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Dive into the research topics where Kazuya Miyashita is active.

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Featured researches published by Kazuya Miyashita.


Molecular and Cellular Endocrinology | 1995

REGULATION OF RAT LIVER TYPE 1 IODOTHYRONINE DEIODINASE MRNA LEVELS BY TESTOSTERONE

Kazuya Miyashita; Masami Murakami; Tokuji Iriuchijima; Toshiyuki Takeuchi; Masatomo Mori

To investigate the underlying mechanisms of sex-related differences in liver type 1 iodothyronine deiodinase (ID1), we studied the sex-related differences and roles of sex steroids in liver ID1 mRNA levels in the rat. In both euthyroid and thyroidectomized rats, liver ID1 activity and ID1 mRNA levels in female rats were less than those in male rats. A positive correlation was observed between liver ID1 activity and ID1 mRNA levels. Liver ID1 activity and ID1 mRNA levels in male rats decreased after orchiectomy, and were increased to control levels by testosterone administration. Ovariectomy of beta-estradiol administration did not alter liver ID1 activity or ID1 mRNA levels in female rats. ID1 mRNA levels in cultured rat hepatocytes were significantly increased by testosterone, but not by beta-estradiol. These results suggest that the sex-related differences in liver ID1 activity are attributable to differences in ID1 mRNA levels, and that testosterone plays an important role in the sex-related differences in liver ID1 mRNA levels.


The New England Journal of Medicine | 2017

Autoantibodies against GPIHBP1 as a Cause of Hypertriglyceridemia

Anne P. Beigneux; Kazuya Miyashita; Dirk Blom; Masumi Ai; MacRae F. Linton; Weerapan Khovidhunkit; Robert Dufour; Abhimanyu Garg; Maureen McMahon; Clive R. Pullinger; Norma P. Sandoval; Xuchen Hu; Christopher M. Allan; Mikael Larsson; Tetsuo Machida; Masami Murakami; Karen Reue; Peter Tontonoz; Ira J. Goldberg; Philippe Moulin; Sybil Charriere; Loren G. Fong; Katsuyuki Nakajima; Stephen G. Young

BACKGROUND A protein that is expressed on capillary endothelial cells, called GPIHBP1 (glycosylphosphatidylinositol‐anchored high‐density lipoprotein binding protein 1), binds lipoprotein lipase and shuttles it to its site of action in the capillary lumen. A deficiency in GPIHBP1 prevents lipoprotein lipase from reaching the capillary lumen. Patients with GPIHBP1 deficiency have low plasma levels of lipoprotein lipase, impaired intravascular hydrolysis of triglycerides, and severe hypertriglyceridemia (chylomicronemia). During the characterization of a monoclonal antibody–based immunoassay for GPIHBP1, we encountered two plasma samples (both from patients with chylomicronemia) that contained an interfering substance that made it impossible to measure GPIHBP1. That finding raised the possibility that those samples might contain GPIHBP1 autoantibodies. METHODS Using a combination of immunoassays, Western blot analyses, and immunocytochemical studies, we tested the two plasma samples (as well as samples from other patients with chylomicronemia) for the presence of GPIHBP1 autoantibodies. We also tested the ability of GPIHBP1 autoantibodies to block the binding of lipoprotein lipase to GPIHBP1. RESULTS We identified GPIHBP1 autoantibodies in six patients with chylomicronemia and found that these autoantibodies blocked the binding of lipoprotein lipase to GPIHBP1. As in patients with GPIHBP1 deficiency, those with GPIHBP1 autoantibodies had low plasma levels of lipoprotein lipase. Three of the six patients had systemic lupus erythematosus. One of these patients who had GPIHBP1 autoantibodies delivered a baby with plasma containing maternal GPIHBP1 autoantibodies; the infant had severe but transient chylomicronemia. Two of the patients with chylomicronemia and GPIHBP1 autoantibodies had a response to treatment with immunosuppressive agents. CONCLUSIONS In six patients with chylomicronemia, GPIHBP1 autoantibodies blocked the ability of GPIHBP1 to bind and transport lipoprotein lipase, thereby interfering with lipoprotein lipase–mediated processing of triglyceride‐rich lipoproteins and causing severe hypertriglyceridemia. (Funded by the National Heart, Lung, and Blood Institute and the Leducq Foundation.)


Journal of Atherosclerosis and Thrombosis | 2015

Hepatic Lipase: a Comprehensive View of its Role on Plasma Lipid and Lipoprotein Metabolism

Junji Kobayashi; Kazuya Miyashita; Katsuyuki Nakajima; Hiroshi Mabuchi

Hepatic lipase (HL) is a key enzyme catalyzing the hydrolysis of triglycerides (TG) and phospholipids (PLs) in several lipoproteins. It is generally recognized that HL is involved in the remodeling of remnant, low-density lipoprotein (LDL), high-density lipoprotein (HDL) and the production of small, dense low-density lipoproteins (sd-LDLs).On the other hand, it is unclear whether HL accelerates or retards atherosclerosis. From the clinical point of view, HL deficiency may provide useful information on answering this question, but the rarity of this disease makes it impossible to conduct epidemiological study.In this review, we describe a comprehensive and updated view of the clinical significance of HL on lipid and lipoprotein metabolism.


Clinica Chimica Acta | 2015

Determination of serum lipoprotein lipase using a latex particle-enhanced turbidimetric immunoassay with an automated analyzer.

Tetsuo Machida; Kazuya Miyashita; Takuya Sone; Sayori Tanaka; Katsuyuki Nakajima; Masayuki Saito; Kimber L. Stanhope; Peter J. Havel; Hiroyuki Sumino; Masami Murakami

BACKGROUND Lipoprotein lipase (LPL) plays a central role in triglyceride-rich lipoprotein metabolism by catalyzing the hydrolysis of triglycerides. Quantification of serum LPL is useful for diagnosing lipid disorders, but there is no rapid method of measuring LPL for clinical use. METHODS We developed a rapid and sensitive latex particle-enhanced turbidimetric immunoassay (LTIA) serum LPL using latex bead-immobilized anti-LPL monoclonal antibodies. The assay was performed on a Hitachi 7700 P analyzer and evaluated for its validity as a method of quantitating the serum LPL concentration in parallel with ELISA. RESULTS Dilution tests using LTIA produced a calibration curve from 0.5 to 800ng/ml. Within-run CV was obtained in the range of 2.2-5.5%. No interference was observed in the testing of specimens containing potentially interfering substances such as bilirubin-F and C, hemoglobin, triglycerides and rheumatoid factor. A strong correlation between LTIA and ELISA was confirmed (n=40, r=0.967, y=0.99x-1.86). The normal range of LPL in pre-heparin serum was 50-77ng/ml and in post-heparin plasma 354-410ng/ml, respectively. CONCLUSION The LTIA assay is applicable in quantitating the concentration of LPL in both pre-heparin serum and post-heparin plasma. This assay is more convenient and faster than ELISA and highly suitable for clinical routine analysis.


Clinical Chemistry | 2012

ELISA System for Human Endothelial Lipase

Tatsuro Ishida; Kazuya Miyashita; Mamoru Shimizu; Noriaki Kinoshita; Kenta Mori; Li Sun; Tomoyuki Yasuda; Shigeyuki Imamura; Katsuyuki Nakajima; Kimber L. Stanhope; Peter J. Havel; Ken-ichi Hirata

BACKGROUND Endothelial lipase (EL) regulates the metabolism of HDL cholesterol (HDL-C). However, the role of EL in regulating plasma HDL-C concentrations and ELs potential involvement in atherosclerosis in humans has not been fully investigated due to the lack of reliable assays for EL mass. We developed an ELISA system for serum EL mass. METHODS Human recombinant EL proteins, purified from cultured media of human EL-transfected Chinese hamster ovary cells, were used as antigen and calibrator. Two specific monoclonal antibodies were generated in mice against recombinant EL protein for a sandwich ELISA. We measured EL mass in human serum using EL recombinant protein as a calibration standard. RESULTS The EL antibodies did not cross-react with lipoprotein lipase and hepatic triglyceride lipase. The detection limit of the ELISA was 20 pg/mL, which is approximately 10 times lower than that of previous ELISA systems. Recovery of spiked EL in serum was 90%-105%. Assay linearity was intact with a >4-fold dilution of serum. Intra- and interassay CVs were <5%. The serum EL mass in 645 human subjects was [mean (SE)] 344.4 (7.7) pg/mL (range 55.2-1387.7 pg/mL). Interestingly, serum EL mass was increased in patients with diagnosed cardiovascular disease and inversely correlated with serum HDL-C concentrations. There was no difference in EL mass between pre- and postheparin plasma samples. CONCLUSIONS This ELISA should be useful for clarifying the impact of EL on HDL metabolism and ELs potential role in atherosclerosis.


Journal of Lipid Research | 2017

Monoclonal antibodies that bind to the Ly6 domain of GPIHBP1 abolish the binding of LPL

Xuchen Hu; Mark W. Sleeman; Kazuya Miyashita; MacRae F. Linton; Christopher M. Allan; Cuiwen He; Mikael Larsson; Yiping Tu; Norma P. Sandoval; Rachel S. Jung; Alaleh Mapar; Tetsuo Machida; Masami Murakami; Katsuyuki Nakajima; Loren G. Fong; Stephen G. Young; Anne P. Beigneux

GPIHBP1, an endothelial cell protein, binds LPL in the interstitial spaces and shuttles it to its site of action inside blood vessels. For years, studies of human GPIHBP1 have been hampered by an absence of useful antibodies. We reasoned that monoclonal antibodies (mAbs) against human GPIHBP1 would be useful for 1) defining the functional relevance of GPIHBP1’s Ly6 and acidic domains to the binding of LPL; 2) ascertaining whether human GPIHBP1 is expressed exclusively in capillary endothelial cells; and 3) testing whether GPIHBP1 is detectable in human plasma. Here, we report the development of a panel of human GPIHBP1-specific mAbs. Two mAbs against GPIHBP1’s Ly6 domain, RE3 and RG3, abolished LPL binding, whereas an antibody against the acidic domain, RF4, did not. Also, mAbs RE3 and RG3 bound with reduced affinity to a mutant GPIHBP1 containing an Ly6 domain mutation (W109S) that abolishes LPL binding. Immunohistochemistry studies with the GPIHBP1 mAbs revealed that human GPIHBP1 is expressed only in capillary endothelial cells. Finally, we created an ELISA that detects GPIHBP1 in human plasma. That ELISA should make it possible for clinical lipidologists to determine whether plasma GPIHBP1 levels are a useful biomarker of metabolic or vascular disease


Clinica Chimica Acta | 2013

A new enzyme-linked immunosorbent assay system for human hepatic triglyceride lipase.

Kazuya Miyashita; Junji Kobayashi; Shigeyuki Imamura; Noriaki Kinoshita; Kimber L. Stanhope; Peter J. Havel; Katsuyuki Nakajima; Tetsuo Machida; Hiroyuki Sumino; Makoto Nara; Masami Murakami

BACKGROUND The objective of this study was to establish a new sandwich based enzyme linked immunosorbent assay (ELISA) for measuring the protein mass of human hepatic triacylglyceride lipase (HTGL). METHOD Two mouse monoclonal antibodies raised against human HTGL were used for the sandwich ELISA. The post-heparin plasma (PHP) samples obtained at a heparin dose of 50 unit/kg from 124 normolipidemic subjects were used for this ELISA. RESULTS The dynamic assay range of the developed ELISA for the HTGL was from 0.47 to 30 ng/ml. The CV was <7% in both intra- and inter-assays, and it did not cross-react with lipoprotein lipase or endothelial lipase (EL). The HTGL concentration in PHP showed a strong correlation with HTGL activity [n=121, r=0.778, p<0.001]. There was a weak relation of HTGL concentration against high-density lipoprotein cholesterol (HDL-C) [n=123, r=-0.229, p=0.011] but no relations against total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), triglycerides (TG), small dense LDL, remnant like particles cholesterol (RLP-C) and RLP-TG were confirmed. Interestingly, a weak but positive correlation between HTGL concentration and EL concentration was shown [n=122, p=0.013, r=0.224]. CONCLUSION These results indicate that this new sandwich ELISA for measuring HTGL concentration in PHP can be applied in a daily clinical practice.


Clinica Chimica Acta | 2016

The majority of lipoprotein lipase in plasma is bound to remnant lipoproteins: A new definition of remnant lipoproteins.

Koichi Sato; Fumikazu Okajima; Kazuya Miyashita; Shigeyuki Imamura; Junji Kobayashi; Kimber L. Stanhope; Peter J. Havel; Tetsuo Machida; Hiroyuki Sumino; Masami Murakami; Ernst J. Schaefer; Katsuyuki Nakajima

BACKGROUND Lipoprotein lipase (LPL) is a multifunctional protein and a key enzyme involved in the regulation of lipoprotein metabolism. We determined the lipoproteins to which LPL is bound in the pre-heparin and post-heparin plasma. METHODS Tetrahydrolipstatin (THL), a potent inhibitor of serine lipases, was used to block the lipolytic activity of LPL, thereby preventing changes in the plasma lipoproteins due to ex vivo lipolysis. Gel filtration was performed to obtain the LPL elution profiles in plasma and the isolated remnant lipoproteins (RLP). RESULTS When ex vivo lipolytic activity was inhibited by THL in the post-heparin plasma, majority of the LPL was found in the VLDL elution range, specifically in the RLP as inactive dimers. However, in the absence of THL, most of the LPL was found in the HDL elution range as active dimers. Furthermore, majority of the LPL in the pre-heparin plasma was found in the RLP as inactive form, with broadly diffused lipoprotein profiles in the presence and absence of THL. CONCLUSIONS It is suggested that during lipolysis in vivo, the endothelial bound LPL dimers generates RLP, forming circulating RLP-LPL complexes in an inactive form that subsequently binds and initiates receptor-mediated catabolism.


Neuropeptides | 1990

Stimulation by a TRH precursor, TRH-Gly, of TSH and PRL secretion in rats: Effect of starvation

M. Mori; Masami Murakami; Tokuji Iriuchijima; Kazuya Miyashita; Teturou Satoh; T. Monden; Toshio Michimata; Isao Kobayashi; S. Kobayashi

The hypophysial activities of a possible direct precursor of thyrotropin (TSH)-releasing hormone (TRH), TRH-Gly, were evaluated in estrogen, progesterone-primed rats under urethane anesthesia. Intravenous administration of TRH-Gly in doses of 2-200 micrograms caused a significant and dose-dependent increase in blood TSH and prolactin (PRL). The stimulatory activity of TRH-Gly was 170 to 400-times less potent than that of TRH. The lower potency was confirmed by the action of TRH-Gly on the anterior pituitary cells in vitro. In starved rats, TRH-Gly apparently stimulated TSH and PRL secretion in a dose-dependent manner, and the stimulatory activity increased in starved rats as compared to normal controls. TRH-Gly did not affect [3H-MeHis]TRH binding in pituitary plasma membranes. These data imply that large amounts of TRH-Gly may have significant biological activities and these are potentiated in the starved condition.


Journal of Lipid Research | 2016

An LPL-specific monoclonal antibody, 88B8, that abolishes the binding of LPL to GPIHBP1.

Christopher M. Allan; Mikael Larsson; Xuchen Hu; Cuiwen He; Rachel S. Jung; Alaleh Mapar; Constance Voss; Kazuya Miyashita; Tetsuo Machida; Masami Murakami; Katsuyuki Nakajima; André Bensadoun; Loren G. Fong; Stephen G. Young; Anne P. Beigneux

LPL contains two principal domains: an amino-terminal catalytic domain (residues 1–297) and a carboxyl-terminal domain (residues 298–448) that is important for binding lipids and binding glycosylphosphatidylinositol-anchored high density lipoprotein binding protein 1 (GPIHBP1) (an endothelial cell protein that shuttles LPL to the capillary lumen). The LPL sequences required for GPIHBP1 binding have not been examined in detail, but one study suggested that sequences near LPL’s carboxyl terminus (residues ∼403–438) were crucial. Here, we tested the ability of LPL-specific monoclonal antibodies (mAbs) to block the binding of LPL to GPIHBP1. One antibody, 88B8, abolished LPL binding to GPIHBP1. Consistent with those results, antibody 88B8 could not bind to GPIHBP1-bound LPL on cultured cells. Antibody 88B8 bound poorly to LPL proteins with amino acid substitutions that interfered with GPIHBP1 binding (e.g., C418Y, E421K). However, the sequences near LPL’s carboxyl terminus (residues ∼403–438) were not sufficient for 88B8 binding; upstream sequences (residues 298–400) were also required. Additional studies showed that these same sequences are required for LPL binding to GPIHBP1. In conclusion, we identified an LPL mAb that binds to LPL’s GPIHBP1-binding domain. The binding of both antibody 88B8 and GPIHBP1 to LPL depends on large segments of LPL’s carboxyl-terminal domain.

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Junji Kobayashi

Kanazawa Medical University

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