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Biomedical subjects

Y Ichikawa

Publications and source records attributed to Y Ichikawa.

At least 739 records · Page 41Linked to original sources

Sensitive and simplified method for the differential determination of serum levels of ketone bodies.

A highly sensitive and simplified method for the differential determination of serum ketone bodies has been developed. Serum was deproteinized with perchloric acid, and acetoacetate contained in the supernate was reacted with newly synthesized p-nitrobenzene diazonium fluoroborate at 37 degrees C for 10 min. The formed hydrazo compound was converted by alkali to the more stable azo compound which has a peak absorbance at 645 nm. For the determination of 3-hydroxybutyrate, this was enzymatically converted to acetoacetate using 3-hydroxybutyrate dehydrogenase, LDH, NAD and pyruvate. Using 0.2 ml serum, acetoacetate and 3-hydroxybutyrate could be quantitated in 30 min. The described method is five times more sensitive than the enzymatic photometric method and can detect individual ketone bodies at concentrations as low as 20 mumol/l. Differential determination of serum levels of ketone bodies is clinically useful for the diagnosis of type 1 diabetes and in monitoring diabetic control.

3-Hydroxybutyric Acid↗

Phosphorylation of the myosin heavy chain. Its effect on actin-activated Mg2+-stimulated ATPase in leukaemic myeloblasts.

Myosin purified from a murine myeloid leukaemia cell line (M1) that had been incubated with [32P]orthophosphate incorporated 32P into the heavy, but not the light, chain. When the heavy chain was dephosphorylated by bacterial alkaline phosphatase, myosin that had low actin-activated ATPase activity gained higher activity only in the presence of the light-chain kinase. In the absence of the light-chain kinase, however, the Mg2+-stimulated ATPase activity of myosin was not activated by actin, regardless of phosphatase treatment. These results indicate that the activity of M1 myosin ATPase is regulated by phosphorylation of both the light and heavy chains. A scheme for this regulation by phosphorylation is presented and discussed.

Actins↗

Reactivity of mitogen-induced autorosette-forming cells to interleukin 2 (T-cell growth factor).

The reactivity of mitogen-induced autologous rosette-forming cells (ARFC) to interleukin 2 (IL-2; T-cell growth factor) was studied in the present report. Both ARFC-enriched T cells and ARFC-depleted T cells, which were separated from concanavalin A (Con A)-activated T cells, were reactive to this factor. The IL-2 activity was absorbed by both ARFC-enriched and ARFC-depleted T cells, although ARFC-enriched T cells could absorb more IL-2 activity. Furthermore, ARFC were further inducible by IL-2 from non-ARFC. These results suggest the expression of the receptors for IL-2 on both ARFC and non-ARFC following mitogen stimulation. They further support the possibility that mitogen-induced ARFC, rather than being recruited only from such a minor T-cell subset as the spontaneous ARFC, are more likely the result of most T cells being responsive to mitogenic stimulation and expressing the receptors for autologous erythrocytes by the effects of IL-2 and mitogen.

Adult↗

Endogenous peroxidase activity in brush cell-like cells in the large intestine of the bullfrog tadpole, Rana catesbeiana.

A special cell type was identified in the mucosal epithelium of the large intestine of the tadpole of the bullfrog, Rana catesbeiana. It is a slender, columnar cell, with a dark, basally situated nucleus. By electron microscopy the cell displays prominent bundles of filaments emerging from each microvillus and extending deep into the cytoplasm without ending in the terminal web. It has longer and more crowded microvilli than the absorptive cell. The specialized cell is also characterized by the presence of many apical vesicles and numerous subapical dense bodies. These cytological features suggest that it may be a brush cell (Rhodin and Dalhamn 1956). These cells displayed endogenous peroxidase activity in smooth and rough endoplasmic reticulum, in the well-developed Golgi apparatus and in apical vesicles. Furthermore, peroxidase reaction product was frequently observed on their luminal surface membrane. These findings suggest that the brush cell in the large intestine of the bullfrog tadpole may be a secretory cell.

Animals↗

Pharmacokinetics of bredinin in renal transplant patients.

A pharmacokinetic study of bredinin, a new immunosuppressive agent, was carried out in 28 renal transplant patients. Serum bredinin concentration-time curves were analyzed using a one-compartment open model with a first order absorption process. The peak serum bredinin level appeared 2.4 h after oral administration of bredinin 50-200 mg. The calculated mean peak serum level was 0.852 micrograms/ml/mg/kg, when the dose was adjusted to the body weight of the patient. In the dosage range used of 0.85-4.46 mg/kg, a linear relationship was observed between the dose and the peak serum bredinin level. The elimination rate of bredinin from serum was dependent on kidney function, and the elimination rate constant was well correlated with the endogenous creatinine clearance. No circadian rhythm was apparent in the elimination rate constant. The absorption rate of bredinin from the gastrointestinal (GI) tract was affected by GI diseases. The need for dosage adjustment based on the renal function of the transplant patient is suggested.

Adolescent↗

Further characterization of mitogen-induced autorosette-forming cells: correlation with T-cell subsets and with lymphocyte proliferative responses to mitogens.

Spontaneous autologous rosette-forming cells (ARFC), which form rosettes with autologous erythrocytes, have been of interest as a subset of thymus-derived lymphocytes (T cells). An association of these cells with concanavalin A (Con A)-induced ARFC has been suggested. Furthermore, the Con A-induced ARFC have been shown to be a suppressor T-cell subset in the Con A-generated suppressor system. We have previously reported the induction of ARFC from T cells by several T-cell mitogens such as phytohemagglutinin-P (PHA) and allogeneic non-T cells other than Con A. In the present report, we further characterized the mitogen-induced ARFC and have extended the study to patients with systemic lupus erythematosus (SLE). We have found that ARFC are also inducible from peripheral blood T cells by pokeweed mitogen (PWM). Studies of T-cell surface markers on the ARFC using OKT monoclonal antibodies confirmed the induction of ARFC from both OKT4- and OKT8-reactive T cells by either Con A, PHA, or PWM stimulation. However, OKT4-reactive T cells were the major cellular source of the ARFC induced by all of the mitogens. In studies of SLE patients, proportions of both Con A- and PWM-induced ARFC were found to be significantly low in PBL of SLE patients treated with moderate or large doses of prednisone, with or without concomitant immunosuppressants, but not in SLE patients without such treatment. Proportional analysis of the T cells and their subsets suggested association of these alterations in the mitogen-induced ARFC with the OKT4-reactive T cells, since a significant decrease in the OKT4-reactive T-cell subset was demonstrated in the PBL of these patients. Proportions of PHA-induced ARFC, however, were not significantly different between SLE patients and healthy adults. Moreover, positive correlations of the mitogen-induced ARFC with lymphocyte proliferative responses to each mitogen were established in both SLE patients and healthy adults. These results further support our previous observation that suggest the receptors for autologous erythrocytes are enhanced or reexpressed on those T cells which are highly activated by mitogens.

Antibodies, Monoclonal↗

Importance of increased urinary calcium excretion in the development of secondary hyperparathyroidism of patients under glucocorticoid therapy.

Parathyroid function and calcium metabolism were studied in 44 patients under glucocorticoid therapy (steroid group) and in 25 control subjects. Nephrogenous cAMP and serum immunoreactive parathyroid hormone levels in the steroid group were significantly higher than those in control subjects (p less than 0.001). Nephrogenous cAMP in the steroid group correlated positively with prednisolone dosage (r = 0.424, p less than 0.01), and most patients who showed obvious elevations of nephrogenous cAMP had received over 10 mg/day of prednisolone for at least 2 mo. Fasting urinary calcium in the steroid group [166.1 +/- 78.5 (+/- SD) mg/g creatinine] was about 2 times greater than that in control subjects (74.1 +/- 35.6) (p less than 0.001). Fasting urinary calcium in control subjects correlated negatively with nephrogenous cAMP (r = -0.486, p less than 0.02). In contrast, these values in steroid group showed significant positive correlation (r = 0.631, p less than 0.001), suggesting that increased urinary calcium excretion is an important factor in the development of secondary hyperparathyroidism. Elevated nephrogenous cAMP and serum immunoreactive parathyroid hormone levels decreased after the administration of trichlormethiazide and/or 1 alpha hydroxy-vitamin D3. We conclude that increased urinary calcium excretion plays an important role in the development of secondary hyperparathyroidism in patients under glucocorticoid therapy and that the administration of thiazide and/or vitamin D could improve the secondary hyperparathyroidism caused by glucocorticoid therapy.

Adolescent↗

Tissue and subcellular distributions of cholecalciferol 25-hydroxylase: cytochrome P-450D25-linked monooxygenase system.

Cholecalciferol (vitamin D3) 25-hydroxylase activity and its tissue and subcellular distributions were studied using rabbit, rat, bovine, chick, duck, guinea-pig and yellowtail. Vitamin D3 25-hydroxylase was found not only rabbit and bovine livers, kidneys and intestines, but also in lungs, pituitary glands, adrenocortices, ovaries, adipose tissues, blood vessels, brains and testes. Subcellular fractions of rabbit livers were examined for the presence of this activity and it was detected in the nuclear membranes, smooth and rough endoplasmic reticula, Golgi membranes and mitochondrial membranes. The activity in microsomes was induced by sodium phenobarbital, but not by beta-naphthoflavone and 3-methylcholanthrene, and was inhibited by isocyanide and metyrapone as inhibitor of cytochrome P-450.

Animals↗

Purification and characterization of renal ferredoxin from bovine renal mitochondria.

A renal ferredoxin was purified from bovine renal mitochondria to electrophoretic purity. The molecular weight of the renal ferredoxin was estimated by gel filtration and SDS-polyacrylamide gel electrophoresis to be 12,500 and 13,000, respectively. The optical absorption spectrum of renal ferredoxin in the oxidized form showed two peaks at 416 and 457 nm in the visible region, and the EPR absorption spectrum showed peaks at gx = gy =1.94 and gz = 2.02 in the reduced form at 13K. These spectra were typical of the 2S-2Fe type ferredoxins. Dissimilarities were recognized in the amino acid composition and isoelectric point between bovine renal ferredoxin and bovine adrenodoxin, but not in the optical, magnetic, and immunochemical properties. The reconstitution of 25-hydroxyvitamin D3-1 alpha-hydroxylase system was performed with the three components of NADPH-adrenodoxin reductase from bovine adrenal mitochondria, renal ferredoxin, and cytochrome P-450(1) alpha from bovine renal mitochondria. The results showed that the renal ferredoxin was essential for the 1 alpha-hydroxylase activity of 25-hydroxyvitamin D3.

Animals↗

Elicitation of delayed type hypersensitivity in chicks after in ovo sensitization with different molecular forms of the same hapten.

Lymphocyte sensitization, which participates in delayed type hypersensitivity (DTH) in chick embryos, was studied. The in ovo injection of dinitrophenylated keyhole limpet hemocyanin (DNP-KLH) or DNP-dextran (DNP-D) led to delayed onset of the hapten-specific reaction as shown by allergic contact dermatitis (ACD) after hatching. The extent of the ACD response was not directly dependent on the antigen dosage or the number of injections given for sensitizing. The magnitude of the ACD response was higher in chicks sensitized with DNP-D than in those given DNP-KLH. These findings suggest the presence of embryonic lymphocytes which can be sensitized by in ovo antigenic stimulation at the later stage of embryogenesis and may make possible the differentiation of functional lymphocytes. Antigen stimulation with higher doses may be inadequate for the in ovo sensitization of embryonic lymphocytes. The ACD response elicited by DNFB in chicks primed with either DNP-KLH or DNP-D was thought to be T-cell dependent, from the kinetics of the ACD peak within a period of 24 to 48 hr. Furthermore the conditions for in ovo sensitization of embryonic lymphocytes by DNP-D seem to be different from those for sensitization by DNP-KLH.

Age Factors↗

Sequential serum measurements of thyrotropin binding inhibitor immunoglobulin G in transient familial neonatal hypothyroidism.

Infants with transient neonatal hypothyroidism, in whom TSH binding inhibitor immunoglobulin G (IgG) (TBII) were sequentially measured, are described. Their mother had been taking thyroid replacement for hypothyroidism due to nongoitrous autoimmune thyroiditis. IgGs inhibiting TSH binding were detected in maternal sera by radioreceptor assay. These IgGs also inhibited the adenylate cyclase response to TSH in human thyroid membranes. Three infants had frank hypothyroidism immediately after birth, and TBII were detected in two of them. In the two surviving infants, hypothyroidism was transient and improved when TBII disappeared from their sera. The profile of TBII in one patient corresponded to the IgG disappearance curve. These findings suggest that the transient neonatal hypothyroidism reported was caused by transplacental transfer of TBII.

Adenylyl Cyclase Inhibitors↗

Changes in actin-related gelation of crude cell extracts during differentiation of myeloid leukemia cells.

Gelation of extracts of a myeloid leukemia cell line (Ml) was compared before and after differentiation induced with conditioned medium (CM) from rat embryo cells. Although an extract of Mml cells, a macrophage line derived from Ml line, gelled when warmed in the presence of 2 mM MgCl2, undifferentiated Ml cells gelled only after dialysis and a supplement of exogenous actin. After differentiation had been induced, an addition of exogenous actin, but not dialysis, was needed for gelation. Small amounts of KCl always inhibited the gelation of the control Ml cell extracts, but they promoted gelation of the CM-treated Ml and Mml cell extracts. Thus, the dialysis required for gelation of the control Ml cell extract appears to be necessary for the exclusion of endogenous KCl. Several possible mechanisms for the KCl control of gelation, as well as different requirements of exogenous actin needed for gelation are discussed based on the results of our experiments.

Actins↗

[Bronchopulmonary diseases due to ultra-hard metal dust, with special reference to the result of the dust analysis].

In order to clarify the relationship between exposure to hard metal dust containing cobalt and bronchopulmonary diseases, cross-sectional medical and occupational hygienic investigations were conducted in a hard metal tool manufacturing factory. As a part of the environmental investigation, general air concentrations and personal exposure to both total dust and cobalt dust were measured. The highest total dust and cobalt concentrations in general atmosphere and personal samples were found in a powder handling room among all departments. The highest personal exposure concentration of cobalt was 1.29 mg/m3 (arithmetic mean value) in the dry grinder using diamond wheels. Powder handlers were exposed to cobalt 0.56 mg/m3, press operators to 0.66 mg/m3, shapers to 0.19 mg/m3, wet grinders to 0.07 mg/m3, and sintering workers to 0.03 mg/m3. In Japan TLV for cobalt has not yet been established. Dust exposure measurement should be carried out in all hard metal industries. TLV for cobalt is discussed by means of literature.

Air Pollutants, Occupational↗

Mycoplanecins, novel antimycobacterial antibiotics from Actinoplanes awajinensis subsp. mycoplanecinus subsp. nov. II. Isolation, physico-chemical characterization and biological activities of mycoplanecin A.

New antibiotics, mycoplanecins, were found in the culture broth of an actinomycete identified as Actinoplanes awajinensis subsp. mycoplanecinus subsp. nov. Mycoplanecin complex was extracted with organic solvents both from the culture filtrate and mycelium and purified by column chromatography on silica gel and Florisil. Mycoplanecin A, a major component, was separated by high performance liquid chromatography on Prep PAK-500/C18 column. The physico-chemical characterization revealed that mycoplanecin A was a new cyclic peptide antibiotic. Mycoplanecins exhibited strong activities primarily against mycobacteria and related microorganisms.

Actinomycetales↗

[Changes of cellular contractile proteins associated with differentiation of leukemic cells].

A myeloid leukemia cell line, M1, can be induced to differentiate to macrophages and gains motile and phagocytic functions which are not present before differentiation. The appearance of these differentiated functions were always accompanied with a loss of mitotic activity. It is well known that the intracellular contractile proteins play an important role in both cell motility and cytokinesis. Our question is, why the actomyosin system in the M1 cell line is utilized only for cell division and not for cell motility before differentiation, and why the opposite situation takes place after differentiation. Changes in contractile proteins hitherto revealed during the differentiation of this cell line are as follows: Actin; contents, synthetic rate, ability to polymerize, ratio of gamma-actin, activity to activate the myosin Mg2+ ATPase, and binding to plasma membrane all showed an increase after differentiation. Changes in the chemical structure of actin molecule were suggested by peptide mapping. In the undifferentiated M1 cytoplasms, an inhibitor protein for actin polymerization was identified. Myosin; contents, ratio of light chain L2, and binding to plasma membrane showed an increase after differentiation. Which one plays the most essential role in the transition from cell division to cell motility, and what interrelation is there among those changes are remained to be disclosed.

Actins↗