Antimutagenic, antitumor and estrogen receptor binding activity of the rare plant Shortia galacifolia: An ethnobotanical and chemosystematic approach.

OBJECTIVE: Shortia and other members of the Diapensiaceae family have ethnomedicinal history in both Eastern and Western hemispheres. Based on ethnopharmacological and chemosystematic evidence, pharmacological and toxicological bioassays were conducted on the rare plant Oconee Bell, Shortia galacifolia. MATERIALS AND METHODS: Extracts were examined in assays for antimutagenicity, antitumor and estrogen receptor (ER)-binding activity. Antitumor activity was assessed by the tumor induction assay (TiA), using Agrobacterium tumefaciens based on its ability to transform plant tissue. Antimutagenicity was examined using the Ames bacterial reverse mutation test. Recombinant human ERα and ERß proteins were utilized to screen extracts for receptor selectivity. RESULTS: All concentrations of extracts inhibited A. tumefaciens-induced tumor formation on potato discs, with the mature rhizome extracts having the most marked inhibition. All three plant extracts significantly inhibited the formation of histidine-independent revertant colonies after exposure to the mutagen 2-aminoanthracene (2-AA) in the Ames Salmonella mutagenicity assay. In the ER binding assays, ERß, but not ERα, displayed affinity for Shortia extracts. CONCLUSION: Antitumor, ER binding and antimutagenic activities of S. galacifolia extracts were identified using rapid bench-top assays and warrant further investigations.

Effects of Panax ginseng, zearalenol, and estradiol on sperm function.

BACKGROUND: Estrogen signaling pathways are modulated by exogenous factors. Panax ginseng exerts multiple activities in biological systems and is classified as an adaptogen. Zearalenol is a potent mycoestrogen that may be present in herbs and crops arising from contamination or endophytic association. The goal of this study was to investigate the impact of P. ginseng, zearalenol and estradiol in tests on spermatozoal function. METHODS: The affinity of these compounds for estrogen receptor (ER)-alpha and beta (ERα and ERß)-was assessed in receptor binding assays. Functional tests on boar spermatozoa motility, movement and kinematic parameters were conducted using a computer-assisted sperm analyzer. Tests for capacitation, acrosome reaction (AR), and chromatin decondensation in spermatozoa were performed using microscopic analysis. RESULTS: Zearalenol-but not estradiol (E2)- or ginseng-treated spermatozoa-decreased the percentage of overall, progressive, and rapid motile cells. Zearalenol also decreased spontaneous AR and increased chromatin decondensation. Ginseng decreased chromatin decondensation in response to calcium ionophore and decreased AR in response to progesterone (P4) and ionophore. CONCLUSION: Zearalenol has adverse effects on sperm motility and function by targeting multiple signaling cascades, including P4, E2, and calcium pathways. Ginseng protects against chromatin damage and thus may be beneficial to reproductive fitness.

Multiple Integrated Complementary Healing Approaches: Energetics & Light for bone.

A synergistic-healing strategy that combines molecular targeting within a system-wide perspective is presented as the Multiple Integrated Complementary Healing Approaches: Energetics And Light (MICHAEL). The basis of the MICHAEL approach is the realization that environmental, nutritional and electromagnetic factors form a regulatory framework involved in bone and nerve healing. The interactions of light, energy, and nutrition with neural, hormonal and cellular pathways will be presented. Energetic therapies including electrical, low-intensity pulsed ultrasound and light based treatments affect growth, differentiation and proliferation of bone and nerve and can be utilized for their healing benefits. However, the benefits of these therapies can be impaired by the absence of nutritional, hormonal and organismal factors. For example, lack of sleep, disrupted circadian rhythms and vitamin-D deficiency can impair healing. Molecular targets, such as the Wnt pathway, protein kinase B and glucocorticoid signaling systems can be modulated by nutritional components, including quercetin, curcumin and Mg(2+) to enhance the healing process. The importance of water and water-regulation will be presented as an integral component. The effects of exercise and acupuncture on bone healing will also be discussed within the context of the MICHAEL approach.

Identification of kinases, phosphatases, and phosphorylation sites in human and porcine spermatozoa.

Multiple inter-connected signaling pathways, involving kinases and phosphatases, form a framework that controls sperm motility, function, and fertilizing ability. Methods that give a broad view of the proteomic landscape may prove valuable in uncovering new crosstalk connections, as well as in discovering new proteins within this regulatory framework. A multi-immunoblotting strategy was utilized to evaluate this concept on human and porcine spermatozoa samples. In human and porcine spermatozoa, a diversity of kinases were identified including protein kinase A (PKA), protein kinase B (PKB), isoforms of protein kinase C (PKC), calmodulin-dependent kinases (CAMK), casein kinase (CK), and isoforms of glycogen synthase kinase (GSK3). Several phosphatases, such as protein phosphatase (PP)-1, PP2A, PP2C, and mitogen activated protein kinase (MAPK) phosphatase (MKP-1), were identified in human spermatozoa. The phosphorylation epitopes recognized belonged to members of the MAPK family, in addition to α and ß isoforms of GSK3 and cAMP response element binding protein (CREB). Proteomic approaches that allow a broad view may aid in understanding the crosstalk between signaling systems in spermatozoal physiology.

Impact of kudzu and puerarin on sperm function.

The goal of this study was to investigate the impact of kudzu (Pueraria mirifica) and the isoflavone puerarin in functional toxicological tests on spermatozoa and to assess the affinity of extracts and pure isoflavones for estrogen receptor (ER)-alpha and -beta (ERα, ERß) in receptor binding assays. Capacitation, acrosome reaction and chromatin decondensation in spermatozoa were analyzed using microscopic analysis. Kudzu, but not puerarin, reduced motility of sperm. Puerarin reduced the percent spontaneous acrosome reaction in spermatozoa. The pathways used by kudzu that affect sperm function are not fully mirrored by puerarin. Puerarin, kudzu and its other phytoestrogenic components displayed preferential affinity for ERß, however the diverse effects of kudzu and puerarin on sperm function implicate the involvement of multiple signaling systems.

Composition, mosquito larvicidal, biting deterrent and antifungal activity of essential oils of different plant parts of Cupressus arizonica var. glabra ('Carolina Sapphire').

Essential oils obtained by hydrodistillation of female cones (FC), male cones (MC), needle-twigs (NT) and wood-bark (WB) of 'Carolina Sapphire' [Cupressus arizonica var glabra (Sudw.) Little] were analyzed simultaneously by gas chromatography (GC) and gas chromatography-mass spectrometry (GC/MS). The main constituents of the investigated essential oils were as follows: FC: alpha-pinene 68.5% and myrcene 11.9%; MC: alpha-pinene 22.5%, epi-zonarene 9.9%, zonarene 6.9%, limonene 6.8% and cadina-3,5-diene 6.1%; NT: alpha-pinene 20.7%, epi-zonarene 10.4%, zonarene 7.6%, limonene 5.9% umbellulone 5.8% and cadina-3,5-diene 5.8%; WB: alpha-pinene 40.7%, limonene 3.2% and umbellulone 2.9%. Essential oils were evaluated for antifungal activity against the strawberry anthracnose causing fungal plant pathogens Colletotrichum acutatum, C. fragariae and C. gloeosporioides. Only WB essential oil showed moderate activity when compared with the commercial antifungal standards. Carolina Sapphire essential oils exhibited biting deterrent activity at 10 microg/cm2 with Biting Deterrence Index (BDI) values of 0.64, 0.59, 0.65 and 0.72, for FC, MC, NT and WB, respectively, and were significantly lower (P < or = 0.05) than the synthetic insect repellent, DEET (N,N-diethyl-meta-toluamide), at 25 nmol/cm2 against Aedes aegypti. The biting deterrence of NT and WB oils at 100 microg/cm2 with BDI values of 1.04 and 1.01, respectively, were similar, whereas the activity of MC (BDI = 0.88) and FC (BDI = 0.62) essential oils was lower than DEET at 25 nmol/cm2. Essential oil from FC with LD50 of 33.7 ppm was most toxic against 1-day-old Ae. aegypti larvae at 24-h post treatment. Female cone essential oil was significantly more toxic than MC (LD50 = 53.6 ppm), NT (LD50 = 55.5 ppm) and WB (LD50 = 44.6 ppm) essential oils.

Mycotoxins in root extracts of American and Asian ginseng bind estrogen receptors alpha and beta.

The estrogenic activity of ginseng has been the subject of conflicting reports. Cell proliferation, induction of estrogen-responsive genes, and isolated cases of adverse reactions such as postmenopausal vaginal bleeding and gynecomastia have been reported after ginseng treatment. Other studies report antiproliferative effects with no induction of estrogen-responsive genes. We developed estrogen receptor (ER) alpha and ER alpha competitive binding assays using recombinant receptors and [(3)H]-17 alpha-estradiol to detect phytoestrogens in extracts of Asian ginseng root (Panax ginseng C. A. Meyer) and American ginseng root (Panax quinquefolius L.). Root extracts contained substances that bound both receptor isoforms. These substances had a two to three times greater affinity for ER alpha. Significantly higher binding was found in methanol extracts than in hot water extracts. Subsequent analysis of the extracts revealed significant ER binding attributable to zearalenone, the estrogenic mycotoxin produced by several Fusarium species. The ER showed no binding affinity for Rb1 and Rg1, the major ginsenosides found in P. quinquefolius and P. ginseng, respectively. Thus, ginseng extraction methods, plant species tested, and mycotoxin contaminants may help to explain the disparate literature reports. The prevalence and health significance of fungal contamination in herbal products used for medicinal purposes should be further investigated.