Isolation of Araguspongine M,a New Stereoisomer of an Araguspongine/Xestospongin alkaloid,and Dopamine from the Marine Sponge Neopetrosia exigua Collected in Palau

A new stereoisomer of an araguspongine/xestospongin alkaloid, named araguspongine M (1), has been isolated together with 12 known compounds, araguspongines B (2) and D (3), dopamine, three galactosyl diacylglycerols, 24-methyl cholesterol, 5,6-dihydrocholesterol, β-sitosterol, and three 5α,8α-epidioxy sterols (11–13), from the marine sponge Neopetrosia exigua (formerly Xestospongia exigua) collected in Palau. The structure of 1 was assigned on the basis of its spectral data analysis. This is the first report on the isolation of dopamine from a marine sponge. This compound may be produced by an endosymbiotic Synechococcus-like cyanobacterium. Compounds 1–3 and 11–13 showed cytotoxicity against HL-60 at IC₅₀’s of 5.5, 5.5, 5.9, 22.4, 9.5, and 9.6 μM, respectively. The possible biosynthesis origin of the isolated metabolites is discussed.     

Antibacterial Bilayered Skin Patches Made of HPMA and Quaternary Poly(4-vinyl pyridine)

This study aimed to produce poly(4-vinyl pyridine) and hydroxypropyl methacrylamide (HPMA)-based bilayer wound dressings materials enhancing healing mechanism for the wounds which have self-healing problem and high infection risk. These materials were designed to protect wound from secondary traumas caused microorganism invasion and do not have toxic substance release problem. Synthesis of quaternary poly(4-vinyl pyridine) (poly(Q4-VP)) which is the antibacterial layer of wound dressing material was carried out in two stages. At first stage, poly(4-vinyl pyridine) polymer was synthesized from 4-vinyl pyridine monomer by free radical polymerization. Then, poly(Q4-VP) was synthesized from poly(4-VP) by alkylation reaction with 6-bromocaproic acid. Resulted polymer was structurally characterized by FT-IR. The macroporous spongy structure, as the lower layer of wound dressing material, was prepared by cryogelation of HPMA. Then, the antibacterial polymer was electrospun onto the cryogel structure and bilayered material was obtained. Cryogel structure, fiber morphology and layer integration was examined by SEM. In order to enhance wound healing process, ascorbic acid (vitamin C) was loaded to cryogel layer and release was followed by spectrophotometrically. The antimicrobial properties of the materials were examined against Escherichia coli, Staphylococcus aureus and Candida albicans, respectively. According to the results, bilayered, antibacterial and antifungal against Staphylococcus aureus and Candida albicans, temporary wound dressings which can stimulate wound healing and have high swelling capacity were obtained successfully.     

Reduction of ochratoxin a levels in red wine by bentonite, modified bentonites, and chitosan

Adsorption method may play an important role to remove ochratoxin A (OTA) from wine by bentonite (B), nonylammonium bentonite (NB), dodecylammonium bentonite (DB), KSF-montmorillonite (KSF), and chitosan bead (CB). The optimum conditions of OTA adsorption from synthetic solutions were revealed at room temperature and pH 3.5. The adsorption equilibria of B and NB were almost established within 120 and 240 min, respectively. DB, KSF, and CB had about 90 min of equilibration time. The adsorption efficiency carried out in the synthetic OTA solution did not change remarkably when the amounts of adsorbents were 25 mg for bentonite, DB, and KSF and 100 mg for NB and CB. Furthermore, 25 mg of adsorbents was used at all adsorption studies in synthetic solution. The adsorption isotherm was fitted with mostly a Freundlich equation with respect to the correlation coefficients. The adsorption data were evaluated using Langmuir and Freundlich equations having Kf values ranging from 0.011 to 9.5 with respect to correlation coefficients (R2 = 0.900-0.977). DB and KSF have the highest adsorption capacity for OTA in synthetic solutions. In wine, the removal of OTA was succeeded at a percentage of 60-100 by KSF and CB. Furthermore, the highest adsorption capacity of OTA for red wine was obtained by using 250 mg of KSF, which caused less damage to the nature of wine and also low adsorption of polyphenols and anthcyans.     

Antioxidative stress activity of oligophosphopeptides derived from hen egg yolk phosvitin in Caco-2 cells

The protective effects of hen egg yolk phosvitin phosphopeptides (PPPs) against hydrogen peroxide (H2O2)-induced oxidative stress were evaluated in an in vitro assay using human intestinal epithelial cells. Caco-2 cells were stimulated with 1 mM H2O2 for 6 h, and the secretion of IL-8, a proinflammatory mediator, was determined by ELISA as a biomarker of oxidative stress. The inhibition of H2O2-induced IL-8 secretion from Caco-2 cells was observed by pretreatment for 2 h with PPPs, but not with phosvitin. PPPs also suppressed the formation of malondialdehyde in H2O2-treated Caco-2 cells. Furthermore, intracellular glutathione levels and glutathione reductase activity were elevated by the addition of PPPs. The protective effects of PPPs against H2O2-induced oxidative stress were almost the same as that of glutathione, and PPPs with a high content of phosphorus exhibited higher protective activity than PPPs without phosphorus; however, phosphoserine itself did not show any significant antioxidative stress activity. These findings suggest that oligophosphopeptides from hen egg yolk phosvitin possess novel antioxidative activity against oxidative stress in intestinal epithelial cells and that phosphorus and peptide structure seem to have a key role in the activity.     

Oligophosphopeptides derived from egg yolk phosvitin up-regulate gamma-glutamylcysteine synthetase and antioxidant enzymes against oxidative stress in Caco-2 cells

Previously, we have found phosphopeptides (PPPs) from hen egg yolk phosvitin possess a potent antioxidative activity against oxidative stress in human intestinal epithelial cells, Caco-2. However, their biological activity at the cellular level has not yet fully understood. The objective of this study is to evaluate the regulation of glutathione (GSH) biosynthesis-associated and antioxidant enzymes against oxidative stress in Caco-2 cells using an in vitro model. Treatment of 1 mM H2O2-induced Caco-2 cells with PPPs increased cellular GSH levels, concomitant with a significant increase in gamma-glutamylcysteine synthetase (gamma-GCS) activity and the expression of gamma-GCS heavy subunit mRNA. Furthermore, intracellular glutathione reductase, glutathione S-transferase, and catalase activities were elevated by PPPs. In addition, PPPs with high content of phosphorus showed higher induction of these enzyme activities than PPPs without phosphorus. These data indicate that oligophosphopeptides from hen egg yolk phosvitin can up-regulate cellular GSH biosynthesis-associated enzymes activity and antioxidative activities, which play key roles against tissue oxidative stress in the human intestinal epithelial cells.     

Arylamidase and amidohydrolases in soils as affected by liming and tillage systems

This study investigated the long-term effect of lime application and tillage systems (no-till, ridge-till and chisel plow) on the activities of arylamidase and amidohydrolases involved in N cycling in soils at four long-term research sites in Iowa, USA. The activities of the following enzymes were assayed: arylamidase, -asparaginase, -glutaminase, amidase, urease, and -aspartase at their optimal pH values. The activities of the enzymes were significantly (P<0.001) and positively correlated with soil pH, with r values ranging from 0.42^* to 0.99^*** for arylamidase, 0.81^*** to 0.97^*** for -asparaginase, 0.62^*** to 0.97^*** for -glutaminase, 0.61^*** to 0.98^*** for amidase, 0.66^** to 0.96^*** for urease, and 0.80^*** to 0.99^*** for -aspartase. The Δactivity/ΔpH values were calculated to assess the sensitivity of the enzymes to changes in soil pH. The order of the sensitivity of enzymes was as follows: -

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-aspartase. The enzyme activities were greater in the samples of the 0–5 cm depth than those of the 0–15 cm samples under no-till treatment. Most of the enzyme activities were significantly (P<0.001) and positively correlated with microbial biomass C (C_mic) and N (N_mic). Lime application significantly affected the specific activities of the six enzymes studied. Results showed that soil management practices, including liming and type of tillage significantly affect soil biological and biochemical properties, which may lead to changes in nitrogen cycling, including N mineralization in soils.     

Simultaneous liquid chromatographic determination of residual synthetic antibacterials in cultured fish

A liquid chromatographic method is described to determine simultaneously the following 11 synthetic antibacterial agents used in a fishery: nitrofuran derivatives furazolidone, nifurpirinol, difurazone, and furamizole; sulfa drugs sulfamerazine, sulfisozole, sulfamonomethoxine, and sulfadimethoxine; and, oxolinic, nalidixic, and piromidic acids. A Nucleosil C18 column was used with tetrahydrofuran-acetonitrile-phosphoric acid-water (29 + 1 + 0.06 + 69.94) as the mobile phase. Pretreatment of the fish meat sample with acetone extraction and alumina column cleanup gave good separation of the LC peaks without interference from any other components. Recovery of the antibacterial agents was ca 80%. The lower limit of detection of the drugs was 1-2 ng for 10 microL injection.     

Study of thermal aggregation of oat globulin by laser light scattering

The heat-induced aggregation of oat globulin was studied using size-exclusion chromatography (SEC) combined with on-line multiangle laser light scattering (MALLS) and quasi-elastic light scattering (QELS). The unheated oat globulin exists as a mixture of hexamer (>95%), trimer, and dimer forms of hexamer. The molecular weight of the hexamer was estimated by MALLS to be 330 000, close to that deduced from the genomic cloned data of the acidic and basic polypeptides of oat globulin. From QELS measurements, it can be predicted that the hexamer exists as two annular trimeric rings, with a diameter of 11.8 nm, placed on top of each other, forming an oblate cylinder with a height of about 8.5 nm. Upon heating at 100 degrees C, the oat globulin hexamers and trimers were dissociated into monomers. The heat-denatured monomers, probably assuming an extended structure, were associated to form small aggregates, which were further aggregated to high molecular weight complexes. Upon further heating (60 min), the soluble aggregates were associated to form insoluble aggregates. Aggregation of oat globulin occurred at a much faster rate at 110 degrees C. The results indicate that the SEC-MALLS-QELS system is suitable for studying thermal aggregation of food proteins.     

Chicken eggshell matrix proteins enhance calcium transport in the human intestinal epithelial cells, Caco-2

Chicken eggshell powder has been proposed as an attractive source of calcium for human health to increase bone mineral density in an elderly population with osteoporosis. However, factors affecting calcium transport of eggshell calcium have not yet been evaluated. Chicken eggshell contains about 1.0% (w/w) matrix proteins in addition to a major form of calcium carbonate (95%, w/w). In this study, we found that soluble eggshell matrix proteins remarkably enhance calcium transport using in vitro Caco-2 cell monolayers grown on a permeable support. The total calcium transport across Caco-2 monolayers showed an increase of 64% in the presence of 100 microg/well soluble eggshell matrix proteins. The active enhancer with a molecular mass of 21 kDa was isolated by reversed phase high-performance liquid chromatography and did not correspond to any previously identified protein. The N-terminal sequence was determined to be Met-Ala-Val-Pro-Gln-Thr-Met-Val-Gln. The possible mechanisms of eggshell matrix protein-mediated increase in calcium transport and the potential significance of eggshell calcium as a nutraceutical are discussed.