Characterization of the CD14⁺⁺CD16⁺ Monocyte Population in Human Bone Marrow

Numerous studies have divided blood monocytes according to their expression of the surface markers CD14 and CD16 into following subsets: classical CD14(++)CD16(-), intermediate CD14(++)CD16(+) and nonclassical CD14(+)CD16(++) monocytes. These subsets differ in phenotype and function and are further correlated to cardiovascular disease, inflammation and cancer. However, the CD14/CD16 nature of resident monocytes in human bone marrow remains largely unknown. In the present study, we identified a major population of CD14(++)CD16(+) monocytes by using cryopreserved bone marrow mononuclear cells from healthy donors. These cells express essential monocyte-related antigens and chemokine receptors such as CD11a, CD18, CD44, HLA-DR, Ccr2, Ccr5, Cx3cr1, Cxcr2 and Cxcr4. Notably, the expression of Ccr2 was inducible during culture. Furthermore, sorted CD14(++)CD16(+) bone marrow cells show typical macrophage morphology, phagocytic activity, angiogenic features and generation of intracellular oxygen species. Side-by-side comparison of the chemokine receptor profile with unpaired blood samples also demonstrated that these rather premature medullar monocytes mainly match the phenotype of intermediate and partially of (non) classical monocytes. Together, human monocytes obviously acquire their definitive CD14/CD16 signature in the bloodstream and the medullar monocytes probably transform into CD14(++)CD16(-) and CD14(+)CD16(++) subsets which appear enriched in the periphery

Role of heparan sulfate for attachment and entry of tick-borne encephalitis virus

AbstractAttachment of the flavivirus tick-borne encephalitis (TBE) virus to different permissive cell lines was investigated by a newly established quantitative assay using fluorescence-labeled virus. Previous work had shown that BHK-21 cell-adapted mutants of TBE virus had acquired potential heparan sulfate (HS) binding sites on the outer surface of protein E. Quantitative analysis of one of these mutants indicated that it attached to HS-expressing cell lines with a 10- to 13-fold higher affinity than wild-type TBE virus strain Neudoerfl. CHO cells deficient in HS synthesis bound less than 5% of the amount of wild-type or mutant virus that could attach to HS-containing CHO cells but were nevertheless found to be highly susceptible to infection with both viruses. Thus, even though HS is a major determinant of TBE virus attachment on HS-expressing cells, our findings suggest the existence of an alternative host cell receptor that is less abundant than HS

Determination of Glutathione in Austrian Wine Samples: The Effects of Freezing, the Choice of Yeast and Storage

Glutathione (GSH, γ-L-Glutamyl-L-Cysteinyl Glycine) is a tripeptide of L-glutamate, L-cysteine and glycine. GSH in wine is derived from either grapes or yeast, during alcoholic fermentation. The GSH concentration in wine is very variable and depends on the environmental conditions as well as viticultural practices. During winemaking GSH has a significant role in oxidation prevention due to its unique redox and nucleophilic properties. Since GSH is very reactive it is highly important to prepare samples immediately and under inert conditions just prior to the determination of the GSH concentration. Therefore the aim of this research was to implement a method for the quantitative determination of GSH levels in grape juices (musts) made from different Austrian grape varieties and to investigate the influence of yeast on the GSH content in wine after aging. The results of this research have shown that monitoring with nitrogen gas, sulphur dioxide and freezing process at −25 ºC led to a good protective effect on the free glutathione amount in wine and grape samples. The GSH concentration in the samples was variable. Levels were ranging from non-detectable to up to 23.10 mg/l, and it showed that grape variety has no impact on GSH concentration in the must. Furthermore the results suggest that the choice of yeast has an impact on GSH content in wine even after 6 and 18 months of aging

Fundamental Valuation of Extra-Financial Information

We augment seminal models based on Ohlson (1995) by integrating the value impact of ratings related to three different extra-financial categories, i. e. corporate governance, human capital, and innovation capital. By integrating extra-financial information in valuation models, we examine whether current market values can be better estimated and future stock performance better predicted when considering this information. For a sample of large Euro-pean public firms, we find that a model including human capital information and analysts’ earnings forecasts best explains current stock prices. Our model based on human capital in-formation (without analysts’ forecasts) best identifies under- and overvalued companies

Functional microRNA generated from a cytoplasmic RNA virus

MicroRNAs (miRNAs) are a class of small, non-coding RNAs that play a pivotal role in the regulation of posttranscriptional gene expression in a wide range of eukaryotic organisms. Although DNA viruses have been shown to encode miRNAs and exploit the cellular RNA silencing machinery as a convenient way to regulate viral and host gene expression, it is generally believed that this pathway is not available to RNA viruses that replicate in the cytoplasm of the cell because miRNA biogenesis is initiated in the nucleus. In fact, among the >200 viral miRNAs that have been experimentally verified so far, none is derived from an RNA virus. Here, we show that a cytoplasmic RNA virus can indeed encode and produce a functional miRNA. We introduced a heterologous miRNA-precursor stem-loop sequence element into the RNA genome of the flavivirus tick-borne encephalitis virus, and this led to the production of a functional miRNA during viral infection without impairing viral RNA replication. These findings demonstrate that miRNA biogenesis can be used by cytoplasmic RNA viruses to produce regulatory molecules for the modulation of the transcriptome

Impact of school lunch type on nutritional quality of English children's diets.

OBJECTIVE: Nutrient and food standards exist for school lunches in English primary schools although packed lunches brought from home are not regulated. The aim of the present study was to determine nutritional and dietary differences by lunch type. DESIGN: A cross-sectional survey was carried out in 2007 assessing diet using the Child and Diet Evaluation Tool (CADET), a validated 24 h estimated food diary. The data were analysed to determine nutritional and dietary intakes over the whole day by school meal type: school meals and packed lunches. SETTING: Fifty-four primary schools across England. SUBJECTS: Children (n 2709) aged 6-8 years. RESULTS: Children having a packed lunch consumed on average 11·0 g more total sugars (95 % CI 6·6, 15·3 g) and 101 mg more Na (95 % CI 29, 173 mg) over the whole day. Conversely, children having a school meal consumed, on average, 4·0 g more protein (95 % CI 2·3, 5·7 g), 0·9 g more fibre (NSP; 95 % CI 0·5, 1·3 g) and 0·4 mg more Zn (95 % CI 0·1, 0·6 mg). There was no difference in daily energy intake by lunch type. Children having a packed lunch were more likely to consume snacks and sweetened drinks; while children having a school meal were more likely to consume different types of vegetables and drink water over the whole day. CONCLUSIONS: Compared with children having a school meal, children taking a packed lunch to school consumed a lower-quality diet over the whole day, including higher levels of sugar and Na and fewer vegetables. These findings support the introduction of policies that increase school meal uptake

Mutations at positions 186 and 194 in the HA gene of the 2009 H1N1 pandemic influenza virus improve replication in cell culture and eggs

Obtaining suitable seed viruses for influenza vaccines poses a challenge for public health authorities and manufacturers. We used reverse genetics to generate vaccine seed-compatible viruses from the 2009 pandemic swine-origin influenza virus. Comparison of viruses recovered with variations in residues 186 and 194 (based on the H3 numbering system) of the viral hemagglutinin showed that these viruses differed with respect to their ability to grow in eggs and cultured cells. Thus, we have demonstrated that molecular cloning of members of a quasispecies can help in selection of seed viruses for vaccine manufacture

Vascular CXCR4 Limits Atherosclerosis by Maintaining Arterial Integrity Evidence From Mouse and Human Studies

BACKGROUND: The CXCL12/CXCR4 chemokine ligand/receptor axis controls (progenitor) cell homeostasis and trafficking. So far, an atheroprotective role of CXCL12/CXCR4 has only been implied through pharmacological intervention, in particular, because the somatic deletion of the CXCR4 gene in mice is embryonically lethal. Moreover, cell-specific effects of CXCR4 in the arterial wall and underlying mechanisms remain elusive, prompting us to investigate the relevance of CXCR4 in vascular cell types for atheroprotection. METHODS: We examined the role of vascular CXCR4 in atherosclerosis and plaque composition by inducing an endothelial cell (BmxCreERT2-driven)-specific or smooth muscle cell (SMC, SmmhcCreERT2-or TaglnCre-driven)-specific deficiency of CXCR4 in an apolipoprotein E-deficient mouse model. To identify underlying mechanisms for effects of CXCR4, we studied endothelial permeability, intravital leukocyte adhesion, involvement of the Akt/WNT/beta-catenin signaling pathway and relevant phosphatases in VE-cadherin expression and function, vascular tone in aortic rings, cholesterol efflux from macrophages, and expression of SMC phenotypic markers. Finally, we analyzed associations of common genetic variants at the CXCR4 locus with the risk for coronary heart disease, along with CXCR4 transcript expression in human atherosclerotic plaques. RESULTS: The cell-specific deletion of CXCR4 in arterial endothelial cells (n=1215) or SMCs (n=13-24) markedly increased atherosclerotic lesion formation in hyperlipidemic mice. Endothelial barrier function was promoted by CXCL12/\CXCR4, which triggered Akt/WNT/beta-catenin signaling to drive VE-cadherin expression and stabilized junctional VE-cadherin complexes through associated phosphatases. Conversely, endothelial CXCR4 deficiency caused arterial leakage and inflammatory leukocyte recruitment during atherogenesis. In arterial SMCs, CXCR4 sustained normal vascular reactivity and contractile responses, whereas CXCR4 deficiency favored a synthetic phenotype, the occurrence of macrophage-like SMCs in the lesions, and impaired cholesterol efflux. Regression analyses in humans (n=259 796) identified the C-allele at rs2322864 within the CXCR4 locus to be associated with increased risk for coronary heart disease. In line, C/C risk genotype carriers showed reduced CXCR4 expression in carotid artery plaques (n=188), which was furthermore associated with symptomatic disease. CONCLUSIONS: Our data clearly establish that vascular CXCR4 limits atherosclerosis by maintaining arterial integrity, preserving endothelial barrier function, and a normal contractile SMC phenotype. Enhancing these beneficial functions of arterial CXCR4 by selective modulators might open novel therapeutic options in atherosclerosis