Antifeeding Active Substances for Insect in Plant

The survey of the presence of chemical resistant factors in plants against the larvae of Spodoptera litura F. was examined. In addition, the antifeeding diterpenes were surveied from thirteen species of plants that belong to Verbenaceae family. Finally isolated thirteen antifeedants and one derivative were examined the antifeeding activity for the larvae by the leaf disk method.     

Effect of Humic Substances on the Growth of Microalgal Cultures

Treatment with humic substances (preparation Biomin) considerably increased viability of algae during long storage (6 months) in collection. The effect was greater with green algae (Scenedesmus and Chlorella) than with blue-green algae (Nostoc and Anabaena). In intensive cultures, lower Biomin concentrations (1–10 mg/l) increased the biomass production, protein, chlorophyll and carotenoid content, as well as -esterase and glutamate dehydrogenase activities, whereas the lipid and carbohydrate content was higher at 100 mg/l Biomin. A high Biomin concentration (1 g/l) exerted an inhibition. Biomin supply to the cultures in outdoor installation of Shetlik type was associated with a moderate stimulation of algal biomass accumulation. It was concluded that Biomin might be helpful in the improvement of algal viability and growth.__________From Fiziologiya Rastenii, Vol. 52, No. 3, 2005, pp. 463–466.Original English Text Copyright © 2005 by Pouneva.This article was submitted by the author in English.     

谷子胚性愈伤组织粘液提取物对原生质体培养的影响

将谷子胚性愈伤组织粘液提取物添加到谷子原生质体培养基中,其对原生质体培养的影响表明该提取物有助于原生质体形成细胞壁;并且该类有粘液分泌的念伤组织的原生质体游离所需的酶液浓度低、处理时间短。由原生质体形成完整细胞的数量在一定范围内与谷子原生质体培养的植板率相对应;通过增加形成完整细胞的数量可较大幅度地提高原生质体培养的植板率。     

Studies on Insect Growth Regulating Substances with Insect Cell Cultures

The sensitivity of cultured insect cells to a moulting hormone depended on their origin. Cell proliferation of cell-line C6/36, which originated from an Aedes albopictus larva, was suppressed by high concentrations of 20-hydroxyecdysone but was greatly promoted by low concentrations. Similar phenomena were observed with extracts of cedar and pine pollen. By employing the C6/36 cell-line for the screening of insect growth regulating substances, a highly active cell-growth promoting substance was found in a bovine pancreas extract. When 1 ppt of the partially purified substance was added to 2.5% fetal calf serum, the growth of cells was so greatly promoted that it was more than equal to that in the standard medium containing 10% of the serum.     

心包液的活性物质研究进展

心脏及出入心的大血管由内、外两层心包膜所包裹,心包液是位于心包腔内的少量液体,含有1.7%～3.5%的蛋白质,总的蛋白含量低于血浆。近年来研究发现许多心脏病患者的心包液中存在心房利钠素（ANP）、脑钠素（BNP）、内皮素（ET）及金属蛋白酶等活性物质,浓度显著高于血清,而且其水平变化和心功能密切相关。本文就心包液的组成成分及其中的活性物质和心脏疾病及心功能的关系进行了综述。     

Studies on Biologically Active Substances Formed by Bacilli

The products of several Bacillus strains were investigated on rabbit serum calcium decreasing, oxytocic and toad heart function promoting activities. These products were obtained from the clear supernatant fluid of the culture medium after the cells were removed by centrifugation.

For the production of rabbit serum calcium decreasing substance, Bacillus subtilis K and Bacillus natto No. 8 were found to be usefull, Bacillus megaterium KM, Bacillus cereus var. mycoides and Bacillus subtilis K produced oxytocic principle. Bacillus subtilis K, Bacillus brevis and Bacillus megaterium KM also produced toad heart function promoting factor.

A procedure was developed to obtain the electrophoretically homogenous rabbit serum calcium decreasing substance from culture filtrate of Bacillus subtilis K. The crude substance was obtained as isoelectric precipitate by adjusting the culture filtrate to pH 3.0. The crude substance was purified by gel filtration on a Sephadex G-75 column, isoelectric fractionation and chromatography on DEAE-cellulose column. The purified preparation was shown to be homogenous by Tiselius electrophoresis but was separated into two bands by polyacrylamide electrophoresis. The chemical analysis of this biologically active substance indicated this substance to be a lipoprotein. The substance decreased rabbit serum calcium level about 12% at 6~8hr after intravenous injection (dose; 0.5 mg/kg body weight).     

Studies on Substances Active on the Behavior of Planarian

Substances which acted on the behavior of planarian were searched for, and two active components were isolated from culture broth of Streptomyces sp. 340. They were identified as trans-3-methylthioacrylic acid (MTAA) and 3-methylthiopropionic acid (MTPA).

It was found that many Streptomyces and some fungi accumulated MTAA, and many Streptomyces, fungi, bacteria and yeasts accumulated MTPA when the microorganisms were grown in the medium containing methionine.     

Antifeeding Active Substances for Insect in Caryopteris divaricata Maxim.

Six antifeeding active diterpenes having a clerodane skeleton, clerodin (I), caryoptin (II), dihydroc?erodin-I (V), dihydrocaryoptin (VI), clerodin hemiacetal (VII), and caryoptin hemi- acetal (VIII), were isolated from Caryopteris divaricata Maxim. Antifeeding activities of six compounds and these derivatives against the 3rd instar larvae of Spodoptera litura F. were tested by the leaf disk method. In addition, the terms of “relative antifeedant” and “absolute antifeedant” were proposed for the antifeeding substances, and the latter term was used to the above six diterpenes.     

Combination of MS Protein Identification and Bioassay of Chromatographic Fractions to Identify Biologically Active Substances from Complex Protein Sources

Purification of biologically active proteins from complex biological sources is a difficult task, usually requiring large amounts of sample and many separation steps. We found an active substance in a serum response element-dependent luciferase reporter gene bioassay in interstitial cystitis urine that we attempted to purify with column chromatography and the bioassay. With anion-exchange Mono Q and C₄ reversed-phase columns, apparently sharp active peaks were obtained. However, more than 20 kinds of proteins were identified from the active fractions with MS, indicating that the purification was not complete. As further purification was difficult, we chose a candidate molecule by means of studying the correlation between MS protein identification scores and bioassay responses of chromatographic fractions near the active peaks. As a result, epidermal growth factor (EGF) was nominated as a candidate molecule among the identified proteins because the elution profile of EGF was consistent with that of the bioassay, and the correlation coefficient of EGF between MS protein identification scores and bioassay responses was the highest among all the identified proteins. With recombinant EGF and anti-EGF and anti-EGF receptor antibodies, EGF was confirmed to be the desired substance in interstitial cystitis urine. This approach required only 20 ml of urine sample and two column chromatographic steps. The combination of MS protein identification and bioassay of chromatographic fractions may be useful for identifying biologically active substances from complex protein sources.Purification and identification of biologically active proteins existing in minute amounts from biological sources such as urine is still a difficult task (1). It requires a large volume of the sample and many separation steps for purification (2, 3). Nevertheless the recent progress of MS has dramatically changed protein analysis (4). With MS, smaller protein samples can be used than with classical protein identification methods such as N-terminal peptide sequencing.Interstitial cystitis (IC)¹ is a chronic inflammatory disease characterized by frequency and urgency and/or severe pelvic pain (5). The International Continence Society also selected the term “painful bladder syndrome” for IC (6). The quality of life of IC patients is extremely low because of their severe symptoms. The pathogenesis of IC is unclear, and effective treatments have not been established. To elucidate the mechanism of IC pathogenesis, we attempted to find characteristic proteins in IC urine using proteomics techniques and have already reported active neutrophil elastase as an IC urinary marker (7). We had also performed gene expression analysis of IC bladder tissues using GeneChip technology and found that mRNA expression of GPR18, a member of the G-protein-coupled receptors, was higher in IC bladder than in the control.² We tried to confirm whether GPR18 endogenous ligand existed in IC urine by using a bioassay with GPR18 transfectant cells.In the present study, the existence of an active substance in IC urine was suggested in the bioassay using the serum response element (SRE)-dependent luciferase reporter gene with the stable recombinant HEK293 cell line expressing GPR18. We thought that the response was derived from GPR18 and tried to purify the active substance from a small volume of IC urine using chromatographic techniques. Among the many proteins identified from partially purified samples, we clearly nominated epidermal growth factor (EGF) as a candidate molecule judging from the correlation between MS protein identification and the bioassay of chromatographic fractions. With recombinant EGF and anti-EGF antibody, EGF was confirmed to be the desired substance found in IC urine. The complete inhibition of the bioassay response by anti-EGF receptor antibody also indicated that the response was based on the EGF receptor, not GPR18, suggesting that GPR18 overexpression enhanced the EGF signal via the endogenous EGF receptor of the HEK293 cell line.     

大蒜中生理活性物质对致病细菌的体外抑制作用

目的：从鲜蒜中提取蒜氨酸、蒜氨酸酶和蒜素,测定了蒜氨酸、蒜氨酸酶、蒜氨酸＋蒜氨酸酶、蒜素和蒜汁对10种致病细菌的抑菌作用及最低抑菌浓度。方法：采用平板打孔法和液体2倍稀释法。结果：蒜氨酸酶、蒜氨酸＋蒜氨酸酶、蒜素和蒜汁对金黄色葡萄球菌、大肠杆菌、鲍曼不动杆菌等10种菌株的抑菌环直径均大于7mm。蒜氨酸＋蒜氨酸酶对鲍曼不动杆菌的抑菌作用是50mg／mL氨苄青霉素的1．5倍。蒜氨酸＋蒜氨酸酶对金黄色葡萄球菌、类产碱假单胞菌和福氏志贺氏菌的MIC为0．19mg/mL。结论：蒜氨酸酶、蒜氨酸＋蒜氨酸酶、蒜素和蒜汁在体外具有明显的抑菌作用。     

马尾松叶生化物质的生物检测与物质鉴定

马尾松是我国特有的乡土树种,长期林业生产实践表明马尾松纯林连栽会引起马尾松生产量下降。本实验采用超临界萃取技术提取马尾松叶生化物质,并采用马尾松种子发芽试验进行生物检测及物质鉴定。生物检测结果表明,纯CO2提取的马尾松叶生化物质以及乙醇和CO2混和提取的马尾松叶生化物质对马尾松种子的发芽具有不同的效应。总体效果而言,纯CO2提取的马尾松叶生化物质对马尾松种子发芽率、胚根长、鲜重和干重具有不同程度的抑制效应,而对发芽势及胚轴长则具有促进效应;乙醇和CO2混和提取的马尾松叶生化物质对马尾松种子发芽率、胚轴长、鲜重和干重具有不同程度的抑制作用,而对发芽势及胚根长则具有促进作用。物质鉴定结果表明纯CO2提取的马尾松叶生化物质主要是苯甲醛、Cis-2,6-二甲基-2,6-辛二烯等,乙醇和CO2混和提取的马尾松叶生化物质主要是β-芳樟醇、苯乙醇等。     

Purification and Identification of the Fruiting-Inducing Substances in Coprinus macrorhizus

Substances which are effective in inducing fruiting bodies in monokaryotic mycelia of the fis(+) strain of Coprinus macrorhizus were purified and characterized. The active components of fruiting-inducing substances were identified as adenosine-3'-monophosphate, adenosine 3',5'-cyclic monophosphate (cyclic AMP), and a protein which is bound with the cyclic AMP. Cyclic AMP was synthesized from adenine within mycelia of the mutant strains which form monokaryotic fruiting bodies without the addition of fruiting-inducing substances, but not in those of the strains which do not form monokaryotic fruiting bodies. The proteins which bind with cyclic AMP were detected in crude extracts of mycelia of those strains which form monokaryotic fruiting bodies and of the dikaryon, but not in those of the strains which do not form monokaryotic fruiting bodies.     

来源于解淀粉芽胞杆菌的芸薹根肿菌抑菌活性物质的分离与鉴定

十字花科根肿病是由芸薹根肿菌引起的较为严重的世界性病害之一,至今还无有效控制该病害的生物农药。本研究通过盆栽实验,筛选到一株解淀粉芽胞杆菌HB_26,对根肿病原菌具有60%以上的抑制活性。通过高效液相质谱(LC_MS)对活性物质进行分离纯化,根据分子量和紫外吸收光谱初步鉴定活性物质为肽类,命名为BA30。抑真菌实验证明BA30在150μg·mL-1的浓度下,对小麦赤霉和灰霉病菌有70%的抑菌活性,对蚕豆锈病和水稻纹枯病菌有50%的抑菌活性,对番茄早疫病菌有30%的抑菌活性。     

Antioxidative and Prooxidative Abilities of some Biological Substances and Physiologically Active Substances on the Oxidation of Unsaturated Fatty Acids

Some biological substances and physiologically active substances such as amino acids, hormones, vitamins, medicines and the other substances were added into the linoleic acid solution; and antioxidative and prooxidative abilities of these substances, the abilities to act as electron donors or acceptors, were tested. Many substances were effective as the antioxidant and some others showed prooxidative effects. These facts suggested the possibility of the occurrence of many reactions involving the transfer of an electron among biological processes.     

几株产活性物质的海洋细菌的分离与初步鉴定

从海泥、海水及海洋动物中分离到数十株细菌 ,对其产生活性物质的菌株进一步筛选 ,得到产蛋白酶、淀粉酶和抑菌活性等生物活性物质的几株海洋细菌 ,将其中的活性高的四株菌纯化后进行菌种鉴定。通过对革兰氏染色、个体及群体形态观察、糖类发酵、硝酸盐还原等生理特性的研究 ,依据《伯杰氏细菌鉴定手册》确定它们的分类地位 ,它们分别应归属为欧文氏菌属 (Erwiniasp .)、气单孢菌属 (Aeromonassp .)、微球菌属 ( (Micrococoussp .)、和假单孢菌属 ( (Pseudomonassp .)。     

Identification of an Ectokinase Activity in Cerebellar Granule Primary Neuronal Cultures

Abstract: Primary neuronal cultures from 8-day-old rat cerebellum were incubated in the presence of exogenously added 16 n M [γ-³²P]ATP. Phosphorylation of a 45-kDa endogenous protein was detected within 1 min and increased linearly for ∼20 min. Unlike what was seen with [γ-³²P]ATP, in the presence of [³²P] orthophosphate no visible phosphorylation of protein was detected after 10 min, but a different pattern of phosphorylation was obtained in 30 min. The phosphorylation of the 45-kDa protein was reduced by 80–90% in the presence of 1 µ M unlabeled ATP, 5 U/ml of apyrase, or 0.01% trypsin but not 1 m M PO₄³⁻. Phosphorylation was inversely proportional to cell density and was unaffected by addition to the cells of 56 m M KCl or 100 µ M glutamate for 3 min. The presence of exogenously added cellular protein extracts or pretreatment of the cells for up to 20 min in phosphorylation buffer also did not affect the observed phosphorylation of the 45-kDa protein. The phosphorylation was found to be insensitive to MgCl₂ but inhibited in the presence of MnCl₂ or NaF and in the absence of CaCl₂. Analogues of ATP suppressed phosphorylation of the 45-kDa protein by 80–90%. A similar inhibition was obtained in the presence of ADP or AMP. In this study, we establish via several different means that the phosphorylation of the 45-kDa protein in primary neuronal granule cultures occurs extracellularly through an ectokinase activity, which is furthermore distinguishable from a series of other presently characterized ecto-protein enzymes and intracellular kinases.