[Site-specific microbiological tests and the diagnostic significance-- bacteremia, sepsis and bacterial endocarditis].
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Biomedical subjects
Publications and source records attributed to M Yamane.
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Median sternotomy is the standard incision for surgery of the heart and great vessels. But patients who undergo median sternotomy continue to suffer morbidity and death from sternal wound complications. Recently we have experienced 4 cases of patients with postoperative sternal dehiscence following median sternotomy. We have used a technique of sternal approximation that is a modification of the technique described by Robicsek and associates. We believe this technique should not be used routinely but seems to be useful in the stabilization of complicated sternal dehiscence especially in the presence of multiple cutting through the sternum. The described technique is easy to perform, and its advantage is that it provides good stability in perioperative respiratory management of patient with sternal dehiscence following median sternotomy.
A homogenate of rat brain, rat liver or human colonic well differentiated adenocarcinoma was prepared in 250 mM sucrose isoosmolaric buffer (pH 7.6) and fractionated by differential centrifugation at 10(3), 10(4) and 10(5) g. Each precipitate or supernatant was incubated with NADPH and docosahexaenoic acid or arachidonic acid as a substrate for 30 min at 37 degrees c under aerobic conditions. omega-Hydroxydocosahexaenoic acid or omega-hydroxyeicosatetraenoic acid from an incubation mixture was detected by reversed-phase high-performance liquid chromatography-thermospray mass spectrometry with selected-ion monitoring. omega-Hydroxy polyunsaturated fatty acids were characterized by high intensity of the molecular ion (MH+) although common hydroxy polyunsaturated fatty acids were characterized by high intensity of the MH+ -H2O ion. For the rat brain, omega-hydroxylation activity (the amount of omega-hydroxy product produced in 30 min) was concentrated to a 10(3) g precipitate although the specific activity (the activity per 1 mg of protein) in the 10(3) g precipitate did not indicate superiority over other fractions. However, the specific activity of the rat brain increased on addition of a 10(4) or 10(5) g precipitate. For the rat liver, although omega-hydroxylation activity was concentrated to a 10(3) g precipitate, the specific activity was concentrated to a 10(5) g precipitate and the subcellular localization differed from that of rat brain. In the human colonic well differentiated adenocarcinoma, although omega-hydroxylation activity was relatively high in the 10(3) g supernatant, the specific activity was relatively high in the 10(3) g precipitates. These results suggest that there is a difference regarding subcellular localization of the omega-hydroxylation activity depending on the species of the organs.
We have developed a method for the analysis of 5,6-dihydroxyeicosatrienoate-1,5-lactone (5,6-DiHETriE-delta-lactone) in tissue homogenates, supplemented with NADPH and arachidonic acid [20:4(n-6)] as a substrate. During the incubation and the extraction, most of the 5,6-epoxyeicosatrienoic acid (5,6-EpETriE) was converted to 5,6-dihydroxyeicosatrienoic acid (5,6-DiHETriE), and most of the 5,6-DiHETriE was converted to 5,6-DiHETriE-delta-lactone. Consequently, the chief degradation product of 5,6-EpETriE and 5,6-DiHETriE in the incubation mixture was 5,6-DiHETriE-delta-lactone. 5,6-DiHETriE-delta-lactone, corresponding to [20:4(n-6)], was shown to be characterized by a high intensity of quasimolecular ions (MH+ and MNH4+), using ion analysis obtained by reversed-phase HPLC-thermospray MS. On selected-ion monitoring (SIM) chromatograms of 5,6-DiHETriE-delta-lactone and with deuterium-labeled 15(S)-hydroxyeicosatetraenoic acid as the internal standard, the regression equation of the peak-area ratio and the amount of 5,6-DiHETriE-delta-lactone was y = 12.2x + 0.7 (r = 0.9996). 5,6-Epoxygenase activity was represented as the sum of the amount of 5,6-DiHETriE-delta-lactone, 5,6-EpETriE and 5,6-DiHETriE per mg protein, after 30 min in an incubation mixture. The activity from rat brain homogenate decreased considerably with growth of the rat.
We have purified 3'-phosphoadenosine-5'-phosphosulfate:GalCer sulfotransferase [EC 2.8.2.11] from a human renal cancer cell line SMKT-R3 through a combination of affinity chromatographies using galactosylsphingosine, 3',5'-bisphosphoadenosine and heparin as ligands. The purified sulfotransferase showed a specific activity of 1.2 mumol/min/mg, which is 300 times more than the highest activity among the enzyme preparations purified so far from other sources. Homogeneity of the purified sulfotransferase was supported by the facts that the enzyme preparation showed a single protein band with an apparent molecular mass of 54 kDa on reducing SDS-PAGE and that protein bands coincided with the enzyme activity on both native PAGE and nonreducing SDS-PAGE. GalCer was the best acceptor for the purified enzyme. LacCer, GalAAG, and GalDG were also good acceptors. GlcCer, Gg3Cer, Gg4Cer, Gb4Cer, and nLc4Cer did serve as acceptors although the relative activities were low. On the other hand, the enzyme could not act on Gb3Cer, which possesses alpha-galactoside at the nonreducing terminus. Neither galactose nor lactose served as an acceptor. These observations suggest that the sulfotransferase prefers beta-glycoside, especially beta-galactoside, at the nonreducing termini of sugar chains attached to a lipid moiety.
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Cholesteryl ester transfer protein (CETP) plays an important role in regulating the concentration and composition of high density lipoprotein (HDL) and low density lipoprotein (LDL). Although several genetic abnormalities causing CETP deficiency have been identified in the Japanese subjects with a marked hyperalphalipoproteinemia (HALP), there are many CETP-deficient subjects for whom the genetic abnormalities have not been clarified. In the present study, we analyzed the molecular basis of an HALP subject without CETP activity and mass, and found a novel mutation in the CETP gene. This novel mutation (G181X) was a G-to-T substitution at codon 181 of exon 6 which replaced a codon for glycine (GGA) with a premature stop codon (TGA). The G181X mutation created a new cutting site by restriction enzyme MaeIII. To estimate the frequency of G181X, we investigated unrelated 294 HALP (HDL-cholesterol > or = 2.59 mmol/L = 100 mg/dl) subjects by restriction fragment length polymorphism (RFLP) analysis with Mae III. One (0.34%) HALP subject was homozygous and four (1.36%) were heterozygous for this mutation. The allelic frequency of a G-to-T substitution at codon 181 of exon 6 was 0.0102 in HALP subjects. From the lipid analysis of the proband and the homozygote, it was clarified that the G181X mutation had dominant effects on HDL and LDL metabolism, similar to a G-to-A substitution at the 5' splice donor site of the intron 14 (1451 + 1G-->A). In conclusion, the G181X mutation is one of causes of HALP in the Japanese HALP subjects, having dominant effects on lipid metabolism.
OBJECTIVE: We have reported that ventromedial hypothalamic (VMH) lesions induced marked hyperglycemia and a distinct reduction in pancreatic insulin content during short-term observation in male Goto-Kakizaki (GK) rats, an animal model for non-insulin-dependent diabetes mellitus (NIDDM) (Metabolism 43: 32-37, 1994). We investigated the long-term effect of VMH lesions on glucose metabolism, pancreatic insulin content, abdominal fat distribution and vascular complications in male GK rats. DESIGN: Metabolic and histological examinations in male GK rats during 16 weeks after making VMH lesions were compared to those in sham operated GK or Wistar rats. SUBJECTS: Eleven 9-week-old male GK rats and 4 male Wistar rats. VMH-lesions were made in 6 GK rats and sham operation were performed on 5 GK rats and 4 Wistar rats as controls. MEASUREMENTS: Food intake, body weight, and plasma glucose, insulin and lipid levels at 2 weeks interval after operation. Urinary protein and albumin levels at 15 weeks after operation. Measurement of pancreatic insulin content, mesenteric fat and abdominal subcutaneous fat weights, and histological examinations of kidney and aorta were performed after 16 weeks. RESULTS: Although food intake increased in VMH-lesioned GK (GK-VMH) rats compared with that in sham-operated GK (GK-sham) rats, the body weight of GK-VMH rats was significantly less than that of GK-sham rats. Plasma glucose was markedly elevated in GK-VMH rats from 2 through 16 weeks after operation, while it was only mildly increased in GK-sham rats. Plasma insulin levels were higher in GK-VMH rats one week after operation and thereafter tended to be lower compared to those in GK-sham rats. Plasma triglyceride levels were significantly increased in GK-VMH rats. The insulin content of pancreas at 16 weeks after operation was markedly decreased in GK-VMH rats. VMH lesions caused a significant 1.2-fold increase in mesenteric fat weight and a 1.3-fold higher ratio of mesenteric fat weight to subcutaneous fat weight in GK rats compared with sham-operated rats at 16 weeks after operation. The urinary excretions of protein and albumin in GK-VMH rats were greater than those in GK-sham rats. Histological examinations of the kidneys in GK-VMH rats revealed that the glomerular basement membranes were thicker than those of GK-sham rats. The descending aorta in GK-VMH rats also showed morphologic changes in the intima characteristic of an early stage of atherosclerosis. CONCLUSION: Male GK-VMH rats may be a useful animal model for non-obese NIDDM with visceral fat accumulation, which develops typical diabetic complications, including both microangiopathy and macroangiopathy.
The homogenate of a brain or liver obtained from a 1-55-day-old rat was incubated with NADPH and docosahexaenoic or arachidonic acid as the substrate. omega-Hydroxydocosahexaenoic or omega-hydroxyeicosatetraenoic acid from an incubation mixture of the homogenate was detected on a selected-ion monitoring chromatogram of reversed phase-HPLC-thermospray-mass spectrometry. omega-Hydroxylation activity in the brain homogenate considerably increased with growth up to 55 days. Activity in the liver homogenate decreased much with growth up to 55 days. omega-Hydroxylation activity in homogenates of rat brain gray matter, white matter, medula oblongata and cerebellum was much the same. omega-Hydroxylation activity of docosahexaenoic acid in rat brain homogenate was maximal at pH 7.5-8.0 in 50 mM Tris-HCl buffer and was inhibited by CO gas, metyrapone, ADP-Fe3+, heat treatment at 100 degrees C for 5 min and without NADPH. Based on these results, it is suggested that omega-hydroxylation activity is associated with cytochrome P-450 without NADPH-ADP-Fe(3+)-dependent lipid peroxidation, and the omega-hydroxylation system may be a metabolic pathway of the fatty acids in adult rat brain or neonatal rat liver. Since omega-hydroxyeicosatetraenoic acid produces relaxation of artery, it is suggested that blood flow changes in rat brain or liver with growth are caused by omega-hydroxylation activity changes in these organs with growth.
Human colonic well differentiated adenocarcinoma homogenate was incubated with NADPH and docosahexaenoic acid (22:6(n-3)) or arachidonic acid (20:4(n-6)) as a substrate. On a selected ion monitoring chromatogram obtained with reversed phase-high-performance liquid chromatography thermospray mass spectrometry, omega-hydroxydocosahexaenoic acid (omega-HDHE) or omega-hydroxyeicosatetraenoic acid (omega-HETE) from an incubation mixture of the homogenate was detected in significant amount, compared to that from a colonic region remote from the carcinoma. In contrast, epoxydocosapentaenoic acids and the dihydroxy derivatives from 22:6(n-3) or epoxyeicosatrienoic acids and the dihydroxy derivatives from 20:4(n-6) were detected in low amounts, compared to that from a colonic region remote from the carcinoma. The results suggest that highly active NADPH-dependent omega-oxidations of polyunsaturated fatty acids occur in colonic adenocarcinoma homogenate.
A method for the analysis of omega-carboxyleukotriene B4 and omega-hydroxyleukotriene B4 in human colonic carcinoma homogenate is described. The hydroxy groups of the leukotriene metabolite were acetylated by acetic anhydride, and the mixture was partially purified on a Sep-Pak C18 cartridge and analysed by reversed-phase HPLC-thermospray MS. Generally, the base ion, [MH-2(60)]+, is produced through elimination of two acetic acid (60 mass units) molecules from the protonated molecular ion. On selected-ion monitoring, standard curves for omega-carboxy- or omega-hydroxyleukotriene B4 showed a linear relationship over the range 72-1500 pmol. The assay based on selected-ion monitoring was applied to an extract from human colonic carcinoma homogenate. When a homogenate of human colonic well-differentiated adenocarcinoma was incubated with NADPH and leukotriene B4 (60.6 nmol) as a substrate, the conversion of precursor leukotriene B4 to omega-carboxyleukotriene B4 or omega-hydroxyleukotriene B4 was 0.33 or 3.17%, respectively. Based on these results, it is suggested that carcinoma cells themselves or leukocytes at the hostsite in a region of human colonic well-differentiated adenocarcinoma are performing omega-oxidation through NADPH-dependent omega-hydroxylation of leukotriene B4.
We have recently identified a novel 40-kDa heatshock protein hsp40 as a mammalian homologue of bacterial DnaJ protein. Here we demonstrate the physical interaction between hsp70 (DnaK homologue) and hsp40 in human cells as determined by immunoprecipitation methods. Co-immunoprecipitation of hsp70 with hsp40 was dependent on the presence of ATP or unfolded protein (reduced carboxymethylated alpha-lactalbumin). A mutant type of tumor suppressor gene product, mtp53, was co-immunoprecipitated not only with hsp70 but also with hsp40. These results suggest the existence of a hsp70(DnaK)/hsp40(DnaJ) chaperone system in mammalian cells.
A novel 40-kDa heat-shock protein hsp 40 in mammalian cells has been recently identified to be a homolog of bacterial DnaJ protein. We have previously shown the colocalization of hsc70 (p73, constitutive form) with hsp40 in the nucleoli of heat-shocked HeLa cells. In this report we further investigated intracellular translocation and localization of hsp40 and hsp70 (both constitutive p73 and inducible p72) in several mammalian cells. Translocation kinetics of hsp40 during heating at mild temperature were almost the same as those of hsp70 in HeLa cells. Hsp40 colocalized not only with hsc70 (p73) but also with hsp70 (p72) in heat-shocked HeLa (human), HA-1 (Chinese hamster) and NRK (rat) cells. Direct interaction of hsp40 with hsp70 (p73 and/or p72) was observed in all cells tested by immunoprecipitation methods. Also, treatments of cells with cytoskeleton-acting drugs such as cytochalasin E, colchicine and taxol had no effect on the heat-induced translocation of hsc70 (p73) and hsp40 in NRK cells. These results strongly suggest that hsp40 and hsp70 (p73/p72) form a complex in the cytoplasm at normal temperature, translocate together and colocalize in the nuclei and nucleoli upon heat-shock, and that they may function cooperatively to repair (refold) denatured proteins under stress conditions.
We report chronological changes of neuro-otological findings and associated neuropathological findings in 2 patients. Concerning gaze limitation, upward gaze was primarily disturbed, followed by downward gaze and abduction. Adduction tended to be preserved although convergence was disturbed at the early stage. And, upward gaze limitation was followed by loss of oculocephalic responses (OCRs). Bell's phenomenon was preserved until the late stage of the disease. Caloric nystagmus was absent at the early stage. One patient showed dissociation between nystagmus and vestibulo-collic reflex in the caloric test at the early stage of the disease. Neuropathological examination revealed gliosis in the interstitial nucleus of Cajal and the MLF as well as neuronal loss and gliosis in the oculomotor nuclei. The vestibular nuclei also showed gliosis and atrophy of nerve cells.
We report two cases of successful surgical treatment for Stanford type A aortic dissection with thrombosed false lumen and saccular aneurysm formation. They were admitted to our clinic for severe anterior chest and back pain. Diagnosis was made as a Stanford type A dissection with thrombosed false lumen by chest CT, aortography and cardiac ultrasonography. Aortography showed ulcer like projection (ulp) in the ascending aorta, although flow in false lumen was not visualized. They were initially treated with antihypertensive drugs. Six and eight weeks later, although medical therapy was continued, a saccular aneurysm of the ascending aorta was found to be increased in size rapidly in both cases. Operation performed was the ascending aorta replacement with resection of the saccular aneurysm. The location of the ulp was found to be corresponded with the intimal tear at surgery. The postoperative courses were uneventful. Thus, careful medical follow up and surgical treatment were necessary in these cases. We believe that the ulp represents defect in the intima leading to the clotted false lumen and provides a possible sign of rupture in clinical course of aortic dissection.
To assess the alteration of apolipoprotein (apo) B mRNA editing in non-insulin-dependent diabetes mellitus (NIDDM), we measured plasma apoB-100 and apoB-48 levels and apoB mRNA editing efficiency in the liver and intestine from GK (Goto-Kakizaki) rats, a genetically NIDDM animal. Male GK rats and control littermates, aged 25 weeks, were used in this study. Ventromedial hypothalamus (VMH)-lesioned control rats were used as hyperinsulinemic models. VMH-lesioned GK rats (GK+VMH) were treated as an insulin-exhausted NIDDM model. Plasma cholesterol and triglyceride levels were increased in GK rats. Very low density lipoprotein (VLDL)-triglyceride and low density lipoprotein (LDL)-cholesterol concentrations were significantly higher in GK rats than in controls. The increase of VLDL-triglyceride was most marked in GK+VMH rats. Plasma apoB-48 levels, quantified by immunoblot, were increased in GK rats. However, apoB-100 levels were minimally elevated in GK rats. Therefore, the apoB-48/apoB-100 ratio was remarkably increased in GK rats. ApoB mRNA editing was analyzed by reverse transcriptase-polymerase chain reaction coupled with dideoxynucleotide chain termination assay. The ratio of apoB-48-type cDNA to apoB-100-type cDNA was significantly increased in the liver from GK rats compared with controls. Although the development of the VMH lesion increased plasma apoB-48 levels, it had no effect on the proportion of apoB-48-type to apoB-100-type cDNA in the liver from both GK and control littermates. ApoB mRNA in the intestine was almost totally edited (approximately 95%). Intestinal apoB-48/apoB-100 cDNA ratio showed no significant difference among the four groups. In conclusion, an enhanced apoB mRNA editing was indicated in the non-insulin-dependent diabetic rats, which might contribute to the increase of plasma apoB-48 levels.
We recently experienced a case of infective endocarditis with multiple embolic complications. The patient was a 21-year-old male who was admitted to our hospital with high fever and dyspnea. Echocardiography showed vegetation on the aortic valve, severe aortic regurgitation and mild mitral regurgitation. Abdominal echography revealed large splenic infarction. First we performed aortic valve replacement and splenectomy. But infective endocarditis developed after surgery, and echocardiography revealed severe mitral regurgitation, which has been a change for the worse. After treatment with antibiotics, mitral valve replacement (the second surgery) was performed. Intraoperative findings were confirmative of subtotal rupture of the posterior papillary muscle. Histological examination of the subtotal ruptured muscle revealed presence of coagulation necrosis. The patient is healthy without recurrence of infective endocarditis after the second surgery. It is exceedingly rare to experience a case of surgically treated subtotal papillary muscle rupture due to infective endocarditis.
We presented a case of a 48-year-old male with failed delivery of the Palmaz-Schatz stent, who subsequently required CABG with successful retrieval of the stent. The patient had presented recurrent angina, approximately one and a half year since the last PTCA to the LAD. Re-PTCA was performed, but resulted in a severe local dissection. The stent implantation was tried, but was inadvertently dropped in the vessel. Since the patient was hemodynamically stable, we tried to remove the stent with Retriever endovascular snare, which ended up with interlocking the stent in the more proximal segment of the LAD and the snare itself was not removed. We, therefore, resort to the emergent operation, with successful retrieval of the snare device and the stent and carried on to the the bypass operation to the distal LAD without any complications. To our best knowledge, this is the first report in Japan for surgical retrieval of the Palmaz-Schatz stent, which was failed in delivery and interlocked in the vessel with retriever device.