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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.

AbstractThe co‐occurrence of chronic pain and alcohol use disorders (AUDs) involves complex interactions between genetic and neurophysiological aspects, and the research has reported mixed findings when they both co‐occur. There is also an indication of a gender‐dependent effect; males are more likely to use alcohol to cope with chronic pain problems than females. Recently, a new conceptualization has emerged, proposing that the negative affective component of pain drives and maintains alcohol‐related behaviors. We studied in a longitudinal fashion alterations in alcohol drinking patterns and pain thresholds in a mouse model of chronic neuropathic pain in a sex‐dependent manner. Following partial denervation (spared nerve injury [SNI]), stimulus‐evoked pain responses were measured before chronic alcohol consumption, during drinking, during a deprivation phase, and following an episode of excessive drinking. During the course of alcohol drinking, we observed pronounced sex differences in pain thresholds. Male mice showed a strong increase in pain thresholds, suggesting an analgesic effect induced by alcohol over time, an effect that was not observed in female mice. SNI mice did not differ from sham‐operated controls in baseline alcohol consumption. However, following a deprivation phase and the reintroduction of ethanol, male SNI mice but not female mice showed more pronounced excessive drinking than controls. Finally, we observed decreased central ethanol sensitivity in male SNI mice but not in females. Together with our finding, that ethanol is able to decrease a pain‐induced negative affective memory we come to following conclusion. We propose that a lower sensitivity to the intoxicating effects of alcohol together with the ability of alcohol to reduce the negative affective component of pain may explain the higher co‐occurrence of AUD in male chronic pain patients.

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

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.

It has been estimated that more than 80% of alcoholics are also nicotine dependent and that, vice versa, the rate of alcoholism is substantially increased by a factor of 4-10 in the nicotine-dependent population. However, the cause for this very high degree of comorbidity is still largely unknown. At the molecular and cellular level, both drugs have very different mechanisms of action. Nicotine specifically activates ligand-gated ion channels in the brain, which are normally gated by acetylcholine, while alcohol interacts with various neurotransmitter receptors. Despite this diversity, both drugs seem to engage the endogenous opioid system as a modulator of some of its pharmacological effect. An acute exposure to nicotine or alcohol leads to a release of opioid peptides in specific brain regions, thus resulting in an activation of their corresponding receptors. If the brain is exposed repeatedly or chronically to these drugs, adaptive changes in the level and expression of opioid peptides and receptors occur. These adaptive changes are thought to contribute to the homeostatic or allostatic adaptations of the brain, which have been associated with drug dependence. This review summarizes pharmacological and genetic studies in animal models and in humans that have addressed the role of specific opioid peptides and receptors in various stages of the addiction process.

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.

Miniaturization of electrochemical detection ofPseudomonas aeruginosaquorum sensing molecules at a liquid/liquid interface through facilitated proton transfer reactions.