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In view of the importance of mesangial extracellular matrix (ECM) accumulation in the pathogenesis of diabetic glomerulosclerosis, we investigated 1) the effects of high glucose on ECM production by rat glomerular mesangial cells in culture (study A) and 2) the mechanisms underlying these effects, particularly the role of high sugar levels irrespective of intracellular metabolism (study B1) and of excess glucose disposal via the polyol pathway and associated biochemical alterations (study B2). Cells were cultured for 4 weeks, through six to eight passages, under the experimental conditions indicated below and, at each passage, the levels of fibronectin (FN), laminin (LAM), and collagen types I (C-I), III (C-III), IV (C-IV), and VI (C-VI) in media and cell extracts were quantified by an enzyme immunoassay. In study A, medium and cell content of matrix were assessed, together with [3H]leucine and [3H]thymidine incorporation into monolayers, polyol, fructose, and myo-inositol levels and the cytosolic redox state, in cells grown in high (30 mM) D-glucose or iso-osmolar mannitol versus cells cultured in normal (5.5 mM) D-glucose. FN, LAM, C-IV, and C-VI accumulation, but not C-I and C-III accumulation, was increased by 30 mM glucose, but not by iso-osmolar mannitol, when compared with 5.5 mM glucose, starting at week 2 and, except for C-VI, persisting throughout the remaining 2 weeks, whereas no change was observed in the measured indexes of total protein synthesis and DNA synthesis/cell proliferation. At any time point, polyol levels were increased, whereas myo-inositol was reduced by high glucose; in cells grown under elevated glucose concentrations, the lactate/pyruvate (L/P) ratio, an index of the cytosolic redox state, progressively increased. In study B1, the effects of high D-glucose were compared with those of iso-osmolar concentrations of sugars that are partly or not metabolized but are capable of inducing nonenzymatic glycosylation, such as D-galactose and L-glucose, and of mannitol, which does not enter the cell. Both D-galactose and L-glucose, but not mannitol, partly mimicked D-glucose-induced ECM overproduction. Although D-galactose is metabolized via the polyol pathway and alters the cytosolic redox state, ECM changes induced by high galactose were not prevented by the use of an aldose reductase inhibitor (ARI), Alcon 1576 (14 microM).(ABSTRACT TRUNCATED AT 400 WORDS)

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The hot water (HW) extraction of Norway spruce (Picea abies [Karst.]) delivers condensed tannins and considerable amounts of other compounds. Yield and composition of the HW extracts were investigated as a function of natural weathering for up to 15 months. Total phenol monomers and oligomers were detected by Folin-Ciocaltau assay after fractionation by solid phase extraction (SPE). Procyanidins (PC) were determined by HPLC-UV after acid thiolysis and carbohydrates by HPLC combined with acid hydrolysis. Topochemistry of the bark before and after extraction was investigated by UV-microspectrometry (UMSP) and non-extractable PC analyzed by direct thiolysis on the bark. The influence of the parameters on the yield and composition of the extracts were evaluated, such as the extraction temperature, time and the addition of sodium sulfate and urea. Prolonged weathering resulted in a considerable decrease of the total extraction yield, partly because of leaching of phenolic monomers, mono- and oligosaccharides. The yield of phenolic oligomers also decreased at a moderate rate, while the yield of polysaccharides (pectins) was almost stable. Non-extractable and non-leachable compounds deposited in the cell lumens represent the majority of the phenolic extractives in spruce bark. Sequential extractions performed at increasing temperature proved to be a suitable method for the recovery of tannin-rich extracts.

(1)Tannins have shown antifungal effects and have been considered a potential natural compound for wood preservation. Extracts produced from softwood bark contain both tannins and non-tannin compounds, which may reduce the effectiveness of tannin used as a wood preservative. The purpose of this research is to study the environmental impact of hot water extraction, identify the hot spots within the tannin cradle-to-gate life cycle and give suggestions to optimize its environmental profile. Different extraction and postextraction scenarios of tannin production are compared using the life-cycle assessment method. Experiments were designed to study the tannin yield under different extraction scenarios; the post-extraction scenario analysis was based on literature review. The results show that the extract drying process is the primary contributor to the environmental impact of tannin production. Both preliminary cold water extraction and ultrafiltration after extraction are beneficial as they have fewer non-tannin compounds in the final products; however, preliminary cold water extraction had a considerably lower environmental performance. Successive extractions using fresh water at each cycle increased the total tannin yield, but increased the environmental burden. Using only evaporation to obtain a desired tannin concentration is not environmentally efficient. This paper provides a quantified environmental analysis for the development of tannin-treated wood products and discusses the different tannin extraction scenarios from an environmental point of view.