Chondroitin Sulfate from Oreochromis niloticus Waste Reduces Leukocyte Influx in an Acute Peritonitis Model.

Oreochromis niloticus (tilapia) is one of the most cultivated fish species worldwide. Tilapia farming generates organic waste from fish removal processes in nurseries. Visceral waste can damage natural ecosystems. Therefore, the use of this material as a source of biomolecules helps reduce environmental impacts and improve pharmacological studies. Tilapia viscera were subjected to proteolysis and complexation with an ion-exchange resin. The obtained glycosaminoglycans were purified using ion exchange chromatography (DEAE-Sephacel). The electrophoretic profile and analysis of 1H/13C nuclear magnetic resonance (NMR) spectra allowed for the characterization of the compound as chondroitin sulfate and its sulfation position. This chondroitin was named CST. We tested the ability of CST to reduce leukocyte influx in acute peritonitis models induced by sodium thioglycolate and found a significant reduction in leukocyte migration to the peritoneal cavity, similar to the polymorphonuclear population of the three tested doses of CST. This study shows, for the first time, the potential of CST obtained from O. niloticus waste as an anti-inflammatory drug, thereby contributing to the expansion of the study of molecules with pharmacological functions.

In Vitro Antitumor Potential of Sulfated Polysaccharides from Seaweed Caulerpa cupressoides var. flabellata.

Seaweeds are important source of bioactive compounds, including sulfated polysaccharides (SP). Because of their structural heterogeneity, these compounds are promising sources of anticancer compounds. SP from brown and red seaweeds have shown antimelanoma activity in different in vitro and in vivo models. However, SP from green seaweed are still poorly evaluated. Therefore, SP were extracted from the green alga Caulerpa cupressoides var. flabellata, and their antiproliferative, anti-migratory, and inhibitory effect on melanin production on B16-F10 melanoma cells was evaluated. Cell assays, including flow cytometry, demonstrated that SP (100-1000 µg mL-1) are non-cytotoxic, do not induce apoptosis or necrosis, and do not interfere with cell cycle. However, SP (1000 µg mL-1) were found to significantly inhibit cell colony formation (80-90%), cell migration (40-75%), and melanin production (~ 20%). In summary, these results showed that SP inhibited important melanoma development events without cytotoxicity effects, suggesting that C. cupressoides may be an important source of SP with antitumor properties.

Structural characterization of blackberry wine polysaccharides and immunomodulatory effects on LPS-activated RAW 264.7 macrophages.

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-Manp(1 → 6)-linked units, O-2 substituted with α-d-Manp(1 → 2)-linked side chains for BWPFp and a AG II formed by a major chain of ß-d-Galp(1 → 3)-linked, substituted at O-6 by side chains of the ß-d-Galp(1 → 6)-linked, which then are substituted at O-3 by non-reducing units of α-l-Araf and a RG I, formed by [→4)-α-d-GalpA-(1 → 2)-α-l-Rhap-(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-1ß) in LPS-treated cells.

Structural characterization of polysaccharides from Cabernet Franc, Cabernet Sauvignon and Sauvignon Blanc wines: Anti-inflammatory activity in LPS stimulated RAW 264.7 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 ß-d-Galp main chain, substituted at O-6 by (1 → 6)-linked ß-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-1ß) and mediator (NO) in RAW 264.7 cells stimulated with LPS.

Heparin fails to inhibit the leukocyte recruitment for an extended time following inflammatory stimulus.

CONTEXT: Several studies have shown that heparin is able to inhibit leukocyte recruitment during an early acute inflammatory response. However, considering the pharmacokinetic aspects of heparin and the dynamics of inflammation our objective was to determine if heparin is able to retain its antimigratory property during a prolonged inflammatory response. OBJECTIVE: Compare the effect of heparin on leukocyte recruitment to the peritoneal cavity during early acute inflammatory response and for a longer time post-inflammatory stimulus. MATERIALS AND METHODS: Wistar rats pre-treated with subcutaneous heparin in doses of 1, 5, and 15 µg/kg were challenged with 2 mL intraperitoneal thioglycollate. After 3 or 8 h, the animals were killed. The cells in the peritoneal cavity were collected and counted. For differential counting, cells from peritoneal lavage and from blood were distended over a glass slide, stained, and counted. RESULTS: After 3 h, heparin inhibited cell influx to the injury site at all tested dosages. The largest effect was achieved at a 5 µg/kg dose (83% of reduction, p < 0.001). After 8 h, heparin at a 1 µg/kg dose reduced 63% of cellular infiltration (p < 0.001); the group treated with a 15 µg/kg dose presented an pro-inflammatory effect observed by the higher proportions, when compared with the thioglycollate group, of neutrophils on whole blood (60.9%, p < 0.001) and peritoneal fluid (27.3%, p < 0.05), and whole blood monocytes (117.8%, p < 0.01). CONCLUSION: These findings show that the heparin effect on leukocyte recruitment varies depending on its dosage and the duration of the inflammation.