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Patent
Hitachi Chemical Research Center Inc. and Hitachi Ltd. | Date: 2012-12-10

Embodiments of the invention relate generally to methods of diagnosing diseases and measuring homeostatic states. In particular, the methods described here are used to characterize RNA from vesicles for expression of disease related markers. Embodiments of the invention also relate generally to the characterization of RNA by using sensitive techniques such as PCR to internally sample organ health using whole blood.


Patent
Hitachi Ltd. and Hitachi Chemical Research Center Inc | Date: 2012-03-28

The present invention provides new materials that combine the advantages of well-defined polymeric starting materials and the convenience of surface modification by physical methods into one package and, thus, offers a general and powerful platform suitable for use in numerous applications.


Patent
Hitachi Ltd., Shin Yokohama Kato Clinic and Hitachi Chemical Research Center Inc. | Date: 2012-07-18

Embodiments of the invention relate generally to ex vivo methods of quantifying expression of leukocyte-function associated mRNAs and using the quantification to characterize an individuals potential responsiveness to cancer immunotherapy. Certain embodiments relate to methods to monitor the efficacy of ongoing cancer immunotherapy by evaluating expression of leukocyte-function associated mRNAs genes before and administration of an anti-cancer immunotherapy regimen.


In order to characterize a wide spectrum of leukocyte functions with clinically applicable procedures, 0.06. ml each of heparinized whole blood was stimulated in triplicate for 4. h with phytohemagglutinin (T cell stimulator), heat aggregated IgG (IgG Fc receptor stimulator), lipopolysaccharide (toll-like receptor (TLR)-4 stimulator), zymosan (TLR-2 stimulator), monoclonal antibody against T-cell receptor alpha/beta chain, recombinant interleukin-2, and solvent controls, then 32 different leukocyte function-associated mRNAs were quantified by the method reported previously (Mitsuhashi et al. Clin. Chem. 2006). Two control genes (beta-actin, beta-2-microglobulin) were not affected by these stimulations, whereas the induction of CCL chemokines-2, 4, 8, 20, CXCL chemokines-3, 10, interleukin (IL)-8 (markers of leukocyte accumulation/recruit), granzyme B, perforin 1, tumor necrosis factor superfamily-1, 2, 5, 14, 15, CD16 (markers of cell killing), IL10, transforming growth factor beta 1 (humoral factors of immune suppression), forkhead box P3, CD25, arginase (cellular markers of immune suppression), IL2, IL4, interferon-gamma, IL17 (markers of various subsets of T helper cells), granulocyte-macrophage colony-stimulating factor (marker of antigen presenting cells), immunoglobulin heavy locus (marker of B-cells), vascular endothelial growth factor (marker of angiogenesis), pro-opiomelanocortin (marker of local pain), and CD11a mRNA (marker of leukocyte adherence to endothelium) were identified by these stimulations. The blood volume in this assay was 1.44. ml, and 4. h' incubation in whole blood was physiological. Using triplicate aliquots of whole blood for both stimulant and solvent control, statistical conclusion was drawn for each stimulant for each mRNA. The method introduced in this study will be a new paradigm for clinical cellular immunology. © 2010 Elsevier B.V. Source


Murakami T.,Hitachi Chemical Research Center Inc.
Journal of Food Protection | Year: 2012

Conventional foodborne pathogen assays currently used in the food industry often require long culture enrichments to increase pathogen levels so they can be detected. Even using sensitive real-time PCR assays, culture enrichment at least overnight is necessary especially for detection of pathogens with slow growth rates such as Listeria monocytogenes. To eliminate this cumbersome enrichment step and detect minute amounts of pathogens within 1 day, filter-based pathogen enrichment technology was developed utilizing a unique combination of glass fiber depth filter and porous filter aid materials to efficiently separate pathogens from food homogenates and avoid filter clogging by food particles. After pathogen immobilization in depth filters, only viable pathogens were selectively collected in a small volume of growth medium via microbial multiplication and migration; nonviable pathogens remained inside the filters. By assaying viable pathogens using real-time PCRs, multiple species of foodborne pathogens were detected, including L. monocytogenes, Salmonella enterica, and Escherichia coli O157:H7, at around 1 CFU/ml or 1 CFU/g in various food samples. This filter-based pathogen enrichment technology is a unique bacterial enrichment alternative to the conventional culture enrichment step and can significantly shorten the time necessary to obtain assay results. Copyright ©, International Association for Food Protection. Source

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