Anti-Toxoplasma activity of vegetal extracts used in West African traditional medicine

Both Toxoplasma gondii and Plasmodium are Apicomplexan protozoa that share common metabolic pathways and potential drug targets. The objective of this study was to examine the anti-Toxoplasma activity of nine West African plants with known activity against P. falciparum. The extracts were obtained from parts of plant commonly used, by most traditional healers, in the form of infusion or as water decoction. The in vitro activity of plant extracts on T. gondii was assessed on MRC5 tissue cultures and was quantified by enzyme-linked immunoassay. Aqueous extracts from Vernonia colorata were found to be inhibitory for Toxoplasma growth at concentrations > 10 mg/L, with an IC50 of 16.3 mg/L. A ten-fold gain in activity was obtained when organic solvents such as dichloromethane, acetone or ethanol were used to extract V. colorata's active principles. These extracts were inhibitory at concentrations as low as 1 mg/L, with IC50 of 1.7, 2.6 and 2.9 mg/L for dichloromethane, acetone and ethanol extracts respectively. These results indicate a promising source of new anti-Toxoplasma drugs from V. colorata and African medicinal plants.     

苍耳等48种植物提取物的杀虫活性

采用浸虫法测定了48种植物的丙酮提取物对小菜蛾（Plutella xylostella L．）的杀虫活性,并研究了苍耳（Xanthium sibiricum Patrin．）不同溶剂提取物对小菜蛾和棉蚜（Aphis gossypii Glover）的触杀作用以及苍耳丙酮提取物不同溶剂萃取物对小菜蛾的触杀作用。结果表明,在供试的48种植物中,500g·L^-1黄花蒿（Artemisia annua L）、泽漆（Euphorbia helioscopia L．）、夏至草[Lagopsis supina（Steph．ex Willd．）Ik．-Gal．ex Knorr．]、苍耳及律草[Humulus scandens（Lour．）Merr．]的丙酮提取物对小菜蛾的杀虫活性较强。在苍耳的不同溶剂提取物中,苍耳丙酮提取物的杀虫活性最强,乙醇和乙酸乙酯提取物的杀虫活性次之。另外,50g·L^-1苍耳丙酮提取物的氯仿萃取物对小菜蛾的杀虫活性最强,24h和48h的校正死亡率分别为88．33％和91．67％。     

Activity of Brazilian and Bulgarian propolis against different species of Leishmania

Extracts of propolis samples collected in Brazil and Bulgaria were assayed against four Leishmania species--Leishmania amazonensis, L. braziliensis, L. chagasi from the New World, and L. major from the Old World--associated to different clinical forms of leishmaniasis. The composition of the extracts has been previously characterized by high temperature high resolution gas chromatography coupled to mass spectrometry. Considering the chemical differences among the extracts and the behavior of the parasites, it was observed significant differences in the leishmanicidal activities with IC50/1 day values in the range of 2.8 to 229.3 microg/ml . An overall analysis showed that for all the species evaluated, Bulgarian extracts were more active than the ethanol Brazilian extract. As the assayed propolis extracts have their chemical composition determined it merits further investigation the effect of individual components or their combinations on each Leishmania species.     

Demonstration of immunochemical identity between the nerve growth factor-inducible large external (NILE) glycoprotein and the cell adhesion molecule L1.

The nerve growth factor-inducible large external (NILE) glycoprotein and the neural cell adhesion molecule L1 were shown to be immunochemically identical. Immunoprecipitation with L1 and NILE antibodies of [3H]fucose-labeled material from culture supernatants and detergent extracts of NGF-treated rat PC12 pheochromocytoma cells yielded comigrating bands by SDS-PAGE. NILE antibodies reacted with immunopurified L1 antigen, but not with N-CAM and other L2 epitope-bearing glycoproteins from adult mouse brain. Finally, by sequential immunoprecipitation from detergent extracts of [35S]methionine-labeled early post-natal cerebellar cell cultures or [3H]fucose-labeled NGF-treated PC12 cells, all immunoreactivity for NILE antibody could be removed by pre-clearing with L1 antibody and vice versa.     

Evaluation of mutagenic activity in supply water at three sites in the state of Rio Grande do Sul, Brazil

In this study, the Salmonella/microsome assay, using the micro-suspension method, was utilized to evaluate water for public supply at three sites in Rio Grande do Sul. The first site selected was in an area under industrial influence and the others were in non-industrial reference areas. Based on 40 L samples of raw water and/or after conventional treatment, compounds were extracted with XAD4 resins using natural and acidic pH, and the extracts were analyzed in the TA98 and TA100 strains with and without S9. Raw water extracts in the industrial region induced 27.4 revertants/L (rev/L) for TA100+S9 up to 226.3 rev/L for TA100-S9, both for acidic pH extracts. After conventional treatment the responses varied from 20.6 rev/L (TA98-S9) for natural pH extracts to 755.5 rev/L (TA98-S9) for acidic pH extracts. For acidic extracts obtained from reference site samples, the response, with metabolic activation only, ranged from negative to minimal. Direct mutagenic responses in acidic extracts may be elevated in treated extracts, compared with raw water extracts, influenced by the presence of by-products of the chlorination process. However, the mutagenicity observed in the treated water extracts in the industrial area increased and reflected mainly the combination of directly and indirectly acting compounds in the source waters, that are heavily influenced by anthropogenic factors.     

Evaluation of in vitro antifungal activity of medicinal plants against phytopathogenic fungi

Fourteen medicinal plants belonging to 13 families were collected and extracted with petroleum ether (PE), chloroform, methanol and water to yield 60 crude extracts. Using agar diffusion method, these extracts were evaluated for antifungal activity on the growth of five phytopathogenic fungi. Among all the extracts tested, PE, chloroform and methanol extracts of Piper betle L. and PE and chloroform extracts of Allamanda cathartica exhibited promising antifungal activity. Minimum inhibitory concentration (MIC) values of the above promising extracts were determined using broth dilution technique and observed that chloroform extract of P. betle L. exhibited the least MIC value ranging from 280 to 1130 μg ml^?1. In this study, we report chloroform extract of P. betle L. to be thermally stable even when steam sterilised for the first time and that it could be stored at 4°C with almost no change in its activity for a period of 180 days.     

6种陆生植物提取物对白脊藤壶无节幼体的毒杀活性比较

对夹竹桃科（Apocynaceae）的红花夹竹桃（Nerium indicum Mill．）和黄花夹竹桃（Thevetia peruviana K．Schum．）、百合科（Li｜iaceae）的洋葱（Allium cepa L．）和大蒜（A．sativum L．）、楝科（Meliaceae）的苦楝（Melia azedarach L．）和印楝（Azadirachta indica A．Juss．）等6种陆生植物的不同溶剂提取物对白脊藤壶（Balanus albicostatus Pilsbry）无节幼体的毒杀作用进行了比较研究。结果表明,供试的楝科和夹竹桃科的4种植物提取物对白脊藤壶无节幼体的毒杀活性高于洋葱和大蒜;不同提取物的毒杀活性与植物的种类及提取部位（叶、花）以及活性成分的类型有关,用非极性有机溶剂提取的脂溶性提取物的毒杀活性高于极性溶剂提取物。处理12h,0．3％印楝素乳油的乙酸乙酯及95％乙醇稀释液的毒杀活性最高,LC50仅为0．33和3．01g·mL^-1,可作为防治海洋污损生物的植物资源进行进一步的开发利用。     

落叶松水浸液对胡桃楸种子萌发和幼苗生长的影响

研究了落叶松水浸液对胡桃楸种子萌发和幼苗生长的影响.结果表明,与对照相比,25 g ·kg^-1浓度的落叶松根水浸液,125 g·kg^-1浓度的落叶松皮、枝水浸液对胡桃楸种子发芽率、发芽指数均表现出显著的促进作用（P＜0.05）;12.5 g·kg^-1和25 g·kg^-1浓度的落叶松根水浸液、12.5 g·kg^-1浓度的落叶松皮水浸液对胡桃楸幼苗苗高、地径生长的促进作用显著（P＜0.05）.高浓度的落叶松叶水浸液对胡桃楸种子萌发和幼苗生长具有明显的促进作用（P＜0.05）.而落叶松叶水浸液对胡桃楸种子发芽率、发芽指数、苗高以及地径的促进作用却随着水浸液浓度的降低,呈逐渐下降的趋势.施加落叶松水浸液对胡桃楸种子萌发以及幼苗生长的促进作用可能与树种间的化学相互作用有关.     

Effects of two plant extracts on larval leafminer Liriomyza trifolii (Diptera: Agromyzidae) in tomatoes

Aqueous extracts from two plants, Urginea maritima L. (Liliaceae) and Euphorbia myrsinites L. (Euphorbiaceae), were tested for their insecticidal activity against the leafminer Liriomyza trifolii (Burgess) on infested tomato, Lycopersicon esculentum Mill., plants in the laboratory and field. Two grams of plant material was extracted with 100 ml of water and then diluted 1:100, 1:50, and 1:25 with distilled water. Diluted plant extract was either applied to the infested tomato leaves or by soil drench and was compared with foliar application of cyromazine. All dilutions of both plant extracts caused significant control of the leafminer larvae and maintained populations below those of the nontreated control plants in all trials. Only at the most concentrated dilutions (1:25) were the plant extracts statistically similar to the cyromazine treatment. Furthermore, greenhouse yields from all of the foliar treatments were statistically similar to the cyromazine treatment and significantly better than the nontreated control. Four species of leafminer parasitoids were found in the greenhouse; however, the percentage of parasitism was significantly less in all treated replicates than in the nontreated control replicates. Aqueous extracts from these two plant extracts exhibited both translaminar and systemic activity and are potential candidates as new organic insecticides.     

Modified nitrocefin‐EDTA method to differentially quantify the induced L1 and L2 β‐lactamases in Stenotrophomonas maltophilia

Aim: To estimate the ethylene diamine tetraacetic acid (EDTA) concentration at which the L1 enzyme activity in the cell extracts of Stenotrophomonas maltophilia can be mostly inhibited. Methods and Results: The effective inhibition concentration of EDTA against the L1 enzyme in the cell extracts was firstly evaluated by using the L2 isogenic mutant of S. maltophilia KJ, KJΔL2, as the assayed strain. Approximately 92% L1 activity was inhibited by 10 mmol l^?1 EDTA, which is 100‐fold higher than that from previously reported protocols (0·1 mmol l^?1). Three phylogenetic clusters of L1 proteins were revealed from 11 clinical S. maltophilia isolates, with a L1 protein divergence of 0–11%. The EDTA concentration required to inhibit the L1 enzymes of different phylogenetic clusters was estimated to be 10 mmol l^?1. Conclusion: The previous nitrocefin‐EDTA protocol for differentially quantifying the L1 and L2 activity in the cell extracts has been modified by raising the added EDTA concentration to 10 mmol l^?1. Significance and Impact of the Study: A rapid and accurate method for determination of L1 and L2 activity will provide a convenient tool for enzyme characterization and induction mechanism study of S. maltophilia.     

Alterations in Endogenous Levels of Gibberellin-like Substances during Germination of Phaseolus vulgaris Seeds

The endogenous gibberellin-like substances were determined in mature dry and germinating bean seeds, Phaseolus vulgaris L. cv. Alabaster. Methanol extracts were partitioned against ethyl acetate and butanol at neutral and acid pH. Each phase was individually chromatographed on a silica gel column. The gibberellin activity was measured with the Tan-ginbozu dwarf rice micro-drop bioassay. Each extract was tested in two dilutions. Extracts from dry seeds showed the highest gibberellin activity, largely attributable to ethyl acetate-soluble substances. The activity was considerably reduced in extracts from seeds imbibed for 1 day. Gibberellin-like substances soluble in butanol appeared in extracts from seeds soaked for 1 or more days.     

Toxicity of Leaf and Stem Extracts to Tylenchorhynchus dubius

Plant extracts, made by grinding 2 g of fresh tissue in 5 ml of water, were toxic to Tylenchorhynchus dubius and Hoplolaimus spp. Such extracts from leaves and stems of bean (Phaseolus vulgaris L.) and leaves of tobacco (Nicotiana tabacum L.) were most toxic; those from leaves of corn (Zea mays L.), tomato (Lycopersicon esculentum Mill.) and rhododendron (Rhododendron catawbiense L.) were less toxic; and extracts of bean roots were nontoxic. Nematode movement slowed markedly within 1 hr in tobacco leaf extract, and within 4 hr in bean leaf extract; both extracts completely inactivated or killed 95% of the nematodes in 24 hr. Heating leaf extract 10 min at 80 C eliminated toxicity. Absorption of fusicoccin, a phytotoxin produced by Fusicoccum amygdali Del., increased the toxicity of tomato leaf extracts, whereas water extracts of acetone-extracted powder preparations of leaves were about 15-fold more toxic than water extracts of fresh tissue. Addition of homogenized leaves of bean, tobacco and tomato to soil significantly reduced nematode populations within 3 days.     

灰毛豆甲醇提取物的杀虫活性

研究灰毛豆(Tephrosia purpurea)各部位提取物的杀虫活性及其作用方式。结果表明,灰毛豆对白纹伊蚊Aedes albopictus Skuse幼虫、菜青虫Pieris rapae(L.)、斜纹夜蛾Prodenialitura(Fabricius)幼虫和黄曲条跳甲Phyllotreta striolata(Fabricius)成虫都有杀虫活性。灰毛豆种子、树皮、根皮、豆荚、枝条和树叶的甲醇提取物对白纹伊蚊4龄幼虫24h的LC50值分别是22.1,97.7,36.6,142.6,165.6和618.3mg.L-1,树干木质部没有杀虫活性;灰毛豆种子、树皮和根皮甲醇提取物对3龄菜青虫24h的触杀毒力LC50值分别是232.1,206.3和236.7mg.L-1;种子、树皮、根皮、枝条、树叶和豆荚对3龄菜青虫的24h胃毒毒力LC50值分别是192.6,168.4,249.7,524.5,1001.0,和510.7mg.L-1。在取食或接触有提取物的叶碟后,5龄菜青虫会出现取食量减少和生长发育变慢的亚致死现象。这些研究结果表明,灰毛豆除茎干木质部以外的其它各部位均含有杀虫活性成分,其作用方式为胃毒和触杀。     

Differentiation of Legionella species by soluble protein patterns in polyacrylamide slab gels

Water-soluble protein extracts were obtained from representative strains of eleven species of Legionella and examined by polyacrylamide slab gel electrophoresis (PAGE). The eleven species included: L. pneumophila (serogroups 1-9), L. micdadei, L. longbeachae (serogroups 1 and 2), L. gormanii, L. dumoffii, L. bozemanii, L. jordanis, L. oakridgensis, L. wadsworthii, L. feeleii and L. maceachernii. Species identification was verified by gas-liquid chromatography of the non-hydroxy, monohydroxy and dihydroxy fatty acids. Strains within each species shared common patterns of protein banding. Each species had a distinctive protein banding pattern. Results indicate that PAGE of water soluble protein extracts is a useful technique for identification of Legionella isolates.     

白桦、落叶松不同器官水浸液对种子萌发和播种苗生长的种间化感作用

研究了白桦的根、枝、叶,落叶松的根、枝、叶和皮不同浓度水浸液(5.0、12.5、25.0、50.0和100.0 mg·mL-1)对另一树种种子萌发和当年生播种苗生长的化感作用.结果表明:落叶松不同器官水浸液(根5.0 mg·mL-1除外)处理均抑制白桦种子的萌发,其平均发芽率为:对照(75％)＞根(66％)＞皮(59％)＞枝(58％)＞叶(54％),随浓度增加,根、枝水浸液的抑制作用增强,而叶、皮水浸液的抑制作用减弱.落叶松各器官水浸液对白桦胚根和胚轴生长具有较强的抑制作用,且以100.0 mg·mL-1浓度的叶水浸液抑制作用最强,胚根和胚轴长分别降低38％和55％(P＜0.05).落叶松枝、叶水浸液处理对白桦播种苗的苗高、地径及生物量有一定的促进作用,根、皮水浸液则具有抑制作用.白桦各器官水浸液能促进落叶松种子的萌发,根和枝的水浸液促进胚轴生长,叶的水浸液抑制胚轴生长,50.0和100.0 mg·mL-1浓度的叶水浸液处理的种子胚轴分别较对照降低27％和28％(P＜0.05).其对落叶松播种苗生长的影响以促进作用为主,5.0 mg·mL-1浓度的白桦叶水浸液处理的落叶松苗高、地径、生物量分别较对照高54％、60％和100％ (P＜0.05).白桦与落叶松之间存在明显的化感作用,混交可能对林木生长产生一定的促进作用.     

Serine Proteinase Inhibitors from Eggs and Larvae of Tick Boophilus microplus: Purification and Biochemical Characterization

The present study describes the purification, characterization, and comparison of serine proteinase inhibitors during the development of egg and larva phases of the tick Boophilus microplus. Samples were collected of eggs between the first day of hatching and the beginning of eclosion (defined as E1, E2, and E3) and of larvae between the first day of eclosion and the infectant phase (defined as L1, L2, and L3). Crude extracts of the samples (2.5% w/v in Tris-HCl buffer) were analyzed by SDS-PAGE, and showed three major protein bands of 42, 62, and 85 kDa, differing in intensity, from E1 to L3 samples. The total protein of the larva extracts was 34% less than that of the egg extracts, while no differences in active protein were detected. The apparent dissociation constant K _i determined for trypsin was 10-fold lower from E1 to L3 samples. Serine proteinase inhibitors from tick eggs and larvae (BmTIs) were purified on trypsin-Sepharose column and analyzed by SDS-PAGE. The results showed a slight difference in protein pattern, with a protein band of 20 kDa in the E1 and E2 samples which did not appear in the other samples. The K _i for neutrophil elastase was 10-fold lower in L3 than E1. BmTI reverse-phase chromatography showed two and one major peaks in egg and larva samples, respectively. The N-terminal amino acid sequence of the L3 main peak from a C8 column showed a mix of BmTIs with the major sequence AVDFDKGCVPTADPGPCKG. Changes indicated by molecular weight and inhibition activity suggest different roles for BmTIs during the development process.     

Evidence for the synthesis of thymosin alpha 1 by calf thymocytes and the production of this peptide by natural processing

Thymus and thymocytes from calf were extracted under isotonic conditions in the presence of protease inhibitors or under severe denaturing conditions (after quick freezing and thawing in boiling 0.1 M NaCl). The extracts, as well as the medium in which the thymocytes were obtained from thymus fragments (thymocyte supernatants), were size-fractionated by ultrafiltration. As in whole thymus isotonic extracts, thymosin alpha 1 [A. L. Goldstein, T. L. K. Low, M. McAdoo, J. McClure, G. B. Thurman, J. Rossio, C-Y. Lai, D. Chang, S-S. Wang, C. Harvey, A. H. Ramel, and J. Meienhofer (1977) Proc. Natl. Acad. Sci. USA 74, 725-729] was contained in isotonic extracts from thymocytes and also in thymocyte supernatants, as determined by isoelectric focusing and reverse-phase HPLC analysis. The extraction under denaturing conditions mainly yielded products with molecular masses over 50,000, showing very similar isoelectric focusing patterns in both thymocytes and whole thymus extracts. As deduced by isoelectric focusing analysis of diverse size-fractionated products, a strong association capacity seems to be responsible for an apparently high molecular mass of the components of these extracts. According to the pI, two of these components were prothymosin alpha [A. A. Haritos, G. J. Goodall, and B. L. Horecker (1984) Proc. Natl. Acad. Sci. USA 81, 1008-1011] and thymosin alpha 1. Prothymosin alpha was not detected in any isotonic extracts or thymocyte supernatants. These data suggest that calf thymocytes are capable of producing thymosin alpha 1, which would arise by natural processing of its precursor.     

Screening of certain medicinal plants from India for their anti-quorum sensing activity

Discovery of quorum sensing (QS) system to coordinate virulence and biofilm formation in bacterial pathogens has triggered search for safe, stable and non-toxic anti-QS compounds from natural products. Ethanolic extracts of 24 Indian medicinal plants were tested by agar well and disc diffusion assay for anti-QS activity using Chromobacterium violaceum (CV12472 and CVO26) reporter strains. AHL from C. violaceum CV31532 was isolated and partially purified for its use in CVO26 based bioassay. Effect on swarming-motility of Pseudomonas aeruginosa (PAO1) was also recorded at sub-MIC concentrations of extracts. Of the 24 medicinal plants screened Hemidesmus indicus (L.) Schult (root), Holarrhena antidysenterica (Roth) A.DC. (bark), Mangifera indica L. (seed) Punica granatum L. (pericarp) and Psoralea corylifolia L. (seed) demonstrated varying level of inhibition of violacein production in the reporter strains. Moreover, a significant reduction in swarms was recorded over control. The inhibition of violacein production and swarming motility may be due to direct or indirect interference on QS by active constituents or the interactive effect of different phytocompounds present in the extracts. These plant extracts may be selected for activity guided fractionation to identify and characterize the active principle.     

Evidence of an antilisterial factor induced by wounding of iceberg lettuce tissues

AIMS: To examine the influence of wound-associated reactions in cut iceberg lettuce (Lactuca sativa L.) tissues on the fate of Listeria monocytogenes. METHODS AND RESULTS: Aqueous extracts prepared from shredded iceberg lettuce before and after storage in high oxygen permeability film were inoculated with L. monocytogenes. Listeria monocytogenes grew in extracts prepared from fresh lettuce. In contrast, inhibition ranging from arrested growth to a decline in cell viability was observed in extracts prepared from samples stored for 1-3 days. Similar behaviour was evident in lettuce shreds inoculated with 10(5) CFU g(-1)L. monocytogenes immediately after processing or after 3 days in storage. Heat treatment of the cut tissues at 47 degrees C for 3 min before storage diminished the inhibitory effect. CONCLUSIONS: The results provided evidence that an antilisterial factor or factors are released by wounded iceberg lettuce tissues. Antilisterial activity was mitigated by heat treatment of the lettuce. SIGNIFICANCE AND IMPACT OF STUDY: This study indicates that intrinsic factors associated with plant metabolism could play a significant role in the ecology of human pathogens in packaged horticultural products.     

泥胡菜等8种草本植物提取物除草活性的生物测定

采用顺序提取法制备了泥胡菜（Hemistepta lyrata Bunge）等8种草本植物全草的石油醚、乙酸乙酯和乙醇提取物,并以高粱（Sorghum vulgare Pers．）、黄瓜（Cucumis sativus L．）、小麦（Triticum aestivum L．）和油菜（Brassica campestris L．）为供试对象,用种子萌发法对提取物的除草活性进行了生物测定。结果表明,所有提取物对4种作物幼苗根和茎的生长均有一定的抑制作用,但抑制率有一定差异。泥胡菜和獯草[Humulus scandens（Lour．）Merr．]的乙酸乙酯提取物对作物幼苗根和茎的抑制作用最强,抑制率随提取物浓度的提高逐渐增高,且对作物幼苗根生长的抑制强度高于茎。在低浓度（12．5g·L^-1）条件下,泥胡菜乙酸乙酯提取物对小麦幼苗根和茎生长的抑制作用最强;蓓草乙酸乙酯提取物对高粱幼苗根的生长及油菜幼苗根和茎的生长抑制作用最强。研究结果显示,泥胡菜及蓰草的乙酸乙酯提取物具有潜在的除草活性。