RESUMEN
Improved functional ability and physical activity are strongly associated with a broad range of positive health outcomes including reduced risk of hospital readmission. This study presents an algorithm for detecting ambulations from time-resolved step counts gathered from remote monitoring of patients receiving hospital care in their homes. It examines the statistical power of these ambulations in predicting hospital readmission. A diverse demographic cohort of 233 patients of age 70.5±16.8 years are evaluated in a retrospective analysis. Eleven statistical features are derived from raw time series data, and their F-statistics are assessed in discriminating between patients who were and were not readmitted within 30 days of discharge. Using these features, logistic regression models are trained to predict readmission. The results show that the fraction of days with at least one ambulation was the strongest feature, with an F-statistic of 17.2. The models demonstrate AUROC performances of 0.741, 0.766 and 0.769 using stratified 5-fold train-test splits in all included patients (n=233), congestive heart failure (CHF, n=105) and non-CHF (n=128) patient subgroups, respectively. This study suggests that patient ambulation metrics derived from wearable sensors can offer powerful predictors of adverse clinical outcomes such as hospital readmission, even in the absence of other features such as physiological vital signs.Index Terms-readmission, ambulation, step count, heart failure, physical activity, regression, actigraphy, accelerometer.
Asunto(s)
Insuficiencia Cardíaca , Readmisión del Paciente , Anciano , Anciano de 80 o más Años , Humanos , Modelos Logísticos , Persona de Mediana Edad , Estudios Retrospectivos , CaminataRESUMEN
Maquettes are man-made cofactor-binding oxidoreductases designed from first principles with minimal reference to natural protein sequences. Here we focus on water-soluble maquettes designed and engineered to perform diffusive electron transport of the kind typically carried out by cytochromes, ferredoxins and flavodoxins and other small proteins in photosynthetic and respiratory energy conversion and oxido-reductive metabolism. Our designs were tested by analysis of electron transfer between heme maquettes and the well-known natural electron transporter, cytochrome c. Electron-transfer kinetics were measured from seconds to milliseconds by stopped-flow, while sub-millisecond resolution was achieved through laser photolysis of the carbon monoxide maquette heme complex. These measurements demonstrate electron transfer from the maquette to cytochrome c, reproducing the timescales and charge complementarity modulation observed in natural systems. The ionic strength dependence of inter-protein electron transfer from 9.7×10(6) M(-1) s(-1) to 1.2×10(9) M(-1) s(-1) follows a simple Debye-Hückel model for attraction between +8 net charged oxidized cytochrome c and -19 net charged heme maquette, with no indication of significant protein dipole moment steering. Successfully recreating essential components of energy conversion and downstream metabolism in man-made proteins holds promise for in vivo clinical intervention and for the production of fuel or other industrial products. This article is part of a Special Issue entitled Biodesign for Bioenergetics--the design and engineering of electronic transfer cofactors, proteins and protein networks, edited by Ronald L. Koder and J.L. Ross Anderson.
Asunto(s)
Citocromos c/química , Proteínas del Complejo de Cadena de Transporte de Electrón/química , Ingeniería de Proteínas/métodos , Secuencia de Aminoácidos , Citocromos c/genética , Citocromos c/metabolismo , Difusión , Transporte de Electrón/genética , Proteínas del Complejo de Cadena de Transporte de Electrón/genética , Proteínas del Complejo de Cadena de Transporte de Electrón/metabolismo , Hemo/metabolismo , Humanos , Cinética , Modelos Moleculares , Datos de Secuencia Molecular , Oxidación-Reducción , Fotólisis , Estructura Terciaria de Proteína , Homología de Secuencia de AminoácidoRESUMEN
Here we describe the design, Escherichia coli expression and characterization of a simplified, adaptable and functionally transparent single chain 4-α-helix transmembrane protein frame that binds multiple heme and light activatable porphyrins. Such man-made cofactor-binding oxidoreductases, designed from first principles with minimal reference to natural protein sequences, are known as maquettes. This design is an adaptable frame aiming to uncover core engineering principles governing bioenergetic transmembrane electron-transfer function and recapitulate protein archetypes proposed to represent the origins of photosynthesis. This article is part of a Special Issue entitled Biodesign for Bioenergetics--the design and engineering of electronic transfer cofactors, proteins and protein networks, edited by Ronald L. Koder and J.L. Ross Anderson.
Asunto(s)
Proteínas del Complejo de Cadena de Transporte de Electrón/química , Metabolismo Energético , Proteínas de la Membrana/química , Ingeniería de Proteínas/métodos , Secuencia de Aminoácidos , Proteínas del Complejo de Cadena de Transporte de Electrón/genética , Proteínas del Complejo de Cadena de Transporte de Electrón/metabolismo , Metabolismo Energético/genética , Escherichia coli , Hemo/química , Hemo/metabolismo , Humanos , Proteínas de la Membrana/genética , Proteínas de la Membrana/metabolismo , Datos de Secuencia Molecular , Fotosíntesis , Unión Proteica , Pliegue de Proteína , Estructura Terciaria de Proteína , Homología de Secuencia de AminoácidoRESUMEN
The first principles design of manmade redox-protein maquettes is used to clarify the physical/chemical engineering supporting the mechanisms of natural enzymes with a view to recapitulate and surpass natural performance. Herein, we use intein-based protein semisynthesis to pair a synthetic naphthoquinone amino acid (Naq) with histidine-ligated photoactive metal-tetrapyrrole cofactors, creating a 100â µs photochemical charge separation unit akin to photosynthetic reaction centers. By using propargyl groups to protect the redox-active para-quinone during synthesis and assembly while permitting selective activation, we gain the ability to employ the quinone amino acid redox cofactor with the full set of natural amino acids in protein design. Direct anchoring of quinone to the protein backbone permits secure and adaptable control of intraprotein electron-tunneling distances and rates.
Asunto(s)
Aminoácidos/química , Luz , Naftoquinonas/química , Proteínas del Complejo del Centro de Reacción Fotosintética/química , Aminoácidos/síntesis química , Transporte de Electrón/efectos de la radiación , Inteínas , Modelos Moleculares , Estructura Molecular , Naftoquinonas/síntesis química , Procesos Fotoquímicos/efectos de la radiaciónRESUMEN
Emulating functions of natural enzymes in man-made constructs has proven challenging. Here we describe a man-made protein platform that reproduces many of the diverse functions of natural oxidoreductases without importing the complex and obscure interactions common to natural proteins. Our design is founded on an elementary, structurally stable 4-α-helix protein monomer with a minimalist interior malleable enough to accommodate various light- and redox-active cofactors and with an exterior tolerating extensive charge patterning for modulation of redox cofactor potentials and environmental interactions. Despite its modest size, the construct offers several independent domains for functional engineering that targets diverse natural activities, including dioxygen binding and superoxide and peroxide generation, interprotein electron transfer to natural cytochrome c and light-activated intraprotein energy transfer and charge separation approximating the core reactions of photosynthesis, cryptochrome and photolyase. The highly stable, readily expressible and biocompatible characteristics of these open-ended designs promise development of practical in vitro and in vivo applications.
Asunto(s)
Oxidorreductasas/metabolismo , Proteínas/química , Hemo/química , Hemo/metabolismo , Modelos Moleculares , Estructura Molecular , Resonancia Magnética Nuclear Biomolecular , Oxidorreductasas/química , Unión Proteica , Conformación Proteica , Ingeniería de Proteínas/métodosRESUMEN
The study of natural enzymes is complicated by the fact that only the most recent evolutionary progression can be observed. In particular, natural oxidoreductases stand out as profoundly complex proteins in which the molecular roots of function, structure and biological integration are collectively intertwined and individually obscured. In the present paper, we describe our experimental approach that removes many of these often bewildering complexities to identify in simple terms the necessary and sufficient requirements for oxidoreductase function. Ours is a synthetic biology approach that focuses on from-scratch construction of protein maquettes designed principally to promote or suppress biologically relevant oxidations and reductions. The approach avoids mimicry and divorces the commonly made and almost certainly false ascription of atomistically detailed functionally unique roles to a particular protein primary sequence, to gain a new freedom to explore protein-based enzyme function. Maquette design and construction methods make use of iterative steps, retraceable when necessary, to successfully develop a protein family of sturdy and versatile single-chain three- and four-α-helical structural platforms readily expressible in bacteria. Internally, they prove malleable enough to incorporate in prescribed positions most natural redox cofactors and many more simplified synthetic analogues. External polarity, charge-patterning and chemical linkers direct maquettes to functional assembly in membranes, on nanostructured titania, and to organize on selected planar surfaces and materials. These protein maquettes engage in light harvesting and energy transfer, in photochemical charge separation and electron transfer, in stable dioxygen binding and in simple oxidative chemistry that is the basis of multi-electron oxidative and reductive catalysis.