Analysis of cocaine and nicotine metabolites in wastewater by liquid chromatography-tandem mass spectrometry. Cross abuse index patterns on a major community

“NOTICE: this is the author’s version of a work that was accepted for publication in Science of the Total Environment. Changes resulting from the publishing process, such as peer review, editing, corrections, structural formatting, and other quality control mechanisms may not be reflected in this document. Changes may have been made to this work since it was submitted for publication. A definitive version was subsequently published in Science of the Total Environment, [VOL 487, (July 2014)] DOI 10.1016/j.scitotenv.2013.10.042."A method based on sample preparation by solid phase extraction and analysis by liquid chromatography and mass spectrometry was validated and used for simultaneous analysis of cocaine, benzoylecgonine and cotinine in samples collected at the major wastewater treatment plant in the city of Lisbon. The aim was to estimate the consumption of both cocaine and nicotine in this community and establish an index involving both drugs supported by the relevance of nicotine as a significant anthropogenic marker. The study was made on two different weekdays during a month in order to evaluate patterns of consumption outside weekends. Cocaine and nicotine ingestion levels were back-calculated and expressed as mass of pure drugs consumed per day and per 1000 inhabitants (mean: 0.604 g and 5.860 g respectively). Cocaine was also expressed on the basis of local drug purity levels (33.7%) with a corresponding increase on dose assessments, and community drug abuse profiles. The authors sustain that this approach should always be included in drug studies of this kind allowing a better drug abuse assessment. No significant different patterns of consumption were obtained during the working days studied with the exception of one case coincident with a national holiday that showed an increased typical profile found on other non-working day studies, namely weekends. A fairly significant relationship was found between nicotine and cocaine consumption that should be further evaluated in future studies. Pharmacokinetic considerations were made and proposed for cocaine assessment based on the impact on back calculations after common simultaneous consumption of cocaine and ethanol.

New perspectives on bioactivity of olive oil: evidence from animal models, human interventions and the use of urinary proteomic biomarkers

Olive oil (OO) is the primary source of fat in the Mediterranean diet and has been associated with longevity and a lower incidence of chronic diseases, particularly CHD. Cardioprotective effects of OO consumption have been widely related with improved lipoprotein profile, endothelial function and inflammation, linked to health claims of oleic acid and phenolic content of OO. With CVD being a leading cause of death worldwide, a review of the potential mechanisms underpinning the impact of OO in the prevention of disease is warranted. The current body of evidence relies on mechanistic studies involving animal and cell-based models, epidemiological studies of OO intake and risk factor, small- and large-scale human interventions, and the emerging use of novel biomarker techniques associated with disease risk. Although model systems are important for mechanistic research nutrition, methodologies and experimental designs with strong translational value are still lacking. The present review critically appraises the available evidence to date, with particular focus on emerging novel biomarkers for disease risk assessment. New perspectives on OO research are outlined, especially those with scope to clarify key mechanisms by which OO consumption exerts health benefits. The use of urinary proteomic biomarkers, as highly specific disease biomarkers, is highlighted towards a higher translational approach involving OO in nutritional recommendations

Evaluation of cardiovascular protective effect of different apple varieties - Correlation of response with composition

NOTICE: this is the author’s version of a work that was accepted for publication in Food Chemistry. Changes resulting from the publishing process, such as peer review, editing, corrections, structural formatting, and other quality control mechanisms may not be reflected in this document. Changes may have been made to this work since it was submitted for publication. A definitive version was subsequently published in Food Chemistry, Vol. 135, Issue 4, 2012. DOI http://dx.doi.org/10.1016/j.foodchem.2012.07.067"Epidemiological evidence supports the concept that diets rich in fruits and vegetables promote health and attenuate or delay the onset of cardiovascular disease (CVD). In particular, a reduced risk of CVD has been associated with apple consumption, probably due to the cholesterol-lowering effect of the main bioactive compounds, namely fiber and polyphenols. In this work, the effect of diet supplementation with 20% of three Portuguese apple cultivars (Bravo de Esmolfe, Malápio Serra and Golden), containing distinct phenolic and fiber concentrations, on serum lipid profile and oxLDL of male Wistar rats fed a cholesterol-enriched diet (2%) was evaluated. After 30 days, only Bravo de Esmolfe apple was able to decrease significantly serum levels of triglycerides, total and LDL cholesterol concentrations (reductions of 27.2%, 21.0% and 20.4%, respectively, in relation to the cholesterol-enriched diet group, p<0.05) . The levels of oxLDL were also significantly improved with the consumption of this apple variety (reductions of 20.0% and 11.9%, in relation to the cholesterol-enriched diet group and control group, respectively, p>0.05) as well as with Malapio da Serra apple (reductions of 9.8% in relation to the cholesterol-enriched diet group, p<0.05). Correlation of the bioactive response with chemical composition showed that catechin, epicatechin, procyanidin B1 and -carotene are the major phytocompounds responsible for the cholesterol lowering ability of apples. The antioxidant potential may have also contributed to this beneficial effect.

Phytochemical Profile of Opuntia ficus-indica (L.) Mill Fruits (cv. ‘Orito’) Stored at Different Conditions

This research analyzed the phytochemical profile of prickly pear fruits from ‘Orito’ cultivar stored under cold conditions (2◦ C, 85–90% RH) and shelf-life conditions at room temperature (stored at 20◦ C for three days after cold storage) for 28 days, mimicking the product life cycle. A total of 18 compounds were identified and quantitated through HPLC-DAD-MS/MS (High-Performance Liquid Chromatographic-Diode Array Detector-Mass Spectrometry) analyses. Phenolic acids such as eucomic acid and betalains such as indicaxanthin were the predominant chemical families, and piscidic acid was the most abundant compound. During cold storage, the content of eucomic acid isomer/derivative and syringaresinol increased, and citric acid decreased, which could be caused by the cold activation of the phenylalanine ammonia-lyase (PAL) and polyphenol oxidase (PPO) enzymes. However, no significant differences were found in the content of these compounds during shelf-life storage. These results showed that ‘Orito’ fruit marketability would be possible up to 28 days after harvesting, retaining its profile, which is rich in bioactive compounds

Method development for measurement of elements in Hungarian red wines by inductivelvy coupled plasma optival emission spectrometry (ICP-OES)

ICP-OES (inductively coupled plasma optical emission spectrometry) was used to determine the element content of red wines. Development of the method involves various procedures of sample handling as well as repeatability and reproducibility measurements. Measurements were made for the following 16 elements: Al, B, Ba, Ca, Cd, Co, Cr, Cu, Fe, K, Mg, Mn, Na, P, Sr and Zn. The relative standard deviation of repeatability ranged from 0.41% (potassium) to 27% (cadmium) and that of reproducibility varied between 0.73% (boron) to 52% (cobalt). Recovery of the elements was determined by standard addition with results ranging from 90.6% (phosphorus) to 116.2% (boron). After that, 15 red wine samples were measured. Significant differences were found in the Al, B, Ca, Co, Cr, Cu, Fe, K, Mg, Mn, Na, P, Sr and Zn (P<0.001) concentration of wine samples. Correlation studies revealed parallel changes in phosphorus-potassium, phosphorus-magnesium and zinc-manganese concentrations (r=0.857, 0.875, 0.838). According to principal component analysis, measurements of zinc-, sodium-, boron-, copper- and strontium content gave almost the same results as obtained with 16 different elements. According to a two dimensional non-linear map of absolute values of principal component analysis, wines from Szekszárd and Eger could be differentiated, whereas the analysis of wines from Villány, the Mátra Mountains and Lake Balaton showed overlapping results

do we understand individual variability?

Introduction: Phenolic acids are important phenolic compounds widespread in foods, contributing to nutritional and organoleptic properties. Factors affceting individual variability: The bioavailability of these compounds depends on their free or conjugated presence in food matrices, which is also affected by food processing. Phenolic acids undergo metabolism by the host and residing intestinal microbiota, which causes conjugations and structural modifications of the compounds. Human responses, metabolite profiles and health responses of phenolics, show considerable individual variation, which is affected by absorption, metabolism and genetic variations of subjects. Opinion: A better understanding of the gut-host interplay and microbiome biochemistry is becoming highly relevant in understanding the impact of diet and its constituents. It is common to study metabolism and health benefits separately, with some exceptions; however, it should be preferred that health responders and non-responders are studied in combination with explanatory metabolite profiles and gene variants. This approach could turn interindividual variation from a problem in human research to an asset for research on personalized nutrition.publishersversionpublishe

Atypical Phenotype in Two Patients with LAMA2 Mutations

Congenital muscular dystrophy type 1A is caused by mutations in the LAMA2 gene, which encodes the a2-chain of laminin. We report two patients with partial laminin-a2 deficiency and atypical phenotypes, one with almost exclusive central nervous system involvement (cognitive impairment and refractory epilepsy) and the second with marked cardiac dysfunction, rigid spine syndrome and limb-girdle weakness. Patients underwent clinical, histopathological, imaging and genetic studies. Both cases have two heterozygous LAMA2 variants sharing a potentially pathogenic missense mutation c.2461A>C (p.Thr821Pro) located in exon 18. Brain MRI was instrumental for the diagnosis, since muscular examination and motor achievements were normal in the first patient and there was a severe cardiac involvement in the second. The clinical phenotype of the patients is markedly different which could in part be explained by the different combination of mutations types (two missense versus a missense and a truncating mutation)

The Sight Loss and Vision Priority Setting Partnership (SLV-PSP): overview and results of the research prioritisation survey process

Objectives: The Sight Loss and Vision Priority Setting Partnership aimed to identify research priorities relating to sight loss and vision through consultation with patients, carers and clinicians. These priorities can be used to inform funding bodies’ decisions and enhance the case for additional research funding. Design: Prospective survey with support from the James Lind Alliance. Setting: UK-wide National Health Service (NHS) and non-NHS. Participants: Patients, carers and eye health professionals. Academic researchers were excluded solely from the prioritisation process. The survey was disseminated by patient groups, professional bodies, at conferences and through the media, and was available for completion online, by phone, by post and by alternative formats (Braille and audio). Outcome measure: People were asked to submit the questions about prevention, diagnosis and treatment of sight loss and eye conditions that they most wanted to see answered by research. Returned survey questions were reviewed by a data assessment group. Priorities were established across eye disease categories at final workshops. Results: 2220 people responded generating 4461 submissions. Sixty-five per cent of respondents had sight loss and/or an eye condition. Following initial data analysis, 686 submissions remained which were circulated for interim prioritisation (excluding cataract and ocular cancer questions) to 446 patients/carers and 218 professionals. The remaining 346 questions were discussed at final prioritisation workshops to reach agreement of top questions per category. Conclusions: The exercise engaged a diverse community of stakeholders generating a wide range of conditions and research questions. Top priority questions were established across 12 eye disease categories. This is an Open Access article distributed in accordance with the Creative Commons Attribution Non Commercial (CC BY-NC 3.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited and the use is non-commercial

Intronic long interspersed nuclear element (LINE-1) insertion in the DMD gene as a cause of Becker Muscular Dystrophy

Long interspersed nuclear elements (LINE-1 or L1) arethe most abundant retrotransposable elements accountingfor nearly 17% of the human genome. These elementscan be randomly incorporated in the genome, thereforehaving an important role in its plasticity and in generatingstructural genetic variants. It has been demonstrated thatL1 retrotransposon activity may occasionally cause geneticdiseases. To date, only four disease-causing L1 elementshave been described in the dystrophin (DMD) gene; threeinserted in exons 44, 48 and 67, in patients with a Duchennemuscular dystrophy (DMD) phenotype, and one detectedin the 5Žuntranslated region, in two apparently unrelatedJapanese families with X-linked dilated cardiomyopathy.We report a 48 year old man with a clinical diagnosisof Becker muscular dystrophy (BMD), in 2001, withoutmolecular confirmation by multiplex PCR and Southern-Blotanalysis, and whose diagnosis was recently revisited becausehis daughter is considering pregnancy. A second molecularstudy, resorting to multiplex ligation-probe amplification(MLPA) analysis and genomic DMD gene sequencing, againfailed to detect abnormalities. A new muscle biopsy showeddystrophic features with irregular labeling for dystrophin onimmunohistochemical analysis, suggesting dystrophinopathy.With the intention of unveiling a genetic defect thatmight be refractory to the previous diagnostic techniques,muscle-derived DMD transcripts were sequenced in theirentirety. Results revealed an insertion of 103 nucleotidesbetween exons 51 and 52, which showed no homology to thegenes reference sequence. Extensive bioinformatic analysis(homology search and splice-site/branch-point analysis) andsequential direct sequencing enabled the discovery of a deepintronicinsertion of an L1 element, in intron 51. This extremelyrare mutational event resulted in the partial exonization of theL1 plus 5 nucleotides of intron 51. In addition to the aberrant out-of-frame transcript, a residually expressed wild-typetranscript was also detected, thereby explaining the milderphenotype in this patient.To our knowledge this is the first report ever ofdystrophinopathy caused by an intronically placed L1element. Besides representing an exceptional contributiontowards widening the DMD gene mutation spectrum, thisstudy highlights the importance of conducting mRNA studiesin as yet uncharacterized BMD/DMD patients. This holds trueeven considering some of the most recent state-of-the-artscreening approaches, based on next-generation sequencingtechnology, where this type of mutation may ultimately fail tobe detected