DEAE-cellulose Column Research Articles

Production and purification of protease produced by UV-mutated Pseudomonas aeruginosa

Background: The enzyme protease accounts for approximately two-thirds to one-third of all enzymes used in different industries. Protease is produced in large quantities as a crude enzyme for food industries. Objectives: The aim of this study was to investigate the effect of UV radiation on the protease produced by Pseudomonas aeruginosa. Methods: Of a total of 149 samples collected from various clinical cases (wounds, burns, otitis media, and urinary tract infections) in four hospitals in Baghdad for the isolation of P. aeruginosa, 90 isolates were identified as belonging to P. aeruginosa after isolation on a specific selective media, and the Vitek 2 system was later used for final identification. Thirty-six P. aeruginosa isolates were screened for protease production on a skim milk medium. Results: This study found that most of the P. aeruginosa isolates were protease producers and the isolate p3 was the best, which had a hydrolysis zone of up to 29 mm and their specific activity was 0.134 units (U)/mg protein in the culture filtrate. Therefore, this isolate was selected to study the effect of UV light on the production of the protease enzyme. The results showed that protease was produced at high levels from mutant p3 isolates after exposure to UV irradiation for different durations (15, 25, 35, 45, 55, 65, 75, 85, and 95 seconds). The specific activity of the enzyme in a crude filtrate was 0.65 U/mg protein compared with the wild type (0.134 U/mg protein). Protease produced from the mutated isolate p3 was later purified in three purification steps: first, its precipitate with 70% ammonium sulfate, second through a DEAE-cellulose column, the specific activity of which reached 7 U/mg protein, and in the final step through a Sephadex G-200 column. The purified enzyme protease had a specific activity that gradually increased to 10.5 U/mg with an 18.23-fold purification and 84.1% enzyme recovery. Conclusion: UV radiation was an efficient method to induce the production of protease enzymes.

Ultrasonic-assisted extraction, structural analysis and antioxidative mechanism of polysaccharide from sunflower disc

The crude sunflower disc polysaccharide (SDP) was obtained by using ultrasonic assist hot water extraction. Homogeneous SDP was separated by DEAE-cellulose column and Sephacryl S-400 HR column. Its physical properties and antioxidant mechanism was studied. Results showed that extraction yield of SDP was 15.3 %, total sugar was 91.3 %, weight molecular weight (Mw) was7.46 × 103 Da, number-average molecular weight (Mn) was 6.16 × 103 Da and polydispersion coefficient (Mw/Mn) was 1.21. SDP was composed of galactose (Gal), fucose (Fuc), xylose (Xyl), glucose (Glc), arabinose (Ara), rhamnose (Rha), glucuronic acid (Gle-UA) and galacturonic acid (Ga-UA), and the ratio of substance were 1.77∶2.00∶4.55∶8.44∶9.57∶14.26∶2.13∶57.28. Scanning electron microscopy (SEM) and X-ray diffraction (XRD) results showed that smooth surface and structure of SDP was relatively dense, and some crystallization existed, but the crystallization was weak. Two-dimensional structure of SDP showed that the main chain →4)-α-d-GalpA-(1→, →3)-α-l-Araf-(1→, →2)-α-l-Rhap-(1→, →2, 4)-α-l-Rhap-(1→, →2, 4)-α-d-GlcpA-(1→ and →3, 6)-β-d-Galp-(1→ interconnection form. Results of in vitro and in vivo of SDP and its derivatives showed that a good antioxidant activity. In vitro antioxidant activity, the scavenging rates of ABTS+, ·OH, DPPH and superoxide anions were 95.8 %, 93.9 %, 93.6 % and 92.1 %, respectively, when treated with 3.2 mg/mL of SDP solution. The preliminary antioxidative mechanism results showed that SDP had a regulatory effects on the down-regulation of Nrf2, HO-1 and NQO1 proteins in RAW264.7 cells. Preliminary pharmacokinetic analysis showed that the SDP could be absorbed orally and reached its peak concentration in the blood at about 2 h. The content of polysaccharide in blood reached the peak value at 30 min and reached the lowest value at 8 h after intravenous injection.

Antiviral activity of a polysaccharide from Sargassum fusiforme against respiratory syncytial virus

This experiment examined the antiviral activity of polysaccharides from Sargassum fusiforme against respiratory syncytial virus (RSV) in vitro, including their mechanism of action and preliminary structural analysis. Four polysaccharides (SFP1, SFP2, SFP3, and SFP4) were purified from Sargassum fusiforme using a DEAE-52 cellulose column and an NW Super 150 gel column. CCK-8 and western blot were utilized to study the antiviral activities and mechanisms of the polysaccharides. Preliminary structural analysis was conducted using HPLC and NMR techniques. The findings suggest that SFP4 (120 kD) is an acidic chemical compound composed of 88.8 % total sugars, 0.13 % proteins, 10.8 % glucuronidic acids, and 21.1 % sulfates. It contains at least ten monosaccharides, primarily mannuronic acid and fucose. Among the four polysaccharides, SFP4 had the highest anti-RSV activity, with a therapeutic index (TI) exceeding 139. SFP4 exhibited noteworthy antiviral efficacy in both upper and lower respiratory cells that were infected, especially when administered as a prophylactic treatment 2 h in advance. Furthermore, SFP4 showed a dose-dependent antiviral effect, with the highest therapeutic index (TI > 320) observed at a concentration of 7.81 μg·mL−1 during the prophylactic phase. It was speculated that SFP4's antiviral effect is due to its ability to inhibit the attachment of G-proteins to cells.

Purification and characterization of α-amylase from Acanthoscelides obtectus (Say) (Coleoptera: Chrysomelidae)

Acanthoscelides obtectus is one of the most notorious pests of stored kidney beans (Phaseolus vulgaris) worldwide. Kidney beans are an important source of food for these insects. α-Amylase is the main carbohydrate-digesting enzyme in animals including insects. In the current study, the biochemical analysis revealed higher α-amylase activity (U/ml) in 3rd and 4th larval instars but decreased gradually in subsequent developmental stages. However, the specific activity (U/mg) interestingly was highest in 1st instar and decreased in further developmental stages. During qualitative analysis of α-amylase using starch-agar and native PAGE, the maximum zone of starch lysis and a prominent band on the gel was observed in 3rd and 4th larval stages. The molecular mass of the native enzyme was also estimated and found to be 30.34 kDa. The crude α-amylase was further purified by ammonium sulfate precipitation, gel filtration on a Sephadex G-75, and ion exchange chromatography on the DEAE cellulose column. The purified amylase was found to be a monomer with a molecular mass of 15 kDa. The specific activity of the purified enzyme increased from 1.74 U/mg in the crude sample to 166.35 U/mg in the final purification step resulting in 95-fold purification with a yield of 11.14%. Further characterization of purified α-amylase revealed a pH optimum of 7.0 and a temperature optimum of 35 °C. Lineweaver-Burk plot analysis revealed Km and Vmax to be 0.09% and 3.3 U/mL, respectively. Oxalic acid, tannic acid, and HgCl2 significantly inhibited the enzyme, while the Na+, Ca++, and Mg++ ions acted as activators. In conclusion, the study revealed, the highest α-amylase activity in 3rd and 4th larval stages of Acanthoscelides obtectus followed by native and SDS PAGE resulting in molecular mass of 30.34 and 15 kDa respectively.

Structure identification and activity evaluation of polysaccharide from Codonopsis pilosula (C. pilosula nannf. Var. modesta (nannf.) L. T. Shen)

Codonopsis pilosula is a medicinal plant with properties related to food, and the antioxidant and hypoglycemic components of its natural polysaccharides, as well as their related mechanisms, have not been fully understood. This study examined the physicochemical properties and structure of C. pilosula polysaccharide (WCP) using an orthogonal extraction method involving hot water-alcohol precipitation, high-resolution spectroscopy, and microscopic imaging technology. The antioxidant and hypoglycemic activities of the polysaccharides were assessed. Six fractions (WCP1, WCP2, WCP3, WCP4, WCP5, and WCP6) were identified through purification using a DEAE-cellulose column and Sephadex G-100 gel column. Mannose, glucose, and arabinose were identified as the primary monosaccharide components of WCPs through Gas Chromatography (GC). The polysaccharide exhibited a crystal structure composed of irregularly entangled pyranose particles of WCPs as revealed by X-ray diffraction (XRD), scanning electron microscopy (SEM), and atomic force microscopy (AFM). The polysaccharide decomposed into smoother triple-helix fibrous flocs as indicated by Congo red analysis and circular dichroism analysis (CD). The antioxidant capacity of the polysaccharides against various free radicals was evaluated, with WCP4 demonstrating a significant reducing capacity. Moreover, WCP3 and WCP5 displayed notable inhibition rates of α-amylase and α-glucosidase, respectively. Molecular docking studies indicated the interaction of WCP5 with two enzymes, forming multiple hydrogen bonds through specific amino acids. In the gromacs simulation, both composite systems maintained a steady state in terms of root mean square deviation (RMSD), root mean square fluctuation (RMSF), and radius of gyration, providing strong support for their potential as a reference for the developing novel therapeutic approaches to diabetes.

Exploratory studies of the antidepressant effect of Cordyceps sinensis polysaccharide and its potential mechanism

Cordyceps sinensis, a traditionally prized medicinal fungus, contains polysaccharides as one of its main bioactive constituents, known for their significant immunomodulatory properties. In this study, we systematically investigated the composition and structure of Cordyceps sinensis polysaccharide, followed by an evaluation of its therapeutic effect on depression using a chronic restraint stress-induced depression model. The polysaccharide CSWP-2, extracted via hot water, precipitated with ethanol, and purified using DEAE-cellulose column chromatography from Cordyceps sinensis, is primarily composed of glucose, mannose, and galactose, with α-1,4-D-glucan as its major structural component. Behavioral tests, immunological profiling, metabolomics, and gut microbiota analyses indicated a notable ameliorative effect of CSWP-2 on depressive-like symptoms in mice. Furthermore, the action of CSWP-2 may be attributed to the modulation of the gut microbiome's abundance and its metabolic impacts, thereby transmitting signals to the host immune system and exerting immunomodulatory activity, ultimately contributing to its antidepressant effects.

Structural characterization and mast cell stabilizing activity of Red-edge tea polysaccharide

The potential anti-allergic properties of tea have been demonstrated in studies supporting theanine and catechin. However, research on tea polysaccharides' anti-allergic properties has been limited. In this study, we extracted red-edge tea crude polysaccharide (RETPS) and evaluated its anti-allergic activity using the mast cell, passive cutaneous anaphylaxis, and passive systemic anaphylaxis models. We purified RETPS using the DEAE-52 cellulose column, analyzed its composition and structural characteristics, and compared the anti-allergic properties of different polysaccharide fractions. The purified components RETPS-3 and RETPS-4 displayed higher galacturonic acid content and lower molecular weight (106.61 kDa and 53.95 kDa, respectively) compared to RETPS (310.54 kDa). In addition, RETPS-3 and RETPS-4 demonstrated superior anti-allergic activity than RETPS in mice's passive cutaneous and systemic allergic reactions. Our findings provide evidence of the anti-allergic potential of tea polysaccharides and offer a theoretical foundation for developing tea polysaccharides as a functional anti-allergic food product.

Ultrasound-assisted deep eutectic solvents extraction of polysaccharides from Loquat leaf: Process optimization and bioactivity study

Loquat leaves are the by-product of loquat fruit production. Polysaccharides are one of the main active ingredients in loquat leaves. In this study, polysaccharides were extracted from loquat leaves by ultrasonic-assisted deep eutectic solvents (DESs) extraction method. Further, the extracted crude loquat leaf polysaccharides (CLLP) were purified and separated via S-8 resin and DEAE-52 cellulose column chromatography, respectively. Additionally, the effects of polysaccharides on activity of sperm in boar semen preserved in medium at 17 °C, were evaluated preliminarily. DES, composed of choline chloride/ethylene glycol (1:6, molar ratio), was proved to be the suitable solvent for LLP extraction. The optimized extraction conditions were water content 44 %, liquid-solid ratio 1:29 (g/g), extraction temperature 61 °C and extraction time 98 min. Under these conditions, the LLP yield was 57.82 ± 1.50 mg/g. A homogeneous polysaccharide (LLP1-2, Mw: 2.17 × 104 Da) was isolated from CLLP. Its total sugar, uronic acid and protein contents were 76.31 ± 1.25 %, 14.19 ± 0.67 % and 3.28 ± 0.42 %, respectively. Further, 800 μg/mL LLP1-2 could effectively enhance the antioxidant activity of sperm. This study laid a foundation for DESs and column chromatography in the field of polysaccharide extraction and separation, proving that LLP can be used as a natural antioxidant for sperm preservation.

Effects of Separation and Purification Methods on Antioxidation, Hypoglycemic and DNA Protection Activity of Fenugreek Polysaccharide.

Six low molecular weight fenugreek polysaccharides (FP) were isolated and purified by ethanol stepwise precipitation (EFP-20, EFP-40, and EFP-60) and DEAE-52 cellulose column method (DFP-0, DFP-0.15, and DFP-0.3), respectively. The effects of different separation and purification techniques on the preliminary properties and biological activities of fenugreek polysaccharides were compared. The results showed that the DEAE-52 cellulose-eluted fractions had a higher total sugar content and displayed a looser structure. The molecular weights of all six fractions were in the range of 4-19 kDa, with significant changes in the ratio of galactose to mannose. All six fractions contained α-D-galactopyranose and β-D-mannopyranose structures. Activity tests showed that all six fractions possessed antioxidant, hypoglycemic and DNA-protective activities. Among them, the DFP-0 fraction showed the highest activity. Overall, different isolation and purification methods lead to changes in the properties and bioactivities of FP, which provides a theoretical basis for the development and application of FP in functional foods and drugs.

Structural characterization of Astragalus polysaccharide-D1 and its improvement of low-dose metformin effect by enriching Staphylococcus lentus

To explore the adjuvant therapy drugs of low-dose metformin, one homogeneous polysaccharide named APS-D1 was purified from Astragalus membranaceus by DEAE-52 cellulose and Sephadex G-100 column chromatography. Its chemical structure was characterized by molecular weight distribution, monosaccharide composition, infrared spectrum, methylation analysis, and NMR. The results revealed that APS-D1 (7.36 kDa) consisted of glucose, galactose, and arabinose (97.51 %:1.56 %:0.93 %). It consisted of →4)-α-D-Glcp-(1→ residue backbone with →3)-β-D-Galp-(1→ residue and terminal-α/β-D-Glcp-(1→ side chains. APS-D1 could significantly improve inflammation (TNF-α, LPS, and IL-10) in vivo. Moreover, APS-D1 improved the curative effect of low-dose metformin without adverse events. APS-D1 combined with low-dose metformin regulated several gut bacteria, in which APS-D1 enriched Staphylococcus lentus to produce l-carnitine (one of 136 metabolites of S. lentus). S. lentus and l-carnitine could improve diabetes, and reduction of S. lentusl-carnitine production impaired diabetes improvement. The combination, S. lentus, and l-carnitine could promote fatty acid oxidation (CPT1) and inhibit gluconeogenesis (PCK and G6Pase). The results indicated that APS-D1 enhanced the curative effect of low-dose metformin to improve diabetes by enriching S. lentus, in which the effect of S. lentus was mediated by l-carnitine. Collectively, these findings support that low-dose metformin supplemented with APS-D1 may be a favorable therapeutic strategy for type 2 diabetes.

Anti-Photodamage Effect of Agaricus blazei Murill Polysaccharide on UVB-Damaged HaCaT Cells.

UVB radiation is known to induce photodamage to the skin, disrupt the skin barrier, elicit cutaneous inflammation, and accelerate the aging process. Agaricus blazei Murill (ABM) is an edible medicinal and nutritional fungus. One of its constituents, Agaricus blazei Murill polysaccharide (ABP), has been reported to exhibit antioxidant, anti-inflammatory, anti-tumor, and immunomodulatory effects, which suggests potential effects that protect against photodamage. In this study, a UVB-induced photodamage HaCaT model was established to investigate the potential reparative effects of ABP and its two constituents (A1 and A2). Firstly, two purified polysaccharides, A1 and A2, were obtained by DEAE-52 cellulose column chromatography, and their physical properties and chemical structures were studied. A1 and A2 exhibited a network-like microstructure, with molecular weights of 1.5 × 104 Da and 6.5 × 104 Da, respectively. The effects of A1 and A2 on cell proliferation, the mitochondrial membrane potential, and inflammatory factors were also explored. The results show that A1 and A2 significantly promoted cell proliferation, enhanced the mitochondrial membrane potential, suppressed the expression of inflammatory factors interleukin-1β (IL-1β), interleukin-8 (IL-8), interleukin-6 (IL-6), and tumor necrosis factor α (TNF-α), and increased the relative content of filaggrin (FLG) and aquaporin-3 (AQP3). The down-regulated JAK-STAT signaling pathway was found to play a role in the response to photodamage. These findings underscore the potential of ABP to ameliorate UVB-induced skin damage.

Characterization and optimization of exopolysaccharide extracted from a newly isolated halotolerant cyanobacterium, Acaryochloris Al-Azhar MNE ON864448.1 with antiviral activity

Several antiviral agents lost their efficacy due to their severe side effects and virus mutations. This study aimed to identify and optimize the conditions for exopolysaccharide (EPS) production from a newly isolated cyanobacterium, Acaryochloris Al-Azhar MNE ON864448.1, besides exploring its antiviral activity. The cyanobacterial EPS was purified through DEAE-52 cellulose column with a final yield of 83.75%. Different analysis instruments were applied for EPS identification, including Fourier-transform infrared (FT-IR) spectroscopy, thermogravimetric analysis (TGA), and gas chromatographic-mass spectrometry (GC–MS). Plackett–Burman’s design demonstrated that working volume (X1), EDTA (X2), inoculum size (X3), CaCl2 (X4), and NaCl (X5) are the most important variables influencing EPS production. Central composite design (CCD) exhibited maximum EPS yield (9.27 mg/mL) at a working volume of 300 mL in a 1 L volumetric flask, EDTA 0.002 g/L, inoculum size 7%, CaCl2 0.046 g/L, and NaCl 20 g/L were applied. EPS showed potent antiviral activities at different stages of herpes simplex virus type-1 and 2 (HSV-1, HSV-2), adenovirus (ADV) and coxsackievirus (A16) infections. The highest half-maximal inhibitory concentration (IC50) (6.477 µg/mL) was recorded during HSV-1 internalization mechanism, while the lowest IC50 (0.005669 µg/mL) was recorded during coxsackievirus neutralization mechanism.

Purple sweet potato polysaccharide ameliorates concanavalin A-induced hepatic injury by inhibiting inflammation and oxidative stress

BackgroundAutoimmune hepatitis (AIH) is a prevalent liver disease that can potentially lead to hepatic fibrosis and cirrhosis. The prolonged administration of immunosuppressive medications carries significant risks for patients. Purple sweet potato polysaccharide (PSPP), a macromolecule stored in root tubers, exhibits anti-inflammatory, antioxidant, immune-enhancing, and intestinal flora-regulating properties. Nevertheless, investigation into the role and potential mechanisms of PSPP in AIH remains notably scarce. PurposeOur aim was to explore the possible protective impacts of PSPP against concanavalin A (Con A)-induced liver injury in mice. MethodsPolysaccharide was isolated from purple sweet potato tubers using water extraction and alcohol precipitation, followed by purification through DEAE-52 cellulose column chromatography and Sephadex G-100 column chromatography. A highly purified component was obtained, and its monosaccharide composition was characterized by high performance liquid chromatography (HPLC). Mouse and cellular models induced by Con A were set up to investigate the impacts of PSPP on hepatic histopathology, apoptosis, as well as inflammation- and oxidative stress-related proteins in response to PSPP treatment. ResultsThe administration of PSPP significantly reduced hepatic pathological damage, suppressed elevation of ALT and AST levels, and attenuated hepatic apoptosis in Con A-exposed mice. PSPP was found to mitigate Con A-induced inflammation by suppressing the TLR4-P2X7R/NLRP3 signaling pathway in mice. Furthermore, PSPP alleviated Con A-induced oxidative stress by activating the PI3K/AKT/mTOR signaling pathway in mice. Additionally, PSPP demonstrated the ability to reduce inflammation and oxidative stress in RAW264.7 cells induced by Con A in vitro. ConclusionPSPP has the potential to ameliorate hepatic inflammation via the TLR4-P2X7R/NLRP3 pathway and inhibit hepatic oxidative stress through the PI3K/AKT/mTOR pathway during the progression of Con A-induced hepatic injury. The results of this study have unveiled the potential hepatoprotective properties of purple sweet potato and its medicinal value for humans. Moreover, this study serves as a valuable reference, highlighting the potential of PSPP-1 as a drug candidate for the treatment of immune liver injury.

Inhibition of Proliferation and Induction of Apoptosis in Prostatic Carcinoma DU145 Cells by Polysaccharides from Yunnan Rosa roxburghii Tratt.

This study aimed to investigate methodologies for the extraction and purification of polysaccharides from Rosa roxburghii Tratt fruits and their impact on various cellular processes in prostate cancer DU145 cells, including survival rate, migration, invasion, cell cycle, and apoptosis. Compared to the control group, the polysaccharide exhibited a significant reduction in the viability, migration, and invasion rates of DU145 cells in a time- and dose-dependent manner within the polysaccharide-treated groups. Additionally, it effectively arrested the cell cycle of DU145 cells at the G0/G1 phase by downregulating the expressions of CDK-4, CDK-6, and Cyclin D1. Furthermore, it induced apoptosis by upregulating the expressions of Caspase 3, Caspase 8, Caspase 9, and BAX. Polysaccharides were extracted from Rosa roxburghii Tratt sourced from Yunnan, China. Extraction and decolorization methods were optimized using response surface methodology, based on a single-factor experiment. Polysaccharide purification was carried out using DEAE-52 cellulose and Sephadex G-100 column chromatography. The optimal dosage of R. roxburghii Tratt polysaccharide affecting DU145 cells was determined using the CCK-8 assay. Cell migration and invasion were assessed using transwell and scratch assays. Flow cytometry was employed to analyze the effects on the cell cycle and apoptosis. Western blotting and Quantitative real-time PCR were utilized to examine protein and mRNA expressions in DU145 cells, respectively. Rosa roxburghii Tratt polysaccharides, consisting of D-mannose, L-rhamnose, N-acetyl-D-glucosamine, D-galacturonic acid, D-glucose, D-galactcose, D-xylose, L-arabinose, and L-fucose, possess the ability to hinder DU145 cell proliferation, migration, and invasion while inducing apoptosis through the modulation of relevant protein and gene expressions.

Production of an Efficient Enzymatically Fab Fragment Antivenom against Cobra Snake (Naja naja oxiana) Venom.

Since around 100 years ago, the best treatment for millions of global snakebite victims has been polyclonal antivenoms. However, common antivenoms need continuous improvement to reduce rare, their side effects and get better performance. In the present study, Fab antivenom was produced through papain digestion of anti-cobra venom plasma, multi-step purification, and optimization, including ammonium sulfate precipitation and DEAE-cellulose column chromatography. Then, the existence of the corresponding Fab fragment antibody was seen and confirmed by SDS-PAGE method and double immunodifusion (Ouchterlony) test. In addition, the potency test in NIH laboratory mice revealed that each milliliter of the new Fab antivenom was able to neutralize 624 micrograms and (80LD50) of cobra venom, which is about 15% more efficient than the primary plasma of the same concentration, and 1.57 times more effective than the cobra antivenom found in commercial hexavalent antivenom of Razi Institute. According to the findings, it seems that this new Fab antivenom can be used as a new candidate for treatment of cobra snakebite victims.

Purification and Structural Characterization of Polysaccharides from Polygonum multiflorum Thunb. and Their Immunostimulatory Activity in RAW264.7 Cells.

Polygonum multiflorum Thunb. (PM) and derived products are broadly utilized in Chinese traditional medicine. According to our previous research, PM mostly contains polysaccharides, which display a wide range of biological activities. Two water-soluble polysaccharides (PMPs-1 and PMPs-2) were obtained from PM by DEAE-Cellulose and Sephadex G-100 column chromatography. Colorimetry, HPGPC-MALLS-RID, HPLC-PDA, methylation, FT-IR, NMR, and SEM were used to characterize these polysaccharides. PMPs-1 and PMPs-2 had average molecular weights of 255.5 and 55.7 kDa, respectively. PMPs-1 consisted of Man, Glc, Gal, and Ara at 0.9:78.6:1.0:1.6 and was a glucan with → 4)-Glcp-(1 → as a backbone. Meanwhile, PMPs-2, an acidic polysaccharide, comprised Rha, GalA, Glc, Gal, and Ara at 3.2:20.3:2.7:1.0:8.3. PMPs-1 and PMPs-2 significantly improved the proliferation of RAW 264.7 cells and induced NO, TNF-α, and IL-6 release. This study reveals that these two polysaccharides can be explored as novel immunomodulators and provide a basis for further development of PM in food and pharmaceutical industries.

Insight into antioxidant and anti-inflammatory effects of marine bacterial natural exopolysaccharide (EPSSM) using carrageenan-induced paw edema in rats

Inflammation is a part of the body’s intricate biological reaction to noxious stimuli and defensive reactions. So, the aim of this investigation was to study the anti-inflammatory activity of exopolysaccharide (EPSSM) using carrageenan-induced paw edema in rats. A halophilic bacterial strain was isolated from marine sediments in the Red Sea in Egypt. The isolate has been visually and physiologically recognized, as well as by analyzing its 16S rRNA gene, which confirms Kocuria sp. clone Asker4. This particular isolate can be referenced using the accession number OL798051.1. EPSSM was subjected to purification and fractionation by a DEAE-cellulose column. Preliminary chemical analysis of EPSSM indicated that the monosaccharides were fructose, glucuronic acid, and xylose, with 2.0, 0.5, and 1.0, respectively. The antioxidant potential of EPSSM was investigated, and it was discovered that the level of activity increased independently of the concentrations, reaching a maximum threshold of 94.13% at 100 µg/mL of EPSSM for 120 min. Also, EPSSM at 50 mg/kg orally produced a significant anti-inflammatory effect on the carrageenan model at 2, 3, and 4 intervals. The EPSSM intervention resulted in reductions in the levels of catalase and superoxide dismutase enzymes, as well as a decrease in glutathione. Furthermore, the levels of nitric oxide, lipid peroxidation, and reactive oxygen species resulting from carrageenan-induced edema showed a significant reduction subsequent to the administration of EPSSM. Moreover, the findings indicated that the protein expression levels of cyclooxygenase-2 and interleukin-6 were reduced following treatment with EPSSM, resulting in a reduction of paw edema.

Isolation, Purification, Fractionation, and Hepatoprotective Activity of Polygonatum Polysaccharides.

In this study, three homogeneous fractions, PSP-N-b-1, PSP-N-b-2, and PSP-N-c-1, were obtained from an aqueous extract of Polygonatum using DEAE cellulose column chromatography, CL-6B agarose gel chromatography, and Sephadex G100 chromatography. Their monosaccharide compositions and molecular weights were analyzed. The results revealed that PSP-N-b-1, PSP-N-b-2, and PSP-N-c-1 are primarily composed of six monosaccharides: Man (mannose), GlcA (glucuronic acid), Rha (rhamnose), GalA (galacturonic acid), Glc (glucose), and Ara (arabinose), with molecular weights of 6.3 KDa, 5.78 KDa, and 3.45 KDa, respectively. Furthermore, we observed that Polygonatum polysaccharides exhibited protective effects against CCL4-induced liver damage in HepG2 cells in vitro, operating through both anti-oxidant and anti-inflammatory mechanisms. Our research findings suggest that Polygonatum polysaccharides may emerge as a promising option in the development of hepatoprotective drugs or functional foods with anti-inflammatory and antioxidant properties.

Synovial Fluid Proteomics and Serum Metabolomics Reveal Molecular and Metabolic Changes in Osteoarthritis

Background: Osteoarthritis (OA) is a common joint disorder with a complex and multifactorial pathogenesis. Proteomics analysis using two-dimensional gel electrophoresis (2DE) and mass spectrometry (MS) enables high-throughput identification of differentially expressed proteins related to OA. However, the etiology, pathophysiology, and early diagnostic markers of OA are still poorly understood. Methods: Synovial fluid protein biomarkers were compared between OA patients and healthy controls. It was fractionated using DEAE cellulose and Sephadex G-200 columns, followed by SDS‒PAGE and 2D-PAGE for visualization and identification. Mass spectrometry and Mascot were used for protein analysis, and serum metabolite profiles were also investigated using 1D 1H CPMG NMR spectra. Multivariate data analysis, including PCA and PLS-DA, was performed to detect metabolic differences between groups. Results: Proteomics analysis revealed differential expression of synovial fluid proteins, such as serine protease inhibitors, complement components, and apolipoproteins, which may be involved in inflammation and cartilage breakdown. Additionally, serum metabolite profiles differed significantly between OA patients and controls, involving amino acid, lipid, glucose, and energy metabolism. The pathway analysis indicated disruption of the metabolic pathways associated with these metabolites. Conclusions: This study provides insights into the molecular and metabolic changes in OA. Protein biomarkers and serum metabolite alterations enhance the understanding of OA pathogenesis and offer potential opportunities for early diagnosis and disease management. Further validation and translation of these findings into clinical applications are needed for improved OA detection and intervention strategies.

Potential osteoporosis-blocker Sparassis crispa polysaccharide: Isolation, purification and structure elucidation

This study aimed to investigate the structural characterization of water-soluble polysaccharides from Sparassis crispa and their effects on the proliferation and differentiation of mouse osteoblasts. Three fractions (F-1, F-2, and F-3) were obtained from crude polysaccharides by a DEAE-52 cellulose column. The main fraction (F-1) was further purified by polysaccharide gel purification systems to obtain purified water-soluble Sparassis crispa polysaccharide (SCPS). The chemical structure of SCPS was analyzed by gas chromatography, Fourier transform infrared spectroscopy, methylation analysis, and nuclear magnetic resonance spectroscopy. The monosaccharide compositional analysis revealed that SCPS consisted of fucose, arabinose, galactose, glucose, xylose, mannose, ribose, galacturonic acid, glucuronic acid, and mannuronic acid in a molar ratio of 17.37:1.94:25.52:30.83:1.14:0.30:4.98:2.87:2.65. Moreover, the backbone of SCPS was composed of →3)-β-d-Glcp-(1→4)-β-d-Glcp-(1→, with side chains attached to the backbone at the O-6 positions through the →3,6)-β-d-Glcp-(1→ linkage. The in vitro experiments were conducted to investigate the effects of SCPS on the proliferation and differentiation of mouse osteoblasts. The results showed that SCPS significantly enhanced the proliferation and differentiation of mouse osteoblasts, indicating their potential as a pharmaceutical agent for promoting osteoblast proliferation and differentiation.