蓝莓果渣提取物总酚含量及抗氧化活性研究

研究了蓝莓果渣提取物总酚含量及其抗氧化活性。分别采用水、40%乙醇及纤维素酶辅助乙醇超声提取蓝莓果渣,并用Folin-Ciocalteu试剂对3种提取物的总酚含量进行评估;并采用DPPH清除实验及O2—.清除实验对3种提取物的抗氧化活性进行研究。实验结果表明,纤维素酶辅助超声提取蓝莓果渣的总酚含量最高,可达425.36±15.21 mg GAE.100 g-1DW,远远高于水提物(169.46±9.75 mg GAE.100 g-1DW)及醇提物(218.39±12.54mg GAE.100 g-1DW)中的总酚含量。且纤维素酶辅助乙醇超声提取物对DPPH的清除能力为2.67±0.13 gVc.100 g-1DW,对O—.2的清除能力2.48±0.14 g Vc.100 g-1DW,明显好于醇提物及水提物抗氧化活性。     

Detection of phenylalanine hydroxylase protein in human platelets. Immunochemical detection

A protein reactive with anti-phenylalanine hydroxylase monoclonal antibody PH8 has been recovered from human platelet extracts. Two bands corresponding to molecular masses of about 60 kDa and 55 kDa were revealed by immunoblotting after electrophoresis according to Laemmli. Using the same antibody, a single band with a molecular mass of 60 kDa was demonstrated in extracts from human pineal gland; two similar antigens were found in human liver extracts and no antigen was found in adrenal gland extracts. Monoclonal antibodies, PH1 and PH3, did not react with these antigens during immunoblotting. Monoclonal antibodies, PH7 and PH9, reacted with the 55 kDa antigen in platelet extracts. The antigen content in platelet extracts was measured by ELISA with monoclonal antibodies relative to its content in the liver. The antigen content in platelet extracts was about 100 times as low as that in liver extracts and amounted to 100 ng/mg of protein. These findings suggest that the phenylalanine hydroxylase antigen is present in human platelets.     

Pyruvate oxidation by the Reiter strain of Treponema phagedenis.

Spectrophotometric assays of pyruvate oxidation catalyzed by extracts of the Reiter strain of Treponema phagedenis indicated that viologen dyes, flavin nucleotides, and a ferric iron chelate, but not pyridine nucleotides, were utilized as electron acceptors. Benzyl viologen-linked activity partially sedimented during ultracentrifugation and appeared similar to clostridial pyruvate:ferredoxin oxidoreductase with respect to the spectral properties of the enzyme chromophore. Electron carrier activity in treponemal extracts was quantitated by a metronidazole-linked assay in which the oxidation of pyruvate by carrier-depleted extracts led to the reduction of electron carrier in the crude extracts which then reduced metronidazole. The rate of metronidazole reduction was proportional to the amount of electron carrier present in the assay. Electron carrier activity in Triton X-100-solubilized, crude extracts partially purified by DEAE-cellulose chromatography and gel filtration was attributed to a protein possessing the spectral and physical properties of a ferredoxin. A similar protein appeared to be present in extracts of Treponema denticola ST10.     

Antimutagenic effect of eight natural foods on moldy foods in a high liver cancer incidence area.

Ames test procedures were used to test 8 natural food extracts for their antimutagenic activity against the mutagenic activity induced in S. typhimurium strains TA98 and TA100 by aflatoxin B1 (AFB1) or metabolic extracts from A. versicolor or A. ochraceus. The tested substances were extracted repeatedly with acetone. The revertants induced by AFB1, metabolic extracts of A. versicolor or A. ochraceus were significantly decreased when extracts of the 8 natural foods were added to the media. The results showed that these extracts had marked inhibitory effects on the mutagenic activity induced by AFB1 or metabolic extracts of the two molds and also suggested that antimutagenic substances were present in these natural foods. These experiments provide a scientific basis for the study of food substances for the prevention of carcinogenesis. It is considered that these 8 natural food extracts produce marked antimutagenic effects and are practically valuable in the field of chemoprophylaxis of liver cancer in humans.     

Evaluation of antidepressant-like activity of aqueous and ethanolic extracts of Terminalia bellirica Roxb. fruits in mice

The present study was undertaken to investigate the effect of aqueous and ethanolic extracts of T. bellirica on depression in mice using forced swim test (FST) and tail suspension test (TST). The extracts were administered orally for 10 successive days in separate groups of Swiss young male albino mice. Aqueous extract (50, 100 and 200 mg/kg) in a dose-dependent manner and ethanolic extract (100 mg/kg) significantly reduced the immobility time of mice in both FST and TST. The extracts were without any significant effect on locomotor activity of mice. The efficacies of aqueous extract (200 mg/kg) and ethanolic extract (100 mg/kg) were found to be similar to that of imipramine (15 mg/kg, po) and fluoxetine (20 mg/kg, po) administered for 10 successive days. Both extracts reversed reserpine-induced extension of immobility period of mice in FST and TST. Prazosin (62.5 microg/kg, ip; an alpha1-adrenoceptor antagonist), sulpiride (50 mg/kg, ip; a selective D2 receptor antagonist) and p-chlorophenylalanine (100 mg/kg, ip; an inhibitor of serotonin synthesis) significantly attenuated the aqueous and ethanolic extract-induced antidepressant-like effect in TST. Thus, both the aqueous and ethanolic extracts of T. bellirica elicited a significant antidepressant-like effect in mice by interaction with adrenergic, dopaminergic and serotonergic systems.     

Mutagenicity of processed bidi tobacco: possible relevance to bidi industry workers

The genotoxic potential of bidi tobacco was evaluated by mutagenicity testing of aqueous, aqueous: ethanolic, ethanolic and chloroform extracts of processed tobacco used in the manufacture of 'bidis', indigenous forms of cigarettes smoked in India. The Salmonella/mammalian microsome test (Ames assay) was used to detect mutagenicity in tester strains TA98, TA100 and TA102. The extracts were tested in the absence and presence of metabolic activation using liver S9 from rat and hamster, and following in vitro nitrosation with sodium nitrite at acidic pH. All the extracts were non-mutagenic in the absence of nitrosation. The nitrosated aqueous extract was mutagenic in strains TA98 and TA100. While weak mutagenicity was elicited by the nitrosated aqueous: ethanolic extract in TA100, the nitrosated ethanolic extract induced a 3-fold increase in the number of revertants in the same strain. Moreover both these extracts elicited a strong mutagenic response in TA102, while the chloroform extract was non-mutagenic even after nitrite treatment. The present study indicates that workers employed in the bidi industry are exposed to potentially mutagenic and genotoxic chemicals in the course of their occupation.     

Potentiation of pentobarbital hypnosis by Rosa damascena in mice

Rosa damascena has been found to act on central nervous system including brain. It inhibits the reactivity of the hypothalamous and pituitary systems in rat. In traditional medicine hypnotic effect of Rose is also suggested. In the present study hypnotic effect of ethanolic, aqueous and chloroformic extracts of R. damascena was investigated in mice. Hypnotic method was based on potentiation of pentobarbital induced sleeping time by extracts. Three doses of extracts (100, 500 and 1000 mg/kg) were injected i.p. in comparison with diazepam (3mg/kg) as positive control and saline as negative control. After 30 min of injection of extracts, pentobarbital (30mg/kg) was injected and increase in sleeping time by extracts was recorded. The results showed that the ethanolic and aqueous extracts in 500 and 1000 mg/kg doses significantly increased pentobarbital induced sleeping time which was comparable to diazepam. The chloroformic extract had no hypnotic effect.     

Effects of Lavandula officinalis and Equisetum arvense dry extracts and isoquercitrin on the fermentation of diets varying in forage contents by rumen microorganisms in batch culture

The short-term actions of Lavandula officinalis and Equisetum arvense dry extracts, and of isoquercitrin, flavonoid present in Equisetum arvense, on in vitro fermentation by rumen microbes were studied in batch culture. The orchard grass hay:barley ratios in the three experimental diets were 100:0, 75:25, 50:50 on a DM basis. The production rates of all volatile fatty acids except isobutyrate were strongly influenced by the composition of the diet and to a lesser extent, by plant extracts, with significant interactions between both factors. When hay was the only substrate, the addition of L. officinalis and E. arvense enhanced the fermentation rate by 50%, through an increased release of acetate and propionate. On the contrary, with the two other diets, the fermentation rate was strongly lowered by isoquercitrin. Gas outputs were not significantly influenced by plant extracts.     

Comparative assessment of solvents and lignocellulolytic enzymes affiliated extraction of polyphenols from the various lignocellulosic agro-residues: identification and their antioxidant properties

Abstract

The present investigation was aimed to utilize lignocellulosic agro-residues and compare the extraction of polyphenols utilizing lignocellulolytic enzymes secreted by Sphingobacterium sp. ksn and with that of the solvents (ethanol, methanol) affiliated methods. The maximum amount of polyphenols, flavonoids and tannins were 94.29, 11.36, and 79.21?g 100?g^?1 respectively, found in the extracts obtained by enzymes affiliated extraction of coffee cherry husk (CCH). The phenolics namely, gallic acid, caffeic acid, coumaric acid, 1-hydroxybenzoic acid, 2,5-dihydroxybenzoic acid, p-hydroxybenzaldehyde were commonly found whereas syringic acid, quercetin, kaempferol, and epicatechin were hardly found in the extracts of agro-residues. The extracts of CCH shown maximum antioxidant properties for DPPH, ABTS, and FRAP. The present study reports that the affiliation of enzymes for the extraction of polyphenols from agro-residues is more efficient than that of the solvents affiliation and CCH as the good source of polyphenols.     

Oral mucosal pellicle. Adsorption and transpeptidation of salivary components to buccal epithelial cells.

The present investigation was carried out to examine the mechanism(s) whereby salivary molecules interact with human buccal epithelial cells. By utilizing antiserum against human parotid saliva, selected salivary components were detected by electrophoretic-transfer analysis of 1.5% SDS extracts of epithelial cells. Incubation of the cells and their aqueous cell-free extracts with 125I-labelled parotid saliva resulted in the formation of an iodinated high-molecular-mass complex which was not present in 125I-labelled saline alone. Formation of this complex was time-dependent and was inhibited by treating the buccal epithelial cells or their cell-free extracts with EGTA, iodoacetamide, N-ethylmaleimide or by heating at 100 degrees C for 15 min. The epithelial cells also promoted incorporation of [14C]putrescine into high-molecular-mass complexes whose formation was inhibited by iodoacetamide, unlabelled putrescine and EGTA. Cell extracts mediated cross-linking of monodansylcadaverine into alpha-casein, and this interaction was inhibited by iodoacetamide. Significant amounts of radioactivity were recovered with the epithelial-cell envelopes after exhaustive extraction of 125I-saliva- or [14C]putrescine-treated epithelial cells with 4% (w/v) SDS/10% (v/v) beta-mercaptoethanol. The incorporation of radioactivity into epithelial-cell envelopes was inhibited by pretreatment of the cells with putrescine, EGTA, iodoacetamide, or heating at 100 degrees C for 15 min. These data suggest that: (1) oral mucosal pellicle is formed by the selective adsorption of saliva to the epithelial-cell plasma membrane and its associated cytoskeleton; and (2) the adsorbed salivary components may be cross-linked to each other or the epithelial cytoskeleton by epithelial transglutaminases.     

Comparison of cell surface antigen extracts from two serotypes of Pasteurella haemolytica

Cells of Pasteurella haemolytica serotypes A1 and A6 were extracted with sodium salicylate and the chemical and antigenic composition of both extracts determined. The extracts were concentrated by ultrafiltration and the serotype antigen, measured by the indirect haemagglutination test, was estimated to have a molecular weight between 100 000 and 300 000. The chemical composition of sodium salicylate extracts (SSEs) from both serotypes was similar, having protein, carbohydrate, fatty acid and phosphorus present in the ratio 10:1:0.5:0.1. SDS-PAGE of both SSEs gave similar profiles with at least 48 bands present. These results suggest that sodium salicylate removes the outer membrane of P. haemolytica. Crossed immunoelectrophoresis indicated that a major serotype-specific antigen was present in SSEs of both strains. This antigen was extracted from the SSE with hot phenol/water and analysed by gas chromatography. The sugar composition of A1 and A6 phenol/water extract (PWE) was qualitatively identical although some differences in proportions were observed. A1 and A6 PWE antigens protected mice against homologous serotype challenge and A6 PWE protected against heterologous (A1) challenge.     

Detection of oxidative mutagens in an urban air-particulate extract: a preliminary study.

The Ames assays strains TA98 and TA100 have been useful in characterizing complex mixtures from organic solvent extracts of particles from diesel-powered vehicles, ambient air, and other sources. In this paper we report preliminary experiments using TA102, a bacterial strain that detects compounds that can oxidize DNA, to characterize the mutagenicity of an ambient air sample collected in Ann Arbor, MI. Four sets of ambient air filters were collected in duplicate over a period of several days. The mutagenicities of methylene chloride extracts of these filters were compared using strains TA98, TA100 and TA102. The concentration-mutagenicity data for TA98 and TA100 were linear over the concentration range 0-200 micrograms extract/plate. The mutagenicity of the extracts using TA102 was much lower than the other two strains and was non-linear over the concentration range tested. These results suggest that it would be difficult to use TA102 to identify the oxidative mutagens present in an ambient air particulate extract.     

Effect of seed extracts of <Emphasis Type="Italic">Withania somnifera,Croton tiglium</Emphasis> and <Emphasis Type="Italic">Hygrophila auriculata</Emphasis> on behavior and physiology of <Emphasis Type="Italic">Odontotermes obesus</Emphasis> (Isoptera,Termitidae)

In addition to antibiotic properties, medicinal plants are important sources of chemicals with potential application as pesticides. The present study deals with antitermitic potential of seed extracts of Withania somnifera (Indian ginseng), Croton tiglium (jamalgoota) and Hygrophila auriculata (talimkhana). The seed extracts caused changes in tunneling behaviour, number of bacterial colonies in hindgut and activities of enzymes in midgut of Odontotermes obesus. C. tiglium showed the lowest LT₅₀ (12.85 and 2.65 h) among the three seed extracts at concentrations of 50% (half dilution of the extract) and 100% (extract without dilution), respectively. There was no tunneling in soil treated with 100% concentration of seed extracts of W. somnifera and C. tiglium. Numbers of bacterial colonies in the gut of termites from soils treated with 50% and 100% concentrations of the three plants did not differ significantly, but they differed from those in termites from untreated soil. At 50% concentrations of seed extracts of the tested plants, the difference in hindgut enzyme activities was not obvious, however, at 100% concentrations the enzyme activities in the termites from soils treated with seed extracts significantly differed from controls and differences were also recorded between the plants.     

Inhibitor of diamine oxidase in cotyledons of groundnut seedlings

The apparent absence of diamine oxidase in extracts of cotyledon of germinating groundnut seeds is due to the presence of a natural inhibitor. The inhibitor was associated with the lipid in the upper layer obtained on centrifugation of the cotyledon extract. It was non-dialysable, thermolabile and was inactivated by TCA and Triton X-100. The inhibitor was detected in the cotyledon extracts from early stages of seed development and was present up to 20 days after germination.     

Characterization of homologous DNA recombination activity in normal and immortal mammalian cells.

Homologous DNA recombination levels were measured in normal and spontaneously immortalized murine and human fibroblasts, and in a number of primate and murine established fibroblast cell lines. Immortal cell lines and tumor-derived clones homologously recombined extrachromosomal plasmid substrates at frequencies approximately 100-fold higher than did normal cells. To further explore the mechanism responsible for this phenotype, homologous recombination frequency was measured using nuclear extracts derived from normal and immortalized murine and human fibroblasts. Extracts prepared from immortal cells catalyzed high levels of homologous recombination, whereas very little recombination activity was detected in extracts prepared from normal fibroblasts. Similarly, only extracts derived from immortal cells contained strand-transferase activity as measured by the recently described pairing-on-membrane assay. Mixing experiments indicated that a recombination enhancing factor or factors present in immortal cells, rather than a recombination inhibitor in normal cells, was responsible for the enhanced homologous recombination activity observed using extracts derived from the former.     

Ginseng extract exhibits antimutagenic activity against induced mutagenesis in various strains of Salmonella typhimurium

Ginseng has been reported to exhibit antioxidant and antimutagenic activity. The present study was undertaken with a view to confirm whether the antioxidant activity of Ginseng is responsible for its antimutagenic action. The concentrated root extract of Panax ginseng (Ginseng extract I) and its lyophilized powder (Ginseng extract II) obtained from two different manufacturing houses, were tested against mutagenesis using the well-standardized Ames microsomal test system. The extracts exhibited antimutagenic effect against hydrogen peroxide induced mutagenesis in TA100 strain, and against mutagenesis produced by 4-nitroquinoline-N-oxide in both TA98 and TA100 strains of Salmonella typhimurium. Both the extracts failed to show any antimutagenic potential against tert-butyl hydroperoxide (an oxidative mutagen) in TA102 strain, a strain highly sensitive to active oxygen species. The extracts also indicated a weak antioxidant activity in a series of in vitro test systems viz., 1,1-diphenyl picryl hydrazyl (DPPH) assay, hydrogen peroxide scavenging and superoxide anion scavenging. The results indicate that the protective effects shown by ginseng extract(s) against 4-nitroquinoline-n-oxide and hydrogen peroxide induced mutagenesis in TA98 and TA100 could mainly be due to its property to initiate and promote DNA repair rather than free radical scavenging action.     

Immunoelectrophoretic analysis of erythrocytic stages of Plasmodium yoelii and P. chabaudi

Triton X-100 extracts of erythrocytes infected with Plasmodium chabaudi and P. yoelii were analysed in crossed immunoelectrophoresis with rabbit antisera. The parasite origin of the antigens detected was assessed by metabolic radiolabelling of the parasites with 35S-methionine. About 12 immunoprecipitates were obtained with both extracts and their homologous antiserum. Cross-tests showed that the two parasite strains were very similar antigenically. Species-specific antigens could, however, also be demonstrated. Two antigens, present on both P. yoelii- and P. chabaudi-infected erythrocytes, were located on the surface of the host cell membrane as judged from 125I-labellings with lactoperoxidase. Experiments with phenyl-Sepharose showed that most of the antigens detected were hydrophilic and none of them reacted with concanavalin A.     

杠柳根皮乙醇提取液对蔬菜害虫小菜蛾的生物活性

采用95%乙醇对杠柳(Periploca sepium Bunge)根皮进行热提取,以叶片浸渍法和点滴法测定了提取液对小菜蛾(Plutella xylostella L.)的杀虫活性及其作用方式.结果显示,杠柳乙醇提取液稀释100倍处理对小菜蛾3龄和4龄幼虫24 h后的非选择性拒食率分别为87.3%和96.3%;100倍液浸叶饲喂处理对小菜蛾2龄幼虫72 h后的校正死亡率为80%,对小菜蛾3龄幼虫24 h和48 h后的生长抑制率为100%.杠柳乙醇提取液对小菜蛾幼虫具有较高的生物活性,其作用方式包括拒食作用、胃毒作用和生长抑制作用.此外,乙醇提取液对小菜蛾幼虫还有一定的触杀和内吸效应,并对小菜蛾成虫产卵有明显的忌避活性,但对小菜蛾卵没有杀伤作用.     

Cyclitol utilization associated with the presence of Klebsielleae in botanical environments.

Bacteria of the tribe Klebsielleae are capable of metabolizing the cyclitols myoinositol, sequoyitol, and pinitol, which are present in aqueous extracts of redwood. Of the combined Klebsiella isolates from clinical and environmental origins, 100% (138/138), 97% (34/35), and 86% (119/138) fermented inositol, sequoyitol, and pinitol, respectively. These compounds were also used as a sole source of carbon and energy by Klebsiella. Similar results were obtained with Enterobacter isolates, but most other enteric bacteria tested could not metabolize cyclitols. Strains of Klebsiella multiplied to levels exceeding 10(5)/ml in aqueous extracts of nonsterile redwood within 6 days. Most other enteric bacteria did not grow in these extracts. Cyclitol metabolism was found to correlate well with the ability to multiply in redwood extract in the presence of cyclitol-negative indigenous bacteria. The capacity to use cyclitols, which are present in a variety of plant material, might afford Klebsielleae of both environmental and clinical origins an advantage in competing for nutrients and colonizing botanical environments.     

Cyclitol utilization associated with the presence of Klebsielleae in botanical environments.

Bacteria of the tribe Klebsielleae are capable of metabolizing the cyclitols myoinositol, sequoyitol, and pinitol, which are present in aqueous extracts of redwood. Of the combined Klebsiella isolates from clinical and environmental origins, 100% (138/138), 97% (34/35), and 86% (119/138) fermented inositol, sequoyitol, and pinitol, respectively. These compounds were also used as a sole source of carbon and energy by Klebsiella. Similar results were obtained with Enterobacter isolates, but most other enteric bacteria tested could not metabolize cyclitols. Strains of Klebsiella multiplied to levels exceeding 10(5)/ml in aqueous extracts of nonsterile redwood within 6 days. Most other enteric bacteria did not grow in these extracts. Cyclitol metabolism was found to correlate well with the ability to multiply in redwood extract in the presence of cyclitol-negative indigenous bacteria. The capacity to use cyclitols, which are present in a variety of plant material, might afford Klebsielleae of both environmental and clinical origins an advantage in competing for nutrients and colonizing botanical environments.