Rosanna Mancinelli

Simultaneous liquid chromatographic assessment of thiamine, thiamine monophosphate and thiamine diphosphate in human erythrocytes : a study on alcoholics

An isocratic HPLC procedure for the assessment of thiamine (T), thiamine monophosphate (TMP) and thiamine diphosphate (TDP) in human erythrocytes is described. Several aspects of the procedure make it suitable for both clinical and research purposes: limits of detection and quantification of 1 and 2.5 nmol/l, respectively, recovery of 102% on average (range 93-112%), intra- and inter-day precisions within 5 and 9%, respectively, total elution time 15 min. This analytical methodology was applied to a case-control study on erythrocyte samples from 103 healthy subjects and 36 alcohol-dependent patients at risk of thiamine deficiency. Mean control values obtained were: T=89.6+/-22.7 nmol/l, TMP=4.4+/-6.6 nmol/l and TDP=222.23+/-56.3 nmol/l. T and TDP mean values of alcoholics were significantly lower than those of control cases: T=69.4+/-35.9 nmol/l (P<0.001) and TDP=127.4+/-62.5 nmol/l (P<10(-5)). The diagnostic role of TDP was evaluated and a significant role for thiamine was established in the study of alcohol related problems.

Early exposure to ethanol but not red wine at the same alcohol concentration induces behavioral and brain neurotrophin alterations in young and adult mice

Ethanol exposure during pregnancy is one of the major causes of mental retardation in western countries by inducing fetal-alcohol-like-syndromes. Red wine is known to contain ethanol but also compounds with putative antioxidant properties. It has also been shown that nerve growth factor (NGF) and brain derived neurotrophic factor (BDNF) are severely affected by ethanol during prenatal and postnatal life. The aim of the current study was to investigate in male CD1 mice brain alterations in NGF and BDNF due to chronic early exposure to ethanol solution (11 vol%) or to red wine at the same alcohol concentration starting from 60 days before pregnancy up to pups weaning. Data revealed no differences between groups of dams in pregnancy duration, neither in pups delivery, pups mortality and sex ratio. Data also showed that adult animals exposed to only ethanol had disrupted levels of both NGF and BDNF in the hippocampus and other brain areas. This profile was associated with impaired ChAT immunopositivity in the septum and Nuclei Basalis and with altered cognition and emotional behavior. Quite interestingly mice exposed to red wine had no change in the behavior or in ChAT immunopositivity but a decrease in hippocampal BDNF and a mild NGF decrease in the cortex. Also NGF-induced neuritic outgrowth in PC-12 cells was still present when exposed to red wine but not when exposed to ethanol solution only. Data suggest differences in ethanol-induced neurotoxicity between red wine and ethanol solution only.

Biomarkers in Alcohol Misuse: Their Role in the Prevention and Detection of Thiamine Deficiency

In Western countries alcohol misuse is the most frequent cause of thiamine (vitamin B1) deficiency (TD) and consequent neuro-impairment. Studies have demonstrated that between 30 and 80% of alcoholics are thiamine deficient, and this puts them at risk of developing the Wernicke-Korsakoff (WK) syndrome. The relative roles of alcohol and TD in causing brain damage remain controversial and it is important to try to determine the role played by each factor. Animal studies support an additive effect of alcohol exposure and TD, and indicate the potential for interaction between alcohol and TD in human alcohol-related brain damage. Early diagnosis of alcohol-related TD is therefore an important aspect of effective intervention and treatment. Alcohol biomarkers provide a direct and indirect way of estimating the amount of alcohol being consumed, the duration of ingestion and the harmful effects that long-term alcohol use has on body functions. Appropriate use of these markers is very helpful when considering a diagnosis of alcohol-related TD.

The Chemical Components of Electronic Cigarette Cartridges and Refill Fluids: Review of Analytical Methods

INTRODUCTION To date, several concerns have been raised on the purity of ingredients employed in the manufacturing processes of refill fluids and cartridges, the device functionality, and the quality control of electronic cigarettes. This article reviews analytical methods so far described for the analysis of liquids to detect their chemical components and to investigate the presence of toxicants and carcinogens that can potentially occur as impurities of ingredients or as a consequence of their degradation. RESULTS AND DISCUSSION Based on the scientific literature, high-performance liquid chromatography with diode-array detection (HPLC/DAD) and liquid chromatography-tandem mass spectrometry (LC-MS/MS) are most appropriate for determining nicotine and related compounds in fluids and cartridges, whereas LC-MS/MS has been successfully used to determine nitrosamines. Content analyses of glycols have been performed using gas chromatography equipped with flame ionization detector or gas chromatography/mass spectrometry (GC/MS), whereas carbonyl and other volatile organic compounds determinations have been performed by HPLC/DAD and GC/MS, respectively. Content analyses of heavy metals have been performed by inductively coupled plasma optical emission spectroscopy or inductively coupled plasma mass spectrometry. Since new potentially toxic substances may be created during heating, it is also necessary to investigate the chemical composition of generated aerosol. In this case, similar methods applied for tobacco smoke can be adopted. CONCLUSIONS A broad range of analytical techniques are available for the detection of constituents and toxicants in e-liquids and cartridges. Analyses of liquids have been performed with pharmacopeia procedures and methods (International Organization for Standardization, Environmental Protection Agency, and American Public Health Association) developed for other matrices but applicable to e-liquids. Because new potentially harmful substances may be produced during heating process, analyses of aerosol are needed to correlate its composition to the chemical components of liquids.

Early exposure to ethanol or red wine and long-lasting effects in aged mice. A study on nerve growth factor, brain-derived neurotrophic factor, hepatocyte growth factor, and vascular endothelial growth factor

Prenatal ethanol exposure produces severe changes in brain, liver, and kidney through mechanisms involving growth factors. These molecules regulate survival, differentiation, maintenance, and connectivity of brain, liver, and kidney cells. Despite the abundant available data on the short and mid-lasting effects of ethanol intoxication, only few data show the long-lasting damage induced by early ethanol administration. The aim of this study was to investigate changes in nerve growth factor (NGF), brain derived neurotrophic factor (BDNF), hepatocyte growth factor (HGF), and vascular endothelial growth factor (VEGF) in brain areas, liver, and kidney of 18-mo-old male mice exposed perinatally to ethanol at 11% vol or to red wine at the same ethanol concentration. The authors found that ethanol per se elevated NGF, BDNF, HGF, and VEGF measured by ELISA in brain limbic system areas. In the liver, early exposure to ethanol solution and red wine depleted BDNF and VEGF concentrations. In the kidney, red wine exposure only decreased VEGF. In conclusion, the present study shows that, in aged mice, early administration of ethanol solution induced long-lasting damage at growth factor levels in frontal cortex, hippocampus, and liver but not in kidney. Otherwise, in mice exposed to red wine, significant changes were observed in the liver and kidney but not in the hippocampus and frontal cortex. The brain differences in ethanol-induced toxicity when ethanol is administered alone or in red wine may be related to compounds with antioxidant properties present in the red wine.

Clinical delineation of fetal alcohol spectrum disorders (FASD) in Italian children: comparison and contrast with other racial/ethnic groups and implications for diagnosis and prevention.

In Italy, little is known about the spectrum of adverse fetal effects related to maternal alcohol use during pregnancy. In this paper, we report on the phenotype of Italian children with fetal alcohol spectrum disorders (FASD). These data were gathered as part of a field study assessing the prevalence of FASD in children in an in-school study in a rural area near Rome. The purposes of this paper are: (1) to completely characterize the clinical phenotype of a large cohort of Italian children with FASD; (2) to correlate and contrast the phenotype of this population with that observed in other populations and reported in the medical literature; (3) to discuss the drinking habits of Italian women, before, during and after pregnancy; and (4) to suggest mechanisms for intervention and prevention of FASD based on data gathered from this study.

Simple and reliable high-performance liquid chromatography fluorimetric procedure for the determination of amphetamine-derived designer drugs.

The paper describes a HPLC-fluorimetric procedure for the determination of methylenedioxyamphetamine, methylenedioxymethamphetamine, methylenedioxyethamphetamine and N-methyl-1-(1,3-benzodioxol-5-yl)-2-butanamine in urine, serum, saliva and street samples, that features interesting advantages over other procedures previously described. The method requires a very small sample volume (100 microl) and no extraction, lacks matrix effect, and is not time consuming. Linearity was in the range 50-1000 ng/ml regardless of matrix. Sensitivity and detection limit were 50 ng/ml and 10 ng/ml, respectively, but they may reach 10 ng/ml and 2 ng/ml if a slight modification is introduced in the procedure. Intra- and inter-day precision were always within 5% and 8%, respectively. Recovery was satisfactory for all matrices. The described procedure could be successfully used for clinical, epidemiological and forensic applications.

Woman, alcohol and environment: Emerging risks for health

Alcohol drinking is one of the most relevant problems in Western Countries but the negative effects of alcohol misuse are often neglected or underestimated with serious consequences for public health. Over the last few years a rapid growth in the number of drinking females and the decrease of their age of first use, have increased the health risk for women and their offspring. Moreover, modern environments facilitate pollutants exposure, further escalating the health risks due to lifestyle habits. This review takes into account the peculiarities of alcohol effects on female health and the risks of teratogenic effects. The possible interaction between alcohol and pollutants exposure is also discussed. The role of biomarkers against alcohol-related damage is presented as an invaluable clinical tool, including early intervention, treatment monitoring and, above all, prevention of prenatal non-reversible damage. Recent alcohol studies show the greater severity of alcohol damage in female subjects and the need of gender-targeted intervention.

Blood thiamine, zinc, selenium, lead and oxidative stress in a population of male and female alcoholics: clinical evidence and gender differences

INTRODUCTION Long term alcohol abuse is associated with deficiencies in essential nutrients and minerals that can cause a variety of medical consequences including accumulation of toxic metals. AIM The aim of this research is to get evidence-based data to evaluate alcohol damage and to optimize treatment. Thiamine and thiamine diphosphate (T/TDP), zinc (Zn), selenium (Se), lead (Pb) and oxidative stress in terms of reactive oxygen metabolites (ROMs) were examined in blood samples from 58 alcohol dependent patients (17 females and 41 males). RESULTS T/TDP concentration in alcoholics resulted significantly lower than controls (p < 0.005) for both sexes. Serum Zn and Se did not significantly differ from reference values. Levels of blood Pb in alcoholics resulted significantly higher (p < 0.0001) than Italian reference values and were higher in females than in males. ROMs concentration was significantly higher than healthy population only in female abusers (p = 0.005). CONCLUSION Alcoholics show a significant increase in blood oxidative stress and Pb and decrease in thiamine. Impairment occurs mainly in female abusers confirming a gender specific vulnerability.

Paternal alcohol exposure in mice alters brain NGF and BDNF and increases ethanol-elicited preference in male offspring

Ethanol (EtOH) exposure during pregnancy induces cognitive and physiological deficits in the offspring. However, the role of paternal alcohol exposure (PAE) on offspring EtOH sensitivity and neurotrophins has not received much attention. The present study examined whether PAE may disrupt nerve growth factor (NGF) and/or brain‐derived neurotrophic factor (BDNF) and affect EtOH preference/rewarding properties in the male offspring. CD1 sire mice were chronically addicted for EtOH or administered with sucrose. Their male offsprings when adult were assessed for EtOH preference by a conditioned place preference paradigm. NGF and BDNF, their receptors (p75NTR, TrkA and TrkB), dopamine active transporter (DAT), dopamine receptors D1 and D2, pro‐NGF and pro‐BDNF were also evaluated in brain areas. PAE affected NGF levels in frontal cortex, striatum, olfactory lobes, hippocampus and hypothalamus. BDNF alterations in frontal cortex, striatum and olfactory lobes were found. PAE induced a higher susceptibility to the EtOH rewarding effects mostly evident at the lower concentration (0.5 g/kg) that was ineffective in non‐PAE offsprings. Moreover, higher ethanol concentrations (1.5 g/kg) produced an aversive response in PAE animals and a significant preference in non‐PAE offspring. PAE affected also TrkA in the hippocampus and p75NTR in the frontal cortex. DAT was affected in the olfactory lobes in PAE animals treated with 0.5 g/kg of ethanol while no differences were found on D1/D2 receptors and for pro‐NGF or pro‐BDNF. In conclusion, this study shows that: PAE affects NGF and BDNF expression in the mouse brain; PAE may induce ethanol intake preference in the male offspring.