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T Iino

Publications and source records attributed to T Iino.

At least 109 records · Page 6Linked to original sources

Role of atherosclerosis in spontaneous periodic contractions of isolated postmortem human coronary arteries.

The role of atherosclerosis in coronary artery spasm was studied by morphometrical examinations of isolated postmortem human coronary arteries that were used for the observation of spontaneous periodic contractions. Isometric tension of coronary ring segments from seventy-six cadavers was recorded in a bath containing physiological saline solution. According to the degree of atherosclerosis, the segments were classified into three groups: minimum intimal lesion (Group I), moderate intimal lesion (Group II) and severe atherosclerotic lesion (Group III). Results were analyzed by a chi-square test. The incidence of the spontaneous periodic contractions was 45% in the proximal portion. Of the segments in which the contractions occurred, 83% were in Group II. There were significant differences in the incidence between Group I and II (33% vs 80%; p less than 0.02) and Group II and III (80% vs 25%; p less than 0.001), respectively. According to the characteristics of the atherosclerosis, there were significant differences in incidence of the contractions between circumferential thickening and eccentric thickening in Group II (63% vs 28%; p less than 0.001). In conclusion, the spontaneous periodic contractions occur most frequently in the segments in which (1) the degree of atherosclerosis is moderate rather than minimum or severe and (2) the characteristics of atherosclerosis is circumferential thickening instead of eccentric thickening.

Adolescent↗

Mast cell-Ito cell pairings found in the Disse's spaces in the liver of the beagle dog.

Ito cells (fat-storing cells), containing a few lipid droplets, appeared in the Disse's spaces of the Beagle dog liver. They ranged in diameter from 0.5 to 5 micron. The present study revealed that mast cells were also distributed in the Disse's spaces, while they were encountered only in the interlobular connective tissue in other animals. Although mast cells as well as Ito cells were distributed solitarily in the Disse's spaces, pairing of both types of cells could be frequently observed. Direct contact between both cells was verified using both light and electron microscopy. Also encountered were some peculiar figures of certain Kupffer cells engulfing the cytoplasmic processes and secretory granules of mast cells. These findings indicate a previously unknown intimate relationship between the mast cell and other perisinusoidal cells in the dog liver: the Ito cell, Kupffer cell and endothelial cell.

Animals↗

On the nucleotide sequence recognized by a eukaryotic site-specific endonuclease, Endo.SceI from yeast.

Endo.SceI which is isolated from cells of Saccharomyces cerevisiae is a eukaryotic site-specific endonuclease active on double-stranded DNA. At each cleavage site, Endo.SceI cuts only a defined phosphodiester bond in each strand of the double helix. We compared nucleotide sequences around five cleavage sites for Endo.SceI using a computer. We could not find any common specific sequence consisting of five base pairs or more among them. However, we found a 26-base pair consensus sequence which included 15 conserved nucleotides, allowing any of the five sequences to include a few nucleotides deviated from the consensus sequence. The consensus sequence is 5'-CAn*PYnnAnnCYYGTTnnnPnYnnYA-3', where P, Y, n, and * denote purine, pyrimidine, any nucleotide, and the center of the cleavage site, respectively. The numbers of sites at which the consensus sequence appears in pBR322 DNA, phi X174 replicative form DNA, fd replicative form DNA, or SV40 DNA are close to those of the cleavage sites for Endo.SceI. We found that a 33-base pair fragment was efficiently cut at the defined phosphodiester bonds by Endo.SceI. This 33-base pair fragment included 25 base pairs out of the 26-base pair consensus sequence. The fragments in which a part of the consensus sequence was missing were not cut by Endo.SceI. These observations suggest that the consensus sequence described above is the major characteristic around the cleavage sites recognized by Endo.SceI and that the mode of recognition of cleavage sites by Endo.SceI is different from that by restriction endonucleases. We found homology between the consensus sequence for Endo.SceI and the sequences around the cleavage sites for two other site-specific endonucleases of S. cerevisiae: Endo.SceII and YZ-Endo which is involved in mating type switching.

Base Composition↗

The E. coli K-12 chromosome flanked by two IS10 sequences transposes.

Transposon are commonly found among prokaryotes and usually range up to 20 kilobases. In this study, we were interested to determine whether a larger DNA segment could transpose. We observed that the E. coli K-12 chromosome, 4,000 kilobases in size, when flanked by two IS10 sequences, could transpose to pACYC177 at a frequency of 10(-8) per cell per generation. We suggest that this transposition event occurs independently of the size and without duplication of the entire DNA sequence flanked by the IS10 elements.

Chromosomes, Bacterial↗

An essential gene for replication of the mini-F plasmid from origin I.

We constructed a series of defective mini-F plasmids, which have deletion(s) in the replication origin I and/or origin II, and their derivatives, which do not produce F3 protein, by insertion of the XhoI fragment of Tn5 into the XhoI site at 41.0 F (kilobases on the coordinate map of F-plasmid). Using these mutant mini-F plasmids, we found that F3 protein is essential for the replication of mini-F from origin I, but not from origin II.

Bacterial Proteins↗

Does Tn10 transpose via the cointegrate molecule?

It has been well established that Tn3 and its relatives transpose from one replicon to another by two successive reactions: formation of the cointegrate molecule and resolution from it. Whether or not the 9300 base pair tetracycline resistance transposon Tn10 transposes in the same manner as Tn3 was investigated by two methods. In the first method, lambda 55, a lambda phage carrying Tn10 was lysogenized in an Escherichia coli strain carrying a Tn10 insertion; the phage has a deletion in attP, hence it was lysogenized in a Tn10 sequence in the E. coli chromosome by reciprocal recombination. The chromosomal structure in these lysogens is equivalent to the Tn10-mediated cointegrate molecule of lambda and the E. coli chromosomal DNA. The stability of the cointegrate molecule was examined by measuring the rate of excision of lambda from the host chromosome, and was found to be stable, especially in a Rec- strain. Because of this stability, the cointegrate molecule should be accumulated if Tn10 transposes via the cointegrate molecule. Then, we examined the configuration of products made by transposition of Tn10 from lambda 55 to the E. coli chromosome. The cointegrate molecule was found in products of Tn10 transposition in a Rec+ strain at a frequency of 5% per Tn10 transposition, but this molecule could not be found in a Rec- strain. Since transposition of Tn10 was recA-independent, absence of the cointegrate molecule formed in a RecA- strain strongly suggested that the cointegrate molecule is not an obligatory intermediate of transposition of Tn10.(ABSTRACT TRUNCATED AT 250 WORDS)

Bacteriophage lambda↗

Tn501 insertion mutagenesis in Pseudomonas aeruginosa PAO.

Transposon insertion mutagenesis of the Pseudomonas aeruginosa PAO chromosome with Tn1 and Tn501 was carried out using a mutant plasmid of R68::Tn501 temperature-sensitive for replication and maintenance. This method consists of three steps. Firstly, the temperature-independent, drug-resistant clones were selected from the strain carrying this plasmid. In the temperature-independent clones, the plasmid was integrated into the chromosome by Tn1- or Tn501-mediated cointegrate formation. Secondly, such clones were cultivated at a permissive temperature to provoke the excision of the integrated plasmid from the chromosome. Excision occurred by the reciprocal recombination between the two copies of Tn1 or Tn501 flanking the integrated plasmid, leaving one Tn1 or Tn501 insertion on the chromosome. Thirdly, the excised plasmid was cured by cultivating these isolates at a non-permissive temperature without selection for the drug resistance. Using this method, we isolated 1 Tn1-induced and 43 Tn501-induced auxotrophic mutations in this organism. Genetic mapping allowed us to identify two new genes, pur-8001 and met-8003. The Tn501-induced auxotrophic mutations were distributed non-randomly among auxotrophic genes, and the reversion of the mutations by precise excision of the Tn501 insertion occurred very rarely.

Bacteriophages↗

Purification of a eukaryotic site-specific endonuclease, Endo.Sce I, from Saccharomyces cerevisiae and effectors on its specificity and activity.

A site-specific endonuclease (Endo.Sce I) which caused double-strand scission of DNA was highly purified from a eukaryote, Saccharomyces cerevisiae IAM4274. The molecular weight of the active form of Endo.Sce I was estimated to be 120,000 and 110,000 by sedimentation analysis on a glycerol density gradient and gel filtration on Ultrogel AcA34, respectively. Analysis of the fractions from the last column chromatography by polyacrylamide gel-electrophoresis in the presence of sodium dodecyl sulfate and by an assay of the endonucleolytic activities suggested that Endo.Sce I consists of two non-identical subunits with molecular weights of 75,000 and 50,000. Unlike restriction endonucleases, Endo.Sce I was active on chromosomal DNA of the cells which produced Endo.Sce I. Single-stranded DNA was not cleaved by Endo.Sce I, but inhibited the endonucleolytic activity of the enzyme on double-stranded DNA. The endonucleolytic activity of Endo.Sce I required the magnesium ions (Mg2+) as a sole cofactor; Mg2+ could not be replaced by Ca2+ or Zn2+. When Mg2+ was replaced by manganese ions (Mn2+), extensively purified Endo.Sce I cleaved double-stranded DNA at many other sites in addition to the sites at which DNA was cleaved in the presence of Mg2+. Experiments indicated that this is not the activation of contaminating endonuclease in the preparation of Endo.Sce I, but the result of relaxation in the site-specificity of cleavage.

Base Sequence↗

Genetic analysis of three additional fla genes in Salmonella typhimurium.

In Salmonella typhimurium, 27 fla genes responsible for formation of flagella have been identified and assigned to three regions on the genetic map, termed fla regions I to III. By genetic analysis of 1984 non-flagellate mutants obtained from a phase-1 stable strain of S. typhimurium, SJW1103, three additional fla genes were identified; one, termed flaW, was assigned to fla region I and the other two, termed flaV and flaX, to fla region III. By intergeneric complementation tests, the flaW, flaV and flaX genes were shown to be functionally homologous with flaS, flbC and flaP of Escherichia coli, respectively. Electron microscopy showed that flaW and flaV mutants carried hook-basal body structures.

Chromosome Mapping↗

Genetic analysis of H2, the structural gene for phase-2 flagellin in Salmonella.

Non-flagellate H2 mutants were isolated from a phase-2 stable strain, SJW806 H1-gt- H2-enxon vh2-, a derivative of Salmonella typhimurium. By transductional crosses a deletion map and a recombination map of the H2 gene were made. There are three regions especially rich in nonflagellate mutational sites. By the use of the deletion map, mutational sites of 21 flagellar shape mutants were also determined. Most of them were located at two regions which coincide with two of the three regions rich in non-flagellate mutational sites. A gene, vh2, is closely linked to the promoter side of the H2 gene. Three-factor transductional crosses showed that the vh2 gene was on the left of the H2 gene in the present map. The H2 gene forms part of an operon with the distal gene rh1 which specifies the H1 repressor. Thus, a polarity effect of the H2 mutations on the expression of the rh1 gene was examined by observing whether a wild-type H1 allele introduced into the H2 mutants was expressed or not. Many of the H2 mutations were polar, and most of the strongly polar mutations were located in the left (promoter-proximal) half of the H2 gene, while most of the mutations in the right half of the gene were weakly polar or non-polar.

Bacterial Proteins↗

Hook-associated proteins essential for flagellar filament formation in Salmonella typhimurium.

The hooks of the flagella of Salmonella typhimurium were purified by a newly developed method, using a flaL mutant without a filament, and the hook components were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. As a result, we detected three protein species in addition to hook protein. We call these three proteins hook-associated proteins (HAPs). Their molecular weights were 59,000 for HAP1, 53,000 for HAP2, and 31,000 for HAP3. The HAP1/hook protein/HAP3/HAP2 molar ratio, calculated from their relative amounts and their molecular weights, was 1:10:1.1:0.53. The compositions of HAPs were analyzed in the hooks from the other filamentless mutants which were defective in H1 H2, flaV, flaU, or flaW. Hooks from the H1 H2 mutant had the same HAP composition as hooks from the flaL mutant. Hooks from the flaV mutants contained HAP1 and HAP3. Hooks from the flaU mutants contained HAP1. Hooks from the flaW mutants contained a very small amount of HAP3. From these results, the process of hook morphogenesis and the genes responsible for each step were postulated. Electron micrographs of hooks from the filamentless mutants showed that hooks which contained all three HAPs had a sharp clawlike tip, whereas hooks lacking any HAP had a flat tip. Electron micrographs of hooks treated with antibody against the hook protein showed that each claw-shaped end was not covered with antibody. These results strongly suggest that all three HAPs or at least some of them are located at the claw-shaped end and play an essential role in filament formation.

Bacterial Proteins↗

Molecular cloning and characterization of genes required for ribose transport and utilization in Escherichia coli K-12.

We isolated spontaneous and transposon insertion mutants of Escherichia coli K-12 that were specifically defective in utilization or in high-affinity transport of D-ribose (or in both). Cotransduction studies located all of the mutations near ilv, at the same position as previously identified mutations causing defects in ribokinase ( rbsK ) or ribose transport ( rbsP ). Plasmids that complemented the rbs mutations were isolated from the collection of ColE1 hybrid plasmids constructed by Clarke and Carbon. Analysis of those plasmids as well as of fragments cloned into pBR322 and pACYC184 allowed definition of the rbs region. Products of rbs genes were identified by examination of the proteins produced in minicells containing various rbs plasmids. We identified four rbs genes: rbsB , which codes for the 29-kilodalton ribose-binding protein; rbsK , which codes for the 34-kilodalton ribokinase ; rbsA , which codes for a 50-kilodalton protein required for high-affinity transport; and rbsC , which codes for a 27-kilodalton protein likely to be a transport system component. Our studies showed that these genes are transcribed from a common promoter in the order rbsA rbsC rbsB rbsK . It appears that the high-affinity transport system for ribose consists of the three components, ribose-binding protein, the 50-kilodalton RbsA protein, and the 27-kilodalton RbsC protein, although a fourth, unidentified component could exist. Mutants defective in this transport system, but normal for ribokinase , are able to grow normally on high concentrations of the sugar, indicating that there is at least a second, low-affinity transport system for ribose in E. coli K-12.

Bacterial Proteins↗

Transformation of L cells with virus thymidine kinase genes introduced by red cell-mediated microinjection.

Fusion of red cell ghosts containing foreign materials with cells results in the introduction of the materials into the cells (red cell-mediated microinjection). Until now, 'two-step dialysis' has mainly been used for trapping proteins in the ghosts. Large-sized materials such as DNA, however, are rarely trapped in the ghosts, since the holes in the red cell membrane caused by osmotic shock are too small for such materials to pass through. In this study, we improved the trapping technique. Some of the Hind III fragments of lambda phage DNA as well as proteins could be trapped in the ghosts when the mixture of these materials and red cells were frozen at -80 degrees C for a short period followed by quick thawing. Red cell-mediated microinjection using ghosts containing plasmid pBR322 linked with a Herpes simplex viral thymidine kinase (tk) gene brought about transformation of tk-defective L cells, the efficiency of transformation was 1 out of 20 000-60 000 cells fused with the ghosts.

Animals↗