Sequence polymorphisms in porcine homologs of murine coat colour-related genes.

Herein, we report the variability among 57 porcine homologs of murine coat colour-related genes. We identified single nucleotide polymorphisms (SNPs) and insertions/deletions (InDels) within 44 expressed gene sequences by aligning eight pig complementary DNA (cDNA) samples. The sequence alignment revealed a total of 485 SNPs and 15 InDels. The polymorphisms were then validated by performing matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) with reference DNA samples obtained from 384 porcine individuals. Of the 384 individuals, three parents of the experimental F(2) family were included to detect polymorphisms between them for linkage mapping. We also genotyped previously reported polymorphisms of 12 genes, and one SNP each in three genes that were detected by performing a BLAST search of the Trace database. A total of 211 SNPs and three InDels were successfully genotyped from our porcine DNA panel. We detected SNPs in 33 of the 44 genes among the parents of an experimental F(2) family and then constructed a linkage map of the 33 genes for this family. The linkage assignment of each gene to the porcine chromosomes was consistent with the location of the BAC clone in the porcine genome and the corresponding gene sequence. We confirmed complete substitutions of EDNRB and MLPH in the Jinhua and Clawn miniature breeds, respectively. Furthermore, we identified polymorphic alleles exclusive to each pig group: 13 for Jinhua, two for Duroc, three for Meishan, four for the Japanese wild boar, one for the Clawn miniature pig and four for the Potbelly pig.

Three-dimensional analysis of pathological characteristics of a microaneurysm.

UNLABELLED: The etiology of Charcot-Bouchard microaneurysms (MAs), especially before rupture, has been unclear. CASE REPORT: A surgical specimen of an MA was removed from an acute intracerebral hemorrhage in a 69-year-old man. The MA was fixed with formalin and embedded in paraffin, and serial sections were cut and stained by the Azan and Elastica van Gieson methods and immunohistochemically. Three-dimensional reconstructive analysis was performed. RESULTS AND DISCUSSION: The MA presented as local enlargement of an artery with a maximal diameter of 840 microm, but was not ruptured. The wall of the MA consisted of thickened, fine collagen fibrils, which were found to be similar to adventitia immunohistochemically, but with no internal elastic lamina or medial smooth muscle layer. The lumen was empty, with no mural thrombi, but the inner wall endothelial cells had expanded from the normal and/or residual arterial wall, which suggested that the MA had been present for a long period of time. Because the bare wall of the MA consisted of thickened collagen fibrils, the MA was not ruptured, but the possibility of rupture in the future was deemed high, with the risk of intracerebral hemorrhage.

Immobilization of human thrombomodulin onto poly(ether urethane urea) for developing antithrombogenic blood-contacting materials.

Thrombomodulin (TM) is a newly described endothelial cell-associated protein that functions as a potent natural anticoagulant by converting thrombin from a procoagulant protease to an anticoagulant. In this study, focussing on the application of TM for biomedical materials, recombinant human TM (hTM) was immobilized onto the polymers for medical use, and the evaluation of their antithrombogenicity and the interaction with platelets were investigated. As the base polymer for immobilization reaction, poly(ether urethane urea) (PEUU), which was reported to have good blood compatibility, was used. hTM-immobilized PEUU showed superior antithrombogenic activity, such as the prolongation of plasma recalcification time and the inhibition of thrombin-induced platelet aggregation, though the amount of immobilized hTM was very small (i.e. less than 1 microgram/cm2). Platelet adhesions onto hTM-immobilized PEUU were not observed. These results show that the immobilization of hTM does not change the native good blood compatibility of PEUU, but provides excellent anticoagulant activity.

Immobilization of human thrombomodulin on glass beads and its anticoagulant activity.

Human thrombomodulin (TM) was for the first time immobilized on glass beads by the reaction between the carboxyl group of TM and the amino group of glass beads using water-soluble carbodiimide. Immobilized human TM exhibited both anticoagulant activity and inhibition of platelet aggregation of human blood.