Miyabenol A inhibits LPS-induced NO production via IKK/IkappaB inactivation in RAW 264.7 macrophages: possible involvement of the p38 and PI3K pathways

The anti-inflammatory effect of miyabenol A, a stilbene isolated from Vitis thunbergii, on lipopolysaccaride (LPS)-induced nitric oxide (NO) production in RAW264.7 macrophages was studied. Miyabenol A inhibited NO production (EC 50: 2.7 muM) and iNOS protein and mRNA expression in a parallel concentration-dependent manner. LPS-evoked NF-kappaB nuclear translocation and associated IkappaB degradation were abrogated by miyabenol A treatment. Phosphorylations of IKKalpha/beta, ERK1/2, JNK p38 MAPK, and Akt were observed in LPS-stimulated cells; nevertheless, miyabenol A selectively blocked IKKalpha/beta, p38, and Akt phosphorylation. Furthermore, LPS-stimulated IKKalpha/beta and Akt phosphorylation was abolished by p38 inhibitor SB203580. Wortmannin (a PI3K inhibitor) also attenuated LPS-induced IKKalpha/beta phosphorylation, although to a less extent than SB203580, but failed to affect p38 phosphorylation. These observations suggested that PI3K/Akt might lie downstream of p38 MAPK to coregulate LPS-induced IKKalpha/beta phosphorylation. Taken together, miyabenol A acted via interfering with p38 MAPK-related signal pathways to down-regulate IKK/IkappaB activation and NO production.     

Chemical composition of Greek avgotaracho prepared from mullet (Mugil cephalus): nutritional and health benefits

Crude composition, lipid composition, and tocopherols, ascorbic acid, cholesterol, phytosterols, and squalene content together with fatty acids and antiplatelet activities of total, neutral, and polar lipids of avgotaracho (wax-covered, dried, and salted Mugil cephalus roe) were studied and compared with those of similar products. Wax and steryl esters accounted for 63.7% of roe lipids followed by phosphatidylcholine (PC), which comprised 20.3%. Wax esters were rich in saturated fatty alcohols, monounsaturated fatty acids, and long chain omega3 highly unsaturated fatty acids (HUFA). The fatty acid distribution in roe total and neutral lipids was similar to that of wax esters, while in polar lipids, the omega3 HUFA predominated. Avgotaracho provides significant amounts of protein, fat, alpha-tocopherol, ascorbic acid, and PC, certain amounts of squalene and phytosterols, and cholesterol at levels comparable to hens' eggs. Total, polar, and neutral lipids of avgotaracho exhibited a strong inhibition of platelet activating factors and thrombin, with polar lipids being more active. The results obtained indicate that avgotaracho is a food of high nutritive value, rich in protein and lipids with a healthy lipid profile in terms of omega3/omega6 ratio and major fatty acid classes, while the antiplatelet activity of its oil indicates a putative antithrombotic potential.     

A modified chemical procedure for rapid determination of glucosamine and its application for estimation of mold growth in peanut kernels and koji.

One-step hydrolysis of chitin to release glucosamine for quantitation was achieved by combining a chitin-containing sample (10-200 mg of sample size) in a test tube with 1 mL of 10 M HCl followed by vacuum treatment for 10 min, incubation at 28 degrees C for 30 min, replenishment with 3 mL of deionized water, nitrogen flushing, screw capping, and heat treatment at 140 degrees C for 60 min. A phosphate buffer solution (pH 12.5, 0.2 M) was effective in pH stabilization and enhancing colorimetric determination of glucosamine content. When the modified procedure was applied to analyze glucosamine content in the mycelia of various molds, glucosamine content varied mainly depending on mold species. In estimations of mold growth of the uninoculated peanut kernels incubated under a humidified condition for 5 weeks, cooked rice and soybean inoculated with conidia of Aspergillus oryzae for koji preparation, logarithms of the internal mold populations and glucosamine contents both increased with increases of incubation time. The modified procedure provided a rapid and reliable estimation of mold growth in various substrates.     

Immunopathological effects of porcine circovirus type 2 (PCV2) on swine alveolar macrophages by in vitro inoculation

Porcine circovirus type 2 (PCV2) is the primary causative agent of postweaning multisystemic wasting syndrome (PMWS), a multifactorial disease, in pigs. Monocyte/macrophage lineage cells, including alveolar macrophages (AMs), are the major target cells for PCV2. Swine AMs are essential for the pulmonary defense system against various pathogens. Concurrent infection of lung with opportunistic pathogens in pigs suffered from PMWS is speculated as a feature of immunosuppression. The present study was conducted to characterize the effects of PCV2 inoculation on swine AMs in the in vitro system. The parameters selected for evaluation included PCV2 antigen- and nucleic acid-containing rate, viability, TUNEL-positive rate, phagocytosis, microbicidal capability, and capacity for production of reactive oxygen species (superoxide anion, O₂⁻, and hydrogen peroxide, H₂O₂), cytokines, and chemokines. High intracytoplasmic PCV2 antigen- and nucleic acid-containing rate, absence of intranuclear signals for PCV2 antigen and nucleic acid, and lack of noticeable cell death were seen in PCV2-inoculated AMs. The PCV2-inoculated AMs displayed a transient as well as persistent reduction in the up-take and destruction of Candida albicans, respectively, accompanied by decrease in the production of O₂⁻ and H₂O₂. In PCV2-inoculated AMs, the levels of tumor necrosis- (TNF-) and interleukin-8 (IL-8) were significantly increased; the mRNA expression levels of alveolar macrophage-derived neutrophil chemotactic factors-II (AMCF-II), granulocyte colony-stimulating factor (G-CSF), monocyte chemotactic protein-1 (MCP-1), and IL-8 were strongly up-regulated. The reduced phagocytosis and microbicidal capability in conjunction with decreased production of reactive oxygen species in PCV2-inoculated AMs suggest that PCV2-containing AMs may favor the survival and spread of PCV2. It is speculated that the functional alterations observed in PCV2-containing AMs may be potentially harmful to the lung tissue and local pulmonary defense system, especially in those PCV2-infected pigs conditioned by various PMWS development-dependent co-factors.     

Differential expression of U2AF35 in the arthritic joint of avian reovirus-infected chicks

To identify cell types and genes that are differentially expressed during immunopathogenesis of avian reovirus (ARV)-induced viral arthritis (VA), we inoculated arthrotropic strain S1133 of ARV into 1-day-old broilers, and examined tissue histology as well as RNA expression at different days post-inoculation (PI). Using immunohistochemical staining, we detected many CD68 expressing macrophages in and around the blood vessels of the arthritic joints. By RT-PCR, we found that expression of matrix metalloproteinase-2 (MMP-2) and bone morphogenetic protein-2 (BMP-2) was induced earlier in footpads and hock joints of ARV-infected chickens. By employing suppression subtractive hybridization (SSH) technique and RT-PCR, we further identified that small subunit of U2 snRNP auxiliary factor (U2AF35 or U2AF1) mRNA was differentially induced in the joint of ARV-infected chickens. By in situ hybridization (ISH), mRNA signals of U2AF35 and BMP-2 were located in chondrocytes within/near the epiphyseal plate and secondary center of ossification, and in epidermal cells and dermal fibroblast-like cells of arthritic joints. In addition, U2AF35 mRNA was expressed in the inflammatory infiltrates of the bone marrow of ARV-infected arthritic joints, while MMP-2 was mainly detected in chondrocytes. Interestingly, among U2AF35, MMP-2, and BMP-2 that were differentially expressed in the joint of ARV-infected chickens, only U2AF35 induction correlated well with arthritic manifestation. Because U2AF35 may assist in mRNA splicing of proinflammatory chemokines and cytokines, our results indicated that U2AF35 induction might play an immunopathological role in ARV-induced arthritis. This study has first associated U2AF35 to viral arthritis.     

Neuroprotective effects of resveratrol on cerebral ischemia-induced neuron loss mediated by free radical scavenging and cerebral blood flow elevation

Resveratrol is a natural phytoestrogen and possesses many biological functions such as anti-inflammatory activity and protection against atherosclerosis and myocardial infraction. The present study was carried out to elucidate the neuroprotective effect and possible mechanism of resveratrol on cerebral ischemia-induced hippocampus neuron loss. Sixty adult male rats underwent general anesthesia (urethane, 1.4 g/kg, i.p.) and were divided into three groups: sham operation, ischemia treatment, and ischemia combined with resveratrol administration (20 mg/kg, i.v.). The carotid artery was bilaterally ligated to induce cerebral ischemia. Microdialysis and high-performance liquid chromatography were used to analyze dihydroxybenzoic acid (DHBA) that reflected the hippocampal hydroxyl radical level. Hippocampal nitric oxide was assayed among different groups. During cerebral ischemia, the hydroxyl radical levels were elevated in rats and animals displayed severe neuronal loss. A single dose of resveratrol significantly increased the nitric oxide level and decreased the hydroxyl radical level. The reduction of cerebral blood flow and neuronal loss were also attenuated by resveratrol treatment. The results demonstrated that a single infusion of resveratrol could elicit neuroprotective effects on cerebral ischemia-induced neuron damage through free radical scavenging and cerebral blood elevation due to NO release.     

Development of probe-based ultraweak chemiluminescence technique for the detection of a panel of four oxygen-derived free radicals and their applications in the assessment of radical-scavenging abilities of extracts and purified compounds from food and herbal preparations

We report here the development of a probe-based ultraweak chemiluminescence (uwCL) method capable of detecting a panel of four oxygen-derived free radicals (ODFRs) including superoxide (O2-), hydrogen peroxide (H2O2), hydroxyl radical (*OH), and peroxyl radical (ROO*) using different probes specific for these radicals performed by the same uwCL analyzer. The selected radical-generating systems and their corresponding uwCL-probing emitters were validated. These ODFR-detecting systems were subsequently utilized by us to assess the radical-scavenging ability (RSA) of a variety of extracts and purified constituents derived from foods and herbal preparations. Our approach for assessing RSA for these constituents is based on the suppression of uwCL generated by each ODFR, and the degrees of inhibition have been shown to be dose-dependent. For this reason, the estimation of IC50 for each testing compound can be obtained from the curve constructed based on the percent of inhibitions of uwCL versus the concentrations of the compound tested. To illustrate the practical applications of our devised methodology, data for comparative studies of RSA activities of fermented extracts of Cordeceps sinensis, purified methylgallate isolated from Toona sinesis, resveratrol purified from grape seeds, plus epimedin C from the aerial part of the Epimedium plant (yinyanghuo) are to be presented.     

Comparative characterization of peanuts grown by aquatic floating cultivation and field cultivation for seed and resveratrol production

Peanut pods (Tainan 12, a Spanish cultivar, Arachis hypogaea L.) have been obtained from peanuts grown in a newly developed aquatic floating cultivation system without artificial aeration or periodic renewal of the solution. The system provided a convenient status for examination of root and pod development. Compared to field-grown peanuts of the same cultivar, the aquatic-cultivated peanut pods and seeds were smaller, whereas seed/pod weight ratios, crude fat and protein contents, and SDS-PAGE protein patterns varied within similar ranges. During cultivation, the highest detected temperature of the aquatic solution was higher than the field-soil temperature. After gas chromatographic analysis of the fatty acid compositions, the oleic acid/linoleic acid ratio of the aquatic-cultivated seeds was higher than that of field-cultivated ones. When the peanut roots were collected, cleaned, dried, weighed, pulverized, and subjected to resveratrol analysis, dry root weights were 4.2 +/- 0.1 and 2.2 +/- 1.1 g/plant and resveratrol contents were 0.074 +/- 0.009 and 0.114 +/- 0.212 mg/g for the aquatic- and field-cultivated peanut roots, respectively. This indicates that the aquatic-cultivated peanut roots could be a potent and consistent source of resveratrol.