Lessons Learned from a Multi-Site, Team-Based Serious Illness Care Program Implementation at an Academic Medical Center.

Patients with serious illness benefit from conversations to share prognosis and explore goals and values. To address this, we implemented Ariadne Labs' Serious Illness Care Program (SICP) at Stanford Health Care. Improve quantity, timing, and quality of serious illness conversations.

Considerations in the reliability and fairness audits of predictive models for advance care planning.

Multiple reporting guidelines for artificial intelligence (AI) models in healthcare recommend that models be audited for reliability and fairness. However, there is a gap of operational guidance for performing reliability and fairness audits in practice. Following guideline recommendations, we conducted a reliability audit of two models based on model performance and calibration as well as a fairness audit based on summary statistics, subgroup performance and subgroup calibration.

First principles design of a core bioenergetic transmembrane electron-transfer protein.

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.

Rheostat re-wired: alternative hypotheses for the control of thioredoxin reduction potentials.

Thioredoxins are small soluble proteins that contain a redox-active disulfide (CXXC). These disulfides are tuned to oxidizing or reducing potentials depending on the function of the thioredoxin within the cell. The mechanism by which the potential is tuned has been controversial, with two main hypotheses: first, that redox potential (Em) is specifically governed by a molecular 'rheostat'-the XX amino acids, which influence the Cys pKa values, and thereby, Em; and second, the overall thermodynamics of protein folding stability regulates the potential.

Enzymatic and cryoreduction EPR studies of the hydroxylation of methylated N(ω)-hydroxy-L-arginine analogues by nitric oxide synthase from Geobacillus stearothermophilus.

Nitric oxide synthase (NOS) catalyzes the conversion of L-arginine to L-citrulline and NO in a two-step process involving the intermediate N(ω)-hydroxy-L-arginine (NHA). It was shown that Cpd I is the oxygenating species for L-arginine; the hydroperoxo ferric intermediate is the reactive intermediate with NHA. Methylation of the N(ω)-OH and N(ω)-H of NHA significantly inhibits the conversion of NHA into NO and L-citrulline by mammalian NOS.

Engineering oxidoreductases: maquette proteins designed from scratch.

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.

Quinone and non-quinone redox couples in Complex III.

The Q cycle mechanism proposed by Peter Mitchell in the 1970's explicitly considered the modification of ubiquinone two-electron redox properties upon binding to Complex III to match the thermodynamics of the other single-electron redox cofactors in the complex, and guide electron transfer to support the generation of a proton electro-chemical gradient across native membranes. A better understanding of the engineering of Complex III is coming from a now moderately well defined thermodynamic description of the redox components as a function of pH, including the Qi/heme b(H) cluster. The redox properties of the most obscure component, Qo, is finally beginning to be resolved.

Breaking the Q-cycle: finding new ways to study Qo through thermodynamic manipulations.

Thirty years ago, Peter Mitchell won the Nobel Prize for proposing how electrical and proton gradients across bioenergetic membranes were the energy coupling intermediate between photosynthetic and respiratory electron transfer and cellular activities that include ATP production. A high point of his thinking was the development of the Q-cycle model that advanced our understanding of cytochrome bc (1). While the principle tenets of his Q-cycle still hold true today, Mitchell did not explain the specific mechanism that allows the Qo site to perform this Q-cycle efficiently without undue energy loss.

Direct electrochemical analyses of a thermophilic thioredoxin reductase: interplay between conformational change and redox chemistry.

Thioredoxin reductases (TrxRs) are flavin-containing dithioloxidoreductases that couple reduction equivalents from the soluble NAD(P)H pool to the soluble protein thioredoxin (Trx). Previous crystallographic studies of the Escherichia coli enzyme ( ecTrxR) have shown that low molecular weight TrxRs can adopt two distinct conformations: the first (FO) is required for the oxidation of the flavin cofactor and the generation of reduced Trx; the second (FR) is adopted for the reduction of the flavin by NAD(P)H. Here, protein electrochemistry has been used to interrogate the equilibrium between the oxidized and reduced conformations of the ecTrxR and a novel, low molecular weight TrxR from the thermophilic archaeon Thermoplasma acidophilum ( taTrxR) that is characterized structurally and biochemically in the accompanying paper [Hernandez et al.

Distance metrics for heme protein electron tunneling.

There is no doubt that distance is the principal parameter that sets the order of magnitude for electron-tunneling rates in proteins. However, there continue to be varying ways to measure electron-tunneling distances in proteins. This distance uncertainty blurs the issue of whether the intervening protein medium has been naturally selected to speed or slow any particular electron-tunneling reaction.

Electron tunneling chains of mitochondria.

The single, simple concept that natural selection adjusts distances between redox cofactors goes a long way towards encompassing natural electron transfer protein design. Distances are short or long as required to direct or insulate promiscuously tunneling single electrons. Along a chain, distances are usually 14 A or less.

Chemogenomic profiling on a genome-wide scale using reverse-engineered gene networks.

A major challenge in drug discovery is to distinguish the molecular targets of a bioactive compound from the hundreds to thousands of additional gene products that respond indirectly to changes in the activity of the targets. Here, we present an integrated computational-experimental approach for computing the likelihood that gene products and associated pathways are targets of a compound. This is achieved by filtering the mRNA expression profile of compound-exposed cells using a reverse-engineered model of the cell's gene regulatory network.

A distinctive electrocatalytic response from the cytochrome c peroxidase of nitrosomonas europaea.

Here the cytochrome c peroxidase (CcP) from Nitrosomonas europaea is examined using the technique of catalytic protein film voltammetry. Submonolayers of the bacterial diheme enzyme at a pyrolytic graphite edge electrode give catalytic, reductive signals in the presence of the substrate hydrogen peroxide. The resulting waveshapes indicate that CcP is bound non-covalently in a highly active configuration.

Endemic incidence of infections caused by Mycobacterium kansasii in the Karviná district in 1968-1995 (analysis of epidemiological data--review).

The Karviná district (northwestern part of North Moravia and Silesia) is typical by its industrial character, above all mining of black coal which is coked and which led to the devastation of the countryside. The ratio of light industry is low. In a small area (347 km2) there is a population of 824 inhabitants per km2.

[Acute viral respiratory diseases as a cause of work disability in miners during an inter-epidemic period].

During the interepidemic period between October 1 and November 20 1987 96 miners from a coal mine were examined who suffered from diseases of the upper airways which caused their work incapacity. In paired sera the rise of antibodies against M. pneumoniae and respiratory viruses were examined (adeno, influenza virus A and B, Coxsackie A21, corona 229E and OC43, parainfluenza type 1, 2 and 3, rhino type 13 and 44, RS).

Mycobacterium kansasii infection in an endemic area of Czechoslovakia.

Over the period 1968-1979, Mycobacterium kansasii was isolated from a total of 297 persons residing in the district of Karviná, North-Moravian Region, Czechoslovakia. Repeated M. kansasii isolations were recorded in 153 patients with clinical symptomatology of chronic respiratory disorders, single isolations in another 144 persons (121 isolations from sputum and/or laryngeal swabs, 22 from urine, and one from the aspirate of a lymph node with tuberculoid granuloma).

Mass occurrence of Yersinia enterocolitica in two establishments of collective care of children.

Mass occurrence of intestinal disorders caused by Yersinia enterocolitica simultaneously in two groups of children of under-school age is reported. The question of the source of infection and the routes of transfer are discussed and the various forms pointed out which Y. e.