Double mutation of Saccharomyces cerevisiae for enhanced β-D-fructofuranosidase fructohydrolase productivity and application of growth kinetics for parametric significance analysis

Abstract The kinetics of an extracellular β-D-fructofuranosidase fructohydrolase production by Saccharomyces cerevisiae in a chemically defined medium, i.e., sucrose peptone agar yeast extract at pH 6, was investigated. The wild-type was treated with a chemical mutagen, methyl methane sulfonate. Among the six mutants isolated, methyl methane sulfonate-V was found to be a better enzyme producing strain (52 ± 2.4a U/mL). The maximum production (74 ± 3.1a U/mL) was accomplished after at 48 h (68 ± 2.7a mg/mL protein). The mutants were stabilized at low levels of 5-fluoro-cytocine and the viable ones were further processed for optimization of cultural conditions and nutritional requirements. The sucrose concentration, incubation period and pH were optimized to be 30 g/L, 28 °C, and 6.5, respectively. The methyl methane sulfonate-V exhibited an improvement of over 10 folds in enzyme production when 5 g/L ammonium sulfate was used as a nitrogen source. Thin layer chromatography and high-performance liquid chromatography analysis illustrated the optimal enzyme activity supported by the higher rate of hydrolysis of sucrose into monosaccharides, particularly α-D-glucose and β-D-fructose. The values for Qp (2 ± 0.12c U/mL/h) and Yp/s (4 ± 1.24b U/g) of the mutant were considerably increased in comparison with other yeast strains (both isolates and viable mutants). The mutant could be exploited for enzyme production over a wider temperature range (26–34 °C), with significantly high enzyme activity (LSD 0.048, HS) at the optimal temperature.

Some biochemical and immunological changes associated with use of garlic extract [allicin] in combating some fish pathogens

We studied the effects of garlic [allicin] which used as immunostimulant on some physiological, biochemical parameters, survival rate, histopathological studies and bacteriological characteristics of sea bream. Fish [50 +/- 5 g/fish] were assigned to 3 treatments, with three replicates each. Treatment groups had a different level of allicin [Garlen[registered]], [1 and 2 gm. /kg feed] added to their diets; the control group diet was free from garlic. Diets also contained 25% crude protein [CP] and were administered at a rate of 3% live body weight twice daily for 8 weeks. At the end of 7[th] week ten fish from each treatment group and from the control were artificially infected by intra peritoneal injection with 0.2 ml of culture suspension of pathogenic Vibrio alginolyticus previously adjusted to 10[4]. Results showed that the differential leucocytic count, phagocytic activity, phagocytic index, serum lysozomal and bactericidal activity increased significantly with increasing levels of allicin. Plasma Cortisol level decreased significantly with increasing levels of allicin. Total protein and globulin were also increased significantly in the treated groups than the control group. All bacterial counts decreased significantly in treated groups than the control in all weeks. Treated groups had lower mortality rate than the control group during the challenge test. Histopathological studies of control group revealed congestion in blood vessel of gills, liver, spleen and kidney with, hyperplasia of epithelial cells between secondary lamellae led to fusion, edema and lifting of the lamellar epithelium of secondary lamellae in gills, vacuolar degeneration of hepatocytes, with infiltration by monocytes in liver, dilation of blood vessel, severe depletion of white bulb, edema, and hyperactivation of melanomacrophage center in spleen, necrosis of renal tubules epithelium, and inter renal haemopoietic tissues with atrophied glomerular tuft and widening of bowman capsules in kidney. Moderate to minimal histopathological changes appeared in all organs in groups received [Garlen[registered]] according to its dose

Experimental study on effect of allicin in inhibiting insulin-induced vascular smooth muscle cell proliferation and migration / 中国中药杂志

<p><b>OBJECTIVE</b>To investigate the effect of allicin in inhibiting insulin-induced vascular smooth muscle cell (VSMC) proliferation and migration.</p><p><b>METHOD</b>The tissue explant method was adopted to culture rat's aVSMCs, and the immunofluorescence method was used to identify α-SMA. Cells from the third to fifth generations were selected in the experiment The insulin-induced VSMC model was established. The experiment was carried out in five groups: the control group, the insulin group, the allicin group, the ERK inhibitors PD98059 group (20 μmol · L(-1)) and the PD98059 + allicin group. VSMCs' proliferation was determined by CCK8 colorimetric method, while its migration was detected by cell counting; The western blotting was used to detect total ERK, Phospho-ERK, PCNA protein's expression.</p><p><b>RESULT</b>Primary cultured VSMCs grew well in the spindle shape under the lightmicroscope, with peak and valley. α-SMA immunofluorescence results showed that the cultured cells had typical VSMCs' features. Insulin could stimulate VSMCs' proliferation and migration, with the best effect at the concentration of 100 nmol · L(-1). The pretreatment with allicin could significantly inhibit VSMCs' proliferation and migration induced by insulin in a dose-dependent manner. The pretreatment with PD98059 and allicin + PD98059 could inhibit VSMCs' proliferation and migration induced by insulin remarkably as well. Insulin could significantly accelerate VSMCs' expression of such proteins as p-ERK, PCNA. Contrarily, allicin could notably inhibit VSMCs' expression of such proteins as p-ERK, PCNA in a dose-dependent manner.</p><p><b>CONCLUSION</b>Allicin could significantly inhibit VSMCs' proliferation and migration induced by insulin, which may be related to the inhibition of the activation of ERK signal path.</p>

Antitumoral activity of allicin from garlic / Actividad antitumoral de alicina de ajo

Epidemiological studies link increased garlic consumption with a reduced incidence of cancer in various human populations. Experimental carcinogenesis studies in animal models and in cell culture systems indicate that several allium-derived compounds exhibit inhibitory effects. To provide a better understanding of the effects of allium derivatives on the prevention of cancer, we examined allicin, the major component of garlic, for their effects on antitumoral activity in vitro and in L5178Y lymphoma bearing mice. We found that allicin decreased the growth of tumor cells whereas in vivo, the compound shown an antitumor effect in murine L5178Y lymphoma. Allicin enhanced the secretion of IL-2, IFN-gamma and TNF-alpha cytokines from mouse plasma. These cytokines are associated with the beneficial Th1 antitumor response, which is characteristic of effective cancer immunotherapies.

Los estudios epidemiológicos relacionan el aumento del consumo de ajo con una disminución en la incidencia de cáncer en diferentes poblaciones humanas. Los estudios experimentales de carcinogénesis en modelos animales y en los cultivos de células tumorales indican que varios compuestos derivados del ajo tienen efectos inhibitorios. A fin de proporcionar una mejor comprensión de los efectos de derivados del ajo en la prevención del cáncer, se evaluó la alicina el principal componente del ajo, en la actividad antitumoral in vitro y en ratones con linfoma. Se encontró que la alicina disminuyó el crecimiento de células tumorales, mientras que in vivo, el compuesto muestra un efecto antitumoral en el linfoma murino L5178Y. La alicina incrementó la secreción de las citocinas IL-2, interferón-gamma y TNF-alfa obtenidas de plasma de ratón con linfoma. Estas citocinas están asociadas con la respuesta antitumoral benéfica Th1, que es característica de inmunoterapias efectivas para el cáncer.

Ultrastructural study on the morphological changes to male worms of Schistosoma mansoni after in vitro exposure to allicin / Estudo ultraestrutural das alterações morfológicas de vermes machos de Schistosoma mansoni após exposição in vitro à alicina

INTRODUCTION: Garlic has a wide range of actions, including antibacterial, antiviral, antifungal, antiprotozoal and anthelmintic actions. This antiparasitic activity has been attributed to allicin, which is the main constituent of garlic. The present study aimed to investigate the in vitro activity of allicin on the tegument of adult Schistosoma mansoni worms using scanning electron microscopy. METHODS: Swiss Webster mice were infected with S. mansoni cercariae (100 per mouse) and sacrificed 50 days later to acquire the adult worms. These worms were collected by perfusion and placed in RPMI medium 1,640 at 37°C before transferring to RPMI media containing 0 (control), 5, 10, 15 and 20mg/mL of allicin, where they were incubated for 2h. The worms were fixed in 2.5 percent glutaraldehyde solution, washed twice, post-fixed in osmium tetroxide, washed twice and then dehydrated with ascending grades of ethanol. The samples were air-dried, mounted on stubs, gold coated in an ion sputtering unit and viewed using a scanning electron microscope. RESULTS: A concentration of 5mg/mL caused wrinkling in the tegument; a concentration of 10mg/mL resulted in changes to tubercles and loss or modification of spines. With 15 and 20mg/mL increasing damage to the tegument could be seen, such as vesicle formation and the presence of ulcers. CONCLUSIONS: These findings demonstrate the effect of allicin on adult S. mansoni worms and indicate that most of the changes occur at concentrations greater than that normally indicated for treatment.

INTRODUÇÃO: O alho apresenta uma ampla gama de ações, incluindo antibacteriana, antiviral, antifúngico, antiprotozoário e anti-helmíntico. Esta atividade antiparasitária tem sido atribuída à alicina, que é o principal constituinte do alho. O presente estudo teve como objetivo investigar a ação in vitro da alicina no tegumento de vermes adultos de Schistosoma mansoni utilizando a microscopia eletrônica de varredura. MÉTODOS: Camundongos Swiss Webster foram infectados com cercárias de S. mansoni (100 por camundongo) e sacrificados 50 dias depois para aquisição de vermes adultos. Estes vermes foram coletados por perfusão e colocados em meio RPMI 1.640 a 37°C antes de transferir para o meio RPMI contendo 0 (controle), 5, 10, 15 e 20mg/mL de alicina, onde eles foram incubados por 2h. Os vermes foram fixados em uma solução de glutaraldeído a 2,5 por cento, lavados duas vezes, pós-fixados em tetróxido de ósmio, lavados duas vezes e então desidratados em séries crescentes de etanol. As amostras foram secadas, montadas em stubs, metalizadas em ouro e visualizadas utilizando o microscópio eletrônico de varredura. RESULTADOS: A concentração de 5mg/mL causou o enrugamento do tegumento; a concentração de 10mg/mL resultou em alterações nos tubérculos e perda ou modificações nos espinhos. Com 15 e 20mg/mL crescentes danos no tegumento pode ser visto, tais como formação de vesículas e presença de úlceras. CONCLUSÕES: Esses resultados demonstram os efeitos da alicina nos vermes adultos de S. mansoni e indicam que a maioria das alterações ocorrem numa concentração maior do que a normalmente indicada para o tratamento.

Protective effect of allicin on human periodontal ligament cells with nicotine-induced oxidative damage / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To explore the protective effect of allicin on nicotine-induced oxidative damage to human periodontal ligament cells (HPDLCs).</p><p><b>METHODS</b>(1) Establish nicotine-induced oxidative damage model on HPDLCs. Use water-soluble tetrazolium (WST) colorimetric method to find out the nicotine concentration (X) that could inhibit HPDLCs' growth for the following experiments. (2) HPDLCs of the fifth passage were divided into 5 groups: The control group, the nicotine group and the nicotine+allicin groups(the concentration of allicin was 15, 30, and 60 microg x mL(-1) respectively). Different kinds of culture media were added. Similarly, use WST colorimetric method to choose the allicin concentration (Y) that could significantly improve the survival rate of HPDLCs. (3) HPDLCs were divided into 3 groups: The control group, the nicotine group, the nicotine+allicin group and different media were added. The glutathion (GSH) concentrations in HPDLCs were determined in 1, 4, 8, 12 and 24h respectively.</p><p><b>RESULTS</b>0.8 mg x mL(-1) nicotine could inhibit the HPDLCs survival rate significantly (77% of the control, P < 0.05). But 60 microg x mL(-1) allicin could prevent the inhibition effects evidently, improving the survival rate to 112% of that of the nicotine group (P < 0.05) and reaching the survival rate level of control group (P > 0.05). The GSH concentrations of nicotine+allicin group were higher than that of the nicotine group always (P < 0.05) and by 82% at 8 h after culture, but had no difference with that of the control group (P > 0.05).</p><p><b>CONCLUSION</b>60 microg x mL(-1) allicin can protect the HPDLCs against oxidative damage induced by nicotine.</p>

Study on inhibitory effects of allicin in mucoepidermoid carcinoma cells / 临床耳鼻咽喉头颈外科杂志

OBJECTIVE@#To observe the anticancer mechanism of allicin by observing the inhibitory effect of allicin on human salivary gland carcinoma cell line MEC-1.@*METHOD@#Cell proliferation were measured by MTT assay at different doses and different hours. In the meantime, cell cycle was detected via flow cytometry after different dose incubation with different hours.@*RESULT@#MTT results showed that the inhibitory rates of MEC-1 proliferations were increased in a concentration-and time-dependent manner. Flow cytometry analysis showed percent-age of MEC-1 cells decreased in G0/G1 phase and increased in G2/M. But there was no evident change in S phase. The cells were mainly blocked in M phase, and the inhibitory effect of the allicin on MEC-1 cells increased with the increasing of concentration and time.@*CONCLUSION@#Allicin can inhibit the growth of MEC-1 cells in vitro dramatically.

Cardioprotective effect of garlic juice on the isolated rat heart in ischemia-reperfusion

It has been reported that the chronic oral administration of garlic homogenate protected the rat heart from in vitro ischemic reperfusion injury. However, the biological effects of garlic juice on the heart are expected to be different from oral administration of it. The present study was designed to investigate the effect of garlic juice on the isolated rat heart in ischemia- reperfusion. Rat isolated, perfused hearts were subjected to 30 min baseline measurement followed by 40 min normothermic global ischemia and 45 min reperfusion. Garlic juice [0.01 mg/ml] was added to the perfusion solution 20 min before ischemia in the test 1 and 5 min before and 10 min after ischemia in test 2. Different cardiac variables including left ventricular developed pressure [LVDP], heart rate [HR] and coronary flow [CF] were measured. Rate pressure product [RPP] was calculated, and released lactate dehydrogenase [LDH] enzyme in effluent was measured in reperfusion. Garlic juice significantly increased CF before ischemia in both test groups. The released LDH enzyme at the first minute and the recovery of RPP and LVDP on the 45[th] minute of reperfusion were significantly better in the test group 2 in comparison to the control. The result of the present study shows that garlic juice has a vasodilator activity and protects the isolated ischemic rat heart when it was administrated in reperfusion. It is probably mediated by antioxidant activity of allicin as a principal bioactive compound of garlic juice

Allicin enhances cytotoxicity of CPT-11 to colon cancer LoVo cell in vitro / 中国中药杂志

<p><b>OBJECTIVE</b>To study the effect of allicin on human colon cancer cell line LoVo and the combined effect of allicin and CPT-11 on this cancer cell line.</p><p><b>METHOD</b>The LoVo cells were cultured in vitro and treated with allicin in different concentrations. MTT assay was used to test dynamically the cell growth inhibiting effect. Apoptosis induction (Annexin-V-FITC/PI) and modulation of DNA cell cycle were measured by flow cytometry. The change of cytotoxicity of CPT-11 after combination of allicin at the concentration of 4.0, 8.0 mg x L(-1) were investigated.</p><p><b>RESULT</b>Allicin had inhibitive effect on growth of LoVo cells in a dose and time dependent manner, with IC50 value of 32.23, 10.74, 6.58 mg x L(-1) at 24 h, 48 h and 72 h, respectively. The apoptosis rate of LoVo cells increased progressively as the cells were treated with increasing concentration of allicin in 24 h, while the apoptosis rate achieved peak value when the cells were treated with allicin at the concentration of 8 mg x L(-1) in 48 h. The result indicated the low concentrations of allicin (< 4 mg x L(-1)) lead to G2/M cell cycle arrest, and higer concentrations ( > 4 mg x L(-1)) exert G1 + G2/M cell cycle arrest in 24 h. Compared with single use of CPT-11, the combined use of CPT-11 and allicin (4.0, 8.0 mg x L(-1), respectively) showed increasing cytotoxicity on the LoVo cells, with IC50 of 24 h decreasing from 47.5 to 7.4 and 7.2 mg x L(-1), respectively.</p><p><b>CONCLUSION</b>Allicin has significant anti-proliferation effect on human colon cancer cell line LoVo by induction of apoptosis and arrestment of cell cycle and can enhance the cytotoxicity of CPT-11 on the colon cancer LoVo cell.</p>

Metabonomic study on the effects of allicin on rats / 药学学报

To investigate the effects of allicin on rats by NMR-based metabonomic method, the changes of endogenous metabolites in normal rat urine and the influences on metabolism were analyzed with bio-nuclear magnetic resonance (NMR) method and partial least-squares discriminant analysis (PLS-DA) after intraperitoneal administration of allicin solution. The identified biochemical effects associated with allicin dosing included elevated then gradually recovered urinary levels of Kreb's cycle intermediates, such as citrate, alpha-ketoglutarate and succinate and increased concentrations of ketones. Meanwhile, decreased urinary concentrations of glucose, lactate, alanine, hippurate and trimethylamine oxide were observed. The PLS-DA revealed that the metabonomic profiles of allicin treated groups were obviously different from those of the control group. Allicin may change metabolism significantly in normal rats. The study of the pharmacologic mechanism of allicin by metabonomic method is practicable and it could be explored further.

Effect of Allicin of garlic on production nitric oxide of macrophage to Candida albicans

Garlic is an herbal plant having various compounds. one of the most important of which is allicin with antibiotic property. Candida Albicans is opportunistic yeast which in case of immune system dysfunction is considered as a pathogenic agent. Immune mechanisms against this fungus are Macrophages which act with oxidative and non-oxidative fungicide mechanisms. Oxidative mechanism includes active oxygen and nitrogen mediators which are produced by active macrophages and thereby the microorganisms are eliminated. The aim of this study was to investigate Allicin effects on the increase activites macrophage to product of Nitric Oxide It is examined in this study the effect of garlic allicin on macrophages' activity in releasing Nitric oxide against Candida Albicans. Garlic allicin was prepared by the method of chloroformic extract, and then made to react with macrophages from male mouse Balb/c with 2-8 week of age in vitro. Candida Albicans was divided into two groups: with and without allicin. Both cases compared as positive and negative samples, the rate of macrophages' activity was determined through production of Nitric Oxide. After examining the rate of Nitric Oxide produced by macrophages, results showed that allicin as a natural material activates immune system against this fungus, so that macrophages with allicin can produce more Nitric Oxide than the group without allicin. This was obtained by comparing the results from these two groups and the control group. Regarding the important role of Candida Albicans in Candidiasis and applying the preventing agents by this fungus in suppressing the macrophages' activities in producing Nitric Oxide, and also by studying related literatures, we concluded that allicin extracted from garlic can affect greatly on the macrophages' activities in producing Nitric Oxide against agents of Candidiasis disease

Effect of P38MAPK signal transduction pathway on apoptosis of THP-1 induced by allicin / 中国中药杂志

<p><b>OBJECTIVE</b>The objective of this paper was to study the change of P38MAPK and Fas in the apoptosis of THP-1 cells induced by allicin.</p><p><b>METHOD</b>The proliferation inhibition rates of THP-1 cells after various treatments were examined by MTT assay. Apoptosis rate was determined with Annexin V- FITC/PI double staining by flow cytometry. The expression and distribution change of the phosphorylation p38MAPK (P-p38MAPK) were detected by immunohistochemical staining. The changes of P-p38 MAPK and Fas proteins were detected by Western blot.</p><p><b>RESULT</b>The proliferations of leukemia cell line THP-1 are inhibited by allicin. MTT assay showed that allicin can inhibit the proliferation of the THP-1 cell, and the inhibition was dependent on both dose and time. The IC50 of 72 hours was 12.8 mg x L(-1). Apoptosis rate detected by Annexin V-FITC/PI was proportional to the concentration of the allicin. After the immunohistochemical staining test, the P-p38MAPK was located in the cell nucleus and plasma, showing deep brown, when adding allicin to THP-1 cell. Western blot test showed that the P-p38MAPK proteins expression was proportional to the concentration of Allicin and was also dose dependent. The levels of P-p38MAPK in negative control group, 1/2 IC50 of 72 hours group and IC50 of 72 hours group were 0.259 8 +/- 0.013 2, 0.61 2 +/- 0.008 3 and 0.505 6 +/- 0.005 5 respectively, and the levels of Fas proteins were 0.287 4 +/- 0.008 9, 0.426 8 +/- 0.007 9 and 0.597 1 +/- 0.010 9 respectively. The difference was statistically significant when compared with the negative control group (P < 0.01).</p><p><b>CONCLUSION</b>Allicin can significantly induce THP-1 cells apoptosis, and its mechanism may be related to the activation of P38MAPK/Fas.</p>

HPLC tandem-mass spectrometric analysis of the chemical components and decomposition products of allicin extract of garlic / 药学学报

To analyze the chemical components and decomposition products in allicin extract of garlic, the chemical components screening and identification were made with HPLC-MS/MS method by full scan TIC MS, HPLC retention time, product MS spectra and chemical reference standards. The stability of the extract in water and alcoholic solutions was also investigated. There were five major components in allicin extract which were all identified as thiosulfinates. The extract was stable for at least 3 months when stored at -20 degrees C as water solution, but obvious decomposition was observed with the increase of alcoholic concentration. The decomposition products were also identified by HPLC-MS/MS.

Effect of local application of allicinvia gastroscopy on cell proliferation and apoptosis of progressive gastric carcinoma / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To study the effects of local application of allicin via gastroscopy on progressive gastric carcinoma, and to investigate its possible mechanisms.</p><p><b>METHODS</b>Eighty patients with progressive gastric adenocarcinoma, whose diagnosis was confirmed by gastroscopy and pathological examination, were assigned to 2 groups, 40 in each group. Forty-eight hours before operation, allicin was infused via gastroscopy to the lesion region of patients in the allicin group, and normal saline was infused instead to those in the control group. The gastric carcinoma tissue gotten from gastrectomy was taken to determine the percentage of cells in various cell cycle phases ( G0/ G1, S and G2/M), the cell apoptosis rate, proliferation index value and apoptosis related gene protein such as Fas, Bax and Bcl-2 by flow cytometry.</p><p><b>RESULTS</b>In the allicin group, the cell apoptosis rate was 9.60 +/- 1.52%, the percentage of cell in G0/G1 phase was 72.12 +/- 8.35%, in G2/M phase 9.54 +/- 3.20%, and PI 27.80 +/- 8.35, while in the control group, the corresponding data was 2.20 +/- 0.58%, 69.56 +/- 5.15%, 13.20 +/- 3.05%, and 30.40 +/- 5.15, respectively, and significant difference in all the 4 indexes could be found between the two groups (P < 0.05, P < 0.01). Moreover, allicin showed effects in up-regulating the protein expressions of apoptosis promoting gene Bax and apoptosis initiating gene Fas (P < 0.05, P < 0.01), and down-regulating that of anti-apoptosis gene Bcl-2 (P < 0.05).</p><p><b>CONCLUSION</b>Local application of allicin via gastroscopy can inhibit the cell growth and proliferation of progressive gastric carcinoma, and can also promote gastric carcinoma cell apoptosis.</p>

Effects of allicin on changes of hemorheology in focal cerebral ischemia-reperfusion injury / 中国中药杂志

<p><b>OBJECTIVE</b>To explore the effects of allicin on the changes of hemorheology in focal cerebral ischemia-reperfusion injury in rats.</p><p><b>METHOD</b>Middle cerebral artery occlusion (MCAO) was used to make focal cerebral ischemia-reperfusion model by intravascular nylon filament occlusion. The protective effects of allicin at different doses were evaluated by investigating neurological function score, infarction volume and water content of brain. The changes of blood rheology were detected.</p><p><b>RESULT</b>Compared with model group, allicin (15, 25 mg x kg(-1)) increased the neurological function score and decreased the water content and infarction volume of brain in rats. Allicin (15, 25 mg x kg(-1)) inhibited the increasing of the blood viscosity, high shear rate reduced viscosity, high shear relative reduced viscosity and low shear relative reduced viscosity.</p><p><b>CONCLUSION</b>Allicin has protective effects on cerebral ischemia-reperfusion injuries. The mechanism may be related to inhibit the increasing of hemorheology.</p>

Protective role of allicin and L-ascorbic acid against the genotoxic damage induced by chlormadinone acetate in cultured human lymphocytes.

In our present study, different doses of allicin and L-ascorbic acid were tested against the genotoxic damage induced by chlormadinone acetate (CMA; 40 microM) using chromosomal aberrations (CAs) and sister chromatid exchanges (SCEs) as the parameters. Treatment with allicin and L-ascorbic acid resulted in reduction of CAs and SCEs. The results suggested a protective role of allicin and L-ascorbic acid against CMA induced genotoxic damage.

Antigenotoxic effect of allicin against methyl methanesulphonate induced genotoxic damage.

Allicin, one of the sulfur compounds especially thiosulphonates of garlic (Allium sativum), possesses antioxidant and thioldisulphide exchange activity and is also shown to cause a variety of actions potentially useful for human health. In this investigation we determined its antigenotoxic potential using chromosomal aberrations (CAs) and sister chromatid exchanges (SCEs) induced by methyl methanesulphonate (MMS) as genotoxic end points both in the presence as well as absence of rat liver microsomal activation system (S9 mix) in cultured human lymphocytes. We tested the effect of 5, 10 and 20 microM of allicin on the damage exerted by 60 microM of MMS. The levels of CAs and SCEs were lowered suggesting an antigenotoxic role of allicin against genotoxic damage both in the presence as well as absence of metabolic activation.

Pre- planting evaluation of allicin content and botanical traits in Iranian garlic [Allium sativum L.] ecotypes

Garlic is one of the medicinal plants, which is efficate in treatment of mild hypertension and lipid profiles. Along this study allicin content and botanical traits are evaluated in pre-planting stage. Bulb mean weight; clove mean weight and clove number per bulb were the evaluated morphological characters. Allicin content evaluation was done by HPLC. The correlation existed between evaluated characters were analyzed. We found that all samples were rich in allicin with values higher than pharmaceutical grade [4.5 mg/g]. This study showed that ecological conditions had not detectable relation with allicin content. A cluster analysis of data was performed based on morphological characters and allicin content. In general we did not detect significant relationship between genetic diversity and geographical origins, which suggests that probably the genetic factors have more influence than ecology

The effect of cell killing and apoptosis by human herpes simplex virus- thymidine kinase/ganciclovir system combined with allitride in BIU87 cells / 中华外科杂志

<p><b>OBJECTIVE</b>To study the killing effect of human herpes simplex virus-thymidine kinase/ganciclovir (HSV-TK/GCV) system combined with allitride and the possible apoptosis mechanism in BIU87 cells.</p><p><b>METHODS</b>The cytotoxicity after combination were estimated by theamine blue tetrazolium bromide (MTT). The morphological changes were observed with inverted microscope and in-situ cell apoptosis detection kit. Changes of apoptosis rate and cell cycle were assessed by flow cytometry. B-cell lymphoma-2 (bcl-2), bax, caspase-3 (cysteine aspartate specific proteinase) mRNA changes were detected by reverse transcriptase polymerase chain reaction, and caspase-3 activity was estimated with colorimetry.</p><p><b>RESULTS</b>For combination group, the cell killing rate was raised to 72.50% to compare with 35.00% of GCV and 37.00% of allitride separately and there was a synergistic effect between these two drugs. The cell apoptosis was induced in all three groups and for the combination group the time of S-phase and G(2)-phase arrest were earlier than other two groups. Both drugs could inhibit the expression of bcl-2 and promote the expression and activity of caspase-3.</p><p><b>CONCLUSIONS</b>The combination of HSV-TK/GCV system with allitride can inhibit the proliferation of BIU87 cells congenerously through apoptosis, which may be correlated with S- and G(2)-phase arrest, down-regulation of bcl-2 and increased caspase-3 expression and its activity.</p>

Antigenotoxic effect of allicin against SCEs induced by methyl methanesulphonate in cultured mammalian cells.

Allicin, one of the sulphur compounds of garlic (Allium sativum), possesses antioxidant and thiol disulphide exchange activity and is also shown to cause a variety of activities potentially useful for human health. In this investigation, the effect of 1,5,10 and 20 microM of allicin was determined for inhibiting the rate of SCE induced by 60 microM of MMS. Cultured human lymphocytes from two female donors were used for the experiment. The levels of SCEs were lowered by allicin suggesting its antigenotoxic activity in mammalian cells in vitro.