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Biomedical subjects

T Itoh

Publications and source records attributed to T Itoh.

At least 145 records · Page 8Linked to original sources

In vivo and in vitro analyses of the AmyR binding site of the Aspergillus nidulans agdA promoter; requirement of the CGG direct repeat for induction and high affinity binding of AmyR.

The alpha-glucosidase gene (agdA) of Aspergillus nidulans has a single CGGN8CGG type AmyR binding site in its promoter region. The binding site is functional in vivo as a cis-element responsible for induction by starch, and mutational studies indicated that both the CGG triplets are required for high-level induction. A part of AmyR (residues 1-411; AmyR(1-411)), which was produced as a MalE fusion protein in E. coli, bound to the CGGN8CGG site of the agdA promoter. DNA binding profiles to the mutant binding sites that lacked both or either one of the CGG triplets suggested that AmyR(1-411) can bind to a single CGG triplet site with low affinity and that two AmyR molecules cooperatively bind to the CGG direct repeat.

Aspergillus nidulans↗

Clinical usefulness of duplex ultrasonography for the assessment of renal arteriosclerosis in essential hypertensive patients.

The present study was carried out to investigate whether the renal resistive index (RRI), obtained by ultrasonic duplex scanning, is useful for the evaluation of renal arteriosclerosis in essential hypertensive patients. We also studied the relationships between RRI and other kinds of hypertensive target-organ damage, including carotid atherosclerosis. One hundred and two patients (56.4+/-9.4 years) with untreated mild or moderate essential hypertension were examined. The normal range of RRI was determined for 12 normal age-matched volunteers (55.0+/-6.6 years). Hypertensive organ damage was evaluated by funduscopy, electrocardiograms, and carotid B-mode imaging. Based on the mean and distribution of RRI in normal volunteers (0.60+/-0.05), the normal upper limit of RRI was found to be 0.7. RRI was correlated with creatinine clearance (CCr) (r=-0.61, p<0.05), and blood urea nitrogen (r=0.46, p<0.05), but not with serum creatinine. In addition, the incidence of abnormal RRI (>0.7) was higher in patients with left ventricular hypertrophy and in those with advanced carotid atherosclerosis (p<0.01, respectively). Thus, RRI appears to be more strongly associated with CCr than with serum creatinine, and it increases in patients with hypertensive end-organ damage. The assessment of RRI may be useful for the evaluation of early renal damage in essential hypertension.

Adult↗

Comparison of bacteriological, genetic and pathological characters between Escherichia coli O115a,c:K(B) and Citrobacter rodentium.

Murine pathogenic Escherichia coli O115a,c:K(B) (MPEC) is the causative agent of mouse megaenteron, the pathology of which resembles that of transmissible murine colonic hyperplasia caused by Citrobacter rodentium. We compared their genetic and pathological features to reveal the relationship between these two bacteria. To evaluate the genetic distances, 16S rDNA genes were sequenced and biochemical reactions were tested. Mouse strain susceptibility tests, using CF1 MPEC-susceptible germfree mice and BALB/cA(Jic) resistant mice were performed. MPEC strains and C. rodentium showed more than 99.6% identity by comparison of 16S rDNA gene sequences. All results from biochemical reactions and the mouse strain susceptibility tests were identical. It is proposed that MPEC should be reclassified as C. rodentium.

Animals↗

Microbiological contamination in genetically modified animals and proposals for a microbiological test standard for national universities in Japan.

The Biosafety Committee of the Japanese Association of Laboratory Animal Facilities of National Universities (JALAN) investigated recent episodes of microbiological contamination in genetically modified mice (GMM), and the countermeasures taken when the contaminated GMM were introduced into animal facilities, by questionnaires addressed to 53 animal facilities belonging to JALAN and serological tests. Although almost all of the contaminated GMM were accepted with conditions such as rederivation after or before reception and housing in designated rooms, contamination with a spectrum of microorganisms was demonstrated in GMM transferred domestically and from abroad. In serological tests, Mycoplasma pulmonis, mouse parvovirus, and mouse encephalomylitis virus were detected in GMM transferred from domestic facilities and from abroad. The present results of the questionnaires and serological tests suggest that GMM are highly and widely contaminated with microorganisms compared with mice from commercial breeders. Thus, we propose a microbiological requirement, including microbiological status--excellent, common, and minimum--as a guide for the transfer and procurement of mice and rats in Japan.

Animals↗

A possible relationship between abortions and placental embolism in pregnant rabbits given human granulocyte colony-stimulating factor.

Developmental toxicity studies were conducted in rats and rabbits with a human G-CSF derivative (NTG). As reported for G-CSF, increases in abortions and fetal mortality were observed in rabbits, but not in rats given NTG. Histopathological examination of the rabbit placenta revealed accumulation of neutrophils in vessels and necrosis of the tissues surrounding these vessels. To assess the mechanism of abortion and fetal death in rabbits given G-CSF, 125I-labeled NTG was given intravenously on Day 18 of pregnancy after repeated administration of cold NTG on Days 6 through 17 of pregnancy, and the feto-maternal distribution of radioactivity was examined. In a rabbit given 20 micrograms/kg, high radioactivity was observed in the endometrium, placenta, and some parts of the decidua at 6 hr when the concentration of radioactivity in maternal blood had already decreased. At 24 hr after administration of 200 micrograms/kg NTG, high radioactivity was still detected in parts of the maternal placenta. These patterns of distribution suggest that embolism occurred in parts of the uterus and placenta which might have caused congestion. Radioactivity in the TCA precipitates in the fetus was low, suggesting that NTG does not readily transfer to the fetus. These results strongly suggest that neutrophils accumulated in the vessels of placenta and induced embolism leading to abortions and fetal mortality in the rabbits given G-CSF.

Absorption↗

Role of blood volume in the age-associated decline in peak oxygen uptake in humans.

It has been reported that maximal oxygen uptake (VO(2 max)) is linearly correlated with blood volume (BV) in young people and that there is a reduction in VO(2 max) with aging. To examine the involvement of BV in the reduction of VO(2 max), we used an incremental cycle ergometer protocol in a semi-recumbent position to determine the relationship between peak oxygen uptake (VO(2 peak)) and BV in older subjects (69.1 +/- 1.0 years; n = 22), then compared that relationship with that in young subjects (22.3 +/- 0.5 years; n = 31). In the present study, VO(2 peak) and BV were significantly lower in the older subjects, compared with those in the young subjects. A linear correlation was demonstrated between the VO(2 peak) and BV in both the older (r = 0.705; p < 0.001) and the young (r = 0.681; p < 0.001) subjects within the groups. However, an analysis of covariance with BV as a covariate revealed that VO(2 peak) at a given BV was smaller in the older subjects than in the young subjects (p < 0.001), i.e., graphically, the regression line determined for the older subjects showed a downward shift. The decreased peak heart rate as a result of aging (153 +/- 3 beats/min in the older vs. 189 +/- 2 beats/min in the young subjects) contributed partly to this downward shift. These results suggest that the BV is an important determinant factor for VO(2 peak), especially within an age group, and that the age-associated decline of VO(2 peak) is also, to a relatively larger degree, because of factors other than BV and heart rate.

Adult↗

Trauma sites and clinical features associated with acute hyperextension spinal cord injury without bone damage--relationship between trauma site and severity.

To elucidate whether a relationship exists between the site of trauma and severity of acute hyperextension spinal cord injury without bone damage, we examined the clinical features of 25 male and 10 female patients aged 13 to 88 years. None of the patients had vertebral damage such as fracture and dislocation. The site of impact was classified as the buccal, forehead, or mandibular region. The neurological findings were assessed according to Frankel's classification at admission and at follow up after 3 months or more to assess outcome. Eleven patients suffered trauma in the buccal region, one patient in Frankel's grade B, three in grade C, and seven in grade D at admission. All 11 of these patients showed an improvement of one grade or more to an outcome of C in one patient, D in one, and E in nine. Trauma occurred at the forehead region in 18 patients, four in grade B, 10 in grade C, and four in grade D. Improvement was seen at follow up by one grade or more to C in one patient, D in 10, and E in seven. Trauma occurred at the mandibular region in six patients, four in grade B and two in grade C. Four of these patients showed improvement of one grade or more to grade B in one, grade C in four, and grade E in one. Overall, seven patients had poor outcomes, five of whom suffered trauma to the mandibular region, indicating that impact to the mandibular region tends to have an unfavorable clinical outcome. Our findings indicate that the site of trauma greatly influences the severity of hyperextension spinal cord injury.

Adolescent↗

Effects of long-term administration of recombinant bovine tumor necrosis factor-alpha on glucose metabolism and growth hormone secretion in steers.

OBJECTIVE: To investigate the effects of long-term administration of recombinant bovine tumor necrosis factor-alpha (rbTNF) on plasma glucose and growth hormone concentrations, and to determine whether treatment with rbTNF causes insulin resistance in steers. ANIMALS: 5 steers treated with rbTNF and 5 steers treated with saline (0.9% NaCl) solution (control). PROCEDURES: In experiment 1, rbTNF (5.0 microg/kg of body weight) or saline solution (5 ml) was administered SC daily for 12 days. Blood samples were obtained before treatment, and plasma was harvested for determination of glucose, insulin, and growth hormone (GH) concentrations. In experiment 2, insulin, glucose, or growth hormone-releasing hormone (GHRH) was administered IV on days 7, 9, and 11, respectively, after initiation of rbTNF or saline treatment in experiment 1. Plasma glucose and insulin concentrations were measured before and at various times for 4 hours after insulin or glucose administration. Plasma GH concentrations were measured at various times for 3 hours after GHRH administration. RESULTS: In experiment 1, administration of rbTNF resulted in hyperinsulinemia without hypoglycemia and decreased plasma GH concentrations. In experiment 2, plasma glucose concentrations were higher in steers treated with rbTNF and insulin than in controls. Plasma GH concentrations were lower in steers treated with rbTNF and GHRH than in controls. CONCLUSIONS AND CLINICAL RELEVANCE: Prolonged treatment with rbTNF induced insulin resistance and inhibited GHRH-stimulated release of GH in steers. Results indicate that rbTNF is a proximal mediator of insulin resistance and inhibits release of GH during periods of endotoxemia or infection.

Animals↗

Establishment of a mouse thymic epithelial cell line, IT-76MHC and a brief review on cultured thymic epithelial cells.

Interactions between thymocytes and thymic stromal cells are essential for thymocyte differentiation, but little evidence has been presented to directly show in vivo functions or interactions of the stromal cells. Among the stromal cells, the thymic epithelial cell has been considered to have profound effect on thymocyte differentiation and maturation. The calcium-depleted medium, originally developed for the culture of mouse epidermal cells, was applied for the culture of the mouse thymic epithelial cells, and successfully, an epithelial cell line, IT-76MHC was obtained from the mouse thymus. IT-76MHC cells were identified as distinct mouse thymic epithelial cells by 1/ mosaic-like arrangement, 2/ presence of well-developed desmosome and 3/ tonofilaments, 4/ positivity for cytokeratin, and 5/ induced expression of MHC class I and II by IFN-gamma treatment. IGF-1, IGF-2, oxytocin and vasopressin were also detected immunohistochemically in IT-76MHC cells. Furthermore, the IT-76MHC thymic epithelial cells, when injected intrathymically in the allogeneic mouse, prolonged the survival of skin graft from the same donor strain that IT-76MHC cells were derived. These results demonstrate that the thymic epithelial cell line IT-76MHC produces modest thymocyte survival factors as well as a growth suppressor, and that IT-76MHC cells have the ability to induce transplantation tolerance probably through their expression of MHC class I and II molecules. Taken altogether, the IT-76MHC thymic epithelial cells have been proved to be useful tools to better understand the in vivo functions of thymic epithelial cells, and to gain a deep insight into their involvement in the critical selection process of thymocytes which still remains obscure. Finally and additionally, literatures so far reported on thymic epithelial cells in culture, especially lines and clones, are reviewed and their identity as well as their functions are discussed.

Animals↗

Polyamines of the hyperthermophilic archaebacteria belonging to the genera Thermococcus and Methanothermus and two new genera Caldivirga and Palaeococcus.

Cellular polyamines of eight new thermophilic archaebacteria were investigated to determine the chemotaxonomic significance of polyamine distribution profiles. Hyperthermoacidophilic Caldivirga maquilingensis belonging to the family Thermoproteaceae of the Crenarchaeota have a unique polyamine profile comprising spermidine, norspermidine and norspermine as the major polyamines. Within the order Thermococcales of the Euryarchaeota, the major polyamines of an extremely thermophilic terrestrial species of Thermococcus, T. zilligii, were spermidine and agmatine, whereas hyperthermophilic submarine species of Thermococcus and hyperthermophilic submarine Palaeococcus ferrophilus contained a quaternary branched penta-amine, N4-bis(aminopropyl)spermidine, as a major polyamine. A hyperthermophilic methanogen, Methanothermus sociabilis, belonging to Euryarchaeota, contained spermidine and spermine as the major polyamine.

Polyamines↗

Polyamines of the thermophilic eubacteria belonging to the genera Thermosipho, Thermaerobacter and Caldicellulosiruptor.

Cellular polyamines of four new thermophiles located in three early branched eubacterial clades, were investigated for the chemotaxonomic significance of polyamine distribution profiles. The thermophilic anaerobic Thermosipho japonicus, belonging to the order Thermotogales, contained norspermidine, norspermine and thermospermine in addition to spermidine and spermine. The polyamine profile was identical to the polyamine composition of Thermotoga, Fervidobacterium and Petrotoga species of the order. Spermidine, norspermidine, spermine, N4-bis(aminopropyl)spermidine and agmatine were found in thermophilic aerobic Thermaerobacter marianensis. Some differences were observed in the polyamine compositions of the phylogenetically related thermophilic anaerobes, Moorella, Dictyoglomus, Thermoanaerobacterium and Thermoanaerobacter species. Thermophilic anaerobic Caldicellulosiruptor kristianssonii and Caldicellulosiruptor owensensis contained a linear penta-amine, thermopentamine, and two quaternary branched penta-amines, N4-bis(aminopropyl)spermidine and N4-bis(aminopropyl)norspermidine, as the major polyamines. A novel tertiary branched penta-amine, N4-aminopropylspermine, was found in the two Caldicellulosiruptor species.

Bacteria↗

An oral drug delivery system targeting immune-regulating cells ameliorates mucosal injury in trinitrobenzene sulfonic acid-induced colitis.

Control of immune-regulating cells in the colonic mucosa is important in the treatment of patients with inflammatory bowel disease (IBD). The aim of study was to examine the therapeutic effect of dexamethasone (DX) microspheres on 2,4,6-trinitrobenzene sulfonic acid (TNBS)-induced colitis in rats, a model for human Crohn's disease. DX microspheres and DX alone were administered orally to rats with TNBS-induced colitis. The macroscopic score, histological score, myeloperoxidase (MPO) activity, nitric oxide (NO) production, and gene expressions of proinflammatory cytokines, cyclooxygenase (COX)-1, and COX-2 in the colonic tissue were determined. Proliferating cell nuclear antigen (PCNA) staining and expression of nuclear transcription factor (NF)-kappaB in colonic tissues were also investigated. Macroscopic score, histological score, MPO activity, and NO production in rats treated with DX microspheres were significantly lower than in those treated with DX alone. The gene expression of proinflammatory cytokines and COX-2 in rats treated with DX microspheres was down-regulated, compared with that in rats treated with DX alone. The number of PCNA-positive cells in the DX microsphere group was larger than in the group treated with DX alone. DX microspheres suppressed NF-kappaB activation in TNBS-induced colitis more strongly than DX alone. Oral administration of DX microspheres appears to ameliorate mucosal injury in TNBS-induced colitis. This drug delivery system could be an ideal therapy for human IBD.

Administration, Oral↗

Effect of bacterial metabolism in the intestine on colorectal tumors induced by 1,2-dimethylhydrazine in transgenic mice harboring human prototype c-Ha-ras genes.

The number of colorectal tumors per mouse induced by 1,2-dimethylhydrazine in transgenic (Tg) mice carrying human c-Ha-ras genes was significantly reduced by ingestion of apple pectin (AP) or a culture condensate of Bifidobacterium longum(MB) compared with a control diet and non-Tg mice. However, there were no differences in the composition of fecal flora, water content, beta-glucuronidase and beta-glucosidase activities, and concentrations of organic acids and putrefactive products in the feces between the AP or MB diet and the control diet, or between the Tg mice and non-Tg mice. The concentration of secondary bile acids in the MB diet group was higher than that in the control group. These results suggested that there was no relationship between prevention of colorectal tumors in Tg mice and the AP or MB diet, or improvement of the intestinal environment due to these functional foods.

9,10-Dimethyl-1,2-benzanthracene↗

Asymmetric synthesis of both enantiomers of point-difluorinated-eldanolide, analogues of insect sex pheromone.

Substitution of two fluorine atoms on organic molecules is expected to alter both chemical reactivity and biological activity due to the strong electron-withdrawing nature of fluorine. The synthesis of partly gem-difluorinated compounds remains a significant challenge to synthetic organic chemists. We report that [2,3]-Wittig rearrangement of 1,1,2-trifluoroallylic ether gave a new type of partly gem-difluorinated allylic alcohol: 6-methyl-4,4,5-trifluorohept-1,5-dien-3-ol, 3, in a highly stereoselective fashion, and optical resolution of alcohol 3 was accomplished via lipase PS(PCL)-catalyzed reaction. Using this alcohol as the starting material, the first asymmetric synthesis of both enantiomers of point-fluorinated-eldanolides, 2,2,5,5,6-pentafluoroeldanolide 1 and 5,5,6-trifluoroeldanolide 2, analogues of the sex pheromone of the male African sugarcane borer, has been demonstrated.

Alkadienes↗

[Development of the chromatography analysis of liquid trapping 99mTc-Technegas using Jikei impinger].

We developed the equipment which collected 99mTc-Technegas into liquid (saline) and named this equipment an impinger of Jikei University Style. We further developed a technique using this impinger to evaluate its formation qualitatively by paper chromatography. Utilizing this technique we investigated various factors that caused environmental contamination by 99mTc-Technegas, including changes over time in a Technegas generator. It was demonstrated that upon getting mixed with oxygen gas to the argon gas, the Technegas generator induced contamination of 99mTc-Pertechnegas easily, leading to changes in its formation. The change of formation quality of the Technegas generator with the lapse of time was also revealed. These findings indicated that the maintenance and inspection of the equipment were important. This method is a simple and easy technique for the evaluation of 99mTc-Technegas formation, making it possible to perform the quality control of examination agent and the Technegas generator.

Chromatography, Paper↗

[Bladder carcinoma producing granulocyte colony-stimulating factor (G-CSF): a case report].

An 84-year-old male was admitted to our hospital with complaints of leg edema and general fatigue. He had undergone transurethral resection of transitional cell bladder cancer, grade 3, pT1b, one year previously. The computed tomographic scan revealed a hypoattenuating bladder tumor, protruding extravesically. Laboratory examination showed remarkable leukocytosis of 46,900/mm3 in the peripheral blood and high value of granulocyte colony-stimulating factor (G-CSF) 226 pg/ml (normal: less than 30 pg/ml). The resection of the tumor (partial cystectomy) was performed. The histological diagnosis was transitional cell carcinoma, grade 3. The production of G-CSF was confirmed by immunohistochemical examination in the recurrent tumor and the surgical sample from the transurethral resection. The leukocyte count in the peripheral blood decreased to the normal range after surgery. But leukocytosis recurred one month postoperatively and the computed tomographic scan revealed intrapelvic tumor recurrence. He died due to drastic progression of recurrent tumor at three months postoperatively.

Aged↗

[Evaluation of efficiency of a multi-crystal scintillation camera Digirad 2020tc Imager using a solid-state detectors].

Digirad 2020tc Imager is the movable scintillation camera, consisting of combining multi-crystal scintillators (CsI(Tl)) and photo-diodes. Total numbers of element are 4096, which are further divided into 16 x 16 modules. Each module contains 4 x 4 elements. We have examined Digirad 2020tc according to NEMA (National Electrical Manufactures Association), and the following results are obtained; the maximum count rate; 221 kcps, total system uniformity; 1.3% (integral uniformity), 0.9% (differential uniformity), system spatial resolution; 6.97 +/- 0.72 mm (the LEHR collimator to 99mTc source at 10 cm), intrinsic energy resolution; 12.8%, total system sensitivity; 3270.8 cpm/MBq (with LEHR collimator using 99mTc source at 10 cm). Further more, we determined the contrast of an imaging using the pin-hole (100 microns phi) 99mTc source in order to know the signal per noise (S/N) ratio among the pixels (S/N; 93.4 +/- 46.2 (first pixels)). Although the physical dimension of the camera has a smaller field of view, comparing with the standard camera, Digirad 2020tc has the equivalent characteristics as well as that of the standard camera and its field view is enough to measure the adult lung perfusion using a diverging collimator. We will further examine Digirad 2020tc with its movable portability and expect applications in nuclear medicine.

Gamma Cameras↗

[Analysis of the action of dextran sulfate as a prophylaxis for peritoneal cancer metastasis, and its acting mechanism].

Although peritoneal cancer metastasis has the highest frequency of postoperative recurrence among digestive organ malignant tumors, there is no still decisive treatment. Dextran sulfate (DS) as a prophylaxis for cancer metastasis was examined with respect to its effect on cultured cells. DS was made to act on a strong adhesive neoplasm cell, and the action and acting mechanism were examined with respect to 1. readhesiveness, 2. cell cycle, and 3. gene analysis. The results suggest that: i) Once tumor cells are detached by DS, the free cells do not attach even when DS is removed, and ii) DS causes the cells to stop in the G1/G0 phase.

Animals↗