Study on the mechanisms of Suaeda rigida polysaccharides on the heart inhibition and skeletal muscle promotion in the frog / [Estudo sobre os mecanismos dos polissacarídeos de Suaeda rigida na inibição do coração e promoção da musculatura esquelética no sapo]

The objectives of this study were to investigate the effects of different concentrations of Suaeda rigida polysaccharides (SRPs) on the physiological characteristics of the frog heart and gastrocnemius muscle, compare their similarities and differences, and analyze the mechanisms. CaCl2 and acetylcholine (Ach) were selected respectively to be co-incubated with the high concentration SRPs to observe the effects on the heart contraction of frog. The effects of different concentrations of the SRPs on the activities of acetylcholinesterase (A-CHE), Na+-K+-ATPase and Ca2+-Mg2+-ATPase in the isolated frog heart were detected by UV-Vis spectrophotometry. The gastrocnemius muscle was immersed in the high concentration of SRPs for 10 min, and the systolic indexes were recorded. The effects of SRPs on the Ach content and A-CHE activity at the sciatic nerve-gastrocnemius junction were determined by UV-Vis spectrophotometry and enzyme-linked immunosorbent assay (ELISA). The results showed that the SRPs had significant inhibitory effects on the contractile amplitude of isolated heart and the contractile amplitude induced by CaCl2 and Ach, respectively (P<0.01). The activity of Ca2+-Mg2+-ATPase was significantly promoted, and the activity of A-CHE was significantly inhibited (P<0.01). The contraction amplitude, contraction rate, relaxation rate of gastrocnemius muscle and the Ach content at the junction of sciatic nerve-gastrocnemius muscle were significantly increased (P<0.01), and the activity of A-CHE at the junction was significantly inhibited (P<0.01) by the SRPs. All the results suggested that the SRPs could inhibit the contraction of heart and promote the contraction and relaxation of skeletal muscle. The mechanism was related to blocking the fast INa channel, inhibiting the ICa-L and activating the M receptors of myocardial membrane and then inhibiting external Ca2+ influx, increasing Ca2+-Mg2+-ATPase activity, decreasing a-che activity.

Os objetivos deste estudo foram investigar os efeitos de diferentes concentrações de Suaeda rigida polissacarídeos (SRPs) sobre as características fisiológicas do coração de rã e do músculo gastrocnêmio, comparar suas semelhanças e diferenças, e analisar os mecanismos. CaCl2 e acetilcolina (Ach) foram selecionados respectivamente para serem co-incubados com os SRPs de alta concentração para observar os efeitos sobre a contração do coração de rã. Os efeitos das diferentes concentrações dos SRPs sobre as atividades da acetilcolinesterase (A-CHE), Na+-K+-ATPase e Ca2+-Mg2+-ATPase no coração isolado de rã foram detectados pela espectrofotometria UV-Vis. O músculo gastrocnêmio foi imerso na alta concentração de SRP por 10 min, e os índices sistólicos foram registrados. Os efeitos das SRPs no conteúdo de Ach e na atividade de A-CHE na junção nervo-gastrocnêmio ciático foram determinados pela espectrofotometria UV-Vis e pelo ensaio de imunoabsorção enzimática (ELISA). Os resultados mostraram que os SRPs tiveram efeitos inibidores significativos sobre a amplitude contrátil do coração isolado e a amplitude contrátil induzida por CaCl2 e Ach, respectivamente (P<0,01). A atividade do Ca2+-Mg2+-ATPase foi significativamente promovida e a atividade do A-CHE foi significativamente inibida (P<0,01). A amplitude de contração, a taxa de contração, a taxa de relaxamento do músculo gastrocnêmio e o conteúdo de Ach na junção do músculo nervo ciático-gastrocnêmio foram significativamente aumentados (P<0,01), e a atividade do A-CHE na junção foi significativamente inibida (P<0,01) pelas SRPs. Todos os resultados sugeriram que os SRPs poderiam inibir a contração do coração e promover a contração e o relaxamento do músculo esquelético. O mecanismo estava relacionado ao bloqueio do canal INa rápido, inibindo a ICa-L e ativando os receptores M da membrana miocárdica e depois inibindo o influxo externo de Ca2+, aumentando a atividade de Ca2+-Mg2+-ATPase, diminuindo a atividade a-che.

Myorelaxant Effect of the Dysphania ambrosioides Essential Oil on Sus scrofa domesticus Coronary Artery and Its Toxicity in the Drosophila melanogaster Model.

PURPOSE: Alternative methods for the use of animals in research have gained increasing importance, due to assessments evaluating the real need for their use and the development of legislation that regulates the subject. The principle of the 3R's (replacement, reduction and refinement) has been an important reference, such that in vitro, ex vivo and cord replacement methods have achieved a prominent place in research. METHODS: Therefore, due to successful results from studies developed with these methods, the present study aimed to evaluate the myorelaxant effect of the Dysphania ambrosioides essential oil (EODa) using a Sus scrofa domesticus coronary artery model, and the toxicity of both the Dysphania ambrosioides essential oil and its major constituent, α-terpinene, against Drosophila melanogaster in toxicity and negative geotaxis assays. RESULTS: The EODa relaxed the smooth muscle of swine coronary arteries precontracted with K+ and 5-HT in assays using Sus scrofa domesticus coronary arteries. The toxicity results presented LC50 values of 1.546 mg/mL and 2.282 mg/mL for the EODa and α-terpinene, respectively, thus showing the EODa and α-terpinene presented toxicity to these dipterans, with the EODa being more toxic. CONCLUSIONS: Moreover, the results reveal the possibility of using the EODa in vascular disease studies since it promoted the relaxation of the Sus scrofa domesticus coronary smooth muscle.

Preparative mass-spectrometry profiling of minor concentrated metabolites in Salicornia gaudichaudiana Moq by high-speed countercurrent chromatography and off-line electrospray mass-spectrometry injection.

Salicornia species have just been introduced to the European market as a vegetable named 'samphire', 'green asparagus', or 'sea asparagus'. Due to its increasing attention, and associated value, minor compounds of Salicornia gaudichaudiana Moq were investigated. The use of countercurrent chromatography and mass spectrometry enabled the search for known, as well as potentially novel natural products. Their identification was achieved based on molecular weights and mass-spectrometric fragmentation data. Low detection limits enabled the visualization of all compounds with their identification in almost real time close to the preparative countercurrent chromatography experiment. A list of known natural products from Salicornia genus guided the identification process of compounds occurring in Salicornia gaudichaudiana Moq by tandem mass spectrometry fragment comparison. The natural product classes were divided into four groups: chlorogenic acid derivatives; flavonoid derivatives; pentacyclic triterpenoid saponins; and other compounds.

Chenopodium ambrosioides associated with whole body vibration exercises alters the feed intake in Wistar rats.

The consequences of treatment involving the use of a natural product and whole body vibration (WBV) exercise have been investigated. The aim of the present study was to evaluate the effects of the joint treatment with an aqueous extract of Chenopodium ambrosioides and WBV on physiological parameters in rats. Wistar rats (n=20) were divided equally into four groups: control group (CG), treated with C. ambrosioides (CHE) group, exposed to 50 Hz of mechanial vibration (VBR), and treated with C. ambrosioides and exposed to 50 Hz of mechanical vibration (VBR + CHE) daily for 6 weeks. The body mass of the animals was determined weekly, the feed intake and the stool consistency were measured daily. One day after the 6 weeks of treatment, samples of blood were collected and used for biochemical analysis. Along 6 weeks, there was an increase (P<0.001) in the feed intake in VBR group and a decrease in the CHE group in comparison with other groups. The levels of the enzyme aspartate aminotransferase (AST) in VBR + CHE group decreased (P<0.05) in comparison with other groups. No differences were found in body mass and stool consistency. WBV altered the feed intake without directly affecting the body mass. Moreover, WBV in association with C. ambrosioides caused alteration in the enzymatic activity of AST.

Chemical Composition, Antibacterial, Schistosomicidal, and Cytotoxic Activities of the Essential Oil of Dysphania ambrosioides (L.) Mosyakin & Clemants (Chenopodiaceae).

We have investigated the chemical composition and the antibacterial activity of the essential oil of Dysphania ambrosioides (L.) Mosyakin & Clemants (Chenopodiaceae) (DA-EO) against a representative panel of cariogenic bacteria. We have also assessed the in vitro schistosomicidal effects of DA-EO on Schistosoma mansoni and its cytotoxicity to GM07492-A cells in vitro. Gas chromatography (GC) and gas chromatography-mass spectrometry (GC/MS) revealed that the monoterpenes cis-piperitone oxide (35.2%), p-cymene (14.5%), isoascaridole (14.1%), and α-terpinene (11.6%) were identified by as the major constituents of DA-EO. DA-EO displayed weak activity against Streptococcus sobrinus and Enterococcus faecalis (minimum inhibitory concentration (MIC) = 1000 µg/ml). On the other hand, DA-EO at 25 and 12.5 µg/ml presented remarkable schistosomicidal action in vitro and killed 100% of adult worm pairs within 24 and 72 h, respectively. The LC50 values of DA-EO were 6.50 ± 0.38, 3.66 ± 1.06, and 3.65 ± 0.76 µg/ml at 24, 48, and 72 h, respectively. However, DA-EO at concentrations higher than 312.5 µg/ml significantly reduced the viability of GM07492-A cells (IC50  = 207.1 ± 4.4 µg/ml). The selectivity index showed that DA-EO was 31.8 times more toxic to the adult S. mansoni worms than GM07492-A cells. Taken together, these results demonstrate the promising schistosomicidal potential of the essential oil of Dysphania ambrosioides.

Evaluation of indigenous grains from the Peruvian Andean region for antidiabetes and antihypertension potential using in vitro methods.

The health-relevant functionality of 10 thermally processed Peruvian Andean grains (five cereals, three pseudocereals, and two legumes) was evaluated for potential type 2 diabetes-relevant antihyperglycemia and antihypertension activity using in vitro enzyme assays. Inhibition of enzymes relevant for managing early stages of type 2 diabetes such as hyperglycemia-relevant alpha-glucosidase and alpha-amylase and hypertension-relevant angiotensin I-converting enzyme (ACE) were assayed along with the total phenolic content, phenolic profiles, and antioxidant activity based on the 1,1-diphenyl-2-picrylhydrazyl radical assay. Purple corn (Zea mays L.) (cereal) exhibited high free radical scavenging-linked antioxidant activity (77%) and had the highest total phenolic content (8 +/- 1 mg of gallic acid equivalents/g of sample weight) and alpha-glucosidase inhibitory activity (51% at 5 mg of sample weight). The major phenolic compound in this cereal was protocatechuic acid (287 +/- 15 microg/g of sample weight). Pseudocereals such as Quinoa (Chenopodium quinoa Willd) and Kañiwa (Chenopodium pallidicaule Aellen) were rich in quercetin derivatives (1,131 +/- 56 and 943 +/- 35 microg [expressed as quercetin aglycone]/g of sample weight, respectively) and had the highest antioxidant activity (86% and 75%, respectively). Andean legumes (Lupinus mutabilis cultivars SLP-1 and H-6) inhibited significantly the hypertension-relevant ACE (52% at 5 mg of sample weight). No alpha-amylase inhibitory activity was found in any of the evaluated Andean grains. This in vitro study indicates the potential of combination of Andean whole grain cereals, pseudocereals, and legumes to develop effective dietary strategies for managing type 2 diabetes and associated hypertension and provides the rationale for animal and clinical studies.

[Chemical composition of 2 wild vegetable species compared with chard]. / Estudio de la composición química, de dos especies vegetales silvestres en comparación con acelga.

The nutritional quality of two wild vegetable species, Kochia scoparia (Ks) and Chenopodium album (Cha) was evaluated and compared with Beta vulgaris var. cicla(chard) (Bv), in order to propose their domestication as alternative protein sources. Chemical percentual analysis (AOAC), gaseous chromatography fatty acid determination, and antinutrient searching (by chemical, enzymatic and immunological method) were performed. Protein quality was determined by the following indexes: net protein utilization (NPU), true digestibility (tD) and biological value (BV). The three species exhibited similar protein concentration values, 25.8, 25.0 and 22.1 g/100 g for Ks, Cha and Bv, respectively. Kochia scoparia showed the highest value for fatty acid analysis (82%), with predominance of linolenic acid. Since the samples were subjected to boiling, the amounts of antinutrients found can be considered to be within levels not affecting health. As regards the biological quality, the obtained values for Ks, Cha and Bv, respectively, were: NPU: 68.0 +/- 0.4, 55.0 +/- 6.1 and 56.0 +/- 4.2; tD 70.0 +/- 0.7, 71.0 +/- 4.0 and 76.0 +/- 8.2; BV: 97, 77 and 74. The analysis of the biological quality indexes gave highest nitrogen profit for Ks. These results indicate that Kochia scoparia is suitable for the objective proposed.

Estudio de la composición química, de dos especies vegetales silvestres en comparación com acelga / The study of the chemical composition of 2 wild vegetable species in comparison with chard

El propósito de este estudio fue evaluar la calidad nutricional de dos especies vegetales silvestres Kochia scoparia (Ks) y Chenoposium album (Cha) y comparalas con Beta vulgaris var. cicla (acelga) (Bv), con la finalidad de proponer la domesticación de las mismas, para introducirlas como fuente proteica alternativa. Se realizó la evaluación químico porcentual (AOAC), determinación de ácidos grasos por cromatografía gaseosa, la investigacón de antinutrientes (Métodos químicos, enzimáticos e inmunológicos) y mediante 'los índices de utilización proteica (NPU), digestibilidad verdadera (tD) y valor biológico (BV) (Miller y Bender), se determinó la calidad proteica. Se trabajó con muestras sometidas a cocción. El estudio de la concentración proteica, reveló valores similares para los tres vegetales Ks 25.8, Cha 25.0, Bv 22.1 g/100 g. Del análisis de ácidos grasos se observó un alto porcentaje de ácidos grasos insaturados correspondiendo el mayor 82 por ciento a Kochia scoparia, con predominio de ácido linolénico. Al trabajarse con material cocido los tenores de antinutrientes pueden considerarse en niveles que no comprometen la salud. Con respecto a la calidad biológica se obtuvieron los siguientes datos para Ks, Cha, Bv: NPU 68.0 + 0.4, 55.0 + 6.1, 56.0 + 4.2; tD 70.0 + 0.7, 71.0 + 4.0, 76.0 + 8.2; BV 97, 77 y 74 respectivamente. Del análisis de los indicadores por los cuales se evaluó la calidad biológica se encontró un mayor aprovechamiento nitrogenado en Ks. Por lo que Kochia scoparia cumpliría el objetivo previamente propuesto.

Processing of quinoa (Chenopodium quinoa, Willd): effects on in vitro iron availability and phytate hydrolysis.

The effect of different processing techniques was studied on in vitro iron availability and phytate hydrolysis in high and low saponin content quinoa (Chenopodium quinoa, Willd) seeds. Water slurries of ungerminated and germinated quinoa flour were processed by cooking, soaking, and fermentation using Lactobacillus plantarum as starter. Iron solubility under physiological conditions (in vitro) was measured and used as an estimation of iron availability. Phytate (inositol hexaphosphate/IP6) and its degradation products were analysed by an HPLC method. The IP6 + IP5 content was reduced by cooking with 4 to 8%, germination with 35 to 39%, soaking with 61 to 76% and by fermentation with 82 to 98%. The highest reduction, about 98%, was obtained after fermentation of the germinated flour. Cooking had no effect on the amount of soluble iron. Iron solubility increased, however, two to four times after soaking and germination, three to five times after fermentation and five to eight times after fermentation of the germinated flour samples and was highly correlated to the reduction of IP6 + IP5 (P < 0.001). There was no difference between the quinoa varieties with regard to phytate reduction and iron solubility. The pH in fermented samples was reduced from 6.5 to about 3.5, due to lactic acid formation.

Determination of provitamin A of green leafy vegetables by high performance liquid chromatography and open column chromatography.

The purpose of study was to determine the provitamin A value through beta-carotene analysis of five Brazilian leafy vegetables: carrot leaves (Daucus carota L.), beet leaves (Beta vulgaris L.) "serralha" (Sonchus oleracea L.), mint (Mentha piperita LL.) and celery leaves (Aplum graveolens. L.). Two analysis methods were used: open-column chromatography (OCC) and high performance liquid chromatography (HPLC). Two analysis methods were used: open-column chromatography (OCC) and high performance liquid chromatography (HPLC). Recovery tests were done for both methods and 92% of recovery was obtained for OCC and for HPLC 102%. The provitamin A value obtained for these leaves, using OCC were: 491 RE/100 g for carrot, 1097 for beet, 1390 for "serralha", 1016 for mint and 911 for celery. The results obtained using HPLC were 495 RE/100 g for carrot leaves, 1095 for beet leaves, 1472 for "serralha" leaves, 940 for mint leaves and 908 for celery leaves. We concluded that both methods presented good recoveries, are similar for determination of provitamin A in the leaves and they can be used for routine analysis. These five leaves can be as an inexpensive and easily obtained sources of provitamin A in Brazil.

Biological and chemical study of paico (Chenopodium chilense, Chenopodiaceae).

The methanolic extract of the aerial portion of Chenopodium chilense Schrad., used in Chilean traditional medicine as a remedy for stomach-ache, has been found to exert the major spasmolytic activity in acetylcholine contracted rat ileum. This extract, with a complex flavonoid patterns on thin layer chromatography (TLC) analysis, is practically non-toxic both for rats and brine shrimp Artemia salina in acute toxicity test.