Anti-inflammatory activity of shikonin derivatives from Arnebia hispidissima.

Arnebia hispidissima ethanolic extract, after chromatography, yielded a number of shikonin derivatives, which were identified as arnebin-5, arnebin-6, teracryl shikonin, arnebinone and acetyl shikonin. All these compounds were firstly reported from this plant species and evaluated to the anti-inflammatory activity of ethanolic extract and isolated shikonin derivatives, models with carrageenan-induced paw edema and complete Freund's adjuvant (CFA)-induced chronic arthritis in rats were conducted. The observed results indicated that pre-treatment with arnebinone significantly inhibited the carrageenan-induced paw edema and also suppressed the development of chronic arthritis induced by CFA.     

Cytoplasmic Channels and Chromatin Migration in the Meiocytes of Arnebia hispidissima (Sieb.) DC

Microsporogenesis was studied in Arnebia hispidissima whichprovided interesting evidence for the occurrence of cytoplasmicchannels and DNA migration between the meiocytes. The intercommunicatingchannels (0.25–1.75 µ in diameter) are consideredto be a prerequisite for chromatin migration and to act as pathwaysbetween the connected meiocytes. They may develop at any stageof meiosis from early prophase to metaphase II and the migrationmay take place at any time following channel formation. Thetendency of chromatin to migrate seemed to be higher in theearly prophasic stages when it moved in the form of ill-definedmasses or clumps. The chances of migration as well as the amountof migrating chromatin is reduced as the meiosis advances. Theamount of chromatin migrating varies from a small portion tothe entire chromatin mass or nucleus. Pollen mother cells withdouble the normal amount of chromatin as well as those withlittle or none were studied as well as cells with increasedor decreased chromosome numbers. In the chromatin mass, uni-or bivalents stretched between the meiocytes or attenuated towardsthe channel, were frequently seen. Occasional semi-asyndesis,multivalent formation, precocity, and nondisjunction was alsonoted. The possibility of channel formation and the induction of chromatinmovement as a consequence of mechanical or procedural defectis ruled out. It is believed that these cytoplasmic channelshave no relation with the plasmodesmata but they arise de novoand that chromatin migration occurs as a natural phenomenon(in some species). The observations are not due to faulty fixation,staining, or squashing.     

Sphingomyelinase activities in neuronal cell cultures.

Human and rat brains have been previously demonstrated to contain three sphinomyelinases, one lysosomal with a pH optimum of 5.0, one with a pH optimum of 7.4 and a dependence on magnesium and another with a pH optimum of 7.0 and no divalent cation requirement. Using brain cell cultures and clonal cell lines of both neuronal and glial origin the activities of the pH 5.0 and pH 7.4 (magnesium-dependent) sphingomyelinase were examined. Sphingomyelinase activity measured at pH 5.0 was found in all the cell lines tested including G26, C6, N18 (differentiated and undifferentiated), mouse “L” cells, human skin fibroblasts, fetal mouse brain surface cultures and fetal mouse brain reaggregate cultures. However, pH 7.4 (magnesium-dependent) sphingomyelinase activity was found only in the N18 cell lines and the reaggregate cultures suggesting the probable localization of this activity in neurons. Although the pH 7.4 sphingomyelinase activity was found in the fetal mouse brain used for the surface cultures this activity was rapidly lost. This enzyme may play an important role in neuronal development and maturation.     

Synthesis of distinctly different sets of antimicrobial activities by elicited plant cell suspension cultures

The aim of this study was to evaluate the potential of plant suspension cultures for the production of antimicrobial activities. The extracellular, intracellular and cell wall bound fractions of 16 heterotrophic, photoautotrophic and photomixotrophic plant cell suspension cultures each treated with nine different elicitors were tested for the elicitor dependent production of antimicrobial activities. Distinctly different patterns of bioactivities directed against a panel of human isolates including Gram-positive (Bacillus subtilis, Staphylococcus aureus) and Gram-negative (Escherichia coli, Pseudomonas aeruginosa) bacteria as well as fungi (Candida maltosa) were identified for all except the two autotrophic cell cultures. The intracellular fractions of elicited cell cultures were more active than extracellular fractions while cell wall bound fractions showed almost no activities. The intracellular fraction of heterotrophic Lavendula angustifolia cells elicited with a preparation of Pseudomonas syringae was the most active fraction against Candida maltosa. The intracellular fraction of photomixotrophic Arabidopsis thaliana cells elicited with salicylic acid was active against all test isolates. An antimicrobial protein could be identified and partially purified from this culture. Our findings suggest that elicited plant cell cultures may present a new promising alternative source of antimicrobial proteins.     

An efficient in vitro shoot regeneration from immature inflorescence and ex vitro rooting of Arnebia hispidissima (Lehm). DC. - A red dye (Alkannin) yielding plant

Arnebia hispidissima, which belongs to the family Boraginaceae, is an important medicinal and dye yielding plant. The alkannin, a red dye, are root-specific secondary metabolites of A. hispidissima. Shoots were regenerated from callus derived from immature inflorescence explants obtained from field grown plants. MS medium containing 4.52 μM 2, 4-D and 3.33 μM BAP was found to be most effective for the proliferation of callus, induced on medium containing 4.52 μM 2, 4-D. Maximum number (43.1 ± 0.25) with average length (5.2 ± 0.23) of shoots regenerated when callus was transferred to MS medium supplemented with 1.11 μM BAP, 1.16 μM Kin and 0.57 μM IAA. About 75.5 % of in vitro regenerated shoots were rooted on half-strength MS medium supplemented with 9.84 μM of IBA and 200 mg l⁻¹ of activated charcoal. In comparison to in vitro, higher percent (90.2 %) of shoots were rooted under ex vitro conditions when treated with IBA (0.98 mM) for 5 min. Plantlets rooted in vitro as well as ex vitro were acclimatized successfully under the green house conditions. Ex vitro rooted plants exhibited higher survival percentage (75 %) as compared to in vitro rooted plantlets (60 %). Present study may be applicable in the large-scale root-specific red dye (alkannin) production via root induction under ex vitro condition.     

Expression of differentiated activities in reaggregated brain cell cultures.

Dissociated fetal mouse brain cells are allowed to reassociate in rotation culture to form aggregates. After several weeks these reaggregated brain cell cultures show markedly increased specific activities of monoamine oxidase, lactate dehydrogenase, and the brain-specific protein S-100, while catechol-O-methyltransferase activity increases slightly. Similar changes in these activities are found during mouse brain maturation. The amounts of monoamine oxidase, catechol-O-methyltransferase, and S-100 were also determined in surface cultures of fetal mouse brain cells, as well as glioma and neuroblastoma cell lines. The fetal brain and glial cell cultures possess much higher activities than the cultured neuroblastoma cells. However, lactate dehydrogenase activity was highest in the glioma and lowest in the surface cultures of fetal brain cells.     

藻类活性物质对紫草细胞生长和色素形成的影响

本文研究了螺旋藻、栅列藻和织线藻的水提物对新疆紫草和硬紫草细胞生长和色素形成的作用。结果表明不同种类的藻的提取物对不同紫草细胞作用呈现差异。在生长阶段,对新疆紫草,上述3种藻的低浓度提取物促进生长但作用不大,高浓度的螺旋藻和栅列藻提取物强烈抑制生长;对硬紫草,各种藻提取液的所有浓度处理均有促生长作用。在色素形成阶段,连续用高浓度织线藻提取物处理可以加速新疆紫草色素形成,同时提高色素含量。低浓度的织线藻提取液处理能提高硬紫草色素含量。栅列藻的水提物对两种紫草的色素形成均起抑制作用。螺旋藻水提物的适当浓度可加速两种紫草的色素形成。B_5培养基中加高浓度织线藻水提物,可抑制新疆紫草生长阶段的色素形成。     

Trypanocidal and antimicrobial activities of Moquinia kingii.

A chloroform crude extract (aerial part) and two compounds, apigenin (1) and cynaropicrin (2), isolated from Moquinia kingii were evaluated against Trypanosoma cruzi trypomastigotes in vitro. Antimicrobial activity was also screened using twenty-two strains including gram-positive and gram-negative bacteria and the yeasts Candida albicans and C. tropicalis. The chloroform crude extract, fractions and isolated compounds from M. kingii were active for both activities. The IC50 values for trypanocidal activity obtained for cynaropicrin and apigenin were 93.5 microg/ml and 181 microg/ml, respectively, while the minimum inhibitory concentrations (MICs) varied from 100 microg/ml to 2500 microg/ml, against the strains of bacteria and yeasts evaluated.     

Establishment of anthocyanin-producing cell suspension cultures of Cleome rosea Vahl ex DC. (Capparaceae)

The effects of different levels of Murashige and Skoog (MS) basal medium, 2,4-dichlorophenoxyacetic acid (2,4-D), and sucrose on anthocyanin production and biomass accumulation of cell suspension cultures of Cleome rosea were investigated. Cultures were established in liquid MS medium containing 30 g l⁻¹ sucrose and supplemented with 0.90 μM 2,4-D. Proliferating cell suspension cultures achieved the highest growth capacity, a fourfold increase in biomass accumulation, following subculture at the exponential growth phase, 14–18 days of culture. Moreover, the presence of 2,4-D was essential for anthocyanin production and biomass accumulation. On the other hand, increasing levels of sucrose above 30 g l⁻¹ resulted in a drastic reduction in biomass accumulation. Anthocyanin production was highest in cell suspension cultures grown on half-strength MS medium (1/2 MS), 30 g l⁻¹ sucrose, and 0.45 μM 2,4-D. These cell suspension cultures were mainly composed of small aggregates of spherical cells with similar morphology observed in anthocyanin-producing and non-producing cultures. Moreover, microscopic analysis of anthocyanin-producing cultures showed the presence of mixtures of non-pigmented, low-pigmented, and high-pigmented cells.     

Purification and characterization of tyrosine aminotransferase activities from Anchusa officinalis cell cultures

Three activities of tyrosine aminotransferase (TAT; EC 2.6.1.5), the enzyme which catalyzes the first step of the tyrosine pathway leading to the formation of rosmarinic acid (alpha-O-caffeoyl-3,4-dihydroxyphenyllactic acid), have been extensively purified from cell suspension cultures of Anchusa officinalis L. and subsequently characterized. TAT-1, TAT-2, and TAT-3 differ slightly in native molecular weights (180,000-220,000) and are composed of subunits (4 X 43,000 for TAT-1 and 4 X 56,000 for TAT-2). All three enzymes show a pronounced preference for L-tyrosine over other aromatic amino acids, but TAT-2 and TAT-3 can also effectively utilize L-aspartate or L-glutamate as a substrate. For amino acceptor cosubstrates, either oxaloacetate or alpha-ketoglutarate can be utilized equally well by TAT-1, while the former is the most effective alpha-keto acid for TAT-2 and the latter is the best for TAT-3. All the TAT activities display high pH optima (8.8-9.6), and are inhibited by the tyrosine metabolite 3,4-dihydroxyphenyllactate. TAT-2 and TAT-3 are also inhibited by rosmarinic acid.     

Specific effects on enzyme activities upon ditution of petroselinum hortense cell cultures into water.

Large increases in the specific activities of phenylalanine ammonia-lyase (EC 4.3.1.5) and p-coumarate:CoA ligase (EC 6.2.1.-) occurred within a few hours after dilution of cultured Petroselinum hortense cells into water. No significant changes in the total amount of extractable protein and in the activities of chalcone isomerase (EC 5.5.1.6) and glutamate dehydrogenase (EC 1.4.1.2) were observed under the same conditions. The time course for the change in phenylalanine ammonia-lyase activity included a lag period of 2–3 h, a peak about 13 h after the onset of induction, and a subsequent period of rapid decline. The inducible amount of enzyme activity was greatly dependent upon the degree of dilution of the cells into water. Simultaneous induction by dilution and irradiation of the cells with white, fluorescent light resulted in an increase in the phenylalanine ammonia-lyase level of activity which exceeded that calculated from the sum of the separately induced levels. Consecutive inductions, first by dilution and then 5 h later by irradiation, each required a lag period of 2–3 h. Actinomycin D or cycloheximide were inhibitors of the induction. While the total protein-synthesizing capacity in vitro was not significantly influenced by the dilution of cells, analysis of the labeled products on polyacrylamide gels demonstrated small but significant changes in the radioactivity profiles. The results are consistent with the hypothesis that dilution of the cells into water reduces the concentration of one or more compounds of cellular origin, thereby stimulating the rate of de novo synthesis of a limited number of proteins.     

Plant regeneration from callus cultures of Valeriana wallichii DC.

Petiole expiants of Valeriana wallichii. DC., a threatened medicinal plant, were used for inducing callus. Optimum callus formation was observed on Murashige and Skoogs' (1962) medium supplemented with 3.0 mg/l NAA and 0.25 mg/l Kn. Shoot regeneration was achieved upon transferring the callus to medium containing 1.0 mg/l Kn and 0.25 mg/l NAA. Complete plantlets were obtained on the same medium or upon transfer of the regenerated shoot buds to medium containing 5.0 mg/l Kn and 1.0 mg/l IAA. Nearly a thousand callus regenerated plants were successfully transferred to the field following previously standardized hardening procedures.Abbreviations BAP 6-Benzylaminopurine - 2,4-D 2,4-dichloro phenoxyaceticacid - 2iP 2-isopentenyladenine - IAA indole-3-acetic acid - IBA indole-3-butyric acid - Kn Kinetin - MS Murashige and Skoog's medium (1962) - NAA -napthalene aceticacid - Z Zeatin     

Chemical constituents of Lavatera trimestris L.--antioxidant and antimicrobial activities

Nine phenolic compounds, such as cis-/trans-p-coumaric acid, cis-/trans-p-coumaric acid methyl ester, glucose ester of cis-/trans-p-coumaric acid, caffeic acid methyl ester, kaempferol 7-O-beta-D-glucoside and kaempferol 3-O-beta-D-glucoside, were isolated from Lavatera trimestris flowers by chromatographic techniques and their structures were elucidated by spectral means (NMR). All compounds were tested for their antioxidant activity, while the methanolic extract was tested also for its antimicrobial activity. Also several non-polar constituents have been identified using GC and GC/MS methods. This is the first time that phenolic esters and non-polar constituents were identified in the flowers of L. trimestris L.     

Anti-inflammatory and antimicrobial activities of triterpenoids from Strobilanthes callosus nees.

The anti-inflammatory and antimicrobial activities of the 95% ethanol extract, benzene fraction and isolated triterpenoids of Strobilanthes callosus were investigated. In the carrageenan-induced paw edema inflammation model, the taraxerol showed a high reduction of edema, but the antimicrobial effect observed was lower at the two doses employed. These results confirm the use of this plant in folk medicine as an anti-inflammatory and antimicrobial herbal drug.     

蜘蛛抱蛋属植物内生细菌的多样性及其抗菌活性筛选

【背景】药用植物中蕴含多样性丰富的内生菌资源,这些微生物产生的多种新型物质在制药领域表现出较好的应用前景。【目的】研究蜘蛛抱蛋属(Aspidistra Ker-Gawl.)植物内生细菌的多样性,探索药用植物内生细菌在药用活性产物方面的开发潜力,以期发现具有抗菌活性的次级代谢产物。【方法】对9种13株新鲜的蜘蛛抱蛋植物进行表面消毒,采用5种分离培养基分离内生细菌;根据菌落形态特征排除重复菌株,并测定其16S rRNA基因序列,构建系统进化树分析内生细菌多样性;将菌株分别用2种培养基发酵,使用耻垢分枝杆菌(Mycobacterium smegmatis ATCC 700044)、水稻白叶枯菌(Xanthomonas oryzae PXO99A)、白色念珠菌(Candidaalbicans ATCC 10231)、肺炎雷伯菌(Klebsiella pneumoniae ATCC 700603)和耐药粪肠球菌HH22(Enterococcus faecalis HH22)5种检定菌对分离菌株的发酵液进行抑菌活性筛选。【结果】从植物组织中分离得到了234株内生细菌,根据形态初步排重得到156株植物内生细菌;基于16S rRNA基因序列构建的系统进化树显示它们分属于3门10目22科29属,其中链霉菌属(Streptomyces)、芽孢杆菌属(Bacillus)、微杆菌属(Microbacterium)、类芽孢杆菌属(Paenibacillus)和根瘤菌属(Rhizobium)的菌株广泛地分布在不同种的蜘蛛抱蛋植株中,且占据一定优势;发现可能的潜在新分类单元6个;156株内生细菌中38株菌的发酵液具有抑菌活性,初筛阳性率为23.7%。【结论】蜘蛛抱蛋植物组织中含有种类多样的内生细菌,它们可能是抗菌生物活性次级代谢产物的有效来源。