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  • image/svg+xml art designer at PLoS, modified by Wikipedia users Nina, Beao, JakobVoss, and AnonMoos Open Access logo, converted into svg, designed by PLoS. This version with transparent background. http://commons.wikimedia.org/wiki/File:Open_Access_logo_PLoS_white.svg art designer at PLoS, modified by Wikipedia users Nina, Beao, JakobVoss, and AnonMoos http://www.plos.org/
    Authors: Fouad M.F. Elshaghabee; Fouad M.F. Elshaghabee; Wilhelm eBockelmann; Diana eMeske; +4 Authors

    Pour obtenir un aperçu spécifique des rôles que les micro-organismes pourraient jouer dans la stéatose hépatique non alcoolique (NAFLD), certaines bactéries intestinales et lactiques et une levure (Anaerostipes caccae, Bacteroides thetaiotaomicron, Bifidobacterium longum, Enterococcus fecalis, Escherichia coli, Lactobacillus acidophilus, Lactobacillus fermentum, Lactobacillus plantarum, Weissella confusa, Saccharomyces cerevisiae) ont été caractérisées par une chromatographie liquide haute performance pour la production d'éthanol lorsqu'elles sont cultivées sur différents glucides : hexoses (glucose et fructose), pentoses (arabinose et ribose), disaccharides (lactose et lactulose) et inuline. Les quantités les plus élevées d'éthanol ont été produites par S. cerevisiae, L. fermentum et W. confusa sur le glucose et par S. cerevisiae et W. confusa sur le fructose. En raison de la mannitol-déshydrogénase exprimée dans L. fermentum, la production d'éthanol sur le fructose a été significativement réduite (P < 0,05). Le pyruvate et le citrate, deux accepteurs d'électrons potentiels pour la régénération du NAD+/NADP+, ont considérablement réduit la production d'éthanol avec de l'acétate produit à la place dans L. fermentum cultivé sur glucose et W. confusa cultivé sur glucose et fructose, respectivement. Dans les boues fécales préparées à partir des matières fécales de quatre volontaires en surpoids, on a constaté que l'éthanol était produit lors de l'ajout de fructose. L'ajout d'A. caccae, L. acidophilus, L. fermentum, ainsi que de citrate et de pyruvate, respectivement, a aboli la production d'éthanol. Cependant, l'ajout de W. confusa a entraîné une augmentation significative (P < 0,05) de la production d'éthanol. Ces résultats indiquent que des micro-organismes comme W. confusa, une bactérie lactique hétéro-fermentaire, négative à la mannitol-déshydrogénase, peuvent favoriser la NAFLD par l'éthanol produit à partir de la fermentation du sucre, tandis que d'autres bactéries intestinales et des bactéries lactiques homo- et hétéro-fermentaires mais positives à la mannitol-déshydrogénase peuvent ne pas favoriser la NAFLD. En outre, nos études indiquent que les facteurs alimentaires interférant avec le microbiote gastro-intestinal et le métabolisme microbien peuvent être importants dans la prévention ou la promotion de la NAFLD. Para obtener información específica sobre los roles que podrían desempeñar los microorganismos en la enfermedad del hígado graso no alcohólico (NAFLD, por sus siglas en inglés), algunas bacterias intestinales y del ácido láctico y una levadura (Anaerostipes caccae, Bacteroides thetaiotaomicron, Bifidobacterium longum, Enterococcus fecalis, Escherichia coli, Lactobacillus acidophilus, Lactobacillus fermentum, Lactobacillus plantarum, Weissella confusa, Saccharomyces cerevisiae) se caracterizaron por cromatografía líquida de alto rendimiento para la producción de etanol cuando se cultivaron en diferentes carbohidratos: hexosas (glucosa y fructosa), pentosas (arabinosa y ribosa), disacáridos (lactosa y lactulosa) e inulina. Las cantidades más altas de etanol fueron producidas por S. cerevisiae, L. fermentum y W. confusa en glucosa y por S. cerevisiae y W. confusa en fructosa. Debido a la manitol-deshidrogenasa expresada en L. fermentum, la producción de etanol en fructosa se redujo significativamente (P < 0.05). El piruvato y el citrato, dos aceptores de electrones potenciales para la regeneración de NAD+/NADP+, redujeron drásticamente la producción de etanol con acetato producido en su lugar en L. fermentum cultivado en glucosa y W. confusa cultivado en glucosa y fructosa, respectivamente. En suspensiones fecales preparadas a partir de heces de cuatro voluntarios con sobrepeso, se encontró que el etanol se producía tras la adición de fructosa. La adición de A. caccae, L. acidophilus, L. fermentum, así como citrato y piruvato, respectivamente, abolió la producción de etanol. Sin embargo, la adición de W. confusa resultó en un aumento significativo (P < 0.05) de la producción de etanol. Estos resultados indican que microorganismos como W. confusa, una bacteria de ácido láctico hetero-fermentativa, negativa para manitol-deshidrogenasa, pueden promover NAFLD a través del etanol producido a partir de la fermentación de azúcar, mientras que otras bacterias intestinales y bacterias de ácido láctico homo- y hetero-fermentativas pero positivas para manitol-deshidrogenasa pueden no promover NAFLD. Además, nuestros estudios indican que los factores dietéticos que interfieren con la microbiota gastrointestinal y el metabolismo microbiano pueden ser importantes para prevenir o promover la EHGNA. To gain some specific insight into the roles microorganisms might play in non-alcoholic fatty liver disease (NAFLD), some intestinal and lactic acid bacteria and one yeast (Anaerostipes caccae, Bacteroides thetaiotaomicron, Bifidobacterium longum, Enterococcus fecalis, Escherichia coli, Lactobacillus acidophilus, Lactobacillus fermentum, Lactobacillus plantarum, Weissella confusa, Saccharomyces cerevisiae) were characterized by high performance liquid chromatography for production of ethanol when grown on different carbohydrates: hexoses (glucose and fructose), pentoses (arabinose and ribose), disaccharides (lactose and lactulose), and inulin. Highest amounts of ethanol were produced by S. cerevisiae, L. fermentum and W. confusa on glucose and by S. cerevisiae and W. confusa on fructose. Due to mannitol-dehydrogenase expressed in L. fermentum, ethanol production on fructose was significantly (P < 0.05) reduced. Pyruvate and citrate, two potential electron acceptors for regeneration of NAD+/NADP+, drastically reduced ethanol production with acetate produced instead in L. fermentum grown on glucose and W. confusa grown on glucose and fructose, respectively. In fecal slurries prepared from feces of four overweight volunteers, ethanol was found to be produced upon addition of fructose. Addition of A. caccae, L. acidophilus, L. fermentum, as well as citrate and pyruvate, respectively, abolished ethanol production. However, addition of W. confusa resulted in significantly (P < 0.05) increased production of ethanol. These results indicate that microorganisms like W. confusa, a hetero-fermentative, mannitol-dehydrogenase negative lactic acid bacterium, may promote NAFLD through ethanol produced from sugar fermentation, while other intestinal bacteria and homo- and hetero-fermentative but mannitol-dehydrogenase positive lactic acid bacteria may not promote NAFLD. Also, our studies indicate that dietary factors interfering with gastrointestinal microbiota and microbial metabolism may be important in preventing or promoting NAFLD. لاكتساب بعض الأفكار المحددة حول الأدوار التي قد تلعبها الكائنات الحية الدقيقة في مرض الكبد الدهني غير الكحولي (NAFLD)، تميزت بعض بكتيريا حمض الأمعاء واللاكتيك وخميرة واحدة (Anaerostipes caccae، Bacteroides thetaiotaomicron، Bifidobacterium longum، Enterococcus fecalis، Escherichia coli، Lactobacillus acidophilus، Lactobacillus fermentum، Lactobacillus plantarum، Weissella confusa، Saccharomyces cerevisiae) بتصوير سائل عالي الأداء لإنتاج الإيثانول عند زراعته على كربوهيدرات مختلفة: hexoses (الجلوكوز والفركتوز)، pentoses (الأرابينوز والريبوز)، disaccharides (اللاكتوز واللاكتولوز)، و inulin. تم إنتاج أعلى كميات من الإيثانول بواسطة S. cerevisiae و L. fermentum و W. confusa على الجلوكوز و S. cerevisiae و W. confusa على الفركتوز. بسبب نازعة هيدروجين المانيتول المعبر عنها في L. fermentum، انخفض إنتاج الإيثانول على الفركتوز بشكل كبير (P < 0.05). قلل البيروفات والسيترات، وهما مستقبلان محتملان للإلكترون لتجديد NAD +/NADP+، بشكل كبير من إنتاج الإيثانول مع الأسيتات المنتجة بدلاً من ذلك في L. fermentum المزروع على الجلوكوز و W. confusa المزروع على الجلوكوز والفركتوز، على التوالي. في الملاط البرازي الذي تم تحضيره من براز أربعة متطوعين يعانون من زيادة الوزن، وجد أن الإيثانول يتم إنتاجه عند إضافة الفركتوز. إضافة A. caccae، L. acidophilus، L. fermentum، وكذلك السترات والبيروفات، على التوالي، ألغت إنتاج الإيثانول. ومع ذلك، أدت إضافة W. confusa إلى زيادة كبيرة في إنتاج الإيثانول (P < 0.05). تشير هذه النتائج إلى أن الكائنات الحية الدقيقة مثل W. confusa، وهي بكتيريا حمض اللاكتيك السلبية غير المتجانسة، قد تعزز NAFLD من خلال الإيثانول المنتج من تخمير السكر، في حين أن البكتيريا المعوية الأخرى وبكتيريا حمض اللاكتيك الإيجابية غير المتجانسة ولكن غير المتجانسة قد لا تعزز NAFLD. أيضًا، تشير دراساتنا إلى أن العوامل الغذائية التي تتداخل مع الكائنات الحية الدقيقة في الجهاز الهضمي والتمثيل الغذائي الميكروبي قد تكون مهمة في منع أو تعزيز NAFLD.

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    Frontiers in Microbiology
    Article . 2016 . Peer-reviewed
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    Frontiers in Microbiology
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    Frontiers in Microbiology
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      Frontiers in Microbiology
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    Authors: David Caldevilla-Domínguez; Almudena Barrientos-Báez; Graciela Padilla-Castillo;

    The social confinement resulting from the COVID-19 crisis temporarily reduced greenhouse gas emissions. Although experts contend that the decrease in pollution rates was not drastic, some surveys detect growth in social concern about the climate. In this new climate-conscious environment, municipalities and local governments are promoting a new way of living and caring for cities, even before they can regain national and international freedom of movement. This work analyzes the connections between new climate awareness arising from the COVID-19 crisis, proposals of sustainable citizenship around the world, and its communication on Twitter to educate the new eco-conscious audience. The methodology mixes quantitative and qualitative analysis, using the Twitonomy Premium tool and the Twitter research tool with data extracted at the end of December 2020. Among the top ten most influential and active accounts, the results show educational institutions, local institutions, companies, neighborhoods, associations, and influencers. The impossibility of living in the city has not prevented citizen education and commitment to make real change for when that city and its citizens return to normality. However, this new normality must be different: more ecological, more responsible, more sustainable, and practiced from early childhood.

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    Sustainability
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    Authors: Jorge Montesinos; Anabel Gil; Consuelo Guerri;

    BackgroundWe previously showed that, by activating innate immune receptors Toll‐like 4 (TLR4), adolescent intermittent ethanol (EtOH) exposure causes neuroinflammation, myelin damage, and behavioral dysfunctions. Recent findings reveal that clinically used opioid antagonists naltrexone (NT) and naloxone (NX) inhibit opioid‐induced TLR4 signaling and that NT, NX, and nalmefene (NF), the 6‐methylene derivative of NX, are able to reduce alcohol drinking escalation.MethodsNF (0.1 mg/kg, intraperitoneally) was injected 1 hour prior to EtOH (3 g/kg, intraperitoneally) following intermittent treatment in female (PND35) adolescent mice. Inflammatory molecules, myelin proteins, and apoptotic markers were assessed in the prefrontal cortex (PFC) and striatum/nucleus accumbens (STR/NAcc). The effect of NF on alcohol drinking preference was evaluated in both the wild‐type and TLR4 knockout (KO) adolescent mice. Using astroglial cells, the inhibitory potential of NT, NX, and NF on lipopolysaccharide (LPS), or the EtOH‐triggered TLR4 response, was compared.ResultsOur findings indicate that NF prevents the up‐regulation of cytokines (IL‐1β, IL‐17A, TNF‐α), chemokines (MCP‐1, MIP‐1, KC), and pro‐inflammatory mediators (iNOS, COX‐2), along with myelin damage and apoptotic events, in both PFC and STR/NAcc. NF also abolishes EtOH‐induced escalation of alcohol preference/consumption, but has no effect when administered to TLR4‐KO mice. In vitro experiments indicate that NX and NF inhibit TLR4 activation upon LPS or EtOH stimulation. Immunofluorescence studies and lipid rafts isolation show that NF is able to prevent TLR4 translocation to lipid rafts/caveolae in astrocytes.ConclusionsThese results suggest that NF prevents neuroinflammation and brain damage by blocking the TLR4 response and also support the role of central pro‐inflammatory immune signaling in the modulation of alcohol consumption/addiction.

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    Alcoholism Clinical and Experimental Research
    Article . 2017 . Peer-reviewed
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      Alcoholism Clinical and Experimental Research
      Article . 2017 . Peer-reviewed
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    Authors: Marisa Silva; Vijaya Pratheepa; Luis Botana; Vitor Vasconcelos;

    Harmful Algal Blooms (HAB) are complex to manage due to their intermittent nature and their severe impact on the economy and human health. The conditions which promote HAB have not yet been fully explained, though climate change and anthropogenic intervention are pointed as significant factors. The rise of water temperature, the opening of new sea canals and the introduction of ship ballast waters all contribute to the dispersion and establishment of toxin-producing invasive species that promote the settling of emergent toxins in the food-chain. Tetrodotoxin, ciguatoxin, palytoxin and cyclic imines are commonly reported in warm waters but have also caused poisoning incidents in temperate zones. There is evidence that monitoring for these toxins exclusively in bivalves is simplistic and underestimates the risk to public health, since new vectors have been reported for these toxins and as well for regulated toxins such as PSTs and DSTs. In order to avoid public health impacts, there is a need for adequate monitoring programs, a need for establishing appropriate legislation, and a need for optimizing effective methods of analysis. In this review, we will compile evidence concerning emergent marine toxins and provide data that may indicate the need to restructure the current monitoring programs of HAB.

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    Toxins
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    Toxins
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    Toxins
    Article . 2016
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      Article . 2016
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    Authors: Román, Juan; Colell Riera, Anna; Blasco, Carmen; Caballería, Juan; +3 Authors

    The oxidative metabolism of ethanol by the cytochrome P450 2E1 (CYP2E1) has been recognized to contribute to the ethanol-induced deleterious effects through the induction of oxidative stress. This study compared the effect of ethanol and acetaldehyde in the induction of oxidative stress and activation of transcription factors nuclear factor-κB (NF-κB) and activating protein 1 (AP-1) in HepG2 cells, which do not express CYP2E1, and HepG2 cells transfected with CYP2E1 (E47 cells). Neither ethanol (80 mmol/L) nor acetaldehyde (25-200 μmol/L) caused oxidative stress in HepG2 cells, an effect that was independent of blocking reduced glutathione (GSH) synthesis with buthionine-l -sulfoximine (BSO). However, BSO preincubation caused an overproduction of peroxides and activation of NF-κB and AP-1 in E47 cells even in the absence of ethanol. Furthermore, the incubation of E47 cells with ethanol (80 mmol/L for up to 5 days) depleted cellular GSH stores in both cytosol and mitochondria, reflecting the induction of oxidative stress. Ethanol activated NF-κB and AP-1 in E47 cells, an effect that was prevented by 4-methylpyrazole, potentiated by cyanamide, and attenuated by trolox C. Interestingly, however, despite the inability of acetaldehyde to induce oxidative stress in HepG2, acetaldehyde activated NF-κB and AP-1; in contrast, ethanol failed to activate these transcription factors in HepG2. Thus, our findings indicate that activation of NF-κB and AP-1 by ethanol and acetaldehyde occurs through distinct mechanisms. CYP2E1 is indispensable in the induction of oxidative stress from ethanol, whereas the activation of NF-κB and AP-1 by acetaldehyde is independent of oxidative stress.

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    Hepatology
    Article . 1999 . Peer-reviewed
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    Hepatology
    Article . 1999
    Hepatology
    Article . 1999 . Peer-reviewed
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      Hepatology
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    Authors: Birte Sievers; Jörg Hausdorf; Volkmar Jansson; Susanne Mayer-Wagner; +5 Authors

    AbstractThe aim of this study was to determine if extracorporeal shock wave therapy (ESWT) in vivo affects the structural integrity of articular cartilage. A single bout of ESWT (1500 shock waves of 0.5 mJ/mm2) was applied to femoral heads of 18 adult Sprague–Dawley rats. Two sham‐treated animals served as controls. Cartilage of each femoral head was harvested at 1, 4, or 10 weeks after ESWT (n = 6 per treatment group) and scored on safranin‐O‐stained sections. Expression of tenascin‐C and chitinase 3‐like protein 1 (Chi3L1) was analyzed by immunohistochemistry. Quantitative real‐time polymerase chain reaction (PCR) was used to examine collagen (II)α1 (COL2A1) expression and chondrocyte morphology was investigated by transmission electron microscopy no changes in Mankin scores were observed after ESWT. Positive immunostaining for tenascin‐C and Chi3L1 was found up to 10 weeks after ESWT in experimental but not in control cartilage. COL2A1 mRNA was increased in samples 1 and 4 weeks after ESWT. Alterations found on the ultrastructural level showed expansion of the rough‐surfaced endoplasmatic reticulum, detachment of the cell membrane and necrotic chondrocytes. Extracorporeal shock waves caused alterations of hyaline cartilage on a molecular and ultrastructural level that were distinctly different from control. Similar changes were described before in the very early phase of osteoarthritis (OA). High‐energy ESWT might therefore cause degenerative changes in hyaline cartilage as they are found in initial OA. © 2010 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 28:1050–1056, 2010

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    Journal of Orthopaedic Research®
    Article . 2010 . Peer-reviewed
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      Journal of Orthopaedic Research®
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    Authors: Dominik Popowski; Karolina A. Pawłowska; Melanie Deipenbrock; Andreas Hensel; +4 Authors

    Ethnopharmacological relevance Phaseaoli pericarpium (bean pods) is a pharmacopeial plant material traditionally used as a diuretic and antidiabetic agents. Diuretic activity of pod extracts was reported first in 1608. Since then Phaseoli pericarpium tea figures in many textbooks as medicinal plant material used by patients. Aim of the study Despite the traditional use of extracts from Phaseolium vulgaris pericarp, limited information is available on bioactivity, chemical composition, and bioavailability of such preparations. The following study aimed to investigate the phytochemical composition, the in vitro permeability of selected extract's constituents over the Caco-2 permeation system, and potential antivirulence activity against uropathogenic Escherichia coli of a hydroalcoholic Phaseoli pericarpium extract (PPX) in vitro to support its traditional use as a remedy used in urinary tract infections. Material and methods The chemical composition of the extract PPX [ethanol:water 7:3 (v/v)] investigated by using UHPLC-DAD-MSn and subsequent dereplication. The permeability of compounds present in PPX was evaluated using the Caco-2 monolayer permeation system. The influence of PPX on uropathogenic E. coli (UPEC) strain NU14 proliferation and against the bacterial adhesion to T24 epithelial cells was determined by turbidimetric assay and flow cytometry, respectively. The influence of the extract on the mitochondrial activity of T24 host cells was monitored by MTT assay. Results LC-MSn investigation and dereplication, indicated PPX extract to be dominated by a variety of flavonoids, with rutin as a major compound, and soyasaponin derivatives. Rutin, selected soyasaponins and fatty acids were shown to permeate the Caco-2 monolayer system, indicating potential bioavailability following oral intake. The extract did not influence the viability of T24 cells after 1.5h incubation at 2 mg/mL and UPEC. PPX significantly reduced the bacterial adhesion of UPEC to human bladder cells in a concentration-dependent manner (0.5–2 mg/mL). Detailed investigations by different incubation protocols indicated that PPX seems to interact with T24 cells, which subsequently leads to reduced recognition and adhesion of UPEC to the host cell membrane. Conclusions PPX is characterised by the presence of flavonoids (e.g. rutin) and saponins, from which selected compounds might be bioavailable after oral application, as indicated by the Caco-2 permeation experiments. Rutin and some saponins can be considered as potentially bioavailable after the oral intake. The concentration-dependent inhibition of bacterial adhesion of UPEC to T24 cells justifies the traditional use of Phaseoli pericarpium in the prevention and treatment of urinary tract infections.

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    Journal of Ethnopharmacology
    Article . 2021 . Peer-reviewed
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    Journal of Ethnopharmacology
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    https://dx.doi.org/10.17169/re...
    Other literature type . 2021
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      Journal of Ethnopharmacology
      Article . 2021 . Peer-reviewed
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    Authors: Jens Wiltfang; J. Porzig; Johannes Kornhuber; Paul Dieter Niedmann; +7 Authors

    Ethanol exerts its behavioural effects largely by interacting with receptors for brain neurotransmitters. However, the molecular mechanisms involving these interactions and the pathogenesis of alcohol-withdrawal symptomatology are still not well understood. Until recently, no data were available about homocysteine (Hcy) levels in acute alcohol intoxication of chronic alcoholics and in patients undergoing withdrawal from alcohol. Hcy, blood-alcohol concentrations, vitamins B6, B12, and folate concentrations were assessed in 29 chronic alcoholics, who underwent withdrawal from alcohol. We observed increased Hcy levels in most patients. Hcy levels steadily decreased during the observation period. We postulate that hyperhomocysteinaemia and excitatory amino acid neurotransmitters, by their agonism at the N-methyl-D-aspartate receptor, may partly mediate alcohol-associated withdrawal symptomatology. The importance of assessing serum Hcy levels in order to detect methylation deficiency in patients with chronic alcoholism and for possible therapeutic strategies is discussed.

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    Alcohol and Alcoholism
    Article . 2000 . Peer-reviewed
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      Alcohol and Alcoholism
      Article . 2000 . Peer-reviewed
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    Authors: David Tomé-Lourido; Dolores Ponte; María José Sampedro; Constantino Arce;

    The COVID-19 pandemic caused a great emotional impact on the general population, with specific consequences in the field of sport and physical activity. The present study was conducted to meet two objectives: (1) to investigate if the pandemic had an impact on competitive anxiety and if the sex and competitive level played a mediating role, and (2) to investigate if the relationship between emotional self-control and anxiety was affected by the pandemic. An amount of 608 Spanish athletes participated, divided into two cohorts, before and after the pandemic. Measurements of competitive anxiety and emotional self-control were made at both time points, which were processed by combining analysis of variance and regression models. Differences were found in competitive anxiety before and after COVID-19, with athlete’s competitive level acting as a mediating variable. For high-level athletes, there was an increase in anxiety, while for lower-level athletes, there was a decrease in anxiety. The pandemic seems to have led to higher levels of anxiety in elite athletes, compared to amateur athletes. There was no interaction between the sex of the participants and the cohort. Emotional self-control remained a significant predictor of low competitive anxiety levels.

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    Sustainability
    Article . 2023 . Peer-reviewed
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    Sustainability
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      Sustainability
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      Sustainability
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    Authors: Jun Jie Ong; Simon Gaisford; Edmont Stoyanov; Abdul Basit; +5 Authors

    In the past decade, prescriptions for opioid medicines have been exponentially increasing, instigating opioid abuse as a global health crisis associated with high morbidity and mortality. In particular, diversion from the intended mode of opioid administration, such as injecting and snorting the opioid, is a major problem that contributes to this epidemic. In light of this, novel formulation strategies are needed to support efforts in reducing the prevalence and risks of opioid abuse. Here, modified release tramadol printlets (3D printed tablets) with alcohol-resistant and abuse-deterrent properties were prepared by direct powder extrusion three-dimensional (3D) printing. The printlets were fabricated using two grades of hydroxypropylcellulose (HPC). Both formulations displayed strong ethanol-resistance and had moderate abuse-deterrent properties. Polyethylene oxide (PEO) was subsequently added into the formulations, which improved the printlets' resistance to physical tampering in nasal inhalation tests and delayed their dissolution in solvent extraction tests. Overall, this article reports for the first time the use of direct powder extrusion 3D printing to prepare drug products with both alcohol-resistant and abuse-deterrent properties. These results offer a novel approach for the safe and effective use of opioids that can contribute to the advantages that 3D printing provides in terms of on-demand dose personalisation.

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    International Journal of Pharmaceutics
    Article . 2020 . Peer-reviewed
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      International Journal of Pharmaceutics
      Article . 2020 . Peer-reviewed
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  • image/svg+xml art designer at PLoS, modified by Wikipedia users Nina, Beao, JakobVoss, and AnonMoos Open Access logo, converted into svg, designed by PLoS. This version with transparent background. http://commons.wikimedia.org/wiki/File:Open_Access_logo_PLoS_white.svg art designer at PLoS, modified by Wikipedia users Nina, Beao, JakobVoss, and AnonMoos http://www.plos.org/
    Authors: Fouad M.F. Elshaghabee; Fouad M.F. Elshaghabee; Wilhelm eBockelmann; Diana eMeske; +4 Authors

    Pour obtenir un aperçu spécifique des rôles que les micro-organismes pourraient jouer dans la stéatose hépatique non alcoolique (NAFLD), certaines bactéries intestinales et lactiques et une levure (Anaerostipes caccae, Bacteroides thetaiotaomicron, Bifidobacterium longum, Enterococcus fecalis, Escherichia coli, Lactobacillus acidophilus, Lactobacillus fermentum, Lactobacillus plantarum, Weissella confusa, Saccharomyces cerevisiae) ont été caractérisées par une chromatographie liquide haute performance pour la production d'éthanol lorsqu'elles sont cultivées sur différents glucides : hexoses (glucose et fructose), pentoses (arabinose et ribose), disaccharides (lactose et lactulose) et inuline. Les quantités les plus élevées d'éthanol ont été produites par S. cerevisiae, L. fermentum et W. confusa sur le glucose et par S. cerevisiae et W. confusa sur le fructose. En raison de la mannitol-déshydrogénase exprimée dans L. fermentum, la production d'éthanol sur le fructose a été significativement réduite (P < 0,05). Le pyruvate et le citrate, deux accepteurs d'électrons potentiels pour la régénération du NAD+/NADP+, ont considérablement réduit la production d'éthanol avec de l'acétate produit à la place dans L. fermentum cultivé sur glucose et W. confusa cultivé sur glucose et fructose, respectivement. Dans les boues fécales préparées à partir des matières fécales de quatre volontaires en surpoids, on a constaté que l'éthanol était produit lors de l'ajout de fructose. L'ajout d'A. caccae, L. acidophilus, L. fermentum, ainsi que de citrate et de pyruvate, respectivement, a aboli la production d'éthanol. Cependant, l'ajout de W. confusa a entraîné une augmentation significative (P < 0,05) de la production d'éthanol. Ces résultats indiquent que des micro-organismes comme W. confusa, une bactérie lactique hétéro-fermentaire, négative à la mannitol-déshydrogénase, peuvent favoriser la NAFLD par l'éthanol produit à partir de la fermentation du sucre, tandis que d'autres bactéries intestinales et des bactéries lactiques homo- et hétéro-fermentaires mais positives à la mannitol-déshydrogénase peuvent ne pas favoriser la NAFLD. En outre, nos études indiquent que les facteurs alimentaires interférant avec le microbiote gastro-intestinal et le métabolisme microbien peuvent être importants dans la prévention ou la promotion de la NAFLD. Para obtener información específica sobre los roles que podrían desempeñar los microorganismos en la enfermedad del hígado graso no alcohólico (NAFLD, por sus siglas en inglés), algunas bacterias intestinales y del ácido láctico y una levadura (Anaerostipes caccae, Bacteroides thetaiotaomicron, Bifidobacterium longum, Enterococcus fecalis, Escherichia coli, Lactobacillus acidophilus, Lactobacillus fermentum, Lactobacillus plantarum, Weissella confusa, Saccharomyces cerevisiae) se caracterizaron por cromatografía líquida de alto rendimiento para la producción de etanol cuando se cultivaron en diferentes carbohidratos: hexosas (glucosa y fructosa), pentosas (arabinosa y ribosa), disacáridos (lactosa y lactulosa) e inulina. Las cantidades más altas de etanol fueron producidas por S. cerevisiae, L. fermentum y W. confusa en glucosa y por S. cerevisiae y W. confusa en fructosa. Debido a la manitol-deshidrogenasa expresada en L. fermentum, la producción de etanol en fructosa se redujo significativamente (P < 0.05). El piruvato y el citrato, dos aceptores de electrones potenciales para la regeneración de NAD+/NADP+, redujeron drásticamente la producción de etanol con acetato producido en su lugar en L. fermentum cultivado en glucosa y W. confusa cultivado en glucosa y fructosa, respectivamente. En suspensiones fecales preparadas a partir de heces de cuatro voluntarios con sobrepeso, se encontró que el etanol se producía tras la adición de fructosa. La adición de A. caccae, L. acidophilus, L. fermentum, así como citrato y piruvato, respectivamente, abolió la producción de etanol. Sin embargo, la adición de W. confusa resultó en un aumento significativo (P < 0.05) de la producción de etanol. Estos resultados indican que microorganismos como W. confusa, una bacteria de ácido láctico hetero-fermentativa, negativa para manitol-deshidrogenasa, pueden promover NAFLD a través del etanol producido a partir de la fermentación de azúcar, mientras que otras bacterias intestinales y bacterias de ácido láctico homo- y hetero-fermentativas pero positivas para manitol-deshidrogenasa pueden no promover NAFLD. Además, nuestros estudios indican que los factores dietéticos que interfieren con la microbiota gastrointestinal y el metabolismo microbiano pueden ser importantes para prevenir o promover la EHGNA. To gain some specific insight into the roles microorganisms might play in non-alcoholic fatty liver disease (NAFLD), some intestinal and lactic acid bacteria and one yeast (Anaerostipes caccae, Bacteroides thetaiotaomicron, Bifidobacterium longum, Enterococcus fecalis, Escherichia coli, Lactobacillus acidophilus, Lactobacillus fermentum, Lactobacillus plantarum, Weissella confusa, Saccharomyces cerevisiae) were characterized by high performance liquid chromatography for production of ethanol when grown on different carbohydrates: hexoses (glucose and fructose), pentoses (arabinose and ribose), disaccharides (lactose and lactulose), and inulin. Highest amounts of ethanol were produced by S. cerevisiae, L. fermentum and W. confusa on glucose and by S. cerevisiae and W. confusa on fructose. Due to mannitol-dehydrogenase expressed in L. fermentum, ethanol production on fructose was significantly (P < 0.05) reduced. Pyruvate and citrate, two potential electron acceptors for regeneration of NAD+/NADP+, drastically reduced ethanol production with acetate produced instead in L. fermentum grown on glucose and W. confusa grown on glucose and fructose, respectively. In fecal slurries prepared from feces of four overweight volunteers, ethanol was found to be produced upon addition of fructose. Addition of A. caccae, L. acidophilus, L. fermentum, as well as citrate and pyruvate, respectively, abolished ethanol production. However, addition of W. confusa resulted in significantly (P < 0.05) increased production of ethanol. These results indicate that microorganisms like W. confusa, a hetero-fermentative, mannitol-dehydrogenase negative lactic acid bacterium, may promote NAFLD through ethanol produced from sugar fermentation, while other intestinal bacteria and homo- and hetero-fermentative but mannitol-dehydrogenase positive lactic acid bacteria may not promote NAFLD. Also, our studies indicate that dietary factors interfering with gastrointestinal microbiota and microbial metabolism may be important in preventing or promoting NAFLD. لاكتساب بعض الأفكار المحددة حول الأدوار التي قد تلعبها الكائنات الحية الدقيقة في مرض الكبد الدهني غير الكحولي (NAFLD)، تميزت بعض بكتيريا حمض الأمعاء واللاكتيك وخميرة واحدة (Anaerostipes caccae، Bacteroides thetaiotaomicron، Bifidobacterium longum، Enterococcus fecalis، Escherichia coli، Lactobacillus acidophilus، Lactobacillus fermentum، Lactobacillus plantarum، Weissella confusa، Saccharomyces cerevisiae) بتصوير سائل عالي الأداء لإنتاج الإيثانول عند زراعته على كربوهيدرات مختلفة: hexoses (الجلوكوز والفركتوز)، pentoses (الأرابينوز والريبوز)، disaccharides (اللاكتوز واللاكتولوز)، و inulin. تم إنتاج أعلى كميات من الإيثانول بواسطة S. cerevisiae و L. fermentum و W. confusa على الجلوكوز و S. cerevisiae و W. confusa على الفركتوز. بسبب نازعة هيدروجين المانيتول المعبر عنها في L. fermentum، انخفض إنتاج الإيثانول على الفركتوز بشكل كبير (P < 0.05). قلل البيروفات والسيترات، وهما مستقبلان محتملان للإلكترون لتجديد NAD +/NADP+، بشكل كبير من إنتاج الإيثانول مع الأسيتات المنتجة بدلاً من ذلك في L. fermentum المزروع على الجلوكوز و W. confusa المزروع على الجلوكوز والفركتوز، على التوالي. في الملاط البرازي الذي تم تحضيره من براز أربعة متطوعين يعانون من زيادة الوزن، وجد أن الإيثانول يتم إنتاجه عند إضافة الفركتوز. إضافة A. caccae، L. acidophilus، L. fermentum، وكذلك السترات والبيروفات، على التوالي، ألغت إنتاج الإيثانول. ومع ذلك، أدت إضافة W. confusa إلى زيادة كبيرة في إنتاج الإيثانول (P < 0.05). تشير هذه النتائج إلى أن الكائنات الحية الدقيقة مثل W. confusa، وهي بكتيريا حمض اللاكتيك السلبية غير المتجانسة، قد تعزز NAFLD من خلال الإيثانول المنتج من تخمير السكر، في حين أن البكتيريا المعوية الأخرى وبكتيريا حمض اللاكتيك الإيجابية غير المتجانسة ولكن غير المتجانسة قد لا تعزز NAFLD. أيضًا، تشير دراساتنا إلى أن العوامل الغذائية التي تتداخل مع الكائنات الحية الدقيقة في الجهاز الهضمي والتمثيل الغذائي الميكروبي قد تكون مهمة في منع أو تعزيز NAFLD.

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    Frontiers in Microbiology
    Article . 2016 . Peer-reviewed
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    Authors: David Caldevilla-Domínguez; Almudena Barrientos-Báez; Graciela Padilla-Castillo;

    The social confinement resulting from the COVID-19 crisis temporarily reduced greenhouse gas emissions. Although experts contend that the decrease in pollution rates was not drastic, some surveys detect growth in social concern about the climate. In this new climate-conscious environment, municipalities and local governments are promoting a new way of living and caring for cities, even before they can regain national and international freedom of movement. This work analyzes the connections between new climate awareness arising from the COVID-19 crisis, proposals of sustainable citizenship around the world, and its communication on Twitter to educate the new eco-conscious audience. The methodology mixes quantitative and qualitative analysis, using the Twitonomy Premium tool and the Twitter research tool with data extracted at the end of December 2020. Among the top ten most influential and active accounts, the results show educational institutions, local institutions, companies, neighborhoods, associations, and influencers. The impossibility of living in the city has not prevented citizen education and commitment to make real change for when that city and its citizens return to normality. However, this new normality must be different: more ecological, more responsible, more sustainable, and practiced from early childhood.

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    Sustainability
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    Authors: Jorge Montesinos; Anabel Gil; Consuelo Guerri;

    BackgroundWe previously showed that, by activating innate immune receptors Toll‐like 4 (TLR4), adolescent intermittent ethanol (EtOH) exposure causes neuroinflammation, myelin damage, and behavioral dysfunctions. Recent findings reveal that clinically used opioid antagonists naltrexone (NT) and naloxone (NX) inhibit opioid‐induced TLR4 signaling and that NT, NX, and nalmefene (NF), the 6‐methylene derivative of NX, are able to reduce alcohol drinking escalation.MethodsNF (0.1 mg/kg, intraperitoneally) was injected 1 hour prior to EtOH (3 g/kg, intraperitoneally) following intermittent treatment in female (PND35) adolescent mice. Inflammatory molecules, myelin proteins, and apoptotic markers were assessed in the prefrontal cortex (PFC) and striatum/nucleus accumbens (STR/NAcc). The effect of NF on alcohol drinking preference was evaluated in both the wild‐type and TLR4 knockout (KO) adolescent mice. Using astroglial cells, the inhibitory potential of NT, NX, and NF on lipopolysaccharide (LPS), or the EtOH‐triggered TLR4 response, was compared.ResultsOur findings indicate that NF prevents the up‐regulation of cytokines (IL‐1β, IL‐17A, TNF‐α), chemokines (MCP‐1, MIP‐1, KC), and pro‐inflammatory mediators (iNOS, COX‐2), along with myelin damage and apoptotic events, in both PFC and STR/NAcc. NF also abolishes EtOH‐induced escalation of alcohol preference/consumption, but has no effect when administered to TLR4‐KO mice. In vitro experiments indicate that NX and NF inhibit TLR4 activation upon LPS or EtOH stimulation. Immunofluorescence studies and lipid rafts isolation show that NF is able to prevent TLR4 translocation to lipid rafts/caveolae in astrocytes.ConclusionsThese results suggest that NF prevents neuroinflammation and brain damage by blocking the TLR4 response and also support the role of central pro‐inflammatory immune signaling in the modulation of alcohol consumption/addiction.

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    Alcoholism Clinical and Experimental Research
    Article . 2017 . Peer-reviewed
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      Alcoholism Clinical and Experimental Research
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    Authors: Marisa Silva; Vijaya Pratheepa; Luis Botana; Vitor Vasconcelos;

    Harmful Algal Blooms (HAB) are complex to manage due to their intermittent nature and their severe impact on the economy and human health. The conditions which promote HAB have not yet been fully explained, though climate change and anthropogenic intervention are pointed as significant factors. The rise of water temperature, the opening of new sea canals and the introduction of ship ballast waters all contribute to the dispersion and establishment of toxin-producing invasive species that promote the settling of emergent toxins in the food-chain. Tetrodotoxin, ciguatoxin, palytoxin and cyclic imines are commonly reported in warm waters but have also caused poisoning incidents in temperate zones. There is evidence that monitoring for these toxins exclusively in bivalves is simplistic and underestimates the risk to public health, since new vectors have been reported for these toxins and as well for regulated toxins such as PSTs and DSTs. In order to avoid public health impacts, there is a need for adequate monitoring programs, a need for establishing appropriate legislation, and a need for optimizing effective methods of analysis. In this review, we will compile evidence concerning emergent marine toxins and provide data that may indicate the need to restructure the current monitoring programs of HAB.

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    Toxins
    Article . 2015 . Peer-reviewed
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    Article . 2016
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    Authors: Román, Juan; Colell Riera, Anna; Blasco, Carmen; Caballería, Juan; +3 Authors

    The oxidative metabolism of ethanol by the cytochrome P450 2E1 (CYP2E1) has been recognized to contribute to the ethanol-induced deleterious effects through the induction of oxidative stress. This study compared the effect of ethanol and acetaldehyde in the induction of oxidative stress and activation of transcription factors nuclear factor-κB (NF-κB) and activating protein 1 (AP-1) in HepG2 cells, which do not express CYP2E1, and HepG2 cells transfected with CYP2E1 (E47 cells). Neither ethanol (80 mmol/L) nor acetaldehyde (25-200 μmol/L) caused oxidative stress in HepG2 cells, an effect that was independent of blocking reduced glutathione (GSH) synthesis with buthionine-l -sulfoximine (BSO). However, BSO preincubation caused an overproduction of peroxides and activation of NF-κB and AP-1 in E47 cells even in the absence of ethanol. Furthermore, the incubation of E47 cells with ethanol (80 mmol/L for up to 5 days) depleted cellular GSH stores in both cytosol and mitochondria, reflecting the induction of oxidative stress. Ethanol activated NF-κB and AP-1 in E47 cells, an effect that was prevented by 4-methylpyrazole, potentiated by cyanamide, and attenuated by trolox C. Interestingly, however, despite the inability of acetaldehyde to induce oxidative stress in HepG2, acetaldehyde activated NF-κB and AP-1; in contrast, ethanol failed to activate these transcription factors in HepG2. Thus, our findings indicate that activation of NF-κB and AP-1 by ethanol and acetaldehyde occurs through distinct mechanisms. CYP2E1 is indispensable in the induction of oxidative stress from ethanol, whereas the activation of NF-κB and AP-1 by acetaldehyde is independent of oxidative stress.

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    Hepatology
    Article . 1999 . Peer-reviewed
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    Article . 1999
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    Article . 1999 . Peer-reviewed
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      Hepatology
      Article . 1999 . Peer-reviewed
      License: Wiley TDM
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      Hepatology
      Article . 1999
      Hepatology
      Article . 1999 . Peer-reviewed
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    Authors: Birte Sievers; Jörg Hausdorf; Volkmar Jansson; Susanne Mayer-Wagner; +5 Authors

    AbstractThe aim of this study was to determine if extracorporeal shock wave therapy (ESWT) in vivo affects the structural integrity of articular cartilage. A single bout of ESWT (1500 shock waves of 0.5 mJ/mm2) was applied to femoral heads of 18 adult Sprague–Dawley rats. Two sham‐treated animals served as controls. Cartilage of each femoral head was harvested at 1, 4, or 10 weeks after ESWT (n = 6 per treatment group) and scored on safranin‐O‐stained sections. Expression of tenascin‐C and chitinase 3‐like protein 1 (Chi3L1) was analyzed by immunohistochemistry. Quantitative real‐time polymerase chain reaction (PCR) was used to examine collagen (II)α1 (COL2A1) expression and chondrocyte morphology was investigated by transmission electron microscopy no changes in Mankin scores were observed after ESWT. Positive immunostaining for tenascin‐C and Chi3L1 was found up to 10 weeks after ESWT in experimental but not in control cartilage. COL2A1 mRNA was increased in samples 1 and 4 weeks after ESWT. Alterations found on the ultrastructural level showed expansion of the rough‐surfaced endoplasmatic reticulum, detachment of the cell membrane and necrotic chondrocytes. Extracorporeal shock waves caused alterations of hyaline cartilage on a molecular and ultrastructural level that were distinctly different from control. Similar changes were described before in the very early phase of osteoarthritis (OA). High‐energy ESWT might therefore cause degenerative changes in hyaline cartilage as they are found in initial OA. © 2010 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 28:1050–1056, 2010

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    Journal of Orthopaedic Research®
    Article . 2010 . Peer-reviewed
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      Journal of Orthopaedic Research®
      Article . 2010 . Peer-reviewed
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    Authors: Dominik Popowski; Karolina A. Pawłowska; Melanie Deipenbrock; Andreas Hensel; +4 Authors

    Ethnopharmacological relevance Phaseaoli pericarpium (bean pods) is a pharmacopeial plant material traditionally used as a diuretic and antidiabetic agents. Diuretic activity of pod extracts was reported first in 1608. Since then Phaseoli pericarpium tea figures in many textbooks as medicinal plant material used by patients. Aim of the study Despite the traditional use of extracts from Phaseolium vulgaris pericarp, limited information is available on bioactivity, chemical composition, and bioavailability of such preparations. The following study aimed to investigate the phytochemical composition, the in vitro permeability of selected extract's constituents over the Caco-2 permeation system, and potential antivirulence activity against uropathogenic Escherichia coli of a hydroalcoholic Phaseoli pericarpium extract (PPX) in vitro to support its traditional use as a remedy used in urinary tract infections. Material and methods The chemical composition of the extract PPX [ethanol:water 7:3 (v/v)] investigated by using UHPLC-DAD-MSn and subsequent dereplication. The permeability of compounds present in PPX was evaluated using the Caco-2 monolayer permeation system. The influence of PPX on uropathogenic E. coli (UPEC) strain NU14 proliferation and against the bacterial adhesion to T24 epithelial cells was determined by turbidimetric assay and flow cytometry, respectively. The influence of the extract on the mitochondrial activity of T24 host cells was monitored by MTT assay. Results LC-MSn investigation and dereplication, indicated PPX extract to be dominated by a variety of flavonoids, with rutin as a major compound, and soyasaponin derivatives. Rutin, selected soyasaponins and fatty acids were shown to permeate the Caco-2 monolayer system, indicating potential bioavailability following oral intake. The extract did not influence the viability of T24 cells after 1.5h incubation at 2 mg/mL and UPEC. PPX significantly reduced the bacterial adhesion of UPEC to human bladder cells in a concentration-dependent manner (0.5–2 mg/mL). Detailed investigations by different incubation protocols indicated that PPX seems to interact with T24 cells, which subsequently leads to reduced recognition and adhesion of UPEC to the host cell membrane. Conclusions PPX is characterised by the presence of flavonoids (e.g. rutin) and saponins, from which selected compounds might be bioavailable after oral application, as indicated by the Caco-2 permeation experiments. Rutin and some saponins can be considered as potentially bioavailable after the oral intake. The concentration-dependent inhibition of bacterial adhesion of UPEC to T24 cells justifies the traditional use of Phaseoli pericarpium in the prevention and treatment of urinary tract infections.

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    Journal of Ethnopharmacology
    Article . 2021 . Peer-reviewed
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    Journal of Ethnopharmacology
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    https://dx.doi.org/10.17169/re...
    Other literature type . 2021
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      Journal of Ethnopharmacology
      Article . 2021 . Peer-reviewed
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      Journal of Ethnopharmacology
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      Other literature type . 2021
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    Authors: Jens Wiltfang; J. Porzig; Johannes Kornhuber; Paul Dieter Niedmann; +7 Authors

    Ethanol exerts its behavioural effects largely by interacting with receptors for brain neurotransmitters. However, the molecular mechanisms involving these interactions and the pathogenesis of alcohol-withdrawal symptomatology are still not well understood. Until recently, no data were available about homocysteine (Hcy) levels in acute alcohol intoxication of chronic alcoholics and in patients undergoing withdrawal from alcohol. Hcy, blood-alcohol concentrations, vitamins B6, B12, and folate concentrations were assessed in 29 chronic alcoholics, who underwent withdrawal from alcohol. We observed increased Hcy levels in most patients. Hcy levels steadily decreased during the observation period. We postulate that hyperhomocysteinaemia and excitatory amino acid neurotransmitters, by their agonism at the N-methyl-D-aspartate receptor, may partly mediate alcohol-associated withdrawal symptomatology. The importance of assessing serum Hcy levels in order to detect methylation deficiency in patients with chronic alcoholism and for possible therapeutic strategies is discussed.

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    Alcohol and Alcoholism
    Article . 2000 . Peer-reviewed
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      Alcohol and Alcoholism
      Article . 2000 . Peer-reviewed
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    Authors: David Tomé-Lourido; Dolores Ponte; María José Sampedro; Constantino Arce;

    The COVID-19 pandemic caused a great emotional impact on the general population, with specific consequences in the field of sport and physical activity. The present study was conducted to meet two objectives: (1) to investigate if the pandemic had an impact on competitive anxiety and if the sex and competitive level played a mediating role, and (2) to investigate if the relationship between emotional self-control and anxiety was affected by the pandemic. An amount of 608 Spanish athletes participated, divided into two cohorts, before and after the pandemic. Measurements of competitive anxiety and emotional self-control were made at both time points, which were processed by combining analysis of variance and regression models. Differences were found in competitive anxiety before and after COVID-19, with athlete’s competitive level acting as a mediating variable. For high-level athletes, there was an increase in anxiety, while for lower-level athletes, there was a decrease in anxiety. The pandemic seems to have led to higher levels of anxiety in elite athletes, compared to amateur athletes. There was no interaction between the sex of the participants and the cohort. Emotional self-control remained a significant predictor of low competitive anxiety levels.

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    Sustainability
    Article . 2023 . Peer-reviewed
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    Article . 2023
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      Sustainability
      Article . 2023 . Peer-reviewed
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    Authors: Jun Jie Ong; Simon Gaisford; Edmont Stoyanov; Abdul Basit; +5 Authors

    In the past decade, prescriptions for opioid medicines have been exponentially increasing, instigating opioid abuse as a global health crisis associated with high morbidity and mortality. In particular, diversion from the intended mode of opioid administration, such as injecting and snorting the opioid, is a major problem that contributes to this epidemic. In light of this, novel formulation strategies are needed to support efforts in reducing the prevalence and risks of opioid abuse. Here, modified release tramadol printlets (3D printed tablets) with alcohol-resistant and abuse-deterrent properties were prepared by direct powder extrusion three-dimensional (3D) printing. The printlets were fabricated using two grades of hydroxypropylcellulose (HPC). Both formulations displayed strong ethanol-resistance and had moderate abuse-deterrent properties. Polyethylene oxide (PEO) was subsequently added into the formulations, which improved the printlets' resistance to physical tampering in nasal inhalation tests and delayed their dissolution in solvent extraction tests. Overall, this article reports for the first time the use of direct powder extrusion 3D printing to prepare drug products with both alcohol-resistant and abuse-deterrent properties. These results offer a novel approach for the safe and effective use of opioids that can contribute to the advantages that 3D printing provides in terms of on-demand dose personalisation.

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    International Journal of Pharmaceutics
    Article . 2020 . Peer-reviewed
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      International Journal of Pharmaceutics
      Article . 2020 . Peer-reviewed
      License: Elsevier TDM
      Data sources: Crossref
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