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Bioremediation of municipal sludge by vermitechnology and toxicity assessment by Allium cepa.

The aim of this study was to evaluate municipal sludge (MS) for its toxic potential by Allium cepa and also to understand the effect of vermicomposting on the reduction of toxicity, if any. Municipal sludge (MS) and vermicomposted sludge (VS) were evaluated. Elemental analysis of MS showed the presence of heavy metals. Morphological studies of A. cepa roots indicated coiled and wavy roots on exposure to MS but no root abnormality was reported in VS. Under genotoxic studies, inhibition in mitotic index was concentration dependent and the control values of 11.76 gradually reduced to 5.40 at 10% MS leachate whereas mitotic index was increased to 9.48 at 10% VS leachate. Exposure of leachate induced chromosomal aberrations, micronucleus formation and binucleate cells in a dose dependent manner. However, mitotic aberrations were observed significant at 10% MS leachate but they were insignificant at 10% VS leachate. The wet and dry weight of roots, root elongation and chlorophyll contents were reduced as the concentration of leachate increased but VS leachate did not produce considerable reduction. The wet and dry weight of A. cepa roots were 20.312 g and 3.250 g respectively and they were reduced to 10.82 g and 1.68 g respectively at 10% MS leachate but VS leachate showed an increase to 18.127 g and 2.53 g respectively. Total chlorophyll in control, 10% MS leachate and 10% VS leachate were 0.245 g, 0.162 g and 0.214 g respectively. It could be concluded that the MS was toxic to a remarkable extent but vermicomposting of sludge might be beneficial for bioremediation and recommended before land filling.

Animals↗

Development of a disposable pyruvate biosensor to determine pungency in onions (Allium cepa L.).

A disposable prototype pyruvate biosensor was constructed using pyruvate oxidase immobilised on mediated meldolas blue electrodes to determine pungency in onions (Allium cepa L.). The optimum operating potential was +150 mV (versus Ag/AgCl). A strong correlation between the biosensor response and untreated onion juice of known pyruvate concentration 2-12 micromol/g fresh weight (FW) was demonstrated. The biosensor was able to differentiate between low and high pungency onions. The detection limit using 1 unit of pyruvate oxidase was 1-2 micromol/g FW. Optimum concentrations of co-factors TPP, FAD and MgSO4 comprising the enzyme cocktail were determined as being 0.04, 0.1 and 30 mM, respectively.

Biosensing Techniques↗

Cadmium accumulation in Allium schoenoprasum L. grown in an aqueous medium.

The ability of Allium schoenoprasum L. (chives) to accumulate and tolerate cadmium in aqueous Hoagland medium at 50microM and 250microM was tested under continuous growth or several successive harvests of shoots. After 28 days of continuous growth, chives accumulated the metal up to 0.2% and 0.5% of its dry weight, when grown in 50microM and 250microM, respectively. In experiments that the leaves were successively harvested every 16 days, there were no obvious stress symptoms after six harvests during a period of 96 days at 50microM Cd. At 250microM, after 64 days and four harvests, inhibition of growth occurred. In each treatment, a total of 1.2g kg(-1) DW and 2.4g kg(-1) DW was accumulated in the leaves, respectively. Total SH compounds concentration in leaf was found significantly higher by 3 and 7.4 times in plants treated with Cd at 50microM and 250microM in comparison to the control, respectively, while no difference in the concentration of glutathione (GSH+GSSG) was found. Thus, it is assumed that sulphur-containing compounds, yet unknown, are involved in defensive mechanisms against heavy metals in chives. The results presented, point to chives phytoremediation potential, but also on the potential risk in accumulation of heavy metals in a commonly edible plant.

Analysis of Variance↗

Effect of red pepper Capsicum annuum var. conoides and garlic Allium sativum on plasma lipid levels and cecal microflora in mice fed beef tallow.

Antihyperlipidemia or hypocholesterolaemic and antibacterial activities of red hot pepper and garlic are well known. To determine the effect of the dietary spices ingested to suppress blood lipids on the intestinal condition, we examined plasma lipid levels and cecal microflora in mice that were fed diets containing 19% (w/w) beef tallow and 2% red pepper Capsicum annuum var. conoides 'Takanotume' (RP) or garlic Allium sativum 'White' (GP) for 4-weeks. Plasma triacylglyceride level was suppressed by the spices. RP lowered cecal bacteroidaceae, a predominant bacterial group (from 9.4 to 9.0 log CFU/g), bifidobacteria (from 8.7 to 7.6 log CFU/g), and staphylococci. Although GP increased the cecal weight including their contents, significant differences were not shown in the cecal microflora. These results suggest that RP can affect the intestinal condition and host health through the disturbance of intestinal microflora.

Animals↗

Haematological evaluation of ethanolic extract of Allium ascalonicum in male albino rats.

The haematological effect of ethanolic extract of Allium ascalonicum was evaluated in male albino rats during a 21 day administration at the doses of 50, 100 and 200 mg/kg b.w, orally. Parameters evaluated include the serum lipids, red and white cell indices. The results showed that the extract administered decreased most of the parameters relating to red cell and increased most of those parameters relating to white cells. It also decreased the total cholesterol (TCH), high density lipoprotein cholesterol (HDL) and low density lipoprotein cholesterol (LDL) with no significant effect on the triglyceride levels.

Administration, Oral↗

Enhancement of peritoneal macrophage phagocytic activity against Leishmania major by garlic (Allium sativum) treatment.

It has been shown that garlic (Allium sativum) extract modulates immune responses. Macrophage activation is required to establish control of the intracellular infection and progressive disease of leishmaniasis. In this study, we consider the effect of a garlic extract and a fraction isolated from it on the engulfment and destruction of Leishmania major by resident peritoneal macrophages of Balb/c mice. In regard to this study, the infiltration of macrophages in the peritoneal cavity after garlic treatment and the rate of amastigotes per macrophage were determined. The results show that a single dose of 20 mg/kg garlic extract intraperitoneally (i.p.) alters the number of peritoneal macrophages for at least 2 weeks. Intraperitoneal injection of garlic extract (20 mg/kg) or its protein fraction (0.04 mg/kg) augments parasite engulfment and destruction of intracellular amastigotes by macrophages.

Animals↗

Lipid lowering effect of S-methyl cysteine sulfoxide from Allium cepa Linn in high cholesterol diet fed rats.

The lipid lowering action of S-methyl cysteine sulfoxide (SMCS) isolated from Allium cepa Linn (family: Liliaceae) was investigated in Sprague-Dawley rats fed on 1% cholesterol diet, in comparison to the hypolipidemic drug gugulipid. Administration of SMCS at a dose of 200mg/kg body weight for 45 days ameliorated the hyperlipidemic condition. The lipid profile in serum and tissues showed that concentrations of cholesterol, triglyceride and phospholipids were significantly reduced when compared to their untreated counterparts. The total lipoprotein lipase activity in the adipose tissue was decreased with also a decrease in the free fatty acid levels in serum and tissues. The activities of the lipogenic enzymes glucose 6-phosphate dehydrogenase and malic enzyme as also of HMG CoA reductase in the tissues remained low on treatment indicating that both the drugs did not favor lipogenesis and cholesterogenesis in the hyperlipidemic animals. The fecal excretion of bile acids and sterols was further increased upon treatment with the drugs. The results are directive to that both gugulipid and SMCS cause reduction of endogenous lipogenesis, increase catabolism of lipids and subsequent excretion of metabolic by-products through the intestinal tract. However, gugulipid is a better drug than SMCS at a low dose of 50mg/kg body weight.

Animals↗

Cadmium accumulation and oxidative burst in garlic (Allium sativum).

To investigate the temporal sequence of physiological reactions of garlic (Allium sativum) to cadmium (Cd) treatment, seedlings developed from cloves were grown in increasing concentrations of CdCl2, ranging from 1-10 mM, for up to 8 days in sand. Analysis of Cd uptake indicated that most Cd accumulated in roots, but some was also translocated and accumulated in leaves at longer exposure time (after 12h) and higher concentrations (5 and 10mM) of CdCl2. Changes in activities of antioxidative enzymes, including superoxide dismutase (SOD), peroxidase (POD) and catalase (CAT), were characterized in leaves of garlic seedlings. Cd (5 and 10 mM) initially inhibited the activities of SOD and CAT but thereafter recovered or even increased compared with control plants. POD activities at 5 and 10 mM of Cd increased more than 3-4 times over control plants within 12 h and then dropped, but were still higher than controls at the end of the experiment. Otherwise lipid peroxidation enhanced with the increasing of incubation time and concentrations of external Cd. Leaves exposed to 1 mM CdCl2 showed a less pronounced response and only a small reduction in shoot growth. These results suggested that in leaves of garlic seedlings challenged by CdCl2 at higher concentrations, induction of these various enzymes is part of a general defense strategy to cope with overproduction of reactive oxygen. The possible mechanism of antioxidative enzymes changing before Cd accumulation in leaves of garlic seedlings is discussed.

Cadmium Chloride↗

Salicylic acid triggers genotoxic adaptation to methyl mercuric chloride and ethyl methane sulfonate, but not to maleic hydrazide in root meristem cells of Allium cepa L.

Salicylic acid (SA), 0.01 mM, a signalling phytohormone, was tested for induction of adaptive response against genotoxicity of methyl mercuric chloride (MMCl), 0.013 mM; ethylmethane sulfonate (EMS), 2.5 mM, or maleic hydrazide (MH), 5 mM, in root meristem cells of Allium cepa. Induction of adaptive response to EMS by hydrogen peroxide (H2O2), 1 mM, and yet another secondary signal molecule was tested for comparison. Assessed by the incidence of mitoses with spindle and/or chromosome aberration and micronucleus, the findings provided evidence that SA-conditioning triggered adaptive response against the genotoxic-challenges of MMCl and EMS, but failed to do so against MH. H2O2, which is known to induce adaptive response to MMCl and MH, failed to induce the same against EMS in the present study. The findings pointed to the possible role of signal transduction in the SA-induced adaptive response to genotoxic stress that perhaps ruled out an involvement of H2O2.

Adaptation, Biological↗

The influence of garlic (Allium sativum) extract on interleukin 1alpha-induced expression of endothelial intercellular adhesion molecule-1 and vascular cell adhesion molecule-1.

Inflammation plays an important role in both the initiation of atherosclerosis and development of atherothrombotic events. The adherence of leukocytes/monocytes to the endothelium is an early event in atherogenesis. Phytotherapeutica as garlic and garlic extracts were shown to have beneficial modulating effects in patients with atherosclerotic disease. The aim of this study was to evaluate in vitro the influence of water-soluble garlic (Allium sativum) extract on the cytokine-induced expression of endothelial leukocyte adhesion molecules such as intercellular adhesion molecule-1 (ICAM-1, CD54) and vascular cell adhesion molecule-1 (VCAM-1, CD106). Cytokine-induced expression of cellular adhesion molecules was measured on primary human coronary artery endothelial cell (HCAEC) cultures. HCAEC were cultured in microvascular endothelial cell growth medium and preincubated with garlic extract at various concentrations (0.25-4.0 mg/ml), after which human interleukin-1alpha (IL-1alpha, 10 ng/ml) was added for 1 day. Fluorescein isothiocyanate (FITC)-labeled anti-ICAM-1 and FITC-labeled anti-VCAM-1 were used to analyze the IL-1alpha-induced expression of ICAM-1 and VCAM-1 by flow cytometry. Incubation of HCAEC with garlic extract significantly decreased ICAM-1 and VCAM-1 expression induced by IL-1alpha. In addition, we examined the effects of garlic extract on the adhesion of monocytes to endothelial cells, using the monocytic U937 cell line. The presence of garlic extract significantly inhibited the adhesion of monocytes to IL-1alpha-stimulated endothelial cells. These results indicate that garlic extract modulates the expression of ICAM-1 and VCAM-1, thus potentially contributing to the beneficial effects traditionally attributed to garlic.

Cell Adhesion↗

Antidiabetic effect of garlic (Allium sativum L.) in normal and streptozotocin-induced diabetic rats.

OBJECTIVE: The antidiabetic effect of garlic ethanolic extract (Allium sativum L.) was investigated in normal and streptozotocin-induced diabetic rats. RESEARCH METHODS AND PROCEDURE: In the present study, oral administration of garlic extract (0.1, 0.25 and 0.5 g/kg body wt.) for 14 days on the level of serum glucose, total cholesterol, triglycerides, urea, uric acid, creatinine, aspartate amino transferase (AST) and alanine amino transferase (ALT) in normal and streptozotocin-induced diabetic rats were evaluated. RESULTS: Oral administrations of the garlic extract significantly decreased serum glucose, total cholesterol, triglycerides, urea, uric acid, creatinine, AST and ALT levels, while increased serum insulin in diabetic rats but not in normal rats (p<0.05). A comparison was made between the action of garlic extract and glibenclamide (600 microg/kg), the known antidiabetic drug. The antidiabetic effect of the extract was more effective than that observed with glibenclamide. CONCLUSION: It is concluded that the plant must be considered as excellent candidate for future studies on diabetes mellitus.

Alanine Transaminase↗

Anthocyanins with 4'-glucosidation from red onion, Allium cepa.

The anthocyanins, cyanidin 3-O-(3"-O-beta-glucopyranosyl-6"-O-malonyl-beta-glucopyranoside)-4'-O-beta-glucopyranoside, cyanidin 7-O-(3"-O-beta-glucopyranosyl-6"-O-malonyl-beta-glucopyranoside)-4'-O-beta-glucopyranoside, cyanidin 3,4'-di-O-beta-glucopyranoside, cyanidin 4'-O-beta-glucoside, peonidin 3-O-(6"-O-malonyl-beta-glucopyranoside)-5-O-beta-glucopyranoside and peonidin 3-O-(6"-O-malonyl-beta-glucopyranoside) have been isolated in minor amounts from pigmented scales of red onion, Allium cepa, in addition to six known anthocyanins. The structures were established mainly by extensive use of 2D NMR spectroscopy and electrospray LC-MS. With exception of cyanidin 4'-glucoside and cyanidin 3,4'-diglucoside reported from Hibiscus esculentus with inadequate documentation, this is the first identification of anthocyanins with 4'-glycosidation. Compared to cyanidin 3-glycosides the cyanidin 4'-glucoside derivatives showed hypsochromic shifts of visible lambda(max) and hyperchromic effects on wavelengths around 440 nm, similar to pelargonidin 3-glycosides.

Anthocyanins↗

Certain antioxidant enzymes of Allium cepa as biomarkers for the detection of toxic heavy metals in wastewater.

The utility of antioxidant enzymes, viz glutathione-S-transferase (GST), glutathione reductase (GR), superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX), glutathione peroxidase (GPX), monodehydroascorbate reductase (MDHAR) and dehydroascorbate reductase (DHAR), as biomarkers of heavy metal pollution in water was investigated using the Allium cepa (onion) system. These antioxidant enzymes were assayed in onion bulbs exposed to certain heavy metals taken separately, the test metals taken in combination as well as the industrial wastewater especially found to contain these metals. GST exhibited significantly enhanced activity upon treatment with individual heavy metals. However, GR, SOD and CAT did not show such a pronounced increase in activities. At higher heavy metal concentrations, GR, SOD and CAT showed a steep decline while GST activity still showed a rise. Moreover, APX, GPX and MDHAR also exhibited remarkable induction with increase in the concentration of individual heavy metals. However, there was no significant change in DHAR activity with respect to the controls. Metabolites like ascorbate (ASC) and glutathione (GSH) exhibited significant decline with increase in the concentration of individual heavy metals while the level of H(2)O(2) continued to display the rise up to a heavy metal concentration of 100 microM, after which it showed a gradual decline. A. cepa bulbs treated with wastewater sample showed enzyme activity profiles similar to that shown with heavy metals, thereby suggesting the presence of heavy metals in the test wastewater. Atomic absorption spectrophotometry also detected large amounts of Cd, Cr, Cu, Hg, Pb and Zn in the test water sample. The metal mixture, containing the amounts of heavy metals equivalent to those found in the wastewater, resulted in steep declines in GR, SOD and CAT activities in A. cepa while GST showed a rise. However, when this metal mixture was diluted to 2000-fold, GR, SOD and CAT also showed enhanced activities compared with the controls. Contrary to the above finding, APX, GPX and MDHAR exhibited the rise in activities in A. cepa exposed to the metal mixture at all dilutions. In the presence of cycloheximide, all the enzymes returned to the levels of untreated controls while chloramphenicol did not have any effect on the test enzymes, thereby suggesting de novo protein synthesis of the test antioxidant enzymes in the cytosolic compartment of the cell as a result of exposure to the heavy metals.

Antioxidants↗

Comparative biomonitoring of leachates from hazardous solid waste of two industries using Allium test.

Hazardous industrial wastes are inevitable source of environmental pollution. Leachates from these wastes might contaminate the origins of potable water and affect human health. The study was carried out to determine the possible genotoxic effects of leachates from solid waste of a metal and dye industry using the Allium cepa chromosome aberrations assay. The 10% leachates were prepared from solid wastes obtained from both the industries and examined for the presence of heavy metal content and genotoxicity. To simulate the field and laboratory conditions, A. cepa bulbs were exposed through soil and aqueous medium for 48 h to 2.5-10% leachates. The results revealed that both metal waste leachate (MWL) and dye waste leachate (DWL) contained high concentrations of chromium, nickel and iron that significantly induced cytogenetic alterations. Significant inhibition of mitotic index (MI), inductions of chromosomal/mitotic aberrations (CA/MA) and micronuclei (MN) formation were found in all experimental groups exposed to MWL and DWL. The effects observed were concentration dependent and the frequency of aberrations was higher with treatment of MWL than DWL. The MI was severely inhibited at 10% aqueous exposure it was 4.59+/-0.69 (P<0.001) in MWL and almost half to that induced by DWL that was 8.62+/-0.69 (P<0.05). Significant frequency of CA/MA and MN induced by MWL was 14.21 (P<0.001) and 0.33 (P<0.001) whereas CA/MA and MN induced by DWL was 7.81 (P<0.001) and 0.13 (P<0.05) in the aqueous medium. The investigations inferred that abnormalities caused by MWL were higher than DWL both in soil and aqueous media. These toxic responses may have relied on raised heavy metal concentrations of metal-based than dye industrial wastes.

Chromosome Aberrations↗

Comparison of distillation and ultrasound-assisted extraction methods for the isolation of sensitive aroma compounds from garlic (Allium sativum).

A comparative study of traditional simultaneous distillation extraction (SDE), microwave assisted hydrodistillation extraction (MWHD) and ultrasound-assisted extraction (USE) is presented, for the extraction of essential oils from fresh garlic (Allium sativum) cloves. Each method is evaluated in terms of qualitative and quantitative composition of the isolated essential oil. The highly reactive sulfur molecules of the garlic volatile fraction show variable response to the different isolation methods. The application of ultrasound for the extraction of the essential oil is considered to cause a lesser damage of thermal-sensitive molecules, thus, providing a better approach of the compounds primarily responsible for the characteristic odor and taste of freshly chopped garlic. All heat-involving isolation procedures have been shown to differentiate the volatile-fraction profile as analyzed by GC-MS. Especially when grouping the compounds into cyclic and acyclic, the percentage concentrations drop from 77.4% to 8.7% for the acyclic while that of the cyclic compounds increase from 4.7% to 70.8%. The observed fact may be attributed to the effect of the heat applied, which changes from harsh thermal treatment (SDE) to short time thermal (MWHD) and room-temperature isolation (USE). The use of USE proves to be crucial in order to provide reliable insight into garlic's chemistry.

Chemical Fractionation↗

Effects of garlic (Allium sativum L.) supplementation on cytochrome P450 2D6 and 3A4 activity in healthy volunteers.

Garlic (Allium sativum L.) is a commonly used food and herbal supplement. The objective of this study was to assess in healthy volunteers (N = 14) the influence of a garlic extract on the activity of cytochrome P450 (CYP) 2D6 and 3A4. Probe substrates dextromethorphan (CYP2D6) and alprazolam (CYP3A4) were administered orally at baseline and again after treatment with garlic extract (3 x 600 mg twice daily) for 14 days. Urinary dextromethorphan/dextrorphan ratios and alprazolam plasma concentrations were determined by HPLC at baseline and after garlic extract treatment. The ratio of dextromethorphan to its metabolite was 0.044 +/- 0.48 at baseline and 0.052 +/- 0.095 after garlic supplementation. There were no significant differences between the baseline and garlic phases (P > or =.05). For alprazolam, there were no significant differences in pharmacokinetic parameters at baseline and after garlic extract treatment (all P values > or =.05; maximum concentration in plasma, 27.3 +/- 2.6 ng/mL versus 27.3 +/- 4.8 ng/mL; time to reach maximum concentration in plasma, 1.9 +/- 1.4 h versus 2.4 +/- 1.8 h; area under the time-versus-concentration curve, 537 +/- 94 h. ng. mL(-1) versus 548 +/- 159 h. ng. mL(-1); half-life of elimination, 13.7 +/- 4.4 h versus 14.5 +/- 4.3 h). Our results indicate that garlic extracts are unlikely to alter the disposition of coadministered medications primarily dependent on the CYP2D6 or CYP3A4 pathway of metabolism.

Adult↗

Superiority of intravesical immunotherapy with Corynebacterium parvum and Allium sativum in control of murine bladder cancer.

Intravesical immunotherapy with bacillus Calmette-Guerin (BCG), Corynebacterium parvum (CP), keyhole limpet hemocyanin (KLH), and an extract of Allium sativum (AS) was studied in mice transplanted intravesically with mouse bladder tumor cells (MBT-2). Female C3H/He mice were anesthetized with sodium pentobarbital. Two X 10(5) MBT-2 cells were delivered into the bladder transurethrally using a small catheter, immediately after the posterior wall of the bladder had been electrocauterized. Bladder tumor became palpable or demonstrable microscopically in two weeks. Immunotherapy with BCG (2 X 10(6) CFU), CP (250 micrograms), KLH (50 micrograms), or AS (25 mg) was administered directly into the bladder via urethral catheter on day 1, day 6, or days 1 and 6. On day 21 the bladders and spleens were excised and weighed, and the bladders were examined macroscopically and microscopically for evidence of tumor. The results of the study showed that two treatments given one and six days after tumor transplant yielded the lowest tumor incidence and that CP and AS appeared equally effective or even slightly more effective than BCG in this model. These results suggest that clinical evaluation of CP or AS may be worthwhile.

Adjuvants, Immunologic↗

Superiority of intralesional immunotherapy with Corynebacterium parvum and Allium sativum in control of murine transitional cell carcinoma.

Immunotherapy with bacillus Calmette-Guerin (BCG), Corynebacterium parvum (CP), keyhole limpet hemocyanin (KLH) and an extract of Allium sativum (AS) was studied in a transitional cell carcinoma (MBT-2) in mice. Comparison was made between intraperitoneal (IP) versus intralesional (IL) administration of these agents. C3H/He mice were transplanted subcutaneously in the hind limb with 5 X 10(4) tumor cells. After transplantation, mice were randomized into groups to receive either IP or IL treatments with BCG (2 X 10(6) CFU), CP (250 micrograms.), KLH (50 micrograms.) or AS (25 mg.). At weekly intervals the tumor volume was determined. To assess the local cellular events following these treatments, histopathological studies were performed 10 days after tumor transplant with tissues removed from the injected sites. IL route was much more effective than IP route in inhibiting tumor growth. CP and AS exhibited more significant therapeutic effect than BCG or KLH. No tumor developed in mice which received five IL treatments of CP or AS. The data indicate that CP or AS may serve as effective biological response modifiers in controlling transitional cell carcinoma. The study further emphasizes that route and frequency of administration are crucial variables determining efficacy of immunotherapy.

Animals↗