Biomedical subjects

Mary Ann Lila

Publications and source records attributed to Mary Ann Lila.

16 recordsLinked to original sources

PubMed2006

ent-Beyerane diterpenoids from the heartwood of Excoecaria parvifolia.

Chromatographic fractionations of the toluene extract of the heartwood of Excoecaria parvifolia collected in Australia resulted in the isolation of 12 beyerane diterpenes (1-12), and the triterpene, lupeol. Four of the isolated diterpenoids (5-7 and 12) have unusual structures: ent-3-oxa-beyer-15-en-2-one, (5); ent-15,16-epoxy-2-hydroxy-19-norbeyer-1,4-dien-3-one (6); methyl ent-2,4-seco-15,16-epoxy-4-oxo-3,19-dinorbeyer-15-en-2-oate (7); and ent-2,17-dihydroxy-19-norbeyer-1,4,15-trien-3-one (12). The structures were established by spectroscopic analyses, NMR data comparisons with similar diterpenes, and chemical correlations. All the diterpenes are assumed to have the same absolute configuration as the co-occurring (+)-stachenol (4). Diosphenol 2 and nor-lactone 5 exhibited significant potency in bioassays for cytotoxic activity against leukemia cells (L1210). Plausible biosynthetic pathways are proposed to explain the origin of the diterpene metabolites.

Diterpenes↗

PubMed2006

Comparative phytochemical characterization of three Rhodiola species.

In comparison to the well-recognized adaptogenic herb Rhodiola rosea, phytochemical constituents of two other Rhodiola species (R. heterodonta and R. semenovii) were elucidated and characterized. Two major phytochemical groups; phenolic and/or cyanogenic glycosides and proanthocyanidins, were isolated and identified in the three species. Chemical similarities among the three species were observed; however, each species displayed differences in phytochemical constituents. R. heterodonta contained a newly detected phenylethanoid glycoside, heterodontoside, in addition to the known compounds tyrosol, viridoside, salidroside, and rhodiocyanoside A. Both R. heterodonta and R. rosea contained phenylethanoid/propanoid compounds that were not detected in R. semenovii. For R. semenovii, the cyanogenic glucosides rhodiocyanoside A and lotaustralin were detected. Although the three species have proanthocyanidins composed of (-)-epigallocatechin and its 3-O-gallate esters in common, the degree of polymerization greatly differed between them. In contrast to R. heterodonta and R. semenovii, R. rosea has higher molecular weight polymeric proanthocyanidins. This study resulted in the identification and isolation of phytochemical constituents for direct cross-comparison between three Rhodiola species of medicinal and pharmacological value.

Glucosides↗

PubMed2006

Cytotoxicity of bioactive polymeric fractions from grape cell culture on human hepatocellular carcinoma, murine leukemia and non-cancerous PK15 kidney cells.

Previously, we isolated two fractions (TP-4 and TP-6) from grape cell culture that were potent catalytic inhibitors in a human DNA topoisomerase II assay for cancer chemoprevention. The objectives of this study were to further assess cytotoxicity of these fractions on cancerous and non-cancerous cells, and to subfractionate and characterize the composition of TP-6, a fraction that was selectively cytotoxic to carcinoma cell lines. Both TP-4 and TP-6 provided significant cytotoxicity to L1210 mouse leukemia cells. Only TP-6, a procyanidin-rich fraction, significantly reduced viability in HepG2 human liver cancer cells, yet unlike resveratrol, caused no cytotoxicity to non-cancerous PK15 pig kidney cells. After further subfractionation of TP-6 (maximal toxicity = 67.2%; ED(50) = 50.5 microM), the cytotoxicity of subfractions on HepG2 cells was TP-6-5 (maximal toxicity=71.8%; ED(50) = 14.1 microM), TP-6-6 (maximal toxicity=64.3%; ED(50) = 67.0 microM), and TP-6-4 (maximal toxicity = 27.6%; ED(50) = 118.0 microM) in descending order. LC-ESI/MS data suggested that cytotoxicity of these procyanidin mixtures to HepG2 cells was proportional to the degree of polymerization. Because TP-6 and its subfractions were selectively cytotoxic to cancerous cell lines tested, they warrant further investigation as potential natural anticancer agents.

Animals↗

PubMed2006

Catalytic inhibition of human DNA topoisomerase II by interactions of grape cell culture polyphenols.

Previously, we isolated mixed polyphenolic fractions on a toyopearl matrix (TP-2 to TP-6) from grape cell cultures that were highly potent catalytic inhibitors in a human DNA topoisomerase II assay for cancer chemoprevention. The objectives of this study were to evaluate the potency of, and potential interactions between, individual fractions and some of the purified bioactive polyphenols that comprise these fractions on human DNA topoisomerase II catalytic activity. Treatments that combined anthocyanin-rich fractions (TP-2; 0.5 or 2.0 microg of dried material/mL), fractions containing catechins, procyanidin dimers, and flavanones (TP-4; 0.25 microg of dried material/mL), and/or fractions enriched with procyanidin oligomers and polymers (TP-6; 0.15 or 0.5 microg of dried material/mL) showed additive effects toward catalytic inhibition of the enzyme. Epicatechin gallate (IC50 = 0.029 microM), myricetin (0.39 microM), procyanidin B2 (PB2, 4.5 microM), and resveratrol (65.7 microM), constituents of the most bioactive mixed fraction from grape cell culture (TP-4), each individually provided potent catalytic inhibition of topoisomerase II. In addition, potentiating interactions between the PB2 and the other polyphenolic constituents mentioned above and between myricetin and resveratrol were clearly demonstrated. A synergistic interaction between myricetin and resveratrol was also confirmed with isobolographic analysis at a molar ratio of 1:70.

Catalysis↗

PubMed2006

Chemopreventive activity of polyphenolics from black Jamapa bean (Phaseolus vulgaris L.) on HeLa and HaCaT cells.

The antiproliferative effects of 100% methanol crude extract and of Toyopearl and silica gel fractions from the seed coats of black Jamapa beans (Phaseolus vulgaris L.) were evaluated using HeLa, human adenocarcinoma cells, and HaCaT, human premalignant keratinocytes. The 100% methanol crude extract [172.2 microM equiv of (+)-catechin] increased adhesion of HeLa cells; however, 3- and 5-fold higher concentrations decreased the number of cells attached as a function of the treatment time. The highest concentration tested diminished the cell adhesion until 40% (after 24 h) to almost 80% (after 72 h). The IC50 values showed that the 100% methanol crude extract was the most effective inhibitor of HeLa cell proliferation, even when it was dissolved in dimethylsulfoxide (DMSO) [34.5 microM equiv of (+)-catechin] or in medium [97.7 microM equiv of (+)-catechin]. The Toyopearl 5 (TP5) fraction and silica gel 2 (SG2) fraction inhibited 60% of the HeLa cell proliferation. The IC50 was 154 microM equiv of (+)-catechin of the 100% methanol crude extract on HaCaT cells. Toyopearl fractions TP4 and TP6 significantly inhibited HaCaT cell proliferation, but the silica gel fractions did not have a significant effect. The 100% methanol crude extract (35 microg of dry material/mL) decreased the number of HeLa cells in the G0/G1 phase from 68.9% (for control cells) to 51.4% (for treated cells) and increased apoptosis (2.9 and 21.2% for control and treated cells, respectively). The results indicated that black Jamapa beans could be a source of polyphenolic compounds, which have an inhibitory effect toward HeLa cancer cells but are less aggressive on HaCaT premalignant cells.

Anticarcinogenic Agents↗

PubMed2006

Biosynthesis of 14C-phytoene from tomato cell suspension cultures (Lycopersicon esculentum) for utilization in prostate cancer cell culture studies.

This work describes the development and utilization of a plant cell culture production approach to biosynthesize and radiolabel phytoene and phytofluene for prostate cancer cell culture studies. The herbicide norflurazon was added to established cell suspension cultures of tomato (Lycopersicon esculentum cv. VFNT cherry), to induce the biosynthesis and accumulation of the lycopene precursors, phytoene and phytofluene, in their natural isomeric forms (15-cis-phytoene and two cis-phytofluene isomers). Norflurazon concentrations, solvent carrier type and concentration, and duration of culture exposure to norflurazon were screened to optimize phytoene and phytofluene synthesis. Maximum yields of both phytoene and phytofluene were achieved after 7 days of treatment with 0.03 mg norflurazon/40 mL fresh medium, provided in 0.07% solvent carrier. Introduction of 14C-sucrose to the tomato cell culture medium enabled the production of 14C-labeled phytoene for subsequent prostate tumor cell uptake studies. In DU 145 prostate tumor cells, it was determined that 15-cis-phytoene and an oxidized product of phytoene were taken up and partially metabolized by the cells. The ability to biosynthesize, radiolabel, and isolate these carotenoids from tomato cell cultures is a novel, valuable methodology for further in vitro and in vivo investigations into the roles of phytoene and phytofluene in cancer chemoprevention.

Adenocarcinoma↗

PubMed2006

Differential effects of blueberry proanthocyanidins on androgen sensitive and insensitive human prostate cancer cell lines.

Blueberries are rich in health-promoting polyphenolic compounds including proanthocyanidins. The purpose of this study was to determine if proanthocyanidin-rich fractions from both wild and cultivated blueberry fruit have the same inhibitory effects on the proliferation of LNCaP, an androgen-sensitive prostate cancer cell line, and DU145, a more aggressive androgen insensitive prostate cancer cell line. When 20 microg/ml of a wild blueberry proanthocyanidin fraction (fraction 5) was added to LNCaP media, growth was inhibited to 11% of control with an IC50 of 13.3 microg/ml. Two similar proanthocyanidin-rich fractions from cultivated blueberries (fractions 4 and 5) at the same concentration inhibited LNCaP growth to 57 and 26% of control with an IC50 of 22.7 and 5.8 microg/ml, respectively. In DU145 cells, the only fraction that significantly reduced growth compared to control was fraction 4 from cultivated blueberries with an IC50 value of 74.4 microg/ml, indicating only minor inhibitory activity. Differences in cell growth inhibition of LNCaP and DU145 cell lines by blueberry fractions rich in proanthocyanidins indicate that blueberry proanthocyanidins have an effect primarily on androgen-dependant growth of prostate cancer cells. Possible molecular mechanisms for growth inhibition are reviewed.

Androgens↗

PubMed2006

A comparative evaluation of the anticancer properties of European and American elderberry fruits.

European elderberry (Sambucus nigra), recognized in Europe for its health-promoting properties for many generations, is known to contain a range of anthocyanins, flavonoids, and other polyphenolics that contribute to the high antioxidant capacity of its berries. American elderberry (Sambucus canadensis), on the other hand, has not been cultivated, bred, and promoted as a medicinal plant like its better-characterized European counterpart. In this study, aqueous acetone extracts of the berries from these two species were fractionated and tested in a range of assays that gauge anticarcinogenic potential. Both cultivated S. nigra and wild S. canadensis fruits demonstrated significant chemopreventive potential through strong induction of quinone reductase and inhibition of cyclooxygenase-2, which is indicative of anti-initiation and antipromotion properties, respectively. In addition, fractions of S. canadensis extract showed inhibition of ornithine decarboxylase, an enzyme marker related to the promotion stage of carcinogenesis. Analysis of active fractions using mass spectrometry and liquid chromatography-mass spectrometry revealed, in addition to flavonoids, the presence of more lipophilic compounds such as sesquiterpenes, iridoid monoterpene glycosides, and phytosterols.

Anticarcinogenic Agents↗

PubMed2006

Serum testosterone is reduced following short-term phytofluene, lycopene, or tomato powder consumption in F344 rats.

Elevated serum androgens are associated with increased prostate cancer risk. Tomato consumption is also associated with reduced prostate cancer incidence, and the primary tomato carotenoid, lycopene, may modulate androgen activation in the prostate, yet little is known about other tomato carotenoids. To evaluate interrelations between phytofluene, lycopene, or tomato powder consumption and androgen status, 8-wk-old male F344 rats (fed a control AIN 93G diet) were castrated or sham-operated and subsequently provided with daily oral supplementation of phytofluene or lycopene ( approximately 0.7 mg/d) or fed a 10% tomato powder supplemented diet (AIN 93G) for 4 d. Sham-operated rats provided with either phytofluene, lycopene, or tomato powder had approximately 40-50% lower serum testosterone concentrations than the sham-operated, control-fed group. Tissue and serum phytofluene and lycopene concentrations were greater in castrated rats than in sham-operated rats, which may have been due in part to a decrease of hepatic CYP 3A1 mRNA expression and benzyloxyresorufin-O-dealkylase activity. Some changes in prostatic and testicular steroidogenic enzyme mRNA expression were found; in particular, prostate 17 beta-hydroxysteroid dehydrogenase 4 mRNA expression in castrated rats fed lycopene or tomato powder was 1.7-fold that of the sham-operated, control-fed group. Modest changes in mRNA expression of steroidogenic enzymes with short-term carotenoid intake may alter the flux of androgen synthesis to less potent compounds. Overall, results illustrate that short-term intake of tomato carotenoids significantly alters androgen status, which may partially be a mechanism by which tomato intake reduces prostate cancer risk.

17-Hydroxysteroid Dehydrogenases↗

PubMed2005

Characterization of polyphenolics in the seed coat of Black Jamapa bean (Phaseolus vulgaris L.).

The common bean contains phytochemicals, including phenolic compounds, which can provide health benefits to the consumer. Our objective was to characterize the polyphenolic compounds present in the seed coat of Black Jamapa bean and to test fractionation methods that permit the recovery of polyphenolics in their naturally occurring forms. A 100% methanol extract from seed coats was subjected to different chromatographic fractionation methods. Analysis by HPLC-MS revealed that a better separation of phytochemicals was achieved using direct silica gel fractionation, which allowed more accurate identification of compounds, especially of the flavonols. Anthocyanins, flavanol monomers, and heterogeneous flavanol oligomers up to hexamers were detected. To our knowledge, this is the first time that myricetin glycoside and proanthocyanidin oligomers containing (epi)-gallocatechin have been reported in the black bean. The fractionation methods used in this study produced large quantities of natural mixtures of flavonoids suitable for testing bioactivity and phytochemical interactions.

Anthocyanins↗

PubMed2005

Effects of grape cell culture extracts on human topoisomerase II catalytic activity and characterization of active fractions.

Grape and its cell culture extracts are rich in flavonoids and stilbenes that are biologically active. The objective of this study was to evaluate possible inhibitory effects of grape (a Vitis hybrid Bailey Alicant A) cell culture extract and subfractions on human DNA topoisomerase II catalytic activity and to characterize constituents in the most potent fractions. At 5 microg/mL, grape cell crude extract and Toyopearl (TP) fractions 2-6 provided significantly greater inhibition of topoisomerase II catalytic activity than quercetin, a chemopreventive agent previously known as a topoisomerase catalytic inhibitor. The most potent topoisomerase II catalytic inhibitors from grape cell culture extracts in descending order of potency were TP fractions 4 and 6 (IC(50) = 0.28-0.29 microg/mL), TP-3 (IC(50) = 0.74 microg/mL), and crude extract (IC(50) = 1.02 microg/mL); each was significantly more potent than resveratrol (IC(50) = 18.0 microg/mL), another well-known chemopreventive topoisomerase II catalytic inhibitor. Using both high-performance liquid chromatography and liquid chromatography electrospray ionization mass spectrometry, constituents in TP-4 and TP-6 were characterized. These constituents included cyanidin-3,5-diglucoside, malvidin-3-acetylglucoside, peonidin-3-coumaryl-5-diglucoside, procyanidin B(1), procyanidin B(2), procyanidin B(5), procyanidin dimer digallate, procyanidin C(1), myricetin, and rutin, none of which have been previously characterized from grape cell cultures. The significant potency especially of TP-4 and TP-6 from grape cell cultures suggests that these fractions may have potential as chemopreventive agents.

Anticarcinogenic Agents↗

PubMed2005

Sorting out bioactivity in flavonoid mixtures.

Natural polyphenolic compounds, including flavonoids, which are synthesized by plants, have repeatedly been linked to multiple health benefits, but specific bioactive compounds or individualized roles have been difficult to interpret or to assign. This limitation has prevented the development of robust dietary recommendations. This paper describes a plant-cell-culture production approach that provides uniform yields of abundant natural mixtures of flavonoids from small molecules to high molecular weight oligomers and polymers. The flavonoids correspond to compounds produced in nature by the same plant genotypes; but, through elicitation tactics, the levels and the profiles of the flavonoid yield can be enhanced, and isolation from cell cultures is more rapid and streamlined than isolation from in vivo plant tissues. Introduction of (14)C sucrose to the cell-culture medium enabled the recovery and the identification of biolabeled flavonoid mixtures, which could be easily tracked in serum of rats after gavage. The biolabeled materials are now being investigated to determine tissue localization, structures of bioactive components, and the interactions between flavonoid components that contribute to bioactive potential.

Animals↗

PubMed2004

Effective separation of potent antiproliferation and antiadhesion components from wild blueberry (Vaccinium angustifolium Ait.) fruits.

Extracts from wild blueberry (Vaccinium angustifolium Ait.) were separated into proanthocyanidin-rich fractions using liquid vacuum and open column chromatography on Toyopearl and Sephadex LH-20, respectively. Fractions were characterized using analytical tools including mass spectrometry and NMR spectroscopy; fraction composition was correlated with bioactivity using antiproliferation and antiadhesion in vitro assays. There was a significant positive correlation between proanthocyanidin content of different fractions and biological activity in both the antiproliferation and antiadhesion assays. Two fractions containing primarily 4-->8-linked oligomeric proanthocyanidins with average degrees of polymerization (DPn) of 3.25 and 5.65 inhibited adhesion of Escherichia coli responsible for urinary tract infections. Only the fraction with a DPn of 5.65 had significant antiproliferation activity against human prostate and mouse liver cancer cell lines. These findings suggest both antiadhesion and antiproliferation activity are associated with high molecular weight proanthocyanidin oligomers found in wild blueberry fruits.

Animals↗

PubMed2004

Biosynthesis and characterization of 14C-enriched flavonoid fractions from plant cell suspension cultures.

A range of radiolabeled anthocyanins, proanthocyanidins, and other flavonoids were accumulated by cell suspension cultures of two plant species, ohelo (Vaccinium pahalae) and grape (a Vitis hybrid, Bailey Alicant A), after providing uniformly labeled [(14)C]sucrose to the medium. Approximately 15% of administered label was recovered in a series of flavonoid-rich fractions varying in composition. Anthocyanins, and monomers to oligomers of proanthocyanidins, were labeled effectively and characterized from both species. Most of the proanthocyanidin oligomers were based on the flavan-3-ols (+)-catechin and (-)-epicatechin. Cyanidin and peonidin glycosides were the dominant forms of anthocyanins in both species. Whereas the predominant form of flavonoids identified from ohelo cell cultures was proanthocyanidins, grape cell cultures produced mostly anthocyanins. The labeled phytochemicals were produced for use in subsequent in vivo animal feeding studies to gauge their bioavailability and accumulation in target organs.

Anthocyanins↗

PubMed2004

Anthocyanins and Human Health: An In Vitro Investigative Approach.

Anthocyanin pigments and associated flavonoids have demonstrated ability to protect against a myriad of human diseases, yet they have been notoriously difficult to study with regard to human health. Anthocyanins frequently interact with other phytochemicals to potentiate biological effects, thus contributions from individual components are difficult to decipher. The complex, multicomponent structure of compounds in a bioactive mixture and the degradation of flavonoids during harsh extraction procedures obscure the precise assignment of bioactivity to individual pigments. Extensive metabolic breakdown after ingestion complicates tracking of anthocyanins to assess absorption, bioavailability, and accumulation in various organs. Anthocyanin pigments and other flavonoids that are uniformly, predictably produced in rigorously controlled plant cell culture systems can be a great advantage for health and nutrition research because they are quickly, easily isolated, lack interferences found in whole fruits, can be elicited to provoke rapid and prolific accumulation, and are amenable to biolabeling so that metabolic fate can be investigated after ingestion.

Journal Article↗

PubMed2003

[14C]-lycopene and [14C]-labeled polar products are differentially distributed in tissues of F344 rats prefed lycopene.

Epidemiologic evidence suggests a possible role for lycopene-rich foods in the prevention of prostate cancer and cardiovascular disease. Despite active research in disease reduction, there is a paucity of information on the absorption, biodistribution and metabolism of lycopene. The aim of this study was to evaluate the biodistribution of 14C-lycopene (specific activity, 1.83 microCi/mg) and 14C-labeled products after an oral dose of 22 microCi of 14C-lycopene in male rats that had been prefed a lycopene-containing diet (0.25 g lycopene/ kg diet) for 30 d. The percentage of 14C excreted in feces and urine over the 168 h was 68%. Quantitatively, serum 14C levels were maintained between 3 and 24 h then decreased at 72 h (P < 0.05). At all time points the majority of tissue 14C was in the liver (approximately 72%), although total hepatic 14C decreased after 24 h. In a comparison of the extrahepatic tissue at 168 h, the 14C was greatest in adipose tissue followed by spleen and then adrenal; approximately 80% of the 14C in the liver was in the cis and all-trans configuration at all time points. At 3 h, the 14C in seminal vesicles was primarily in the all-trans plus 5-cis forms (70%), but by 168 h, 55% of 14C was present as 14C-polar products. Despite the presence of unlabeled lycopene in the prostate, the primary 14C form was in 14C-polar products (67-92%), even at 3 h. The percentage and amount of 14C-polar products in the dorsolateral prostate lobe increased from 3 to 24 h and then reached a plateau. The data suggest that lycopene may be metabolized differently among tissues in rats prefed lycopene.

Adrenal Glands↗