Ultrasound-Assisted "Green" Extraction (UAE) of Antioxidant Compounds (Betalains and Phenolics) from Opuntia stricta var. Dilenii's Fruits: Optimization and Biological Activities.

Opuntia stricta var. Dillenii's prickly pears are an underutilized fruit with a high content of betalains and phenolic compounds that could bring potential health benefits for humans. The aim of this study is the optimization of the "green" extraction of betalains and phenolic compounds from Opuntia stricta var. Dillenii's whole fruits by ultrasound-assisted extraction (UAE), using a response surface methodology (RSM) by a central composite design (CCD) in order to obtain extracts rich in betalains and phenolic compounds with proven biological activities. For UAE optimization, the extraction temperature (20-50 °C), the amplitude (20-50%) and the ethanol volume in extraction solvent (15-80%, v/v) were selected as independent variables. All combinations were conducted at 2, 5, 10, 20 and 30 min to determinate the time effect. The betalain and phenolic compound content in Opuntia stricta var. Dillenii's whole fruits and UAE extracts were identified by HPLC-DAD-ESI/MS and HPLC-DAD-MS/QTOF and the antioxidant (ORAC method) and the anti-inflammatory (hyaluronidase inhibition method) in vitro biological activities also were determined. The most efficient extraction time was 5 min and the best UAE parameter combination was 50% amplitude, 15% ethanol in solvent (ethanol/water, 15/85, v/v) and 20 °C temperature, obtaining 10.06 ± 0.10 mg of total major betalains/g dry weight, 2.32 ± 0.08 mg of piscidic acid/g dry weight and 0.38 ± 0.00 mg of total major flavonoids/g dry weight. All applied UAE combinations significantly improved the in vitro bioactive activities (antioxidant and anti-inflammatory) of the Opuntia stricta var. Dillenii's extracts compared to the bioactivities of the extracts obtained by standard homogenization processes.

Characterization, Stability, and Bioaccessibility of Betalain and Phenolic Compounds from Opuntia stricta var. Dillenii Fruits and Products of Their Industrialization.

The aim of the present study was the full characterization, quantification, and determination of the digestive stability and bioaccessibility of individual betalain and phenolic compounds of Opuntia stricta, var. Dillenii fresh fruits (peel, pulp, and whole fruit) and of the products of the industrialization to obtain jam (raw pressed juice (product used for jam formulation), by-product (bagasse), and frozen whole fruit (starting material for jam production)). Opuntia stricta var. Dillenii fruits and products profile showed 60 betalain and phenolic compounds that were identified and quantified by HPLC-DAD-ESI/MS and HPLC-DAD-MS/QTOF, being 25 phenolic acids (including isomers and derivatives), 12 flavonoids (including glycosides), 3 ellagic acids (including glycosides and derivative), and 20 betanins (including degradation compounds). In vitro gastrointestinal digestion was performed by INFOGEST® protocol. Fruit pulp showed the greater content of total betalains (444.77 mg/100 g f.w.), and jam only showed very low amounts of two betanin degradation compounds, Cyclo-dopa-5-O-ß-glucoside (and its isomer) (0.63 mg/100 f.w.), and two Phyllocactin derivatives (1.04 mg/100 g f.w.). Meanwhile, fruit peel was the richer tissue in total phenolic acids (273.42 mg/100 g f.w.), mainly in piscidic acid content and total flavonoids (7.39 mg/100 g f.w.), isorhamnetin glucoxyl-rhamnosyl-pentoside (IG2) being the most abundant of these compounds. The stability of betalains and phenolic compounds during in vitro gastrointestinal digestion is reported in the present study. In Opuntia stricta var. Dillenii pulp (the edible fraction of the fresh fruit), the betanin bioaccessibility was only 22.9%, and the flavonoid bioaccessibility ranged from 53.7% to 30.6%, depending on the compound. In non-edible samples, such as peel sample (PE), the betanin bioaccessibility was 42.5% and the greater bioaccessibility in flavonoids was observed for quercetin glycoside (QG1) 53.7%, the fruit peel being the most interesting material to obtain antioxidant extracts, attending to its composition on antioxidant compounds and their bioaccessibilities.

Carotenoid and Carotenoid Ester Profile and Their Deposition in Plastids in Fruits of New Papaya (Carica papaya L.) Varieties from the Canary Islands.

The carotenoid profile of non-saponified and saponified extracts of different tissues (pulp and peel) of fruits of three new papaya varieties, Sweet Mary, Alicia, and Eksotika, was characterized for the first time, and almost all carotenoid compounds were quantified. Carotenoids and carotenoid esters were analyzed and characterized using HPLC-photo diode array (PDA-MS with atmospheric pressure chemical ionization with positive ion mode (APCI+) with a C30 reversed-phase column. The carotenoid deposition in collenchyma and chlorenchyma cells of papaya pulp and peel tissues was assessed by optical microscopy, confocal laser scanning microscopy, and transmission electron microscopy. The most abundant carotenoids in the fruit of the three papaya varieties (pulp and peel) were (all-E)-lycopene (230.0-421.2 µg/100 g fresh weight), (all-E)-ß-carotene (120.3-233.2 µg/100 g fresh weight), and (all-E)-ß-cryptoxanthin laurate (74.4-223.2 µg/100 g fresh weight. Moreover, high concentrations of (all-E)-lutein (922.5-1381.1 µg/100 g fresh weight) and its esters, such as (all-E)-lutein-3-O-myristate and (all-E)-lutein dimyristate, were found in peel extracts. The optical microscopy study of papaya pulps showed that carotenoid deposition in all papaya varieties, including Maradol, was mainly localized close to the cell walls, showing the presence of some crystalloids and round-shaped structures, with different sizes and distribution due to the different carotenoid content among varieties. No crystalloids or globular depositions were found in any of the peel sections, and no remarkable differences were found in the papaya peel microstructure of the different papaya varieties.

Anti-cancer stem cell activity of a sesquiterpene lactone isolated from Ambrosia arborescens and of a synthetic derivative.

New regimens are constantly being pursued in cancer treatment, especially in the context of treatment-resistant cancer stem cells (CSCs) that are assumed to be involved in cancer recurrence. Here, we investigated the anti-cancer activity of sesquiterpene lactones (SLs) isolated from Ambrosia arborescens and of synthetic derivatives in breast cancer cell lines, with a specific focus on activity against CSCs. The breast cancer cell lines MCF-7, JIMT-1, and HCC1937 and the normal-like breast epithelial cell line MCF-10A were treated with the SLs damsin and coronopilin, isolated from A. arborescens, and with ambrosin and dindol-01, synthesized using damsin. Inhibitory concentration 50 (IC50) values were obtained from dose-response curves. Based on IC50 values, doses in the µM range were used for investigating effects on cell proliferation, cell cycle phase distribution, cell death, micronuclei formation, and cell migration. Western blot analysis was used to investigate proteins involved in cell cycle regulation as well as in the NF-κB pathway since SLs have been shown to inhibit this transcription factor. Specific CSC effects were investigated using three CSC assays. All compounds inhibited cell proliferation; however, damsin and ambrosin were toxic at single-digit micromolar ranges, while higher concentrations were required for coronopilin and dindol-01. Of the four cell lines, the compounds had the least effect on the normal-like MCF-10A cells. The inhibition of cell proliferation can partly be explained by downregulation of cyclin-dependent kinase 2. All compounds inhibited tumour necrosis factor-α-induced translocation of NF-κB from the cytoplasm to the nucleus. Damsin and ambrosin treatment increased the number of micronuclei; moreover, another sign of DNA damage was the increased level of p53. Treatment with damsin and ambrosin decreased the CSC subpopulation and inhibited cell migration. Our results suggest that these compounds should be further investigated to find efficient CSC-inhibiting compounds.

Inhibition of Protein Kinases AKT and ERK1/2 Reduce the Carotid Body Chemoreceptor Response to Hypoxia in Adult Rats.

The carotid body is the main mammalian oxygen-sensing organ regulating ventilation. Despite the carotid body is subjected of extensive anatomical and functional studies, little is yet known about the molecular pathways signaling the neurotransmission and neuromodulation of the chemoreflex activity. As kinases are molecules widely involved in motioning a broad number of neural processes, here we hypothesized that pathways of protein kinase B (AKT) and extracellular signal-regulated kinases ½ (ERK1/2) are implicated in the carotid body response to hypoxia. This hypothesis was tested using the in-vitro carotid body/carotid sinus nerve preparation ("en bloc") from Sprague Dawley adult rats. Preparations were incubated for 60 min in tyrode perfusion solution (control) or containing 1 µM of LY294002 (AKT inhibitor), or 1 µM of UO-126 (ERK1/2 inhibitor). The carotid sinus nerve chemoreceptor discharge rate was recorded under baseline (perfusion solution bubbled with 5 % CO(2) balanced in O(2)) and hypoxic (perfusion solution bubbled with 5 % CO(2) balanced in N(2)) conditions. Compared to control, both inhibitors significantly decreased the normoxic and hypoxic carotid body chemoreceptor activity. LY294002- reduced carotid sinus nerve discharge rate in hypoxia by about 20 %, while UO-126 reduces the hypoxic response by 45 %. We concluded that both AKT and ERK1/2 pathways are crucial for the carotid body intracellular signaling process in response to hypoxia.

Multiple anticancer effects of damsin and coronopilin isolated from Ambrosia arborescens on cell cultures.

Terpenoids in plants are important sources for drug discovery. In this study, we extracted damsin and coronopilin, two sesquiterpene lactones, from Ambrosia arborescens and examined their anticancer effects on cell cultures. Damsin and coronopilin inhibited cell proliferation, DNA biosynthesis and formation of cytoplasmic DNA histone complexes in Caco-2 cells, with damsin being more potent than coronopilin. Further studies using the luciferase reporter system showed that damsin and coronopilin also inhibited expressions of nuclear factor-κB (NF-κB) and signal transducer and activator of transcription-3 (STAT3), indicating that these sesquiterpenes can interfere with NF-κB and STAT3 pathways. Finally, we examined the effects of two synthetic dibrominated derivatives of damsin, 11α,13-dibromodamsin and 11ß,13-dibromodamsin. While bromination appeared to weaken the antiproliferative effects of damsin, the ß epimer had strong inhibitory effects on STAT3 activation. In conclusion, the sesquiterpene lactones damsin and coronopilin have inhibitory effects on cell proliferation, DNA biosynthesis and NF-κB and STAT3 pathways, thus being potentially important for discovery of drugs against cancer.

Antioxidant and antimutagenic polyisoprenylated benzophenones and xanthones from Rheedia acuminata.

Dichloromethane extract of the stem bark of Rheedia acuminata yielded three benzophenones with antioxidant activity, the new one named acuminophenone A (1), guttiferone K (2) and isoxanthochymol (3), along with the known xanthones formoxanthone C (4) and macluraxanthone (5). The structures were established through interpretation of their spectroscopic data, the stereochemistry of compounds (1) and (2) were resolved by experimental and computational experiments and their antioxidant activities were measured using the DPPH, ABTS and TEAC assays. The antioxidant results showed that metabolites 1, 4 and 5 had a better antioxidant activity than the reference compound quercetin. In addition, we evaluate the mutagenicity and antimutagenicity of the CH2Cl2 extract as well as of the free radical scavenger compounds 1, 4 and 5 by the AMES Salmonella/microsomal test. No mutagenicity was found in the CH2Cl2 extract using Salmonella typhimurium strains TA98, TA100, TA102, TA1537 and TA1538, with or without S9 metabolic activation. The pure compounds neither showed mutagenicity in TA 102 strain and the most important result was the strong reduction of mutagenic effect induced by hydrogen peroxide in S. typhimurium TA102, with or without S9, showed by the compounds 1 (more than 93%) and 4 (more than 88%) at 0.02 microg/plate.

Antiproliferative effects of curcuphenol, a sesquiterpene phenol.

Curcuphenol is a sesquiterpene isolated from sponges and plants having several significant biological activities. The present study explored its effect on cell proliferation and apoptosis in Caco-2 human colon cancer cells. It was demonstrated that curcuphenol in concentrations in the range of 29-116 µg/ml inhibited cell proliferation and DNA replication and induced cell death in a dose-dependent manner. The induction of apoptosis was associated with a stimulation of the activity of caspase-3. The findings presented here suggest that curcuphenol has antiproliferative and pro-apoptotic properties.

Actividad Genotóxica de Opuntia soehrensii, evaluada por el Test de Mutación y Recombinación Somática en D. melanogaster / Genotoxic Activity of Opuntia soehrensii, evaluated by the Test of Mutation and Somatic Recombination in D. melanogaster

El extracto acuoso de Opuntia soehrensii, planta medicinal utilizada como antiviral en la farmacopea boliviana fue evaluado en su capacidad genotóxica por el test SMART en alas de D. melanogaster. Por tratarse de un extracto entero se evaluó por dos cruces diferentes; el cruce estandar y el cruce de Alta Bioactivación. Los resultados muestran que Opuntia soeherensii no es genotóxica en concentraciones de 3.57 mg/ml y 7.4 mg/ml para ninguno de los dos cruces, pero es promutagénica a 1.78 mg/ml.

Evaluación genotoxicológica del extracto etéreo de baccharis incarum / Genotoxicológica evaluation of the ethereal extract of Baccharis incarum

La genotoxidad del extracto etéreo de Baccharis incarum, una planta utilizada en la medicina tradicional boliviana, ha sido evaluada por el test de Mutación y Recombinación Somática en alas de Drosophila melanogaster. Este extracto no es genotóxico de acción indirecta.

Genotoxicidad/ antigenotoxicidad de Bacharis latifolia / Genotoxicity/antigenotoxicity of Bacharis latifolia

El potencial genotóxico y antigenotóxico del extracto diclorometánico de Bacharis latifolia fue evaluado por el Test de Mutación y Recombinación Somática (SMART) en alas de Drosophila melanogaster usando dos diferentes cruces. Los resultados muestran que B.latifolia no es genotóxica ni antigenotóxica. La presencia de flavonoides en su composición química corroboran su actividad fotoprotectora.

Evaluación Genotoxicológica de suelos expuestos a plaguicidas por el test de micronúcleos / Evaluation genotoxicológica de suelos expuestos to pesticides by the test of micronucleuses

En el presente estudio se evaluó la composición físicaquímica y la capacidad genotoxica de suelos contaminados de la comunidad de Huaraco, Dpto. deLa Paz por el test de Micronúcleos en Vicia faba. Los resultados muestran que el suelo de Huaraco expuesto hace 20 años a pesticidas no es genotóxico, mientras que el suelo expuesto recientemente muestra un incremento en su capacidad genotóxica.

Epidemiology and risk factors of eating disorders: a two-stage epidemiologic study in a Spanish population aged 12-18 years.

OBJECTIVE: The authors studied the prevalence rates of eating disorders (ED) and their risk factors in a Spanish population aged 12-18 years. METHOD: A two-stage epidemiologic study was conducted in the province of Valencia. Educational centers, classrooms, and individuals were selected randomly. The initial sample comprised 544 subjects. During Stage 1, subjects were screened with the 40-item Eating Attitude Test and a sociodemographic questionnaire that evaluates risk factors. During Stage 2, a semistandardized clinical interview was conducted with each participant. A random control group was paired by class, age, and sex. Comorbid psychiatric disorders and partial and subclinical forms were detected using criteria in the 4th ed. of the Diagnostic and Statistical Manual of Mental Disorders (DSM-IV). RESULTS: Using DSM-IV criteria, the morbidity rate was 2.91%, women comprised 5.17% of the sample, men comprised 0.77% of the sample, and the sex ratio was 6.71. Using DSM-IV criteria, including subclinical forms, the morbidity rate was 5.56%, women comprised 10.3% of the sample, men comprised 1.07% of the sample, and the sex ratio was 9.63. Six risk factors were isolated: psychiatric comorbidity, friend on a diet in the last year, desire to lose weight, desire to be less corpulent, sentimental problems, and diet in the last year. DISCUSSION: This is the first Spanish two-stage epidemiologic study to include a control group and to investigate risk factors.

Comunicación de la segunda hemoglobina mutada en Bolivia "Hb Illimani" / Report of the second mutated hemoglobin in Bolivia \"Illimani Hb\"

En 112 pacientes eritrocitémicos, se encontró la segunda variante de Hb en Bolivia, correspondiente a una beta-mutada, de migración en agar y menos estable que la Hb5, por sus características y ubicación, es una probable nueva Hb mutada

Comunicacion de la segunda hemoglobina mutada en Bolivia "Hb Illimani" / Communication of the secon hemoglobin mutation in Bolivia

Por screening de hemoglobinas, en una señora de 66 años que presenta eritrocitosis, se encuentra la segunda variante de Hb en Bolivia, corresponde a una beta mutada, de migracion lenta, de migracion en agar y menos estable que la HbF y HbS. Por sus caracteristicas y ubicacion, es una probable nueva Hb