Metabotropic Glutamate Receptor Blockade Reduces Preservation Damage in Livers from Donors after Cardiac Death

We previously demonstrated that the blockade of mGluR5 by 2-methyl-6(phenylethynyl)pyridine (MPEP) reduces both cold and warm ischemia/reperfusion injury. Here we evaluated whether MPEP reduces the hepatic preservation injury in rat livers from cardiac-death-donors (DCDs). Livers from DCD rats were isolated after an in situ warm ischemia (30 min) and preserved for 22 h at 4 °C with UW solution. Next, 10 mg/Kg MPEP or vehicle were administered 30 min before the portal clamping and added to the UW solution (3 µM). LDH released during washout was quantified. Liver samples were collected for iNOS, eNOS, NO, TNF-α, ICAM-1, caspase-3 and caspase-9 protein expression and nuclear factor-erythroid-2-related factor-2 (Nrf2) gene analysis. Lower LDH levels were detected in control grafts versus DCD groups. An increase in eNOS and NO content occurred after MPEP treatment; iNOS and TNF-α content was unchanged. ICAM-1 expression was reduced in the MPEP-treated livers as well as the levels of caspase-3 and caspase-9. Nrf2, oxidative stress-sensitive gene, was recovered to control value by MPEP. These results suggest that MPEP can be used to reclaim DCD livers subjected to an additional period of cold ischemia during hypothermic storage. MPEP protects against apoptosis and increased eNOS, whose overexpression has been previously demonstrated to be protective in hepatic ischemia/reperfusion damage.     

Development of a New Highly Selective Monoclonal Antibody against Preferentially Expressed Antigen in Melanoma (PRAME) and Identification of the Target Epitope by Bio-Layer Interferometry

Background: Monoclonal antibodies (mAbs) against cancer biomarkers are key reagents in diagnosis and therapy. One such relevant biomarker is a preferentially expressed antigen in melanoma (PRAME) that is selectively expressed in many tumors. Knowing mAb’s epitope is of utmost importance for understanding the potential activity and therapeutic prospective of the reagents. Methods: We generated a mAb against PRAME immunizing mice with PRAME fragment 161–415; the affinity of the antibody for the protein was evaluated by ELISA and SPR, and its ability to detect the protein in cells was probed by cytofluorimetry and Western blotting experiments. The antibody epitope was identified immobilizing the mAb on bio-layer interferometry (BLI) sensor chip, capturing protein fragments obtained following trypsin digestion and performing mass spectrometry analyses. Results: A mAb against PRAME with an affinity of 35 pM was obtained and characterized. Its epitope on PRAME was localized on residues 202–212, taking advantage of the low volumes and lack of fluidics underlying the BLI settings. Conclusions: The new anti-PRAME mAb recognizes the folded protein on the surface of cell membranes suggesting that the antibody’s epitope is well exposed. BLI sensor chips can be used to identify antibody epitopes.     

Open prostatectomy in benign prostatic hyperplasia: 10-year experience in Italy

This study reports the experience of 47 Italian urology units together with the urology unit at the University of Perugia concerning open surgery in the management of benign prostatic hyperplasia (BPH). Until 20-25 years ago, open surgery was the most common approach. In the late 1970s the development of endoscopes and their methodology has led to a gradual reduction in open surgery operations, which decreased rapidly with the introduction of mini-invasive endoscopic techniques. Therefore, open surgery for BPH is declining, though still performed. Skill in traditional surgery is mandatory because, until an alternative is devised, indications for open surgery still exist and cannot be ignored. The survey shows the indications and contraindications, complications and results of a 10-year nationwide experience. Guidelines for open surgery in patients with BPH have been drawn up.     

Exposure to physical movement in low back pain patients: Restricted effects of generalization.

Whether the effects of exposure to 1 movement generalize to another dissimilar movement was investigated in 37 patients with low back pain (15 men, 22 women). Two movements were executed twice: bending forward while standing and lifting 1 leg while lying down. During each trial, baseline pain, expected pain, and experienced pain were recorded. Similar ratings for perceived harm were obtained. Analyses revealed an initial overprediction of pain, but after exposure the overprediction was readily corrected. This exposure effect did not generalize toward another dissimilar movement. These results were only characteristic for patients with catastrophic thinking about pain. Low pain catastrophizers did not overpredict pain. There were no effects of exposure on perceived harm. Exposure may profitably be conceived of as the learning of exceptions to a general rule. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Quality of the lipid fraction of Italian biscuits

Although not perceived as fatty foods by consumers, biscuits contain a consistent quantity of lipids. Since they are daily present in the diet, an experimental study was carried out with the aim of evaluating the quality of the lipid fraction of biscuits. Amounts of fat ranging from 75 to 231 g kg^?1, mainly constituted by refined vegetable oils and margarine, were present. A high variability in fatty acid composition was found and in some cases high amounts of trans isomers of unsaturated fatty acid were observed. In addition, HPSEC analysis of polar compounds indicated an extensive hydrolytic degradation of triacylglycerols and also a high level of oxidative degradation. The latter, demonstrated by means of the contemporary measure of oxidised triacylglycerols and of triacylglycerol oligopolymers, was comparable to that found in hydrogenated oils and fats. Copyright © 2005 Society of Chemical Industry     

Xanthan gum produced by Xanthomonas campestris from cheese whey: production optimisation and rheological characterisation

BACKGROUND: This aim of this study was the production and rheological characterisation of xanthan gum by Xanthomonas campestris pv. mangiferaeindicae IBSBF 1230 using industrial media and experimental design techniques in a bench bioreactor. RESULTS: The optimised conditions for the production of xanthan starting with 900 mL of cheese whey were 1 g L^?1 magnesium sulphate, 20 g L^?1 potassium phosphate, 28 °C temperature and initial pH 7.2 at 390 rpm agitation and 1.5 vvm aeration, resulting in 36 g L^?1 gum in 72 h. The highest viscosity obtained in the production optimisation study was 1831.34 mPa s at 25 °C with 30 g L^?1 gum. The use of CaCl₂ resulted in the highest solution viscosity under conditions of 25 °C, 1 g L^?1 salt and 46.8 g L^?1 gum, with a value of 1704.53 mPa s. CONCLUSION: In this study, cheese whey, a by‐product of the dairy industry, was used as substrate in the production of xanthan gum, a valuable product in food applications, with optimised high gum production in a bioreactor and a wide range of viscosity values. Copyright © 2009 Society of Chemical Industry     

Effects of resveratrol, piceatannol, tri-acetoxystilbene, and genistein on the inflammatory response of human peripheral blood leukocytes

Inflammatory processes are involved in the etiology of diseases. We analyzed the effect of resveratrol, piceatannol, synthetic tri-acetoxystilbene (TAS), and genistein (Bonistein(TM)) on the production of inflammatory mediators including prostaglandin E2 (PGE2), tumor necrosis factor-alpha (TNF-alpha), interleukins, and chemokines, which participate in the progression of inflammation. In order to induce inflammatory responses, human peripheral blood mononuclear and/or polymorphonuclear leukocytes were stimulated with lipopolysaccharide (LPS) and interferon-gamma (IFN(gamma)) and the production of PGE2, interleukin-8 (IL-8), and TNF-alpha was determined. In response to the stimuli, genes were substantially activated within < 2 h (e. g., TNF-alpha, IL-1alpha), or at a later stage, (e. g., COX-2, IL-6, IL-8). Unlike genistein, resveratrol and related compounds dose-dependently reduced PGE2 production. Genistein, piceatannol, and TAS diminished secretion of TNF-alpha, and IL-8. TAS reduced mRNA levels of COX-2, TNF-alpha, IL-8, IL-6, and IL-1alpha, while resveratrol impaired early expression of IL-8 and TNF-alpha. Piceatannol out-performed resveratrol, yet without matching TAS. Genistein downregulated TNF-alpha and IL-8 expression. These substances altered the LPS/IFNgamma-induced gene expression in mononuclear cells rather than in polymorphonuclear leukocytes. Immunoblot analyses corroborated the distinct activity pattern of resveratrol and genistein. In conclusion, resveratrol and their derivatives attenuated the inflammatory response of PBLs at several levels, whereas genistein acts on cytokines and pro-inflammatory interleukins.     

Pro-oxidant activity of oxidized triacylglycerols in olive oil and comparison with pro-oxidant action of polar triacylglycerol oligopolymers

An experimental investigation was carried out with the aim to evaluate the pro-oxidant activity of oxidized triacylglycerols and compare it with that observed for polar triacylglycerol oligopolymers. The research on these classes of substances, that form during processing and storage of fats and oils, has a great concern with the study of the effect of heat in technologic processes, such as vegetable oils refining, frying and cooking. Oxidized triacylglycerols and polar triacylglycerol oligopolymers, obtained in purity by preparative gel permeation chromatography, were added in amounts of 0.25-1 g/100 g both to a purified and an unpurified olive oil. Regarding the oxidized triacylglycerols, the accelerated oxidation tests showed that the induction times of both oils significantly decreased at increasing concentrations of these compounds, demonstrating their pro-oxidant effect. Moreover, the data showed that the pro-oxidant activity of oxidized triacylglycerols was greater than that of polar triacylglycerol oligopolymers.     

Wine extracts from Sardinian grape varieties attenuate membrane oxidative damage in Caco-2 cell monolayers

One of the most important sites of polyphenol action seems to be in the gastrointestinal system before absorption. We investigated the ability of three wine phenolic extracts, obtained from grape varieties grown in Sardinia, Cannonau (red), Vermentino and Malvasia (white), to exert an antioxidant action against tert-butyl hydroperoxide (TBH)-induced oxidative damage to Caco-2 cell monolayers as a model system of the human intestine. TBH treatment caused the disruption of epithelial integrity, measured as transepithelial electrical resistance, and markers of the peroxidation process of membrane lipids, MDA, fatty acid hydroperoxides and 7-ketocholesterol. All wine extracts were able to counteract the oxidising action of TBH and, in spite of the differences in phenolic composition, exerted a comparable activity. Our findings point out a direct antioxidant action of the wine extracts on enterocytes exposed to oxidising species and further support the opinion that total phenolic content is not essential for antioxidant activity.