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Biomedical subjects

M Inagaki

Publications and source records attributed to M Inagaki.

At least 343 records · Page 19Linked to original sources
PubMed1988

A fourth type of rabbit protein kinase C.

Three rabbit cDNA clones coding for three types of protein kinase C (PKC alpha, beta, and gamma) have recently been identified and the structures determined [Ohno, S., Kawasaki, H., Imajoh, S., Suzuki, K., Inagaki, M., Yokokura, H., Sakoh, T., & Hidaka, H. (1987) Nature (London) 325, 161-166]. By use of these cloned cDNAs as hybridization probes, a fourth type (delta) of cDNA clone, which encodes a protein highly homologous to PKC alpha, beta, and gamma, was identified. PKC delta is composed of 697 amino acid residues and contains several peptide sequences determined at the protein level with the brain PKC preparation. This indicates that this molecular type (PKC delta) is, along with PKC alpha, beta, and gamma, a constituent of the brain PKC preparation. Sequence comparison among the four PKC types revealed that PKC delta is somewhat distinct from the other PKC types. PKC delta shows 99% amino acid sequence identity with rat PKC type I [Knopf, J. L., Lee, M.-H., Sultzman, L. A., Kriz, R. W., Loomis, C. R., Hewick, R. M., & Bell, R. M. (1986) Cell (Cambridge, Mass.) 46, 491-502], indicating relationship of these PKC types. The mRNA for PKC delta is exclusively concentrated in the brain.

Amino Acid Sequence↗
PubMed1988

Direct inhibitory projection of pontine omnipause neurons to burst neurons in the Forel's field H controlling vertical eye movement-related motoneurons in the cat.

This study examines the nature of the efferent projection of omnipause neurons (OPNs) in the midline pontine tegmentum to medium-lead burst neurons (BNs) in the Forel's field H (FFH), both of which exhibit activities related to vertical eye movements, using chronically prepared alert cats. Antidromic spikes of the BNs evoked by oculomotor nucleus stimulation were suppressed by shortly preceding (less than 5 ms) microstimulation within the OPN area including actual recording sites of OPNs. Many OPNs were antidromically activated by microstimulation at recording sites of the BNs. Furthermore, systematic tracking in and around the FFH with the stimulating microelectrode substantiated that the OPNs issued axonal branches within the BN area. These results suggest direct inhibitory projection of OPNs to the BNs.

Action Potentials↗
PubMed1988

The effect of an orally administered proteolytic enzyme on the elasticity and viscosity of nasal mucus.

We have evaluated the effect of serratiopeptidase (SER), a proteolytic enzyme, on the elasticity and viscosity of the nasal mucus in adult patients with chronic sinusitis. SER was administered in a dose of 30 mg/day orally for 4 weeks. Nasal mucus was collected from the nasal cavities of each patient before (week 0) and 4 weeks after the start of the medication (week 4). The storage modulus (G') and the dynamic viscosity (eta') of each specimen of nasal mucus were determined by an oscillating sphere magnetic rheometer at frequencies of 0.5, 1, 5, 10 and 20 Hz at a constant temperature of 25 degrees C. The dynamic viscosity (eta') of the mucus at week 4 was significantly lower than that at week 0 (at frequencies of 5, 10 and 20 Hz). No significant differences were observed in the storage modulus (G') between the mucus at week 0 and week 4. SER reduced the viscosity but not the elasticity of the nasal mucus. These findings are discussed in relation to mucociliary clearance.

Administration, Oral↗
PubMed1988

Rapid phosphorylation of MAP-2-related cytoplasmic and nuclear Mr 300,000 protein by serine kinases after growth stimulation in quiescent cells.

Antibody against brain microtubule-associated protein 2 (MAP-2) immunoprecipitated Mr 300,000 and 80,000 proteins of cultured fibroblasts and kidney cells. These proteins were not appreciably phosphorylated in quiescent cells, but were rapidly phosphorylated after growth stimulation by insulin, epidermal and fibroblast growth factors, transferrin, phorbol ester and diacylglycerol in the presence of Ca2+, in a manner similar to that of MAP-1-related Mr 350,000 protein (J. Cell Biol. 100, 748-753). A Ca2+ ionophore, which is known to make the quiescent cell competent but not to enter into the growth cycle, did not induce the phosphorylation. In a chase experiment, decay half lives of labeled phosphoproteins were 5 h for Mr 350,000 and 300,000 proteins, and 1.5 h for Mr 80,000 protein. On subcellular fractionation, phosphorylated Mr 350,000 and 300,000 proteins were detected first mainly in the cytoplasm and then in the nucleus, while Mr 80,000 phosphoprotein was consistently detected in the cytoplasm. The phosphorylation of these proteins occurred on serine residues after stimulation with various factors. Thus, the phosphorylation of cytoskeleton-associated Mr 350,000 and 300,000 proteins by serine kinases seems to be a common second process after growth stimulation and to link cytoplasmic and intranuclear events.

Animals↗
PubMed1988

The antibiotic susceptibilities and beta-lactamase production of clinical isolated Branhamella catarrhalis from acute otitis media in children.

Branhamella catarrhalis has been misconsidered as a normal resident in human respiratory tract for a long time. However, many authors recently have reported its pathogenecity and isolated it from the otolaryngological region. In our study, this organism can be isolated from the ear and nasal discharge in the child with acute otitis media by the rate of 7.5% and 21.4% respectively. Out of this 107 isolated strains, 97 strains (90.7%) were found to be beta-lactamase producing organisms. The MIC measurement of penicillins and cephems (except CEX) for inhibition of all these strains in our study is 6.25 micrograms/ml or less and because of the unreliability of the ABPC's susceptibility test by disk method, it is necessary to check the beta-lactamase production in each strain. Becoming of the high emergence rate of beta-lactamase producing strains, B. catarrhalis should be considered to be as important pathogen as Streptococcus pneumoniae and Haemophilus influenzae in upper respiratory tract infections in children.

Acute Disease↗
PubMed1988

Endotoxin-specific chromogenic assay for plasma in pregnant women, umbilical cords, neonates and children.

The concentration of endotoxin (Et) was measured by means of a new endotoxin-specific assay (Endospecy) in 15 paired samples of maternal and umbilical cord (UC) blood, and 38 neonates and 5 children. The Et levels in maternal and UC blood with normal pregnancy and delivery were 0.8 +/- 0.4 and 1.3 +/- 0.6 pg/ml, respectively. In some cases of premature rupture of the membrane, Et in maternal and UC blood was slightly increased. The level in non-infectious neonates was 6.7 +/- 5.1 pg/ml and that in children less than 10 pg/ml. However, patients with infections showed a high Et level, which decreased on antibiotic treatment. In addition, the pattern of the changes in Et in cerebrospinal fluid (CSF) did not parallel that in plasma. Therefore, in perinatal high-risk babies, the probability of affection with endotoxemia should be considered, but the effects of endotoxin on the central nervous system are presumably indirect because of the low permeability of Et through the blood-CSF-barrier.

Adult↗
PubMed1988

Atypical form of Menkes kinky hair disease with mitochondrial NADH-CoQ reductase deficiency.

A male infant with an atypical form of Menkes kinky hair disease showed mitochondrial NADH-CoQ reductase (complex I) deficiency in a femoris muscle biopsy. His clinical features consisted of hypotonicity of the upper limbs, hyper-reflexia of the lower extremities, abnormal hair and fine myoclonic movement of the hands. The serum levels of copper and ceruloplasmin were just below normal range, and the copper concentration in fibroblastic cells was much increased (101.2 ng/mg of protein). The occurrence of this case suggests that there may be a mild form of Menkes disease with a NADH-CoQ reductase deficiency or other mitochondrial enzyme defects.

Atrophy↗
PubMed1988

A fluorescent protein kinase C inhibitor: 1-(1-hydroxy-5-isoquinolinylsulfonyl)piperazine.

The isoquinolinesulfonamide compound 1-(5-isoquinolinyl-sulfonyl)-2-methylpiperazine (H-7) has been widely used as a protein kinase C inhibitor. Although H-7 and its derivatives are useful for the demonstration of the biological function of protein kinase C or cyclic nucleotide-dependent protein kinases, these compounds are not available for a histological approach to protein kinase C. In the present study, we introduce 1-(1-hydroxy-5-isoquinolinylsulfonyl)piperazine (hydroxy H-7) as a useful tool in tissue experiments. The property of the compound as a protein kinase C inhibitor was similar to that of H-7. Hydroxy H-7 inhibited the enzyme in a competitive manner with ATP, the Ki value was 23 microM, and exhibited a fluorescence property with a maximum emission wavelength of 444 nm excited at 350 nm. Fluoromicroscopical investigations revealed that hydroxy H-7 penetrated the cell membrane and was distributed mainly in the cytoplasm.

1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine↗
PubMed1988

Inhibitory effects of aminoglycosides on renal protein phosphorylation by protein kinase C.

Aminoglycosides such as neomycin are commonly prescribed antibiotics; however, there is associated serious damage to the kidney. We examined the effect of aminoglycoside antibiotics on renal protein phosphorylation and found that neomycin selectively inhibited Ca++-activated, phospholipid-dependent phosphorylation of 88-kDa protein in cell lysates of the rabbit kidney. Fifty percent inhibition of phosphorylation of this protein occurred with 5 X 10(-5) to 1 X 10(-4) M neomycin. In living PtK2 cells, neomycin dose-dependently inhibited 12-O-tetradecanoyl-phorbol-13-acetate-induced phosphorylation of 88 K Da protein. This drug also inhibited phosphorylation of exogenous protein catalyzed by protein kinase C, isolated from rabbit kidney in vitro. In contrast, neomycin had little or no inhibitory effect on cyclic GMP-dependent protein kinase, cyclic AMP-dependent protein kinase, casein kinase I, casein kinase II and Ca++-calmodulin-dependent myosin light chain kinase. Whereas activity of protein kinase C was inhibited 65% by neomycin (0.1 mM) at pH 5 to 7, inhibition decreases to 33% at pH 8 and to zero at pH 9. The potencies of a series of aminoglycoside antibiotics to inhibit the kinase agreed well with number of ionizable amino groups of compounds (gamma = 0.99) and this also approximates their known nephrotoxic potential; amikacin less than or equal to kanamycin less than gentamycin less than or equal to tobramycin less than neomycin. As aminoglycoside antibiotics present in the kidney after administration of toxicological doses (10(-2) M) will inhibit the effects of protein kinase C, the aminoglycoside antibiotics-induced nephrotoxicity is discussed in relation to inhibition of intracellular protein kinase C.

Aminoglycosides↗
PubMed1988

[Clinical studies on endometrial cancer].

One hundred and fifty-two cases with endometrial carcinoma were treated in our clinic between 1973 and 1985. The average age was 55.89 years, and the age range was from 29 to 76 years. Thirty-six cases (23.7%) were nulligravidas and 42 cases (27.6%) were nulliparas. One hundred and ten cases (72.4%) were postmenopausal and the average age at menopause was 49.1 years. The most frequent chief complaint was atypical genital bleeding which was noted in 128 cases (84.2%). The result of a cytologic examination of the uterine cervix was positive in 50.7% and suspicious in 16.9% but results for the endometrium were positive in 63% and suspicious in 21.9%. The cases in this study were classified into 88 cases (57.9%) of T1a, 36 cases (23.7%) of T1b, 12 cases (7.9%) of T2, 3 cases (2.0%) of T3 and 1 case (0.7%) of T4. As to the postoperative diagnosis, there were classified into 86 cases (56.6%) of pT1a, 30 cases (19.7%) of pT1b, 19 cases (12.5%) of pT2, 10 cases (6.6%) of pT3 and 5 cases (3.3%) of pT4. Histopathologically almost all 140 cases (91.41%) were of adenocarcinoma and classified into 88 cases (57.9%) of G1, 39 cases (25.7%) of G2 and 13 cases (8.6%) of G3. The cumulative survival rates after Kaplan-Meier were 95% in pT1a cases, 75.6% in pT1b cases, 67.3% in pT2 cases, 42.2% in pT3 cases and 0% in pT4 cases.

Adenocarcinoma↗
PubMed1988

DNA analysis of seven patients with hemophilia B who have anti-factor IX antibodies: relationship to clinical manifestations and evidence that the abnormal gene was inherited.

We have investigated genomic DNA samples of 24 patients with hemophilia B (factor IX deficiency), including seven patients with anti-factor IX antibodies (inhibitors), by molecular probes. Seventeen patients without inhibitors against factor IX and three patients with inhibitor showed no abnormalities in their restriction fragments generated by digestions of the genomic DNA by BamHl, EcoRl, Mspl, or Taql and hybridized with a factor IX cDNA probe (pHFIX). The remaining four patients with inhibitors were found to have gross deletions of the factor IX gene. Among those four patients, two were from the same family. Quantitative Southern blotting clearly showed that the abnormal gene was inherited in this family. DNA from the mother of another patient with deletion of the factor IX gene showed normal gene dosage, indicating that the mutation must have occurred at the mother's germ cells. The genomic DNA samples of four patients with gross factor IX gene deletions were found to lack the entire factor IX gene as analyzed with a factor IX cDNA as well as with a 3'-genomic factor IX fragment as probes. The hypoxanthine phosphoribosyltransferase (HPRT) gene probe, however, was found to hybridize with all of these DNA samples, indicating that the deletions in these genomic DNA samples had not extended to the region containing the HPRT gene locus in q27 proximal to the factor IX gene locus on the X chromosome. Several clinical characteristics were compared between inhibitor cases with gene deletion and inhibitor cases without obvious gene deletion.(ABSTRACT TRUNCATED AT 250 WORDS)

Chromosome Deletion↗
PubMed1987

Decreased phorbol ester receptor and protein kinase C in P388 murine leukemic cells resistant to etoposide.

A variant P388 murine leukemic cell resistant to 4'-demethylepipodophyllotoxin-9-(4,6-O-ethylidene)-beta-D-glucopyr anoside (etoposide) (VP-16-213) was cloned. The variant P388/VP-16 cell line was 159-fold resistant to 4'-demethylepipodophyllotoxin-9-(4,6-O-ethylidene)-beta-D- glucopyranoside and showed cross-resistance to vincristine (18.9-fold) and Adriamycin (522.9-fold), determined by comparing the 50% inhibitory concentrations in a 48-h growth inhibition assay. To identify the possible role of Ca2+-phospholipid-dependent protein kinase (protein kinase C) in this drug resistance, we studied the specific phorbol ester binding component and protein kinase C in the parent and drug-resistant sublines of P388 cells. The phorbol ester receptor, as expressed by the numbers of sites per cell, significantly decreased in P388/VP-16 (57.6% of control). Scatchard analysis revealed that the variant contained a single class of binding sites. However, no difference was observed in the dissociation constants (Kd), thereby suggesting much the same affinity of receptors between the two lines. Phorbol diester analogues inhibited [20-3H]phorbol-12,13-dibutyrate binding of both the variant and control cell lines, in a stereospecific manner and consistent with their binding potency. The activity of protein kinase C, which is related to the phorbol ester receptor, significantly decreased in the variant cell. The enzyme activity, particularly in the membrane fraction of P388/VP-16 cells, was remarkably decreased. These data suggest that the decrease in the specific phorbol diester receptor and protein kinase C in the variant cells might correlate with the pleiotropic drug resistance.

Animals↗