Anti-Inflammatory Properties and Identification of Putative Protein Targets of Phytochemicals Present in Mashua (Tropaeolum Tuberosum) Tuber and Acerola (Malpighia Emarginata) Leaf by DARTS Methodology

Cellular targets for the majority of dietary phytochemicals remain unknown, presenting a challenge for the understanding of how these dietary components provide health benefits. Recently a new methodology, drug affinity responsive target stability (DARTS), was developed for this purpose. The objective of this study was to evaluate the anti-inflammatory properties of mashua (Tropaeolum tuberosum) tuber and acerola leaf (Malpighia emarginata) extracts and to identify the protein targets of its phytochemicals by using DARTS methodology. The first aim was to investigate the anti-inflammatory properties of mashua extracts and to identify the cellular targets of its phytochemicals. Mashua 203040 and 203081 extracts exerted anti-inflammatory properties in LPS-stimulated macrophages. Through DARTS methodology we identified pyruvate kinase M (PKM) as a potential target of the phytochemicals present in both extracts. Additionally, mashua extracts reduced reactive oxygen species (ROS) levels and consequently affected the expression of ROS dependent LPS-induced pro-inflammatory genes. LC-MS profiling revealed that both mashua extracts contained mainly phenolics and isothiocyantes. In this study, we demonstrated that phytochemicals from mashua reduce LPS-induced inflammation in RAW264.7 macrophages by targeting PKM and reducing ROS. The second aim was to identify the bioactive compounds acting as COX-2 inhibitors in acerola leaf by using bioassay-guided fractionation and DARTS methodology. The leaf extract (F4) was fractionated into three fractions (F1, F2, and F3) and results indicated that F4, F2, and F3 inhibited COX-2 activity. F2 showed the highest COX-2 inhibitory effect among the fractions, and also showed binding to COX-2 by DARTS. Thus, its subfractions (F2A, F2B, and F2C) were further tested for their ability to bind and inhibit COX-2 activity. All subfractions inhibited COX-2 activity but only F2A and F2C showed binding by DARTS. F2C was further fractionated and only fraction F2C4, containing terpenes, inhibited COX-2 activity. In this study, we identify the group of compounds from acerola leaf, which are responsible for the inhibition of COX-2 activity. In conclusion we were able to characterize many of the phytochemicals present in mashua tuber and acerola leaf and to identify the putative targets responsible for their anti-inflammatory effect in vitro

Suppression of oxidative stress by grape seed supplementation in rats

Polyphenol-rich grape seeds have a beneficial effect on human health. The present study was performed to investigate the effects of grape seeds on antioxidant activities in rats. Male Sprague-Dawley rats were randomly divided into a control diet group (C), a high-fat diet group (HF), a 5% grape seed-supplemented control diet group (G), and a 5% grape seed-supplemented high-fat diet group (HG). Dietary supplementation with grape seeds reduced serum concentrations of lipid peroxides compared with those in the C and HF groups. The hepatic level of lipid peroxides decreased significantly in the grape seed groups compared with that in the C and HF groups. Superoxide dismutase activity in the G group increased significantly compared with that in the C group. Catalase activity tended to be higher by feeding grape seeds. The grape seed diet increased glutathione peroxidase activity in the C group. Glutathione-S-transferase activity increased significantly in the G group compared with that in the C group. Hepatic content of total glutathione increased significantly in the HG group but decreased significantly in the HF group. The ratio of reduced glutathione and oxidized glutathione increased by feeding the grape seed diet. Total vitamin A concentration was significantly higher in HG group than in other groups. Liver tocopherol content of the G and HG groups was significantly higher than that of the control groups. These results suggest that dietary supplementation with grape seeds is beneficial for suppressing lipid peroxidation in high fat-fed rats

A simple dummy liver assist device prolongs anhepatic survival in a porcine model of total hepatectomy by slight hypothermia

<p>Abstract</p> <p>Background</p> <p>Advances in intensive care support such as therapeutic hypothermia or new liver assist devices have been the mainstay of treatment attempting to bridge the gap from acute liver failure to liver transplantation, but the efficacy of the available devices in reducing mortality has been questioned. To address this issue, the present animal study was aimed to analyze the pure clinical effects of a simple extracorporeal dummy device in an anhepatic porcine model of acute liver failure.</p> <p>Methods</p> <p>Total hepatectomy was performed in ten female pigs followed by standardized intensive care support until death. Five animals (dummy group, n = 5) underwent additional cyclic connection to an extracorporeal dummy device which consisted of a plasma separation unit. The separated undetoxified plasma was completely returned to the pigs circulation without any plasma substitution or exchange in contrast to animals receiving intensive care support alone (control group, n = 5). All physiological parameters such as vital and ventilation parameters were monitored electronically; laboratory values and endotoxin levels were measured every 8 hours.</p> <p>Results</p> <p>Survival of the dummy device group was 74 ± 6 hours in contrast to 53 ± 5 hours of the control group which was statistically significant (p < 0.05). Body temperature 24 hours after hepatectomy was significantly lower (36.5 ± 0.5°C vs. 38.2 ± 0.7°C) in the dummy device group. Significant lower values were measured for blood lactate (1.9 ± 0.2 vs. 2.5 ± 0.5 mM/L) from 16 hours, creatinine (1.5 ± 0.2 vs. 2.0 ± 0.3 mg/dL) from 40 hours and ammonia (273 ± 122 vs. 1345 ± 700 μg/dL) from 48 hours after hepatectomy until death. A significant rise of endotoxin levels indicated the onset of sepsis at time of death in 60% (3/5) of the dummy device group animals surviving beyond 60 hours from hepatectomy.</p> <p>Conclusions</p> <p>Episodes of slight hypothermia induced by cyclic connection to the extracorporeal dummy device produced a significant survival benefit of more than 20 hours through organ protection and hemodynamic stabilisation. Animal studies which focus on a survival benefit generated by liver assist devices should especially address the aspect of slight transient hypothermia by extracorporeal cooling.</p

ICAR: endoscopic skull‐base surgery

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Phytochemical characterization, antioxidant capacity and in vitro inhibition of adipogenesis and inflammation of phenolic extracts in Vaccinium floribundum and Aristotelia chilensis

Interest in berries from South America has increased due to their potential health benefits. The objective of this study was to characterize the anthocyanins and proanthocyanidins total phenolics and antioxidant capacity (AC) of V. floribundum and A. chilensis, and evaluate, in vitro, the ability of their phenolic extracts to reduce adipogenesis, and lipid accumulation in 3T3-L1 adipocytes. The anti-inflammatory properties of these extracts on RAW 264.7 macrophages was also investigated. The berry of A. chilensis contained 45.7 mg/g DW (cyanidin-3-glucososide (C3G) equivalents) total anthocyanins. Seven main anthocyanin structures were identified in this berry of which delphinidin-3-glucoside was the main anthocyanin component in this berry. The berry and the commercial powder of V. floribundum contained 10.6 mg/g DW and 2.4 mg/g (C3G equivalents) respectively of total anthocyanins. Five main anthocyanin structures where identified of which delphinidin-3- arabinose and cyanidin-3-arabinose were the main anthocyanins present in the berry and in the commercial powder of V. floribundum. The berry of A. chilensis contained 4.0 mg/g DW (epicatechin equivalents) of total proanthocyanidins, while the berries and commercial powder from V. floribundum had a slightly higher concentration of 5.2 and 4.8 mg/g DW (epicatechin equivalents) respectively. A. chilensis contained mainly proanthocyanidin dimers (56%) and trimers (14%). V. floribundum contained trimers (68%) and in less proportion pentamers (16%) and hexamers (8%). The commercial powder contained a larger percentage of dimers (34%) and trimers (23%). The berries of A. chilensis and V. floribundum showed similar values of total phenolics; however, AC was higher in the berries of A. chilensis. The commercial powder of V. floribundum had a lower phenolic content and AC than the freeze dried berry. Total phenolics and AC were highly correlated for A. chilensis and both the berries and the commercial powder of V. floribundum with R2 values of 0.90, 0.86 and 0.78, respectively. Anthocyanins were more highly correlated with AC than proanthocyanins. Phenolic extracts of the two berries and the commercial powder inhibited lipid accumulation by 4.0 to 10.8% when adipocytes were treated at maturity and by 5.9 to 37.9% when treated throughout differentiation. Furthermore, a proanthocyanidin-enriched-fraction from V. floribundum significantly induced Pref-1 expression in preadipocytes. Phenolic extracts decreased the production of nitric oxide (3.7 - 25.5%) and prostaglandin E2 (9.1 - 89.1%) and the expression of inducible nitric oxide synthase (9.8 - 61.8 %) and cycloxygenase-2 (16.6 - 62.0%) in lipopolysaccharide-stimulated RAW 264.7 macrophages. V. floribundum and A. chilensis phytochemicals limit adipogenesis and inflammatory pathways in vitro, warranting further in vivo studies

Kurz- und langfristige Sukzession in intermittierenden Flüssen : Struktur mikrobieller Sedimentgemeinschaften und biogeochemische Trajektorien

Intermittent rivers and ephemeral streams (IRES) experience drying events, and they are becoming increasingly common due to climate change and anthropogenic water withdrawal. Despite their prevalence, IRES have been less studied than non-drying rivers, and more research is needed to understand their dynamics. This dissertation examines drying events' effects on the sediment microbial community structure and biogeochemistry in temperate IRES. First, a research study was carried out using an outdoor mesocosm setup to track changes in bacterial and fungal communities, microbial functions and properties of sediment-dissolved organic carbon during three different drying events. Results showed that drying duration and intensity and sediment organic matter content regulated river Spree sediment microbial community composition and biogeochemical processes during drying. The associated sediment respiration, assessed as CO2 flux, rapidly declined during all drying treatments, and Bacilli class became the most dominant bacteria after 90 days of drying, except in non-enriched sediment under the most intense drying. Second, a microcosm experiment was conducted to understand how sediment drying history and properties influence biogeochemical dynamics during an IRES expansion phase. Sediment respiration and nutrient release/retention rates were measured through flow resumption, and results showed that drying duration and intensity and sediment organic matter content regulated the dynamics of nutrients and respiration upon flow resumption. Under the most intense and prolonged drying, non-enriched sediment showed a lag response in respiration during the first day after flow resumption, while all other treatments had either a linear increase or a fast pulse in respiration. Finally, the long-term microbial succession in IRES sediment and soil in the experimental catchment of Chicken Creek was investigated. In 2008 and 2018, the sediment of three ephemeral streams and their adjacent soil was analyzed for the structure of the bacterial and fungal communities and microbial functions across different seasons. Results showed that environmental factors, such as vegetation type and organic matter content, mediated sediment and soil microbial succession in Chicken Creek. Ephemeral streams under spare and sporadic flow events remained unique ecosystems with distinctive microbial communities and biogeochemical dynamics, different from its riparian soil. Overall, this dissertation concludes that temperate IRES are dynamic ecosystems where dry-wet cycles and environmental factors modulate their sediment microbial community structure and function trajectories in the short- and long term.Periodisch wiederkehrende Bäche und Flüsse (IRES, intermittent rivers and ephemeral streams) sind Gewässerkörper die episodisch austrocknen und durch Klimawandel und anthropogener Wasserentnahme zunehmend häufiger werden. Obwohl sie weit verbreitet sind, sind IRES bisher weit weniger erforscht worden als dauerhaft wasserführende Flüsse. Die vorliegende Dissertation untersucht Auswirkungen von Trockenperioden in IRES der gemäßigten Klimazone auf biogeochemische Parameter des Sediments sowie bakterielle und pilzliche Gemeinschaften. Zum einen wurde in einem Mesokosmen-Experiment im Freiland die bakterielle und pilzliche Diversität, sowie Eigenschaften von sedimentären, gelösten organischem Kohlenstoff und mikrobiellen Funktionen für verschiedene Austrocknungsintensitäten untersucht. Die Dauer und Intensität der Austrocknung, sowie die Konzentration an organischer Substanz, bewirkten maßgeblich eine Veränderungen der Struktur der mikrobiellen Gemeinschaft sowie der biogeochemischen Prozesse im Spree-Sediment während der Trockenphasen. In allen Austrocknungsintensitäten nahm die Respiration aus dem Sediment, gemessen als CO2 Emissionen, schnell ab; wobei die Ordnung Bacilli nach 90 Tagen Trockenzeit dominant wurde. Ausnahme war hier Sediment das nicht mit organischer Substanz angereichert war. Zum anderen wurde in Mikrokosmen der biogeochemische Effekte von vergangenen Austrocknungsepisoden und deren Eigenschaften während der Erweiterungsphase der IRES untersucht. Respiration im Sediment, sowie Freisetzung und Rückhalt von Nährstoffen wurden nach wieder einsetzenden des Wasserflusses gemessen. Eigenschaften wie Dauer und Intensität der Austrocknungsepisoden, sowie der Gehalt an organischer Substanz, bestimmten hier Nährstoffflüsse und Respiration; speziell in den ersten Tagen nach wiederkehren des Wasserflusses. Bei sehr intensiver und andauernden Austrocknung zeigte das Sediment ohne Anreicherung mit organischer Substanz eine verzögerte Reaktionszeit in der Respiration. Letzteren wurde die langfristige Auswirkung und mikrobielle Sukzession von IRES auf Sedimente und Böden im Einzugsgebiet Hühnerwasser in der Jahren 2008 und 2018 untersucht. Analysiert wurden die Struktur der mikrobiellen Gemeinschaften und Funktionen aus Sediment der wiederkehrenden Bäche und angrenzenden Uferböden in beiden Jahren. Vegetationsform und organische Substanz regulierten die mikrobielle Sukzession im Einzugsgebiet des Hühnerwassers. Die IRES bleiben auch bei nur geringem Wasserfluss eigenständige Systeme, die sich auch hinsichtlich ihrer mikrobiellen Struktur und Biogeochemie von den Böden in Gewässernähe unterscheiden. Schlussfolgernd für diese Dissertation steht, dass IRES in der temperten Klimazone dynamische Ökosysteme sind. Besonders Nass-Trocken-Zyklen und Umweltfaktoren regulieren die Struktur der mikrobiellen Gemeinschaften im Sediment und ihre Funktionen auf kurz- und langfristige Sicht

Genotipificación de cepas de Eschreichia coli enteropatogénica aisladas en una región rural de la de la costa ecuatoriana mediante el uso de la electroforesis de campo pulsado

The enteropathogenic Escherichia coli is an important category of diarrheagenic E. coli which has been linked to infant diarrhea in the developing world. Pulsed-Field Gel Electrophoresis (PFGE) is a technique used in molecular epidemiology for genotyping many pathogenic bacteria. PFGE was used to analyze 8 EPEC strains isolated from different communities in the north of Esmeraldas, Ecuador. The aim of this study was to determine the clonality of EPEC strains isolated from the different communities in the area.La Escherichia coli enteropatogénica corresponde a una categoría importante dentro de las cepas de E. coli que causan diarrea y se la ha relacionado principalmente con los casos de diarrea en niños en los países en vías de desarrollo. La electroforesis de campo pulsado (PFGE) es una técnica utilizada en la epidemiologia molecular para la genotipificación de muchas bacterias patógenas. En este estudio se utilizó el PFGE para analizar 8 cepas de EPEC aisladas de distintas comunidades del norte de la provincia de Esmeraldas