Structure and Function of Stony Coral Intraskeletal Polysaccharides

Polysaccharides represent a main weight fraction of the intraskeletal organic matrix of corals, but their structure, as well as their function in the calcification process, has been poorly investigated. This communication shows by a combination of techniques (nuclear magnetic resonance, Fourier transform infrared, and monosaccharide composition) that their key component is a 1\u21923 \u3b2-d glucuronic acid polymer and evidences its influence in vitro in the calcification process

Foam formation in lakes: a long term analysis conducted on Lake Maggiore (northern Italy)

The causes and origin of foams in lakes have been rarely object of studies, although this phenomenon may cause problems to touristic or fisheries activities and imply a possible risk to human health. The formation of foams in the aquatic environment is due to the accumulation of surfaceactive compounds (surfactants) at the air-water interface joined with the mechanic action of a forcing (such as wind or waves) able to inject gas bubbles in the upper water layer. Surfactants can be either of natural or synthetic origin. Fulvic or humic acids, lipidic, proteic or colloidal substances are examples of natural surfactants that can promote foam formation, while man-made foams are generally due to the release of phosphates from agriculture and/or to the presence of organic and inorganic detergents. A comprehensive analysis of three foam episodes in Lake Maggiore (2007, 2008 and 2010) has been undertaken to identify their origin, causes and to unravel most likely factors triggering foam formation. At this scope, a long term (2000-2013) analysis of phytoplankton biovolumes and meteorological and hydrological anomalies has been performed together with the chemical characterization of foams. Foam resulted of endogenous origin, related to phytoplankton biomass degradation. The long term analysis highlighted atypical warm temperatures and residual lake stratification in winter in two of the years of foam events, coupled with exceptional Bacillariophyceae blooms in spring. Tabellaria flocculosa mostly contributed in terms of biomass in 2007 and 2008, but not in 2010, and overall total algal biomass seemed a better predictor of the risk of foam formation. Foam events occurred from July to December, driven by atypically high windy conditions, and congruently with the time needed to degrade biomass into surfactant compounds. A co-occurrence of different factors resulted essential to generate foams, and climate changes likely contributed to enhance their occurrence in Lake Maggiore

Cytotoxic effect of Agaricus bisporus and Lactarius rufus β-d-glucans on HepG2 cells

AbstractThe cytotoxic activity of β-d-glucans isolated from Agaricus bisporus and Lactarius rufus fruiting bodies was evaluated on human hepatocellular carcinoma cells (HepG2). NMR and methylation analysis suggest that these β-d-glucans were composed of a linear (1→6)-linked and a branched (1→3), (1→6)-linked backbone, respectively. They both decreased cell viability at concentrations of up to 100μgmL−1, as shown by MTT assay. The amount of LDH released and the analysis of cell morphology corroborated these values and also showed that the β-d-glucan of L. rufus was more cytotoxic to HepG2 cells than that of A. bisporus. The treatment of HepG2 cells with L. rufus and A. bisporus β-d-glucans at a dose of 200μgmL−1 for 24h promoted an increase of cytochrome c release and a decrease of ATP content, suggesting that these polysaccharides could promote cell death by apoptosis. Both β-d-glucans were tested against murine primary hepatocytes at a dose of 200μgmL−1. The results suggest that the L. rufus β-d-glucan was as cytotoxic for hepatocytes as for HepG2 cells, whereas the A. bisporus β-d-glucan, under the same conditions, was cytotoxic only for HepG2 cells, suggesting cell selectivity. These results open new possibilities for use of mushroom β-d-glucans in cancer therapy

Rhamnogalacturonan from Ilex paraguariensis: A potential adjuvant in sepsis treatment

AbstractThe present study evaluated the anti-inflammatory activity of a polysaccharide from maté, using a clinically relevant model of sepsis induced by cecal ligation and puncture (CLP). A polysaccharide from maté (SPI) was obtained from aqueous extraction followed by fractionation, being identified as a rhamnogalacturonan with a main chain of →4)-6-OMe-α-d-GalpA-(1→ groups, interrupted by α-l-Rhap units, substituted by a type I arabinogalactan. SPI was tested against induced-polymicrobial sepsis, at doses of 3, 7 and 10mg/kg. Via oral administration, SPI prevented the late mortality of infected mice by a rate of 60% at 10mg/kg, in comparison with untreated mice Dexamethasone, used as positive control, was slightly less effective, with an overall survival rate of 16.7% of mice at the end of the observation period. SPI also affected neutrophil influx, avoiding its accumulation in lungs, and significantly decreased tissue expression of iNOS and COX-2. In this context, maté is a potential nutraceutical, and its polysaccharide a promising adjuvant for sepsis treatment, being consumed as tea-like beverages with no related adverse effects

Antinociception of β-d-glucan from Pleurotus pulmonarius is possibly related to protein kinase C inhibition

Abstractβ-d-Glucan, a polysaccharide isolated from an edible mushroom Pleurotus pulmonarius (Fr.) Quel., presented antinociceptive activity in mice. This study evaluated the involvement of transient receptor potential (TRP) channels and protein kinase C (PKC) on antinociceptive effect of a (1→3),(1→6)-linked β-d-glucan (GL) in mice. Intraperitoneal administration of GL potently inhibited nociceptive responses induced by intraplantar injections of capsaicin, cinnamaldehyde, menthol, acidified saline and phorbol myristate acetate (PMA). Moreover, Western blot analysis revealed that GL treatment also prevented PMA-induced PKCɛ activation. Collectively, present results demonstrate that GL could constitute an attractive molecule of interest for the development of new analgesic drugs

Pressurized Liquid (PLE) Truffle Extracts Have Inhibitory Activity on Key Enzymes Related to Type 2 Diabetes (α-Glucosidase and α-Amylase)

An optimized PLE method was applied to several truffle species using three different solvent mixtures to obtain bioactive enriched fractions. The pressurized water extracts contained mainly (1 ? 3),(1 ? 6)-?-D-glucans, chitins, and heteropolymers with galactose and mannose in their structures. The ethanol extracts included fatty acids and fungal sterols and others such as brassicasterol and stigmasterol, depending on the species. They also showed a different fatty acid lipid profile depending on the solvent utilized and species considered. Ethanol:water extracts showed interesting lipids and many phenolic compounds; however, no synergic extraction of compounds was noticed. Some of the truffle extracts were able to inhibit enzymes related to type 2 diabetes; pressurized water extracts mainly inhibited the ?-amylase enzyme, while ethanolic extracts were more able to inhibit ?-glucosidase. Tuber brumale var. moschatum and T. aestivum var. uncinatum extracts showed an IC50 of 29.22 mg/mL towards ?-amylase and 7.93 mg/mL towards ?-glucosidase. Thus, use of the PLE method allows o bioactive enriched fractions to be obtained from truffles with antidiabetic properties.This research was funded by the fellowship Ibercaja-CAI Estancias de Investigación number CA 1/20. The research leading to these results has received funding from the European Union under “Horizon 2020—the Framework Programme for Research and Innovation (2014–2020)”. Grant Agreement of the Project: “Innovation in truffle cultivation, preservation, processing and wild truffle resources management”, INTACT, Project number 101007623.trufflesβ-D-glucansergosterolfatty acidsantidiabeticamylaseglucosidasePublishe

A robust method to quantify low molecular weight contaminants in heparin: detection of tris(2-n-butoxyethyl) phosphate

Recently, oversulfated chondroitin sulfate (OSCS) was identified in contaminated heparin preparations, which were linked to several adverse clinical events and deaths. Orthogonal analytical techniques, namely nuclear magnetic resonance (NMR) and capillary electrophoresis (CE), have since been applied by several authors for the evaluation of heparin purity and safety. NMR identification and quantification of residual solvents and non-volatile low molecular contaminants with USP acceptance levels of toxicity was achieved 40-fold faster than the traditional GC-headspace technique, which takes similar to 120 min against similar to 3 min to obtain a (1)H NMR spectrum with a signal/noise ratio of at least 1000/1. the procedure allowed detection of Class 1 residual solvents at 2 ppm and quantification was possible above 10 ppm. 2D NMR techniques (edited-HSQC (1)H/(13)C) permitted visualization of otherwise masked EDTA signals at 3.68/59.7 ppm and 3.34/53.5 ppm, which may be overlapping mononuclear heparin signals, or those of ethanol and methanol. Detailed NMR and ESI-MS/MS studies revealed a hitherto unknown contaminant, tris(2-n-butoxyethyl) phosphate (TBEP), which has potential health risks.Brazilian agency Fundacao AraucariaBrazilian agency FINEP (PRONEX-CARBOIDRATOS, PADCT II/SBIO)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Univ Fed Parana, Dept Bioquim & Biol Mol, BR-81531980 Curitiba, PR, BrazilIst Ric Chim & Biochim G Ronzoni, I-20133 Milan, ItalyUniversidade Federal de São Paulo, Dept Bioquim & Biol Mol, BR-04044020 São Paulo, SP, BrazilUniv Liverpool, Sch Biol Sci, Liverpool L69 7ZB, Merseyside, EnglandUniversidade Federal de São Paulo, Dept Bioquim & Biol Mol, BR-04044020 São Paulo, SP, BrazilWeb of Scienc

Polysaccharides from Agaricus bisporus and Agaricus brasiliensis show similarities in their structures and their immunomodulatory effects on human monocytic THP-1 cells

<p>Abstract</p> <p>Background</p> <p>Mushroom polysaccharides have traditionally been used for the prevention and treatment of a multitude of disorders like infectious illnesses, cancers and various autoimmune diseases. Crude mushroom extracts have been tested without detailed chemical analyses of its polysaccharide content. For the present study we decided to chemically determine the carbohydrate composition of semi-purified extracts from 2 closely related and well known basidiomycete species, i.e. <it>Agaricus bisporus </it>and <it>A. brasiliensis </it>and to study their effects on the innate immune system, in particular on the <it>in vitro </it>induction of pro-inflammatory cytokines, using THP-1 cells.</p> <p>Methods</p> <p>Mushroom polysaccharide extracts were prepared by hot water extraction and precipitation with ethanol. Their composition was analyzed by GC-MS and NMR spectroscopy. PMA activated THP-1 cells were treated with the extracts under different conditions and the production of pro-inflammatory cytokines was evaluated by qPCR.</p> <p>Results</p> <p>Semi-purified polysaccharide extracts of <it>A. bisporus </it>and <it>A. brasiliensis </it>(= <it>blazei</it>) were found to contain (1→6),(1→4)-linked α-glucan, (1→6)-linked β-glucan, and mannogalactan. Their proportions were determined by integration of ¹H-NMR signs, and were considerably different for the two species. <it>A. brasiliensis </it>showed a higher content of β-glucan, while <it>A. bisporus </it>presented mannogalactan as its main polysaccharide. The extracts induced a comparable increase of transcription of the pro-inflammatory cytokine genes IL-1β and TNF-α as well as of COX-2 in PMA differentiated THP-1 cells. Pro-inflammatory effects of bacterial LPS in this assay could be reduced significantly by the simultaneous addition of <it>A. brasiliensis </it>extract.</p> <p>Conclusions</p> <p>The polysaccharide preparations from the closely related species <it>A. bisporus </it>and <it>A. brasiliensis </it>show major differences in composition: <it>A. bisporus </it>shows high mannogalactan content whereas <it>A. brasiliensis </it>has mostly β-glucan. Semi-purified polysaccharide extracts from both <it>Agaricus </it>species stimulated the production of pro-inflammatory cytokines and enzymes, while the polysaccharide extract of <it>A. brasiliensis </it>reduced synthesis of these cytokines induced by LPS, suggesting programmable immunomodulation.</p

Commercial establishment

A presente dissertação aborda os principais aspectos relacionados ao estabelecimento empresarial no ordenamento jurídico pátrio. O estudo fundamentou-se na doutrina nacional e, mormente na fecunda doutrina italiana, a qual teve um papel fundamental neste trabalho. No primeiro capítulo, analisaram-se os alicerces do direito comercial: o empresário, a empresa e o estabelecimento. Em seguida, estudou-se o estabelecimento empresarial, definindo-o, bem como sua natureza jurídica, elementos e outros aspectos. No terceiro capítulo, os princípios informadores do estabelecimento foram abordados. Em seguida, analisou-se a origem do instituto e a doutrina estrangeira, dando-se ênfase à doutrina italiana em virtude da semelhança entre os ordenamentos. Por derradeiro, o aspecto dinâmico do estabelecimento foi estudado. O estabelecimento empresarial foi abordado como objeto de negócios jurídicos translativos e constitutivos. Sempre à luz da doutrina italiana e da nacional.The present dissertation had the object to study the most important aspects related to the commercial establishment in the brazilian legal system. The study was based on national doctrinal research and mostly on productive Italian doctrine. The Italian legislation also had an import aspect in this work. In the first chapter, the foundations of commercial law were analyzed: enterpriser, enterprise and establishment. After that, commercial establishment was defined, and, as well, as its legal nature, elements and other aspects. In the third chapter, the principles that inform the commercial establishment were study. Finally, the dynamic aspect of commercial establishment was analyzed. The commercial establishment was studied as object in legal transactions. Always in regard to Italian and national doctrine

Methylation-GC-MS analysis of arabinofuranose- and galactofuranose-containing structures: rapid synthesis of partially O-methylated alditol acetate standards

Arabinose and galactose were treated with MeOH containing traces of H2SO4 or HCl at 25ºC to give mixtures of their methyl alpha- and beta-furanosides, as shown by 1D and 2D nuclear magnetic resonance (NMR). Oxidation of the Me alpha,beta-Araf mixture with NaIO4 preferentially oxidised the beta-isomer, to give pure Me alpha-Araf . Each product was progressively O-methylated using the Purdie reagent (MeI/Ag2O) at 25ºC and resulting mixtures of partially methylated glycosides (PMGs) were rapidly assayed by thin layer chromatography (TLC) first to favour higher yields of mono-O-methyl derivatives and later for products with higher degrees of methylation. The products were converted to complex mixtures of partially O-methylated alditol acetate derivatives (PMAAs) by successive hydrolysis, reduction with NaBD4, and acetylation. These can be used as gas chromatography-mass spectrometry (GC-MS) standards in methylation analysis of complex carbohydrates containing arabinofuranosyl and galactofuranosyl units. Of particular interest were the retention times and electron impact MS of the difficult to prepare alditol acetates of 5,6-Me2Gal, 2,5-Me2Gal, 2,5,6-Me3Gal, 3,5,6-Me3Gal, 5-MeAra, 2,5-Me2Ara, and 3,5-Me2Ara. The relative reactivities of hydroxyl groups for mixtures of Me alpha- and Me beta-Galf were HO-2 > HO-3 > HO-6 > HO-5, that of Me alpha and Me beta-Araf HO-2 > HO-3 > HO-5, and that of Me alpha-Araf HO-2 > HO-3 > HO-5