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431.
Igor Jurak Michael Hackenberg Ju Youn Kim Jean M. Pesola Roger D. Everett Chris M. Preston Angus C. Wilson Donald M. Coen 《Journal of virology》2014,88(4):2337-2339
To facilitate studies of herpes simplex virus 1 latency, cell culture models of quiescent or latent infection have been developed. Using deep sequencing, we analyzed the expression of viral microRNAs (miRNAs) in two models employing human fibroblasts and one using rat neurons. In all cases, the expression patterns differed from that in productively infected cells, with the rat neuron pattern most closely resembling that found in latently infected human or mouse ganglia in vivo. 相似文献
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The maintenance of transbilayer distribution of phospholipids is crucial for proper cell function. Intramembrane transport of lipids is mediated by three activities termed floppases, flippases, and scramblases. Members of the ATP-binding cassette transporter family and P-type ATPase superfamily have been implicated in the translocation of lipids. The importance of these activities is exemplified by several severe human inherited disorders that are caused by defects in intramembranous transport of lipids. In order to elucidate the molecular mechanisms that underlie these disorders, the combination of in vivo, biochemical, and structural analyses on intramembrane transporters is crucial. 相似文献
435.
Shelby Giza Jorge A. MojicaSantiago Maddalena Parafati Legrand K. Malany Don Platt Christine E. Schmidt Paul M. Coen Siobhan Malany 《Aging cell》2022,21(7)
Microphysiological systems (MPS), also referred to as tissue chips, incorporating 3D skeletal myobundles are a novel approach for physiological and pharmacological studies to uncover new medical treatments for sarcopenia. We characterize a MPS in which engineered skeletal muscle myobundles derived from donor‐specific satellite cells that model aged phenotypes are encapsulated in a perfused tissue chip platform containing platinum electrodes. Our myobundles were derived from CD56+ myogenic cells obtained via percutaneous biopsy of the vastus lateralis from adults phenotyped by age and physical activity. Following 17 days differentiation including 5 days of a 3 V, 2 Hz electrical stimulation regime, the myobundles exhibited fused myotube alignment and upregulation of myogenic, myofiber assembly, signaling and contractile genes as demonstrated by gene array profiling and localization of key components of the sarcomere. Our results demonstrate that myobundles derived from the young, active (YA) group showed high intensity immunofluorescent staining of α‐actinin proteins and responded to electrical stimuli with a ~1 μm displacement magnitude compared with non‐stimulated myobundles. Myobundles derived from older sedentary group (OS) did not display a synchronous contraction response. Hypertrophic potential is increased in YA‐derived myobundles in response to stimulation as shown by upregulation of insulin growth factor (IGF‐1), α‐actinin (ACTN3, ACTA1) and fast twitch troponin protein (TNNI2) compared with OS‐derived myobundles. Our MPS mimics disease states of muscle decline and thus provides an aged system and experimental platform to investigate electrical stimulation mimicking exercise regimes and may be adapted to long duration studies of compound efficacy and toxicity for therapeutic evaluation against sarcopenia. 相似文献
436.
Fine mapping and molecular cloning of mutations in the herpes simplex virus DNA polymerase locus. 总被引:13,自引:27,他引:13 下载免费PDF全文
D M Coen D P Aschman P T Gelep M J Retondo S K Weller P A Schaffer 《Journal of virology》1984,49(1):236-247
Mutations in five phenotypically distinct mutants derived from herpes simplex virus type 1 strain KOS which lie in or near the herpes simplex virus DNA polymerase (pol) locus have been fine mapped with the aid of cloned fragments of mutant and wild-type viral DNAs to distinct restriction fragments of 1.1 kilobase pairs (kbp) or less. DNA sequences containing a mutation or mutations conferring resistance to the antiviral drugs phosphonoacetic acid, acyclovir, and arabinosyladenine of pol mutant PAAr5 have been cloned as a 27-kbp Bg+II fragment in Escherichia coli. These drug resistance markers have been mapped more finely in marker transfer experiments to a 1.1-kbp fragment (coordinates 0.427 to 0.434). In intratypic marker rescue experiments, temperature-sensitive (ts), phosphonoacetic acid resistance, and acyclovir resistance markers of pol mutant tsD9 were mapped to a 0.8-kbp fragment at the left end of the EcoRI M fragment (coordinates 0.422 to 0.427). The ts mutation of pol mutant tsC4 maps within a 0.3-kbp sequence (coordinates 0.420 to 0.422), whereas that of tsC7 lies within the 1.1-kbp fragment immediately to the left (coordinates 0.413 to 0.420). tsC4 displays the novel phenotype of hypersensitivity to phosphonoacetic acid; however, the phosphonoacetic acid hypersensitivity phenotype is almost certainly not due to the mutation(s) conferring temperature sensitivity. The ts mutation of mutant tsN20--which does not affect DNA polymerase activity--maps to a 0.5-kbp fragment at the right-hand end of the EcoRI M fragment (coordinates 0.445 to 0.448). The mapping of the mutations in these five mutants further defines the limits of the pol locus and separates mutations differentially affecting catalytic functions of the polymerase. 相似文献
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We present an empirical model of Arabidopsis (Arabidopsis thaliana), intended as a framework for quantitative understanding of plant development. The model simulates and realistically visualizes development of aerial parts of the plant from seedling to maturity. It integrates thousands of measurements, taken from several plants at frequent time intervals. These data are used to infer growth curves, allometric relations, and progression of shapes over time, which are incorporated into the final three-dimensional model. Through the process of model construction, we identify the key attributes required to characterize the development of Arabidopsis plant form over time. The model provides a basis for integrating experimental data and constructing mechanistic models. 相似文献
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