The GABAA-Benzodiazepine Receptor Antagonist Flumazenil Abolishes the Anxiolytic Effects of the Active Constituents of Crocus sativus L. Crocins in Rats.

Anxiety is a chronic severe psychiatric disorder. Crocins are among the various bioactive components of the plant Crocus sativus L. (Iridaceae) and their implication in anxiety is well-documented. However, which is the mechanism of action underlying the anti-anxiety effects of crocins remains unknown. In this context, it has been suggested that these beneficial effects might be ascribed to the agonistic properties of these bioactive ingredients of saffron on the GABA type A receptor. The current experimentation was undertaken to clarify this issue in the rat. For this research project, the light/dark and the open field tests were used. A single injection of crocins (50 mg/kg, i.p., 60 min before testing) induces an anti-anxiety-like effect revealed either in the light-dark or open field tests. Acute administration of the GABAA-benzodiazepine receptor antagonist flumazenil (10 mg/kg, i.p., 30 min before testing) abolished the above mentioned anxiolytic effects of crocins. The current findings suggest a functional interaction between crocins and the GABAA receptor allosteric modulator flumazenil on anxiety.

Exploring the Valuable Carotenoids for the Large-Scale Production by Marine Microorganisms.

Carotenoids are among the most abundant natural pigments available in nature. These pigments have received considerable attention because of their biotechnological applications and, more importantly, due to their potential beneficial uses in human healthcare, food processing, pharmaceuticals and cosmetics. These bioactive compounds are in high demand throughout the world; Europe and the USA are the markets where the demand for carotenoids is the highest. The in vitro synthesis of carotenoids has sustained their large-scale production so far. However, the emerging modern standards for a healthy lifestyle and environment-friendly practices have given rise to a search for natural biocompounds as alternatives to synthetic ones. Therefore, nowadays, biomass (vegetables, fruits, yeast and microorganisms) is being used to obtain naturally-available carotenoids with high antioxidant capacity and strong color, on a large scale. This is an alternative to the in vitro synthesis of carotenoids, which is expensive and generates a large number of residues, and the compounds synthesized are sometimes not active biologically. In this context, marine biomass has recently emerged as a natural source for both common and uncommon valuable carotenoids. Besides, the cultivation of marine microorganisms, as well as the downstream processes, which are used to isolate the carotenoids from these microorganisms, offer several advantages over the other approaches that have been explored previously. This review summarizes the general properties of the most-abundant carotenoids produced by marine microorganisms, focusing on the genuine/rare carotenoids that exhibit interesting features useful for potential applications in biotechnology, pharmaceuticals, cosmetics and medicine.

Safety Assessment of Oil from Pequi (Caryocar brasiliense Camb.): Evaluation of the Potential Genotoxic and Clastogenic Effects.

Genotoxic data of medicinal plants and functional foods are required as part of the risk assessment by international regulatory agencies. Due to its food consumption and ethnopharmacological relevance, pequi oil (Caryocar brasiliense Camb.) is one of these compounds to be studied. The aim of this study was to evaluate the cytotoxic, genotoxic, and clastogenic effects of the oil from the pulp of C. brasiliense (OPCB) in vivo and in vitro. Initially, the Artemia salina in vitro assay was conducted to determine the cells viability rate of different doses of the OPCB. Subsequently, comet assay (Organization for Economic Cooperation and Development, OECD 489) and micronucleus test (OECD 474) were performed in blood and bone marrow of Wistar rats treated orally with a 125, 250, 500, or 1000 mg/kg/bw of the OPCB for 4 weeks. The chemical analysis indicated the presence of ß-carotene and lycopene in the oil. In the A. salina test, all OPCB doses maintained cell viability rates statistically similar to the negative control. The in vivo tests performed showed that OPCB did not show significant genotoxic or clastogenic effects in cells analyzed with the four doses tested. Altogether, these results indicate that, under our experimental conditions, C. brasiliense fruit oil did not reveal genetic toxicity in rat cells.

Evaluation of Cytotoxic Effect and Antioxidant Activity of Grape Seed Extract, Crocin, and Phenytoin.

Antioxidant compounds inhibit formation of free radicals, chelate catalytic metals, and scavenge free radicals in biological systems. In addition, antioxidants play a decisive role in prevention of numerous physiological dysfunctions, cancers, and metabolic disorders. This study sought to evaluate the antioxidant capacity and cytotoxic effect of grape seed extract (GSE), crocin (CRO), and phenytoin (PHEN) on a human breast cancer cell line (MCF-7). Methanol extracts of the three mentioned agents were prepared and their antioxidant activity was evaluated by diphenyl-1-picrylhydrazyl method, using Quercetine (QUER) as positive control. The 3-(4, 5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay was used to evaluate the cytotoxic effect of the extracts on Michigan Cancer Foundation-7MCF-7 cell line, using doxorubicin hydrochloride (DOX) as the positive control. Given the results, greater scavenging activity was achieved by using GSE in comparison to CRO and PHEN. Further, a significant correlation was found between the antioxidant activity and cytotoxic effects of these agents, and GSE had the highest antioxidant capacity and cytotoxic effect in comparison to CRO and PHEN.

The hazardous effects of three natural food dyes on developmental stages and longevity of Drosophila melanogaster.

Nowadays, food dyes obtained from herbal, animal, microbial and mineral sources are widely used as food additives. In this study, the toxic effects of three different natural food dyes (carmine, turmeric and annatto) on 72 ± 4 h larvae of Oregon-R wild type of Drosophila melanogaster were investigated. For this purpose, four different application doses (50, 75, 100, 125 mg mL(-1)) were chosen by means of preliminary studies. It was determined that larval mortality increased with increasing concentration in the application groups and the toxicity order was carmine > turmeric > annatto. It was observed that the survival rate was highest in the control with 98% and lowest in 125 mg mL(-1) carmine with 16%. In addition, the average lifespan of the adult individuals obtained from third instar larvae was also studied. While the average lifespan was 40.88 ± 1.44 days in the control group, these values were 10.81 ± 0.55-23.90 ± 1.27 days in the carmine group, 15.00 ± 0.80-22.42 ± 1.43 days in the turmeric group and 10.33 ± 1.03-35.68 ± 1.54 days in the annatto group, respectively. According to the obtained results, when both the developmental period from larvae into adults and the lifespan of the developing adults were compared with the control group, the food dyes were found to be toxic and the toxicity order of carmine > turmeric > annatto was identified.

Bioactivity assessment and toxicity of crocin: a comprehensive review.

Since ancient times, saffron, the dried stigma of the plant Crocus sativus L. has been extensively used as a spice and food colorant; in folk medicine it has been reputed to be efficacious for the alleviation and treatment of ailments. In addition to the three founded major constituents including crocin, picrocrocin and safranal, presence of carotenoids, carbohydrates, proteins, anthocyanins, vitamins and minerals provide valuable insights into the health benefits and nutritional value of saffron. Of the carotenoids present in saffron, highly water-soluble crocin (mono and diglycosyl esters of a polyene dicarboxylic acid, named crocetin) is responsible for the majority of its color, and appears to possess various health-promoting properties, as an antioxidant, antitumor, memory enhancer, antidepressant, anxiolytic and aphrodisiac. It is also worth noting that the crocin principle of saffron exhibited high efficacy along with no major toxicity in experimental models. We would be remiss to not consider the great potential of saffron and crocin, which benefits the cuisine and health of human life throughout the world. The present study provides a comprehensive and updated report of empirical investigations on bioactivities and biological characteristics of crocin.

Subacute toxicity assessment of carotenoids extracted from citrus peel (Nanfengmiju, Citrus reticulata Blanco) in rats.

The mixture of carotenoids extracted from citrus peel (Nanfengmiju, Citrus reticulata Blanco) was tested for subacute oral toxicity. In this study, dose levels of 0, 200, 500 and 2000 mg/kg body weight/day were administered by gavage to 10 Wistar rats/sex/group for 28 days. No statistically significant, dose-related effect on food consumption, food efficiency, body weight gain, clinical signs or ophthalmoscopic parameters was observed in any treatment group. Urinalysis, hematological, blood coagulation and serum biochemical examination as well as necropsy or histopathology showed that no observed adverse effect was found. These findings suggested that the No-Observed-Adverse-Effect Level for the mixture of carotenoids extracted from citrus peel was at least 2000 mg/kg body weight/day.

Contact sensitizing potential of annatto extract and its two primary color components, cis-bixin and norbixin, in female BALB/c mice.

The present studies were performed to examine the contact allergenic effects of an annatto extract (ANT) in female BALB/c mice. ANT at 5-10% induced a greater than threefold increase in lymph node cell proliferation when compared to the control in the LLNA. Moreover, a significant increase in the percent ear swelling at 24h after ANT challenge was observed in the MEST. A significant increase in the percentage of B cells was also observed. To determine which of the two predominant coloring components (norbixin and bixin) in ANT was responsible for the sensitizing effects of ANT, norbixin was subsequently examined, with negative results being observed in both the LLNA and MEST following treatment with norbixin (1-20%). These findings suggested that perhaps bixin was responsible for the positive responses in both the LLNA and MEST following exposure to ANT. Therefore, further studies using a partially purified cis-bixin extract were conducted. Positive responses in both the LLNA and MEST were observed in mice treated with cis-bixin at the concentrations as low as 0.1-0.5%. These results have demonstrated that cis-bixin, but not norbixin, is likely a contact sensitizer and contributes to the contact hypersensitivity effects observed following dermal exposure to ANT in mice.

Metabolites from Withania aristata with potential phytotoxic activity.

A series of apocarotenoids (1-8) and one carotenoid (9) were isolated from the leaves of Withania aristata. In addition, the tetraacetylated apocarotenoid glucosides 10-12 were obtained by acetylation, with derivative 9-hydroxymegastigma-4,6E-dien-3-one 9-O-beta-D-glucopyranoside tetraacetate (10) being described for the first time. The structures have been determined by spectroscopic and spectrometric means, mainly NMR and ESI-MS, and comparison with data reported in the literature. These metabolites were evaluated on a systematic phytotoxicity assay using the etiolated wheat coleoptile bioassay. Compounds 1-3, 9 and 12 were further assayed for their phytotoxicity on the target species Lepidium sativum, Lactuca sativa, Lycopersicum esculentum and Allium cepa. Among the assayed compounds, lutein (9) showed the most significant values for phytotoxicity, followed by the non-glycosylated apocarotenoids (6S, 9R)-vomifoliol (1) and 9-hydroxymegastigma-4,6E-dien-3-one (2).

Effect of annatto on micronuclei induction by direct and indirect mutagens in HepG2 cells.

Annatto (AN), a natural food colorant rich in carotenoids, has been reported as being an effective antioxidant, but little is known about its potential chemopreventive properties. In this study, we evaluated the ability of AN to protect human hepatoma cells (HepG2) from micronucleus (MN) induction against three different mutagens: benzo(a)pyrene (B(a)P), doxorubicin (DXR), and methyl methanesulfonate (MMS). In an attempt to clarify the possible mechanism of antimutagenicity of AN, three protocols of treatment were applied (pretreatment; simultaneous treatment, and post-treatment with AN following treatment with the mutagens). Also, cells exposed only to AN were assayed for cytotoxicity and mutagenicity. A dosage up to 10 microg/ml of AN was devoid of mutagenic activity. Protective effects were seen on micronuclei induced by B(a)P and DXR using pre and simultaneous treatment, but AN had no significant effect on MN induction by MMS in any of the protocols. Our results also show that exposure of cells to concentrations of AN higher than 10 microg/ml decreased cell viability. Taken together, our findings indicate that AN presents antimutagenic activity in vitro, but its protective effect is dependent on the mutagen and on type of treatment suggesting its potential use as a chemopreventive agent.

High dose lycopene supplementation increases hepatic cytochrome P4502E1 protein and inflammation in alcohol-fed rats.

Recent in vitro evidence suggests that the antioxidant lycopene can prevent alcohol-induced oxidative stress and inflammation. However, knowledge of possible interactions in vivo between escalating doses of lycopene and chronic alcohol ingestion are lacking. In this study, we investigated potential interactions between alcohol ingestion and lycopene supplementation and their effect on hepatic lycopene concentration, cytochrome P4502E1 (CYP2E1) induction, and inflammation. Fischer 344 rats (6 groups, n = 10 per group) were fed either a liquid ethanol Lieber-DeCarli diet or a control diet (isocaloric maltodextrin substituted for ethanol) with or without lycopene supplementation at 2 doses (1.1 or 3.3 mg x kg body weight(-1) x d(-1)) for 11 wk. Plasma and hepatic concentrations of lycopene isomers were assessed by HPLC analysis. We examined expressions of hepatic CYP2E1 and tumor necrosis factor-alpha (TNFalpha) and the incidence of hepatic inflammatory foci. Both plasma and hepatic lycopene concentrations were greater in alcohol-fed rats than in control rats supplemented with identical doses of lycopene. In contrast, alcohol-fed rats had a lower percentage of lycopene cis isomers in the plasma and the liver compared with control rats fed the same dose of lycopene. Notably, lycopene supplementation at the higher dose significantly induced hepatic CYP2E1 protein, TNFalpha mRNA, and the incidence of inflammatory foci in the alcohol-fed rats but not in the control rats. These data indicate an interaction between chronic alcohol ingestion and lycopene supplementation and suggest a need for caution among individuals consuming high amounts of both alcohol and lycopene.

Safety evaluation of a natural tomato oleoresin extract derived from food-processing tomatoes.

Experimental and epidemiological studies indicate that consumption of tomato products containing high amounts of lycopene is associated with lowered cancer risk. The protective effects of lycopene may be related to its antioxidant potential. Lycopene has been demonstrated to inhibit oxidation. A proprietary, natural tomato oleoresin extract (NTOE), is a purified tomato oleoresin containing 6% lycopene produced from tomatoes. NTOE was evaluated for toxicological effects, and found the 50% lethal dose (LD(50)), derived from the acute oral toxicity study, was greater than 5000 mg/kg body weight. The no-observed-adverse-effect level (NOAEL) derived from the 13-week study was 4500 mg/kg/day. Acute dermal toxicity study of NTOE found no toxicity at 2000 mg/kg body weight. NTOE lacked dermal irritation in the rabbit model, but was found to have moderate eye-irritant capabilities. NTOE tested at 5% (w/w) in petroleum jelly was a moderate sensitizer in the guinea pig model. There was no evidence of mutagenic potential up to 5000 microg/plate, as determined by the Ames assay. These results demonstrate the inability of NTOE to produce oral, dermal or mutagenic toxicity in animal models at doses greater than 300 times the normal human consumption of lycopene. Consumption analysis of lycopene-containing foods estimated mean daily intake of lycopene at 8.2mg/day.

A thirteen-week oral toxicity study of annatto extract (norbixin), a natural food color extracted from the seed coat of annatto (Bixa orellana L.), in Sprague-Dawley rats.

A subchronic oral toxicity study of annatto extract (norbixin), a natural food color, was conducted. Groups of 10 male and 10 female Sprague-Dawley rats were fed annatto extract at dietary levels of 0, 0.1, 0.3 and 0.9% for 13 weeks. There were no treatment-related adverse effects on body weight, food and water consumption, ophthalmology and hematology data. Blood biochemical analysis revealed changes in rats of both sexes confined to the 0.9% and 0.3% groups, including increased alkaline phosphatase, phospholipid, total protein, albumin and albumin/globulin ratio. Marked elevation in absolute and relative liver weights was also found in both sexes of the 0.9% and 0.3% groups, but not the 0.1% group. Hepatocyte hypertrophy was evident and an additional electron microscopic examination demonstrated this to be linked to abundant mitochondria after exposure to a dietary level of 0.9% annatto extract for 2 weeks. Thus, the No-Observed-Adverse-Effect-Level (NOAEL) was judged to be a dietary level of 0.1% (69 mg/kg body weight/day for males, 76 mg/kg body weight/day for females) of annatto extract (norbixin) under the present experimental conditions.

Developmental (embryo-fetal toxicity/teratogenicity) toxicity studies of synthetic crystalline lycopene in rats and rabbits.

Synthetic crystalline lycopene is a nutritional supplement to increase dietary intake of lycopene, an antioxidant carotenoid. Its potential oral developmental toxicity was studied in rats and rabbits. Each study included 3 control groups (water and matrix for Lycopene 10 CWD or LycoVit 10%), 3 Lycopene 10 CWD groups [500, 1500 and 3000 (rats)/2000 (rabbits) mg/kg/day] and 1 LycoVit 10% group [3000 mg/kg/day (rats)/2000 (rabbits)]. The high dosages were at maximum achievable concentrations and dosage volumes (15 and 10 ml/kg for rats and rabbits, respectively) of the highly viscous test material suspensions. Dosages were administered on gestation days (GDs) 6 through 19 (rats) or GDs 6 through 28 (rabbits). Endpoints evaluated included viability, body weight, feed consumption, necropsy observations [GD 20 (rats)/GD 29 (rabbits)], uterine contents and fetal viability, gender, body weight and morphology (skeletons double-stained). Feed consumption and weight gain were essentially unaffected in rats and rabbits, despite intubation problems in both species and reduced gastrointestinal motility and mortality in rabbits attributable to the physical properties of the gels. Neither Lycopene 10 CWD nor LycoVit 10% caused direct maternal or developmental toxicity in rats or rabbits at dosages as high as 3000 or 2000 mg/kg/day, respectively.

Summary of safety studies conducted with synthetic lycopene.

Lycopene belongs to the group of natural carotenoids, which are found in many fruits and vegetables, but predominantly in tomatoes and tomato-based products. This manuscript summarizes the safety of synthetic lycopene as a water-dispersible beadlet formulation containing antioxidants and includes acute and subchronic safety studies, reproductive studies, genotoxicity studies, metabolic studies, and exploratory studies on the hepatic uptake of lycopene. Lycopene has a low order of acute toxicity and no significant toxicity has been observed in rats treated with lycopene beadlet formulations in the diet at doses of up to 500 mg/kg bw/day for 14 weeks or 1000 mg/kg bw/day for 4 weeks. No teratogenic effects were noted in a rat two-generation study (1000 ppm in the diet) or in a teratology study in rats with 1000 mg/kg bw/day lycopene as beadlet formulations. Lycopene accumulates in hepatocytes and to a lesser extent in spleen. In short-term studies with synthetic lycopene, as a beadlet formulation, and natural source lycopene, as tomato concentrate, the accumulation of lycopene in the liver and the presence of pigment deposits in the hepatocytes were similar and neither was associated with any histopathological changes. The pigment deposits in hepatocytes are no longer present after approximately 13 weeks of depletion, demonstrating reversibility for this effect. Unformulated pure crystalline lycopene and lycopene as a 10% beadlet formulation are not genotoxic as determined in a comprehensive battery of tests, however, improperly stored, unformulated crystalline lycopene can degrade to mutagenic products if exposed to light and air. Lycopene is commercially available only in formulated forms, containing antioxidants, which prevent the degradation of lycopene and other excipients that provide for water dispersibility. In the animal studies, there is a large margin of safety based on the repeated dose safety and reproductive/teratology studies in rodents. In humans, there is a very long history of use with respect to dietary exposure, and even in the case of very high exposures from dietary sources, there is no indication of any significant adverse effects.

Ninety-day oral toxicity study of lycopene from Blakeslea trispora in rats.

Lycopene, as a suspension in sunflower oil (20% w/w), was tested for subchronic toxicity by administration at dietary concentrations of 0, 0.25, 0.50, and 1.0% to groups of 20 male and 20 female Wistar rats for a period of 90 days. The lycopene examined in this study was derived from a fungal biomass (Blakeslea trispora). Lycopene intake was calculated to be 0, 145, 291, and 586mg/kg body weight/day in control through high-dose males and 0, 156, 312, and 616mg/kg body weight/day in control through high-dose females. The results from this study do not provide any evidence of toxicity of lycopene at dietary levels up to 1.0% as demonstrated by the findings of clinical observations, neurobehavioral observations, motor activity assessment, body weight and food consumption measurements, ophthalmoscopic examinations, hematology, clinical chemistry, urinalysis, organ weights, gross pathology, or histopathology. The No-Observed-Effect Level (NOEL) was 1.0% in the diet, the highest dietary concentration tested.

Thirteen-week oral toxicity study of synthetic lycopene products in rats.

Synthetic crystalline lycopene provides an alternative to extracts of naturally occurring lycopene for use in dietary supplements and functional foods. BASF Lycopene 10 CWD and Lyco Vit 10% formulated products each contain approximately 10% synthetic lycopene. These products were evaluated for toxicological and behavioral effects during a 13-week oral dosing study with male and female Wistar rats. Doses of 0, 500, 1500 and 3000 mg/kg body weight/day Lycopene CWD and 3000 mg/kg body weight /day Lyco Vit, as well as 3000 mg/kg body weight /day of the matrices used to formulate and stabilize each product, were administered by gavage to 10 rats/sex/day. A satellite group of five rats/sex received 0 or 3000 mg/kg body weight /day of each formulated product for an interim evaluation at 4 weeks of feeding. No statistically significant, dose-related effects on body weight, body weight gain, food consumption, hematology, urinalysis, clinical chemistry or ophthalmoscopic parameters were seen in any of the lycopene product or lycopene formulation matrix groups in comparison to the vehicle control group after 4 or 13 weeks of dosing. No deaths attributed to the test articles occurred during the study and the only clinical finding and at necropsy was the presence of red pigment in the feces and gastrointestinal tract that was associated with the red-pigmented test materials. No significant or dose-related abnormalities were found at necropsy or in microscopic evaluations of tissues collected at termination. Rats evaluated in home cages or in open field tests for behavioral and sensorimotor effects during the final week of the study showed no signs of treatment-related effects. The no-observed-adverse-effect level (NOAEL) for this study was concluded to be 3000 mg/kg body weight/day for both Lycopene CWD and Lyco Vit. The results of this study thus demonstrate the absence of any significant toxicological findings with Lycopene CWD and Lyco Vit products even at very high dose levels.

Genotoxicity of gardenia yellow and its components.

Gardenia fruit (Gardenia jasminoides ELLIS) is widely used as a natural food colorant and as a traditional Chinese medicine for treatment of hepatic and inflammatory diseases. "Gardenia yellow" is a natural food colorant which is extracted by ethanol from gardenia fruit. The purpose of this study was to evaluate the genotoxicity of gardenia yellow. Genotoxicity of gardenia yellow and its components, crocetin, gentiobiose (a component of crocin), geniposide and genipin (formed by hydrolysis of geniposide), was studied by Ames test, rec-assay, and sister chromatid exchange (SCE) using V79 cells. Gardenia yellow and its components were found not to be mutagenic in the Salmonella reverse mutation assay. Gardenia yellow and genipin caused damage of DNA in rec-assay. Gardenia yellow induced a significant dose-dependent increase of SCE frequency (8.6 times at 1000 microg/ml as the value for the solvent control). Only genipin induced SCEs significantly among the components of gardenia yellow. Moreover, genipin induced a significant increase of tetraploids at all doses tested (95% at 8 microg/ml). Gardenia yellow preparation was analyzed by capillary electrophoresis (CE), and geniposide was detected. However, genipin was not observed. In conclusion, we have shown that genipin possesses genotoxicity. Furthermore, there were unidentified genotoxicants in gardenia yellow.

Membrane associated antitumor effects of crocine-, ginsenoside- and cannabinoid derivates.

In the present work a systematic study was initiated with crocine, ginsenoside and cannabinoid derivatives on multidrug resistant mouse lymphoma cells, viral tumor antigen expression and some human leukocyte functions. Among saffron derivatives, crocin and picrocrocin, triglucosyl and diglucosyl crocetin were ineffective on the reversal of multidrug resistance of lymphoma cells. Ginsenoside increased drug accumulation and tumor antigen expression at 2.0-20.0 micrograms/mL. Some cannabinoid derivatives such as cannabinol, cannabispirol and cannabidiol increased drug accumulation, while cannabidiolic acid, delta-9-THC and tetrahydro-cannabidiolic acid reduced drug accumulation of the human mdr1-gene transfected mouse lymphoma cells. The reversal of multidrug resistance is the result of the inhibition of the efflux pump function in the tumor cells. Crocetin esters were less potent than crocin itself in the inhibition of EBV early antigen expression. However crocin and diglucosylcrocetin inhibited early tumor antigen expression of adenovirus infected cells, but triglucosylcrocetin was less effective at 0.01-1.0 microgram/mL. The crocin had no antiviral effect [on HSV-2 infected vero cells] up to 25 micrograms/mL concentration. Ginsenosides had a moderate inhibitory effect except ginsenoside Rb1 (was the less effective) on the drug efflux pump. Among the cannabinoid derivatives the cannabinol and cannabispirol increased drug accumulation, while cannabidiolic acid and delta-8-THC, delta-9-THC and tetrahydro-cannabinol reduced drug accumulation in multidrug resistant mouse lymphoma cells. It is interesting that ginsenosides had a chemical structure-dependent immunomodulating effect by enhancing the activity of NK-cells and ADCC activities.