Stoned on spices: a mini-review of three commonly abused household spices.

INTRODUCTION: Although spices are widely used as food products and are generally regarded as safe, intentional abuse of household spices may occur and is likely underreported in the medical literature. Spices are inexpensive and widely available for purchase by individuals of all ages and may be perceived as being safer than traditional drugs of abuse. DISCUSSION: Nutmeg, cinnamon, and vanilla are commonly abused spices. The major component of nutmeg is myristicin; myristicin has activity at serotonergic receptors and may result in psychomimetic symptoms after exposure. Cinnamon oils contain local irritants which may cause dermatitis or ulcerations after topical application. Ground cinnamon contains cellulose fibers; these are biopersistent and bioresistant, and inhalational exposure to cinnamon powder can result in chronic pulmonary inflammation and fibrosis. Pure vanilla extract contains a minimum of 35% ethanol according to the United States Food and Drug Administration standards, and abuse of vanilla extract may occur among individuals seeking ethanol intoxication. CONCLUSIONS: Overall, misuse or abuse of these spices frequently results in mild to moderate symptoms that do not require medical intervention, although more serious intoxications may require hospitalization. Clinicians should be aware of the potential dangers of household spice abuse and understand management strategies for these exposures.

A Review on Mycotoxins and Microfungi in Spices in the Light of the Last Five Years.

Spices are imported worldwide mainly from developing countries with tropical and/or subtropical climate. Local conditions, such as high temperature, heavy rainfall, and humidity, promote fungal growth leading to increased occurrence of mycotoxins in spices. Moreover, the lack of good agricultural practice (GAP), good manufacturing practice (GMP), and good hygienic practice (GHP) in developing countries are of great concern. This review summarizes recent data from a total of 56 original papers dealing with mycotoxins and microfungi in various spices in the last five years. A total of 38 kinds of spices, 17 mycotoxins, and 14 microfungi are discussed in the review. Worldwide, spices are rather overlooked in terms of mycotoxin regulations, which usually only cover aflatoxins (AFs) and ochratoxin A (OTA). In this paper, an extensive attention is devoted to the limits on mycotoxins in spices in the context of the European Union (EU) as well as other countries. As proven in this review, the incidence of AFs and OTA, as well as other mycotoxins, is relatively high in many spices; thus, the preparation of new regulation limits is advisable.

A botanical from the antiproliferative Cameroonian spice, Imperata cylindrica is safe at lower doses, as demonstrated by oral acute and sub-chronic toxicity screenings.

BACKGROUND: The cytotoxicity of the root's methanol extract of Imperata cylindrica (ICR). was previously reported in a panel of human cancer cell lines, including multi-drug resistant phenotypes. The aim of this study was to assess the acute and sub-chronic oral toxicity of methanol root extract of Imperata cylindrica. METHODS: The acute toxicity was carried out according to the experimental protocol of OECD. The plant extract was administered orally to female rats at a single dose of 5000 mg/kg for 14 days and the animals were observed for any behavioral changes or mortality. For sub-chronic toxicity study, ICR was orally administered daily to male and female rats at different doses (250, 500 and 1000 mg/kg per b.w.) for 30 days. During these treatment days the animals were observed for any appearance of toxicity symptoms; following the treatment period, animals were sacrificed for hematological, biochemical and histopathology analysis. RESULTS: From the results of the acute oral toxicity assay, ICR was found to be non-toxic at the dose of 5000 mg/kg b.w. During the period of sub-chronic toxicity test, observation of signs, behavior and health status of the animals showed no abnormality in the groups of animals treated with ICR as compared to the controls. Significant variation of the relative body weights of heart and kidney were observed at dose a 1000 mg/kg b.w. Significant decrease of aspartate aminotransferase, creatinine level, low density lipoprotein concentration, triglyceride and total cholesterol were observed. In males, we noticed a significant decrease of the level of granulocytes with an increase of lymphocytes and mean corpuscular hemoglobin concentration levels. Histological examinations performed on kidney and liver showed a normal kidney architecture and liver also presented a normal hepatic architecture with slight degeneration at a dose 1000 mg/kg b.w. CONCLUSION: ICR is safe for acute oral administration; however, for long-term oral administration, safety measures should be taken. Thus, oral sub-chronic exposure of ICR at lower doses are recommended while higher doses around 1000 mg/kg b.w. should be discouraged.

Quality variation and standardization of black pepper (Piper nigrum): A comparative geographical evaluation based on instrumental and metabolomics analysis.

Black pepper (Piper nigrum; BP), known as the 'king of spices', imported from various countries is widely available in Saudi Arabian markets, as its demand as a food as well as a medicine for minor ailments is increasing in the country. However, there is a lack of appropriate information regarding these samples in terms of quality variation and standardization. We thus aimed to evaluate the quality and standardize the BP sample with respect to its physicochemical characters, active principle variation [i.e. piperine (PPN)], toxicity, and biological activity. The main focus is to validate whether any difference exists in the quality and quantity of active principle in these samples. For this purpose, physicochemical (chemical tests and ash values) and instrumental analyses [accelerated solvent extraction (ASE), ultra-high-pressure liquid chromatography (UHPLC)-diode array detector, infrared (IR), nuclear magnetic resonance (NMR), and inductively coupled plasma-MS (ICP-MS)] and biological evaluation {in vitro antioxidant activity [2,2-diphenyl-1-picrylhydrazyl and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)] and cytotoxicity assay} were performed. An extract yield (g) with %recovery of 2.26 ± 4.24 (11.3) was obtained for the Vietnamese sample, using a fast and rapid method of extraction (ASE). These values were 1.22 ± 2.64 (6.1) and 0.75 ± 1.69 (3.75) for the Pakistani and Indian samples. Physicochemical tests revealed the presence of flavonoids and phenolic compounds in all samples; however, in the Vietnamese sample a low amount of total, acid-insoluble, and high water-soluble ash value was noted. IR and NMR was applied to further standardize the samples. Results of ICP-MS analysis showed a high amount of macronutrients and micronutrients in the samples tested while UHPLC analysis revealed a high amount of PPN (ng/mL) in the Pakistani sample (1,362,614.09); these values were 1,051,848.04 and 768,512.81 for the Vietnamese and Indian samples, respectively. In vitro antioxidant and cytotoxicity activities revealed higher potential for the Vietnamese sample. The samples were properly standardized and effectively differentiated in terms of quality and biological activities using a fast and reliable method, however it certainly does not mean that a particular geographical region is more better or productive in terms of herbal products.

Assessment of Occupational Health Hazards Due to Particulate Matter Originated from Spices.

Spices have been known for their various health activities; however, they also possess the allergic potential for the respiratory system and the skin as they are fine particulate matter. Persons involved in spice agriculture and food industries are at greater risk since they are exposed to a considerable amount of combustible dust, which may be the cause of fire and explosion and adversely affect the health. These workers may experience allergy, long-term and short-term respiratory issues including occupational asthma, dermatitis, etc. Some spices induce T cell-based inflammatory reaction upon contact recognition of the antigen. Antigen Presenting Cells (APC) on binding to the causative metabolite results in activation of macrophages by allergen cytokine interleukin (IL)-12 and tumor necrosis factor-beta (TNF). Cross-reactivity for protein allergens is another factor which seems to be a significant trigger for the stimulation of allergic reactions. Thus, it was imperative to perform a systematic review along with bioinformatics based representation of some evident allergens has been done to identify the overall conservation of epitopes. In the present manuscript, we have covered a multifold approach, i.e., to categorize the spice particles based on a clear understanding about nature, origin, mechanisms; to assess metabolic reactions of the particles after exposure as well as knowledge on the conditions of exposure along with associated potential health effects. Another aim of this study is to provide some suggestions to prevent and to control the exposure up to some extent.

In vitro toxicity evaluation of estragole-containing preparations derived from Foeniculum vulgare Mill. (fennel) on HepG2 cells.

Estragole, a common component of herbs and spices, is a wellknown genotoxic hepatocarcinogen in rodents, whereas its potential toxic effect in humans is still debated. In the European contest, one of the major sources of human exposure to this phytochemical is Foeniculum vulgare Mill. (fennel). Therefore, the aim of this study was to evaluate the in vitro toxicity of estragole in the context of two complex phytochemical mixtures derived from fennel: fennel seed powder (FSPw) and fennel seed essential oil (FSEO). The estragole-containing preparations were analysed for their ability to cause cytotoxicity, genotoxicity, apoptosis and cell cycle perturbation in the human hepatoma (HepG2) cell line. None of the tested concentrations of FSPw induced DNA damage, nor apoptosis or cell cycle perturbation. Although FSEO did not induce any genetic damage as well, it exerted marked dose-dependent apoptotic effects on HepG2 cells with a concurrent cell cycle arrest in G2/M at the highest tested dose. Although prospective analyses are required to clarify the observed toxic effects of FSEO, our results support the hypothesis that the genotoxicity of estragole may be significantly reduced or null in the context of botanical mixtures.

Nutmeg poisonings: a retrospective review of 10 years experience from the Illinois Poison Center, 2001-2011.

Nutmeg is a commonly consumed spice. The toxic effects of nutmeg have been purported to be due mainly to myristicin oil. Prior poison center series of nutmeg exposures show very few unintentional exposures of nutmeg to children younger than 13. Case series from these centers did not record drug exposures combined with nutmeg. This study is a review of Illinois Poison Center (IPC) data regarding nutmeg exposures from January of 2001 to December 2011. The goal of this study was to compare the Illinois data to the literature as well as look for current trends in nutmeg poisonings. The data were extracted using the code for hallucinogenic plants in the IPC database, and poisonings unrelated to nutmeg exposure were eliminated. Medical outcomes were noted as recorded. Thirty-two cases of nutmeg ingestion were reported. Of the 17 (53.1 %) unintentional exposures, 10 subjects (58.8 %) were under the age of 13. Four of the exposures in children under the age of 13 were ocular exposures. Fifteen exposures (46.9 %) were intentional exposures. Of these intentional exposures, five (33.3 %) were recorded to have combined drug intoxication. All of these were between the ages of 15 and 20. One patient with polypharmaceutical exposure required ventilatory support in the hospital. Our study shows an unexpected percentage of unintentional exposures in juveniles under the age of 13, out of the total exposures to nutmeg. Mixing of nutmeg with other drugs was seen and required more intervention in adolescents. More education about these two factors, i.e., nutmeg exposures as intentional polypharmacy in adolescents and unintentional exposures in young children, is advised.

[Advances in research of chemical constituents and pharmacological activities of common used spices].

Spices have enjoyed a long history and a worldwide application. Of particular interest is the pharmaceutical value of spices in addition to its basic seasoning function in cooking. Concretely, equipped with complex chemical compositions, spices are of significant importance in pharmacologic actions, like antioxidant, antibacterial, antitumor, as well as therapeutical effects in gastrointestinal disorders and cardiovascular disease. Although increasing evidences in support of its distinct role in the medical field has recently reported, little information is available for substantive, thorough and sophisticated researches on its chemical constituents and pharmacological activities, especially mechanism of these actions. Therefore, in popular wave of studies directed at a single spice, this review presents systematic studies on the chemical constituents and pharmacological activities associated with common used spices, together with current typical individual studies on functional mechanism, in order to pave the way for the exploitation and development of new medicines derived from the chemical compounds of spice (such as, piperine, curcumin, geniposide, cinnamaldehyde, cinnamic acid, linalool, estragole, perillaldehyde, syringic acid, crocin).

Protective effect of Eleteria cardamomum (L.) Maton against Pan masala induced damage in lung of male Swiss mice.

OBJECTIVE: To study the potential ameliorating properties of cardamom Elettaria cardamomum (E. cardamomum) L. Maton against pan masala induced damage in lung of male Swiss mice. METHODS: The experimental animals were divided into 3 groups (control, pan masala treated group and pan masala with cardamom treated group) to evaluate pan masala toxicity. The observations were substantiated with profound changes in the lung tissue as revealed in the histologic and transmission electron microscopic examinations. RESULTS: Lung of pan masala treated group showed adenocarcinoma, edema, and inflammation with increased activity of acid phosphatase, alkaline phosphatase, and lactate dehydrogenase. The deleterious effects were seen to be less in cardamom treated group and the enzymatic activity also decreased significantly (P<0.05) in the ameliorating group. CONCLUSIONS: Thus, the present experiment exciting results are observed when cardamom is supplemented with pan masala, or when given alone.

Toxicity evaluation of some traditional African spices on breast cancer cells and isolated rat hepatic mitochondria.

BACKGROUND: Fagara leprieuri (FL), Fagara xanthoxyloïdes (FX), Mondia whitei (MW) and Xylopia aethiopica (XA) are used in many African countries as food spices or in traditional medicine to treat several maladies. In this work, we (a) investigate whether the crude spice extracts present selective cytotoxicity for breast cancer cell lines and (b) investigate whether the same extracts affect the bioenergetics and calcium susceptibility of isolated liver mitochondrial fractions. RESULTS: All extracts were cytotoxic to the cell lines studied, with the exception of MW, which was less toxic for a normal cell line. Interestingly, some of the extracts did not depolarize mitochondria in intact breast cancer MCF-7 cells, although this effect was observed in a normal breast cancer cell line (MCF-12A). All extracts increased hepatic mitochondrial state 2/4 respiration and decreased the respiratory control ratio and the transmembrane electric potential. Also, the extracts induced the mitochondrial permeability transition (MPT). CONCLUSIONS: Mitochondrial toxicity may be part of the mechanism by which the spices tested cause inhibition of proliferation and death in the cell lines tested. This study also warrants caution in the excessive use of these spices for human consumption.

Spices, herbal xenobiotics and the stomach: friends or foes?

Spices and herbal remedies have been used since ancient times to treat a variety of disorders. It has been experimentally demonstrated that spices, herbs, and their extracts possess antimicrobial, anti-inflammatory, antirheumatic, lipid-lowering, hepatoprotective, nephroprotective, antimutagenic and anticancer activities, besides their gastroprotective and anti-ulcer activities. Despite a number of reports on the toxicity of herbs and spices, they are generally accepted as safer alternatives to conventional therapy against gastric ulcers. To this end, it is also believed, that excessive consumption of spices may favor the pathogenesis of gastric and duodenal ulcer and some studies have substantiated this common perception. Based on various in vivo experiments and clinical studies, on the effects of spices and herbs on gastric ulcers, it has indeed been shown that certain spices do possess remarkable anti-ulcer properties mediated by antisecretory, cytoprotective, antioxidant, and anti-Helicobacter pylori effects and mechanisms regulated by nitric oxide, prostaglandins, non-protein sulfhydryl molecules and epidermal growth factor expression. Accordingly, their consumption may attenuate and help prevent peptic ulcer disease. In the present review, the beneficial effects of spices and herbal nutritive components on the gastric mucosa are discussed against the paradigm of their deleterious potential.

Effect of alligator pepper (Zingiberaceae aframomum melegueta) on first trimester pregnancy in Sprague Dawley rats.

Alligator pepper (Aframomum melegueta) is a spice that is widely used in many cultures for entertainment, religious rites, food flavor and as a part of many traditional doctors medications. Pregnant women are among those who ingest Alligator pepper in these activities. This experiment was carried out to determine the health risk or benefit of Alligator pepper to pregnant women if any. Fifteen male rats and fifteen female rats of proven fertility from a pilot study were randomly paired in fifteen cages in a well ventilated room. After three days of mating, the males were withdrawn from the females, which were allowed to stay in their separate maternity cages for 18-25 days. The females in the control group were fed with normal rat chow and clean drinking water ad libitum for the duration of the experiment. Each of the rats in the experimental group was served 20 g of rat chow mixed with 50mg of Alligator pepper for one day only and thereafter fed with normal rat chow and clean drinking water ad libitum for 18-25 days. The rats in the control group had a mean of 7 litters each, while the rats in the experimental group did not litter at all. It was concluded that ingestion of large quantities of Alligator pepper poses a health risk to women in their first trimester of pregnancy.

Effect of aqueous extract of alligator pepper (Zingiberaceae aframomum melegueta) on gestational weight gain.

Increased gestational weight gain is associated with higher rates of complications of pregnancy and delivery. Gestational weight gain of 9-12 Kg has been associated with the best outcome for both mothers and infants. However, weight gain in most pregnant women is not within this range, perhaps due to the difficulty of calculating the exact quantity, timing and duration of dietary restriction in individual patients that would bring their weight gain within the normal range. There is therefore a need to develop a drug or food supplement that would reduce weight gain without causing adverse effects on the fetus. Aframomum melegueta is widely used in Nigeria by most people including pregnant women for various purposes. It is against this background that the present investigation examines the possibility of its beneficial effects on pregnancy, using Sprague Dawley rat as the animal model. Twenty female and ten male Sprague-Dawley rats of proven fertility from a pilot study were randomly mated in groups of two females and one male. Three days later, female rats in the experimental groups were given intra-peritoneal injections of 0.5 mg, 1 mg, 1.5 mg and 2 mg of aqueous extract of alligator pepper respectively while the control had 2 ml of distilled water. All rats were observed for 18-25 days. There was a significant [P<0.05] reduction in gestational weight gain of the experimental rats. The litters were not adversely affected. It is suggested that the active component of aqueous extract of alligator pepper be determined because of its beneficial effect of gestational weight gain reduction.

Anogenital allergic contact dermatitis, the role of spices and flavour allergy.

BACKGROUND: In patients with vulval or anogenital dermatitis, irritant contact dermatitis is more common than allergic contact dermatitis. The reported frequency and relevance of contact sensitivity in anogenital dermatitis varies greatly. OBJECTIVE: To determine the frequency and relevance of contact sensitization in a Dutch group of female patients with chronic anogenital complaints. METHODS: We reviewed patch test results of 53 women with chronic anogenital complaints, with sole vulval symptoms in 29 women and sole perianal in 5, in whom inflammatory skin diseases like lichen sclerosus, lichen planus, psoriasis, as well as infectious diseases were unlikely or excluded as a cause of their symptoms. All women were tested with the European baseline series plus additional test series according to their personal history. RESULTS: Thirty-five patients (66%) showed one or more positive test reactions. Seven of these patients (20%) had one or more clinically relevant positive reactions, most often to flavours and spices. CONCLUSION: A considerable number of patients with anogenital dermatitis have a contact sensitization. Clinically relevant reactions were mainly found to spices and flavours. This is in contrast to the data reported in the literature that shows most contact allergies in vulval patients to ingredients of topical medication.

Fumigant toxicity of essential oils from some common spices against pulse beetle, Callosobruchus chinensis (Coleoptera: Bruchidae).

In the present study, the essential oil from seven common spices, Anethum graveolens, Cuminum cyminum, Illicium verum, Myristica fragrans, Nigella sativa, Piper nigrum and Trachyspermum ammi was isolated and its insecticidal, oviposition, egg hatching and developmental inhibitory activities were determined against pulse beetle, Callosobruchus chinensis. Essential oils were isolated by hydrodistillation method using Clevenger apparatus. These essential oils caused death of adults and larvae of Callosobruchus chinensis when fumigated. The 24-h LC(50) values against the adults of the insect were 8.9 mul, 10.8 mul, 11.0 mul, 12.5 mul, 13.6 mul, 14.8 mul and 15.6 mul for N. sativa, A. graveolens, C. cyminum, I. verum, P. nigrum, M. fragrans and T. ammi oils respectively. On the other hand, against larval stage these values were 6.4 mul, 7.9 mul, 8.9 mul, 11.1 mul, 11.7 mul, 12.2 mul and 13.5 mul for N. sativa, A. graveolens, C. cyminum, I. verum, P. nigrum, M. fragrans and T. ammi respectively. These essential oils reduced the oviposition potential, egg hatching rate, pupal formation and emergence of adults of F(1) progeny of the insect when fumigated with sublethal concentrations. These essential oils also caused chronic toxicity as the fumigated insects caused less damage to the stored grains. The essential oil of N. sativa was found most effective against all the different stages of the Callosobruchus chinensis followed by A. graveolens, C. cyminum, I. verum, P. nigrum, M. fragrans and T. ammi oils. All the responses were found concentration-dependent. The toxic and developmental inhibitory effects may be due to suffocation and inhibition of various biosynthetic processes of the insects at different developmental stages.

Stress, diet and alcohol-induced oxidative gastrointestinal mucosal injury in rats and protection by bismuth subsalicylate.

Oxygen free radicals are implicated in the pathogenesis of stress and food/alcohol-induced gastrointestinal injury. We have investigated the effects of restraint stress, spicy food diet, high-fat diet and 40% ethanol on the enhanced production of reactive oxygen species, including superoxide anion and hydroxyl radicals, and on DNA fragmentation, lipid peroxidation and membrane microviscosity (indices of oxidative tissue damage) in gastric and intestinal mucosa of Sprague-Dawley rats. Furthermore, the protective ability of bismuth subsalicylate (BSS; 15 mg kg(-1) was determined against the gastrointestinal mucosal injury induced by these stressors. Animals on the high-fat diet consumed 31% more food as compared to other animals. Animals on the spicy food diet consumed ca. 23% more water as compared to control animals, and the high-fat diet animals consumed 17% less water. Restraint stress provided greater injury to both gastric and intestinal mucosa as compared to other stressors. Restraint stress, spicy food diet, high-fat diet and ethanol increased superoxide anion production by 10.0-, 4.3-, 5.7- and 4.8-fold, respectively, in the gastric mucosa, and by 10.4-, 5.3-, 7.0- and 5.5-fold in the intestinal mucosa. Exposure to restraint stress, spicy food diet, high-fat diet and 40% ethanol also increased hydroxyl radical production by ca. 14.3-, 4.5-, 3.5- and 4.8-fold, respectively, in the gastric mucosa, and by 17.0-, 4.8-, 3.5- and 4.7-fold in the intestinal mucosa. Bismuth subsalicylate administration to the animals provided significant protection against superoxide anion and hydroxyl radical production. Restraint stress, spicy food diet, high-fat diet and ethanol increased lipid peroxidation by 3.6-, 2.4-, 2.6- and 2.0-fold, respectively, in the gastric mucosa, and by 4.1-, 3.5-, 3.6- and 2.7-fold in intestinal mucosa. Administration of BSS decreased restraint stress, spicy food diet, high-fat diet and ethanol-induced gastric mucosal lipid peroxidation by ca. 26%, 36%, 45% and 18%, and intestinal mucosa lipid peroxidation by 20%, 21%, 46% and 42%, respectively. Approximately 4.0-, 2.0-, 2.4- and 2.0-fold increases in DNA fragmentation were observed in the gastric mucosa of rats exposed to restraint stress, spicy food diet, high-fat diet and 40% ethanol, respectively, and similar increases in the intestinal mucosa. These same four stressors increased membrane microviscosity by 11.6-, 6.1-, 7.3- and 5.4-fold, respectively, in the gastric mucosa, and by 16.2-, 7.9-, 9.5- and 7.8-fold in the intestinal mucosa. Bismuth subsalicylate exerted significant protection against DNA damage and changes in membrane microviscosity induced by the four stressors. Excellent correlations existed between the production of reactive oxygen species and the tissue damaging effects in both gastric and intestinal mucosa. In summary, the results demonstrate that physical and chemical stressors can induce gastrointestinal oxidative stress and mucosal injury through enhanced production of reactive oxygen species, and that BSS can significantly attenuate gastrointestinal injury by scavenging these reactive oxygen species.

Toxicity studies in mice of common spices, Cinnamomum zeylanicum bark and Piper longum fruits.

Acute (24 hours) and chronic (90 days) oral toxicity studies on the ethanolic extracts of common spices Cinnamomum zeylanicum Nees bark and Piper longum L. fruits were carried out in mice. Acute dosages were 0.5, 1.0 and 3 g/kg while the chronic dosage was 100 mg/kg/day. All external morphological, hematological and spermatogenic changes, in addition to body weight and vital organ weights, were recorded. The extracts of both the plants caused no significant acute or chronic mortality compared to the control during this study. During chronic treatment there was no significant change in the pre- and post treatment body weight of the test animals while the weight gain in the control group was significant. C. zeylanicum treatment caused reduction in liver weight while P. longum caused a significant increase in the weight of the lungs and spleen of the treated animals compared to the control. Hematological studies revealed a significant fall in hemoglobin level of C. zeylanicum treated animals. Both of the extracts induced a significant increase in reproductive organ weights, sperm motility, sperm count and failed to illicit any spermatotoxic effect.

Sister chromatid exchanges induced in vitro and in vivo by an extract of black pepper.

Black pepper is a spice widely used in human food. The aim of this investigation was to determine whether an alcoholic extract of the mature berries of black pepper induced genotoxic damage in vivo and in vitro. The first aspect was evaluated in mouse bone marrow cells and the second one in human lymphocytes. In both cases the rate of sister chromatid exchange (SCE) and the replicative index were determined. For the in vivo assay, ip doses of 7.0, 14.0, 28.0 and 56.0 mg/kg body weight were tested, with the following results: (1) a significant increase of SCE frequency in all doses tested compared with the control level (the highest dose produced almost a duplication of the basal rate of SCEs); (2) a similar pattern with regard to cell proliferation kinetics at all doses tested, without significant differences between them. For the in vitro assay, doses of 25.0, 50.0, 75.0 and 100.0 micrograms/ml were tested, with the following results: (1) a significant increase in the frequency of SCEs at all doses tested; a linear regression analysis of the data produced a correlation coefficient of 0.98; (2) a significant reduction in the replicative index, at the two high doses. These results demonstrated that the extract of black pepper was genotoxic in both systems.

Genotoxic effects of crude juices from Brassica vegetables and juices and extracts from phytopharmaceutical preparations and spices of cruciferous plants origin in bacterial and mammalian cells.

Crude juices of eight Brassica vegetables as well as juices and extracts of spices and phytopharmaceutical preparations from cruciferous vegetables were tested for induction of point mutations in Salmonella TA98 and TA100, repairable DNA damage in E.coli K-12 cells and clastogenic effects in mammalian cells. In bacterial assays, all juices caused genotoxic effects in the absence of metabolic activation, the ranking order being: Brussels sprouts > white cabbage > cauliflower > green cabbage > kohlrabi > broccoli > turnip > black radish. In experiments with mammalian cells, six juices induced structural chromosome aberrations. Brussels sprouts, white and green cabbage caused the strongest effects (800 microliters of juice induced a 5-fold increase over the background). In sister chromatid exchange assays, positive results were measured as well, but the effects were less pronounced. With all juices the genotoxic effects seen in mammalian cells were paralleled by a pronounced decrease in cell viability. Column fractionation experiments showed that 70-80% of the total genotoxic activity of the juices is found in the fraction which contains isothiocyanates and other breakdown products of glucosinolates, whereas phenolics and flavonoids contributed to a lesser extent to the overall effects. On the basis of these findings, and considering the negative results obtained with non-cruciferous vegetables (tomato, carrot and green pepper), it seems likely that the genotoxic effects of the juices are due to specific constituents of cruciferous plants such as glucosinolates and/or their breakdown products, in particular, isothiocyanates, which we found previously to be potent genotoxins in bacterial and mammalian cells. Finally, spices (mustards and horse radish paste) and phytopharmaceutical preparations were tested in bacterial assays. Mustards and horse radish caused very weak effects while most of the pharmaceutical preparations gave negative results, except cabbage tablets, which caused a strong and dose dependent induction of his revertants in Salmonella TA100. The present findings clearly indicate that cruciferous vegetables contain DNA damaging constituents. These observations are in contrast to earlier findings, which emphasized the antimutagenic effects of vegetable juices and also raise the question whether greatly increased consumption of Brassica vegetables or their concentrated constituents as a means for cancer prevention is indeed recommendable.