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T Ikenoue

Publications and source records attributed to T Ikenoue.

At least 55 records · Page 3Linked to original sources
PubMed1999

Heat shock protein 72 expression and microtubule-associated protein 2 disappearance after hypoxia-ischemia in the developing rat brain.

OBJECTIVE: This study was intended to investigate the temporal changes in heat shock protein 72 expression and microtubule-associated protein 2 disappearance in rat brain at 2 different ages after hypoxic-ischemic insult. STUDY DESIGN: Both 5-day-old and 14-day-old Wistar rats were subjected to unilateral common carotid artery ligation and hypoxia in 8% oxygen for 2 hours at 33 degrees C. Brain sections were examined sequentially for heat shock protein 72 expression at 0.5, 3, 6, 12, 24, 48, and 72 hours of recovery after hypoxia-ischemia and for microtubule-associated protein 2 disappearance at 0, 24, 48, and 72 hours of recovery and at 7 days of recovery after hypoxia-ischemia. Results of immunohistochemical staining for heat shock protein 72 and microtubule-associated protein 2 were used as markers for detection of early hypoxic-ischemic brain damage. Permanent neuronal damage was assessed with hematoxylin and eosin staining at 7 days after hypoxia. RESULTS: In 5-day-old rats microtubule-associated protein 2 expression was lost as early as 0 hours after hypoxia-ischemia in the cerebral cortex and hippocampus, with a peak at 48 hours after which expression recovered. Expression of heat shock protein 72 was detected in the ligated hemisphere at 0.5 hours after hypoxia-ischemia and peaked at 6 to 24 hours of recovery. In 14-day-old rats microtubule-associated protein 2 was stained in the cortex at 0 hours after hypoxia-ischemia but gradually disappeared in the cerebral cortex and hippocampus after 24 hours of recovery. The expression of heat shock protein 72 was not detected by 6 hours of recovery in the cerebral cortex and by 3 to 12 hours of recovery in the hippocampus, but heat shock protein 72 was persistently expressed in the cortex and hippocampus after 48 hours of recovery. Neuronal damage was significantly less in 5-day-old rats than in 14-day-old rats. CONCLUSION: In 5-day-old rats hypoxia-ischemia causes earlier changes in heat shock protein 72 and microtubule-associated protein 2 immunostaining results and causes less severe brain damage than in 14-day-old rats.

Aging↗
PubMed1999

Expression of glial cell line-derived neurotrophic factor in the brain and cerebrospinal fluid of the developing rat.

Expression of GDNF in developing rat brain from PND 1 to 14 and on PND 21 was examined immunocytochemically. At PND 1, intense diffuse immunoreactivity was noted within the cytoplasm of a diverse group of neuronal and nonneuronal cells, including choroid plexus epithelial cells, ependymal cells, tanycytes of the third ventricle, and cellular elements in the subarachnoid compartment. GDNF expression became more localized among these cells from PND 7-14 and was almost undetectable by PND 21. Although GDNF-positive small glial cells were scattered within the cerebral cortical plate and the striatum already at PND 1, GDNF expression among astroglial cells within the corpus callosum and in the white matter adjacent to the lateral ventricles was more prominent between PND 5 and 8. GDNF expression among the pyramidal neurons of the cerebral cortex was evident relatively early in the postnatal period, but the neurons of the hippocampus and thalamus showed more intense immunoreactivity at later periods between PND 8-14. ELISA of the CSF revealed a rapid rise in GDNF levels from 71.4 +/- 10.9 pg/ml (mean +/- S.E.M.) at PND 1 to peak levels of 138.4 +/- 18.5, 135.1 +/- 5.4 and 132.9 +/- 8.0 pg/ml, at PND 5, 7 and 9, respectively. Peak CSF levels of GDNF occurred when GDNF expression was intense within astroglial cells in the corpus callosum and cerebral white matter. Thereafter, the levels gradually decreased to 76.5 +/- 9.7 pg/ml at PND 21. The widespread expression of GDNF among different cellular elements in the developing brain suggest that GDNF probably plays diverse functional roles in many different neuronal systems in addition to its known effects on the dopaminergic system. Developmental shifts in GDNF expression further suggest that GDNF may be of critical importance at different stages of brain growth and differentiation.

Aging↗
PubMed1999

[Clinicobacteriological studies on the nine cases with upper genital tract Mycoplasma hominis infection].

Clinicobacteriological characteristics of nine cases isolated Mycoplasma hominis from the genital tract were studied, and the following results were obtained: elevation of IgG antibodies to M. hominis was measured by ELISA in all cases, but in the MI method only one case showed an elevation of metabolic inhibitory antibody. Convalescent sera from seven patients showed additional and high density bands which were not recognized by acute phase sera in immunoblotting. It was thought that in two patients M. hominis was a causal bacteria for pelvic inflammatory disease (PID). In three cases, it was suggested that M. hominis was related to a premature delivery and idiopathic labor. As infectious symptoms, two patients had body temperatures of more than 38 degrees C but other cases showed 37-37.8 degrees C. Though all cases showed an elevation of CRP, six elevations were slight. As a medication beta-lactam agents were administrated, but their efficacy was not recognized. Furthermore, two patients showed spontaneous recovery in spite of improper antimicrobial agents administration or drainage combined with antimicrobial agents. From the above results. It was thought that M. hominis played a causative role in upper genital tract infection.

Adult↗
PubMed1999

Structure of cag pathogenicity island in Japanese Helicobacter pylori isolates.

BACKGROUND: cag pathogenicity island (PAI) is reported to be a major virulence factor of Helicobacter pylori. AIM: To characterise cagA and the cag PAI in Japanese H pylori strains. METHODS: H pylori isolates from Japanese patients were evaluated for CagA by immunoblot, for cagA transcription by northern blot, and for cagA and 13 other cag PAI genes by Southern blot. cagA negative strains from Western countries were also studied. Induction of interleukin-8 secretion from gastric epithelial cells was also investigated. RESULTS: All Japanese strains retained cagA. Fifty nine of 63 (94%) strains had all the cag PAI genes. In the remaining four, cag PAI was partially deleted, lacking cagA transcripts and not producing CagA protein. Details of the PAI of these strains were checked; three lacked cagB to cagQ (cagI) and continuously cagS to cag13 (cagII), and the remaining one lacked cagB to cag8. Western cagA negative strains completely lacked cag PAI including cagA. Nucleotide sequence analysis in one strain in which the cag PAI was partially deleted showed that the partial deletion contained 25 kb of cag PAI and the cagA promoter. Interleukin-8 induction was lower with the cag PAI partial deletion strains than with the intact ones. All Japanese cag PAI deleted strains were derived from patients with non-ulcer dyspepsia, whereas 41 of 59 (70%) CagA-producing strains were from patients with peptic ulcers or gastric cancer (p<0.05). CONCLUSIONS: Most Japanese H pylori strains had the intact cag PAI. However, some lacked most of the cag PAI in spite of the presence of cagA. Thus the presence of the cagA gene is not an invariable marker of cag PAI related virulence in Japanese strains.

Adult↗
PubMed1999

Adenovirus mediated p53 tumour suppressor gene therapy for human gastric cancer cells in vitro and in vivo.

BACKGROUND/AIMS: Gastric cancer is one of the most prevalent forms of cancer in East Asia. Point mutation of the p53 gene has been reported in more than 60% of cases of gastric cancer and can lead to genetic instability and uncontrolled cell proliferation. The purpose of this investigation was to evaluate the potential of p53 gene therapy for gastric cancer. METHODS: The responses of human gastric cancer cell lines, MKN1, MKN7, MKN28, MKN45, and TMK-1, to recombinant adenoviruses encoding wild type p53 (AdCAp53) were analysed in vitro. The efficacy of the AdCAp53 treatment for MKN1 and MKN45 subcutaneous tumours in nude mice was assessed in vivo. RESULTS: p53-specific growth inhibition was observed in vitro in two of four gastric cancer cell lines with mutated p53, but not in the wild type p53 cell line. The mechanism of the killing of gastric cancer cells by AdCAp53 was found, by flow cytometric analysis and detection of DNA fragmentation, to be apoptosis. In vivo studies showed that the growth of subcutaneous tumours of p53 mutant MKN1 cells was significantly inhibited by direct injection of AdCAp53, but no significant growth inhibition was detected in the growth of p53 wild type MKN45 tumours. CONCLUSIONS: Adenovirus mediated reintroduction of wild type p53 is a potential clinical utility in gene therapy for gastric cancers.

Adenoviridae↗
PubMed1999

Expression of insulin-like growth factor I messenger ribonucleic acid in developing osteophytes in murine experimental osteoarthritis and in rats inoculated with growth hormone-secreting tumor.

Osteophytes are one of the characteristic features of osteoarthritis and are often found in acromegalic arthropathy. The aim of this study was to investigate insulin-like growth factor I (IGF-I) involvement in osteophyte formation. One percent collagenase solution was injected into murine knee joints as an osteoarthritis model. In a different animal group, GH-secreting tumor cells were inoculated s.c. to the rat thigh as an acromegaly model. A series of osteophyte formation was examined histologically. IGF-I messenger RNA was detected using the in situ hybridization method. Type I IGF receptors were detected immunohistochemically. In the osteoarthritis model, osteophyte formation appeared as synovial or perichondral cell proliferation adjacent to the articular cartilage on day 5, followed by cartilage formation on day 7 and endochondral ossification on day 14. In the acromegaly model, synovial or perichondral cell proliferation was observed 4 weeks after inoculation, followed by osteophyte formation at 8 weeks. In both models, IGF-I messenger RNA and type I IGF receptor were coexpressed by proliferating synovial or perichondral cells, proliferating chondrocytes, and osteoblasts within the developing osteophytes. These results suggest that IGF-I regulated the initiation and development of osteophyte formation in both models in an autocrine and/or paracrine fashion.

Animals↗
PubMed1999

Posterior tibial slope in the normal and varus knee.

Posterior tibial slope was evaluated in 30 normal and 30 varus knees using magnetic resonance imaging. The mean tibial posterior slope in the medial plateau was 10.7 degrees (range: 5 degrees - 15.5 degrees) in the normal knees and 9.9 degrees (range: 1.5 degrees - 19 degrees) in the varus knees. The mean tibial posterior slope in the lateral plateau was 7.2 degrees (range: 0 degrees - 14.5 degrees) in the normal knees and 6 degrees (range: 1 degrees - 13 degrees) in the varus knees. Although these differences were not statistically significant, there was a large range in the posterior tibial slope in both groups. When 10-mm thickness of bone was resected in the middle of the lateral plateau in total knee arthroplasty, the thickness of the bone resection in the anterior and posterior part of the lateral plateau was measured. The presumed thickness of bone was 10.1 mm (range: 7.3-13.9 mm) in the anterior part and 7.5 mm (range: 5-10.3 mm) in the posterior part. A large range in the measured thickness of bone also was detected in the varus knees. These results suggest that the shape of the proximal tibia varies among individuals and that the cutting angle should be determined in each case to avoid any mismatch of the patient's posterior slope of the tibia after implantation.

Case-Control Studies↗
PubMed1999

[Comparison of the effect of rapid infusion of lactated and that of acetated Ringer's solutions on maternal and fetal metabolism and acid-base balance].

The maternal and neonatal metabolism and acid-base balance were investigated in 20 parturients undergoing combined spinal and epidural anesthesia for cesarean delivery. Patients received intravenous infusion at a rate of either 25 ml.kg-1.h-1 of lactated (LR group, n = 10) or acetated (AR group, n = 10) Ringer's solution before anesthesia, to prevent hypotension during anesthesia. We obtained venous blood samples as follows; maternal control before anesthesia, maternal sample A after the infusion, umbilical sample B, and neonatal pedal sample C 5 h after birth, and determined lactate, pyruvate, bicarbonate, and base excess concentrations, and pH in each sample. In sample A, the lactate level was significantly higher and base excess level was significantly lower in the LR group than in the AR group. The pH of sample A and B was significantly higher in the AR group than in the LR group. However, no differences in all parameters of sample C between the two groups were observed. These results demonstrated that acetated Ringer's solution is better than lactated Ringer's solution in rapid infusion before cesarean section because of the correction of neonatal lactic acidosis.

Acid-Base Equilibrium↗
PubMed1998

Charged amino acids at the carboxyl-terminal portions determine the intracellular locations of two isoforms of cytochrome b5.

Outer mitochondrial membrane cytochrome b5 (OMb), which is an isoform of cytochrome b5 (cyt b5) in the endoplasmic reticulum, is a typical tail-anchored protein of the outer mitochondrial membrane. We cloned cDNA containing the complete amino acid sequence of OMb and found that the protein has no typical structural feature common to the mitochondrial targeting signal at the amino terminus. To identify the region responsible for the mitochondrial targeting of OMb, various mutated proteins were expressed in cultured mammalian cells, and the subcellular localization of the expressed proteins was analyzed. The deletion of more than 11 amino acid residues from the carboxyl-terminal end of OMb abolished the targeting of the protein to the mitochondria. When the carboxyl-terminal 10 amino acids of OMb were fused to the cyt b5 that was previously deleted in the corresponding 10 residues, the fused protein localized in the mitochondria, thereby indicating that the carboxyl-terminal 10 amino acid residues of OMb have sufficient information to transport OMb to the mitochondria. The replacement of either of the two positively charged residues within the carboxyl-terminal 10 amino acids by alanine resulted in the transport of the mutant proteins to the endoplasmic reticulum. The mutant cyt b5, in which the acidic amino acid in its carboxyl-terminal end was replaced by basic amino acid, could be transported to the mitochondria. It would thus seem that charged amino acids in the carboxyl-terminal portion of these proteins determine their locations in the cell.

Amino Acid Sequence↗
PubMed1998

Hypoxic-ischemic tolerance phenomenon observed in neonatal rat brain.

OBJECTIVE: Our purpose was to determine whether hypoxic-ischemic brain damage would be protected against by advance conditioning of the animal with 4 hours of hypoxic exposure. STUDY DESIGN: Neonatal rats were exposed on postnatal day 7 to (1) 4 hours of hypoxia with 8% oxygen (preconditioning hypoxic group) or (2) 4 hours of normoxia (sham-preconditioning group). At 24 hours after the conditioning, rats from both groups were exposed to left carotid artery ligation followed by 2 hours of hypoxia (8% oxygen/92% nitrogen). All the rats were killed 1 week after hypoxia-ischemia, and their brains were extracted for histologic study. RESULTS: Two types of brain damage were histologically observed at 1 week after hypoxia-ischemia in both groups: (1) generalized infarction in the ligated hemisphere and (2) predominant neuronal loss in the hippocampal region. The total incidence of brain damage was significantly decreased in the preconditioning hypoxic group (10/24 rats, 41.7%) compared with the sham-preconditioning hypoxic group (17/22 rats, 77.3%; P < .05). CONCLUSION: Our results show that the hypoxic-ischemic tolerance phenomenon may be induced in the hypoxic-ischemic brain damage model by conditioning the animal before the insult with 4 hours of hypoxic exposure.

Animals↗
PubMed1998

Interleukin-8 production in primary cultures of human gastric epithelial cells induced by Helicobacter pylori.

Interleukin-8 (IL-8) production by the gastric mucosa is increased in Helicobacter pylori infection. Previous studies indicated that H. pylori induces IL-8 synthesis in cancer cell lines, and the ability of H. pylori to stimulate IL-8 production is supposed to be associated with cagA and other cag pathogenicity island genes, including picB gene. In the present study, we investigated the induction of IL-8 in primary cultures of normal human gastric epithelial cells to elucidate the IL-8 induction by wild type strains and by the picB knockout strain. Human gastric epithelial cells were obtained from surgically resected specimens from four patients. Three H. pylori strains (TN2F4; type 1 clinical isolate, TN2F4m1; isogenic picB mutant of TN2F4, Tx30a; type 2 strain) were cocultured with the normal gastric epithelial cells or the transformed MKN-28. IL-8 levels in culture medium were determined by enzyme immunoassay. Human gastric epithelial cells produced IL-8 at a 10-50 times higher level than MKN-28 did when cocultured with TN2F4. The mutant TN2F4m1 induced IL-8 at significantly lower levels than the parent strain. Cells from four patients behaved similarly on IL-8 production. The results of the present study demonstrated the induction of IL-8 in normal gastric epithelial cells, suggesting that picB gene product may play an essential role in vivo.

Bacterial Proteins↗
PubMed1998

Effects of 4 hours magnesium sulfate infusion on fetal heart rate variability and reactivity in a goat model.

Effects of magnesium sulfate were investigated on fetal heart rate (FHR) baseline, variability, and reactivity in goats. Six chronically catheterized fetuses of Japanese Saanen goat at 125 to 130 days' gestation (term = 147 days) were used. Magnesium sulfate was directly infused to the fetuses. Short-term variability and long-term variability were obtained according to Huey et al. The baseline, reactivity, short-term variability and long-term variability of the FHR were compared between those receiving magnesium sulfate infusions and those receiving vehicle infusions without magnesium sulfate for 4 hr. Two-way analysis of variance (ANOVA) and Duncan's multiple range test was applied for statistical significance. Four hours magnesium sulfate infusion significantly increased fetal plasma concentration of magnesium from 2.4-6.6 mg/dL, without significant changes in fetal respiratory gases and pH values. The baseline FHR was significantly decreased by magnesium infusion compared with that receiving vehicle infusion. The incidence of acceleration, short-term variability, and long-term variability during the fourth hour of magnesium infusion was also significantly decreased compared to a controlled infusion. The time spent by high amplitude phase of short-term variability and that of long-term variability were also significantly reduced. Significant correlation was obtained between the magnesium concentration and incidence of acceleration at fourth hour of magnesium infusion. Four hours infusion of magnesium sulfate significantly decreases baseline FHR, short-term variability, long-term variability, and reactivity in fetal goats at 0.85 gestation.

Animals↗
PubMed1998

Two patients with premature labour associated with Mycoplasma hominis infection.

Because several reports have suggested that bacterial vaginosis causes premature labour and early rupture of the fetal membranes, the presence of a bacterial flora that causes bacterial vaginosis is thought to be a risk factor for premature labour. The present study investigated two patients with premature delivery and intra-uterine Mycoplasma hominis infection. In microbiological studies, Gram's staining of amniotic fluids revealed numerous neutrophils and epithelial cells but no micro-organisms. Culture of amniotic fluid before antibiotic therapy yielded only M. hominis under anaerobic conditions; aerobic culture was negative. Vaginal discharge taken on the day of delivery yielded no growth in case 1 and M. hominis and Enterococcus faecalis in case 2. Maternal sera showed specific antibodies to M. hominis by ELISA and immunoblotting. As no possible cause of premature labour other than M. hominis infection was detected, it is concluded that the intra-uterine M. hominis infection was associated with premature labour in these patients.

Adult↗
PubMed1998

Major virulence factors, VacA and CagA, are commonly positive in Helicobacter pylori isolates in Japan.

BACKGROUND: VacA and CagA proteins have been reported to be major virulence factors of Helicobacter pylori. However, antibodies against these proteins are frequently found in the sera of Japanese patients regardless of their gastroduodenal status. AIM: To evaluate the expression of VacA and CagA proteins by H pylori strains isolated in Japan. METHODS: By using specific antibodies raised against recombinant VacA and CagA proteins, the expression of VacA and CagA was evaluated in 68 H pylori strains isolated from Japanese patients; a vacuolating assay and genotyping of the vacA gene were also used in the evaluation. The results in analysed in relation to the gastroduodenal diseases of the hosts. RESULTS: VacA and CagA proteins were expressed in 59/68 (87%) and in 61/68 (90%) isolates respectively. The vacuolating assay was positive in 57/68 (84%) isolates, indicating that most immunologically VacA positive strains produced active cytotoxin. The prevalence of infection with strains expressing CagA and positive for vacuolating activity (Type I) was very high, 54/68 (79%), irrespective of the gastroduodenal status of the host. CONCLUSION: Most H pylori isolates in Japan are positive for vacuolating cytotoxin and CagA, and thus these virulence factors cannot be used as markers to discern the risk of developing serious gastroduodenal pathologies in the hosts. However, the high prevalence of infection with strains positive for vacuolating cytotoxin and CagA may contribute to the characteristics of H pylori infection in Japan.

Adult↗
PubMed1998

Serum 1alpha,25-dihydroxyvitamin D3 accumulates into the fracture callus during rat femoral fracture healing.

1,25-dihydroxyvitamin D3 (1,25(OH)2D3) is thought to be an important systemic factor in the fracture repair process, but the mechanism of action of 1,25(OH)2D3 has not been clearly defined. In this study, the role of 1,25(OH)2D3 in the fracture repair process was analyzed in a rat closed femoral fracture model. The plasma concentration of 1,25(OH)2D3 rapidly decreased on day 3 and continued to decrease to 10 days after fracture. We assessed whether this decrease was based on the accelerated degradation or retardation of the synthesis rate of 1,25(OH)2D3, from 25(OH)D3. After radiolabeled 3H-1,25(OH)2D3 or 3H-25(OH)D3 was injected i.v. into fractured or control (unfractured) rats, the concentrations of 25(OH)D3 and 1,25(OH)2D3 metabolites were measured by HPLC. The plasma concentrations of these radiolabeled metabolites in fractured group were similar to those in control rats early after operation. However, radioactivity in the femurs of fractured rats was higher than that of the control group. Furthermore, the radioactivity was concentrated in the callus of the fractured group analyzed by autoradiography. 1,25(OH)2D3 receptor gene expression was detected early after fracture and, additionally, both in the soft and hard callus on days 7 and 13 after fracture. These results showed that the rapid disappearance of 1,25(OH)2D3 in the early stages after fracture was not due to either increased degradation or decreased synthesis of 1,25(OH)2D3, but rather to increased consumption. Further, these results suggest the possibility that plasma 1,25(OH)2D3 becomes localized in the callus and may regulate cellular events in the process of fracture healing.

Animals↗
PubMed1997

Sequence of neuronal responses assessed by immunohistochemistry in the newborn rat brain after hypoxia-ischemia.

OBJECTIVE: Our purpose was to study the neuronal responses of heat shock protein-72 (a stress-inducible protein) and microtubule-associated protein-2 (a constitutive protein of the neuronal cytoskeleton) after hypoxia-ischemia and their relationship with permanent damage in the newborn rat brain. STUDY DESIGN: Seven-day-old rats were exposed to unilateral carotid artery ligation followed by 2 hours of hypoxia (8% oxygen/92% nitrogen) and then killed at time points ranging from 1 to 72 hours after injury. Brains were removed for immunohistochemical and routine staining. RESULTS: Heat shock protein-72 appearance and microtubule-associated protein-2 disappearance occurred from 1 hour after injury, mainly in the dentate gyrus of the hippocampal formation and the cerebral cortex. Such alterations reached maximal levels at 24 hours for both proteins. Microtubule-associated protein-2 staining recovered in almost all parts of the brain. However, the hippocampal CA3 showed a delay in the responses for both proteins, and microtubule-associated protein-2 did not recover the response to immunostaining. Histologic evaluation at 72 hours after hypoxia by routine methods showed predominant damage in the hippocampal CA3. CONCLUSION: Our results show that delayed responses of heat shock protein-72 and microtubule-associated protein-2 are related to a high incidence of neuronal cell loss in the hippocampal CA3 region.

Animals↗
PubMed1997

High seropositivity of anti-CagA antibody in Helicobacter pylori-infected patients irrelevant to peptic ulcers and normal mucosa in Japan.

CagA-positive H. pylori is reported to be associated with gastroduodenal disease in Western countries. To evaluate the relationship between CagA and disease, cloning of the entire cagA gene (3771 bp), insertion of a partial fragment (1272 bp) into an expression vector, purification of the recombinant protein, production of an antibody against the recombinant CagA protein through rabbits, and use of the recombinant CagA protein as an antigen, detection of the anti-CagA antibody by western blotting were all performed. Sera of 132 H. pylori-infected patients undergoing endoscopy were studied. Anti-CagA antibodies were detected in 90%, 87%, 90%, 94%, and 93% of patients with gastric ulcer (N = 34), duodenal ulcer (N = 27), chronic gastritis (N = 31), gastric cancer (N = 17), and normal mucosa (N = 15), respectively. High seropositivity of anti-CagA antibody even in individuals with normal mucosa indicated that CagA may not be a unique marker for disease by H. pylori infection in Japan.

Amino Acid Sequence↗