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Aqueous formamide solutions can promote catalytic amidolysis of an activated organophosphate in a regiospecific process.

Pain is a common and distressing symptom of many diseases and its clinical treatment generally involves analgesics and anti-inflammatory drugs. This study evaluated the toxicity of Ilex paraguariensis A. St.-Hil. (Aquifoliaceae) aqueous extract (leaves, petioles and branches) and its performance in a nociceptive response. Hepatotoxicity, psycho-stimulant test and evaluation of enzyme markers for liver damage were also tested. Chromatographic analysis by UPLC-MS demonstrated a series of isomeric monocaffeoylquinic acids, isomers of dicaffeoylquinic acid, flavonol glycosides, and saponins. Phase I and II of nociception were obtained for meloxicam, dexamethasone and aqueous Ilex paraguariensis extract. Ilex paraguariensis extract concentration was negatively correlated (R = –0.887) with alanine aminotransferase (p < 0.05) in acetaminophen-induced hepatotoxicity test, indicating hepatoprotective activity of this extract. Ilex paraguariensis extract also presented analgesic properties equivalent to drugs that already have proven efficacy. Notably, the administration of multiple doses of Ilex paraguariensis extract was considered safe from the therapeutic point of view.

Successful long-term cryopreservation of lepidopteran eggs for the mass production of parasitoids of the genus Trichogramma Westwood (Hymenoptera: Trichogrammatidae) requires a more tolerant species to withstand ultra-low temperatures. We compared the viability of eggs of Anticarsia gemmatalis Hubner (Lepidoptera: Erebidae) and Mythimna sequax Franclemont (Lepidoptera: Noctuidae), cryopreserved in liquid nitrogen, and evaluated the viability of M. sequax eggs to support the large-scale production of high-quality Trichogramma species. The presence of anti-freezing metabolites in lepidopteran eggs was detected and quantified by 1H nuclear magnetic resonance (NMR) spectroscopy. Parasitism by Trichogramma pretiosum Riley of M. sequax eggs stored for 30 days was significantly higher (84.2%) in comparison to A. gemmatalis (6.7%). The 1H NMR spectrum showed that the amounts of anti-freezing metabolites maltodextrin and trehalose were greater in M. sequax eggs, a possible explanation for their suitability to cryopreservation in comparison to A. gemmatalis. Eggs of M. sequax cryopreserved for up to 12 months maintained a mean rate of parasitism between 80 and 90% by Trichogramma atopovirilia Oatman and Platner and T. pretiosum, the latter declining to 79% after 12 months. Both Trichogramma species can exploit long-term storage of M. sequax eggs at ultra-low temperatures, without any apparent fitness penalty to the first generation. This is the first record of eggs of a noctuid species being cryopreserved for up to one year and remaining susceptible to parasitism by Trichogramma species. This procedure should enable large-scale production of hosts with high nutritional quality, and the production of parasitoids that is synchronized better with their use as biological control agents in annual crops.

Fructan fatty acid esters are amphiphilic compounds with potential applications in the food industry due to their surface-active properties on the air/water interface. In this work, the regioselect...

Biodegradable films are an excellent alternative to using non-biodegradable materials. Plant extracts are rich in antioxidant and antimicrobial components and can be used as components in active packaging to improve food preservation. This study demonstrated that the ethyl acetate fraction of the crude bark extract of Euphorbia umbellata presented high amounts of phenolic compounds (515.81 ± 29.61 mg of galic acid per gram of fraction) with concentration-dependent antioxidant potential (DPPH•—IC50 1.95 ± 0.01 μg/ml; ABTS•+—IC50 1.40 ± 0.15 µg/ml; FRAP–2,680.05 ± 0.02 µmol of trolox per gram of fraction), and antimicrobial activities against Staphylococcus aureus and Escherichia coli (inhibition halo diameter larger than 8 mm). The addition of this fraction into biodegradable starch-based films allowed the formation of an active matrix, with minor mechanical changes when the ethyl acetate fraction was used at 2.5 and 5.0% (w/w), registering tensile strength results of 29.5 ± 4.0 MPa for inert film; 29.5 ± 4.0 MPa for 2.5% (w/w) of E. umbellata ethyl acetate extract, and 25.1 ± 1.9 MPa for 5% (w/w) of E. umbellata ethyl acetate extract. A heterogeneous microstructure in the presence of ethyl acetate extract resulted in lack of homogeneity and rough surface of the films, maintaining important antioxidant and antibacterial activities. This material can be considered as an alternative for production and application as biodegradable and active food packaging.

Organophosphate (OP) pesticides are responsible for numerous human deaths every year. Nucleophilic substitution is an important method to mitigate the toxicity of obsolete stocks of OPs. Herein, the degradation of O,O-diethyl-2,4-dinitrophenyl phosphate (DEDNPP) and pesticide diethyl-4-nitrophenyl phosphate (Paraoxon) promoted by 1,2,4-triazole (TAZ) was investigated by means of kinetic studies, nuclear magnetic resonance (NMR) analyses, and theoretical calculations. Results showed fast degradation of OPs is promoted by the anionic form of the nucleophile (TAZ(-)) in pH > 8.5 (optimal at pH = 11). Rate enhancements of 106 and 105-fold in relation to neutral hydrolysis of DEDNPP and Paraoxon were observed, respectively, consistent with alpha-nucleophiles reactivity. TAZ(-) regioselectively promotes the degradation of DEDNPP via P-O bond break, forming a quickly hydrolyzable phosphorylated intermediate, regenerating the nucleophile. Calculations using M06-2X/6-311++G(d,p) level of theory revealed that the equivalent nitrogen atoms of TAZ(-) are the main nucleophilic center of the molecule. This study expands the knowledge on the reactivity of iminic compounds as detoxificant agents of OPs, indicating the efficiency and selectivity of TAZ(-) in aqueous medium, encouraging the design of novel TAZ-based catalysts.

Cereus hildmannianus is a cactus exhibiting morphological and physiological adaptation of its cladodes which ensuring growth in climatic and soil conditions unfavourable for many plant species. Currently, limited water resources and increasing demand for renewable energy make cacti a biomass source for the production of biofuels. Somaclones regenerated from callus in vitro can be a source of new raw material in useful plants. The objective of this work was to determine if the regenerated plants showing two different morphologies present polysaccharide composition different from the wild plant. Somaclones aqueous extraction shows the absence of soluble polysaccharides as mucilage. The alkaline extraction of in vivo cultivated plant showed the presence of starch, type I arabinogalactan, and arabinoxylan and the somaclones showed type I arabinogalactan and arabinoxylan in both morphologies. Hemicelluloses found in the somaclones are not different from in vivo cultivated plants, but somaclones not almost biosynthesize mucilage and starch.

The seed oil of Annona salzmannii A. DC. was analyzed by GC-MS and 1H qNMR, revealing a mixture of unsaturated (80.5%) and saturated (18.7%) fatty acids. Linoleic (45.3%) and oleic (33.5%) acid were the major unsaturated fatty acids identified, while palmitic acid (14.3%) was the major saturated fatty acid. The larvicidal effects of A. salzmannii seed oil were evaluated against third-instar larvae of Aedes aegypti (Linn.). The oil exhibited moderate larvicidal activity, with a LC50 of 569.77 ppm (95% CI = 408.11 to 825.88 ppm). However, when the cytotoxic effects of the oil were evaluated, no expressive antiproliferative effects were observed in tumor cell lines B16-F10 (mouse melanoma), HepG2 (human hepatocellular carcinoma), K562 (human chronic myelocytic leukemia), HL-60 (human promyelocytic leukemia), and non-tumor cell line PBMC (peripheral blood mononuclear cells), with IC50 values > 50 μg·mL-1. This is the first study to evaluate the chemical composition, larvicidal and cytotoxic activity of A. salzmannii seed oil.

Oxidative stress is the cause of numerous diseases in humans, therefore, there has been a continuous search for novel antioxidant molecules. Fungal chitosan is an attractive molecule that has several applications (antifungal, antibacterial, anticancer and antiparasitic action) owing to its unique characteristics, however, it exhibits low antioxidant activity. The aim of this study was to obtain fungal chitosan (Chit-F) from the fungus Rhizopus arrhizus and synthesize its derivative, fungal chitosan-gallic acid (Chit-FGal), as a novel antioxidant chitosan derivative for biomedical use. A low molecular weight Chi-F (~3.0 kDa) with a degree of deacetylation of 86% was obtained from this fungus. Chit-FGal (3.0 kDa) was synthesized by an efficient free radical-mediated method using hydrogen peroxide (H2O2) and ascorbic acid. Both Chit-F and Chit-FGal showed similar copper chelating activities, however, Chit-FGal was more efficient as an antioxidant, exhibiting twice the total antioxidant capacity than Chi-F (p &lt, 0.05). Furthermore, H2O2 (0.06 M) promoted a 50% decrease in the viabilities of the 3T3 fibroblast cells. However, this effect was abolished in the presence of Chit-FGal (0.05–0.25 mg/mL), indicating that Chit-FGal protected the cells from oxidative damage. These results suggest that Chit-FGal may be a promising agent to combat oxidative stress.

Polysaccharides α-D-galactan (GAL-Am) and β-D-glucan (GLC-Am) were obtained from Amanita muscaria fruiting bodies. They were purified using different methodologies, such as Fehling precipitation (for both fractions), freeze-thawing process and ultrafiltration (for GLC-Am). Results showed that the GAL-Am has (1 → 6)-linked Galp main chain branched at O-2 by terminal Galp units and has not been previously reported. Besides, GLC-Am has (1 → 3)-linked Glcp in the main chain, substituted at O-6 by (1 → 6)-linked β-Glcp units. Both are water-soluble, with 9.0 × 103 g/moL and 1.3 × 105 g/moL, respectively. GAL-Am and GLC-Am presented a selective proliferation reduction against B16-F10 melanoma cell line, not affecting non tumoral BALB/3T3 fibroblast cell line. Furthermore, both fractions reduced clonogenic capacity of melanoma cell line over an extended period of time. These results were obtained without modulations in B16-F10 cell adhesion, reinforcing the biological activities towards cell proliferation impairment and eliciting these polysaccharides as promising compounds to further exploration of their antimelanoma properties.

Pain is a common and distressing symptom of many diseases and its clinical treatment generally involves analgesics and anti-inflammatory drugs. This study evaluated the toxicity of Ilex paraguariensis A. St.-Hil. (Aquifoliaceae) aqueous extract (leaves, petioles and branches) and its performance in nociceptive response. Hepatotoxicity, psychostimulant test and evaluation of enzyme markers for liver damage were also tested. Chromatographic analysis by UPLC-MS demonstrated a series of isomeric monocaffeoylquinic acids, isomers of dicaffeoylquinic acids, flavonol glycosides, and saponins. Phase I and II of nociception were obtained for meloxicam, dexamethasone and aqueous Ilex paraguariensis extract. Ilex paraguariensis extract concentration was negatively correlated (R = –0.887) with alanine aminotransferase (p < 0.05) in acetaminophen-induced hepatotoxicity test, indicating an hepatoprotective activity of this extract. Ilex paraguariensis extract also presented analgesic properties equivalent to drugs that already have proven efficacy. Notably, administration of multiple doses of Ilex paraguariensis extract was considered safe from the therapeutic point of view.

A 12.4 kDa laminarin (LM) composed of &beta, (1&rarr, 3)-glucan with &beta, 6)-branches was extracted from brown seaweed Lobophora variegata and modified via carboxylation using dielectric barrier discharge (LMC), conjugation with gallic acid (LMG), and sulfation (LMS). Analyses of the chemical composition of LMC, LMG, and LMS yielded 11.7% carboxyl groups, 1.5% gallic acid, and 1.4% sulfate content, respectively. Antioxidant activities of native and modified laminarins were assessed using six different in vitro methods. Sulfation stopped the antioxidant activities of LM. On the other hand, carboxylation improved cooper chelation (1.2 times). LMG was found to be a more efficient antioxidant agent than LM in terms of copper chelation (1.3 times), reducing power (1.3 times), and total antioxidant capacity (80 times). Gallic acid conjugation was further confirmed using Fourier transform infrared spectroscopy (FT-IR) and one- and two-dimensional NMR spectroscopy analyses. LMG also did not induce cell death or affect the cell cycle of Madin&ndash, Darby canine kidney (MDCK) cells. On the contrary, LMG protected MDCK cells from H2O2-induced oxidative damage. Taken together, these results show that LMG has the potent antioxidant capacity, and, therefore, potential applications in pharmacological and functional food products.

Oils enriched with diacylglycerol have received a great deal of attention due to their potential health benefits when used as a replacement for conventional edible oils. This work reports experimental results and thermodynamic modeling of liquid-liquid equilibrium of systems containing mono-, di- and triacylclycerols from olive oil, glycerol and isopropanol. The UNIQUAC model was used in the description of phase equilibria, which was well fitted to experimental data with root mean square deviations equal or lower then 1.4 wt% for the all studies systems. The results presented in this work show the feasibility of diacylglycerol separation from other acylglycerols through liquid-liquid extraction using glycerol and isopropanol as solvents.

Background and Aims Uremic toxins (UTs) accumulate in the circulation of patients with chronic kidney disease (CKD), contributing to poor outcomes. One of the main objectives of renal replacement therapies is the removal of UTs, and while HD is restricted to the clearance of low molecular weight UTs, hemodiafiltration (HDF) adds efficiency to the removal of medium molecular weight UTs through the association of diffusion and convection methods. Metabolomic analysis from nuclear magnetic resonance (NMR) spectra offers efficient data about metabolites profile, indicating the main features of HDF, and possibly highlighting the positive effects of this modality. Thus, the aim of the present study is to evaluate the profile of metabolites in the circulation of a cohort of patients undergoing HDF. Method From 97 individuals participating in the HDF arm of a randomized control trial (ClinicalTrials.gov Identifier: NCT02787161), we randomly (using Python language; NumPy library) selected 6 patients (mean age 49, 6 males, 44% diabetics, average duration of 245 minutes, blood flow of 400 mL/min, target convective volume of 22 L per treatment). Patient’s serum samples were collected in the beginning of the study (baseline) and 6 months. The samples were prepared with deuterium oxide, sodium trimethylsilylpropanesulfonate as internal standard (ð=0.0 ppm), and further submitted to 1H NMR spectra recorded with Bruker 600 MHz AVANCE III spectrometer in order to identify the patient’s metabolomic profile. Data was evaluated with t-test and results were then compared with the Human Metabolome Database (HMDB) for descending order of the Jaccard similarity score. Results A total of 50 metabolites were investigated. Interestingly, 2 metabolites were significantly (P Conclusion We observed a reduction of acetate and dimethylglyoxal levels after six months of HDF treatment, in comparison to baseline, most likely as a consequence of the increase in clearance of these molecules. Hypothetically these changes in solute removal could explain the positive effects of HDF on clinical outcomes, and further studies should address this potential benefit of the modality.

Under conditions of carbon starvation or thermal, osmotic, or oxidative shock, mutants affected in the synthesis or mobilization of poly-3-hydroxybutyrate (PHB) are known to survive less well. It is still unclear if the synthesis and accumulation of PHB are sufficient to protect bacteria against stress conditions or if the stored PHB has to be mobilized. Here, we demonstrated that mobilization of PHB in Herbaspirillum seropedicae SmR1 was heat-shock activated at 45°C. In situ proton ((1)H) nuclear magnetic resonance spectroscopy (i.e., (1)H-nuclear magnetic resonance) showed that heat shock increased amounts of 3-hydroxybutyrate (3HB) only in H. seropedicae strains able to synthesize and mobilize PHB. H. seropedicae SmR1 mutants unable to synthesize or mobilize PHB were more susceptible to heat shock and survived less well than the parental strain. When 100 mM 3-hydroxybutyrate was added to the medium, the ΔphaC1 strain (an H. seropedicae mutant unable to synthesize PHB) and the double mutant with deletion of both phaZ1 and phaZ2 (i.e., ΔphaZ1.2) (unable to mobilize PHB) showed partial rescue of heat adaptability (from 0% survival without 3HB to 40% of the initial viable population). Addition of 200 mM 3HB before the imposition of heat shock reduced protein aggregation to 15% in the ΔphaC1 mutant and 12% in the ΔphaZ1.2 mutant. We conclude that H. seropedicae SmR1 is naturally protected by 3HB released by PHB mobilization, while mutants unable to generate large amounts of 3HB under heat shock conditions are less able to cope with heat damage. IMPORTANCE Bacteria are subject to abrupt changes in environmental conditions affecting their growth, requiring rapid adaptation. Increasing the concentration of some metabolites can protect bacteria from hostile conditions that lead to protein denaturation and precipitation, as well as damage to plasma membranes. In this work, we demonstrated that under thermal shock, the bacterium Herbaspirillum seropedicae depolymerized its intracellular stock polymer known as poly-3-hydroxybutyrate (PHB), rapidly increasing the concentration of 3-hydroxybutyrate (3HB) and decreasing protein precipitation by thermal denaturation. Mutant H. seropedicae strains unable to produce or depolymerize PHB suffered irreparable damage during thermal shock, resulting in fast death when incubated at 45°C. Our results will contribute to the development of bacteria better adapted to high temperatures found either in natural conditions or in industrial processes. In the case of H. seropedicae and other bacteria that interact beneficially with plants, the understanding of PHB metabolism can be decisive for the development of more-competitive strains and their application as biofertilizers in agriculture.

The current study was carried out by a bioguided fractionation of a hexane extract of the latex of Euphorbia umbellata against leukemic cells. Samples were analyzed by NMR, GC/MS, triterpenes quantification, and MTT reduction assay. Morphological, cell cycle, mitochondrial membrane potential and caspases 3/7 analyses were performed for the dichloromethane and ethanol fractions, and selectivity index for the dichloromethane fraction. NMR analysis presented characteristic signals of terpenes and steroids, data were confirmed by the quantification of triterpenes and GC/MS analysis. MTT reduction assay demonstrated that HL-60 was the most sensitive cell lineage against dichloromethane and ethanol fractions. Compounds of these matrices caused morphological changes compatible with apoptosis induction, altered cell cycle, increment of depolarized population cells and activation of caspases 3/7. Selectivity indices were higher than 22.44. Bioguided-fractionation study showed that samples of the latex of E. umbellata raised the activity of the phytocomplex against leukemic cells, and the cytotoxicity can be associated with an apoptosis pathway.

Ethnopharmacological relevance The species Euphorbia umbellata (leitosinha) has been traditionally used for the treatment of inflammatory diseases and cancer. Aim of the study: Evaluation the effect of E. umbellata latex extracts obtained with hexane, chloroform, ethyl acetate and methanol on the activation of the complement pathways and neutrophil chemotaxis. Materials and methods The latex was partitioned using Soxhlet apparatus and hexane, chloroform, ethyl acetate and methanol as solvents. The classical and alternative pathway activity were performed by hemolytic assays with sensitized sheep or rabbit erythrocytes, respectively; the lectin pathway activity was quantified by ELISA, through the measurement of C4 molecules and the chemotaxis of human neutrophils was performed using 1% casein as the chemotactic inducer and Boyden's chamber. GC-Q-ToF and NMR analyses were applied to evaluate the chemical composition of E. umbellata latex extracts. Results All E. umbellata latex extracts exhibited an inhibitory effect on the activation of the alternative pathway. Methanol and ethyl acetate extracts inhibited the classical pathway while chloroform extract activated this pathway. Ethyl acetate and hexane extracts inhibited lectin activation. All E. umbellata extracts inhibited casein-induced neutrophil chemotaxis. Terpenes and phenolic compounds have been suggested to be present in the E. umbellta latex extracts. Conclusion The E. umbellata latex was able to modulate the functions of the immune system. Thus, it is possible to infer that the terpenes and phenolic compounds of the phytocomplex of E. umbellata latex can contribute for the activity on the complement pathways.

Uremic toxins (UTs) accumulate in the circulation of patients with chronic kidney disease (CKD). High volume hemodiafiltration (HDF) improves clearance of low and medium molecular weight UTs compared to HD. The present study is a post-hoc analysis comparing the metabolomic profile in serum from patients under high flux HD (hf-HD) and HDF in HDFIT, a multicentric randomized controlled trial (RCTs). Per protocol, serum samples were collected pre- and post- dialysis treatments at randomization (baseline) and at the end of the follow up (6 months) and stored in a biorepository. Random (pre- and post-dialysis) samples from nine patients in study arm were selected at baseline and at the end of the follow up. To compare the samples, 26 possibly matching metabolites were identified by a t-test among the four groups using 1H nuclear magnetic resonance (NMR). To evaluate the comparison between the modalities is a single treatment session, the clearance rates (CRs) of each metabolite were calculated based on pre-dialysis and post-dialysis samples. In addition, to evaluate to effect of UT removal during the trial follow up period, the pre-dialysis metabolite concentrations at the baseline and at 6 months were compared among the two arms of the study. There was no significant difference between in the single session CRs of metabolites when hf-HD and HDF were compared. On the other hand, the comparison between baseline and 6-month (long-term evolution) led to the identification of 16 metabolites that differentiated the hf-HD and the HDF evolutions. Most of these 16 metabolites are involved in several important metabolic pathways, such as metabolism of phenylalanine and biosynthesis of phenylalanine, tyrosine, and tryptophan, which are related to UTs and cardiovascular disease development. Although no difference was observed between hf-HD and HDF samples before and after a single session, concentrations of CKD-relevant metabolites and associated pathologies were stable in the HDF samples, but not in the hf-HD samples, over the six-month period, suggesting that HDF enhances long-term stability.

Process optimization for producing diacylglycerol-enriched oils has received great attention due the importance of these products for healthier and more balanced diets. This work reports experimental data of liquid-liquid phase equilibrium of hydrolyzed olive oil (which comprises on its composition triacylglycerols, diacylglycerols, monoacylglycerols and free fatty acids) with ethanol and water, and subsequent analysis of oxidative stability of the oil obtained from the liquid phases. Liquid-liquid equilibrium experimental data obtained were modeled using the UNIQUAC thermodynamic model, which was well fitted to experimental data with a root mean square deviation of 1.76 wt%. The results obtained demonstrated the potentiality of liquid-liquid extraction as a suitable procedure for olive oil enrichment in diacylglycerol, after a hydrolysis procedure. It was also observed that liquid-liquid phase separation has a significant effect on the content of important antioxidants present in the olive oil, what affects the oxidative stability of the samples submitted to the liquid phase extraction. From the results obtained in this study, it has been also noticed that the acylglycerols and free fatty acids profile in of the oil samples presented a major rule on their stabilities.

Three polysaccharide fractions were isolated from blackberry wine. The crude extract BWPs was obtained with ethanol precipitation and freeze-thawing process, it was then submitted to Fehling treatment, giving soluble BWPFs and insoluble BWPFp fractions. These fractions were characterized by Gas Chromatography-Mass Spectrometry (GC–MS) and Nuclear Magnetic Resonance (NMR). Major polysaccharides were identified for each fraction: mannan, type II arabinogalactan and type I rhamnogalacturonan for BWPs, a mannan formed by a major chain of α-Man p (1 → 6)-linked units, O -2 substituted with α- d -Man p (1 → 2)-linked side chains for BWPFp and a AG II formed by a major chain of β - d -Gal p (1 → 3)-linked, substituted at O -6 by side chains of the β - d -Gal p (1 → 6)-linked, which then are substituted at O -3 by non-reducing units of α - l -Ara f and a RG I, formed by [→4)- α - d -Gal p A-(1 → 2)- α - l -Rha p -(1→] n for BWPFs. Anti-inflammatory effects of polysaccharide fractions were evaluated in RAW 264.7 cells. Fractions markedly reduced nitric oxide (NO) and pro-inflammatory cytokine production (TNF-α and IL-1β) in LPS-treated cells.

The structural characterization of the polysaccharides and in vitro anti-inflammatory properties of Cabernet Franc (WCF), Cabernet Sauvignon (WCS) and Sauvignon Blanc (WSB) wines were studied for the first time in this work. The polysaccharides of wines gave rise to three fractions of polysaccharides, namely (WCF) 0.16%, (WCS) 0.05% and (WSB) 0.02%; the highest one was chosen for isolation of polysaccharides (WCF). It was identified the presence of mannan, formed by a sequence of α- d -Manp (1 → 6)-linked and side chains O-2 substituted for α- d -mannan (1 → 2)-linked; type II arabinogalactan, formed by (1 → 3)-linked β- d -Galp main chain, substituted at O-6 by (1 → 6)-linked β- d -Galp side chains, and nonreducing end-units of arabinose 3-O-substituted; type I rhamnogalacturonan formed by repeating (1 → 4)-α- d -GalpA-(1 → 2)-α-L-Rhap groups; and traces of type II rhamnogalacturonan. The polysaccharide mixture and isolated fractions inhibited the production of inflammatory cytokines (TNF-α and IL-1β) and mediator (NO) in RAW 264.7 cells stimulated with LPS.

The occurrence of a natural and unmodified highly sulfated chondroitin sulfate from Litopenaeus vannamei heads (sCS) is herein reported. Its partial digestion by Chondroitinases AC and ABC together with its electrophoretic migration profile revealed it as a highly sulfated chondroitin sulfate despite its average molecular weight being similar to CSA. Using orthogonal 1D/2D NMR experiments, the anomeric signals (δ 4.62/106.0) corresponding to unusual 2,3-di-O-Sulfo-GlcA (∼36%), U33S (δ 4.42/84.1, ∼63%) and U22S (4.12/80.1, ∼50%) substitutions were confirmed. In addition, non-sulfated GlcA (δ 4.5/106.3) linked to 4-O- (A14S, 36%) or 6-O-Sulfo (A16S, 28%) GalNAc (δ 4.64/103.5) was observed. Although the biological role of sCS in shrimp is unknown, its influence on hemostasis was also demonstrated. The sCS identification brings to light new questions about the hierarchical model of GAGs biosynthesis and contributes to the better understanding of the subtle relationship between GAGs structure and function.

Knowledge about lignin structure after isolation is paramount for demonstrating its potential for industrial applications. This paper presents for the first time the detailed characterization of LignoForce™ hardwood (LFHL) and softwood (LFSL) kraft lignins that were obtained from industrially realistic lignin production facilities. The moisture content was circa 8 % for both LFHL and LFSL, and their total carbon contents were 62.8 and 64.4 %, respectively. Both samples had low ash and carbohydrate contents and total lignin contents higher than 97 %. The macromolecular properties and functional groups of lignin were characterized by nuclear magnetic resonance, analytical pyrolysis (Py-GC–MS), thermal analysis, mass spectrometry, and size exclusion chromatography. LFHL presented lower aliphatic and phenolic OH groups, higher methoxyl content (S/G ratio of 1.97), lower weight-average apparent molecular mass (Mw) and degree of condensation, higher solubility in organic solvents, and lower thermal stability compared to LFSL. C1-Guaiacol and C1-syringol predominated in the Py-GC–MS analyses of LFSL and LFHL, respectively, whose β-O-4′ contents were 5.8 and 6.2 per 100 aromatic units. The high total OH groups of both lignins suggests their suitability for replacing petroleum-based polyols in the preparation of polyurethane foams. LFHL would be preferable in this application due to its higher solubility in the reaction medium. Also, the lower Mw and glass transition temperature of LFHL would facilitate its blending with polymers in thermoplastic applications. LFSL would be a suitable replacement of phenol in phenol-formaldehyde resins, and its higher Mw and improved thermal stability are expected to be useful for making carbon fibers.

Melanised cell walls and extracellular polymeric matrices protect rock-inhabiting microcolonial fungi from hostile environmental conditions. How extracellular polymeric substances (EPS) perform this protective role was investigated by following development of the model microcolonial black fungus Knufia petricola A95 grown as a sub-aerial biofilm. Extracellular substances were extracted with NaOH/formaldehyde and the structures of two excreted polymers studied by methylation as well as NMR analyses. The main polysaccharide (~ 80%) was pullulan, also known as α-1,4-; α-1,6-glucan, with different degrees of polymerisation. Αlpha-(1,4)-linked-Glcp and α-(1,6)-linked-Glcp were present in the molar ratios of 2:1. A branched galactofuromannan with an α-(1,2)-linked Manp main chain and a β-(1,6)-linked Galf side chain formed a minor fraction (~ 20%). To further understand the roles of EPS in the weathering of minerals and rocks, viscosity along with corrosive properties were studied using atomic force microscopy (AFM). The kinetic viscosity of extracellular K. petricola A95 polysaccharides (≈ 0.97 × 10−6 m2 s−1) ranged from the equivalent of 2% (w/v) to 5% glycerine, and could thus profoundly affect diffusion-dominated processes. The corrosive nature of rock-inhabiting fungal EPS was also demonstrated by its effects on the aluminium coating of the AFM cantilever and the silicon layer below.

Several bioactive sulfated galactans have been isolated from the tunic of different species of ascidians. The biological activity of this kind of polysaccharides has been related with the presence and position of sulfate groups, and by the chemical composition of this kind of polysaccharides. A sulfated galactan (1000RS) was isolated from the tunic of the Brazilian ascidia Microcosmus exasperatus through proteolytic digestion, ethanol precipitation, dialysis and freeze-thaw cycles. Homogeneity and molecular weight were estimated by using size exclusion chromatography. Monosaccharide composition and type of linkage were assessed by Gas chromatography coupled to mass spectrometry and the sulfate content was quantified through gelatin/BaCl2 method. These experiments along with NMR and FTIR analysis allowed to claim that the galactan backbone is mainly composed of 4-linked α- l -Galp units. In addition, they permitted to establish that some of the galactose residues are sulfated at the 3-position. This sulfated polysaccharide, which has an average molecular mass of 439.5 kDa, presents anticoagulant effect in a dose-dependent manner through the inhibition of the intrinsic coagulation pathway.

This study reports the presence of mannogalactans in Pholiota nameko and Pleurotus eryngii edible mushrooms, with their purification and detailed structural characterization. Two mannogalactans (R30PN and E30PN) were reported in P. nameko, differing mainly in their Mw (37.9 × 104 g/mol and 2.4 × 104 g/mol). The mannogalactans presented a linear (1→6)-α- d -galactan in the main chain, and single branches at O-2 by β- d -Manp as non-reducing end units. The main chain also contained naturally 3-O-methylated Galp units, in amounts that varied from 15.2–23.7 %. P. nameko mannogalactans were described for the first time. Heteropolysaccharides from mushrooms may display functional activities, like antitumor, anti-inflammatory and immunostimulatory which are related to their chemical structure. Thus, this study is of relevance since it unveils the structural details of two mannogalactans from P. nameko and one from P. eryngii mushrooms. Further studies are encouraged to verify how these structural differences may interfere in their biological activities.

In this study, we isolated and structurally characterized, for the first time, a galactoglucomannan (GGM) from the pulp of gabiroba, a Myrtaceae family species. The HPSEC-MALLS-RI analysis showed a homogeneous polysaccharide with molar mass of 25,340gmol-1. The monosaccharide composition showed that the GGM consisted of Man:Glc:Gal in a molar ratio of 1:1:0.6. Methylation and 1D and 2D NMR analyses suggested that the main chain of the GGM consisted of β-d-Glcp and β-d-Manp units (1→4)-linked. The α-d-Galp substitutions occur mainly at O-6 position of β-d-Manp units. The glycosidic linkages of the GGM were evident by the presence of the characteristic signals of 4-O-substituted residues at δ 78.6/3.69 for both β-d-Glcp and β-d-Manp. Furthermore, the O-6 substitutions for both β-d-Glcp and β-d-Manp units were confirmed by signals at δ 67.1/4.00 and 3.93. The interglycosidic correlations, obtained through the analysis of the HMBC spectrum, further confirm the structure.

Croton cajucara Benth. is a medicinal plant popularly used in the Brazilian Amazonia, where it is known as sacaca, being consumed as tea, decoction or infusion of the leaves and stem bark. From a decoction of the leaves, a comprehensive phytochemical analysis was developed by liquid chromatography-mass spectrometry. Many compounds were identified for the first time in C. cajucara, such as O-glycosides of kaempferol and quercetin, flavonoid-C-glycosides, tannins and cinnamic acid derivatives. These compounds were fractionated by polarity and assayed for their anti-inflammatory activity, using a model of mice edema, induced by an intraplantar injection of carrageenan. All fractions exhibited anti-inflammatory properties.

Background Advanced glycation end products (AGEs) have been related to the pathogenesis of cardiovascular diseases (CVD), chronic kidney disease (CKD) and diabetes mellitus. We sought to investigate the binding capacity of sevelamer to both AGEs and uremic serum in vitro and then test this pharmaceutical effect as a potential vascular anti-inflammatory strategy. Methods AGEs were prepared by albumin glycation and characterized by absorbance and electrophoresis. Human endothelial cells were incubated in culture media containing AGEs and uremic serum with or without sevelamer. Receptor for advanced glycation end product (RAGE) expression was evaluated through immunocytochemistry and western blot to explore the interactions between AGEs and the endothelium. Inflammatory and endothelial dysfunction biomarkers, such as interleukin 6 (IL-6) and IL-8, monocyte chemoattractant protein-1 (MCP-1), plasminogen activator inhibitor-1 (PAI-1) and serum amyloid A (SAA) were also measured in cell supernatant. The chemotactic property of the supernatant was evaluated. Results AGEs significantly induced the expression of RAGE, inflammatory and endothelial activation biomarkers [IL-6, (P

Mucilage extracted from leaves of Pereskia aculeata, popularly called Barbados gooseberry (or ora-pro-nobis in Brazil), was investigated in terms of structural, chemical and physical-chemical properties. The mucilage was obtained with a new method using water at room temperature. Analysis of polysaccharide by GC-MS and NMR techniques showed the presence of galactose, arabinose, rhamnose, fucose and partially esterified galacturonic acid, whose main chain consisted of β-d-galactose linked (1 → 4), partially substituted at C-3 by arabinose, making it possible to characterize an arabinogalactan type I, highlighting the presence of fucose units. In water saline solution, the material is highly heterogeneous, as observed by size exclusion chromatography, where the main fraction is dispersed (Đ = 1.3) with weight average molar mass (Mw) of 7.9 × 105 g/mol. In addition, the intrinsic viscosity [η] (26 dL/g), by the Mark-Houwink equation, and the [η] dependence of Mw indicate that the mucilage has random conformation and flexible chains in sodium nitrate solution, with a radius of gyration of 41 nm and critical concentration (c*) of 4.6 mg/mL. Tensiometry allowed determining the critical adsorption concentration (CCA) and the critical self-association concentration (CSAC), as 1 and 6 mg/mL, respectively, and the sample showed mild surfactant characteristics. In aqueous dispersion (30 g/L), the rheological measurement showed pseudoplastic behavior of the mucilage, with polyelectrolyte characteristics. Finally, the pre-heating of the mucilage at 70 °C for 10 min caused an increase in viscosity and improved pseudoplasticity, for better interaction and molecular solubilization after cooling.

This work reports experimental kinetic data in addition to conventional and modified mathematical models for predicting free fatty acid production from refined and waste frying soybean oils in subcritical water. The experiments were performed in batch mode until 220 min of reaction, at an oil to water mass ratio of 1:4 and temperatures of 250 and 275 °C. The proposed modified kinetic model presented a good agreement with the experimental data, with a mean absolute error of 2.17% between experimental data of free fatty acid yields and calculated values (the conventional model presented an error of 2.41%). The experimental data used in the model parameter estimation were obtained from the hydrolysis of refined oil and frying oil used on a single frying procedure. The modified kinetic model was also used to predict the hydrolysis of a mixture of waste frying oils submitted to more than one cycle of frying (results not used for parameter estimation), and a mean error of 3.04% was observed when comparing the calculated values to experimental data (the conventional model presented an error of 4.98%). It was observed that the effect of frying is positive and statistically significant in the hydrolysis kinetics, which can justify the use of the proposed modified model for predicting yields of free fatty acids from oils submitted to different home frying cycles.

Cell wall polysaccharides from filamentous fungi and yeasts have been reported as antioxidant and antiproliferative polymers. Thus, we evaluated these activities from cell wall polysaccharides from Kluyveromyces marxianus CCT7735. By using a centrifugal filter, a 203 kDa α-d-mannan (KMM-5) was obtained. KMM-5 exhibited no effect on HeLa cells and a weak antiproliferative activity against Hep-G2 cells. In addition, at higher concentrations, it presented a cytotoxicity to the normal cell line, 3T3. However, KMM-5 showed copper- and iron-chelating abilities, the latter of which presented improved activity. By using 2D-NMR COSY, HSQC edited and HMBC experiments, a structure arrangement was proposed. The main chain was formed by 6)-α-d-Manp-(1 → 6) units substituted at the 2-O-position by non-reducing terminals α-d-Manp-(1 → 2) and by a branched tetrasaccharide. The latter was formed by an internal 2)-α-d-Manp-(1 → 2) unit with linked to it a 2,3)-α-d-Manp-(1 → 2) unit substituted at the 2-O-position by a non-reducing terminal α-d-Manp-(1 → 2), and at the 3-O-position by a non-reducing terminal α-d-Manp-(1 → 3). In conclusion, we considered K. marxianus CCT7735 a source of natural and renewable polysaccharides with pharmacological properties.

Croton cajucara Benth. is a tree from the Amazon Forest, where it is known as sacaca. Its leaves and barks are used in medicinal preparations to treat different diseases, including gastric ulcers. The crude polysaccharide fraction (CCP), obtained from the hot aqueous extract of C. cajucara leaves, was able to promote gastroprotection on an ethanol induced gastric ulcer model. Therefore, a bioguided fractionation was performed to isolate the active polysaccharide fraction. After freezing-thawing, ultrafiltration and dialyses at 100, 50, and 25kDa cut-off membranes, fraction 25R was obtained. It contained glucose, galactose, rhamnose, arabinose, galacturonic acid and mannose in a 7:5:5:3:1:1 molar ratio approximately, and had a Mw of 42,840g/mol. Methylation analysis and NMR spectroscopy indicated that 25R is a very complex polysaccharide fraction containing type I rhamnogalacturonan, arabinan, type I arabinogalactan, type II arabinogalactan, rhamnan, starch and mannan. It was able to reduce ethanol-induced gastric ulcers in rats, through preservation of mucus and GSH levels.

Hyaluronic acid (HA) and galactomannans (such as guar gum - GG and locust bean gum - LBG) are biomacromolecules with various applications due to their physicochemical characteristics in solution, especially their ability to form films or hydrogels in some conditions. Some of these applications are possible because they are nontoxic and biocompatible. Mixtures of HA and GG or LBG, at pH 7.4, were prepared at 25 g·L−1 of HA:LBG or HA:GG with percentages of 50:50, 60:40, 70:30 and 80:20 (w/w) and evaluated by rheology and NMR experiments regarding the interactions. The best synergism observed was between HA:LBG, most pronounced at 50% (w/w), where the dynamic modulus values G′ and G″ indicated the formation of physical gels with viscoelastic behavior close to that of HA. 2D NOESY spectra revealed hydrogen interconnectivity of H-1 glucuronic acid units of HA with H-5 mannose units of the main galactomannan chains. This synergism, besides producing a hydrogel with interesting properties, also can decrease the cost by partially replacing HA in many applications, such as in nutraceutical foods, support for tissue engineering and cosmetics.

A novel derivative of dextran, dextran&ndash, gallic acid (Dex&ndash, Gal), obtained from simple conjugation with gallic acid, was synthesized by an efficient free radical-mediated method. To verify the synthesis of Dex&ndash, Gal, 1H-nuclear magnetic resonance (1H-NMR), Fourier transform infrared (FTIR) spectrometry, and high-performance size-exclusion chromatography (HPSEC) were employed. The results revealed the conjugation of gallic acid with the 15.5 kDa dextran from Leuconostoc mesenteroides. Dex&ndash, Gal had a molecular weight of 11.2 kDa, indicating that the conjugation reaction was accompanied by a minor degradation of Dex&ndash, Gal. In addition, Dex&ndash, Gal contained 36.8 &plusmn, 1.4 mg gallic acid per gram dextran. These molecules were also evaluated as antioxidants using total antioxidant capacity (TAC), reducing power, ferric chelation, and superoxide radical-scavenging assays. Both polysaccharides had no ferric chelation activity. In addition, Dex&ndash, Gal was more efficient as an antioxidant agent in TAC (13 times) and was more efficient than dextran in superoxide radical-scavenging (60 times) and reducing power (90 times) assays. These data demonstrate that Dex&ndash, Gal is a natural-compound-based antioxidant with potential applications in the pharmaceutical, cosmetic, and food industries.

Artemisia annua is cultivated mainly for isolation of artemisinin, a potent antimalarial compound. Moderate salt stress has been proved to increase the artemisinin synthesis by the plant. The aim of this study was to evaluate the influence of salt stress on physiological parameters and cell wall polysaccharides of A. annua. Plants subjected to salt stress displayed reduction in the biomass and length and showed high damage of cellular membranes. Cell wall polysaccharides extracted from aerial parts with hot water, EDTA and NaOH also exhibited modifications in the yield and monosaccharide composition. The main changes were found in the pectic polysaccharides: increase of homogalacturonan domain, increase of neutral side chains and increase in the methyl esterification. 1H NMR analyses of pectins indicated that for A. annua, arabinans have an important role in coping with salt stress. Hemicellulose domain was also modified under salt stress, with increased xylose contents. The results indicated adaptations in the cell wall of A. annua under salt stress.

International audience; We have synthesized polylactide (PLA) polymers containing five distinct lengths of fluorinated end-groups (from C 3 F 7 to C 13 F 27). The influence of fluorinated end-group lengths and their dynamics in chlorinated solvents (chloroform and dichloromethane) was investigated as a function of the presence of perfluorohexane (PFH) and related to the morphology of capsules of PFH obtained using these fluorinated polylactides. 19 F spin−spin relaxation time (T 2) measurements revealed a reduced mobility of the fluorinated units with dependency on fluorinated chain length in the order C 3

Xylan is one of the most abundant hemicellulose constituents in the plant kingdom. We extracted xylan from corn cobs (XCC) using ultrasound. The structure of XCC was determined by NMR analysis, which revealed a composition of xylose:glucose:arabinose:galactose:mannose:glucuronic acid in a molar percentage ratio of 48:21:16:10:2.5:2.5. XCC induced the proliferation of murine macrophage cells (RAW 264.7) and inhibited the proliferation of human lung adenocarcinoma epithelial cells (A549) by 20% and human cervical adenocarcinoma cells (HeLa) by 60%. Several cell death-associated morphological changes were observed after the exposure of HeLa cells to XCC for 24 h. In addition, by using fluorescent probes, we observed a larger number of irregular and pyknotic nuclei with condensed chromatin bodies (nuclear condensation). FACS analysis showed an increase in the number of annexin V-positive and propidium iodide (PI)-positive cells (37.0%) in the presence of XCC compared with that of the negative control cells (5.0%). XCC also increased the ratio of Bax:Bcl-2 (3.5:1) and the levels of cytochrome c, caspase 3, and apoptosis-inducing factor (AIF) in treated cells. Therefore, the results demonstrated the antiproliferative potential of XCC, which induced apoptosis in HeLa cells.

A fucogalactan (E) was isolated from aqueous extract of Agaricus bisporus. The monosaccharide composition, methylation, and NMR analyses showed it is constituted by a (1→6)-linked α- d -Galp main-chain, partially methylated at O-3, and partially substituted at O-2 by non-reducing end-units of α- l -Fucp or α- d -Galp. HPSEC analysis showed it had Mw of 1.28 × 104 g mol−1. The polysaccharide was sulfated modifying reaction time, molar ratio of sulfation agent to hydroxyl group on the polysaccharide (ηClSO3H/OH ratio), and ratio of total reaction volume to weight of sample (VT/w ratio; μL mg−1). The degree of substitution (DS) was evaluated for all sulfated derivatives. The sulfated fucogalactan with the highest DS value (2.83) had the best anticoagulant activity on Activated Partial Thromboplastin Time (APTT) and Protrombin Time (PT) assays. This sulfated fucogalactan, named E100, was obtained with the optimal conditions of ηClSO3H/OH ratio of 18, VT/w ratio of 100, in 6 h of reaction. The results showed that E100 produces a linear increment of APTT for concentrations of 15–45 μg mL−1, whereas PT was almost constant between 20 and 400 μg mL−1, suggesting an anticoagulant activity via inhibition of the intrinsic pathway of blood coagulation. NMR and methylation analyses showed that α- d -Galp units of the main chain were greatly sulfated on 2-O-, 3-O-, and 4-O-positions.

Glucosylceramides (GlcCer) are the main neutral glycosphingolipids expressed in fungal cells. In this work, glucosylceramides (GlcCer) were extracted from three strains of Scedosporium (Pseudallescheria) boydii, one strain of Pseudallescheria ellipsoidea and one strain of Pseudallescheria angusta and purified by several chromatographic steps. Using high-performance thin layer chromatography (HPTLC), we found a similarity between GlcCer obtained from all of the analysed strains. A detailed structural analysis of the P. ellipsoidea GlcCer was performed via electrospray ionization mass spectrometry (ESI-MS) and confirmed in 1- and 2-D heteronuclear NMR experiments ((1)H-(13) C HSQC). GlcCer species produced by mycelial forms of these strains displayed the same structure previously demonstrated by our group for P. boydii, Cryptococcus neoformans, Pseudallescheria minustipora, Fusarium solani, and Colletotrichum gloesporioides. A monoclonal antibody (mAb) against GlcCer was used for immunofluorescence experiments. Our results revealed that GlcCer is present on the surface of these fungi, and no difference was observed in the GlcCer structure of the present set of strains in terms of geographic or clinical origin, suggesting a conserved GlcCer structure similar to those previously described for Scedosporium apiospermum, Scedosporium aurantiacum, and P. minutispora. The surface distribution of GlcCer in these fungi is suggestive of the involvement of this molecule in fungal growth.