Real-Time Monitoring of Translocation, Dissipation, and Cumulative Risk of Maleic Hydrazide in Potato Plants and Tubers by Ion Exclusion Chromatography.

In this paper, a high-performance ion exclusion chromatographic (ICE) method was developed and applied for monitoring maleic hydrazide (MH) translocation in complex potato plant tissue and tuber matrices. After middle leaf uptake, most MH was trapped and dissipated in the middle leaf, and the rest was transported to other parts mainly through the phloem. Soil absorption significantly reduced the uptake efficiency of the root system, in which MH was partitioned to dissipate in root protoplasts or transfer through the xylem and persisted in the plant. Tuber uptake enabled MH to remain in the flesh and maintain stable levels under storage conditions, but during germination, MH was translocated from the flesh to the growing buds, where it dissipated through the short-day photoperiodic regime. The results demonstrated successful application of the ICE method and provided necessary insights for real-time monitoring of MH translocation behavior to effectively improve potato edible safety.

Simultaneous Determination of Vitamin E and γ-Oryzanol in Rice Bran Oil via HPSEC-PDA without Sample Pretreatment.

Vitamin E (tocopherols and tocotrienols) and γ-oryzanol are two minor constituents of rice bran oil (RBO) and are known to be potential bioactive compounds. The content of γ-oryzanol, a unique antioxidant found only in RBO, is a key factor in determining the retail price of the oil. Limitations of conventional HPLC columns for vitamin E and γ-oryzanol analysis are the alteration of these components and the time-consuming need for pretreatment of samples by saponification. High-performance size exclusion chromatography (HPSEC) equipped with a universal evaporative light scattering detector (ELSD) is a versatile tool for screening optimum mobile phase conditions because components of the sample can be separated and detected in the same run. In this work, the RBO components (triacylglycerol, tocopherols, tocotrienols, and γ-oryzanol) assessed on a single 100-Å Phenogel column using ethyl acetate/isooctane/acetic acid (30:70:0.1, v/v/v) as the mobile phase provided baseline separations (R s >1.5) with a total run time of 20 min. The HPSEC condition was then transferred to determine the content of tocopherols, tocotrienols, and γ-oryzanol in RBO products using a selective PDA detector. The limit of detection (LOD) and limit of quantification (LOQ) of α-tocopherol, α-tocotrienol, and γ-oryzanol were 0.34 and 1.03 µg/mL, 0.26 and 0.79 µg/mL and 2.04 and 6.17 µg/mL, respectively. This method was precise and accurate, with a percentage of relative standard deviation (%RSD) of the retention time of less than 0.21%. The intra-day and inter-day variations were 0.15-5.05% and 0.98-4.29% for vitamin E and γ-oryzanol, respectively. The recoveries of tocopherols, tocotrienols, and γ-oryzanol ranged between 90.75% and 107.98%. Thus, the developed HPSEC-ELSD-PDA method is a powerful analytical tool for determining the vitamin E and γ-oryzanol present in oil samples without requiring any sample pretreatment.

Investigation of Best Practices for Venom Toxin Purification in Jellyfish towards Functional Characterisation.

The relative lack of marine venom pharmaceuticals can be anecdotally attributed to difficulties in working with venomous marine animals, including how to maintain venom bioactivity during extraction and purification. The primary aim of this systematic literature review was to examine the key factors for consideration when extracting and purifying jellyfish venom toxins to maximise their effectiveness in bioassays towards the characterisation of a single toxin.An up-to-date database of 119 peer-reviewed research articles was established for all purified and semi-purified venoms across all jellyfish, including their level of purification, LD50, and the types of experimental toxicity bioassay used (e.g., whole animal and cell lines). We report that, of the toxins successfully purified across all jellyfish, the class Cubozoa (i.e., Chironex fleckeri and Carybdea rastoni) was most highly represented, followed by Scyphozoa and Hydrozoa. We outline the best practices for maintaining jellyfish venom bioactivity, including strict thermal management, using the "autolysis" extraction method and two-step liquid chromatography purification involving size exclusion chromatography. To date, the box jellyfish C. fleckeri has been the most effective jellyfish venom model with the most referenced extraction methods and the most isolated toxins, including CfTX-A/B. In summary, this review can be used as a resource for the efficient extraction, purification, and identification of jellyfish venom toxins.

[Determination of 101 pesticide residues in Platycodonis radix and extracts of Angelica sinensis by gel permeation chromatography-gas chromatography-ion trap mass spectrometry].

With the growing global popularity of traditional medicine and natural drugs， especially in Southeast Asia， the quality of traditional Chinese medicines （TCMs） has attracted the attention of regulators. China's major TCM export destinations， such as South Korea， Japan， and Europe， have formulated strict maximum residue limits （MRLs） of pesticides in TCMs. Therefore， a sensitive and high-throughput method for the simultaneous determination of 101 pesticide residues in Platycodonis radix and extracts of Angelica sinensis was established， involving gel permeation chromatography （GPC） coupled with gas chromatography-ion trap mass spectrometry （GC-ITMS）. In this method， the samples were first ground into fine powder and extracted twice with 20 mL acetonitrile in an ultrasonic cleaner for 30 min. After centrifugation for 10 min at 6000 r/min， the supernatants were combined and dried at 40 ℃ using a rotary vacuum evaporator. The residue was re-dissolved in 2 mL ethyl acetate-cyclohexane （1∶1， v/v） and purified by gel permeation chromatography using a 40 cm×20 mm column. The eluent collecting time was optimized as 17-30 min to ensure both the recovery of target compounds and the removal of interferences such as pigments and lipids from the target compounds. The eluent was then dried and re-dissolved with 1 mL toluene for analysis. The 101 pesticide residues were separated using the DB-5MS capillary column and analyzed by ion trap mass spectrometry. The pretreatment conditions and ion trap mass spectrometry parameters were optimized to effectively reduce the interference of complex TCM matrices and greatly improve the quantitative accuracy of the analysis and recovery of the target pesticides. Three spiked levels of 101 pesticides were tested. The average recovery range was 58.3%-108.9% and the relative standard deviations （RSDs） at the three spiked levels （n=10） ranged from 0.4% to 16.5%. The limits of detection （LODs， S/N=3） of the 101 pesticide compounds ranged from 0.2 to 40.0 µg/kg， while the limits of quantification （LOQs， S/N=10） ranged from 0.6 to 120.0 µg/kg， which met the maximum residue limits of China's main TCM export countries and organizations. This rapid analysis method was easy to operate and high throughput， with strong sensitivity and good repeatability. The employment of gel permeation chromatography overcame the drawback of inadequate cleanup of the solid phase extraction column during TCM analysis. The application of ion trap technology further eliminated the interference of matrix impurities and increased the accuracy of the quantitative and qualitative analyses. This method fills the knowledge gap in multiple pesticide residue determination in TCMs using gas chromatography-ion trap mass spectrometry and is a useful and beneficial alternative to current analytical methods of TCMs.

Fractional extraction and structural characterization of glycogen particles from the whole cultivated caterpillar fungus Ophiocordyceps sinensis.

Ophiocordyceps sinensis (syn. Cordyceps sinensis) is a valuable medicinal fungus in traditional Chinese medicine, and one or more polysaccharides are the key constituents with important medical effects. Glycogen as a functional polysaccharide is widely identified in eukaryotes including fungi. However, there is no definitive report of glycogen presence in O. sinensis. In this study, we carefully fractionated polysaccharides from cultivated caterpillar fungus O. sinensis, which were then characterized via methods for glycogen analysis. According to the results, 1.03 ± 0.43 % of polysaccharides were quantified via amyloglucosidase digestion in the whole cultivated caterpillar fungus, which had a typical spherical shape under transmission electron microscope with an average peak radius of 37.63 ± 0.57 nm via size exclusion chromatography and an average chain length of 12.47 ± 0.94 degree of polymerization via fluorophore-assisted capillary electrophoresis. Taken together, this study confirmed that the polysaccharides extracted form O. sinensis were mostly glycogen.

Isolation, Characterization of Bamboo Leaf Flavonoids by Size Exclusion Chromatography and Their Antioxidant Properties.

The utilization of bamboo industry exhibits varied but still needs to be improved. Bamboo leaf flavonoid (BLF) is an important resource of bamboo which has become a research focus. However, the isolation and purification techniques of four flavonoid carbon glycosides (orientin, isoorientin, vitexin, and isovitexin) from BLF were still confronted with difficulties due to their complex and similar structures, which obstructed the development of bamboo utilization. In this article, a purification technology of four flavonoid carbon glycosides from BLF by Sephadex LH-20 was improved. The results were evaluated by HPLC and pharmacological activity. Specifically, the eluent, flow rate, and loading amount were investigated, respectively. According to the results, the eluent would dominate the isolation effect among three factors. High concentration of isoorientin and four flavonoid carbon glycosides would be obtained under the optimized condition (The eluent was 70 % methanol, the loading amount was 1.5 g, and the flow rate was 0.5 mL/min). Meanwhile, the link between flavonoid carbon glycosides content and their antioxidant activity in vitro was also revealed. Overall, the results suggested that BLF may serve as potential functional food additives and medicine.

Understanding the structure and composition of recalcitrant oligosaccharides in hydrolysate using high-throughput biotin-based glycome profiling and mass spectrometry.

Novel Immunological and Mass Spectrometry Methods for Comprehensive Analysis of Recalcitrant Oligosaccharides in AFEX Pretreated Corn Stover. Lignocellulosic biomass is a sustainable alternative to fossil fuel and is extensively used for developing bio-based technologies to produce products such as food, feed, fuel, and chemicals. The key to these technologies is to develop cost competitive processes to convert complex carbohydrates present in plant cell wall to simple sugars such as glucose, xylose, and arabinose. Since lignocellulosic biomass is highly recalcitrant, it must undergo a combination of thermochemical treatment such as Ammonia Fiber Expansion (AFEX), dilute acid (DA), Ionic Liquid (IL) and biological treatment such as enzyme hydrolysis and microbial fermentation to produce desired products. However, when using commercial fungal enzymes during hydrolysis, only 75-85% of the soluble sugars generated are monomeric sugars, while the remaining 15-25% are soluble recalcitrant oligosaccharides that cannot be easily utilized by microorganisms. Previously, we successfully separated and purified the soluble recalcitrant oligosaccharides using a combination of charcoal and celite-based separation followed by size exclusion chromatography and studies their inhibitory properties on enzymes. We discovered that the oligosaccharides with higher degree of polymerization (DP) containing methylated uronic acid substitutions were more recalcitrant towards commercial enzyme mixtures than lower DP and neutral oligosaccharides. Here, we report the use of several complementary techniques that include glycome profiling using plant biomass glycan specific monoclonal antibodies (mAbs) to characterize sugar linkages in plant cell walls and enzymatic hydrolysate, matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS) using structurally-informative diagnostic peaks offered by negative ion post-secondary decay spectra, gas chromatography followed by mass spectrometry (GC-MS) to characterize oligosaccharide sugar linkages with and without derivatization. Since oligosaccharides (DP 4-20) are small, it is challenging to mobilize these molecules for mAbs binding and characterization. To overcome this problem, we have applied a new biotin-coupling based oligosaccharide immobilization method that successfully tagged most of the low DP soluble oligosaccharides on to a micro-plate surface followed by specific linkage analysis using mAbs in a high-throughput system. This new approach will help develop more advanced versions of future high throughput glycome profiling methods that can be used to separate and characterize oligosaccharides present in biomarkers for diagnostic applications.

Structural characterisation and bioactivity of polysaccharides isolated from fermented Dendrobium officinale.

BACKGROUND: A polysaccharide was purified in this study, which was acquired from the fermentation broth of Dendrobium officinale Kimura et Migo. We aimed to investigate the structural features and bioactivity of this polysaccharide. RESULTS: The polysaccharide was purified and the main polysaccharide fraction (i.e., DOP-1) was obtained. High-performance gel permeation chromatography (HPGPC) revealed that the molecular weight of DOP-1 was 447.48 kDa. Galactose, glucose and mannose were found to be present in DOP-1 via monosaccharide composition analysis, at a ratio of 1:1.79:6.71. Methylation and nuclear magnetic resonance spectroscopic analysis indicated that the backbone of DOP-1 was →4)-α-d-Glcp-(1 → 4)-α-d-Manp-(1 → 4)-α-d-Manp-(1 → 4,6)-α-d-Manp-(1→, and its repeating units were also preliminarily established. In vitro tests proved that DOP-1 not only protects RAW264.7 macrophages from the cytotoxic effect induced by lipopolysaccharide (LPS), but also inhibits cytokines (i.e., interleukin-6 and tumour necrosis factor-α) induced by LPS. DOP-1 demonstrated good scavenging activity in vitro toward 1,1-diphenyl-2-picrylhydrazyl and hydroxyl radicals, as well as good metal chelating activity. Therefore, DOP-1 has potential antioxidant applications. CONCLUSION: The structural characteristics of DOP-1 support its favourable biological activities and lay a strong foundation for further exploration of its structure-activity relationships and activity development, providing experimental data for the development and utilisation of fermentation broth of D. officinale. © 2021 Society of Chemical Industry.

New diketopiperazine alkaloid and polyketides from marine-derived fungus Penicillium sp. TW58-16 with antibacterial activity against Helicobacter pylori.

Marine-derived fungi can usually produce structurally novel and biologically potent metabolites. In this study, a new diketopiperazine alkaloid (1) and two new polyketides (10 and 11), along with 8 known diketopiperazine alkaloids (2-9) were isolated from marine-derived fungus Penicillium sp. TW58-16. Their structures were fully elucidated by analyzing UV, IR, HR-ESI-MS, 1D, and 2D NMR spectroscopic data. The absolute configurations of the new compounds 1, 10 and 11 were ascertained by X-ray diffraction (Cu Kα radiation) and comparing their CD data with those reported. In addition, the antibacterial activities of these compounds against Helicobacter pylori in vitro were assessed. Results showed that compounds 3, 6, 8 and 9 displayed moderate antibacterial activity against standard strains and drug-resistant clinical isolates of H. pylori in vitro. This result demonstrates that diketopiperazine alkaloids could be lead compounds to be explored for the treatment of H. pylori infection.

Characterizing the Diffusion and Rheological Properties of Aged Asphalt Binder Rejuvenated with Bio-Oil Based on Molecular Dynamic Simulations and Laboratory Experimentations.

Soybean-derived bio-oil is one of the vegetable-based oils that is gaining the most interest for potential use in the rejuvenation of aged asphalt binders. This laboratory study was conducted to characterize and quantify the diffusion and rheological properties of bio-oil-rejuvenated aged asphalt binder (BRAA) using soybean oil. In the study, the chemical structure of the soybean oil was comparatively characterized using an element analyzer (EA), gel permeation chromatography (GPC), and a Fourier infrared (FTIR) spectrometer, respectively. Based on the chemical structure of the bio-oil, BRAA molecular models were built for computing the diffusion parameters using molecular dynamic simulations. Likewise, a dynamic shear rheometer (DSR) test device was used for measuring and quantifying the rheological properties of the aged asphalt binder rejuvenated with 0%, 1%, 2%, 3%, 4%, and 5% soybean oil, respectively. The laboratory test results indicate that bio-oil could potentially improve the diffusion coefficients and phase angle of the aged asphalt binder. Similarly, the corresponding decrease in the complex shear modulus has a positive effect on the low-temperature properties of BRAA. For a bio-oil dosage 4.0%, the diffusion coefficients of the BRAA components are 1.52 × 10-8, 1.33 × 10-8, 3.47 × 10-8, 4.82 × 10-8 and 3.92 × 10-8, respectively. Similarly, the corresponding reduction in the complex shear modulus from 1.27 × 107 Pa to 4.0 × 105 Pa suggests an improvement in the low-temperature properties of BRAA. Overall, the study contributes to the literature on the potential use of soybean-derived bio-oil as a rejuvenator of aged asphalt binders.

Size exclusion chromatography of aromatic compounds in high-boiling petroleum samples.

Knowledge of aromatic compounds in petroleum samples is of paramount interest from processing and environmental viewpoints. Considering the complexity of such samples, a selective analytical strategy viz. aromatic specific size exclusion chromatographic methodology using amino-bonded silica phase, was adopted to obtain the average molecular weights of distilled cuts, residue and the corresponding crude oil. The method was found to be suitable from middle distillates up to the residue. It was found that the increase in boiling temperature of distilled cuts was reflected in an increase in high molecular weight aromatic species. The increase in the molecular weight of aromatic species is linked to the increase in alkylation as verified by studying the elution pattern of alkylated aromatic compounds. The accuracy of the molecular weights obtained from the size exclusion chromatographic method depends on the elution profile of aromatic species in petroleum samples. It was found that the size and shape of the parent aromatic compounds negatively influence the size exclusion elution profile. However, such influence can be assumed to be negligible as all the aromatic species of each distilled cut, residue, and crude oil eluted in the permeation zone of the chromatogram. This method has the potential to be used in the second dimension of a 2D-LC method where the aromatic compounds are separated based on the size (or alkylation). The first dimension will separate compounds based on the number of aromatic rings, thereby eliminating the interference of parent aromatic compounds of different sizes in the total elution profile.

Study of Anti-Inflammatory and Analgesic Activity of Scorpion Toxins DKK-SP1/2 from Scorpion Buthus martensii Karsch (BmK).

Buthus martensii Karsch (BmK), is a kind of traditional Chinese medicine, which has been used for a long history for the treatment of many diseases, such as inflammation, pain and cancer. In this study, DKK-SP1/2/3 genes were screened and extracted from the cDNA library of BmK. The DKK-SP1/2/3 were expressed by using plasmid pSYPU-1b in E. coli BL21, and recombinant proteins were obtained by column chromatography. In the xylene-induced mouse ear swelling and carrageenan-induced rat paw swelling model, DKK-SP1 exerted a significant anti-inflammatory effect by inhibiting the expression of Nav1.8 channel. Meanwhile, the release of pro-inflammatory cytokines (COX-2, IL-6) was decreased significantly and the release of anti-inflammatory cytokines (IL-10) were elevated significantly. Moreover, DKK-SP1 could significantly decrease the Nav1.8 current in acutely isolated rat DRG neurons. In the acetic acid-writhing and ION-CCI model, DKK-SP2 displayed significant analgesic activity by inhibiting the expression of the Nav1.7 channel. Moreover, DKK-SP2 could significantly inhibit the Nav1.7 current in the hNav1.7-CHO cells.

Structural characterization and osteogenic activity in vitro of novel polysaccharides from the rhizome of Polygonatum sibiricum.

Polygonatum sibiricum Red., a kind of edible and medicinal plant, has a long history of use in traditional Chinese medicine and as a nutritional food additive. Water-soluble crude polysaccharide (PS50) was obtained from P. sibiricum rhizome. After deproteinization and dialysis, PS50 was isolated and purified via DEAE-52 cellulose and Sephadex G-75 gel filtration chromatography to obtain two novel polysaccharides (PSP50-2-1 and PSP50-2-2). Chemical analysis and nuclear magnetic resonance (NMR) studies indicated that PSP50-2-1 was composed of ß-d-Glcp-(1→, →2)-ß-d-Galp-(1→, →2,6)-ß-d-Galp-(1→, ß-d-Fruf-(2→, and PSP50-2-2 was composed of ß-d-Glcp-(1→, ß-d-Galp-(1→, →2)-ß-d-Galp-(1→, →6)-ß-d-Galp-(1→, →2,6)-ß-d-Galp-(1→, ß-d-Fruf-(2→. The morphological analysis suggested that PSP50-2-1 and PSP50-2-2 both had a rough surface with cracks. Pharmacological studies showed that PSP50-2-2 at concentrations of 2.59 and 5.19 µM significantly promoted the differentiation and mineralization of MC3T3-E1 cells in vitro, which indicated that PSP50-2-2 had a function of osteogenic activity. Above all, this study provides evidence that PSP50-2-2 may be used as an anti-osteoporotic agent, with applications in health-care and pharmaceutical industries.

Isolation of sesquiterpenoids from Matricaria chamomilla by means of solvent assisted flavor evaporation and centrifugal partition chromatography.

The (semi)volatile fraction of Matricaria chamomilla L., an annual herbal plant from the family of Asteraceae, contains high quantities of sesquiterpenes and sesquiterpenoids. A method was developed to achieve isolation and separation of these compounds, using a combination of solvent assisted flavor evaporation (SAFE) and solid support-free liquid-liquid chromatography. The biphasic liquid solvent system n-heptane/ethyl acetate/methanol/water, 5/2/5/2 v/v/v/v (Arizona S) was elaborated as a suitable solvent system for the simultaneous separation of the target compounds. The lab-scale liquid-liquid chromatography separation performed in a countercurrent chromatography (CCC) column was successfully transferred to a semi-preparative centrifugal partition chromatography (CPC) column, which enabled the isolation of artemisia ketone, artemisia alcohol, α-bisabolone oxide A, and (E)-en-yn-dicycloether. α-Bisabolol oxide A and (Z)-en-yn-dicycloether co-eluted, but were successfully separated by subsequent size-exclusion chromatography (SEC). Similarly, spathulenol and α-bisabolol oxide B were obtained as a mixture, and were separated by means of column chromatography using silica gel as stationary phase. The isolated compounds were characterized by means of nuclear magnetic resonance spectroscopy (NMR) and gas chromatography-mass spectrometry (GC-MS).

Ultrasound treatment of crushed grapes: Effect on the must and red wine polysaccharide composition.

This paper studied the effect on the molecular weight and polysaccharide composition of musts and wines of the application of high-power ultrasound (US) at 20 and 28 kHz on crushed grapes. Two different pomace maceration times (short and mid) were tested for sonicated and control vinifications. A long pomace maceration time was also tested for non-treated wines. In must samples, US significantly increased the content of monosaccharides and polysaccharides rich in arabinose and galactose (PRAG), and the average molecular weight of smaller PRAG, mannoproteins (MP) or mannans. The 28 kHz had a major effect on most wine monosaccharides and grape polysaccharides. The wine obtained from sonicated grapes at 28 kHz and with mid maceration had higher rhamnogalacturonans type II and PRAG content than its control, and closer polysaccharide and monosaccharide content to long maceration control wines. No significant differences were obtained in the MP content between sonicated and control wines.

Biochemical characterization and enhanced production of endoxylanase from thermophilic mould Myceliophthora thermophila.

Endoxylanase production from M. thermophila BJTLRMDU3 using rice straw was enhanced to 2.53-fold after optimization in solid state fermentation (SSF). Endoxylanase was purified to homogeneity employing ammonium sulfate precipitation followed by gel filtration chromatography and had a molecular mass of ~ 25 kDa estimated by SDS-PAGE. Optimal endoxylanase activity was recorded at pH 5.0 and 60 °C. Purified enzyme showed complete tolerance to n-hexane, but activity was slightly inhibited by other organic solvents. Among surfactants, Tweens (20, 60, and 80) and Triton X 100 slightly enhanced the enzyme activity. The Vmax and Km values for purified endoxylanase were 6.29 µmol/min/mg protein and 5.4 mg/ml, respectively. Endoxylanase released 79.08 and 42.95% higher reducing sugars and soluble proteins, respectively, which control after 48 h at 60 °C from poultry feed. Synergistic effect of endoxylanase (100 U/g) and phytase (15 U/g) on poultry feed released higher amount of reducing sugars (58.58 mg/feed), soluble proteins (42.48 mg/g feed), and inorganic phosphate (28.34 mg/feed) in contrast to control having 23.55, 16.98, and 10.46 mg/feed of reducing sugars, soluble proteins, and inorganic phosphate, respectively, at 60 °C supplemented with endoxylanase only.

Integrated biological and chemical characterisation of a pair of leonardesque canal lock gates.

The Museo Nazionale della Scienza e della Tecnologia "Leonardo da Vinci" in Milan is exposing two pairs of canal lock gates, used to control the water flow in Milan canal system, whose design appears in the Leonardo's Codex Atlanticus. The wood present in the gates has been deeply characterised by mean of a multidisciplinary investigation involving i) DNA barcoding of wood fragments; ii) microbial community characterisation, and iii) chemical analyses. DNA barcoding revealed that two fragments of the gates belonged to wood species widely used in the middle age: Fagus sylvatica and Picea abies. The chemical characterisations were based on the use of ionic liquid as dissolving medium in order to analyse the entire cell wall material by means of Gel Permeation Chromatography (GPC) and 2D-NMR-HSQC techniques. This multidisciplinary analytical approach was able to highlight the complex nature of the degradation occurred during the gate operation (XVI-XVIII centuries): an intricate interplay between microbial populations (i.e. Shewanella), inorganic factors (i.e. iron from nails), physical factors and the lignocellulosic material.

Characterization of an antioxidant pectic polysaccharide from Platycodon grandiflorus.

Platycodonis Radix is widely used as homology of medicine and food in China; polysaccharides are thought to be one of its functional constituents. In this study, a pectic polysaccharide, PGP-I-I, was obtained from the root of the traditional medicine plant Platycodon grandiflorus through ion exchange chromatography and gel filtration. This was characterized being mainly composed of 1,5-α-L-arabinan and both arabinogalactan type I (AG-I) and II chains linked to rhamnogalacturonan I (RG-I) backbone linked to longer galacturonan chains. In vitro bioactivity study showed that PGP-I-I could restore the intestinal cellular antioxidant defense under the condition of hydrogen peroxide (H2O2) treatment through promoting the expressions of cellular antioxidant genes and protect against oxidative damages.

Topical Exposure to Nemopilema nomurai Venom Triggers Oedematogenic Effects: Enzymatic Contribution and Identification of Venom Metalloproteinase.

Scyphozoan envenomation is featured as severe cutaneous damages due to the toxic effects of venom components released by the stinging nematocysts of a scyphozoan. However, the oedematogenic property and mechanism of scyphozoan venoms remain uninvestigated. Here, we present the oedematogenic properties of the nematocyst venom from Nemopilema nomurai (NnNV), a giant stinging scyphozoan in China, for the first time, using in vivo and in vitro models with class-specific inhibitors. NnNV was able to induce remarkable oedematogenic effects, including induction of significant oedema in the footpad and thigh of mouse, and increase in vascular permeability in the dorsal skin and kidney. Moreover, batimastat, a specific metalloproteinase inhibitor, could significantly reduce the Evan's blue leakage in the damaged organs and attenuate paw oedema after 12 h, but exerted no influence on NnNV-induced thigh oedema. These observations suggested a considerable contribution of NnNV metalloproteinase-like components to the increased vasopermeability, and the participation was strongly suggested to be mediated by destroying the integrity of the vascular basement membrane. Moreover, partial isolation combined LC-MS/MS profiling led to identification of the protein species Nn65 with remarkable metalloproteinase activity. This study contributes to the understanding of the effector components underlying the cutaneous damages induced by scyphozoan stings.

A Novel Digestive α-Amylase from Blue Crab (Portunus segnis) Viscera: Purification, Biochemical Characterization and Application for the Improvement of Antioxidant Potential of Oat Flour.

This study reports on the purification and characterization of a digestive α-amylase from blue crab (Portunussegnis) viscera designated Blue Crab Amylase (BCA). The enzyme was purified to homogeneity by ultrafiltration, Sephadex G-100 gel filtration and Sepharose mono Q anion exchange chromatography, with the final purification fold of 424.02, specific activity of 1390.8 U mg-1 and 27.8% recovery. BCA, showing a molecular weight of approximately 45 kDa, possesses desirable biotechnological features, such as optimal temperature of 50 °C, interesting thermal stability which is enhanced in the presence of starch, high stability towards surfactants (Tween 20, Tween 80 and Triton X-100), high specific activity, quite high storage and broad pH range stability. The enzyme displayed Km and Vmax values, of 7.5 ± 0.25 mg mL-1 and 2000 ± 23 µmol min-1 mg-1 for potato starch, respectively. It hydrolyzed various carbohydrates and produced maltose, maltotriose and maltotetraose as the major end products of starch hydrolysis. In addition, the purified enzyme was successfully utilized for the improvement of the antioxidant potential of oat flour, which could be extended to other cereals. Interestingly, besides its suitability for application in different industrial sectors, especially food industries, the biochemical properties of BCA from the blue crab viscera provide novel features with other marine-derived enzymes and better understanding of the biodegradability of carbohydrates in marine environments, particularly in invasive alien crustaceans.