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Biomedical subjects

A Ohkubo

Publications and source records attributed to A Ohkubo.

At least 37 records · Page 2Linked to original sources

[Granulocyte-colony stimulating factor and clinical laboratory].

The effects of granulocyte-colony stimulating factor (G-CSF) on the white blood cell count and the morphology of peripheral white blood cells were studied during the period of leukocytopenia following chemotherapy in patients with non-hematological malignant tumors (solid tumor group) and those with lymphatic hematological diseases (lymphocytic malignancy group). Cell surface markers were also analyzed to evaluate the usefulness of G-CSF in peripheral stem cell transplantation. After G-CSF administration, the white blood cell count showed remarkable changes beyond the physiologic range; the increase and decrease were 5.0 times and 0.3 times previous values, respectively, in the solid tumor group, and 11.0 times and 0.2 times, respectively, in the lymphocytic malignancy group. Concerning the morphology of peripheral white blood cells, immature leukocytes increased markedly to 133.0 times the previous value. The frequency of immature cells with morphological abnormalities was 0.5 percent in the solid tumor group, but 1.6 percent in the lymphocytic malignancy group. These findings suggest that administering G-CSF affects precision control and microscopic findings in routine laboratory blood tests and that information about whether the patient was treated with G-CSF should be obtained from the clinical staff. A cell surface marker, c-kit, was analyzed to clarify whether stem cells useful for peripheral stem cell transplantation appear in peripheral blood after administration of G-CSF. The incidence of c-kit-positive cells was at 1-10% in 3 of 5 patients in the solid tumor group administered G-CSF and in all 6 patients in the lymphocytic malignancy group.(ABSTRACT TRUNCATED AT 250 WORDS)

Adolescent↗

[Detection of anticardiolipin antibody using the EIA kit prepared to eliminate interference of serum cofactor].

We studied sera from patients with various disorders including collagen diseases, Buerger's disease, deep vein thrombosis, repeated abortions and idiopathic thrombocytopenic purpura in order to measure anticardiolipin antibody (aCL) titers. In our assay system, we can detect aCL against bovine cardiolipin and the complex of cardiolipin and beta 2-glycoprotein I (beta 2-GPI). This was done with simultaneous assays using blank wells and bovine cardiolipin coated wells in the EIA method in which both wells were also coated with bovine serum albumin possibly containing beta 2-GPI, then aCL titers was given by subtraction of O.D. values of blank wells from those of cardiolipin coated wells. When the aCL was quantified by anti human immunoglobulin antibodies, collagen diseases showed positive aCL in 15 (11.5%) out of 130 sera with positive anti ENA antibodies, 3 (11.1%) out of 27 sera with positive anti DNA antibodies and 3 (5.2%) out of 58 sera with other positive ANA sera. The other hand, we found positive aCL in 2 (4.9%) out of 41 sera from patients with Buerger's disease, 4 (36.4%) out of 11 sera from patients with deep vein thrombosis, 1 (7.7%) out of 13 sera from patients with repeated abortions and 6 (14.6%) out of 41 sera from patients with idiopathic thrombocytopenic purpura. Twenty one (61.8%) out of these aCL positive sera had also positive IgG-aCL for the assay using anti human IgG antibody. Compared to previous reports, we thought, low incidence of aCL in our study was due to exclusion of anti beta 2-GPI antibody in our assay system.

Antibodies, Anticardiolipin↗

Identification of three types of PDGF-A chain gene transcripts in rabbit vascular smooth muscle and their regulated expression during development and by angiotensin II.

PDGF-like peptides secreted from smooth muscles have been suggested to be responsible for the smooth muscle growth. In order to elucidate the nature of PDGF-like molecules expressed in vascular smooth muscles, we have isolated and characterized cDNA clones for PDGF-A chain from a rabbit embryonic aorta cDNA library. One of the cDNA clones was found to encode a novel PDGF-A chain, named PDGF-A3 in this report. PDGF-A3 arises from a single PDGF-A chain gene by alternative RNA splicing and differs from the sequences of previously reported endothelial- or the glioma-type transcripts by a 110 bp insertion. Expression of PDGF-A3 mRNA was selectively induced by Angiotensin II in the smooth muscle cell in vitro. Total PDGF-A mRNA is most enriched in embryonic aortas, but its expression is down-regulated with vascular development. PDGF-A mRNA is markedly increased in primary-cultured smooth muscle cells during the log-phase growth. Our results suggest that autocrine production of PDGF-A chains from the smooth muscle cell may play a role in early vascular development and in Angiotensin II-induced smooth muscle cell proliferation.

Aging↗

Various factors associated with the manifestation of influenza-like illness.

A survey enrolling 814 schoolchildren was conducted immediately after a peak epidemic in the 1988-1989 influenza season and then a case-control study for influenza-like illness (ILI) was carried out using information concerning illness onset and usual lifestyle. Based on the analysis of the correlations among symptoms and actions taken due to symptoms, cases were defined as those with fever greater than or equal to 38 degrees C and subsequent absenteeism and medical consultation during peak epidemic; within this, mild-ILI (MILI) was defined as fever greater than or equal to 38 degrees C and less than 39 degrees C, and severe-ILI (SILI) as fever greater than or equal to 39 degrees C. Controls were defined as those with no symptoms (NS) during that period. Adjusted odds ratios (OR) were obtained through an unconditional logistic regression model between the MILI (80) or SILI (48) and NS (196) groups from among respondents (803). For MILI, increased risks were observed for easily-inflamed tonsils (OR = 3.0, 95% CI: 1.7-5.4); and larger family size (1.9, 1.1-3.4); with decreased risks for higher school grades (0.4, 0.2-0.9); frequent intake of vegetables or fruits other than green/yellow vegetables (0.5, 0.3-1.0); and larger room space per capita (0.4, 0.2-0.9). For SILI, there were increased risks for easily-inflamed tonsils (3.8, 1.8-8.1) and history of doctor-diagnosed asthma (2.9, 1.2-6.7); and decreased risks for higher grades (0.2, 0.1-0.6), frequent intake of milk or dairy products (0.3, 0.1-0.6) and vaccination (0.3, 0.1-0.8).

Case-Control Studies↗

[Recent advances in laboratory tests for liver diseases].

Many tests for hepatitis C virus (HCV) infection have been developed and have proved useful for prevention of post-blood transfusion hepatitis C. However, there are at least 4 genotypes of HCV and the predominant type is different among countries. None of the tests using antigens from one genotype are sensitive in detecting the antibodies against another genotype. More sensitive tests using a more stable part of the HCV RNA sequences such as 5'-noncoding region must be developed for clinical use. Automated PCR methods and DNA sandwich hybridization methods using branched DNA amplification multimers may be candidates. Recently a hepatocyte growth factor test has been developed in Japan. Multicenter trials of this test reveal that it is useful for assessment of acute severe hepatitis. Tests for collagen type IV, fibronectin receptor, and prolyl hydroxylase have been reported useful for assessment of liver fibrosis. However, serum prolyl hydroxylase is prone to increase in response to hepatocellular damage as well as fibrotic processes. Enzymatic methods for determination of branched amino acids and tyrosine have been developed. The molar ratio of branched amino acids to tyrosine seems to have same pathophysiological meaning as the ratio of branched amino acids to aromatic amino acids (Fischer ratio) in assessment of liver cirrhosis. Lidocaine test is reported to be useful for predicting survival of transplanted liver and also assessing the function of the cirrhotic liver. Profiles of alpha-fetoprotein subfractions based on lectin-reactivity and galactosyl transferase II isoenzyme have been reported to be useful for detecting hepatocellular carcinoma but this remains to be proved.

Clinical Laboratory Techniques↗

[Effects of antibiotics on the dissociation of platelet adherence in pseudothrombocytopenia].

To elucidate the effects of antibiotics on platelet adherence phenomena, we examined dissociation of platelet adherence in EDTA-dependent pseudothrombocytopenia by adding Kanamycin or Colimycin. We found the rapid dissociation of platelet adherence caused by addition of Kanamycin (20 mg) or Colimycin (20 mg) in 1 ml of EDTA-anticoagulated blood from patients with EDTA-dependent pseudothrombocytopenia. This dissociation was most remarkable in the sample treated within 30 minutes after venipuncture. When the dissociation occurred, no adherent platelet was found on microscopical examinations. Complete blood cell counts and their histogram patterns after dissociation were almost same as those just after venipuncture. The dissociation occurred as soon as the antibiotics were added and remained fairly long time. The dissociation decreased in proportion to the periods of time between venipuncture and addition of antibiotics. The dissociation was caused by many other antibiotics. However, severer damage were found morphologically by adding most of them in both histogram patterns and microscopical examinations than those of aminoglycoside or peptide antibiotics.

Adolescent↗

Analysis for cerebrospinal fluid proteins by sodium dodecyl sulfate-polyacrylamide gel electrophoresis.

Cerebrospinal fluid (CSF) proteins with molecular masses of < 150,000 Da were identified by immunoblotting after two kinds of nonreducing sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). With PAGE 1 (17-27% gradient gel), CSF proteins were clearly separated into seven to nine bands with molecular masses of 3000-67,000 Da; seven bands were identified as beta 2-microglobulin, lysozyme, prealbumin, free kappa and lambda chain, apolipoprotein A-I, glycoproteins, and albumin by immunoblotting. With PAGE 2 (10-20% gradient gel), proteins were clearly separated into 11-16 bands with molecular masses of 15,000-150,000 Da; 11 were identified as prealbumin, free kappa and lambda chain, apolipoprotein A-I, glycoproteins, albumin, alpha 1-antitrypsin, transferrin (separated into two bands), immunoglobulin fragments, haptoglobin, and IgG. We analyzed CSF samples collected from 81 patients with cerebrospinal signs by these SDS-PAGE methods and observed prominent bands in some cases.

Adolescent↗

Endoscopic ultrasonography findings in acute gastric anisakiasis.

To clarify the components of the thickened gastric wall in acute gastric anisakiasis, we evaluated endoscopic ultrasonography (EUS) findings of acute gastric anisakiasis. Ten patients with acute gastric anisakiasis underwent endoscopic ultrasonography immediately after anisakiasis was diagnosed by endoscopy. In the acute phase of gastric anisakiasis, endoscopic ultrasonography (EUS) detected a thickening of the gastric wall made of mainly thickened third layer with low echoic changes. When EUS was performed 7 to 11 days later in seven patients, the EUS findings had normalized. We made clear EUS findings in acute gastritis due to gastric anisakiasis. Consequently, we found, by EUS, that the main inflammatory lesion in anisakiasis was in the submucosal layer of gastric wall.

Acute Disease↗

[The clinical usefulness of the molar ratio of branched-chain amino acids to tyrosine (BTR) in discriminating stage of chronic liver diseases].

We determined the molar ratio of branched-chain amino acids to tyrosine (BTR) in plasma and in serum by enzymatic method and compared it with Fischer ratio (the molar ratio of branched-chain amino acids to tyrosine and phenylalanine) in plasma obtained by conventional HPLC method. BTR in plasma and in serum was well correlated with plasma Fischer ratio. The normal range (mean +/- 2SD) of BTR was determined to be 4.41-10.05 in 210 normal subjects. In addition, we investigated the distribution of BTR values in patients with various liver diseases. BTR value decreased according to the severity of liver disease. We evaluated the clinical usefulness of BTR in patients with chronic liver diseases by cumulative distribution analysis (CDA) graph and receiver operating characteristic curve (ROC) analysis. The area under the curve for BTR analyzed by ROC for CH versus LC.HCC group was the highest (86.3%) of any for various concurrently-measured liver function tests, and was significantly higher than AST/ALT, ALT, AST, gamma-GT (each, p less than 0.001) and ALB (p less than 0.05). These diagnostic results showed that BTR is a superior indicator in discriminating between liver cirrhosis and chronic hepatitis.

Adult↗

Determination of proteins in urine by high-performance liquid chromatography with spectrophotometric detection using a pyrogallol red-molybdate complex.

A high-performance liquid chromatographic method with spectrophotometric detection was developed for the determination of proteins in urine. The proteins were separated on an anion-exchange column and eluted with a Tris-HCl buffer with a gradient of sodium chloride concentration and pH. The separated proteins were mixed with a pyrogallol red-molybdate complex reagent and determined spectrophotometrically. Urinary proteins were well separated without desalting the urine. The reproducibility was satisfactory.

Chromatography, High Pressure Liquid↗

cDNA cloning of a myosin heavy chain isoform in embryonic smooth muscle and its expression during vascular development and in arteriosclerosis.

Adult rabbit smooth muscles contain two types of myosin heavy chain (MHC) isoforms, SM1 and SM2 which are generated through alternative RNA splicing from a single gene (Nagai, R., Kuro-o, M., Babij, P. & Periasamy, M. (1989) J. Biol. Chem. 264, 9734-9737). We previously reported that the expression of SM1 and SM2 during vascular development is differentially regulated at the level of RNA splicing, whereby SM1 is constitutively expressed from early development but SM2 appear after birth (Kuro-o, M., Nagai, R., Tsuchimochi, H., Katoh, H., Yazaki, Y., Ohkubo, A. & Takaku, F. (1989) J. Biol. Chem. 264, 18272-18275). We also demonstrated that embryonic vascular smooth muscles contain a third type of MHC isoform, referred to as SMemb in this report, which comigrates on sodium dodecyl sulfate-polyacrylamide gel electrophoresis with SM2. In the present study we have isolated and characterized a cDNA clone (FSMHC34) for SMemb. FSMHC34 encodes the light meromyosin region including the carboxyl terminus and showed 70% amino acid sequence identity with SM1 or SM2. SMemb is a nonmuscle-type MHC and identical with brain MHC, but clearly distinct from 196-kDa nonmuscle MHC in cultured smooth muscle cells. The expression of SMemb was predominant in embryonic and perinatal aortas, but down-regulated with vascular development. Interestingly SMemb was reexpressed in proliferating smooth muscle cells of arteriosclerotic neointimas. These results suggest that smooth muscle proliferation is coupled to the expression of SMemb and that dedifferentiation of smooth muscles toward the embryonic phenotype is involved in the mechanisms underlying atherosclerosis.

Aging↗

Two-step hard acid deprotection/cleavage procedure for solid phase peptide synthesis.

A new two-step deprotection/cleavage procedure for t-butoxycarbonyl (Boc) based solid phase peptide synthesis is reported. First the protective groups are removed from 4-(oxymethyl)-phenylacetamidomethyl (PAM) resin attached peptide with the weak hard acid, trimethylsilyl bromide-thioanisole/trifluoroacetic acid (TFA). In the second step, the peptide is cleaved from the resin with a stronger hard acid such as trimethylsilyl trifluoromethanesulfonate in TFA or with HF. The method is also shown to deformylate Nin-formyltryptophan moiety efficiently. The usefulness of this procedure for practical solid phase peptide synthesis is demonstrated by comparison with other deprotection methods in the synthesis of urotensin II and human endothelin.

Acetamides↗

Preoperative evaluation of gastric cancer by endoscopic ultrasound.

The preoperative use of endoscopic ultrasound was evaluated in 74 patients with confirmed gastric cancer. It was used in diagnosing the depth of invasion in the gastric wall, the infiltration to the adjacent organs, and the involvement of the perigastric lymph nodes. Results were compared with histological findings in resected specimens. Accuracy in staging gastric cancer using the T grade of the 1987 TNM system was 81.1% (60 of 74 patients). Endoscopic ultrasound provided excellent results compared with computed tomography and conventional ultrasound, particularly in evaluating perigastric lymph node metastasis and direct infiltration to the adjacent organs. The success rate in detecting lymph node metastasis was 50% (11 of 22 patients); the accuracy in diagnosing direct infiltration to the adjacent organs was 60% (three of five patients). This technique is useful in diagnosing malignant invasion and lymph node metastasis of gastric carcinomas but requires further refinement for use in diagnosing the disease itself. Its preoperative use is recommended for establishing surgical and other treatment plans, as well as in predicting the prognosis of gastric cancer.

Gastroscopy↗

[Cytoplasmic staining pattern characteristic for anti Jo-1 antibody in HEp-2 cells].

We wished to confirm the staining pattern of HEp-2 cells by the anti Jo-1 antibody, because we found antibodies in serum with positive anti Jo-1 antibody which showed either a fibrilar cytoplasmic staining or a nuclear speckled staining pattern in indirect immuno fluorescence+ examinations using HEp-2 cells. Sera available from eight patients with PM DM (polymyositis and/or dermatomyositis) showed positive anti Jo-1 antibody in the double immuno-diffusion technique but had various staining patterns of HEp-2 cells in the immunofluorescent examination. We examined these eight sera with the immuno-blotting method utilizing whole cell extract of HeLa cells, and found the 50 kDa band from all sera tested, to which Jo-1 antigen had been reported to move. We eluted the antibody which formed the 50 kDa band from the nitrocellulose membrane and applied it on HEp-2 cells. This maneuver gave us the fine granular cytoplasmic staining of anti Jo-1 antibody on HEp-2 cells. We therefore concluded that the anti Jo-1 antibody should have cytoplasmic staining on HEp-2 cells although observers might miss it due to other types of associated antinuclear antibodies.

Antibodies, Antinuclear↗

[Magnetoencephalographic studies on spike foci using a 37-channel biomagnetometer system].

We report the results of a clinical trial of Magnetoencephalography (MEG) on spike foci in patients with epilepsy, which was performed from December 1990 to June 1991 at The University of Tokyo Hospital. Fifty patients with focal epilepsy; 26 primary epilepsy, 24 secondary epilepsy (7 brain tumor, 4 arteriovenous malformation, 4 encephalitis, 3 porencephaly, 2 arachnoid cyst, 1 brain abscess, 1 hemimegaloencephaly, 1 Lance-Adams syndrome, 1 hygroma), and ten normal subjects were enrolled in this study. MEG data were recorded using a 37-channel biomagnetometer system SMI-1001 (BTi Magnes, Biomagnetic Technologies, Inc., San Diego). A simultaneous 19-channel EEG recording with linked-ear reference was also obtained. The overall study was completed safely and none of the normal subjects showed abnormal paroxysmal MEG activity. Two patients showed interictal EEG spikes which would not have been noticed without first noting the presence of corresponding prominent MEG spikes. On the whole, the MEG signal seemed to have a wider frequency bandwidth than EEG. In most cases, the source localization predicted by MEG corresponded well with the EEG findings. The relative accuracy of MEG spike source localization was estimated to be within a cubic centimeter from the cases which showed tightly clustered localization of individual spikes. High-pass filtering reduced interference by superimposed slow wave activity, thereby improving the localization of spike sources. These results demonstrate that 37-channel biomagnetometer system could be a useful tool for analyzing epileptic spike sources.

Adolescent↗