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Given the poor bioavailability of curcumin, its antinociceptive effects are produced after chronic intravenous administration of high doses, while poly (d,l-lactide-co-glycolide)-loaded vesicles (PLGA) can improve drug delivery. This paper investigates the antinociceptive effects of curcumin-loaded PLGA nanovesicles (PLGA-CUR) administered via intravenous (i.v.) or intrathecal (i.t.) routes at low and high doses. The following models of pain were used: formalin test, zymosan-induced hyperalgesia and sciatic nerve ligation inducing neuropathic allodynia and hyperalgesia. PLGA-CUR administered intravenously was able to reduce the response to nociceptive stimuli in the formalin test and hyperalgesia induced by zymosan. Curcumin, instead, was inactive. Low-dose i.t. administration of PLGA-CUR significantly reduced allodynia produced by sciatic nerve ligation, whereas low doses of curcumin did not change the response to nociceptive stimuli. Long-lasting antinociceptive effects were observed when high doses of PLGA-CUR were administered intrathecally. At high doses, i.t. administration of curcumin only exerted rapid and transient antinociceptive effects. Measurement of cytokine and BDNF in the spinal cord of neuropathic mice demonstrate that the antinociceptive effects of PLGA-CUR depend on the reduction in cytokine release and BDNF in the spinal cord. The results demonstrate the effectiveness of PLGA-CUR and suggest that PLGA-CUR nanoformulation might be a new potential drug in the treatment of pain., (Copyright © 2017 Elsevier B.V. All rights reserved.)

A procedure based on ultra-high-pressure liquid chromatography tandem mass spectrometry has been developed for the determination of mescaline, N,N-dimethyltryptamine, psilocin, psilocybin, salvinorin A in hair of consumers of psychedelic vegetal material such peyote or trichocereus cacti, psilocybe mushrooms, Salvia divinorum or psychedelic beverage ayahuasca. After hair washing with methyl alcohol and diethyl ether and subsequent addition of mescaline-d9 and 3,4-methylenedioxypropylamphetamine as internal standards, hair samples were treated with 250μl VMA-T M3 reagent for 1h at 100°C. After cooling, 100μl M3 extract were diluted with 400μl water and a volume of 10μl was injected into chromatographic system. Chromatographic separation was achieved at ambient temperature using a reverse-phase column and a linear gradient elution with two solvents: 0.3% formic acid in acetonitrile and 5mM ammonium formate pH 3. The mass spectrometer was operated in positive ion mode, using multiple reaction monitoring via positive electrospray ionization. The method was linear from the limit of quantification (0.03-0.05ng/mg depending on analyte under investigation) to 10ng/mg hair, with an intra- and inter-assay imprecision and inaccuracy always less than 15% and an analytical recovery between 79.6% and 97.4%, depending on the considered analyte. Using the validated method, mescaline was found in concentration range of 0.08-0.13ng/mg in hair of peyote smokers, 3.2ng salvinorin A per mg hair were determined in hair from a S. divinorum smoker, 5.6ng N,N-dimethyltryptamine per mg hair from an ayahuasca user and finally 0.8ng psilocybin per ng hair of a psilocybe consumer., (Copyright © 2014 Elsevier B.V. All rights reserved.)

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.

Insulin and C-peptide have been proposed as possible biomarkers of human insulin hormone misuse in sport. An extended intra- and inter-laboratory validation of commercially available immunoassays was performed. Enzyme Amplified Sensitivity Immunoassay (EASIA) assays (Human Insulin-EASIA and C-peptide EASIA kits from BioSource) were evaluated for insulin and C-peptide in serum. The intra- and inter-laboratory precision and accuracy values were good for the evaluated assays with maximum imprecision and inaccuracy of 16% and 23%, respectively, obtained just for one day C-peptide assay evaluation. The range of concentrations found in serum samples under investigation was always covered by the calibration curves of the studied immunoassays. However, a 19.7% of the samples felt below the estimated insulin limit of quantification. High concordance between laboratory results was obtained for insulin assay (intraclass correlation coefficient -ICC=0.857), whereas that for C-peptide was lower (ICC=0.539). Evaluated immunoassays were used to measure serum concentrations of insulin and C-peptide in elite athletes of various sport disciplines at different moment of training season, in recreational athletes at baseline conditions and finally in sedentary individuals. Serum insulin was statistically lower both in recreational and elite athletes when compared to sedentary individuals. Among elite athletes, the specific sport affected serum insulin (e.g., weightlifting) and C-peptide (e.g., triathlon) concentrations. Over the training season, a within athletes variability was observed for taekwondo, swimming and weightlifting athletes. Variations due to those aspects should be taken in careful consideration in the hypothesis of setting reference concentration ranges for doping detection.

Since 2002, the Istituto Superiore di Sanità, Rome, Italy, in cooperation with Institut Municipal d'Investigaciò Mèdica, Barcelona, Spain, has set up an external proficiency testing program (HAIRVEQ) to evaluate reliability in hair testing for drug abuse by laboratories from the Italian National Health Service. The results obtained in the last 2 rounds (2004-2005) by 26 laboratories and the evolution of the performance in hair testing for drugs of abuse by laboratories that have participated during the whole external proficiency testing program are presented. The 3 hair samples from the last exercise (2005) were also included in the proficiency test organized by the Society of Hair Testing (SoHT) and 17 international laboratories reported results. Samples analyzed in both exercises were real hair samples from drug consumers. In 2004, 2 identical samples were sent containing cocaine and opiates. One sample was a pulverized specimen and the second one was cut in short segments. In 2005, 2 samples, one containing MDMA and another containing cocaine, were included together with one blank sample. In 2004, approximately 42% of HAIRVEQ laboratories reported an erroneous qualitative result. The scatter of quantitative results was high, although no statistical differences, except for codeine, were found between results reported for the hair specimen if pulverized or reduced in short cuts. In 2005, 47 incorrect qualitative results were reported by HAIRVEQ laboratories, whereas only 5 were informed by SoHT laboratories. Concerning quantitative results, the ones from HAIRVEQ laboratories were comparable, although more dispersed, than those reported by SoHT laboratories. The scatter in quantitative results remained quite high and similar to those of the previous years; nonetheless, an improvement in the qualitative performance was observed. Considering the few number of laboratories showing a satisfying performance, guidelines have to be provided focused on method validation and qualitative and quantitative data evaluation.

The measurement of serum erythropoietin (EPO) has been proposed as one of the indirect biomarkers for the detection of recombinant human EPO misuse in sport. An extended inter-laboratory validation of two commercial immunoassays for EPO measurement is described. A chemiluminescent immunoassay kit (CHEM) and an enzyme-linked immunosorbent assay kit (ELISA) were evaluated. The CHEM assay showed intra-laboratory precision better than 6% and correct accuracy values for all quality control samples tested. Precisions and accuracies better than 7 and 13%, respectively, were obtained for the ELISA assay for most of the quality control samples. The limit of quantification estimated for CHEM assay was lower than for the ELISA assay. Inter-laboratory concordance was good for both the assays, with lower dispersion shown by the CHEM assay. Results obtained with the ELISA assay were always lower than those of the CHEM assay. However, a good inter-technique correlation was obtained ([ELISA]=0.76 [CHEM]+0.06, r2=0.92). Quality control samples had a good stability after one and two freeze/thaw cycles and in simulated transportation conditions. In conclusion, CHEM and ELISA assays showed similar characteristics regarding intra-laboratory validation. Better inter-laboratory results were obtained with the CHEM assay and, hence, it is considered the recommended assay for anti-doping control analysis.

We studied the activity of some enzymes directly involved in the endogenous antioxidative defense system: glutathione-peroxidase (GPX), glutathione s-transferase (GST) and superoxide dismutase (SOD). We have investigated the effects of selenium and vitamin E diet supplementation, in form of selenium-vitamin E enriched yeast, in Wistar rats that were undergone to surgical right nephrectomy and 30 minutes of hypoxia. Blood samples were tested for several parameters as glucose, cholesterol, etc. to assess the general health conditions. The protocol consisted of 3 groups of 25 Wistar rats: a control group, a pre-fed group and a post-fed group. The results showed a significative difference in the behaviour of azotemia, proteins and cholesterol. In the control group the activity rapidly increased, then the values decreased slowly and differently for each substance. The pre and post-fed group showed a pronounced increase after 48 h but the normal values are reached more rapidly. We observed an increase in the activity of the GPX and GST after surgical operation and ischemia, but the GPX in pre-fed group reached the normal value before the other groups.

The influence of uremic middle molecules on in vitro lymphocyte blast transformation and interleukin-2 production was studied in 12 patients on long-term hemodialysis and 12 healthy control subjects with normal kidney function. Middle molecules inhibit phytohemoagglutinin induced lymphocyte proliferation in a concentration dependent fashion, achieving more than 50% inhibition of tritiated thymidine incorporation after 72 hrs in culture. In addition, there was less interleukin-2 production in the long-term hemodialysis patients studied compared with healthy control subjects.

N-Acetyl-beta-glucosaminidase (NAG) and beta-glucuronidase were measured in the serum of 70 patients with breast and digestive-tract neoplasms and in 70 healthy subjects. The mean value of the NAG activity was significantly (P < 0.001) elevated in patients with gastric, liver and pancreas cancer as compared with the reference population. In patients with liver and pancreas cancer the very high sensitivity contrasted with a low specificity. NAG elevations above normal were observed in 14 (78%) patients with breast cancer, in 11 (100%) with gastric cancer, in 17 (70%) with colorectal cancer, in 8 (100%) with liver cancer and in 9 (100%) with pancreas cancer. In patients with breast and gastric cancer the enzyme shows a good specificity and sufficient sensitivity as a tumor marker. beta-Glucuronidase appeared less sensitive and was significantly elevated (100%) only in patients with pancreas cancer.