Slovak sample (including the category Other). However, participants also reported unique event categories to each culture (10.7% of unique event categories in the Czech sample and 19.4% in the Slovak sample), reflecting the specific cultural characteristics of these two separate countries. Reported events were listed in the same order they are expected to happen during the life span, were mostly positive, and showed a lifespan distribution consistent with the reminiscence bump. Participants showed higher agreement in the age estimates of positive events, compared to neutral and negative ones. Events were mostly social, in contrast to biological events. All these findings are consistent with the life script literature.

Hospital as a smoke-free workplace.

OBJECTIVES: The primary aim of a hospital is to promote/improve and restore health, thus smoking, whether in its passive or active form, should be banned in all hospital premises for the benefit of employees and patients alike. The Global Network for Tobacco Free Healthcare Services (GNTH) is an international non-profit association formed in 1999. The GNTH's mission is to implement tobacco-free policies to create a healthy workplace and patients' environment; help physicians, nurses, and other healthcare workers to stop smoking; and educate all caregivers about tobacco dependence treatment and support them in providing smoking cessation interventions. METHODS: Implementation standards and a system of their self-audit for all participating hospitals were developed by the GNTH. We describe both the international and Czech networks, recommended methods for programme implementation and results of self-audit questionnaires completed by Czech participating hospitals. RESULTS: Worldwide, there are 19 national networks with 1,672 members including 56 gold forum members. To date, the largest network has been formed in France (670 members), followed by Spain (580) and Taiwan (209). After the first Czech institution (Prague-based General University Hospital) joined GNTH in 2010, the Czech Republic established its national network in 2017 currently comprising 10 members, of this number 1 gold, 3 silver and 6 bronze national certification level members. The main barriers to better outcomes in the Czech Republic include smoking on outdoor hospital grounds, lack of pharmacotherapy reimbursement and time, and inadequate staff education in the field of tobacco dependence treatment. CONCLUSIONS: The Global Network's mission is to advocate, recruit and enable healthcare services and professionals to implement and sustain effective tobacco management and cessation policies in accordance with the WHO Framework Convention on Tobacco Control (FCTC). A systematic approach supports the quality of care and treatment outcomes for patients as well as healthy workplace conditions for the staff.

Early weight gain after stopping smoking: a predictor of overall large weight gain? A single-site retrospective cohort study.

OBJECTIVES: Most people gain weight on stopping smoking but the extent of weight gain varies greatly. Interventions aimed at all quitters to prevent weight gain on cessation have proven unpopular but targeting people who have gained excess weight immediately after quitting may improve uptake and cost-effectiveness. We examined whether early large postcessation weight gain predicts overall large weight gain. DESIGN: Retrospective cohort study. SETTING: Primary care setting-smoking cessation centre in Prague, Czech Republic. PARTICIPANTS: Out of 3537 patients treated between 2005 and 2013, 1050 were continuous abstainers (verified by carbon monoxide measurement) at 1-year follow-up and formed the cohort of the current report. 48.7% were women (n=511) with the mean age of 46 (±14.4) years. METHODS: In this retrospective cohort study, all patients underwent usual tobacco dependence treatment using evidence-based methods. Weight was measured prior to smoking cessation and at each visit after quitting. RESULTS: The mean weight gain in the first month (n=763) was 0.79% (±2.03%), in the second month (n=646) was 1.49% (±2.58%), for the third month (n=566) 2.33% (±3.44%) and 4.1% (±5.31%) after 1-year follow-up (n=1050). The regression coefficient per 1% rise in the first 3 months was +0.13% (95% CI -0.04% to 0.30%). A receiver operating curve analysis showed that patients gaining more than 0.98% of their baseline weight during first 3 months had a sensitivity of 66% and specificity of 44% for gaining 7% or more weight by 12 months. In addition, lower body mass index and an increase in appetite at 3 months after quitting were associated with greater weight gain, while using nicotine replacement therapy was associated with less weight gain at 1-year follow-up. CONCLUSIONS: People who stop smoking and gain a larger amount of weight early after quitting are not more likely to gain excessively at 1 year.

Fecal zonulin is elevated in Crohn's disease and in cigarette smokers.

OBJECTIVES: Human zonulin is a protein that increases permeability in the epithelial layer of the small intestine by reversibly modulating the intercellular tight junctions. There is not sufficient information available about zonulin's participation in inflammatory bowel diseases (IBD). The aim of this study was therefore to investigate fecal and serum zonulin in IBD patients and its relation to the disease localization, behavior and smoking status. DESIGN AND METHODS: Forty IBD patients and forty healthy persons were examined for fecal and serum zonulin concentrations by competitive ELISA (DRG International Inc). Values were correlated to IBD type, localization and behavior, and smoking. RESULTS: Serum and fecal zonulin were significantly higher in patients with Crohn's disease compared to ulcerative colitis (p = 0.038 for fecal zonulin, and p = 0.041 for serum zonulin concentrations). No association of serum or fecal zonulin was found with respect to IBD localization and behavior. The only difference was found with respect to smoking. Both the IBD cohort and healthy smokers showed significantly higher fecal zonulin levels (median 203 ng/mL) compared to non-smokers (median 35.8 ng/mL), p < 0.001. CONCLUSIONS: Fecal and serum zonulin levels are elevated in patients with active Crohn's disease but not with ulcerative colitis. High fecal zonulin levels in smokers irrespective of IBD point to the significant and undesirable up-regulation of gut permeability in cigarette smokers.

[What does "Smoke-free Hospital" mean?] / Co znamená "Nekurácká nemocnice"?

"Smoke-free Hospital" is the name of a gradual and long-term project, with the really smoke-free hospital as the final goal. The project is based on the process itself that leads to such a status. European Network of Smoke-free Hospitals (ENSH, www.ensh.org) provides detailed instructions and recommendations regarding actions to be implemented. The main objective of the network is "to deliver safe quality care in relation to tobacco for every service user, every time and everywhere".Each hospital could identify its own sub-goals and methods of implementation, but it is important to assess the achieved steps regularly. It is advisable to focus attention e.g. to introduction of non-smoking areas, monitoring tobacco use among both patients and staff, providing brief interventions in smoking cessation as well as intensive tobacco dependence treatment, training of staff in intervening, organizing health promoting events both for the staff and the public. Several actions in 2016 connected with ENSH in the Czech Republic and on the international level are listed. The new Czech legislation regarding smoking in hospitals is also mentioned.

Weight Concerns Associated With Delay in Quit Date But Not Treatment Outcomes: A Czech Republic Experience.

BACKGROUND: Weight concerns are prevalent in smokers and may reduce the success rate of quitting. This concept has been primarily studied on US populations and it is unknown how weight concerns may differ cross-culturally. This study examined the role of weight concern in European smokers wishing to stop smoking. METHODS: A sample of 593 smokers (299 men and 294 women, mean age 38 years) utilizing the Centre for Tobacco-Dependent in Prague, Czech Republic, between 2010 and 2013 were studied. Weight concerns were assessed at baseline prior to treatment by evidence-based stop smoking methods. Abstinence was evaluated at 12 months post baseline. RESULTS: Approximately 34% of all patients (204/593) were classified as weight concerned (by indicating on the Weight Concern Scale that they would return to smoking after any weight gain) at the time they sought treatment. Among all men, 19.4% (58/299) were weight concerned and among all women, 49.7% (146/294) were weight concerned. Among females, weight-concerned smokers were of similar weight, but younger (p < .001), and had been smoking cigarettes for fewer years (p = .002) compared with those without weight concerns, whereas the male weight-concerned smokers were significantly (p = .030) heavier than those without weight concerns. Although the presence of weight concern was associated with a delay in setting a quit date (log-rank test p = .019), it was not associated with abstinence at one year. CONCLUSION: The quit success rate of weight-concerned smokers in Czech Republic did not differ from those without weight concern when utilizing an individualized smoking cessation treatment program. Individually tailored tobacco dependence treatment could help to prevent weight concern from affecting successful quitting. IMPLICATIONS: This study adds the new cross-cultural aspect of post-cessation weight concern. Weight concern has been studied primarily on US populations and our sample consists of European sample of smokers. Additionally, we have found that the presence of weight concern lead to delay in setting a quit date, but the success rate of those weight concerned did not differ from those without weight concern. Thus, it is possible, that this individualized evidence-based tobacco treatment program was able to prevent weight concern impact towards successful quitting.

Depression and Smoking Cessation: Evidence from a Smoking Cessation Clinic with 1-Year Follow-Up.

BACKGROUND: Smoking is more prevalent among people with depression. Depression may make cessation more difficult and cessation may affect depression symptoms. PURPOSE: The aims of this study were to assess the associations between (1) baseline depression and 1-year smoking abstinence and (2) abstinence and change in depression. METHODS: Observational study using data collected routinely in a smoking cessation clinic in the Czech Republic from 2008 to 2014. Aim 1: N = 3775 patients; 14.3% reported mild and 15.4% moderate/severe baseline depression levels measured using Beck's Depression Inventory (BDI-II). Logistic regressions assessed if depression level predicted 1-year biochemically verified abstinence while adjusting for patient and treatment characteristics. Aim 2: N = 835 patients abstinent at 1 year; change in depression was analysed using Chi-square statistics, t test and mixed method analyses of variance. RESULTS: Rate of abstinence was lower for patients with mild (32.5%, OR = 0.68; 95% CI: 0.54 to 0.87, p = 0.002) and moderate/severe depression (25.8%; OR = 0.57, 95% CI: 0.45 to 0.74, p < 0.001) compared with patients without depression (40.5%). Across abstinent patients, the majority with baseline depression reported lower depression levels at follow-up. Overall mean (SD) BDI-II scores improved from 9.2 (8.6) to 5.3 (6.1); t(834) = 14.6, p < 0.001. There were significant main effects of time (F(1832) = 880.8, p < 0.001, partial η2 = 0.51) and baseline depression level (F(2832) = 666.4, p < 0.001, partial η2 = 0.62) on follow-up depression and a significant depression * time interaction (F(2832) = 296.5, p < 0.001, partial η2 = 0.42). CONCLUSIONS: In this effective smoking cessation clinic, depression at the start of treatment predicted reduced smoking abstinence 1 year later. Patients abstinent from smoking experienced considerable improvement in depression.

[Tobacco dependence treatment guidelines]. / Doporucení pro lécbu závislosti na tabáku.

Tobacco dependence causes every sixth death in the Czech Republic and is associated with diseases of the whole body. Treatment of tobacco dependence should be a standard part of clinical care to the extent of time available--from brief intervention at each clinical contact with patients up to intensive treatment. It includes psycho-socio-behavioural support and pharmacotherapy. It should apply to all professions in clinical medicine--as recommended by WHO mainly doctors, nurses, pharmacists and dentists, and should be covered within health care systems. Economically, it is one of the most cost-effective interventions in medicine. In our population over 15 years about 30% smokes (about 2.2 million people), diagnosis F17 relates to approximately 1.75 million people (about 80% of smokers).

[Centres for Tobacco-Dependent in the Czech Republic in 2012: overview and economy]. / Centra pro závislé na tabáku v Ceské republice v roce 2012: prehled, ekonomika.

BACKGROUND: Since 2005, in the Czech Republic the Centres for Tobacco-Dependent are being established at hospitals. METHODS: Evaluation of the activity of these 37 centres in 2012, economic analysis of treatment costs and assessment of the cost of life year gained (LYG). RESULTS: Most of the centres (26 of 37) are based at pulmonary clinics with opening hours for smokers: on average 7 hours/week. Treatment codes 25501 and 25503 are used at 28 centres. Entry visit usually takes on average 61 minutes, follow-up visits 22 minutes. Nicotine replacement therapy and varenicline are indicated in all centres, but only 14 centres use bupropion. Virtually all centres use links to other clinical disciplines, about 10.5 % of patients are sent to other departments. The most common barriers for wider activity are insufficient salaries and staffing. In 2012, the Centre for Tobacco-Dependent at the 3rd Medical Department, 1st Faculty of Medicine, Charles University in Prague and the General University Hospital treated 430 patients for the price of 3792 CZK per treated patient ( 150 Euro), respectively, with 38 % success rate for the price of 10,003 CZK per abstinent patient ( 400 Euro), or for 1,334 CZK per LYG ( 50 Euro/LYG). CONCLUSION: In the future it would be good to improve working conditions in centres and to take advantage of their potential for the indispensable, effective and highly cost-effective treatment.