Effects of various sample pretreatment procedures on ethyl glucuronide quantification in hair samples: Comparison of positivity rates and appraisal of cut-off values.

The quantification of ethylglucuronide (EtG) in hair is nowadays recognized as the approach with the highest diagnostic performance to evaluate harmful drinking. A widely accepted cut-off of 30pg/mg has been selected after several accurate compared studies. While most of the studies that were used to establish the appropriate cut-off value prescribed to cut hair into small segments before their extraction, hair milling has subsequently been identified as the most efficient pretreatment procedure and was therefore recommended in the last Consensus document issued by the Society of Hair Testing. In this study, we initially compared the results obtained with the two sample preparations, namely cutting and milling, both being applied to the same specimens (n=781). Among these, 205 samples produced measurable EtG values with both methods, with differences ranging from -41.7% up to +415% (the mean increase in EtG concentration, switching from cutting to milling, was +62.1% and the median was +42.3%). Among the aforementioned 205 samples, 29 specimens (3.7% of the total 781 samples) produced significantly different outcome, being classified as negative (i.e., below 30pg/mg) if the cutting procedure is used, but largely positive (above 40pg/mg) when milling is used. Subsequently, the positivity rates obtained on a large population dataset (>27,000 samples) with the two procedures, were retrospectively compared using variable cut-offs values. The percentage of head hair samples with EtG concentration exceeding 30pg/mg upon application of the milling procedure shows a 45% increase (from 10.9% to 15.8%) with respect to cutting procedure, whereas the fraction of hair samples with EtG exceeding 40pg/mg (10.5%) overlaps the percentage of positive samples obtained after cutting pretreatment and applying a cut-off of 30pg/mg. On the basis of these results, it would be worth considering the application of cut-off values linked with the pretreatment procedure, taking into account the results of forthcoming inter-laboratory calibrations.

Characterisation of CIME, an experimental chamber for simulating interactions between materials of the cultural heritage and the environment.

An approach consisting in combining in situ and laboratory experiments is often favoured for investigating the mechanisms involved in the weathering of the materials of the cultural heritage. However, the realistic simulation in the laboratory of the environmental conditions ruling the interactions of atmospheric compounds with materials is a very complex task. The aim of this work is to characterise CIME, a new chamber specially built to simulate the interactions between materials of the cultural heritage and the environment. The originality of this instrument is that beside the usual climatic parameters (temperature, relative humidity, solar radiation) and gaseous pollutants, it also allows the controlled injection of different types of particulate matter such as terrigenous, marine and anthropogenic. Therefore, varied realistic atmospheric environments (marine or urban) can be easily simulated within CIME. In addition to the technical description of CIME, this paper shows the first results obtained by the impact of gaseous pollutants on non-durable glass, bronze and limestone. The first experiments for the deposition of different particles (calcite, clays, soot and halite) are also presented.

Physico-chemical characterisation of glass soiling in rural, urban and industrial environments.

Glass materials are broadly used in the built environment (windows, facades, roofs, museum showcases, and solar panels) due to their optical (transparency) and thermal properties. Their interaction with the multiphase atmospheric medium results in a more or less pronounced transparency loss called soiling. This phenomenon leads to a loss of amenity of artefacts; consequently, high cleaning costs have to be supported by public and private entities. Complete understanding of the nature of surface deposit appears thus extremely important for addressing strategies to control it. The present research is based on the sheltered exposure, in different environments, of durable glass panels during 1 year. At these different locations, airborne pollutant concentrations have also been monitored. Three environments have been investigated: rural (R), urban (U) and industrial (I). Results show that the mass of the deposit and the optical impairment of the glass (haze) are too spread to allow discriminating between different environments. However, the analyses of soluble species and particulate organic matter allow identifying factors responsible for soiling and highlighted the reactivity of deposit to relative humidity which favours post-deposit evolution.

Isolation and culture of pig tonsil lymphocytes.

Tonsils are secondary lymphoid organs that play an important role in host defense. The aim of our study was to develop reliable procedures for isolation and culture of pig tonsil cells, and to validate their possible use in functional immunoassays. Using our isolation procedure, we recovered on average 238.7 ± 107.1 × 10(6) cells per tonsil couple with a mean vitality of 89.8 ± 2.7%. These values significantly decreased 8 months after freezing at -80°C along with the subsequent spontaneous release of both IgA and IgG in culture. These results suggest to use pig tonsil cells within 2 months from thawing to maintain suitable conditions in terms of recovery, vitality and release of antibody in vitro. Tonsil mononuclear cells also showed the ability to secrete antimicrobial peptides and to respond in vitro to immunological stimuli. On the whole, our study has defined operating conditions for tonsil processing, control of bacterial contaminations, time limits of storage at -80°C, as well as for evaluating polyclonal Ig production in vitro. Such procedures are likely to be of some importance in studies on regional immunity and in the development of large animal models for biomedical sciences.

Dose-response function for the soiling of silica-soda-lime glass due to dry deposition.

Several exposure campaigns of silica-soda-lime window glass have been performed in 30 European sites and 1 in Canada in order to understand, quantify and model the phenomenon of soiling. In this purpose samples were exposed sheltered from the rain. Parallel to exposure, several meteorological parameters and pollution concentrations have been monitored. This paper shows first results on the establishment of a dose-response function for glass soiling. Statistical analyses show that PM(10) is not the only parameter, but also SO(2) and NO(2) atmospheric concentrations seem to be responsible for the optical impairment of glass surfaces, expressed as haze.

Altered osteoprotegerin/RANKL ratio and low bone mineral density in celiac patients on long-term treatment with gluten-free diet.

Skeletal demineralization and mineral metabolism derangement are well-recognized features of untreated celiac disease (CD). Although treatment with a gluten-free diet appears to prevent bone loss while correcting skeletal demineralization in childhood, there is evidence that bone mineral density does not return to normal in celiacs diagnosed in adulthood. Osteoprotegerin (OPG), a member of the tumor necrosis factor receptor family, and ligand of receptor activator of NFkB (RANKL) are involved in the process of bone turnover and have been implicated in the pathogenesis of osteoporosis and other metabolic bone diseases. We measured OPG, RANKL, bone mineral density (BMD), and biochemical markers of bone turnover in 32 adult female premenopausal celiac patients on a gluten-free diet, and thirty age-matched healthy women. We correlated the OPG/RANKL ratio with the severity of bone loss. Celiac patients had a mean BMD lower than controls in lumbar spine and in the femoral neck. Serum levels of bone alkaline phosphatase (BAP, marker of bone formation), and urinary excretion of telopeptides of type I collagen (a marker of bone resorption) were significantly higher than in controls. Serum OPG and RANKL levels were significantly higher in CD patients than in controls, while the OPG/RANKL ratio was significantly lower in CD patients than in controls and was positively correlated with BMD at the spine. The role of elevated OPG in CD patients is unclear, but it might represent a compensatory mechanism against other factors that promote bone damage. Further studies are required to assess a possible therapeutic potential of osteoprotegerin in optimally treated celiacs with persistent osteopenia.

Modeling of soiling based on silica-soda-lime glass exposure at six European sites.

Samples of silica-soda-lime float glass, a material selected as a pertinent soiling sensor, were exposed for up to 28 months at four urban sites (Athens, Krakow, London and Prague) and at a semi-urban one (Monte Libretti, near Rome), sheltered from rain. This exhaustive experience permitted to complete and to test the generalisation ability of the results previously obtained on the same material, exposed according to the same protocol, during 24 months, at a single site (Paris). The model previously fitted for the Paris experiment, the Hill equation, could be successfully generalised for four other sites of exposure (Athens, Krakow, Prague and Rome). The analytical form of this model was interpreted in terms of a physical description of the soiling phenomenon. Some of the model coefficients were different from one site to another, depending on the specific environment of exposure (air pollution levels, meteorological factors), while the other ones were rather equivalent. The analysis of the model coefficients, on the one hand, led to an estimation of the period after which soiling is close to its saturation level, and on the other hand, it will permit to correlate these coefficients to the environmental factors, in order to select the most appropriate ones for building dose-response functions for soiling, with broad geographical application.

Thalassaemic men affected by erectile dysfunction treated with transurethral alprostadil: case report.

The aim of this controlled clinical study, performed in a specialized institutional unit for thalassaemic men, was to consider the possibility of restoring erection in beta-thalassaemic patients with erectile dysfunction by administering E(1) prostaglandins (alprostadil) transurethrally. Four patients affected by beta-thalassaemia, aged between 32 and 52 years, and having an erectile dysfunction were included in the study. Each patient was given 500 microg alprostadil in the distal urethra. Response was evaluated by the erection assessment scale. The main outcome measures were: (i) the clinical study; (ii) FSH, LH, total and free testosterone plasma concentrations; and (iii) basal and dynamic Doppler sonography of cavernous arteries. The treatment produced a response of 3-4 on the erection assessment scale. Average minimum response time was 20 min, while average maximum response time was about 60 min. There was no evidence of significant side effects. Our hypothesis is that the delayed reaction was due to organ damage induced by iron load, causing a reduction or absence of elasticity in the interstitial tissue of the corpora cavernosa. Thus, we believe that treatment with alprostadil can be considered an effective, non-invasive therapy for thalassaemic patients with erectile dysfunction.

Effects of smokeless tobacco-related sensory and behavioral cues on urge, affect, and stress.

The hypothesis that smokeless tobacco-related sensory and behavioral cues can act as conditioned stimuli was tested in a counterbalanced double-blind experimental design. The nicotine content of snuff smokeless tobacco (ST) was manipulated for 24 male ST users by mixing ST with an ST substitute. Affect was manipulated through imagery scripts, stress was induced by a mental arithmetic task, and physiological measures and self-reported affect, stress, and urge for ST were collected. Urge for ST was consistently reduced regardless of the nicotine content in the ST conditions. Urge was increased by the stress manipulation and by negative affect when compared with positive affect. Urge for ST was positively correlated with stress and negative affect but was not correlated with positive affect. Physiological arousal was not related to urge. Results generally parallel studies of smoking and suggest that ST substitute products may aid ST cessation.

Caffeine, but not time of day, increases whole-arm physiological tremor in non-smoking moderate users.

1. Consumptive levels of caffeine significantly increased whole-arm physiological tremor in young adult males at 3 but not 1 mg caffeine/kg bodyweight. 2. Physiological tremor was not affected by time of day (TOD) or the interaction of caffeine and TOD. 3. Findings suggest that ingestion of a single oral dose of caffeine at typical consumptive levels results in measurable changes in physiological functioning and that whole-arm physiological tremor is a sensitive measure of physiological change due to drug effects. 4. However, the results suggest that whole-arm physiological tremor may not detect reliably TOD changes in functioning.

Molecular basis of alpha-thalassemia in Sicily.

To evaluate the allelic frequency and genetic diversity of alpha-thalassemia defects in Sicily, both epidemiological and patient-oriented studies were carried out. For the epidemiological study, phenotypic data were collected on more than 1000 Sicilian individuals. Among them, 427 were explored at the molecular level for nine alpha-thalassemic variants known to be common in the Mediterranean region. Our data reveal an allele frequency of 4.1% for alpha(+)-thalassemia matching that of beta-thalassemia in this region. The presence of alpha0-thalassemia (--MEDI and --CAL) was observed only in the group of referred patients. Newly acquired nucleotide sequence data on the deletional breakpoint of --CAL allowed us to design a simple PCR-based procedure for exploring this allele. The data also provide additional information concerning the genetic mechanisms involved in such large deletions.

Histopathology of thalassemic heart disease: an endomyocardial biopsy study.

Right ventricle endomyocardial biopsies were obtained from 13 thalassemic patients. Clinical profiles were investigated, and serum ferritin tests were assessed using diagnostic kits. Histochemical iron detection (Perls method) and immunohistochemical stain for ferritin were performed in the endomyocardial samples. Histologic iron overload was observed in eight patients, and variable iron deposits were recognized by a semiquantitative method. There was a statistically evident correlation between serum ferritin and myocardial iron storage. Marked iron deposition was associated with higher immunohistologic ferritin concentration. Iron-negative tissue samples showed bland immunohistochemical positivity. Myocardial interstitial fibrosis was observed in 12 cases; diffuse perimyocytic or perivascular fibrosis and endocardium thickening were the main histologic patterns identified. One biopsy was characterized by marked fibrolipomatous infiltration. Myocyte hypertrophy, myocytolysis, and severe capillary congestion also were observed.

Laboratory evaluation of a new delivery system to improve patient compliance with chelation therapy.

As thalassaemia patients live longer, compliance with chelation therapy becomes more and more of a problem. Teenagers rebel against the continual need for treatment, and young adults with jobs, active social lives and families, find it hard to find the time to prepare and deliver their treatments. In an effort to find a solution to this problem, we evaluated a non-electronic, disposable pump (Infusor 5 ml/h, Baxter Healthcare) to determine its suitability for high dose desferrioxamine (DFO) infusions. We wanted an infusion time of 12 h to support overnight delivery in the patient's home. Because of DFOs viscosity, infusion times vary at different concentrations. We found the optimal prescription for total delivery within 12 h to be a dose of 70 mg/kg x patient's body weight diluted in constant volume of 40 ml of water for injection. Our tests were carried out in vitro and the results show that the 5 ml/h Infusor may be used to deliver high dose DFO chelation therapy within 12 h. The variability of the flow rate, dependent on the concentration of the solution, and the absence of a warning system, are more than compensated for by the simplicity of the pump.

High-dose intravenous desferrioxamine (DFO) delivery in four thalassemic patients allergic to subcutaneous DFO administration.

To test the hypothesis that allergy to desferrioxamine is not an immunologic mechanism, but arises from a local effect on the dermal mast cell, we have treated four patients who were not receiving chelation therapy because of hypersensitivity to standard subcutaneous (SC) therapy, with high-dose desferrioxamine (DFO) by the intravenous (IV) route. Three patients had central venous access ports implanted on the anterior chest wall. The fourth patient had the therapy delivered by the peripheral vein route. All patients had the drug delivered via an elastomeric infusor. Intravenous therapy was successful for all patients. During one year of therapy no local or systemic allergic manifestations were noted. In addition, no impairment of hearing or vision or any catheter complications were reported. A very high level of patient compliance to the therapy resulted in dramatically decreased iron stores and ferritin levels (2,759 ng/ml to 717.5 ng/ml) and a significant improvement in the clinical status of all patients. The absence of allergic episodes in this patient group after 1 year of i.v. therapy would strongly support the hypothesis that SC DFO allergy is related to a direct effect on dermal mast cells and is not an immunological reaction. This study suggests that patients with severe allergy to SC DFO can therefore safely receive their chelation therapy via the i.v. route.

Cardiac iron overload in thalassemic patients: an endomyocardial biopsy study.

Secondary heart failure induced by organ siderosis is the main cause of death in patients affected by thalassemia major. At present it cannot be predicted whether heart siderosis is correlated with iron overload and little is known about the real cardiac histological pattern of post transfusional hemochromatosis in patients with thalassemia major and intermedia. The study aim was to evaluate cardiac iron overload by non invasive and invasive techniques. Fifteen thalassemic patients were investigated and endomyocardial biopsy performed in ten revealed different grades of endomyocardial iron overload with histochemical positivity. Non invasive techniques are not able to furnish an exact picture of the cardiac hemochromatosis. There was a significant correlation between serum ferritin and myocardial iron grade. Patients with elevated ferritin levels and poor compliance to chelating therapy are at high risk of severe heart hemochromatosis. It was seen that endomyocardial biopsy is a useful tool in studying myocardial iron.

Caffeine and time of day effects on a force discrimination task in humans.

The effects of caffeine (0 mg/kg, 1 mg/kg, 3 mg/kg) and time of day (TOD) on human performance were studied using a multiple forceband discrimination task (MFDT) and subjective ratings. Self-rated measures of energy level were affected by TOD and caffeine, while mood was affected by TOD. Energy level decreased throughout the day and was offset by caffeine which increased energy level independent of TOD. Self-reported anxiety was not affected by TOD or caffeine. Mood was affected by TOD in a complex cubic trend with late morning and late evening peaks 12 h apart. MFDT performance was affected by TOD, caffeine dosage, and their interaction. Trend analyses showed varying patterns of TOD effects across peak force variability, response latency, response duration, and correct responding. Results support and extend previous findings of TOD influences on the MFDT and support the utility of multicomponent proprioceptive tasks for examining drug effects on performance.

Oral habits and TMJ dysfunction in facial pain and non-pain subjects.

Multifactorial theories for facial pain aetiology have been proposed in the past, although little research has been produced to support a multifactorial aetiology. The present study assessed self-rated oral habits over a 7 day period for facial pain and non-pain subjects with and without a clinical symptom of TMJ dysfunction. The results indicate a significant effect for TMJ dysfunction and teeth clenching and an interaction between TMJ dysfunction and facial pain for biting of the lips and mouth. A significant chi-square test showed that most non-pain controls with TMJ dysfunction reported that they never clenched. The results are discussed in terms of support for a multifactorial aetiology in some facial pain patients and the need for future research to delineate subgroups of facial pain and non-pain subjects.

Radioimmunoscintigraphy with 99mTc-225.28S-F(ab')2 in cutaneous and ocular melanomas: cases of particular clinical interest.

In the attempt to contribute to a correct and early diagnosis of melanoma, this paper critically evaluates, in 40 patients with cutaneous localization and 20 with ocular localization, the results of immunoscintigraphy with 99mTc-225.28S-F(ab')2 and the clinical, instrumental and biopsy findings. While the cutaneous melanoma group is mainly composed of patients subjected to surgical exeresis of the lesion before radioimmunoscintigraphy (RIS), the ocular melanoma group is composed of patients with the primary lesion in situ or previously treated with contact radiotherapy. In the cutaneous melanoma group 5 cases presented falsely positive immunoscintigraphic findings, and only 2 falsely negative. In the ocular melanomas, the percentage of false negatives was higher (n = 7). This is probably attributable to the antigenic expressivity, higher in the metastases than in the primary melanomas. The cases of particular interest are discussed in relation to the clinical picture, to the instrumental examinations and to the histological findings. The work made it possible to contribute to a more correct interpretation of the RIS findings in the staging and follow-up of cutaneous and ocular melanomas.

Time of day effects on a human force discrimination task.

Although numerous studies have demonstrated reliable relationships between various human performance measures and time of day, disagreement exists concerning the shape of these relationships and their dependence on task variables. Most perceptual-motor tasks emphasize responsiveness to exteroceptive stimuli. We used a multiple force-band discrimination task that requires responsiveness to both exteroceptive and proprioceptive information. Results for a response duration measure showed a quadratic time of day trend similar to previously reported performance tasks. Response latency to the force emission cue and number of correct inband force emissions showed cubic time of day trends not typically reported in the time of day and performance literature. These results have implications for time-of-day effects on real world perceptual performance.

Presence of an African beta-globin gene cluster haplotype in normal chromosomes in Sicily.

African admixture in Sicily has been long suspected because of the presence of the sickle gene. Nevertheless, the degree of African admixture cannot be derived from the study of HbS frequency, since this gene was most likely expanded by the selective pressure of malaria, for a long time endemic to the region. We have examined 142 individuals from the Sicilian town of Butera (12% sickle trait) to search for other markers of the globin gene cluster less likely to be selected for by malaria. The TaqI polymorphism in the intervening sequences between the two gamma genes is informative. We have found only two instances of this African marker (TaqI(-)) among 267 normal chromosomes, demonstrating that the admixture occurred at a much lower level than previously thought.