Withanolides and Sucrose Esters from Physalis neomexicana.

Four withanolides (1-4) and two sucrose esters (5, 6) were isolated from the aerial parts of Physalis neomexicana. The structures of 1-6 were elucidated through a variety of spectroscopic techniques. Cytotoxicity studies of the isolates revealed that 2 inhibited human breast cancer cell lines (MDA-MB-231 and MCF-7) with IC50 values of 1.7 and 6.3 µM, respectively.

Ethnic differences in medicinal plant use among University students: a cross-sectional survey of self-reported medicinal plant use at two Midwest Universities.

BACKGROUND: Numerous surveys of medicinal plant use among college students abound, but none compare use between students enrolled in two different Universities with significantly different ethnic compositions. The objective of this study is to compare medicinal plant use between two different ethnic college populations and explore differences between student medicinal plant users and non-users for comparison with previous research. METHODS: Students (n = 721) at a large research university (n = 498) and a Pan-Tribal University for Native Americans (n = 233) completed surveys in October 2011 to assess past year medicinal plant use. The Mann-Whitney U test, Chi Square test, and General Linear Model were used to compare demographics and self-reported use of medicinal plants among students at both Universities and between past year users and non-users. RESULTS: Over 23% of university students surveyed reported past year medicinal plant use. Users were more likely to use commercial tobacco products and to report poorer health than non-users. While Native American student medicinal plant users reported significantly higher rates of commercial tobacco use, lower self-assessment of health, and less use of prescription medicine than non-Native users, no significant differences in prevalence of medicinal plant use were found between University student populations. CONCLUSIONS: Results are consistent with preexisting data showing higher rates of medicinal plant use among college students compared to the larger US population of adults and demonstrate previously documented health disparities in Native American populations compared to non-Native Americans.

Withanolides from Physalis hispida.

Nine new withanolides (1-9), withahisolides A-I, were isolated along with nine known compounds (10-18) from the aerial parts of Physalis hispida. The structures of 1-9 were elucidated through a variety of spectroscopic techniques, while the structures of 1 and 2 were confirmed by X-ray crystallographic analysis. Compounds 1-3 are the first withanolides with nonaromatic six-membered ring D moieties. In addition, withanolide 8 represents a novel withanolide skeleton due to the absence of a C-13-C-17 bond within the steroidal nucleus.

Antiproliferative withanolides from Datura wrightii.

A new withanolide, named withawrightolide (1), and four known withanolides (2-5) were isolated from the aerial parts of Datura wrightii. The structure of compound 1 was elucidated through 2D NMR and other spectroscopic techniques. In addition, the structure of withametelin L (2) was confirmed by X-ray crystallographic analysis. Using MTS viability assays, withanolides 1-5 showed antiproliferative activities against human glioblastoma (U251 and U87), head and neck squamous cell carcinoma (MDA-1986), and normal fetal lung fibroblast (MRC-5) cells with IC50 values in the range between 0.56 and 5.6 µM.

Cytotoxic cardiac glycosides and other compounds from Asclepias syriaca.

Phytochemical investigation of the dried biomass of Asclepias syriaca afforded five new compounds (1-5), along with 19 known structures. Overall, the secondary metabolites isolated and identified from this plant showed a wide structural diversity including pentacyclic triterpenes, cardiac glycosides, flavonoid glycosides, lignans, a phenylethanoid, and a glycosylated megastigmane. In addition, the isolates were tested against the cancer breast cell line Hs578T, and those showing IC(50) values lower than 50 µM (1 and 6-9) were further investigated in three additional breast cancer cell lines (MCF-7, T47D, and Sk-Br-3) and the normal breast cell line Hs578Bst.

Benzyl benzoate glycosides from Oligoneuron rigidum.

Two new benzyl benzoate glycosides were isolated from the leaves and stems of the native North American prairie plant Oligoneuron rigidum (stiff goldenrod). The glycosides were isolated as a mixture of benzoate and acetate esters, which were subjected to mild base hydrolysis to facilitate full structural characterization using LCMSMS and 1D and 2D NMR data.

Minor withanolides of Physalis longifolia: structure and cytotoxicity.

In our recent publication on bioactive guided isolation of compounds from Physalis longifolia (Solanaceae) novel anti-proliferative agents withalongolides A (4) and B (5), and their highly cytotoxic analogues, withalongolide A 4,19,27-triacetate (4a) and withalongolide B 4,19-diacetate (5a) were elucidated. In this study, the two lead compounds (4, 5) were re-isolated in gram quantities for the purpose of further analogue preparation and in vivo testing that would continue to probe structure-activity relationships. During this process, two additional withanolides, named withalongolides O (1) and P (2), were elucidated. Their structures were determined by spectroscopic techniques with 1 being subsequently confirmed by X-ray crystallographic analysis. Utilizing a MTS [3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium] viability assay, withalongolide O (1) and its 4,7-diaceatate (1a), both containing the functionalities of Δ(2)-1-oxo- in A ring, a 5ß,6ß-epoxy in B ring, and a lactone ring in the nine-carbon side chain, exhibited potent cytotoxicity against human head and neck squamous cell carcinoma (JMAR and MDA-1986), melanoma (B16F10 and SKMEL-28), and normal fetal lung fibroblast (MRC-5) cells with IC(50) values in the range between 0.15 and 2.95 µM. In addition, the previously reported α orientation of 7-acetate group in acnistins C and D should be revised to the ß orientation on the basis of NMR data comparison.

Cytotoxic withanolide constituents of Physalis longifolia.

Fourteen new withanolides, 1-14, named withalongolides A-N, respectively, were isolated from the aerial parts of Physalis longifolia together with eight known compounds (15-22). The structures of compounds 1-14 were elucidated through spectroscopic techniques and chemical methods. In addition, the structures of withanolides 1, 2, 3, and 6 were confirmed by X-ray crystallographic analysis. Using a MTS viability assay, eight withanolides (1, 2, 3, 7, 8, 15, 16, and 19) and four acetylated derivatives (1a, 1b, 2a, and 2b) showed potent cytotoxicity against human head and neck squamous cell carcinoma (JMAR and MDA-1986), melanoma (B16F10 and SKMEL-28), and normal fetal fibroblast (MRC-5) cells with IC50 values in the range between 0.067 and 9.3 µM.

Montane meadow change during drought varies with background hydrologic regime and plant functional group.

Climate change models for many ecosystems predict more extreme climatic events in the future, including exacerbated drought conditions. Here we assess the effects of drought by quantifying temporal variation in community composition of a complex montane meadow landscape characterized by a hydrological gradient. The meadows occur in two regions of the Greater Yellowstone Ecosystem (Gallatin and Teton) and were classified into six categories (M1-M6, designating hydric to xeric) based upon Satellite pour l'Observation de la Terre (SPOT) satellite imagery. Both regions have similar plant communities, but patch sizes of meadows are much smaller in the Gallatin region. We measured changes in the percent cover of bare ground and plants by species and functional groups during five years between 1997 and 2007. We hypothesized that drought effects would not be manifested evenly across the hydrological gradient, but rather would be observed as hotspots of change in some areas and minimally evident in others. We also expected varying responses by plant functional groups (forbs vs. woody plants). Forbs, which typically use water from relatively shallow s,oils compared to woody plants, were expected to decrease in cover in mesic meadows, but increase in hydric meadows. Woody plants, such as Artemisia, were expected to increase, especially in mesic meadows. We identified several important trends in our meadow plant communities during this period of drought: (1) bare ground increased significantly in xeric meadows of both regions (Gallatin M6 and Teton M5) and in mesic (M3) meadows of the Teton, (2) forbs decreased significantly in the mesic and xeric meadows in both regions, (3) forbs increased in hydric (M1) meadows of the Gallatin region, and (4) woody species showed increases in M2 and M5 meadows of the Teton region and in M3 meadows of the Gallatin region. The woody response was dominated by changes in Artemisia spp. and Chrysothamnus viscidiflorus. Thus, our results supported our expectations that community change was not uniform across the landscape, but instead could be predicted based upon functional group responses to the spatial and temporal patterns of water availability, which are largely a function of plant water use and the hydrological gradient.

Effects of hay management and native species sowing on grassland community structure, biomass, and restoration.

Prairie hay meadows are important reservoirs of grassland biodiversity in the tallgrass prairie regions of the central United States and are the object of increasing attention for conservation and restoration. In addition, there is growing interest in the potential use of such low-input, high-diversity (LIHD) native grasslands for biofuel production. The uplands of eastern Kansas, USA, which prior to European settlement were dominated by tallgrass prairie, are currently utilized for intensive agriculture or exist in a state of abandonment from agriculture. The dominant grasslands in the region are currently high-input, low-diversity (HILD) hay fields seeded to introduced C3 hay grasses. We present results from a long-term experiment conducted in a recently abandoned HILD hay field in eastern Kansas to evaluate effects of fertilization, haying, and native species sowing on community dynamics, biomass, and potential for restoration to native LIHD hay meadow. Fertilized plots maintained dominance by introduced grasses, maintained low diversity, and were largely resistant to colonization throughout the study. Non-fertilized plots exhibited rapid successional turnover, increased diversity, and increased abundance of C4 grasses over time. Haying led to modest changes in species composition and lessened the negative impact of fertilization on diversity. In non-fertilized plots, sowing increased representation by native species and increased diversity, successional turnover, and biomass production. Our results support the shifting limitations hypothesis of community organization and highlight the importance of species pools and seed limitations in constraining successional turnover, community structure, and ecosystem productivity under conditions of low fertility. Our findings also indicate that several biological and functional aspects of LIHD hay meadows can be restored from abandoned HILD hay fields by ceasing fertilization and reintroducing native species through sowing. Declines in primary production and hay yield that result from the cessation of fertilization may be at least partially compensated for by restoration.

Evidence for the Paleoethnobotany of the Neanderthal: A Review of the Literature.

Our perception of our closest human relatives, the Neanderthals, has evolved in the last few decades from brutish ape-men to intelligent archaic human peoples. Our understanding and appreciation of their cultural sophistication has only recently extended to their diet. Only within the last few years, with new techniques and a shift in focus, have we begun to truly investigate and understand the role of plants in their diet and culture. The more we learn about Neanderthals, the more we realize that biological and cultural distinctions between them and us were relatively small. Given that we coexisted and likely interacted with them for thousands of years, the more we learn about them, the better we may understand our own past. In that light, we review the current evidence, derived from such sources as plant remains (e.g., starch, pollen, phytoliths, and seeds) in soil and dental calculus, dental and tool wear, coprolites, and genetics, for Neanderthal's nutritional, medicinal, and ritual use of plants, which includes 61 different taxa from 26 different plant families found at 17 different archaeological sites. Further, we updated and standardized botanical nomenclature from many sources published over many decades to provide a more stable foundation for future work.

Withanolides from Physalis coztomatl.

Six withanolides (1-6), as well as two known withanolides (physachenolide D 7 and withanoside VI 8), were isolated from the aerial parts of Physalis coztomatl (Solanaceae). Structural elucidations of 1-6 were achieved through 2D NMR and other spectroscopic techniques, while the structure of 1 was confirmed by X-ray crystallographic analysis. In addition, the stereochemical orientation of the 17-hydroxy group in withanolides was discussed in relation to (13)C NMR shifts of C-12, 13, 14 and 16. Such analysis established that coagulansin A contains a 17α-hydroxy moiety rather than the reported 17ß-hydroxy functionality, and has been revised accordingly.

Analysis of Major Withanolides in Physalis longifolia Nutt. by HPLC-PDA.

An analytical method based on high-performance liquid chromatography-photodiode array detection was developed for the simultaneous determination of three anti-proliferative withanolides [withalongolide A ( 1: ), withaferin A ( 2: ) and withalongolide B ( 3: )] present in the aboveground biomass of the long-leaf groundcherry, Physalis longifolia. This method was achieved by biomass extraction followed by chromatographic separation on C18 column eluted with a gradient acetonitrile-water mobile phase. Calibration curves produced satisfactory linear regression (r(2) > 0.9995) for each examined sample. The method was also validated for accuracy, precision and limits of detection and quantification. Such an approach is applicable for the rapid detection and quantitative assessment of withanolides in various P. longifolia accessions.

Verticillosides A-M: Polyoxygenated pregnane glycosides from Asclepias verticillata L.

As part of our ongoing effort to explore the chemical diversity of plants of the United States Midwest region, the isolation and identification of 13 pregnane glycosides named verticillosides A-M from Asclepias verticillata L. are reported. The structures of these compounds were elucidated by various spectroscopic techniques, including 1D and 2D NMR, IR, UV, and HRMS. The cytotoxicity of the isolates was evaluated against paired breast cell lines Hs578T (cancer) and Hs578Bst (normal), however, no significant growth inhibition was observed.

Novel withanolides target medullary thyroid cancer through inhibition of both RET phosphorylation and the mammalian target of rapamycin pathway.

BACKGROUND: Despite development of current targeted therapies for medullary thyroid cancer (MTC), long-term survival remains unchanged. Recently isolated novel withanolide compounds from Solanaceae physalis are highly potent against MTCs. We hypothesize that these withanolides uniquely inhibit RET phosphorylation and the mammalian target of rapamycin (mTOR) pathway in MTC cells as a mechanism of antiproliferation and apoptosis. METHODS: MTC cells were treated with novel withanolides and MTC-targeted drugs. In vitro studies assessed cell viability and proliferation (MTS; trypan blue assays), apoptosis (flow cytometry with Annexin V/PI staining; confirmed by Western blot analysis), long-term cytotoxic effects (clonogenic assay), and suppression of key regulatory proteins such as RET, Akt, and mTOR (by Western blot analysis). RESULTS: The novel withanolides potently reduced MTC cell viability (half maximal inhibitory concentration [IC(50)], 270-2,850 nmol/L; 250-1,380 nmol/L for vandetanib; 360-1,640 nmol/L for cabozantinib) with induction of apoptosis at <1,000 nmol/L of drug. Unique from other targeted therapies, withanolides suppressed RET and Akt phosphorylation and protein expression (in a concentration- and time-dependent manner) as well as mTOR activity and translational activity of 4E-BP1 and protein synthesis mediated by p70S6kinase activation at IC(50) concentrations. CONCLUSION: Novel withanolides from Physalis selectively and potently inhibit MTC cells in vitro. Unlike other MTC-targeted therapies, these compounds uniquely inhibit both RET kinase activity and the Akt/mTOR prosurvival pathway. Further translational studies are warranted to evaluate their clinical potential.

Characterization of an anti-tuberculosis resin glycoside from the prairie medicinal plant Ipomoea leptophylla.

The organic soluble extract from the leaves of the native North American prairie plant Ipomoea leptophylla (big root morning glory) showed in vitro activity against M. tuberculosis. Bioassay-guided fractionation of this extract resulted in the identification of two new resin glycosides (6, 7). Base-catalyzed hydrolysis of these glycosides gave operculinic acid (1) as the glycosidic acid component as well as trans-cinnamic acid, propanoic acid, and lauric acid. The complete structure elucidation was accomplished through derivatization, 1D and 2D NMR spectroscopy (TOCSY, ROESY, HSQC, HMBC), and MS/MS experiments on 6 and 7 as well as the permethylated derivative 8.