Novel DPPH Radical Scavengers,Demethylbisorbibutenolide and Trichopyrone,from a Fungus

In our screening program for antioxidants with 1,1-diphenyl-2-picrylhydrazyl (DPPH)-radical scavenging activity, two novel compounds, demethylbisorbibutenolide (1) and trichopyrone (2), were isolated from the fermentation broth of the fungus of USF-4860 strain isolated from a soil sample. The structures of these compounds were determined from spectroscopic evidence. The biosynthetic origin of the carbon atoms of 2 was unambiguously determined by feeding experiments using ¹³C-labeled precursors and elucidation of the ¹³C-NMR spectrum of ¹³C-labeled 2. These studies showed that 2 was derived from five acetates and a methyl group of methionine. In the DPPH-radical scavenging assay, 1 and 2 gave ED₅₀ values of 149 and 167 μM after standing for 2.0 hr. Compound 2 reacted with the DPPH radical to form reaction product 3 which was determined to be 1-[4-(3,4-dihydro-3-methyl-6-{1,3-pentadienyl}-2,4-dioxo-2H-pyran-3-yl)-phenyl]-1-phenyl-2-picrylhydrazine from spectroscopic evidence.     

Ansaetherone,a New Radical Scavenger from Streptomyces sp.

We identified a new radical scavenger, ansaetherone (C₂₆H₃₃NO₇), from a culture of the Streptomyces sp. USF-4727 strain. In our previous study, it was shown that this strain produced four lipoxygenase inhibitors, tetrapetalones A, B, C and D. The chemical structure of ansaetherone was elucidated by the spectroscopic method, indicating that this compound was constructed with an aglycon and a sugar moiety. This chemical structure suggested that ansaetherone was related to the tetrapetalones. This finding provided information regarding tetrapetalone biosynthesis. Ansaetherone showed radical scavenging activity with an ED₅₀ value of 300 μM in our assay.     

Identification of the reduced primary electron acceptor of Photosystem II as a bound semiquinone anion

The complete absorption difference spectrum of the primary electron acceptor of Photosystem II has been measured at room temperature in subchloroplast fragments prepared with deoxycholate. The shape and amplitude of the spectrum indicate that the primary reaction involves the reduction of one bound plastoquinone molecule per reaction center to its semiquinone anion. In addition two small absorbance band shifts occur near 545 (C550) and 685 nm, which may be due to an influence of the semiquinone on the absorption spectrum of a reaction center pigment.     

Synthesis and Insecticidal Activity of Isobutenyl- and Isobutyl-cyclopropanecarboxylates

Several isobutenyl- and isobutyl-cyclopropanecarboxylates were synthesized. The insecticidal activities against the housefly of their 5-benzyl-3-furylmethyl or allethronyl esters were tested. Among the substituents on the cyclopropane ring, a methyl group cis to the ester linkage has been found to be essential for toxicity against the housefly and a trans-isobutenyl group greatly enhances the toxic activity.     

2-Ketogluconate Reductase Inhibition by Oxalate

The active site of α-glucosidase from Mucor javanicus IFO 4570 was investigated by kinetic studies. Competition between maltose and soluble starch, and linearity of Lineweaver-Burk plots for the mixed substrates were observed. The dependence of the apparent maximum velocities agreed with those predicted for a single active site mechanism. These results suggest that the enzyme hydrolyzes maltose and soluble starch at a single active site.     

The Occurrence and Some Properties of a Spawning Inducing Substance in the Testis of the Hydromedusan Spirocodon saltatrix

Gas chromatographic methods were developed for the determination of optical isomers of 2-(4-chlorophenyl)isovaleric acid (CPIA) which constituted the acid moiety of the insecticide Fenvalerate molecule. The enantiomers of CPIA were derivatized into diastereoisomeric l-menthyl esters quantitatively via their acid chlorides and separated from each other on a column of 10% silicone DC QF-1 (3 mm i.d. × 2.25m). They were also derivatized into isopropylamides in the presence of N,N′-dicyclohexylcarbodiimide and were resolved on an open tubular glass capillary column (0.25 mm i.d. × 40 m) coated with optically active N,N′-[2,4-(6-ethoxy-1,3,5-triazine)diyl] bis(l-valyl-l-valyl-l-valine isopropyl ester) (OA-300). The ratios of optical isomers were determined from their separated peak areas and analytical values obtained by two methods were in good agreement with each other.

The chemical purity of CPIA was also determined by gas chromatography after derivatization to its methyl ester on a column of 2% DEGS (3 mm i.d. × 3 m) using m-nitroanisole as an internal standard.     

三种黑米花色苷自由基清除能力的理论评价

用一种组合的密度泛函理论方法计算了三种黑米花色苷的O-H键解离焓和电离势,以此作为理论指标评价其自由基清除能力。计算结果发现花色苷的电离势和总抗氧化能力存在良好的线性关系,提示电离势表征的质子转移伴随电子转移的反应机制在清除自由基的过程中发挥着重要作用。     

Physiological Activities of Zeanic Acid,a New Plant-growth Promotor from Corn Steep Liquor

Increase in fruit set of grape was observed when vines were sprayed with zeanic acid before blossom, but when sprayed after blossom the effect was not clear.

Promotion of growth of radish cotyledons by zeanic acid was observed in the presence and absence of kinetin. Zeanic acid also promoted growth of dwarf mutant of rice seedlings (cv. Tanginbozu), but inhibited that of normal seedlings (cv. Tonewase). Zeanic acid showed promotive effect on proliferation of rice callus tissue in the absence of kinetin and auxin.

Zeanic acid showed low toxic activity against mice, earthworm and killifish, and no antimicrobial activity.     

Effects of membrane-stabilizing agents,cholesterol and cepharanthin,on radiation-induced lipid peroxidation and permeability in liposomes

Effects of two membrane-stabilizing agents, cholesterol and cepharanthin, on radiation-induced lipid peroxidation and membrane permeability were examined. Radiation-induced lipid peroxidation caused an increase in membrane permeability in phosphatidylcholine liposomes. The presence of cholesterol in liposomal membranes caused a decrease in the degree of membrane permeability in spite of an increased lipid peroxidation. On the other hand, cepharanthin suppressed both lipid peroxidation and the changes in permeability induced by radiation. The membrane-stabilizing effect of cholesterol against radiation-induced changes in permeability seemed to depend on the rigidification of membranes, which was estimated by ESR studies. Cepharanthin suppressed the degree of membrane permeability mainly by inhibiting the radiation-induced lipid peroxidation. However, cepharanthin did not exhibit a radical-trapping ability.     

7种广西产甜茶抗氧化活性的比较

通过清除二苯代苦味酰基(1,1-diphenyl-2-picrylhydrazyl,DPPH)法、铁离子还原测定(ferric reducing antioxidant power,FRAP)法、亚铁离子螯合(ferrous ion-chelating,FIC)法、β-胡萝卜素-亚油酸法等4种体外抗氧化活性测定方法,首次比较了3种茶科属茶叶和7种广西产甜茶水提物的抗氧化活性,同时用Folin-Ciocalteus法测定了其总多酚的含量。研究结果表明,多穗柯甜茶和藤茶的总多酚含量高于其他供试品。藤茶具有很好的清除DPPH·的能力,高于绿茶及阳性对照水溶性维生素E(Trolox)和人工合成抗氧化剂二丁基羟基甲苯(butylated hydroxytoluene,BHT)。甜叶菊茶和悬钩甜茶的金属离子螯合能力均高于绿茶。其中,黄杞和牛白藤两种植物的抗氧化活性为首次报道。由此可见,广西产甜茶均具有较高的开发利用价值,是治疗和防治慢性代谢性疾病的重要研究对象。对广西产甜茶的研究对于了解和抢救某些少数民族的历史文化具有重要意义,为别样茶的抗氧化研究奠定了基础,值得进一步探索和开发。     

色钉菇子实体化学成分及其生物活性初探

从色钉菇Chroogomphus rutilus子实体中分离纯化出11种化合物,经核磁等方法鉴定为：（1）麦角甾-7,22-二烯-3-酮;（2）麦角甾-4,6,8(14), 22-四烯-3-酮;（3）邻苯二甲酸二丁酯;（4）顺式-3-己烯醇;（5）3β,5α,6β-三羟基麦角甾-7,22-二烯;（6）α-甜没药醇;（7）2-甲氧基腺嘌呤核苷;（8）(4E,8E)-2-N-(2-羟基棕榈酰)-1-O-B-D-吡喃葡萄糖基-9-甲基-4, 8-sphingadienine;（9）5-羟基尿嘧啶核苷;（10）2-氨基-3-醛基-6-甲氧基吡啶;（11）Polyozellin。对分离化合物的体外抗氧化、抗肿瘤活性的初步测定结果显示,2-甲氧基腺嘌呤核苷、顺式-3-己烯醇表现出较强的清除DPPH自由基活性,EC50值分别为0.06128mg/mL和0.08253mg/mL。5-羟基尿嘧啶核苷对人胃癌细胞株BGC的增殖有较强的抑制作用,IC50值为7.92μg/mL。研究结果为进一步开发利用色钉菇提供了科学依据。     

Antimicrobial, antioxidant, and antiviral activities of Retama raetam (Forssk.) Webb flowers growing in Tunisia

Antimicrobial, antioxidant, and antiviral activities of flower extracts of Retama raetam Forssk. Webb (Fabaceae) were screened both from standard and isolated Gram-positive and Gram-negative bacteria by solid medium technique. Oxacillin, Amoxicillin, Ticarcillin, Cefotaxim, and Amphotericin were used as the control agents. The antiviral activity was evaluated against human cytomegalovirus (HCMV) strain AD-169 (ATCC Ref. VR 538) and coxsackie B virus type 3 (CoxB-3) using a cytopathic effect (CPE) reduction assay. The antioxidant activity was evaluated using two tests: 1,1-diphenyl-2-picrylhydrazyl (DPPH) free-radical scavenging and the ammonium thiocyanate methods. All extracts were characterized quantitatively for the presence of polyphenols, flavonoids, and tannins. Of the extracts tested, butanol and ethyl acetate extracts showed important antibacterial activity against Gram-positive and Gram-negative bacteria but only moderate antifungal activity. Methanol extract exhibited moderate antiviral activity against HCMV with IC₅₀ of 250 μg/ml. Ethyl acetate, chloroform, and methanol fractions were found to cause significant free-radical-scavenging effects in both assays. These results may suggest that R. raetam flowers could be used as a natural preservative ingredient in the food and/or pharmaceutical industries.     

Synthetic Analogues of the Termite Trail-following Substance

The effect of tripropyltin chloride (TPT) on some functional reactions in E. coli was investigated. The inhibition on respiration and protein, DNA and RNA syntheses were examined in vivo. Oxygen consumption by E. coli cells was scarcely inhibited even at the concentration of 30 µg/ml TPT. The incorporations of ¹⁴C-labeled amino acids into protein fraction were inhibited. Especially, in the case of l-leucine, it was inhibited 60% even at 10 µg/ml TPT. Both incorporations of ¹⁴C-adenine into DNA and RNA fraction were inhibited 50–60% at 20 µg/ml TPT. RNA polymerase was prepared from E. coli cells and the effect of organotin compounds on the enzyme activity was examined. Organotin compounds inhibited the enzyme activity only at high concentrations (5-10mm). and dialkyltin chlorides which possess no antimicrobial action showed the inhibition more intensely than trialkyltin chlorides. The effect on membrane-bound ATPase was also examined in vitro. We found that TPT has high inhibitory action on membrane-bound ATPase. However, it slightly inhibited the activity of ATPase separated from membrane.     

Preventive Effect of Lactobacillus delbrueckii subsp. bulgaricus on the Oxidation of LDL

Lactobacillus delbrueckii subsp. bulgaricus 2038 was examined for its activity to prevent the oxidation of the erythrocyte membrane in vitro, and the oxidation of LDL in vivo.

Strain 2038 produced radical scavengers that reacted with 1,1-diphenyl-2-picrylhydrazl (DPPH) during cultivation. Moreover, the ethereal extract from the supernatant of the culture prevented the oxidation of the erythrocyte membrane in vitro.

As an in vivo study, male F344 rats were fed on diets containing 20% fresh soybean oil (or 13% oxidized oil and 7% fresh oil) with 10% freeze-dried powder of the 2038 culture (or with skim milk powder) for 4 weeks. The level of thiobarbituric acid-reactive substances was lower in the low-density lipoproteins (per milligram of cholesterol) from rats fed on the oxidized oil with freeze-dried powder of the 2038 culture than without it. The level of vitamin E in the plasma was higher in the rats fed on the oxidized oil with the freeze-dried powder than without it.     

一些裸子植物鲜叶提取物清除自由基的活性研究

用二苯基苦基苯肼自由基酶标仪法,对常见的60余种裸子植物鲜叶的自由基清除活性进行了比较.发现不同科属,不同树种鲜叶的80%甲醇提取物的自由基清除活性有很大差异,其中罗汉松科罗汉松属树木鲜叶在浓度在0.5mg/mL,于37℃下孵育20min时的自由基清除率平均可达55.9%,而柏科侧柏属平均仅为7.1%;罗汉松,孔雀柏、小叶罗汉松、湿地松、杉木和刺柏等鲜叶有很强的自由基清除活性,它们的自由基清除率分别可达61.1%,58.4,57.6%,57.1%,51.8%和50.4%,表明这些树种有较大的开发潜力。     

Synthesis and Cytotoxic Activity of Thiazolofluorenone Derivatives

The synthesis and biological evaluation of some novel thiazolofluorenones, thiazolofluorenes and thiazoloanthraquinones, substituted with amino side-chains are described. These polyheterocyclic compounds have been synthesized via the corresponding imino-1,2,3-dithiazoles. Their cytotoxic activity and their eventual selective effect on a phase of the cell cycle were evaluated in vitro, using the murine lymphocytic L1210 leukaemia cell line.     

Biodegradation of the chlorophenoxy herbicide (R)-(+)-mecoprop by Alcaligenes denitrificans

An Alcaligenes denitrificans strain capable of utilizing theherbicide (R)-(+)-2(2-methyl-4-chlorophenoxy)propionicacid (mecoprop) as a sole carbon source was isolated fromsoil and cultured in liquid medium. Crude cell extracts of thebacterium were utilized in spectrophotometric assays toelucidate a biochemical pathway for degradation ofmecoprop. Results indicated a reaction sequence analogousto the degradation of 2,4-dichlorophenoxyacetic acid (2,4-D).GC-MS analysis provided direct evidence for thebiotransformation of mecoprop to the transient metabolite4-chloro-2-methylphenol (MCP). No NADPH-dependentactivity was observed during this reaction. Pyruvate wasverified as the second product derived from the aliphatic sidechain of mecoprop. MCP was subsequently transformed to asubstituted catechol by an NADPH-dependentmonooxygenase. When grown on mecoprop, A.denitrificans was adapted to oxidize catechol and its 4- and3-methylated derivatives indicating the broad substratespecificity of catechol dioxygenase. The microorganism wasdemonstrated to adopt the ortho mechanism of aromaticcleavage which resulted in the formation of2-methyl-4-carboxymethylene but-2-en-4-olide, a reactionintermediate of the -ketoadipate pathway.     

一些单子叶木本植物鲜叶提取物清除DPPH自由基活性初探

研究用二苯基苦基苯肼自由基酶标仪法,对常见的三十余种单子叶木本植物鲜叶的自由基清除活性进行了比较,发现不同科属、不同种植物鲜叶的80％甲醇提取物对DPPH自由基的清除活性有很大差异,其中供试的棕榈科7种植物鲜叶提取物,在相当于鲜叶浓度为2．5mg／ml于37℃下孵育20min时,对0．5mmol／L DPPH自由基清除率平均可达40．0％,而龙舌兰科5种植物平均仅为7．2％;刺葵、棕竹、筋头竹、蒲葵和棕榈等鲜叶有较强的自由基清除活性,它们在相当于鲜叶浓度为2．5mg/ml时的自由基清除率分别可达83．1％、79．2％、64．9％、60．5％和51．3％。这些树种有一定的开发潜力。     

亚热带一些常见木本油料树种鲜叶提取物清除自由基的活性初探

用二苯基苦基苯肼自由基酶标仪法,对亚热带常见的90余种油料树种鲜叶80％甲醇提取物的自由基清除活性进行了比较,发现所有树种提取物的自由基清除率都随浓度的增加和随着37℃下孵育时间的延长而增大。其中,苦茶槭、金缕梅、黄连木、茶、三角枫、黄山栾树、山麻杆、红瑞木、槛木、秀丽槭、算盘子、木蜡树、油茶、米心水青冈、乌柏、栾树和山茶等树叶提取物在相当于鲜叶浓度O．5mg／mL时,37℃下孵育20min后的自由基清除率分别达96．7％、93．2％、9o．6％、87．5％、86．4％、85．O％、84．3％、82．1％、80．8％、77．1％、74．5％、72．5％、72．3％、68．7％、63．8％、62．O％和61．7％。这些树种有较大的开发潜力。