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Expression and reconstitution of the bioluminescent Ca2+reporter aequorin in human embryonic stem cells,and exploration of the presence of functional IP3 and ryanodine receptors during the early stages of their differentiation into cardiomyocytes

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science China(Life sciences) 2016年第8期59卷 811-824页

作者： harvey y.s.chan Man Chun Cheung yi Gao Andrew L.Miller sarah E.Webb Division of Life science&state Key Laboratory of Molecular Neuroscience HKUSTClear Water BayHong KongChina stem Cell&Regenerative Medicine Consortium Li Ka Shing Faculty of MedicineThe University of Hong KongPokfulamHong KongChina Marine Biological Laboratory Woods Hole MA 02543USA

In order to develop a novel method of visualizing possible Ca2＋ signaling during the early differentiation of hEsCs into cardi- omyocytes and avoid some of the inherent problems associated with using fluorescent reporters, we expressed the biolumines- cent Ca2＋ reporter, apo-aequorin, in HEs2 cells and then reconstituted active holo-aequorin by incubation withf-coelenterazine. The temporal nature of the Ca2＋ signals generated by the holo-f-aequorin-expressing HEs2 cells during the earliest stages of differentiation into cardiomyocytes was then investigated. Our data show that no endogenous Ca2＋ transients （generated by re- lease from intracellular stores） were detected in 1-12-day-old cardiospheres but transients were generated in cardiospheres following stimulation with KC1 or CaC12, indicating that holo-f-aequorin was functional in these cells. Furthermore, following the addition of exogenous ATP, an inositol trisphosphate receptor （IP3R） agonist, small Ca2＋transients were generated from day 1 onward. That ATP was inducing Ca2＋ release from functional IP3Rs was demonstrated by treatment with 2-APB, a known IP3R antagonist. In contrast, following treatment with caffeine, a ryanodine receptor （RyR） agonist, a minima/Ca2＋ response was observed at day 8 of differentiation only. Thus, our data indicate that unlike RyRs, IP3Rs are present and continually functional at these early stages of cardiomyocyte differentiation.

关键词： Ca2+ signaling apo-aequorin expression bioluminescence HEs2 human embryonic stem cells hEsC-derived cardiospheres IP3 and ryanodine receptors

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Expression and reconstitution of the bioluminescent Ca2+ reporter aequorin in human embryonic stem cells, and determination of the presence of functional IP3 receptors at the earliest stages of their differentiation into cardiomyocytes

Expression and reconstitution of the bioluminescent Ca2+ rep...

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第十一届全国钙信号和细胞功能研讨会

作者： harvey y.s.chan Man Chun Cheung yi Gao Andrew L.Miller sarah E.Webb Division of Life science & state Key Laboratory of Molecular Neuroscience HKUST stem Cell & Regenerative Medicine Consortium Li Ka Shing Faculty of Medicine The University of Hong Kong

In order to develop a novel method of visualizing possible Ca signaling during the early differentiation of h EsCs into cardiomyocytes and avoid some of the inherent problems associated with using fluorescent reporters, we expressed the bioluminescent Ca reporter, apoaequorin, in HEs2 cells. Active holo-aequorin was then reconstituted in the apo-aequorinexpressing cells, by incubation with f-coelenterazine, one of a variety of prosthetic co-factors of the holo-aequorin complex. Holo-f-aequorin is a Ca-sensitive bioluminescent protein complex that emits light at 70 nm on binding with Ca and it is 9 times brighter than heterogeneous natural aequorin. Being bioluminescent, aequorin does not require excitation light;it remains in the cells that it is injected into or in the cell compartments where it is targeted to;and it has minimal buffering capacity. The temporal nature of the Ca signals generated by the holo-f-aequorin-expressing HEs2 cells during the earliest stages of differentiation into h EsC-v CMs(i.e., in cardiospheres during the first 12 days) was then investigated using a luminometer. Our data show that no endogenous Ca transients(generated by release from intracellular stores) could be detected in 1-12 day old cardiospheres but that Ca transients were generated in cardiospheres following stimulation with KCl or CaCl, indicating that holof-aequorin was functional in these cells during this developmental time-window. In addition, following the addition of exogenous ATP, an agonist of the inositol trisphosphate receptor(IP3R), Ca transients were generated from day 4 onward. That ATP was inducing Ca release from functional IP3 Rs was demonstrated by treatment with 2-APB, a known IP3 R antagonist. On the other hand, following treatment with caffeine, an agonist of the ryanodine receptor(Ry R), a minimal Ca response was observed at day 8 of differentiation only. Thus, our data indicate that sarcoplasmic reticulum-based IP3 Rs are present and functional at these e

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sARs-CoV-2 variants are associated with different clinical courses in children with MIs-C

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World Journal of Pediatrics 2024年第2期20卷 143-152页

作者： Andres F.Moreno Rojas Emelia Bainto Helen harvey Adriana H.Tremoulet Jane C.Burns Kirsten B.Dummer Department of Pediatrics Division of Pediatric CardiologyUniversity of California San Diegoand Rady Children’s Hospital3020 Children’s Way MC 5004San DiegoCA 92123USA Kawasaki Disease Research Center La JollaCAUSA Department of Pediatrics Division of Pediatric Critical CareUniversity of California San Diegoand Rady Children’s HospitalSan DiegoCAUSA Department of Pediatrics University of California San Diegoand Rady Children’s HospitalSan DiegoCAUSA

Background Recent infection with sARs‑CoV-2 in children has been associated with multisystem inflammatory syndrome in children(MIs-C).sARs‑CoV-2 has undergone different *** publications exist about specific variants and their correlation with the severity of *** This was a single-center,retrospective study including all patients admitted with MIs-C at Rady Children’s Hospital-san Diego between May 2020 and March *** epidemiologic data,including viral genomic information,were obtained from public ***,clinical presentation,laboratory values,and outcomes were obtained from electronic medical *** The analysis included 104 pediatric *** MIs-C waves were *** variants in san Diego during the first wave included clades 20A to *** the second wave,there were variants from clades 20A to C,20G,21C(Epsilon),20I(Alpha),and 20J(Gamma).The third wave had Delta strains(clades 21A,21I,and 21J),and the fourth had Omicron variants(clades 21K,21L,and 22C).MIs-C presented with similar symptoms and laboratory findings across all *** patients were admitted to the pediatric intensive care unit(PICU)(74%)and required inotropic support(63%)during the second *** of the patients required mechanical circulatory support,and only two required invasive ventilatory *** was no *** The various strains of sARs-CoV-2 triggered MIs-C with differing severities,with the second wave having a more severe clinical *** the differences in disease severity across variants were due to changes in the virus or other factors remains unknown.

关键词： Complications COVID-19 MIs-C sARs-CoV-2 variants

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Cognitive function among military veterans with sTEM occupations

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Military Medical Research 2024年第2期11卷 308-310页

作者： Justin T.McDaniel Kevin N.Hascup Erin R.Hascup Ugochukwu G.Ezigbo Amanda M.Weidhuner harvey Henson David L.Albright Dale and Deborah smith Center for Alzheimer's Research and trEatment(CARE) Southern Illinois University School of MedicineSpringfieldIL 62794USA sTEM Education Research Center Southern Illinois UniversityCarbondaleIL 62901USA Department of Political science The University of AlabamaTuscaloosaAL 35487USA

Dear Editor,There is limited research on the relationship between science,technology,engineering,and mathematics(sTEM)occupational history and cognitive function in later life,especially among military veterans,who may be at greater risk for later-life cognitive *** study examines a nationally representative sample of military veterans to address this gap in knowledge.

关键词： Alzheimer's disease Veterans science technology engineering and mathematics(sTEM) Animal fluency test Dementia

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Data-driven disturbance observer-based control: an active disturbance rejection approach

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Control Theory and Technology 2021年第1期19卷 80-93页

作者： harvey Rojas-Cubides John Cortés-Romero Jaime Arcos-Legarda Deparameno de Ingeniera Elétricay Electrónica Universidad Nacional de ColombiaAv.K30 No.45-03 Edif.453 Of.222BogoiáColombia Department of Mechatromies Engineering Universidad de San BuenaventuraBogotá.Colombia

In this paper,a data-driven method for disturbance estimation and rejection is *** proposed approach is divided into two stages:an inner stabilization loop,to set the desired reference model,together with an outer loop for disturbance estimation and *** by the active disturbance rejection control framework,the exogenous and endogenous disturbances are lumped into a lotal disturbance *** signal is estimaed using an on-line algorithm based on a data-driven predictor scheme,whose parameters are chosen Io salisfy high robustnessperformance *** above process is presented as a novel enhancement lo design a disturbance observer,w hich constitutes the main contribution of the *** addition,the control strategy is completely presented in discrete time,avoiding the use of discretization methods for its digital *** a case study,the voltage control of a DC-DC synchronous buck converter aflected by disturbances in the input voltage and the load is ***,experimental results that validate the proposed stralegy and some comparisons with the classical disturbance observer-based control are presented.

关键词： Active disturbance rejection control Data-driven disturbance observer Disturbance observer-based control signal prediction

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Mechanical properties evaluation for thin film/substrate material systems

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Theoretical & Applied Mechanics Letters 2018年第1期8卷 1-2页

作者： simon s.Wang Zhanwei Liu Christopher M.harvey Department of Aeronautical and Automotive Engineering Loughborough University school of Aerospace Engineering Beijing Institute of Technology

Thin film/substrate material systems have been widely used in various industries such as aerospace,automotive,marine and *** example is thermal barrier coatings(TBCs):TBCs are extensively applied to metallic components on gas turbine engines in order to protect them from high and prolonged heat loads,and they play an essential role in the aircraft *** example is laminated polymer matrix composites(PMCs),reinforced with fibers such as carbon,glass or aramid:These materials have been extensively employed in a variety of applications due to their superior properties and the improving ease and cost-effectiveness of fabricating *** evaluation of mechanical properties for thin film/substrate material systems is imperative for more efficient use of these material systems and further improvements.

关键词： TBCs PMCs various industries

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Highly efficient surface-emitting semiconductor lasers exploiting quasi-crystalline distributed feedback photonic patterns

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Light(science & Applications) 2020年第1期9卷 1498-1508页

作者： simone Biasco Andrea Ciavatti Lianhe Li AGiles Davies Edmund H.Linfield harvey Beere David Ritchie Miriam s.Vitiello NEsT CNR-Istituto Nanoscienze and Scuola Normale SuperiorePiazza San Silvestro 1256127 PisaItaly school of Electronic and Electrical Engineering University of LeedsLeeds LS29JTUK Cavendish Laboratory University of CambridgeCambridge CB30HEUK

Quasi-crystal distributed feedback lasers do not require any form of mirror cavity to amplify and extract *** implemented on the top surface of a semiconductor laser,a quasi-crystal pattern can be used to tune both the radiation feedback and the extraction of highly radiative and high-quality-factor optical modes that do not have a defined symmetric or anti-symmetric ***,this methodology offers the possibility to achieve efficient emission,combined with tailored spectra and controlled beam ***,we apply this concept to a onedimensional quantum cascade wire *** lithographically patterning a series of air slits with different widths,following the Octonacci sequence,on the top metal layer of a double-metal quantum cascade laser operating at THz frequencies,we can vary the emission from single-frequency-mode to multimode over a 530-GHz bandwidth,achieving a maximum peak optical power of 240mW(190 mW)in multimode(single-frequency-mode)lasers,with record slope efficiencies for multimode surface-emitting disordered THz lasers up to ≈570 mW/A at 78 K and ≈720 mW/A at 20 K and wall-plug efficiencies of η≈1%.

关键词： lasers surface quantum

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Global sensitivity analysis of a single-cell HBV model for viral dynamics in the liver

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Infectious Disease Modelling 2021年第1期6卷 1220-1235页

作者： Md Afsar Ali s.A.Means harvey Ho Jane Heffernan york University Canada Massey University New Zealand The University of Auckland New Zealand

The predictive accuracy of mathematical models representing anything ranging from the meteorological to the biological system profoundly depends on the quality of model parameters derived from experimental ***,robust sensitivity analysis(sA)of these critical model parameters aids in sifting the influential from the negligible out of typically vast parameter regimes,thus illuminating key components of the system under *** here move beyond traditional local sensitivity analysis to the adoption of global sA *** rank correlation coefficient(PRCC)based on Latin hypercube sampling is compared with the variance-based sobol *** selected for this sA investigation an infection model for the hepatitis-B virus(HBV)that describes infection dynamics and clearance of HBV in the liver[Murray&Goyal,2015].The model tracks viral particles such as the tenacious and nearly ineradicable covalently closed circular DNA(cccDNA)embedded in infected nuclei and an HBV protein known as *** application of these sA methods to the HBV model illuminates,especially over time,the quantitative relationships between cccDNA synthesis rate and p36 synthesis and *** results reinforce previous observations that the viral protein,p36,is by far the most influential factor for cccDNA ***,both methods are capable of finding crucial parameters of the *** the sobol method is independent of model structure(e.g.,linearity and monotonicity)and well suited for sA,our results ensure that LHs-PRCC suffices for sA of a non-linear model if it is monotonic.

关键词： Latin hypercube sampling Partial rank correlation coefficient(PRCC) sobol method HBV Liver

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Regulating Wnt signaling： a strategy to prevent neurodegeneration and induce regeneration

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Journal of Molecular Cell Biology 2014年第1期8卷 1-2页

作者： Guest Editors Kirsten harvey Bianca Marchetti Department of Pharmaco-logy UCL School of PharmacyUniversity College LondonLondon WCIN lAX UK Department of Clinical andMolecular Biomedicine Phar-macology Section MedicalSchool University of Catania95125 Catania Italy 0A51 Institute for Researchand Care on Mental Retarda-tion and Brain Aging (IRCCs) Neurapharmacology Section94018 Tmina Italy

During the past three decades, Wingtess/Int （Wnt）signaling has emerged as an essential regu{ator crucial for neuronal development and maintenance （Inestrosa and Arenas, 201_0）. In addition, Wnt signal- ing was recently shown to be involved in the regula- tion of synaptic function and plasticity, which is critical for learning and memory （Oliva et aL, 2013）. Deregulation of Wnt signaling has been proposed as a key contributor to the pathogenesis of neurode- generative disorders including Alzheimer＇s disease （AD） and Parkinson＇s disease （PD）. This increasing knowledge of the specific roles of Wnt signaling cascades during different stages of life has suggested innovative therapeutic strategies for the treatment of neurodegenerative diseases.

关键词： Wnt 神经元退行性病变信号神经退行性疾病阿尔茨海默氏病再生诱发

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Evolution of breast cancer therapeutics: Breast tumour kinase's role in breast cancer and hope for breast tumour kinase targeted therapy

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World Journal of Clinical Oncology 2014年第3期5卷 299-310页

作者： Haroon A Hussain Amanda J harvey Brunel Institute for Cancer Genetics and Pharmacogenomics Biosciences Brunel University (London)

There have been significant improvements in the detection and treatment of breast cancer in recent decades. However, there is still a need to develop more effective therapeutic techniques that are patient specific with reduced toxicity leading to further increases in patients' overall survival; the ongoing progress in understanding recurrence, resistant and spread also needs to be maintained. Better understanding of breast cancer pathology, molecular biology and progression as well as identification of some of the underlying factors involved in breast cancer tumourgenesis and metastasis has led to the identification of novel therapeutic targets. Over a number of years interest has risen in breast tumour kinase(Brk) also known as protein tyrosine kinase 6; the research field has grown and Brk has been described as a desirable therapeutic target in relation to tyrosine kinase inhibition as well as disruption of its kinase independent activity. This review will outline the current "state of play" with respect to targeted therapy for breast cancer, as well as discussing Brk's role in the processes underlying tumour development and metas-tasis and its potential as a therapeutic target in breast cancer.

关键词： Breast tumour kinase Protein tyrosine kinase 6 Breast neoplasms Targeted molecular therapy Intracellular signaling peptides and proteins Protein kinase inhibitors

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