Immunomodulatory Effects of Flavonoids in the Prophylaxis and Treatment of Inflammatory Bowel Diseases: A Comprehensive Review.

Inflammatory Bowel Diseases (IBD) comprised of two disorders of idiopathic chronic intestinal inflammation that affect about three million people worldwide: Crohn's disease and ulcerative colitis. Nowadays, the first-line of treatment for patients with mild to moderate symptoms of IBD is comprised of corticosteroids, immunosuppressants, antibiotics, and biological agents. Unfortunately, none of these drugs are curative, and their long-term use may cause severe side effects and complications. Almost 40% of IBD patients use alternative therapies to complement the conventional one, and flavonoids are gaining attention for this purpose. The biological properties of flavonoids are well documented and their antioxidant and anti-inflammatory activities have been arousing attention in the scientific community. Flavonoids are the most widely distributed polyphenols in plants and fruits, making part of the human diet. Taking into account that all ingested flavonoids are expected to exert biological actions at the gastrointestinal level, research on the modulatory effect of these compounds in IBD is of paramount importance. This review intends to summarize, in an integrated and comprehensive form, the effect of flavonoids, both in vitro and in vivo, in the different phases of the characteristic IBD inflammatory network.

Solanum diploconos fruits: profile of bioactive compounds and in vitro antioxidant capacity of different parts of the fruit.

Solanum diploconos is an unexploited Brazilian native fruit that belongs to the same genus of important food crops, such as tomato (Solanum lycorpersicum) and potato (Solanum tuberosum). In this study, we determined, for the first time, the profile of bioactive compounds (phenolic compounds, carotenoids, ascorbic acid and tocopherols) of the freeze-dried pulp and peel of Solanum diploconos fruits, as well as of an extract obtained from the whole fruit. Additionally, the antioxidant potential of the whole fruit extract was evaluated in vitro, against reactive oxygen species (ROS) and reactive nitrogen species (RNS). Eighteen phenolic compounds were identified in the peel and pulp and 6 compounds were found in the whole fruit extract. Coumaric, ferulic and caffeic acid derivatives were revealed to be the major phenolic constituents. All-trans-ß-carotene was the major carotenoid (17-38 µg g(-1), dry basis), but all-trans-lutein and 9-cis-ß-carotene were also identified. The peel and pulp presented <2 µg per mL of tocopherols, and ascorbic acid was not detected. The whole fruit extract exhibited scavenging capacity against all tested ROS and RNS (IC50 = 14-461 µg mL(-1)) with high antioxidant efficiency against HOCl. Thus, Solanum diploconos fruits may be seen as a promising source of bioactive compounds with high antioxidant potential against the most physiologically relevant ROS and RNS.

pH sensitive silica nanotubes as rationally designed vehicles for NSAIDs delivery.

A novel pH-sensitive drug delivery system based on functionalized silica nanotubes was developed for the incorporation of non-steroidal anti-inflammatory drugs (NSAIDs), aimed at a tailored drug release in acidic conditions characteristic of inflamed tissues. Silica nanotubes (SNTs) were synthesized by a nanoporous alumina template assisted sol-gel method. Inner surfaces were physically and chemically modified to improve both the functionalization and subsequent incorporation of the drug. Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), energy dispersive spectroscopy (EDS) and transmission electron microscopy (TEM) were used to characterize the designed nanocarriers and their functionalization. To achieve the highest degree of functionalization, three types of aminosilanes were tested and calcination conditions were optimized. APTES was shown to be the most effective aminosilane regarding the functionalization of the SNTs' inner surface and an adequate calcination temperature (220°C) was found to attain mechanical stability without compromising functionalization efficiency. Finally, the incorporation of naproxen into the nanotubes was accessed by fluorescence measurements and drug release studies were performed, revealing that the electrostatic linkage ensures effective release of the drug in the acidic pH typical of inflamed cells, while maintaining the SNT-drug conjugates stable at the typical bloodstream pH.

Single interface flow system with potentiometric detection for the determination of nitrate in water and vegetables.

In this work a single interface flow system (SIFA) with potentiometric detection was for the first time implemented and applied to the determination of nitrate in waters and plant extracts. The analytical potential of the SIFA system was exploited not only to transport the sample towards detection but also to carry out, in a reproducible and automated way, the tasks associated with sample pre-treatment, namely ionic strength, pH adjustment and interfering species suppression. The advantageous aspects of combining a SIFA system with potentiometry with enhanced simplicity, ease of implementation and automation were further discussed and emphasised. The obtained results showed relative deviations lower than 5%, for both types of samples, with sampling rates of about 40h(-1). In addition, an innovative and straightforward process for constructing plastic membrane ion selective electrodes with a tubular configuration able to be coupled to flow-based analytical systems is also proposed. The developed approach, consisting of assembling the electrode inside a flow tubing connector is very simple to implement, robust, particularly adequate to be combined with flow methodologies and maintains all dynamic and analytical characteristics exhibited by previous assembling processes.

In vitro protective effect of Hypericum androsaemum extract against oxygen and nitrogen reactive species.

Hypericum androsaemum L. (Gutiferae) is a medicinal plant growing in Western Europe that has been used in traditional medicine in the prevention or treatment of liver diseases. Oxidative stress and nitrosative stress are common pathogenetic mechanisms contributing to initiation and progression of hepatic damage in several liver disorders. In the present study, an ethanol:water (4:6) extract from H. androsaemum branches and leaves were evaluated for its putative in vitro scavenging effects on 1,1-diphenyl-2-picrylhydrazil radical, on reactive oxygen species, namely HO•, O2•-, ROO•, ¹O2 and H2O2 and on reactive nitrogen species, namely •NO and ONOO⁻. The hypericum extract presented a remarkable capacity to scavenge all the tested reactive species, all the IC50 values being found at the µg/ml level. IC50 values for 1,1-diphenyl-2-picrylhydrazil, and for the reactive oxygen species O2•-, H2O2, HO• and ¹O2 were 11.3 ± 0.7, 32.7 ± 3.4, 944 ± 47, 595 ± 82, 28.3 ± 1.2 µg/ml respectively. The oxygen radical absorbance capacity value obtained for ROO• was 1.5 ± 0.1 µmol Trolox equivalents/mg extract. The IC50 values for •NO and ONOO⁻ were 2.2 ± 0.2 and 1.2 ± 0.1 µg/ml respectively. The content of total phenolics was 281 ± 2 mg of gallic acid equivalents/g of lyophilized extract. The observed antioxidant activity provides scientific support for the reported therapeutic use of H. androsaemum, though further in vitro and in vivo studies are required to ascertain the risk/benefit score at therapeutic concentrations.

Oxygen and nitrogen reactive species are effectively scavenged by Eucalyptus globulus leaf water extract.

Eucalyptus globulus Labill. (Family Myrtaceae) is a plant of Australian origin, with a reported therapeutic use in airway inflammatory diseases. Considering that reactive oxygen species (ROS) and reactive nitrogen species (RNS) have been implicated in the pathogenesis of airway inflammatory diseases such as asthma and chronic obstructive pulmonary disease, an effective scavenging activity against these reactive species may contribute for the therapeutic effect of this plant. In the present study, a water extract of E. globulus leaves was evaluated for its putative in vitro scavenging effects on ROS (HO(*), O(2)(*-), ROO(*), and H(2)O(2)) and RNS ((*)NO and ONOO(-)) and on 2,2-diphenyl-1-picrylhydrazyl radical (DPPH). Qualitative and quantitative analyses of the extract's phenolic composition were also performed. The Eucalyptus leaf water extract presented a remarkable capacity to scavenge all the reactive species tested, with all the 50% inhibitory concentrations being found at the mug/mL level. Phytochemical analysis showed the presence of polyphenols such as flavonoids (rutin and quercitrin) and phenolic acids (chlorogenic acid and ellagic acid), which may be partially responsible for the observed antioxidant activity. These observations provide further support, beyond the well-known antibacterial and antiviral activities of the Eucalyptus plant, for its reported use in traditional medicine such as in the treatment of airway inflammatory diseases, considering the important role of ROS and RNS in the inflammatory process, although further studies are needed to prove the bioavailability of the antioxidants/antibacterial compounds of the extract as well as the ability of the active compounds to reach specific tissues and to act in them.

Direct introduction of slurry samples in multi-syringe flow injection analysis: determination of potassium in plant samples.

The present work explores the slurry sampling approach for automatic, flow-based plant analysis. For this purpose, pinch valves were introduced into a multi-syringe flow injection analysis manifold to provide the repeatable aspiration of a few microliters of plant suspension before the material was further processed through the flow system. For validation of the proposed approach, the determination of potassium by flame emission spectrometry was implemented. Several parameters were studied: the concentration of plant particles in the sample suspension and the utilization of matrix modifiers. Microwave digestion was also implemented; no significant difference was found when certified reference material was analyzed with or without the in-line digestion step. The system was successfully applied to 13 samples within a concentration range of 2.5 to 100 mg g(-1). A determination frequency of 28 h(-1) was achieved and the precision was better than 4.0% (n = 12).

Protective effect of Castanea sativa and Quercus robur leaf extracts against oxygen and nitrogen reactive species.

Topical natural antioxidants are a useful strategy for the prevention of photoaging and oxidative stress mediated skin diseases. In view of this underlying principle, the screening of natural plant extracts with scavenging activity for pro-oxidant reactive species is a primary requirement for the development of new topical antioxidant formulations. In the present study, an ethanol:water (7:3) extract from Castanea sativa leaves and a ethanol:water (2:3) extract from Quercus robur leaves were evaluated for their putative in vitro scavenging effects on reactive oxygen species (ROS) namely superoxide radical (O(2)(-)), hydroxyl radical (HO()), peroxyl radical (ROO()), hydrogen peroxide (H(2)O(2)) and singlet oxygen ((1)O(2)) as well as on reactive nitrogen species (RNS) namely nitric oxide (()NO) and peroxynitrite (ONOO(-)). The extracts presented a high potency to scavenge the tested reactive species, all the IC(50)s being found at the microg/mL level. IC(50)s (mean+/-SE) for the ROS O(2)(-),HO(),H(2)O(2) and (1)O(2) were 13.6+/-1.8; 216+/-4; 410+/-8; 12.3+/-0.7 microug/mL, respectively, for C. sativa, and 11.0+/-0.5; 285+/-22; 251+/-32; 7.90+/-0.56 microg/mL, respectively, for Q. robur. The ORAC values obtained for ROO() were 1.24+/-0.13 for C. sativa and 1.09+/-0.06 for Q. robur. The IC(50)s (mean+/-SE) for ()NO and ONOO(-) were 3.10+/-0.14 and 1.49+/-0.10 microg/mL, respectively, for C. sativa and 3.13+/-0.11 and 0.95+/-0.02 microg/mL, respectively, for Q. robur. The content of total phenolics for C. sativa and Q. robur were 284+/-9 and 346+/-4 mg of gallic acid equivalents (GAE)/g of lyophilized extract respectively. The observed effects might be of relevance considering the putative interest of these extracts as topical antioxidants.

A multipumping flow system for in vitro screening of peroxynitrite scavengers.

Peroxynitrite anion is a reactive nitrogen species formed in vivo by the rapid, controlled diffusion reaction between nitric oxide and superoxide radicals. By reacting with several biological molecules, peroxynitrite may cause important cellular and tissue deleterious effects, which have been associated with many diseases. In this work, an automated flow-based procedure for the in vitro generation of peroxynitrite and subsequent screening of the scavenging activity of selected compounds is developed. This procedure involves a multipumping flow system (MPFS) and exploits the ability of compounds such as lipoic acid, dihydrolipoic acid, cysteine, reduced glutathione, oxidized glutathione, sulindac, and sulindac sulfone to inhibit the chemiluminescent reaction of luminol with peroxynitrite under physiological simulated conditions. Peroxynitrite was generated in the MPFS by the online reaction of acidified hydrogen peroxide with nitrite, followed by a subsequent stabilization by merging with a sodium hydroxide solution to rapidly quench the developing reaction. The pulsed flow and the timed synchronized insertion of sample and reagent solutions provided by the MPFS ensure the establishment of the reaction zone only inside the flow cell, thus allowing maximum chemiluminescence emission detection. The results obtained for the assayed compounds show that, with the exception of oxidized glutathione, all are highly potent scavengers of peroxynitrite at the studied concentrations.