Current findings support the potential use of bioactive peptides in enhancing zinc absorption in humans.

More than two billion people around the world are affected by zinc deficiency, mainly due to the inadequate intake and absorption of zinc. Based on recent research findings, the bioactive peptides could potentially be used to combat zinc deficiency particularly due to their Zinc chelating ability. The main aim of this review was to present current findings, supporting the potential use of bioactive peptides based on their ability to enhance zinc absorption. In-vivo, in-vitro, and ex-vivo studies have demonstrated that zinc chelating peptides can enhance the retention, transportation, and absorption of zinc. Comparative studies on zinc bioavailability from protein hydrolysates and zinc salts have demonstrated that the protein hydrolysates-zinc complexes are more bioavailable than the zinc salts. Data from the structure-function relationship of zinc chelating peptides suggest that the zinc chelating capacities of peptides increase in the following order; the position of zinc chelator > zinc chelator strength > abundance of zinc chelators > net charge > molecular weight. In addition, the transport mechanism of peptide-zinc complex is hypothesized, and the potential use of bioactive peptides based on their safety and taste and limitations to their commercialization are also discussed.

Preparation, structural properties, and in vitro and in vivo activities of peptides against dipeptidyl peptidase IV (DPP-IV) and α-glucosidase: a general review.

Diabetes is one of the fastest-growing and most widespread diseases worldwide. Approximately 90% of diabetic patients have type 2 diabetes. In 2019, there were about 463 million diabetic patients worldwide. Inhibiting the dipeptidyl peptidase IV (DPP-IV) and α-glucosidase activity is an effective strategy for the treatment of type 2 diabetes. Currently, various anti-diabetic bioactive peptides have been isolated and identified. This review summarizes the preparation methods, structure-effect relationships, molecular binding sites, and effectiveness validation of DPP-IV and α-glucosidase inhibitory peptides in cellular and animal models. The analysis of peptides shows that the DPP-IV inhibitory peptides, containing 2-8 amino acids and having proline, leucine, and valine at their N-terminal and C-terminal, are the highly active peptides. The more active α-glucosidase inhibitory peptides contain 2-9 amino acids and have valine, isoleucine, and proline at the N-terminal and proline, alanine, and serine at the C-terminal.

Mathermycin, an anti-cancer molecule that targets cell surface phospholipids.

Mathermycin, a lantipeptide isolated from marine actinomycete Marinactinospora thermotolerans, is an antibiotic that has been shown to disrupt bacterial plasma membrane. We now provide evidences that mathermycin can also disrupt cancer, but not normal, cell plasma membranes through targeting phosphatidylethanolamine (PE), which is located only in the inner leaflet of the plasma membrane in normal cells but in both the inner and outer leaflets of the membrane in tumor cells. Our data shows that mathermycin inhibits the metabolic activity and induces mainly necrotic death of all cancer cell lines with EC50 between 4.2 and 16.9 µM, while normal cell lines have EC50 between 113 and 129 µM. The cytotoxicity of mathermycin could be inhibited by exogenous PE, but not phosphoserine and phosphocholine. The formation of mathermycin-PE complexes was confirmed by in silico analysis, HPLC and MS spectrometer. Furthermore, mathermycin exhibited similar cytotoxicity toward cancer and multidrug resistant cancer cells, which could be due to its ability to inhibit mitochondrial function, as shown by our data from the Seahorse™ metabolic analyzer. This study demonstrates that mathermycin is a potentially effective class of anti-tumor chemotherapeutics that do not easily develop resistance due to a mechanism of action targeting PE.

The Antidepressant-like Effect of Flavonoids from Trigonella Foenum-Graecum Seeds in Chronic Restraint Stress Mice via Modulation of Monoamine Regulatory Pathways.

Fenugreek (Trigonella Foenum-Graecum) seeds flavonoids (FSF) have diverse biological activities, while the antidepressant-like effect of FSF has been seldom explored. The aim of this study was to evaluate the antidepressant-like effect of FSF and to identify the potential molecular mechanisms. LC-MS/MS was used for the determination of FSF. Chronic restraint stress (CRS) was used to establish the animal model of depression. Observation of exploratory behavior in the forced swimming test (FST), tail suspension test (TST) and sucrose preference test (SPT) indicated the stress level. The serum corticosterone (CORT) level was measured. The monoamine neurotransmitters (5-HT, NE and DA) and their metabolites, as well as monoamine oxidase A (MAO-A) enzyme activity in the prefrontal cortex, hippocampus and striatum, were evaluated. The protein expression levels of KLF11, SIRT1, MAO-A were also determined by western blot analysis. The results showed that FSF treatment significantly reversed the CRS-induced behavioral abnormalities, including reduced sucrose preference and increased immobility time. FSF administration markedly restored CRS induced changes in concentrations of serum corticosterone, prefrontal cortex neurotransmitters (NE, 5-HT and DA), hippocampus neurotransmitters (NE, 5-HT and DA) and striatum neurotransmitters (NE). FSF treatment exhibited significant inhibition of MAO-A activity in the prefrontal cortex and hippocampus. FSF also significantly down-regulated the KLF11, SIRT1 and MAO-A protein expression levels in the prefrontal cortex and hippocampus. These findings indicate that FSF could exhibit an antidepressant-like effect by down-regulating the KLF11/SIRT1-MAO-A pathways, inhibiting MAO-A expression and activity, as well as up-regulating monoamine neurotransmitters levels.

Effects of Acanthopanax senticosus on Brain Injury Induced by Simulated Spatial Radiation in Mouse Model Based on Pharmacokinetics and Comparative Proteomics.

The active compounds in Acanthopanax senticosus (AS) have different pharmacokinetic characteristics in mouse models. Cmax and AUC of Acanthopanax senticosus polysaccharides (ASPS) were significantly reduced in radiation-injured mice, suggesting that the blood flow of mouse was blocked or slowed, due to the pathological state of ischemia and hypoxia, which are caused by radiation. In contrast, the ability of various metabolizing enzymes to inactivate, capacity of biofilm transport decrease, and lessening of renal blood flow accounts for radiation, resulting in the accumulation of syringin and eleutheroside E in the irradiated mouse. Therefore, there were higher pharmacokinetic parameters-AUC, MRT, and t1/2 of the two compounds in radiation-injured mouse, when compared with normal mouse. In order to investigate the intrinsic mechanism of AS on radiation injury, AS extract's protective effects on brain, the main part of mouse that suffered from radiation, were explored. The function of AS extract in repressing expression changes of radiation response proteins in prefrontal cortex (PFC) of mouse brain included tubulin protein family (α-, ß-tubulin subunits), dihydropyrimidinase-related protein 2 (CRMP2), γ-actin, 14-3-3 protein family (14-3-3ζ, ε), heat shock protein 90ß (HSP90ß), and enolase 2. The results demonstrated the AS extract had positive effects on nerve cells' structure, adhesion, locomotion, fission, and phagocytosis, through regulating various action pathways, such as Hippo, phagosome, PI3K/Akt (phosphatidylinositol 3 kinase/protein kinase B), Neurotrophin, Rap1 (Ras-related protein RAP-1A), gap junction glycolysis/gluconeogenesis, and HIF-1 (Hypoxia-inducible factor 1) signaling pathways to maintain normal mouse neurological activity. All of the results indicated that AS may be a promising alternative medicine for the treatment of radiation injury in mouse brain. It would be tested that whether the bioactive ingredients of AS could be effective through the blood-brain barrier in the future.

Protection of Spleen Tissue of γ-ray Irradiated Mice against Immunosuppressive and Oxidative Effects of Radiation by Adenosine 5'-Monophosphate.

The immune system is very sensitive to radiation. This study revealed that adenosine 5′-monophosphate (5′-AMP) increased the DNA contents of the spleen and the spleen index of irradiated mice. Moreover, the exogenous 5′-AMP could significantly repair the ultra-structure of the damaged spleen through transmission electron microscopy. When indicators of the mouse immune system were assessed, the flow cytometry and enzyme-linked immunosorbent assay (ELISA) revealed that the administration of exogenous 5′-AMP could reduce the apoptosis in the splenic cells. It could also regulate the transition of cells towards S phase, increase the proportion of CD4⁺ and CD8⁺ cellular subsets, and enhance the secretion of interleukin-2 (IL-2), IL-4, IL-10, and interferon-γ (IFN-γ). These effects were associated with a decrease in oxidative stress, as evidenced by changes in superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), catalase (CAT), reduced glutathione (GSH), and malondialdehyde (MDA) levels of spleen tissues. These results suggested that exogenous 5′-AMP could repair the damaged spleen, increase the spleen index, and regulate the cell cycles and apoptosis. There was an increase in the production of various cytokines and play a protective role on the immune system of irradiated mice by dynamically adjusting the REDOX balance.

Protective Effects on 60Co-γ Radiation Damage of Pine Cone Polyphenols from Pinus koraiensis-Loaded Chitosan Microspheres In Vivo.

A novel chitosan microsphere for encapsulating pine cone polyphenols (PP) from P. koraiensis was successfully prepared using an emulsion crosslinking technique. The characteristics of pine polyphenol-loaded microspheres (PPM) were determined using scanning electron microscopy (SEM) and a laser particle size detector. It was found that PPMs were spherical in shape with uniform particle size distribution patterns. The drug content and encapsulation rate of the microspheres were 7.47% and 73.6%, respectively, at a Ch/GA mass ratio of 0.7. The animal experiments showed that PPM had a stronger radiation protective effect than PP. PPM significantly increased the immune organ indices, the quantity of marrow DNA, the superoxide dismutase (SOD) activity, the splenocyte proliferation index, and the phagocytosis activity of monocytes. PPM also decreased the numbers of micronuclei in bone marrow cells and malondialdehyde (MDA) levels in plasma in mice exposed to 60Co γ-irradiation. In addition, gender differences in biological responses to exposure to radiation were observed.

Synergistic radiation protective effect of purified Auricularia auricular-judae polysaccharide (AAP IV) with grape seed procyanidins.

The aim of this study was to investigate the synergistic antioxidant potential and protective effect of grape seed procyanidins (GSP) in combination with Auricularia auricular-judae polysaccharides (AAP IV) on radiation injury in splenocytes. Rat splenocyte irradiation resulted in significantly higher apoptosis rate, malondialdehyde (MDA) (p < 0.005), reactive oxygen species (ROS) (p < 0.01); cell viability, total superoxide dismutase (T-SOD) (p < 0.01), catalase (CAT) (p < 0.01), glutathione peroxidase (GSH-PX) (p < 0.05), activity and glutathione (GSH) (p < 0.01) levels were significantly reduced, compared with the control group. "GSP + AAP IV" treatment of rat splenocytes at doses of "GSP (0.3 µg/mL) + AAP IV (50 µg/mL)" displayed higher radioprotective and antioxidative effects than the administration of either GSP or AAP IV, as evident by lower levels of MDA (p < 0.001) concentration, as well as higher cell viability and T-SOD (p < 0.05), CAT (p < 0.005), GSH-PX (p < 0.01) and GSH content compared to the radiation group. In addition, in vivo studies have shown that "GSP + AAP IV" significantly ameliorated the decrease of spleen index (p < 0.005) and spleen GSH (p < 0.005) levels and significantly inhibited the increase of MDA (p < 0.005) levels of spleen with radiation-induced damage, compared with the non-treated group. The in vivo and in vitro results suggested that GSP and AAP IV have a synergistic protective effect against radiation-induced injury by improving the antioxidant and immunomodulation activities.

Examining the wound healing potential of curcumin-infused electrospun nanofibers from polyglutamic acid and gum arabic.

Advancements in medicine have led to continuous enhancements and innovations in wound dressing materials, making them pivotal in medical care. We used natural biological macromolecules, γ-polyglutamic acid and gum arabic as primary raw materials to create nanofibers laden with curcumin by blending electrostatic spinning technology in the current investigation. These nanofibers were meticulously characterized using fluorescence microscopy, scanning electron microscopy, Fourier transform infrared (FTIR) spectroscopy, differential scanning calorimetry (DSC), and X-ray diffraction (XRD). Our comprehensive analyses confirmed the successful encapsulation of curcumin within the nanofiber carrier and it has uniform diameter, good water absorption and mechanical properties. Subsequently, we evaluated the antimicrobial effects of these curcumin-loaded nanofibers against Staphylococcus aureus through an oscillating flask method. We created a mouse model with acute full-thickness skin defects to further investigate the wound healing potential. We conducted various biochemical assays to elucidate the mechanism of action. The results revealed that curcumin nanofibers profoundly impacted wound healing. They bolstered the expression of TGF-ß1 and VEGF and reduced the expression of inflammatory factors, leading to an accelerated re-epithelialization process, enhanced wound contraction, and increased regeneration of new blood vessels and hair follicles. Furthermore, these nanofibers positively influenced the proportion of three different collagen types. This comprehensive study underscores the remarkable potential of curcumin-loaded nanofibers to facilitate wound healing and lays a robust experimental foundation for developing innovative, natural product-based wound dressings.

The effect of 5'-adenylic acid on hepatic proteome of mice radiated by 60Co γ-ray.

Understanding the protection mechanism of 5'-AMP requires comprehensive knowledge of the proteins expressed during the period that the body is exposed to irradiation. Proteomics provides the tools for such analyses. Here, the experimental ICR mice were divided into three groups (normal group, model group and 5'-AMP + irradiation group). After different treatment, the hepatic total protein of each animal in three groups was separated by two-dimensional gel electrophoresis (2-DE). 2-DE analysis revealed fifty-eight protein spots were differentially expressed in comparison to three groups. From 58 protein spots, we selected nine spots to identify by MALDI-TOF-MS and received credible results. They were determined to be type I arginase, annexin A5, regucalcin, catalase, Tpm3 protein, Pdia4 protein, 14-3-3 protein epsilon, NAD-Malate dehydrogenase and heat shock protein 90. Considering the characteristic of these proteins, we proposed a possible protection pathway.

Recent Advances in and Applications of Electrochemical Sensors Based on Covalent Organic Frameworks for Food Safety Analysis.

The international community has been paying close attention to the issue of food safety as a matter of public health. The presence of a wide range of contaminants in food poses a significant threat to human health, making it vital to develop detection methods for monitoring these chemical contaminants. Electrochemical sensors using emerging materials have been widely employed to detect food-derived contaminants. Covalent organic frameworks (COFs) have the potential for extensive applications due to their unique structure, high surface area, and tunable pore sizes. The review summarizes and explores recent advances in electrochemical sensors modified with COFs for detecting pesticides, antibiotics, heavy metal ions, and other food contaminants. Furthermore, future challenges and possible solutions will be discussed regarding food safety analysis using COFs.

Protective effect of anthocyanin from Lonicera Caerulea var. Edulis on radiation-induced damage in mice.

The radioprotective effect of anthocyanin extracted from Lonicera caerulea var. edulis (ALC), was studied in ICR mice. Different doses of ALC were intragastrically administered to mice once a day, prior to radiation. After two weeks, the mice received a one-time 5 Gy whole body (60)Coγ radiation. The spleen index, thymus index, activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px), malondialdehyde (MDA) content, and glutathione (GSH) content in liver tissue were measured. Compared with the radiation control group, the levels of MDA in all ALC treated groups decreased significantly (p < 0.05). Moreover, the GSH content, activities of SOD and GSH-Px in liver tissue were enhanced significantly (p < 0.05) in all ALC groups. These results demonstrate that ALC may be a potential radioprotector, and a further study of the molecular mechanism is needed for further application.

Chemical composition and antioxidant activities of three polysaccharide fractions from pine cones.

The traditional method of gas chromatography-mass spectrometry for monosaccharide component analysis with pretreatment of acetylation is described with slight modifications and verified in detail in this paper. It was then successfully applied to the quantitative analysis of component monosaccharides in polysaccharides extracted from the pine cones. The results demonstrated that the three pine cone polysaccharides all consisted of ribose, rhamnose, arabinose, xylose, mannose, glucose and galactose in different molar ratios. According to the recovery experiment, the described method was proved accurate and practical for the analysis of pine cone polysaccharides, meeting the need in the field of chemical analysis of Pinus plants. Furthermore; the chemical characteristics, such as neutral sugar, uronic acids, amino acids, molecular weights, and antioxidant activities of the polysaccharides were investigated by chemical and instrumental methods. The results showed that the chemical compositions of the polysaccharides differed from each other, especially in the content of neutral sugar and uronic acid. In the antioxidant assays, the polysaccharide fractions exhibited effective scavenging activities on ABTS radical and hydroxyl radical, with their antioxidant capabilities decreasing in the order of PKP > PAP > PSP. Therefore, although the polysaccharide fractions had little effect on superoxide radical scavenging, they still have potential to be developed as natural antioxidant agents in functional foods or medicine.

[Anti-tumor effect of phytic acid on human osteosarcoma U2OS cells in vitro].

OBJECTIVE: To explore the effect of phytic acid on the human osteosarcoma U20S cells and its mechanisms in vitro. METHODS: MTT assay was used to examine the cell proliferation. Scanning electron microscope (SEM) was used to observe the surface ultrastructure of U20S cells. Hoechst 33342 fluorescence staining assay were applied to study the pro-apoptosis effects. The immune histochemisty method was used to detect c-myc expression. RESULTS: Phytic acid could significantly inhibit the growth of U20S. The surface ultrastructure of U20S cells showed distinct morphological changes corresponding to a typical cellular surface morphology of apoptosis. From fluorescence staining, we observed chromoplasm pyknosis and apoptotic body. Expression of c-myc protein decreased. CONCLUSION: The anti-osteosarcoma mechanism of phytic acid may be related to its influence on inducing apoptosis and adjusting c-myc expression.

Portulaca oleracea L. Extract Alleviated Type 2 Diabetes Via Modulating the Gut Microbiota and Serum Branched-Chain Amino Acid Metabolism.

SCOPE: Portulaca oleracea L. extracts (PE) show hypoglycemic function, but the precise mechanism remains obscure. This study is designed to investigate the association of the antidiabetes effect of PE with the gut microbiota modulation and BCAAs metabolism. METHODS AND RESULTS: The Orbitrap LC-MS to Orbitrap Fusion Lumos Tribrid mass spectrometer is employed to analyze the major compounds in PE. The components of the intestinal microflora in diet-induced/STZ-treated diabetic mice are analyzed by high-throughput 16S rRNA genes sequencing. The results show that PE improves blood glucose and insulin level, increases anti-inflammatory cytokine level, lowers serum branched-chain amino acids (BCAAs), and increases serum glutamine level. PE also protects the mucosal epithelium of the colon and cecum from damage. On the impact of gut microbial composition, PE reduces the Firmicutes to Bacteroidetes ratio and the abundance of the Lachnospiraceae_NK4A136_group, Blautia, Ruminiclostridium_9, Dubosiella, and increases the abundance of the Bacteroides, Akkermansia, and Mucisprillum genera. Bacterial functionality prediction indicates PE potentially inhibits bacterial BCAAs biosynthesis, and promotes the tissue-specific expression of BCAAs catabolic enzyme for reducing BCAAs supplementation. CONCLUSION: These results reveal that PE improving T2D-related biochemical abnormalities is associated not only with gut microbiota modification but also with the tissue-specific expression of BCAAs catabolic enzyme.

Protective effects of polysaccharides from soybean meal against X-ray radiation induced damage in mouse spleen lymphocytes.

The aim of this study was to investigate radioprotective effect of the polysaccharides from soybean meal (SMP) against X-ray radiation-induced damage in mouse spleen lymphocytes. MTT and comet assay were performed to evaluate SMP's ability to prevent cell death and DNA damage induced by radiation. The results show that, X-ray radiation (30 KV, 10 mA, 8 min (4 Gy)) can significantly increase cell death and DNA fragmentation of mouse spleen lymphocytes. Pretreatment with SMP for 2 h before radiation could increase cell viability, moreover, the SMP can reduce X-ray radiation-induced DNA damage. The percentage of tail DNA and the tail moment of the SMP groups were significantly lower than those of the radiation alone group (p < 0.05). These results suggest SMP may be a good candidate as a radioprotective agent.

A novel melanin complex displayed the affinity to HepG2 cell membrane and nucleus.

Chitosan-melanin complex from Catharsius molossus L. has proven to possess superior pharmaceutical excipient performance and may be the new source of water-soluble protein-free natural melanin. Herein, it was enzymatically hydrolyzed into the chitooligosaccharide-melanin complex (CMC) whose main chemical units were composed of eumelanin and chitooligosaccharides and showed three-layer structures. Additionally, this biomacromolecule could self-assemble into 40 nm nanoparticles (CMC Nps) in a weakly acidic aqueous solution. Interestingly, CMC displayed strong affinity for cell membrane by binding the phosphatidylserine, glycoprotein, glycolipids and glycosaminoglycans accumulated on the surface of tumor cells, notably, CMC Nps could enter cells and mainly target the nucleus by interacting with DNA and/or RNA substrates located around the nucleus to disrupt the proliferation and apoptosis processes. The findings suggest CMC may be the novel material for subcellular organelle targeting of cancer cells.

[Growth inhibition and apoptosis-inducing effects of phytic acid in human gastric carcinoma cells].

OBJECTIVE: To explore the growth inhibition and apoptosis-inducing effects of phytic acid in human gastric cancer SGC-7901 cells. METHODS: The growth inhibition action of phytic acid on SGC-7901 cells was examined by MTT assay. AO/EB fluorescence staining and DNA ladder assay were applied to study the proapoptosis effects of phytic acid. The expression of apoptosis relative proteins, P53, were analyzed by using immune histochemisty method. RESULTS: Phytic acid treatment significantly inhibited the growth of human gastric cancer cell SGC-7901 and markedly caused their apoptosis following downregulation of P53 protein expression. CONCLUSION: The downregulation of apoptosis relative protein P53 expression was the possible mechanism of phytic acid induced growth inhibition and apoptosis in SGC-7901 cells.

Hepatoprotection of pine nut polysaccharide via NRF2/ARE/MKP1/JNK signaling pathways against carbon tetrachloride-induced liver injury in mice.

Previously, we obtained a purified polysaccharide (PNP40c-1) from Pinus koraiensis pine nut and reported its protective effect on carbon tetrachloride (CCl4)-induced liver injury in vitro. The object of this study is to investigate its hepatoprotective activity in vivo and elucidate the mechanism underlying the hepatoprotection. PNP40c-1 effectively prevented the accumulation of serum liver injury biomarkers including alanine aminotransferase, aspartate aminotransferase, alkaline phpsphatase and total bilirubin stimulated by CCl4. The pathological changes in PNP40c-1-treated mice livers were also markedly ameliorated. Results showed that PNP40c-1 suppressed the production of reactive oxygen species (ROS) and lipid peroxidation, upregulated Nrf2/ARE pathway and enhanced the antioxidant capacity of hepatocytes. Furthermore, the reaction between Nrf2 and ARE promoted the generation of Mkp1, which inhibited the activation of JNK induced by CCl4, and suppressed hepatocytes apoptosis by regulating the protein expression of Bax, cleaved-Caspase-3 and Bcl2, exerting hepatoprotective activity. Taken together, upregulation of Nrf2/ARE pathway and suppression of JNK activation via Nrf2/ARE/Mkp1/JNK signaling pathways are the main mechanisms underlying the hepatoprotective effect of PNP40c-1 against CCl4-induced mice liver injury. These results indicated that PNP40c-1 has potential to serve as a hepatoprotective agent against chemical induced hepatotoxicity.

Hepatoprotection mechanism against alcohol-induced liver injury in vivo and structural characterization of Pinus koraiensis pine nut polysaccharide.

Previous studies have shown that Pinus koraiensis pine nut polysaccharide PNP80b-2 exerts widely protective effects against liver injury induced by chemical pollutants, alcohol and drugs. By comparison, PNP80b-2 exhibits the strongest hepatoprotection against alcohol-induced liver injury (AILI). Thus, the purpose of this study is to investigate the hepatoprotection mechanisms of PNP80b-2 against AILI in vivo. The results indicated that PNP80b-2 alleviated oxidative stress induced by alcohol through enhancing antioxidant capacity of hepatocytes via NRF2/HO-1 pathway. PNP80b-2 also effectively suppressed the secretion of pro-inflammatory cytokines including TNF-α, IL-1ß and IL-6, exhibiting anti-inflammatory effects via NF-κB signaling pathway in AILI. In addition, PNP80b-2 protected mice from severe DNA damage induced by alcohol through regulating the expression of Hipk2, P53, Hp1γ and Wip1. Taken together all the results, PNP80b-2 exerts hepatoprotective activity against AILI in vivo through enhancing antioxidant capacity, suppressing inflammation response and promoting DNA damage repair in livers. Furthermore, the structural features of PNP80b-2 were also characterized. PNP80b-2, with molecular weight of 23.0 kDa, was found to be composed of 1,2-linked Galf, 1,2-linked Rhap, 1,4-linked Xylp, 1,6-linked Glcp, 1,4-linked GlcpA, 1,2,6-linked Galp, 1,4,6-linked Glcp, 1,2,3,4-linked Arap, 1-linked Galp and Leu- and Ile-linked O-glycopeptide bonds, based on the GC-MS and NMR results.