Biochemical studies of male gametogenesis in liliaceous plants.
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Biomedical subjects
Publications and source records attributed to Y Hotta.
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Tetrodotoxin, at concentrations up to 5 x 10(-6) gram per milliliter, has no effect on the spontaneous discharge in the smooth muscle of taenia coli. However, the spontaneous discharge is abolished by Mn(++) at a concentration of 0.5 millimole per liter. The contraction induced by immersing the muscle in isotonic KCl solution is also suppressed in the presence of Mn(++). Because Mn(++) is a specific suppressor of the spike induced by Ca(++) and tetrodotoxin is an inhibitor of the spike induced by Na(+), we suggest that Ca(++) is a charge carrier in the production of spike potential in the smooth muscle and that the entry of intervening Ca(++) through the membrane acts as a trigger for the contraction of smooth muscle.
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Thymidine may act as an inducer of thymidine kinase activity in cells of higher plants. The general response is demonstrable in a randomly developing cell population such as is found in germinating wheat embryos. If a synchronously developing cell population is studied, however, potentially inducible cells are found to be susceptible to the inductive effect of thymidine only during about 10 per cent of the G1 period, and close to the interval when thymidine kinase activity normally appears.
Young wheat roots were labeled with (32)P-inorganic phosphate. Following the labeling period, roots were homogenized in a sucrose medium and fractionated into nuclei, cytoplasmic particles (including proplastids and mitochondria), and a soluble fraction containing most of the microsomes. DNA prepared from the particles had a higher buoyant density than that from the nuclei and showed a marked loss in total label if the roots were exposed to non-radioactive medium for 48 hours prior to fractionation of the cells.
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The clinical significance of hepatocellular lipolysosomes in patients with various liver diseases was investigated. Of the 102 cases studied, lipolysosomes were found in 78 cases or 76%. In patients with fatty change of the liver, hepatocellular lipolysosomes were found in 62 of 68 or 91%. In patients without fatty change, lipolysosomes were found in 16 of 34 or 47%. The lipolysosome:lipid ratio in the hepatocytes ranged from 0 to 10%. No differences were found in the lipolysosome:lipid ratios of patients with acute hepatitis, cholestasis, fatty liver, chronic hepatitis, or cirrhosis. In chronic hepatitis and cirrhosis with associated fatty change, the ratio was correlated with the serum level of cholesterol, whereas in fatty liver the ratio was not correlated with any parameters of patient's age, etiologic factors, or serum level of cholesterol. The findings suggest that lysosomal function is less significant as a pathogenesis of fat infiltration but lipolysosomes increase in number as a sequence of cholesterol overload to the liver.
Plus/minus screening of the rabbit corneal cDNA library was performed using corneal and iris RNA as probes. Thirteen clones were isolated: three ferritin H-chains, a NADH-ubiquinone oxidoreductase B22 subunit, an alpha 1 type VIII collagen, a 25 KDa FKBP-506 binding protein (FKBP25), a thrombospondin 2, and six unknown clones. Although proteins translocated from these isolated mRNA are not corneal specific, they play an important role in the cornea. None of the isolated known mRNAs maps to chromosome 1, 16, or 20. These clones, thrombospondin excepted, were not observed in the high frequency clones in the profile of the aortic endothelial cDNA library.
We studied 133 Japanese patients with retinal dystrophies to detect peripherin/RDS (retinal degeneration slow) gene defects. The patients analyzed included 52 with autosomal dominant retinitis pigmentosa, 36 with autosomal recessive retinitis pigmentosa, 3 with simplex retinitis pigmentosa, 12 with cone-rod dystrophy, 5 with rod-cone dystrophy, 3 with vitelliform macular dystrophy (Best's disease), 4 with macular dystrophy, 2 with cone dystrophy, 2 with fundus flavimaculatus, 2 with fundus albipunctatus, and 12 with retinitis pigmentosa with macular degeneration as well as 40 unrelated normal persons. Three exons of the peripherin/RDS gene cut into 150-200 base-pair fragments were amplified by polymerase chain reaction and screened by single-strand conformation polymorphism. The DNA fragments with any suspected variations were directly sequenced. Eight point mutations were detected. Among them, two missense mutations at codons 304 and 338 result in an amino acid substitution of glutamine for glutamic acid and aspartic acid for glycine, respectively. However, they were not cosegregated with the diseases, and these mutations were also commonly found in normal controls. For these controls, the proportion of transversion from G to C at codon 304 (GAG-->CAG) and transition from G to A at codon 338 (GGC-->GAC) were 0.192 +/- 0.045 and 0.173 +/- 0.053, respectively. Our results suggest that a peripherin/RDS gene mutation might be rare in Japanese patients.
To characterize severe lattice corneal dystrophy, we analyzed the betaig-h3 gene, clinical features, histological findings, and genotype-phenotype correlation in an affected Japanese family. Deoxyribonucleic acid was extracted from leukocytes in 16 members (12 affected and 4 unaffected) of a Japanese family with lattice corneal dystrophy type I. Exon 4 of the betaig-h3 gene was amplified and analyzed using molecular biological methods. Clinical and pathological data were also collected. We found a heterozygous point mutation that causes the disease phenotype. It was a single base-pair transition leading to an amino acid substitution (CGC-->TGC, Arg124Cys). The phenotypic variation within families was not recognized. The affected members in the pedigree demonstrated severe visual disturbance in the third decade and required keratoplasty. Histopathological examination revealed amyloid deposits consisting of short and thin amyloid fibers and lattice corneal dystrophy type I. The heterozygous Arg124Cys mutation reported in Caucasian lattice corneal dystrophy caused severe lattice corneal dystrophy consisting of short and thin amyloid fibers in a Japanese family. Based on our study of many members of the family, we are able to construct the natural course of this disorder from its earliest clinical findings through its late manifestations.
Poly-L-lactides containing beta-alkyl alpha-malate-units were prepared by ring-opening copolymerizations of L-lactide with 3-(s)-[(benzyloxycarbonyl)methyl]- (BMD) and 3-(s)-[(dodecyloxycarbonyl)methyl]-1,4-dioxane-2,5-diones (DMD). The solution-cast films of these copolymers were alkali-treated to form a carboxyl-functionalized surface on which cell-binding Arg-Gly-Asp tripeptide (RGD) was immobilized with dicyclohexylcarbodiimide as coupling agent. For the copolymer of L-lactide and BMD the benzyl groups were removed by catalytic hydrogenolysis to obtain a fully carboxyl-functionalized copolymer (PLGM), and RGD was immobilized on the surface of its cast film. All the RGD-immobilized films thus prepared exhibited improved cell attachment compared with the original films. The cell attachment increased with increasing amount of immobilized RGD, which depended on the composition of the alpha-malate units in the copolymer. The RGD-immobilized PLGM films were degraded rapidly during the cell culture, while the RGD-immobilized films of the beta-alkyl alpha-malate-containing polymers survived the cell culture with little degradation. The rate of hydrolysis increased with increasing content of alpha-malate units for both series, depending on the structure of the protecting groups of the beta-carboxyl. These results suggest that the RGD-immobilized polymers could be a new class of functional bioresorbable polymer having improved cell-attachment and adjustable hydrolysis rate.