Aqueous-soluble bipyridine cobalt(ii/iii) complexes act as direct redox mediators in photosystem I-based biophotovoltaic devices.

Sustainable energy production is critical for meeting growing worldwide energy demands. Due to its stability and reduction potential, photosystem I (PSI) is attractive as the photosensitizer in biophotovoltaic devices. Herein, we characterize aqueous and organic solvent soluble synthetic bipyridine-based cobalt complexes as redox mediators for PSI-based biophotovoltaics applications. Cobalt-based complexes are not destructive to protein and have appropriate midpoint potentials for electron donation to PSI. We report on PSI stability in organic solvents commonly used in biophotovoltaics. We also show the effects of a mixed organic solvent phase on PSI reduction kinetics, slowing reduction rates approximately 8-38 fold as compared to fully aqueous systems, with implications for dye regeneration rates in PSI-based biophotovoltaics. Further, we show evidence of direct electron transfer from cobalt complexes to PSI. Finally, we report on photocurrent generation from Co mediator-PSI biophotovoltaic devices. Taken together, we discuss the development of novel Co complexes and our ability to fine-tune their characteristics via functional groups and counteranion choice to drive interaction with a biological electron acceptor on multiple levels from redox midpoints, spectral overlap, and solvent requirements, among others. This work suggests that fine-tuning of redox active species for interaction with a biological partner is possible for the creation and improvement of low cost, carbon-neutral energy production in the future.

Electron flow through NDH-1 complexes is the major driver of cyclic electron flow-dependent proton pumping in cyanobacteria.

Cyclic electron flow (CEF) around photosystem I is vital to balancing the photosynthetic energy budget of cyanobacteria and other photosynthetic organisms. The coupling of CEF to proton pumping has long been hypothesized to occur, providing proton motive force (PMF) for the synthesis of ATP with no net cost to [NADPH]. This is thought to occur largely through the activity of NDH-1 complexes, of which cyanobacteria have four with different activities. While a much work has been done to understand the steady-state PMF in both the light and dark, and fluorescent probes have been developed to observe these fluxes in vivo, little has been done to understand the kinetics of these fluxes, particularly with regard to NDH-1 complexes. To monitor the kinetics of proton pumping in Synechocystis sp. PCC 6803, the pH sensitive dye Acridine Orange was used alongside a suite of inhibitors in order to observe light-dependent proton pumping. The assay was demonstrated to measure photosynthetically driven proton pumping and used to measure the rates of proton pumping unimpeded by dark ΔpH. Here, the cyanobacterial NDH-1 complexes are shown to pump a sizable portion of proton flux when CEF-driven and LEF-driven proton pumping rates are observed and compared in mutants lacking some or all NDH-1 complexes. It is also demonstrated that PSII and LEF are responsible for the bulk of light induced proton pumping, though CEF and NDH-1 are capable of generating ~40% of the proton pumping rate when LEF is inactivated.

Enhanced Photocatalytic Hydrogen Production by Hybrid Streptavidin-Diiron Catalysts.

Hybrid protein-organometallic catalysts are being explored for selective catalysis of a number of reactions, because they utilize the complementary strengths of proteins and of organometallic complex. Herein, we present an artificial hydrogenase, StrepH2, built by incorporating a biotinylated [Fe-Fe] hydrogenase organometallic mimic within streptavidin. This strategy takes advantage of the remarkable strength and specificity of biotin-streptavidin recognition, which drives quantitative incorporation of the biotinylated diironhexacarbonyl center into streptavidin, as confirmed by UV/Vis spectroscopy and X-ray crystallography. FTIR spectra of StrepH2 show characteristic peaks at shift values indicative of interactions between the catalyst and the protein scaffold. StrepH2 catalyzes proton reduction to hydrogen in aqueous media during photo- and electrocatalysis. Under photocatalytic conditions, the protein-embedded catalyst shows enhanced efficiency and prolonged activity compared to the isolated catalyst. Transient absorption spectroscopy data suggest a mechanism for the observed increase in activity underpinned by an observed longer lifetime for the catalytic species FeI Fe0 when incorporated within streptavidin compared to the biotinylated catalyst in solution.

Jolly green MOF: confinement and photoactivation of photosystem I in a metal-organic framework.

Photosystem I (PSI) is a ∼1000 kDa transmembrane protein that enables photoactivated charge separation with ∼1 V driving potential and ∼100% quantum efficiency during the photosynthetic process. Although such properties make PSI a potential candidate for integration into bio-hybrid solar energy harvesting devices, the grand challenge in orchestrating such integration rests on rationally designed 3D architectures that can organize and stabilize PSI in the myriad of harsh conditions in which it needs to function. The current study investigates the optical response and photoactive properties of PSI encapsulated in a highly stable nanoporous metal-organic framework (ZIF-8), denoted here as PSI@ZIF-8. The ZIF-8 framework provides a unique scaffold with a robust confining environment for PSI while protecting its precisely coordinated chlorophyll networks from denaturing agents. Significant blue shifts in the fluorescence emissions from UV-vis measurements reveal the successful confinement of PSI in ZIF-8. Pump-probe spectroscopy confirms the photoactivity of the PSI@ZIF-8 composites by revealing the successful internal charge separation and external charge transfer of P700 + and FB - even after exposure to denaturing agents and organic solvents. This work provides greater fundamental understanding of confinement effects on pigment networks, while significantly broadening the potential working environments for PSI-integrated bio-hybrid materials.

Manufactured Brace Modalities for Elbow Stiffness.

Multiple surgical and nonsurgical treatment options exist for patients with elbow stiffness. Many nonsurgical mobilization bracing options have been implemented to increase elbow range of motion. Three of the main bracing options for these patients are turnbuckle, static progressive stretch, and dynamic bracing. The purpose of this study was to review the current literature on turnbuckle, static progressive stretch, and dynamic bracing to provide information for practitioners and patients regarding which brace is more appropriate to use for elbow stiffness. Specifically, the authors compared the protocol and duration of splint use and changes in range of motion outcomes between static progressive and dynamic brace cohorts. A search of PubMed yielded 8 studies meeting inclusion criteria. Overall, although all 3 bracing options are available for patients, these studies found that, based on the evaluated metrics, the static progressive brace was a markedly superior option for patients with elbow stiffness. The time required to wear the static progressive stretch brace was 13 times less than that for the turnbuckle and 5 times less than that for the dynamic devices. Additionally, the high failure rate (10%) and low success rate (29%) of the dynamic brace, compared with the 63% regaining of functional range of motion in the static progressive stretch group, further highlight the benefits of the static progressive stretch brace. [Orthopedics. 2018; 41(1):e127-e135.].

Magnetic Resonance Imaging Predictors of Failure in the Nonoperative Management of Ulnar Collateral Ligament Injuries in Professional Baseball Pitchers.

BACKGROUND: A medial ulnar collateral ligament (UCL) injury of the elbow is an increasingly common injury in professional baseball pitchers. Predictors of success and failure are not well defined for the nonoperative management of these injuries. PURPOSE: To evaluate the efficacy of objective measures to predict failure of the nonoperative management of UCL injuries. STUDY DESIGN: Case-control study; Level of evidence, 3. METHODS: Thirty-two professional pitchers (82%) met inclusion criteria and underwent an initial trial of nonoperative treatment for UCL tears based on clinical and radiological findings. Age, preseason physical examination results, magnetic resonance imaging (MRI) characteristics, and performance metrics were analyzed for these pitchers. Successful nonoperative management was defined as a return to the same level of play or higher for >1 year. Failure was defined as recurrent pain or weakness requiring a surgical intervention after a minimum of 3 months' rest when attempting a return to a throwing rehabilitation program. RESULTS: Thirty-two pitchers (mean age, 22.3 years) who underwent initial nonoperative treatment of UCL injuries were evaluated. Thirty-four percent (11/32) failed and required subsequent ligament reconstruction. Sixty-six percent (21/32) successfully returned to the same level of play for 1 year without a surgical intervention. There was no significant difference seen in physical examination findings or performance metrics between these patients. When comparing MRI findings between the groups, 82% (9/11) ( P < .001) who failed nonoperative management had distal tears, and 81% (17/21) who did not fail had proximal tears ( P < .001). When adjusting for age, location, and evidence of chronic changes on MRI, the likelihood of failing nonoperative management was 12.40 times greater ( P = .020) with a distal tear. No other variable alone or in combination reached significance. When combining the parameters of a high-grade tear and distal location, 88% (7/8) failed nonoperative management. CONCLUSION: In professional pitchers, distal UCL tears showed significantly higher odds of failure with nonoperative management compared with proximal tears. Thus, tear location should be considered when deciding between operative and nonoperative management.

Periarticular Injection After Total Knee Arthroplasty Using Liposomal Bupivacaine vs a Modified Ranawat Suspension: A Prospective, Randomized Study.

BACKGROUND: The purpose of this study is to compare liposomal bupivacaine to a modified (Ranawat) local injection for total knee arthroplasty (TKA). METHODS: This is a prospective, randomized study of 105 consecutive patients undergoing primary TKA. Group A patients received a periarticular injection with liposomal bupivacaine and group B with a mixture of ropivacaine, epinephrine, ketorolac, and clonidine. There were 54 patients in the group A (liposomal bupivacaine) and 51 in group B. RESULTS: There were no differences in the groups with respect to age, sex, and preoperative knee scores. There were no differences with respect to postoperative narcotic usage and knee range of motion. CONCLUSION: Liposomal bupivacaine as a periarticular injection after TKA demonstrated similar pain levels, narcotic usage, and range of motion compared to a modified Ranawat suspension but improved walking distance.

Evolution of reaction center mimics to systems capable of generating solar fuel.

Capturing and converting solar energy via artificial photosynthesis offers an ideal way to limit society's dependence on fossil fuel and its myriad consequences. The development and study of molecular artificial photosynthetic reactions centers and antenna complexes and the combination of these constructs with catalysts to drive the photochemical production of a fuel helps to build the understanding needed for development of future scalable technologies. This review focuses on the study of molecular complexes, design of which is inspired by the components of natural photosynthesis, and covers research from early triad reaction centers developed by the group of Gust, Moore, and Moore to recent photoelectrochemical systems capable of using light to convert water to oxygen and hydrogen.

De novo design of an artificial bis[4Fe-4S] binding protein.

In nature, protein subunits containing multiple iron-sulfur clusters often mediate the delivery of reducing equivalents from metabolic pathways to the active site of redox proteins. The de novo design of redox active proteins should include the engineering of a conduit for the delivery of electrons to and from the active site, in which multiple redox active centers are arranged in a controlled manner. Here, we describe a designed three-helix protein, DSD-bis[4Fe-4S], that coordinates two iron-sulfur clusters within its hydrophobic core. The design exploits the pseudo two-fold symmetry of the protein scaffold, DSD, which is a homodimeric three-helix bundle. Starting from the sequence of the parent peptide, we mutated eight leucine residues per dimer in the hydrophobic core to cysteine to provide the first coordination sphere for cubane-type iron-sulfur clusters. Incorporation of two clusters per dimer is readily achieved by in situ reconstitution and imparts increased stability to thermal denaturation compared to that of the apo form of the peptide as assessed by circular dichroism-monitored thermal denaturation. The presence of [4Fe-4S] clusters in intact proteins is confirmed by UV-vis spectroscopy, gel filtration, analytical ultracentrifugation, and electron paramagnetic resonance spectroscopy. Pulsed electron-electron double-resonance experiments have detected a magnetic dipole interaction between the two clusters ~0.7 MHz, which is consistent with the expected intercluster distance of 29-34 Å. Taken together, our data demonstrate the successful design of an artificial multi-iron-sulfur cluster protein with evidence of cluster-cluster interaction. The design principles implemented here can be extended to the design of multicluster molecular wires.

Catalytic turnover of [FeFe]-hydrogenase based on single-molecule imaging.

Hydrogenases catalyze the interconversion of protons and hydrogen according to the reversible reaction: 2H(+) + 2e(-) ⇆ H(2) while using only the earth-abundant metals nickel and/or iron for catalysis. Due to their high activity for proton reduction and the technological significance of the H(+)/H(2) half reaction, it is important to characterize the catalytic activity of [FeFe]-hydrogenases using both biochemical and electrochemical techniques. Following a detailed electrochemical and photoelectrochemical study of an [FeFe]-hydrogenase from Clostridium acetobutylicum (CaHydA), we now report electrochemical and single-molecule imaging studies carried out on a catalytically active hydrogenase preparation. The enzyme CaHydA, a homologue (70% identity) of the [FeFe]-hydrogenase from Clostridium pasteurianum , CpI, was adsorbed to a negatively charged, self-assembled monolayer (SAM) for investigation by electrochemical scanning tunneling microscopy (EC-STM) techniques and macroscopic electrochemical measurements. The EC-STM imaging revealed uniform surface coverage with sufficient stability to undergo repeated scanning with a STM tip as well as other electrochemical investigations. Cyclic voltammetry yielded a characteristic cathodic hydrogen production signal when the potential was scanned sufficiently negative. The direct observation of the single enzyme distribution on the Au-SAM surface coupled with macroscopic electrochemical measurements obtained from the same electrode allowed the evaluation of a turnover frequency (TOF) as a function of potential for single [FeFe]-hydrogenase molecules.

Solar energy conversion in a photoelectrochemical biofuel cell.

A photoelectrochemical biofuel cell has been developed which incorporates aspects of both an enzymatic biofuel cell and a dye-sensitized solar cell. Photon absorption at a porphyrin-sensitized n-type semiconductor electrode gives rise to a charge-separated state. Electrons and holes are shuttled to appropriate cathodic and anodic catalysts, respectively, allowing the production of electricity, or a reduced fuel, via the photochemical oxidation of a biomass-derived substrate. The operation of this device is reviewed. The use of alternate anodic redox mediators provides insight regarding loss mechanisms in the device. Design strategies for enhanced performance are discussed.

In vitro comparative kinetic analysis of the chloroplast Toc GTPases.

A unique aspect of protein transport into plastids is the coordinate involvement of two GTPases in the translocon of the outer chloroplast membrane (Toc). There are two subfamilies in Arabidopsis, the small GTPases (Toc33 and Toc34) and the large acidic GTPases (Toc90, Toc120, Toc132, and Toc159). In chloroplasts, Toc34 and Toc159 are implicated in precursor binding, yet mechanistic details are poorly understood. How the GTPase cycle is modulated by precursor binding is complex and in need of careful dissection. To this end, we have developed novel in vitro assays to quantitate nucleotide binding and hydrolysis of the Toc GTPases. Here we present the first systematic kinetic characterization of four Toc GTPases (cytosolic domains of atToc33, atToc34, psToc34, and the GTPase domain of atToc159) to permit their direct comparison. We report the KM, Vmax, and Ea values for GTP hydrolysis and the Kd value for nucleotide binding for each protein. We demonstrate that GTP hydrolysis by psToc34 is stimulated by chloroplast transit peptides; however, this activity is not stimulated by homodimerization and is abolished by the R133A mutation. Furthermore, we show peptide stimulation of hydrolytic rates are not because of accelerated nucleotide exchange, indicating that transit peptides function as GTPase-activating proteins and not guanine nucleotide exchange factors in modulating the activity of psToc34. Finally, by using the psToc34 structure, we have developed molecular models for atToc33, atToc34, and atToc159G. By combining these models with the measured enzymatic properties of the Toc GTPases, we provide new insights of how the chloroplast protein import cycle may be regulated.