RESUMEN

In a previous in vitro study, the standardized turmeric extract, HSS-888, showed strong inhibition of Aß aggregation and secretion in vitro, indicating that HSS-888 might be therapeutically important. Therefore, in the present study, HSS-888 was evaluated in vivo using transgenic 'Alzheimer' mice (Tg2576) over-expressing Aß protein. Following a six-month prevention period where mice received extract HSS-888 (5mg/mouse/day), tetrahydrocurcumin (THC) or a control through ingestion of customized animal feed pellets (0.1% w/w treatment), HSS-888 significantly reduced brain levels of soluble (∼40%) and insoluble (∼20%) Aß as well as phosphorylated Tau protein (∼80%). In addition, primary cultures of microglia from these mice showed increased expression of the cytokines IL-4 and IL-2. In contrast, THC treatment only weakly reduced phosphorylated Tau protein and failed to significantly alter plaque burden and cytokine expression. The findings reveal that the optimized turmeric extract HSS-888 represents an important step in botanical based therapies for Alzheimer's disease by inhibiting or improving plaque burden, Tau phosphorylation, and microglial inflammation leading to neuronal toxicity.

Asunto(s)

Enfermedad de Alzheimer/tratamiento farmacológico , Enfermedad de Alzheimer/metabolismo , Péptidos beta-Amiloides/metabolismo , Antioxidantes/uso terapéutico , Extractos Vegetales/uso terapéutico , Proteínas tau/metabolismo , Enfermedad de Alzheimer/genética , Precursor de Proteína beta-Amiloide/genética , Amiloidosis/tratamiento farmacológico , Análisis de Varianza , Animales , Antioxidantes/farmacología , Curcuma , Citocinas/metabolismo , Modelos Animales de Enfermedad , Ensayo de Inmunoadsorción Enzimática , Humanos , Ratones , Ratones Transgénicos , Mutación/genética , Fragmentos de Péptidos/metabolismo , Fosforilación/efectos de los fármacos , Extractos Vegetales/farmacología

RESUMEN

The present study explored the prophylactic and restorative benefits of cacao and red sage using both in vitro and in vivo models of stroke. For the in vitro study, we initially exposed primary rat cells to the established oxygen-glucose deprivation (OGD) stroke model followed by reperfusion under normoxic conditions, then added different cacao and sage concentrations to the cell culture media. Trypan blue cell viability results revealed specific cacao and sage dosages exerted significant therapeutic effects against OGD-induced cell death compared to cultured cells treated with extract vehicle. We next embarked on testing the therapeutic effects of cacao and sage in an in vivo model of stroke when extract treatment commenced either prior to or after transient middle cerebral artery occlusion (MCAo). Significant reduction in ischemic cell death within the peri-infarct area coupled with better performance in routine motor and neurological tasks were demonstrated by stroke animals that received cacao or sage extracts prior to MCAo compared to those that received the extracts or vehicle after MCAo. In summary, the present results demonstrate that neuroprotective effects were afforded by plant extract treatment, and that the in vitro stroke paradigm approximates in vivo efficacy when considering prophylactic treatment for stroke.

RESUMEN

Inhibition of beta-amyloid (A beta) accumulation and A beta fibril (fA beta) formation from A beta are attractive therapeutic targets for the treatment of Alzheimer's disease (AD). While previous studies have shown anti-amyloidogenic effects of curcumin in vitro and in vivo, no studies have examined optimized turmeric extracts enriched in curcuminoids or turmerones. Three standardized turmeric extracts, HSS-838, HSS-848, and HSS-888, were prepared with different chemical profiles to investigate their potential therapeutic benefits for AD. These extracts were fingerprinted by DART TOF-MS to reveal the significant chemical complexity. In addition four curcuminoids (curcumin, tetrahydrocurcumin, demethoxycurcumin and bisdemethoxycurcumin) were also examined. We measured the effects of the extracts and curcuminoids, on the aggregation of A beta by using a thioflavin T cell-free assay and the secretion of A beta from human neuronal cells (SweAPP N2A cells) in vitro. All three extracts and the curcuminoids showed dose-dependent inhibition of fA beta aggregation from A beta(1-42) in the cell-free assay, with IC(50) values of

Asunto(s)

Péptidos beta-Amiloides/metabolismo , Curcumina/farmacología , Extractos Vegetales/farmacología , Antiinflamatorios/farmacología , Línea Celular , Medios de Cultivo Condicionados , Curcuma , Curcumina/análogos & derivados , Diarilheptanoides , Ensayo de Inmunoadsorción Enzimática , Humanos , Espectrometría de Masas , Fitoterapia

RESUMEN

A ionization technique in mass spectrometry called Direct Analysis in Real Time Mass Spectrometry (DART TOF-MS) coupled with a Direct Binding Assay was used to identify and characterize anti-viral components of an elderberry fruit (Sambucus nigra L.) extract without either derivatization or separation by standard chromatographic techniques. The elderberry extract inhibited Human Influenza A (H1N1) infection in vitro with an IC(50) value of 252+/-34 microg/mL. The Direct Binding Assay established that flavonoids from the elderberry extract bind to H1N1 virions and, when bound, block the ability of the viruses to infect host cells. Two compounds were identified, 5,7,3',4'-tetra-O-methylquercetin (1) and 5,7-dihydroxy-4-oxo-2-(3,4,5-trihydroxyphenyl)chroman-3-yl-3,4,5-trihydroxycyclohexanecarboxylate (2), as H1N1-bound chemical species. Compound 1 and dihydromyricetin (3), the corresponding 3-hydroxyflavonone of 2, were synthesized and shown to inhibit H1N1 infection in vitro by binding to H1N1 virions, blocking host cell entry and/or recognition. Compound 1 gave an IC(50) of 0.13 microg/mL (0.36 microM) for H1N1 infection inhibition, while dihydromyricetin (3) achieved an IC(50) of 2.8 microg/mL (8.7 microM). The H1N1 inhibition activities of the elderberry flavonoids compare favorably to the known anti-influenza activities of Oseltamivir (Tamiflu; 0.32 microM) and Amantadine (27 microM).

Asunto(s)

Antivirales/química , Antivirales/farmacología , Flavonoides/metabolismo , Flavonoides/farmacología , Subtipo H1N1 del Virus de la Influenza A/efectos de los fármacos , Infecciones por Orthomyxoviridae/prevención & control , Sambucus/química , Animales , Antivirales/uso terapéutico , Línea Celular , Perros , Flavonoides/química , Humanos , Gripe Humana/prevención & control

RESUMEN

BACKGROUND: The development of antiviral drugs has provided crucial new means to mitigate or relieve the debilitating effects of many viral pathogens. Regular use of these drugs has led to generation of resistant strains, making the control of many viral infections very difficult, particularly in HIV-seropositive and AIDS patients. A rich source for the discovery of new HIV infection inhibitors has been, and continues to be, the 'mining' of the large diversity of compounds already available in nature, and specifically those from botanical extracts. METHODS: Using a newly developed direct binding assay with mass spectrometry technology (direct analysis in real-time time-of-flight mass spectrometry), we were able to show that compounds present in extracts of elderberry, cinnamon and green tea bind to and block HIV type-1 (HIV-1) infection in target cells. RESULTS: The compounds that blocked HIV-1 infection were flavonoids and A-type proanthocyanidins. The 50% inhibitory concentration values of these extracts ranged from 0.5 to 201 microg/ml for four different HIV-1 serotypes. Interaction matrices with the elderberry extract and enfuvirtide, a peptide HIV-1 fusion inhibitor, revealed significant super additive effects. This indicates that the compounds in elderberry that prevent HIV-1 infection are likely to bind to viral glycoproteins other than gp41 (the binding site for enfuvirtide). CONCLUSIONS: Optimized elderberry, green tea and cinnamon extracts rich in certain flavonoid compounds were shown to block HIV-1 entry and infection in GHOST cells. As such, these types of botanical extracts could provide a starting point for the development of possible safe and reliable cotherapies for HIV-1-positive individuals, as well as for the identification of new small molecules as leading drug candidates for HIV-1 therapeutics and microbicides.

Asunto(s)

Fármacos Anti-VIH/farmacología , VIH-1/efectos de los fármacos , VIH-1/fisiología , Extractos Vegetales/farmacología , Internalización del Virus/efectos de los fármacos , Fármacos Anti-VIH/química , Fármacos Anti-VIH/metabolismo , Fármacos Anti-VIH/toxicidad , Sitios de Unión , Camellia sinensis/química , Catequina/análogos & derivados , Catequina/farmacología , Línea Celular Tumoral , Cinnamomum zeylanicum/química , Relación Dosis-Respuesta a Droga , Descubrimiento de Drogas , Sinergismo Farmacológico , Enfuvirtida , Proteína gp41 de Envoltorio del VIH/farmacología , Infecciones por VIH/prevención & control , VIH-1/metabolismo , Humanos , Espectrometría de Masas , Fragmentos de Péptidos/farmacología , Extractos Vegetales/química , Extractos Vegetales/metabolismo , Extractos Vegetales/toxicidad , Reproducibilidad de los Resultados , Sambucus/química , Factores de Tiempo , Virión/efectos de los fármacos , Virión/metabolismo , Virión/fisiología

RESUMEN

Rice bran, the outer bran and germ of the kernel and a by-product of rice milling, is rich in phytonutrients but has been underutilized because of lipid content instability. New methods for the processing of rice bran have yielded a stabilized form that is increasingly used in foods and dietary supplements. Recent studies have documented a role for stabilized rice bran (SRB) in treating diabetes and arthritis, although little is known of the bioactive compounds that impart these health benefits. Here we characterize the chemical composition of three extracts of SRB and identify the functional bioactives contributing to the inhibitory properties against three key pro-inflammatory enzymes (cyclooxygenase [COX] 1, COX2, and 5-lipoxygenase [5-LOX]) that control the inflammatory cascade involved in impaired joint health, pain, and arthritis. One extract (SRB-AI) demonstrated significant COX1 and COX2 inhibitory activities with 50% inhibitory concentration (IC(50)) values for COX1 and COX2 of 305 and 29 microg/mL, respectively, but no 5-LOX inhibition. The second extract (SRB-AII) inhibited COX1, COX2, and 5-LOX with IC(50) values of 310, 19, and 396 microg/mL, respectively. The third extract (SRB-AIII), a blend of SRB-AI and SRB-AIII, inhibited COX1, COX2, and 5-LOX with respective IC(50) values of 48, 11, and 197 microg/mL. Analysis of the extracts by direct analysis in real time time of flight-mass spectrometry revealed that SRB-AI, SRB-AII, and SRB-AIII contain over 620, 770, and 810 compounds, respectively. Of these, 17 were identified as key bioactives for COX and/or LOX inhibition. These SRB extracts have applications for functional foods and dietary supplements for control of inflammation and joint health.

Asunto(s)

Antiinflamatorios/farmacología , Inhibidores de la Ciclooxigenasa/farmacología , Inhibidores de la Lipooxigenasa/farmacología , Oryza/química , Extractos Vegetales/farmacología , Antiinflamatorios/química , Antiinflamatorios/aislamiento & purificación , Inhibidores de la Ciclooxigenasa 2/química , Inhibidores de la Ciclooxigenasa 2/aislamiento & purificación , Inhibidores de la Ciclooxigenasa 2/farmacología , Inhibidores de la Ciclooxigenasa/química , Inhibidores de la Ciclooxigenasa/aislamiento & purificación , Relación Dosis-Respuesta a Droga , Inhibidores de la Lipooxigenasa/química , Inhibidores de la Lipooxigenasa/aislamiento & purificación , Extractos Vegetales/química , Extractos Vegetales/aislamiento & purificación , Semillas

RESUMEN

A nettle (Urtica dioica) extract shows in vitro inhibition of several key inflammatory events that cause the symptoms of seasonal allergies. These include the antagonist and negative agonist activity against the Histamine-1 (H(1)) receptor and the inhibition of mast cell tryptase preventing degranulation and release of a host of pro-inflammatory mediators that cause the symptoms of hay fevers. The nettle extract also inhibits prostaglandin formation through inhibition of Cyclooxygenase-1 (COX-1), Cyclooxygenase-2 (COX-2), and Hematopoietic Prostaglandin D(2) synthase (HPGDS), central enzymes in pro-inflammatory pathways. The IC(50) value for histamine receptor antagonist activity was 251 (+/-13) microg mL(-1) and for the histamine receptor negative agonist activity was 193 (+/-71) microg mL(-1). The IC(50) values for inhibition of mast cell tryptase was 172 (+/-28) microg mL(-1), for COX-1 was 160 (+/-47) microg mL(-1), for COX-2 was 275 (+/-9) microg mL(-1), and for HPGDS was 295 (+/-51) microg mL(-1). Through the use of DART TOF-MS, which yields exact masses and relative abundances of compounds present in complex mixtures, bioactives have been identified in nettle that contribute to the inhibition of pro-inflammatory pathways related to allergic rhinitis. These results provide for the first time, a mechanistic understanding of the role of nettle extracts in reducing allergic and other inflammatory responses in vitro.

Asunto(s)

Antagonistas de los Receptores Histamínicos/farmacología , Mediadores de Inflamación/farmacología , Extractos Vegetales/farmacología , Rinitis Alérgica Perenne/tratamiento farmacológico , Urtica dioica/química , Línea Celular , Inhibidores de la Ciclooxigenasa/farmacología , Humanos , Oxidorreductasas Intramoleculares/antagonistas & inhibidores , Lipocalinas/antagonistas & inhibidores , Espectrometría de Masas , Triptasas/antagonistas & inhibidores

RESUMEN

ABSTRACT To explore the potential for nontoxic crop protection technologies based on the inhibition of fungal spore adhesion, we have tested the effect of synthetic zosteric acid (p-(sulfo-oxy) cinnamic acid), a naturally occurring phenolic acid in eelgrass (Zostera marina L.) plants, on spore adhesion and infection in two pathosystems: rice blast caused by Magnaporthe grisea and bean anthracnose caused by Colletotrichum lindemuthianum. We have shown that zosteric acid inhibits spore adhesion to model and host leaf surfaces and that any attached spores fail to develop appressoria, and consequently do not infect leaf cells. Low concentrations of zosteric acid that are effective in inhibiting adhesion are not toxic to either fungus or to the host. The inhibition of spore adhesion in the rice blast pathogen is fully reversible. On plants, zosteric acid reduced (rice) or delayed (bean) lesion development. These results suggest that there is potential for novel and environmentally benign crop protection technologies based on manipulating adhesion.

RESUMEN

The unusual appearance of a commensal eelgrass limpet [Tectura depicta (Berry)] from southern California at high density (up to 10 shoot-1) has coincided with the catastrophic decline of a subtidal Zostera marina L. meadow in Monterey Bay, California. Some commensal limpets graze the chloroplast-rich epidermis of eelgrass leaves, but were not known to affect seagrass growth or productivity. We evaluated the effect on eelgrass productivity of grazing by limpets maintained at natural densities (8±2 shoot-1) in a natural light mesocosm for 45 days. Growth rates, carbon reserves, root proliferation and net photosynthesis of grazed plants were 50-80% below those of ungrazed plants, but biomass-specific respiration was unaffected. The daily period of irradiance-saturated photosynthesis (H sat) needed to maintain positive carbon balance in grazed plants approached 13.5 h, compared with 5-6 h for ungrazed plants. The amount of carbon allocated to roots of ungrazed plants was 800% higher than for grazed plants. By grazing the chlorophyll-rich epidermis, T. depicta induced carbon limitation in eelgrass growing in an other-wise light-replete environment. Continued northward movement of T. depicta, may have significant impacts on eelgrass production and population dynamics in the northeast Pacific, even thought this limpet consumes very little plant biomass. This interaction is a dramatic example of top-down control (grazing/predation) of eelgrass productivity and survival operating via a bottom-up mechanism (photosynthesis limitation).

RESUMEN

Photosynthetic responses of the temperate seagrass, Zostera marina L., were examined by manipulations of photon flux density in an eelgrass bed in Great Harbor, Woods Hole, MA during August 1981. Sun reflectors and light shading screens were placed at shallow (1.3 m) and deep (5.5 m) stations in the eelgrass bed to increase (+35% to +40%) and decrease (-55%) ambient photon flux densities. The portion of the day that light intensities exceeding the light compensation point for Z. marina (H comp) and the light saturation point (H sat) were determined to assess the impact of the reflectors and shades. The H comp and H sat periods at the deep station shading screen were most strongly affected; H comp was reduced by 11% and H sat was reduced by 52%. Light-saturated photosynthetic rates, dark respiration rates, leaf chlorophyll content, chlorophyll a/b, PSUO2 size, PSU density, leaf area, specific leaf area, leaf turnover times and leaf production rates were determined at the end of three sets of 1- to 2-week experiments. None of the measured parameters were affected by the photon flux density manipulations at the shallow station; however, at the deep station leaf production rates were significantly reduced under the shading screen and chlorophyll a/b ratios were higher at the reflector. These results indicate that adjustment to short-term changes in light regime in Z. marina is largely by leaf production rates. Further, the most dramatic changes in the periods of compensating or saturating photon flux densities had the greatest impact on the measured photosynthetic responses.