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Biomedical subjects

H Kodama

Publications and source records attributed to H Kodama.

At least 415 records · Page 23Linked to original sources

Detection of salmonid herpesvirus (Oncorhynchus masou virus) in fish by Southern-blot technique.

To detect Oncorhynchus masou virus (OMV) in salmonid fish, we prepared recombinant plasmids from two distinct BamHI digests of OMV DNA as a probe. Viral DNAs could be detected two weeks earlier before the virus was isolated from asymptomatic or dead fish. Using the probe, viral DNAs were detected from fish reared in fish hatcheries or fish captured in a river in Hokkaido. The sensitivity limit of detection was estimated to be 10 copies of viral DNA per cell.

Animals↗

Effect of intestinal microflora on digestible energy and fiber digestion in chickens fed a high-fiber diet.

The effect of intestinal microflora on digestible energy (DE) value and fiber digestion was studied in single-comb White Leghorn chickens fed a low-fiber diet (experiment 1) or a high-fiber diet with low or adequate metabolizable energy (ME) value (experiment 2). Fecal energy excretion was calculated from the difference between total energy excretion in urinary and fecal droppings and urinary energy excretion, which was estimated from the energy values for individual urinary nitrogenous compounds extracted with Li2CO3. When the birds were fed the low-fiber diet, no differences in growth, DE, or ME were observed between germ-free and conventional environments. Of birds fed the high-fiber diet, growth of those in the conventional environment was similar to that of the birds in the germ-free environment at the adequate ME value, whereas birds in the conventional environment grew faster than the birds in the germ-free environment at the low ME value. Changes in observed dietary ME values of the high-fiber diets, being higher in birds in the conventional environment than in birds in the germ-free environment (experiment 2), were almost entirely accounted for by those in dietary DE values, most of which was contributed by crude fiber digestion. It was concluded, therefore, that by means of fiber digestion, the intestinal microflora may benefit the host bird by supplying extra energy, which would result in growth promotion, particularly when the bird is deficient in energy.

Animals↗

Comparison of different Oncorhynchus masou virus (OMV) strains by DNA restriction endonuclease cleavage analysis.

Seven strains of Oncorhynchus masou virus (OMV) genomes were analyzed with the restriction endonucleases BamHI, EcoRI, HindIII and SmaI. The restriction patterns of OMV strain DNAs were divided into four groups. Restriction profiles of high passage strains (00-7812, 65th passage, and H-83, 60th passage) were different from those of low passage strains (00-7812, 8th passage, and H-83, 6th passage) when digested with BamHI, HindIII and SmaI. However, no difference was observed between the restriction patterns of high and low passage viral DNA with EcoRI. There was no distinct difference observed between the restriction patterns of tumor tissue-derived and coelomic fluid-derived strains. By using 32P-labelled DNA of standard OMV (strain 00-7812) as a probe, most of the fragments of other OMV strain DNAs were hybridized.

Animals↗

An improved ELISA system for the measurement of IgG antibodies against pertussis toxin (PT) and filamentous hemagglutinin (FHA) in human sera.

We have developed a modified assay method for the measurement of antibodies against PT and FHA. Polystyrene balls coated with purified PT and FHA were incubated with standard or test sera, washed, incubated with goat anti-human IgG F(ab')2 conjugated with horseradish peroxidase and then washed again. Peroxidase activity was then measured by color development using o-phenylenediamine. The anti-PT and anti-FHA IgG titers were determined using calibration curves. The assay standard curves for anti-PT and anti-FHA were both linear from 1 to 100 ELISA units per milliliter (EU ml-1), and the assay sensitivity was calculated to be 1 EU ml-1 in both cases. Intra- and inter-assay CVs were both less than 10%. Inter-laboratory assay CVs calculated from the results from four different facilities were 3.3-34.6% (average 14.5%) and 2.1-30.3% (average 13.6%) for anti-PT and anti-FHA IgG, respectively, and the correlation coefficient between the Chinese hamster ovary (CHO) cell assay and the anti-PT PS-ball ELISA assay was 0.959. Using this assay method we obtained in a highly sensitive and reproducible measurement of IgG antibodies against PT and FHA in human sera. This assay should be especially useful for the determination of low concentrations of these antibodies.

Adhesins, Bacterial↗

Activation of rainbow trout complement by C-reactive protein.

The activation of the complement system of rainbow trout by trout C-reactive protein (CRP) was investigated. Complement fixation tests were performed by using rabbit hemolysin-sensitized sheep erythrocytes and rainbow trout complement. Purified CRP increased the consumption of complement in the presence of Streptococcus pneumoniae C-polysaccharide (CPS), indicating the activation of the complement system. In contrast to this, acute phase serum activated the complement in the absence of CPS. Consumption of the complement by acute-phase serum was depressed when CRP was removed from acute-phase serum by CPS-sepharose 4B affinity chromatography. The acute-phase serum, as well as CRP plus CPS, suppressed in vitro growth of Vibrio anguillarum in the presence of complement, and enhanced the phagocytosis of the bacteria by glass-adherent peritoneal exudate cells. These results indicated that CRP has a role in host defense during acute-phase response through the activation of the complement system, enhancement of phagocytosis, and suppression of bacterial growth.

Acute-Phase Reaction↗

[Changes of hypothalamo-pituitary-thyroid function after open heart surgery--especially evaluated by TRH test].

Changes of pituitary and thyroid functions after operation were studied in the cases with cardiopulmonary bypass (ECC group) and without bypass (Control group). To know mechanisms of these changes, reaction of hypothalamo-pituitary-thyroid axis by using thyrotropin-releasing hormone (TRH) test and influence of serum cortisol level were examined. Serum concentrations of free T3, free T4 and TSH decreased after open heart surgery. The levels of TSH and free T4 were recovered at the second or third post operative day in Control group, but remained low level at the third POD in ECC group. The level of free T3 continued low level at the 7th POD. The feedback mechanism of the hypothalamo-pituitary-thyroid system was not seen in this series. The response of TSH and free T3 due to TRH test decreased in ECC group, and it was suppressed more clearly in the cases of longer bypass than in those of shorter ones. Serum cortisol level was significantly increased at the 3-hours after the operation in ECC group, and was more increased in the cases of longer bypass than in those of shorter ones. The results indicate that the pituitary and thyroid function on the post bypass period remains euthyroid sick syndrome and hypothalamo-pituitary axis remains suppressed state. Those phenomenon suggest that TRH test may be useful in evaluation of the intensity of surgical stress, especially in those who undergo the cardiopulmonary bypass.

Adolescent↗

[Effects of dopamine and dobutamine on systemic hemodynamics and myocardial metabolism in children after open heart surgery].

The purpose of this study is to know the effects of Dopamine (DOA) and Dobutamine (DOB) on the systemic hemodynamics and myocardial metabolism in the acute phase after open heart surgery in children. Thirty patients with congenital heart disease were divided into following two groups. The first 14 cases were administered 5 and 10 micrograms/kg/min (gamma) of DOA, and the systemic hemodynamic and metabolic data were taken before and after the administration of the drug. The second 12 cases were given the same doses of DOB instead of DOA. DOA: The blood pressure was elevated by 10 gamma of DOA, and cardiac index (CI) and stroke volume index (SVI) rose up at both doses of DOA. On the other hand, systemic vascular resistance (SVR) and left atrial pressure (LAP) were decreased at both dosage levels. DOB: At the same dose of DOA, DOB increased HR and SVI, so CI rose up markedly. The systolic and mean blood pressure also rose up at both doses. CVP and LAP were depressed at either dosage level. SVR did not show an appreciable change. Myocardial metabolism: The two drugs tested did not exhibit the progress of anaerobic myocardial metabolism. The myocardial oxygen uptake rate increased with DOA, but decreased with DOB. This phenomena probably suggests that DOB dilates coronary vascular bed. From the above data, the following effects are expected by the use of each drug after open heart surgery in children: 1) an increase of cardiac output due to inotropic action by DOA, 2) powerful inotropic and chronotropic action by DOB.

Child↗

Synthesis and NMR study of ribo-oligonucleotides forming a hammerhead-type RNA enzyme system.

We designed a hammerhead-type ribozyme system which consists of three RNA fragments and synthesized the component and related ribo-oligonucleotides by the solid-phase phosphoramidite method using o-nitrobenzyl groups for 2'-hydroxyl protection. Improved conditions for photolytic removal of the o-nitrobenzyl groups are described. Imino proton NMR spectra of the ribozyme complex were measured. The resonances for the hydrogen-bonded imino protons were assigned by comparison with the spectra of model complexes which contain the base sequences of one or two of the stem regions. The results suggest that the complex indeed forms a hammerhead structure and the loop regions take an ordered conformation in the absence of magnesium ions.

Base Composition↗

Liquid chromatography-mass spectrometry for the qualitative analyses of iminodipeptides in the urine of patients with prolidase deficiency.

Analyses of standard iminodipeptides and iminodipeptides in the urine of patients with prolidase deficiency have been demonstrated using liquid chromatography-mass spectrometry with an atmospheric pressure ionization interface system. The separation was carried out on a reversed-phase column using 0.1% aqueous trifluoroacetic acid-methanol (70:30 or 80:20). Very intense quasi-molecular ions [( M + H]+) of various standard iminodipeptides were observed by this method. The quasi-molecular ions [M + H]+ of various iminodipeptides were also observed in the urine samples of patients with prolidase deficiency, and Gly-Pro, Ala-Pro, Val-Pro, Leu-Pro, Ile-Pro, Ser-Pro, Thr-Pro, Glu-Pro, Asp-Pro, His-Pro, Lys-Pro, Pro-Pro and Tyr-Pro as iminodipeptides containing proline with C-terminal residue and Glu-Hyp, Pro-Hyp, Ile-Hyp and Gly-Hyp as iminodipeptides containing hydroxyproline with C-terminal residue were identified in the urine of patients with prolidase deficiency.

Chromatography, High Pressure Liquid↗

Stepwise progression of B lineage differentiation supported by interleukin 7 and other stromal cell molecules.

Growth of early B precursor cells was investigated in vitro by using rIL-7 and IL-7-defective stromal cell line PA6 as separate growth signals. B cell development proceeds through three sequential stages different from the growth signal requirement. The cells in the first stage require PA6 alone for the proliferation, and differentiate into the second stage, which requires both PA6 and IL-7 for its growth. When IL-7 is available for the cells in the second stage, they proliferate extensively on the PA6 layer, and some acquire the ability to proliferate in response to IL-7 alone. This sequential change of growth signal requirement, however, does not proceed autonomously along the time schedule. The possibility that it is primarily directed by the result of Ig gene rearrangement is considered. This mode of growth control may explain why only functional B cells are selected in the error-prone process of Ig gene rearrangement during B lineage differentiation.

Animals↗

Specific inhibitory conformation of dipeptides for chymotrypsin.

Based on the analyzed conformation of a chymotrypsin inhibitor H-delta Phe-Phe-OMe, we have designed a series of diastereomeric phenylalanylphenylalanine methyl esters and derivatives as possible inhibitors. Among the peptides synthesized, H-D-Phe-L-Phe-OMe was found to be very resistant to chymotrypsin in spite of its L-Phe-OMe structure at the C-terminus. It inhibited the enzyme fairly strongly and competitively with Ki = 9.0 x 10(-5) M in the assay using Ac-Tyr-OEt as a substrate. The measurements of the NOEs in high-resolution 1H-NMR analyses indicated the presence of the hydrophobic core built by the intramolecular interaction between the D-Phe-phenyl and ester-methyl groups. It was suggested that this core interacts with the chymotrypsin S2 site (Trp215) and Phe2 with the S1 site. The backbone structure of this dipeptide was assumed to be in an inhibitory conformation that fits the active center of the enzyme.

Amino Acid Sequence↗

Characterization of prolidase I and II from erythrocytes of a control, a patient with prolidase deficiency and her mother.

Prolidase I (EC 3.4.13.9) was purified to homogeneity from the erythrocytes of a normal human (control) and the patient's mother, and prolidase II from erythrocytes of a control and the patient's mother, and prolidase from the patient's erythrocytes was also highly purified. The various properties of the patient's prolidase were compared to those of prolidase from a control and the patient's mother. Prolidase I from a control and the patient's mother had a molecular weight of about 112,000, and was composed of two subunits with an identical molecular weight of 56,000. The Km values for Gly-Pro of the control's and the patient's mother's prolidase I were 2.90 +/- 0.22 and 2.88 +/- 0.27 mM, but the Vmax values for Gly-Pro of the mother's enzyme was reduced about 30% compared to that of control enzymes (mother: 6.02 units/mg protein, control: 22.21 units/mg protein). Isoionic points of these enzymes by chromatofocusing were pH 4.6 approximately 4.7. Prolidase II from the control and the patient's mother, and the patient's prolidase had a molecular weight of about 185,000, and was composed of two subunits with an identical molecular weight of 95,000. The Km and Vmax values for various substrates of prolidase II from a control and the patient's mother, and the patient's prolidase were almost the same.

Chromatography, Gel↗

The therapeutic effect of OK-432-combined adoptive immunotherapy against liver metastases from breast cancer.

We studied the therapeutic effect of OK-432 combined with adoptive immunotherapy in 19 cases of liver metastases from breast cancer. Of the 14 patients who received intraarterial OK-432 injection and transfer of cultured lymphocytes, 9 responded to this therapy, whereas no patients responded to intravenous administration. The minimum cell number for a therapeutic response was 8 x 10(8) cells. Metastatic lesions other than those in the liver regressed after therapy in 4 patients. The serum carcinoembryonic antigen level paralleled the therapeutic effect. There were no severe side-effects accompanying this therapy. These results indicate that intraarterial adoptive immunotherapy combined with OK-432 is effective as a new therapeutic approach against liver metastases from breast cancer.

Adult↗