Effects of garlic powders with varying alliin contents on hepatic drug metabolizing enzymes in rats

The anticarcinogenic effect of garlic has been demonstrated in both epidemiologic and experimental studies. In this study, possible mechanisms involved in the anticarcinogenic effect of garlic consumption were assessed by determining its capacity to alter drug metabolizing enzymes, in relation with its alliin content. Rats were fed a diet for 2 weeks containing 5% garlic powders produced from bulbs grown on soils with different levels of sulfate fertilization and therefore containing differing amounts of alliin. Activities of several hepatic enzymes, which are important in carcinogen metabolism such cytochromes P450 (CYP) and phase II enzymes, were determined. Garlic consumption slightly increased ethoxyresorufin O-deethylase and CYP 1A2 levels. In contrast, garlic consumption decreased CYP 2E1 activity and the level of the corresponding isoform. UDP glucuronosyl transferase and glutathion S-transferase activities were increased by garlic powders. The alliin content of the garlic powders was positively correlated with UGT activity although not with other activities. Effects produced by garlic consumption were qualitatively similar to that of diallyl disulfide, a sulfur compound that has been extensively studied. These data could partially explain the chemoprotective effect of garlic.     

Composition, stability, and bioavailability of garlic products used in a clinical trial

In support of a new clinical trial designed to compare the effects of crushed fresh garlic and two types of garlic supplement tablets (enteric-coated dried fresh garlic and dried aged garlic extract) on serum lipids, the three garlic products have been characterized for (a) composition (14 sulfur and 2 non-sulfur compounds), (b) stability of suspected active compounds, and (c) availability of allyl thiosulfinates (mainly allicin) under both simulated gastrointestinal (tablet dissolution) conditions and in vivo. The allyl thiosulfinates of blended fresh garlic were stable for at least 2 years when stored at -80 degrees C. The dissolution release of thiosulfinates from the enteric-coated garlic tablets was found to be >95%. The bioavailability of allyl thiosulfinates from these tablets, measured as breath allyl methyl sulfide, was found to be complete and equivalent to that of crushed fresh garlic. S-Allylcysteine was stable for 12 months at ambient temperature. The stability of the suspected active compounds under the conditions of the study and the bioavailability of allyl thiosulfinates from the dried garlic supplement have validated the use of these preparations for comparison in a clinical trial.     

Differential effects of garlic oil and its three major organosulfur components on the hepatic detoxification system in rats.

The objective of this study was to compare the modulatory effect of garlic oil and its three organosulfur compounds, diallyl sulfide (DAS), diallyl disulfide (DADS), and diallyl trisulfide (DATS), on rat hepatic detoxification enzyme activity, and protein and mRNA expression. Rats were orally administered garlic oil (80 or 200 mg/kg bw), DAS (20 or 80 mg/kg bw), DADS (80 mg/kg bw), or DATS (70 mg/kg bw) three times a week for 6 weeks. Control rats received corn oil. According to the results, garlic oil and DAS in dosages of 200 and 80 mg/kg bw, respectively, significantly increased pentoxyresorufin O-dealkylase (PROD) activity as compared with the that of the control rats (P < 0.05). In contrast, N-nitrosodimethylamine demethylase activity in rats that received DADS and DATS was significantly lower than that in the control rats (P < 0.05). Ethoxyresorufin O-deethylase and erythromycin demethylase activities were not influenced by garlic oil, DAS, DADS, or DATS. To the phase II enzyme, garlic oil, DADS, and DATS significantly increased the glutathione S-transferase (GST) activity toward ethacrynic aicd (P < 0.05). Immunoblot assay showed that the protein contents of cytochrome P450 1A1, 2B1, and 3A1 were increased by garlic oil and each of three allyl sulfides, and the change among the allyl sulfides was in the order of DAS > DADS > DATS. The placental form of GST (PGST) level was also increased by garlic oil and the three allyl sulfides, but the increase among the allyl sulfides was DATS congruent with DADS > DAS. P450 2E1, however, was suppressed by each garlic component. Northern blot results indicated that the changes in P450 1A1, 2B1, 3A1, and PGST mRNA levels by garlic components were similar to those noted in the protein levels. These results indicate that the modulatory effect of garlic oil on hepatic drug-metabolizing enzymes can be attributed to its three major allyl sulfide components DAS, DADS, and DATS. These three allyl sulfides vary in modulatory activity, and this variation is related to the number of sulfur atoms in the molecule.     

Allicin and allicin-derived garlic compounds increase breath acetone through allyl methyl sulfide: use in measuring allicin bioavailability

Progress in establishing systemic pharmacological effects for fresh, crushed garlic (Allium sativum L) in humans has been hindered by (1) the inability to measure allicin bioavailability, (2) lack of direct evidence that allicin has significant systemic activity at doses of garlic normally consumed, and (3) lack of a model for an acute effect. We have addressed these problems by quantifying the increases in breath acetone and breath allyl methyl sulfide (AMS). The area under the 48 h curve was measured in humans after consumption of standardized garlic preparations, allicin, and allicin-derived compounds, at the equivalent of 7 g of crushed garlic. It was shown that the allyl thiosulfinates (mainly allicin) are solely responsible for breath AMS and increased breath acetone. Diallyl trisulfide, diallyl disulfide, ajoene, and S-allylmercaptocysteine, at isomolar dithioallyl, showed the same quantitative effects as allicin. Consumption of AMS at isomolar allyl also gave the same effects as allicin, indicating that AMS is the main metabolite of allicin and is an active metabolite. In conclusion, allicin and allicin-derived compounds are rapidly metabolized to AMS, a compound which stimulates the production of acetone and which can be used to measure the bioavailability of allicin and, hence, the ability of garlic supplements to represent fresh garlic.     

Diallyl disulfide and diallyl trisulfide up-regulate the expression of the pi class of glutathione S-transferase via an AP-1-dependent pathway

Garlic organosulfur compounds are recognized as potential chemopreventive compounds. This protection is related to the induction of phase II detoxification enzymes. We previously reported that diallyl disulfide (DADS) and diallyl trisulfide (DATS) up-regulate the gene expression of the pi class of glutathione S-transferase (GSTP) and that an enhancer element named GPE I is required for this induction. In the present study, we further investigated the signal pathway involved in DADS and DATS up-regulation of this detoxification enzyme in Clone 9 cells. Cells were cultured with 25-200 micromol/L of DADS or DATS for 24 h. Western and Northern blots showed that both garlic allyl sulfides concentration dependently induced GSTP protein and mRNA expression, respectively. Changes in GST activity toward ethacrynic acid were consistent with the increase in GSTP expression (P < 0.05). Electromobility gel shift assay showed that the DNA binding activity of nuclear activator protein-1 (AP-1) is concentration-dependently increased in the presence of DADS and DATS as compared with that of the control cells. The phosphorylation of c-Jun NH2-terminal kinase (JNK) and extracellular signal-regulated kinase (ERK), but not of p38, was stimulated in the presence of both garlic allyl sulfides. Pretreatment with SP600125 and PD98059, which are JNK and ERK inhibitors, respectively, abolished the increase in AP-1-DNA binding activity and also the induction of GSTP protein by either allyl sulfide. Our results indicate that the effectiveness of DADS and DATS on GSTP expression is likely related to the JNK-AP-1 and ERK-AP-1 signaling pathways and, thus, that DADS and DATS enhance the binding of AP-1 to GPE I.     

Induction of phase II detoxification enzymes in rats by plant-derived isothiocyanates: comparison of allyl isothiocyanate with sulforaphane and related compounds

Plants of the family Brassicaceae contain high levels of glucosinolates. The latter compounds are degraded to isothiocyanates, some of which have been shown to be potent inducers of phase II detoxification enzymes in vitro. In the present study, the ability of six plant-derived isothiocyanates (allyl isothiocyanate, iberverin, erucin, sulforaphane, iberin, and cheirolin) to increase tissue levels of the phase II detoxification enzymes quinone reductase (QR) and glutathione S-transferase (GST) in a variety of rat tissues has been compared. At the low dose level employed (40 micromol/kg/day), cheirolin was without effect in any tissue. All of the other isothiocyanates, however, increased GST and QR activities in the duodenum, forestomach, and/or the urinary bladder of the animals, with the greatest effects being seen in the urinary bladder. With the exception of cheirolin, little difference was observed in the inductive activity of the various isothiocyanates. Phase II enzymes are known to protect against chemical carcinogenesis, and the selectivity of isothiocyanates in inducing such enzymes in the bladder is of interest in view of recent epidemiological studies showing a decreased incidence of cancer of this organ in individuals with a high dietary intake of Brassica vegetables.     

Garlic allyl sulfides display differential modulation of rat cytochrome P450 2B1 and the placental form glutathione S-transferase in various organs

To investigate whether the regulation of garlic allyl sulfides on biotransformation enzyme expression is tissue-specific, the expression of cytochrome P450 2B1 (CYP 2B1) and the placental form of glutathione S-transferase (PGST) in liver, lung, and intestine, which are the three major organs responsible for drug metabolism, was examined. Rats were orally administrated 0.5 or 2 mmol/kg BW diallyl sulfide (DAS) or 0.5 mmol/kg BW diallyl disulfide (DADS) or diallyl trisulfide (DATS) three times per week for 6 weeks. The final body weights and the body weight ratio of liver and lung were not changed by any of these three allyl sulfide treatments as compared to the control rats. An 11- and 12-fold increase of 7-pentoxyresorufin O-dealkylase (PROD) activities was noted in rats treated with 0.5 or 2 mmol/mg BW DAS, respectively, as compared with the controls (P < 0.05). In contrast, DADS and DATS significantly increased hepatic PGST activity toward ethacrynic acid by 30 and 40%, respectively, as compared with the control rats (P < 0.05). An increase in PGST activity was only noted at 2 mmol/kg BW DAS group (P < 0.05). In addition, similar increases in PGST activity due to DADS and DATS were also noted in lung and jejunum tissue (P < 0.05). Immunoblot assay shows that the changes in CYP 2B1 and PGST proteins due to the three garlic allyl sulfide treatments on liver, lung, and jejunum were consistent with those observed for PROD and PGST activities. Northern blot further revealed that the DADS and DATS increased PGST mRNA levels in both liver (2.9- and 3.0-fold, respectively) and lung (4.1- and 2.6-fold, respectively) and DAS dose-dependently increased CYP 2B1 mRNA levels in the liver. Garlic allyl sulfides differentially induced CYP 2B1 and PGST expression, and this up-regulation of these two biotransformation enzymes is tissue-specific.     

Aqueous extract from Spanish black radish (Raphanus sativus L. Var. niger) induces detoxification enzymes in the HepG2 human hepatoma cell line

Spanish black radish (Raphanus sativus L. var. niger) is a member of the Cruciferae family that also contains broccoli and Brussels sprouts, well-known to contain health-promoting constituents. Spanish black radishes (SBR) contain high concentrations of a glucosinolate unique to the radish family, glucoraphasatin, which represents >65% of the total glucosinolates present in SBR. The metabolites of glucosinolates, such as isothiocyanates, are implicated in health promotion, although it is unclear whether glucosinolates themselves elicit a similar response. The crude aqueous extract from 0.3 to 3 mg of dry SBR material increased the activity of the phase II detoxification enzyme quinone reductase in the human hepatoma HepG2 cell line with a maximal effect at a concentration of 1 mg/mL. Treatment of HepG2 cells with the crude aqueous extract of 1 mg of SBR per mL also significantly induced the expression of mRNA corresponding to the phase I detoxification enzymes: cytochrome P450 (CYP) 1A1, CYP1A2, and CYP1B1 as well as the phase II detoxification enzymes: quinone reductase, heme oxygenase 1, and thioredoxin reductase 1. Previous studies have shown that the myrosinase metabolites of different glucosinolates vary in their ability to induce detoxification enzymes. Here, we show that while glucoraphasatin addition was ineffective, the isothiocyanate metabolite of glucoraphasatin, 4-methylthio-3-butenyl isothiocyanate (MIBITC), significantly induced phase II detoxification enzymes at a concentration of 10 microM. These data demonstrate that the crude aqueous extract of SBR and the isothiocyanate metabolite of glucoraphasatin, MIBITC, are potent inducers of detoxification enzymes in the HepG2 cell line.     

适宜的氮硫配施提高基质栽培大蒜鳞茎品质

试验采用蛭石-珍珠岩盆栽方式,研究了不同浓度氮(5、10、20 mmol/L,记作N1、N2、N3)、硫(2、4、8 mmol/L,记作S1、S2、S3)配施条件对大蒜鳞茎品质的影响,并采用隶属函数法对大蒜品质进行综合评价,分析鳞茎品质与氮硫互作水平的响应关系。试验结果表明,氮素、硫素对大蒜主要营养成分含量影响不尽相同,氮硫配施能够不同程度提高大蒜鳞茎中可溶性蛋白、VC、大蒜多糖、游离氨基酸的含量,且氮、硫单一因素对大蒜鳞茎品质影响远低于元素间交互作用。但在氮素适宜水平下(10~20 mmol/L),硫素水平对大蒜鳞茎中大蒜辣素含量有显著影响,提高硫浓度能够显著增加大蒜辣素含量,而在硫素浓度过高时(8 mmol/L)则明显抑制该物质合成。隶属函数法综合评价以N3S2处理组大蒜鳞茎品质最优,隶属函数值为0.81,对照组(CK)最差。综合分析可见,无机基质栽培条件下氮硫配施显著影响大蒜鳞茎品质,氮素浓度20 mmol/L(N3)、硫素浓度4 mmol/L(S2)时最利于大蒜鳞茎品质的提升,为大蒜无机基质栽培最佳氮、硫配施水平,且两元素配施存在明显的互作效应。该结论为大蒜栽培氮、硫元素合理配施提供试验指导。     

Storage life of field-grown garlic bulbs (Allium sativum L.) as influenced by nitrogen and sulfur fertilization

The most important active compound in garlic is alliin. Sulfur (S) fertilization was shown to significantly increase the alliin concentration in garlic cloves, while high nitrogen (N) levels had an adverse effect. The effect of graded N and S application on the storage life of garlic has been paid little attention so far. A bifactorial field trial with 4 levels of N and S was conducted in a randomized block design. At harvest, 40 bulbs per treatment were stored under terms comparable to the storage conditions in average households (20 °C, dry, and dim) for 83 days. Every 3 weeks, samples were analyzed for their alliin and water content. The alliin concentration in peeled garlic cloves increased during storage from on average 9.2 mg g(-1) dry weight at harvest to 21.4 mg g(-1) dry weight after 83 days of storage. S fertilization increased the alliin concentration by a factor of 2.3 from 11.4 mg g(-1) in the control treatment to 26.6 mg g(-1) dry weight at the highest S level of 45 kg ha(-1) after 83 days of storage. N fertilization decreased by a trend of the alliin content. Fertilizer rates had only a minor influence on water losses from bulbs at short-term storage. After 83 days of storage, water losses were by trend lower at higher S levels, and this relationship proved to be significant when no N was applied. Best quality in terms of high alliin contents was obtained during the entire storage time at an S level of at minimum 30 kg ha(-1) S if no N was applied. The results show that the physiological S demand of 15 kg ha(-1) S for optimum yield is lower than the S requirement of 30 kg ha(-1) S for a longer storage life.     

Protective Role of Selenium on Garlic Growth under Cadmium Stress

The stimulative action and detoxifying effect of selenium on garlic growth under cadmium stress were investigated. Selenite treatment can promote mitosis of garlic root-tip cells and growth of roots and seedlings. Garlic has a strong ability to accumulate the selenium from soil, and selenium content was greatest in garlic seedlings. Addition of selenite at a concentration of 2.0 μg kg^?1 in soil could decrease cadmium content in garlic and protect the seedling growth. The selenium has been shown to have an effective protective role on garlic growth under cadmium stress.     

Modulation of cytokine secretion by garlic oil derivatives is associated with suppressed nitric oxide production in stimulated macrophages

We previously described that garlic oil derivatives differentially suppress the production of nitric oxide (NO) and prostaglandin E(2) (PGE(2)) in activated macrophages. In the present study, we investigated the effects of the garlic derivatives, diallyl sulfide (DAS), diallyl disulfide (DADS), and allyl methyl sulfide (AMS), on cytokine production in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells, and the association between modulation of cytokines and inhibition of NO production was also assessed. The results indicated that these garlic compounds had different effects on the secretion of activated cytokines, including proinflammatory tumor necrosis factor-alpha (TNF-alpha), interleukin (IL)-1beta, and IL-6, as well as the antiinflammatory, IL-10. DAS inhibited the production of all stimulated cytokines in a concentration-dependent manner, and the inhibition was closely associated with the suppression of NO and PGE(2) production. DADS repressed the production of stimulated TNF-alpha and IL-10 and increased the production of activated IL-1beta and, to a lesser extent, IL-6; but only the decreased IL-10 production was associated with DADS-induced NO inhibition. Yet, the DAS- and DADS-suppressed NO production was independent of TNF-alpha. AMS, on the other hand, slightly suppressed the stimulated TNF-alpha but enhanced IL-10 production, and such modulation was closely associated with the decrease in NO production.     

Model studies on precursor system generating blue pigment in onion and garlic

Reactions involved in blue-green discoloration in a mixture of onion (Allium cepa L.) and garlic (Allium sativum L.) were investigated. Vivid-blue color was successfully reproduced by using a defined model reaction system comprising only trans-(+)-S-(1-propenyl)-L-cysteine sulfoxide (1-PeCSO) from onion, S-allyl-L-cysteine sulfoxide (2-PeCSO) from garlic, purified alliinase (EC 4.4.1.4), and glycine (or some other amino acids). Four reaction steps identified and factors affecting the blue color formation were in good agreement with those suggested by earlier investigators. When crude onion alliinase was used in place of garlic alliinase, less pigment was formed. This result was explained by a difference in the amount of thiosulfinates, colorless intermediates termed color developers, yielded from 1-PeCSO by these enzymes.     

Grapefruit and oroblanco enhance hepatic detoxification enzymes in rats: possible role in protection against chemical carcinogenesis

Citrus fruits are considered to be functional foods that promote good health. This study was carried out to assess the effect of oroblanco and grapefruit consumption on hepatic detoxification enzymes. Male Sprague-Dawley rats were provided with either regular drinking water (control) or experimental treatments of oroblanco juice, grapefruit juice, or a sugar mix for 6 weeks. After 1 week of treatment, half the animals in each group were injected with the procarcinogen 1,2-dimethylhydrazine. Grapefruit juice significantly increased activity and expression of the hepatic phase I enzyme, cytochrome P450 CYP1A1, with a marked trend toward enhanced NAD(P)H:quinone reductase (QR) activity. Oroblanco juice significantly increased glutathione S-transferase phase II enzyme activity along with CYP1A1 expression and a notable trend toward increased activity of both CYP1A1 and QR. These results suggest that these citrus fruits are bifunctional inducers, modulating both phase I and phase II drug-metabolizing enzymes to enhance hepatic detoxification.     

Low allicin release from garlic supplements: a major problem due to the sensitivities of alliinase activity

Most garlic supplements are standardized on allicin potential and are enteric-coated to prevent gastric acid inactivation of the allicin-producing enzyme, alliinase. To determine whether these products release the claimed amount of allicin under simulated gastrointestinal conditions, USP dissolution method 724A for drug release was applied to all 24 known brands of enteric-coated tablets. It was found that nearly all brands employed effective coatings and that they met their claims for allicin potential when crushed and suspended in water. However, all brands except one gave low dissolution allicin release, with 83% of the brands releasing less than 15% of their potential. The low allicin release was found to be due to both impaired alliinase activity, mostly caused by tablet excipients, and to slow tablet disintegration, which also impairs alliinase activity. Only when tablets had high alliinase activity and disintegrated rapidly did they show high allicin release. The ability of USP 724A to estimate allicin release in vivo was validated by monitoring breath levels of the allicin metabolite, allyl methyl sulfide. In conclusion, garlic powder supplements should no longer be standardized on allicin potential, but rather on dissolution allicin release.     

行星轮式大蒜插播机播种直立度优化与试验

大蒜机械化播种的植入环节中,在蒜种-土壤-触土部件强耦合作用下,正头后的蒜种直立度极易变低,如何“保姿植入”成为亟待解决的关键技术。针对此问题,该文以行星轮式大蒜插播机为研究对象,对插播鸭嘴的尖部运动轨迹进行分析,明晰了影响植后蒜种直立度的关键因素为插播鸭嘴的线速度、开启相位角及插播鸭嘴张开角度与凸轮凸起段对应的圆心角之比(开口速比)。运用Box-Benhnken中心组合试验方法对插播鸭嘴的线速度、开启相位角、开口速比进行三因素三水平二次回归试验设计,进行了插播试验,采用Design-expert软件建立响应面数学模型,对影响直立度的关键参数进行了综合优化,求解出最优工作参数组合为插播鸭嘴的线速度200 mm/s,开启相位角20°,开口速比2。大田试验结果表明,最优参数作业的蒜种直立度均值为63.2°,较优化前提高了21.8%,满足大蒜种植的蒜种直立度要求。     

Silicon alleviates mercury toxicity in garlic plants

Abstract

Silicon (Si) can increase plant stress tolerance. Mercury (Hg) is one of the major elements of heavy metal pollution. However, little attention has been paid to the possible effect of Si on Hg toxicity in plants. Here, the effects of Si on growth, photosynthesis, Hg accumulation and antioxidant defense were investigated in garlic grown in pots under Hg stress. Before sowing, Hg and Si were added at 3?mg Kg^?1 and 500?mg Kg^?1, respectively. The treatments included CT (control), Si, Hg and Hg?+?Si. The results showed that in non-stress conditions, added Si did not affect the garlic growth, photosynthetic gas exchange, malonaldehyde concentration or activities of antioxidant enzymes in leaves, except that it increased the superoxide dismutase activity. Under Hg stress, the garlic growth, leaf net photosynthetic rate, stomatal conductance, transpirational rate and superoxide dismutase activity in leaves were all inhibited, while the malondialdehyde concentration was increased; whereas these changes were all reversed in the presence of added Si. Added Si significantly decreased Hg concentrations in the root, bulb and shoot, and it also decreased exchangeable Hg level in the soil. These results suggest that Si could alleviate Hg toxicity in garlic through improving antioxidant defense ability, and decreasing Hg availability in soil and thus Hg uptake.     

The Role of Iron Bacteria on Weathering and Attenuation Processes at Acidic Environments

The present research reproduces the chemical and microbiological reactions that occur naturally when a metal sulfide is discharged onto a natural soil, with special emphasis on iron cycle. The role of indigenous microbiota from an extremely acidic site on both weathering and attenuation processes related to the iron mobilization has been studied and the iron cycle has been reproduced at laboratory scale. In the first stage, the weathering phase, a residual sulfide mineral was bioleached using a mixed culture of iron-oxidizing bacteria isolated from the own substrate. The acid liquor obtained (pH 2), with a high metal concentration (160 mM in total iron), was filtered and neutralized. Solids obtained from the two sources (from the weathering process and after the neutralization stage) were characterized by X-ray and scanning electron microscope/energy dispersive X-ray spectroscopy, resulting ferric iron precipitates such as jarosites, goethites, and ferrihydrites with different crystalline properties. The contribution of ferric iron-reducing bacteria on the attenuation of high-content iron effluents was also studied. Mixed cultures of ferric iron-reducing bacteria, isolated from those acidic substrates, were active in reducing soluble ferric iron (60 mM in concentration), and a 66% of bioreduction was reached after 15 days. Dissimilatory ferric iron reduction has been achieved with adapted cultures at pH values from 7 to 4.     

Isolation, characterization, and biological activity of naphthoquinones from Calceolaria andina L

Two compounds recognized as responsible for the insecticidal activity of extracts of Calceolaria andina L. (Scrophulariaceae) have been isolated and characterized as 2-(1, 1-dimethylprop-2-enyl)-3-hydroxy-1,4-naphthoquinone and the corresponding acetate, 2-acetoxy-3-(1,1-dimethylprop-2-enyl)-1, 4-naphthoquinone. Their activities against 29 pest species and 9 beneficial species of arthropod from a total of 11 orders have been determined. Activities against homopteran and acarine species are of the same order as those of established pesticides, and, significantly, no cross-resistance is observed for strains resistant to established classes of insecticide. Mammalian toxicities are low.