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Biomedical subjects

E Ishikawa

Publications and source records attributed to E Ishikawa.

At least 91 records · Page 5Linked to original sources

Novel acyl-CoA:cholesterol acyltransferase inhibitors. Synthesis and biological activity of 3-quinolylurea derivatives.

A series of 3-quinolylurea derivatives (1) was synthesized and evaluated for acyl-CoA:cholesterol acyltransferase (ACAT) inhibitory activity. For in vitro studies, the most potent inhibitory activity was found in derivatives having substituents at the 6,7- or 6,8-positions and an ortho-substituted phenyl group at the 4-position of quinoline ring. The 2,4-difluorophenyl group appeared to be the optimum N'-substituent of the urea moiety. The IC50 values of compounds 52-54 and 59 were in the nanomolar order. Plasma cholesterol-lowering activity of compounds 50, 52, and 54 was observed at less than 1 mg/kg/day in cholesterol-fed rats. Compound 52 was also hypocholesterolemic in hamsters fed a diet without loading cholesterol.

Animals↗

Anti-HTLV-I IgG in urine detected by sensitive enzyme immunoassay (immune complex transfer enzyme immunoassay) using a synthetic peptide, Cys-env gp46(188-224), as antigen.

Antibody IgG to human T-cell leukemia virus type I (HTLV-I) in urine was detected by a sensitive enzyme immunoassay (immune complex transfer enzyme immunoassay) using a synthetic peptide, Cys-env gp46(188-224), as antigen, the sensitivity and specificity of which were 100 and 98.5%, respectively, using serum samples. Anti-HTLV-I IgG in urine was reacted simultaneously with 2,4-dinitrophenyl-bovine serum albumin-Cys-env gp46(188-224) conjugate and Cys-env gp46(188-224)-beta-D-galactosidase (Escherichia coli) conjugate. The complex formed, consisting of the three components, was trapped onto polystyrene balls coated with affinity-purified (anti-2,4-dinitrophenyl group) IgG, eluted with epsilon N-2,4-dinitrophenyl-L-lysine and transferred to polystyrene balls coated with affinity-purified (anti-human IgG gamma-chain) IgG. Finally, bound beta-D-galactosidase activity was assayed by fluorometry. Thirty-one urine samples from seropositive subjects and 100 urine samples from seronegative subjects were tested. The sensitivity and specificity were 87 and 100%, respectively, with unconcentrated urine samples and 94 and 100%, respectively, with approximately 10-fold concentrated urine samples. These results were superior to those by the conventional ELISA and gelatin particle agglutination test.

Adult↗

Use of microplates and fluororeader for ultrasensitive enzyme immunoassay (immune complex transfer enzyme immunoassay) of anti-HTLV-I IgG.

Previously, an ultrasensitive solid phase enzyme immunoassay (immune complex transfer enzyme immunoassay) was described to detect low levels of anti-HTLV-I IgG in serum below those detectable by conventional methods. In this method, polystyrene balls as solid phase were transferred from test tube to test tube with tweezers. This was not only tedious but also causative of false-positivity by carryover, unless tips of the tweezers were washed carefully after each transfer of polystyrene balls. Bound enzyme activities for many samples were measured one by one by fluorometry using a spectrofluorophotometer. As a result, the assay of many samples was difficult. In the present study, microplates and a fluororeader were used in place to test tubes and a spectrofluorophotometer. Polystyrene balls were transferred quickly and easily from well to well by placing a microplate upside down on that containing polystyrene balls, and turning the two plates together upside down. Tweezers were not used for transfer of polystyrene balls, minimizing the possibility of false-positivity. Fluorescence intensities of bound enzyme activities for 96 samples were measured within a minute by using a fluororeader. Thus, it became easy to test many samples, although the sensitivity was lowered to some extent.

Amino Acid Sequence↗

Thyroglobulin antibodies in type 1 diabetic patients and their relatives--measurement with highly sensitive assay.

Autoimmunity in Type 1 (insulin-dependent) diabetes mellitus was assessed by measuring thyroglobulin antibodies (TGA) using a highly sensitive enzyme immunoassay in 65 young patients with Type 1 diabetes mellitus, 83 healthy first-degree relatives of the patients, 37 healthy control subjects and 67 healthy parents of the control subjects. TGA were found in 78.5% (51/65) of patients and were significantly more frequent in patients than in control subjects (40.5%, 15/37; P < 0.01). The prevalence of TGA in patients showed no correlation with age at onset, duration of diabetes or sex. Among the first-degree relatives the prevalence of TGA was significantly increased in mothers of patients than in mothers of the control subjects (80.0% vs. 54.3%, P < 0.05), while not significantly between fathers of patients and fathers of control subjects or between siblings of patients and control subjects. Comparing the TGA levels of TGA-positive subjects, the TGA levels in patients, their parents and their siblings were significantly higher than those in the corresponding control subjects (P < 0.05, P < 0.05 and P < 0.01, respectively). In the present study we thus more clearly demonstrated autoimmune diathesis in patients with Type 1 diabetes and in their first-degree relatives.

Adolescent↗

Ossifying subcutaneous tumor with myofibroblastic differentiation: a variant of ossifying fibromyxoid tumor of soft parts?

Immunohistochemical, ultrastructural, and flow cytometric studies were performed on an ossifying soft-tissue tumor, presumed to be a variant of ossifying fibromyxoid tumor of soft parts, which was located in the subcutis of the left buttock of a 76 year old Japanese woman. Histologically, this was a benign-looking spindle, oval, or round cell lesion, having a fibrous capsule with a discontinuous rim of bone as seen in typical cases. However, this lesion was also characterized by a high degree of cellular proliferation in storiform and whorl arrangements, extensive ossification, osteoid and metaplastic bone formation and absence of myxoid features. In an immunohistochemical study using formalin-fixed, paraffin-embedded sections, many tumor cells expressed vimentin, S-100 protein, Leu-7, neuron specific enolase, and desmin. Ultrastructurally, this neoplasm consisted of fibroblast-like cells and myofibroblast-like cells. This tumor had an aneuploid DNA content. No recurrence has been observed for 16 months. These results suggest that the neoplastic cells may show the phenotypic expressions of myofibroblast and also osteogenic differentiation.

Aged↗

Detection of anti-human immunodeficiency virus type 1 (HIV-1) immunoglobulin G in urine by an ultrasensitive enzyme immunoassay (immune complex transfer enzyme immunoassay) with recombinant reverse transcriptase as an antigen.

Anti-human immunodeficiency virus type 1 immunoglobulin G in urine was detected by an immunoassay with reverse transcriptase as the antigen and beta-D-galactosidase as the label; this immunoassay was 30-fold more sensitive than the previous immunoassay with peroxidase as the label. The sensitivity and specificity were both 100%. The lowest signal for asymptomatic carriers was 20-fold higher than the highest signal for seronegative subjects.

Adolescent↗

A novel hypothyroid 'growth-retarded' mouse derived from Snell's dwarf mouse.

This paper describes a novel mutant mouse that has been spontaneously derived from the Snell's dwarf (DW/J) mouse. It was named the 'growth-retarded mouse' because of a characteristic growth pause followed by the delayed onset of pubertal growth. The onset of the increase in pituitary GH content that normally occurs concomitant with pubertal growth was also delayed in the growth-retarded mice. The serum concentration of thyroxine was very low in these mice from the neonatal period through adulthood, and a supplement of tri-iodothyronine was effective in shortening the growth pause and commencing the suppressed pubertal growth. Histological and immunohistochemical studies revealed that the anterior pituitary gland of the growth-retarded mouse contains clustered unusual chromophobic cells which are not reactive to various antisera against anterior pituitary hormones and the gland becomes enlarged with age. Breeding data indicated that these characteristics of the mice show an autosomal recessive inheritance and the gene responsible was designated as 'grm'. Partial linkage analysis utilizing microsatellite polymorphism demonstrated that the grm gene does not identify with the lit or hyt genes. Based on comparison of the hormonal status and growth pattern between growth-retarded, dwarf and normal mice, we have suggested the existence of a mutual interaction, possibly positive feedback regulation, between the pituitary and thyroid glands, that develops or matures the hormonal network which is responsible for rapid somatic growth and metabolic changes at puberty in mice.

Animals↗

A patient with primary hypoparathyroidism developing hypercalcemia associated with adult T-cell leukemia/lymphoma.

A 38-year-old woman was admitted to our hospital with symptoms and signs of hypocalcemia in 1977 and a diagnosis of primary hypoparathyroidism was made with a positive Ellsworth Howard test. She was then lost to follow up until 1992 when she returned this time with symptoms and signs of hypercalcemia. An inguinal lymph node was biopsied showing non-Hodgkin's lymphoma, diffuse pleomorphic type and monoclonal integration of proviral human T-cell lymphotropic virus-1 DNA was detected in lymph node cells indicating ATLL. Serum parathyroid hormone-related peptide (PTHrP) was slightly elevated and the tumor cells were positively stained with anti-PTHrP serum. Combination chemotherapy with vincristine, adriamycin, cyclophosphamide and prednisolone was given to the patient with disappearance of the lymphadenopathy and subsequent normalization of PTHrP levels. Interestingly, the signs and symptoms of hypocalcemia reappeared after the treatment requiring replacement therapy with calcium and vitamin D.

Adult↗

[A case of infrarenal left inferior vena cava with macrohematuria after contusion in the lumber region].

A 33-year-old woman was admitted with macrohematuria after contusion in the left lumber region. The excretory urogram showed no abnormal findings, but color doppler and RI-angiography revealed infrarenal left-sided inferior vena cava (left IVC). The pressure in the dilated left renal vein was higher than that in the right one. Renal contusion in addition to the congestion of the left renal vein due to left IVC was thought to be the cause of hematuria in this case. Hematuria disappeared after retrograde instillation of 0.2% AgNO3 water solution into the left renal pelvis. Left IVC rarely presents clinical symptoms, and is usually found by chance on CT scan or ultrasonography during examinations for upper abdominal organs. Our case is the 69th one of left IVC in Japan, and belongs to Type 2' according to our classification of left IVC.

Adult↗

Soft tissue vascular tumors. A flow cytometric DNA analysis.

BACKGROUND: The clinical behavior of soft tissue vascular tumors is difficult to predict on histologic grounds alone. METHODS: To assess the usefulness of DNA flow cytometry in predicting the biologic behavior of these tumors, the authors studied 51 soft tissue vascular tumors by DNA flow cytometry of paraffin-embedded tissue. RESULTS: All 20 capillary hemangiomas, one epithelioid hemangioma, two spindle cell hemangioendotheliomas, and two benign hemangiopericytomas had a diploid DNA content. Of the 20 patients with angiosarcomas, their ages ranged from 9-80 years (mean, 44.0 years), and the mean follow-up period was 16.5 months. Ten tumors (including two postmastectomy angiosarcomas) were cutaneous, six were deep soft tissues, and four were from other sites. These tumors were classified histologically into three types: sinusoidal (n = 10), capillary (n = 7), and mixed type (n = 3). Thirteen of the angiosarcomas (65%) were diploid, and seven (35%) were aneuploid. Three patients with angiosarcoma had lung metastases at initial diagnosis, and seven had them later in the course. Nine patients died of their disease within a period of 18 months after the initial diagnosis. All five patients with malignant hemangiopericytoma died of their disease. Three of these lesions had a diploid DNA content, and two had an aneuploid content. Although it was found that all benign and intermediate-grade malignant tumors were diploid, there were no significant correlations among histologic type, DNA ploidy, and clinical outcome in angiosarcomas and malignant hemangiopericytomas. CONCLUSIONS: Based on this study, DNA analysis has limited value in predicting the biologic behavior of malignant vascular tumors of the soft tissue.

Adolescent↗

Measurement of anti-thyroglobulin IgG in urine of patients with autoimmune thyroid diseases by sensitive enzyme immunoassay (immune complex transfer enzyme immunoassay).

Anti-thyroglobulin IgG in urine of patients with Graves' disease and chronic thyroiditis and healthy subjects was measured by a sensitive enzyme immunoassay (immune complex transfer enzyme immunoassay). Anti-thyroglobulin IgG in dialyzed urine was reacted simultaneously with 2,4-dinitrophenylated thyroglobulin and thyroglobulin-beta-D-galactosidase conjugate. The immune complex formed consisting of the three components was trapped onto polystyrene balls coated with (anti-2,4-dinitrophenyl group) IgG, eluted with epsilon N-2,4-dinitrophenyl-L-lysine, and transferred onto polystyrene balls coated with (anti-human IgG gamma-chain) IgG. beta-D-Galactosidase activity bound to the last polystyrene balls was assayed by fluorometry. Anti-thyroglobulin IgG was detected in most of the patients, but not in most of the healthy subjects; levels of anti-thyroglobulin IgG in urine of the patients were well correlated to those in serum of the same patients. The measurement of anti-thyroglobulin IgG in urine by the immune complex transfer enzyme immunoassay was suggested to be useful as a diagnostic aid for autoimmune thyroid diseases. The conventional standard ELISA was not sufficiently sensitive for measuring anti-thyroglobulin IgG in urine.

Adolescent↗

Improved measurement of anti-thyroglobulin IgG in urine of patients with autoimmune thyroid diseases by sensitive enzyme immunoassay (immune complex transfer enzyme immunoassay.

Previously, antithyroglobulin IgG was assayed in dialyzed urine from patients with autoimmune thyroid diseases by a sensitive enzyme immunoassay (immune complex transfer enzyme immunoassay), and most of the assay results were useful as a diagnostic aid for autoimmune thyroid diseases. However, dialysis of urine was laborious and time-consuming, and some results were less reliable due to low levels of anti-thyroglobulin IgG in urine. This paper describes some improvements of the assay. Useful assay results could be obtained for most of urine samples without dialysis, although some interfering substance(s) was suggested to be present in some urine samples before dialysis. Accurate assay results with no interference could be obtained after gel filtration by only two min centrifugation in place of dialysis. More reliable assay results for urine samples containing low levels of antithyroglobulin IgG were obtained after concentration using a molecular sieve.

Animals↗

Principle and applications of ultrasensitive enzyme immunoassay (immune complex transfer enzyme immunoassay) for antibodies in body fluids.

The sensitivity and specificity of enzyme immunoassay for antibodies in body fluids have been improved considerably by transferring the complex of labelled antigen and antibody to be detected from one solid phase to another to eliminate interfering substance(s) in the samples (immune complex transfer enzyme immunoassay). Usefulness of the new method has been tested for antibodies in serum as well as in urine. Anti-thyroglobulin IgG could be measured not only in serum of all patients with autoimmune thyroid diseases and almost all healthy subjects but also in the urine of most of the patients. Anti-HTLV-I IgG was unequivocally demonstrated in some of sera, which were indeterminate or negative by Western blotting, and diagnosis of HIV infection by detecting anti-HIV IgG in urine and saliva would be possible with higher reliability than by conventional methods.

Antibodies↗

Immunohistochemical study of chymopapain injected into the rabbit intervertebral disc using anti-chymopapain antibody.

This study was undertaken to investigate the morphological action of chymopapain (CP) in intervertebral discs. Of 20 mature Japanese white rabbits, 19 lumbar intervertebral discs were used for electron microscopic examination, and 36 discs, for immunohistochemical examination. Discs from 3 hours to 8 weeks after injection were observed by light and electron microscopy using anti-CP rabbit antibody labeled with horseradish peroxidase. Using Kagami's DMF-dehydration, two types of fine fibers were observed in the extracellular matrix of normal nucleus pulposus, in addition to collagen fiber. After injection of CP, the thinner fibers disappeared, while the thicker fibers remained. The injected CP spread from the nucleus pulposus to the annulus fibrosus within several hours and remained as long as 4 weeks. By immunoelectron microscopy, positive granules were divided into Type 1 (lacking relationship to collagen fibers) and Type 2 (adhering to or surrounding collagen fibers). The thinner fibers appear to be proteoglycan monomer and are the target for CP.

Animals↗

[Kimura's disease (eosinophilic lymphfolliculoid granuloma)].

Eosinophilic lymphfolliculoid granuloma was first described by Kimura et al. under the heading of "On an unusual granulation accompanied by hyperplastic changes of lymphatic tissue" in 1948. There after various terms have been used for this disease, such as eosinophilic granuloma and eosinophilic lymphfolliculosis. Ever since Iizuka et al. proposed the term Kimura's disease in 1959. This disease is a relatively rare condition. It usually occurs in the soft tissue of the head and neck regions and extremities, associated with eosinophilia and elevation of serum IgE. The main histopathological features are proliferation of lymphfolliculoid structures, granulation tissue with marked infiltration of eosinophiles and mast cells, and fibrosis. The etiology of this condition is, however, still unknown. We report the clinicopathology and immunohistochemical study of this disease.

Adolescent↗

Stimulation of cholecystokinin (CCK) release from superfused rat hypothalamo-neurohypophyseal complexes by interleukin-1 (IL-1).

This in vitro study examined the effects of interleukin-1 (IL-1) and interferon-gamma (Ifn-gamma) on the release of cholecysto-kinin (CCK) from superfused hypothalamo-neurohypophyseal complexes (HNC) of rats. An increase of CCK from HNC was elicited in a dose-dependent manner by recombinant human IL-1 alpha and -1 beta in concentrations of 0.1-10 nM. In contrast, the release of CCK from HNC was not affected by recombinant human Ifn-gamma at any dose tested (0.1, 1 and 10 nM). The increased release of CCK elicited by IL-1 was calcium-dependent, as was that induced by potassium (60 mM), but it was biphasic and had a different time course and a lower magnitude than those induced by potassium and veratridine. These results suggest that IL-1 activates pituitary-adrenal axis by stimulating CCK neurons in the hypothalamus and/or neurohypophysis to release CCK, since CCK has been implicated in the regulation of adrenocorticotropin release.

Animals↗

[Ultrasensitive immunoassay of hormones].

Two methods for ultrasensitive immunoassay of peptide hormones are presented. One is to reduce the nonspecific binding of labeled reactants in two-site immunoassay by transfer of immune complexes containing labeled reactants from one solid phase to another. The other is a novel noncompetitive immunoassay method for small peptides, in which peptides are biotinylated and subsequently measured by two-site assay using anti-peptide antibody and avidin (streptavidin).

Animals↗

Sensitive enzyme immunoassay (immune complex transfer enzyme immunoassay) for (anti-human T-cell leukemia virus type I) IgG in serum using a synthetic peptide, Cys-env gp46(188-224), as antigen.

A sensitive enzyme immunoassay (immune complex transfer enzyme immunoassay) for (anti-human T-cell leukemia virus type I) IgG (anti-HTLV-I IgG) in serum using a synthetic peptide, Cys-env gp46(188-224) of HTLV-I, is described. Anti-HTLV-I IgG in test serum, which had been incubated with excess of inactive beta-D-galactosidase to eliminate interference by anti-beta-D-galactosidase antibodies, was reacted simultaneously with 2,4-dinitrophenyl-bovine serum albumin-Cys-env gp46 (188-224) conjugate and Cys-env gp46 (188-224)-beta-D-galactosidase conjugate. The complex formed consisting of the three components was trapped onto polystyrene balls coated with affinity-purified (anti-2,4-dinitrophenyl group) IgG. After washing to eliminate nonspecific IgG in the test serum and excess of the beta-D-galactosidase conjugate, the complex was eluted from the polystyrene balls with epsilon N-2,4-dinitrophenyl-L-lysine and transferred to polystyrene balls coated with affinity-purified (anti-human IgG gamma-chain) IgG. beta-D-Galactosidase activity bound to the (anti-human IgG gamma-chain) IgG-coated polystyrene balls was assayed by fluorometry. This assay was more sensitive and useful than the immune complex transfer enzyme immunoassay using Cys-Arg-env gp46(188-209) and other methods using HTLV-I as antigen.

Amino Acid Sequence↗