DLA Class II Alleles Are Associated with Risk for Canine Symmetrical Lupoid Onychodystropy (SLO)

Symmetrical lupoid onychodystrophy (SLO) is an immune-mediated disease in dogs affecting the claws with a suggested autoimmune aethiology. Sequence-based genotyping of the polymorphic exon 2 from DLA-DRB1, -DQA1, and -DQB1 class II loci were performed in a total of 98 SLO Gordon setter cases and 98 healthy controls. A risk haplotype (DRB1*01801/DQA1*00101/DQB1*00802) was present in 53% of cases and 34% of controls and conferred an elevated risk of developing SLO with an odds ratio (OR) of 2.1. When dogs homozygous for the risk haplotype were compared to all dogs not carrying the haplotype the OR was 5.4. However, a stronger protective haplotype (DRB1*02001/DQA1*00401/DQB1*01303, OR = 0.03, 1/OR = 33) was present in 16.8% of controls, but only in a single case (0.5%). The effect of the protective haplotype was clearly stronger than the risk haplotype, since 11.2% of the controls were heterozygous for the risk and protective haplotypes, whereas this combination was absent from cases. When the dogs with the protective haplotype were excluded, an OR of 2.5 was obtained when dogs homozygous for the risk haplotype were compared to those heterozygous for the risk haplotype, suggesting a co-dominant effect of the risk haplotype. In smaller sample sizes of the bearded collie and giant schnauzer breeds we found the same or similar haplotypes, sharing the same DQA1 allele, over-represented among the cases suggesting that the risk is associated primarily with DLA-DQ. We obtained conclusive results that DLA class II is significantly associated with risk of developing SLO in Gordon setters, thus supporting that SLO is an immune-mediated disease. Further studies of SLO in dogs may provide important insight into immune privilege of the nail apparatus and also knowledge about a number of inflammatory disorders of the nail apparatus like lichen planus, psoriasis, alopecia areata and onycholysis.     

Crystal structure at 87 K of sodium α-l-guluronate dihydrate

X-Ray data collected at 87 K showed crystals of sodium α-l-guluronate dihydrate (C₆H₉O₇Na · 2 H₂O) to be orthorhombic, P2₁2₁2₁ with a = 7.591(2), b = 18.884(5), c = 6.842(2) Å, and Z = 4. The structure was solved by direct methods, and full-matrix least-squares refinement based on 1587 F_o yielded R = 0.043 and R_w = 0.033. The structure analysis indicates partial anomeric disorder with α:β ～90:10. The guluronate ring has the ¹C₄(l) conformation. Sodium binds two translation-equivalent guluronate units and one water molecule in a primary five-fold coordination. The complexing oxygen functions, which include all axial hydroxyl groups and one carboxylate oxygen atom in the guluronate ring, describe a distorted trigonal bipyramid. A prominent feature of the crystal structure is the stacks of sodium atoms and guluronate residues in alternating sequence along the c axis. The stacks are held together by an intricate system of hydrogen bonds involving all oxygen atoms in the structure. The water molecules play an important role in this system both as hydrogen donors and acceptors.     

Spatiotemporal Resolution of Rab9 and CI‐MPR Dynamics in the Endocytic Pathway

Rab9 is a small GTPase that localizes to the trans‐Golgi Network (TGN) and late endosomes. Its main function has long been connected to the recycling of mannose‐6‐phosphate receptors (MPRs). However, recent studies link Rab9 also to autophagy and lysosome biogenesis. In this paper, using confocal imaging, we characterize for the first time the live dynamics of the Rab9 constitutively active mutant, Rab9Q66L. We find that it localizes predominantly to late endosomes and that its expression in HeLa cells disperses TGN46 and cation‐independent (CI‐MPR) away from the Golgi yet, has no effect on the retrograde transport of CI‐MPR. We also show that CI‐MPR and Rab9 enter the endosomal pathway together at the transition stage between early, Rab5‐positive, and late, Rab7a‐positive, endosomes. CI‐MPR localizes transiently to separate domains on these endosomes, where vesicles carrying CI‐MPR attach and detach within seconds. Taken together, our results demonstrate that Rab9 mediates the delivery of CI‐MPR to the endosomal pathway, entering the maturing endosome at the early‐to‐late transition.      

Effects of interferon alpha on human osteoprogenitor cell growth and differentiation in vitro.

The specific effects of interferon alpha (IFNalpha), on the differentiation pathways of human osteogenic cells are not known. The aim of this study was to investigate possible effects of IFNalpha on osteogenic development by investigating cell differentiation, colony formation (colony forming unit-fibroblastic, CFU-F), cell proliferation, and gene expression, in particular bone morphogenetic protein (BMP) expression, of human bone marrow osteoprogenitor cells. Human bone marrow fibroblasts were cultured with or without the addition of IFNalpha (5-1,000 IU/ml) in the presence and absence of dexamethasone (10 nM) and ascorbate (100 microM), which are agents known to affect osteogenic differentiation. IFNalpha produced a significant dose-dependent inhibition of cell proliferation and alkaline phosphatase specific activity at concentrations as low as 50 IU/ml. IFNalpha (50-1,000 IU/ml) inhibited the stimulation of alkaline phosphatase specific activity induced by ascorbate and dexamethasone. Examination of CFU-F showed dose- and time-dependent inhibitions of colony formation and reductions in both colony size and alkaline phosphatase-positive CFU-F colonies particularly at earlier times. Reactivity with an antibody specific for osteoprogenitors (HOP-26), was reduced in IFNalpha-treated cultures. Northern blot analysis showed a significant dose-dependent up-regulation of BMP-2 mRNA, estrogen receptor alpha mRNA and osteocalcin mRNA expression in ascorbate/dexamethasone cultures. In contrast, IFNalpha significantly inhibited BMP-2 mRNA expression in the absence of ascorbate and dexamethasone. In conclusion, IFNalpha inhibits human osteoprogenitor cell proliferation, CFU- F formation, HOP-26 expression, and alkaline phosphatase specific activity and modulates BMP-2 gene expression. These results suggest a role for IFNalpha in local bone turnover through the specific and direct modulation of osteoprogenitor proliferation and differentiation.     

Test-retest reliability of three different countermovement jumping tests

In studies of physical performance comprising muscle strength and power, a vertical jump is a test method that frequently is used. It is important to have access to accurate measuring tools providing data with high reproducibility. Studies have shown that body composition also may play an important part in physical performance. The purpose of this study was to determine test-retest reliability for 3 different kinds of vertical jumps and to correlate jump height with body composition. Thirty-four normally trained subjects (women n = 17) between 18 and 25 years participated. Test-retest, on 3 kinds of vertical jumps, was performed with a median of 7 days between jumps. Methods used were a countermovement jump (CMJ) on a contact mat, with and without arm swing, and an Abalakow jump (AJ) using measuring tape, with arm swing. Body composition was assessed with the use of bioelectric impedance analysis. The results showed that high intraclass correlation coefficients (ICCs) were observed between testing occasions for all 3 vertical jumps (ICC between 0.48 and 0.88). The AJ in women presented the lowest ICC. Also the correlation between CMJ and AJ was high (rs = 0.88). Moderate-to-high correlations could be shown between body composition and CMJ in women (rs = -0.57-0.76). In conclusion, very high test-retest reliability for CMJ on a contact mat was found. For the AJ using a measuring tape, ICC were overall high, but a moderate nonsignificant ICC were found in women, indicating poor reproducibility. The data from the CMJ and AJ may be compared if approximately 25% of the AJ value is subtracted. In practice, this means that vertical jump tests have high reproducibility and can be used as measures of power development.     

Template-directed supramolecular assembly of a new type of nanoporous peptide-based material.

Dipeptides with two hydrophobic residues are known to often form crystals with hydrophobic or hydrophilic channels. One of the exceptions is Leu-Ile, which has been previously shown to crystallize as a nonporous 0.75 hydrate in the hexagonal space group P6(1) with Z' = 4. We have now found that in the presence of D-Leu, a second pseudopolymorph of the dipeptide is formed. The crystal structure has hydrophilic, water-filled channels with an irregular cross section of approximately 5.5 x 3.5 A and constitutes the first example of a crystal packing arrangement where the direction of the channels is perpendicular to the direction of hydrophobic stacking of side chains rather than parallel as in all other porous peptide-based materials.     

Proteomic identification of secreted proteins from human skeletal muscle cells and expression in response to strength training

Regular physical activity protects against several types of diseases. This may involve altered secretion of signaling proteins from skeletal muscle. Our aim was to identify the most abundantly secreted proteins in cultures of human skeletal muscle cells and to monitor their expression in muscles of strength-training individuals. A total of 236 proteins were detected by proteome analysis in medium conditioned by cultured human myotubes, which was narrowed down to identification of 18 classically secreted proteins expressed in skeletal muscle, using the SignalP 3.0 and Human Genome Expression Profile databases together with a published mRNA-based reconstruction of the human skeletal muscle secretome. For 17 of the secreted proteins, expression was confirmed at the mRNA level in cultured human myotubes as well as in biopsies of human skeletal muscles. RT-PCR analyses showed that 15 of the secreted muscle proteins had significantly enhanced mRNA expression in m. vastus lateralis and/or m. trapezius after 11 wk of strength training among healthy volunteers. For example, secreted protein acidic and rich in cysteine, a secretory protein in the membrane fraction of skeletal muscle fibers, was increased 3- and 10-fold in m. vastus lateralis and m. trapezius, respectively. Identification of proteins secreted by skeletal muscle cells in vitro facilitated the discovery of novel responses in skeletal muscles of strength-training individuals.