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Purpose: Progressive inherited retinal degenerations (IRDs) affecting rods and cones are clinically and genetically heterogeneous and can lead to blindness with limited therapeutic options. The major gene defects have been identified in subjects of European and Asian descent with only few reports of North African descent., Methods: Genome, targeted next-generation, and Sanger sequencing was applied to cohort of ∼4000 IRDs cases. Expression analyses were performed including Chip-seq database analyses, on human-derived retinal organoids (ROs), retinal pigment epithelium cells, and zebrafish. Variants' pathogenicity was accessed using 3D-modeling and/or ROs., Results: Here, we identified a novel gene defect with three distinct pathogenic variants in UBAP1L in 4 independent autosomal recessive IRD cases from Tunisia. UBAP1L is expressed in the retinal pigment epithelium and retina, specifically in rods and cones, in line with the phenotype. It encodes Ubiquitin-associated protein 1-like, containing a solenoid of overlapping ubiquitin-associated domain, predicted to interact with ubiquitin. In silico and in vitro studies, including 3D-modeling and ROs revealed that the solenoid of overlapping ubiquitin-associated domain is truncated and thus ubiquitin binding most likely abolished secondary to all variants identified herein., Conclusion: Biallelic UBAP1L variants are a novel cause of IRDs, most likely enriched in the North African population., Competing Interests: Conflict of Interest The authors declare no conflicts of interest., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

Abstract: SETBP1 mutations are found in various clonal myeloid disorders. However, it is unclear whether they can initiate leukemia, because SETBP1 mutations typically appear as later events during oncogenesis. To answer this question, we generated a mouse model expressing mutated SETBP1 in hematopoietic tissue: this model showed profound alterations in the differentiation program of hematopoietic progenitors and developed a myeloid neoplasm with megakaryocytic dysplasia, splenomegaly, and bone marrow fibrosis, prompting us to investigate SETBP1 mutations in a cohort of 36 triple-negative primary myelofibrosis (TN-PMF) cases. We identified 2 distinct subgroups, one carrying SETBP1 mutations and the other completely devoid of somatic variants. Clinically, a striking difference in disease aggressiveness was noted, with patients with SETBP1 mutation showing a much worse clinical course. In contrast to myelodysplastic/myeloproliferative neoplasms, in which SETBP1 mutations are mostly found as a late clonal event, single-cell clonal hierarchy reconstruction in 3 patients with TN-PMF from our cohort revealed SETBP1 to be a very early event, suggesting that the phenotype of the different SETBP1+ disorders may be shaped by the opposite hierarchy of the same clonal SETBP1 variants., (© 2024 American Society of Hematology. Published by Elsevier Inc. All rights are reserved, including those for text and data mining, AI training, and similar technologies.)

Background: Infancy is a complex period of human life, in which environmental experiences have a fundamental role for neurodevelopment. Although conditions of social and sensory deprivation are uncommon in high income countries, the Covid-19 pandemic abruptly modified this condition, by depriving people of their social stimuli of daily life., Aim: To understand the impact of this deprivation on infants' behaviour, we investigated the short-term effects of isolation and use of individual protective systems by mothers during the first two weeks of life., Methods: The study included 11 mother-infant dyads with mothers tested positive to SARS-CoV-2 at the time of delivery (Covid group) and 11 dyads with a SARS-CoV-2 negative mother as controls. Neurobehavioral, visual, and sensory processing assessments were performed from birth to 3 months of age., Results: Findings showed the effect of deprivation on some neurobehavioral abilities of infants in the Covid group; in addition, differences in sensory maturation trends were observed, although they tended to gradually decrease until disappearance at 3 months of age., Conclusion: These findings suggest the significant effects of early sensory and social deprivation during the first two weeks of life, but also provide several insights on the ability of the brain to restore its aptitudes by deleting or reducing the effects of early deprivation before the critical periods' closure., Competing Interests: Declaration of competing interest The authors declare no conflict of interest., (Copyright © 2023 The Authors. Published by Elsevier B.V. All rights reserved.)

Acute myeloid leukemia (AML) is a hematological malignancy derived from neoplastic myeloid progenitor cells characterized by abnormal clonal proliferation and differentiation. Although novel therapeutic strategies have recently been introduced, the prognosis of AML is still unsatisfactory. So far, the efficacy of chimeric antigen receptor (CAR)-T-cell therapy in AML has been hampered by several factors, including the poor accumulation of the blood-injected cells in the leukemia bone marrow (BM) niche in which chemotherapy-resistant leukemic stem cells reside. Thus, we hypothesized that overexpression of CXCR4, whose ligand CXCL12 is highly expressed by BM stromal cells within this niche, could improve T-cell homing to the BM and consequently enhance their intimate contact with BM-resident AML cells, facilitating disease eradication. Specifically, we engineered conventional CD33.CAR-cytokine-induced killer cells (CIKs) with the wild-type (wt) CXCR4 and the variant CXCR4R334X, responsible for leukocyte sequestration in the BM of patients with warts, hypogammaglobulinemia, immunodeficiency, and myelokathexis syndrome. Overexpression of both CXCR4wt and CXCR4mut in CD33.CAR-CIKs resulted in significant improvement of chemotaxis toward recombinant CXCL12 or BM stromal cell-conditioned medium, with no observed impairment of cytotoxic potential in vitro. Moreover, CXCR4-overexpressing CD33.CAR-CIKs showed enhanced in vivo BM homing, associated with a prolonged retention for the CXCR4R334X variant. However, only CD33.CAR-CIKs coexpressing CXCR4wt but not CXCR4mut exerted a more sustained in vivo antileukemic activity and extended animal survival, suggesting a noncanonical role for CXCR4 in modulating CAR-CIK functions independent of BM homing. Taken together, these data suggest that arming CAR-CIKs with CXCR4 may represent a promising strategy for increasing their therapeutic potential for AML., (© 2023 by The American Society of Hematology. Licensed under Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International (CC BY-NC-ND 4.0), permitting only noncommercial, nonderivative use with attribution. All other rights reserved.)

Gain-of-function mutations on STIM1 and ORAI1 genes are responsible for an increased store-operated calcium entry, and underlie the characteristic symptoms of three overlapping ultra-rare genetic disorders (i.e tubular aggregate myopathy, Stormorken syndrome, York platelet syndrome) that can be grouped as tubular aggregate myopathies. These mutations lead to a wide spectrum of defects, which usually include muscle weakness and cramps. Negative modulators of store-operated Ca 2+ -entry targeting wild-type STIM1 and ORAI1 have entered clinical trials for a different array of disorders, including pancreatitis, COVID-19, cancer, and autoimmune disorders and, while efficacy data is awaited, safety data indicates tolerability of this STIM1/ORAI1 mutations are amenable to pharmacological intervention. If this were so, given that there are no approved treatments or clinical trials ongoing for these rare disorders, it could be envisaged that these agents could also rehabilitate tubular aggregate myopathy patients. In the present contribution we characterized the Ca 2+ -entry patterns induced by eleven STIM1 and three ORAI1 mutations in heterologous systems or in patient-derived cells, i.e. fibroblasts and myotubes, and evaluated the effect of CIC-37 and CIC-39, two novel store-operated calcium entry modulators. Our data show that all STIM1 and ORAI1 gain-of-function mutations tested, with the possible exception of the R304Q STIM1 mutation, are amenable to inhibition, albeit with slightly different sensitivities, paving the way to the development of SOCE modulators in tubular aggregate myopathies., (Copyright © 2022. Published by Elsevier Ltd.)

Epidemiological evidence has confirmed the potential causal relationship between specific dietary factors and non-communicable diseases. However, currently nutrition was shown to be insufficiently integrated into medical education, regardless of the country. Without an adequate nutrition education, it is reasonable to assume that future physicians, as well as other health care professionals, will be not able to provide the highest quality care to patients in preventing and treating non-communicable diseases. Furthermore, the insufficient availability of physicians with specializations in nutrition has posed the basis for the development of non-medical careers in the field of nutrition. The present document was drafting by the Italian College of Academic Nutritionists, MED-49 (ICAN-49), with the aim to provide an overview on the nutritional competency standards covered by several health care professionals (Physicians Clinical Nutrition Specialists, Clinical Dietitians, Professional Clinical Nutrition Specialists, etc) for the prevention of diseases and/or support of pharmacological therapies. The aim of the ICAN 49 is to suggest a major shift in practice opportunities and roles for many nutritionists, especially for the management of the metabolic diseases, and promote a paradigm change: a clinical and educational leadership role for Physician Clinical Nutrition Specialists in the hospital setting., Competing Interests: Declaration of competing interest The authors have no competing financial interests in relation to the work described., (Copyright © 2021 The Italian Diabetes Society, the Italian Society for the Study of Atherosclerosis, the Italian Society of Human Nutrition and the Department of Clinical Medicine and Surgery, Federico II University. Published by Elsevier B.V. All rights reserved.)

Objective: Abdominal aortic aneurysm (AAA) sac shrinkage after endovascular aortic repair (EVAR) has been regarded as positive marker of EVAR success durability. The purpose of this study was to describe the morphovolumetric changes of the AAA sac during follow-up after elective EVAR and to analyze sac shrinkage-related variables., Methods: This is a single-center, retrospective, observational cohort study from a tertiary referral university hospital. All patients treated with EVAR between January 2013 and December 2018 were identified. Inclusion criteria were elective EVAR for AAA, preoperative computed tomography angiography within 6 months before EVAR and at least one postoperative computed tomography angiography during the follow-up, using a standardized protocol. Aneurysm sac shrinkage was defined as diameter decrease of 1 cm or more, volume shrinkage threshold was identified by a 16% decrease compared with the preoperative value. Primary outcomes were early (≤30 days) and late survival, and freedom from aneurysm-related mortality (ARM), and aortic reintervention., Results: There were 149 of the 325 patients (45.8%) who met the inclusion criteria: 133 (89.3%) were male and 16 (10.7%) female. The mean age was 74 ± 7 years (range, 55-87 years); the median AAA diameter was 56 mm (interquartile range, 50.0-61.2 mm) and the median volume was 138.8 cm 3 (range, 99.0-178.3 cm 3 ). Primary technical success was achieved in 145 patients (97.3%). The in-hospital mortality rate was 1.3%. The median follow-up was 42 months (interquartile range, 22.5-58.0 months). Both AAA diameter and volume decreased (P = .001 and P = .035, respectively) compared with preoperative measurements. Diameter shrinkage was adjudicated in 27 patients (18.1%), volume shrinkage was observed in 42 patients (28.2%). A Cox regression analysis demonstrated an association between the AAA diameter shrinkage and the preoperative diameter (P = .002; hazard ratio, 1.03; 95% confidence interval [CI], 1.011-1.052). The presence of a persistent endoleak predicted the absence of volume shrinkage (P = .001; hazard ratio, 7.75; 95% CI, 2.282-26.291). The estimated freedom from ARM was 97.5 ± 1.0% (95% CI, 93-99) at 12 months, and 96 ± 2% (95% CI, 90-98) at both 36 and 60 months. Aortic reintervention during the follow-up period was necessary in 7 patients (4.7%). ARM was only observed in the group characterized by the concomitant absence of diameter and volume shrinkage., Conclusions: Volumetric analysis showed to have higher sensitivity than the simple two-dimensional measurement of the diameter to study AAA sac changes after EVAR. Although no predictor was found to be associated with AAA volume shrinkage, ARM occurred only in the group of AAAs with the absence of volume shrinkage., (Copyright © 2021 Society for Vascular Surgery. Published by Elsevier Inc. All rights reserved.)

In this review article, the occurrence of harpagide in the plant kingdom and its associated biological activities are presented and detailed for the first time. The presence of harpagide has been reported in several botanical families within Asteridae, and harpagide has been observed to exert a wide number of biological activities such as cytotoxic, anti-inflammatory, and neuroprotective. These results show how harpagide can be recovered from several natural sources for several pharmacological purposes even if there is a lot to still be studied. Nowadays, the interest is related to its presence in phytomedicines. Threfore, these studies are useful to support and validate the large use of several plants in the folklore medicine., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Background: The European guidelines for quality assurance in colorectal cancer (CRC) screening recommend that interval cancer rate be expressed as a proportion of background incidence rate., Aim: To determine the crude and adjusted proportional incidence of interval CRC in an Italian regional two-yearly faecal immunochemical test (FIT) screening programme., Methods: The programme (year of implementation, 2005) is targeted at over 1,000,000 people aged 50-69 years. The test is a one-sample OC-Sensor (Eiken Chemical Co., Tokyo, Japan). The study covered one-third of the regional area. Excerpts of 434,295 eligible negative FIT records dated 2005-2012 from 193,193 subjects were retrieved from the regional CRC screening data warehouse. By 31 December 2013, the cohort accumulated 198,302 man-years and 235,370 woman-years. Interval CRCs were identified by record-linkage with the local population-based cancer registry. Their number was divided by the expected number, estimated with age-period-cohort models, to obtain the proportional incidence., Results: The proportional incidence of interval CRC for men and women was, respectively, 0.06 (95% confidence interval (CI), 0.04-0.09) and 0.17 (95% CI, 0.13-0.23) in the first interval year, and 0.21 (95% CI, 0.16-0.26) and 0.28 (95% CI, 0.22-0.36) in the second year., Conclusions: The results were acceptable and in line with previous studies., (Copyright © 2020 Editrice Gastroenterologica Italiana S.r.l. Published by Elsevier Ltd. All rights reserved.)

Background: Few studies have assessed associations of non-enzymatic antioxidant capacity (NEAC) in the overall diet with all-cause or specific mortality, and their results have been inconsistent., Objectives: The present study investigated the association between dietary NEAC and all-cause or cause-specific mortality., Methods: The study was a large-scale population-based prospective cohort study in Japan consisting of 42,520 men and 50,207 women aged 44-76 y, who had no history of cancer, stroke, ischemic heart disease, or chronic liver disease. We evaluated FFQ-based dietary NEAC with use of published databases in which the NEACs of individual foods were analyzed by ferric reducing antioxidant power (FRAP) and oxygen radical absorbance capacity (ORAC) assays. Dietary NEAC was calculated by multiplying the estimated NEAC with the consumed amount and summing up those values for all foods, and was categorized in quartiles. We identified death and cause of death with use of residential registry and death certificates. HRs and 95% Cls for death from the second survey, which was conducted from April 1995 to December 2014 were estimated with Cox proportional hazards regression analysis., Results: After 1,498,308 person-years of follow-up, 12,978 total deaths occurred. The multivariable-adjusted HRs (95% Cls) for all-cause mortality for the highest compared with the lowest quartile of FRAP and ORAC were 0.85 (0.80, 0.89) and 0.84 (0.79, 0.89), respectively. Dietary NEACs were inversely associated with mortality from cardiovascular disease (CVD), but not from cancer. The multivariable-adjusted HRs (95% Cls) for CVD for the highest compared with the lowest quartile of FRAP and ORAC were 0.83 (0.75, 0.92) and 0.79 (0.70, 0.89), respectively., Conclusions: Higher dietary NEACs from FRAP and ORAC were associated with lower risk of all-cause mortality and mortality from CVD in Japanese adults., (Copyright © American Society for Nutrition 2019.)

In this work, the fifth part of an ongoing phytochemical study on Wollemia nobilis was reported. The attention was now focused on the male reproduction organs of which the content in both primary and secondary metabolites was analyzed. Twenty compounds, belonging to seven different classes of natural compounds, were identified from the ethanolic extract by means of Column Chromatography and NMR and MS Spectroscopy. They all represent new compounds for the studied organ whereas some of them are also new constituents of the genus or even previously undescribed phytochemicals. Their presence was able to display a general overview of these organs from the phytochemical standpoint and to provide more elements in confirmation with the current botanical classification of the species. Moreover, they add a further experimental evidence of the tendency of this species to accumulate different metabolites in different organs. This characteristic as well as the occurrence of several compounds with added value, make this plant a possible candidate for large scale cultivation with extractive purposes to obtain useful phytochemicals for botanicals and pharmaceutical fields. Moreover, they offer the opportunity to develop an additional method of conservation and protection for this endangered and very rare species., (Copyright © 2018 Elsevier B.V. All rights reserved.)

An undescribed labdane-like diterpene with a rare spiro-β-lactone function was identified from the ethanol extract of the male cones of the coniferous tree Wollemia nobilis. This spirolabdadienolide (IUPAC name: syn-ent-8(17),13-labdadien-19,18-olid-15-oic acid methyl ester; trivial name: wollemolide), was isolated by means of traditional and high performance chromatography techniques and structurally elucidated through NMR and MS. In addition, six further known metabolites were evidenced in the extract. Wollemolide, which may be considered an additional chemotaxonomic marker, and 4'-O-methyl-scutellarein, a simple flavonoid, had not been isolated in our previous phytochemical study on the same plant organ. This demonstrates how the molecular pattern of a plant species is in continuous movement and changes with the passing of time according to the climate of the year., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Nowadays, practice of tattooing is very common worldwide and, along with this increasing trend, there is also an increased risk of adverse reactions to tattoo pigments that are well known and described in literature. Previous studies have reported that cutaneous and allergic reactions to a particular pigment can manifest in several ways (allergic contact dermatitis and photo-allergic dermatitis). In this paper, a new high-throughput method is presented, in order to achieve a new system for the quality control on tattoo inks based on chromatographic-spectroscopic approach. The samples, twenty-one tattoo inks and three permanent makeup, comprised the following colors: black inks, yellow, blue, green, white, pink and various shades of red (pigment that gives many allergic responses) were analyzed through the combination of chromatographic and spectroscopic techniques, the HPTLC-Raman. In particular, Raman technique has been chosen because of its high sensitivity towards the inorganic and organic pigments, main constituents of tattoo inks. Moreover, the advantage of this hyphenated technique is to overcome the problem of analysing the complex mixture of tattoo inks, allowing to obtain a Raman spectrum of each single component, isolated by chromatographic separation. This approach aims at developing a powerful instrument to establish the nature of tattoo inks and substances that could be cause adverse reactions in tattooed patients., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Despite benzyne has been known since 1940, this fascinating species has received much attention only over the last decades. The renaissance of interest in aryne chemistry is ascribable to the seminal discovery that arynes can be generated in situ from stable and commercially available precursors under mild and neutral conditions. Thanks to their high reactivity, arynes are key intermediates in countless chemical transformations ranging from cycloadditions to insertions and, among all these approaches, multicomponent reactions (MCRs) are currently standing out in this field. In particular, this short article is focused on MCRs involving the concomitant use of benzyne and isocyanides. Furthermore, the first overview on aryne MCRs triggered by α-isocyanoacetamides is presented., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Ajuga chamaepitys (L.) Schreb, well-known as Camaepitium or Ground Pine, is an annual herb typical of the Mediterranean area accounting several uses in the traditional medicine. In this work we have, analyzed the plant iridoid fraction together with the essential oil composition and study of the plant indumentum. Finally, we assayed the polar extracts and essential oil obtained from the aerial parts for antioxidant activity and cytotoxicity on tumor cells. The analysis of the monoterpene glycosides allowed us to isolate from roots and aerial parts and to structurally elucidate by NMR and MS the following compounds: ajugoside (1), reptoside (2), 8-O-acetylharpagide (3), harpagide (4), 5-O-β-d-glucopyranosyl-harpagide (5), asperulosidic acid (6), deacetyl asperulosidic acid (7) and 5-O-β-d-glucopyranosyl-8-O-acetylharpagide (8), among which 5 and 8 were two new natural products. Chemotaxomic relevance of these constituents was discussed. The chemical analysis of A. chamaepitys essential oil by GC-FID and GC-MS showed ethyl linoleate (13.7%), germacrene D (13.4%), kaurene (8.4%), β-pinene (6.8%), and (E)-phytol (5.3%) as the major volatile components. The micromorphological and histochemical study showed that iridoids and essential oil are mainly produced in the type III capitates and peltate trichomes of leaves and flowers. Biological evaluations of A. chamaepitys polar extracts and essential oil showed that the former were more potent as radical scavengers than the latter. MTT assay revealed that essential oil and ethanolic extracts were moderately cytotoxic on tumor cells with IC50 of 36.88 and 59.24μg/mL on MDA-MB 231 cell line, respectively, and IC50 of 60.48 and 64.12μg/mL on HCT116, respectively., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Background: Consumption of fruit and vegetables has been inversely related to gastric cancer. Two studies found that dietary antioxidant capacity has some role in explaining this association. We investigated the overall antioxidant effect from diet on gastric cancer using three measures of non-enzymatic antioxidant capacity (NEAC)., Methods: We used data from an Italian case-control study including 230 patients with incident, histologically confirmed gastric cancer, and 547 frequency matched controls admitted to the same hospitals for acute non-neoplastic diseases. A reproducible and valid food frequency questionnaire was used to assess subjects' usual diet. NEAC was measured using Italian food composition tables in terms of Trolox equivalent antioxidant capacity (TEAC), Ferric reducing-antioxidant power (FRAP) and Total radical-trapping antioxidant parameter (TRAP). We estimated the odds ratios (OR) of gastric cancer and the corresponding 95% confidence intervals (CI) using conditional logistic regression models including terms for recognized gastric cancer risk factors and total energy intake., Results: NEAC was inversely related with gastric cancer risk with ORs for the highest versus the lowest quintile of 0.54 (95%CI, 0.33-0.88) for TEAC, 0.67 (95%CI, 0.42-1.07) for FRAP and 0.57 (95%CI, 0.36-0.90) for TRAP., Conclusions: A diet rich in antioxidant capacity reduced gastric cancer risk, suggesting a high consumption of fruit and vegetables and a moderate consumption of wine and whole cereals., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

Neonatal bone marrow transplantation (BMT) could offer a novel therapeutic opportunity for genetic disorders by providing sustainable levels of the missing protein at birth, thus preventing tissue damage. We tested this concept in mucopolysaccharidosis type I (MPS IH; Hurler syndrome), a lysosomal storage disorder caused by deficiency of α-l-iduronidase. MPS IH is characterized by a broad spectrum of clinical manifestations, including severe progressive skeletal abnormalities. Although BMT increases the life span of patients with MPS IH, musculoskeletal manifestations are only minimally responsive if the timing of BMT delays, suggesting already irreversible bone damage. In this study, we tested the hypothesis that transplanting normal BM into newborn MPS I mice soon after birth can prevent skeletal dysplasia. We observed that neonatal BMT was effective at restoring α-l-iduronidase activity and clearing elevated glycosaminoglycans in blood and multiple organs. At 37 weeks of age, we observed an almost complete normalization of all bone tissue parameters, using radiographic, microcomputed tomography, biochemical, and histological analyses. Overall, the magnitude of improvements correlated with the extent of hematopoietic engraftment. We conclude that BMT at a very early stage in life markedly reduces signs and symptoms of MPS I before they appear., (© 2015 by The American Society of Hematology.)

Background and Aims: Oxidative processes have been related to atherosclerosis, but there is scanty information on the role of dietary antioxidants in the prevention of acute myocardial infarction (AMI)., Methods and Results: The relationship between non-enzymatic antioxidant capacity (NEAC) and the risk of nonfatal AMI was investigated in a case-control study conducted in Milan, Italy, between 1995 and 2003. Cases were 760 patients below 75 years with a first episode of AMI and controls were 682 patients admitted to hospitals for acute conditions, who completed an interviewer-administered food frequency questionnaire, tested for validity and reproducibility. NEAC (excluding coffee) was measured using Italian food composition tables in terms of ferric reducing-antioxidant power (FRAP), Trolox equivalent antioxidant capacity (TEAC) and total radical-trapping antioxidant parameter (TRAP). The odds ratios (OR) of AMI, and the corresponding 95% confidence intervals (CI), were obtained by multiple logistic regression models including terms for main risk factors of AMI and total energy intake. NEAC was inversely related with the risk of AMI. The ORs for the highest quintile compared with the lowest one were 0.41 (95% CI, 0.27-0.63) for FRAP, 0.42 (95% CI, 0.27-0.65) for TEAC and 0.41 (95% CI, 0.27-0.62) for TRAP, with significant trends in risk. The inverse relationship was apparently stronger in women and in subjects aged ≥ 60 years., Conclusions: Our results support a favorable role of dietary NEAC in the prevention of AMI, and encourage a high consumption of fruit and vegetables and a moderate consumption of wine and whole cereals., (Copyright © 2014 Elsevier B.V. All rights reserved.)

Liquid chromatography coupled with tandem mass spectrometry (HPLC-MS/MS) has become the method of choice for analysis in biological matrices, because of its high specificity and sensitivity. However, it should be taken into account that the presence of matrix components coeluting with analytes might interfere with the ionization process and affect the accuracy and precision of the assay. For this reason, the presence of a "matrix effect" should always be evaluated during method development, above all in complex matrix such as urine. In the present work, a HPLC-MS/MS method was developed for the quantification of urinary iPF2α-III and iPF2α-VI. A careful assessment of matrix effect and an accurate validation were carried out, in order to verify the reliability of quantitative data obtained. Ion suppression, due to the matrix components, was reduced through optimization of both chromatographic method and sample extraction procedure. Urine samples were purified by solid phase extraction (SPE) and the extracts injected into the HPLC-MS/MS system, equipped with a TurboIonSpray ionization source operated in negative ion mode (ESI(-)). Stable isotope-labeled analogues (iPF2α-III-d4 and iPF2α-VI-d4) were used as internal standards, and quantification was performed in multiple reaction monitoring (MRM) mode by monitoring the following mass transitions: m/z 353.4→193.2 for iPF2α-III, m/z 357.2→197.0 for iPF2α-III-d4, m/z 353.4→115.1 for iPF2α-VI, and m/z 357.4→115.1 for iPF2α-VI-d4. The validated assay, applied to the analysis of urinary samples coming from healthy and overweight subjects, resulted suitable for an accurate quantification of iPF2α-III and iPF2α-VI in human urine., (Copyright © 2014 Elsevier B.V. All rights reserved.)

Background Aims: Cord blood (CB) and amniotic fluid (AF) could represent new and attractive mesenchymal stromal cell (MSC) sources, but their potential therapeutic applications are still limited by lack of standardized protocols for isolation and differentiation. In particular, chondrogenic differentiation has never been deeply investigated., Methods: MSCs were obtained from CB and AF samples collected during cesarean sections at term and compared for their biological and differentiation properties, with particular interest in cartilage differentiation, in which quantitative real-time polymerase chain reaction and immunohistochemical analyses were performed to evaluate the expression of type 2 collagen, type 10 collagen, SRY-box9 and aggrecan., Results: We were able to isolate MSCs from 12 of 30 (40%) and 5 of 20 (25%) CB and AF units, respectively. Fluorescence in situ hybridization analysis indicated the fetal origin of isolated MSC strains. Both populations expressed mesenchymal but not endothelial and hematopoietic markers, even though we observed a lower expression of human leukocyte antigen (HLA) I in CB-MSCs. No differences in proliferation rate and cell cycle analysis could be detected. After osteogenic induction, both populations showed matrix mineralization and typical marker expression. Under chondrogenic conditions, pellets derived from CB-MSCs, in contrast with AF-MSCs pellets, were significantly larger, showed cartilage-like morphology and resulted positive for chondrocyte-associated markers, such as type 2 collagen, type 10 collagen, SRY-box9 and aggrecan., Conclusions: Our results show that CB-MSCs and AF-MSCs collected at term differ from each other in their biological and differentiation properties. In particular, only CB-MSCs showed a clear chondrogenic potential and thus could represent an ideal candidate for cartilage-tissue engineering., (Copyright © 2014 International Society for Cellular Therapy. Published by Elsevier Inc. All rights reserved.)

Human bone marrow stromal cells (BMSCs, also known as bone marrow-derived "mesenchymal stem cells") can establish the hematopoietic microenvironment within heterotopic ossicles generated by transplantation at non-skeletal sites. Here we show that non-mineralized cartilage pellets formed by hBMSCs ex vivo generate complete ossicles upon heterotopic transplantation in the absence of exogenous scaffolds. These ossicles display a remarkable degree of architectural fidelity, showing that an exogenous conductive scaffold is not an absolute requirement for bone formation by transplanted BMSCs. Marrow cavities within the ossicles include erythroid, myeloid and granulopoietic lineages, clonogenic hematopoietic progenitors and phenotypic HSCs, indicating that complete stem cell niches and hematopoiesis are established. hBMSCs (CD146(+) adventitial reticular cells) are established in the heterotopic chimeric bone marrow through a unique process of endochondral bone marrow formation, distinct from physiological endochondral bone formation. In this process, chondrocytes remain viable and proliferate within the pellet, are released from cartilage, and convert into bone marrow stromal cells. Once explanted in secondary culture, these cells retain phenotype and properties of skeletal stem cells ("MSCs"), including the ability to form secondary cartilage pellets and secondary ossicles upon serial transplantation. Ex vivo, hBMSCs initially induced to form cartilage pellets can be reestablished in adherent culture and can modulate gene expression between cartilage and stromal cell phenotypes. These data show that so-called "cartilage differentiation" of BMSCs in vitro is a reversible phenomenon, which is actually reverted, in vivo, to the effect of generating stromal cells supporting the homing of hematopoietic stem cells and progenitors., (Copyright © 2014. Published by Elsevier B.V.)

Background and Aims: The intake of antioxidant-rich foods may increase the blood levels of non enzymatic antioxidant capacity (NEAC). NEAC takes into account all antioxidants from food and synergistic effects between them. We examined the effect of a 1-year intervention with Mediterranean diet on plasma NEAC and assessed whether it was related to baseline NEAC levels., Methods and Results: Five hundred sixty-four participants at high cardiovascular risk were randomly selected from the PREDIMED (Prevención con DIeta MEDiterránea) Study, a large 3-arm randomized clinical trial. Blood NEAC levels were measured at baseline and after 1-year of dietary intervention with 1) a Mediterranean diet supplemented with virgin olive oil (MED + VOO); 2) a Mediterranean diet supplemented with nuts (MED + nuts), or 3) a control low-fat diet. Plasma NEAC was analyzed using FRAP (ferric reducing antioxidant potential) and TRAP (total radical-trapping antioxidant parameter) assays. Plasma FRAP levels increased after 1-year of intervention with MED + VOO [72.0 μmol/L (95% CI, 34.2-109.9)] and MED + nuts [48.9 μmol/L (24.3-73.5)], but not after the control low-fat diet [13.9 μmol/L (-11.9 to 39.8)]. Participants in the lowest quartile of plasma FRAP at baseline significantly increased their levels after any intervention, while those in the highest quartile decreased. Similar results occurred with TRAP levels., Conclusions: This study shows that a 1-year of MED diet intervention increases plasma TAC level in subjects at high risk for cardiovascular disease. Moreover, the effectiveness of dietary supplementation with antioxidants may be related to baseline levels of plasma NEAC., (© 2013 Elsevier B.V. All rights reserved.)

Background and Aim: A complex relationship between immune system and metabolic pathway exists and can induce oxidative stress. The objective of this study was to design a new methodology allowing the measurement of oxidative status of leukocytes., Methods and Results: We developed a flow cytometry technique, based on C11-BODIPY 581/591 staining, to evaluate peroxidation in leukocytes. We defined the Peroxidation of Leukocytes Index Ratio (PLIR) as the ratio between the damage after AAPH-induced and PMA-induced peroxidation, using Trolox as standard antioxidant. Sensitivity of the method was assessed by correlating results with plasma antioxidant capacity (TRAP and FRAP), levels of endogenous antioxidants (uric acid and sulfhydryls) and markers of metabolic status (cholesterol, triglycerides, glucose and insulin). PLIR measures the ratio between the resistance to exogenous and endogenous ROS injury, independently from baseline level of oxidation, which was directly correlated with plasma cholesterol on lymphocytes (0.738, p=0.029), monocytes (0.691, p=0.047) and neutrophils (0.690, p=0.047). PLIR of lymphocytes was inversely correlated with uric acid (-0.810, p=0.009) and FRAP (-0.738, p=0.029) levels. On the other hand, PLIR of monocytes was directly correlated with the total scavenger antioxidant capacity attributable to nutritional antioxidants (0.738, p=0.029), calculated as the difference between TRAP and the contribution of uric acid and sulfhydryls to its value., Conclusions: This study reports a feasible and reproducible new flow cytometry assay for assessing the leukocytes redox status. PLIR discriminates between reducing and scavenger activities and is able to appreciate the potentially dangerous effect of uric acid on innate immune response., (Copyright © 2013 Elsevier B.V. All rights reserved.)

We propose a new data analysis approach for reactive oxygen species detection using Dihydrorhodamine 123 in blood monocytes and neutrophils. This approach, based on data transformation using lymphocytes as internal standard, allows to appreciate free radical production by monocytes also without Phorbol 12-myristate 13-acetate (PMA) activation. In addition, this method is sensitive to differences in healthy subjects due to sub-pathological conditions, such as hypercholesterolemia., (Copyright © 2012 Elsevier B.V. All rights reserved.)

Human studies provide evidence for beneficial effects of polyphenol-rich foods on cardiovascular health. The antioxidant activity of polyphenols potentially explains these effects, but is the antioxidant activity a reliable predictor for these effects? An International Life Sciences Institute Europe working group addressed this question and explored the potential of antioxidant claims for polyphenols in relation to cardiovascular health by using the so-called Process for the Assessment of Scientific Support for Claims on Foods project criteria. In this process, analytical aspects of polyphenols, their occurrence in foods, dietary intake, and bioavailability were reviewed. Human studies on polyphenols and cardiovascular health were reviewed together with methods for biomarkers of oxidative damage and total antioxidant capacity (TAC). In retrospective studies, F2-isoprostanes and oxidized LDL, the most reliable biomarkers of lipid peroxidation, and measures for TAC showed the expected differences between cardiovascular disease patients and healthy controls, but prospective studies are lacking, and a causal relationship between these biomarkers and cardiovascular health could not be established. Therefore, the physiological relevance of a potential change in these biomarkers is unclear. We found limited evidence that some types of polyphenol-rich products modify these biomarkers in humans. A direct antioxidant effect of polyphenols in vivo is questionable, however, because concentrations in blood are low compared with other antioxidants and extensive metabolism following ingestion lowers their antioxidant activity. Therefore, the biological relevance of direct antioxidant effects of polyphenols for cardiovascular health could not be established. Overall, although some polyphenol-rich foods exert beneficial effects on some biomarkers of cardiovascular health, there is no evidence that this is caused by improvements in antioxidant function biomarkers (oxidative damage or antioxidant capacity).

Background: To study the impact of the dietary antioxidant quercetin on risk of gastric adenocarcinoma., Patients and Methods: Using data from a large Swedish population-based case-control study of gastric cancer (505 cases and 1116 controls), we studied the association between quercetin and risk of anatomic (cardia/noncardia) and histological (intestinal and diffuse) subtypes of gastric cancer., Results: We found strong inverse associations between quercetin and the risk of noncardia gastric adenocarcinoma, with an adjusted odds ratio (OR) of 0.57 (95% confidence interval 0.40-0.83) for the highest quintile (≥11.9 mg) of daily quercetin intake relative to the lowest quintile of intake (<4 mg quercetin/day), supported by a significant decreasing linear trend (P value < 0.001). Similar findings were observed for the intestinal and diffuse subtype. For cardia cancer, we found a less evident and nonsignificant inverse relationship. The protection of quercetin appeared to be stronger among female smokers, with the OR leveled of at values <0.2 in quintiles 3-5 (>6 mg quercetin/day)., Conclusions: High dietary quercetin intake is inversely related to the risk of noncardia gastric adenocarcinoma, and the protection appears to be particularly strong for women exposed to oxidative stress, such as tobacco smoking.

Ultrasound-guided regional anesthetic techniques have shown some advantages over conventional paresthesia and neurostimulation techniques. We report the case of a neurostimulation-guided continuous interscalene block that would have ended in complication were it not for experience with ultrasound-guided regional anesthesia. Familiarity with ultrasound-guided block techniques permitted assessment of a neck hematoma during interscalene block and ultimately allowed successful peripheral nerve block., (Copyright © 2010 Elsevier Inc. All rights reserved.)

In this study, a comparative investigation was performed of HPLC Ascentis (2.7 microm particles) columns based on fused-core particle technology and Acquity (1.7 microm particles) columns requiring UPLC instruments, in comparison with Chromolith RP-18e columns. The study was carried out on mother and vegetal tinctures of Passiflora incarnata L. on one single or two coupled columns. The fundamental attributions of the chromatographic profiles are evaluated using a chemometric procedure, based on the AutoCovariance Function (ACVF). Different chromatographic systems are compared in terms of their separation parameters, i.e., number of total chemical components (m(tot)), separation efficiency (sigma), peak capacity (n(c)), overlap degree of peaks and peak purity. The obtained results show the improvements achieved by HPLC columns with narrow size particles in terms of total analysis time and chromatographic efficiency: comparable performance are achieved by Ascentis (2.7 microm particle) column and Acquity (1.7 microm particle) column requiring UPLC instruments. The ACVF plot is proposed as a simplified tool describing the chromatographic fingerprint to be used for evaluating and comparing chemical composition of plant extracts by using the parameters D% - relative abundance of the deterministic component - and c(EACF) - similarity index computed on ACVF., (Copyright 2010 Elsevier B.V. All rights reserved.)

Stress proteins such as heat shock proteins (Hsps) are up-regulated in cells in response to various forms of stress, like thermal and oxidative stress and inflammation. Hsps prevent cellular damage and increase immunoregulation by the activation of anti-inflammatory T-cells. Decreased capacity for stress-induced Hsp expression is associated with immune disorders. Thus, therapeutic boosting Hsp expression might restore or enhance cellular stress resistance and immunoregulation. Especially food- or herb-derived phytonutrients may be attractive compounds to restore optimal Hsp expression in response to stress. In the present study, we explored three readout systems to monitor Hsp70 expression in a manner relevant for the immune system and evaluated novel Hsp co-inducers. First, intracellular staining and analysis by flow cytometry was used to detect stress and/or dietary compound induced Hsp70 expression in multiple rodent cell types efficiently. This system was used to screen a panel of food-derived extracts with potent anti-oxidant capacity. This strategy yielded the identity of several new enhancers of stress-induced Hsp70 expression, among them carvacrol, found in thyme and oregano. Second, CD4(+) T-cell hybridomas were generated that specifically recognized an immunodominant Hsp70 peptide. These hybridomas were used to show that carvacrol enhanced Hsp70 levels increased T-cell activation. Third, we generated a DNAJB1-luc-O23 reporter cell line to show that carvacrol increased the transcriptional activation of a heat shock promoter in the presence of arsenite. These assay systems are generally applicable to identify compounds that affect the Hsp level in cells of the immune system.

Background: Cocoa drinks containing flavan-3-ols are associated with many health benefits, and conflicting evidence exists as to whether milk adversely affects the bioavailability of flavan-3-ols., Objective: The objective was to determine the effect of milk on the bioavailability of cocoa flavan-3-ol metabolites., Design: Nine human volunteers followed a low-flavonoid diet for 2 d before drinking 250 mL of a cocoa beverage, made with water or milk, that contained 45 micromol (-)-epicatechin and (-)-catechin. Plasma and urine samples were collected for 24 h, and flavan-3-ol metabolites were analyzed by HPLC with photodiode array and mass spectrometric detection., Results: Milk affected neither gastric emptying nor the transit time through the small intestine. Two flavan-3-ol metabolites were detected in plasma and 4 in urine. Milk had only minor effects on the plasma pharmacokinetics of an (epi)catechin-O-sulfate and had no effect on an O-methyl-(epi)catechin-O-sulfate. However, milk significantly lowered the excretion of 4 urinary flavan-3-ol metabolites from 18.3% to 10.5% of the ingested dose (P = 0.016). Studies that showed protective effects of cocoa and those that showed no effect of milk on bioavailability used products that have a much higher flavan-3-ol content than does the commercial cocoa used in the present study., Conclusions: Most studies of the protective effects of cocoa have used drinks with a very high flavan-3-ol content. Whether similar protective effects are associated with the consumption of many commercial chocolate and cocoa products containing substantially lower amounts of flavan-3-ols, especially when absorption at lower doses is obstructed by milk, remains to be determined.

Plant-based foods (such as fruit and vegetables, wine, nuts, natural vegetable oils, and whole grains) are an important component of traditional diets in Mediterranean regions. A large, consistent body of scientific evidence demonstrates that diets rich in plant foods provide protection against degenerative diseases; however, despite the consensus of the evidence about the health effect of plant foods, it is unclear which components of plant-based foods are protective and what their mechanism of action is. One of the hypotheses postulated to explain the protective effect of plant food, the antioxidant hypothesis, is based on their high content of bioactive molecules. Recent evidence suggests that it is the variegate composition of the plant food, an optimal mixture of different antioxidants endowed with complementary mechanism of action and different redox potential, which is at the basis of their effect on health. The global antioxidant efficiency of complex matrixes can be assessed by measuring their total antioxidant capacity (TAC) representing the result of variables such as redox potentials of the compounds present in the matrix and their cumulative and synergistic interaction. In the last years different databases for TAC of plant foods have been developed. Results suggest that coffee might represent a potential contributor to dietary antioxidant intake. In this contribution after describing the main contributors to dietary TAC for different plant food group, we will discuss the potentiality of coffee as a source of "ready to drink" reducing equivalents.

Background: Total antioxidant capacity (TAC) provides an assessment of antioxidant activity and synergistic interactions of redox molecules in foods and plasma., Objective: We investigated the validity and reproducibility of food-frequency questionnaire (FFQ)-based TAC estimates assessed by oxygen radical absorbance capacity (ORAC), total radical-trapping antioxidant parameters (TRAP), and ferric-reducing antioxidant power (FRAP) food values., Design: Validity and reproducibility were evaluated in 2 random samples from the Swedish Mammography Cohort. Validity was studied by comparing FFQ-based TAC estimates with one measurement of plasma TAC in 108 women (54-73-y-old dietary supplement nonusers). Reproducibility was studied in 300 women (56-75 y old, 50.7% dietary supplement nonusers) who completed 2 FFQs 1 y apart., Results: Fruit and vegetables (mainly apples, pears, oranges, and berries) were the major contributors to FFQ-based ORAC (56.5%), TRAP (41.7%), and FRAP (38.0%) estimates. In the validity study, whole plasma ORAC was correlated (Pearson) with FFQ-based ORAC (r = 0.35), TRAP (r = 0.31), and FRAP (r = 0.28) estimates from fruit and vegetables. Correlations between lipophilic plasma ORAC and FFQ-based ORAC, TRAP, and FRAP estimates from fruit and vegetables were 0.41, 0.31, and 0.28, and correlations with plasma TRAP estimates were 0.31, 0.30, and 0.28, respectively. Hydrophilic plasma ORAC and plasma FRAP values did not correlate with FFQ-based TAC estimates. Reproducibility, assessed by intraclass correlations, was 0.60, 0.61, and 0.61 for FFQ-based ORAC, TRAP, and FRAP estimates, respectively, from fruit and vegetables., Conclusions: FFQ-based TAC values represent valid and reproducible estimates that may be used in nutritional epidemiology to assess antioxidant intake from foods. Further studies in other populations to confirm these results are needed.

Epidemiologic studies have demonstrated an inverse association between consumption of fruits and vegetables and morbidity and mortality from degenerative diseases. The antioxidant content of fruits and vegetables may contribute to the protection they offer from disease. Because plant foods contain many different classes and types of antioxidants, knowledge of their total antioxidant capacity (TAC), which is the cumulative capacity of food components to scavenge free radicals, would be useful for epidemiologic purposes. To accomplish this, a variety of foods commonly consumed in Italy, including 34 vegetables, 30 fruits, 34 beverages and 6 vegetable oils, were analyzed using three different assays, i.e., Trolox equivalent antioxidant capacity (TEAC), total radical-trapping antioxidant parameter (TRAP) and ferric reducing-antioxidant power (FRAP). These assays, based on different chemical mechanisms, were selected to take into account the wide variety and range of action of antioxidant compounds present in actual foods. Among vegetables, spinach had the highest antioxidant capacity in the TEAC and FRAP assays followed by peppers, whereas asparagus had the greatest antioxidant capacity in the TRAP assay. Among fruits, the highest antioxidant activities were found in berries (i.e., blackberry, redcurrant and raspberry) regardless of the assay used. Among beverages, coffee had the greatest TAC, regardless of the method of preparation or analysis, followed by citrus juices, which exhibited the highest value among soft beverages. Finally, of the oils, soybean oil had the highest antioxidant capacity, followed by extra virgin olive oil, whereas peanut oil was less effective. Such data, coupled with an appropriate questionnaire to estimate antioxidant intake, will allow the investigation of the relation between dietary antioxidants and oxidative stress-induced diseases.

Aim: This review is intended to be a critical analysis of selected red-ox methodologies aimed at understanding why the biomarker might be valuable and how it can be measured, highlighting any pitfalls and sample interferences, and describing specific cases of application., Data Summary: Over recent years, considerable efforts have been made to identify and validate the markers of the oxidative/antioxidant status of the human body., Conclusion: Different assays must be applied simultaneously and the related information merged in order to obtain a clearer picture of red-ox status. The relevance of the markers needs to be understood in both pathological and physiological terms: more information is needed about how these markers are regulated in human beings and how they are associated with the diet, because improving health and well-being is as important as preventing the risks of disease itself.

Objective. This article presents an overview of the use of intrathecal bupivacaine (with and without opioid), focusing on laboratory data and clinical use for chronic pain. Some background on epidural use is included to support the intrathecal literature. Materials and Methods. Currently available literature (MEDLINE) regarding the use of intrathecal bupivacaine is reviewed. Prior to presenting the intrathecal bupivacaine data, an overview of data related to bupivacaine stability, microbiology, preclinical toxicology, and pharmacokinetics is presented, along with a brief review of the epidural bupivacaine literature. Results. Based on the current available literature, intrathecal bupivacaine appears to be a safe and acceptable method of treatment for chronic pain in both cancer and noncancer patients. The stability and bacteriologic studies support the use of bupivacaine in external or implantable drug administration devices. Toxicity studies in laboratory animals suggest complications only at plasma levels that would not be seen at clinically relevant doses of intrathecal administration. Bupivacaine is a clinically effective addition to intrathecal opioids. Bupivacaine administration is more effective intrathecally, providing better pain relief than epidural administration. Reports of complications are infrequent. Further studies are needed to define the use of intrathecal bupivacaine and should include long-term safety. Compatibility studies will also be needed when bupivacaine is used in combination with other agents. In addition, outcome studies are needed specifically to differentiate use of intrathecal bupivacaine based on the source and mechanism of pain. Conclusions. While there are few long-term randomized prospective studies at this point, we conclude that intrathecal bupivacaine appears to be a safe and efficacious treatment in both cancer and noncancer pain.

There is mounting evidence emphasizing the importance of intracellular antioxidant levels for maintenance of the immune function. The flavonoid quercetin, a natural antioxidant, has been shown to modulate enzymes involved in the regulation of the inflammatory response. However, up to now, there have been no studies describing quercetin levels in cells of the immune system. A gradient reversed-phase HPLC technique to identify and quantify intracellular levels of quercetin and its application in mice splenocytes are described. Mobile phases were a 0.01 M sodium phosphate monobasic solution adjusted to pH 2.8 with 85% orthophosphoric acid (buffer, Solvent A) and methanol (Solvent B) with a flow rate of 1 ml/min. An eight-channel coulometric electrode array detector was used. In vitro supplementation with increasing concentration of quercetin (25, 50, and 100 microM) raises intracellular quercetin levels in a dose-dependent manner. The method has the required features of specificity and sensitivity for monitoring quercetin uptake in cells of the immune system., (Copyright 2000 Academic Press.)

Consensus guidelines developed by an expert panel are helpful to clinicians when there is variation in practice and lack of a firm evidence base for an intervention, such as intraspinal therapy for pain. An internet-based survey of practitioners revealed remarkable variation in practice patterns surrounding intraspinal therapy. This prompted an interdisciplinary panel with extensive clinical experience in intraspinal infusion therapy to evaluate the results of the survey, the systematic reviews of the literature pertaining to this approach, and their own clinical experience with long-term spinal infusions. The panel proposed a scheme for the selection of drugs and doses for intraspinal therapy, and suggested guidelines for administration that would increase the likelihood of a successful outcome. These expert panel guidelines were designed to provide an initial structure for clinical decision making that is based on the best available evidence and the perspectives of experienced clinicians.

Evidence-based medicine depends on the existence of controlled clinical trials that establish the safety and efficacy of specific therapeutic techniques. Many interventions in clinical practice have achieved widespread acceptance despite little evidence to support them in the scientific literature; the critical appraisal of these interventions based on accumulating experience is a goal of medicine. To clarify the current state of knowledge concerning the use of various drugs for intraspinal infusion in pain management, an expert panel conducted a thorough review of the published literature. The exhaustive review included 5 different groups of compounds, with morphine and bupivacaine yielding the most citations in the literature. The need for additional large published controlled studies was highlighted by this review, especially for promising agents that have been shown to be safe and efficacious in recent clinical studies.

Background: The poor accuracy of the clinical examination of pigmented skin lesions (PSLs) is a major limitation of secondary prevention strategies for cutaneous melanoma (CMM). In the last few years, the epiluminescence microscopy (ELM) technique has been used increasingly as an adjunct to clinical examination in the dermatology practice. Although the question of training has emerged as a priority, the diffusion, the effects, and the correlates of educational programmes in ELM have seldom been studied., Methods: Thirty ELM images of PSLs (11 CMMs, 14 melanocytic nevi (MN), and 5 nonmelanocytic lesions (NMLs) each matched with the corresponding clinical or plain photographic image were independently diagnosed before and after a one-day workshop by 83 Italian dermatologists participating in a nationwide educational programme on ELM. The original histology diagnosis was assumed as a gold standard. The overall effect of training on a set of accuracy measures by PSL type was evaluated. The association of the professional sector (public/private), number of years of general experience in dermatology (1-10/>10), average weekly number of PSLs seen (< or =10/11-20/>20), routine use of ELM (no/yes), and area of residence (northern/southern Italy) with the mean number of PSLs correctly diagnosed before and after training was evaluated with the general factorial analysis of variance. The factors associated with improvement between the two tests were evaluated with the analysis of variance for repeated measures., Results: Compared with pretraining data, the average percentage of exact diagnosis increased significantly for all PSLs (CMMs, 72% vs 55%; MN, 68% vs 64%; NMLs, 67% vs 58%; total lesions combined, 69% vs 60%). Baseline as well as final accuracy were independent from the professional sector and the years of experience but were greater among those subjects who reported >20 PSLs per week compared with the reference group (< or =10 PSLs). The routine use of ELM was associated with a slight advantage in pretraining accuracy. The area of residence was the strongest determinant of baseline as well as final accuracy. The effect of training was independent from all factors studied with the exception of the area of residence with a 13% increase in the frequency of exact diagnosis in northern Italy (from 66-79%) and 6% in southern Italy (from 55-61%)., Conclusions: Though insufficient in absolute terms, a measurable increase in ELM accuracy can be achieved even with intense training sessions of short duration. Medical education to ELM in southern Italy should be a priority.

Moderate wine consumption is reputed to exert a protective effect against coronary heart disease (CHD). The nature of the protective compounds is unclear and the mechanisms are incompletely understood. We studied whether the nonalcoholic component of wine increases plasma antioxidant capacity measured as total radical-trapping antioxidant parameter (TRAP), and whether such an effect is associated with the presence of phenolic compounds in plasma. The TRAP and plasma levels of phenolic compounds were measured in 10 healthy subjects after the ingestion of 113 mL of tap water (control) and alcohol-free red and white wine at 1-wk intervals. Both alcohol-free wines possessed an in vitro dose-dependent peroxyl-radical activity, but red wine, with a polyphenol concentration of 363 +/- 48.0 mg/L quercetin equivalent (QE), was 20 times more active (40.0 +/- 0.1 mmol/L) than white wine (1.9 +/- 0.1 mmol/L), which has a polyphenol concentration of 31 +/- 1 mg QE/L. The ingestion of alcohol-free red wine caused significant increases in plasma TRAP values and polyphenol concentrations 50 min after ingestion. Alcohol-free white wine and water had no effects on either of the plasma values. The parallel and prompt increase of antioxidant status and of circulating levels of polyphenols in fasting subjects after bolus ingestion of a moderate amount of alcohol-free red wine suggests that polyphenols are absorbed in the upper gastrointestinal tract and might be directly involved in the in vivo antioxidant defenses.