Effect of vitamin D supplementation on pegylated interferon/ribavirin therapy for chronic hepatitis C genotype 1b: a randomized controlled trial.

Chronic HCV-infected patients tend to have vitamin D deficiency, suggesting that vitamin D supplementation may enhance the efficacy of treatment with pegylated interferon (PEG-IFN) and ribavirin (RBV). We therefore assessed the effects of vitamin D supplementation on viral response to PEG-IFN/RBV. Eighty-four patients with HCV genotype 1b were randomized, 42 to oral vitamin D supplementation (1000 IU/day) and 42 to nonsupplementation (control), from week 8 to the end of PEG-IFN/RBV therapy. The primary end point was undetectable HCV RNA at week 24 (viral response [VR]). VR rate at week 24 was significantly higher in the vitamin D than in the control group (78.6% vs 54.8% P = 0.037). Adverse events were similar in both groups. When patients were subdivided by IL28B SNP rs8099917 genotype, those with the TT genotype group showed a significantly higher VR rate at week 24 with than without vitamin D supplementation (86.2% vs 63.3% vs P = 0.044). Although patients with the TG/GG genotype, who were relatively resistant to PEG-IFN treatment, had similar VR rates at week 24 with and without vitamin D supplementation, the decline in viral load from week 8 to week 24 was significantly greater with than without vitamin D supplementation. Multivariate analysis showed that rs8099917 genotype and vitamin D supplementation contributed significantly to VR at week 24. SVR rates were similar in the vitamin D and control groups [64.3% (27/42) vs 50% (21/42), P = 0.19]. Vitamin D supplementation may enhance the effects of PEG-IFN/RBV in HCV genotype 1b-infected patients.

Low-molecular-weight lignin-rich fraction in the extract of cultured Lentinula edodes mycelia attenuates carbon tetrachloride-induced toxicity in primary cultures of rat hepatocytes.

The extract of cultured Lentinula edodes mycelia (LEM) is a medicinal food ingredient that has hepatoprotective effects. In this study, we fractionated the LEM extract to explore novel active compounds related to hepatoprotection by using primary cultures of rat hepatocytes exposed to carbon tetrachloride (CCl(4)). The LEM extract and the fractions markedly inhibited the release of alanine aminotransferase (ALT) from hepatocytes damaged by CCl(4) into the culture medium. The strongest hepatocyte-protective activity was seen in a fraction (Fr. 2) in which a 50% ethanol extract was further eluted with 50% methanol and separated using reverse-phase HPLC. Fr. 2 had an average molecular weight of 2753, and the main components are lignin (49%) and saccharides (36%, of which xylose comprises 41%). Therefore, Fr. 2 was presumed to be a low-molecular-weight compound consisting mainly of lignin and xylan-like polysaccharides. The hepatocyte-protective activity was observed even after digestion of xylan-like polysaccharides in Fr.2 and confirmed with low-molecular-weight lignin (LM-lignin) alone. In addition, Fr. 2, the xylan-digested Fr. 2 and LM-lignin showed higher superoxide dismutase (SOD)-like activity than the LEM extract. These results suggested that the effective fraction in the LEM extract related to hepatocyte protection consisted mainly of LM-lignin, and its antioxidant activity partially contributes to the hepatocyte-protective activity of the LEM extract.

Phylogenetic relationships among wild and cultivated Tartary buckwheat (Fagopyrum tataricum Gaert.) populations revealed by AFLP analyses.

Phylogenetic relationships among cultivated landraces and natural populations of wild subspecies of Tartary buckwheat were investigated at the individual level by constructing a phylogenetic tree based on amplified fragment length polymorphism (AFLP) markers. Seven individuals from seven cultivated landraces and 35 individuals from 21 natural populations of wild subspecies were utilized for AFLP analyses. Three groups were recognized: (1) all cultivated landraces and wild subspecies from northern Pakistan, central and eastern Tibet, and northwestern Yunnan, (2) wild subspecies from central and southern Sichuan, (3) wild subspecies from northern Sichuan and eastern Tibet. It was concluded that cultivated Tartary buckwheat probably originated in eastern Tibet or northwestern Yunnan in China.

Comparative study on reduction of bone loss and lipid metabolism abnormality in ovariectomized rats by soy isoflavones, daidzin, genistin, and glycitin.

The effects of the soy isoflavone glycoside, daidzin, genistin, and glycitin on bone loss and lipid metabolism in ovariectomized (ovx) rats were compared with those of estrone. Thirty-six 11-week-old female Sprague-Dawley rats were assigned to six groups, sham-operated, ovx, ovx+glycitin, ovx+daidzin, ovx+genistin, and ovx+estrone and fed matched amounts of a commercial calcium-deficient diet for 4 weeks. Throughout this period, daidzin, genistin or glycitin (25, 50 or 100 mg/kg/d) was given orally using a stomach tube, or estrone (7.5 microg/kg/d) was administered subcutaneously. Daidzin, genistin and glycitin significantly prevented bone loss in ovx rats at a dose of 50 mg/kg/d, like estrone. At this dose glycitin and daidzin also prevented ovx-induced uterine atrophy and increases in body weight gain, abdominal fat, serum total cholesterol and triglyceride, and urinary excretion of pyridinoline and deoxypyridinoline with statistical significance, like estrone. On the other hand, genistin prevented ovx-induced uterine atrophy only at a dose of 100 mg/kg, but did not block any other change of ovx rats at a dose of 50 or 100 mg/kg. These findings indicate that daidzin, glycitin, and genistin are effective in preventing bone loss and the former two compounds are effective in reversing the unfavorable changes of lipid metabolism in this model. It is suggested that the preventive effect of daidzin or glycitin on bone loss in ovx rats is due to suppression of bone turnover, as in the case of estrone, but genistin has a different mechanism of action from the other compounds. Soy isoflavone glycosides may represent a potential alternative therapy in the treatment of bone loss and lipid metabolism abnormality in ovarian hormone-deficient women.

Garlic and onion oils inhibit proliferation and induce differentiation of HL-60 cells.

Phytochemicals present in the genus Allium have potential pharmacological effects, such as antimicrobial, antithrombotic, antitumor, hypolipidaemic and hypoglycemic activities. In this present study, we examined the effects of garlic and onion oils on human promyelocytic leukemia cells, HL-60. Incubation of HL-60 with garlic or onion oil (20 microg/ml) caused a marked suppression of HL-60 proliferation; the suppression was almost identical with those obtained by all-trans-retinoic acid (ATRA) or dimethyl sulfoxide (DMSO) used as positive controls. These oils induced the generation of nitroblue tetrazolium (NBT)-reducing activity, and about 20% of the HL-60 cells became NBT positive. CD11b, another marker of the differentiation of these cells, was also significantly induced by garlic oil or onion oil. The combination of garlic or onion oil with ATRA was more effective than either alone. These data suggest that garlic and onion oils have the ability to induce differentiation of HL-60 cells into those of the granulocytic lineage.

Studies of the active substances in herbs used for hair treatment. III. Isolation of hair-regrowth substances from Polygara senega var. latifolia TORR. et GRAY.

Four active principles, 1, 2, 3 and 4, were isolated from Polygara senega var. latifolia TORR. et GRAY by a combination of partition and column chromatography on silica gel and octadecyl silica gel (ODS), monitored by a hair-regrowth activity assay. Compounds 1, 2, 3 and 4 were identified as senegose A, senegin II, senegin III, and senegasaponin b by comparison of their spectral data with those of authentic samples.

Selective and continuous degradation of carbazole contained in petroleum oil by resting cells of Sphingomonas sp. CDH-7.

Microbial degradation of carbazole (CA), a model of hard-removal heterocyclic nitrogen compounds contained in petroleum oil, was examined using Sphingomonas sp. CDH-7 isolated from a soil sample by screening for CA-assimilating microorganisms. CDH-7 used CA as a sole source of carbon and nitrogen, and metabolized CA to ammonia via anthranilic acid as an intermediate product. When CDH-7 was cultivated in the medium containing CA at the concentration of 500 mg/l (2.99 mM), CA was completely degraded within 50 h. By the reaction with the resting cells of CDH-7, 500 mg/l of CA was completely degraded within 4 h, with 1.64 mM of ammonia accumulated in the reaction mixture. When CA was added at the concentration of 100 mg/l (0.599 mM) periodically to the reaction mixture ten times, 925 mg/l (5.54 mM) of CA was degraded within 48 h by the resting cells, and 4.50 mM of ammonia was accumulated in the reaction mixture with a 75.1% molar conversion yield based on total CA added. The resting cells could almost completely degrade CA in a two-liquid-phase system which consists of water and organic solvent, even in the presence of 20% (v/v) isooctane, n-hexane, cyclohexane, and kerosene as a model petroleum oil. In the presence of an organic solvent system such as 20% (v/v) pxylene, toluene, and heptanol, however, CA degradation yields decreased.

Studies of the active substances in herbs used for hair treatment. II. Isolation of hair regrowth substances, acetosyringone and polyporusterone A and B, from Polyporus umbellatus Fries.

Fractionation of the 50% ethanol extract of Polyporus umbellatus Fries by column chromatography on Amberlite XAD-2, silica gel, Sephadex LH-20 and octadecyl silica gel (ODS) (C18)) monitored by a hair-regrowth activity assay, afforded three active principles, 1, 2 and 3. The structures of 1, 2 and 3 were determined as acetosyringone, polyporusterone A, and polyporusterone B by comparison of their spectral data with that of authentic samples, respectively. The effects of several compounds related to acetosyringone, 3,4-dihydroxybenzaldehyde or polyporusterone A on hair regrowth were also investigated.

Neurochemical characteristics and behavioral responses to psychological stress in ovariectomized rats.

The present study was designed to clarify the time-dependent changes in brain monoamine turnover in the frontal cortex, hypothalamus, hippocampus, septum and amygdala after ovariectomy, and the difference in behavioral responses to psychological stress between sham-operated and ovariectomized (OVX) rats. At 2 and 4 weeks after ovariectomy, the turnover rates of dopamine and norepinephrine in all of the brain regions examined did not differ significantly between the sham-operated and OVX rats. However, 5-hydroxytryptamine (5-HT) turnover in all of the brain regions at 2 weeks after OVX was significantly lower than that in sham-operated rats. This difference was greater in the hypothalamus than in other brain regions. At 4 weeks after ovariectomy, 5-HT turnover in all of the brain regions examined was not significantly different between sham-operated and OVX rats. At 2 and 4 weeks after ovariectomy, exploratory behaviour (e.g., locomotor activity, head- dipping, crossing and rearing behaviours) in a non-stressed ovariectomy group did not differ from that in a non-stressed sham-operation group. Locomotor activity and the number of head-dips and crossings significantly (P<0.05) increased after repeated exposure to psychological stress for 5 days in sham-operated rats, but not in those at 2 weeks after OVX. At 4 weeks after ovariectomy, locomotor activity and the number of crossings and rearings in sham-operated and OVX rats were not significantly different in the psychological stress and non-stress groups. However, the number of head-dips significantly (P<0.05) increased with psychological stress in the sham-operated rats, but not in OVX rats. These results suggest that female gonadal hormones may play an important role in the regulation of brain 5-HTergic systems. These interactions between gonadal hormones and 5-HT metabolism may be related to 5-HT-related neuropsychiatric disorders.

Magnolol inhibits leukotriene synthesis in rat basophilic leukemia-2H3 cells.

We have observed an inhibitory action of magnolol on the production of leukotriene (LT) C4 and LTB4, important lipid mediators in allergy and inflammation. IgE- and A23187-stimulated production of LTC4 and LTB4 was measured by radio-immunoassay (RIA) in the absence or presence of various concentrations of magnolol in intact rat basophilic leukemia (RBL)-2H3 cells. Magnolol dose-dependently inhibited synthesis of LTC4 and LTB4. Magnolol inhibited the IgE-mediated increase of intracellular calcium ion concentration, resulting in the inhibition of cytosolic phospholipase A2 (cPLA2) and possibly 5-lipoxygenase (5-LO), both calcium ion-dependent enzymes. In cell-free studies magnolol inhibited LTC4 synthase activity. LTA4 hydrolase activity was only inhibited at the higher concentration (2.5 x 10(-5)M). These results indicate that magnolol inhibits production of LTs by inhibiting PLA2, 5-LO, LTC4 synthase and LTA4 hydrolase which are essential for LT-synthesis. Magnolol may have anti-allergic effect by blocking LT-synthesis.

Effects of tea polyphenols on the invasion and matrix metalloproteinases activities of human fibrosarcoma HT1080 cells.

The effects of tea polyphenols on the invasion of highly metastatic human fibrosarcoma HT1080 cells through a monolayer of human umbilical vein endothelial cells (HUVECs) and the accompanying basal membrane were investigated. Among the tea polyphenols tested, epicatechin gallate (ECg), epigallocatechin gallate (EGCg), and theaflavin strongly suppressed the invasion of HT1080 cells into the monolayer of HUVECs/gelatin membrane, whereas epicatechin, epigallocatechin, tea flavonols, tea flavones, and gallate derivatives had no effect. Both theaflavin-digallate and theasinensin D showed a weak invasion inhibitory effect. ECg significantly inhibited the invasion without cytotoxicity against cancer cells and HUVECs. Ester-type catechins (ECg and EGCg) and theaflavin strongly suppressed the gelatin degradation mediated by matrix metalloproteinase (MMP) 2 and MMP-9, which were secreted into the conditioned medium of HT1080 cells. In conclusion, among the tea polyphenols tested, ECg was considered to be the agent with the most potential antimetastasis activity because it inhibited invasion in the absence of cytotoxicity.

Hyperthermic enhancement of tumour growth inhibition by accelerated carbon-ions in transplantable human esophageal cancer.

The study examined the effects of combination of hyperthermia (42 degrees C) and 290 MeV/u carbon-ion (C-) beams or 200 kVp X-rays on tumour regrowth delay of transplantable human esophageal cancer as an in vivo model for radiotherapy of cancer. The C-beams were more effective in the tumour growth inhibition than X-rays. The relative biological effectiveness (RBE) of C-beams against X-rays was 2.00. It was observed that the interactive hyperthermic (42 degrees C, for 30 min) enhancement of tumour regrowth delay by high-linear energy transfer (LET) C-beams was similar to that of combination of low-LET X-rays with hyperthermia. The thermal enhancement ratios (TER) were 6.10 and 5.57 for X-rays and C-beams, respectively. These results suggest that hyperthermic treatment is effective in radiotherapy not only by low-LET radiation but also by high-LET radiation such as C-beams. In conclusion, the depression of the tumour growth by the combined treatment of hyperthermia (42 degrees C) and the C-beams strongly suggests the available possible application of interdisciplinary cancer therapy for refractory tumours.

Preventive effects of the plant isoflavones, daidzin and genistin, on bone loss in ovariectomized rats fed a calcium-deficient diet.

The effects of the plant isoflavones, daidzin and genistin, on bone loss in ovariectomized (ovx) rats fed a calcium-deficient diet were investigated. Daidzin and genistin were orally administered to ovx rats for 4 weeks. The femurs of these rats showed significantly lower density, strength (breaking forces), ash weight and calcium and phosphorus content (p<0.01) in comparison with those of sham-operated rats. These changes were largely prevented in animals receiving oral daidzin or genistin for 4 weeks at a dose of 50 mg/kg/d and in animals receiving subcutaneous estrone (7.5 microg/kg/d) as a positive control. Ovariectomy caused atrophy of the uterus and increased the ratio of the urinary excretion of pyridinoline and deoxypyridinoline to endogenous creatinine excretion. This was prevented by administration of daidzin or estrone, but, interestingly, not genistin. The preventive effect of daidzin treatment on bone loss in ovariectomized rats appears to be due to suppression of bone turnover. Genistin has a different mechanism of action from daidzin.

Effects of tea infusions of various varieties or different manufacturing types on inhibition of mouse mast cell activation.

We investigated effects of various tea infusions on mast cell activation using mouse mast cells. Among various tea extracts, infusions from cultivar 'Benihomare' and Taiwan lineage strongly inhibited histamine release after Fc epsilon RI cross-linking. Among three types of tea (from cultivar 'Benihomare'), extract from oolong tea or black tea inhibited histamine release more strongly than green tea extract. Furthermore, 'Benihomare' oolong tea extract suppressed tyrosine phosphorylation of cellular proteins after Fc epsilon RI cross-linking, but polyvinyl polypyrrolidone treatment of the extract to remove phenolic compounds, weakened the suppressive effect.

Isolation of inhibitors of TPA-induced mouse ear edema from Hoelen, Poria cocos.

Triterpene carboxylic acids were isolated from the methanol extract of Hoelen, Poria cocos, and found to inhibit 12-O-tetradecanoylphorbol 13-acetate (TAP)-induced mouse ear edema. Their chemical structures were identified as 3 beta,-16 alpha-dihydroxylanosta-7,9(11),24-trien-21-oic acid, 16 alpha-hydroxydehydropachymic acid, 16 alpha-hydroxytrametenolic acid and dehydrotumulosic acid.

A novel resin glycoside, merremin (tuguajalapin X dimer), from Merremia hungaiensis.

A novel resin glycoside, merremin (1), has been isolated from the root of Merremia hungaiensis (Convolvulaceae). The structure has been determined to be an ester-type dimer of tuguajalapin X (2) on the basis of chemical and spectral data.

Study on the bile salt, sodium scymnol sulfate, from Rhizoprionodon acutus. IV. Structures of chimaerol and sodium chimaerol sulfate in the bile of Lamna ditropis and Rhizoprionodon acutus.

Sodium chimaerol sulfate (1) was isolated from the bile of Lamna ditropis and Rhizoprionodon acutus by chromatography on silica gel and Sephadex LH-20, together with sodium scymnol sulfates, (24R,25S)- and (24R,25R)-(+)-3 alpha,7 alpha,12 alpha,24,26-pentahydroxy-5 beta-cholestan-27-yl sodium sulfates (3 and 4) and 3, respectively. On acid hydrolysis, compound 1 afforded chimaerol (2), which was identified as (24R,25R)-(+)-5 beta-cholestane-3 alpha,7 alpha,12 alpha,24,26-pentol by direct comparison with an authentic sample, prepared by reduction of (24R,25S)-(+)-24,26-epoxy-5 beta-cholestane-3 alpha,7 alpha,12 alpha,27-tetrol (5) with LiAlH4. The structure of 1 was concluded to be (24R,25R)-(+)-3 alpha,7 alpha,12 alpha,24-tetrahydroxy-5 beta-cholestan-26-yl sodium sulfate, based on the chemical transformation and spectral data.

Study on pharmacological effect of bile salts, sodium scymnol sulfate, from Rhizoprionodon acutus. I. Effect of scymnol, chimaerol and sodium scymnol sulfate on cerebral anoxia.

The effects of scymnol, chimaerol and sodium scymnol sulfate, prepared from the bile of Rhizoprionodon acutus, on cerebral anoxia were investigated in experimental models of hypoxia, ischemia and histotoxic anoxia in mice. Scymnol, at a dose of 100 mg/kg, showed a significant protective action against cerebral anoxia in all of the models studied and significantly increased the partial oxygen pressure of the arterial blood. The anti-anoxic activity of scymnol was found to be slightly greater than that of idevenone. A similar protective effect of sodium scymnol sulfate was seen at doses higher than 100 mg/kg. The survival time on hypoxia was significantly prolonged in the animals pretreated with chimaerol.

Resin glycosides. XXI. Tuguajalapins I-X, the resin glycosides having long-chain fatty acid groups from the root of Merremia hungaiensis.

Ten resin glycosides (jalapins) named tuguajalapins I-X were isolated from the root of Merremia hungaiensis. Their structures have been determined on the bases of chemical and spectral data. Unlike the jalapins so far reported, all of their acyl groups consist of two or three long-chain fatty acids, that is, palmitic, stearic and/or arachidic acids.