The Italian National Institute of Health helpline to quit tobacco and nicotine dependence: 20 years of activity.

INTRODUCTION: Tobacco use is one of the world's leading preventable causes of death and is a major preventable risk factor of non-communicable diseases. Although smokers are aware of the health risks, their attempts to quit often fail, primarily due to the strong nicotine and/or tobacco dependence. Antismoking helplines have become an integral part of tobacco control efforts in many countries. In Italy, the ISS Antismoking Helpine is active since 2000. MATERIAL AND METHODS: The professional staff of the ISS Antismoking Helpline have gathered socio-demographic and smoking-related data via an electronic form, related to the received calls. The collected data have been processed in a dedicated database and analyzed to monitor the use and the quality of the service. In this study, a descriptive statistical analysis was conducted to inform about the activity of the helpline over the years. RESULTS: From May 2003 to June 2023 the helpline received 99,423 calls. Most smokers called to receive "support to quit" (82.6%). Counselling was provided in 11.4% of cases, and in the last two years has been strongly increased (40.0% of cases). The percentage of users requesting information on emerging tobacco and nicotine products is 1.2%, even if in 2023 this percentage has risen significantly (6.0%). Two legislative measures (in 2012 and in 2016) required to add the helpline number to all packets of tobacco cigarettes. Accordingly, the offer of counselling increased from 2.6% to 12.2%. CONCLUSIONS: The available resources in tobacco control, including the helpline, are still not sufficient to meet all the users needs. Adequate policies and stable funding to fight tobacco and nicotine dependence need increased commitment from government institutions to ensure equal access to treatments for all Italian citizens.

ViolHelp: results of a pilot study to identify potential warning signs and risk factors for self- and hetero-directed violence in the calls received by the Helplines of the Italian National Institute of Health.

BACKGROUND: Self- and hetero-directed violence (SHDV) is a serious public health problem and a complex phenomenon, influenced by individual and environmental factors. SHDV may occur particularly in moments of personal, economic and/or social crisis. During the COVID-19 pandemic, the ISS-Helplines operators have perceived an increase in psychological distress and self-isolation among callers. The ViolHelp project aimed at identifying potential warning signs and risk factors of SHDV emerging in the activity of the ISS-Helplines (Istituto Superiore di Sanità, ISS, Italian National Institute of Health). MATERIALS AND METHODS: A dashboard collecting warning signs and risk factors of SHDV was developed to be used during the ISS-Helplines activity. RESULTS: In one year of data collection, 135 calls were compiled. In 106 calls, callers referred experienced violence: 72 self-directed violence (SDV), 20 hetero-directed violence (HDV), 14 both. The most frequent warning signs and risk factors for SDV were desire to die (68.6%), previous suicide attempts (31.4%) and threat of self-harm (25.6%); for HDV were depressed mood (32.4%), diagnosis of pathology and/or psychiatric disorders, desire to die, use of psychotropic drugs, and alcohol abuse (29.4%). CONCLUSIONS: The results of this pilot project show the importance of being able to read the warning signs and to create a network that can improve information, prevention and support activities for people at risk of violence and their families.

Psychosocial risk and protective factors for youth problem behavior are associated with food addiction in the Generation Z.

Objective: Food Addiction (FA) and other well-known risk behavior as substance misuse tend to co-occur and may share similar risk and protective factors. The aim of this study was to assess the association between the diagnosis/severity of FA and psychosocial domains typically related to risk behavior syndrome in a large, nationally representative community sample of Generation Z underage Italian students. Method: The sample consisted of 8,755 students (3,623 from middle schools, 5,132 from high schools). A short version of the Yale Food Addiction Scale 2.0 was administered to evaluate FA. Risk and protective factors related to demographic, personality, behavior, and family variables were examined. Stepwise multivariate logistic and linear regressions were conducted. Results: The prevalence of FA was 30.8%. Female gender, social anxiety and depression symptoms, social withdrawal risk, Internet gaming disorder, social media addiction, current substance use, social challenge engagement and experienced doxing boosted the chance of FA diagnosis, whereas eating fruit and vegetables, playing competitive sports and an average sleep duration of 7-8 h per night reduced these odds. FA severity was significantly and positively associated with trait impulsiveness, social anxiety and depressive symptoms, risk of social withdrawal, recent substance use, social media, and gaming addiction, doxing suffered and risky social challenges participation. Negative associations between the severity of FA and fruit and vegetable diet habits were found. Conclusion: Our findings confirm that FA is widespread among Italian adolescents. The associations between the diagnosis and severity of FA and psychosocial risk factors for health, including, addictive and deviant behaviors related to digital misuse, suggest its belonging to the risk behavior constellation. Health promotion schemes based on a multicomponent strategy of intervention should consider the inclusion of FA and its psychosocial correlates.

Smoking intensity changes during the COVID-19 pandemic waves in a cohort of smokers in Italy.

INTRODUCTION: COVID-19 lockdown in Italy resulted in increased smoking consumption, mainly associated with mental distress. This study aims to update previous findings investigating changes in smoking intensity during the whole COVID-19 pandemic. METHODS: This analysis was carried out within the "LOST IN ITALY" ("LOckdown and lifeSTyle IN ITALY") and "LOST IN TOSCANA" studies on 880 smokers with information collected during main pandemic peaks. Changes in cigarettes/day were investigated in association with survey-periods, socio-demographic and psychological characteristics through a linear mixed-model. RESULTS: Net of psychological distress and socio-demographic variables, in comparison to pre-pandemic period cigarettes/day increased by 1.16 during lockdown, and remained over half higher subsequently. In the overall period, an increase of >1 cigarette/day was also associated to lower education, older age, male gender and psychotropics drugs use. CONCLUSIONS: After 2 years of pandemic, cigarettes/day have not yet returned to the pre-pandemic levels, mainly due to socio-demographic factors, but also to nicotine addiction, that tends to stabilize consumption.

An Item Response Theory-Based Scoring of the South Oaks Gambling Screen-Revised Adolescents.

The South Oaks Gambling Screen-Revised Adolescent (SOGS-RA) is one of the most widely used screening tools for problem gambling among adolescents. In this study, item response theory was used for computing measures of problem gambling severity that took into account how much information the endorsed items provided about the presence of problem gambling. A zero-inflated mixture two-parameter logistic model was estimated on the responses of 4,404 adolescents to the South Oaks Gambling Screen-Revised Adolescent to compute the difficulty and discrimination of each item, and the problem gambling severity level (θ score) of each respondent. Receiver operating characteristic curve analysis was used to identify the cutoff on the θ scores that best distinguished daily and nondaily gamblers. This cutoff outperformed the common cutoff defined on the sum scores in identifying daily gamblers but fell behind it in identifying nondaily gamblers. When screening adolescents to be subjected to further investigations, the cutoff on the θ scores must be preferred to that on the sum scores.

The impact of COVID-19 lockdown on gambling habit: A cross-sectional study from Italy.

BACKGROUND AND AIMS: Few preliminary studies have shown an impact of COVID-19 confinement on gambling habits. We aim to evaluate short-term effects of lockdown restrictions on gambling behaviors in Italy. METHODS: Within the project Lost in Italy, a web-based cross-sectional study was conducted on a representative sample of 6,003 Italians aged 18-74 years, enrolled during April 27-May 3 2020, and were asked to report gambling activity before the lockdown and at the time of interview. RESULTS: The prevalence of participants reporting any gambling decreased from 16.3% before lockdown to 9.7% during lockdown. Traditional gambling decreased from 9.9 to 2.4% and online gambling from 9.9 to 8.0%. Among gamblers, median time of gambling grew from 4.5 to 5.1 h/month. Among non-players before lockdown, 1.1% started playing. Among players before lockdown, 19.7% increased gambling activity. Multivariate analysis showed an increase in gambling activity in younger generations (p for trend = 0.001), current smokers (odds ratio, OR 1.48), users of electronic cigarettes (OR 1.63), heated tobacco products (OR 1.82), cannabis (OR 5.16), psychotropic drugs (OR 3.93), and subjects having hazardous alcohol drinking (OR 1.93). Self-reported low quality of life (OR 1.97), low sleep quantity (OR 2.00), depressive symptoms (OR 3.06) and anxiety symptoms (OR 2.93) were significantly related to an increase in total gambling activity during lockdown. DISCUSSION AND CONCLUSIONS: Although gambling substantially decreased during lockdown, time spent in gambling slightly increased. The strong relationship found between compromised mental health and addictive behaviors calls for urgent policies to prevent vulnerable populations from increasing and developing severe gambling addiction.

Stimulant substance use and gambling behaviour in adolescents. Gambling and stimulant use. / Uso de sustancias estimulantes y comportamiento de juego en adolescentes. Juego y uso de estimulantes.

Gambling is widely recognized as an important public health problem. Despite the rising use of stimulant substances among adolescents, there are still very few studies focusing on whether adolescents' use of stimulants is associated with their gambling behaviour. Therefore, the aim of this study was to investigate the association between gambling habits and consumption of stimulants such as coffee, energy drinks, and new psychoactive substances in a sample of Italian adolescents. A survey was conducted in 2017 with a representative sample of Italians between the ages of 14-17 years, comprising 15,833 students attending 201 secondary schools. Logistic regression analyses were run to assess the association between at-risk/problem gambling (O1) and independent predictors: the model included independent variables (coffee, energy drinks and new psychoactive substance consumption) and covariates (demographic variables, social environment variables and risk-taking behaviour variables). A sensitivity analysis was also conducted to examine a second dependent variable regarding any experience of gambling behaviour (O2). Adolescents who were at-risk gamblers or problem gamblers were significantly more likely to consume energy drinks than non-gamblers or not-at-risk gamblers. A similar pattern was seen for consumption of new psychoactive substances. No significant association emerged with coffee consumption. The sensitivity analysis showed that, compared with non gamblers, the group of gamblers had higher odds for frequent coffee consumption, as well as for consumption of energy drinks and/or new psychoactive substances. Screening for gambling and stimulant use may provide important information, as it may be necessary to take action to reduce stimulant substance use as part of efforts to deal with unhealthy gambling habits.

El juego es un importante problema de salud pública ampliamente reconocido. A pesar del creciente uso de sustancias estimulantes entre los adolescentes, todavía son escasos los estudios centrados en verificar la existencia de una asociación entre el uso de estimulantes y los comportamientos relacionados con el juego en adolescentes. Por tanto, este estudio tuvo como objetivo investigar la asociación entre los hábitos relacionados con el juego y el consumo de sustancias estimulantes como el café, las bebidas energizantes y las nuevas sustancias psicoactivas en una muestra de adolescentes italianos. En 2017 se realizó una encuesta en una muestra representativa de jóvenes italianos de 14 a 17 años, constituida por 15 833 estudiantes provenientes de 201 escuelas de educación secundaria. Se realizó un análisis de regresión logística para evaluar la asociación entre juego de riesgo/juego problemático (R1) y factores predictivos independientes: el modelo incluyó variables independientes (café, bebida energética y consumo de nuevas sustancias psicoactivas) y otras covariables demográficas, del entorno social y de conductas de riesgo. También se realizó un análisis de sensibilidad para examinar una segunda variable dependiente con respecto a cualquier experiencia de conductas relacionadas con el juego (R2). Los adolescentes clasificados como jugadores de riesgo o jugadores con problemas tenían una probabilidad significativamente mayor de consumir bebidas energizantes que los no jugadores o los jugadores sin riesgo. Se observó un patrón similar en el consumo de nuevas sustancias psicoactivas. No se evidenció ninguna asociación significativa con el consumo de café. El análisis de sensibilidad mostró que, en comparación con los no jugadores, el grupo de jugadores tenía mayores probabilidades de consumo frecuente de café, bebidas energéticas y/o nuevas sustancias psicoactivas. La evaluación del juego y el uso de sustancias estimulantes puede proporcionar información importante. Por consiguiente, podría ser necesario tomar medidas para reducir el uso de sustancias estimulantes como parte de los esfuerzos dirigidos a lidiar con los hábitos de juego poco saludables.

Pathways connecting socioeconomic variables, substance abuse and gambling behaviour: a cross-sectional study on a sample of Italian high-school students.

OBJECTIVES: The aim of this study was to elucidate the pathway of associations linking gambling, alcohol intake, smoking habit, cannabis consumption between each other and with demographic and socioeconomic variables. SETTING AND PARTICIPANTS: A survey was conducted in 2017 on a representative sample of 15 602 Italian 14-year-olds to 17-year-olds attending 201 secondary schools. OUTCOME MEASURES: Structural Equation Models analysis was used to assess the pathway between gambling, alcohol intake, smoking, cannabis consumption, demographics and socioeconomic factors. RESULTS: Irrespective of socioeconomic or demographic variables, gambling is positively associated with alcohol and cannabis consumption, while cannabis consumption is predicted by smoking and by alcohol intake, smoking is predicted by alcohol intake. Adolescents with a higher weekly income are more at risk of gambling, drinking alcohol and smoking, while the degree of economic dissatisfaction was positively associated with alcohol intake, cannabis consumption and smoking. Maternal employment appeared to be positively associated with adolescents' smoking, alcohol intake and cannabis consumption. CONCLUSIONS: This is one of the first studies to shed light on the pathways of associations connecting various health-risk behaviours among adolescents with demographic and socioeconomic factors.

Improvement of hair testing for Ethylglucuronide by supported liquid extraction and ultra-high performance liquid chromatography tandem mass spectrometry.

To improve the reproducibility, suitability and speed of hair testing for Ethylglucuronide (EtG), an ultra-high performance liquid chromatography coupled to tandem mass spectrometry (UHPLC-MS/MS) method was developed and validated together with a supported liquid extraction (SLE) EtG from the keratin matrix. EtG was analyzed using reversed phase chromatography with gradient elution and detection with tandem mass spectrometry operated in multiple reaction monitoring (MRM) mode via negative electrospray ionization (ESI). The method showed good linearity from limit of quantification (LOQ) to 100 pg/mg hair (r2 0.996 ±â€¯0.004). Recovery of the analyte was always higher than 80%, whereas intra- and inter-assay precision were always better than 15%. The developed method was applied to the analysis of more than 200 samples with medico-legal and epidemiological purposes ranging from non-detection of the analyte to 88.1 pg/mg and its robustness was proved by reanalysis of six different proficiency testing samples from the Society of hair testing obtaining a Z-score always less than 2.

Ultra-High-Performance Liquid Chromatography Tandem Mass Spectrometry Assay for Determination of Endogenous GHB and GHB-Glucuronide in Nails.

Background: The short chain fatty acid gamma-hydroxybutyric acid (GHB) is a precursor, and the metabolite of gamma-aminobutyric acid is commonly used as an illegal recreational drug of abuse. Methods: An ultra-high-performance liquid chromatography tandem mass spectrometry was developed and validated for endogenous GHB and its glucuronide in nails, to complement hair in forensic contexts for a retrospective detection of psychotropic drugs consumption. Results: GHB endogenous values for children and adolescents, adult females, and adult males in fingernails ranged from 0.3 to 3.0, 3.2, and 3.8 ng/mg, respectively, and toenails values ranged from 0.3 to 1.8, 2.0, and 2.4 ng/mg, respectively. In the three different groups, values of GHB in fingernails were statistically higher than those in toenails. GHB glucuronide could only be detected in finger nails with values ranging from 0.08 to 0.233, 0.252 and 0.243 in children and adolescents, adult females and adult males, respectively. Conclusions: The validated method was efficaciously applied to real finger and toe nails specimens from a population of males and females non GHB consumers. A preliminary cut-off of 5.0 ng/mg nail for endogenous GHB and 0.5 ng/mg for endogenous GHB-Gluc in the general population was proposed.

Determination of different recreational drugs in sweat by headspace solid-phase microextraction gas chromatography mass spectrometry (HS-SPME GC/MS): Application to drugged drivers.

A procedure based on headspace solid-phase microextraction (HS-SPME) coupled with gas chromatography/mass spectrometry (GC/MS) has been developed for the determination of most commonly used drugs of abuse in sweat of drivers stopped during roadside controls. DrugWipe 5A sweat screening device was used to collect sweat by a specific pad rubbed gently over forehead skin surface. The procedure involved an acid hydrolysis, a HS-SPME extraction for drugs of abuse but Δ(9)-tetrahydrocannabinol, which was directly extracted in alkaline medium HS-SPME conditions, a GC separation of analytes by a capillary column and MS detection by electron impact ionisation. The method was linear from the limit of quantification (LOQ) to 50ng drug per pad (r(2)≥0.99), with an intra- and inter-assay precision and accuracy always less than 15% and an analytical recovery between 95.1% and 102.8%, depending on the considered analyte. Using the validated method, sweat from 60 apparently intoxicated drivers were found positive to one or more drugs of abuse, showing sweat patches testing as a viable economic and simple alternative to conventional (blood and/or urine) and non conventional (oral fluid) testing of drugs of abuse in drugged drivers.

Gender disparity in addiction: an Italian epidemiological sketch.

INTRODUCTION: Gender disparity in different fields of addiction such as tobacco smoking, alcohol use, drugs of abuse consumption and doping practice has been investigated in Italian population. METHODS: We used the surveys and studies carried out for the above reported issues in recent years as revised by the "National Observatory on Tobacco smoke, Drugs of abuse, Alcohol and Doping" at Istituto Superiore di Sanità. RESULTS: Concerning tobacco habit, the trend of smoking women has been in constant decrease from a 19.7% in 2010 to a 16.9% in 2015, differently from men who passed from a 23.9% in 2010 to a 25.1% in 2015 with a slight increase in the habit. With respect to alcohol, in the last five years an increasing trend of consumption has been observed in 18-24 years old women, with 53% drinking women in the age range of 18-19 years overcoming the 50.4% general female population. Generally speaking, a one to four ratio can be underlined in the percentage of elderly women with a risky alcohol consumption with respect to men, while in case of adolescents and young adults gender disparity is not so pronounced. Drug abuse still remains a prevalent male phenomenon. However, an increase in cannabis users for both genders has been reported with a prevalence of "once in the life" around 20%, although more pronounced in females (+2.66 percentage points for females vs +0.93 percentage points in male). With respect to cocaine, the second most consumed drug, a reduction in consumption has been recently observed mainly in female population (-42.1%) than in men one (-27.5%). Finally, there are significant gender differences in doping attitude and/or in doping profiling. First of all, males seem to be more exposed to doping than females The prohibited substances most frequently used by females athletes are "Diuretics and Masking Agents" (38.3% positive female vs 14% males) compared to males athletes who use mostly anabolic agents (20.1% males vs 11.2% females). CONCLUSIONS: Results presented for the different fields of addiction show that a gender disparity is apparent and that females are less prone in having an addiction behaviour, although the young generation seems to increase that tendency.

Emerging trends in consuming behaviours for non-controlled substances by Italian urban youth: a cross sectional study.

INTRODUCTION: We investigated emerging trends in consuming behaviours for non-controlled substances in a cross sectional study on urban Italian adolescents and young adults, the reasons for consumption and risk perception as function of age, the relation with lifestyles and finally risk factors associated. METHODS: The survey methodology involved the administration of an anonymous questionnaire. It consisted of 68 questions, divided into five sections: personal details, socioeconomic characteristics, family and peer group, free time and lifestyles, and substances use. RESULTS: A total of 2621 adolescents and young adults (14-35 years old) from seven different Italian cities answered the questionnaire. The substances examined were mainly used recreationally (alcohol, energy drinks and smart-drugs) or to improve physical and sexual performance (physical performance-enhancing drugs, anabolic steroids and male sexual enhancement). The knowledge of the health related harm arising from the use of these products was very high for alcohol (> 90%), high for smart-drugs (> 70%), but significantly lower for anabolic steroids, drugs for sexual enhancement (~ 60%), physical performance-enhancing drugs or energy drinks (~ 55%). The principal risks factors for consumption were: the influence of friends (OR: 8.8), attending recreational places (OR: 5.4) aged between 25-35 years (OR: 3.0), be male (OR: 2.5) and having a bad relationship with the mother (OR: 2.1). CONCLUSIONS: These results have implication for prevention and early intervention programs of "physically and psychologically enhancing" non controlled substances use, which similarly to what is frequently advised for classical illicit drugs should focus on information campaigns and awareness initiatives especially addressed to young male adults who go clubbing, live outside the family and showed closed links with peers.

Segmental hair analysis to assess effectiveness of single-session motivational intervention to stop ethanol use during pregnancy.

BACKGROUND: This study aimed to test the effectiveness of single-session motivational intervention to stop ethanol use during pregnancy using segmental hair analysis of ethyl glucuronide to objectively verify drinking behavior before and after intervention. METHODS: 168 pregnant women attending Hospital del Mar (Barcelona, Spain) for antenatal visit were included in the study and randomly assigned to one of two conditions: single-session motivational intervention (MI; N=83) or single-session educational control condition (ECC; N=85). Ethyl glucuronide was measured in maternal hair divided into three segments of 3 cm each corresponding to the three different gestation trimesters by a validated liquid chromatography tandem mass spectrometry method. Concentrations of EtG<7 pg/mg, between 7 and 30 pg/mg and ≥30 pg/mg in each segment were used to assess total abstinence, repetitive moderate drinking and chronic excessive consumption in the previous three months. RESULTS: About a third of pregnant women self-reporting no ethanol consumption during gestation showed hair EtG values corresponding to ethanol drinking. Single-session MI helped in decreasing alcohol consumption during pregnancy as assessed by lower hair EtG concentrations in 2nd and 3rd trimesters. However, it did not significantly increase complete abstinence in pregnant women who previously showed hair EtG compatible with ethanol consumption. CONCLUSIONS: Pregnant women did not correctly self reported ethanol consumption during gestation, while hair EtG was essential to correctly identify drinking patterns. Single-session MI was not enough to stop ethanol use during pregnancy. Interventions at any visit during pregnancy are strongly recommended.

Pediatric exposure to drugs of abuse by hair testing: monitoring 15 years of evolution in Spain.

Hair testing is a useful tool to investigate the prevalence of unsuspected chronic exposure to drugs of abuse in pediatric populations and it has been applied to three different cohorts of children from Barcelona, Spain along fifteen years to evaluate eventual changes in this exposure. Children were recruited from three independent studies performed at Hospital del Mar (Barcelona, Spain) and approved by the local Ethics Committee. Hair samples were collected from the first 187 children cohort (around 4 years of age) in 1998, from the second 90 children cohort (1.5-5 years of age) in 2008 and from the third 114 children cohort (5-14 years of age) in 2013. Hair samples were analysed for the presence of opiates, cocaine, amphetamines, and cannabis by validated methodologies using gas or liquid chromatography-mass spectrometry. Familiar sociodemographics and eventual consumption of drugs of abuse by parents, and caregivers were recorded. Hair samples from 24.6% children in 1998 were positive for any drug of abuse (23.0% cocaine), 25.5% in 2008 (23.3% cocaine), and 28.1% in 2013 (20.1% cocaine and 11.4% cannabis). In none of the cohorts, parental sociodemographics were associated with children exposure to drugs of abuse. The results of the three study cohorts demonstrated a significant prevalence of unsuspected pediatric exposure to drugs of abuse which mainly involved cocaine maintained along fifteen years in Barcelona, Spain. We recommend to be aware about unsuspected passive exposure to drugs of abuse in general population and to use general or selected hair screening to disclose exposure to drugs of abuse in children from risky environments to provide the basis for specific social and health interventions.

The NASOROSSO (Rednose) project: an Italian study on alcohol consumption in recreational places.

The Nasorosso project of the Italian Youth Department and the National Institute of Health, aimed to raise awareness about drinking and driving under the influence of alcohol among club goers with a series of initiatives. Within the framework of the project, blood alcohol concentration (BAC) was measured on 106,406 individuals before and after clubbing in 66 different recreational sites from 11 Italian provinces, over 16 months. Participating individuals were interviewed regarding sociodemographic and environmental characteristics and alcohol intoxicated people were offered to be taken home. The BAC median at the club entry was 0.26 g/L with 65.3% subjects showing a BAC value under the driving legal limit of 0.5 g/L. At the exit from clubs, BAC median value rose to 0.44 g/L and subjects with BAC value under the legal limit decreased to 54.9%. Being male, aged between 18 and 34 years with a diploma, being a drinker and entering the disco with a BAC already beyond the legal limit predicted a BAC value beyond 0.5 g/L at exit from the recreational place. Conversely, being a driver, being a student and exiting from the disco before 4 a.m. reduced the probability of having a BAC higher than 0.5 g/L at the end of the night. Health policies to prevent harmful use of alcohol in young people should continue to offer targeted information/ prevention; in order to steadily increase the awareness of the dangers and the damages of excessive use of alcohol.

Hair and urine testing to assess drugs of abuse consumption in couples undergoing assisted reproductive technology (ART).

For the first time in Europe hair and urine testing have been applied to assess drugs of abuse consumption in couples undergoing assisted reproductive technology and the eventual association of toxic habits with other lifestyle, health status and sociodemographic factors was also investigated. Couples attending five assisted reproduction centers in Rome were invited to join the study. When they presented at the Centre for the visit, they were asked to answer a structured questionnaire concerning sociodemographic characteristics and lifestyle habits, and at the same time to provide hair and urine samples. Hair and urine testing for drugs of abuse, urinary profile of principal endogenous steroids involved in fertility process (testosterone, epitestosterone, androsterone, etiocholanolone and dehydroepiandrosterone) and of alcohol and tobacco smoke biomarkers were performed with validated methodologies. Of the 594 enrolled individuals (297 couples), 352 (164 couples and 24 single individuals from the couple) completed the questionnaire and gave both hair and urine samples, apart from 3 bald men, who only gave urine samples. Urine testing showed an overall 4.8% (17 individuals) positivity to drugs of abuse: 4.2% to cannabinoids, 1.4% to cocaine and 0.85% to both drugs. Results of 4cm segment hair samples testing matched those from urine samples. Thus, taking together, results of urine and hair testing confirmed repeated use of cannabis, cocaine and both drugs in 3.7, 0.85 and 0.57% examined individuals, respectively. Drug consumers were in a statistically higher percentage active smokers and alcohol drinkers, less prone to physical activity and with a trend towards higher weight than non consumers. Finally, repeated drug consumption was associated with significant lower concentration of urinary testosterone in males and of urinary dehydroepiandrosterone in females. The findings of the present study confirm the suitability of urine testing to disclose recent drugs of abuse consumption and of hair analysis to verify repeated consumption. Association between different toxic habits and sedentary lifestyle is also substantiated by the obtained results in our cohort of couples attending assisted reproduction centers.

Development and validation of a liquid chromatography-tandem mass spectrometry assay for hair analysis of atomoxetine and its metabolites: Application in clinical practice.

Atomoxetine (ATX) is a potent inhibitor of the noradrenaline reuptake transporter approved since 2002 for the treatment of attention-deficit/hyperactivity disorder (ADHD) in children, adolescents, and adults as alternative treatment to methylphenidate. A procedure based on liquid chromatography-tandem mass spectrometry (LC-MS/MS) has been developed for the determination of ATX and its main metabolites (4-hydroxyatomoxetine - 4 hydroxyATX - and N-desmethylatomoxetine - des-methylATX) in hair of one treated child and five treated adolescents. Since hair samples can be easily collected without the need for specials skills and exposing a patient to discomfort, hair testing of ATX and eventually of its metabolites should be useful, especially in case of pediatric patients, to check compliance in a wider time-window. After addition of duloxetine as internal standard, hair samples were overnight digested with 2ml 1M NaOH at 45°C. Then, analytes were extracted from alkaline solution with two different 2ml aliquots of tert-butyl methyl ether. Chromatographic separation was achieved at ambient temperature using a reverse-phase column and a mobile phase of 40% of water-60% 5mM ammonium acetate, 50mM formic acid, 4mM trifluoroacetic acid in acetonitrile-water (85:15, v/v). The mass spectrometer was operated in positive ion mode using multiple reaction monitoring. The method was linear over the concentration range 0.2-50ng/mg hair for the all analytes under investigation, with an intra- and inter-assay imprecision and inaccuracy always less than 20% and an analytical recovery between 33.1% and 76.1%, depending on the considered analyte. Only ATX and 4-hydroxyATX were detected in hair samples with concentrations varying from 0.2 to 2.0ng/mg hair and from 0.3 to 1.0ng/mg, respectively. Notwithstanding the absence of any dose-hair concentration relationship, hair monitoring of ATX and concomitant medications commonly administrated in ADHD children and adolescents can be crucial in verifying long-term compliance to prescribed medication in individuals displaying a non negligible tendency to refuse drugs and to lie on the adherence to therapy as a specific symptom of the disease.