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Biomedical subjects

K Inoue

Publications and source records attributed to K Inoue.

At least 1,081 records · Page 60Linked to original sources
PubMed1996

Systemic hemodynamics and serum nitrate levels in patients undergoing endoscopic variceal ligation.

The effects of endoscopic variceal ligation (EVL) on systemic hemodynamics are unknown. This study was conducted to determine whether the obliteration of portal-systemic collaterals by EVL affects systemic hemodynamics and serum nitrate concentrations in patients with compensated cirrhosis. We measured systemic and hepatic hemodynamics, azygos vein blood flow (AzBF), and serum nitrate concentrations before and immediately following EVL. A prompt decrease in left ventricular end-diastolic volume (LVEDV), stroke volume, cardiac index (CI), and an increase in systemic vascular resistance index (SVRI) were observed following variceal ligation. The reduction in LVEDV was associated with a decline in CI with a rise in SVRI. There was also a prompt reduction in serum nitrate concentration following variceal ligation. AzBF also significantly decreased following variceal ligation. These findings indicate that EVL decreased cardiac output via a reduction in cardiac preload (central venous return). Blood flow through portal-systemic collaterals has an important role in the enhanced cardiac preload of cirrhotic patients. The sudden decrease in serum nitrate concentrations suggests that endogenous nitric oxide may be involved in the regulation of systemic hemodynamics in patients with compensated cirrhosis.

Aldosterone↗
PubMed1996

Real time observation of the binding of herpes simplex virus type 1 (HSV-1) to immobilized heparan sulfate and the neutralization of HSV-1 by sulfonated human immunoglobulin.

A real time biomolecular interaction assay system involving an optical sensor was applied to quantitative analysis of the binding of herpes simplex virus type 1 (HSV-1) to immobilized heparan sulfate, a cellular receptor component of HSV-1, and the neutralization antibody titer against this virus with a commercially available sulfonated human immunoglobulin preparation. The virus titer in a viral solution and the neutralizing antibody titer in the human immunoglobulin preparation could be successfully estimated in a short time with this system without any difficult cell culture.

Animals↗
PubMed1996

Histologic and biochemical analysis of osteogenic capacity of vascularized periosteum.

We analyzed the osteogenic capacity of the vascularized periosteum histologically and biochemically using a new experimental model of the vascularized tibial periosteum-hydroxyapatite composite in rats. Bone formation was observed not only on the surface but also in the pores of the hydroxyapatite at 4 weeks. The alkaline phosphatase activity increased to a peak at 2 weeks, with half this activity maintained until 8 weeks. The bone-specific bone gla protein content increased constantly as time passed. On the other hand, in the vascularized fascia-hydroxyapatite composite group (control group), no bone formation was observed histologically, alkaline phospatase activity was low, and bone gla protein content was very low. These results indicate that (1) the vascularized periosteum has the most significant osteogenic capaacity at 2 weeks, with a constant level of the activity maintained thereafter, (2) it forms new bone soon after operation, and (3) the amount of bone increases as time passes.

Alkaline Phosphatase↗
PubMed1996

CYP2D6 is the principal cytochrome P450 responsible for metabolism of the histamine H1 antagonist promethazine in human liver microsomes.

To determine which cytochrome P450 form is involved in the promethazine [10-(2-dimethylaminopropyl) phenothiazine] metabolism, in vitro analysis using human liver microsomes were performed. Promethazine was mainly biotransformed to ring-hydroxylated, S-oxidized and N-demethylated metabolites. The promethazine hydroxylase in human liver microsomes was inhibited by SKF-525A, propranolol, sparteine, quinidine and anti-CYP2D6 serum suggesting involvement of a P450 related to CYP2D6. Lineweaver-Burk plots for the hydroxylation, S-oxidation and N-demethylation indicated that the hydroxylation occurred with a low K(m) value in human liver microsomes. Microsomes from genetically-engineered human B-lymphoblastoid cells expressing CYP2D6 hydroxylated promethazine most efficiently as compared to other P450 forms, indicating that it was the principal P450 responsible for the metabolism of promethazine in human liver microsomes. The inhibition of CYP2D6-catalysed bufuralol 1'-hydroxylase by various histamine H3 antagonists including promethazine suggested that promethazine and some other histamine H1 antagonists could be inhibitors of this P450 in human liver microsomes.

Antibodies↗
PubMed1996

Simple method for detection of Clostridium botulinum type A to F neurotoxin genes by ploymerase chain reaction.

A polymerase chain reaction (PCR)-based method was established to detect each type of neurotoxin genes of Clostridium botulinum types A to F by employing the oligonucleotide primer sets corresponding to special regions of the light chains of the neurotoxins. In this procedure, the PCR products were easily confirmed by restriction enzyme digestion profiles, and as little as 2.5 pg of template DNAs from toxigenic strains could be detected. The specific PCR products were obtained from toxigenic C. botulinum types A to F, a type E toxin-producing C. butyricum strain, and a type F toxin-producing C. baratii strain, but no PCR product was detected in nontoxigenic strains of C. botulinum and other clostridial species. The neurotoxin genes were also detected in food products of a seasoned dry salmon and a fermented fish (Izushi) which had caused type E outbreaks of botulism. Therefore, it is concluded that this PCR-based detection method can be used for the rapid diagnosis of botulism.

Base Sequence↗
PubMed1996

Carrier identification of X-linked adrenoleukodystrophy by measurement of very long chain fatty acids and lignoceric acid oxidation.

Because identification of carriers of X-linked adrenoleukodystrophy (ALD) results in 5-15% false negatives with very long chain fatty acids (VLCFA) assay in plasma, and mutation analysis of plasma VLCFA combined with of the ALD gene is not always practical, we studied whether the analysis of plasma VLCFA combined with lignoceric acid oxidation study in fibroblasts could improve the rate of carrier detection. Lignoceric acid oxidation was abnormal in 19 out of 19 patients (ALD or adrenomyeloneuropathy) and in three out of three obligate heterozygous women. Among ten women at risk of being a carrier, three women who had normal plasma VLCFA had abnormal lignoceric acid oxidation in fibroblasts. These data suggest that this combined biochemical procedure may help to improve carrier detection in families when the ALD gene mutation has not been identified.

Adrenoleukodystrophy↗
PubMed1996

Potentiation by cadmium ion of ATP-evoked dopamine release in rat phaeochromocytoma cells.

1. The effects of cadmium ion (Cd2+) on release of dopamine and on an inward current evoked by extracellular ATP were investigated in rat phaeochromocytoma PC12 cells. 2. Cd2+ (100 microM-3 mM) potentiated the dopamine release evoked by 30 microM ATP from the cells. Cd2+ (100 microM) shifted the concentration-response curve of ATP-evoked dopamine release to the left without affecting the maximal response. 3. Suramin (30 microM) completely abolished the dopamine release evoked by 30 microM ATP but only partially inhibited the release evoked by 100 microM ATP consistent with its role as a competitive antagonist. The response evoked by 30 microM ATP in the presence of Cd2+ (300 microM) was comparable to that observed with 100 microM ATP alone; however, only the former was almost completely inhibited by suramin. 4. Cd2+ (100 microM) potentiated an inward current activated by 30 microM ATP alone. A higher concentration of Cd2+ (300 microM) had a smaller effect on amplitude potentiation but significantly prolonged the duration of the current. 5. The time-course of the ATP-evoked dopamine release was investigated using a real-time monitoring system for dopamine release. Although Cd2+ (300 microM) had little effect on the time-course of activation the ATP-evoked dopamine release, it produced a long-lasting dopamine release which slowly returned to the baseline. 6. Taken together, these observations suggest that Cd2+ enhances ATP-evoked dopamine release by affecting P2-purinoceptor/channels. The enhancement may be attributed to a Cd(2+)-dependent increase in sensitivity to ATP.

Adenosine Triphosphate↗
PubMed1996

Favorable effect of new artificial liver support on survival of patients with fulminant hepatic failure.

The two most serious symptoms of fulminant hepatic failure are bleeding and hepatic coma. To overcome these problems, we developed an artificial liver support system comprising a combination of plasma exchange and hemodiafiltration using a high performance membrane. We treated 67 patients with fulminant hepatic failure. Of these, 65 patients (97.0%) regained normal consciousness, and 55 patients (80.9%) were kept alert as long as we continued to apply this system. All 7 patients (100%) with fulminant hepatitis caused by hepatitis A virus infection and 9 of 12 patients (75%) with fulminant hepatitis caused by acute hepatitis B (HB) virus infection survived. In addition, 7 of 15 HB virus carriers (46.7%) who developed fulminant hepatitis and 11 of 29 patients (37.9%) with fulminant hepatitis caused by non-A, non-B hepatitis viruses survived. The overall survival rate was 37 of 67 patients (55.2%). Our artificial liver support system allows as high a survival rate as liver transplantation.

Aged↗
PubMed1996

[Detection of chlamydial antibodies by immunoblotting technique--in comparison of three commercially available test kits and micro-IF].

We reexamined the assay conditions of immunoblotting (W-B) technique to detect antibodies (IgG and IgA) to C. trachomatis serovar L2. Partially purified Chlamydia (35% urografin) was used as the antigen. The marker bands of W-B for positive and/or negative are major outer membrane protein (MOMP) band and either one or more bands staining in 40-62 KDa area. We then compared the sensitivity and specificity of three commercially available test kits and micro-IF by using W-B as a standard. The kits compared were sero-IPALISA-IgG-IgA, IP-Azyme-IgG-IgA and HITAZYME-IgG-IgA, and micro-IF. Serum samples were collected from the outpatient departments of gynecology, urology, and internal medicine and pediatrics. The results of agreement between W-B and test kits in IgG detection were as follows: in sero-IPALISA, total agreement was 85.7%, positive agreement 83.3%, negative agreement 90.9% and in IPAzyme: 85.7%, 83.3%, 90.9%, respectively and in HITAZYME: 82.9%, 75%, 100%, respectively. These results are almost the same as micro-IF: 85.7%, 79.2%, 100%, respectively. These kits may have relatively high sensitivity and specificity in IgG antibody detection. In IgA detection, the total agreement between W-B and sero-IPALISA was 82.9%, positive agreement 100%, negative agreement 66.7%, in IPAzyme: 77.1%, 58.8%, 94.4%, respectively and in HITAZYME: 65.7%, 64.7%, 66.7%, respectively and in micro-IF: 82.9%, 70.6%, 94.9%, respectively. Although these agreements are not so high in IgA detection, the W-B technique gives fairly consistent results as well as those kits. This indicates that W-B technique with MOMP and other protein bands (40-62 KDa) as marker for positive reaction is a useful method for detection of chlamydial antibodies.

Antibodies, Bacterial↗
PubMed1996

Molecular cloning of the gene encoding the mosaic neurotoxin, composed of parts of botulinum neurotoxin types C1 and D, and PCR detection of this gene from Clostridium botulinum type C organisms.

The DNA fragment common to the genes encoding botulinum neurotoxin types C1 (BN/C1) and D (BN/D) was amplified by PCR from the culture supernatant of Clostridium botulinum type C strain 6813 (C6813) that was treated with either DNase I or proteinase K but not from the supernatant that was treated with both DNase I and proteinase K, suggesting the neurotoxin gene is located on a certain bacteriophage DNA. Thus, to isolate the neurotoxin gene, we performed PCR with the culture supernatant of C6813 and seven primer pairs designed from the genes encoding BN/C1 and BN/D. The coding region in the connected sequence encodes a neurotoxin composed of 1,280 amino acids with a molecular weight of 147,817. The neurotoxin from C6813 has 95% amino acid identity to BN/C1, except for its C-terminal one-third, which is quite similar to the C-terminal one-third of BN/D (95% identity). When we performed PCRs with four primer pairs designed from the 5'-terminal two-thirds of the BN/C1 gene and two primers from the 3'-terminal one-third of the BN/D gene, DNA fragments of the expected sizes (0.5 to 1.3 kbp) could be amplified from C. botulinum type C strains 6812 and 6814. These results suggest that some strains of C. botulinum type C contain the gene encoding the mosaic neurotoxin composed of parts of BN/C1 and BN/D.

Amino Acid Sequence↗
PubMed1996

Molecular composition of Clostridium botulinum type A progenitor toxins.

The molecular composition of progenitor toxins produced by a Clostridium botulinum type A strain (A-NIH) was analyzed. The strain produced three types of progenitor toxins (19 S, 16 S, and 12 S) as reported previously. Purified 19 S and 16 S toxins demonstrated the same banding profiles on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), indicating that they consist of the same protein components. The nontoxic components of the 19 S and 16 S toxins are a nontoxic non-hemagglutinin (HA) (molecular mass, 120 kDa) and HA. HA could be fractionated into five subcomponents with molecular masses of 52, 35, 20, 19, and 15 kDa in the presence of 2-mercaptoethanol. The molar ratios of neurotoxins, nontoxic non-HAs, and each HA subcomponent of the 19 S and 16 S toxins showed that only HA-35 of the 19 S toxin was approximately twice the size of that of the 16 S toxin, suggesting that the 19 S toxin is a dimer of the 16 S toxin cross-linked by the 35-kDa subcomponent. The nontoxic non-HA of the 12 S toxin, but not those of the 19 S and 16 S toxins, demonstrated two bands with molecular masses of 106 and 13 kDa on SDS-PAGE with or without 2-mercaptoethanol. It was concluded from the N-terminal amino acid sequences that 106- and 13-kDa proteins were generated by a cleavage of whole nontoxic non-HA. This may explain why the 12 S and 16 S (and 19 S) toxins exist in the same culture. We also found that the HA and its 35-kDa subcomponent exist in a free state in the culture fluid along with three types of progenitor toxins.

Amino Acid Sequence↗
PubMed1996

Evaluation of L-pyrrolidonyl peptidase paper strip test for differentiation of members of the family Enterobacteriaceae, particularly Salmonella spp.

The L-pyrrolidonyl peptidase activities of 1,033 strains of the family Enterobacteriaceae were investigated by the paper strip method to evaluate their usefulness for screening those organisms, especially Salmonella cultures. We also evaluated the usefulness of indole and tryptophan deaminase paper strip tests as supplements to the L-pyrrolidonyl peptidase test for the rapid identification of Salmonella cultures. The paper strip tests are simple, and the results are obtainable within 10 min.

Amino Acid Oxidoreductases↗
PubMed1996

Vertebral growth disturbance in rapidly growing rats during 14 days of spaceflight.

The effect of 14 days of spaceflight on the vertebrae of rapidly growing rats was studied. The hardness of the vertebrae was measured with a Knoop microhardness tester, and bone mineral density was measured from X-ray photographs. Histomorphometric examination was performed with a microcomputer-aided system. No significant difference (P > 0.05) was observed between flight rats and ground controls with regard to mechanical hardness or bone mineral density. However, histological examination revealed irregular thickening of the endosteal surface of cortical bone in the flight rats, whereas it was uniform in the ground controls. The relative area of lamellar bone showed a significant reduction (P < 0.001) in the flight rats. These findings suggest that the structural disturbances were due to retardation of endosteal modeling and remodeling. We conclude that delay of vertebral maturation can occur in rapidly growing rats after even short-term exposure to microgravity.

Animals↗
PubMed1996

An adjustable-current swimming pool for the evaluation of endurance capacity of mice.

A new forced-swimming apparatus for determining maximum swimming time in mice was devised for use in the evaluation of the endurance capacity of Std and ddY and CDF1 mice after various diet and drug treatments. With the apparatus, a water current is generated by circulating water with a pump in a swimming pool. A spout and suction slit were contrived to generate a constant current while the strength of the current is regulated by a valve. The decrease in the leg-kicking intervals of mice accompanying the increase in the current speed confirmed that the workload is adjustable by regulation of the current speed. Compared with the number of forelimb strokes, that of the hindlimb kicks was greater. The swimming time until fatigue was observed to decrease with increasing current speed in the two strains of mice. As biochemical indexes, the blood lactate and muscle glycogen levels corroborated the correlation between current speed and increase in workload. These results indicate that the apparatus employed in the present study is suitable for the evaluation of the endurance capacity of mice and that is useful for detecting the effects of dietary differences and drug pretreatments on this capacity.

Animals↗
PubMed1996

Spontaneous perforated pyometra presenting as pneumoperitoneum.

BACKGROUND: Spontaneous perforated pyometra presenting as pneumoperitoneum is extremely rare. CASE REPORT: An 80-year-old Japanese female with spontaneous perforating pyometra presenting as pneumoperitoneum is reported. The patient came to our institute with severe abdominal pain. Routine abdominal examination showed muscular defense, and plain chest roentgenograms revealed infradiaphragmatic free gas. Subsequent computed tomography also demonstrated pneumoperitoneum. Laparotomy was performed on the basis of a tentative diagnosis of perforation of the gastrointestinal tract but revealed a perforated pyometra. A simple hysterectomy was performed. The histological diagnosis of the surgical specimen was acute endometritis without neoplasm. The present report is the third case of spontaneous perforated pyometra with pneumoperitoneum to date. CONCLUSION: Although uterine disease presenting as pneumoperitoneum is rare in elderly patients with an acute abdomen, the possibility of a perforated pyometra should be considered in the differential diagnosis.

Aged↗
PubMed1996

Characterization of monoclonal antibodies against (1R,2R)-cyclohexanediamine platinum(II)-DNA adduct.

Two monoclonal antibodies raised against Pt(II)(1R,2R-cyclohexanediamine)-DNA were prepared, and the specificity of the monoclonal antibodies against Pt(II)(cyclohexanediamine)-DNA derivatives was determined by competitive enzyme-linked immunosorbent assay. The binding affinity of the monoclonal antibodies is apparently influenced by the cyclohexanediamine moiety of Pt(II)(cyclohexanediamine)-DNA adducts, but the monoclonal antibodies can not bind to the low molecular analogue, Pt(II)(1R,2R-cyclohexanediamine)-d(GpG), which is the intrastrand binding model compound of Pt(II)(1R,2R-cyclohexanediamine)-DNA. Therefore, the monoclonal antibodies recognize the macromolecular parts, including DNA duplex, in addition to the cyclohexane moieties of the platinum complexes on Pt(II)(cyclohexanediamine)-DNA adducts.

Antibodies, Monoclonal↗
PubMed1996

Lysophosphatidylserine enhances exogenous type II phospholipase A2-induced activation of rat serosal mast cells.

We have previously shown that exogenous type II phospholipase A2 (PLA2) alone elicits degranulation of mast cells, including rat serosal mast cells (SMC) and mouse bone marrow-derived mast cells (BMMC). Here we report that lysophosphatidylserine (lysoPS), a co-factor for activation of rodent SMC in response to some tyrosine kinase-coupled agonists, enhanced type II PLA2-elicited histamine release from rat SMC. In contrast, mouse BMMC was insensitive to lysoPS. Our findings demonstrate a novel route for activation of SMC in that type II PLA2 can act as a direct activator of SMC with enhancement by lysoPS, which is generated from membrane phosphatidylserine possibly by the action of the same enzyme.

Animals↗