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PubMed2002

Host selection as a downstream strategy: polyelectrolyte precipitation of beta-glucuronidase from plant extracts.

Host selection can be a strategy to simplify downstream processing for protein recovery. Advancing capabilities for using plants as hosts offers new host opportunities that have received only limited attention from a downstream processing perspective. Here, we investigated the potential of using a polycationic precipitating agent (polyethylenimine; PEI) to precipitate an acidic model protein (beta-glucuronidase; GUS) from aqueous plant extracts. To assess the potential of host selection to enhance the ease of recovery, the same procedure was applied to oilseed extracts of canola, corn (germ), and soy. For comparison, PEI precipitation of GUS was also evaluated from a crude bacterial fermentation broth. Two versions of the target protein were investigated--the wild-type enzyme (WTGUS) and a genetically engineered version containing 10 additional aspartates on each of the enzyme's four homologous subunits (GUSD10). It was found that canola was the most compatible expression host for use with this purification technique. GUS was completely precipitated from canola with the lowest dosage of PEI (30 mg PEI/g total protein), and over 80% of the initial WTGUS activity was recovered with 18-fold purification. Precipitation from soy gave yields over 90% for WTGUS but only 1.3-fold enrichment. Corn, although requiring the most PEI relative to total protein to precipitate (210 mg PEI/g total protein for 100% precipitation), gave intermediate results, with 81% recovery of WTGUS activity and a purification factor of 2.6. The addition of aspartate residues to the target protein did not enhance the selectivity of PEI precipitation in any of the systems tested. In fact, the additional charge reduced the ability to recover GUSD10 from the precipitate, resulting in lower yields and enrichment ratios compared to WTGUS. Compared to the bacterial host, plant systems provided lower polymer dosage requirements, higher yields of recoverable activity and greater purification factors.

Chemical Precipitation↗

PubMed1987

Acid-labile sulfide and zero-valence sulfur in plant extracts containing chlorophyll and ionic detergents.

Two methods for analysis of acid-labile sulfide and zero-valence sulfur in plant extracts containing chlorophyll as well as ionic and/or nonionic detergents are presented. Both methods are based on the conversion of sulfide into methylene blue. In the first method an ethyl acetate extraction step is used to remove chlorophyll and its degradation products which otherwise prevent spectrophotometric quantitation of methylene blue. The second assay method employs 35S-labeled plant extracts. This method, which involves thin-layer chromatography and autoradiography, is potentially more sensitive than the spectrophotometric assay in detecting acid-labile sulfide and zero-valence sulfur.

Acetates↗

PubMed1998

An in vivo model to study the anti-malaric capacity of plant extracts.

An in vivo model to study the antimalaric effect of plant extracts is described. White mice (25-30 g body weight) are treated subcutaneously with 0.6 ml of the diluted extract starting seven days before P. berghei infection; treatment continues until death or for 30 days. Simultaneously 0.2 ml of the extract are applied per os starting three days before infection. In a test of the model, treated and non-treated animals differed in body weight, survival time, haematocrite, parasitemia development, and spleen or liver weight of recent dead or killed mice.

Animals↗

PubMed2006

Screening of antihelminthic effects of Indian plant extracts: a preliminary report.

INTRODUCTION: The purpose of the present study was to evaluate the antihelminthic effect of plants from the Ayurvedic system of medicine traditionally used in India. MATERIALS AND METHODS: Six plant extracts were assayed for their activity against free-living nematodes. Inhibitory effects on free-living nematodes were evaluated in vitro using aqueous or ethanolic extracts. CONCLUSIONS: Of six plants assayed, Momordica charantia yielded the best results, its crude extract producing 96% mortality.

Animals↗

PubMed2006

Antibacterial activity of plant extracts from Brazilian southeast region.

A screening was conducted with 26 plants collected in the Brazilian southeast region, to identify plant extracts with antibacterial properties against Aeromonas hydrophila, Bacillus subtilis, Pseudomonas aeruginosa and Staphylococcus aureus. Initially, the agar diffusion method was employed. Then, those extracts presenting activity were submitted to a broth microdilution assay to determine the minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC). It was observed that 13 of the tested extracts showed antibacterial activity. The best results were obtained with those from Lantana lilacina and Phyllanthus tenellus.

Anti-Bacterial Agents↗

PubMed2004

HPLC-based bioactivity profiling of plant extracts: a kinetic assay for the identification of monoamine oxidase-A inhibitors using human recombinant monoamine oxidase-A.

An assay for the HPLC-based search for monoamine oxidase-A (MAO-A) inhibitors in plant extracts was established. It combines human recombinant MAO-A, expressed as GST-fusion protein in yeast, with a kinetic measurement of the conversion of kynuramine to 4-hydroxyquinoline. Substrate selectivity and kinetic parameters of the GST-fusion protein were comparable to the wild-type enzyme. The applicability of the assay to HPLC-based activity profiling was tested with plant extracts spiked with small amounts of known MAO inhibitors.

Chromatography, High Pressure Liquid↗

PubMed2002

Intact glucosinolate analysis in plant extracts by programmed cone voltage electrospray LC/MS: performance and comparison with LC/MS/MS methods.

We present a comprehensive, sensitive, and highly specific negative ion electrospray LC/MS method for identifying all structural classes of glucosinolates in crude plant extracts. The technique is based on the observation of simultaneous maxima in the abundances of the m/z 96 and 97 ions, generated by programmed cone voltage fragmentation, in the mass chromatogram. The abundance ratios lie in the range 1:2-1:4 ([m/z 96]/[m/z 97]). Examination of the corresponding full-scan mass spectra allows individual glucosinolates of all structural classes to be identified rapidly and with confidence. The use of linearly programmed cone voltage fragmentation enhances characteristic fragment ions without compromising the abundance of the analytically important [M - H]- ion and its associated (and analytically useful) sulfur isotope peaks. Detection limits are in the low nanogram range for full-scan, programmed cone voltage spectra. Comparison of the technique with LC/MS/MS methods (product ion, precursor ion, and constant neutral loss scans) has shown that the sensitivity and selectivity of the programmed cone voltage method is superior. Data obtained on a variety of plant extracts confirmed that the methodology was robust and reliable.

Chromatography, High Pressure Liquid↗

PubMed2004

Effects of natural plant extracts on ruminal protein degradation and fermentation profiles in continuous culture.

Eight dual-flow continuous culture fermenters were used in four consecutive periods of 10 d to study the effects of six natural plant extracts on ruminal protein degradation and fermentation profiles. Fermenters were fed a diet with a 52:48 forage:concentrate ratio (DM basis). Treatments were no extract (CTR), 15 mg/kg DM of a mixture of equal proportions of all extracts (MIX), and 7.5 mg/kg DM of extracts of garlic (GAR), cinnamon (CIN), yucca (YUC), anise (ANI), oregano (ORE), or pepper (PEP). During the adaptation period (d 1 through 8), samples for ammonia N and VFA concentrations were taken 2 h after feeding. On d 9 and 10, samples for VFA (2 h after feeding), and peptide, AA, and ammonia N concentrations (0, 2, 4, 6, and 8 h after feeding) were also taken. Differences were declared at P < 0.05. During the adaptation period, total VFA and ammonia N concentrations were not affected by treatments. The acetate proportion was higher from d 2 to 6 in CIN, GAR, ANI, and ORE, and the propionate proportion was lower from d 2 to 4 in CIN and GAR, and from d 2 to 5 in ANI and ORE, compared with CTR. However, the proportion of individual VFA (mol/100 mol) was similar in all treatments after d 6, except for valerate in d 9 and 10, which was lower in PEP (2.8 +/- 0.27) compared with CTR (3.5 +/- 0.27). The average peptide N concentration was 31% higher in MIX, and 26% higher in CIN and YUC compared with CTR (6.5 +/- 1.07 mg/100 mL). The average AA N concentration was 17 and 15% higher in GAR and ANI, respectively, compared with CTR (7.2 +/- 0.77 mg/100 mL). The average ammonia N concentration was 31% higher in ANI and 25.5% lower in GAR compared with CTR (5.5 +/- 0.51 mg/100 mL). The accumulation of AA and ammonia N in ANI suggested that peptidolysis and deamination were stimulated. The accumulation of AA N and the decrease in ammonia N in GAR suggests that deamination was inhibited. The accumulation of peptide N and the numerical decrease in AA N in CIN suggest that peptidolysis was inhibited. Results indicate that plant extracts modified ruminal fermentation, but microbes were adapted to some extracts after 6 d of fermentation. Therefore, data from short-term in vitro fermentation studies may lead to erroneous conclusions, and should be interpreted with caution. Careful selection of these additives may allow the manipulation of protein degradation in the rumen.

Acetates↗

PubMed2006

Management of canine atopic dermatitis using the plant extract PYM00217: a randomized, double-blind, placebo-controlled clinical study.

This study evaluated PYM00217, a proprietary blend of plant extracts, in the management of canine atopic dermatitis (AD). One hundred and twenty dogs were diagnosed with perennial AD on the basis of history, clinical signs, a positive test for perennial allergens and elimination of other dermatoses. Exclusion criteria included antimicrobials within 7 days, antihistamines within 14 days, oral/topical glucocorticoids or ciclosporin within 28 days, and parenteral glucocorticoids, essential fatty acids or immunotherapy within 56 days. Flea control, shampoos and ear cleaners were permitted. Dogs with a minimum canine atopic dermatitis extent and severity index (CADESI) of 25 were randomly allocated to receive PYM00217 (100, 200 or 400 mg kg-1 day-1) or placebo for 12 weeks. The mean reductions in CADESI (intention-to-treat population) were 3.9% (placebo; n=29), 4.4% (100 mg kg-1 day-1; n=30), 23.4% (200 mg kg-1 day-1; n=29) and 8.5% (400 mg kg-1 day-1; n=29). The reduction in the 200 mg kg-1 day-1 group was significant (P<0.01). For dogs with a baseline CADESI>or=50, the mean changes were +10.6% (placebo; n=12), +0.6% (100 mg kg-1 day-1; n=14), -29.3% (200 mg kg-1 day-1; n=14) and -3.4% (400 mg kg-1 day-1; n=15). The 200 mg kg-1 day-1 dose was significantly more effective than placebo (P=0.038). No serious adverse effects were reported. Minor adverse effects seen in 10% (placebo and 100 mg kg-1 day-1), 24% (200 mg kg-1 day-1) and 42% (400 mg kg-1 day-1) of cases were mainly minor gastrointestinal disorders and only five cases required cessation of dosing. Two dogs (one in each of the 100 mg kg-1 day-1 and 200 mg kg-1 day-1 groups) refused to eat the medicated food. In conclusion, PYM00217 at 200 mg kg-1 appears to be an effective, palatable and well-tolerated treatment for canine AD.

Animals↗

PubMed1998

Effect of Quillaja saponaria saponins and Yucca schidigera plant extract on growth of Escherichia coli.

Escherichia coli K-12 was exposed to Quillaja saponaria saponins from various commercial firms (Sigma, Roth and Nor-feed) and to an extract of Yucca schidigera plant powder (DK Sarsaponin 30) at different concentrations (0.05-1.0% w/v). A concentration-dependent response was observed. Quillaja saponaria saponins from Sigma increased growth up to 0.1% (w/v) level, whereas Nor-feed and Roth saponins produced maximum growth at a much higher level (0.5 and 0.75%, w/v, respectively). These results suggest that quillaja saponins from various sources differ in their biological activity, although all three saponins had the same content of vanillin-sulphuric acid reactive moieties. The lyophilized water extract from the DK Sarsaponin powder showed maximum growth at 0.1% (w/v) level. The levels at which maximum growth was observed did not change on subjecting the quillaja or yucca saponins to heat treatment in an autoclave (121 degrees C for 30 min). All the saponins and the plant extract increased growth of Escherichia coli up to a certain concentration and thereafter decreased growth. In spite of the decreased growth at higher levels of saponins, it was higher compared to the control (without saponin) up to levels of 1% (w/v) for all saponins except Quillaja saponins from Sigma, for which the growth was lower at levels of 0.25% (w/v) and higher. Saponins have the potential to modulate microbial growth in natural and artificial fermenters.

Anti-Bacterial Agents↗

PubMed2006

Detection and quantification of unbound phytochelatin 2 in plant extracts of Brassica napus grown with different levels of mercury.

The mercury (Hg) accumulation mechanism was studied in rape (Brassica napus) plants grown under a Hg concentration gradient (0 microm-1,000 microm). Hg mainly accumulated in roots. Therefore, the presence of phytochelatins (PCs) was studied in the roots of the plants. The high stability of the PC-Hg multicomplexes (mPC-nHg) seems to be the main reason for the lack of previous Hg-PC characterization studies. We propose a modification of the method to detect and quantify unbound PC of Hg in plant extracts via high-performance liquid chromatography coupled to electrospray tandem mass spectrometry and inductively coupled plasma mass spectrometry in parallel. We separated the PC from the Hg by adding the chelating agent sodium 2,3-dimercaptopropanesulfonate monohydrate. We only detected the presence of PC after the addition of the chelating agent. Some multicomplexes mPC-nHg could be formed but, due to their large sizes, could not be detected. In this study, only PC(2) was observed in plant samples. Hg accumulation was correlated with PC(2) concentration (r(2) = 0.98).

Brassica napus↗

PubMed2004

Investigation of the antimutagenic effects of selected South African medicinal plant extracts.

Dichloromethane extracts from different parts of Rhamnus prinoides, Ornithogalum longibracteatum, Gardenia volkensii, Spirostachys africana, Diospyros whyteana, Syzigium cordatum and Prunus africana were investigated for mutagenic and antimutagenic effects in Salmonella/microsome and micronucleus tests. None of the extracts tested in the Ames test were found to induce mutations or to modify the effect of the mutagen 4-nitroquinoline-oxide (4NQO). In the micronucleus test, extracts from twigs/bark of R. prinoides, twigs of D. whyteana, P. africana and S. cordatum significantly lowered the effect of the mutagen mitomycin C (MMC). Extracts from twigs/bark of G. volkensii and S. africana were genotoxic in the micronucleus test, while extracts of O. longibracteatum leaves potentiated the genotoxicity of MMC. This preliminary investigation shows that plant extracts used in traditional medicine may have particular effects with regard to mutagenicity and antimutagenicity indicating careful use in some instances and the need to isolate their active principles for further research.

4-Nitroquinoline-1-oxide↗

PubMed2005

RP-HPLC analysis of the phenolic compounds of plant extracts. investigation of their antioxidant capacity and antimicrobial activity.

Extracts of aromatic plants of Greek origin were examined as potential sources of phenolic compounds. RP-HPLC with UV detection was employed for the identification and quantification of the phenolic antioxidants, present in methanolic extracts. The most abundant phenolic acids were ferulic acid (1.1-280 mg/100 g of dry sample) and caffeic acid (1.2-60 mg/100 g of dry sample). (+)-Catechin and quercetin were the most abundant flavonoids. Apigenin and luteolin were detected in high amounts in Menta pulegium and Thymus vulgaris, respectively. The antioxidant capacity was determined, in dried ground plants and in their methanol extracts, with the Rancimat test using sunflower oil as substrate. Both pulverized plants and extracts showed antioxidant capacity. Total phenolic content in the extracts was determined spectrometrically according to the Folin-Ciocalteu assay and ranged from 1 to 21 mg of gallic acid/100 g of dry sample. Antimicrobial activity of the extracts against selected microbes was also conducted in this study.

Anti-Bacterial Agents↗

PubMed2000

Antifungal activity of plant extracts against Arthrinium sacchari and Chaetomium funicola.

Various plant extracts were examined for antifungal activity with the objective of improving the commercial sterility of aseptically filled tea beverage products in PET bottles. When the hot water extract and the methanol extract of 29 samples were measured for their antifungal activity against Arthrinium sacchari M001 and Chaetomium funicola M002 strains, five samples, Acer nikoense, Glycyrrhiza glabra, Lagerstroemia speciosa, Psidium guajava and Thea sinensis, showed high activity. Of these, the extracts from A. nikoense, G. glabra and T. sinensis were fractionated by extraction with CHCl3, and the CHCl3-soluble fractions from G. glabra showed antifungal activity with minimum inhibitory concentrations (MICs) between 62.5 and 125 microg/ml against the above-mentioned two fungi. When the EtOAc-soluble fraction of A. nikoense was used, the MIC against A. sacchari M001 was 62.5 microg/ml. However, none of the fractions from A. nikoense or T. sinensis showed high activity against C. funicola M002 and their MICs were greater than 500 microg/ml. A licorice preparation made from the commercially available oil-based extract of G. glabra showed a low MIC of 25 microg/ml against five tested strains of filamentous fungi, but not against Aspergillus fumigatus M008, in a blended tea. Consequently, the possibility of adding a licorice preparation made from the oil-based extract of G. glabra to tea beverages (aseptically filled into PET bottles) was suggested.

Journal Article↗

PubMedDate not supplied

[Antimutagenic activity of plant extracts from Armoracia rusticana, Ficus carica and Zea mays and peroxidase in eukaryotic cells].

Antimutagene activity and high efficiency of antimutagene action of plant extracts from horseradish roots (Armoracia rusticana), fig brunches (Ficus carica) and mays seedlings (Zea mays) and their ability to decrease the frequency of spontaneous and induced by gamma-rays chromosome aberrations in meristematic cells of Vicia faba and marrow cells of mice have been shown. Comparative assessment of genoprotective properties of peroxidase and the studied extracts has revealed higher efficiency of antimutagene action of peroxidase.

Animals↗

PubMed2005

Background correction in near-infrared spectra of plant extracts by orthogonal signal correction.

In near-infrared (NIR) analysis of plant extracts, excessive background often exists in near-infrared spectra. The detection of active constituents is difficult because of excessive background, and correction of this problem remains difficult. In this work, the orthogonal signal correction (OSC) method was used to correct excessive background. The method was also compared with several classical background correction methods, such as offset correction, multiplicative scatter correction (MSC), standard normal variate (SNV) transformation, de-trending (DT), first derivative, second derivative and wavelet methods. A simulated dataset and a real NIR spectral dataset were used to test the efficiency of different background correction methods. The results showed that OSC is the only effective method for correcting excessive background.

Algorithms↗

PubMed2005

[Controlling effects of plant extracts and pesticides on Myzus persicae and Lipaphis erysimi populations].

This paper studied the controlling effects of Xanthium sibiricum and Syngonium podophyllum ethanol extracts, mobile oil emulsion and 0.3% azadirachtin EC on Myzus persicae and Lipaphis erysimi populations both in laboratory and in field. The results showed that in laboratory, X. sibiricum extract was the key deterrent to Myzus persicae, while sprayed it with S. podophyllum extract and with petroleum spray oil or 0.3 % azadirachtin EC, the deterrent effect could be improved. As for pterigota Lipaphis erysimi, the extracts of the two plants were the main deterrents, and their interference effect was non-linear. In field, S. podophyllum extract had a stronger deterrent effect when sprayed independently. L. erysimi population could be successfully controlled by the mixtures of plant extracts and insecticides, with an efficiency of 95.7%. The best treatment against M. persicae population was X. sibiricum extract, with an efficiency of 87%, while S. podophyllum extract mixed with 0.3% azadirachtin EC, and X. sibiricum extract mixed with 0.3% azadirachtin EC had an efficiency of >80%.

Animals↗

PubMed2003

Protein glycation inhibitory and antioxidative activities of some plant extracts in vitro.

The protein glycation inhibitory activity of aqueous ethanolic extracts from 25 plant tissues was evaluated in vitro using the model system of bovine serum albumin and fructose. The most bioactive plant tissue was Allium cepa (skin), followed by Illicium religiosum (bark and wood), Fagopyrum esculentum (hull), Origanum officinalis (leaf), Rosmarinus officinalis (leaf), Pyrus pyrifolia (bark),Acanthopanax senticosus (bark), Eugenia caryophllata (leaf), and Erigeron annuus (whole). The extracts with glycation inhibitory activity also showed antioxidative activity when a micellar linoleic acid peroxidation system was applied followed by 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) radical cation decolorization and 1,1-diphenyl-2-picrylhydrazyl free radical scavenging assays. The glycation inhibitory activity was significantly correlated with the antioxidative potency of the extracts. The positive glycation inhibitory and antioxidative activities of these plants might suggest a possible role in targeting aging and diabetic complications.

Antioxidants↗