Cardiac Restricted Overexpression of Membrane Type-1 Matrix Metalloproteinase Causes Adverse Myocardial Remodeling following Myocardial Infarction

The membrane type-1 matrix metalloproteinase (MT1-MMP) is a unique member of the MMP family, but induction patterns and consequences of MT1-MMP overexpression (MT1-MMPexp), in a left ventricular (LV) remodeling process such as myocardial infarction (MI), have not been explored. MT1-MMP promoter activity (murine luciferase reporter) increased 20-fold at 3 days and 50-fold at 14 days post-MI. MI was then induced in mice with cardiac restricted MT1-MMPexp (n = 58) and wild type (WT, n = 60). Post-MI survival was reduced (67% versus 46%, p < 0.05), and LV ejection fraction was lower in the post-MI MT1-MMPexp mice compared with WT (41 ± 2 versus 32 ± 2%,p < 0.05). In the post-MI MT1-MMPexp mice, LV myocardial MMP activity, as assessed by radiotracer uptake, and MT1-MMP-specific proteolytic activity using a specific fluorogenic assay were both increased by 2-fold. LV collagen content was increased by nearly 2-fold in the post-MI MT1-MMPexp compared with WT. Using a validated fluorogenic construct, it was discovered that MT1-MMP proteolytically processed the pro-fibrotic molecule, latency-associated transforming growth factor-1 binding protein (LTBP-1), and MT1-MMP-specific LTBP-1 proteolytic activity was increased by 4-fold in the post-MI MT1-MMPexp group. Early and persistent MT1-MMP promoter activity occurred post-MI, and increased myocardial MT1-MMP levels resulted in poor survival, worsening of LV function, and significant fibrosis. A molecular mechanism for the adverse LV matrix remodeling with MT1-MMP induction is increased processing of pro-fibrotic signaling molecules. Thus, a proteolytically diverse portfolio exists for MT1-MMP within the myocardium and likely plays a mechanistic role in adverse LV remodeling.     

Cyclic GMP-dependent and cyclic AMP-dependent protein kinases,protein kinase modulators and phosphodieterases in arteries and veins of dogs Distribution and effects of arteriovenous fistula and arterial occlusion

Possible involvement of cyclic GMP-dependent and cyclic AMP-dependent protein kinases, protein kinase modulators and cyclic nucleotide phosphodiesterases in functions of vascular tissues were investigated in the dog. All of the above activities, localized in the smooth muscle-rich inner layer of the blood vessels, were found to be higher in the arteries than in the veins. The peripheral arteries were disproportionately richer in cyclic GMP-dependent protein kinase (as indicated by high ratios of cyclic GMP-dependent to cyclic AMP-dependent protein kinase) than were the veins, with the exception of the pulmonary artery, an atypical arterial tissue exposed to low blood pressure. Interestingly, the protein kinase ratio for the aorta, an artery with no significant role in blood pressure regulation, was not higher than that for the vena cava. Creation of femoral arteriovenous fistulae in the dogs led to preferential reductions in the cyclic GMP-dependent enzyme activity both in the proximal and distal arteries, whereas it was elevated in the stressed vein distal to the anastomotic site. The cyclic GMP-dependent enzyme was preferentially reduced in the saphenous artery distal to occlusion. Changes in the cyclic GMP-dependent enzyme activity appeared to precede gross atrophy or hypertrophy of the vessels. It is suggested that the vascular cyclic GMP-dependent protein kinase may be closely related to peripheral resistance and its regulation.     

Distribution and functional role of laminin during induction of the embryonic axis in the chick embryo

Laminin is a major glycoprotein of basement membranes and has been shown to promote cell adhesion, and movement of various nonepithelial cells and tumour cells. Using antibodies to laminin in paraffin sections and cultured embryos, we have studied the distribution of laminin and its involvement in the first morphogenetic events, beginning with the first extensive cellular migrations and interactions that result in the induction of the primitive streak (PS) and of the neural plate in the early chick embryo. Laminin immunogold labeling was not detected in the blastoderm at stage X. At stage XIII, laminin immunoreactivity was detected at the ventral surface of the epiblast and in the entire hypoblast. The intense labeling of the hypoblast indicated that these cells are active in laminin synthesis. Extracellular matrix (ECM) started accumulating as the first embryonic spaces were forming, before the morphogenetic movements of gastrulation were initiated. Immunogold labeling revealed a punctate pattern of laminin distribution in the ECM in the blastocoele, and in the space below the neural plate. Laminin, which is a multidomain molecule known to interact with other molecules of the ECM and with the cell surface, could serve as the scaffold for highly specific contact points of migrating cells and for the folding of epithelial sheets during this time in the developing embryo. We incubated blastoderms at stages X and XIII with laminin antibodies (1:30 dilution) for 4 h, then cultured the blastoderms further in plain egg albumin. The laminin antibodies did not interfere with triggering of PS cell movements, but perturbed the normal migration pattern of these cells. A normal PS did not form and, as a consequence, the embryonic axis was not induced.(ABSTRACT TRUNCATED AT 250 WORDS)     

Squid glycoproteins with structural similarities to Thy-1 and Ly-6 antigens

In an attempt to identify invertebrate homologs of Thy-1 antigen, the optic and central nervous tissue of squid was solubilized in deoxycholate and fractionated by lentil lectin affinity chromatography and gel filtration to yield small abundant glycoproteins. Material with biochemical similarities to Thy-1 was found and shown to consist of two glycoproteins that were ultimately purified using monoclonal antibody affinity columns. Both glycoproteins were sequenced to yield sequences of 84 residues for Sgp-1 and 92 residues for Sgp-2. The sequences were analyzed for similarities to Thy-1 and other Ig-related sequences, and Sgp-1 showed some similarities that were > 3 standard deviation units away from mean random scores when tested with the ALIGN program. However, the sequence patterns were not typical of Ig-related domains and the relationship of Sgp-1 to the Ig superfamily remains problematical. Sgp-2 showed no relationship to the Ig superfamily, but similarities to Ly-6 antigen sequences were noted that are in accord with an evolutionary relationship. The similarities included ten Cys residues in each sequence of which eight were matched in the best alignment given by the ALIGN program. Chemical evidence was obtained for glycophospholipid tails at the COOH-termini of Sgp-1 and Sgp-2 as is the case for Thy-1 and Ly-6 antigens.Abbreviations used in this paper GPL glycophospholipid - mAb monoclonal antibody - SDS-PAGE sodium dodecyl sulfate-polyacrylamide gel electrophoresis     

Susceptibility of laboratory-reared northern fowl mites,Ornithonyssus sylviarum (Acari: Macronyssidae), to selected acaricides

Toxicity was determined for 15 acaricides against a laboratory strain of northern fowl mites,Ornithonyssus sylviarum (Canestrini and Fanzago). Adult females were exposed to residues on filter paper for 24 h. Three organophosphorous compounds (monocrotophos, cythioate, and famphur) were more toxic to the northern fowl mite than was carbaryl, the most commonly used pesticide in the poultry industry. The other tested compounds were less toxic to the mite than was carbaryl. Four of these, not used previously for northern fowl mite control, had low LC₅₀'s for northern fowl mites: aldicarb (0.46); pirimiphos-methyl (0.73); exo, exo-2,8-dichloro-4-thiatricyclo [3.2.1.0^3.6]octane-4-oxide (AI3-63182) (0.87); and diazinon (2.48).This article reports the results of research only. Mention of a pesticide or proprietary product does not constitute an endorsement or a recommendation of its use by U.S.D.A.     

Application of bromodeoxyuridine incorporation measurements to the determination of cell distribution within the S phase of the cell cycle

Flow cytofluorimetric measurement of incorporated bromodeoxyuridine, using a double-stained cell population, allows the determination of the distribution of cells along the cell cycle. We have developed a simple computer program for the direct treatment of 64 x 64 channel histograms. This analysis appears to provide interesting data about the distribution of cells in the various phases of the cell cycle, namely the S phase. Two examples have been chosen to illustrate possible fields for the application of such a program. Comparison of two cell lines such as friend murine erythroleukemia cells (MELC) and fibroblasts FR3T3 cells has shown that this analysis can be used for cell-cycle characterization of a given cell line. The program also allows the differential analysis of cell distribution along the cell cycle as a function of a given parameter. This possibility has been applied to study the variation of cell-cycle parameters as a function of the time of induced differentiation of MELC and reveals changes in the distribution of the cells along the various phases of the cell cycle, namely in the S phase.     

Lipid peroxidation is a consequence of elicitor activity

Elicitor-active preparations from the fungal pathogen of bean Colletotrichum lindemuthianum stimulated the accumulation of products characteristic of lipid peroxidation in treated bean tissues. Bean suspension cells treated with crude and purified elicitors accumulated `lipofuscin-like pigment' (LEP) and malondialdehyde. The accumulation of LFP after about 6 h of treatment coincided with the onset of visible browning and production of the bean phytoalexins kievitone, phaseollin, and phaseollinisoflavan. The induction of phytoalexins and accumulation of LFP were also triggered by treatments with generators of activated oxygen species, xanthine:xanthine oxidase and Fe:ethylenediaminedi-o-hydroxyphenylacetic acid. These data suggest that generation of active oxygen species may be involved in lipid peroxidation triggered by elicitors.     

Mouse Chromosome 7

Chair of Committee for Mouse Chromosome 7