Pterostilbene is more potent than resveratrol in preventing azoxymethane (AOM)-induced colon tumorigenesis via activation of the NF-E2-related factor 2 (Nrf2)-mediated antioxidant signaling pathway

Inflammatory bowel diseases have been a risk factor of colorectal cancer (CRC). The reactive oxygen species (ROS) generated by inflammatory cells create oxidative stress and contribute to neoplastic transformation, proliferation, and even metastasis. Previously, resveratrol (RS) and pterostilbene (PS) had been reported to prevent chemical-induced colon carcinogenesis by anti-inflammatory and pro-apoptotic properties. In this study, we investigated whether RS and PS could prevent the azoxymethane (AOM)-induced colon tumorigenesis via antioxidant action and to explore possible molecular mechanisms. Male BALB/c mice were injected with AOM (5 mg/kg of body weight) with or without RS or PS, and at the end of the protocol, all of the mice were euthanized and colons were analyzed. Administrations of PS can be more effective than RS in reducing AOM-induced formation of aberrant crypt foci (ACF), lymphoid nodules (LNs), and tumors. We also find that PS is functioning more effectively than RS to reduce nuclear factor-κB (NF-κB) activation by inhibiting the phosphorylation of protein kinase C-β2 (PKC-β2) and decreasing downstream target gene expression, including inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), and aldose reductase (AR) in mouse colon stimulated by AOM. Moreover, administration of RS and PS for 6 weeks significantly enhanced expression of antioxidant enzymes, such as heme oxygenase-1 (HO-1) and glutathione reductase (GR), via activation of NF-E2-related factor 2 (Nrf2) signaling. When the above findings are taken together, they suggest that both stilbenes block cellular inflammation and oxidative stress through induction of HO-1 and GR, thereby preventing AOM-induced colon carcinogenesis. In comparison, PS was a more potent chemopreventive agent than RS for the prevention of colon cancer. This is also the first study to demonstrate that PS is a Nrf2 inducer and AR inhibitor in the AOM-treated colon carcinogenesis model.     

Apparatus used for small-scale volatile extraction from ethanol-supplemented low-salt miso and GC-MS characterization of the extracted flavors

An extraction apparatus was equipped with a nitrogen-flushing vessel to purge volatiles from a 10-g miso prepared solution at 40 degrees C, a reflux condenser to recover water, a coiled cold-trap to separate ethanol in advance, and a glass-lined stainless (GLS) trap filled with Tenax TA for flavor adsorption. Volatiles in the GLS tube were released with a thermal desorption device and condensed with a Micro-cryo trap prior to connection with GC and GC-MS for characterization. After analysis, a broad volatile profile comprising 9 categories of functional group and 97 identified compounds was achieved. As affected by ethanol supplementation for miso fermentation, most volatiles except alcohols and acetals in the low-salt products fermented with 5% NaCl and 7.5% ethanol were higher than those in the control products fermented with 9% NaCl and 0% ethanol and the high-ethanol supplemented products fermented with 5% NaCl and 15% ethanol. It reveals that supplementation of ethanol in miso at an appropriate level not only enabled a low-salt miso fermentation but also enhanced flavor formation.     

Squalene content and antioxidant activity of Terminalia catappa leaves and seeds

Squalene was identified by gas chromatography-mass spectrometry and high-performance liquid chromatography (HPLC) spiking analyses in the supercritical CO(2) extracts of freeze-dried abscisic leaves of Terminalia catappa L. When the freeze-dried abscisic, senescent, mature, and immature leaves and seeds were subjected to supercritical CO(2) extraction at 40 degrees C and 3000 psi and HPLC quantitation, squalene contents were 12.29, 2.42, 1.75, 0.9, and 0% in the extracts and corresponding to 1499, 451, 210, 65, and 0 microg/g in the freeze-dried sample, respectively. When the extracts were applied for antioxidative characterization by supplementation in an iron/ascorbate system with linoleic acid and in a pork fat storage system for inhibition of conjugated diene hydroperoxide (CDHP) formation or in a free radical scavenging system with 1,1-diphenyl-2-picryl-hydrazyl (DPPH), the extracts of leaves exhibited potent antioxidative and DPPH scavenging activities and increased with an increase of leaf maturity. However, the seed extracts only exhibited potent inhibition of CDHP formation and very low DPPH scavenging activity.     

Seroepidemiologic survey of Dirofilaria immitis infection among domestic dogs in Taipei City and mountain aboriginal districts in Taiwan (1998-1999).

To estimate the seroprevalence of Dirofilaria immitis infection in domestic dogs in Taiwan, we utilized a commercial ELISA kit (Snap, IDEXX, USA) for detecting circulating antigens released by adult female worms. Serum specimens of 664 domestic dogs sampled from Taipei City in northern Taiwan and 14 mountain aboriginal districts in eastern Taiwan were screened for D. immitis antigens. Multivariate-adjusted odds ratios (ORs) with their 95% confidence intervals (CIs) were estimated by multiple logistic regression analysis. A total of 89 subjects were antigen-positive, giving a seroprevalence of 13.4%, of which the seroprevalence in Taipei City and mountain aboriginal districts were 13.8 and 12.1%, respectively. The mean overall seropositive rates were 6.3% in 1-3-year-old age group, 14.1% in 3-6-year-old age group and 23.7% in the > or =6-year-old age group. The older the age, the higher the seroprevalence (OR=4.6, 95% CI=2.4-9.0 for the > or =6-year-old age group versus 1-3-year-old age group, P<0.001) for all the dogs in the present study. Moreover, seroprevalence was not different between female and male dogs in either Taipei City or mountain aboriginal districts (P>0.05). Also, no significant difference in seroprevalence among dogs between the two geographical areas was found (P>0.05). In the logistic regression analysis, the seroprevalence of D. immitis remained significantly increased with age after multivariate adjustment in the present study. To our knowledge, this is the first report on the status of D. immitis infection in domestic dogs in Taipei City and mountain aboriginal districts in Taiwan to date.     

Use of a recombinant protein containing major epitopes of hnRNP G to detect anti-hnRNP G antibodies in dogs with systemic lupus erythematosus

The objective of this study was to express major epitopes of heterogeneous nuclear ribonucleoprotein G (hnRNP G) for detecting anti-hnRNP G antibodies in dogs with systemic lupus erythematosus (SLE). HnRNP G cDNA clone was isolated from HEp-2 cells, and a DNA fragment encoding immunodominant region (residues 189-272) of hnRNP G (hnRNP Gi) was subcloned into pET32 vector to construct a prokaryotic expression plasmid named pEThnRNPGi. After induction, Escherichia coli carrying pEThnRNPGi expressed a recombinant protein of 28 kDa, comprising recombinant hnRNP Gi and fusion tag. Purified recombinant hnRNP Gi protein was further analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) and its identity was confirmed. Western blot analysis showed that recombinant hnRNP Gi was specifically recognized by anti-hnRNP G positive sera of SLE dogs, and not by negative control sera. In conclusion, recombinant hnRNP Gi protein expressed in this study may serve as a useful reagent to assist in the immunological diagnosis of canine SLE.     

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.     

Improved superoxide-generating system suitable for the assessment of the superoxide-scavenging ability of aqueous extracts of food constituents using ultraweak chemiluminescence

In the interest of developing a simple and rapid ultraweak chemiluminescence assay for assessing the superoxide (O(2)(-))-scavenging activities of various aqueous extracts of food constituents, a specific and stable O(2)(-)-generating system was sought. Reported herein is the obtainment for the first time of a specific and stable O(2)(-)-generating system consisting of methylglyoxal (MG), a reactive 2-oxo aldehyde and arginine, which has been shown to produce much steadier lucigenin-based chemiluminesence (LBCL) than the conventional xanthine/xanthine oxidase system running in parallel and monitoring by an ultraweak chemiluminescence analyzer. Upon mixing of MG and arginine in a phosphate-buffered saline solution, pH 7.4, steady, time-dependent increments of LBCL can be visually observed. The plateau of LBCL can be reached in approximately 10 min and retained in a steadily stable state thereafter without fluctuation for the next 15 min. The lucigenin-based LBCL generation was shown to be specific since it could be effectively inhibited by active bovine SOD, but not by heat-inactivated enzyme or catalase. Conversely, the xanthine/xanthine oxidase system can merely produce a LBCL peak rapidly but decay instantaneously. To illustrate the application of the proposed method for assessing the O(2)(-)-scavenging ability of various food extracts, namely, Prunus mume (A), Lilum lancifolium (B), Creataegus pinnatifida (C), Tremella fuciformis (D), Fortunella margarita (E), and Scutellaria baicalensis (F), we used the following protocol: 12 min after monitoring of LBCL, 1 mg/mL of each of the test compounds was added to the assay system and various degrees of sudden drop of LBCL values were observed, indicating differences in O(2)(-)-scavenging abilities exerted by these food extracts that can be visually compared. Consequently, the percentages of inhibition of LBCL versus the concentrations of a test compound can be constructed. It follows that the concentration needed to inhibit 50% of LBCL (IC(50)) of a test compound can be extrapolated from the curve. Using this approach, we were able to obtain the IC(50) values of various compounds to be tested and the order of inhibitory efficiency of the above-mentioned food extracts was ranked, being A > B > C > D > E > F, respectively.     

Genetic variation of the lambdaA and lambdaC protein encoding genes of avian reoviruses

Sequence and phylogenetic analysis of lambdaA and lambdaC protein encoding genes of 12 avian reoviruses is described. The sequence of lambdaA possesses a variable region (residues 19-51) located within a conserved hydrophilic region (residues 1-110) and a C(2)H(2) zinc-binding motif (residues 182-202). lambdaC shows the two conserved K residues at positions 169 and 188 indicative of guanylyltransferase activity, an ATP/GTP-binding site motif A (residues 379-386), and a conserved S-adenosyl-l-methionine-binding motif (residues 822-830). Pairwise sequence comparisons show that the mean sequence identities of lambdaA encoding genes and lambdaA proteins are 92% and 98%, respectively, and those of lambdaC encoding genes and lambdaC proteins are 91% and 95%, respectively. Phylogenetic analysis of lambdaA and lambdaC encoding genes reveals that both encoding genes have diverged into three distinct lineages, respectively, and that there is no correlation between lineages and viral serotypes or pathotypes. Also, reassortment of gene segments L1 and L3 has been observed between viruses.     

Suppurative meningitis in a 7-day-old Formosan sambar deer (Cervus unicolor swinhoei) caused by Escherichia coli

This article describes the clinical and pathological features of an orphan 7-day-old, male Formosan sambar fawn that was hospitalized for treatment of weakness. The fawn had been deprived of colostrum and developed suppurative meningitis that was attributed to Escherichia coli.