Search PubMed⌕ Search

Biomedical subjects

N Kaneda

Publications and source records attributed to N Kaneda.

At least 73 records · Page 4Linked to original sources
PubMed1991

Role of phagocytes in antimicrobial defence of the middle ear.

The role of phagocytes in the antimicrobial defence of the middle ear was investigated in this experiment, using Hartley strain guinea pigs with an experimental otitis media. Otitis media was induced with an inoculation of Streptococcus pneumoniae into the tympanic cavity through the ear drum. For depletion of peripheral blood phagocyte population such as monocytes and polymorphonuclear neutrophils (PMNs), whole body irradiation (250 rad or 500 rad) was carried out on guinea pigs three days before S. pneumoniae inoculation into the middle ears. Carrageenan was also used for selective depletion of mononuclear cells, to distinguish their role from polymorphonuclear neutrophils. In control animals, otitis media was induced reproducibly with middle ear inoculation of more than 10(6) S. pneumoniae. In irradiated animals, which underwent 10(2) or 10(4) S. pneumoniae inoculation, the incidence of otitis media because of S. pneumoniae infection became higher in accordance with the dosage of irradiation. However, no significant difference was seen in the occurrence of otitis media and the number of viable bacteria recovered from bulla washings between controls and carrageenan-treated animals. These results suggest that phagocytes, particularly neutrophils, are essential for antimicrobial defense at the early phase of the middle ear infection with S. pneumoniae.

Animals↗
PubMed1991

Congenital middle ear cholesteatoma: report of 3 cases.

In this report, we presented 3 cases of congenital middle ear cholesteatoma which occurred in a 12-year-old girl, a 4-year-old boy, and a 6-year-old boy. In all 3 cases, there was a whitish mass behind a normal tympanic membrane. Congenital middle ear cholesteatoma is not a rare disease. In the early stage, it is asymptomatic. But when it progresses, this disorder can destroy conductive systems of the middle ear and cause many symptoms. One patient (Case 1) had a complaint of hearing impairment. She underwent mastoidectomy and tympanoplasty; however, the cholesteatoma recurred. The other 2 patients had no symptoms. The abnormal appearance of their tympanic membrane was found by chance at their local otologists. We performed tympanotomies and removed cholesteatomas without aftereffects. When otologists note an abnormal appearance behind a normal tympanic membrane, with or without symptoms, tympanotomy should be done due to the possibility of congenital middle ear cholesteatoma.

Audiometry↗
PubMed1991

CPT-11 converting enzyme from rat serum: purification and some properties.

A rat serum enzyme that catalyzes the conversion of a pro-drug, 7-ethyl-10-[4-(1-piperidino)-1-piperidino] carbonyloxycamptothecin (CPT-11), to an anticancer drug, 7-ethyl-10-hydroxycamptothecin (SN-38), was purified and its properties were characterized. The enzyme was purified by column chromatography on diethylaminoethyl Toyopearl 650M, QAE-Sephadex, Sephadex G-150, Con A-Sepharose and high performance liquid chromatography with an ion-exchanger column. It was most active at pH 7.5 and was stable at pH 4-9 for 1 h at 30 degrees C. The molecular weight was estimated to be 60 and 57 kDa by gel filtration and sodium dodecylsulfate-polyacrylamide gel electrophoresis methods, respectively, and the isoelectric point was 4.6, as determined by isoelectric focusing. The Km value for CPT-11 was 0.28 microM. This enzyme was inhibited by diisopropyl phosphorofluoridate (DFP) and phenylmethanesulfonyl fluoride (PMSF) but insensitive to eserine, p-chloromercuribenzoate (PCMB) and ethylenediaminetetraacetate (EDTA). The enzyme also hydrolyzed p-nitrophenylacetate (p-NPA), a commonly used substrate for esterases, but was not active toward acetylcholine, suggesting that the enzyme is a carboxylesterase[EC 3.1.1.1]. During the hydrolyses of CPT-11 and p-NPA, an initial burst phenomenon similar to that found in the alpha-chymotrypsin-catalyzed hydrolysis of p-NPA was observed. Kinetic analysis revealed that the deacylation of the enzyme is the rate-limiting step in substrate hydrolysis. This enzyme was found to also split other ester derivatives of SN-38 besides CPT-11.

Animals↗
PubMed1990

Right-left asymmetry of tyrosine hydroxylase in rat median eminence: influence of arterial baroreflex nerves.

Arterial baroreceptor nerves consist of 2 pairs of nerves, carotid sinus and aortic. The latter nerves differ bilaterally as to size and origin suggesting that central baroreceptor activity could be asymmetrical. This was tested by studying activity of tyrosine hydroxylase (TH), the rate limiting enzyme for catecholamine synthesis, on right and left sides of the brain in median eminence (ME), arcuate nucleus (ARCN), paraventricular nucleus (PVN) and supraoptic nucleus (SON). Seven days after right or left sinoaortic denervation (SAD), bilateral SAD or sham-operation (SO), in situ TH activity was determined by the rate of dihydroxyphenylalanine (DOPA) accumulation following administration of a dopa decarboxylase inhibitor. Unexpectedly, the intact, SO rats had significantly lower DOPA concentrations on the left than right sides of ME in two separate studies; right-left differences averaged 24% and 38%. Evidence supporting asymmetry of TH activity was the lower concentrations of catecholamines on left than right sides of the brain in intact SO rats, namely, dopamine in ME and ARCN and norepinephrine and epinephrine in ARCN. Unilateral and bilateral SAD further suppressed TH activity on the left side of the ME; the maximum right-left difference was 58% in rats with right SAD. Their ARCN also showed asymmetry of TH activity. Asymmetry of TH activity was attributable to asymmetry of TH protein which was significantly lower on the left sides of ME and ARCN. This is the first demonstration of bilateral asymmetry of TH in mediobasal hypothalamus and its enhancement by interruption of left or right afferents from the arterial vascular system.

Animals↗
PubMed1990

Metabolism and pharmacokinetics of the camptothecin analogue CPT-11 in the mouse.

A new water-soluble derivative of camptothecin, 7-ethyl-10-[4-(1-piperidino)-1-piperidino]carbonyloxycamptothecin (CPT-11), did not exhibit potent antitumor activity in vitro against experimental tumor cells. The 50% effective doses of CPT-11 against KB and L1210 cells were 1100 and 5500 ng/ml, respectively. These values were markedly higher than those of camptothecin (CPT, 0.98 and 3.7 ng/ml) or 7-ethyl-10-hydroxycamptothecin (SN-38, 0.37 and 3.6 ng/ml). CPT-11 was found to be converted into SN-38 in mouse serum. In vitro incubation of CPT-11 in mouse serum or tissue homogenate enhanced the growth-inhibitory activity much more than that expected from the concentration of CPT-11. This enhancement of the activity coincided with that expected from the SN-38 concentration in incubated serum or homogenate, though the contribution of CPT-11 could not be refuted. SN-38 is considered to play a major role in the antitumor activity when CPT-11 is incubated in serum or homogenate. The plasma CPT-11 concentration decreased biexponentially after i.v. administration of CPT-11 into mice with a biological half-life of 0.8 to 1.1 h. The area under the plasma CPT-11 concentration-time curve showed dose dependency. The SN-38 concentration decreased for the first 30 min after administration and was then maintained for a few hours at about 0.1 microgram/ml after i.v. administration of 20 and 40 mg/kg of CPT-11 followed by the log-linear terminal phase with a half-life of about 2 h which was independent of the dose. It is suggested that the maintenance of plasma SN-38 concentration might be necessary for it to exhibit antitumor activity in vivo.

Animals↗
PubMed1990

Nonlinear pharmacokinetics of CPT-11 in rats.

The pharmacokinetics of a new water-soluble derivative of camptothecin. 7-ethyl-10-[4-(1-piperidino)-1-piperidino]carbonyloxycamptothecin (CPT-11), and its major metabolite, 7-ethyl-10-hydroxycamptothecin (SN-38), was investigated after i.v. administration of 1 to 40 mg/kg of CPT-11 to rats. The plasma concentration of CPT-11 decreased biexponentially. The area under the concentration-time curve increased nonlinearly as the dose increased. SN-38 was found in the plasma, bile, urine, and feces. The SN-38 level was maintained at 0.06 to 0.08 micrograms/ml for 0.5 to 5.5 h depending on the dose, followed by exponential decay. Thirty-three to 58% of the CPT-11 was excreted without metabolism into the bile and urine for 24 h. SN-38 was mainly excreted into the bile. Analysis of the clearance has shown nonlinear pharmacokinetics which was due to metabolic processes such as the conversion of CPT-11 to SN-38.

Animals↗
PubMed1990

Tetrahydrobiopterin-dependent production of L-dopa in NRK fibroblasts transfected with tyrosine hydroxylase cDNA: future use for intracerebral grafting.

In the present study, tyrosine hydroxylase (TH; EC 1.14.16.2) cDNA was transfected into cultured fibroblasts and the production of L-3,4-dihydroxyphenylalanine (L-DOPA) was determined. Type 2 TH cDNA was transfected into fibroblasts (NRK-49F) derived from the normal rat kidney, and the expression of the TH minigene was screened by immunocytochemical staining and immunoblotting analysis with TH antiserum. Several clones of the NRK transfectants that produce TH molecules were obtained. The expressed TH molecules showed high enzyme activity in a complete assay system in vitro. However, L-DOPA was not detected in the cultured cells due to the possible absence of de novo synthesis of (6R)-L-erythro-tetrahydrobiopterin (BH4) in these cells. When BH4 was added to the medium, a large amount of L-DOPA was detected not only in the cells but also in the medium. These findings may aid in regulating the amount of L-DOPA secretion from cells after they are transplanted into the brain.

Animals↗
PubMed1990

Optimal conditions for protease use in the assay of serum mitochondrial aspartate aminotransferase.

The optimal conditions for selective proteolytic inactivation of cytosolic aspartate aminotransferase (c-AST) to determine mitochondrial aspartate aminotransferase (m-AST) in serum were studied. Protease 401 was found to be effective over a pH range of 6.0-10.0. A pH of 9.5 with 0.5% albumin in the reagent mixture was determined to be optimal for inactivation of c-AST and preservation of m-AST, lactic dehydrogenase (LDH), and malic dehydrogenase (MDH) in the assay procedure. The presence of serum endogenous protein inhibitors such as alpha 1-antitrypsin and alpha 2-macroglobin did not inhibit protease 401.

Aspartate Aminotransferases↗
PubMed1990

Neuroleptic-induced vacuous chewing movements as an animal model of tardive dyskinesia: a study in three rat strains.

Vacuous chewing movements (VCMs) in three different rat strains developed at considerably different rates after 19 weeks of continual haloperidol treatment at an average daily dose of 1.5 mg/kg. Sprague Dawley (SD) rats displayed relatively high rates of VCMs with low variability, compared to Wistar (W) and Long Evan (LE) rats. Atropine decreased but did not abolish VCMs in two of the three strains (LE greater than SD). After haloperidol withdrawal, VCMs remitted gradually in all strains, but least rapidly in the SD rats. In a separate group of SD rats. VCMs were rated weekly from the start of haloperidol treatment and showed considerable interindividual variability. Even after 24 weeks of continuous haloperidol, 12 out of 32 treated rats showed no VCMs at all, while 13 out of 32 had intense movements, analogous to the clinical situation in which only some patients treated with neuroleptics develop tardive dyskinesia. These results indicate that there are individual and strain differences in the development of VCMs, and suggest that there may also be genetically determined differences in the development of tardive dyskinesia.

Animals↗
PubMed1990

Comparative studies on the usefulness of phosphate versus glycerin enema in preparation for colon examinations.

Fifty of 100 persons who had undergone health screening received phosphate enema while the other 50 received glycerin enema prior to proctoscopy and barium enema, and their usefulness for preparation for colon examination was compared by a double-blind test. There was no significant difference in the degree of colonic cleansing achieved by proctoscopy and barium enema. In the subjects who received phosphate enema, the incidence of abdominal pain was less than that in those who received glycerin enema, while the effect of phosphate enema on defecation appeared later than that of glycerin enema, indicating prolonged stool retention in the subjects given phosphate enema. To study the safety of the two enemas, either phosphate enema, glycerin enema or physiological saline solution as a control was administered at 0.35 ml/animal in the rectum by 4-h closure of the anus in 10 male 7-week-old Wistar rats, and the rectal mucosa was observed for irritation macroscopically and histopathologically. Glycerin enema produced less irritation than phosphate enema diffusely in the entire area of the rectum, while phosphate enema produced more local irritation at the end of the rectum than glycerin enema. The differences in the extent of irritation and injury between phosphate and glycerin enemas were considered to be derived from differences in the pharmacologic actions of these drugs. If the extent of injury were included in the extent of irritation, the difference in irritation between phosphate and glycerin enemas would not be significant. As described above, no specific difference seem to exist in the usefulness of phosphate and glycerin enemas as preparation for colon examination.

Animals↗
PubMed1990

Proteolytic measurement of mitochondrial aspartate aminotransferase in human serum.

A new proteolytic measurement of serum mitochondrial aspartate aminotransferase was evaluated using cytosolic aspartate aminotransferase inactivating protease. Some of the proteases, such as, alpha-chymotrypsin, subtilisin and cytosolic aspartate aminotransferase inactivating protease 401 from Streptomyces species, also specifically inactivated cytosolic aspartate aminotransferase, but not mitochondrial, aspartate aminotransferase. The protease 401 was the most heat stable for storage and showed a higher inactivation rate for cytosolic aspartate aminotransferase--up to 7000 IU/L--more than 200-fold the upper limit. The coefficient of variation of the proteolytic method was less than 10%. Results by the present method correlated with those by the immunochemical method (r = 0.970) and the regression curve was Y = 0.95X + 1.60 (Y: immunochemical method; X: proteolytic method). In the present assay system, reference values for mitochondrial aspartate aminotransferase activity in 500 healthy people ranged from 2.0-7.2 U/L (mean 3.8 U/L).

Aspartate Aminotransferases↗
PubMed1990

[A case of metastatic malignant melanoma of the stomach treated by a surgical operation].

A metastatic malignant melanoma of the stomach is not a disease of rate occurrence, but reports of melanomas that were clinically diagnosed and were treated by a surgical operation are rare. The case presented is a 73-year-old woman, who underwent an excision of a malignant melanoma of the left sole in 1981. Subsequently, she had a subcutaneous metastasis of a melanoma three times and was treated by a resection and immunochemotherapy. In January, 1988, an X-ray examination of stomach revealed a giant elevated lesion, and an endoscopic examination of the stomach revealed a black pigmented tumor with ulceration. A biopsy taken from the tumor confirmed the diagnosis of a malignant melanoma. Thus, she was given a total gastrectomy. Reports in the Japanese literature of metastatic malignant melanomas of the stomach diagnosed while the patient was still living amount to 11 cases. Of this number, 3 patients were given a surgical operation. Further, these 3 patients lived longer than the non-surgically treated cases and had a better quality of remaining life.

Aged↗
PubMed1989

Synthesis of L-3,4-dihydroxyphenylalanine by tyrosine hydroxylase cDNA-transfected C6 cells: application for intracerebral grafting.

In the present study, we obtained genetically manipulated nonneuronal cells which synthesize a catecholamine precursor for future use in intracerebral grafting. Human type 1 tyrosine hydroxylase (TH; EC 1.14.16.2) cDNA was inserted into eukaryotic expression vector pKCRH2 and was co-transfected into C6 cells with plasmid pSV2neo. Expression of the TH minigene was screened by immunohistochemical staining with TH antibody and immunoblotting analysis. Several clones of the C6 transfectants that produce TH molecules were obtained. These cells showed TH activity, and the product, L-3,4-dihydroxyphenylalanine (L-DOPA), was detected intracellularly due to the absence of L-amino acid decarboxylase (EC 4.1.1.28) activity. It was found that a large amount of L-DOPA was released from the cells into the culture medium. These transfectants were transplanted into rat brain, and the expression of TH was examined immunohistochemically. On the 10th day following transplantation, a mass of C6 cells which was heavily stained with TH antibody was observed in the brain. These findings may provide us with an opportunity to investigate the effects of intracerebral transplantation of nonneuronal cells that produce catecholamine or its precursor.

Animals↗
PubMed1989

[Clinical evaluation of a new hand-held impedance audiometer].

In order to evaluate the accuracy and clinical availability of MicroTymp (Welch Allyn) which was new, portable, cordless, and hand-held impedance audiometer, 64 normal subjects and 52 patients with otitis media with effusion were examined by use of MicroTymp and Amplaid 702 (Dana Japan). The types of tympanogram were agreed with each other in 87% of ears. Six cases of type C tympanograms in MicroTymp were shown to be type A by Amplaid 702, and the average of peak pressure of MicroTymp was more negative than Amplaid 702. The difference of peak pressure seemed to be influenced by the speed of ear canal pressure change, because the speed of MicroTymp was considerably higher than Amplaid 702; -200mmH2O/sec in MicroTymp, and -50mmH2O/sec in Amplaid 702. In the experimental study on 7 normal ears, negative shift of peak pressure was observed by increasing the rate of ear canal pressure change from -25mmH2O/sec to -50mmH2O/sec in Amplaid 702. These findings suggest that one should note in the judgment of type C tympanogram in MicroTymp having the peak pressure close to -100mmH2O.

Acoustic Impedance Tests↗
PubMed1989

[Pathogenesis of Parkinson's disease, a molecular genetic approach].

In juvenile parkinsonism (JP), unlike naturally occurring Parkinson's disease, high frequency of familial onset is observed, which suggests the involvement of some genetic factor(s) in the pathogenesis of the disease. In an attempt to conduct a molecular genetic approach to JP, we tried to isolate tyrosine hydroxylase (TH) cDNA from human pheochromocytoma, and demonstrated the existence of four types of cDNA (type 1, 2, 3 and 4), differing in the 5'-terminal region. All four cDNAs had the same sequence in common from ATG of the translation start codon to 90th nucleotide. However, in types 2, 3 and 4, characteristic sequences were inserted between 90th and 91 st nucleotides of type 1 cDNA. TH genomic DNA cloning showed that the multiple form of mRNA were produced from a single gene through alternative splicing. Four types of cDNA was expressed in COS cells. They exhibited different homospecific activities: type 1 TH having the highest activity, others less than 40% of type 1 TH. The question whether possible change in TH gene is related to the pathogenesis of JP is now being pursued based on these molecular biological understanding of TH gene.

Amino Acid Sequence↗