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BackgroundThis study aimed to capture the implementation process of the ALIGN Study, (An individualized Pain Plan with Patient and Provider Access for Emergency Department care of Sickle Cell Disease). ALIGN aimed to embed Individualized Pain Plans in the electronic health record (E-IPP) and provide access to the plan for both adult patients with sickle cell disease (SCD) and emergency department providers when a person with SCD comes to the emergency department in vaso-occlusive crises.MethodsSemi-structured interviews were conducted with research teams from the 8 participating sites from the ALIGN study. Seventeen participants (principal investigators and study coordinators) shared their perspectives about the implementation of ALIGN in their sites. Data were analyzed in three phases using open coding steps adapted from grounded theory and qualitative content analysis.ResultsA total of seven overarching themes were identified: (1) the E-IPP structure (location and upkeep) and collaboration with the informatics team, (2) the role of ED champion, (3) the role of research coordinators, (4) research team communication, and communication between research team and clinical team, (5) challenges with the study protocol, (6) provider feedback: addressing over-utilizers, patient mistrust, and the positive feedback about the intervention, and (7) COVID-19 and its effects on study implementation.ConclusionsFindings from this study contribute to learning how to implement E-IPPs for adult patients with SCD in ED. The study findings highlight the importance of early engagement with different team members, a champion from the emergency department, study coordinators with different skills and enhancement of communication and trust among team members. Further recommendations are outlined for hospitals aiming to implement E-IPP for patients with SCD in ED.

The rotational mobilities of small solute molecules encapsulated in tetramethyyl orthosilicate sol-gels are investigated by EPR spectroscopy of encapsulated nitroxide probes and high-resolution NMR spectroscopic measurements. The electrostatic attraction between cationic probes and the negatively charged sol-gels walls causes the positively charged probes to be more effectively immobilized.

The first observations of photoinitiated interprotein electron transfer (ET) within sol-gels are described. The initial step in using sol-gel encapsulation as a tool for temporally isolating the different dynamic processes coupled to ET in protein-protein complexes is represented.

Research was conducted to examine the binding affinity and electron transfer rate constants for the complex of hemoglobin (Hb) and cytochrome b5 (b5) by triple quenching titrations of mixed-metal Hb hybrids and of fully substituted Zn-mesoporphyrin by b5. Experiments with the hybrids allow the selective monitoring of the reaction of a single chain type with Fe3+b5 and T-state Hbs. Results are consistent with a docking model in which b5 reacts with the alpha-chains by binding at a site spanning the alpha-1-beta-2 dimer interface.

Mixed-metal, (M, Fe) hemoglobin hybrids, with Fe in one type of subunit and M = Zn or Mg in the other type, deposited in clear poly(vinyl alcohol) films, showed inter-subunit electron transfer (ET) electron-nuclear tunneling down to cyrogenic temperatures. This is the first protein system other than photosynthetic systems to display such behavior. Some of the hybrids presented a sharp rise in the ET rate constant up to 200 K.

Flash photolysis triplet-quenching and electron-transfer measurements at pH values between 6 and 7.5 were done to study the formation of the (Zn-myoglobin, cytochrome b5) complex produced from the reaction of Zn deuteroporphyrin-substituted myoglobin (ZnDMb) with cytochrome b5 (Feb5). The results obtained confirmed the importance of global and local electrostatics in the regulation of protein-protein recognition and reactivity.

A comparative three-pulse electron spin echo envelope modulation study is performed on the diiron(II) centers of the two MMOHred, N3Hrred, N3Ribonucleotide reductase, N3Rubrerythin (N3Rrred) and N3-'chopped' Rubrerythin (N3CRrred), which lacks the 39-residue C-terminal FeS4 domain. Data indicate the significant role of histidyl ligands to the cluster in determining the electronic structure of the functionally important diferrous state of the proteins.

The kinetics of the electron transfer (ET) in protein-protein complexes depend on the extent and stoichiometry binding, and the interfacial dynamics of docking. The ET within protein-protein complexes produces noncovalent complexes. Cytochrome c peroxidase (CcP) forms a multidomain 2:1 binding complex of cytochrome c. The reduction of the Trp radical by electron transfer occurs under conditions of slow equilibration of CcP-II(sub h) and CcP-II(sub r). The reduction is a two-step process and occurs at the high affinity domain.

Sickle cell disease (SCD) is a genetic disorder predominantly affecting people of African descent and is associated with significant morbidity and mortality. To improve SCD outcomes, the National Heart Lung and Blood Institute funded eight centers to participate in the SCD Implementation Consortium. Sites were required to each recruit 300 individuals with SCD, over 20 months. We aim to describe recruitment strategies and challenges encountered. Participants aged 15–45 years with confirmed diagnosis of SCD were eligible. Descriptive statistics were used to analyze the effectiveness of each recruitment strategy. A total of 2432 participants were recruited. Majority (95.3%) were African American. Successful strategies were recruitment from clinics (68.1%) and affiliated sites (15.6%). Recruitment at community events, emergency departments and pain centers had the lowest yield. Challenges included saturation of strategies and time constraints. Effective recruitment of participants in multi-site studies requires multiple strategies to achieve adequate sample sizes. [ABSTRACT FROM AUTHOR]

A strategy is described by which reactive binding of a weakly bound, 'dynamically docked (DD)', complex without a known structure is strengthened electrostatically through optimized placement of surface charges, and its utility in modulating complex formation between myoglobin (Mb) and cytochrome [b.sub.5] ([b.sub.5]) is described. The measurements have shown that the presence of fast photoinduced singlet-electron transfer (ET) across the protein-protein interface.

A myoglobin (Mb) triple mutant (D44K/D60K/E85K, Mb(+6)) substituted with Zn-deuteroporphyrin is prepared and cytochrome [b.sub.5] (cyt [b.sub.5]) binding and electron transfer (ET) quenching of the [super 3]ZnMb(+6) triplet state is monitored. The mutations have created the conversion from 'Dynamic Docking' (DD) to simple docking 'simple docking' (SD) energy landscapes.

The development of simple hemoglobin (Hb) hybrid system to explore dynamic control of an electron transfer (ET) photocycle within a predocked protein-protein complex is reported. The results have shown how the conformational routes by which a hybrid progresses through a photocycle are dissimilar in different dynamic regimes.

The article studies the dynamic docking of cytochrome [b.sub.5] in reaction with its physiological partners, myoglobin (Mb) and [alpha]-hemoglobin (Hb), the two most commonly used electron transfer (ET) complexes. The complexes are found to form several heme-neutralization squares, in addition to the binding of the electron-transfer-reactive configurations.

The EPR and ENDOR studies are performed on the peroxoferriheme primary cryoreduction products of human HbO2, isolated alphaO2 and betaO2 chains, under a wide range of conditions involving the addition of glycols, sugars and IHP. The result shows that oxy alpha and beta chains that are isolated in the Hb tetramer, display two major conformational substates of the oxyheme center.

A new theoretical approach, which mathematically combines PATHWAY donor/acceptor coupling calculations with Poisson-Boltzmann-based electrostatics, is presented. This Brownian Dynamics docking profiles and PATHWAYS calculations allow visualizing the microscopic basis of Dynamic Docking.

A broad study of the effect of neutralizing the two negative charges of the Mb propionates on the interaction and electron transfer (ET) between horse Mb and bovine cyt b5, through use of Zn-substituted Mb(ZnMb, 1) is presented. The new model describes situations in which tight binding does not correlate with efficient ET reactivity, and explains how it is possible to modulate reactivity without changing affinity.

We present the first steps in applying transverse relaxation-optimized spectroscopy (TROSY) techniques to the study of allosterism. Each [Beta]-chain of the hemoglobin (Hb) tetramer has 17 valine residues. We have [sup.15]N-labeled the [Beta]-chain Val residues and detected 16 of the 17 [sup.1]H-[sup.15]N correlation peaks for [Beta]-chain Val of the R state CO-Hb structure by using the TROSY technique. Sequence-specific assignments are suggested, based mainly on analysis of the [sup.1]H pseudocontact-shift increments produced by oxidizing the diamagnetic R state HbCO to the paramagnetic R state fluoromet form. When possible, we support these assignments with sequential nuclear Overhauser effect (NOE) information obtained from a two-dimensional [[sup.1]H, [sup.1]H]-NOESY-TROSY experiment (NOESY, NOE spectroscopy). We have induced further the R-T conformational change by adding the allosteric effector, inositol hexaphosphate, to the fluoromet-Hb sample. This change induces substantial increments in the [sup.1]H and [sup.15]N chemical shifts, and we discuss the implication of these findings in the context of the tentative sequence assignments. These preliminary results suggest that amide nitrogen and amide proton chemical shifts in a selectively labeled sample are site-specific probes for monitoring the allosteric response of the ensemble-averaged solution structure of Hb. More important, the chemical-shift dispersion obtained is adequate to permit a complete assignment of the backbone [sup.15]N/[sup.13]C resonances upon nonselective labeling.

An effort is made to show that it is possible to acquire high-resolution, solution NMR measurements of sol-gel (SG)-encapsulated proteins and also that extensive studies are possible with labeled proteins. The combination of SG-encapsulation and NMR spectroscopy has provided a structural data on intermediates formed by ligand photolysis and snapshots of conformation changes during ligand rebinding.

Studies based on mixed-metal hemoglobin(Hb) hybrids provide an ideal system with electron transfer (ET) between protein redox partners. Two-tier energy landscape led to recognize that the 'forward' and 'back' halves of a flash-induced protein-protein electron transfer (ET) photocycle involve different initial configurational ensembles and should respond differently to the modulation of configuration interconversion dynamics.

Experiments into the electrostatic control of electron transfer between myoglobin and cytochrome b5 are presented.