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Biomedical subjects

H Sakai

Publications and source records attributed to H Sakai.

At least 1,009 records · Page 56Linked to original sources
PubMed1989

[Study on metabolism of pyridonecarboxylic acid II. Determination of metabolites of (+-)-7-(3-amino-1-pyrrolidinyl)-6-fluoro-1-(2,4-difluorophenyl)- 1,4-dihydro-4-oxo-1,8-naphthyridine-3-carboxylic acid p-toluenesulfonate hydrate (T-3262) in blood, urine, bile, and feces].

The fate of (+-)-7-(3-amino-1-pyrrolidinyl)-6-fluoro-1-(2,4-difluorophenyl-1,4- dihyro-4-oxo-1,8-naphthyridine-3-carboxylic acid p-toluenesulfonate hydrate (T-3262) was studied using T-3262 and 14C-T-3262 in various animals. 1. Metabolites in serum and urine were assayed for mouse, rat, rabbit, dog and monkey following oral administration of T-3262. In serum, besides unchanged T-3262 base, T-3262A (N-acetylated) was detected in rat, rabbit and monkey; T-3262B (deamino-hydroxylated) was detected in monkey. In urine, unchanged T-3262 base was excreted mainly. But a few of metabolites (T-3262A, T-3262B, T-3262 glucuronide, T-3262A glucuronide, T-3262B glucuronide, and unknown compound M-1) were detected, and species difference existed in types of metabolites. 2. Metabolites in bile and feces were assayed for mouse and rat following oral administration of T-3262 and 14C-T-3262. Metabolites in bile were similar to the urine, but the volume of T-3262A and T-3262A glucuronide was larger than in urine. In feces, the excreted compounds mainly consisted of unchanged T-3262 base. 3. p-Toluenesulfonic acid, which is the counter acid for T-3262 base, was absorbed following the oral administration of T-3262, and excreted in urine in the unchanged form.

4-Quinolones↗
PubMed1989

Glomerular T cell and monocyte populations in patients with IgA nephropathy.

Renal tissues from 19 patients with the minimal or slight stage of IgA nephropathy were examined for evidence of glomerular T cell or monocyte infiltration using monoclonal antibodies to identify T cells (OKT3, OKT11), T cell subsets (OKT4, OKT8), and monocytes and null cells (OKM1) by indirect immunofluorescence. Renal tissues from 12 patients at the same stage of mesangial proliferative glomerulonephritis without IgA deposition were also investigated by these procedures. Light microscopic examination of the same renal tissues was also performed. Reactive glomerular mononuclear cells were found to be numerous in patients with IgA nephropathy. The most prominent type of glomerular cells was OKT8 positive. T cell subsets and OKM1 positive cells in glomeruli from patients with IgA nephropathy were significantly increased as compared to those from patients with mesangial proliferative glomerulonephritis. Focal segmental proliferation of mesangial cells was observed in glomeruli which showed an accumulation of OKT8 positive cells. It is concluded that the immuno-regulatory mechanism involving T cells and/or monocytes might be one of the exacerbating factors in patients with IgA nephropathy.

Glomerulonephritis, IGA↗
PubMed1989

[A portable manual sternal compressor for cardiac massage of a patient under transportation--1. Evolution after trials and errors].

Difficulties of external cardiac massage during transportation of a patient on a litter or stretcher prompted us to develop a new portable manual sternal compressor. A seemingly good idea of sternal compression with an inflatable balloon placed between the sternum and a chest band engaged us in a long ordeal of trial and error until we retreated from this preconception. A practical sternal compressor has finally been created by applying two hydraulic pistons, one as a sternal compressor and the other as its push-pull energizer incorporated into a manual lever system. In this paper, one of the serial reports on this new device, all our errors are described lest some one in the future meet the same problems. Details of this new apparatus and the results of our experimental study and clinical application will soon follow this paper.

Equipment Design↗
PubMed1989

[A study on the distribution of T-3262 (tosufloxacin tosilate) in salivary glands of rats using microautoradiography].

The distribution of T-3262 (tosufloxacin tosilate) in salivary glands of rats was investigated with frozen-microautoradiography. One and 4 hours after oral administration of 14C-T-3262 at 100 mg/kg to rats submandibular glands, parotid glands and sublingual glands were removed, and a microautoradiogram of each was made. In the submandibular gland and the parotid gland 14C-T-3262 was distributed at high levels throughout the glands taken at 1 and 4 hours after administration, but lower levels than the other glands were found in the sublingual gland at 1 hour. The results of this study suggested that T-3262 penetrates effectively into the saliva, because 14C-T-3262 is distributed well into glandular acinus, striated duct and excretory duct. The microautoradiography was a useful and reliable method for investigating the distribution of antimicrobial agents in salivary glands.

4-Quinolones↗
PubMed1989

[Surgical indication to deep-seated arteriovenous malformation].

The natural history of cerebral arteriovenous malformation (AVN) is still a subject of dispute. In the case of deep seated AVM ruptures, the problem is more serious because the hemorrhages often give rise to severe disability or death, depending on their location. Total extirpation of the nidus is fundamentally the best choice for treatment of AVM. It is never acceptable, however, to allow a persistent post operative deficit. Forty-seven cases of deep AVM were encountered in the Tokyo Jikei University Hospital. Total extirpation of the nidus was successful in 32 cases, while no surgery was undertaken in 15. Up to the present time we have taken three risk factors into condition in judging the indications for deep seated AVM surgery: the size of the nidus, the position of the nidus, and the pattern of demarcation around nidus, as shown on CT scan. When the nidus is bordered by a hemorrhage, cyst, or ventricular wall, dissection of the nidus is facilitated and injury to adjacent brain tissue is minimized, and this is why we included profound demarcation around the nidus as the third factor. We then determined grade of risk with respect to each factor, and counted a risk score in each case. The long-term result was judged by the response to questionnaires. Of the 15 patients who did not undergo surgery, four died on account of rebleeding. The longer the patient having deep AVM survived, the more likely the occurrence of fatal rebleeding. Of the 32 patients operated on, two died immediately after surgery, while 23 survived in an improved state or without any aggravated neurological deficit.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗
PubMed1988

Role of the potential secondary structures in phage G4 origin of complementary DNA strand synthesis.

Phage G4 origin of complementary DNA strand synthesis (oric) consists of three stable stem-loop structures (I, II, and III). Mutant oric sequences with alterations in the structure of stem-loop II, stem-loop III, and the stem-loop II-III spacer region have been constructed and cloned into the filamentous phage vectors to assay their functional activity. Changes in the lowermost GC base pair in the stem of stem-loop III, in the 9-bp spacer region between the stems of stem-loops II and III, and in the loop of stem-loop II, impair or abolish in vivo oric function. The results suggest that recognition sequences for dnaG primase must be present in the loop of stem-loop II, and in the spacer region between the stems of stem-loops II and III.

Bacteriophages↗
PubMed1988

Purification and characterization of the 90-kDa heat-shock protein from mammalian tissues.

The 90-kDa heat-shock protein (HSP90) has been purified from mammalian tissues, mouse liver and porcine brain, with a good yield by a new method involving hydrophobic chromatography. Mouse liver HSP90 and porcine brain HSP90 were compared with mouse lymphoma HSP90 which was purified from T lymphoma cell line, L5178Y, by a modification of the previously reported method. These three HSP90s were indistinguishable from one another in amino acid composition, one-dimensional peptide mapping, elution pattern of proteolytic fragments (trypsin- or V8-protease-cleaved) in reverse-phase high-performance liquid chromatography, reactivity with the antibody against mouse T lymphoma HSP90 and the ability to bind to F-actin. The amino acid sequences of three portions (total 47 amino acid residues) of lymphoma HSP90 were determined and they were homologous to those of the corresponding portions of Drosophila HSP83A and yeast HSP90. These results suggest that HSP90 is a highly conserved protein during evolution.

Amino Acid Sequence↗
PubMed1988

Microtubule-assembly inhibitor protein. Its distribution, localization and physicochemical properties.

Microtubule-assembly inhibitor protein (MIP) is an acidic protein with Mr 33,000 which inhibits microtubule assembly in vitro [Kotani, S., Murofushi, H., Nishida, E. & Sakai, H. (1984) J. Biochem. (Tokyo) 96, 959-969]. Anti-MIP antibody was affinity-purified from rabbit anti-MIP sera raised against chemically modified MIP. MIP was localized in the nucleus in interphase culture cells as revealed by immunofluorescent light microscopy. Immunoblotting experiments showed that MIP exists in a variety of mammalian cells and tissues. Kidney appeared to be a better source of MIP than brain, the original source. Kidney MIP was isolated by the same procedure as for brain MIP and proved to be indistinguishable from brain MIP in the inhibitory activity of microtubule assembly, molecular mass, immunoreactivity, and one-dimensional peptide mapping. Physico-chemical characteristics of MIP were studied using the kidney protein. It contained 20% aspartic acid and 25% glutamic acid, accounting for its acidic nature. Hydrodynamically, MIP was a monomer with S20,w = 1.9 S and Mr = 30,000. The frictional ratio, f/fo = 1.7, indicated that MIP is not a globular molecule but has either an elongated or an expanded structure. Circular dichroic results showed a low content of alpha-helix or beta-sheet structure for MIP. Proton nuclear magnetic resonance analysis provided evidence that MIP consists mainly of very flexible structures (random-coil-like structures), but still contains a hydrophobic core structure below 60 degrees C.

Amino Acids↗
PubMed1988

Relationships between axonal diameter, soma size, and axonal conduction velocity of HRP-filled, pyramidal tract cells of awake cats.

Relationships between axonal diameter, soma size, and axonal conduction velocity were examined in intracellularly recorded pyramidal tract (PT) cells of conscious cats using pressure injection of horseradish peroxidase. Positive linear correlations were found between axonal conduction velocities and axonal diameters as well as between axonal conduction velocities and soma sizes. All PT cells had somata located in layer V. Slow PT cells had high densities of dendritic spines in layer III; however, so did some fast PT cells, making this morphologic feature unacceptable for distinguishing between slow and fast conducting PT neurons.

Animals↗
PubMed1988

Two single-strand DNA initiation signals located in the oriV region of plasmid RSF1010.

Two single-strand initiation signals (ssi) are found in the oriV region of broad-host-range plasmid RSF1010, using a plaque assay system with a mutant M13 phage which lacks the greater part of the complementary DNA strand origin (oric). These two signals, designated ssiA and ssiB, have RSF1010-specific properties, because they require one or more RSF1010-specific factors provided in trans. The functional activity of ssiA is higher than that of ssiB. The two signals are located on separate DNA strands, so that the DNA chain elongations initiated from them in the opposite directions may pass each other. It is conceivable that these signals, ssiA and ssiB, direct DNA priming functions at the initiation stage in vegetative DNA replication of RSF1010.

Base Composition↗
PubMed1988

Purification and characterization of kinesin from bovine adrenal medulla.

Kinesin was purified from bovine adrenal medulla. The sedimentation coefficient was 8.8 S. Sedimentation equilibrium ultracentrifugation studies showed the molecular weight of kinesin to be 300,000. The calculated axial ratio was 1:16. The Stokes radius was estimated to be 8.9 nm by gel filtration. Circular dichroism showed the alpha-helix content to be about 50%. Purified kinesin preparation contained a major polypeptide with a molecular weight of 120,000 and minor ones with molecular weights of 71,000, 68,000, and 65,000. Bovine adrenal kinesin had an ATPase activity which was stimulated severalfold by microtubules to a specific activity of about 0.1 mumol/min.mg. Kinesin molecules adsorbed to a glass slide promoted the movement of microtubules on the glass surface at a rate of about 0.5 micron/s. Immunostaining of EBTr (bovine embryonic trachea fibroblast) cells and bovine adrenal chromaffin cells in interphase with an affinity-purified antibody against the major polypeptide of kinesin showed that some kinesin was located on microtubules and the rest distributed throughout the cytoplasm in a diffuse manner. EBTr cells in mitotic phase gave a staining pattern showing that kinesin was present throughout the cytoplasm with higher concentration in the region of mitotic apparatus.

Adenosine Triphosphatases↗
PubMed1988

Colchicine acts as a progression factor to initiate DNA synthesis in quiescent Balb/c 3T3 cells.

In quiescent Balb/c 3T3 cells, competence factors such as 12-O-tetradecanoylphorbol-13-acetate (TPA) and platelet-derived growth factor (PDGF) synergize with progression factors such as insulin to initiate DNA synthesis. In this study, we found that colchicine, a microtubule-disrupting agent, acted synergistically with TPA, but not with insulin, to induce the maximal stimulation of DNA synthesis. Colchicine also synergized with PDGF in the presence of epidermal growth factor to elicit nearly the optimal induction of DNA synthesis. Moreover, it acted synergistically with fibroblast growth factor, another competence factor. These results suggest that colchicine acts as a progression factor like insulin in quiescent Balb/c 3T3 cells.

Animals↗
PubMed1988

Cloning and characterization of porcine brain cofilin cDNA. Cofilin contains the nuclear transport signal sequence.

Cofilin is a widely distributed, pH-sensitive, actin-modulating protein with an apparent molecular mass of 21 kDa, which forms intranuclear and/or cytoplasmic actin/cofilin rods in cultured fibroblastic cells under specific conditions. In this study, a cDNA library from porcine brain mRNA was constructed, and full-length brain cofilin cDNA clones were isolated by screening with oligonucleotide probes. The deduced amino acid sequence of cofilin is 166 residues long and contains a sequence of Lys-Lys-Arg-Lys-Lys which is very similar to the nuclear transport signal sequence (Pro-Lys-Lys-Lys-Arg-Lys-Val) of SV40 large T antigen. The sequence may act as a signal capable of inducing nuclear accumulation of cofilin in cells exposed to heat shock or dimethyl sulfoxide. The cofilin sequence contains a hexapeptide (Asp-Ala-Ile-Lys-Lys-Lys) identical to the amino-terminal sequence (residues 2-7) of muscle and nonmuscle tropomyosin. Cofilin also has in the carboxyl-terminal portion a region homologous to the sequence shared by gelsolin, fragmin, and Acanthamoeba profilin. Furthermore, the overall amino acid sequence of cofilin shows weak homology with the rod portion of myosin and suggests a high alpha-helical content.

Actin Depolymerizing Factors↗
PubMed1988

Microtubule-binding domain of tau proteins.

Limited chymotryptic digestion of whole tau proteins produced a fragment of Mr 14,000 (CT14), which was able to bind to microtubules reconstituted from tubulin alone in the presence of taxol. This fragment was also found to persist in microtubules when microtubules consisting of tau proteins and tubulin were digested by chymotrypsin. Analysis of amino acid composition revealed that CT14 was rich in lysine and proline residues, suggesting unique structure of microtubule-binding domain of tau proteins. Amino-terminal sequence of CT14 was determined to be Ser-Ser-Pro-Gly-Ser-Pro-Gly-Thr-Pro-Gly-Ser-Arg-Ser-Arg-X-Pro-Ser-Leu-Pr o. No heterogeneity was detected in this amino-terminal sequence of 19 residues. Five species of polypeptides consisting of tau proteins were separated from each other by gel electrophoresis and subjected to chymotryptic digestion. CT14 was produced from each of the tau polypeptides by chymotryptic digestion, indicating that all tau polypeptides have a common microtubule-binding domain.

Amino Acids↗
PubMed1988

Protein-kinase-C-catalyzed phosphorylation of the microtubule-binding domain of microtubule-associated protein 2 inhibits its ability to induce tubulin polymerization.

It has previously been demonstrated that microtubule-associated protein 2 (MAP2) is a good substrate for the purified protein kinase C [Tsuyama, S., Bramblett, G. T., Huang, K.-P. & Flavin, M. (1986) J. Biol. Chem. 261, 4110-4116; Akiyama, T., Nishida, E., Ishida, J., Saji, N., Ogawara, H., Hoshi, M., Miyata, Y. & Sakai, H. (1986) J. Biol. Chem. 261, 15648-15651]. We have shown here that phosphorylation of MAP2, catalyzed by protein kinase C, reduces the ability to induce tubulin polymerization. MAP2 is divided into two domains by digestion with alpha-chymotrypsin; the microtubule-binding and the non-binding (projection) domains. The limited chymotryptic digestion following phosphorylation of MAP2 by protein kinase C has shown that both the domains of MAP2 were phosphorylated by protein kinase C, 50-60% of the incorporated phosphates being detected in the microtubule-binding domain. Polypeptide fragments, containing the microtubule-binding domain of MAP2, were purified by DEAE-cellulose column chromatography after chymotryptic digestion of MAP2. The purified microtubule-binding fragments were competent to polymerize tubulin, and served as good substrates for protein kinase C. The phosphorylation of the microtubule-binding fragments by protein kinase C reduced their ability to induce tubulin polymerization. These results suggest that the ability of MAP2 to induce tubulin polymerization is inhibited by phosphorylation of the microtubule-binding domain catalyzed by protein kinase C.

Actins↗
PubMed1988

Plasmid pACYC184 contains an ssi signal for initiation of single-strand phage DNA replication.

Using the plaque assay system for screening the single-strand (ss) initiation determinant (ssi) sequences, we have found that 119-bp region in pACYC184, a derivative of the plasmid P15A of Escherichia coli, can direct such ss DNA initiation. This region is located downstream from the P15A origin of replication and conserves consensus sequences of the ssi signals found in the other plasmids. Signals for ss DNA initiation are defined as nucleotide sequences present on ss DNA templates and required for priming DNA synthesis. The direction of chain elongation in DNA synthesis is opposite to that of the leading strand. In this region, we found a potential stem-and-loop structure. The 119-bp DNA segment of plasmid pACYC184 cloned in f1R199 filamentous phage could direct rifampicin-resistant conversion of the ss DNA to the double-stranded replicative form.

Base Sequence↗