Differences in zinc status between patients with osteoarthritis and osteoporosis

Zinc has been suggested to play an important role in the development of osteoporosis, whereas the influence of zinc on osteoarthritis has attracted much less attention. The aim of the study was to investigate and compare the zinc status and bone turnover, density, and biomechanical properties of osteoarthritic and osteoporotic patients. The study comprised 40 women who underwent hip replacement due to osteoarthritis or osteoporosis. Serum and urine zinc content, and bone resorption markers and serum bone formation markers were determined. The unaffected hip and the exarticulated affected femoral head underwent DEXA scanning. Bone biopsies were obtained from the femoral heads and the biomechanical properties were determined. The biopsies were ashed and the bone zinc content was ascertained. Osteoarthritic patients had significantly higher serum zinc concentrations and lower urine zinc concentrations than osteoporotic patients, whereas the bone zinc content did not differ. The zinc status was not found to be a predictor for the bone strength. In conclusion, the finding that the zinc status of osteoporotic patients is significantly different from that of osteoartritic patients is new and supports the view that osteoporosis and osteoarthritis rarely occur in the same individual.     

Linkage disequilibrium ofMHC genes in the chicken

Two loci of the chicken MHC (theB complex) are expressed in erythrocytes,B-F andB-G. The former is homologous to the murineK andD loci and is also expressed in all white blood cells, while the latter is apparently restricted to RBC and is of unknown relationship toH-2 andHLA loci. A recombinant between two congenic, MHC-different strains, CB and CC, has permitted the production of antisera specific for theB-F andB-G alleles of these two strains, and these and other antisera have been used for typing of outbred populations of various chicken breeds.The cross-reactions found with haplotypes other than the donors' are extensive, sometimes even extreme, but it is possible to narrow the specificity of the typing sera by appropriate absorptions. With absorbed sera we have found a linkage disequilibrium which is almost certainly even stronger than that reported in studies of mammalianMHC loci. We have also made observations which suggest that the gametic association of a given set ofB-F andB-G alleles is probably not merely a matter of random crossing-over events.     

LYST Affects Lysosome Size and Quantity,but not Trafficking or Degradation Through Autophagy or Endocytosis

Mutations in the large BEACH domain‐containing protein LYST causes Chediak–Higashi syndrome. The diagnostic hallmark is enlarged lysosomes and lysosome‐related organelles in various cell types. Dysfunctional secretion of enlarged lysosome‐related organelles has been observed in cells with mutations in LYST, but the capacity of the enlarged lysosomes to degrade endogenous proteins has not been studied. Here, we show for the first time that small interfering RNA‐depletion of LYST in human cell lines recapitulates the LYST mutant phenotype of enlarged lysosomes. We found no evidence for an effect of LYST depletion on autophagy or endocytic degradation. Autophagosomes are formed in normal size and quantities and are able to fuse to the enlarged lysosomes, leading to normal rates of degradation. Degradation of the epidermal growth factor receptor (EGFR) was similarly not affected, indicating that the enlarged lysosomes are fully functional in degrading endogenous proteins. Retrograde trafficking of toxins as well as the localization of transporters of lysosomal proteins, adaptor protein‐3 (AP‐3) and cation‐independent mannose‐6‐phosphate receptor (CI‐MPR), were all found to be unaffected by LYST. Quantitative analysis of the enlarged lysosomes shows that LYST depletion causes a reduction in vesicle quantity per cell, while the total enzymatic content and vesicular pH are unaffected, supporting a role for LYST in lysosomal fission and/or fusion events.      

Segregated Phases in Pulmonary Surfactant Membranes Do Not Show Coexistence of Lipid Populations with Differentiated Dynamic Properties

The composition of pulmonary surfactant membranes and films has evolved to support a complex lateral structure, including segregation of ordered/disordered phases maintained up to physiological temperatures. In this study, we have analyzed the temperature-dependent dynamic properties of native surfactant membranes and membranes reconstituted from two surfactant hydrophobic fractions (i.e., all the lipids plus the hydrophobic proteins SP-B and SP-C, or only the total lipid fraction). These preparations show micrometer-sized fluid ordered/disordered phase coexistence, associated with a broad endothermic transition ending close to 37°C. However, both types of membrane exhibit uniform lipid mobility when analyzed by electron paramagnetic resonance with different spin-labeled phospholipids. A similar feature is observed with pulse-field gradient NMR experiments on oriented membranes reconstituted from the two types of surfactant hydrophobic extract. These latter results suggest that lipid dynamics are similar in the coexisting fluid phases observed by fluorescence microscopy. Additionally, it is found that surfactant proteins significantly reduce the average intramolecular lipid mobility and translational diffusion of phospholipids in the membranes, and that removal of cholesterol has a profound impact on both the lateral structure and dynamics of surfactant lipid membranes. We believe that the particular lipid composition of surfactant imposes a highly dynamic framework on the membrane structure, as well as maintains a lateral organization that is poised at the edge of critical transitions occurring under physiological conditions.     

Vegetation transition following drainage in a high‐latitude hyper‐oceanic ecosystem

Questions: How does draining affect the composition of vegetation? Are certain functional groups favoured? Can soil parameters explain these differences? Location: Central Faroe Islands, treeless islands in the northern boreal vegetation zone. Since 1987, an area of 21 km² at 100–200 m a.s.l. was drained in order to provide water for hydro‐electric production. Method: Vegetation and soil of a drained area and a control, undrained neighbouring area of approximately the same size were sampled in 2007. Six sites were sampled in each area. The vegetation was classified with cluster analysis. Results: Four plant communities were defined in the area: Calluna vulgaris–Empetrum nigrum–Vaccinium myrtillus heath, Scirpus cespitosus–Eriophorum angustifolium blanket mire, Carex bigelowii–Racomitrium lanuginosum moss‐heath, Narthecium ossifragum–Carex panacea mire. Heath was more extensively distributed within, and was the dominant community of the drained area, whereas moss‐heath was more extensive in the undrained area. Blanket mire and mire had approximately the same distribution in both areas. For the blanket mire, species composition indicated drier conditions in the drained than in the undrained area. The drained area had higher frequencies of woody species and lichens, grasses had finer roots and available soil phosphate was considerably higher, whereas the undrained area had higher frequencies of grasses and sedges. Conclusion: The dominant plant communities were different in the two areas, which indicated that the blanket mire was drying in the drained area. Higher concentration of soil phosphate in the drained area also indicated increased decomposition of organic soils owing to desiccation.     

A dual function for Deep orange in programmed autophagy in the Drosophila melanogaster fat body

Lysosomal degradation of cytoplasm by way of autophagy is essential for cellular amino acid homeostasis and for tissue remodeling. In insects such as Drosophila, autophagy is developmentally upregulated in the larval fat body prior to metamorphosis. Here, autophagy is induced by the hormone ecdysone through down-regulation of the autophagy-suppressive phosphoinositide 3-kinase (PI3K) signaling pathway. In yeast, Vps18 and other members of the HOPS complex have been found essential for autophagic degradation. In Drosophila, the Vps18 homologue Deep orange (Dor) has previously been shown to mediate fusion of multivesicular endosomes with lysosomes. A requirement of Dor for ecdysone-mediated chromosome puffing has also been reported. In the present report, we have tested the hypothesis that Dor may control programmed autophagy at the level of ecdysone signaling as well as by mediating autophagosome-to-lysosome fusion. We show that dor mutants are defective in programmed autophagy and provide evidence that autophagy is blocked at two levels. First, PI3K activity was not down-regulated correctly in dor larvae, which correlated with a decrease in ecdysone reporter activity. The down-regulation of PI3K activity was restored by feeding ecdysone to the mutant larvae. Second, neither exogenous ecdysone nor overexpression of PTEN, a silencer of PI3K signaling, restored fusion of autophagosomes with lysosomes in the fat body of dor mutants. These results indicate that Dor controls autophagy indirectly, via ecdysone signaling, as well as directly, via autolysosomal fusion.     

Prevalence, Persistence, and Molecular Characterization of Glycopeptide-Resistant Enterococci in Norwegian Poultry and Poultry Farmers 3 to 8 Years after the Ban on Avoparcin

Environmental reservoirs of glycopeptide-resistant enterococci (GRE) in Norway have been linked to former growth promoting use of the glycopeptide avoparcin in poultry production. We have examined the prevalence of fecal GRE in poultry and poultry farmers 3 to 8 years after the Norwegian avoparcin ban in 1995 and performed molecular analyses of the GRE population. Fecal samples from poultry farmers and their flocks on 29 previously avoparcin-exposed farms were collected on five occasions during the study period (1998 to 2003). All flocks (100%) were GRE positive in 1998. Throughout the study period, 78.5% of the poultry samples were GRE positive. Glycopeptide-resistant Enterococcus faecium (GREF) was isolated from 27.6% of the farmer samples in 1998 and from 27.8% of the samples collected between 1998 and 2003. The prevalence of fecal GRE in poultry declined significantly during the study period, but prevalence in samples from the farmers did not decline. PCR analysis revealed a specific Tn1546-plasmid junction fragment in 93.9% of E. faecium isolates. A putative postsegregation killing (PSK) system linked to Tn1546 was detected in 97.1% of the isolates examined. Multilocus sequence typing of glycopeptide-susceptible (n = 10) and -resistant (n = 10) E. faecium isolates from humans (n = 10) and poultry (n = 10) on two farms displayed 17 different sequence types. The study confirms the continuing persistence of a widespread common plasmid-mediated vanA-pRE25-PSK element within a heterogeneous GRE population on Norwegian poultry farms 8 years after the avoparcin ban. Moreover, it suggests an important role of PSK systems in the maintenance of antimicrobial resistance determinants in reservoirs without apparent antimicrobial selection.     

Vascular dysfunction produced by hyperhomocysteinemia is more severe in the presence of low folate

Earlier we reported that dietary folate depletion causes hyperhomocysteinemia (HHcy) and arterial dysfunction in rats (Symons JD, Mullick AE, Ensunsa JL, Ma AA, and Rutledge JC. Arterioscler Thromb Vasc Biol 22: 772-780, 2002). Both HHcy and low folate (LF) are risk factors for cardiovascular disease. Therefore, the dysfunction we observed could have resulted from HHcy, LF, and/or their combination (HHcy + LF). We tested the hypothesis that HHcy-induced vascular dysfunction is more severe in the presence of LF. Four groups of rats consumed diets for approximately 10 wk that produced plasma homocysteine (microM) and liver folate (microg folate/g liver) concentrations, respectively, of 7 +/- 1 and 15 +/- 1 (Control; Con; n = 16), 17 +/- 2 and 15 +/- 2 (HHcy; n = 17), 10 +/- 1 and 8 +/- 1 (LF; n = 14), and 21 +/- 2 and 8 +/- 1 (HHcy + LF; n = 18). We observed that maximal ACh-evoked vasorelaxation was greatest in aortas and mesenteric arteries from Con rats vs. all groups. While the extent of dysfunction was similar between LF and HHcy animals, it was less severe compared with arteries from HHcy + LF rats. Maximal ACh-evoked vasorelaxation in coronary arteries was not different between Con and LF rats, but both were greater than HHcy + LF animals. In segments of aortas, 1) ACh-evoked vasorelaxation was similar among groups after incubation with the nonenzymatic intracellular O2(-) scavenger Tiron, 2) vascular O2(-) estimated using dihydroethidium staining was greatest in HHcy + LF vs. all groups, and 3) tension development in response to nitric oxide (NO) synthase inhibition was greatest in Con vs. all other groups. We conclude that HHcy + LF evokes greater dysfunction than either HHcy alone (aortas, mesentery) or LF alone (aortas, mesentery, coronary), likely by producing more O2(-) within the vasculature and thereby reducing NO bioavailability.