Anti-Helicobacter pylori effects of IgY from egg york of immunized hens

Effects of egg york containing IgY specific for Helicobacter pylori on the bacterial growth and intragastric infection were investigated in comparison with a proton-pump inhibitor pantoprazole. For in vitro anti-bacterial activity test, H. pylori (1×108 CFU/mL) was incubated with a serially diluted IgY for 3 days. As a result, IgY fully inhibited the bacterial growth at 16 mg/mL, which was determined to a minimal inhibitory concentration. In vivo elimination study, male C57BL/6 mice were infected with the bacteria by intragastric inoculation (1×108 CFU/mouse) 3 times at 2-day intervals, and 2 weeks later, orally treated twice a day with 50, 100, 200 or 500 mg/kg IgY for 18 days. After the final administration, biopsy sample of the gastric mucosa was assayed for the bacterial identification via urease, oxidase, catalase, nitrate reduction and H2S tests in addition to microscopic examination for mucosal inflammation. In CLO kit test, 75, 50, 12.5 and 12.5% of the animals revealed positive reaction following treatment with 50, 100, 200 and 500 mg/kg IgY, respectively, resulting in a superior efficacy at 200 mg/kg than 30 mg/kg pantoprazole that displayed 75% elimination. The CLO test results were confirmed by bacterial identification. Microscopic examination revealed that H. pylori infection caused severe gastric mucosal inflammation, which were not observed in the CLO-negative mice following treatment with IgY or pantoprazole. Taken together, IgY inhibited the growth of H. pylori, and improved gastritis and villi injuries by eliminating the bacteria from the stomach. The results indicate that IgY could be a good candidate overcoming tolerance of antibiotics for the treatment of H. pylori-mediated gastric ulcers

Vergleich der Rezidivraten bei Basalzellkarzinomen im Kopf-Hals-Bereich R0 und R1 Resektion

Das Basalzellkarzinom ist heute einer der häufigsten Tumore des Menschen und der häufigste in der westlichen Welt. Die Relevanz ist für diesen Tumor mit Millionen von neuen Fällen weltweit jährlich und einer ansteigenden Inzidenz sowie Todesfällen sehr hoch. Der wichtigste Risikofaktor dieses Tumors, der ohne Vorstufe besonders an lichtexponierten Körperstellen und zu 80 % im Kopf- und Halsbereich auftritt, ist die ultraviolette Bestrahlung. Bei diesem Tumor gibt es verschiedene histologische Typen, die unterschiedliche Relevanz bezüglich Tumorbehandlung und Tumorrezidiv besitzen. Am häufigsten tritt der noduläre Typ auf. Die Rezidive sind relativ selten, aber dann von großer Relevanz, da es meist aggressivere und schwieriger zu behandelnde Tumoren sind. Behandlungsmethode der ersten Wahl ist die chirurgische, wobei die mikrographisch kontrollierte Chirurgie eine sensible, gewebesparende Methode ist und eine lückenlose Schnittrandbeurteilung des Tumorexzidates im Vergleich mit der konventionellen Chirurgie bietet. Die Rezidivraten unterscheiden sich aufgrund verschiedener Faktoren wesentlich. Ziel der vorliegenden Arbeit ist die Untersuchung dieser Parameter und die Abklärung ihrer Rolle bezüglich eines Rezidivs. Die wichtigsten Parameter, deren Rolle in Bezug auf Rezidive und Rezidivraten in dieser Arbeit untersucht und verglichen werden, sind positive Schnittränder mit Residualtumoren (R1) im Vergleich mit von Residualtumoren freien Schnitträndern (R0)

Mucin Degradation Mechanisms by Distinct Pseudomonas aeruginosa Isolates In Vitro

Pseudomonas aeruginosa has emerged as an important causative agent of bacterial keratitis, a rapidly progressive ocular condition that may result in blindness. Secretory mucin forms the main constituent of the precorneal tear film, a three-layer film on the ocular surface protecting the underlying corneal epithelium from potential pathogens. The purpose of the present study was to compare mucin degradation mechanisms between ocular P. aeruginosa strains. Mucin degradation was assessed by agarose electrophoresis, lectin blotting, and size exclusion chromatography. The results indicate that certain P. aeruginosa strains (Paer12, ATCC 15442, 6294, and Paer25) had depleted mucin from the culture supernatant and that this was contingent on the inherent ability of these isolates to produce proteases. Non-protease-producing strains (Paer1 and Paer3) did not appreciably degrade mucin. Further, galactosidase, N-acetylglucosaminidase, and N-acetylgalactosaminidase activities were detected in some strains, suggesting the operation of further mechanisms of mucin degradation by P. aeruginosa. Mucin degradation by P. aeruginosa also seemed to be for the acquisition of nutrients, as a growth advantage was observed in mucin-depleting strains over nondepleting strains in the long term. It is postulated that the degradation of mucin serves to collapse the mucin barrier and its associated network containing antibacterial tear components and to provide energy for sustained bacterial growth

HPLC analysis of discrete haptoglobin isoform N-linked oligosaccharides following 2D-PAGE isolation

Glycosylation is a common but variable modification that regulates glycoprotein structure and function. We combined small format 2D-PAGE with HPLC to analyse discrete human haptoglobin isoform N-glycans. Seven major and several minor haptoglobin isoforms were detected by 2D-PAGE. N-Glycans released from Coomassie-stained gel spots using PNGase were labeled at their reducing termini with 2-aminobenzamide. HPLC analysis of selected major isoform N-glycans indicated that sialic acid composition determined their Separation by isoelectric focussing. N-Glycans from two doublets of quantitatively minor isoforms were also analysed. Although separation of each pair of doublets was influenced by sialylation, individual spots within each doublet contained identical N-glycans. Thus, heterogeneity in minor haptoglobin isoforms was due to modifications distinct from N-glycan structure. These studies describe a simple method for analysing low abundance protein N-glycans and provide details of discrete haptoglobin isoform N-glycan structures which will be useful in proteomic analysis of human plasma samples. (c) 2006 Elsevier Inc. All rights reserved

Evaluation of endogenous plasma peptide extraction methods for mass spectrometric biomarker discovery

Peptides have a role in the inflammatory response, tumor biology, and endocrine processes, presenting them as appealing biomarker candidates. However, peptide extraction efficacy for clinical profiling remains a pivotal technological challenge, as maximum coverage of the plasma peptidome is limited by a range of factors including the inherent complexity of human plasma and the lower concentration of peptides compared to abundant proteins. The aim of this study was to evaluate commonly employed peptide extraction methodologies in terms of total number of peptides detected and the mass range of peptides observed by MALDI. Despite showing coelution of proteins, solid-phase extraction (SPE) methods exhibited superior plasma peptide recovery than ultrafiltration, acetonitrile (ACN) precipitation, or size-exclusion chromatography methods under conditions employed in the study. Not surprisingly, in line with studies challenging the veracity of many peptide biomarker studies, the majority of identified peptides eluted from SPE methods corresponded to proteolytic truncations of the most abundant plasma proteins. The prefractionation of plasma with acetonitrile precipitation prior to SPE provided distinct ion signal profiles and is worthy of further study. In conclusion, this study favors the use of SPE in peptide extraction protocols for increased biomarker coverage and diversity from the plasma peptidome.11 page(s