Hitachi Chemical Research Center

Irvine, CA, United States

Hitachi Chemical Research Center

Irvine, CA, United States
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Mitsuhashi M.,Hitachi Chemical Research Center | Taub D.D.,U.S. National Institute on Aging | Kapogiannis D.,U.S. National Institute on Aging | Eitan E.,U.S. National Institute on Aging | And 6 more authors.
FASEB Journal | Year: 2013

Amyloid-1-42 (A) peptide effects on human models of central nervous system (CNS)-patrolling macrophages (Ms) and CD4 memory T-cells (CD4- Tms) were investigated to examine immune responses to A in Alzheimer's disease. A and lipopolysaccharide (LPS) elicited similar M cytokine and exosomal mRNA (ex-mRNA) responses. A- and LPS-stimulated Ms from 20 >65-yr-old subjects generated significantly more IL-1, TNF, and IL-6, but not IL-8 or IL-12, and significantly more ex-mRNAs for IL-6, TNF, and IL-12, but not for IL-8 or IL-1, than Ms from 20 matched 21- to 45-yr-old subjects. CD4-Tm generation of IL-2, IL-4, and IFN and, for young subjects, IL-10, but not IL-6, evoked by A was significantly lower than with anti-T-cell antigen receptor antibodies (Abs). Abs significantly increased all CD4-Tm ex-mRNAs, but only IL-2 and IL-6 ex-mRNAs were increased by A. There were no significant differences between cytokine and ex-mRNA responses of CD4-Tms from the old compared to the young subjects. M-derived serum exosomes from the old subjects had significantly higher IL-6 and IL-12 ex-mRNA levels than those from the young subjects, whereas there were no differences for CD4-Tm-derived serum exosomes. An A level relevant to neurodegeneration elicited broad M cytokine and exmRNA responses that were significantly greater in the old subjects, but only narrow and age-independent CD4-Tm responses.-Mitsuhashi, M., Taub, D. D., Kapogiannis, D., Eitan, E., Zukley, L., Mattson, M. P., Ferrucci, L., Schwartz, J. B., Goetzl, E. J. Aging enhances release of exosomal cytokine mRNAs by A1-42-stimulated macrophages. © FASEB.

Omori K.,Beckman Research Institute | Mitsuhashi M.,Hitachi Chemical Research Center | Ishiyama K.,Beckman Research Institute | Nair I.,Beckman Research Institute | And 4 more authors.
Diabetologia | Year: 2011

Aims/hypothesis: TNF-α plays important roles in the pathogenesis of type 1 and type 2 diabetes mellitus. In light of this, we examined the involvement of a pro-apoptotic gene, BBC3 (also known as PUMA), in TNF-α-mediated beta cell dysfunction and destruction in human islets. Methods: Human islets were exposed in vitro to TNF-α alone or in combination with IFN-γ. Gene expression was assessed by RT-PCR using a set of single islets. Protein abundance and cellular localisation of BBC3 were assessed by immunoblot and immunohistochemistry. A marginal number of islets were transplanted into diabetic NODscid mice to correlate in vivo islet function with BBC3 expression. Results: BBC3 and IL8 mRNA were upregulated in TNF-α-stimulated islets in a dose-dependent manner and enhanced through addition of IFN-γ, but not upregulated by IFN-γ alone. Immunohistochemistry revealed that TNF-α in combination with IFN-γ upregulated basal BBC3 abundance in the cytoplasm of beta cells along with the perinuclear clustering of mitochondria partially co-localised with BBC3. TNF-α alone did not induce beta cell death, but did abrogate preproinsulin precursor mRNA synthesis in response to high glucose stimulation, which was inversely associated with upregulation of BBC3 mRNA expression by TNF-α. Higher BBC3 mRNA expression in islets correlated with decreased graft function in vivo. Conclusions/interpretation: These results suggest that BBC3 mRNA can serve as a molecular marker to detect early TNF-α-induced beta cell stress and may help identify islet-protective compounds for the treatment of diabetes. © 2011 Springer-Verlag.

Hitachi Ltd., Hitachi Chemical Research Center and The Regents Of The University Of California | Date: 2013-10-02

Embodiments of the invention relate generally to methods of identifying subjects likely to develop diabetes-associated damage to the nephron, or subjects in the early stages of diabetic nephropathy. In particular, several embodiments relate to quantification of diabetic nephropathy-associated RNA isolated from vesicles from patient urine samples is performed to compare a subject to a population having normal nephron function and/or to track progression of diabetic nephropathy in said subject over time.

Twardowski P.W.,City of Hope National Medical Center | Beumer J.H.,University of Pittsburgh | Chen C.S.,Loma Linda University | Kraft A.S.,Medical University of South Carolina | And 5 more authors.
Anti-Cancer Drugs | Year: 2013

There is a need for efficacious therapies for metastatic castration-resistant prostate cancer (mCRPC) after disease progression on docetaxel. The SRC tyrosine kinase and its related family members may be important drivers of prostate cancer and can be inhibited by dasatinib. mCRPC patients, after one previous chemotherapy, started dasatinib at 70 mg twice daily, amended to 100 mg daily. The primary endpoint was the disease control (DC) rate, defined as complete response (CR), partial response (PR), or stable disease (SD) in prostate specific antigen (PSA), RECIST, bone scan, and FACT-P score. Up to 41 patients were to be accrued (two-stage design, 21+20) to rule out a null-hypothesized effect of 5 versus 20% (α=0.05, β=0.1). Secondary endpoints included progression-free survival, toxicity, and pharmacokinetic and pharmacodynamic correlatives. Of 38 patients, 27 were evaluable for response or toxicity. The median duration of therapy was 55 days (6-284). Five patients showed DC after 8 weeks of therapy (18.5% DC, 95% CI: 6.3-38.1%). One PR (3.7% response rate, 95% CI: 0.1-19.0%) was observed in a patient treated for 284 days. Twelve patients (43%) discontinued treatment for toxicity. Dasatinib induced a decrease in phytohemagglutinin-stimulated CSF2, CD40L, GZMB, and IL-2 mRNAs in blood cells, indicating target engagement. Decreases in plasma IL-6 and bone alkaline phosphatase, and in urinary N-telopeptide, were associated with DC. Dasatinib has definite but limited activity in advanced mCRPC, and was poorly tolerated. The observation of a patient with prolonged, objective, clinically significant benefit warrants molecular profiling to select the appropriate patient population. © 2013 Wolters Kluwer Health | Lippincott Williams & Wilkins.

Rosa J.S.,University of California at Irvine | Rosa J.S.,Clinical Translational Science Institute | Mitsuhashi M.,Hitachi Chemical Research Center | Oliver S.R.,University of California at Irvine | And 6 more authors.
Diabetes/Metabolism Research and Reviews | Year: 2010

Background: Abnormal systemic concentrations of proinflammatory cytokines/chemokines have been implicated in the development of longterm cardiovascular complications in type 1 diabetes (T1DM) and obesity. Whether leukocyte white blood cell (WBC) gene expression of these proinflammatory mediators contributes to their increased systemic levels, however, remains unclear, especially in the pediatric patient populations. This study examines mRNA changes of 9 cytokines and chemokines in WBCs following ex vivo immunostimulation from 9 T1DM (13.4 ± 0.5 year, 4F/5 M), 23 overweight (OW, 12.3 ± 0.5 year, 10F/13M, BMI% 97.1 ± 0.5 and >90.0), and 21 healthy (CL, 13.8 ± 0.7 year, 9F/12 M, BMI% 59.6 ± 4.6 and <85.0) children. Methods: All subjects had been maintained in euglycemic conditions for at least 90 min before blood draws. Whole blood was then sampled and incubated with anti-T-cell receptor (TCR) antibody or heat-aggregated IgG (HAG) to stimulate T-cell and Fc receptors (FcR), respectively. After lysis of leukocytes, mRNA levels of six tumor necrosis factor superfamily cytokines (TNFSF2, 5, 6, 7, 9, 14) and three chemokines (CCL8, 20, and CXCL10) were measured using RT-PCR. Results: Following TCR stimulation, T1DM displayed significantly greater mRNA responses than CL for TNFSF5, 7, 9, and CCL8, and CXCL10; TNFSF9, CCL8, and CXCL10 were also significantly higher in T1DM than OW; no difference was observed between OW and CL. FcR stimulation induced similar responses across groups. Conclusions: Leukocytes of T1DM children displayed exaggerated gene expression in response to ex vivo TCR induction of five key proinflammatory cytokines/chemokines. This elevated leukocyte gene expression may be one of the pathophysiological contributors to the development of vascular complications in T1DM. Copyright © 2009 John Wiley & Sons, Ltd.

Aoki J.,Tokyo Metropolitan Cancer and Infectious Diseases Center Komagome Hospital | Ohashi K.,Tokyo Metropolitan Cancer and Infectious Diseases Center Komagome Hospital | Mitsuhashi M.,Hitachi Chemical Research Center | Murakami T.,Hitachi Chemical Research Center | And 5 more authors.
Clinical Chemistry | Year: 2014

BACKGROUND: Bone marrow (BM) aspiration often can be a painful medical procedure. It is unavoidable, however,because hematopoietic precursor cells (HPC) exist only inBMand few escape to peripheral blood (PB). We hypothesized that HPCs might release exosomes and microvesicles (EMV) in BM, and the resulting EMV would penetrate into PB. Such BM-derived EMV might be identified in PB by measuring specific mRNAs produced by HPC. METHODS: Human plasma was applied to an EMVcapture filter plate. After centrifugation, captured EMV were lysed on the filter plate. Resulting lysates were transferred to an oligo(dT)-immobilized microplate for mRNA isolation followed by reverse transcription PCR (RT-PCR). Using this system, myeloid-, erythroid-,and megakaryocyte-lineage-specific poly(A)+mRNAswere quantified in plasma obtained from 18 patients whohad undergone hematopoietic stem cell transplantation (HSCT). RESULTS: When fluorescent liposomes were applied to the filter plate, more than 95% of applied liposomes were absorbed. When human plasma was applied, a scanning electron microscope showed EMV-like particles on the membrane of the filter plate. After RT-PCR,various HPC-specific mRNAs were detected, and the results were equivalent to those derived from the standardultracentrifugation method. The levels of thesemRNAs were undetectable after HSCT and became detectable 1-2 weeks after HSCT, a substantially earlier time point than with traditional hematological analysis. The recovery of EMV mRNA at day 15 corresponded to the final clinical outcome at day 180. CONCLUSIONS: HPC-derived mRNAs in plasma EMV may represent new biomarkers for the assessment of BM condition and could reduce the necessity for frequent BM aspiration. © 2013 American Association for Clinical Chemistry.

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