Non-Cytotoxic Sulfated Heterorhamnan from Gayralia brasiliensis Green Seaweed Reduces Driver Features of Melanoma Metastatic Progression.

Melanoma is a form of skin cancer with high mortality owing to its fast progression and metastatic capacity. The treatments available nowadays are only palliative in advanced stages of the disease. Thus, alternative therapies for cancer treatment are in demand, and molecules from natural sources, such as polysaccharides, could represent new possible therapeutic approaches. Polysaccharides of freshwater and marine algae with biological activities, such as antitumor properties, are greatly reported in the scientific literature. In the present study, a sulfated heterorhamnan obtained from the green seaweed Gayralia brasiliensis (Gb1 fraction) was chemically characterized and its biological activities in the B16-F10 murine melanoma cell line were evaluated. The Gb1 polysaccharidic fraction tested concentrations presented low or absence of cytotoxicity to B16-F10 cells and neither cell proliferation nor cell cycle were altered. Interestingly, Gb1 treatment decreased B16-F10 cells migration and invasion capabilities and CD44 labeling, showing to be a promising compound for further in vitro and in vivo antitumor studies.

The antiviral activity of sulfated polysaccharides against dengue virus is dependent on virus serotype and host cell.

Two homogeneous sulfated polysaccharides obtained from the red seaweeds Gymnogongrus griffithsiae and Cryptonemia crenulata, the kappa/iota/nu carrageenan G3d and the dl-galactan hybrid C2S-3, were assayed for their antiviral properties against the four serotypes of dengue virus (DENV) in different host cell types. Both seaweed derivatives were selective inhibitors of DENV-2 multiplication in Vero cells with inhibitory concentration 50% (IC50) values around 1 microg/ml and selectivity indices > 1000. The compounds had a lower antiviral effect against DENV-3 (IC50 values in the range 13.9-14.2 microg/ml), an even lower effect against DENV-4 (IC50 values in the range 29.3 to > 50 microg/ml) and were totally inactive against DENV-1. With respect to the host cell, the polysulfates were inhibitors of DENV-2 and DENV-3 in the human hepatoma HepG2 and foreskin PH cells, with similar antiviral effectiveness as in Vero cells, but were totally inactive in mosquito C6/36 HT cells. Mechanistic studies demonstrated that G3d and C2S-3 were active DENV-2 inhibitors only when added together with the virus or early after infection, and both initial processes of virus adsorption and internalization are the main targets of these compounds. Therefore, the variations in antiviral activity of the polysaccharides depending on the viral serotype and the host cell may be ascribed to differences in the virus-cell interaction leading to virus entry.

Protective effect of a natural carrageenan on genital herpes simplex virus infection in mice.

In the present study, the protective effect of 1T1, a lambda-carrageenan extracted from the red seaweed Gigartina skottsbergii was evaluated in a murine model of herpes simplex virus type 2 (HSV-2) genital infection. Six to eight-week-old female BALB/c mice were intravaginally inoculated with a lethal dose of HSV-2 (MS strain) and pre- or post-infection treated with different doses of a 10mg/ml solution of 1T1. A single topical administration of 1T1 shortly before infection of BALB/c mice with HSV-2 protected 9 out of 10 mice from HSV-2-induced lesions and mortality, compared with only 10% survival in control mice. In addition, 1T1 produced a total blockade in virus shedding in the vaginal secretions. When 1T1 pre-treatment was reinforced with a second dose 2h after infection, total protection was observed even when the prophylactic administration had taken place at 60min before infection. The irreversible virucidal action of 1T1 against herpes virus seems to be responsible of its protective effect against virus replication and mortality following vaginal HSV-2 infection.

Structural studies on fucoidans from the brown seaweed Sargassum stenophyllum.

The brown seaweed Sargassum stenophyllum biosynthesizes two different sets of fucoidans. One of them is characterized by higher percentages of glucuronic acid and fewer sulfate groups, which are situated on different sugar units. alpha-L-Fucose was the major component but other sugars like beta-D-galactose, beta-D-mannose, alpha-D-glucuronic acid, alpha-D-glucose and beta-D-xylose were also in substantial amounts. Fucoidans from the other set contain small amounts of alpha-D-glucuronic acid and high percentages of sulfate groups, which are concentrated on the fucose residues, with only fucose and galactose as major components. Structural studies of one fucoidan from each set suggest that these products have a general basic structure that has a formal resemblance to that of the fucosylated chondroitin sulfates from the body wall of sea cucumbers, namely, a linear core (formed mainly by (1-->6)-beta-D-galactose and/or (1-->2)-beta-D-mannose units) with branched chains of 'fucans' (formed by (1-->3) and/or (1-->4)-alpha-L-fucose, (1-->4)-alpha-D-glucuronic acid, terminal beta-D-xylose and, sometimes, (1-->4)-alpha-D-glucose). In fucoidans from the second set, the 'core' is reduced to short galactan chains.

Room temperature, low-field 13C-n.m.r. spectra of degraded carrageenans: Part III. Autohydrolysis of a lambda carrageenan and of its alkali-treated derivative.

Autohydrolysis of lambda and theta carrageenans produces degraded products which are useful for 13C-n.m.r. spectroscopy, with low-field equipment and at room temperature. Data obtained from these spectra are doubly valuable as they are used for the structural study of parent carrageenans and to investigate the reactions carried out during autohydrolysis.

Antiherpetic and anticoagulant properties of carrageenans from the red seaweed Gigartina skottsbergii and their cyclized derivatives: correlation between structure and biological activity.

The antiviral activity against herpes simplex virus types 1 and 2 of kappa/l-, partially cyclized mu/v-, and lambda-carrageenans isolated from the red seaweed Gigartina skottsbergii and their cyclized derivatives was analyzed. lambda-Carrageenans and the partially cyclized mu/v-carrageenan were the most potent inhibitors of herpes viruses (including acyclovir-resistant variants and clinical isolates), with IC50 values lower than 1 microgram ml-1 against both serotypes and selectivity indices higher than 10(3). kappa/l-Carrageenans were slightly less effective than the other two types with IC50 values in the range 1.6-4.1 micrograms ml-1. Antiherpetic activity was directly correlated to the amount of alpha-D-galactose 2,6-disulfate residues in the natural carrageenans. The cyclization of the alpha-D-galactose 6-sulfate and 2,6-disulfate units into 3,6-anhydro-alpha-D-galactose and 3,6-anhydro-alpha-D-galactose 2-sulfate residues in these polysaccharides, in general, lowers the antiherpetic activity of the derivatives with respect to the natural carrageenans. Some carrageenans showed a very reduced anticoagulant activity only at concentrations that were considerably higher than the IC50, whereas others were totally devoid of anticoagulant properties. Among natural carrageenans, the mu/v-type IC3 shows the best relationship between antiviral efficacy and lack of anticoagulant action, resulting a very promising compound.

NMR and rheological study of Aloe barbadensis partially acetylated glucomannan.

The structural and rheological properties of the Aloe extract (AE) and the polysaccharidic fraction (PF) obtained from the leaves pulp of Aloe barbadensis Miller were investigated. Structural analyses carried out by composition, methylation analysis and NMR spectroscopy showed that PF is mainly constituted by a partially acetylated 4-linked ß-d-glucomannan. The acetyl groups are located at C-2, C-2 and C-3, C-3 and/or C-6. The acetylation pattern of this type of polysaccharide was for the first time established using bidimensional NMR analyses. AE and PF aqueous solutions at 25°C showed a non-Newtonian behavior (with pseudoplastic characteristics), however PF showed higher apparent viscosity than AE. Dynamic oscillatory analyses showed that both samples, at the same concentration, behaved as a concentrated solution. PF presented higher values of G' compared with those of AE and this behavior could be consequence of its higher content in partially acetylated glucomannan.

Co-culture of microalgae, cyanobacteria, and macromycetes for exopolysaccharides production: process preliminary optimization and partial characterization.

In this study, the biomass and exopolysaccharides (EPS) production in co-cultures of microalgae/cyanobacteria and macromycetes was evaluated as a technology for producing new polysaccharides for medical and/or industrial application. Based on biomass and EPS productivity of monocultures, two algae and two fungi were selected and cultured in different co-culture arrangements. The hydrosoluble EPS fractions from mono- and co-cultures were characterized by ¹³C NMR spectroscopy and gas chromatography coupled to mass spectrometry and compared. It was found that co-cultures resulted in the production of an EPS different from those produced by monocultures, showing fungal predominance with microalgal/cyanobacterial traces. Co-cultures conditions were screened (temperature, agitation speed, fungal and microalgae inoculation rate, initial pH, illumination rate, and glucose concentration) in order to achieve maximum biomass and EPS production, resulting in an increase of 33 and 61% in exopolysaccharides and biomass productions, respectively (patent pending).

Agar from Gracilaria gracilis (Gracilariales, Rhodophyta) of the Patagonic coast of Argentina--content, structure and physical properties.

Milled summer thalli of Gracilaria gracilis from Argentina were sequentially extracted with water at room temperature (RTW1-3), 70 degrees C (W701-3) and 90 degrees C (W901-2). Both W701 and W901 consisted of high molecular weight polysaccharides (ca. 540,000Da), but polydispersity was higher for the major product W701 (yield, 72% of the recovered). Structural analyzes by methylation and (13)C NMR spectroscopy revealed that W701 was mainly agarose. Alkaline treatment, together with structural analyzes, indicated a negligible proportion of precursor l-galactose 6-sulfate residues in this product, while they were clearly detected in the (13)C NMR spectra of RTW2-3. The presence of floridean starch in W901 had an antagonistic effect on its gel strength, which resulted nearly three times lower than that of fraction W701. Ultrastructural observation by transmission electron microscopy showed that, after extraction with hot water, a partial loss of cell wall stratification and disorganization of the cuticle had occurred. Final cellular debris exhibited swelling in the microfibrillar component. After this first thorough study of the chemical composition and physical properties of the products of G. gracilis from Bahía Bustamante we conclude that a good quality agarose is obtained in high yield after extraction with water at 70 degrees C without the requirement of alkaline pretreatment, which usually produces degradation of the polysaccharide.

Production and characterization of the exopolysaccharides produced by Agaricus brasiliensis in submerged fermentation.

The aim of the work was to study the production of the exopolysaccharides by Agaricus brasiliensis and the isolation of exopolysaccharides (EPSs) with biological effects. A brasiliensis LPB03 was cultured in submerged fermentation in a medium containing glucose, yeast extract, hydrolyzed soybean protein, and salts (pH 6.1) at 29 degrees C and 120 rpm for 144 h. The maximum biomass and EPS yield was 7.80 +/- 0.01 and 1,430.70 +/- 26.75 mg/L, respectively. To isolate the produced EPSs, two methods were compared: (1) with alcohol precipitation and (2) treatment with tricloroacetic acid (TCA), followed by alcohol precipitation. The use of TCA facilitated the purification of the EPS, reducing the amount of the contaminant soy proteins. For monosaccharide identification, the EPSs were hydrolyzed, derivatized to alditol acetates, and analyzed by gas chromatography (GC) and GC-mass spectrometry, which showed the presence (in molar percentage) of mannose (58.7), galactose (21.4), and glucose (13.1) as major sugars, with lower amounts of rhamnose (3.9) and xylose (2.8). Scanning electron microscopy was used to observe the morphological structure of the EPS. The experiments in vivo including EPS in the mice diet during 8 weeks indicated the hipocholesteremic and hypoglycemic effects.

Inhibitory effect of sulfated galactans from the marine alga Bostrychia montagnei on herpes simplex virus replication in vitro.

Sulfated polysaccharides exhibit many biological properties such as antiviral and anticoagulant activities. Herein, we report the antiviral activity of sulfated galactans extracted from the red sea-weed Bostrychia montagnei against herpes simplex virus types 1 (strain F and the thymidine kinase-deficient strains Field and B2006) and 2 (strain G). Two crude extracts obtained with cold and hot water as well as some fractions obtained by anion exchange chromatography, inhibited significantly the replication of the different strains of herpesviruses as determined by plaque reduction assays. The inhibitory effect of the compounds studied here took place only when they were added during the adsorption period. They were found to be highly selective antiviral substances, causing no impairment of Vero cell viability. Furthermore, they had no direct inactivating effect on virions by incubation in a virucidal assay. The antiviral activity could be correlated with the molecular weight and sulfate content of the polysaccharides. Although sulfated polysaccharides are generally endowed with anticoagulant properties, the results of the activated partial thromboplastin time and the thrombine time assays indicated that the natural sulfated polysaccharides from Bostrychia montagnei have very low anticoagulant activity, confirming that there is no relation between the antiviral and anticoagulant properties.