Screening of the Dichloromethane: Methanolic Extract of Centella asiatica for Antibacterial Activities against Salmonella typhi, Escherichia coli, Shigella sonnei, Bacillus subtilis, and Staphylococcus aureus.

Bacterial infections are responsible for a large number of deaths every year worldwide. On average, 80% of the African population cannot afford conventional drugs. Moreover, many synthetic antibiotics are associated with side effects and progressive increase in antimicrobial resistance. Currently, there is growing interest in discovering new antibacterial agents from ethnomedicinal plants. About 60% of the population living in developing countries depends on herbal drugs for healthcare needs. This study involved the screening of Centella asiatica commonly used by herbal medicine practitioners in Kisii County to treat symptoms related to bacterial infections. Standard bioassay methods were applied throughout the study. They included preliminary screening of dichloromethane: methanolic extract of Centella asiatica against human pathogenic bacteria including Salmonella typhi ATCC 19430, Escherichia coli ATCC 25922, Shigella sonnei ATCC 25931, Bacillus subtilis ATCC 21332, and Staphylococcus aureus ATCC 25923 using agar disc diffusion, broth microdilution method, and time-kill kinetics with tetracycline as a positive control. Phytochemical screening was carried out to determine the different classes of compounds in the crude extracts. Data were analyzed using one way ANOVA and means separated by Tukey's test. Dichloromethane: methanolic extract of Centella asiatica was screened against the selected bacterial strains. Time-kill kinetic studies of the extracts showed dose- and time-dependent kinetics of antibacterial properties. Phytochemical screening of the DCM-MeOH extract revealed the presence of alkaloids, flavonoids, phenolics, terpenoids, cardiac glycosides, saponins, steroids, and tannins. The present study indicates that the tested plant can be an important source of antibacterial agents and recommends that the active phytoconstituents be isolated, identified, and screened individually for activities and also subjected further for in vivo and toxicological studies.

Satureja hortensis induces cell death and inhibited cell cycle progression in K562 myelogenous and Jurkat T cell leukemia cell lines.

Several plants of Satureja genus have shown anti-tumor activity. We investigated the antileukemia effects of different fractions of Satureja hortensis (Summer savory). The growth inhibitory effect of S. hortensis fractions on K562 and Jurkat leukemia cells were determined by MTT assay. The most effective fractions were analyzed by flow cytometry and colorimetric assay for apoptosis induction and cell cycle changes. Various fractions from S. hortensis showed growth inhibitory effects on leukemia cells, among them two hexane and dichloromethane fractions with IC50 values of 32.1-47.8 µg/ml (K562) and 44.3-45.7 µg/ml (Jurkat) were the most effective. According to annexin V staining, both of these fractions significantly induced apoptosis at 50µg/ml in K562 (hexane; 73.06 ± 5.11% and dichloromethane; 96.14 ± 2.33%) and Jurkat cells (hexane; 78.85 ± 11.9% and dichloromethane; 94.05 ± 2.47%) 48 h after treatment. They increased cell accumulation in sub-G1 phase (>50%, p < .001) and decreased number of cells in G0-G1, S and G2M phases. The fractions significantly increased the caspase-3 activity in both cell lines (≈2.5-3.5 fold of untreated cells). Hexane and dichloromethane fractions of S. hortensis had the capacity to induce death and change the cell cycle distribution in leukemia cells; therefore they might be good candidates for more studies in regard to their possible therapeutic usefulness in leukemia.

Coated mesh photocatalytic reactor for air treatment applications: comparative study of support materials.

An experimental comparative study of different meshes as support materials for photocatalytic applications in gas phase is presented. The photocatalytic oxidation of dichloromethane in air was addressed employing different coated meshes in a laboratory-scale, continuous reactor. Two fiberglass meshes and a stainless steel mesh were studied regarding the catalyst load, adherence, and catalytic activity. Titanium dioxide photocatalyst was immobilized on the meshes by dip-coating cycles. Results indicate the feasibility of the dichloromethane elimination in the three cases. When the number of coating cycles was doubled, the achieved conversion levels were increased twofold for stainless steel and threefold for the fiberglass meshes. One of the fiberglass meshes (FG2) showed the highest reactivity per mass of catalyst and per catalytic surface area.

Modified rotating biological contactor for removal of dichloromethane vapours.

Bioreactors are used for the treatment of waste gas and odour that has gained much acceptance in the recent years to treat volatile organic compounds (VOCs). The different types of bioreactors (biofilter, biotrickling filter and bioscrubber) have been used for waste gas treatment. Each of these reactors has some advantages and some limitations. Though biodegradation is the main process for the removal of the pollutants, the mechanisms of removal and the microbial communities may differ among these bioreactors. Consequently, their performance or removal efficiency may also be different. Clogging of reactor and pressure drop are the main problems. In this study attempts are made to use the principle of rotating biological contactor (RBC) used for wastewater treatment for the removal of VOC. To overcome the above problem the RBC is modified which is suitable for the treatment of VOC (dichloromethane, DCM). DCM is harmful to human health and hazardous to the atmospheric environment. Modified RBC had no clogging problems and no pressure drop. So, it can handle the pollutant load for a longer period of time. A maximum elimination capacity of 25.7 g/m3 h has been achieved in this study for the DCM inlet load of 58 g/m3 h. The average biofilm thickness is 1 mm. The transient behaviour of the modified RBC treating DCM was investigated. The modified RBC is able to handle shutdown, restart and shock loading operations.

Xanthone-rich dichloromethane fraction of Securidaca inappendiculata, the possible antirheumatic material base with anti-inflammatory, analgesic, and immunodepressive effects.

CONTEXT: Securidaca inappendiculata Hassk. is an traditional Chinese medicine curing rheumatoid arthritis, but there is a lack of reports on material base research. OBJECTIVE: To find the active fraction of S. inappendiculata contributing the most to antirheumatic activity. MATERIALS AND METHODS: Prior to assays in vivo, mice were treated with different fractions from S. inappendiculata for 5 d at doses relative to 10, 5, and 2.5 g/kg of crude drug. Hot plate test and carrageenan-induced paw edema test were used to investigate analgesic and anti-inflammatory activities. PGE2 levels in inflammatory paws were determined by a colorimetric method. Carbon clearance test in vivo and lymphocyte transformation test in vitro were employed to assess the immune regulation activity. HPLC was used to explore the main compounds in the active fraction. RESULTS: All the fractions, especially the dichloromethane fraction (SID), alleviated inflammation. High dose of SID (112 mg/kg) inhibited paw swelling by 63.1%, and decreased PGE2 level to 38 ng/mL. The ethyl acetate fraction (SIE) and SID suppressed the carbon clearance rate (K = 0.044, 0.038 for high dose) efficiently. All fractions hindered the transformation and proliferation of lymphocyte, and prolonged the reaction time of rats in the hot plate test. The concentrations of two typical xanthones: 2-hydroxyl-1,7-dimethoxyl-xanthone and 1,7-dihydroxyl-xanthone in SID were 0.93% and 1.19%, respectively, by HPLC analysis. CONCLUSION: SID exhibited significant anti-inflammatory, analgesic, and immunodepressive effects in vivo and vitro, and deemed as the main material base for the antirheumatic activity.

Anti-Mycobacterium tuberculosis activity and cytotoxicity of Calophyllum brasiliense Cambess (Clusiaceae)

We evaluated the in vitro anti-Mycobacterium tuberculosis activity and the cytotoxicity of dichloromethane extract and pure compounds from the leaves of Calophyllum brasiliense. Purification of the dichloromethane extract yielded the pure compounds (-) mammea A/BB (1), (-) mammea B/BB (2) and amentoflavone (3). The compound structures were elucidated on the basis of spectroscopic and spectrometric data. The contents of bioactive compounds in the extracts were quantified using high performance liquid chromatography coupled to an ultraviolet detector. The anti-M. tuberculosis activity of the extracts and the pure compounds was evaluated using a resazurin microtitre assay plate. The cytotoxicity assay was performed in J774G.8 macrophages using the 3-(4,5-dimethyl thiazol-2-yl)-2,5-diphenyl tetrazolium bromide colourimetric method. The quantification of the dichloromethane extract showed (1) and (2) at concentrations of 31.86 ± 2.6 and 8.24 ± 1.1 µg/mg of extract, respectively. The dichloromethane and aqueous extracts showed anti-M. tuberculosis H37Rv activity of 62.5 and 125 µg/mL, respectively. Coumarins (1) and (2) showed minimal inhibitory concentration ranges of 31.2 and 62.5 µg/mL against M. tuberculosis H37Rv and clinical isolates. Compound (3) showed no activity against M. tuberculosis H37Rv. The selectivity index ranged from 0.59-1.06. We report the activity of the extracts and coumarins from the leaves of C. brasiliense against M. tuberculosis.

Therapeutic effects of dichloromethane fraction of Securidaca inappendiculata on adjuvant-induced arthritis in rat.

ETHNOPHARMACOLOGICAL RELEVANCE: Securidaca inappendiculata (SI) is a traditional antirheumatic medicine used in China. The present study was designed to investigate the therapeutic efficacy of dichloromethane fraction of SI (SID) at three different doses on adjuvant induced arthritis (AA) rats. METHODS: Arthritis severity was evaluated by arthritic score, body weight loss, paw circumference, histological changes and hyperplasia of lymphatic tissues. Serum samples were collected for estimation of superoxide dismutase (SOD), glutathione (GSH), hydroxy radical (OH·), nitric oxide (NO), malondialdehyde (MDA), N-acetyl glucosaminidase (NAG), sialic acid (SA), alkaline phosphatase (ALP), aspartate transaminase (AST) and alanine transaminase (ALT). The levels of GSH, MDA, NAG and SA in liver were also assessed. The levels of interleukin-1 (IL-1), tumor necrosis factor alpha (TNF-α), monocyte chemotactic protein 1 (MCP-1) and vascular endothelial growth factor (VEGF) were determined using ELISA method. Another portion of blood was used for total and differential leucocyte counts. RESULTS: Administration with SID (at high dose with 100 mg/kg) significantly ameliorated the AA severity, suggested by the modulatory effects on body weight loss, paw swelling, hyperplasia of lymphatic tissues and synovial membrane, neutrocytosis and lymphocytosis. It also decreased levels of NO, MDA and OH·, restored SOD and GSH levels in serum. The abnormal increased levels of AST, ALT, ALP, NAG and SA significantly were reverted (compared with AA rats, P<0.01). A similar result was observed in livers. Levels of IL-1, TNF-α, MCP-1 and VEGF were reduced dramatically by SID too. CONCLUSION: The results suggest SID possesses substantial anti-arthritic activity. The therapeutic efficacy may be due to immumodepressive effects, cytokines regulation, increasing membrane stability and antioxidantive activity.

Anti-Mycobacterium tuberculosis activity and cytotoxicity of Calophyllum brasiliense Cambess (Clusiaceae).

We evaluated the in vitro anti-Mycobacterium tuberculosis activity and the cytotoxicity of dichloromethane extract and pure compounds from the leaves of Calophyllum brasiliense. Purification of the dichloromethane extract yielded the pure compounds (-) mammea A/BB (1), (-) mammea B/BB (2) and amentoflavone (3). The compound structures were elucidated on the basis of spectroscopic and spectrometric data. The contents of bioactive compounds in the extracts were quantified using high performance liquid chromatography coupled to an ultraviolet detector. The anti-M. tuberculosis activity of the extracts and the pure compounds was evaluated using a resazurin microtitre assay plate. The cytotoxicity assay was performed in J774G.8 macrophages using the 3-(4,5-dimethyl thiazol-2-yl)-2,5-diphenyl tetrazolium bromide colourimetric method. The quantification of the dichloromethane extract showed (1) and (2) at concentrations of 31.86 ± 2.6 and 8.24 ± 1.1 µg/mg of extract, respectively. The dichloromethane and aqueous extracts showed anti-M. tuberculosis H37Rv activity of 62.5 and 125 µg/mL, respectively. Coumarins (1) and (2) showed minimal inhibitory concentration ranges of 31.2 and 62.5 µg/mL against M. tuberculosis H37Rv and clinical isolates. Compound (3) showed no activity against M. tuberculosis H37Rv. The selectivity index ranged from 0.59-1.06. We report the activity of the extracts and coumarins from the leaves of C. brasiliense against M. tuberculosis.

Dichloromethane removal and microbial variations in a combination of UV pretreatment and biotrickling filtration.

Biofiltration of hydrophobic and/or recalcitrant volatile organic compounds in industry is currently limited. A laboratory-scale system integrating ultraviolet (UV) photodegradation and a biotrickling filter (BTF) was developed to treat dichloromethane (DCM), and this was compared to BTF alone. A combined UV-BTF approach permitted faster biofilm formation and greater removal than BTF. DCM distribution and its photodegradation intermediates revealed that the lower filter of the UV-BTF contributed more to CO2 production; the upper filter assisted more with DCM removal. The UV-BTF kept secretion of extracellular polymeric substances at a normal level with an evenly distributed biomass. Pyrosequencing analysis showed that the dominant population in the combined biofilter was more diverse than that in BTF alone. Our data provide a foundation for understanding the effect of UV pretreatment on BTF performance and the microbial community.

Remediation of dichloromethane (CH2Cl2) using non-thermal, atmospheric pressure plasma generated in a packed-bed reactor.

This work describes the application of a non-thermal plasma generated in a dielectric barrier packed-bed plasma reactor for the remediation of dichloromethane (CH2Cl2, DCM). The overall aim of this investigation is to identify the role of key process parameters and chemical mechanisms on the removal efficiency of DCM in plasma. The influence of process parameters, such as oxygen concentration, concentration of initial volatile organic compounds (VOCs), energy density, plasma residence time, and background gas, on the removal efficiency of 500 ppm DCM was investigated. Results showed a maximum removal efficiency with the addition of 2-4% oxygen into a nitrogen plasma. It is thought that oxygen concentrations in excess of 4% decreased the decomposition of chlorinated VOCs as a result of ozone and nitrogen oxide formation. Increasing the residence time and the energy density resulted in increasing the removal efficiency of chlorinated VOCs in plasma. A chemical kinetic model has been developed on the basis of the proposed reaction scheme, and the calculation of end product concentrations are in general good agreement with the observed values. With the understanding of the effect of the key parameters, it has been possible to optimize the remediation process.

Comparison of agitake (Pleurotus eryngii var. ferulae) volatile components with characteristic odors extracted by hydrodistillation and solvent-assisted flavor evaporation.

The chemical composition of volatile oil from agitake (Pleurotus eryngii var. ferulae) was established for the first time using gas chromatography (GC) and GC-mass spectrometry. Sixty-seven and 24 components were extracted by hydrodistillation (HD) using diethyl ether (DE) and dichloromethane (DM), respectively; these components accounted for 80.3% and 91.8% of the total oil, respectively. Thirteen and 48 components of were extracted by the solvent-assisted flavor evaporation method (SAFE), using DE and DM, respectively, and identified; these components accounted for 83.5% and 82.0% of the total oil, respectively. Methylsuccinimide and 2,3,7-trimethyl-2-octene were the most characteristic components by SAFE using DM.Odor evaluation of the volatile oil from agitake was also carried out using GC-olfactometry (GC-O), aroma extraction dilution analysis (AEDA), and the odor activity value (OAV). Sixteen, 8, 5 and 9 aroma-active components were identified using HD (DE and DM) and SAFE (DE and DM), respectively. The main aroma-active components extracted using HD and SAFE were 1-octen-3-ol (mushroom-like) and phenylacetaldehyde (floral), respectively. This study proved that HD and SAFE can be used as complementary extraction techniques for the complete characterization of volatile oil from agitake.

[Removal of mixed waste gases by the biotrickling filter].

A biotrickling filter (BTF) was designed for treating mixed waste gases, which contained hydrogen sulfide (H2S), tetrahydrofuran (THF) and dichloromethane (DCM) at the start-up and steady states. The removal efficiency of H2S and DCM could maintain about 99% and 60%, respectively, and the removal efficiency of DCM was reduced from 90% to 37% with the shortening empty bed retention time (EBRT) form 50 to 20 seconds when the inlet concentrations were 200, 100, 100 mg x m(-3) of H2S, THF, DCM, respectively. In the theoretical study, the biodegradation efficiency of contaminants was H2S > THF > DCM by analyzing the Michaelis-Menten Dynamic model.

Evaluation of larvicidal activity and brine shrimp toxicity of rhizome extracts of Zingiber zerumbet (L.) Smith.

INTRODUCTION: In this study, we used dichloromethane (DCM) and methanol (MeOH) extracts of the Zingiber zerumbet rhizome to evaluate brine shrimp lethality and larvicidal activity on Aedes aegypti and Anopheles nuneztovari mosquitoes. METHODS: Bioassays were performed by exposing third-instar larvae of each mosquito species to the DCM or MeOH extracts. RESULTS: Probit analysis with DCM and MeOH extracts demonstrated efficient larvicidal activity against A. aegypti and A. nuneztovari larvae. CONCLUSIONS: The DCM and MeOH extracts showed higher activity against A. nuneztovari larvae than against A. aegypti larvae, suggesting that the extracts have species-specific activity.

Evaluation of larvicidal activity and brine shrimp toxicity of rhizome extracts of Zingiber zerumbet (L.) Smith

Introduction In this study, we used dichloromethane (DCM) and methanol (MeOH) extracts of the Zingiber zerumbet rhizome to evaluate brine shrimp lethality and larvicidal activity on Aedes aegypti and Anopheles nuneztovari mosquitoes. Methods Bioassays were performed by exposing third-instar larvae of each mosquito species to the DCM or MeOH extracts. Results Probit analysis with DCM and MeOH extracts demonstrated efficient larvicidal activity against A. aegypti and A. nuneztovari larvae. Conclusions The DCM and MeOH extracts showed higher activity against A. nuneztovari larvae than against A. aegypti larvae, suggesting that the extracts have species-specific activity. .

Evaluation of antimicrobial activity of extracts of Tibouchina candolleana (melastomataceae), isolated compounds and semi-synthetic derivatives against endodontic bacteria

This work describes the phytochemical study of the extracts from aerial parts of Tibouchina candolleana as well as the evaluation of the antimicrobial activity of extracts, isolated compounds, and semi-synthetic derivatives of ursolic acid against endodontic bacteria. HRGC analysis of the n-hexane extract of T. candolleana allowed identification of b-amyrin, a-amyrin, and b-sitosterol as major constituents. The triterpenes ursolic acid and oleanolic acid were isolated from the methylene chloride extract and identified. In addition, the flavonoids luteolin and genistein were isolated from the ethanol extract and identified. The antimicrobial activity was investigated via determination of the minimum inhibitory concentration (MIC) using the broth microdilution method. Amongst the isolated compounds, ursolic acid was the most effective against the selected endodontic bacteria. As for the semi-synthetic ursolic acid derivatives, only the methyl ester derivative potentiated the activity against Bacteroides fragilis.

Hazardous dichloromethane recovery in combined temperature and vacuum pressure swing adsorption process.

Organic vapors emitted from solvents used in chemical and pharmaceutical processes, or from hydrocarbon fuel storage stations at oil terminals, can be efficiently captured by adsorption onto activated carbon beds. To recover vapors after the adsorption step, two modes of regeneration were selected and could be possibly combined: thermal desorption by hot nitrogen flow and vacuum depressurization (VTSA). Because of ignition risks, the conditions in which the beds operate during the adsorption and regeneration steps need to be strictly controlled, as well as optimized to maintain good performances. In this work, the optimal conditions to be applied during the desorption step were determined from factorial experimental design (FED), and validated from the process simulation results. The regeneration performances were compared in terms of bed regeneration rate, concentration of recovered volatile organic compounds (VOC) and operating costs. As an example, this methodology was applied in case of dichloromethane. It has been shown that the combination of thermal and vacuum regeneration allows reaching 82% recovery of dichloromethane. Moreover, the vacuum desorption ended up in cooling the activated carbon bed from 93°C to 63°C and so that it significantly reduces the cooling time before starting a new cycle.

Antimicrobial activity of the dichloromethane extract from in vitro cultured roots of Morinda royoc and its main constituents.

The dichloromethane extract and seven anthraquinones isolated from in vitro cultured roots of Morinda royoc L. were tested for their antimicrobial activity against seven yeast and seven bacterial strains. The extract showed a minimum inhibitory concentration (MIC) value of 15.6 microg/m against all species of Candida tested; except C. glabrata (MIC 1.95 microg/mL), and it inhibited the growth of oxacillin-resistant Staphylococcus aureus (MIC 31.2 microg/mL). Only morindone showed activity against all yeast strains (MIC 1.9 microg/mL), and against oxacillin-resistant Staphylococcus aureus (MIC 15 microg/mL).

Distillation in microchemical systems using capillary forces and segmented flow.

Distillation is a ubiquitous method of separating liquid mixtures based on differences in volatility. Performing such separations in microfluidic systems is difficult because interfacial forces dominate over gravitational forces. We describe distillation in microchemical systems and present an integrated silicon device capable of separating liquid mixtures based on boiling point differences. Microfluidic distillation is realized by establishing vapor-liquid equilibrium during segmented flow. Enriched vapor in equilibrium with liquid is then separated using capillary forces, and thus enabling a single-stage distillation operation. Design criteria for operation of on-chip distillation is set forth, and the working principle demonstrated by separation of binary mixtures of 50 : 50 mol% MeOH-toluene and 50 : 50 mol% DCM-toluene at 70.0 degrees C. Analysis of vapor condensate and liquid exiting a single-stage device gave MeOH mole fractions of 0.22 +/- 0.03 (liquid) and 0.79 +/- 0.06 (vapor). Similarly, DCM mole fractions were estimated to be 0.16 +/- 0.07 (liquid) and 0.63 +/- 0.05 (vapor). These experimental results were consistent with phase equilibrium predictions.

Removal of dichloromethane from waste gases in one- and two-liquid-phase stirred tank bioreactors and biotrickling filters.

The removal of dichloromethane (DCM) from polluted air was studied both in biotrickling filters and in continuous stirred tank bioreactors, using either a single-liquid aqueous phase or a combination of an aqueous-organic liquid phase. The presence of the organic phase, i.e. silicone oil, at a volume ratio of 10% of the liquid phase, increased the maximum EC by about 25% in the BTF, reaching 200 gm(3)/h, and by as much as 300% in the CSTB, reaching 350 gm(3)/h. Based on data of chloride release in the aqueous phase and carbon dioxide production in the gas phase, complete dechlorination and mineralization of the pollutant could be confirmed. When applying shock loads, a more stable behaviour was observed in the presence of the organic phase. Generally, the completely mixed reactors were also more stable than the plug-flow biotrickling filters, irrespective of the presence of the organic phase. The use of molecular techniques allowed showing that the originally inoculated DCM-degrading Hyphomicrobium strains remained present, although not dominant, after long-term bioreactor operation. Different new bacterial populations did also appear in the systems, some of which were unable to degrade DCM.

Preparation of a new chiral stationary phase for HPLC based on the (R)- 1-phenyl-2-(4-methylphenyl)ethylamine amide derivative of (S)-valine and 2-chloro-3,5-dinitrobenzoic acid: enantioseparation of amino acid derivatives and pyrethroid insecticides.

A novel chiral stationary phase (CSP) for HPLC was prepared by bonding (R)-1-phenyl-2-(4-methylphenyl)ethylamine amide derivative of (S)-valine to aminopropyl silica gel through a 2-amino-3,5-dinitro-1-carboxamido-benzene unit. The CSP was used for the separation of some amino acid derivatives and pyrethroid insecticides by chiral HPLC. Satisfactory baseline separation required optimization of the variables of mobile phase composition. Use of dichloromethane as modifier in the mobile phase gave baseline separations of amino acid derivatives. The two enantiomers of fenpropathrin and four stereoisomers of fenvalerate were baseline separated using hexane-dichloromethane-ethanol as mobile phase. The results show that the enantioselectivity of the new CSP is better than Pirkle type 1-A column for these compounds. Only partial separations were observed for the stereoisomers of cypermethrin and cyfluthrin, which gave even and eight peaks, respectively.