Selective inhibition of cell proliferation by lycopene in MCF-7 breast cancer cells in vitro: a proteomic analysis.

Lycopene, a red pigmented carotenoid present in many fruits and vegetables such as tomatoes, has been associated with the reduced risk of breast cancer. This study sought to identify proteins modulated by lycopene during cell proliferation of the breast cancer cell line MCF-7 to gain an understanding into its mechanism of action. MCF-7 breast cancer cells and MCF-10 normal breast cells were treated with 0, 2, 4, 6, 8, and 10 µM of lycopene for 72 h. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) tetrazolium reduction assay was used to measure cell proliferation and two-dimensional fluorescence difference gel electrophoresis to assess the changes in protein expression, which were identified using MALDI-ToF/ToF (matrix-assisted laser desorption ionization tandem time-of-flight) and Mascot database search. MTT and cell proliferation assays showed that lycopene selectively inhibited the growth of MCF-7 but not MCF-10 cells. Difference gel electrophoresis analysis revealed that proteins in the MCF-7 cells respond differently to lycopene compared with the MCF-10 cells. Lycopene altered the expression levels of proteins such as Cytokeratin 8/18 (CK8/18), CK19 and their post translational status. We have shown that lycopene inhibits cell proliferation in MCF-7 human breast cancer cells but not in the MCF-10 mammary epithelial cells. Lycopene was shown to modulate cell cycle proteins such as beta tubulin, CK8/18, CK19 and heat shock proteins.

Kyolic and Pycnogenol increase human growth hormone secretion in genetically-engineered keratinocytes.

The amount of human growth hormone (HGH) decreases significantly after the age of 30. This decrease has been implicated as one of the major causes in the signs of aging, such as thinning of the skin and bones, a decrease in lean muscle mass and an increase in adipose tissue. Supplementing the body's dwindling supply with recombinant human growth hormone (rHGH) has been shown to reverse the signs and symptoms of aging. However, drawbacks in rHGH replacement therapy include prohibitively high cost, the need for repeated injection and side effects such as carpel tunnel syndrome, gynecomastia and insulin resistance. The purpose of this study was to establish an in vitro model using genetically-engineered keratinocytes to screen natural compounds for the ability to stimulate HGH secretion. We now report that a combination of equal amounts of L-arginine and L-lysine, aged garlic extract (Kyolic), S-allyl cysteine and Pycnogenol significantly increased secretion of HGH in this in vitro model. The data indicate that this in vitro model may be used to screen for other secretagogues.

Pycnogenol reduces talc-induced neoplastic transformation in human ovarian cell cultures.

Talc and poor diet have been suggested to increase the risk of developing ovarian cancer; which can be reduced by a diet rich in fruit and vegetables. Talc is ubiquitous despite concern about its safety, role as a possible carcinogen and known ability to cause irritation and inflammation. It was recently shown that Pycnogenol (Pyc; a proprietary mixture of water-soluble bioflavonoids extracted from French maritime pine bark) was selectively toxic to established malignant ovarian germ cells. This study investigated talc-induced carcinogenesis and Pyc-induced chemoprevention. Normal human epithelial and granulosa ovarian cell lines and polymorphonuclear neutrophils (PMN) were treated with talc, or pretreated with Pyc then talc. Cell viability, reactive oxygen species (ROS) generation and neoplastic transformation by soft agar assay were measured. Talc increased proliferation, induced neoplastic transformation and increased ROS generation time-dependently in the ovarian cells and dose-dependently in the PMN. Pretreatment with Pyc inhibited the talc-induced increase in proliferation, decreased the number of transformed colonies and decreased the ROS generation in the ovarian cells. The data suggest that talc may contribute to ovarian neoplastic transformation and Pyc reduced the talc-induced transformation. Taken together, Pyc may prove to be a potent chemopreventative agent against ovarian carcinogenesis.

Suppression of LDL oxidation by garlic compounds is a possible mechanism of cardiovascular health benefit.

Hypercholesterolemia is a major risk factor for atherosclerosis, and lowering cholesterol can significantly reduce the risk for cardiovascular diseases. Oxidation of LDL has recently been recognized as playing an important role in the initiation and progression of atherosclerosis. Oxidized LDL, but not native LDL, promotes vascular dysfunction by exerting direct cytotoxicity to endothelial cells, by increasing chemotactic properties of monocytes, by transforming macrophages to foam cells, and by enhancing the proliferation of endothelial cells, monocytes, and muscle cells. All these events are recognized as contributors to cardiovascular diseases. This paper presents experimental evidence showing that several garlic compounds can suppress LDL oxidation in vitro. Short-term supplementation of garlic in human subjects has demonstrated an increased resistance of LDL to oxidation. These data suggest that suppressed LDL oxidation may be one of the mechanisms that accounts for the beneficial effects of garlic in cardiovascular health.

Kyo-Green improves sexual dysfunction in men and women.

BACKGROUND: Serendipitously, we discovered the effects of Kyo-Green, a green powdered nutritional supplement, on improving sexual dysfunctions in both men and women. In this paper, we presented two case reports and an open-labeled pilot study involving 40 subjects for a period of three months to determine the effects of Kyo-Green on sexual dysfunctions. MATERIAL/METHODS: Twenty-five men and 15 women were enrolled into this study. All the male subjects had suffered erectile dysfunction (ED). All the female subjects reported lack of libido. Subjects took one teaspoonful of Kyo-Green two times a day. Subjects completed a questionnaire at the beginning, and at the end of one, two, and three months while taking Kyo-Green. The analysis of data was based on the four questionnaires completed by the subjects. RESULTS: Increase of energy was reported by all the subjects. Sixteen males and 12 females reported satisfaction with their sex life after taking Kyo-Green for three months. Twenty of the 25 male subjects reported significant improvement in erectile dysfunction, and ability to initiate and maintain sexual activity with satisfaction. CONCLUSIONS: All the subjects experienced an increase of energy levels within a few weeks after taking this supplement. Eighty percent of the male subjects with ED regained erections while taking this supplement. The study suggests that Kyo-Green, particularly when used in conjunction with a lifestyle modification approach, may be useful in the management of sexual dysfunction in men and women who prefer a non-drug approach.

Neuroprotective effect of garlic compounds in amyloid-beta peptide-induced apoptosis in vitro.

BACKGROUND: Neuronal apoptosis is one of the pathological features of Alzheimer's disease (AD) and is associated with senile plaques containing amyloid-b peptide (Abeta). Reactive oxygen species (ROS) are proposed to be involved in the apoptotic mechanism of Abeta-mediated neurotoxicity. The purpose of this study was to determine the effects of aged garlic extract (AGE) and S-allyl cysteine (SAC) on Abeta(25-35)-induced apoptosis and ROS generation in a rat pheochromocytoma (PC12) cell line. MATERIAL/METHODS: PC12 cells were grown in RPMI 1640 medium containing 5% fetal calf serum and 10% horse serum. Cells were incubated with AGE or SAC for 24 h prior to exposure to Abeta(25-35) for various times. Cell viability, DNA fragmentation, number of apoptotic cells, caspase activity and generation of ROS were determined. RESULTS: Abeta(25-35)-induced apoptosis in PC12 cells was demonstrated by: 1) A dose-dependent loss of cell viability; 2) A time- and dose-dependent increase in apoptotic cells; 3) An induction of DNA fragmentation; and 4) An increase in caspase-3 activity and cleavage of poly (ADP-ribose) polymerase (PARP). After exposing PC12 cells to Abbeta(25-35), a significant increase in ROS preceded apoptotic events. AGE and SAC not only suppressed the generation of ROS but also attenuated caspase-3 activation, DNA fragmentation, PARP cleavage and eventually protected against Ab-induced apoptosis. CONCLUSIONS: Our data suggest that ROS may be involved in Ab-induced apoptosis in PC12 cells. They further suggest that garlic compounds can reduce apoptosis, possibly by enhancing the endogenous antioxidant defenses.

Pycnogenol as an adjunct in the management of childhood asthma.

A randomized, placebo-controlled, double-blind study involving 60 subjects, aged 6-18 years old, was conducted over a period of 3 months to determine the effect of Pycnogenol (a proprietary mixture of water-soluble bioflavonoids extracted from French maritime pine) on mild-to-moderate asthma. After baseline evaluation, subjects were randomized into two groups to receive either Pycnogenol or placebo. Subjects were instructed to record their peak expiratory flow with an Assess Peak Flow Meter each evening. At the same time, symptoms, daily use of rescue inhalers (albuterol), and any changes in oral medications were also recorded. Urine samples were obtained from the subjects at the end of the run-in period, and at 1-, 2-, and 3-month visits. Urinary leukotriene C4/D4/E4 was measured by an enzyme immunoassay. Compared with subjects taking placebo, the group who took Pycnogenol had significantly more improvement in pulmonary functions and asthma symptoms. The Pycnogenol group was able to reduce or discontinue their use of rescue inhalers more often than the placebo group. There was also a significant reduction of urinary leukotrienes in the Pycnogenol group. The results of this study demonstrate the efficacy of Pycnogenol as an adjunct in the management of mild-to-moderate childhood asthma.