Heparin stimulates the collagen synthesis in mineralized cultures of the osteoblast-like cell line, MC3T3-E1.

We found that heparin induced an increase in collagen synthesis in MC3T3-E1 cells cultured for 15 and 30 days. Northern blots showed that the effect of heparin on the collagen synthesis was mediated through the mRNA expression of type I collagen (day 15). Although heparin stimulated collagen synthesis over and above that stimulated by the transforming growth factor (TGF)-beta, heparin did not stimulate TGF-beta binding.

Cloning of the gene encoding a glycylprolyl aminopeptidase from Porphyromonas gingivalis.

A genomic library of Porphyromonas gingivalis 381 was constructed in the cosmid vector pHC79. A clone, pSN1, was identified by the expression of glycylprolyl-naphthylamide hydrolysing activity. The DNA insert contained within the cosmid pSN1 was subcloned into the plasmid vector pBR328 to create the recombinant plasmid pSN11 containing a 2.

Stimulation by interleukin-1 of interleukin-6 production by human periodontal ligament cells.

Interleukin-1(IL-1), a cytokine present in the gingiva and crevicular fluid of patients with periodontitis and in the periodontal ligament (PDL) of experimentally moved teeth, has multiple biological activities, including the ability to elicit bone resorption. Interleukin-6, also found in the gingiva of patients with periodontitis, may induce osteoclastic bone resorption through an effect on osteoclastogenesis. Here IL-6 production and its gene expression in response to recombinant IL-1 beta were examined in primary cultures of PDL cells.

Role of Porphyromonas gingivalis 40-kDa outer membrane protein in the aggregation of P. gingivalis vesicles and Actinomyces viscosus.

Porphyromonas gingivalis, an important pathogen in periodontitis, produces extracellular vesicles that aggregate with Actinomyces viscosus cells. A 40-kDa outer membrane protein (OMP)-coding gene from P. gingivalis was cloned and the protein was found to be localized in these vesicles.

Suppressive effect of soluble factor(s) derived from Prevotella loescheii ATCC 15930 on proliferation of human lymphocytes.

Soluble sonic extracts of Prevotella loescheii caused a dose-dependent inhibition of human peripheral blood lymphocyte proliferation by mitogen and of the proliferation of a leukemic cell line, BALL-1, when assessed by DNA synthesis (3H-thymidine incorporation). RNA (3H-uridine incorporation) and protein (3H-leucine incorporation) synthesis were similarly altered after exposure to the extract. There was no effect on cell viability as measured by either trypan blue exclusion or extracellular release of the cytoplasmic enzyme lactate dehydrogenase.

Gene cloning of Porphyromonas gingivalis specific antigens recognized by serum of adult periodontitis patient.

1. Porphyromonas gingivalis is believed an important pathogen of adult periodontitis. A gene library of P.

Purification of sufficiently gamma-carboxylated recombinant protein C and its derivatives. Calcium-dependent affinity shift in immunoaffinity and ion-exchange chromatography.

Protein C, which is an important anti-thrombotic factor in the blood coagulation cascade, undergoes several post-translational modifications. gamma-Carboxylation on nine glutamic acid residues at the N-terminal region of the light chain [gamma-carboxylated glutamic acid (Gla) domain] is considered to be critical for full anti-clotting activity. It is also known that when recombinant protein C is expressed in animal cells this particular modification is often lost.

Cardioprotective effect of pindolol in ischemic-reperfused isolated rat hearts.

The effects of pindolol and timolol on ischemia reperfusion damage were studied in isolated working rat hearts. Ischemia (15 min) decreased the mechanical function and the energy state, and increased the tissue levels of free fatty acids (FFA). During reperfusion (20 min), the mechanical function did not recover, but the energy state recovered incompletely, whereas FFA increased further.

Positive inotropic and negative chronotropic effects of (-)-cis-diltiazem in rat isolated atria.

1. The cardiovascular effects of (-)-cis-diltiazem, an optical isomer of diltiazem, were studied in the isolated atrium and aortic strip. (-)-cis-Diltiazem (30 microM or more) increased the developed tension of the rat left atrium, while (+)-cis-diltiazem (1 microM or more) decreased it.

Role of oxygen radicals in canine myocardial metabolic derangement during regional ischemia.

To clarify the role of oxygen radicals in the development of myocardial injury during ischemia, production of lipid peroxides mediated by oxygen radicals was determined in in vivo dogs subjected to regional ischemia and reperfusion. Myocardial injury was assessed by derangement in energy and carbohydrate metabolism caused by ischemia and reperfusion. The production of lipid peroxides mediated by oxygen radicals considerably increased not only during reperfusion after ischemia but also during ischemia.

Attenuation of ischemia-induced regional myocardial acidosis by LP-805, a newly developed vasodilator, in dogs.

The effect of 8-tert-butyl-6,7-dihydropyrrolo[3,2-e]-5-methyl- pyrazolo[1,5-a]pyrimidine-3-carbonitrile (LP-805), a newly developed vasodilator, on myocardial acidosis induced by ischemia was studied in anesthetized open-chest dogs. Ischemia was induced by partially occluding the left anterior descending coronary artery. The coronary flow was artificially reduced to about 1/3 of the original flow.

Effects of thoracic epidural anesthesia on myocardial pH and metabolism during ischemia.

The effect of thoracic epidural anesthesia (TEA) on the ischemic myocardium was examined in open-chest dogs anesthetized intravenously. Ischemia induced by brief coronary artery occlusion caused an elevation of the ST segment in epicardial ECG and a reduction in myocardial pH and contractile force. TEA with 0.

The effect of pimobendan on myocardial mechanical function and metabolism in dogs: comparison with dobutamine.

The effect of pimobendan, a newly developed cardiotonic agent, on myocardial mechanical function and energy metabolism has been examined in the dog heart, and compared with that of dobutamine. Either saline, vehicle for pimobendan, dobutamine (0.3 and 1 microgram kg-1), or pimobendan (0.

Effects of human superoxide dismutase on ischemic and reperfused myocardium in isolated perfused rat heart.

The effect of human superoxide dismutase (h-SOD) on the ischemic heart was studied in the isolated perfused working rat heart. Myocardial mechanical function expressed as pressure-rate product decreased and completely stopped within 5 min after the onset of global ischemia, and never recovered after reperfusion following 20 min of ischemia. In the ischemic myocardium, the levels of ATP, ADP, and creatine phosphate decreased, and those of AMP and lactate increased.

The inhibitory effect of bevantolol on the accumulation of non-esterified fatty acids during ischaemia in the dog heart in situ.

1. The left anterior descending coronary artery (LAD) was completely ligated for 90 min (i.e.

Effect of NCO-700, an inhibitor of protease, on lysosomal rupture in the ischemic myocardium.

The effect of NCO-700 (1), a protease inhibitor, on subcellular distribution of lysosomal enzymes was studied in the ischemic perfused rat heart. Ischemia was induced by lowering the afterload pressure of the working heart preparation. The subcellular distribution of lysosomal enzymes was estimated by the ratio of the activities of cathepsin D, beta,N-acetylglucosaminidase, and acid phosphatase in the cytoplasm to the total enzyme activities.

Vacuoles and vesicles in the rat junctional epithelium: a study with serial ultrathin sections.

To investigate whether the electron-lucent structures resembling vesicles and vacuoles in the rat molar junctional epithelium (JE) are in fact intracellular or extracellular, a study using serial ultrathin sections was carried out. In one series of experiments, the animals were not treated before the tissues were conventionally fixed; in another, anesthetized animals were administered horseradish peroxidase 20 min before the tissues were fixed. A large number of electron-lucent structures resembling vesicles and vacuoles were detected in both the peripheral and central cytoplasm of the JE localized at enamel and connective tissue sites.

Effects of MCI-176, a new quinazolinone calcium antagonist, on myocardial energy and carbohydrate metabolism in ischemic dog hearts.

The effect of 2-(2,5-dimethoxyphenylmethyl)-3-(2-dimethylaminoethyl)- 6-isopropoxy-4(3H)-quinazolinone hydrochloride (MCI-176), a calcium antagonist, on ischemic myocardial metabolism was studied in dog hearts subjected to an occlusion of the left anterior descending coronary artery (LAD) for 3 or 30 min. MCI-176 (0.03 or 0.

An ultrastructural study of the pocket epithelium in rats.

The purpose of the present study was to elucidate the biological characteristics of pocket epithelium in experimental periodontitis. Elastic rubber was inserted between the first and second molars of the left maxilla in rats. The rubber was removed after one week, and the animals were sacrificed one week after the removal, i.

Purification and characterization of recombinant cell surface protein antigen A of Streptococcus sobrinus B13N.

1. It has been reported that immunization of rhesus monkeys with the surface protein antigen I/II from Streptococcus mutans significantly reduced dental caries. 2.

Purification and immunochemical characterization of a recombinant outer membrane protein from Bacteroides gingivalis.

1. Bacteroides gingivalis is thought to be one of the most virulent microorganisms in relation to adult periodontitis. A gene clone, MD125, is an Escherichia coli host which produces an outer membrane protein of B.

Platelet-activating factor stimulates production of prostaglandin E2 in murine osteoblast-like cell line MC3T3-E1.

We found that platelet-activating factor (PAF) stimulated the production of prostaglandin (PG) E2 in MC3T3-E1 cells in a time- and dose-dependent manner. 1.0 microM PAF gave a maximal stimulation of PGE2 production by MC3T3-E1 cells after a 4 hr PAF-treatment.

Both d-cis- and l-cis-diltiazem have anti-ischemic action in the isolated, perfused working rat heart.

The effect of diltiazem (d-cis-diltiazem) on the ischemic myocardium was compared with that of l-cis-diltiazem, an optical isomer having less potent calcium channel-blocking action, in the isolated, perfused working rat heart. Ischemia decreased mechanical function and tissue levels of ATP and creatine phosphate, and increased tissue levels of nonesterified fatty acids (NEFA), AMP and lactate. Reperfusion did not restore mechanical function, but restored incompletely the levels of metabolites (except NEFA) that had been altered by ischemia.

[Change of beta-actin gene expression during culture on osteoblast-like cell line, MC3T3-E1].

Multiple forms of actin (alpha, beta, and gamma-actin) have been found in a variety of mammalian cell lines and tissues by the use of high resolution, two-dimensional gel electrophoresis. alpha-Actin is found only in differentiated muscle cells, and its synthesis is induced during myogenesis in culture, beta- and gamma-actin isotypes are the principal components of microfilaments, structures believed to be involved in cellular morphogenesis, motility, and mitosis. Recent studies have shown that mammalian actin mRNA levels are modulated in response to changes in cell shape and during cell differentiation in vitro.

Dichloroacetate attenuates myocardial acidosis and metabolic changes induced by partial occlusion of the coronary artery in dogs.

The present study was undertaken to examine whether dichloroacetate, which inhibits pyruvate dehydrogenase kinase and, therefore, increases the activity of pyruvate dehydrogenase, attenuates myocardial acidosis and metabolic changes induced by coronary occlusion. In dogs anesthetized with pentobarbital, the left anterior descending coronary artery was incompletely occluded to reduce the left anterior descending flow to a half to one third of the original flow (partial occlusion) to produce myocardial (regional) ischemia. Partial occlusion was continued for 90 min, and a bolus injection of saline or dichloroacetate was made intravenously 30 min after the onset of occlusion.