Anti-proliferative effect of 2-methoxyestradiol on cultured smooth muscle cells from rabbit aorta.

The proliferation of cultured smooth muscle cells from rabbit aorta was inhibited markedly by 2-methoxyestradiol, considerably by 2-hydroxyestradiol, and only slightly by 17 beta-estradiol. The effects at doses from 1 to 40 nmol/ml were cytostatic rather than cytocidal. 2-Methoxyestradiol could be useful to suppress excessive proliferation of arterial smooth muscle cells, and thus to prevent the progression of atherosclerosis.

Lipid peroxide levels and lipids content of serum lipoprotein fractions of pregnant subjects with or without pre-eclampsia.

Total serum lipids in normal pregnant subjects were significantly higher than those in non-pregnant subjects. This was due to an increase in the lipid content in all of the lipoprotein fractions, especially in VLDL. Serum lipids in pre-eclamptic subjects were higher than those in normal pregnant subjects. This was due to the increase in both VLDL and LDL. Serum lipid peroxide levels in pregnant subjects were significantly higher than those in non-pregnant subjects and further elevation of the levels was observed in pre-eclamptic subjects. The elevation of the peroxide levels in normal pregnant subjects was due to changes in all of the fractions of serum lipoproteins, especially in VLDL. The further elevation of peroxide in pre-eclamptic subjects resulted from an increase in HDL fraction.

Possible contributions of mastocytosis, apoptosis, and hydrolysis in pathophysiology of randomized skin flaps in humans and guinea pigs.

To understand better the pathophysiology of random skin flaps, randomized skin flaps of human (3 cases) and guinea pig (53 cases) were investigated. Proximal (normal), proximomedial (viable), mediodistal (between viable and necrotic parts), and distal (necrosis) locations of the skin flaps were biopsied. Lipid peroxidase, hydrolytic enzymes of cytosol (Ca(2+)-dependent cysteine protease: calpain), and lysosome (acid phosphatase) of skin were used as markers. Measurements were taken of the flap blood flow; the numbers of capillaries, postcapillary venules, pericapillary arterioles, leukocytes, and mast cells per unit square of dermis. Apoptotic cells were identified by specific staining. Flaps were sampled at postoperative weeks 1 and 3 (human) and hours 1 and 6, and days 1 to 7 (guinea pig). The values for normal skin were regarded as the control. Obstruction (by leukocytes) of venous microvessels, rather than arterial microvessels, was the major cause of temporary hypoxia in the proximomedial location, constant hypoxia (venous stasis) in the mediodistal location, and ischemia in the distal location. Increases in the number of mast cells (mastocytosis) and microvessels (angiogenesis) were significant only in the viable parts of the flaps. This phenomenon and the rate of blood flow increased with time in viable locations (guinea pig). Epidermal necrosis, dermal fibrosis, and apoptosis were evident mostly in the mediodistal location. Elevated levels of leukocytes, lipid peroxidase, acid phosphatase, and calpain, combined with necrotic changes, were seen mostly in the distal skin location. There is a strong possibility that the following factors are involved: lipid piroxidation and hydrolysis in necrosis of the distal flap location after ischemia; constant hypoxia in fibrosis and apoptosis in the mediodistal location; and initial or temporary hypoxia in mastocytosis-induced angiogenesis in the viable location. The results presented here indicate that guidelines for further investigations include combined suppression of leukotaxis, lipid peroxidase, and hydrolysis, or the application of mast cell growth factors in an effort to salvage the flap maximally.

Studies on myoinositol. Effects of caffeine and theophylline on incorporation of myoinositol into phosphatidylinositol in rat liver homogenate.

Both caffeine and theophylline, which were known to be potent inhibitors of cyclic-AMP phosphodiesterase, stimulated the incorporation of myoinositol into phosphatidylinositol in rat liver homogenate. However, cyclic-AMP had no effect. The effect of dibutyryl-cyclic-AMP differed with different concentrations. These results suggest that the stimulation cannot be explained by the increase in the amount of cyclic-AMP. This view was supported by the fact that papaverine, cyclic-AMP phosphodiesterase inhibitor, did not stimulate the incorporation and imidazole, the phosphodiesterase stimulator, did not inhibit the incorporation, and that adenylcyclase stimulators, epinephrine and glucagon, did not stimulate the incorporation.

[Three interesting pediatric cases with penicillin-resistant Streptococcus pneumoniae infection].

Since penicillin-resistant Streptococcus pneumoniae first recognized in 1967, the rate of penicillin-resistant strains has been increasing worldwide. There have been up to 50% from pediatric specimens in Japan. We reported three pediatric cases with penicillin G resistant Streptococcus pneumoniae infection to show some important clue from these cases for clinical practice against resistant pneumococcal infection. The first case was a typical acute mastoiditis, although we have experienced only masked mastoiditis recently. The second case was meningitis with septicemia, which did not show any abnormality in the first obtained cerebrospinal fluid. The third case was recurrent bronchitis in a child with cerebral palsy. The minimum inhibition concentrations of these isolated strains were 0.25 microgram/ml in the second case an 2.0 microgram/ml in the first and third cases.

Isoelectric focusing studies of human red cell esterase D: evidence for polymorphic occurrence of a new allele EsD7 in Japanese.

The isoelectric focusing study of esterase D in Japanese revealed evidence of a new polymorphic allele (EsD7) which is difficult to find by conventional starch gel electrophoresis only. A comparison with the occurrence of a subdivision of EsD2 in Caucasians (EsD5) suggests a remarkable difference in allele distribution of esterase D among races. Quantitative analysis showed a relatively low value of enzyme activity for this new allele. It is therefore emphasized that in addition to conventional electrophoresis, enzyme assay and further detection by isoelectric focusing are essential in analyzing the esterase D system.

Quantitative variations in polymorphic types of human red cell esterase D.

Human red cell esterase D was analysed quantitatively in Japanese samples. Among the three esterase D phenotypes there was a significant difference in catalytic activity, with an approximate ratio of 7:5:3 for EsD 1, EsD 2-1 and EsD 2. With a few inter-type individuals, the distribution of their activity values was plotted in a tri-modal curve. These results suggest that the esterase D system in human red cells is qualitatively and even quantitatively controlled by genes.

A rare phenotype of phosphoglucomutase-2 first detected in Mongoloids.

In screening of the PGM system by isoelectric focusing a heterozygote with a rare allele of PGM2 was detected. Family studies and family register checkings for the propositus showed evidence of hereditary occurrence of this allele, and all the ancestors of the propositus were Japanese. Although there have been no reports among Mongoloids, the variant found in the present study is likely to be the first case of a rare phenotype of PGM2 among such populations. In addition, isoelectric focusing data on 411 Japanese samples are given.

Purification of aromatic L-amino acid decarboxylase from bovine brain with a monoclonal antibody.

Aromatic L-amino acid decarboxylase was purified from bovine brain for the first time by affinity chromatography using a monoclonal antibody to the enzyme, and it was compared with the decarboxylase purified from bovine adrenal medulla by the same procedure. The monoclonal antibody was produced from a hybridoma established for the enzyme highly purified from bovine adrenal medulla. The Mr values of brain and adrenal-medulla enzyme were both estimated to be approx. 100,000 by gel-permeation chromatography. SDS/polyacrylamide-gel electrophoresis revealed a single band with an apparent Mr of 50,000. Western immunoblot analysis showed that the antibody recognized each enzyme. With regard to substrate specificity, pH-dependence and effect of pyridoxal 5'-phosphate as a cofactor, both enzymes were similar.

Isoelectric focusing studies of human red cell PGM1 in Japanese, with special reference to the characterization of PGM17.

The distribution of phosphoglucomutase (PGM1) subtypes in human red cells was determined by isoelectric focusing in 218 Japanese samples. Nine common phenotypes were observed corresponding to the following frequencies of the four alleles at the PGM1 locus: PGM11+ 0.6560, PGM11- 0.1170, PGM12+ 0.1674 and PGM12- 0.0505. In addition, a characterization of the PGM17 allele was performed. Our results obtained in the present study revealed the possibility that the PGM17 allele may be differentiated in the two alleles of PGM17+ and PGM17- through an investigation of isoelectric focusing.

Genetic studies of red cell glutamic-pyruvic transaminase in some Japanese populations.

Comparative data on the allelic distribution of human red cell GPT were presented for inhabitants of isolated communities and individuals in the general population of Japan. In addition, a quantitative enzyme assay of the different phenotypes was carried out on some of the samples tested. The existence of two heterozygotes with a silent allele, Gpt0, appeared to be confirmed by pedigree analysis.

Glycated protein-iron chelate increases lipid peroxide level in cultured aortic endothelial and smooth muscle cells.

Formation of an iron chelate of glycated protein was demonstrated by the appearance of an absorption peak at approximately 270 nm after mixing glycated bovine serum albumin with FeCl3. This peak disappeared and a new peak appeared at approximately 420 nm to form an isosbestic point at approximately 340 nm by the addition of deferoxamine mesylate, an iron-chelating agent, to the mixture, thus confirming the formation of the iron chelate of the glycated protein in the mixture. The lipid peroxide level was increased markedly in endothelial cells and slightly in smooth muscle cells from bovine aorta incubated in the medium containing glycated fetal bovine serum-iron chelate. Morphological observation by phase-contrast microscopy and scanning electron microscopy revealed that the glycated fetal bovine serum-iron chelate caused intense damage to the endothelial cells. These results indicate that glycated protein-iron chelate provokes lipid peroxidation, which explains at least in part the mechanism of atherogenesis found in diabetic patients.

Age-dependent changes in lipid peroxide levels in the lipoprotein fractions of human serum.

Lipid peroxide levels in serum lipoprotein fractions of young and old individuals, who had no significant disease, were studied. The lipid peroxide level, amount of lipids, and the ratio of the former to the latter were significantly elevated in the low density lipoprotein fraction of older adults. This indicates that the elevation of the serum lipid peroxide levels in older adults is mainly due to increased lipid peroxide level in the low density lipoprotein fraction.