Effects of fermented red bean extract on nephropathy in streptozocin-induced diabetic rats.

BACKGROUND: The antioxidant effects of Bacillus subtilis-fermented red bean (natto-red bean) extract (NRBE) in young (6 weeks old) Sprague-Dawley rats and aged (12 months old) mice had been reported previously. OBJECTIVE: To evaluate the antioxidant and anti-inflammatory effects of NRBE in the kidneys of streptozocin-induced diabetic rats. DESIGN: Normal control rats and diabetic rats were orally gavaged with saline and low-dose NRBE (100 mg/kg body weight [BW]), medium-dose NRBE (200 mg/kg BW), and high-dose NRBE (500 mg/kg BW), for 12 weeks and then sacrificed. Concentration of fasting glucose, adiponectin, renal function markers, antioxidative markers, and pro-inflammatory markers were measured. RESULTS: Oral administration of 50% ethanolic extract of NRBE with a dosage of 100 mg/kg BW, 200 mg/kg BW, or 500 mg/kg BW could improve the symptoms of kidney enlargement and renal function. Supplementation of NRBE can effectively inhibit the formation of renal reactive oxygen species and advanced-glycation end-products and increase renal glutathione content and serum adiponectin. A low dose of NRBE (100 mg/kg BW) decreased fasting blood sugar and renal interleukin (IL)-6 expression. Serum C-reactive protein, renal tumor necrosis factor-α, and monocyte chemoattractant protein-1 concentrations were decreased, and renal superoxide dismutase activity was increased in the medium-dose NRBE group. Twenty-four hour creatinine clearance and urinary albumin excretion also improved by medium-dose NRBE supplementation. In NRBE, total phenols and flavonoids were 6.3 mg gallic acid equivalent/g and 12.02 mg rutin equivalent/g, respectively, and kampherol was the major active antioxidant compound. CONCLUSION: This study demonstrated that appropriate amount of NRBE, 200 mg/kg BW in rats, could prevent diabetic nephropathy by improving antioxidant status and inhibiting inflammation in renal tissue.

Locust bean gum galactomannan hydrolyzed by thermostable ß-d-mannanase may reduce the secretion of pro-inflammatory factors and the release of granule constituents.

Locust bean gum (LBG) galactomannan has been claimed to have applications in the biopharmaceutical field. However, the effects of LBG galactomannan on immunomodulatory aspects are not yet clear. The purpose of this study was to over-express thermostable ß-d-mannanase from the thermophilic actinomycete Thermobifida fusca BCRC 19214 using a Pichia pastoris expression system. The maximum intracellular ß-d-mannanase activity obtained from the cell-free extract was approximately 40.0U/mL after 72h of cultivating a P. pastoris transformant (pPICZ-man) induced with methanol. Hydrolysis of native LBG galactomannan with 8U/mL ß-d-mannanase for 24h significantly decreased the weight-average molecular weight of LBG galactomannan from 5,580,010 to 3188. Native and hydrolyzed LBG galactomannan in a range of 0-0.2% did not trigger significant cytotoxicity after 24h of treatment compared with the control. The native LBG galactomannan stimulated RAW 264.7 cells to produce cytokine TNF-α dose-dependently, but there was no significant IL-1ß or nitric oxide production. The native LBG galactomannan also stimulated ß-hexosaminidase secretion in RBL-2H3 cells. After the native LBG galactomannan was hydrolyzed with ß-d-mannanase, all of the immunological properties disappeared. These results suggest the possible immunomodulatory effects of native LBG galactomannan.

Chronic fluoxetine treatment enhances sympathetic activities associated with abnormality of baroreflex function in conscious normal rats.

Data regarding the effects of selective serotonin reuptake inhibitors (SSRIs), which are a common type of antidepressants, on cardiovascular autonomic function are inconsistent. The present study was conducted to determine the effects of chronic fluoxetine, an SSRI, on blood pressure, cardiac autonomic nervous activities and baroreflex control of heart rate. Male Sprague-Dawley rats were treated with fluoxetine (10mg/kg day, p.o.) or saline for 14 weeks. Baroreflex function was determined by the sigmoid logistic method based on the heart rate responses to changes in blood pressure elicited by phenylephrine or sodium nitroprusside infusions. Cardiac sympathetic and parasympathetic tones were determined after methylatropine and propranolol treatments. Vascular responsiveness to acetylcholine, phenylephrine and sodium nitroprusside, and cardiac responsiveness to isoproterenol were determined after ganglionic blockade. Chronic fluoxetine treatment increased plasma levels of adrenaline and noradrenaline, but not nitric oxide. Elevation of blood pressure and heart rate by chronic fluoxetine was accompanied by baroreflex resetting and depressed baroreflex sensitivity. Elevated heart rate was mediated by enhanced sympathetic and depressed parasympathetic tones. The lowered baroreflex sensitivity might be attributed to attenuation of the parasympathetic component of baroreflex function. Chronic fluoxetine also diminished cardiac and vascular responsiveness to isoproterenol and acetylcholine, respectively. The plasma levels of adrenaline and noradrenaline were highly correlated with blood pressure, heart rate and baroreflex sensitivity. In conclusion, our results demonstrate that chronic fluoxetine treatment in normal rats induced predominant sympathoexcitation and depressed parasympathetic activity leading to mild hypertension, tachycardia, and impairment of baroreflex function.

Anti-oxidant activity and major chemical component analyses of twenty-six commercially available essential oils.

This study analyzed 26 commercially available essential oils and their major chemical components to determine their antioxidant activity levels by measuring their total phenolic content (TPC), reducing power (RP), ß-carotene bleaching (BCB) activity, trolox equivalent antioxidant capacity (TEAC), and 1,1-diphenyl-2-picrylhydrazyl free radical scavenging (DFRS) ability. The clove bud and thyme borneol essential oils had the highest RP, BCB activity levels, and TPC values among the 26 commercial essential oils. Furthermore, of the 26 essential oils, the clove bud and ylang ylang complete essential oils had the highest TEAC values, and the clove bud and jasmine absolute essential oils had the highest DFRS ability. At a concentration of 2.5 mg/mL, the clove bud and thyme borneol essential oils had RP and BCB activity levels of 94.56% ± 0.06% and 24.64% ± 0.03% and 94.58% ± 0.01% and 89.33% ± 0.09%, respectively. At a concentration of 1 mg/mL, the clove bud and thyme borneol essential oils showed TPC values of 220.00 ± 0.01 and 69.05 ± 0.01 mg/g relative to gallic acid equivalents, respectively, and the clove bud and ylang ylang complete essential oils had TEAC values of 809.00 ± 0.01 and 432.33 ± 0.01 µM, respectively. The clove bud and jasmine absolute essential oils showed DFRS abilities of 94.13% ± 0.01% and 78.62% ± 0.01%, respectively. Phenolic compounds of the clove bud, thyme borneol and jasmine absolute essential oils were eugenol (76.08%), thymol (14.36%) and carvacrol (12.33%), and eugenol (0.87%), respectively. The phenolic compounds in essential oils were positively correlated with the RP, BCB activity, TPC, TEAC, and DFRS ability.

The Topoisomerase 1 Inhibitor Austrobailignan-1 Isolated from Koelreuteria henryi Induces a G2/M-Phase Arrest and Cell Death Independently of p53 in Non-Small Cell Lung Cancer Cells.

Koelreuteria henryi Dummer, an endemic plant of Taiwan, has been used as a folk medicine for the treatment of hepatitis, enteritis, cough, pharyngitis, allergy, hypertension, hyperlipidemia, and cancer. Austrobailignan-1, a natural lignan derivative isolated from Koelreuteria henryi Dummer, has anti-oxidative and anti-cancer properties. However, the effects of austrobailignan-1 on human cancer cells have not been studied yet. Here, we showed that austrobailignan-1 inhibited cell growth of human non-small cell lung cancer A549 and H1299 cell lines in both dose- and time-dependent manners, the IC50 value (48 h) of austrobailignan-1 were 41 and 22 nM, respectively. Data from flow cytometric analysis indicated that treatment with austrobailignan-1 for 24 h retarded the cell cycle at the G2/M phase. The molecular event of austrobailignan-1-mediated G2/M phase arrest was associated with the increase of p21Waf1/Cip1 and p27Kip1 expression, and decrease of Cdc25C expression. Moreover, treatment with 100 nM austrobailignan-1 for 48 h resulted in a pronounced release of cytochrome c followed by the activation of caspase-2, -3, and -9, and consequently induced apoptosis. These events were accompanied by the increase of PUMA and Bax, and the decrease of Mcl-1 and Bcl-2. Furthermore, our study also showed that austrobailignan-1 was a topoisomerase 1 inhibitor, as evidenced by a relaxation assay and induction of a DNA damage response signaling pathway, including ATM, and Chk1, Chk2, γH2AX phosphorylated activation. Overall, our results suggest that austrobailignan-1 is a novel DNA damaging agent and displays a topoisomerase I inhibitory activity, causes DNA strand breaks, and consequently induces DNA damage response signaling for cell cycle G2/M arrest and apoptosis in a p53 independent manner.

A new pyrene-based Schiff-base: a selective colorimetric and fluorescent chemosensor for detection of Cu(II) and Fe(III).

A new receptor 1 was prepared, for the detection of Cu2+ and Fe3+ in solutions as a colorimetric and fluorescent sensor, respectively. Receptor 1 shows highly selective and sensitive recognition toward Cu2+ and Fe3+ by naked eye UV-Vis and fluorescent color changes in aqueous solution (DMSO/H2O=8/2, v/v), respectively. The sensitivity toward Cu2+ or Fe3+ was not interfered with by the presence of other metal ions such as Mg2+, Cd2+, Ag+, Zn2+, Ni2+, Co2+, Mn2+, Cr3+, Ca2+, Na+, Pb2+, K+, Fe2+, Li+ and Hg2+ ions. Receptor 1 can be used for semi-quantitative recognition of Cu2+ ions at ppm level. The fluorescence microscopy experiments showed that the receptor is efficient for detection of Fe3+ in vitro, developing a good image of the biological organelles.

A new selective colorimetric and fluorescent chemodosimeter for HSO4(-) based on hydrolysis of Schiff base.

Two new receptors 1 and 2 were prepared, and their chromogenic and fluorogenic behaviors toward various anions were investigated. Receptors 1 and 2 show exclusive response toward HSO4(-) ion and also distinguish HSO4(-) from other anions by different color changes in aqueous solution (CH3CN/H2O=4/1, v/v). Between them receptor 1 selectively exhibits a pronounced HSO4(-)-induced fluorescence enhancement. The detection limit for the HSO4(-) ion was determined as (0.24±0.03µM). Thus, the receptor 1 can be used as a colorimetric and fluorescent sensor for the determination of HSO4(-) ion. The sensing mechanism has been suggested to proceed via a hydrolysis process. The hydrolysis product has been isolated and further identified by (1)H NMR spectroscopy, ESI-MS analysis and X-ray diffraction.

Mass spectra of nitro-beta,beta-dihalostyrenes.

The electron ionization (EI) fragmentation of nitro-beta,beta-dihalostyrenes depends strongly on the specific halogens present as well as on the position of the nitro group. Beta,beta-difluorostyrenes yielded mainly fluoroacetylene ions as the base peaks and o-nitro-beta,beta-dihalostyrene possibly furnished the ion with a structure similar to that of benzo[a]isoxazole (11). The structural identity of the ion was verified by comparing the collision-induced dissociation (CID) spectra of m/z 119 ions from p- and o-nitro-beta,beta-difluorostyrene, o-nitro-beta,beta-dibromostyrene and compound 11.