In vitro anti-tumoral and immunomodulatory effects of acidic polysaccharides isolated from the fungus Gloeosoma mirabile.

Polysaccharides from wood-rooting fungi have attracted attention due to their broad pharmacological properties. Herein, we report the antitumor and immunomodulatory activities of acid polysaccharides isolated from fungi Gloeosoma mirabile. The polysaccharide extracts displayed significant antiproliferative activity against cancer cell lines (MCF-7, HCT-116, U-937) in a dose-dependent manner and induction of IL-6 in macrophage RAW 264.7. Furthermore, flow cytometry analysis showed that high polysaccharide concentrations induced apoptosis by 83% in HL-60 cells. Based on gas chromatography-mass spectrometry (GC-MS) and Fourier transform infra-red (FT-IR) spectroscopy studies, acidic polysaccharides from G. mirabile were mainly composed of arabinose, α-D-galactopyranose and methyl ß-D-galactopyranoside.

Combined Dietary Administration of Chlorella fusca and Ethanol-Inactivated Vibrio proteolyticus Modulates Intestinal Microbiota and Gene Expression in Chelon labrosus.

The use of functional feeds in aquaculture is currently increasing. This study aimed to assess the combined impact of dietary green microalgae Chlorella fusca and ethanol-inactivated Vibrio proteolyticus DCF12.2 (CVP diet) on thick-lipped grey mullet (Chelon labrosus) juvenile fish. The effects on intestinal microbiota and the transcription of genes related to metabolism, stress, and the immune system were investigated after 90 days of feeding. Additionally, the fish were challenged with Aeromonas hydrophila and polyinosinic-polycytidylic acid (poly I:C) to evaluate the immune response. Microbiota analysis revealed no significant differences in alpha and beta diversity between the anterior and posterior intestinal sections of fish fed the control (CT) and CVP diets. The dominant genera varied between the groups; Pseudomonas and Brevinema were most abundant in the CVP group, whereas Brevinema, Cetobacterium, and Pseudomonas were predominant in the CT group. However, microbial functionality remained unaltered. Gene expression analysis indicated notable changes in hif3α, mhcII, abcb1, mx, and tnfα genes in different fish organs on the CVP diet. In the head kidney, gene expression variations were observed following challenges with A. hydrophila or poly I:C, with higher peak values seen in fish injected with poly I:C. Moreover, c3 mRNA levels were significantly up-regulated in the CVP group 72 h post-A. hydrophila challenge. To conclude, incorporating C. fusca with V. proteolyticus in C. labrosus diet affected the microbial species composition in the intestine while preserving its functionality. In terms of gene expression, the combined diet effectively regulated the transcription of stress and immune-related genes, suggesting potential enhancement of fish resistance against stress and infections.

In Vitro and In Vivo Effects of Ulvan Polysaccharides from Ulva rigida.

One of the main bioactive compounds of interest from the Ulva species is the sulfated polysaccharide ulvan, which has recently attracted attention for its anticancer properties. This study investigated the cytotoxic activity of ulvan polysaccharides obtained from Ulva rigida in the following scenarios: (i) in vitro against healthy and carcinogenic cell lines (1064sk (human fibroblasts), HACAT (immortalized human keratinocytes), U-937 (a human leukemia cell line), G-361 (a human malignant melanoma), and HCT-116 (a colon cancer cell line)) and (ii) in vivo against zebrafish embryos. Ulvan exhibited cytotoxic effects on the three human cancer cell lines tested. However, only HCT-116 demonstrated sufficient sensitivity to this ulvan to make it relevant as a potential anticancer treatment, presenting an LC50 of 0.1 mg mL-1. The in vivo assay on the zebrafish embryos showed a linear relationship between the polysaccharide concentration and growth retardation at 7.8 hpf mL mg-1, with an LC50 of about 5.2 mg mL-1 at 48 hpf. At concentrations near the LC50, toxic effects, such as pericardial edema or chorion lysis, could be found in the experimental larvae. Our in vitro study supports the potential use of polysaccharides extracted from U. rigida as candidates for treating human colon cancer. However, the in vivo assay on zebrafish indicated that the potential use of ulvan as a promising, safe compound should be limited to specific concentrations below 0.001 mg mL-1 since it revealed side effects on the embryonic growth rate and osmolar balance.

In vitro cytotoxic capacity against tumor cell lines and antioxidant activity of acidic polysaccharides isolated from the Andean Patagonian fungus Phylloporia boldo.

Fungal polysaccharides possess a broad biological activity, including cytotoxic and antioxidant activities. This work aimed to evaluate the cytotoxic and antioxidant activity of the acidic polysaccharides of Phylloporia boldo strain (named PBAP40). Cytotoxic activity of polysaccharide was evaluated determining the viability of three tumor cell lines by MTT assay. The effect of acidic polysaccharide on the cell cycle of HL-60 cell line was evaluated by flow cytometry, and the antioxidant activity was determined by DPPH and ABTS assays. PBAP40 showed cytotoxic effects in tumor cell lines. Results suggest that P. boldo acidic polysaccharides arrested tumor cells in the cell cycle Sub G1 phase. The acidic polysaccharides of PBAP40 strain were not cytotoxic for the non-tumor cell line. PBAP40 also showed excellent antioxidant activity. The FT-IR analysis of the acidic polysaccharides indicated the presence of glucans bearing α- and ß- type glycosidic bonds.

Antioxidant, Immunomodulatory and Potential Anticancer Capacity of Polysaccharides (Glucans) from Euglena gracilis G.A. Klebs

The present study was carried out to determine the bioactivity of polysaccharides extracted from Euglena gracilis (EgPs). These were characterized by FT-IR and GC-MS. Cytotoxicity analyses (MTT) were performed on healthy human gingival fibroblast cell lines (HGF-1), obtaining an IC50 of 228.66 µg mL-1, and cell lines with anticancer activity for colon cancer (HCT-116), breast cancer (MCF-7), human leukemia (U-937, HL-60) and lung cancer (NCl-H460), showing that EgPs have anticancer activity, mainly in HTC-116 cells (IC50 = 26.1 µg mL-1). The immunological assay determined the immunomodulatory capacity of polysaccharides for the production of proinflammatory cytokines IL-6 and TNF-α in murine macrophages (RAW 264.7) and TNF-α in human monocytes (THP-1). It was observed that the EgPs had a stimulating capacity in the synthesis of these interleukins. The antioxidant capacity of polysaccharides and their biomass were analyzed using the ABTS method (18.30 ± 0.14% and (5.40 ± 0.56%, respectively, and the DPPH method for biomass (17.79 ± 0.57%). We quantitatively profiled HGF-1 proteins by liquid chromatography-tandem mass spectrometry analysis, coupled with 2-plex tandem mass tag labelling, in normal cells. In total, 1346 proteins were identified and quantified with high confidence, of which five were considered to be overexpressed. The data is available through ProteomeXchange, under identifier PXD029076.

Sulfated Polysaccharide Extracted from the Green Algae Codium bernabei: Physicochemical Characterization and Antioxidant, Anticoagulant and Antitumor Activity.

Codium bernabei is a green alga that grows on Chilean coasts. The composition of its structural polysaccharides is still unknown. Hence, the aim of this work is to isolate and characterize the hot water extracted polysaccharide fractions. For this purpose, the water extracts were further precipitated in alcohol (TPs) and acid media (APs), respectively. Both fractions were characterized using different physicochemical techniques such as GC-MS, GPC, FTIR, TGA, and SEM. It is confirmed that the extracted fractions are mainly made of sulfated galactan unit, with a degree of sulfation of 19.3% (TPs) and 17.4% (ATs) and a protein content of 3.5% in APs and 15.6% in TPs. Other neutral sugars such as xylose, glucose, galactose, fucose, mannose, and arabinose were found in a molar ratio (0.05:0.6:1.0:0.02:0.14:0.11) for TPs and (0.05:0.31:1.0:0.03:0.1:0.13) for ATs. The molecular weight of the polysaccharide samples was lower than 20 kDa. Both polysaccharides were thermally stable (Tonset > 190 °C) and showed antioxidant activity according to the ABTS•+ and DPPH tests, where TPs fractions had higher scavenging activity (35%) compared to the APs fractions. The PT and APTTS assays were used to measure the anticoagulant activity of the polysaccharide fractions. In general, the PT activity of the TPs and APs was not different from normal plasma values. The exception was the TPs treatment at 1000 µg mL−1 concentration. The APTTS test revealed that clotting time for both polysaccharides was prolonged regarding normal values at 1000 µg mL−1. Finally, the antitumor test in colorectal carcinoma (HTC-116) cell line, breast cancer (MCF-7) and human leukemia (HL-60) cell lines showed the cytotoxic effect of TPs and APs. Those results suggest the potential biotechnological application of sulfate galactan polysaccharides isolated from a Chilean marine resource.

Immunomodulatory, Antioxidant Activity and Cytotoxic Effect of Sulfated Polysaccharides from Porphyridium cruentum. (S.F.Gray) Nägeli.

Porphyridium cruentum is a unicellular microalga that can synthesize and secrete to the culture medium-high amounts of polysaccharides. In this study, the immunomodulatory, cytotoxic effect and antioxidant activity of the sulfated polysaccharides (PcSPs) were determinate. The PcSPs were precipitated with 2% Cetylpyridinium bromide hydrate and ethanol and purified by dialysis. The extract was lyophilized for its characterization by Fourier transform-Infrared (FT-IR) spectroscopy and gas chromatography-mass spectrometry (GC-MS). The antioxidant activity of PcSPs were examined with assay 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) and compared with that of the biomass, observing significant differences between the results obtained from the PcSPs and biomass. To determine their ability to induce cytokine production Tumor Necrosis Factor alpha (TNF-α) and interleukina-6 (IL-6), the immunomodulatory activity of the PcSPs has been evaluated. In the mouse macrophage cell line (RAW 264.7), PcSPs are potent inducers of IL-6 cytokines but mainly of TNF-α. The cytotoxic capacity of PcSPs was measured by the MTT colorimetric assay in colorectal carcinoma (HTC-116), human leukemia (U-937 and HL-60), breast cancer (MCF-7), lung cancer (NCI-H460) and human gingival fibroblasts (HGF-1) cell lines. The IC50 value of 2311.20 µg mL-1, 1676.74 µg mL-1, 1089.63 µg mL-1, 5498.14 µg mL-1 and 2861.49 µg mL-1 respectively in the tumor lines and 5022.55 µg mL-1 in gingival fibroblasts were obtained. Our study suggested that PcSPs from P. cruentum have a moderate immunomodulatory and cytotoxic effect. The results obtained indicate that the polysaccharides from P. cruentum are potent inducers of IL-6 cytokines and, most importantly, of TNF-α. PcSPs showed no evidence of antigenic activity or hypersensitivity when administered intraperitoneally in mice. Furthermore, the in vivo study revealed an improvement of local inflammatory response against stress in the peritoneum. These findings suggest that the PcSPs from P. cruentum might have potential as a valuable ingredient in nutraceutical products.

Immunomodulatory and Antioxidant Activities of Sulfated Polysaccharides from Laminaria ochroleuca, Porphyra umbilicalis, and Gelidium corneum.

Seaweeds of the genera Laminaria, Gelidium, and Porphyra have been used in both food and non-food industries due to their unique properties and characteristic biological activity. This study assesses the antioxidant activity and immunomodulatory properties of the acidic polysaccharides extracted from Laminaria ochroleuca, Porphyra umbilicalis, and Gelidium corneum collected in the Atlantic coast of Tarifa (Cadiz, Spain). The proliferation of murine cell line RAW 264 decreased with increasing concentration of polysaccharides of the three algal species. The highest both antioxidant (25.69 µmol TE g-1 DW) and immunomodulatory activities were observed in the sulfated polysaccharides of L. ochroleuca compared to that of P. umbilicalis and G. corneum. Sulfated polysaccharides of L. ochroleuca presented high potential anticancer activity in cell lines of human colon cancer HTC-116 (IC50 = 0.44 mg mL-1), human malignant melanoma G-361 (IC50 = 5.42 mg mL-1), breast adenocarcinoma human MCF-7 (IC50 = 8.32 mg mL-1), and human leukemia U-937 (IC50 = 3.72 mg mL-1). It is concluded that metabolites of L. ochroleuca can offer significant advantages for the pharmaceutical industry, particularly when macrophage activation is required.

UV Photoprotection, Cytotoxicity and Immunology Capacity of Red Algae Extracts.

This study was designed to evaluate the potential use of algal extracts in cosmeceuticals, including factors related to biosecurity. The aqueous crude extracts of Hydropuntia cornea and Gracilariopsis longissima showed a good photoprotective capacity (Sun Protection Factor, SPF) due to, among other reasons, the presence of five types of mycosporine-like amino acids (MAAs) detected by high pressure liquid chromatography-photodiode array detector (HPLC-PDA) and electrospray ionization mass spectrometry (ESI-MS) (Palythine, Asterina-330, Shinorine, Porphyra-334, and Palythinol). The toxicity of the extracts was evaluated by the MTT assay, which is based on the metabolic reduction of MTT [3-(4,5-dimethylthiazol-2yl)-diphenyl tetrazolium bromide] by the action of the mitochondrial enzyme succinate dehydrogenase. This assay was carried out in vitro in three cell lines: one related to the immune system (murine macrophages of the immune system: RAW264.7) and two human cell lines related to the skin (gingival fibroblasts: HGF, and immortalized human keratinocytes: HaCaT). Both extracts showed no cytotoxic activity in both types of human cells, whereas they showed cytotoxicity in murine tumor cells of the immune system (macrophages: RAW264.7). On the other hand, the immunological activity in the murine macrophage RAW264.7 was studied at a concentration lower than 100 µg mL-1 and lower than the EC50, and evaluated by the production of pro-inflammatory compounds through an immunosorbent assay linked to enzymes such as tumor necrosis factor-α (TNF-α) or anti-inflammatory/proinflammatory enzymes such as interleukin-6 (IL-6). Both algae extracts induced the biosynthesis of TNF-α and IL-6. The production of TNF-α was much higher than that observed in the control (at a concentration of the aqueous extract higher than 5 µg mL-1). These results support the theory that the extracts of H. cornea and G. longissima actively induce the production of cytokines. In summary, the extracts of these species did not show cytotoxicity in human cells, and they present with immunomodulatory and photoprotection capacity.

Resource-allocation tradeoffs in caddisflies facing multiple stressors.

The replacement of native forests by exotic tree monocultures, such as those of Eucalyptus, decreases the quality of leaf litter inputs to streams and often reduces riparian cover, which can elevate water temperature. The combined effects of these stressors on the survival and performance of detritivores may be important, as detritivore species loss leads to reduced litter breakdown, a key ecosystem process. Potential loss of cased caddisfly larvae is of particular concern because they are the predominant detritivores in many streams, they are sensitive to warming, and they expend energy on building and carrying their cases, which may be an added burden under times of stress. In a microcosm experiment, we tested whether (i) poor-quality Eucalyptus globulus litter impaired case construction by larvae of Sericostoma pyrenaicum (due to preferential allocation of the scarcer available energy to larval fitness) compared to high-quality Alnus glutinosa litter; (ii) whether this effect was enhanced by higher temperatures (15 vs. 10°C) resulting in faster metabolism and greater energy expenditure; but (iii) reduced in the presence of chemical cues from a predatory fish (due to greater investment in more protective cases). We found that Eucalyptus had lethal and sublethal effects on larval caddisflies, increasing mortality, reducing growth, and impairing case construction, compared to larvae fed Alnus. Temperature did not reinforce the effects of exotic litter on case construction, but predator chemical cues triggered the construction of more protective cases (i.e., longer and better cemented) despite the lower resource quality, providing evidence for environmentally mediated resource-allocation tradeoffs.

Combinaciones enzimáticas en la obtención de hidrolizados proteicos a partir de Chlorella vulgaris / Enzimatic combinations in obtaining protein hydrolisates from Chlorella vulgaris

Las microalgas representan en la actualidad una opción atractiva para el desarrollo de nuevos suplementos nutricionales y alimentos para regímenes especiales, mediante la bioconversión de la energía solar en virtud del alto contenido de proteínas y sustancias biológicamente activas en la biomasa celular. Se investigó el efecto de la utilización de 5 combinaciones enzimáticas en la hidrólisis de las proteínas presentes en la biomasa autotrófica de la microalga Chlorella vulgaris tratada previamente con etanol. Los mejores resultados se obtuvieron con las mezclas papaína-tripsina y papaína-pancreatina, las que rindieron hidrolizados con niveles de nitrógeno amínico y grados de hidrólisis superiores al 4 y 45 por ciento, respectivamente. Estos elementos evidencian un efecto positivo del empleo de mezclas enzimáticas en comparación con las enzimas individuales, lo cual está mediado por la superposición de sus acciones hidrolíticas(AU)

Combinaciones enzimáticas en la obtención de hidrolizados proteicos a partir de Chlorella vulgaris / Enzimatic combinations in obtaining protein hydrolisates from Chlorella vulgaris

Las microalgas representan en la actualidad una opción atractiva para el desarrollo de nuevos suplementos nutricionales y alimentos para regímenes especiales, mediante la bioconversión de la energía solar en virtud del alto contenido de proteínas y sustancias biológicamente activas en la biomasa celular. Se investigó el efecto de la utilización de 5 combinaciones enzimáticas en la hidrólisis de las proteínas presentes en la biomasa autotrófica de la microalga Chlorella vulgaris tratada previamente con etanol. Los mejores resultados se obtuvieron con las mezclas papaína-tripsina y papaína-pancreatina, las que rindieron hidrolizados con niveles de nitrógeno amínico y grados de hidrólisis superiores al 4 y 45 por ciento, respectivamente. Estos elementos evidencian un efecto positivo del empleo de mezclas enzimáticas en comparación con las enzimas individuales, lo cual está mediado por la superposición de sus acciones hidrolíticas

Adyuvantes inmunológicos / Immunoadjuvants

La búsqueda de nuevas sustancias con actividad adyuvante/inmunopotenciadora constituye una de las tendencias más importantes en la investigación inmunológica actual. El presente trabajo considera los criterios fundamentales que determinan la selección de un adyuvante con fines investigativos, en el campo de la salud veterinaria y en la formulación de vacunas humanas, con el objetivo de incrementar la efectividad de la respuesta inmune. Se analizan brevemente las principales dificultades identificadas en la literatura, relacionadas con la comparación de la efectividad de diferentes adyuvantes, y se destacan las amplias posibilidades que ofrecen los productos naturales, especialmente polisacáridos de diversas fuentes, en la tamización de sustancias con propiedades inmunoestimulantes(AU)

Adyuvantes inmunológicos / Immunoadjuvants

La búsqueda de nuevas sustancias con actividad adyuvante/inmunopotenciadora constituye una de las tendencias más importantes en la investigación inmunológica actual. El presente trabajo considera los criterios fundamentales que determinan la selección de un adyuvante con fines investigativos, en el campo de la salud veterinaria y en la formulación de vacunas humanas, con el objetivo de incrementar la efectividad de la respuesta inmune. Se analizan brevemente las principales dificultades identificadas en la literatura, relacionadas con la comparación de la efectividad de diferentes adyuvantes, y se destacan las amplias posibilidades que ofrecen los productos naturales, especialmente polisacáridos de diversas fuentes, en la tamización de sustancias con propiedades inmunoestimulantes