Recognition of glycoconjugates by Helicobacter pylori. Comparison of two sialic acid-dependent specificities based on haemagglutination and binding to human erythrocyte glycoconjugates

Helicobacter pylori expresses separate binding characteristics depending on growth conditions, as documented by binding to human erythrocyte glycoconjugates. Cells grown in Ham's F12 liquid medium exhibited a selective sialic acid-dependent binding to polyglycosylceramides, PGCs (Miller-Podraza et al. (1996) Glycoconjugate J 13:453–60). There was no binding to traditional sialylated glycoconjugates like shorter-chain gangliosides, glycophorin or fetuin. However, cells grown on Brucella agar bound both to PGCs and other sialylated glycoconjugates. Fetuin was an effective inhibitor of haemagglutination caused by agar-grown cells, but had no or a very weak inhibitory effect on haemagglutination by F12-grown bacteria. PGCs were strong inhibitors in both cases, while asialofetuin was completely ineffective. The results indicate that H. pylori is able to express two separate sialic acid-dependent specificities, one represented by binding to fetuin, as described before, and another represented by a selective binding to PGCs. Abbreviations: PGCs, polyglycosylceramides; TLC, thin-layer chromatography; SDS PAGE, sodium dodecylsulfate polyacrylamide gel electrophoresis; BSA, bovine serum albumin; C, chloroform; M, methanol. The carbohydrate and glycosphingolipid nomenclatures are according to recommendations of IUPAC-IUB Commission on Biochemical Nomenclature (Lipids (1977) 12:455–68; J Biol Chem (1982) 257:3347–51 and J Biol Chem (1987) 262:13–18). This revised version was published online in November 2006 with corrections to the Cover Date.     

Different Profile of mRNA Expression in Sinoatrial Node from Streptozotocin-Induced Diabetic Rat

BackgroundExperiments in isolated perfused heart have shown that heart rate is lower and sinoatrial node (SAN) action potential duration is longer in streptozotocin (STZ)–induced diabetic rat compared to controls. In sino-atrial preparations the pacemaker cycle length and sino-atrial conduction time are prolonged in STZ heart. To further clarify the molecular basis of electrical disturbances in the diabetic heart the profile of mRNA encoding a wide variety of proteins associated with the generation and transmission of electrical activity has been evaluated in the SAN of STZ-induced diabetic rat heart.Conclusions/SignificanceCollectively, this study has demonstrated differences in the profile of mRNA encoding a variety of proteins that are associated with the generation, conduction and regulation of electrical signals in the SAN of STZ-induced diabetic rat heart. Data from this study will provide a basis for a substantial range of future studies to investigate whether these changes in mRNA translate into changes in electrophysiological function.     

Tanshinone production in roots of micropropagated Salvia przewalskii Maxim

Production of tanshinones (tanshinone I and IIA) was determined in roots of Salvia przewalskii micropropagated plants. It was found that the total tanshinone content (tashinone I and tashinone IIA) was dependent on the age of the analyzed plants. The roots of 2-year-old in vitro regenerated plants at flowering stage produced highest tanshinone levels (3.8 mg/g dry weight of tanshinone I and 7.6 mg/g dry weight of tanshinone IIA).     

Lectin chromatography/mass spectrometry discovery workflow identifies putative biomarkers of aggressive breast cancers

We used a lectin chromatography/MS-based approach to screen conditioned medium from a panel of luminal (less aggressive) and triple negative (more aggressive) breast cancer cell lines (n=5/subtype). The samples were fractionated using the lectins Aleuria aurantia (AAL) and Sambucus nigra agglutinin (SNA), which recognize fucose and sialic acid, respectively. The bound fractions were enzymatically N-deglycosylated and analyzed by LC-MS/MS. In total, we identified 533 glycoproteins, ～90% of which were components of the cell surface or extracellular matrix. We observed 1011 glycosites, 100 of which were solely detected in ≥3 triple negative lines. Statistical analyses suggested that a number of these glycosites were triple negative-specific and thus potential biomarkers for this tumor subtype. An analysis of RNaseq data revealed that approximately half of the mRNAs encoding the protein scaffolds that carried potential biomarker glycosites were up-regulated in triple negative vs luminal cell lines, and that a number of genes encoding fucosyl- or sialyltransferases were differentially expressed between the two subtypes, suggesting that alterations in glycosylation may also drive candidate identification. Notably, the glycoproteins from which these putative biomarker candidates were derived are involved in cancer-related processes. Thus, they may represent novel therapeutic targets for this aggressive tumor subtype.     

Genetic variability of body weight in two goose strains under long-term selection

Body weight is one of the most important traits in any genetic improvement program in geese for at least 2 reasons. First, measurements of the trait are very easy. Second, body weight is correlated with a number of other meat performance traits. However, the genetic background of body weight shows considerable complexity. Three genetic models (with direct, maternal genetic and permanent maternal environmental effects) were employed in this study. Records of 3076 individuals of maternal strain W11 and 2656 individuals of paternal strain W33 over 6 consecutive generations, kept in the pedigree farm of Ko?uda Wielka, were analysed. Body weight (in kilograms) was measured in weeks 8 (BW8) and 11 (BW11). The inbreeding levels in both populations were relatively low (0.14% and 0.02% for W11 and W33, respectively), therefore these effects were not included in the linear models to estimate genetic parameters. Three fixed effects (hatch period, sex and year) were included in each linear model. Two criteria (AIC, BIC) were used to check the goodness of fit of the models. The computations were performed by WOMBAT software. In general, the genetic parameter estimates varied across the traits, models and strains studied. Direct additive heritability estimates ranged from 0.0001 (for BW11 of W33) to 0.55 (for BW11 of W33). Maternal and total heritabilities were also variable. Estimates of ratios of direct-maternal effect covariance in phenotypic variance were both positive and negative, but they were negligible, whereas ratios of the permanent environmental maternal variance to phenotypic variance were close to zero. Both of the applied criteria of model adequacy indicate that the model with maternal genetic and environmental effects should be considered as optimal. Genetic trends were close to zero. It seems that they were influenced by long-term selection. Similar tendencies have been observed for phenotypic trends, as well.     

The Influence of the Occupational Exposure to Heavy Metals and Tobacco Smoke on the Selected Oxidative Stress Markers in Smelters

The aim of the study was to verify if there is any association between exposure to Cu, Zn, Cd, Pb, As and the formation of malondialdehyde (MDA), 8-hydroxydeoxyguanosine (8-OHdG), advanced oxidation protein products (AOPP), and whether in this process cigarette smoking plays a role. The investigations were performed in the 352 smelters occupationally exposed to heavy metals and 73 persons of control group. Metals concentration was determined by atomic absorption spectrometry. MDA and AOPP concentrations were determined by spectrophotometric methods. The concentration of 8-OHdG was determined by ELISA method. It was demonstrated an increased Cu concentration in smoking smelters compared to non-smoking control group. It was noted no differences in Zn and Mg concentrations between the examined groups. Pb concentration was more than sixfold higher in the group of smoking smelters and about fivefold higher in the group of non-smoking smelters compared to the control groups (smokers and non-smokers). It was shown that Cd concentration in the blood was nearly fivefold higher in the smoking control group compared to the non-smoking control group and more than threefold higher in the group of smoking smelters compared to non-smoking. It was shown an increased As concentration (more than fourfold) and decreased Ca concentration in both groups of smelters compared to control groups. In groups of smelters (smokers and non-smokers), twofold higher MDA and AOPP concentrations, and AOPP/albumin index compared to control groups (smokers and non-smokers) were shown. Tobacco smoke is the major source of Cd in the blood of smelters. Occupational exposure causes accumulation of Pb in the blood. Occupational exposure to heavy metals causes raise of MDA concentration and causes greater increase in AOPP concentration than tobacco smoke.     

Lymphocyte responses of rats vaccinated with cDNA encoding a phosphoglycerate kinase of Fasciola hepatica (FhPGK) and F. hepatica infection

Lymphocyte responses in the blood, peritoneal fluid and both mesenteric and hepatic lymph nodes of cDNA-FhPGK/pCMV vaccinated and/or Fasciola hepatica infected rats of both sexes were investigated to provide an insight into the immune responses that develop in different body compartments. The immune response that developed in cDNA-FhPGK/pCMV vaccinated females contributed to partial protection against F. hepatica infection (54% reduction in fluke recovery), while more liver flukes were found in the livers and bile ducts of cDNA-FhPGK/pCMV vaccinated male rats than in unvaccinated animals (increase of 13%). Rat sex not only affected the ultimate effectiveness of vaccination but also lymphocyte responses following vaccination and/or infection. Different CD4 + and CD8 + T cell profiles were noted in peritoneal fluid and lymph nodes, but not in blood, during acute and chronic fasciolosis. Moreover, independent lymphocyte responses developed in distinct body compartments. Immune responses of rats were polarized towards Th2/Treg with lymphocytes isolated from male rats showing higher IL-4 and IL-10 production than females. Lymphocyte proliferative capacities in response to mitogen (PHA) or vaccine antigen (FhPGK) were impaired in both sexes with a considerably higher reduction observed for males and restored lymphocyte proliferative capacities reported for females vaccinated with cDNA-FhPGK/pCMV during chronic fasciolosis.     

Effect of chronic vanadium administration in drinking water to rats

Two-month old Wistar rats of both sexes received, as sole drinking liquid, an aqueous solution of ammonium metavanadate (AMV) at a concentration of 0.01 or 0.05 mg V cm^–3 (0.2 or 1.0 mM) for a period of 4 weeks. It was calculated that the animals took up doses of 1.5 and 5–6 mg V kg body weight^–1 24 h^–1, respectively. Food and AMV solution consumption in the experimental groups was similar to food and water consumption in the control group. A statistically significant decrease of consumption of AMV solution at a concentration of 0.05 mg V cm^–3 was noted only in males. Hematological examination demonstrated a decrease in the erythrocyte count, hemoglobin level and hematocrit index. This decrease in the erythrocyte count was associated with an increased percentage of reticulocytes in the peripheral blood of the animals drinking the solution with a higher vanadium content. Biochemical analyses demonstrated a decrease of l-ascorbic acid levels in the plasma and erythrocytes of animals drinking the AMV solutions. A distinct tendency for the malonyldialdehyde level to increase in the blood was also observed. Among the enzymes examined in the erythrocytes (catalase, glucose-6-phosphate dehydrogenase, lactate dehydrogenase and -aminolevulinic acid dehydratase [ALA-D]) only ALA-D activity was depressed.