Prevention of arsenic-induced hepatic apoptosis by concomitant administration of garlic extracts in mice

Garlic is well known as a folk remedy for a variety of ailments since ancient times, however, very few studies are available suggesting its beneficial role against arsenic toxicity pertaining to its ability to eliminate arsenic from the blood and soft tissues and in reversal of arsenic-induced oxidative stress in affected tissues. The present study was planned to investigate the protective efficacy of aqueous garlic extract using two different doses on parameters suggestive of hepatic injury, tissue oxidative stress and mobilization of arsenic. Further, an attempt to understand the mechanism of arsenic in inducing hepatic apoptosis was also studied. Results of the present study suggested that arsenic administration in mice caused generation of reactive oxygen species (ROS) causing apoptosis through mitochondria-mediated pathway. The ROS generation in hepatic tissue reverted to normal values after co-administration of garlic extracts. The study provides significant evidence that garlic extracts contain strong anti-oxidant property which could be beneficial in preventing arsenic-induced toxicity in cells. However, further research is required to determine whether the results from animal studies are applicable to humans before garlic can be recommended as a putative agent against arsenic toxicity.     

Effect of application of different concentrations and appropriate schedules of aqueous garlic (Allium sativum L.) bulb extracts against Maruca vitrata and Clavigralla tomentosicollis on cowpea,Vigna unguiculata (L.) Walp

Abstract

Three concentrations (5, 10 and 20% w/v) and spraying schedules (2×, 4× and 6× weekly applications) of garlic, Allium sativum L. bulb extracts were assayed for efficacy against Maruca vitrata Fab. and Clavigralla tomentosicollis Stal. on cowpea under rain fed conditions in two seasons. The results showed that application of aqueous garlic bulb extracts at all the treatment levels significantly (p < 0.05) reduced the populations of the target pests compared with the untreated check. Application of 10 and 20% garlic bulb extracts with 4 or 6 weekly applications gave better control of these pests, protected the pods from damage and increased grain yields on treated cowpea compared with 5% extract at all three application schedules and were comparable to the synthetic insecticide treatment. This material could play a role in the integrated management of M. vitrata and C. tomentosicollis on cowpea fields managed by limited resource farmers in developing countries. The plant material is readily available, cheap, safe, non phytotoxic and technologically simple to use.     

P66Shc mediated ferritin degradation--a novel mechanism of ROS formation

Diallyl trisulfide (DATS) has been shown to induce the formation of reactive oxygen species (ROS) in prostate cancer cells, which was accompanied by a decrease in the ferritin protein level and an increase in the labile iron pool (LIP). However, the mechanism of the ferritin degradation has not been fully elucidated. In this paper we demonstrate that DATS-induced ROS formation depends on p66Shc. In cells stably expressing a dominant negative mutant of p66Shc (p66ShcS36A), DATS did not induce ROS formation. In addition, in cells expressing p66ShcS36A neither an increase in ferritin H degradation nor an increase in LIP were observed. Cells stably expressing p66ShcS36A also possess higher levels of ferritin H compared to PC-3 cells transfected with an empty vector. Moreover, DATS-induced G2/M arrest is completely abrogated in cells expressing p66ShcS36A. Mouse embryonic fibroblasts (MEFs) derived from wild-type (WT) or p66Shc knockout mouse have been used to evaluate if p66Shc involvement in DATS-induced signaling is cell specific. DATS induced G2/M arrest in WT MEFs but had no effect in the p66Shc^−/− cell line. Moreover, increases in LIP and ROS formation were significantly attenuated in p66Shc^−/− MEFs treated with DATS.     

蒜辣素UPLC检测体系优化及其在大蒜资源评价中的应用

大蒜是重要的调味蔬菜,大蒜辣素的含量是评价大蒜品质的最重要的一项指标。本研究通过对样品处理、提取方法和检测方法的研究,建立了完善的大蒜辣素超高效液相色谱(UPLC)检测方法:使用UPLC BEH C18色谱柱,流动相为甲醇∶水=1∶1,检测波长为254nm,进样体积为1μl,流速为0.3ml/min。大蒜辣素在2.04～510mg/L范围内呈良好线性关系(R2=0.9991)。利用建立的方法对212份大蒜鳞茎的大蒜辣素含量进行检测,发现供试212份大蒜资源的大蒜辣素含量差异显著,含量分布在0.82%～3.01%之间,最高含量与最低含量之间相差近4倍。     

Influence of gibberellic acid on auxin biosynthesis and their effects on coleoptile elongation in garlic

To demonstrate the effect of auxin on intact coleoptile growth, garlic (Allium sativum L.) cloves were inoculated in agar supplemented with DW (control), GA₃ and GA₃+tryptophan (a precursor of IAA, GA₃+T). The coleoptiles were harvested at 24 h intervals to measure growth in terms of length, activities of IAAld DH (which convert tryptophan to IAA) and peroxidase (that oxidizes IAA). Contents of endogenous IAA and PAA were also measured by indirect ELISA. Peroxidase activity was suppressed by GA₃ treatment and increased by GA₃+T treatment. Although endogenous contents of IAA were increased by the addition of GA₃ and even more by GA₃+T in the media, there was no further increase in coleoptile length, suggesting that garlic coleoptiles are sufficient in their production of IAA.     

A spectrophotometric assay for allicin,alliin, and alliinase (alliin lyase) with a chromogenic thiol: reaction of 4-mercaptopyridine with thiosulfinates

Allicin (diallylthiosulfinate) is the best known active compound of garlic. It is generated upon the interaction of the nonprotein amino acid alliin with the enzyme alliinase (alliin lyase, EC 4.4.1.4). Previously, we described a simple spectrophotometric assay for the determination of allicin and alliinase activity, based on the reaction between 2-nitro-5-thiobenzoate (NTB) and allicin. This reagent is not commercially available and must be synthesized. In this paper we describe the quantitative analysis of alliin and allicin, as well as of alliinase activity with 4-mercaptopyridine (4-MP), a commercially available chromogenic thiol. The assay is based on the reaction of 4-MP (lambda(max)=324nm) with the activated disulfide bond of thiosulfinates -S(O)-S-, forming the mixed disulfide, 4-allylmercaptothiopyridine, which has no absorbance at this region. The structure of 4-allylmercaptothiopyridine was confirmed by mass spectrometry. The method was used for the determination of alliin and allicin concentrations in their pure form as well as of alliin and total thiosulfinates concentrations in crude garlic preparations and garlic-derived products, at micromolar concentrations. The 4-MP assay is an easy, sensitive, fast, noncostly, and highly efficient throughput assay of allicin, alliin, and alliinase in garlic preparations.     

Garlic (Allium sativum L.) as a Potential Antidote for Cadmium and Lead Intoxication: Cadmium and Lead Distribution and Analysis in Different Mice Organs

Analysis and distribution of Pb and Cd in different mice organs including liver, kidney, spleen, heart and blood were evaluated after treatment with different aqueous concentrations of garlic (12.5–100 mg/l). Atomic absorption spectrometry (AAS) was used for analysis of Pb and Cd in these organs. Treatment of Cd–Pb exposed mice with garlic (12.5–100 mg/l) reduced Pb concentrations by 44.65, 42.61, 38.4, 47.56, and 66.62% in liver, kidney, heart, spleen and blood respectively. Moreover, garlic reduced Cd levels by 72.5, 87.7, 92.6, 95.6, and 71.7% in liver, kidney, heart, spleen and blood respectively. The suppressed immune responses in mice pretreated with Cd–Pb mixture were reversed by 48.85, 55.82, 81.4 and 90.7 in the presence of 100, 50, 25, and 12.5 mg/ml of garlic extract.     

Comparative insecticidal activity of different plant materials from six common plant species against Tribolium castaneum (Herbst) (Coleoptera: Tenebrionidae)

Tribolium castaneum (Herbst) (Coleoptera: Tenebrionidae) is one of the major insect pests of stored grains. Due to export legislation and zero-tolerance for live insect in trade commodities, extensive use of synthetic insecticides is in practice in order to eliminate pest infestations from the lots. Currently, the one and only acceptable chemical to be used in stored grain is phosphine but due to its excessive usage the stored grain pests are becoming resistant against it. Hence discovery of alternative compounds is much needed. In this study we have compared insecticidal efficacy of different plant materials from six commonly grown plants of Pakistan, viz. Allium sativum (Alliaceae), Zingiber officinale (Zingiberaceae), Cymbopogon citratus (Poaceae), Eucalyptus globulus (Myrtaceae), Nicotiana tabacum (Solanaceae), and Azadirachta indica (Meliaceae) against T. castaneum infesting stored wheat, rice, corn and gram pulse. Various plant parts were dried, powdered, and used as admixtures to the stored commodities in the experiments. The results have suggested that A. sativum (garlic) and Z. officinale (ginger) were more effective resulting into 15 times higher adult mortality and 4 to 5 times reduction in grain weight losses when mixed with rice grains. Similarly, A. indica when admixture with wheat checked the population growth in the resources resulting into 3.5 times less adult production compared to controls. A subsequent experiment was conducted to study the dose response of neem seed powder against the beetle pest infesting milled products. Surprisingly, better control was observed either at lowest (1% w/w) or the highest doses (5% w/w). This finding is of great interest to understand the underlying phenomenon which we assume is the ability of T. castaneum to feed selectively in flour mediums, however, further research on this aspect is required to be investigated. The results of this study support the use of botanicals for stored product pest management.     

Studies on the Silver-Staining of Plant Chromosomes

In the rye, barley and wheat, besides the Ag-NORs, some other Ag-stained regions including the telomeres, the centromeres and the middle of arms have been found. The phenomenon that the rye and barley have their own Ag-staining pattern is first described in this paper.     

大蒜多糖对中毒性心肌炎心肌细胞凋亡的影响

目的:研究大蒜多糖 (GP)对中毒性心肌炎心肌细胞凋亡及Bcl -2、Bax蛋白表达的影响,探讨GP抑制心肌细胞凋亡的可能机制。方法:建立小鼠阿霉素 (ADR)中毒性心肌炎模型,利用缺口末端标记法检测心肌凋亡细胞;免疫组化法检测心肌细胞bcl 2、bax基因的蛋白表达情况,并利用电镜观察心肌结构变化。结果:ADR( 3mgkg-1ip,qod× 7)可致小鼠心肌细胞凋亡数明显升高,心肌细胞线粒体水肿,促进心肌细胞凋亡蛋白bax和抑制细胞凋亡蛋白bcl 2含量均明显升高,但bax/ bcl- 2的比值同时升高明显,与正常组比较有显著差异性 (P <0 . 0 1 )。GP可逆转ADR所致的上述改变,表现为剂量依赖性抑制细胞凋亡,降低bax蛋白表达同时增加bcl 2表达,使bcl- 2/ bax的比值增加 (P <0 .0 5或P <0 . 0 1 )。结论:GP能拮抗阿霉素所致的小鼠中毒性心肌炎心肌细胞凋亡作用,其作用机制可能与抑制Bax基因的蛋白表达,使Bcl -2基因表达的蛋白功能相对增强,Bcl -2 /Bax比值升高有关。