Evaluation of coculture aggregation with TT2 cells for production of germline chimera.
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Animals↗
Biomedical subjects
J Ishida
Publications and source records attributed to J Ishida.
Angiotensinogen-deficient mice with hypotension.
The renin-angiotensin system is an enzymatic cascade that produces a potent vasoconstrictor octapeptide angiotensin II, through its physiologically inactive intermediate decapeptide angiotensin I, from their precursor angiotensinogen. In the present study, we generated angiotensinogen-deficient mice by homologous recombination in mouse embryonic stem cells. These mice do not produce angiotensinogen in the liver, resulting in the complete loss of plasma immunoreactive angiotensin I. The systolic blood pressure of the homozygous mutant mice was 66.9 +/- 4.1 mm Hg, significantly lower than that of wild-type mice (100.4 +/- 4.4 mm Hg). This profound hypotension in angiotensinogen-deficient mice demonstrates an indispensable role for the renin-angiotensin system in maintaining blood pressure.
Angiotensin I↗
Simultaneous high-performance liquid chromatographic determination of 6 beta-hydroxycortisol and cortisol in urine with fluorescence detection and its application for estimating hepatic drug-metabolizing enzyme induction.
A simple and sensitive high-performance liquid chromatographic method is described for the simultaneous determination of 6 beta-hydroxycortisol (6 beta-OHF) and cortisol (F) in urine. Urine (1 ml) containing fludrocortisone as the internal standard is extracted with ethyl acetate. The extract is washed successively with sodium hydroxide solution and water, and subsequently dried under a stream of nitrogen. The residue is redissolved in methanol. The 6 beta-OHF, F and fludrocortisone in the methanol solution are oxidized by cupric acetate and the resulting glyoxal compounds are converted into fluorescent derivatives with 1,2-diamino-4,5-methylenedioxybenzene (DMB). The DMB derivatives of the corticosteroids are separated within 70 min on a reversed-phase column, L-Column ODS, using stepwise elution with methanol-acetonitrile-0.5 M ammonium acetate and detected fluorimetrically at 350 nm (excitation) and 390 nm (emission). The lower limits of detection for 6 beta-OHF and F are 1.8 pmol (680 pg) and 2.4 pmol (950 pg)/ml urine (0.6 pmol and 0.8 pmol/100 microliters injection volume), respectively, at a signal-to-noise ratio of 3. This method can be applied to the determination of urinary 6 beta-OHF, and the ratio of 6 beta-OHF to F in humans and in rhesus monkeys treated orally with phenobarbital as a hepatic drug-metabolizing enzyme inducer.
Administration, Oral↗
Benign prostatic hyperplasia: value of MR imaging for determining histologic type.
PURPOSE: To assess the value of magnetic resonance (MR) imaging for determining the histologic type of benign prostatic hyperplasia (BPH). MATERIALS AND METHODS: Unenhanced T1- and T2-weighted images and gadolinium-enhanced T1-weighted images were obtained in the transverse plane at 1.5 T in 33 patients with BPH. Hyperphasia was classified as stromal or nonstromal. Nonstromal hyperplasia was diagnosed if (a) the nodules in the inner gland had heterogeneous high signal intensity on T2-weighted images and peripheral enhancement on gadolinium-enhanced images, (b) a surgical capsule was present, or (c) the inner gland volume to total volume ratio was greater than 0.75. RESULTS: A correct diagnosis was made with MR imaging in 23 of 24 patients with nonstromal hyperplasia and eight of nine with stromal hyperplasia (94% accuracy). In nonstromal hyperplasia, nodular lesions were seen in 20 patients and a surgical capsule was seen in 19. Eighteen patients had a transition zone ratio greater than 0.75. CONCLUSION: MR imaging appears to be useful for choosing the type of pharmacotherapy performed because accurate histologic assessment is possible.
Aged↗
[CT evaluation of the pelvic cavity after total cystectomy].
CT scans were performed 40 times in 28 postcystectomy patients to identify the type and location of tumor recurrence. Sixteen patients showed tumor recurrence. Patients without recurrence showed a complete symmetric pelvis and a thin string that looked like an operation scar. Cutaneous ureterostomy showed small tube-like structures, and ileal conduit looked like herniation of the ileum. Cases with recurrent tumors were divided into 3 patterns, 1) pelvic abscess recurrence, 2) anterior abdominal wall thickening, 3) pelvic lateral wall thickening, and combinations of these types. The abscess-recurrence type often showed air within the mass or a fuzzy contour that could not be differentiated from true inflammatory abscess. Anterior abdominal wall thickening was usually accompanied with abscess-type recurrence and appeared as thickening of the operation scar. Lateral wall thickening was usually seen at the site of the obturator internus muscle or obturator node. Patients without recurrence had no complaints except for two who had ileus. All patients with recurrence except one had complaints of pain, abnormal secretion, or a palpable mass. Because all patients with recurrence already had a relatively large tumor at the time of CT, routine follow-up CT is recommended even if the patient has no complaints.
Carcinoma, Transitional Cell↗
Comparative metabolism and structure of BCKD-E2 in primary biliary cirrhosis.
The identification and cloning of the mitochondrial autoantigens in primary biliary cirrhosis (PBC) have provided new clues in disease pathogenesis. The two major autoantigens are the E2 subunits of pyruvate dehydrogenase and branched-chain ketoacid dehydrogenase (BCKD). Interestingly, one of these complexes, BCKD-E2, is already well known to clinical medicine based on its association with genetic mutations in maple syrup urine disease (MSUD). Patients with this disease have an inability to metabolize branched-chain amino acids. In the present study, we have taken advantage of the known sequence of BCKD-E2 from normal humans, and addressed the issue of whether there is an altered autoantigen sequence in hepatocytes of individuals with primary biliary cirrhosis. In particular, we examined both the leader sequence and the B-cell immunodominant epitope, the lipoic acid domain. In addition, because patients with PBC have autoantibodies to the BCKD-E2 complex, we have quantitated plasma levels of alpha-ketoacids potentially affected in maple syrup urine disease. These include pyruvic acid (PY), phenylpyruvic acid (PP), alpha-ketoisocaproic acid (KIC) alpha-ketoisovalerate (KIV) and alpha-keto-beta-methylvaleric acid (KMV). The levels of these alpha-ketoacids were compared in patients with primary sclerosing cholangitis and normal volunteers. The sequence of BCKD-E2 obtained from PBC hepatocytes showed homology with normal BCKD. Further studies of autoantigen structure and sequence are clearly indicated, including those involved in mitochondrial transport and localization. Finally, we noted a statistically significant increase in all plasma alpha-ketoacids except alpha-keto-beta-methylvaleric acid in PBC patients.(ABSTRACT TRUNCATED AT 250 WORDS)
3-Methyl-2-Oxobutanoate Dehydrogenase (Lipoamide)↗
Malignant anorectal melanoma: usefulness of fat saturation MR imaging.
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Aged↗
High-performance liquid chromatographic determination of 5-hydroxyindoles by post-column fluorescence derivatization.
A selective and sensitive high-performance liquid chromatographic method with post-column fluorescence detection has been developed for the determination of 5-hydroxyindoles. Five biogenic 5-hydroxyindoles (5-hydroxytryptophan, 5-hydroxytryptamine, 5-hydroxyindol-3-ylacetic acid, 5-hydroxytryptophol and N-acetyl-5-hydroxytryptamine) were separated by isocratic elution on a reversed-phase column, TSKgel ODS-80Tm, and then converted into fluorescent derivatives by post-column reaction with benzylamine and potassium hexacyanoferrate(III) reagents. The detection limits (signal-to-noise ratio = 3) for the indoles were in the range 140-470 fmol per injection volume (100 microliters). The method was applied to the determination of 5-hydroxyindol-3-ylacetic acid in human urine, with direct sample injection.
Adult↗
Serotonin and 5-hydroxyindole-3-acetic acid in human plasma and rat brain determined by liquid chromatography with post-column derivatization and fluorescence detection.
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Animals↗
High-performance liquid chromatographic determination of 3 alpha,5 beta-tetrahydroaldosterone in human urine with chemiluminescence detection.
A sensitive method for the determination of 3 alpha,5 beta-tetrahydroaldosterone (THALD) in human urine is described. The method uses high-performance liquid chromatography with chemiluminescence detection. Urinary THALD, released by enzyme hydrolysis, is isolated and concentrated using a Sephadex G-25M column and Bond-Elut C1 cartridges, and then oxidized by copper(II) acetate to form the corresponding glyoxal derivative. The glyoxal derivative is converted into the chemiluminescent quinoxaline by reaction with 4,5-diaminophthalhydrazide. The chemiluminescent quinoxaline is separated within 50 min on a reversed-phase column (TSKgel ODS-120T) with isocratic elution, followed by chemiluminescence detection; the chemiluminescence is produced by the reaction of the quinoxaline with hydrogen peroxide in the presence of potassium hexacyanoferrate(III) in alkaline solution. The detection limit for THALD is 0.6 pmol (220 pg) ml-1 in urine [1.5 fmol (0.53 pg) per 20 microliters injection] at a signal-to-noise ratio of 3. This method permits the sensitive and precise determination of THALD in human urine (50 microliters) from normal subjects and a patient with primary aldosteronism.
Aldosterone↗
High-performance liquid chromatographic determination of fatty acid binding proteins in rat liver with fluorescence detection.
A highly sensitive, simple and reproducible method for the quantitative determination of fatty acid binding protein in rat liver by post-column high-performance liquid chromatography with fluorescence detection is presented. Fatty acid binding protein in rat liver cytosols is separated by gel-permeation column chromatography, which is followed by fluorescence detection. The detection makes use of the fluorescence enhancement observed when a fluorescent fatty acid probe, dansylundecanoic acid, binds to fatty acid binding protein. The method is rapid, simple to perform and highly sensitive. The method was applied to the determination of fatty acid binding protein in liver from control and hypolipidaemic drug treated rats.
Animals↗
High-performance liquid chromatographic determination of 1,2,3,4-tetrahydroisoquinoline in rat brain with fluorescence detection.
A high-performance liquid chromatographic method for the fluorometric determination of 1,2,3,4-tetrahydroisoquinoline in rat brain is described. 1,2,3,4-Tetrahydroisoquinoline and 4-phenylpiperidine (internal standard) are isolated by liquid-liquid extraction, and then converted into the corresponding fluorescent derivatives with 3,4-dihydro-6,7-dimethoxy-4-methyl-3-oxoquinoxaline-2-carbonyl chloride, a fluorescence derivatization reagent for amines. The derivatives are separated within 60 min on a reversed-phase column, TSK gel ODS-120T, with isocratic elution, and detected fluorometrically. The detection limit of 1,2,3,4-tetrahydroisoquinoline is 1.0 pmol/g in rat brain (S/N = 3).
Animals↗
Spectrofluorimetric determination of 5-hydroxyindoles with benzylamine or 3,4-dimethoxybenzylamine as a selective fluorogenic reagent.
A spectrofluorimetric method has been developed for the sensitive and selective determination of 5-hydroxyindoles; the method is based on the reaction of 5-hydroxyindoles in a weakly alkaline solution (pH 9.0) with aromatic methylamines in the presence of potassium hexacyanoferrate(III) and dimethyl sulphoxide; the compounds produced fluoresce intensely in an alkaline solution (pH 11-12). Of the eight aromatic methylamines tested, benzylamine and 3,4-dimethoxybenzylamine were found to be the most favourable fluorogenic reagents in terms of sensitivity and reactivity. The methods with benzylamine and 3,4-dimethoxybenzylamine permit the determination of 5-hydroxyindoles at concentrations as low as 22-72 pmol ml-1 and 1.0-2.4 nmol ml-1, respectively.
Benzylamines↗
Nasal tip treatment by lengthening of the short columella in bilateral cleft lip sequelae.
The mobilization of two muco-chondrocutaneous flaps obtained by two parallel incisions along the nasal dorsum may elevate and elongate the columella in patients with bilateral cleft lips. The nose tip is also modulated by a triangular tissue resection right above the alar segment in both sides. Thirty-four patients have been treated with this method.
Adolescent↗
Determination of alpha-keto acids including phenylpyruvic acid in human plasma by high-performance liquid chromatography with chemiluminescence detection.
A highly sensitive method for the determination of alpha-keto acids including phenylpyruvic acid in human plasma is investigated. The method employs high-performance liquid chromatography with chemiluminescence detection. The acids and alpha-ketocaproic acid (internal standard) in human plasma are isolated by anion-exchange chromatography on a Toyopak DEAE cartridge, and then converted into the corresponding chemiluminescent derivatives with 4,5-diaminophthalhydrazide dihydrochloride, a chemiluminescence derivatization reagent for alpha-keto acids. The derivatives are separated within 50 min on a reversed-phase column, TSKgel ODS-120T, with isocratic elution, followed by chemiluminescence detection; the chemiluminescence is produced by the reaction of the derivatives with hydrogen peroxide in the presence of potassium hexacyanoferrate(III). The detection limits for the acids are in the range 9-92 pmol/ml in plasma (signal-to-noise ratio = 3). This sensitivity permits precise determination of several alpha-keto acids including phenylpyruvic acid, which cannot be determined by other HPLC methods, in 10 microliters of normal human plasma. The chemiluminescent product from phenylpyruvic acid was characterized as 3-benzyl-7,8-dihydropyridazino[4,5-g]quinoxaline-2,6,9(1H)-trione.
Adult↗
[Malignant ovarian tumor invasion to the uterus and bladder: assessment with magnetic resonance imaging].
Twenty-nine patients with ovarian malignant tumor were studied by means of magnetic resonance (MR) imaging. The findings were compared with surgical findings concerning tumor invasion to the bladder and to the uterus. The bladder invasion was evaluated with MRI and classified as follows: (1) Huge amount of fat, colon or small intestine between the tumor and the bladder. (2) Thin fat layer between the bladder and the tumor, (3) No fat layer, and the bladder was smoothly compressed by the tumor, (4) No fat layer with local deformity of the bladder caused by the tumor existed, or tumor protruded into the bladder. In group (4), 4 of 5 cases were proved to have bladder invasion. However in group (3), half of the 14 cases showed bladder invasion and the others showed no invasion. Criteria used in MRI analysis of uterine invasion was as follows: (1) Fat layer between the uterus and the tumor, (2) Fat layer between the tumor and the bladder, and the uterine contour was indistinct, (3) No fat layer and deformity of the uterus was apparent. In group (3), many cases showed uterine invasion, but 4 cases out of 12 cases did not. In the group (2), 3 out of 11 cases showed uterine invasion. The diagnostic ability of uterine invasion was not as accurate as in cases of the bladder invasion. The MRI sagittal plane can directly show the relationship between the uterus, the bladder and tumor, so that diagnosis of tumor invasion by MRI is superior to CT. However MRI could not differentiate between adhesion and invasion.
Adenocarcinoma↗
High-performance liquid chromatographic determination of beta-phenylethylamine in human plasma with fluorescence detection.
A simple and highly sensitive method for the determination of beta-phenylethylamine in human plasma is investigated. The method employs high-performance liquid chromatography with fluorescence detection. beta-Phenylethylamine and p-methylbenzylamine (internal standard) in human plasma are isolated by cation-exchange chromatography on a Toyopak SP cartridge and then converted into the corresponding fluorescent derivatives with 3,4-dihydro-6,7-dimethoxy-4-methyl-3-oxoquinoxaline-2-carbonyl chloride, a fluorescence derivatization reagent for amines. The derivatives are separated within 30 min on a reversed-phase column, TSK gel ODS-120T, with isocratic elution, and detected fluorometrically. The detection limit of beta-phenylethylamine is 0.3 pmol/ml in plasma (S/N = 3).
Benzylamines↗
The relative effects of collagen fiber orientation, porosity, density, and mineralization on bone strength.
This investigation determined the relative importance of collagen fiber orientation, porosity, density, and mineralization in determining the tensile strength of bovine cortical bone. Thirty-nine specimens were tested for failure stress and the values of eight histologic and compositional parameters: collagen fiber orientation, wet and dry apparent density, percent mineralization of the bone matrix, and several components of porosity (Haversian canals, Volkmann's canals, and plexiform vascular spaces). Linear regression analysis showed that collagen fiber orientation was consistently the single best predictor of strength. Mineralization of the bone matrix was generally a poor predictor of strength. Density and porosity ranked between these variables in importance. Multiple regression equations containing all significantly correlated variables achieved correlation coefficients of 0.607 for plexiform bone and 0.881 for osteonal bone. Also, separate analysis of plexiform and osteonal specimens showed that the latter group was weaker even though it was less porous, apparently because it had collagen fibers which were less longitudinally oriented. This study suggests it is feasible to develop better empirical formulae for the prediction of cortical bone strength than are currently available if a variety of variables is introduced. Additional data are needed to confirm these results.
Animals↗