Cow's milk allergy (CMA) has become a common disease in early childhood, its prevalence ranging from 1.6% to 2.8% among children younger than 2 years of age. The role of different cow's milk protein (CMP) in the pathogenesis of CMA is still controversial. Even if the proteins most frequently and most intensively recognized by immunoglobulin E (IgE) seem to be the most abundant in milk (caseins and beta-lactoglobulin), with an although great variability all milk proteins appear to be potential allergens, even those that are present in trace amounts (i.e., lactoferrin, IgG, and BSA). In this work proteomics techniques have been applied for CMP allergens analysis. Allergens have been identified by immunoblotting following resolution of CMP components by two-dimensional electrophoresis. Sera from 20 milk-allergic subjects, as proven by oral provocation test, CAP-RAST and skin prick test, have been used for cow's milk major allergen identification. Cow's milk proteins and their isoforms were identified by matrix assisted laser desorption/ionization-time of flight (MALDI-TOF)-mass spectrometry. In our group of patients, the prevalence of CMP allergens, i.e., the total number of subjects sensitized to CMP divided by the total number of the subjects enrolled in the study, was: 55% alpha(s1)-casein, 90% alpha(s2)-casein, 15% beta-casein, 50% kappa-casein, 45% beta-lactoglobulin, 45% BSA, 95% IgG-heavy chain, 50% lactoferrin, and 0% alpha-lactalbumin.

Aims:  The goal of this work was to investigate the influence of DMSO, garlic extract and p‐coumaric acid on bacterial quorum sensing (QS). Methods and Results:  The decreases in the QS responses of QS reporter strains Escherichia coli pSB401 and pSB536, Agrobacterium tumefaciens NTL4, Chromobacterium violaceum 5999 and wt 494, Pseudomonas putida IsoF/gfp and environmental Pseudomonas chlororaphis were quantified in relation to growth inhibitory effects. DMSO showed no significant QS‐specific effects on the strains tested even at close‐to‐lethal concentrations. Garlic extracts antagonized the activity of QS receptors LuxR, AhyR and TraR, but were toxic at higher concentrations. P‐coumaric acid fully inhibited QS responses of 5999, NTL4 and P. chlororaphis, with no influence on cell viability. Conclusions:  The quorum sensing inhibition activity of garlic was extended to novel receptors, and p‐coumaric acid was found to possess previously undescribed QS antagonist properties. Significance and Impact of the Study:  The results suggest that p‐coumaric acid might act as QS inhibitor. Further studies are required to understand its role in the regulation of QS and investigate structurally related compounds.

This study analyzes the effect of chronic treatment with different antioxidants (N-acetyl-cysteine [NAC], taurine, a combination of NAC and taurine, and oxerutin) on long-term experimental diabetes induced by streptozotocin in rats. Glycoxidative damage was evaluated in the skin; glomerular structural changes were studied with morphometry and immunohistochemistry. Oxerutin treatment and the combined NAC plus taurine treatment resulted in reduced accumulation of collagenlinked fluorescence in skin in comparison with untreated diabetic rats. All treatments except taurine reduced glomerular accumulation of N ⑀ -(carboxymethyl)lysine and protected against the increase in glomerular volume typical of diabetes; furthermore, the apoptosis rate was significantly decreased and the glomerular cell density was better preserved. Glycoxidative markers in the skin turned out to be good indicators of the glomerular condition. The findings that emerged from our study support the hypothesis that glomerular damage in diabetes can be prevented or at least attenuated by supplementation with specific antioxidants. Treatment with oxerutin and combined treatment with NAC plus taurine gave the most encouraging results, whereas the results of taurine-only treatment were either negligible or negative and therefore suggest caution in the use of this molecule in single-drug treatment courses. Diabetes 52: 499 -505, 2003

Background The main functional property of collagen is to provide a supporting framework to almost all tissues: the effects of non‐enzymatic glycation on this protein are deleterious and in diabetes mellitus contribute to the mechanism of late complications. The aim of this work is to provide evidence by scanning force microscopy of modifications in collagen structure caused by high glucose concentration, in vivo and in vitro, and to correlate the data with markers of non‐enzymatic glycation. Methods Tendon fibrils were obtained from the tails of 8‐month‐old rats (BB/WOR/MOL\BB) which developed diabetes spontaneously at least 12 weeks before they were killed, and from diabetes‐resistant rats of the same strain (BB/WOR/MOL\WB). A scanning force microscope (SFM; Nanoscope III) equipped with a Contact Mode Head was used for imaging. Band interval, diameter and depth of D‐band gap were measured in non‐diabetic and diabetic tail tendon fibrils and in fibrils incubated with glucose (0.5 M for 2 weeks). Fructosamine was determined in the tendon fibrils by a colorimetric method and pentosidine was evaluated in acid‐hydrolyzed samples by coupled reverse phase‐ionic exchange column HPLC. Results Incubated fibrils revealed modifications in radius (228±5 nm) and gap depth (3.65±0.10 nm) that closely reproduce diabetes‐induced damage (236±3 and 3.20±0.04 nm respectively) and were significantly different from the pattern seen in non‐diabetic fibrils (151±1 and 2.06±0.03 nm; p<0.001). Both fructosamine and pentosidine were higher in diabetic (3.82±1.43 nmol/mg and 2.23±0.24 pmol/mg collagen respectively) and in glucose‐incubated fibrils (9.27±0.55 nmol/mg and 5.15±0.12 pmol/mg collagen respectively) vs non‐diabetic tendons (1.29±0.08 nmol/mg and 0.88±0.11 pmol/mg collagen respectively; p<0.01); during the time course of incubation, an early increase in fructosamine was seen, whereas pentosidine increased later. The D‐band parameter was similar in all three groups, indicating that axial organization is not modified by non‐enzymatic glycation. Conclusion This is the first description obtained with SFM of diabetes‐induced ultrastructural changes in collagen fibrils. Moreover, the data presented are consistent with the concept that chronic exposure of collagen to glucose in vivo or in vitro leads to similar structural modifications in collagen fibrils, probably through crosslinks. The correlation between morphologic parameters and both markers of glycation provides strong evidence for a crucial role of this non‐enzymatic modification. Copyright © 2000 John Wiley & Sons, Ltd.

Abstract. Progressive supranuclear palsy (PSP) is a neurodegenerative disorder characterized by extensive neurofibrillary tangle (NFT) formation and neuronal loss in selective neuronal populations. Currently, no clues to the biological events underlying the pathological process have emerged. In Alzheimer disease (AD), which shares with PSP the occurrence of NFTs, advanced glycation end products (AGEs) as well as oxidation adducts have been found to be increased in association with neurofibrillary pathology. The presence and the amount of lipid and protein oxidation markers, as well as of pyrraline and pentosidine, 2 major AGEs, was assessed by biochemical, immunochemical, and immunocytochemical analysis in midbrain tissue from 5 PSP cases, 6 sporadic AD cases, and 6 age-matched control cases. The levels of 4-hydroxynonenal (HNE) and thiobarbituric acid reactive substances (TBARS), 2 major products of lipid peroxidation, were significantly increased by 1.6-fold (p Ͻ 0.04) and 3.9-fold (p Ͻ 0.01), respectively, in PSP compared with control tissues, whereas in AD only TBARS were significantly increased. In PSP tissue the intensity of neuronal HNE immunoreactivity was proportional to the extent of abnormal aggregated protein. The amount of protein oxidation products and AGEs was instead similar in PSP and control tissues. In AD, a higher but not significant level of pyrraline and pentosidine was measured, whereas the level of carbonyl groups was doubled. These findings indicate that in PSP, unlike in AD, lipid peroxidation is selectively associated with NFT formation. The intraneuronal accumulation of toxic aldehydes may contribute to hamper degradation, leading to its aggregation in the PSP specific abnormal filaments.