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Biomedical subjects

T Shimazu

Publications and source records attributed to T Shimazu.

At least 91 records · Page 5Linked to original sources

Two regions of the Mn-stabilizing protein from Synechococcus elongatus that are involved in binding to photosystem II complexes.

Limited proteolysis of the Mn-stabilizing protein (MSP) from the thermophilic cyanobacterium Synechococcus elongatus with chymotrypsin, trypsin or lysylendopeptidase that yielded four major polypeptides of 26 kDa, 22 kDa, 15 kDa and 11 kDa on denaturing gel electrophoresis resulted in total loss of the binding capacity of the protein to PSII complexes. Analyses of electrophoretic patterns and amino-terminal sequences of the proteolytic products revealed that the three proteases specifically cleaved the protein at a site between Phe156 and Gly163 or between Arg184 and Ser191. Site-directed mutagenesis was used to construct two mutant MSPs that had a nick between Phe156 and Leu157, a chymotrypsin-cleavage site, and Met before Leu157 or in place of Leu157. The two mutant proteins failed to bind to PSII complexes, although they largely retained ordered secondary structure and comigrated with the wild-type proteins in non-denaturing gel electrophoresis. The loss of the protein binding can be ascribed to introduction of a nick because a mutant protein that had Met in place of Leu157 but no nick was able to specifically bind to the functional site of PSII complexes and restore the oxygen-evolving activity as effectively as the wild-type protein. In contrast, a mutant MSP with Met inserted between Phe156 and Leu157 bound only weakly and non-specifically to PSII complexes and failed to reactivate oxygen evolution. Thus, the binding of the protein to the functional site of the PSII complex was highly sensitive to a small structural change that was caused by cleavage or insertion of a single amino acid residue between Phe156 and Leu157. The results suggest that the Phe156-Gly163 and Arg184-Ser191 sequences of the cyanobacterial MSP are regions for interaction with PSII complexes.

Amino Acid Sequence↗

Effects of noradrenaline on the cell-surface glucose transporters in cultured brown adipocytes: novel mechanism for selective activation of GLUT1 glucose transporters.

Glucose transport into rat brown adipocytes has been shown to be stimulated directly by the sympathetic neurotransmitter, noradrenaline, without a significant increase in the protein content of either GLUT1 or GLUT4 glucose transporter in the plasma membrane [Shimizu, Kielar, Minokoshi and Shimazu (1996) Biochem. J. 314, 485-490]. In the present study, we labelled the exofacial glucose-binding sites of GLUT1 and GLUT4 with a membrane-impermeant photoaffinity reagent, 2-N-[4-(1-azitrifluoroethyl)benzoyl]-[2-3H]1,3-bis- (D-mannos-4-yloxy)-2-propylamine (ATB-[3H]BMPA), to determine which isoform is responsible for the noradrenaline-induced increase in glucose transport into intact brown adipocytes in culture. Insulin stimulated the rate of hexose transport by increasing ATB-[3H]BMPA-labelled cell-surface GLUT4. In contrast, the noradrenaline-induced increase in glucose transport was not accompanied by an increased ATB-[3H]BMPA labelling of GLUT4, nor with an increased amount of GLUT4 in the plasma membrane fraction as assessed by Western blotting, indicating that noradrenaline does not promote the translocation of GLUT4. However, noradrenaline induced an increase in photoaffinity labelling of cell-surface GLUT1 without an apparent increase in the immunoreactive GLUT1 protein in the plasma membrane. This is suggestive of an increased affinity of GLUT1 for the ligand. In fact, the Ki value of non-radioactive ATB-BMPA for 2-deoxy-D-glucose uptake was significantly decreased after treatment of the cells with noradrenaline. The increased photoaffinity labelling of GLUT1 and increased glucose transport caused by noradrenaline were inhibited by a cAMP antagonist, cAMP-S Rp-isomer. These results demonstrate that noradrenaline stimulates glucose transport in brown adipocytes by enhancing the functional activity of GLUT1 through a cAMP-dependent mechanism.

Adipose Tissue, Brown↗

Endothelin 1 aggravates acute liver injury in perfused livers of rats after treatment with D-galactosamine.

The effects of endothelin 1 (ET-1) on hemodynamics and acute liver damage were studied using perfused livers of rats treated with D-galactosamine. In control liver perfused in situ with constant pressure, infusion of ET-1 into the portal vein at a concentration of 0.1 nmol/L decreased the flow rate without a significant leakage of lactate dehydrogenase (LDH) or aspartate transaminase (AST) into the effluent. In contrast, in similarly perfused liver 24 hours after treatment with D-galactosamine (800 mg/kg intraperitoneally), ET-1 caused rapid and remarkable increases in the leakage of LDH and AST from the liver accompanied by the reduction of perfusion flow to the extent similar to that observed in control livers. In addition, ET-1 decreased oxygen uptake and bile secretion in galactosamine-treated livers. The potentiating effects of ET-1 on enzyme leakage were also observed under constant flow conditions. Moreover, infusion of the thromboxane A2 analogue at a concentration of 10 nmol/L decreased the flow rate markedly, yet the rapid increases in enzyme leakage were not observed. Infusion of ET-3 induced the responses of flow reduction and the potentiation of rapid enzyme leakage similar to those obtained with ET-1. Neither the endothelin A-receptor antagonist BQ485 nor the endothelin B-receptor antagonist BQ788 could inhibit the acute liver damage caused by ET-1; instead they exaggerated its effects. The combination of both antagonists together, however, almost completely suppressed the flow reduction and the potentiation of enzyme leakage caused by ET-1. These results indicate that ET-1 is capable of aggravating acute liver damage not merely through reduction of the flow rate but through direct action on liver cells. They also suggest that both the endothelin A and endothelin B receptors are involved in this action of ET-1.

Acute Disease↗

Profiles of circulating inflammatory- and anti-inflammatory cytokines in patients with hemolytic uremic syndrome due to E. coli O157 infection.

The systemic inflammatory response to Escherichia coli O157 infection was studied from the profiles of circulating inflammatory and anti-inflammatory cytokines. Twelve patients transferred sequentially to our hospital for the intensive care with acute illness due to Escherichia coli O157 infection and the possible form of haemolytic uraemic syndrome were included in this study. Increased circulating concentrations of tumour necrosis factor, interleukin 6, interleukin 8, granulocyte colony-stimulating factor, and interleukin 10 were found in patients with various stages of this infection and haemolytic uraemic syndrome. Especially, the degree of the increase of circulating interleukin 10 in those who had a typical signs of haemolytic uraemic syndrome was higher than those of other inflammatory cytokines. Two groups of E. coli infection could be classified into one with a typical haemolytic uraemic syndrome and the other with atypically bacteremic state over haemolytic uraemic syndrome according to these cytokine levels.

Adult↗

Noradrenaline and ATP decrease the secretion of triglyceride and apoprotein B from perfused rat liver.

To explore the role of hepatic sympathetic nerves on the secretion of very low density lipoprotein (VLDL), the effects of sympathetic neurotransmitters, noradrenaline and ATP, on the secretion of triglyceride and apoprotein B (ApoB) from the liver were studied using rat liver perfused in situ with recirculation. During liver perfusion with physiological medium, the amount of triglyceride in the perfusate was increased linearly for up to 120 min. The addition of noradrenaline to the perfusion medium at a final concentration of 1 muM increased the portal pressure and suppressed the secretion of triglyceride to about 60% of control without an increase in free fatty acid production. The administration of ATP increased the portal pressure with kinetics different from those induced by noradrenaline, but suppressed the triglyceride secretion to an extent similar to that induced by noradrenaline. The suppressive effect of noradrenaline on triglyceride secretion was mimicked by an alpha-adrenoceptor agonist, phenylephrine, and was retained after the haemodynamic changes were prevented by sodium nitroprusside. The secretion of ApoB from the perfused liver was also inhibited by noradrenaline or ATP to about 70% of control. However, hepatic levels of mRNA for ApoB were not significantly altered by noradrenaline and ATP. Since ApoB is the major apoprotein in VLDL, these results suggest that the sympathetic neurotransmitters noradrenaline and ATP suppress the secretion of ApoB-containing lipoprotein including VLDL from the liver, probably acting on post-transcriptional processes.

Adenosine Triphosphate↗

Post-transcriptional control of the level of mRNA by hepatitis B virus X gene in the transient expression system using human hepatic cells.

BACKGROUND: Hepatitis B virus (HBV) infection is closely related to the development of not only acute or chronic hepatitis, but also hepatocellular carcinoma. Among the HBV genes, the X gene has been implicated in the carcinogenicity of this virus as a major causative factor by its ability to activate viral and cellular genes in trans via protein-protein interaction with cellular factors without binding to DNA. RESULTS: To explore the possibility of other functions of the X gene, we examined the effect of X protein on the transient expression system of simian virus 40 (SV40) large T-antigen or chloramphenicol acetyltransferase (CAT) mRNA using SV40 promoter or EF-1alpha (human elongation factor 1alpha) promoter, by co-transfecting an X gene expression plasmid to human hepatic cell lines, HepG2 and Huh7. In contrast to the SV40 promoter-mediated expression, the level of both T-antigen and CAT mRNAs expressed from the EF-1alpha promoter was strikingly decreased by X protein in both hepatic cells. The nuclear run-on assay and the mRNA decay experiment using actinomycin D, indicated that the effect of X protein on the lowering of the level of chimeric mRNA was due to the degradation of mRNA, but not repression of transcriptional initiation. Moreover, this effect was dependent on the 22 bp sequence in the 5' untranslated region of mRNA derived from the EF-1alpha promoter. CONCLUSION: The present data suggest a new function of the X gene to post-transcriptionally control the stability of mRNA through the 5' untranslated region derived from the EF-1alpha promoter in human hepatic cells.

Antigens, Polyomavirus Transforming↗

Recombinant human granulocyte colony-stimulating factor attenuates inflammatory responses in septic patients with neutropenia.

OBJECTIVE: The objective of this study was to determine the effects of recombinant human granulocyte colony-stimulating factor (rhG-CSF) administration in septic patients with neutropenia. METHODS: Twenty consecutive septic patients were administered rhG-CSF subcutaneously (2 microg x kg(-1) x d(-1)) for 5 days (group G). They were compared with 14 septic patients treated earlier without rhG-CSF (group N). All patients in both groups met the criteria of total leukocyte count (TLC) less than 5,000/mm3 and C-reactive protein (CRP) more than 10 mg/dL. Changes in TLC, absolute neutrophil count (ANC), CRP, respiratory index (RI), Acute Physiology and Chronic Health Evaluation (APACHE) II score, and Goris's Multiple Organ Failure (MOF) index were evaluated. In addition, nucleated cell count (NCC), differentiation in bone marrow aspiration, neutrophil phagocytic and bactericidal activity, serum concentrations of interleukin-6 (IL-6) and IL-8 as inflammatory markers, and plasma concentration of leukocyte elastase (LE) as an indicator of the tissue injury were evaluated in group G. RESULTS: In group G, TLC, ANC, NCC, and neutrophil functions increased significantly, whereas CRP, IL-6, and IL-8 decreased reciprocally. There was no deterioration of LE and RI. Consequently, the APACHE II score and MOF index improved. In group N, however, CRP showed no change concomitant with the APACHE II score and MOF index. CONCLUSION: Administration of rhG-CSF attenuates inflammatory responses without inducing tissue injury in septic patients with neutropenia.

APACHE↗

Aggravating action of zymosan on acute liver damage in perfused liver of rats treated with D-galactosamine.

To study the role of Kupffer cells in the aggravation of liver injury, effects of zymosan on acute liver damage were explored using perfused livers of rats 24 h after intraperitoneal injection of D-galactosamine (800 mg/kg). The leakage of lactate dehydrogenase and aspartate aminotransferase into the effluent was used to indicate acute liver damage. Infusion of zymosan (30 microgram/ml) into the portal vein rapidly increased the leakage of lactate dehydrogenase and aspartate aminotransferase from galactosamine-treated liver with decreased perfusion flow. Pretreatment of animals with gadolinium, which diminished an immunostaining of resident macrophages in the injured liver, significantly attenuated the flow reduction induced by zymosan, whereas it did not affect the increases in enzyme leakage. Infusions of PGF2alpha, PGE2, and leukotriene D4, the eicosanoids mainly produced by Kupffer cells, decreased perfusion flow without rapid augmentation of enzyme leakage from galactosamine-treated liver. These results indicate that zymosan potentiates acute liver damage after galactosamine injection and suggest that certain types of nonparenchymal cells other than Kupffer cells are mainly involved in the action of zymosan.

Acute Disease↗

A poison information service via an automated facsimile (fax) system: an adjunct to the operator-based service.

BACKGROUND: Poison centers are faced with the escalating costs of specialist staffing and increased investments in hardware and databases despite deficit funding. We developed an automated fax information system to access poison information from any fax machine without special training or equipment. METHODS: We provided a 3-month trial service of the fax system in conjunction with the regular operator-based service and analyzed the fax access log, followed by a questionnaire to the 2204 affiliate members regarding the use of the fax. RESULTS: A total 657 accesses to the fax system were made, of which 105 (16%) were unsuccessful; 342 (52%) were made to retrieve the user's manual, 85 (13%) to retrieve the index pages, and 230 (35%) to retrieve documentation on specific substances. The most frequently accessed items concerned disc battery ingestion (13.5%), salicylates (10.3%), mamushi viper (7.1%), acetaminophen (5.8%), and sodium hypochlorite (3.8%). The questionnaires were returned by 666 (30.2%) members; 93 (14%) had actually used the fax system with the average frequency of 1.8 times/user, 63% (59/93) of the respondents considered the service satisfactory, and 33% (31/93) said it was somewhat unsatisfactory. CONCLUSIONS: The automated fax information system was accepted and handled by users with only minor difficulty. A facsimile information service may have a valuable role in providing poisoning information and has potential benefits in cases of environmental disasters.

Consumer Behavior↗

Selection of severely head injured patients for mild hypothermia therapy.

OBJECT: The authors have analyzed the efficacy of inducing mild hypothermia (34 degrees C) in 62 severely head injured patients to control fulminant intracranial hypertension. METHODS: All 62 patients fulfilled the following criteria: 1)persistent intracranial pressure (ICP) greater than 20 mm Hg despite fluid restriction, hyperventilation, and high-dose barbiturate therapy; 2) an ICP lower than the mean arterial pressure; and 3) a Glasgow Coma Scale (GCS) score of 8 or less on admission. The patients were divided into three groups based on computerized tomography findings: extracerebral hematoma (34 patients with subdural and/or epidural hematoma), focal cerebral lesion (20 patients with localized brain contusion and/or intracerebral hematoma), and diffuse swelling (eight patients with no focal mass lesion). Mild hypothermia prevented ICP elevation in 35 (56.5%) of the 62 patients whose ICP was greater than 20 mm Hg despite conventional therapies. Among those 35 patients whose ICP was controlled by mild hypothermia, 12 (34.3%) achieved functional recovery (good outcome or moderate disability). However, functional recovery was observed in only five (10.9%) of the 46 patients whose ICP was greater than 40 mm Hg after conventional therapies. Of 40 patients with an admission GCS score of 5 to 8, there were 11 (27.5%) who achieved functional recovery. On the contrary, mild hypothermia was not effective in 22 patients with an admission GCS score of 3 or 4. In the patients with focal cerebral lesions, ICP was controlled by mild hypothermia in 17 patients (85%) and patient outcome was intimately related to the extent of the damage. Among 18 patients with extracerebral hematoma who had a midline shift of 9 to 12 mm, raised ICP could be successfully controlled by mild hypothermia in 16 patients (88.9%) and three (16.7%) achieved functional recovery. However, ICP could not be controlled in patients with extracerebral hematoma who had a midline shift of 13 mm or more. In patients with diffuse swelling, ICP elevation could not be prevented at all by mild hypothermia. CONCLUSIONS: The authors conclude that mild hypothermia is effective for preventing ICP elevation in patients without diffuse brain swelling in whom ICP remains higher than 20 mm Hg but less than 40 mm Hg after conventional therapies.

Adolescent↗

[Clinical and pathophysiologic problems associated with smoke inhalation injury].

Smoke inhalation injury is one of the primary determinants of survival following major burn injury. The primary site of injury in smoke inhalation appears to be the small airway rather than the alveoli, and thus small airway occlusion caused by edema and pseudomembrane formation are the primary mechanisms of progressive hypoxia. Ventilation-perfusion (VA/Q) alterations after smoke inhalation are characterized by increased blood flow to low VA/Q compartments, although an increase in true shunt (VA/Q = 0) was not a consistent finding. This differs considerably from most adult respiratory distress syndrome (ARDS) patients or oleic acid-induced lung edema models, in which an increase in true shunt is the major mechanism of hypoxia. Such differences lead to different responses to nitric oxide (NO) inhalation therapy, and NO does not improve oxygenation and outcome in patients with smoke inhalation injury. In the treatment of inhalation injury, meticulous removal of pseudomembrane by fiberoptic bronchoscopy is essential, the use of high concentrations of oxygen should be avoided since it can cause absorption atelectasis. High-frequency percussive ventilation is a suitable treatment for inhalation injury, as it improves oxygenation and facilitates removal of pseudomembrane.

Burns, Inhalation↗

[Hypouricemia in patients with meningitis].

Serum urate and sodium concentrations were measured in 23 patients with acute viral and bacterial meningitis. Serum urate level was 3.0 +/- 0.2 mg/dl (mean +/- S.D.) (3.6 +/- 1.2 mg/dl in male and 2.5 +/- 0.9 mg /dl in female) on admission, but gradually elevated with improvements of meningitis. It turned to 4.8 +/- 0.2 mg/dl after recovery, and the value on admission was significantly lower than that after recovery (p < 0.0001). Serum sodium level was 137.6 +/- 2.9 mEq/l on admission and 139.7 +/- 2.7 mEq/l after recovery; also lower in the former (p < 0.01). These results show that patients develop transient hypouricemia, which may be explained by SIADH (syndrome of inappropriate secretion of ADH), although SIADH is subclinical in most cases of meningitis.

Adult↗

Transcriptional activation of the human c-myc gene by simian virus 40 large T antigen without binding to p53 and RB proteins in the transient expression system.

Transcriptional activation of the human c-myc gene by SV40 large T antigen was examined using HepG2 cells by co-transfecting a T antigen expression plasmid with a myc-CAT construct containing the 2.3-kb upstream region from the P1 promoter and the P2 promoter region fused to the CAT gene. T antigen increased the basal activity of the P2 promoter region containing the E2F binding site, but both the P2 promoter region and the upstream region from the P1 promoter were important for overall activation by T antigen. CAT assay using mutated T antigen lacking p53 or the RB binding site indicated that p53 or RB was not mainly involved in transcriptional activation of the c-myc gene. It appears that activation of the c-myc gene by T antigen is probably dependent upon E2F and a cellular factor through a mechanism which is independent of binding of T antigen to p53 and RB.

Antigens, Polyomavirus Transforming↗

Paradoxical positive nitrogen balance in burn patients receiving high-dose administration of insulin for nutritional care.

BACKGROUND: Nitrogen balance in patients who need high-dose administration of insulin has not been evaluated clinically. The purpose of this study was to compare the difference in nitrogen balance between burn patients who received high-dose administration of insulin and those who did not. METHODS: This study was performed in 19 severely burned adults with no liver or kidney failure. Patients were divided into two groups on the basis of the mean ratio of administered insulin and calorie intake (I/C) for the initial 4 weeks, a high I/C group (n = 9) and a low I/C group (n = 10). There were no significant differences between the two groups regarding age, percentage of area burned, and body weight. Nitrogen balance, blood urea nitrogen, and urine urea nitrogen were measured in all patients. Plasma concentrations of glucose, insulin, glucagon, cortisol, and urinary excretion of 3-methyl-histidine were measured in 12 patients (six in each group). RESULTS: Until day 10 both groups exhibited similar changes in plasma concentrations of glucose, insulin, glucagon, and cortisol. Subsequently, plasma concentrations of insulin and glucagon began to decrease in the low I/C group, whereas a high level was sustained in the high I/C group (p < 0.05). Plasma glucose and cortisol measurements showed no significant differences between the two groups. Blood urea nitrogen levels and urinary excretion of 3-methyl-histidine were not different between the two groups. Urine urea nitrogen excretion in the high I/C group, however, was significantly lower than that in the low I/C group from day 8 (p < 0.05). Thus the high I/C group achieved positive nitrogen balance more quickly than the low I/C group. Paradoxically, however, the high I/C group was at higher risk of septic complications and exhibited higher mortality than the low I/C group (p < 0.05). CONCLUSIONS: These results indicate that an improvement in nitrogen balance, which is accepted as a good thing in the management of critically ill patients, is not necessarily good in the high I/C group and that residual nitrogen was retained within the body in the high I/C group.

Adult↗