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Biomedical subjects

S Otsuji

Publications and source records attributed to S Otsuji.

At least 37 records · Page 2Linked to original sources
PubMed1992

[A case of hepatoma patient who showed an abnormally high level of serum IRI with the beads method].

A fifty four years old hepatoma patient admitted to the hospital for a surgical operation. Preoperative laboratory examination demonstrated that his serum IRI level was very high (423 microU/ml) when measured with a beads method, however RIA or a microplate method demonstrated normal values. We studied the mechanism of the discrepancy of IRI values. 1) Both the beads and microplate methods demonstrated the same IRI values when the patient's serum insulin was roughly purified with Sep-Pak. The beads method showed high IRI values in serum which passed through Sep-Pak, therefore contained no insulin. 2) The similar results were observed when the patient's serum fractionated by a gel-chromatography (Biogel P-30). The beads method demonstrated high IRI values in both insulin fractions and the fractions containing serum proteins bigger than 40,000 molecular weight. The microplate method demonstrated only one large peak of insulin. 3) When non-specific IgG of guinea pig was used as a fixed antibody instead of human insulin antibody of guinea pig that was used in the beads method, the patient's serum showed the similar values as that obtained with the beads method. We thereby concluded that the abnormal level of IRI by the beads method was derived from the unknown substance reacting with IgG of guinea pig in the patient's serum. After the surgical resection of hepatoma, the levels of IRI measured by the beads method decreased significantly, suggesting that the substance is related to hepatoma cells.

Animals↗
PubMed1992

[A case of insulin receptor abnormality (type A)].

A sixteen year old woman came to the hospital for glucosuria and amenorrhea. Physical examination demonstrated that she had hirsutism, deepening of voice, and pigmented skin in her axillary lesion which was histologically diagnosed as acanthosis nigricans. Ultrasonography showed polycystic ovaries. A diabetic pattern of 75 g oral glucose tolerance test, very high levels of serum insulin (fasting: 320, peak: 1,220 microU/ml), and hyperandrogenism characterized by increases of urine 17-KS, serum testosterone and DHEA-S were found. Both serum insulin and insulin-receptor antibodies were found to be negative. Insulin binding to both erythrocytes and cultured skin fibroblasts were significantly decreased (about 30% of normal controls). Scatchard plot analysis demonstrated decreased number of insulin receptors to about 30% of the normal controls. We therefore diagnosed that she had insulin receptor abnormality, Type A in Kahn's classification.

Acanthosis Nigricans↗
PubMed1991

[Nosocomial infections due to methicillin-resistant S. aureus (MRSA) at the Kagoshima University Hospital (1). Coagulase typing of MRSA].

Nosocomial infection due to MRSA at the Kagoshima University Hospital and their coagulase typing were examined using S. aureus (349 strains) clinically isolated in 1989. The results were as follows: 1) S. aureus consisted of 43.6% of MRSA and 56.7% of Methicillin-sensitive S. aureus (MSSA). 2) MRSAs were recovered most frequently from the specimens from the respiratory tract (47.8%). 3) The isolation of MRSA gradually increased in frequency from January to August; however, that of MSSA did not show a similar tendency. 4) The isolation of MRSA was higher in frequency in the surgical wards, the ICU and the pediatric ward. 5) When classified into 8 coagulase types, MRSAs (133 strains) consisted of only 3 types (54.1% of type II, 32.3% of type VII, and 12.0% of type IV), whereas MSSAs contained all coagulase types. As compared with the results of coagulase typing at other institutions, the incidence of type VII was much more frequent at our hospital. 6) A few coagulase types of MRSA appeared in each ward. Also, type II MRSA strains increased in June and July, and type VII MRSA increased in August. However, MSSA strains did not show any similar tendency.

Bacterial Typing Techniques↗
PubMed1991

[Nosocomial infections due to methicillin-resistant S. aureus (MRSA) at the Kagoshima University Hospital (2). Susceptibility to antibiotics].

Susceptibility to antibiotics of 123 MRSA strains isolated at the Kagoshima University Hospital in 1989 was examined. The results were as follows: 1) Susceptibility of MRSA strains was excellent to VCM, MINO, and RFP, followed by IMP, CLDM, and CPFX. However, most strains were resistant to PCG, MPIPC, ABPC, CET, CMZ, CZON, GM, and AMK. 2) Five strains highly resistant to RFP were isolated. Three of these strains were isolated on the ward for tuberculosis. This suggests easy development of resistance to RFP by MRSA. 3) Differences in susceptibility to antibiotics between coagulase type II and type VII strains were examined. The cumulative percentage of type II strains susceptible to CLDM (MICs less than 0.5 microgram/ml) was 15.3%, and that of type VII was 45.2%. Strains resistant to CLDM were more frequently isolated among type II than among type VII strains (p less than 0.001). A similar relationship between strains and antibiotics was also found with EM and CPEX. On the other hand, the cumulative percentage of type II strains susceptible to AMK (MICs less than 25 micrograms/ml) was 89.7%, and that of type VII was 9.7%. Strains resistant to AMK were more frequently isolated among type VII than among type II strains (p less than 0.001).

Anti-Bacterial Agents↗
PubMed1991

[Introductory remarks for symposium on "biological membranes of blood cells"].

Recent progress on studies of structure and functions of biological membranes has been successfully achieved. Various topics on membrane abnormalities in blood cells were selected for this symposium. First of all, in red cell disorders, abnormal red cell morphology is crucial for identification of various kinds of hemolytic anemias. In addition, cytoskeletal protein abnormalities in red cell membrane were reviewed, especially on the pathogenesis of hereditary elliptocytosis and hereditary spherocytosis. Abnormal red cell membrane fluidity was also discussed in relation to insulin receptor on red cell membranes in diabetic patients. A role of growth factors, especially G-CSF and GM-CSF, on granulocytes was described from the standpoint of membrane receptors. Platelets demonstrate various glycoproteins (GP) on their cell membrane surfaces. Abnormalities on these glycoproteins, such as GP IIb-IIIa and GP Ib-IX complex, were observed in thromboasthenia and Bernard-Soullier syndrome. In addition, arachidonic acid cascade is important in platelet functions, especially for signal transduction of extracellular messages through plasma membranes. Finally, a new application of a liposome immune lysis assay was introduced for laboratory determinations of complements, antibodies, and various marker proteins. The symposium will be extremely helpful for further understanding of the structure-function relationship and of the various disorders of blood cell membranes.

Blood Cells↗
PubMed1990

A rapid turbidimetric immunoassay for serum antistreptolysin-O.

We designed a rapid, quantitative turbidimetric immunoassay for serum antistreptolysin-O (ASO), based on the immunoprecipitating velocity with purified streptolysin-O (SLO), which can be measured turbidimetrically at 340 nm. The assay system involves polyoxyethylene mono-p-nonylphenyl ether to accelerate and enhance the immunoprecipitation reaction. No antibody excess phenomenon is observed, up to 1,700 U/ml. Grossly icteric sera can be assayed. Patient sera gave acceptable results. Free cholesterol offers no interference. The standard curve is linear for values up to 800 U/ml. Results measured by the proposed method and those by the conventional microtitration method correlated well. Analytical recovery averaged 90.1% (80.3%-102.5%). Within-run and day-to-day CVs ranged from 2.1% to 3.5% and from 2.5% to 6.9%, respectively. Working SLO is stable for at least 15 days at 4 degrees C. The proposed method has proven to be superior to other available methods for measuring serum ASO.

Antistreptolysin↗
PubMed1988

A new enzymatic approach for estimating total and direct bilirubin.

We designed an enzymatic assay for total (TBil) and direct bilirubin (DBil), the principle of which involves measuring the decrease in absorbance at 450 nm produced by bilirubin oxidase from Myrothecium verrucaria. Since TBil and DBil are oxidized at pH 7.2 and 3.7, respectively, the degree of bilirubin oxidation is measurable in each case. An analysis of bilirubin by high-performance liquid chromatography, before and after the enzymatic reaction with bilirubin oxidase, verified the specificity of the enzyme. The results obtained using this method varied linearly with TBil and DBil concentrations up to at least 250 mg/L and 150 mg/L, respectively. Reducing substances, commonly used anticoagulants and hemoglobin showed no apparent interference. The degree of day-to-day precision (CV) for TBil and DBil ranged from 1.2% (206.2 mg/L) to 10.6% (3.5 mg/L) and from 1.8% (84.3 mg/L) to 12.4% (2.1 mg/L), respectively. Values measured using this new method correlated well with those obtained by Malloy-Evelyn's method and the slide method employing the Kodak Ektachem analyzer.

Bilirubin↗
PubMed1988

Albumin prevents erythrocyte crenation in patients undergoing extracorporeal circulation.

Free fatty acid-induced massive erythrocyte crenation has been reported in patients undergoing extracorporeal circulation (EC), suggesting that the crenated cells impair microcirculatory flow and tissue oxygenation during and after the operation. Effect of albumin administration on erythrocyte crenation was examined in 18 patients undergoing coronary bypass operation: 9 patients were given 25 g and another 9 patients were given 50 g of human albumin as a part of the priming solution. Erythrocyte crenation was almost completely prevented in the patients given 50 g albumin. Crenated erythrocytes during EC were 3.6 +/- 2.3% (mean +/- SD) of all erythrocytes and 2.8 +/- 3.7% after EC. This was significantly lower than in patients without albumin administration (63.4 +/- 34.0% during EC and 28.6 +/- 33.3% after EC, n = 20). But the effect was less striking in the patients given 25 g albumin, 32.4 +/- 39.1% during and 28.3 +/- 40.8% after EC. Maintaining an adequate level of plasma albumin is important in preventing erythrocyte crenation during EC, improving microcirculatory flow in patients undergoing open heart surgery.

Aged↗
PubMed1987

Erythrocyte crenation induced by free fatty acids in patients undergoing extracorporeal circulation.

Normal erythrocyte morphology is necessary for proper distribution of blood-flow in the microcirculation. Erythrocyte shape was studied in 20 patients undergoing extracorporeal circulation (EC) during coronary bypass surgery. Crenated erythrocytes comprised a mean 64% of all erythrocytes during and 29% after EC. Free fatty acid (FFA) content of the erythrocyte membranes was significantly increased at both times, and FFA content correlated with the proportion of crenated erythrocytes. Washing crenated cells with defatted albumin solution transformed them back to discocytes, simultaneously removing more FFA than that removed from pre-EC discocytes. The plasma FFA to albumin ratio became disproportionately increased during EC; the increased level correlated with severity of erythrocyte crenation. Maintaining a higher level of albumin during EC by adding 50 g human albumin to the extracorporeal system prevented erythrocyte crenation. Entry of plasma FFA not bound to albumin into erythrocyte membranes during EC causes massive erythrocyte crenation.

Adult↗
PubMed1987

A simple enzymatic differential assay for diamines, spermidine, and spermine in urine and blood.

This rapid, reliable enzymatic differential assay method for diamines (putrescine plus cadaverine), spermidine, and spermine in urine and blood is suitable for practical routine use and does not require special and expensive equipment. The method is based on a combination of the substrate specificities of two amine oxidases, i.e., polyamine oxidase from A. terreus and putrescine oxidase from M. rubens. Quinone dye, derived from hydrogen peroxide generated in each of three end-point reactions, is measured spectrophotometrically at 555 nm, and the amounts of the respective amines are simply calculated. Analytical recoveries and precisions are acceptable. The proposed method produces results which correlate with high-performance liquid chromatography. Twenty or more assays can be done within 3 hr by one technician.

Chromatography, High Pressure Liquid↗
PubMed1987

A rapid enzymatic assay for total blood polyamines.

We have designed a rapid, reliable enzymatic assay for total blood polyamines, as based on the combination of substrate specificity of polyamine oxidase (PAO) from Aspergillus terreus and putrescine oxidase (PUO) from Micrococcus rubens. Quinone dye, derived from hydrogen peroxide generated in the oxidation reaction, is measured spectrophotometrically at 555 nm, and total amounts of putrescine (Put), cadaverine (Cad), spermidine (Spd), and spermine (Spm) can be readily determined. The minimum detection limit is about 0.4 mumol/L whole blood. Bilirubin, hemoglobin, reducing substances, and anticoagulants show no apparent interference. Analytical recovery averaged 98.5%. Within-run and between-day precisions ranged from 1.02% (61.04 mumol/L) to 2.84% (13.07) and 1.54% (50.67) to 3.27% (14.74), respectively. Results obtained with this method and those by high-performance liquid chromatography (HPLC) (r = 0.955) or the enzymatic differential method (r = 0.944) correlated well. In our opinion, this method is superior to other assays being used to determine blood polyamines.

Adult↗
PubMed1986

Dietary sardine oil increases erythrocyte membrane fluidity in diabetic patients.

The effects of dietary sardine oil rich in eicosapentaenoic acid, C20:5 (EPA), on erythrocyte membrane fluidity and membrane and plasma lipids were investigated in diabetic and control subjects. Before consumption of this oil, the levels of erythrocyte membrane fluidity were lower in the diabetic subjects, as noted in our previous work (Diabetes 1983; 32:585-91). Decreased membrane polyunsaturated fatty acid contents were evident. Daily consumption of 2700 mg of sardine oil for 8 wk increased erythrocyte membrane fluidity, as determined by electron spin resonance using the 12- or 16-stearic acid label. This increase was seen after 4 wk, and the level remained elevated for 8 wk. Membrane EPA of phospholipid acyl-chains significantly increased after 4 wk and was even more apparent after 8 wk. Membrane-free cholesterol to phospholipid molar ratios significantly decreased after 8 wk. Both the diabetic and normal subjects responded to the sardine oil in the same way. After feeding with sardine oil, there no longer were differences in erythrocyte membrane fluidity between the normal and diabetic subjects. We propose that improvement in membrane fluidity may contribute to the amelioration of altered cell membrane functions in diabetic patients.

Adult↗
PubMed1985

An enzymatic differential assay for urinary diamines, spermidine, and spermine.

Subsequent to the hydrolysis of urinary conjugated amines by heating with hydrochloric acid, free amines were isolated by cation-exchange chromatography. SPD and SPM in an aliquot of amine extract were first oxidized by PAO from Penicillium chrysogenum, producing PUT and hydrogen peroxide. DIAs, which consist of the initially present DIAs plus PUT produced by PAO, were subsequently oxidized by PUO from Micrococcus rubens, producing hydrogen peroxide. In an another aliquot of the amine extract DIAs and SPD were oxidized by PUO, producing hydrogen peroxide. Quinone dye, derived from hydrogen peroxide generated in each end-point reaction, was measured spectrophotometrically at 555 nm, and the amounts of the respective amines in urine were calculated. Significantly elevated levels of DIA, SPD, SPM, and an elevated DIA to SPD ratio were found in urine from 46 cancer patients, as compared to 34 normal control subjects. An increase in DIA and the ratio of DIA to SPD was found at clinical tumor stage I of the alimentary tract. The levels of DIA remained fairly constant and the ratio of DIA to SPD was consistently decreased with advancing clinical tumor stages. In patients who had undergone curative resection, there were greater decreasing rates (80% of cases for DIA and 80% for SPD) than in patients who had undergone noncurative resection (45.5% for DIA and 36.4% for SPD).

Diamines↗
PubMed1985

Qualitative and quantitative analysis of erythrocyte surface membrane sialyl residues using affinity cytochemistry with special reference to diabetic patients.

Erythrocyte surface membrane sialyl residues were investigated by means of affinity cytochemistry using the avidin-biotin complex technique. Mild oxidation with the periodate (MO)-biotin hydrazide (BHZ)-ferritin avidin conjugate (FAv) sequence revealed numerous ferritin particles on erythrocytes from healthy donors. The ferritin particles attached on the perpendicularly sectioned membrane were seen at an average distance of 10 to 12 nm from the outer dense leaflet of the cell membrane. Pretreatment with neuraminidase followed by the MO-BHZ-FAv sequence almost eliminated erythrocyte ferritin labeling. Erythrocytes from diabetic patients showed less dense ferritin labeling compared with those from healthy donors. Quantiative analysis of sialyl residues demonstrated a marked reduction in ferritin labeling of erythrocytes from diabetic patients which was significantly less (p less than 0.01) than that of erythrocytes from healthy donors. This observation supports previous biochemical data demonstrating lower levels of surface membrane negative charge and sialyl residues on erythrocytes from patients with diabetes mellitus.

Adult↗
PubMed1985

Erythrocyte membrane fluidity decreased in uremic hemodialyzed patients.

Erythrocyte membrane fluidity was studied by means of electron spin resonance in 15 uremic, hemodialyzed patients and 14 normal subjects. Erythrocyte membrane fluidity determined using a 16-nitroxide stearic acid spin label probe was of a significantly lower level in the uremic patients, when compared with normal control subjects. Alterations in molar ratios of membrane free cholesterol to phospholipid are probably not a principal factor contributing to this change in fluidity. Significant decreases of phosphatidylcholine and molar ratios of phosphatidylcholine to sphingomyelin were noted in the erythrocyte membrane of uremic patients, and these alterations may relate to the fluidity change.

Adult↗