The effect of turmeric extracts on inflammatory mediator production.

Major compounds of several commonly used botanicals, including turmeric, have been purported to have anti-inflammatory actions. In order to test the anti-inflammatory activity of compounds isolated from rhizomes of Curcuma longa L. (Zingiberaceae), we have established an in vitro test system. HL-60 cells were differentiated and exposed to lipopolysaccharide (LPS) from Escherichia coli (1 microg/ml) in the presence or absence of botanical compounds for 24 h. Supernatants were collected and analyzed for the production of tumor necrosis factor alpha (TNF-alpha) and prostaglandin E2 (PGE2) using standard ELISA assays. Water-soluble extracts were not cytotoxic and did not exhibit biological activity. Organic extracts of turmeric were cytotoxic only at concentrations above 50 microg/ml. Crude organic extracts of turmeric were capable of inhibiting LPS-induced TNF-alpha (IC50 value = 15.2 microg/ml) and PGE2 (IC50 value = 0.92 microg/ml) production. Purified curcumin was more active than either demethoxy- or bisdemethoxycurcumin. Fractions and subfractions of turmeric extracts collected via preparative HPLC had differing biological activity, ranging from no activity to IC50 values of < 1 microg/ml. For some fractions, subfractionation resulted in a loss of activity, indicating interaction of the compounds within the fraction to produce an anti-inflammatory effect. A combination of several of the fractions that contain the turmeric oils was more effective than the curcuminoids at inhibiting PGE2. While curcumin inhibited COX-2 expression, turmeric oils had no effect on levels of COX-2 mRNA.     

Allelopathic effect of Bromus spp. and Lolium spp. shoot extracts on some crops

Allelopathy is an untapped resource for weed control in crops that could give good possibilities for environmentally sound, integrated crop production. Allelopathy is defined as the direct or indirect harmful or beneficial effects of one plant on another through the production of chemical compounds, called allelochemicals, which escape into the environment. Allelochemicals can be produced by weeds and affect crops, and the reverse is also true. Allelopathic interactions include weed-weed, weed-crop, and crop-crop. Allelopathy offers potential for selective biological weed control for instance weed-suppressing crops and the use of plant residues in cropping systems, allelopathic rotational crops, or companion plants with allelopathic potential. Bromus species occur in many habitats in temperate regions of the world, including America, Eurasia, Australia, and Africa. The genus Lolium is one of the most important forage grasses. The weed species usually grow in the same production zones as wheat and are considered weeds since they parasitize wheat fields. Some of the weed species in these two genus have been reported to have allelopathic effect. One of the methods that has been successful in studying allelopathic activity are bioassays. Laboratory experiments were conducted to determine allelopathic effect of watery shoot extracts of four weed species of the Poaceae family, namely Bromus rigidus, Bromus diandrus, Lolium multiflorum and Lolium temulentum on germination and growth of winter wheat (Triticum aestivum L.), spring barley (Hordeum vulgare L.), corn (Zea mays L), perennial ryegrass (Lolium perenne L.), bean (Phaseolus sp.) and sunflower (Helianthus annuus L.) and on each other. The experiment was carried out during the period March 2010 to October 2010. Twenty five seeds were put into one Petri-dish on filter paper, adding 15ml of extract to each in four repeats. The germination took place in a Binder-type thermostat in the dark. The timing of germination was checked in every two days and the rate of growth was estimated after a week, by counting the number of germinated seeds and measuring the length of the radicle and plumule. The measured data was statistically analyzed and the effect of the extracts on germination percentage and seedling length was evaluated.     

Direct vasoactive and vasoprotective properties of anthocyanin-rich extracts.

Reactive oxygen species (ROS) play a critical role in the impairment of nitric oxide-mediated vascular functions and overall pathogenesis associated with cardiovascular disease. Plant pigment anthocyanins are exceptionally potent oxygen radical scavengers that produce beneficial effects in diseases outside the cardiovascular system. We examined for the first time the potential coronary vasoactive and vasoprotective properties of three anthocyanin enhanced extracts prepared from chokeberry (Ck), bilberry (B), or elderberry (E). Coronary arterial rings were isolated from 64 pigs and incubated in sterile tissue culture media overnight for use in one of four separate in vitro isometric force recording studies. Ck and B, but not E, produced dose- and endothelium-dependent vasorelaxation. (%maximal relaxation at 5 mg total anthocyanins per liter: Ck = 68 +/- 11, B = 59 +/- 10). Coronary vascular tone, endothelium-dependent vasorelaxation to A23187, and vasorelaxation to DEA NONOate were not affected by exposure of rings to any extract at 0.05 mg total anthocyanins per liter for 5 or 30 min. Ck extract at 0.05 mg total anthocyanins per liter showed the greatest protection against loss of A23187 relaxation following exposure to ROS from pyrogallol (Ck, % maximal relaxation and -logED50 to A23187, respectively, means +/- SE: Ck alone, 93 +/- 5%, 7.91 +/- 0.1; pyrogallol alone, 76 +/- 7%, 7.46 +/- 0.06; pyrogallol + Ck, 98 +/- 1%, 7.82 +/- 0.06; control: 99 +/- 1%, 7.86 +/- 0.07; P < 0.05 control vs. pyrogallol alone). Neither the extracts nor pyrogallol affected responses to DEA NONOate. Thus anthocyanin-enhanced extracts produce endothelium-dependent relaxation in porcine coronary arteries. Extract concentrations too low to directly alter coronary vascular tone protect coronary arteries from ROS without altering vasorelaxation to endogenous or exogenous NO. These results suggest that such extracts could have significant beneficial effects in vascular disease.     

Hepatocellular toxicity of kava leaf and root extracts.

Kava extracts are used widely for different purposes and were thought to be safe. Recently, several cases of hepatotoxicity have been published. To explore possible mechanisms of kava hepatotoxicity, we prepared and analyzed three different kava extracts (a methanolic and an acetonic root and a methanolic leaf extract), and investigated their toxicity on HepG2 cells and isolated rat liver mitochondria. All three extracts showed cytotoxicity starting at a concentration of 50 microg/ml (lactate dehydrogenase leakage) or 1 microg/ml (MTT test). The mitochondrial membrane potential was decreased (root extracts starting at 50 microg/ml) and the respiratory chain inhibited and uncoupled (root extracts) or only uncoupled (leaf extract) at 150 microg/ml, and mitochondrial beta-oxidation was inhibited by all extracts starting at 100 microg/ml. The ratio oxidized to reduced glutathione was increased in HepG2 cells, whereas the cellular ATP content was maintained. Induction of apoptosis was demonstrated by all extracts at a concentration of 150 microg/ml. These results indicate that the kava extracts are toxic to mitochondria, leading to inhibition of the respiratory chain, increased ROS production, a decrease in the mitochondrial membrane potential and eventually to apoptosis of exposed cells. In predisposed patients, mitochondrial toxicity of kava extract may explain hepatic adverse reactions of this drug.     

西芹种子子浸提液对黄瓜枯萎病菌的化感作用

为探讨西芹种子浸提液对黄瓜枯萎病菌是否有化感作用及其化感作用强度的大小,采用蒸馏水、乙醇、丙酮作为浸提剂,获得西芹种子的浸提液.用生长速率法研究其对黄瓜枯萎病菌的化感作用.结果表明:西芹种子的水、乙醇、丙酮浸提液都对黄瓜枯萎病菌有化感作用,浓度越大化感作用越强;50 mg·ml-1浓度的3种浸提液处理后144 h时与对照相比抑制黄瓜枯萎病菌蒸馏水浸提液为31.1%,乙醇浸提液为49.6%,丙酮浸提液为57.5%;乙醇、丙酮浸提液的化感作用强度大于蒸馏水浸提液.     

Triplex-induced recombination in human cell-free extracts. Dependence on XPA and HsRad51

Triple helix-forming oligonucleotides (TFOs) can bind to polypurine/polypyrimidine regions in DNA in a sequence-specific manner. Triple helix formation has been shown to stimulate recombination in mammalian cells in both episomal and chromosomal targets containing direct repeat sequences. Bifunctional oligonucleotides consisting of a recombination donor domain tethered to a TFO domain were found to mediate site-specific recombination in an intracellular SV40 vector target. To elucidate the mechanism of triplex-induced recombination, we have examined the ability of intermolecular triplexes to provoke recombination within plasmid substrates in human cell-free extracts. An assay for reversion of a point mutation in the supFG1 gene in the plasmid pSupFG1/G144C was established in which recombination in the extracts was detected upon transformation into indicator bacteria. A bifunctional oligonucleotide containing a 30-nucleotide TFO domain linked to a 40-nucleotide donor domain was found to mediate gene correction in vitro at a frequency of 46 x 10(-)5, at least 20-fold above background and over 4-fold greater than the donor segment alone. Physical linkage of the TFO to the donor was unnecessary, as co-mixture of separate TFO and donor segments also yielded elevated gene correction frequencies. When the recombination and repair proteins HsRad51 and XPA were depleted from the extracts using specific antibodies, the triplex-induced recombination was diminished, but was either partially or completely restored upon supplementation with the purified HsRad51 or XPA proteins, respectively. These results establish that triplex-induced, intermolecular recombination between plasmid targets and short fragments of homologous DNA can be detected in human cell extracts and that this process is dependent on both XPA and HsRad51.     

钮子七提取物的抗氧化作用及对胃癌细胞的抑制作用

以钮子七提取物为材料,通过邻苯三酚自氧化法测定钮子七提取物对超氧阴离子生成的影响,通过邻二氮菲溶液测定钮子七提取物对羟自由基的清除作用;采用噻唑蓝(MTT)法测定钮子七提取物对胃癌细胞生长的影响。结果显示,钮子七水提取物和70%乙醇提取物对超氧阴离子的产生有较强的抑制作用,在浓度大于5.0 mg/m L时,抑制作用显著增强。钮子七70%乙醇提取物和水提取物均有清除羟自由基的作用,在浓度大于2.5 mg/m L时清除作用显著增强。钮子七水提物和乙酸乙酯提取物在24 h、48 h及72 h后对胃癌细胞均有抑制作用,且都以作用72 h后抑制作用最强。结果表明,钮子七提取物具有较强的抗氧化作用以及较强的抑制胃癌细胞的作用。     

Effect of double-strand breaks on homologous recombination in mammalian cells and extracts.

We examined the effect of double-strand breaks on homologous recombination between two plasmids in human cells and in nuclear extracts prepared from human and rodent cells. Two pSV2neo plasmids containing nonreverting, nonoverlapping deletions were cotransfected into cells or incubated with cell extracts. Generation of intact neo genes was monitored by the ability of the DNA to confer G418r to cells or Neor to bacteria. We show that double-strand breaks at the sites of the deletions enhanced recombination frequency, whereas breaks outside the neo gene had no effect. Examination of the plasmids obtained from experiments involving the cell extracts revealed that gene conversion events play an important role in the generation of plasmids containing intact neo genes. Studies with plasmids carrying multiple polymorphic genetic markers revealed that markers located within 1,000 base pairs could be readily coconverted. The frequency of coconversion decreased with increasing distance between the markers. The plasmids we constructed along with the in vitro system should permit a detailed analysis of homologous recombinational events mediated by mammalian enzymes.     

Toxicity and pharmacology of extracts from dorid nudibranches.

1. Aqueous extracts of digestive glands of specimens of the dorid nudibranchs Cadlina flavomaculata, Doriopsilla albopunctata, Anisodoris nobilis, Archidoris montereyenis, and A. odhneri were lethal when injected into shore crabs and when injected intraperitoneally into mice. 2. Aqueous extracts of the degestive glands of Doriopsilla albopunctata and of Anisodoris nobilis were shown by bioassay (guinea pig ileum)and by chemical determination to contain histamine. The amount present was far too small to account for the toxicity of the glands. 3. Extracts of the digestive glands of Anisodoris nobilis were fractionated by column chromatography on Biogel P-2 to yield an active fraction designated "dorid toxin". This produces lethargy and bradycardia in mice. In anesthetized rats it produces sustained (60 min or more) bradycardia and hypotension. On isolated hearts, especially spontaneously beating guinea pig atria, it has negative inotropic and chronotropic effects. 4. Dorid toxin has a molecular weight under 8000. It is heat stable and is not destroyed by trypsin, chymotrypsin or Pronase. It is therefore unlikely that it is a polypeptide.     

Cysteine synthesis by Desulfovibrio vulgaris extracts.

Extracts of Desulfovibrio vulgaris were found to contain serine transacetylase and cysteine synthase activities. When extracts were incubated with bisulfite and o-acetylserine, or acetyl coenzyme A plus L-serine, under a hydrogen atmosphere, cysteine was formed. Pyruvate served as a reductant for bisulfite reduction to sulfide and concomitantly provided the acetyl moiety for acetyl coenzyme A formation. Consequently, when extracts were incubated with pyruvate, bisulfite, and L-serine, cysteine synthesis resulted.