Activation of heat shock and antioxidant responses by the natural product celastrol: transcriptional signatures of a thiol-targeted molecule.

Stress response pathways allow cells to sense and respond to environmental changes and adverse pathophysiological states. Pharmacological modulation of cellular stress pathways has implications in the treatment of human diseases, including neurodegenerative disorders, cardiovascular disease, and cancer. The quinone methide triterpene celastrol, derived from a traditional Chinese medicinal herb, has numerous pharmacological properties, and it is a potent activator of the mammalian heat shock transcription factor HSF1. However, its mode of action and spectrum of cellular targets are poorly understood. We show here that celastrol activates Hsf1 in Saccharomyces cerevisiae at a similar effective concentration seen in mammalian cells. Transcriptional profiling revealed that celastrol treatment induces a battery of oxidant defense genes in addition to heat shock genes. Celastrol activated the yeast Yap1 oxidant defense transcription factor via the carboxy-terminal redox center that responds to electrophilic compounds. Antioxidant response genes were likewise induced in mammalian cells, demonstrating that the activation of two major cell stress pathways by celastrol is conserved. We report that celastrol's biological effects, including inhibition of glucocorticoid receptor activity, can be blocked by the addition of excess free thiol, suggesting a chemical mechanism for biological activity based on modification of key reactive thiols by this natural product.

Role of Ashwangandha (Withania Somnifera) in Immune Modulation: Proposed Influence in Immune-Regulation

The immune system evolved to protect organisms from an infinite variety of disease-causing agents but to avoid harmful responses to self. However, such a powerf~dl efense mechanism requires regulation. Immune regulation includes homeostatic and cellmediated targeted mechanisms to the activation, differentiation and function of antigen-triggered immuno-competent cells and irnmunoregulatory cells. The regulation of the immune system has been a major challenge for the management of autoimmune disorders, tumor immunity, infectious diseases and organ transplants. However, irnmuno-modulatory procedures used by modern medicine to induce immunoregulatory function have deleterious side effects. Ashwangandha (Withania somnifera), an herb used in Ayurvedic medicine is being tested and used in experimental and clinical cases with potential immuno-modulatory functions without any side effects. Here we propose future usages of Ashwangandha for immuno-regulatory function in translational research.

Evidence from the real world: N-15 natural abundances reveal enhanced nitrogen use at high plant diversity in Central European grasslands

Complementarity that leads to more efficient resource use is presumed to be a key mechanism explaining positive biodiversity–productivity relationships but has been described solely for experimental set-ups with controlled environmental settings or for very short gradients of abiotic conditions, land-use intensity and biodiversity. Therefore, we analysed plant diversity effects on nitrogen dynamics across a broad range of Central European grasslands. The 15N natural abundance in soil and plant biomass reflects the net effect of processes affecting ecosystem N dynamics. This includes the mechanism of complementary resource utilization that causes a decrease in the 15N isotopic signal. We measured plant species richness, natural abundance of 15N in soil and plants, above-ground biomass of the community and three single species (an herb, grass and legume) and a variety of additional environmental variables in 150 grassland plots in three regions of Germany. To explore the drivers of the nitrogen dynamics, we performed several analyses of covariance treating the 15N isotopic signals as a function of plant diversity and a large set of covariates. Increasing plant diversity was consistently linked to decreased δ15N isotopic signals in soil, above-ground community biomass and the three single species. Even after accounting for multiple covariates, plant diversity remained the strongest predictor of δ15N isotopic signals suggesting that higher plant diversity leads to a more closed nitrogen cycle due to more efficient nitrogen use. Factors linked to increased δ15N values included the amount of nitrogen taken up, soil moisture and land-use intensity (particularly fertilization), all indicators of the openness of the nitrogen cycle due to enhanced N-turnover and subsequent losses. Study region was significantly related to the δ15N isotopic signals indicating that regional peculiarities such as former intensive land use could strongly affect nitrogen dynamics. Synthesis. Our results provide strong evidence that the mechanism of complementary resource utilization operates in real-world grasslands where multiple external factors affect nitrogen dynamics. Although single species may differ in effect size, actively increasing total plant diversity in grasslands could be an option to more effectively use nitrogen resources and to reduce the negative environmental impacts of nitrogen losses.

Lateglacial and Holocene vegetation belts in the Pirin Mountains (southwestern Bulgaria)

Pollen stratigraphy of a core 270 cm long from Lake Dalgoto at 2310 m in the Northern Pirin Mountains, southern Bulgaria, was treated by optimal partitioning and compared to a broken-stick model to reveal statistically significant pollen zones. The vegetational reconstructions presented here are based on pollen percentages and pollen influx, on comparisons of modern and fossil pollen spectra, and on macrofossil dates from other sites in the mountains. During the Younger Dryas (11000–10200 14C yr BP), an open xerophytic herb vegetation with Artemisia and Chenopodiaceae was widely developed around the lake. Deciduous trees growing at lower elevations contributed to the pollen rain deposited at the higher-elevation sampling sites. Specifically, from 10200 to 8500 yr BP, Quercus, Ulmus, Tilia and Betula expanded rapidly at low and intermediate elevations, and between 8500 and 6500 yr BP they extended to higher elevations close to the upper forest limit, which was formed by Betula pendula at about 1900 m. Coniferous species were limited in the region at this time. After 6500 yr BP, the expansion of conifers (Pinus peuce, P. sylvestris, P. mugo, Abies alba) at high elevations forced the deciduous trees downward. Between 6500 and 3000 yr BP, the forest limit at 2200 m was formed by P. peuce, and A. alba had its maximum vertical range up to 1900 m. Later the abundance and vertical range of P. peuce and A. alba were reduced. After 3000 yr BP, Picea expanded.

Drivers of nitrogen leaching from organic layers in Central European beech forests

Background and Aims: The response of forest ecosystems to continuous nitrogen (N) deposition is still uncertain. We investigated imports and exports of dissolved N from mull-type organic layers to identify the controls of N leaching in Central European beech forests under continuous N deposition. Methods: Dissolved N fluxes with throughfall and through mull-type organic layers (litter leachate) were measured continuously in 12 beech forests on calcareous soil in two regions in Germany over three consecutive growing seasons. Results Mean growing season net (i.e. litter leachate – throughfall flux) fluxes of total dissolved N (TDN) from the organic layer were low (2.3 ± 5.6 kg ha −1 ) but varied widely from 12.9 kg ha −1 to –8.3 kg ha −1 . The small increase of dissolved N fluxes during the water passage through mull-type organic layers suggested that high turnover rates coincided with high microbial N assimilation and plant N uptake. Stand basal area had a positive feedback on N fluxes by providing litter for soil organic matter forma- tion. Plant diversity, especially herb diversity, reduced dissolved N fluxes. Soil fauna biomass increased NO3−-N fluxes with litter leachate by stimulating mineralization. Microbial biomass measures were not related to dissolved N fluxes. Conclusions Our results show that dissolved N exports from organic layers contain significant amounts of throughfall-derived N (mainly NO3−-N) that flushes through the organic layer but also highlight that N leaching from organic layers is driven by the complex interplay of plants, animals and microbes. Furthermore, diverse understories reduce N leaching from Central European beech forests.

Modulatory Effects of Eschscholzia californica Alkaloids on Recombinant GABAA Receptors.

The California poppy (Eschscholzia californica Cham.) contains a variety of natural compounds including several alkaloids found exclusively in this plant. Because of the sedative, anxiolytic, and analgesic effects, this herb is currently sold in pharmacies in many countries. However, our understanding of these biological effects at the molecular level is still lacking. Alkaloids detected in E. californica could be hypothesized to act at GABAA receptors, which are widely expressed in the brain mainly at the inhibitory interneurons. Electrophysiological studies on a recombinant α 1 β 2 γ 2 GABAA receptor showed no effect of N-methyllaurotetanine at concentrations lower than 30 μM. However, (S)-reticuline behaved as positive allosteric modulator at the α 3, α 5, and α 6 isoforms of GABAA receptors. The depressant properties of aerial parts of E. californica are assigned to chloride-current modulation by (S)-reticuline at the α 3 β 2 γ 2 and α 5 β 2 γ 2 GABAA receptors. Interestingly, α 1, α 3, and α 5 were not significantly affected by (R)-reticuline, 1,2-tetrahydroreticuline, codeine, and morphine-suspected (S)-reticuline metabolites in the rodent brain.

Transgenerational effects of land use on offspring performance and growth in Trifolium repens

entral European grasslands vary widely in productivity and in mowing and grazing regimes. The resulting differences in competition and heterogeneity among grasslands might have direct effects on plants, but might also affect the growth and morphology of their offspring through maternal effects or adaptive evolution. To test for such transgenerational effects, we grew plants of the clonal herb Trifolium repens from seeds collected in 58 grassland sites differing in productivity and mowing and grazing intensities in different treatments: without competition, with homogeneous competition, and with heterogeneous competition. In the competition-free treatment, T. repens from more productive, less frequently mown, and less intensively grazed sites produced more vegetative offspring, but this was not the case in the other treatments. When grown among or in close proximity to competitors, T. repens plants did not show preferential growth towards open spaces (i.e., no horizontal foraging), but did show strong vertical foraging by petiole elongation. In the homogeneous competition treatment, petiole length increased with the productivity of the parental site, but this was not the case in the heterogeneous competition treatment. Moreover, petiole length increased with mowing frequency and grazing intensity of the parental site in all but the homogeneous competition treatment. In summary, although the expression of differences between plants from sites with different productivities and land-use intensities depended on the experimental treatment, our findings imply that there are transgenerational effects of land use on the morphology and performance of T. repens.

Plant diversity moderates drought stress in grasslands: Implications from a large real-world study on 13C natural abundances

Land-use change and intensification play a key role in the current biodiversity crisis. The resulting species loss can have severe effects on ecosystem functions and services, thereby increasing ecosystem vulnerability to climate change. We explored whether land-use intensification (i.e. fertilization intensity), plant diversity and other potentially confounding environmental factors may be significantly related to water use (i.e. drought stress) of grassland plants. Drought stress was assessed using δ13C abundances in aboveground plant biomass of 150 grassland plots across a gradient of land-use intensity. Under water shortage, plants are forced to increasingly take up the heavier 13C due to closing stomata leading to an enrichment of 13C in biomass. Plants were sampled at the community level and for single species, which belong to three different functional groups (one grass, one herb, two legumes). Results show that plant diversity was significantly related to the δ13C signal in community, grass and legume biomass indicating that drought stress was lower under higher diversity, although this relation was not significant for the herb species under study. Fertilization, in turn, mostly increased drought stress as indicated by more positive δ13C values. This effect was mostly indirect by decreasing plant diversity. In line with these results, we found similar patterns in the δ13C signal of the organic matter in the topsoil, indicating a long history of these processes. Our study provided strong indication for a positive biodiversity-ecosystem functioning relationship with reduced drought stress at higher plant diversity. However, it also underlined a negative reinforcing situation: as land-use intensification decreases plant diversity in grasslands, this might subsequently increases drought sensitivity. Vice-versa, enhancing plant diversity in species-poor agricultural grasslands may moderate negative effects of future climate change.

An ethnographic study of home remedy use for African-American children

Background and significance. The use of herbs and other remedies by adult and elderly African-Americans has been documented. However, little is understood regarding the use of herbs for African-American children. The purpose of this study was to document and describe the historical and present day uses of herbal and other remedies, specifically for the health and illness of African-American children. This information will provide health care providers with a better understanding of their African-American patients. This information may also contribute to the emerging appreciation of indigenous uses of phytotherapeutics. ^ Methods. A focused ethnographic approach was used to describe the cultural context, including the beliefs, values, customs, and behaviors of a particular culture. The use of intensive fieldwork, including participant observation, audiotaped formal interviews, photographs, and specimen collection of plants helped to describe herb use in this population. Information on the growing, harvesting, preparation, and storage of these plant remedies, as well as the amount and dosage of these compounds was collected in a typology. Detailed information was gathered to discern how, when, and under what conditions these remedies were used and their expected results. Further data collection focused on the history of herbal use, and explanations for how and why informants thought the herbs work. ^ Setting and participants. The setting for this study was in East Texas and field work extended over the period of one year. Thirty African-Americans, age 38 to 98, were interviewed for the study. The African-American population in this area has been relatively stable, with roots dating back prior to the reconstruction period, which allowed excellent historical information. Informants were chosen by a nominated sampling technique starting with two key informants knowledgeable about the use of home remedies for children. ^ Findings. The findings of this study suggest that African-American children in East Texas have a long history of receiving herbs and home remedies for health promotion and illness. Data further suggests that there is a strong connection between spirituality and the health beliefs and practices of this community. This spiritual component underlies the accuracy of oral recall for remedies that have been used over many generations and the use of natural folk remedies. A typology of the herbal remedies was developed with folk and Latin name, herb place of origin, known scientific properties, and informant folk usage and dosage information. ^

Evidence that lymphokine -activated killer (LAK) cell function is regulated by both serine and tyrosine kinase pathways

Signal transduction pathways operative in lymphokine activated killer (LAK) cells during execution of cytolytic function have never been characterized. Based on ubiquitous involvement of protein phosphorylation in activation of cytolytic mechanisms used by CTL and NK cells, it was hypothesized that changes in protein phosphorylation should occur when LAK encounter tumor targets. It was further hypothesized that protein kinases would regulate LAK-mediated cytotoxicity. Exposure to either SK-Mel-1 (melanoma) or Raji (lymphoma) targets consistently led to increased phosphorylation of two 65-kD LAK proteins pp65a and -b, with isoelectric points (pI) of 5.1 and 5.2 respectively. Increased p65 phosphorylation was initiated between 1 and 5 min after tumor coincubation, occurred on Ser residues, required physical contact between LAK and tumors, correlated with target recognition, and also occurred after crosslinking Fc$\gamma$RIIIA in the absence of tumors. Both pp65a and -b were tentatively identified as phosphorylated forms of the actin-bundling protein L-plastin, based on pI, molecular weight, and cross-reactivity with specific antiserum. The known biochemical properties of L-plastin suggest it may be involved in regulating adhesion of LAK to tumor targets. The protein tyrosine kinase-specific inhibitor Herb A did not block p65 phosphorylation, but blocked LAK killing of multiple tumor targets at a post-binding stage. Greater than 50% inhibition of cytotoxicity was observed after a 2.5-h pretreatment with 0.125 $\mu$g/ml Herb A. Inhibition occurred over a period in pretreatment which LAK were not dependent upon IL-2 for maintenance of killing activity, supporting the conclusion that the drug interfered with mobilization of cytotoxic function. Granule exocytosis measured by BLT-esterase release from LAK occurred after coincubation with tumors, and was inhibited by Herb A LAK cytotoxicity was dependent upon extracellular calcium, suggesting that granule exocytosis rather than Fas ligand was the principal pathway leading to target cell death. The data indicate that protein tyrosine kinases play a pivotal role in LAK cytolytic function by regulating granule exocytosis, and that tumor targets can activate an adhesion dependent Ser kinase pathway in LAK resulting in phosphorylation of L-plastin. ^

Micropropagación de Cissus tiliacea, planta del sur del estado de México

Se micropropagó Cissus tiliacea, recurso fitogenético con potencial agronómico y farmacológico, en los medios de cultivo Murashige-Skoog (MS) y Lloyd y McCown (WPM). En ambos medios se generaron resultados similares para número de brotes, nudos, hojas y raíces adventicias, sólo existió diferencia significativa (p ≤ 0,05) en la formación de callo. Para la multiplicación in vitro se utilizó WPM adicionado con 0; 0,5; 1,0; 1,5 ó 2,0 mg L-1 de benciladenina (BA) y se emplearon tres tipos de segmentos nodales (basal, medio y apical). Las concentraciones de 0 y 0,5 mg L-1 de BA resultaron en un mayor tamaño y desarrollo del explante, además permitieron la formación de 1,2 a 1,6 raíces por explante. Las concentraciones de 1,5 y 2,0 mg L-1 de BA indujeron la formación de callo. No existió diferencia significativa en las variables evaluadas por efecto del tipo de segmento nodal establecido in vitro. En el enraizamiento, en el medio MS, se evaluaron tres tipos de auxinas: ácido naftalen-1-acético (ANA), ácido indol-3-butírico (AIB) y ácido indol- 3-acético (AIA) a 0,5 mg L-1; el mayor número de raíces secundarias y diámetro de la raíz principal fue inducido por ANA, sin embargo AIB indujo una mayor elongación de la raíz principal. Los resultados del presente trabajo sugieren que el cultivo in vitro de C. tiliacea es una alternativa para su conservación y multiplicación.

Quantitative micrograph, HPLC and FTIR profiles of Melissa officinalis and Nepeta cataria (Lamiaceae) from Argentina

Melissa officinalis L., so called "Melissa" or "Toronjil", is a perennial aromatic herb, whose leaves are used in traditional medicine as a carminative, digestive and sedative, both in simple as in mixtures. Meanwhile, Nepeta cataria L., commonly called "Cat mint" or "Toronjil", with some similar properties, often replacing M. officinalis in the market, although their chemical composition is not completely matched, and contains an iridoid potentially toxic (nepetalactone). It is therefore necessary to establish diacritic parameters to differentiate these species, both at crude drug level, mixtures and extracts. Samples from various sources in Argentina were studied and documental specimens are preserved in the Herbarium UNSL. Anatomical sections were analyzed, and quantitative micrographic parameters were obtained, together with HPLC and FTIR spectra from methanolic and aqueous lyophilized extracts. Significant differences were detected in the prevailing smell foliage, morphology of leaves and inflorescences, trichomata type, palisade ratio, veinlet termination number, rosmarinic acid concentration (with distinctive HPLC profiles), and the CO/CH relationships obtained by FTIR from the extracts, that together allow adequate differentiation of both drugs, even when they were ground or powdered.

Pollen record and age determination of a sediment profile from Lake Teletskoye, Russia

A high-resolution pollen record from Lake Teletskoye documents the climate-related vegetation history of the northern Altai Mountain region during the last millennium. Siberian pine taiga with Scots pine, fir, spruce, and birch dominated the vegetation between ca. AD 1050 and 1100. The climate was similar to modern. In the beginning of the 12th century, birch and shrub alder increased. Lowered pollen concentrations and simultaneous peaks in herbs (especially Artemisia and Poaceae), ferns, and charcoal fragments point to colder and more arid climate conditions than before, with frequent fire events. Around AD 1200, regional climate became warmer and more humid than present, as revealed by an increase of Siberian pine and decreases of dry herb taxa and charcoal contents. Climatic conditions were rather stable until ca. AD 1410. An increase of Artemisia pollen may reflect slightly drier climate conditions between AD 1410 and 1560. Increases in Alnus, Betula, Artemisia, and Chenopodiaceae pollen and in charcoal particle contents may reflect further deterioration of climate conditions between AD 1560 and 1810, consistent with the Little Ice Age. After AD 1850 the vegetation gradually approached the modern one, in conjunction with ongoing climate warming.

Pollen record and age determinations on four sites from Taymyr Peninsula, Russia

Pollen data from a Levinson-Lessing Lake sediment core (74°28'N, 98°38'E) and Cape Sabler, Taymyr Lake permafrost sequences (74°33'N, 100°32'E) reveal substantial environmental changes on the northern Taymyr Peninsula during the last c. 32 000 14C years. The continuous records confirm that a scarce steppe-like vegetation with Poaceae, Artemisia and Cyperaceae dominated c. 32 000-10 300 14C yr BP, while tundra-like vegetation with Oxyria, Ranunculaceae and Caryophyllaceae grew in wetter areas. The coldest interval occurred c. 18 000 yr BP. Lateglacial pollen data show several warming events followed by a climate deterioration c. 10 500 14C yr BP, which may correspond with the Younger Dryas. The Late Pleistocene/Holocene transition, c. 10 300-10 000 14C yr BP, is characterized by a change from the herb-dominated vegetation to shrubby tundra with Betula sect. Nanae and Salix. Alnus fruticosa arrived locally c. 9000-8500 14C yr BP and disappeared c. 4000-3500 14C yr BP. Communities of Betula sect. Nanae, broadly distributed at c. 10 000-3500 14C yr BP, almost disappeared when vegetation became similar to the modern herb tundra after 3500-3000 14C yr BP. Quantitative climate reconstructions show Last Glacial Maximum summer temperature about 4°C below the present and Preboreal (c. 10 000 14C yr BP) temperature 2-4°C above the present. Maximum summer temperature occurred between 10 000 and 5500 14C yr BP; later summers were similar to present or slightly warmer.

Pollen record and age determination of a sediment profile from Frengstadsetra, Norway

Innerdalen was once a mountain valley (ca. 780 m a.s.l.) with birch forests, bogs and several summer farms. Today it is a 6.5 km**2 artifical lake. In 1980 and 1981 archaeological and palynological investigations were carried out due to the hydroelectric power plans. Radiocarbon dated pollen diagrams from 9 different localities in Innerdalen provide information on a mountain environment which has been exploited to varying degrees by human groups for thousands of years. In the Birch Zone, ca. 9500-8500 years B.P., the deglaciated surface is vegetated by the normal sequence of pioneering species, first show-bed communities, then shrub/dwarf-shrub communities, and finally a birch forest community. In the Pine Zone, ca. 8500-7500 years B.P., the mixed Birch-Pine forest which prevailed at the end of the Birch Zone is replaced by a dense pine forest. The tree limit was higher than it is today. In the Alder Zone, ca. 7500-4000 years B.P., the newly arrived alder gradually succeeded pine, particularily on good soils. This alder forest has a modem analog in the pre-alpine gray alder forests in Norway. In the last part of the Alder Zone, ca. 6000-4000 years B.P., elm and hazel are nominally present on particularily rich soils, marking the edaphic and climatic optimum in Innerdalen. During this time the first evidence of human impact on the vegetation is apparent in the pollen diagrams. At both Sætersetra in the south of the valley and Liabekken in the north, forest clearance and the development of grazed grass meadows is documented, and human impact continues until the present. The Herb Zone, ca. 4000 years B.P. to 1600 A.D., is characterized by the rapid decline of alder. The forest is increasingly open, and bog formation is initiated. The sub-alpine belt of birch forest is established, probably due to the shift to a cooler, moister climate. Human activity can also have influenced the vegetational changes, although at 4 of the localities human activity also is first apparent after the alder decline. Some localities show measurably less human impact on the vegetation ca. 2600-2000 years B.P. Grazing intensity increases ca. 2000 years B.P. At the end of the Herb Zone rye and barley pollen is registered at Sætersetra and Flonan, indicating contact between the grazing activities of Innerdal and grain cultivation activities outside the valley. The Spruce Zone, ca. 1600 A.D. to the present, does not begin synchronously since the presence of long-distance transported spruce pollen at a locality is entirely dependent on the density of the vegetation ie. degree of human impact. The youngest spruce rise is ca. 1500 A.D. at Røstvangen, when summerfarming is initiated. Summerfarming activities in Innerdal produce an increasingly open landscape. Rye and barley pollen at several localities may indicate limited local cultivation, but is more likely long-distance transport via humans and domesticated animals from cultivated areas outside Innerdalen.