Biomedical subjects

M Nagao

Publications and source records attributed to M Nagao.

At least 127 records · Page 7Linked to original sources

PubMed1999

Autogenous onlay grafting for enhancement of extracortical tissue formation over porous-coated segmental replacement prostheses.

BACKGROUND: Prosthetic reconstruction with extracortical bone-bridging is an effective method of limb salvage after resection of a malignant or locally invasive benign bone tumor. Use of cancellous bone graft alone is less effective in achieving extracortical bone-bridging. The present study was performed to investigate the effects of a corticocancellous onlay graft on bone and soft-tissue formation over a porous-coated replacement prosthesis in the mid-diaphyseal region of canine femora. METHODS: Bilateral resection of a six-centimeter segment of the femoral diaphysis and reconstruction with a porous-coated segmental prosthesis was performed in six mongrel dogs. In one limb (the experimental side), eight strips of corticocancellous bone were evenly placed around the junctions between the femur and the prosthetic surface. Cancellous bone was placed under and between the strips of cortical bone. No graft was used in the other limb (the control side). The animals were followed for twelve weeks, with sequential assessments of load-bearing and radiographic evaluation. Biomechanical, histological, and microradiographic analyses of the specimens were performed after death. RESULTS: On the control side, load-bearing at four weeks postoperatively was significantly decreased compared with the preoperative value (p<0.05); no difference in these values could be detected on the experimental side. Both the area of the callus and the contact area between the bone and the prosthetic shoulder were greater on the experimental side (p<0.05). The mechanical stiffness and the maximum torque at failure of the extracortical bridging tissue across the junction between the bone and the prosthetic shoulder were eighteen (p<0.007) and five times greater (p<0.05), respectively, on the experimental side. CONCLUSIONS: Extracortical bone-bridging was accomplished with corticocancellous onlay bone-grafting. Without bone-grafting, bone formed only occasionally. Bone-grafting also enhanced the formation of a soft-tissue capsule around the prosthesis.

Animals↗

PubMed1999

Development of a new high-speed measuring system to analyze the dental cast form.

A new non-contact measurement system employing of a line laser and CCD camera was developed to rapidly measure maloccluded dental casts with large depth and high accuracy. In this system, the measuring interval along the line laser and the model scanning direction perpendicular to the line laser were both 0.2 mm. By averaging the raw data through repetitious input and smoothing by local filtering, the noise was effectively decreased. Sub-pixel processing, which calculates the pixel position with the largest brightness within one pixel, improved the accuracy in the measurement to 22 microns compared with 65 microns before sub-pixel processing. Dental casts with malocclusion were measured with this system. The time and the depth for the measurement were about 10 minutes and 60 mm, respectively. This was a vast improvement compared with our previous system employing a point laser. In the CG of the measured dental casts, smooth and detailed surface morphology were sufficiently reproduced, and recognition of the characteristic points in the cast could be accomplished with the accuracy of 30 microns. This measurement system will be helpful to analyze the dental cast form with malocclusion in progress.

Humans↗

PubMed1999

The serum interleukin 8 level reflects hepatic mitochondrial redox state in hyperthermochemohypoxic isolated liver perfusion with use of a venovenous bypass.

BACKGROUND: We have recently developed a simple method of hyperthermochemohypoxic isolated liver perfusion (HILP) as a regional therapy for unrecognized liver micrometastases. However, little is known about the influence of HILP on cytokine production and liver function. We investigated the influence of HILP on interleukin 8 (IL-8) production and the hepatic mitochondrial function and assessed the relationship between these 2 parameters. We also measured the serum tumor necrosis factor-alpha (TNF-alpha) and interleukin 1 beta (IL-1 beta) levels to examine the involvement of HILP-induced cytokines in the tumor response. METHODS: Sixteen patients with metastatic liver tumors were randomly assigned to undergo hepatectomy with HILP (group A, n = 9) or hepatectomy alone (group B, n = 7). The isolated liver was perfused for 30 minutes with Ringer's lactate solution containing chemotherapeutic agents warmed to 42 degrees C to 43 degrees C without oxygenation. RESULTS: The serum IL-8 levels in group A were markedly increased, with peaks at 3 hours after reperfusion, which was significantly higher than levels in group B (P < .01). In group A the arterial ketone body ratio, which reflects the hepatic mitochondrial redox state, decreased during perfusion and was gradually restored to the preperfusion level 1 hour after reperfusion. However, in group B it decreased during hepatectomy but rapidly recovered 5 minutes after hepatectomy. There was a significant negative correlation between the peak serum IL-8 level and the initial velocity of arterial ketone body ratio recovery for the first 5 minutes after reperfusion r = -0.83, P < .001). The serum TNF-alpha and IL-1 beta were temporarily detected only in 3 of 9 patients in group A. CONCLUSIONS: We have shown that HILP resulted in augmented IL-8 release but not TNF-alpha and IL-1 beta and that the serum IL-8 level reflects the hepatic mitochondrial redox state. These findings suggest that IL-8 production may be associated with hepatic mitochondrial impairment during ischemia. This work may contribute to new therapeutic strategies not only for hepatic ischemia reperfusion injury but also for metastatic liver tumors.

Aged↗

PubMed1999

The penetration characteristic of radio wave frequencies from oral cavities.

Recently many radio telemetry systems have been inserted into oral cavities and have been used to send data to receivers. Although various frequencies were used in those systems, there has been no report about the most effective penetrative frequency in the human body. In this study, we observed the penetration of radio waves from oral cavities and compared the efficiency of different frequencies. We selected 11 edentulous persons as subjects and the frequencies ranged from 1 to 150 MHz. Although the attenuation gradually increased with the increase of frequency in most cases, we observed that there were effective resonance-like frequencies in three subjects and the frequency differed from person to person. This phenomenon was only observed when receiving antennas were set beside the mouth and not observed when the antennas were set about 1 m away from subjects. Therefore, it is considered that this phenomenon was produced by the relationship between the transmitting antenna and the receiving antenna. This result suggested that there were effective resonance-like frequencies in the case of radiation from edentulous oral cavities.

Aged↗

PubMed1999

Erythropoietic, neurotrophic, and angiogenic functions of erythropoietin and regulation of erythropoietin production.

Stimulation of erythrocyte formation has been generally thought to be the sole physiological function of erythropoietin. Erythropoietin and its receptors, however, are also expressed in the central nervous system. In addition, erythropoietin is produced in the uterus in an estrogen-dependent manner. Novel functions of erythropoietin in these local sites are presented.

Central Nervous System↗

PubMed1998

Calcium is required for quasi-lipoxygenase activity of hemoproteins.

Bovine and guinea pig heart homogenates, porcine leukocyte homogenate, and human hemolysate were found to vigorously oxidize linoleic acid, with lipoxygenase-like activity, to its hydroperoxy, epoxy, hydroxy-epoxy, and keto compounds in the presence of calcium chloride. In the absence of calcium, the reaction was significantly reduced. Attempts to characterize this quasi-lipoxygenase activity revealed that calcium potentiated the quasi-lipoxygenase activities of hemoproteins (hemoglobin, myoglobin, myeloperoxidase, catalase, cytochrome c) and hemin at the physiological pH of 7.5. Lipid peroxidation by hemoproteins was inhibited by albumin and erythrocyte membranes in blood, as well as by a low concentration of calcium in cells. However, it seems possible that in extracellular fluid, which contains a high concentration of calcium and a low concentration of albumin, hemoprotein released from damaged cells could oxidize unsaturated fatty acids derived by phospholipase-A2 from phospholipids of damaged cellular membranes. In a model of quasi-lipoxygenase activation under such conditions, lipids of erythrocyte membranes were oxidized by hemoglobin in the presence of phospholipase-A2 and calcium. The effect of nitrogen oxide, paraquat, and bleomycin on oxidation by hemoproteins and hemin was also discussed.

Animals↗

PubMed1998

Estrogen-dependent production of erythropoietin in uterus and its implication in uterine angiogenesis.

Although erythropoietin (Epo) has been shown to possess in vitro angiogenic activity, its physiological significance has not been demonstrated. Normally angiogenesis does not occur actively in adults but an exception is the female reproductive organ. In the uterine endometrium, angiogenesis takes place actively for supporting the endometrial growth that occurs during transition from the diestrus to estrous stage. This transition is under control of 17beta-estradiol (E2), an ovarian hormone, and can be mimicked by injection of E2 to ovariectomized (OVX) mouse. Thus, the uterus is a pertinent site to examine the Epo function in angiogenesis. We found that Epo protein and its mRNA were produced in an E2-dependent manner, when the uterus from OVX mouse was cultured in vitro. The de novo protein synthesis was not needed for E2 induction of Epo mRNA. Administration of E2 to OVX mouse induced a rapid and transient increase in Epo mRNA in the uterus. Injection of Epo into the OVX mouse uterine cavity promoted blood vessel formation in the endometrium. Furthermore, injection of the soluble Epo receptor capable of binding with Epo into the uterine cavity of non-OVX mouse in diestrus stage inhibited the endometrial transition to proestrus stage, whereas heat-inactivated soluble Epo receptor allowed the transition to occur. These results, combined with our finding that the endothelial cells in uterine endometrium express Epo receptor, strongly suggest that Epo is an important factor for the E2-dependent cyclical angiogenesis in uterus.

Animals↗

PubMed1998

Involvement of protein kinase C and Src family tyrosine kinase in Galphaq/11-induced activation of c-Jun N-terminal kinase and p38 mitogen-activated protein kinase.

Mitogen-activated protein kinases (MAPKs) are activated by various extracellular stimuli. The signaling pathways from G protein-coupled receptors to extracellular signal-regulated kinase have been partially elucidated, whereas the mechanisms by which G protein-coupled receptors stimulate c-Jun N-terminal kinase (JNK) and p38 MAPK activities remain largely unknown. We have recently demonstrated that the signal from Gq/11-coupled m1 muscarinic acetylcholine receptor to p38 MAPK is mediated by both Galphaq/11 and Gbeta gamma in HEK-293 cells (Yamauchi, J., Nagao, M., Kaziro, Y., and Itoh, H. (1997) J. Biol. Chem. 272, 27771-27777). In the present study, we report that a constitutively activated mutant of Galpha11 (Galpha11 Q209L) activated not only p38 MAPK, but also JNK, and the activation of JNK and p38 MAPK by Galpha11 Q209L was partially inhibited by prolonged treatment with phorbol 12-myristate 13-acetate and calphostin C. In addition, the Galpha11 Q209L-stimulated activation of both kinases was blocked by a specific inhibitor of protein tyrosine kinases (PP2) and Csk (C-terminal Src kinase). Furthermore, we demonstrated that Galpha11 Q209L stimulated Src family kinase activity and induced tyrosine phosphorylation of several proteins in HEK-293 cells. These results suggest that Galphaq/11 stimulates JNK and p38 MAPK activities through protein kinase C- and Src family kinase-dependent signaling pathways.

Calcium-Calmodulin-Dependent Protein Kinases↗

PubMed1998

The delta isoform of protein phosphatase type 1 is localized in nucleolus and dephosphorylates nucleolar phosphoproteins.

The immunolocalization and substrates of protein phosphatases present in nucleolus were investigated using Swiss 3T3 cells and Novikoff hepatoma ascites cells. The protein phosphatase activity was detected in the extract of the isolated nucleoli and its activity was inhibited by okadaic acid with IC50 value of 160 nM. Immunoblotting assay indicated that PP1c delta but not PP1c alpha, PP1c gamma 1, and PP2Ac was localized in the isolated nucleoli. Confocal microscopy showed that PP1c delta was localized in nucleoli, nuclei, and cytosol, though the intensity of fluorescence at the nucleoli was stronger than that of the cytosol or nuclei. PP1c delta was co-localized with the major nucleolar phosphoprotein B23 at nucleoli. The phosphatase was capable of dephosphorylating several proteins in the nucleolus, including B23. The Km of PP1 for the recombinant B23.1, phosphorylated by endogenous kinase(s), was 3.5 microM. These results indicate that PP1c delta is the major serine/threonine phosphatase present in nucleolus and it dephosphorylates nucleolar phosphoproteins, including B23.

3T3 Cells↗

PubMed1998

Subcellular localization of protein phosphatase type 1 isotypes in mouse osteoblastic cells.

The cytolocalization of protein phosphatase type 1 catalytic subunits in exponentially growing mouse osteoblastic MC3T3-E1 cells was determined. Formaldehyde-fixed and alcohol-permeabilized cultured cells were reacted with the PP1 alpha, PP1 delta, PP1 gamma 1, and PP1 gamma 2 antibodies using immunohistochemical methods. With PP1 alpha antibody intense staining occurred in the nuclei, while with PP1 delta antibody nucleolus-like bodies were intensely stained. PP1 gamma 1 localized in the perinuclear region and in the nucleus of the cultured cells, with the staining reaction of the former being much stronger than that in the latter. An immunoreaction did not occur in the cells interacted with PP1 gamma 2 antibody or with the normal rabbit serum. Proteins were prepared from the exponentially growing cells and subconfluent cells. Cellular fractionation was also done with the exponentially growing cells and proteins were prepared from each fraction. Each protein preparation was subjected to SDS-PAGE followed by Western blot analysis with the antibodies. PP1 alpha recognized the 38 kDa proteins mainly present in the nucleus, whereas PP1 delta interacted with the proteins in the nucleolar fraction whose molecular weight was estimated as 37 kDa. PP1 gamma 1 antibody recognized a band corresponding to an estimated molecular weight of 36 kDa mainly in the cytosolic fraction. PP1 gamma 2 antibody and the normal rabbit serum did not interact with any proteins prepared from the cultured cells. Our observations show that four different isozymes of protein phosphatases occupy distinct compartments in MC3T3-E1 cells. This differential distribution suggests that these isozymes may play different roles in cellular functions.

Amino Acid Sequence↗

PubMed1998

Genetic determinant and environmental carcinogens.

2-Amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) is the most abundant mutagenic heterocyclic amine (HCA) present in cooked foods. PhIP induces colon cancer in male Fischer 344 (F344) rats, and its role in human colon carcinogenesis has been suspected. To study the ecogenetics in PhIP colon carcinogenesis, rat system using aberrant crypt focus (ACF) formation as a phenotypic marker was applied. Among Buffalo (BUF), Brown Norway (BN), F344 and ACI/N (ACI) strains of rats, F344 rats produced a lower level of PhIP-DNA adducts than other three strains, and the number of ACFs/rat was highest in BUF, intermediate in BN and F344 and lowest in ACI. Thus there was no correlation between adduct levels and number of ACF induced by PhIP. F1 progenies of BUF and ACI developed ACF at a similar level to that of F344, and F1 progenies of F344 and ACI developed ACF at a similar level to that of F344. Thus it was indicated that susceptibility of F344 to the ACF induction was autosomally dominant over ACI rats. The results also suggest that BUF rats have at least two genes, one is autosomally recessive against ACI rats and one is autosomally dominant similar to that F344 has. A total of 170 progeny of ACI backcross of F344/ACI F1 were examined for number of ACFs and 65 progeny were phenotyped as F344 and 60 were ACI. Using these 125 rats, chromosomal mapping is being performed using markers of simple sequence length polymorphism (SSLP) and representational difference analysis (RDA). By mapping the gene, we will be able to identify humans who might belong to high risk group in general population, and cancer can be prevented more efficiently by attaining early diagnosis.

Animals↗

PubMed1998

Evidence for an antemortem injury of a burned head dissected from a burned body.

A 41-year-old woman was killed following blows to the head with a bat. Her body was burned and her head was dissected with a saw and hidden in the soil. Although the body was discovered within a few days of her death, the putrescent head was not located until four months later. A fracture of the left temporal bone was found in the head, which was partially charred. The confession of the murder suspect was unreliable, and the police suggested that the cause of death may not have been a head injury but asphyxia, and that the bone fracture to the head may have been caused after its dissection. It was therefore necessary to determine when the fracture was formed. The fracture line continued to the base of the skull, indicating that it was not due to heat. Magnetic resonance computed tomography (MR-CT) indicated the existence of a blood clot in the left mastoid cells, across which the fracture line passed. Upon sectioning of the skull with a saw, a dark red clot was located in the left mastoid cells, indicating that bleeding had occurred before the body was set alight. It seemed most logical to assume that the bone fracture was formed when the victim was alive, or before her body was burned.

Adult↗

PubMed1998

No direct correlation between mutant frequencies and cancer incidence induced by MeIQ in various organs of Big Blue mice.

The carcinogenicity of 2-amino-3,4-dimethylimidazo[4,5-f]quinoline (MeIQ) was examined in Big Blue female mice with the genetic background of C57BL/6N. With the administration of 300 ppm of MeIQ in their diet for 92 weeks, the Big Blue female mice developed intestinal tumors and hepatocellular carcinomas. The incidences of adenocarcinomas were 42% (8/19) in the colon and 68% (13/19) in the cecum. The incidence of hepatocellular carcinomas was 84% (16/19). No carcinomas of the intestine or the liver were induced in the control group. As we previously reported, administration of 300 ppm of MeIQ in a diet for 12 weeks induced lacI mutants at the highest frequency in colonocytes, and at only less than one-tenth of the colon in cells of the liver, forestomach and bone marrow, indicating no direct correlation between the lacI mutant frequency (MF) and cancer incidence (CI). The fate of cells with lacI mutation in each organ should be taken into consideration to validate MF as an indicator of carcinogenic potency of a chemical in different organs.

Animals↗

PubMed1998

In vivo evidence that erythropoietin protects neurons from ischemic damage.

Erythropoietin (EPO) produced by the kidney and the liver (in fetuses) stimulates erythropoiesis. In the central nervous system, neurons express EPO receptor (EPOR) and astrocytes produce EPO. EPO has been shown to protect primary cultured neurons from N-methyl-D-aspartate (NMDA) receptor-mediated glutamate toxicity. Here we report in vivo evidence that EPO protects neurons against ischemia-induced cell death. Infusion of EPO into the lateral ventricles of gerbils prevented ischemia-induced learning disability and rescued hippocampal CA1 neurons from lethal ischemic damage. The neuroprotective action of exogenous EPO was also confirmed by counting synapses in the hippocampal CA1 region. Infusion of soluble EPOR (an extracellular domain capable of binding with the ligand) into animals given a mild ischemic treatment that did not produce neuronal damage, caused neuronal degeneration and impaired learning ability, whereas infusion of the heat-denatured soluble EPOR was not detrimental, demonstrating that the endogenous brain EPO is crucial for neuronal survival. The presence of EPO in neuron cultures did not repress a NMDA receptor-mediated increase in intracellular Ca2+, but rescued the neurons from NO-induced death. Taken together EPO may exert its neuroprotective effect by reducing the NO-mediated formation of free radicals or antagonizing their toxicity.

Animals↗

PubMed1998

High frequency of beta-catenin (ctnnb1) mutations in the colon tumors induced by two heterocyclic amines in the F344 rat.

Activating mutations in the beta-catenin (CTNNB1) gene corresponding to N-terminal phosphorylation sites in the protein have been implicated in the development of human colon cancer. To determine the possible involvement of such mutations during chemically induced colon carcinogenesis, we examined the corresponding region of Ctnnb1 in colon tumors induced in the F344 rat by two cooked meat heterocyclic amines, 2-amino-3-methylimidazo[4,5-f]quinoline and 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP). All of the colon tumors induced by 2-amino-3-methylimidazo[4,5-f]quinoline that were examined (5 of 5) and 4 of 7 PhIP-induced colon tumors had mutations within or flanking codons corresponding to important phosphorylation sites in beta-catenin. None of the colon tumors bearing Ctnnb1 mutations had genetic changes in the Apc gene, and those that contained wild-type Ctnnb1 were known from our previous work to contain Apc mutations. The results provide evidence for a major role of the beta-catenin/Apc pathway in the development of heterocyclic amine-induced colon tumors and give further weight to the view that regulation of beta-catenin is critical to the tumor suppressive effects of Apc during colon carcinogenesis. In contrast, Ctnnb1 mutations were completely absent in 23 PhIP-induced mammary tumors, in accordance with recent work showing that human breast carcinomas lack mutations in CTNNB1.

Animals↗

PubMed1998

NO is not involved in the simvastatin induced cell division and differentiation in PC12 cells.

Simvastatin, a potent 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase inhibitor has been reported to inhibit cell division and induce neurite-like outgrowth in PC12 cells [Sato-Suzuki, I. and Murota, S., Neurosci. Lett., 220 (1996) 21-24]. In the present paper, we examined whether the induced nitric oxide (NO) in the simvastatin-treated PC12 cells is involved in the growth arrest and differentiation as reported in nerve growth factor (NGF) treated PC12 cells. Treatment of PC12 cells with simvastatin caused peripherin formation and enhanced NO production just like NGF-treated PC12 cells. Different from NGF, however, NO synthase inhibitors could not affect the growth arrest and differentiation in simvastatin-treated PC12 cells. In conclusion, NO had nothing to do with cell division and differentiation in simvastatin-treated PC12 cells.

Animals↗

PubMed1998

Embryonal carcinoma P19 cells produce erythropoietin constitutively but express lactate dehydrogenase in an oxygen-dependent manner.

Embryonic stem cells and embryonal carcinoma P19 cells produce erythropoietin (Epo) in an oxygen-independent manner, although lactate dehydrogenase A (LDHA) is hypoxia-inducible. To explore this paradox, we studied the operation of cis-acting sequences from these genes in P19 and Hep3B cells. The Epo gene promoter and 3' enhancer from P19 cells conveyed hypoxia-inducible responses in Hep3B cells but not in P19 cells. Together with DNA sequencing and the normal transcription start site of P19 Epo gene, this excluded the possibility that the noninducibility of Epo gene in P19 cells was due to mutation in these sequences or unusual initiation of transcription. In contrast, reporter constructs containing LDHA enhancer and promoter were hypoxia inducible in P19 and Hep3B cells, and mutation of a hypoxia- inducible factor 1 (HIF-1) binding site abolished the hypoxic inducibility in both cells, indicating that HIF-1 activation operates normally in P19 cells. Neither forced expression of hepatocyte nuclear factor 4 in P19 cells nor deletion of its binding site from the Epo enhancer was effective in restoring Epo enhancer function. P19 cells may lack an unidentified regulator(s) required for interaction of the Epo enhancer with Epo and LDHA promoters.

Base Sequence↗

PubMed1998

Calcium ions potentiate lipoxygenase activity of cytochrome c at the physiological pH.

Cytochrome c oxidized linoleic acid, with lipoxygenase-like activity, to 9-hydroperoxy-10,12-octadecadienoic acid (9-HpODE), 13-hydroperoxy-9,11-octadecadienoic acid (13-HpODE), 13-keto-9,11-octadecadienoic acid (13-KODE), and hydroxy-epoxy-octadecenoic acids (HEPOs) in the presence of calcium chloride at the physiological pH of 7.5. In the absence of calcium, the reaction was remarkably reduced. Statistically significant increase of 13-KODE was observed 1 min after the reaction was started by adding calcium. Calcium chloride at concentrations as low as 30 microM caused statistically significant increase of 13-KODE production in the presence of 1 microM cytochrome c, and cytochrome c at concentrations as low as 10 nM caused statistically significant increase of 13-KODE production in the presence of 2 mM calcium. The addition of bleomycin enhanced the reaction, and the addition of adriamycin attenuated the reaction. The same oxidized linoleic acid products were found in the submitochondrial particles which were hydrolyzed by phospholipase-A2 in the presence of calcium. Lipoxygenase activity potentiated by calcium may be involved in the lipid peroxidation in the submitochondrial particles hydrolyzed by phospholipase-A2.

Animals↗