Polymorphisms of the DNA repair gene XRCC1 and the frequency of somatic mutations at the glycophorin A locus in newborns

Essential oils extracted from the three medicinal plants; Helichrysum italicum, Ledum groenlandicum and Ravensara aromatica, together with their mixture were tested for their genotoxic and antigenotoxic activities against urethane, a well-known promutagen. We have adopted the somatic mutations and recombination test (SMART) in the wings of Drosophila melanogaster. Three days old larvae, trans-heterozygous for two genetic markers mwh and flr, were treated by essential oil and/or urethane. A negative control corresponding to solvent was also used. Our results do not show any significant effect of the oils tested but they reduce the mutation ratio resulting from urethane. The mixture of the three oils at equal volume seems to be the most effective. The antimutagenic effect of these oils could be explained by the interaction of their constituents with cytochrome P-450 activation system leading to a reduction of the formation of the active metabolite. The effect could also be attributed to certain molecules that are involved in these oils.     

Genotoxicity and anti-genotoxicity of some traditional medicinal herbs

Six herbal infusions used worldwide (Matricaria chamomilla, Tilia cordata, Mentha piperita, Mentha pulegium, Uncaria tomentosa and Valeriana officinalis) were assayed for anti-genotoxicity using the Somatic Mutation And Recombination Test (SMART) in Drosophila melanogaster. All these infusions are traditionally used for various medical purposes, including anti-inflammatory processes. Hydrogen peroxide was used as an oxidative genotoxicant to test the anti-genotoxic potency of the medicinal infusions. None of these infusions showed a significant genotoxicity, quite the reverse they were able to behave as desmutagens, detoxifying the mutagen hydrogen peroxide. The phenolic content of such herbal infusions is argued to be the possible scavenger of reactive oxygen radicals produced by the hydrogen peroxide.     

Modulatory effects of Duguetia furfuracea (A. St. Hil) Benth. and Hook. f. in Drosophila melanogaster somatic and germinative cells

Mutagenic and antimutagenic activities of the medicinal plant Duguetia furfuracea were assessed using SMART/wing and ring-X-loss tests. For the ring-X-loss test, 2- to 3-day-old Drosophila melanogaster ring-X-lineage males and virgin ywsn3 females received D. furfuracea infusion at doses of 0.085, 0.042, or 0.014 g/mL for 24 h. We found that D. furfuracea did not produce any mutagenic effects in D. melanogaster germinative cells. The somatic cells of D. melanogaster were analyzed using the SMART/wing test involving three lineages - mwh, flr3, and ORR - and the same doses of D. furfuracea infusion employed in the ring-X-loss test, as well as 20 mM urethane. The results of both standard (ST) and high bioactivation (HB) crosses showed absence of mutagenic activity of D. furfuracea. In contrast, in both ST and HB crosses, we observed a modulatory effect of D. furfuracea against the genotoxic activity of urethane.     

Genotoxicity and antigenotoxicity of some essential oils evaluated by wing spot test of Drosophila melanogaster

Essential oils extracted from the three medicinal plants; Helichrysum italicum, Ledum groenlandicum and Ravensara aromatica, together with their mixture were tested for their genotoxic and antigenotoxic activities against urethane, a well-known promutagen. We have adopted the somatic mutations and recombination test (SMART) in the wings of Drosophila melanogaster. Three days old larvae, trans-heterozygous for two genetic markers mwh and flr, were treated by essential oil and/or urethane. A negative control corresponding to solvent was also used. Our results do not show any significant effect of the oils tested but they reduce the mutation ratio resulting from urethane. The mixture of the three oils at equal volume seems to be the most effective. The antimutagenic effect of these oils could be explained by the interaction of their constituents with cytochrome P-450 activation system leading to a reduction of the formation of the active metabolite. The effect could also be attributed to certain molecules that are involved in these oils.     

The genotoxic and anti-genotoxic effects of <Emphasis Type="Italic">Stachys petrokosmos</Emphasis> leaf extract in human lymphocytes using microsomal fractions

The genotoxic and anti-genotoxic effects of Stachys petrokosmos leaf extracts (Sp) were investigated in human lymphocytes. The cells were treated with 1.5, 3.0 and 6.0 μL/mL concentrations of Sp leaf extracts for 24 and 48 h treatment periods in the absence and presence of metabolic activator (S9mix). In the absence of S9mix, Sp alone did not induce chromosome aberrations and formation of micronucleus while inducing the mean sister chromatid exchange at the highest concentration. In addition, Sp decreased the mutagenic effect of mitomycin-c. Sp alone showed a cytotoxic effect determined by a decrease in the proliferation index, mitotic index and nuclear division index. On the other hand a mixture of Sp and mitomycin-c resulted in a higher cytotoxic effect especially for 48 h treatment period. In the presence of S9mix, Sp was not genotoxic and cytotoxic however, it showed an anti-genotoxic effect by decreasing the effects of cyclophosphamide.     

High-Throughput In Vivo Genotoxicity Testing: An Automated Readout System for the Somatic Mutation and Recombination Test (SMART)

Genotoxicity testing is an important component of toxicity assessment. As illustrated by the European registration, evaluation, authorization, and restriction of chemicals (REACH) directive, it concerns all the chemicals used in industry. The commonly used in vivo mammalian tests appear to be ill adapted to tackle the large compound sets involved, due to throughput, cost, and ethical issues. The somatic mutation and recombination test (SMART) represents a more scalable alternative, since it uses Drosophila, which develops faster and requires less infrastructure. Despite these advantages, the manual scoring of the hairs on Drosophila wings required for the SMART limits its usage. To overcome this limitation, we have developed an automated SMART readout. It consists of automated imaging, followed by an image analysis pipeline that measures individual wing genotoxicity scores. Finally, we have developed a wing score-based dose-dependency approach that can provide genotoxicity profiles. We have validated our method using 6 compounds, obtaining profiles almost identical to those obtained from manual measures, even for low-genotoxicity compounds such as urethane. The automated SMART, with its faster and more reliable readout, fulfills the need for a high-throughput in vivo test. The flexible imaging strategy we describe and the analysis tools we provide should facilitate the optimization and dissemination of our methods.     

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.     

Genotoxic evaluation of selective serotonin-reuptake inhibitors by use of the somatic mutation and recombination test in Drosophila melanogaster

This study evaluated different concentrations of selective serotonin-reuptake inhibitors (citalopram and sertraline) for genotoxicity by use of the somatic mutation and recombination test (SMART) in Drosophila melanogaster. Three-day-old larvae, trans-heterozygous for the multiple wing hairs (mwh) and flare (flr3) genes were treated with these two compounds. Two recessive markers were located on the left arm of chromosome 3, i.e. 'multiple wing hairs' (mwh) in map position 0.3 and 'flare-3' (flr3) at 38.8, while the centromere was located in position 47.7. SMART is based on the loss of heterozygosity, which may occur through various mechanisms, such as mitotic recombination, mutation, deletion, half-translocation, chromosome loss, and non-disjunction. Genetic changes occurring in somatic cells of the wing's imaginal discs, cause the formation of mutant clones on the wing blade. The results of this study show that citalopram had a genotoxic effect in the Drosophila SMART. Sertraline, however, did not show any genotoxic effect in balancer heterozygous wings. This study concluded that more information is needed to be certain regarding the mutagenic effects of sertraline.     

THE EFFECT OF URETHANE ON THE CONSUMPTION OF OXYGEN AND THE RATE OF CELL DIVISION IN THE CILIATE TETRAHYMENA GELEII

1. The inhibition of oxygen consumption produced by a series of concentrations of ethyl carbamate has been measured in the protozoan Tetrahymena geleii. 2. The relation found between the narcotic concentration and its effect on respiration leads to the conclusion that urethane has two distinct modes of action in this cell. The respiratory data can be accurately predicted by assuming that the inhibitor acts on two independent parallel respiratory systems. 3. Complete suppression of cell division in this organism is brought about by approximately 0.1 M urethane. 4. Urethane concentrations up to 0.1 M affect primarily only one of the two postulated respiratory systems. The mechanism of the narcosis of cell division in this organism by urethane thus appears to be inhibition of this "activity" system.     

Investigation of the mutagenic and antimutagenic effects of Origanum compactum essential oil and some of its constituents

In the present study, the chemical composition of Origanum compactum essential oil was determined by gas chromatography and mass spectrometry, and its mutagenic and antimutagenic activities were investigated by the somatic mutation and recombination test (SMART) in Drosophila melanogaster. No significant increase in the number of somatic mutations was observed with the essential oil tested using both the standard (ST) and high bio-activation (HB) cross. In order to investigate the antimutagenic effect of the essential oil, we have tested the effect on the indirect-acting mutagen urethane (URE), as well as the direct-acting mutagen methyl methanesulfonate (MMS). O. compactum essential oil showed a strong inhibitory effect against URE-induced mutagenicity, especially with the HB cross. However, only a weak inhibitory effect on the mutagenicity induced by MMS was observed. These results suggest that the detected antimutagenicity could be mediated by an inhibitory effect on metabolic activation. The essential oil was fractionated to identify the components responsible of the suppressing effect detected. Seven fractions were obtained: two of them showed the most potent inhibitory effect against URE-induced mutagenicity and were further fractionated. The sub-fractions obtained from the second chromatographic fractionation were tested for their antimutagenic activity, together with carvacrol and thymol. The highest antimutagenic effect obtained with the sub-fractions was similar to the effect of the crude essential oil, as well as to the effect of carvacrol alone. These results suggest the absence of a synergic antimutagenic effect between the components of O. compactum essential oil and indicate that carvacrol was the most active oil component.     

Influence of solid state fermentation by Trichoderma spp. on solubility,phenolic content,antioxidant, and antimicrobial activities of commercial turmeric

The influence of solid state fermentation (SSF) by Trichoderma spp. on the solubility, total phenolic content, antioxidant, and antibacterial activities of turmeric was determined and compared with unfermented turmeric. The solubility of turmeric was monitored by increase in its phenolic content. The total phenolic content of turmeric extracted by 80% methanol and water after SSF by six species of Trichoderma spp. increased significantly from 2.5 to 11.3–23.3 and from 0.5 to 13.5–20.4 GAE/g DW, respectively. The antioxidant activities of fermented turmeric were enhanced using 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2′-azino-bis (3-ethylbenzo-thiazoline-6-sulfonic acid) (ABTS), and ferric ion-reducing antioxidant power (FRAP) assays. The antibacterial activity of fermented turmeric against human-pathogenic bacteria Escherichia coli, Streptococcus agalactiae, Staphylococcus aureus, Entreococcus faecalis, Methicillin-Resistant S. aureus, Klebsiella pneumonia, and Pseudomonas aeruginosae showed a broad spectrum inhibitory effect. In conclusion, the results indicated the potentials of using fermented turmeric as natural antioxidant and antimicrobial material for food applications.     

Pharmacological basis for the use of turmeric in gastrointestinal and respiratory disorders

This study was carried out to provide scientific basis for the medicinal use of turmeric (Curcuma longa) in gastrointestinal and respiratory disorders. The crude extract of turmeric (Cl.Cr), relaxed the spontaneous and K+ (80 mM)-induced contractions in isolated rabbit jejunum as well as shifted the CaCl2 concentration-response curves. In rabbit tracheal preparation, Cl.Cr inhibited carbachol and K(+)-induced contractions. In anesthetized rats, Cl.Cr produced variable responses on blood pressure with a mixture of weak hypertensive and hypotensive actions. In rabbit aorta, Cl.Cr caused a weak vasoconstrictor and a vasodilator effect on K+ and phenylephrine-induced contractions. In guinea-pig atria, Cl.Cr inhibited spontaneous rate and force of contractions at 14-24 times higher concentrations. Activity directed fractionation revealed that the vasodilator and vasoconstrictor activities are widely distributed in the plant with no clear separation into the polar or non-polar fractions. When used for comparison, both curcumin and verapamil caused similar inhibitory effects in all smooth muscle preparations with relatively more effect against K(+)-induced contractions and that both were devoid of any vasoconstrictor effect and curcumin had no effect on atria. These data suggest that the inhibitory effects of Cl.Cr are mediated primarily through calcium channel blockade, though additional mechanism cannot be ruled out and this study forms the basis for the traditional use of turmeric in hyperactive states of the gut and airways. Furthermore, curcumin, the main active principle, does not share all effects of turmeric.     

广西几个姜黄品种姜黄油GC-MS分析

通过对广西地区栽培的三个姜黄品种:博白姜黄、大新姜黄、市场姜黄的姜黄油GC-MS分析,比较不同姜黄品种姜黄油成分的差异性,为筛选特定姜黄品种提供依据。不同姜黄品种其主要成分含量差异显著,主成份姜黄酮(含芳香-姜黄酮)含量依次为博白姜黄56.07%、大新姜黄47.79%、市场姜黄28.38%,市场姜黄的姜黄酮含量只有博白姜黄的一半。因此从姜黄油主要成份(含量>20%)姜黄酮含量来考察,以种植博白姜黄为最好。     

Effect of pentobarbital or urethane on bile secretion and chemical composition of blood in the rabbit

A study was made of the effects of anaesthesia with pentobarbital and urethane on the bile secretion and the chemical composition of the blood of New Zealand rabbits. Neither of the agents was observed to affect arterial pH or pO2, but with urethane pCO2 values decreased significantly. This was associated with a pronounced hyperglycaemia. Bile flow was significantly higher in pentobarbital-anaesthetized animals than in urethane-anaesthetized animals, a phenomenon that can be attributed to a different canalicular flow and that will not be related to differences in the bile-acid-dependent fraction of secretion. Under both anaesthetics, bile sodium concentrations were greater than those found in plasma, which can be explained by the formation of micelles with low osmotic activity. Bile bicarbonate concentrations proved to be greater than those observed in plasma. Plasma calcium concentrations were significantly lower with urethane than with pentobarbital, whereas in bile the situation was reversed; both these aspects are discussed.     

Chemical composition of turmeric oil--a byproduct from turmeric oleoresin industry and its inhibitory activity against different fungi

Curcumin, the yellow coloring pigment of turmeric is produced industrially from turmeric oleoresin. The mother liquor after isolation of curcumin from oleoresin known as curcumin removed turmeric oleoresin (CRTO) was extracted three times with n-hexane at room temperature for 30 min to obtain turmeric oil. The turmeric oil was subjected to fractional distillation under vacuum to get two fractions. These fractions were tested for antifugal activity against Aspergillus flavus, A. parasiticus, Fusarium moniliforme and Penicillium digitatum by spore germination method. Fraction II was found to be more active. The chemical constituents of turmeric oil, fraction I and fraction II were determined by GC and identified by GC-MS. Aromatic turmerone, turmerone and curlone were major compounds present in fraction II along with other oxygenated compounds.     

Evaluation of antioxidant activities and antimutagenicity of turmeric oil: a byproduct from curcumin production

Curcumin removed turmeric oleoresin (CRTO) was extracted with hexane concentrated to get turmeric oil, and that was fractionated using silica gel column chromatography to obtain three fractions. These fractions were analyzed by GC and GC-MS. Turmeric oil contained aromatic turmerone (31.32%), turmerone (15.08%) and curlone (9.7%), whereas fractions III has aromatic turmerone (44.5%), curlone (19.22%) and turmerone (10.88%) as major compounds Also, oxygenated compounds (5,6,8-10) were enriched in fraction III. Turmeric oil and its fractions were tested for antioxidant activity using the beta-carotene-linoleate model system and the phosphomolybdenum method. The fraction III showed maximum antioxidant capacity. These fractions were also used to determine their protective effect against the mutagenicity of sodium azide by means of the Ames test. All the fractions and turmeric oil exhibited a markedly antimutagenicity but fraction III was the most effective. The antioxidant effects of turmeric oil and its fractions may provide an explanation for their antimutagenic action.     

Genotoxicity of garlic, turmeric and asafoetida in mice

The genotoxic effects of orally administered garlic and turmeric were evaluated in bone-marrow cells of mice by performing the micronucleus test. Another spice, asafoetida, was tested for the induction of sister-chromatid exchanges (SCEs) in spermatogonia of mice. Results of the micronucleus test with garlic and turmeric were not significantly different from control values. Orally administered asafoetida, however, showed a weak SCE-inducing effect in spermatogonia.     

乌拉坦对大鼠海马CA1神经元电压门控钠通道和动作电位的作用

乌拉坦对兴奋性和抑制性配体门控通道具有广泛的可检测的作用.作者运用全细胞膜片钳技术研究乌拉坦对wistar大鼠海马CA1神经元电压门控钠通道和动作电位的作用.结果发现乌拉坦可逆并剂量依赖性地抑制钠电流和动作电位,其中,在10mmol/L浓度时可减小钠电流强度达38%,使激活曲线向去极化方向移动,并延长钠通道失活后的恢复时间,降低动作电位的幅值.这些结果表明乌拉坦对电压门控钠通道的抑制作用可能是乌拉坦全身麻醉作用的机制之一.     

Low doses of urethane effectively inhibit spinal seizures evoked by sudden cooling of toad isolated spinal cord.

The effect of low doses of urethane on three phases of spinal seizures evoked by sudden cooling (SSSC) of toad isolated spinal cord was studied. In control toads, SSSC began with a latency of 91 +/- 3 sec (mean +/- S.E.M.) exhibiting brief tremors, followed by clonic muscle contractions and finally reaching a tonic contraction (tonic phase). The latency of onset of seizures was significantly enhanced. The tonic phase was markedly abolished in toads pretreated intralymphatically with 0.15 g/kg of urethane. Tremors were the only phase observed in 55% of toads that received doses of 0.2 g/kg, and a total blockade of seizures was seen after doses of 0.25 g/kg of urethane in 50% of the preparations. A possible depressant effect of urethane on transmission mediated by excitatory amino acids is suggested.     

水合氯醛、乌拉坦及其1：1混合液在SD大鼠麻醉中的效果比较及应用

目的：比较水合氯醛、乌拉坦及其1：1混合液在SD大鼠麻醉中的效果并进一步在大鼠模型制备的麻醉中检验其效果。方法：分别采用不同剂量的水合氯醛和乌拉坦及其1：1混合液进行麻醉实验,比较其麻醉起效时间、维持时间和死亡率,并将相同剂量的1：1混合液应用于SD大鼠模型制作时的麻醉中,比较其与非模型组之间的差异。结果：水合氯醛和乌拉坦混合液麻醉大鼠的起效时间2.5±1.5分钟,与单用水合氯醛无差异（P〉0.05）,比单用乌拉坦起效时间短（P〈0.05）;维持时间107.4±4.1分钟,比单用水合氯醛、乌拉坦长（P〈0.01）;麻醉死亡率比单用水合氯醛低,总死亡率比单用水合氯醛、乌拉坦低。模型组大鼠的麻醉起效时间2.9±1.6分钟,维持时间108.9±4.4分钟,零麻醉死亡率,总死亡率为2.5%;与1：1混合液非模型组的麻醉效果没有明显差异。结论：水合氯醛＋乌拉坦1：1混合液麻醉效果好、起效快、死亡率极低,适合用于2小时左右的SD大鼠手术或模型制作。