Properties of cell wall polysaccharides of raw nectarine fruits after treatment under conditions that modulate gastric digestion.

The results of the study of the physicochemical properties of the high-molecular-weight soluble and insoluble components of nectarine cell walls obtained by fruit treatment under conditions that modulate of gastric digestion are presented. Homogenized nectarine fruits were sequentially treated by natural saliva and simulated gastric fluid (SGF) at pH 1.8 and 3.0. The isolated polysaccharides were compared with polysaccharides obtained by sequential extraction of nectarine fruit with cold, hot, and acidified water, solutions of ammonium oxalate and sodium carbonate. As a result, high-molecular-weight water-soluble pectic polysaccharides, weakly bound in the cell wall, were dissolved in the simulated gastric fluid, regardless of pH. Homogalacturonan (HG) and rhamnogalacturonan-I (RG-I) were identified in all pectins. It was shown that their quantity and ability to form highly viscous solutions determine high values of the rheological characteristics of the nectarine mixture formed under simulated gastric conditions. The modifications occurring with the insoluble components under the influence of acidity of SGF were importance. They determined difference in the physicochemical properties of both the insoluble fibres and the nectarine mixtures.

Mitochondria metabonomics of Huangqi Jianzhong Tang against chronic atrophic gastritis.

Huangqi Jianzhong Tang (HQJZ) is a representative prescription used for clinical treatment of chronic atrophic gastritis (CAG) in Chinese medicine. Our previous study had revealed that energy regulation was one of the important mechanisms of HQJZ action against CAG. In this study, ultra-high-performance liquid chromatography coupled with quadrupole-Exactive mass spectrometry (UHPLC-Q-Exactive MS) based metabonomics was used to find the potential mitochondrial biomarkers and metabolic pathways of HQJZ in CAG rats, which focused on a specific organelle (mitochondria) isolated from gastric tissue samples. A total of 16 biomarkers from CAG tissues were identified with 11 of these significantly regulated by HQJZ treatment. These biomarkers was mainly involved in glycine, serine, and threonine metabolism; aminoacyl-tRNA biosynthesis metabolism; and taurine and hypotaurine metabolism. Our results show that HQJZ could protect from CAG by altering the mitochondrial function. These findings deepen our understanding of the mitochondrial metabolic changes that occur with CAG and shine a light on the mechanism of HQJZ.

The Efficacy of Processing Strategies on the Gastroprotective Potentiality of Chenopodium quinoa Seeds.

The current study has been conducted to evaluate the effect of different processing techniques on the 2,2-diphenyl-1-picrylhydrazyl (DPPH) scavenging capacity and the gastroprotective potential of Chenopodium quinoa red seeds in acute gastric injury induced by absolute ethanol in rats. Seven groups of female Sprague Dawley rats were assigned to normal and absolute ethanol (absolute EtOH) groups, given distilled water, reference control omeprazole (OMP, 20 mg/kg), pressure-cooked quinoa seeds (QP, 200 mg/kg), first stage-germinated quinoa seeds (QG, 200 mg/kg), Lactobacillus plantarum bacteria-fermented quinoa seeds (QB, 200 mg/kg), and Rhizopus oligosporus fungus-fermented quinoa seeds (QF, 200 mg/kg). One hour after treatment, all groups were given absolute ethanol, except for the normal control rats. All animals were sacrificed after an additional hour, and the stomach tissues were examined for histopathology of hematoxylin and eosin staining, immunohistochemistry of cyclooxygenase 2 (COX-2), and nitric oxide synthase (iNOS). Stomach homogenates were evaluated for oxidative stress parameters and prostaglandin E2 (PGE2). Gene expression was performed for gastric tumor necrosis factor alpha (TNF-α) and nuclear factor kappa of B cells (NF-kB). QB and QG recorded the highest DPPH scavengers compared to QF and QP. The gastroprotective potential of QB was comparable to that of OMP, followed by QF, then QG, and QP as confirmed by the histopathology, immunohistochemistry, and gene expression assessments. In conclusion, differently processed red quinoa seeds revealed variable antioxidant capacity and gastroprotective potential, while the bacterial fermented seeds (QB) showed the highest potential compared to the other processing techniques. These results might offer promising new therapy in the treatment of acute gastric injury.

The Dispersion of Diaspores of Protium icicariba (Burseraceae) - a Networked or Multifactorial System?

The adaptive radiation of the angiosperms was strongly affected by fruit and seed dispersal since the establishment of the seedlings is a fundamental process for the recruitment of juveniles to the populations. Among the species of Burseraceae, seeds with fleshy attachments and high caloric value suggest mammaliochory as an ancestral dispersal way. In Protium icicariba, at the same time as there is a visual pattern typical of ornithochory, with a report of effective demonstration, the diaspores present the highest levels of essential oils of the whole plant, suggesting other dispersion processes by olfactory guided vectors. This work aims to monitor the diasporic dispersal process in P. icicariba in situ, aiming to identify dispersers and to investigate the role of the essential oil in the dispersion of diaspores of this plant species. The natural dispersion was monitored in situ, in weekly campaigns throughout eight months, using visual and photographic records, in daily shifts of six hours, distributed along the dawn, morning, afternoon, dusk, and night. We used both direct observation and continuous picture capturing along 43 days with photographic traps. Mature diaspores removed from pseudocapsules were pooled to determine potential dispersers. Artificial models of the diaspores, in white and green colors, were also used to test hypotheses on the role of scent in the dispersion, added 1%, weight/weight, of the essential oil extracted from the mature diaspores, which chemical composition determined by gas chromatography coupled to mass spectrometry. Besides, the analysis of stomach contents of lizards collected in adjacent area was also performed. In daytime and nighttime monitoring in nature, no vertebrates were recorded dispersing diaspores. The most common was the primary wind-facilitated autochory of diaspores to the substrate, near the plant matrices. Secondarily, workers of the ant species Atta robusta can remove the pseudoarils or move the pyrenes to the anthills. The lizard species Tropidurus torquatus ingests pyrenes with the pseudoarils, and the sclerified pericarp of the pyrene is potentially resistant to chemical action of the digestive juices. Ants and lizards have also accessed the caves with natural diaspores. Concerning the artificial diaspore models, ants accessed, indistinctly, white and the green models that contained essential oils. The lizards accessed the white models, with or without essential oils, and showed insignificant access to green ones, with or without essential oil. The ingestion of pyrenes by lizards was also confirmed through analysis of stomach contents. The aggregate spatial pattern of P. icicariba at the study site, associated with clumps, may be derived from germination in the substrate near the matrices, or in the anthills or after diaspora defecation and / or regurgitation of the lizard, which is a species strongly associated with clumps of this vegetation. As the access to the diaspores by ants and lizards depends on the primary autochory, and no impediments to the germination near to the matrix plant were found, the dispersion is compatible with a multifactorial characteristic of the diplochory.

Hydrogen sulfide-mediated resistance against water avoidance stress-induced gastritis by maintenance of gastric microbial homeostasis.

Chronic persistent stress is an important cause of gastritis, but the underlying mechanism remains to be further researched, especially the role of the gastric microbiota in this process. Here, we used the water avoidance stress (WAS) test in mouse models for chronic stress-induced gastritis to investigate the underlying mechanisms of this disease. The effect of stress on the gastric microbiota was analyzed based on 16S rRNA sequencing; the changes in hydrogen sulfide (H2 S) and inflammatory cytokine levels in gastric tissues were detected by Western blotting, ELISA, immunofluorescence, and qRT-PCR. Hematoxylin and eosin staining was used as an indicator of the gastritis histological score. This finding is consistent with previous studies showing that gastric H2 S is negatively associated with the inflammatory index and might protect the gastrointestinal tract from inflammation. WAS-induced gastritis was associated with a reduction in H2 S release, which appeared to affect the homeostasis of the gastric microbiota of mice. Inflammation and microbial dysbiosis were partially reversed by sodium hydrosulfide (NaHS) and vitamin B6 (VB6) supplementation, suggesting the therapeutic potential of VB6 supplementation for the treatment of stress-induced gastritis. Gastritis has a serious impact on health and quality of life. An increasing number of people are suffering from chronic gastritis linked to a high-stress lifestyle, and our research provides clues for the prevention and treatment of stress-induced gastritis.

Microplastics in the crustaceans Nephrops norvegicus and Aristeus antennatus: Flagship species for deep-sea environments?

Ingestion of microplastics (MPs) has been documented in several marine organisms, but their occurrence in deep-sea species remains almost unknown. In this study, MPs were investigated in two economically and ecologically key crustaceans of the Mediterranean Sea, the Norwegian lobster Nephrops norvegicus and the shrimp Aristeus antennatus. Both the species were collected from 14 sites around Sardinia Island, at depths comprised between 270 and 660 m. A total of 89 and 63 stomachs were analysed for N. norvegicus and A. antennatus respectively, and more than 2000 MPs-like particles were extracted and sorted for identification and characterization by µFT-IR. In N. norvegicus, 83% of the specimens contained MPs, with an average abundance of 5.5 ±â€¯0.8 MPs individual-1, while A. antennatus showed a lower frequency of ingestion (67%) and a lower mean number of MPs (1.66 ±â€¯0.1 MPs individual-1). Composition and size of particles differed significantly between the two species. The non-selective feeding strategy of N. norvegicus could explain the 3-5 folds higher numbers of MPs in its stomach, which were mostly composed of films and fragments derived by polyethylene and polypropylene single-use plastic items. Contrarily, most MPs in the stomachs of A. antennatus were polyester filaments. The MPs abundance observed in N. norvegicus is among the highest detected in Mediterranean species considering both fish and invertebrates species, and provides novel insights on MPs bioavailability in deep-sea habitats. The overall results suggest that both N. norvegicus and A. antennatus, easily available in common fishery markets, could be valuable bioindicators and flagship species for plastic contamination in the deep-sea.

O Complexo Regulador em Saúde do Distrito Federal, Brasil, e o desafio da integração entre os níveis assistenciais / The Regulatory Complex for Health Care in the Federal District, Brazil and the challenge for integrating levels of health care

Resumo A integração entre os níveis de atenção é um desafio contínuo mesmo em sistemas de saúde consolidados e de alta performance. A reestruturação do sistema público de saúde do Distrito Federal, baseado no fortalecimento da Atenção Primária à Saúde (APS), na reconfiguração da atenção especializada (ambulatorial e hospitalar) e de sua rede de urgência e emergência, trouxe como desafio a necessidade de integração entre esses níveis. Fez-se necessária a criação de um dispositivo que desempenhasse o papel de "gatekeeper" conduzindo o acesso equânime, transparente e seguro para a atenção especializada e hospitalar. Neste sentido, foi estruturado o Complexo Regulador em Saúde do Distrito Federal (CRDF) e suas Centrais de Regulação (CR), orientados para a execução de um processo regulatório de acesso para os serviços de internação hospitalar, ambulatorial (procedimentos e consultas especializadas), cirurgias eletivas, alta complexidade, transporte sanitário, urgências e transplantes de pacientes do Distrito Federal e de fora dele. Este artigo descreve o processo de implantação e de implementação do CRDF e de suas CR, de forma a refletir sobre as potencialidades e desafios de seu papel enquanto instrumento de integração entre os níveis assistenciais de saúde.

Abstract The integration among levels of care is a continuous challenge even in consolidated and high performance health systems. The reform of public health system of Distrito Federal, based on the strengthening of primary health care associated to the reconfiguration of specialized attention (ambulatory and hospital) and of its of urgency and emergence network brought, as a challenge, the need for integration between these levels. Thus, became necessary to create an instrumentto perform the role of gatekeeper, leading to equanimous, transparent and safe access to specialized and hospital care. Thus, the Regulatory Complex in Health of Distrito Federal (CRDF) and its Regulatory Centers (CR) were created to carry out the regulatory process of access to care services, such as hospitalization, ambulatory care (procedures and specialized consultations), elective surgeries, complex procedures, sanitary transport, urgencies and transplants of patients of the Federal District and outside it. This article describes the process of the CRDF implementation and its CRs, aiming to reflect on the potential and challenges of its role as an instrument of integration among the levels of care.

Pea Ferritin Stability under Gastric pH Conditions Determines the Mechanism of Iron Uptake in Caco-2 Cells.

Background: Iron deficiency is an enduring global health problem that requires new remedial approaches. Iron absorption from soybean-derived ferritin, an ∼550-kDa iron storage protein, is comparable to bioavailable ferrous sulfate (FeSO4). However, the absorption of ferritin is reported to involve an endocytic mechanism, independent of divalent metal ion transporter 1 (DMT-1), the transporter for nonheme iron. Objective: Our overall aim was to examine the potential of purified ferritin from peas (Pisum sativum) as a food supplement by measuring its stability under gastric pH treatment and the mechanisms of iron uptake into Caco-2 cells. Methods: Caco-2 cells were treated with native or gastric pH-treated pea ferritin in combination with dietary modulators of nonheme iron uptake, small interfering RNA targeting DMT-1, or chemical inhibitors of endocytosis. Cellular ferritin formation, a surrogate measure of iron uptake, and internalization of pea ferritin with the use of specific antibodies were measured. The production of reactive oxygen species (ROS) in response to equimolar concentrations of native pea ferritin and FeSO4 was also compared. Results: Pea ferritin exposed to gastric pH treatment was degraded, and the released iron was transported into Caco-2 cells by DMT-1. Inhibitors of DMT-1 and nonheme iron absorption reduced iron uptake by 26-40%. Conversely, in the absence of gastric pH treatment, the iron uptake of native pea ferritin was unaffected by inhibitors of nonheme iron absorption, and the protein was observed to be internalized in Caco-2 cells. Chlorpromazine (clathrin-mediated endocytosis inhibitor) reduced the native pea ferritin content within cells by ∼30%, which confirmed that the native pea ferritin was transported into cells via a clathrin-mediated endocytic pathway. In addition, 60% less ROS production resulted from native pea ferritin in comparison to FeSO4. Conclusion: With consideration that nonheme dietary inhibitors display no effect on iron uptake and the low oxidative potential relative to FeSO4, intact pea ferritin appears to be a promising iron supplement.

Superdosing phytase reduces real-time gastric pH in broilers and weaned piglets.

1. The current study was conducted to investigate the effect of high phytase doses on growth performance and real-time gastric pH measurements in broiler chickens and pigs. 2. In the first experiment, 576 male Ross 308 broilers were fed in two phases (0-21 and 21-42 d) with 4 treatment groups, with diets meeting nutrient requirements containing 0, 500, 1500 or 2500 FTU/kg phytase. In the second, 64 Landrace weaners were fed on diets meeting nutrient requirements with or without phytase (0 or 2500 FTU/kg) in two phases (0-21 and 21-42 d). Heidelberg pH capsules were administered to 7 broilers and approximately 13 pigs per treatment group, pre- and post-phase change, with readings monitored over several hours. 3. Addition of phytase into an adequate Ca and P diet had no significant effect on broiler performance although phytase tended (P < 0.07) to improve feed conversion in pigs over the entire experimental period. Real-time pH capsule readings in broilers demonstrated an increase (P < 0.05) in gizzard pH when phytase was dosed at 500 or 1500 FTU/kg, while higher doses of 2500 FTU/kg phytase lowered pH to a level comparable to control birds. Gastric pH increased (P < 0.01) when animals were exposed to dietary phase change, signifying a potential challenge period for nutrient digestibility. However, pigs fed 2500 FTU/kg were able to maintain gastric pH levels through diet phase change. In contrast, spear-tip probe measurements showed no treatment effect on gastric pH. 4. These findings demonstrate dietary manipulation of gastric pH and the value of real-time pH capsule technology as a means of determining phytase dose response.

Seeds of Buchholzia coriacea in Diet Mitigate Ischemic Reperfusion-Induced Gastric Ulceration in Experimental Rats.

Buchholzia coriacea (B. coriacea) seeds, in folk medicine, have been documented to prevent gastric ulceration though the mechanism is not fully elucidated. To clarify this, the gastro-healing activities were investigated using graded incorporation of B. coriacea seeds in the diet. Male Wistar rats (150-200 g) were divided into 7 groups (n = 15): unulcerated untreated control, ulcerated untreated control, unulcerated B. coriacea low (10%), ulcerated B. coriacea low (10%), nulcerated B. coriacea high (25%), ulcerated B. coriacea high (25%), and ulcerated omeprazole-treated groups. Rats were fed with B. coriacea diets for 7 weeks; thereafter, ulcer was induced by ischemic reperfusion method. Daily body weight, gastric acid secretion, hematological parameters, stomach ulcer score, and biochemical and histological analyses were evaluated on days 0, 3, and 7 post-ulcer induction. Results were subjected to one-way analysis of variance (ANOVA) and presented as mean ± standard error of the mean (SEM); p ≤.05 was considered significant. Significant decreases were observed in mean body weight of B. coriacea-fed compared with control and omeprazole-treated groups from week 7. Ulcerated B. coriacea-fed showed significant decrease in gastric acid secretion by days 3 and 7 compared with ulcerated control groups. Malondialdehyde content was significantly decreased in ulcerated B. coriacea-fed compared with control and omeprazole-treated groups. Significant increases in hematological variables (notably platelet count), superoxide dismutase, catalase, and nitric oxide levels of B. coriacea-fed compared with control and omeprazole-treated groups by days 0 and 3 were observed. Histological evaluations further confirmed these observations. B. coriacea diet enhanced gastric healing activities on ischemic reperfused gastric ulcer. Increased platelet count and nitric oxide levels may play significant roles in this process.

Synergistic gastroprotective activity of methanolic extract of a mixture of Melastoma malabathricum and Muntingia calabura leaves in rats.

BACKGROUND: Melastoma malabathricum L. (family Melastomaceae; MM) and Muntingia calabura L. (family Elaeocarpaceae; MC) have been separately reported to possess gastroprotective activity. In an attempt to develop a pharmaceutical product with antiulcer potential, the synergistic gastroprotective activity of methanolic extract of a mixture of MM and MC (MMMC) at various ratios was evaluated in rat models. METHODS: Rats were pre-treated orally with 2% Tween 80 (vehicle), 100 mg/kg ranitidine (reference drug) or MMMC (ratios of 1:1, 1:3 and 3:1 (v/v); doses of 15, 150 or 300 mg/kg) and then subjected to the ethanol-induced gastric ulcer or pyloric ligation assays. Stomach of rats from the former assay was collected and subjected to the macroscopic and microscopic observations, and enzymatic and non-enzymatic antioxidant studies while the gastric juice content and tissue from the latter assay were subjected to the antisecretory activity study. The UHPLC analysis of MMMC was also performed. RESULT: MMMC, in the ratio 1:1, demonstrated the most effective (P < 0.001) gastroprotective activity indicated by the highest reduction in ethanol-induced ulcer area formation. These macroscopic findings were supported by the microscopic observations. Except for pH and total acidity, MMMC also significantly (P < 0.001) reduced the volume of gastric content but increased the gastric wall mucus content in the pyloric-ligation test. MMMC also demonstrated remarkable antioxidant activity indicated by the highest total phenolic content (TPC) value and oxygen radical absorbance capacity (ORAC) activity with the recorded IC50 value of approximately 53 µg/mL for the 2,2- diphenyl-1-picrylhydrazyl (DPPH) scavenging activity. MMMC also improved the catalase (CAT), superoxide dismutase (SOD), glutathione (GSH), prostaglandin E2 (PGE2) and malondialdehyde (MDA) activities of the gastric tissue intoxicated by ethanol. UHPLC analysis of MMMC confirmed the presence several flavonoid-based bioactive compounds. CONCLUSION: MMMC, at the ratio of 1:1 (v/v), exerts gastroprotective activity partly by activating its antisecretory and antioxidant activities, and via modulation of the gastric tissue endogenous antioxidant system.

A comparative study of the antacid effect of some commonly consumed foods for hyperacidity in an artificial stomach model.

OBJECTIVES: The incorporation of certain alkalinizing vegetables, fruits, milk and its products in the diet has been known to alleviate hyperacidity. These foods help to restore the natural gastric balance and function, curb acid reflux, aid digestion, reduce the burning sensation due to hyperacidity and soothe the inflamed mucosa of the stomach. The present study evaluates and compares the antacid effect of broccoli, kale, radish, cucumber, lemon juice, cold milk and curd in an artificial stomach model. DESIGN: The pH of the test samples and their neutralizing effect on artificial gastric acid was determined and compared with that of water, the active control sodium bicarbonate and a marketed antacid preparation ENO. A modified model of Vatier's artificial stomach was used to determine the duration of consistent neutralization of artificial gastric acid by the test samples. The neutralizing capacity of the test samples was determined in vitro using the classical titration method of Fordtran. RESULTS: All test samples except lemon showed significantly higher (p<0.05 for cucumber and p<0.001 for the rest) acid neutralizing effect than water. All test samples also exhibited a significantly (p<0.001) higher duration of consistent neutralization and higher antacid capacity than water. Highest antacid activity was demonstrated by cold milk and broccoli which was comparable with ENO and sodium bicarbonate. CONCLUSION: It may be concluded that the natural food ingredients used in this study exhibited significant antacid activity, justifying their use as essential dietary components to counter hyperacidity.

The Physical State of Emulsified Edible Oil Modulates Its in Vitro Digestion.

Emulsified lipid digestion was tailored by manipulating the physical state of dispersed oil droplets in whey protein stabilized oil-in-water (O/W) emulsions, where the oil phase consisted of one of five ratios of soybean oil (SO) and fully hydrogenated soybean oil (FHSO). The evolution in particle size distribution, structural changes during oral, gastric, and intestinal digestion, and free fatty acid release during intestinal digestion were all investigated. Irrespective of the physical state and structure of the dispersed oil/fat, all emulsions were stable against droplet size increases during oral digestion. During gastric digestion, the 50:50 SO:FHSO emulsion was more stable against physical breakdown than any other emulsion. All emulsions underwent flocculation and coalescence or partial coalescence upon intestinal digestion, with the SO emulsion being hydrolyzed the most rapidly. The melting point of all emulsions containing FHSO was above 37 °C, with the presence of solid fat within the dispersed oil droplets greatly limiting lipolysis. Fat crystal polymorph and nanoplatelet size did not play an important role in the rate and extent of lipid digestion. Free fatty acid release modeled by the Weibull distribution function showed that the rate of lipid digestion (κ) decreased with increasing solid fat content, and followed an exponential relationship (R2 = 0.95). Overall, lipid digestion was heavily altered by the physical state of the dispersed oil phase within O/W emulsions.

Fatal Folic Acid Toxicity in Humans.

Folic acid is B-9 vitamin. Folic acid is prescribed commonly for pregnant women to prevent neural tube defects in the fetus, patients under chemotherapy, pernicious anemia and to reduce the risk of stroke and cardiovascular disease. Acute or chronic ingestion of a large dose of folic acid generally manifests as neurological complications, which are reversible. In this present case, a 23-year-old pregnant woman committed suicide by consuming folic acid tablets and succumbed to death within 36 h. Postmortem toxicological analysis detected folic acid in viscera. Death following acute consumption of folic acid is rare and has been not reported in the literature, to the best of our knowledge.

A comparative study of the antacid effect of raw spinach juice and spinach extract in an artificial stomach model.

BackgroundSpinacia oleracea known as spinach is a green-leafy vegetable consumed by people across the globe. It is reported to possess potent medicinal properties by virtue of its numerous antioxidant phytoconstituents, together termed as the natural antioxidant mixture (NAO). The present study compares the antacid effect of raw spinach juice with an antioxidant-rich methanolic extract of spinach (NAOE) in an artificial stomach model. MethodsThe pH of NAOE at various concentrations (50, 100 and 200 mg/mL) and its neutralizing effect on artificial gastric acid was determined and compared with that of raw spinach juice, water, the active control sodium bicarbonate (SB) and a marketed antacid preparation ENO. A modified model of Vatier's artificial stomach was used to determine the duration of consistent neutralization of artificial gastric acid for the test compounds. The neutralizing capacity of test compounds was determined in vitro using the classical titration method of Fordtran. Results NAOE (50, 100 and 200 mg/mL), spinach juice, SB and ENO showed significantly better acid-neutralizing effect, consistent duration of neutralization and higher antacid capacity when compared with water. Highest antacid activity was demonstrated by ENO and SB followed by spinach juice and NAOE200. Spinach juice exhibited an effect comparable to NAOE (200 mg/mL). ConclusionsThus, it may be concluded that spinach displays significant antacid activity be it in the raw juice form or as an extract in methanol.

Pharmacokinetics, biodistribution and excretion studies of neotuberostemonine, a major bioactive alkaloid of Stemona tuberosa.

Neotuberostemonine is a potent antitussive alkaloid extracted from Stemona tuberosa. However, the pharmacokinetics, tissue distribution and excretion of pure neotuberostemonine have not been reported. The present study was aimed to investigate the pharmacokinetic parameters of neotuberostemonine by developing an ultra-high performance liquid chromatography-tandem mass spectrometry method. Neotuberostemonine and tetrahydropalmatine (internal standard, IS) in bio-samples were extracted by protein precipitation with methanol and successfully separated on a Zorbax Extend C18 column by using a mobile phase of acetonitrile and a mixture of 0.1% formic acid and 5mM ammonium acetate. The detection was performed by using positive ion electrospray ionization in multiple reaction monitoring mode. The MS/MS ion transitions were monitored at m/z 376.1→302.0 for neotuberostemonine and 355.8→192.0 for IS. After oral administration of neotuberostemonine in rats, the Cmax and AUC0-∞ were 11.37ng/mL and 17.68ng·h/mL at 20mg/kg and 137.6ng/mL and 167.4ng·h/mL at 40mg/kg, and the t1/2 were 2.28 and 3.04h at 20 and 40mg/kg, respectively. The high neotuberostemonine concentrations were found in intestine, stomach and liver, and there was no long-term accumulation of neotuberostemonine in tissues. Total recoveries of neotuberostemonine were only 0.90% (0.19% in bile, 0.05% in urine and 0.66% in feces), which might be resulted from the intestine and liver first-pass effects, indicating that neotuberostemonine may be mainly excreted as its metabolites. All above results would provide helpful information for the further pharmacological and clinical studies of neotuberostemonine and the crude drug.

Development of a liquid chromatography with mass spectrometry method for the determination of gelsemine in rat plasma and tissue: Application to a pharmacokinetic and tissue distribution study.

Gelsemine from Gelsemium elegans Benth is a potential anesthetic and analgesic agent with no physical dependence and opiate addiction. This study was aimed at developing an ultrafast liquid chromatography coupled to tandem mass spectrometry method to quantify gelsemine in rat plasma and tissues. Plasma and tissues were processed with acetonitrile precipitation, and dendrobine was chosen as the internal standard. Sample separation was performed on an ACQUITY HSS T3 column. The mobile phase consisted of acetonitrile and 0.1% formic acid aqueous solution. Multiple reactions monitoring mode was utilized to detect the compounds of interest. The mass spectrometer was operated in the positive ion mode for detection. The MS/MS ion transitions monitored were m/z 323.2→70.5 for gelsemine and 264.2→108.05 for dendrobine, respectively. The calibration curves were linear over the range of 1-500 ng/mL in all biological matrices. The lower limit of quantification for rats plasma and tissues was 1.0 ng/mL. The values for inter- and intraday precision and accuracy were well within the ranges acceptable (< 15%). It was successfully applied to the pharmacokinetic and tissue distribution studies of gelsemine after intravenous doses of 5, 2, and 0.5 mg/kg in rats. These data of gelsemine would be useful for clinical application and further development.

[Establishment of animal model with gastric cold syndrome].

This study is to establish the gastric cold model of rats. After gastric feeding with cold water for 5 weeks and extra iced water bath in the last 2 weeks, model group show distinct physical sign of gastric cold syndrome. The pathology of gastrics reveals gastricism of model group, while treatment group(treated with Fanzuojin Wan) show mild lesion. Elisa detection of model group show that the solution of interleukin-2 (IL-2) is higher than blank group. The difference with significance among model group, treatment group and blank group reveals the success of the establishment of gastric cold syndrome.

Olives and olive oil are sources of electrophilic fatty acid nitroalkenes.

Extra virgin olive oil (EVOO) and olives, key sources of unsaturated fatty acids in the Mediterranean diet, provide health benefits to humans. Nitric oxide (•NO) and nitrite (NO2 (-))-dependent reactions of unsaturated fatty acids yield electrophilic nitroalkene derivatives (NO2-FA) that manifest salutary pleiotropic cell signaling responses in mammals. Herein, the endogenous presence of NO2-FA in both EVOO and fresh olives was demonstrated by mass spectrometry. The electrophilic nature of these species was affirmed by the detection of significant levels of protein cysteine adducts of nitro-oleic acid (NO2-OA-cysteine) in fresh olives, especially in the peel. Further nitration of EVOO by NO2 (-) under acidic gastric digestive conditions revealed that human consumption of olive lipids will produce additional nitro-conjugated linoleic acid (NO2-cLA) and nitro-oleic acid (NO2-OA). The presence of free and protein-adducted NO2-FA in both mammalian and plant lipids further affirm a role for these species as signaling mediators. Since NO2-FA instigate adaptive anti-inflammatory gene expression and metabolic responses, these redox-derived metabolites may contribute to the cardiovascular benefits associated with the Mediterranean diet.