Testing ethylglucuronide in maternal hair and nails for the assessment of fetal exposure to alcohol: comparison with meconium testing.

BACKGROUND: The deleterious effects exerted by prenatal ethanol exposure include physical, mental, behavioral, and/or learning disabilities that are included in the term fetal alcohol spectrum disorder. The measurement of ethylglucuronide (EtG) in alternative biological matrices, including neonatal and maternal hair, neonatal meconium, and maternal nails, is receiving increasing interest for the accurate evaluation of the in utero exposure to alcohol. OBJECTIVE: To evaluate the correlation between EtG in maternal hair and nails with EtG in neonatal meconium to further explore the suitability of these biomarkers in disclosing prenatal exposure to ethanol. METHODS: A total of 151 maternal hair strands (0-6 cm), nail clips (2-6 mm), and corresponding neonatal meconium and nails samples were obtained from neonatal wards of 4 Mediterranean public hospitals: Rome, Florence, and Belluno in Italy and Barcelona in Spain. Hair, nails, and meconium were analyzed for the presence of EtG by validated liquid chromatography mass spectrometry assay. Meconium was also analyzed for the presence of fatty acid ethyl esters (FAEEs) as a complementary biomarker of potential in utero exposure to alcohol. RESULTS: Eighteen newborns resulted in utero exposed to maternal alcohol consumption by FAEE testing in meconium with EtG values between 0.5 and 1.5 nmol/g. Unfortunately, none of these cases were confirmed by the presence of EtG in maternal hair and nails samples, which resulted all negative to this biomarker. DISCUSSION AND CONCLUSIONS: The results confirm that FAEEs and EtG in meconium are the best biomarkers to assess in utero exposure to maternal alcohol. EtG in hair and nails are not good biomarkers to disclose alcohol consumption lower than on daily basis and lower than 1-2 alcoholic units per day.

Ethyl-glucuronide and ethyl-sulfate in placental and fetal tissues by liquid chromatography coupled with tandem mass spectrometry.

The aim of this study was to develop a method for the determination of ethyl-glucuronide (EtG) and ethyl-sulfate (EtS), two direct ethanol metabolites, in early placental and fetal human tissues, as potential biomarkers of transplacental ethanol transfer from the mother to the fetus. Placental and fetal tissue samples were obtained from women undergoing voluntary termination of pregnancy at 12 weeks of gestation. Samples were deproteinized and directly injected into a liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) system. Limits of detection of 13.0 and 23.0 pmol/g and lower limits of quantification of 22.0 and 40.0 pmol/g were reached for EtG and EtS, respectively. Inter- and intraday imprecision and accuracy were always lower than 15%. The method was applied to 70 samples (35 placentas and 35 fetal tissues). Of 35 samples, 4 samples collected from 4 women tested positive for EtG and EtS, always showing higher concentrations for EtG. The placenta/fetal tissue ratio for EtG was 2.9 ± 0.9, whereas EtS showed a ratio of 1.7 ± 0.7. Preliminary results suggest that these metabolites are present in both tissues. Further studies should now corroborate the hypothesis, not yet confirmed, that transplacental transfer of ethanol takes place not only for the parent compound but also for EtG and EtS.

Chronic excessive alcohol consumption diagnosis: comparison between traditional biomarkers and ethyl glucuronide in hair, a study on a real population.

BACKGROUND: The study aimed to evaluate the diagnostic sensitivity (SE) and specificity (SP) of chronic alcohol misuse diagnosis by comparing traditional biomarkers with ethyl glucuronide (EtG), an ethanol direct metabolite, detected in the keratinic matrix. METHODS: Seventy-six subjects tested for chronic alcohol abuse for different purposes were recruited. EtG was detected in hair, whereas the analyses of carbohydrate-deficient transferrin (CDT), alanine transaminase, aspartate transaminase, gamma-glutamyl transferase, mean corpuscular volume, and mean corpuscular hemoglobin were performed in serum samples. RESULTS: Of the 76 patients examined, 26 were judged by the medical doctors as subjects with alcohol abuse problems and, therefore, not eligible for driving license renewal or liver transplant. EtG in hair (SE = 0.68, SP = 1.00) showed the best diagnostic SE and SP compared with the other biomarkers investigated. Among the traditional biomarkers, only CDT proved to be suitable for forensic purposes because of the high diagnostic specificity (SP = 1.00) although it showed poor diagnostic SE (0.27). The percentage of positive samples decreased for all the biomarkers by excluding the subjects with hepatic diseases, except for EtG and CDT, suggesting that these 2 biomarkers could be less affected by false positive results, because of hepatic diseases. CONCLUSIONS: This study showed that when EtG in hair and CDT results are combined, diagnostic SE in chronic alcohol abuse diagnosis clearly improved, suggesting that complementary analysis of both these biomarkers provides the best diagnostic tool in suspected cases of chronic excessive alcohol consumption.

Comparison between two orthodontic skeletal anchorage devices: osseointegrated implants and miniscrews - Medical-Legal Considerations.

The object of this article is to describe potential medical-legal problems concerning the use of miniscrews as orthodontic skeletal anchorage. The miniscrews, which are already used in rigid fixation, do not need either a healing period before loading, or complete osseo-integration as do implants. Their particular shape allows high primary stability and they can be placed in the sub-periosteal region. The dimensions of these miniscrews are minute, with dimensions of only 4-5 mm in length. They penetrate only slightly deeper than the cortical layer and therefore notably reduce the risk of lesions to roots, nerves, or maxillary sinus, when compared to other skeletal anchorage systems such as implants. The use of the miniscrew in oral surgery or periodontology does not need specific insurance coverage in contrast to the insertion of implants. Accordingly, orthodontic therapy completed by the dentist with orthopaedic microscrews as anchorage could also be included in a general civil insurance policy for orthodontics.

Observations on dental structures when placed in contact with acids: experimental studies to aid identification processes.

In a "mafia" crime case, a magistrate asked us whether it is possible to destroy a cadaver by immersing it in acids, and would it be possible to identify any residues. The aim of this study was to observe the behavior of teeth exposed to four kinds of acid solutions. The teeth were placed in plastic containers with 25 mL of acid and observed. The experiences showed that teeth are completely dissolved after 14 h of immersion in 37% solution of hydrochloric acid, while at 90h in 96% sulfuric acid, the destruction of the samples is still incomplete. In nitric acid the teeth undergo a complete dissolution in 12 h, and in 17 h in aqua regia (chloroazotic acid-hydrochloric/nitric acid 1:3). It was possible to recognize the characteristic morphological features of dental tissues and structures up until the advanced stages of degradation.

Determination of perfluorooctanoic acid and perfluorooctanesulfonate in human tissues by liquid chromatography/single quadrupole mass spectrometry.

A method is described that permits the measurement of the levels of perfluorooctanoic acid (PFOA) and perfluorooctane sulfonate (PFOS) in human liver, kidney, adipose tissue, brain, basal ganglia, hypophysis, thyroid, gonads, pancreas, lung, skeletal muscle and blood, even in subjects not occupationally exposed to these compounds. The purification of samples involved the use of trifunctional (tC18) and strong anion-exchange (SAX) solid-phase extraction cartridges, and the analysis utilized a high-performance liquid chromatograph coupled to a single quadrupole mass spectrometer (LC/MS). The analyses were conducted on a mixed-bed reversed-phase column by gradient runs using 3 mM ammonium acetate/methanol mixtures at different proportions as the mobile phase. The detector was used in electrospray negative ion mode by recording simultaneously the ions m/z 413.0 (PFOA) and 499.0 (PFOS). Perfluorononanoic acid (PFNA), added to the samples before the purification, was used as the internal standard (ion monitored = m/z 463.6). The recovery rates of the extraction procedure ranged from 79.6 to 95.6% (CV% 1.7-7.4%) for PFOA, from 79.7 to 100.8% (CV% = 1.2-7.1) for PFOS, and from 89.1 to 102.3% (CV% = 0.9-5.2 %) for PFNA. The calibration curves were linear up to at least 400 ng of analytes per gram of tissue. The detection limits (signal-to-noise ratio = 3) were 0.1 ng/g for both PFOA and PFOS measured in all tissues except adipose tissue, where the limits were about 0.2 ng/g. The content of analytes in tissues varied from 0.3 to 3.8 ng/g (respectively: basal ganglia and lung) for PFOA, and from 1.0 to 13.6 ng/g (respectively: skeletal muscle and liver) for the linear isomer of PFOS. The method is suitable to evaluate the content of PFOA and PFOS in different tissues taken from the general population exposed to very low concentrations of these pollutants.