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Biomedical subjects

Y Sakanoue

Publications and source records attributed to Y Sakanoue.

68 records · Page 4Linked to original sources

Conversion of malfunctioning J pouch to Kock's pouch. Case report.

A patient with ulcerative colitis previously treated with restorative proctocolectomy and ileal J-pouch anal anastomosis had recurrent pelvic abscess and poor functional results for 3 years postoperatively. To improve the quality of life, the J pouch was converted to a Kock's pouch. The successful result implies that Kock's continent ileostomy is a good replacement for failed pelvic pouch.

Abscess↗

Insulin-stimulated serine kinase in Xenopus oocyte plasma membrane.

Insulin-stimulated protein kinase activities detected in Xenopus oocyte membrane were examined. The plasma membrane proteins solubilized in a buffer containing Triton X-100 were immunoprecipitated with anti-phosphotyrosine antibodies and adsorbed materials were eluted with a buffer containing p-nitrophenyl phosphate. The eluate contained protein serine kinase activity toward H1 histone which was increased 2-3 fold by insulin. Protein tyrosine kinase activity was also exhibited in Xenopus oocyte membrane and the close parallel to serine kinase activity was observed in response to insulin. These results suggest that insulin-stimulated serine kinase is activated through the phosphorylation by protein tyrosine kinase.

Animals↗

Ionic strength-dependent proteolytic activation of protein kinase C by trypsin-like protease.

Protein kinase C, reversibly bound to rat liver plasma membrane through Ca2+, was activated by endogenous trypsin-like protease in an ionic strength-dependent manner. In an attempt to understand the reaction mechanism, the EGTA-extracted protein kinase C and the trypsin-like protease (Tanaka, K. et al. (1986) J. Biol. Chem. 261, 2610-2615) were separately purified from plasma membrane. In the reaction system using these purified enzymes, increasing the ionic strength with NaCl (140-210 mM) effectively enhanced the proteolytic activation of the protein kinase C in the presence of Ca2+ and phospholipid. These results suggest that ionic strength is an important factor for the proteolytic activation of membrane-bound rat liver protein kinase C.

Animals↗

Comparison of phosphorylation sites in protamines between protein kinase C and cAMP-dependent protein kinase.

Phosphorylation sites of protamines by protein kinase C and cAMP-dependent protein kinase (protein kinase A) were studied. Using clupeine Y1 as a substrate, protein kinase C phosphorylates both Ser and Thr residues, whereas protein kinase A phosphorylates only Ser residue(s). Protein kinase C phosphorylates all Ser and Thr residues of clupeine Y2 and Z, however protein kinase A phosphorylates mainly Ser9 and slightly Thr5 in clupeine Y2 and Ser6 and Ser10 in clupeine Z. These results suggest that protein kinase C recognizes more sites than those of protein kinase A and may participate in protamine phosphorylation in vivo.

Amino Acid Sequence↗

Comparative studies on phosphorylation of synthetic peptide analogue of ribosomal protein S6 and 40-S ribosomal subunits between Ca2+/phospholipid-dependent protein kinase and its protease-activated form.

Ca2+/phospholipid-dependent protein kinase (protein kinase C) and trypsin-activated protein kinase C (protein kinase M) phosphorylated the synthetic peptide R1-A13 (Arg-Arg-Leu-Ser-Ser-Leu-Arg-Ala-Ser-Thr-Ser-Lys-Ala) which contains both cAMP- and insulin-regulated phosphorylation sites in rat liver ribosomal protein S6 [Wettenhall, R. E. H. & Morgan, F. J. (1984) J. Biol. Chem. 259, 2084-2091]. Both enzymes showed essentially the same kinetic properties; V and apparent Km were determined to be 0.16 mumol min-1 mg-1 and 30 microM, respectively. At first, tryptic phosphopeptides were prepared at the early stage of phosphorylation and purified by high-performance liquid chromatography (HPLC). Through these analyses, four radioactive peptides were isolated. When protein kinase C was employed, phosphorylation was observed on all four peptides in a Ca2+/phospholipid-dependent manner. Irrespective of the protein kinase employed, phosphate incorporation into these peptides increased linearly with time; the peptide concentration did not affect the ratio of phosphate distribution into these four peptides. Analysis of amino acid composition and phosphoamino acid of radioactive peptides obtained after extensive phosphorylation showed that phosphates were incorporated into Ser-4, Ser-5, Ser-9 and Ser-11. The latter three serine residues were major phosphorylated sites. When rat liver 40-S ribosomal subunits were employed as substrate for protein kinases C and M, a radioactive protein with Mr,app = 31,000, which corresponded to S6 protein, was detected on an autoradiogram of a sodium dodecyl sulfate/polyacrylamide slab gel. The rate of phosphorylation with protein kinase M was twice as fast as that with protein kinase C. The elution profile of radioactive tryptic peptides in HPLC suggest that phosphorylation occurred on the sites in S6 protein corresponding to Ser-5, Ser-9 and Ser-11 as major sites and Ser-4 as the minor one. These results indicate that protein kinase C has an ability to recognize at least four sites derived from hormone-dependent phosphorylation sites in ribosomal protein S6 irrespective of the mode of activation of this enzyme.

Amino Acid Sequence↗

An activated S6 kinase in regenerating rat liver.

S6 kinase activity was increased in the regenerating liver 5 h after partial hepatectomy compared with sham-operated liver. The protein kinase activity was eluted from DE-52 column at approximately 250 mM NaCl and was not affected by known regulators of protein kinases. The S6 kinase was further purified by chromatography on peptide R1A13-Sepharose 4B and Sephadex G-150. The molecular weight of the enzyme was estimated to be 4.5 X 10(4) by gel filtration. The enzyme catalyzes the phosphorylation of whole histone, mainly H2B histone, at 75 mM Mg2+. These properties are similar to those of a proteolytically modified Ca2+/phospholipid-independent form of protein kinase C.

Amino Acid Sequence↗

Non-requirement of calcium on protamine phosphorylation by calcium-activated, phospholipid-dependent protein kinase.

Phosphorylation of clupeine sulfate by purified rat brain calcium-activated, phospholipid-dependent protein kinase (protein kinase C) was studied. In the absence of Ca2+, phosphatidylserine and diolein markedly stimulated its phosphorylation. However Ca2+ did not stimulate but inhibit this phosphorylation about 30% in the presence of phospholipids. Random polymer (Arg, Ser) 3:1 and (Lys, Ser) 3:1 could be phosphorylated by protein kinase C. In the presence of phospholipids Ca2+ is not needed for the phosphorylation of polymer (Arg, Ser) 3:1, while Ca2+ is necessary for polymer (Lys, Ser) 3:1. Non-requirement of Ca2+ on clupeine phosphorylation by protein kinase C is briefly discussed.

Calcium↗

Effect of ionic strength on production of cAMP- and Ca2+-independent protein kinase from rat liver plasma membrane.

Production of cAMP- and Ca2+-independent protein kinase was stimulated when rat liver plasma membrane was incubated with increasing concentrations of NaCl. This protein kinase release was diminished by addition of protease inhibitor. The molecular mass of this enzyme was approx. 50 kDa and a high concentration of Mg2+ was required for whole histone phosphorylation. These properties are similar to those of the protease-activated form of protein kinase C. The NaCl effect could be replaced by other salts such as LiCl and NaHCO3. These results suggest that membrane-bound protein kinase C is activated by limited proteolysis corresponding to an increase in ionic strength.

Animals↗

Intraoperative colonoscopy.

From 1987 to 1991, we performed intraoperative colonoscopy on 66 patients as a result of the following indications: the air leakage test (53%), detection of a previous malignant polypectomy site (20%), inability to complete preoperative colonoscopy (17%), detection of the source of intestinal bleeding (4%), and detection of impalpable colonic lesions (4%). Intraoperative colonoscopy was successful in 61/66 patients and provided information that altered the planned operation in 10 of 61 completely examined patients. The air leakage test proved useful in detecting subclinical anastomotic leaks. The indications for intraoperative colonoscopy have been expanded, and this procedure is often useful when one is attempting to decide on the appropriate surgical strategy for patients.

Anastomosis, Surgical↗

Dobutamine-stress electrocardiographically gated positron emission tomography for detection of viable but dysfunctional myocardium.

PURPOSE: This study was performed to determine whether low-dose dobutamine stress electrocardiography (ECG)-gated fluorine-18 fluorodeoxyglucose (FDG)-positron emission tomography (PET) can assess wall motion and identify myocardium without contractile reserve despite preserved FDG uptake. METHODS: Fifty-three patients with myocardial infarction and normal sinus rhythm underwent ECG-gated FDG-PET and transthoracic echocardiography. Wall motion of 10 segments of the left ventricle was graded as normal, hypokinetic, or akinetic/dyskinetic. RESULTS: In 365 (76%) of 480 segments, assessment of wall motion was concordant between the 2 modalities. In 30 patients dobutamine-stress ECG-gated FDG-PET was performed. In 13 (50%) of 26 dysfunctional segments with normal FDG uptake, 16 (36%) of 44 dysfunctional segments with mildly reduced FDG uptake and 12 (25%) of 48 dysfunctional segments with moderately reduced FDG uptake, wall motion was improved by dobutamine infusion. CONCLUSION: Assessment of left ventricular wall motion with ECG-gated FDG-PET is feasible, and dobutamine stress ECG-gated FDG-PET can simultaneously identify metabolic viability and contractile reserve.

Adult↗

Dobutamine stress electrocardiography-gated Tc-99m tetrofosmin SPECT for detection of viable but dysfunctional myocardium.

BACKGROUND: Technetium-99m-labeled myocardial perfusion tracers allow simultaneous assessment of myocardial perfusion and left ventricular function by electrocardiography-gated scan. This study was performed to determine whether dobutamine stress electrocardiography-gated tetrofosmin single photon emission computed tomography (SPECT) can identify viable (as defined by positron emission tomography [PET]) but dysfunctional myocardium with contractile reserve. METHODS AND RESULTS: Thirty-five patients with myocardial infarction underwent resting electrocardiography-gated SPECT and fluorodeoxyglucose (FDG) PET. The relative uptakes of tetrofosmin (%tetrofosmin) and FDG (%FDG) were calculated. Wall motion in 9 left ventricular segments was assessed at rest and during dobutamine stress on a 3-dimensional cine-mode display created with automatic left ventricular function analysis software. A total of 129 dysfunctional segments were analyzed. Forty-five (48.9%) of 92 segments with %tetrofosmin of 50% or greater and only 4 (10.8%) of 37 segments with %tetrofosmin less than 50% had contractile reserves (P <.0001). The sensitivity, specificity, and predictive accuracy of %tetrofosmin of 50% or greater for detecting %FDG of 50% or greater were 85.7%, 74%, and 82.9%, respectively. The incidence of the presence of contractile reserve rose with increasing magnitude of %FDG. The sensitivity, specificity, and predictive accuracy of the presence of contractile reserve for detecting %FDG of 50% or greater were 43.9%, 80.6%, and 52.7%, respectively. CONCLUSIONS: Dobutamine stress electrocardiography-gated SPECT can identify viable (as defined by PET) but dysfunctional myocardium with contractile reserve.

Adult↗

The efficacy of whole gut irrigation with polyethylene glycol electrolyte solution in elective colorectal surgery for cancer.

Sixty-four patients undergoing elective operations for cancer of the colon and rectum were given mechanical bowel preparation in the form of whole gut irrigation with polyethylene glycol electrolyte solution. When their colonic cleansing scores were compared, it was found that the preparation was significantly less effective in the left colon (n = 49) than in the right (n = 15), the mean (SEM) scores being 3.8 (0.56) and 2.65 (1.01), respectively, p less than 0.001. We then compared the scores of 26 patients with stenosing lesions with those of 38 that were not: in the right colon the score for stenosing tumours was 3.9 (0.14) compared with 3.8 (0.25) for non-stenosing tumours. In the left colon, however, they were 2.3 (0.23) and 2.9 (0.18), respectively (p less than 0.01). We conclude that stenosis of the colon caused by malignant lesions reduces the efficacy of mechanical bowel preparation with polyethylene glycol electrolyte solution in the left colon but not in the right colon.

Colon↗