Allyl-thiosulfinates, the bacteriostatic compounds of garlic against Helicobacter pylori

Allicin and allyl-methyl plus methyl-allyl thiosulfinate from acetonic garlic extracts (AGE) have been isolated by high-performance liquid chromatography. These compounds have shown inhibition of the in vitro growth of Helicobacter pylori (Hp), the bacterium responsible for serious gastric diseases such as ulcers and even gastric cancer. A chromatographic method was optimized and used to isolate these thiosulfinates. The method developed has allowed the isolation of natural thiosulfinates extracted from garlic by organic solvents and is an easy and cheap methodology that avoids complex synthesis and purification procedures. The capacity and effectiveness of isolated natural thiosulfinates have been tested, and this has enabled the identification of the main compounds responsible for the bacteriostatic activity shown by AGE origin of these kinds of organosulfur compounds along with ethanolic garlic extracts (EGE). Additionally, microbiological analyses have suggested that these compounds show a synergic effect on the inhibition of the in vitro growth of Hp. The results described here facilitate the process of obtaining garlic extracts with optimal bacteriostatic properties. The product is obtained in a way that avoids expensive purification methods and will allow the design of live tests with the aim of investigating the potential for the use of these garlic derivatives in the treatment of patients with Hp infections.     

Thermal degradation of allicin in garlic extracts and its implication on the inhibition of the in-vitro growth of Helicobacter pylori

Allicin, the main active principle related to Allium sativum chemistry, is considered to be responsible for the bacteriostatic properties of garlic. The work described here has demonstrated the direct implication of the allicin present in solvent-free garlic extracts obtained with ethanol (ethanolic garlic extract, EGE) and acetone (acetonic garlic extract, AGE) in the inhibition of the in-vitro growth of Helicobacter pylori (Hp), the bacterium responsible for serious gastric diseases such as ulcers and even gastric cancer. The evolution of allicin concentration as a function of time and temperature has been the subject of a kinetic study. The reaction order, activation energy, and preexponential factor (in accordance with Arrhenius theory) have been determined for the decomposition process of allicin in these organic media. First-order decomposition, an activation energy of 97.4 kJ/mol, and an Arrhenius preexponential factor of 8.9 x 10(10) s(-1) have been determined for allicin in EGE. For allicin in AGE the kinetic order determined was 1.5, the activation energy 184.5 kJ/mol, and the preexponential factor 3.1 x 10(24) s(-1) (mg/L)(-0.5). The presence or absence of allicin in these garlic products was found to be crucial for the inhibition of the in-vitro growth of Hp, as demonstrated by microbiological analysis for AGE. A relationship has been identified between the effectiveness and durability of the anti-Hp properties shown by AGE and the allicin content of these products. The bacteriostatic properties were active for up to 10 months if the samples were maintained at 6 degrees C.     

美洲大蠊粪便粗提物的聚集活性

用不同溶剂不同方法提取美洲大蠊Periplaneta americana(L.)粪便,测定美洲大蠊各虫态的诱集活性。结果表明,采用直接浸泡提取方法,丙酮、乙醇、正己烷和二氯甲烷4种粗提物对美洲大蠊各虫态都具有明显的诱集作用,其中乙醇和丙酮粗提物的引诱效果最好,正己烷次之,二氯甲烷最弱。4种溶剂粗提物对美洲大蠊雄成虫和高龄若虫聚集活性最强,对低龄若虫聚集活性最弱。用乙醇溶剂对粪便粗提,3种提取方法均对美洲大蠊有很强的诱集效果,其中索氏抽提诱集效果最弱,直接浸泡和超声波提取效果好,且差异不显著,但直接浸泡提取效果更好。     

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.     

Anti-Helicobacter pylori activity of Plumbago zeylanica L

It has been shown that the presence of infection by Helicobacter pylori is strongly associated with gastric cancer and peptic ulceration. In western medicine, a 3-fold therapeutic regimen, emphasizing the use of antibiotics, is typically used to suppress H. pylori activity. However, antibiotic drug resistance frequently develops as a consequence of such treatment. In our previous study, 50 Taiwanese folk medicinal plants were screened for their anti-H. pylori activities. The results revealed that Plumbago zeylanica L. had the highest inhibitory effects against H. pylori. In this study, therefore, we have focused on establishing the anti-H. pylori activities of P. zeylanica L. Water and the organic solvents ethanol, ethyl acetate and acetone were used for P. zeylanica L. extraction, obtaining yields of 1.66-6.84% (w/w). Excluding the water extract, higher anti-H. pylori activity was demonstrated for all the extracts, both using the agar diffusion and dilution methods. The ethyl acetate extract exhibited the lowest minimum inhibitory concentrations against five H. pylori strains, of which ranged from 0.32 to 1.28 mg ml-1, followed, in ascending order, by the acetone, ethanol and water analogs. Bactericidal activity was determined for P. zeylanica L. extracts, with the lowest minimum bactericidal concentrations (5.12-20.48 mg ml-1) demonstrated for the ethyl acetate, followed, in ascending order, by the acetone and ethanol analogs. Bactericidal activity appeared to be in a dose-dependent manner. Through a broad pH range (2-7), bactericidal activity was not affected when extract concentrations were greater than or equal to the minimum bactericidal concentration. High stability was demonstrated for the ethyl acetate P. zeylanica L. extract within pH range of 1-7, exhibiting all pH treatments bactericidal activity.     

蝉蜕活性成分的提取及其抑菌活性的研究

本文报道了对传统中药蝉蜕抑菌活性成分提取与抑菌活性研究结果。对4种提取方法和7种提取溶剂的提取效果进行了平行比较研究,研究结果显示,以体积分数95%乙醇作为提取溶剂,4种提取方法中,以超声波法提取时的浸膏得率最高。采用超声波提取法时,在7种提取溶剂中,以95%乙醇提取浸膏的得率最高。抑菌试验结果表明,不同方法获得提取物与对照组相比对Escheridia coli均有明显的抑菌作用(P≤0.05),但不同提取方法得到的提取物之间,并无显著的抑菌活性差异。其中采用超声波提取法,以95%乙醇为提取溶剂时,所得提取物的抑菌活性最高,对E.coli的最小抑菌浓度为0.078 mg/mL。该研究结果是蝉蜕抑菌成分的首次报道。     

Chemical composition and antibacterial activity of Cordia verbenacea extracts obtained by different methods

The present study describes the chemical composition and the antibacterial activity of extracts from Cordia verbenacea DC (Borraginaceae), a traditional medicinal plant that grows widely along the southeastern coast of Brazil. The extracts were obtained using different extraction techniques: high-pressure operations and low-pressure methods. The high-pressure technique was applied to obtain C. verbenacea extracts using pure CO₂ and CO₂ with co-solvent at pressures up to 30 MPa and temperatures of 30, 40 and 50 °C. Organic solvents such as n-hexane, ethyl acetate, ethanol, acetone and dichloromethane were used to obtain extracts by low-pressure processes. The antibacterial activity of the extracts was also subjected to screening against four strains of bacteria using the agar dilution method. The extraction yields were up to 5.0% w/w and up to 8.6% w/w for supercritical fluid extraction with pure CO₂ and with ethyl acetate as co-solvent, respectively, while the low-pressure extraction indicates yields up to 24.0% w/w in the soxhlet extraction using water and aqueous mixture with 50% ethanol as solvents. The inhibitory activity of the extracts in Gram-positive bacteria was significantly higher than in Gram-negative. The quantification and the identification of the extracts recovered were accomplished using GC/MS analysis. The most important components identified in the extract were artemetin, β-sitosterol, α-humulene and β-caryophyllene, among others.     

Antibacterial activity of guava,moringa, camphor bush and pelargonium extracts against bacterial wilt (Ralstonia pseudosolanacearum sp. nov.) of potato

Bacterial wilt (Ralstonia pseudosolanacearum sp. nov.) is a major disease devastating global potato production. Proposed management options are mostly expensive and ineffective. This has necessitated efforts to develop cheaper and eco-friendly management options such as use of botanicals. Antibacterial activity of ethanol and acetone plant extracts from guava (Psidium guajava), drumstick (Moringa oleifera), camphor bush (Tarchonanthus camphoratus) and pelargonium (Pelargonium zonale) against R. pseudosolanacearum sp. nov. was evaluated in-vitro at a concentration of 100 mg/mL of 1 % Dimethlysulfoxide (DMSO) using disk diffusion technique. The R. pseudosolanacearum sp. nov was isolated from infected haulms collected from potato growing field at the University of Nairobi. The most effective extracts were subjected to further screening at different concentrations to determine their minimum inhibitory concentrations (MICs). All the four plant extracts showed varied antibacterial efficacy. P. zonale leaves extract was the most effective with growth inhibition zone of 18.73 mm and 18.60 mm for ethanol and acetone solvents respectively. The average of growth inhibition zones for each plant extract was not significantly different at p ≤ 0.05 among extraction solvents. The minimum inhibitory concentration (MIC) results showed that antibacterial activity of P. zonale and P. guajava leaf started at 6.25 mg/mL with growth inhibition zones of 7.67 and 8.0 mm for ethanol and acetone solvents respectively. P. zonale and P. guajava leaf extracts exhibited significantly higher antibacterial activity at p ≤ 0.05 compared to other extracts. Thus, further research should be conducted to assess their antibacterial potency against R. pseudosolanacearum sp. nov. both in-vivo and under field condition.     

Antifungal activity of olive leaf (Olea Europaea L.) extracts from the Trilye Region of Turkey

Antimicrobial properties of olive leaf extract on some yeast were examined in this study. Fresh olive leaf extracts were prepared using various solvents (water, ethanol, acetone, ethyl acetate) in Soxhlet apparatus. Antimicrobial effects of these extacts were tested againstSaccharomyces cerevisiae ATCC 9763,Schizosaccharomyces pombe, Saccharomyces uvarum, Candida oleophila, Metschnikowia fructicola andKloeckera apiculata. The antifungal activities of these extracts were tested by the disc diffusion assay, minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC). All extracts showed various degrees of antifungal effects with 10–28 μg/ml MIC, 20–48 μg/ml MFC and 1.5–9.3 mm inhibitory zone values against yeasts utilised, except water. The results indicated that the tested yeasts were sensitive to acetone and ethyl acetate extracts. It was determined thatSaccharomyces cerevisiae ATCC 9763 was the most resistant among the yeasts.     

Microalga Scenedesmus bajacalifornicus BBKLP-07, a new source of bioactive compounds with in vitro pharmacological applications

Microalgae are photosynthetic eukaryotes which are primary producers in the food chain and also excellent sources for bioactive compounds such as alkaloids, flavonoids, phenols, saponins and other fine chemicals. In the present study, the microalga Scenedesmus bajacalifornicus BBKLP-07 was subjected to soxhlet extraction using solvents like chloroform, acetone, ethanol, methanol and aqueous solvents. All the solvents were tested for the presence of phytochemical constituents such as alkaloids, flavonoids, glycosides, phenols, lignin’s, saponins, sterols, tannins, anthraquinone and reducing sugar using the standard procedures. Furthermore, all the crude extracts were subjected to antidiabetic, antioxidant, anti-inflammatory and antimicrobial activities. Antidiabetic activity of the microalgal extracts was observed maximum in Aqueous extract. Methanolic extracts have shown maximum antioxidant activity and chloroform extracts have exhibited highest anti-inflammatory effects. Antimicrobial activities were tested against E.coli, S, typhi, C.perfringens and B.subtilis bacteria and fungi A.niger, and C. albicans. Therefore, the green microalga Scenedesmus bajacalifornicus BBKLP-07 is a rich source of biological active compounds and nutraceuticals and can be exploited for commercial applications.

     

Extraction of chlorophyll a from freshwater phytoplankton for spectrophotometric analysis

The efficiency of pigment extraction forms the crux of the spectrophotometric analysis of chlorophyll a. The alcoholic solvents, methanol and ethanol, proved to be superior to acetone and acetone with DMSO. Homogenisation and sonication did not improve the extraction in the alcoholic solvents. Boiling at 100°C had an adverse effect whereas complete extraction of the pigments was obtained at the solvents boiling point and allowing the samples to stand for 24 h in the dark.     

Continuous lactose fermentation by Clostridium acetobutylicum--assessment of acidogenesis kinetics

An assessment of the growth kinetics of acidogenic cells of Clostridium acetobutylicum DSM 792 is reported in the paper. Tests were carried out in a continuous stirred tank reactor under controlled conditions adopting a complex medium supplemented with lactose as carbon source to mimic cheese whey. The effects of acids (acetic and butyric), solvents (acetone, ethanol and butanol) and pH on the growth rate of acidogenic cells were assessed. The conversion process was characterized under steady-state conditions in terms of concentration of lactose, cells, acids, total organic carbon and pH. The growth kinetics was expressed by means of a multiple product inhibition and interacting model including a novel formulation to account for the role of pH. The model has the potential to predict microorganism growth rate under a broad interval of operating conditions, even those typical of solvents production.     

Evaluation and improvement of spectrophotometric assays of TTC reduction: maize (Zea mays) embryo as an example

The triphenyl tetrazolium chloride (TTC) reduction assay was evaluated and improved with maize seed (Zea mays cv. Zhengdan958). The reduced TTC in embryo was extracted with three kinds of organic solvents: trichloroacetic acid (TCA)/acetone, ethanol, and acetone. The absorbance spectra of the three extracts were similar, with a maximum at 485 nm. The efficiency of TCA/acetone in extracting the reduced TTC was higher than that of acetone and ethanol. A negative correlation between TTC reduction and malondialdehyde content in embryo was demonstrated. The TCA/acetone extraction may be used as a routine protocol for TTC reduction assay of seed vigor in cereal (e.g. maize, rice, wheat and barley) seeds.     

桦褐孔菌活性物质的提取工艺及体外抗糖尿病活性

对桦褐孔菌活性物质的提取工艺及其体外抗糖尿病活性进行研究。桦褐孔菌子实体用乙醇浸提后,乙醇粗提物用不同有机溶剂萃取,醇提残渣再以热水浸提,用标准曲线法测定活性组分中活性成分的含量,并检测活性物质对羟基自由基(·OH)、超氧阴离子自由基(O2-·)的清除效果以及对关键糖代谢酶α-淀粉酶、α-葡萄糖苷酶的抑制作用。结果表明:4种活性组分(乙酸乙酯相、正丁醇相、水相和粗多糖)对羟基自由基(·OH)和超氧阴离子自由基(O2-·)都有较强的清除作用,其中乙酸乙酯组分的活性最高,乙醇粗提物萃取组分对α-淀粉酶有抑制活性,而粗多糖对α-葡萄糖苷酶有抑制作用;桦褐孔菌具有抗氧化和抗糖尿病活性,其活性与活性物质种类及其含量具有相关性。     

狭叶柴胡种子萌发与内源抑制物质

分别测定了柴胡种子的水浸提液、水浸泡液与乙醚浸提液对柴胡种子萌发以及对白菜和小麦种子萌发和幼苗生长的抑制活性;用水、乙醚、乙醇和丙酮浸泡柴胡种子,比较其对种子中抑制物质的去除效果;运用原位实验方法探讨柴胡种子萌发与内源抑制物质之间的关系。结果表明：柴胡种子中含有一定活性的内源抑制物质,其对白菜幼根生长的抑制活性高于对白菜种子萌发的抑制活性,对小麦叶绿素含量、鲜质量和α-淀粉酶的活性高于对小麦种子萌发的抑制活性,并且对柴胡自身种子萌发也有明显的抑制作用;室温用乙醚浸种24 h可以提高种子发芽率。据此,建立了柴胡种子的催芽技术。     

不同实验方法检测常用有机溶剂对细菌活性的影响及其安全使用限量

【目的】采用不同实验方法测定常用有机溶剂二甲基亚砜(DMSO)、丙酮和乙醇对细菌活性的影响,以指导抗菌类药物体外抑菌实验所用溶剂的选择和添加限量。【方法】采用常规体外抑菌实验方法(纸片扩散法、肉汤稀释法),并参照生长曲线法检测有机溶剂DMSO、丙酮和乙醇对大肠杆菌(Escherichia coli)及金黄色葡萄球菌(Staphylococcus aureus)的抑菌作用,采用扫描电子显微镜(SEM)观察溶剂作用后的细菌形态变化。【结果】3种溶剂对E.coli和S.aureus抑菌率达到20%时,在肉汤稀释法下,DMSO、丙酮、乙醇的浓度(体积比)分别为1.00%、0.25%、2.00%和1.00%、1.00%、0.50%;在生长曲线法下,溶剂浓度(体积比)分别为0.50%、1.00%、0.50%和1.00%、0.50%、0.50%;而在纸片扩散法下,32%(体积比)DMSO和32%(体积比)乙醇对E.coli产生明显抑菌圈,但3种溶剂对S.aureus均无抑菌圈出现。3种方法比较后得出:当3种溶剂的抑菌率达到20%时,溶剂浓度(体积比)均低于0.5%,对细菌整体生长活性影响较小。SEM结果表明控制溶剂使用限量可有效减少其对E.coli生长过程的影响。【结论】相对于DMSO和丙酮,乙醇对微生物生长繁殖能力的影响更加明显;采用相同浓度有机溶剂时,液态条件下(肉汤稀释法和生长曲线法)微生物受到有机溶剂的影响较大。     

A comparison between two methods of extracting chlorophyll-a from different phytoplankton samples

Summary A comparison has been made between chlorophyll-a estimations based on the cold acetone extraction procedure and on the hot ethanol extraction method. When comparing the results obtained with both methods a standard deviation of 20% must be taken into account. A correlation between extraction efficiency and phytoplankton species composition could not be found. Although the chlorophyll-a yield in ethanol extracts is not better than that in acetone extracts, there are several advantages in using the ethanol method, particularly the short extraction period and the cheaper procedure.     

蛹虫草无性型菌丝体提取液体外抗氧化活性研究

分别测定了蛹虫草无性型菌丝体不同溶剂提取液在不同抗氧化模型中的抗氧化作用。结果表明,各种提取液对二苯基苦味酰基苯肼自由基(·DPPH)和羟自由基(·OH)均有显著的清除作用:在50mg/mL时,去离子水、70%乙醇和70%丙酮提取物对DPPH自由基的清除率分别为89.2%、83.6%和75.9%;70%乙醇提取物在30mg/mL时对·OH自由基的清除率达到100%;在50mg/mL时70%丙酮和去离子水提取物的·OH自由基清除率分别为95.6%和89.6%。在一定浓度下,各提取液对邻苯三酚自氧化也均有抑制作用;不同溶剂提取物的还原能力强弱为:70%乙醇>去离子水>70%丙酮。各种提取物的抗氧化组分不同,提取物浓度与抗氧化性成正相关。     

Inhibitory effect of Torilis anthriscus on growth of microorganisms

Antibacterial and antifungal activities of aqueous, ethanol and ethyl acetate extract of Torilis anthriscus (L.) Gmel. (Apiaceae) were tested in vitro against ten species of bacteria and five of fungi. Antimicrobial properties were determined by disk diffusion and broth tube dilution method. In the minimum inhibitory concentrations (MICs), the ethanol extract showed the highest activity, followed by the ethyl acetate extract and the aqueous extract against bacterial species, while the extracts were inactive against the tested fungi species. The most active extract was chosen to examine the effects of its combinations with commercial antibiotics by checkerboard method. The obtained results showed that the interactions between ethanol extract/streptomycin and ethanol extract/chloramphenicol were additive and indifferent against the tested human-pathogenic bacteria. Synergism and antagonism were not observed.     

Efficacy of cashew kernel extracts in the control of the maize weevil,Sitophilus zeamais (Coleoptera: Curculionidae)

Six different solvents namely: methanol, ethanol, water, acetone, pet-ether or n-hexane was used to extract the essential oils from cashew kernel. The extracted oils were evaluated on Sitophilus zeamais (Motschulsky 1855) for mortality, oviposition and adult emergence effects. The long-term storage and water absorption capacity of the treated maize grains was also investigated. Results showed that the steam distillate was most lethal on S. zeamais. The least potent extracts on S. zeamais were those extracted with methanol and ethanol. The extracts drastically reduced oviposition in S. zeamais. Maize grains treated with steam distillate extract had the lowest water absorption capacity, while the highest water absorption capacity was obtained in grains treated with ethanol and methanol. Extracts of steam distillate, n-hexane, petroleum ether and acetone completely prevented infestation and subsequence damage of the treated maize grains for a period of three months. In the grains treated with the methanol and ethanol extracts, damage of 28.61% and 6.24%, respectively were obtained. The results obtained reveal that extracts of cashew kernel are effective in controlling S. zeamais and could serve as an alternative to synthetic insecticides.