Physical, structural and antioxidant properties of brewer's spent grain protein films.

BACKGROUND: The development of brewer's spent grain protein (BSG-PC) films with potential active packaging properties was investigated. Films were prepared by casting protein dispersions at different pH values (2, 8, 11), plasticizers [polyethylene glycol (PEG) or glycerol] and levels (0-0.25 g g-1 ) of PEG. Mechanical, water-barrier and solubility, optical, antioxidant (reducing power, ABTS•+ and lipidic radical scavenging), and antimicrobial properties of films were determined. Also, the structural characteristics of films were evaluated by attenuated total reflectance-Fourier transform infrared spectroscopy. RESULTS: Only films prepared at pH 2 and plasticized by PEG were homogeneous in appearance and could be manipulated; thus, different levels of PEG were studied at this pH. Higher PEG concentrations increased water solubility, water vapor permeability and elongation at break, and decreased tensile strength and elastic modulus. PEG increased α-helix structure only when 0.10 g PEG g-1 BSG-PC was used. This could be related to the better mechanical properties of F0.10 films (higher tensile strength, and elastic modulus) with respect to the other films. Antioxidant activity depended on PEG concentration, whereas no antimicrobial properties against Bacillus cereus, Salmonella newport and Penicillium corylophylum were detected. CONCLUSION: The formulations with 0.10 and 0.15 g PEG g-1 BSG-PC appear to be the most promising, balancing mechanical, water-barrier properties and the antioxidant capacity of these films. Moreover, BSG proteins could be a cheap alternative for the preparation of biodegradable films, which are capable of being used as active food packaging. © 2020 Society of Chemical Industry.

Molecular action mechanism of anti-inflammatory hydrolysates obtained from brewers' spent grain.

BACKGROUND: Brewers' spent grain (BSG) is a relevant, protein-rich by-product of the brewing process. Protein hydrolysates from different sources exert immune-regulatory actions activating toll-like receptors (TLRs), nuclear factor kappa B (NFκB), and mitogen-activated protein kinases (MAPKs). Effects of gastrointestinal digestion have been poorly studied. Here, we studied the immune-regulatory effect of BSG hydrolysates, and their in-vitro-digested products, on rat splenocytes, macrophages, and T lymphocytes RESULTS: In primary cultures of rat spleen cells, BSG hydrolysates induced interleukin 10 and tumor necrosis factor production in basal conditions. Under stimulation with lipopolysaccharide or concanavalin A, hydrolysates further induced interleukin 10 production. Tumor necrosis factor and interferon-γ were inhibited in lipopolysaccharide- and concanavalin-A-stimulated cells respectively. In vitro gastrointestinal digestion attenuated the observed effects. Splenic macrophages and T lymphocytes behaved in a similar fashion. In spleen cells from TLR2-/- and TLR4-/- mice, immune-regulatory effects were greatly reduced or abrogated. The study of signal transduction pathways indicated a major involvement of NFκB, and the contribution of MAPKs p38, c-Jun N-terminal kinase, and extracellular signal-regulated kinases 1 and 2. CONCLUSION: BSG hydrolysates, like those obtained from other food sources, regulate the immune response, involving TLR2 and TLR4 and the activation of NFκB and MAPKs, an effect partly maintained after in vitro gastrointestinal digestion. Our data support the hypothesis of a shared, rather unspecific, mechanism of action of protein hydrolysates. © 2020 Society of Chemical Industry.

Green Alga Ulva spp. Hydrolysates and Their Peptide Fractions Regulate Cytokine Production in Splenic Macrophages and Lymphocytes Involving the TLR4-NFκB/MAPK Pathways.

Hydrolysates of food protein sources have immunomodulatory effects, which are of interest for use as functional foods. In this study, we have characterized the immune regulatory effect on rat splenocytes, macrophages and T lymphocytes of Ulva spp. hydrolysates and their peptide fractions with or without in vitro gastrointestinal digestion and/or ultrafiltration. IL-10 was induced in almost all conditions and cell types obtained from wild type animals. The induction was in general increased by ultrafiltration and in vitro gastrointestinal digestion. TNF was also induced in basal conditions. In turn, TNF and IFN-γ production was attenuated by the hydrolysate products in lipopolysaccharide or concanavalin A immune stimulated cells. Inhibitors for the activation of NFκB, MAPK p38 and JNK inhibited IL-10 induction in rat splenocytes. The response was dramatically attenuated in TLR4-/- cells, and only modestly in TLR2-/- cells. Food peptides from Ulva spp. genus exert anti-inflammatory effects in immune cells mediated by TLR4 and NFκB. Similarity with the immunomodulatory profile of protein hydrolysates from other sources suggests a common mechanism.

Antithrombotic Activity of Brewers' Spent Grain Peptides and their Effects on Blood Coagulation Pathways.

Antithrombotic activity of brewers' spent grain peptides before and after simulated gastrointestinal digestion and their effects on blood coagulation pathways were evaluated. Two hydrolysates were produced using sequential enzymatic systems: alkaline protease + Flavourzyme (AF) and neutral protease + Flavourzyme (PF). Simulation of gastrointestinal digestion of AF and PF hydrolysates was made using porcine pepsin and pancreatin enzymes, obtaining the corresponding digested samples: AFD and PFD, respectively. Peptides were fractionated by ultrafiltration using a 1 kDa cut-off membrane. Hydrolysates had peptides with medium and low molecular weights (2100 and 500 Da, respectively), and Glu, Asp, Leu, Ala, and Phe were the most abundant amino acids. Gastrointestinal digested hydrolysates presented high proportion of small peptides (~500 Da), and higher amount of Val, Tyr, and Phe than hydrolysates. Mass spectrum (HDMS Q-TOF) of AFD-ultrafiltered fraction <1 kDa exhibited peptides from 500 to 1000 Da, which are not present in AF. PFD showed the generation of new peptides from 430 to 1070 Da. All samples showed thrombin inhibitory activity. However, no effect was observed on prothrombin time. Peptides <1 kDa from hydrolysates and digested samples delayed thrombin and thromboplastin time respect to the control (~63%). Also the samples showed thrombin inhibitory activity at common pathway level. Thus, brewers' spent grain peptides exerted their antithrombotic activity by inhibiting the intrinsic and common pathways of blood coagulation. This is the first report to demonstrate that brewers' spent grain peptides are able to delay clotting time after simulated gastrointestinal digestion.

Colonic and systemic effects of extruded whole-grain sorghum consumption in growing Wistar rats.

Colonic effects of extruded whole-grain sorghum diets were evaluated using a model of growing rats. In all, twenty-four male Wistar rats were fed control (C), extruded white sorghum (EWS) or red sorghum (ERS). Consumption of sorghum diets showed satiety properties, with reduction of caecal pH, and lower activity of ß-glucosidase and ß-glucuronidase enzymes. Decreased copper zinc superoxide dismutase and manganese superoxide dismutase and increased catalase and glutathione peroxidase levels were observed in colonic mucosa. The induction of antioxidant enzymes occurred through the activation of the nuclear factor erythroid 2-related factor 2 protein and its subsequent translocation into the nucleus. ERS was able to decrease the proliferation of proximal mucosa of colon, demonstrating a possible effect against colorectal tumourigenesis. EWS increased proliferation and also apoptosis, ensuring the re-establishment of homoeostasis of the colonic mucosa. No antioxidant systemic effect (serum or hepatic level) was observed. It is likely that despite the extrusion the low bioavailability of the phenolic compounds of sorghum diets caused them to exert mainly acute effects at the colon level. Extruded whole-grain sorghum is a good functional ingredient that might be promising in dietary prevention of intestinal diseases.

Bioactive Properties of Phaseolus lunatus (Lima Bean) and Vigna unguiculata (Cowpea) Hydrolyzates Incorporated into Pasta. Residual Activity after Pasta Cooking.

The aims of the study were to study the inclusion of P. lunatus (PLH) and V. unguiculata (VUH) protein hydrolyzates with bioactive properties into a pasta-extruded product and determine residual activity after extrusion or pasta cooking. Both protein hydrolyzates showed angiotensin-converting enzyme inhibition (ACEI) and antioxidant activity (TEAC). PLH showed higher ACEI but lower TEAC than VUH (97.19 ± 0.23 vs. 91.95 ± 0.29 % and 244.7 ± 3.4 vs. 293.7 ± 3.3 µmol Trolox/g, respectively). They were included at 5 or 10 % into wheat pasta. Control pasta had the lowest ACEI activity or TEAC (22.01 ± 0.76 % or 14.14 ± 1.28 µmol Trolox/g, respectively). Higher activity remained in pasta with PLH than VUH after extrusion, and higher the level of addition, higher the ACEI was. Pasta had practically the same ACEI activity after cooking, thus active compounds were not lost by temperature or lixiviation. Regarding TEAC, higher activity remained in pasta with 10 % VUH (31.84 ± 0.17 µmol Trolox/g). Other samples with hydrolyzates had the same activity. After cooking, pasta with hydrolyzates had higher TEAC values than control, but these were not modified by the level of incorporation. Moreover, the profile changed because pasta with PLH had the highest TEAC values (21.39 ± 0.01 and 20.34 ± 0.15 for 5 or 10 % hydrolyzates, respectively). Cooking decreased this activity (~ 20 %), for all samples. Although a certain loss of antioxidant activity was observed, pasta could be a good vehicle for bioactive compounds becoming a functional food.

Chelating Properties of Peptides from Red Seaweed Pyropia columbina and Its Effect on Iron Bio-Accessibility.

The aim of this work was to evaluate copper-chelating, iron-chelating and anticariogenic activity of peptides obtained by enzymatic hydrolysis of P. columbina protein concentrate and to study the effects of chelating peptides on iron bio-accessibility. Two hydrolyzates were obtained from P. columbina protein concentrate (PC) using two hydrolysis systems: alkaline protease (A) and alkaline protease + Flavourzyme (AF). FPLC gel filtration profile of PC shows a peak having molecular weight (MW) higher than 7000 Da (proteins). A and AF hydrolyzates had peptides with medium and low MW (1013 and 270 Da), respectively. Additionally, AF presented free amino acids with MW around 82 Da and higher content of His and Ser. Peptides from AF showed the highest chelating properties measured as copper-chelating activity (the lowest ß-carotene oxidation rate: Ro; 0.7 min(-1)), iron-chelating activity (33%), and phosphorous and Ca(2+) release inhibition (87 and 81%, respectively). These properties could indicate antioxidant properties, promotion of iron absorption and anticariogenic activity, respectively. In fact, hydrolyzates promoted iron dialyzability (≈ 16%), values being higher than that found for P. columbina seaweed. Chelating peptides from both hydrolyzates can maintain the iron in a soluble and bio-accessible form after gastrointestinal digestion.

Proteins and Carbohydrates from Red Seaweeds: Evidence for Beneficial Effects on Gut Function and Microbiota.

Based on their composition, marine algae, and namely red seaweeds, are good potential functional foods. Intestinal mucosal barrier function refers to the capacity of the intestine to provide adequate containment of luminal microorganisms and molecules. Here, we will first outline the component of seaweeds and will summarize the effects of these on the regulation of mucosal barrier function. Special attention will be paid to unique components of red seaweeds: proteins and derived peptides (e.g., phycobiliproteins, glycoproteins that contain "cellulose binding domains", phycolectins and the related mycosporine-like amino acids) together with polysaccharides (e.g., floridean starch and sulfated galactans, such as carrageenans, agarans and "dl-hybrid") and minerals. These compounds have been shown to exert prebiotic effects, to regulate intestinal epithelial cell, macrophage and lymphocyte proliferation and differentiation and to modulate the immune response. Molecular mechanisms of action of peptides and polysaccharides are starting to be elucidated, and evidence indicating the involvement of epidermal growth factor receptor (EGFR), insulin-like growth factor receptor (IGFR), Toll-like receptors (TLR) and signal transduction pathways mediated by protein kinase B (PKB or AKT), nuclear factor-κB (NF-κB) and mitogen activated protein kinases (MAPK) will also be summarized. The need for further research is clear, but in vivo experiments point to an overall antiinflammatory effect of these algae, indicating that they can reinforce membrane barrier function.

Chemical composition, nutritional and antioxidant properties of the red edible seaweed Porphyra columbina.

Proximate composition, fatty acids and amino acid profiles and nutritional (chemical score, protein digestibility, PDCAAS and mineral dialyzability) and antioxidant properties (TEAC, DPPH and power reduction) from Porphyra columbina were evaluated. Total dietary fiber (48.02 ± 1.13 g/100 g dry weight) and protein (24.61 ± 0.21 g/100 g dry weight) were the two most abundant components in this seaweed. The main saturated and unsaturated fatty acids were C16:0 and C20:5 (n-3), respectively. The limiting amino acid was tryptophan with a chemical score of 57%. Protein digestibility was 74.33 ± 3.0%. Porphyra columbina has high mineral content with good Na/K relationship and medium value of potential mineral accessibility (P, Ca and Zn dializability: 18.75 ± 0.01, 17.62 ± 0.16 and 16.70 ± 0.44, respectively). The highest antioxidant properties were obtained with an acetone/water extraction system. This work provides important information about chemical composition and nutraceutical new properties of P. columbina.

Anti-diabetic properties of brewer's spent yeast peptides. In vitro, in silico and ex vivo study after simulated gastrointestinal digestion.

Brewer's spent yeast (BSY) hydrolysates are a source of antidiabetic peptides. Nevertheless, the impact of in vitro gastrointestinal digestion of BSY derived peptides on diabetes has not been assessed. In this study, two BSY hydrolysates were obtained (H1 and H2) using ß-glucanase and alkaline protease, with either 1 h or 2 h hydrolysis time for H1 and H2, respectively. These hydrolysates were then subjected to simulated gastrointestinal digestion (SGID), obtaining dialysates D1 and D2, respectively. BSY hydrolysates inhibited the activity of α-glucosidase and dipeptidyl peptidase IV (DPP-IV) enzymes. Moreover, although D2 was inactive against these enzymes, D1 IC50 value was lower than those found for the hydrolysates. Interestingly, after electrophoretic separation, D1 mannose-linked peptides showed the highest α-glucosidase inhibitory activity, while non-glycosylated peptides had the highest DPP-IV inhibitory activity. Kinetic analyses showed a non-competitive mechanism in both cases. After peptide identification, GILFVGSGVSGGEEGAR and IINEPTAAAIAYGLDK showed the highest in silico anti-diabetic activities among mannose-linked and non-glycosylated peptides, respectively (AntiDMPpred score: 0.70 and 0.77). Molecular docking also indicated that these peptides act as non-competitive inhibitors. Finally, an ex vivo model of mouse jejunum organoids was used to study the effect of D1 on the expression of intestinal epithelial genes related to diabetes. The reduction of the expression of genes that codify lactase, sucrase-isomaltase and glucose transporter 2 was observed, as well as an increase in the expression of Gip (glucose-dependent insulinotropic peptide) and Glp1 (glucagon-like peptide 1). This is the first report to evaluate the anti-diabetic effect of BSY peptides in mouse jejunum organoids.

In vivo and in silico study of antioxidant and anti-inflammatory effects on the liver-spleen axis of microencapsulated brewers' spent grain peptides.

Metabolic syndrome (MS) is a cluster of risk factors for the development of cardiovascular disease and type 2 diabetes mellitus. Some dietary bioactive compounds such as peptides can exert dual antioxidant and anti-inflammatory effects. The aim of this study was to analyze the effects of microencapsulated brewers' spent grain peptides (BSG-P-MC) on hepatic injury, lipid peroxidation, oxidative stress and inflammation in the liver-spleen axis in Wistar rats fed with a sucrose-rich diet (SRD). Male rats received for 100 days a reference diet (RD), SRD or RD and SRD containing 700 mg per kg body weight per day of BSG-P-MC. The results demonstrated that BSG-P-MC reversed injury, lipid peroxidation, and oxidative stress in the liver. For the spleen, BSG-P-MC decreased the levels of lipid peroxidation, CAT activity, NFκB, PAI-1 and F4/80 protein mass levels with respect to the SRD-fed rats. Three peptides identified by LC-MS/MS from BSG-P-MC after in vitro gastrointestinal digestion showed high in silico free radical scavenging activity (LPRDPYVDPMAPLPR, ANLPRDPYVDPMAPLPRSGPE and ANLPRDPYVDPMAPLPR). Moreover, two identified peptides presented high in silico anti-inflammatory properties (LTIGDTVPNLELDSTHGKIR and VDPDEKDAQGQLPSRT). This study is the first report of antioxidant and anti-inflammatory properties of microencapsulated BSG-peptides exerted in the liver-spleen axis in a MS rodent model.

Microencapsulated bioactive peptides from brewer's spent grain promotes antihypertensive and antidiabetogenic effects on a hypertensive and insulin-resistant rat model.

The effects of microcapsules containing brewer's spent grain (BSG) peptides were evaluated on a hypertensive/insulin-resistant rat model induced by a sucrose-rich diet (SRD) administration. Animals received for 100 days the control diet (CD), SRD, and CD and SRD diets supplemented with microencapsulated peptides (CD-P and SRD-P). During the experimental period, blood pressure was monitored. Glycemia, tissue glycogen content, nitric oxide, and the activity of enzymes related to hypertensive and diabetogenic mechanisms were determined. The consumption of SRD caused hypertensive and hyperglycemic effects compared to CD. However, the SRD-P group presented lower systolic pressure at the middle of ingestion, achieving similar values than the CD. The SRD-P rats decreased all enzymes' activities compared to the SRD reaching the values of CD, except for those of α-amylase in cecal content and DPP-IV in serum. It was possible to corroborate potential antihypertensive and antidiabetogenic in vivo effects of the microencapsulated BSG peptides. PRACTICAL APPLICATIONS: Brewer's spent grain (BSG) is the main waste obtained from brewing industry. Bioactive peptides obtained after an enzymatic hydrolysis of proteins with in vitro antihypertensive and antidiabetogenic activity have been described. However, to corroborate the action of these bioactive peptides, in vivo studies are necessary. In the present work, microcapsules containing bioactive peptides from BSG were administered on the rat model with induced hypertension and insulin-resistance, corroborating an in vivo antihypertensive and antidiabetogenic effects by inhibition of enzymes related with blood pressure regulation and glucose metabolism. This work demonstrated that microcapsules of BSG peptides could be included into functional foods formulations, or used as dietary supplement for improving health and the prevention of non-communicable diseases, adding value to the brewing process by-product.

Effect of extrusion process on antioxidant and ACE inhibition properties from bovine haemoglobin concentrate hydrolysates incorporated into expanded maize products.

Extrusion process has been widely used for the development of many functional foods. The aim of this study was to assess the effect of extrusion process on antioxidant and angiotensin-converting enzyme (ACE) inhibition properties from bovine haemoglobin concentrate (BHC) hydrolysates (P, FC, PF and FCF). Extrusion was carried out with a Brabender single screw extruder. The ACE inhibition and the antioxidant capacity (AC) were estimated by the inhibition of the ACE and ABTS+√ radical cation expressed as Trolox equivalent antioxidant capacity (TEAC), respectively. The ACE inhibition and TEAC values from hydrolysates were significantly higher than that from BHC. The highest ACE inhibition corresponded to P hydrolysate and the highest TEAC corresponded to PF and FCF hydrolysates. The ACE inhibition and AC from extruded products with added hydrolysates were higher than that from maize control; however, the extrusion process modified both ACE inhibition and AC formerly present in hydrolysates.

Iron and zinc availability and some physical characteristics from extruded products with added concentrate and hydrolysates from bovine hemoglobin.

Four hydrolysates were obtained from bovine hemoglobin concentrate (BHC) and used to fortify extruded maize products. Extrusion was carried out with a Brabender single-screw extruder. Physicochemical properties from extruded products were measured. The iron availability was estimated by the dializability method, which measures the mineral dialyzed after a double digestion simulating physiological processes. The physicochemical properties of the extruded products were not affected by fortification, with the exception of total soluble solids. The enzymic hydrolysis increased the iron dializability with respect to the substrate. The highest value of iron dializability corresponded to the more hydrolysated sample. Extruded products fortified with BHC hydrolysates showed higher iron dializability than those fortified with BHC. However, iron dializability corresponding to BHC is lower than that expected from heme iron. Therefore, heme-iron availability is low when it is determined in the absence of meat proteins, and hydrolysis could increase potential iron availability.

Development of naturally activated edible films with antioxidant properties prepared from red seaweed Porphyra columbina biopolymers.

The aim of this work was to study the physicochemical and antioxidant properties of phycobiliproteins-phycocolloids-based films, obtained from mixtures of two aqueous fractions extracted from Porphyra columbina red seaweed, one enriched in phycocolloids (PcF) and the other in phycobiliproteins (PF). Films with different ratios of PF:PcF (0, 25, 50, 75, 100% [w/w]) and without plasticizer addition were prepared by casting. PcF films had excellent mechanical properties (tensile strength ∼50MPa, elongation at break ∼3% and an elastic modulus ∼17.5MPa). The addition of PF to formulations exerted a plasticizing effect on the PcF matrix, which was manifested in moisture content, water solubility and mechanical properties of the resulting films but not in its water vapour permeability. The antioxidant capacity (TEAC) of the PcF films was significantly increased by the addition of PF and a direct relationship between TEAC and the total phenolic compounds (r(2)=0.9998) and R-phycoerythrin (r(2)=0.9942) was observed.

Composición química y calidad proteica de fideos complementados con harina de Porphyra columbina / Chemical composition and protein quality of pasta complemented with Porphyra columbina

La harina de trigo es un alimento no balanceado, debido al bajo valor biológico de su proteína, originado por la deficiencia de lisina. Para mejorar las propiedades nutricionales de la pasta de trigo, se utilizan distintas fuentes con alto contenido y calidad de proteína. Las algas son un recurso abundante, económico y atractivo para utilizar como ingrediente en alimentos. Entre las macroalgas rojas comestibles, Porphyra columbina es una de las más importantes de la Patagonia Argentina. El objetivo de este trabajo fue comparar la composición química y la calidad proteica de fideos elaborados con harina de Triticum durum (FC) y fideos complementados al 30% en la fase sólida con harina de Porphyra columbina (F30). La composición centesimal se determinó por métodos oficiales de la AOAC. La determinación cuantitativa de aminoácidos se llevó a cabo mediante el método propuesto por Alaiz et al. (23). La separación de los aminoácidos se realizó mediante HPLC. Los resultados de composición química para FC y F30, expresados en g cada 100 g de pasta seca comestible, fueron respectivamente: humedad 7,9 ± 0,4 y 8,1 ± 0,2; cenizas 1,0 ± 0,1 y 1,7 ± 0,1; grasas 5,6 ± 0,2 y 4,7 ± 0,1; proteínas 15,8 ± 0,1 y 17,7 ± 0,1; fibra dietaria total 7,7 ± 0,3 y 19,8 ± 0,2 y carbohidratos 62,0 y 47,9. Los cálculos del puntaje químico, teniendo en cuenta las necesidades del preescolar, revelaron un valor de 55% y 94% para FC y F30, respectivamente. En FC la lisina se evidenció como el aminoácido limitante, mientras que en F30 el limitante fue el triptofano(AU).

The wheat pasta is an unbalanced food, due to the low biological value of its protein, caused by the deficiency of lysine. To improve the nutritional properties of wheat pasta, different sources with high protein content and quality are used. Seaweed is an abundant, economical and attractive resource to use as an ingredient in food. Among red edible macroalgae, Porphyra columbina is one of the most important ones in Argentine Patagonia. The objective of this work was to compare the chemical composition and protein quality of pasta made with Triticum durum flour (FC) and pasta supplemented at 30% in the solid phase with Porphyra columbina flour (F30). The centesimal composition was determined by AOAC official methods. The quantitative determination of amino acids was carried out by the method proposed by Alaiz, et al.(23). The separation of amino acids was carried out through HPLC. The chemical composition results for FC and F30, expressed in g per 100 g of edible dry pasta, were: water 7,9 ± 0,4 and 8,1 ± 0,2; ash 1,0 ± 0,1 and 1,7 ± 0,1; fat 5,6 ± 0,2 and 4,7 ± 0,1; protein 15,8 ± 0,1 and 17,7 ± 0,1; total dietary fiber 7,7 ± 0,3 and 19,8 ± 0,2 and carbohydrates 62,0 and 47,9, respectively. The chemical score, taking into account the needs of pre-schoolers, revealed a value of 55% and 94% for FC and F30, respectively. In FC, lysine was found to be the limiting amino acid, whereas in F30, the limiting factor was tryptophan(AU).

A Porphyra columbina hydrolysate upregulates IL-10 production in rat macrophages and lymphocytes through an NF-κB, and p38 and JNK dependent mechanism.

The marine environment represents a relatively untapped source of functional ingredients. Here we characterise a hydrolysate obtained from Phorphyra columbina (PcRH) and its effects on primary splenocytes, macrophages and T lymphocytes in vitro. Our product had a high degree of hydrolysis, due to the use of a mixture of endo-peptidase and exo-peptidase, and was enriched in Asp, Ala and Glu. PcRH had mitogenic effects on rat splenic lymphocytes. IL-10 secretion was enhanced by PcRH in splenocytes (235%), macrophages (150%) and in lymphocytes (472%), while the production of TNFα and other proinflammatory cytokines by macrophages was inhibited (15-75%), especially under lipopolysaccharide stimulation. The effect of the hydrolysate on IL-10 was evoked by JNK, p38 MAPK and NF-κB dependent pathways in T lymphocytes. We conclude that PcRH has immunomodulatory effects on macrophages and lymphocytes, activating NF-κB and MAPK dependent pathways, and predominantly inducing IL-10 production.

Immunomodulatory properties of the protein fraction from Phorphyra columbina.

The phycobiliproteins from Rhodophyta , R-phycoerythrin (R-PE) and C-phycocyanin (C-PC), have been shown to exert immunomodulatory effects. This study evaluated the effects of a Phorphyra columbina protein fraction (PF) and R-PE and C-PC on rat primary splenocytes, macrophages, and T-lymphocytes in vitro. PF featured various protein species, including R-PE and C-PC. PF showed mitogenic effects on rat splenocytes and was nontoxic to cells except at 1 g L(-1) protein. IL-10 secretion was enhanced by PF in rat splenocytes, macrophages, and especially T-lymphocytes, whereas it was markedly diminished by R-PE and C-PC. The production of pro-inflammatory cytokines by macrophages was inhibited. The effect of PF on IL-10 was evoked by JNK/p38 MAPK and NF-κB-dependent pathways in macrophages and T-lymphocytes. It was concluded that PF has immunomodulatory effects on macrophages and lymphocytes that appear to be predominantly anti-inflammatory via up-regulated IL-10 production and cannot be accounted for by R-PE and C-PC.