Cardiovascular diseases account for the majority of deaths worldwide. Many of their risk factors have been identified but, for their continued study, research centering on new murine models is of interest. In this study, a high fat diet (HFD) and a normal diet (ND) (25 and 4.4% fat, respectively) were tested over a 40-day period to induce the same metabolic alterations in CF-1 mice in two separate experiments. The parameters measured for these effects corresponded to the weight of ingested food and water, to the weight of the mice and their selected organs (adipose tissue, gastrocnemius, liver and heart), to their biochemical profile (glycemia, blood uric nitrogen, uric acid, triglycerides, cholesterol, proteins and albumin) and to the percentage of fat in their livers. The biochemical profile of the CF-1 mice fed a diet high in fat but balanced in proteins (16.9%) showed statistically significant increases in glycemia, cholesterol and triglyceride levels. A statistically significant increase in the weight of adipose tissue was also observed. No statistically significant differences were observed in the muscular mass of either of the groups of mice, but a high percentage of fat was found in the liver. The results lead to the conclusion that CF-1 mice fed a HFD develop metabolic alterations that correspond to an equivalent metabolic syndrome. This is important in the evaluation of the effects of various interventions, such as food, exercise and molecules, on metabolic alterations in mice induced by the intake of a HFD.

An infusion or decoction of the aerial parts of Fabiana imbricata (Solanaceae) is used in traditional medicine in Chile and Argentina as a digestive and diuretic agent. The main sesquiterpene of the plant was identified as 11-hydroxy-4-amorphen-15-oic acid. At doses of 25, 50 and 100 mg/kg, the compound showed a dose-dependent gastroprotective effect in HCl/EtOH-induced gastric lesions in mice reducing the lesions by 68% at 100 mg/kg. Seven derivatives of the terpene were prepared and their gastroprotective effect was assessed in HCl/EtOH-induced gastric lesions in mice. The cytotoxicity of the products was evaluated in fibroblasts and AGS cells. At 100 mg/kg, 11-hydroxy-4-amorphen-15-p-toluidinamide presented the best gastroprotective effect reducing the gastric lesions by 80%, showing a similar effect to lansoprazole at 20 mg/kg. The compound, however, presented higher cytotoxicity than other derivatives with the IC50 ranging between 1.10 and 145 mu M in AGS cells and fibroblasts, respectively. Most compounds proved to be non-toxic showing cytotoxicity values higher than 1000 mu M. The spectroscopic data of six 11-hydroxy-4-amorphen-15-oic acid derivatives are presented here for the first time. Copyright (c) 2005 John Wiley & Sons, Ltd.

Boldo (Peumus boldus) is a medicinal tree or shrub native to the Mediterranean climatic zone of Chile. The perennial leaves are used in medicine for their digestive, choleretic, and liver-protection properties. The total production of 1,500 t dried leaves per year is wild crafted. Three different boldo populations were chosen for selection purposes and their progenies were studied for alkaloid and essential oil production. Thirty individuals were analyzed in each wild population in 1996 and 2002. No differences in essential oil concentration (1.3-1.6 ml/100 g dry weight (DW) were found for these populations. Progenies were grown for five years under the same field conditions and analyzed over the last three years. The progeny of the northern population had significantly higher essential oil content than the others (3.5 ml/100 g DW and 2.0-2.6 ml/100 g DW, respectively) and showed the highest alkaloid concentration in its natural habitat (0.34% versus 0.20 and 0.22%, respectively) but progenies did not differ in alkaloid content (0.17-0.20%). Wild populations had significantly higher alkaloid concentrations than the cultivated progenies but lower essential oil content. The progenies of the northern and central population showed differences in essential oil content between half-sib families. As boldo is a dioecious species with cloning difficulties, populations rather than individuals should be selected. Results of this study suggest that future cultivation of boldo use seeds from populations with the highest essential oil and alkaloid concentration.

The labdane diterpene solidagenone 1 and its semisynthetic and biotransformation derivatives 2–10 were assessed for gastroprotective and ulcer-healing effect using human epithelial gastric cells (AGS) and human lung fibroblasts (MRC-5). The ability of the compounds to protect the AGS cells against the damage induced by sodium taurocholate (NaT), to stimulate the cellular reduced glutathione (GSH), prostaglandin E2 content, enhance AGS and MRC-5 cell proliferation and to scavenge superoxide anion in vitro was studied. The cytotoxicity of the compounds was assessed towards MRC-5 fibroblasts and AGS cells. A significant reduction of cell damage after NaT incubation was observed when the AGS cells were pretreated with compounds 2 and 6. Treatment with compounds 4–6, 8 and 9 significantly stimulated the GSH content in AGS cell cultures. None of the studied compounds was active as a superoxide anion scavenger. In AGS cells treated with compounds 1–10, only compound 5 was able to increase prostaglandin content. Concerning the proliferation assays, a significant stimulating effect was observed for compounds 2, 8, 9 on AGS cells and for 5, 7–9 on MRC-5 fibroblasts. Regarding cytotoxicity, solidagenone showed higher toxicity while compounds 4 and 7 were the less toxic. Our results showed that most of the studied compounds act in vitro as gastroprotectors increasing the cellular GSH content. Additionally, some derivatives exhibited in vitro ulcer-healing effect stimulating the cell proliferation.

Canelo (Drimys winteri) is a Chilean native tree with medicinal properties and sacred to mapuche people. This species contains chemical compounds with biological activity such as: essential oils, terpenes and flavonoids, which were quantified in leaves of five D. winteri and one D. andina populations by hydrodistillation (essential oils), hexane Soxhlet extraction (terpenes), and spectrophotometry (flavonoids). Mean concentrations of 0.5 mL 100 g(-1) for essential oils, 7.1 % for terpenes and 1.8 % for flavonoids in leaves (dry mass) were determined. Despite the great variation found, significant differences among populations were detected in concentrations of all groups of chemical compounds studied. Populations from the central region of Chile presented the highest values of essential oils (0.68 mL 100 g(-1) in Huerta de Maule population). terpenes, and flavonoids (9.47 and 2.37 %, respectively, both in the Pangal population). The Southernmost population, located in Chiloe Island, showed the lowest values for the three groups of compounds: 0.22 mL 100 g(-1) for essential oils, 3.36 % for terpenes and 1.21 % for flavonoids. D. winteri and D. andina populations from the IX region, geographically close to each other, had different concentrations of essential oils and flavonoids. It is concluded that different Drimys populations actually contain different quantities of essential oils, terpenes, and flavonoids.

Buddleja globose commonly known as “matico” is a medicinal shrub native to Chile. Its leaves are traditionally used for healing wounds and ulcers. The plant is collected from the wild and then marketed. To avoid over-exploitation of the natural populations, domestication studies have been started. The plant proved to be an easy-rooting species. The best percentages (up to 83 %) were observed in spring with hormonal treatments of 500 to 2,000 ppm IBA. The highest flavonoid concentrations were found in adult leaves (0.51 %), followed by young leaves (0.37 %). Concentrations of flavonoids, tannins, and alkaloids showed maximum values in spring (0.38, 1.07, and 0.012 %, respectively. Although the species is adapted to dry conditions, irrigated plants showed higher leaf yield, leaf area and tannin concentration than plants grown under water stress. INTRODUCTION Buddleja globose commonly known as “matico” is a medicinal shrub native to Chile. Traditionally, leaves are used for healing wounds and ulcers (Montes and Wilkomirsky, 1987, Mellado et al., 1996). Montes and Wilkomirsky (1987) reported tannins, alkaloids, and flavonoids as some of the active compounds. Matico leaves are collected from the wild and then marketed; however, this process involves overexploitation and a decrease in genetic diversity of the wild population. To overcome these difficulties, domestication studies of this species have been initiated, including studies on propagation, variability of alkaloids, flavonoids, and tannins, and cultivation in order to determine the effects of different irrigation and harvest treatments. MATERIAL AND METHODS Propagation For vegetative propagation, different treatments of the root cuttings were studied, including factors such as season (spring or autumn), application of auxins at different concentrations (from 0 to 3,000 ppm IBA), rooting medium (sand, perlite, and vermiculite), origin of the cuttings (tip, middle, and basal shoot portions), and wounding. The first two factors were studied using a completely randomized design with 4 replications of 50 cuttings each, whereas the studies of the other factors included 20 cuttings per replication. Studies were done in 1997 in Talca, under greenhouse conditions, irrigated by an intermittent mist system, with rooting media temperatures of 25° C in the day and 15° C during the night. Active Agents Variability of flavonoid, alkaloid and tannin concentrations was studied as Proc. Int. Conf. on MAP Eds. J. Bernáth et al. Acta Hort. 576, ISHS 2002 204 function of leaf age (senescent, adult, or young leaves) and seasonal effects (collection in spring, summer, or autumn). Samples were taken in three different locations (plants grown in nursery, plantings, and wild plants) analyzing three, eight, and five individuals, respectively, at 4 and 5 different harvest times. Leaves were dried at 40-45o C and ground. Flavonoids, alkaloids, and tannins, were quantified photometrically with modified methods described by Franz and Koehler (1992). Irrigation For the irrigation studies different origins of matico, including individuals from three wild populations and three cultivated clones, were placed in a split-plot design, one fully irrigated, the other with water stress (65 and 20 % field capacity, respectively). The origins were distributed randomly within these plots. In 50 % of the plants in each plot, the main shoot was cut. RESULTS AND DISCUSSION Rooting of the Cuttings Matico is an easy-rooting species. Best percentages (up to 83 %) were observed in spring with hormonal treatments of 500 to 2,000 ppm IBA (Table 1). Survival rates seemed to be better in autumn than in springtime, whereas in rooting, no significant differences could be observed, with the exception of the treatment with 1,000 ppm. In comparing the position of the cuttings, wounding, and rooting medium, no differences in rooting between treatments could be found after 11 weeks (Table 2), but cuttings taken from the tip of the shoots rooted faster than those taken from the middle part. At the same time, cuttings grown in sand and in a mixture of perlite and vermiculite showed better survival rates and faster root formation than those in perlite alone. Variability of Active Compounds Highest flavonoid concentrations were found in adult leaves (0.51 %), followed by young leaves (0.37 %). No significant differences among leaf ages could be observed in tannin and alkaloid concentrations (Table 3). Concentrations of flavonoids, tannins, and alkaloids showed maximum values in spring (0.38, 1.07, and 0.012 %, respectively) followed by autumn. Considering that matico leaves are growing in spring it is not recommended to harvest them at that time, even though they showed highest concentrations of active compounds, they had low yields of dry matter. In autumn, concentrations of flavonoids and alkaloids are not significantly different from those in spring, but there is an increase in leaf-yield, and adult leaves are richer in flavonoid concentration than young ones. Late harvests should be avoided to prevent high percentages of senescent leaves in the product. Effect of Water Stress Although the species is adapted to dry conditions, irrigated plants showed higher leaf yield, leaf area and tannin concentration than plants grown under water stress (Table 4). Regarding the origin of plant material, no significant differences could be observed in leaf yield, leaf area, flavonoid and tannin concentration, only in alkaloids and the proportion of leaf: total yield. This indicates that cultivated plants have not previously been selected for the studied characters. ACKNOWLEDGEMENTS We wish to thank FIA (Fundación para la Innovación Agraria), Chile, for financial support (Project Numbers: C-96-1-S-007 and V99-0-S-032), as well as Paula Jeldres, Benita González, and Andrea Pincheira for their collaboration in these studies.

The labdane diterpene solidagenone and four semisynthetic derivatives were assessed for effects on the glucocorticoid-mediated signal transduction. Solidagenone and the derivatives proved to be active with IC50 values between 1 and 25 micrograms/mL. All compounds were cytotoxic towards L 1210, BHK and COS 7 cells with IC50 from 10-100 micrograms/mL.

Boldo is a tree or shrub with medicinal properties native to Chile. The leaves contain alkaloids and essential oils. Variation of total alkaloid concentration, of the alkaloid boldine, and essential oil components were studied in different populations from northern, central, and southern parts of its geographic range and in their progenies (half-sib families). Total alkaloid concentration showed genetic variation between progenies of the central population but not between populations. Boldine content found in concentrations of 0.007 to 0.009% did not differ significantly between populations. Principal components of the essential oil were determined genetically, with highest values for ascaridole in the population of the north and for P-cymene in the south. Between half-sib families genetic variation was found in the central and northern populations for these components. The high heritability coefficients found indicate considerable potential for successful selection of individuals for these characters.

Lemon verbena is an aromatic shrub native to Chile, Argentina, and Peru. Its leaves have a pleasant odour, similar to that of lemon, due to the content of essential oils.The effects of the time of year and the time of day of harvesting on essential oil concentrations, yield, percentage of the principle component citral, and leaf area were studied in a two-year-old plantation with 6,667 plants /ha in Talca, central region of Chile (35 degrees S). Samples were taken every two weeks from October (springtime) to April (autumn). To study the fluctuations of essential oil concentration during the day, samples were taken every three hours from 7.30 a.m. to 7.30 p.m. on five different days between December and April.Maximum essential oil concentration was found in young leaves in October with 0.95 ml/100 g DW, decreasing to values of 0.09 to 0.20 ml/100 g DW from December to April. Essential oil yield increased from October to March, reaching a maximum of 27.3 l/ha. Citral content was highest in November (up to 64%), than decreased to reach a minimum in December (34%). From February Values maintained stable (52-45%). In November largest leaf areas were found with 9.2 cm(2)/leaf.No differences in essential oil concentration between different times of day of harvesting could be found. Citral content increased during the day.

The activity of jatrogrossidione, the main diterpene of Jatropha grossidentata and jatrophone from Jatropha isabellii was determined against Leishmania and Trypanosoma cruzi strains in vitro as well as against Leishmania amazonensis in vivo. Jatrogrossidione showed a strong in vitro leishmanicidal and trypanocidal activity with IC100 of 0.75 and 1.5–5.0 μg/mL, respectively. Under similar conditions, the IC100 of glucantime, ketoconazole and pentamidine towards Leishmania strains were >100, 50–100 and 1 μg/mL, respectively. The IC50 of jatrogrossidione was <0.25 μg/mL against amastigote forms of Leishmania infecting macrophages, with toxicity at concentrations higher than 0.5 μg/mL. BALB/c mice infected with L. amazonensis strain PH 8 were treated 24 h after infection with jatrogrossidione and jatrophone for 13 consecutive days. Jatrophone at 25 mg/kg/day subcutaneously administered was significantly active (p<0.05) against the virulent strain PH 8 of L. amazonesis; it was more active than Glucantime at 112 mg Sbv per kg/day. Subcutaneous administration of jatrophone, however, proved to be too toxic under our assay conditions. Assays of single local treatment on the footpad infection 2 weeks after inoculation of L. amazonensis indicated that jatrogrossidione and jatrophone were inactive at the selected doses.

Equisetum bogotense (Fquisetaceae) and Fuchsia magellanica (Onagraceae) were used by the Mapuche Amerindians as diuretics. Intravenous administration of hydroalcoholic extracts to rats elicited a hypotensive response of -29.0%+/-4.5% and -24.2%+/-0.5% respectively in the mean blood pressure of normotensive animals at a dose of 5 mg crude extract/kg body weight. The activity was found to be related with polar compounds. For Fuchsia, the active principles are related to tannins. A single oral dose of 500 mg/kg body weight Equisetum extract produced a significant increase (p < 0.05) in the urine output in rats, while in Fuchsia a reduction in diuresis was observed. The diuretic effect of the Equisetum extract was weak compared with hydrochlorothiazide at 25 mg/kg body weight. The data presented support the use of E. bogotense in Chilean traditional medicine. A micropropagation method for E. bogotense was established by culturing shoot tips in Murashige-Skoog medium with macro and micronutrients diluted 4-fold and containing 0.1 mg/L thiamine, 100 mg/L myoinositol, 1.0 mg/L NAA and 0.1 mg/L BAP.

An HPLC method for the simultaneous quantitative determination of rutin and scopoletin in the aerial parts of Fabiana imbricata is presented. The results showed high variability in the scopoletin (240-2,400 mg%) and rutin (195-1,950 mg%) content in the populations surveyed. A micropropagation method for F. imbricata plants was established by culturing shoot tips on Murashige-Skoog (MS) medium containing 1.0 mg/l benzylaminopurine (BAP), 0.01 mg/l naphthaleneacetic acid (NAA), and 0.1 mg/l gibberellic acid (GA3). Auxin addition to the rooting medium, especially 0.5 mg/l indolebutyric acid (IBA) or 0.5 mg/l IBA and 0.1 mg/l NAA, enhances root formation. The micropropagation method presented allowed the obtention of regenerated plantlets in six weeks from shoot tips.