Multiple heteroatom dopant carbon dots as a novel photoluminescent probe for the sensitive detection of Cu2+ and Fe3+ ions in living cells and environmental sample analysis.

Heavy metal ion pollution harms human health and the environment and continues to worsen. Here, we report the synthesis of boron (B), phosphorous (P), nitrogen (N), and sulfur (S) co-doped carbon dots (BP/NS-CDs) by a one-step facile hydrothermal process. The optimum synthetic parameters are of 180 °C temperature, 12 h reaction time and 15% of PBA mass. The as-synthesized BP/NS-CDs exhibits excellent water solubility, strong green photoluminescence (PL) at 510 nm, and a high quantum yield of 22.4%. Moreover, BP/NS-CDs presented high monodispersity (7.2 ± 0.45 nm), excitation-dependent emission, PL stability over large pH, and high ionic strength. FTIR, XRD, and XPS are used to confirm the successful B and P doping of BP/NS-CDs. BP/NS-CD photoluminescent probes are selectively quenched by Cu2+ and Fe3+ ions but showed no response to the presence of other metal cations. The PL emission of BP/NS-CDs exhibited a good linear correlation with Cu2+ and Fe3+ concentrations with detection limits of 0.18 µM and 0.27 µM for Cu2+ and Fe3+, respectively. Furthermore, the HCT116 survival cells kept at 99.4 ± 1.3% and cell imaging capability, when the BP/NS-CDs concentration is up to 300 µg/mL by MTT assay. The proposed sensor is potential applications for the detection of Cu2+ and Fe3+ ions in environmental water samples.

Nitrogen and sulfur co-doped photoluminescent carbon dots for highly selective and sensitive detection of Ag+ and Hg2+ ions in aqueous media: Applications in bioimaging and real sample analysis.

In this study, we report the synthesis of photoluminescent (PL) nitrogen (N) and sulfur (S) co-doped carbon dots (NS-CDs) from nitazoxanide and 3-mercaptopropionic acid as a precursors via a one-pot hydrothermal methods. N and S co-doped materials allows more active sites in the CDs surface resulting in an enhancement of their PL properties. NS-CDs show bright blue PL, excellent optical properties, good water solubility, and a high quantum yield (QY) of 32.1%. The as-prepared NS-CDs were confirmed by UV-Visible, photoluminescence, FTIR, XRD and TEM analysis. An optimized excitation at 345 nm, the NS-CDs exhibited strong PL emission at 423 nm with an average size of 3.53 ± 0.25 nm. Under optimized conditions, the NS-CDs PL probe shows high selectivity with Ag+/Hg2+ ions detected, while other cations no significant changes the PL signal. The PL intensity of NS-CDs linearly quenching and enhancement with Ag+ and Hg2+ ions from 0 to 50 × 10-6 M, with the detection limit of 2.15 × 10-6 M and 6.77 × 10-7 M (S/N = 3). More interestingly, as-synthesized NS-CDs shows a strong binding to Ag+/Hg2+ ions with the PL quenching and enhancement to precise and quantitative detection of Ag+/Hg2+ ions in living cells. The proposed system was effectively utilized for the sensing of Ag+/Hg2+ ions in real samples resulting in high sensitivity and good recoveries (98.4-109.7%).

In Vitro Anti-Inflammatory Activity and Structural Characteristics of Polysaccharides Extracted from Lobonema smithii Jellyfish.

Crude polysaccharides were extracted from the white jellyfish (Lobonema smithii) using water extraction and fractionated using ion-exchange chromatography to obtain three different fractions (JF1, JF2, and JF3). The chemical characteristics of four polysaccharides were investigated, along with their anti-inflammatory effect in LPS-stimulated RAW264.7 cells. All samples mainly consisted of neutral sugars with minor contents of proteins and sulphates in various proportions. Glucose, galactose, and mannose were the main constituents of the monosaccharides. The molecular weights of the crude polysaccharides and the JF1, JF2, and JF3 fractions were 865.0, 477.6, 524.1, and 293.0 kDa, respectively. All polysaccharides were able to decrease NO production, especially JF3, which showed inhibitory activity. JF3 effectively suppressed iNOS, COX-2, IL-1ß, IL-6, and TNF-α expression, while IL-10 expression was induced. JF3 could inhibit phosphorylated ERK, JNK, p38, and NF-κB p65. Furthermore, flow cytometry showed the impact of JF3 on inhibiting CD11b and CD40 expression. These results suggest that JF3 could inhibit NF-κB and MAPK-related inflammatory pathways. The structural characterisation revealed that (1→3)-linked glucopyranosyl, (1→3,6)-linked galactopyranosyl, and (1→3,6)-linked glucopyranosyl residues comprised the main backbone of JF3. Therefore, L. smithii polysaccharides exhibit good anti-inflammatory activity and could thus be applied as an alternative therapeutic agent against inflammation.

An effective bio-inspired synthesis of platinum nanoparticles using Caulerpa sertularioides and investigating their antibacterial and antioxidant activities.

In this study, we report the synthesis of platinum nanoparticles (Cs-PtNPs) using an aqueous extract of Caulerpa sertularioides as a reducing agent. Cs-PtNPs were characterized by UV-Vis spectroscopy, fourier transform infrared (FT-IR) spectroscopy, X-ray diffraction (XRD), field emission electron microscopy (FE-SEM), energy-dispersive X-ray spectroscopy (EDAX), high-resolution transmission electron microscopy (HR-TEM) and dynamic light scattering (DLS) analysis. Cs-PtNPs are spherical with a particle size of 6-22 nm. Cs-PtNPs have been shown to have highly effective antioxidant activities with 74% for DPPH, 63% for reducing power, and 59% for total antioxidant at 1 mg/ml, and results were compared with standard L-ascorbic acid. Furthermore, the Cs-PtNPs demonstrated excellent antibacterial activity against the Gram-negative bacteria, Vibrio parahaemolyticus with the highest zone of inhibition (18 mm) at 50 µg/ml. Moreover, Artemia nauplii showed less toxicity when treated with Cs-PtNPs at 150 µg/ml, indicating that the Cs-PtNPs are less toxic and environment friendly.

Effects of Dietary Fish Meal Replacement with Alternative Protein Ingredients and Their Combinations on Growth Performance, Feed Utilization, Fillet Composition, and Biochemical Parameters of Red Seabream (Pagrus major).

The experiment was conducted to evaluate alternative protein ingredients in a low-fish meal (FM) diet for red seabream (Pagrus major). Twelve experimental diets were formulated. Control diet (CON) was designed to contain 60% FM. Other experimental diets were formulated by replacing 50% of FM from the CON with soy protein concentrate (SPC), corn gluten (CG), meat meal (MM), and/or chicken byproduct meal (CBM). Four diets were designed including one of SPC, CG, MM, or CBM as FM replacer and designated as SPC, CG, MM, and CBM. Six other diets were formulated by adding two ingredients as SPC and CG, SPC and MM, SPC and CBM, CG and MM, CG and CBM, or MM and CBM, and designated as SCG, SMM, SCM, CMM, CCM, and MCM, respectively. The 12th diet (MIX) was formulated by including SPC, CGM, MM, and CBM. Triplicate fish groups (50.2 ± 0.1 g) were hand-fed for 12 weeks. Weight gain (WG) of fish was significantly improved by MM and MCM diets compared to CG, SCG, CMM, and CCM diets. WG of CON, SPC, CM, SMM, SCM, and MIX groups were comparable with MM and MCM groups. The lowest WG was observed in CG and CMM groups. Feed efficiency (FE) was significantly higher in MM group compared to SPC, CG, SGC, and CMC groups. FE of MCM group was significantly higher than CG and SCG groups. Fillet linolenic acid (C18:2n-6) level in CG group was significantly higher than CON, MM, CM, SCM, CCM, and MCM groups. Serum lysozyme activity was significantly higher in MCM and MIX groups. Therefore, a high level of dietary CG reduces the growth performance and feed utilization of red seabream. A mixture of MM and CBM seems to be more efficient in replacing FM from red seabream diet.

Surveillance of disease incidence in shrimp farms located in the east coastal region of India and in vitro antibacterial efficacy of probiotics against Vibrio parahaemolyticus.

We surveyed 130 shrimp farms located on the eastern coast of India to determine the prevalence of emerging diseases in Litopenaeus vannamei and Penaeus monodon. Live shrimps were collected from the farms based on external symptoms. The biochemical, molecular, and histopathology results confirmed infection with Enterocytozoon hepatopenaei (32.4%), Vibrio parahaemolyticus (27.7%), White Spot Syndrome Virus (25.4%), Vibrio alginolyticus (16.1%), Vibrio harveyi (13.1%), Monodon-type baculovirus (4.61%), and infectious Hematopoietic Necrosis Virus (2.3%) in the collected shrimps. Enterocytozoon hepatopenaei (EHP) occurred more frequently in L. vannamei than P. monodon, with the microsporidian spores in the hepatopancreas. In P. monodon, Monodon-type Baculovirus infection (33.3%) was dominant and small percentages of WSSV, IHHNV, V. alginolyticus, and V. harveyi were observed. A few ponds were observed with co-infection of EHP and WSSV (7.6%), V. parahaemolyticus and WSSV (4.6%) and also V. parahaemolyticus and EHP (6.1%). Among the Vibrio spp, V. parahaemolyticus showed the highest percentage of infection in L. vannamei. Overall, we found that shrimp were chiefly infected with EHP and V. parahaemolyticus. The impact of water quality parameters on shrimp diseases was not addressed in this study. In an antibiotic susceptibility study, V. parahaemolyticus isolated from L. vannamei ponds was susceptible to nitrofurantoin, chloramphenicol, oxytetracycline and tetracycline, but resistant to erythromycin and nalidixic acid. In a preliminary in vitro antibacterial activity assay, probiotics against V. parahaemolyticus showed high inhibitory activity and the results encourage further in-depth studies on the efficacy of probiotics for disease control and prevention in shrimp farms.

Intranasal Administration of Codium fragile Polysaccharide Elicits Anti-Cancer Immunity against Lewis Lung Carcinoma.

Natural polysaccharides have shown promising effects on the regulation of immunity in animals. In this study, we examined the immune stimulatory effect of intranasally administered Codium fragile polysaccharides (CFPs) in mice. Intranasal administration of CFPs in C57BL/6 mice induced the upregulation of surface activation marker expression in macrophages and dendritic cells (DCs) in the mediastinal lymph node (mLN) and the production of interleukin-6 (IL-6), IL-12p70, and tumor necrosis factor-α in bronchoalveolar lavage fluid. Moreover, the number of conventional DCs (cDCs) was increased in the mLNs by the upregulation of C-C motif chemokine receptor 7 expression, and subsets of cDCs were also activated following the intranasal administration of CFP. In addition, the intranasal administration of CFPs promoted the activation of natural killer (NK) and T cells in the mLNs, which produce pro-inflammatory cytokines and cytotoxic mediators. Finally, daily administration of CFPs inhibited the infiltration of Lewis lung carcinoma cells into the lungs, and the preventive effect of CFPs on tumor growth required NK and CD8 T cells. Furthermore, CFPs combined with anti-programmed cell death-ligand 1 (PD-L1) antibody (Ab) improved the therapeutic effect of anti-PD-L1 Ab against lung cancer. Therefore, these data demonstrated that the intranasal administration of CFP induced mucosal immunity against lung cancer.

Neutral Lipids, Glycolipids, and Phospholipids, Isolated from Sandfish (Arctoscopus japonicus) Eggs, Exhibit Anti-Inflammatory Activity in LPS-Stimulated RAW264.7 Cells through NF-κB and MAPKs Pathways.

Total lipids were extracted from sandfish (Arctoscopus japonicus), and then they were separated into the following three lipid fractions: neutral lipids, glycolipids, and phospholipids. In this study, we analyzed the lipid fractions of A. japonicus eggs and we determined their anti-inflammatory activity in RAW264.7 macrophage cells. In these three lipid-fractions, the main fatty acids were as follows: palmitic acid (16:0), oleic acid (18:1n-9), docosahexaenoic acid (DHA, 22:6n-3), and eicosapentaenoic acid (EPA, 20:5n-3). Among the lipid fractions, phospholipids showed the highest concentration of DHA and EPA (21.70 ± 1.92 and 18.96 ± 1.27, respectively). The three lipid fractions of A. japonicus significantly suppressed the production of NO in macrophages. Moreover, they also significantly inhibited the expression of iNOS, COX-2, IL-6, IL-1ß, and TNF-α, in a dose-dependent manner. Furthermore, the lipid fractions of A. japonicus suppressed the nuclear translocation of NF-κB p65 subunits in a dose-dependent manner. In addition, they attenuated the activation of MAPKs (p38, ERK1/2, and JNK) phosphorylation in LPS-stimulated RAW264.7 cells. These results indicate that all the lipid fractions of A. japonicus exert anti-inflammatory activity by suppressing the activation of NF-κB and MAPK pathways. Therefore, the lipid fractions of A. japonicus might be potentially used as anti-inflammatory agents.

Human Peripheral Blood Dendritic Cell and T Cell Activation by Codium fragile Polysaccharide.

Natural polysaccharides exhibit an immunostimulatory effect with low toxicity in humans and animals. It has shown that polysaccharide extracted from Codium fragile (CFP) induces anti-cancer immunity by dendritic cell (DC) activation, while the effect of CFP has not examined in the human immune cells. In this study, we found that CFP promoted the upregulation of CD80, CD83 and CD86 and major histocompatibility complex (MHC) class I and II in human monocyte-derived dendritic cells (MDDCs). In addition, CFP induced the production of proinflammatory cytokines in MDDCs. Moreover, CFP directly induced the activation of Blood Dendritic Cell Antigen (BDCA)1+ and BDCA3+ subsets of human peripheral blood DCs (PBDCs). The CFP-stimulated BDCA1+ PBDCs further promoted activation and proliferation of syngeneic CD4 T cells. The CFP-activated BDCA3+ PBDCs activated syngeneic CD8 T cells, which produced cytotoxic mediators, namely, cytotoxic T lymphocytes. These results suggest that CFP may be a candidate molecule for enhancing immune activation in humans.

Polysaccharide from Codium fragile Induces Anti-Cancer Immunity by Activating Natural Killer Cells.

Natural polysaccharides exhibit beneficial immune modulatory effects, including immune stimulatory and anti-cancer activities. In this study, we examined the effect of Codium fragile polysaccharide (CFP) on natural killer (NK) cell activation, and its effect on tumor-bearing mice. Intravenous CFP treatment of C57BL/6 mice resulted in the upregulation of CD69, which is a marker associated with NK cell activation. In addition, intracellular levels of interferon (IFN)-γ and the cytotoxic mediators perforin and granzyme B were markedly increased in response to the CFP treatment of splenic NK cells. IFN-γ production by NK cells was directly induced by CFP, whereas the upregulation of CD69 and cytotoxic mediators required IL-12. Finally, intraperitoneal treatment with CFP prevented CT-26 (murine carcinoma) tumor cell infiltration in the lungs, without significantly reducing the body weight. In addition, treatment with CFP prevented B16 melanoma cell infiltration in the lung of C57BL/6 mice. Moreover, the anti-tumor effect was diminished by the depletion of NK cells. Therefore, these data suggest that CFP may be used as an NK cell stimulator to produce a phenomenon that contributes to anti-cancer immunity.

Bioengineered gold nanoparticles from marine seaweed Acanthophora spicifera for pharmaceutical uses: antioxidant, antibacterial, and anticancer activities.

In this study, we report a green synthesis of pharmaceutically active gold nanoparticles from marine red alga Acanthophora spicifera by the reduction of chloroauric acid. The formation of A. spicifera-mediated gold nanoparticles (As-AuNPs) was characterized by several analytical techniques. The crystalline and face-centered cubic (fcc) structure were confirmed by X-ray diffraction (XRD) analysis. Electron microscopy results confirmed that As-AuNPs were spherical and the average size of particles was < 20 nm. As-AuNPs hold a significant level of antioxidant activities than A. spicifera extract. As-AuNPs exhibited the highest antibacterial activity against Vibrio harveyi than Staphylococcus aureus at 100 µg/ml. Furthermore, As-AuNPs exhibited the utmost cytotoxicity against human colon adenocarcinoma (HT-29) cells and registered the half-maximal inhibitory concentration (IC50) at 21.86 µg/ml. These findings authenticated that the synthesized As-AuNPs possess a broad spectrum of biological activities, and it can be effectively applied in the field of aquaculture and biomedical application.

Codium fragile F2 sensitize colorectal cancer cells to TRAIL-induced apoptosis via c-FLIP ubiquitination.

This study demonstrates that combined treatment with subtoxic doses of Codium extracts (CE), a flavonoid found in many fruits and vegetables, and tumor necrosis factor-related apoptosis-inducing ligand (TRAIL), induces apoptosis in TRAIL-resistant colorectal cancer (CRC) cells. Effective induction of apoptosis by combined treatment with CE and TRAIL was not blocked by Bcl-xL overexpression, which is known to confer resistance to various chemotherapeutic agents. While TRAIL-mediated proteolytic processing of procaspase-3 was partially blocked in various CRC cells treated with TRAIL alone, co-treatment with CE efficiently recovered TRAIL-induced caspase activation. We observed that CE treatment of CRC cells did not change the expression of anti-apoptotic proteins and pro-apoptotic proteins, including death receptors (DR4 and DR5). However, CE treatment markedly reduced the protein level of the short form of the cellular FLICE-inhibitory protein (c-FLIPS), an inhibitor of caspase-8, via proteasome-mediated degradation. Collectively, these observations show that CE recovers TRAIL sensitivity in various CRC cells via down-regulation of c-FLIPS.

Anti-Inflammatory Effects of Lipids Extracted from Arctoscopus japonicus Eggs on LPS-Stimulated RAW264.7 Cells.

Arctoscopus japonicus is a cold-water marine fish. The present study investigated the fatty acid composition of A. japonicus egg lipids and their anti-inflammatory effects on LPS-stimulated RAW246.7 macrophages. The results showed that A. japonicus egg lipids contained primarily polyunsaturated fatty acids (52.9% of the total fatty acid content; mostly eicosapentaenoic acid [EPA, 21.2 ± 0.5%] and docosahexaenoic acid [DHA, 25.9 ± 0.1%]), followed by monounsaturated fatty acids and saturated fatty acids (23.7% and 23.4%, respectively). A. japonicus egg lipids significantly decreased nitric oxide (NO) production and suppressed the expression of immune-associated genes such as iNOS, COX-2, IL-1ß, IL-6, and TNF-α LPS-stimulated RAW246.7 macrophages in dose-dependent manner. A. japonicus egg lipids also reduced the phosphorylation levels of NF-κB p-65, p38, ERK1/2, and JNK, key components of the NF-κB and MAPK pathways, suggesting that the lipid-induced anti-inflammatory activity is related to these signaling pathways. These results indicate that the lipids extracted from A. japonicus eggs have potential biofunctions and might be useful for regulating inflammation in macrophages.

Bioactivities of Nizamuddinia zanardinii sulfated polysaccharides extracted by enzyme, ultrasound and enzyme-ultrasound methods.

Sulfated polysaccharide (fucoidan) was isolated from Nizamuddinia zanardinii by enzyme (alcalase), ultrasonic and enzyme-ultrasonic methods. The extracted fucoidans were assessed for their chemical compositions, molecular characteristics, anticancer and immunomodulatory activities. Enzyme-ultrasonic isolated fucoidan showed the maximum extraction yield (7.87%) while that obtained by ultrasonic had the minimum value (3.6%). fucoidans were composed of different levels of carbohydrates (52.78-58.65%), proteins (6.98-8.91%), sulfates (21.78-29.6%) and uronic acids (0.42-1.08%). The weight mean average molecular weight of fucoidans varied between 443.7 and 1020.85 kDa. The polysaccharide chains were consisted of fucose, galactose, glucose, mannose and xylose. All the recovered fucoidans showed strong growth inhibition against HeLa and Hep-G2 cancer cells. The isolated fucoidans were non-toxic and considerably stimulated the macrophage cells to release nitric oxide. Enzyme extraction produced fucoidan with the most macrophage stimulation capacity (> 42 µmol). These results suggested that enzyme treatment preserved more sulfate groups in fucoidan structure influencing its anticancer and immunostimulatory activities.

Structural characterization and RAW264.7 murine macrophage stimulating activity of a fucogalactoglucan from Colpomenia peregrina.

Water-soluble polysaccharides were isolated from Colpomenia peregrina to determine their chemical characteristics and immunomodulatory properties. High extraction yields were obtained for CP1 (17.6%) and CP2 (5.2%) polysaccharides. Polysaccharides were mainly consisted of neutral sugars (67.01-73.79%), uronic acids (9.43-14.89%), proteins (3.44-14.89%) and small amounts of sulfates (4.87-4.91%). Polysaccharides were composed of fucose (20.62-24.56%), galactose (25.5-26.94%) and glucose (50.00-52.91%) residues. The average molecular weights of the CP1 and CP2 polysaccharides were 1890 × 103 g/mol and 639 × 103 g/mol, respectively. The polysaccharides exerted a relatively low cytotoxicity against HeLa cancer cells (< 40%). The CP1 and CP2 polysaccharides were nontoxic and induced RAW264.7 murine macrophage cells to release considerable amounts of nitric oxide (NO). Inflammatory cytokines including IL-1ß, TNF-α, IL-6, IL-10 and IL-12 from were secreted from RAW264.7 cells induced with CP1 polysaccharides. As the most immunostimulating fraction, CP1 polysaccharides were homogeneous and formed of 1,3-linked galactose, 1,4-linked glucose and 1,3-linked fucose residues. Overall, these findings suggested that the polysaccharides isolated from C. peregrina can be utilized as potential natural immunostimulant in functional foods or pharmaceutical industries.

Improved immunomodulatory and antioxidant properties of unrefined fucoidans from Sargassum angustifolium by hydrolysis.

The effect of polymer degradation was studied on immunomodulatory and antioxidant properties of fucoidan isolated from S. angustifolium. Partially hydrolyzed fucoidans were prepared using 0.01 N hydrochloric acid after incubation for 10 and 15 min in boiling water. FT-IR analysis showed two major peaks at 850 cm-1 corresponding to bending vibration of C-O-S of sulfate and 1256 cm-1 derived from the stretching vibration of S-O. The native fucoidan consisted mainly of carbohydrate (49.4%), sulfate (22.9%), uronic acid (10.3%) and minor amount of protein (4.1%). The hydrolysis reduced the molecular weight of native fucoidan from 421 × 103 g/mol to 104.1 × 103 g/mol after 10 min boiling and 63.9 × 103 g/mol after 15 min boiling, without a significant change in their chemical compositions. Acid degradation increased the specific volume of gyration from 0.84 to 3.32 cm3/g in hydrolyzed fucoidan polymers. Fucoidan with the lowest molecular weight showed the greatest proliferating effect on RAW264.7 cells and induced the macrophage cells to release more nitric oxide (39.0 µmol) at 50 µg/mL. The DPPH radical scavenging activity, ABTS radical scavenging activity and reducing power remarkably increased after hydrolysis. The current results showed that molecular weight has determinant effect on immunomodulatory and antioxidant activities of unrefined fucoidan and thus acid hydrolysis can be applied on commercial scale to obtain fucoidans with more beneficial effects.

Structural characterization of sulfated arabinans extracted from Cladophora glomerata Kützing and their macrophage activation.

Water-soluble sulfated heteropolysaccharides were extracted from Cladophora glomerata Kützing and fractionated by ion-exchange chromatography, which yielded two subfractions, F1 and F2. The crude and fractionated polysaccharides (F1 and F2) mostly consisted of carbohydrates (62.8-74.5%) with various amounts of proteins (9.00-17.3%) and sulfates (16.5-23.5%), including different levels of arabinose (41.7-54.4%), galactose (13.5-39.0%), glucose (0.80-10.6%), xylose (6.84-13.4%), and rhamnose (0.20-2.83%). Based on the size exclusion chromatography (SEC) profiles, the crude and fractions mainly contained one peak with shoulders having molecular weight (Mw) ranges of 358-1,501 × 10(3). The F1 fraction stimulated RAW264.7 cells to produce considerable amounts of nitric oxide and cytokines compared to the crude and F2 fraction. The backbone of the most potent immunostimulating fraction (F1) was α-(1→4)-L-arabinopyranoside with galactose and xylose residues as branches at O-2 position, and sulfates mainly at O-2 position as well.

Characterization and immunomodulatory activities of polysaccharides from Spirogyra neglecta (Hassall) Kützing.

Sulfated polysaccharides (SP) isolated from freshwater green algae, Spirogyra neglecta (Hassall) Kützing, and fractionated SPs were examined to investigate their molecular characteristics and immunomodulatory activity. The crude and fractionated SPs (F1, F2, and F3) consisted mostly of carbohydrates (68.5-85.3%), uronic acids (3.2-4.9%), and sulfates (2.2-12.2%) with various amounts of proteins (2.6-17.1%). D-galactose (23.5-27.3%), D-glucose (11.5-24.8%), L-fucose (19.0-26.7%), and L-rhamnose (16.4-18.3%) were the major monosaccharide units of these SPs with different levels of L-arabinose (3.0-9.4%), D-xylose (4.6-9.8%), and D-mannose (0.4-2.3%). The SPs contained two sub-fractions with molecular weights (Mw) ranging from 164 × 10(3) to 1460 × 10(3) g/mol. The crude and fractionated SPs strongly stimulated murine macrophages, producing considerable amounts of nitric oxide and various cytokines via up-regulation of their mRNA expression by activation of nuclear factor-kappa B and mitogen-activated protein kinases pathways. The main backbone of the most immunoenhancing SP was (1→3)-L-Fucopyranoside, (1→4,6)-D-Glucopyranoside, and (1→4)-D-Galactopyranoside.

Fermentation enrichment, structural characterization and immunostimulatory effects of ß-glucan from Quinoa.

We developed an efficient mixed-strain co-fermentation method to increase the yield of quinoa ß-glucan (Q+). Using a 1:1 mass ratio of highly active dry yeast and Streptococcus thermophilus, solid-to-liquid ratio of 1:12 (g/mL), inoculum size of 3.8 % (mass fraction), fermentation at 32 °C for 27 h, we achieved the highest ß-glucan yield of (11.13 ± 0.80)%, representing remarkable 100.18 % increase in yield compared to quinoa ß-glucan(Q-) extracted using hot water. The structure of Q+ and Q- were confirmed through Fourier Transform Infrared (FTIR) and Nuclear Magnetic Resonance (NMR) spectroscopies. Q+ contained 41.66 % ß-glucan, 3.93 % protein, 2.12 % uronic acid; Q- contained 37.21 % ß-glucan, 11.49 % protein, and 1.73 % uronic acid. The average molecular weight of Q+(75.37 kDa) was lower than that of Q- (94.47 kDa). Both Q+ and Q- promote RAW264.7 cell proliferation without displaying toxicity. They stimulate RAW264.7 cells through the NF-κB and MAPK signaling pathways, primarily inducing NO and pro-inflammatory cytokines by upregulating CD40 expression. Notably, Q+ exhibited stronger immunostimulatory activity compared to Q-. In summary, the fermentation enrichment method yields higher content of quinoa ß-glucan with increased purity and stronger immunostimulatory properties. Further study of its bioimmunological activity and structure-activity relationship may contribute to the development of new immunostimulants.

Preparation of novel sulfated polysaccharide-carboxymethyl-5-fluorouracil-folic acid conjugates for targeted anticancer drug delivery.

GFP1, a sulfated polysaccharide extracted from Grateloupia filicina, exhibits remarkable immunomodulatory activity. To reduce the side effects of 5-fluorouracil (5-FU), GFP1 was employed as a macromolecular carrier to synthesize of GFP1-C-5-FU by reacting with carboxymethyl-5-fluorouracil (C-5-FU). Subsequently, this new compound was reacted with folic acid (FA) through an ester bond, forming novel conjugates named GFP1-C-5-FU-FA. Nuclear magnetic resonance analysis confirmed the formation of GFP1-C-5-FU-FA. In vitro drug release studies revealed that the cumulative release rate of C-5-FU reached 46.9 % in phosphate buffer (pH 7.4) after 96 h, a rate significantly higher than that of the control groups, indicating the controlled drug release behavior of GFP1-C-5-FU-FA. Additionally, in vitro anticancer assays demonstrated the potent anticancer activity of GFP1-C-5-FU-FA conjugates, as evidenced by the reduced viability of HeLa and AGS cancer cells, along with increased levels of apoptosis and cellular uptake. Western blot analysis indicated that the GFP1-C-5-FU-FA conjugate effectively enhanced phosphorylation in cancer cells through the NF-kB and MAPK pathways, thereby promoting apoptosis. These findings highlight the potential of folate-targeted conjugates in efficiently treating HeLa and AGS cancer cells in vitro and lay a robust theoretical groundwork for future in vivo anti-cancer research involving these cells.