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Flow Cytometer Performance Characterization, Standardization and Calibration against CD4 on T Lymphocytes Enables Quantification of Biomarker Expressions for Immunological Applications

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Journal of Biomedical Science and Engineering 2014年第9期7卷 756-768页

作者： Heba Degheidy steven bauer Gerald Marti Lili Wang Center for Biologics Evaluation and Research US Food and Drug Administration (FDA) Bethesda Maryland USA Center for Devices and Radiological Health FDA Silver Spring Maryland USA National Institute of Standards and Technology (NIST) Gaithersburg Maryland USA

There is an urgent need for developing a procedure for biomarker standardization and relative quantificationin clinical laboratories. Measuring the expression levels of cell antigens is critical for the diagnosis of many diseases, e.g. leukemia, lymphoma and immunodeficiency diseases. One of the most significant challenges in flow cytometry is obtaining inter-laboratory and intra-laboratory consistent and reproducible results across multiple cytometer platforms and locations longitudinally over time. To obtain measurement consistency, the target flow cytometer voltages should be optimized to segregate the negative population from the electronic noise, and to keep the brightest positive population within the dynamic range of each detector. Then target values should be determined and transferred to selected cytometers. In this study, we optimized a procedure for instrument standardization across three different flow cytometer platforms from the same vendor and in two different locations. The biomarker quantification was implemented on standardized instruments using CD4 expression on T lymphocytes with a known amount of antibody bound per cell as a quantification standard. Our results on blood cell subset typing and CD19 quantification demonstrated that consistent and reliable results could be accomplished between instruments using the developed procedure. Quantitating the expression levels of certain cell biomarkers relative to a known reference marker before, during, and after therapy would provide important information for monitoring antibody-based therapy and could be potentially used to adjust dosing. Presently, we are implementing this protocol to quantify critical disease biomarkers, and making necessary modifications to the procedure to include instruments from different instrument manufacturers.

关键词： Flow Cytometry Biomarker Quantification Instrument Standardization Target Median Fluorescence Intensity (MFI)

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A predictive accident-duration based decision-making module for rerouting in environments with V2V communication

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Journal of Traffic and Transportation Engineering(English Edition) 2017年第6期4卷 535-544页

作者： besat zardosht steven s.beauchemin michael a.bauer department of computer science western universityLondonOntario N6A 3K7Canada

Communication for cooperative traffic management increases convenience and efficiency in driving. Intelligent vehicles can collect information about the driving environment, the driver situation and, more importantly, information on other vehicles. While giving this information to the driver can be useful, there is also the possibility of presenting the driver with too much information. Existing vehicles already have some mechanisms to take certain actions if the driver fails to act. Future vehicles will need more complex decision-making modules which receive the raw data from all available sources, process this data and inform the driver about existing or impending situations and suggest, or even take actions. In this work, we explore the use of a decision-making module for accident situations that processes information from VANET communication and advises the driver based on the situation. Our decision-making algorithm is a simple and effective algorithm that can be implemented in each vehicle to assist the driver in situations where rerouting to avoid traffic congestion caused by an accident may be necessary. Our approach estimates accident duration and provides a rerouting for upcoming traffic. Our algorithm has been implemented and evalu- ated within an elegant traffic simulation system where both city and highway environments are considered, along with various proportions of V2V-equipped vehicles. Overall, the results indicate that using our decision-making module in vehicles shows great potential for improving performance of vehicular systems by reducing both travel and wait times and providing more accurate information on surrounding environments.

关键词： Cooperative traffic managementsystemVANETDecision-making systemAccident duration model

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Male Tibetan macaques' (Macaca thibetana) choice of infant bridging partners

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Zoological Research 2014年第3期35卷 222-230页

作者： Briana bauer Lori K. SHEERAN Megan D. MATHESON Jin-Hua LI R. steven WAGNER Primate Behavior and Ecology Program Central Washington University Department of Anthropology Central Washington University Department of Psychology Central Washington University School of Resources and Environment Anhui University School of Life Sciences Anhui Normal University Department of Biological Sciences Central Washington University

Adult male Tibetan(Macaca thibetana), Barbary(M. sylvanus), and stump-tailed macaques(M. arctoides) engage in bridging, a ritualized infant-handling behavior. Previous researchers found a bias toward the use of male infants for this behavior, but its function is debated. Explanations include three hypotheses: paternal care, mating effort, and agonistic buffering. We studied a group of habituated, provisioned Tibetan macaques to test whether adult males' affiliative relationships with females predicted their use of an infant for bridging. We also examined biases for sex, age, and individual in males' choice of bridging infant. We collected data via all occurrences, focal animal, and scan methods, from August to September 2011 at the Valley of the Wild Monkeys, China. We found that male infants were significantly preferred over females for bridging, but of three male infants in the group, only one was used by all males, while one male infant was used less often than expected. Adult males had females they were significantly more likely to be proximate to and/or to groom, but these corresponded to the mother of the bridging infant for only one male. Our results are most consistent with the agonistic buffering hypothesis: lower-ranked males used the alpha male's preferred bridging infant in an attempt to regulate their interactions with the alpha.

关键词： Agonistic buffering Affiliated infant Paternal care

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Macrophage-specific inhibition of the histone demethylase JMJD3 decreases STING and pathologic inflammation in diabetic wound repair

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Cellular & Molecular Immunology 2022年第11期19卷 1251-1262页

作者： Christopher O.Audu William J.Melvin Amrita D.Joshi Sonya J.Wolf Jadie Y.Moon Frank M.Davis Emily C.Barrett Kevin D.Mangum Hongping Deng Xianying Xing Rachel Wasikowski Lam C.Tsoi Sriganesh B.Sharma Tyler M.bauer James Shadiow Matthew A.Corriere Andrea TObi steven LKunkel Benjamin Levi Bethany BMoore Johann EGudjonsson Andrew MSmith Katherine A.Gallagher Department of Surgery Section of Vascular SurgeryUniversity of MichiganAnn ArborMIUSA Department of Microbiology and Immunology University of MichiganAnn ArborMIUSA Department of Medicinal Chemistry University of MichiganAnn ArborMIUSA Department of Surgery Section of General SurgeryUniversity of MichiganAnn ArborMIUSA Department of Bioengineering University of IllinoisUrbana-ChampaignChampaignILUSA Department of Dermatology University of MichiganAnn ArborMIUSA Department of Pathology University of MichiganAnn ArborMIUSA Department of Surgery University of Texas Southwestern Medical CenterDallasTXUSA

Macrophage plasticity is critical for normal tissue repair following *** pathologic states such as diabetes,macrophage plasticity is impaired,and macrophages remain in a persistent proinflammatory state;however,the reasons for this are ***,using single-cell RNA sequencing of human diabetic wounds,we identified increased JMJD3 in diabetic wound macrophages,resulting in increased inflammatory gene ***,we report that in wound healing,JMJD3 directs early macrophage-mediated inflammation via JAK1,3/STAT3 ***,in the diabetic state,we found that IL-6,a cytokine increased in diabetic wound tissue at later time points post-injury,regulates JMJD3 expression in diabetic wound macrophages via the JAK1,3/STAT3 pathway and that this late increase in JMJD3 induces NFκB-mediated inflammatory gene transcription in wound macrophages via an H3K27me3 ***,RNA sequencing of wound macrophages isolated from mice with JMJD3-deficient myeloid cells(Jmjd3f/fLyz2Cre+)identified that the STING gene(Tmem173)is regulated by JMJD3 in wound *** limits inflammatory cytokine production by wound macrophages during ***,in diabetic mice,its role changes to limit wound repair and enhance *** finding is important since STING is associated with chronic inflammation,and we found STING to be elevated in human and murine diabetic wound macrophages at late time ***,we demonstrate that macrophage-specific,nanoparticle inhibition of JMJD3 in diabetic wounds significantly improves diabetic wound repair by decreasing inflammatory cytokines and *** together,this work highlights the central role of JMJD3 in tissue repair and identifies cell-specific targeting as a viable therapeutic strategy for nonhealing diabetic wounds.

关键词： wound healing epigenetics diabetes JMJD3 STING

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