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Choroidal neovascular membrane (CNVM) formation is a well-documented sight-threatening complication of posterior segment intraocular inflammation (PSII). The aim of this article is to review the basic and clinical science literature on the pathogenesis of CNVM formation in PSII and to present results of a case series. We searched the literature using the mesh terms- inflammation, CNVM, age-related macular degeneration, immunosuppression, photodynamic therapy, steroids, vascular endothelial growth factors and posterior uveitis. Additionally, we evaluated the visual outcome of and clinical response to our standard treatment protocol involving a combination treatment for young patients with inflammatory CNVM. The development of CNVM in PSII is promulgated by infiltrating myeloid cells as well as choroidal and retinal myeloid cell activation, subsequent vascular endothelial growth factors, cytokine and chemokine production and complement activation acting in consort to mediate angiogenic responses. No clear standard of care currently exists for the treatment of inflammatory CNVM and various combinations have been tried. Using our combination treatment, visual acuity improved in four, stabilized in one and worsened in four patients. Though significant advances have occurred in the understanding of the pathogenesis and management of this condition, optimizing therapeutic regimens will require further well-constructed prospective cohort series. [ABSTRACT FROM AUTHOR]

Specialized or secondary metabolites are small molecules of biological origin, often showing potent biological activities with applications in agriculture, engineering and medicine. Usually, the biosynthesis of these natural products is governed by sets of co-regulated and physically clustered genes known as biosynthetic gene clusters (BGCs). To share information about BGCs in a standardized and machine-readable way, the Minimum Information about a Biosynthetic Gene cluster (MIBiG) data standard and repository was initiated in 2015. Since its conception, MIBiG has been regularly updated to expand data coverage and remain up to date with innovations in natural product research. Here, we describe MIBiG version 4.0, an extensive update to the data repository and the underlying data standard. In a massive community annotation effort, 267 contributors performed 8304 edits, creating 557 new entries and modifying 590 existing entries, resulting in a new total of 3059 curated entries in MIBiG. Particular attention was paid to ensuring high data quality, with automated data validation using a newly developed custom submission portal prototype, paired with a novel peer-reviewing model. MIBiG 4.0 also takes steps towards a rolling release model and a broader involvement of the scientific community. MIBiG 4.0 is accessible online at https://mibig.secondarymetabolites.org/., (© The Author(s) 2024. Published by Oxford University Press on behalf of Nucleic Acids Research.)

Cell-free protein synthesis (CFPS) has emerged as an attractive platform for biotechnology and synthetic biology due to its numerous advantages to cell-based technologies for specific applications. CFPS can be faster, less sensitive to metabolite toxicity, and amenable to systems that are not easily genetically manipulated. Due to these advantages, a promising application of CFPS is to characterize biosynthetic gene clusters, particularly those harbored within the genomes of microorganisms that generate secondary metabolites, otherwise known as natural products. In the postgenomic era, genome sequencing has revealed an incredible wealth of metabolic diversity. However, far more of these pathways are termed "cryptic," i.e., unable to be produced under standard laboratory conditions than have been characterized. A major barrier to characterizing these cryptic natural products using CFPS is that many of these pathways require utilization of complex cofactors, many of which to date are not recycled efficiently or in an economically viable fashion. In this perspective, we outline strategies to regenerate cofactors relevant to secondary metabolite production in CFPS. This includes adenosine 5'-triphosphate, coenzyme A, redox cofactors (iron-sulfur clusters, nicotinamide adenine dinucleotide phosphate, flavin adenine dinucleotide), all of which play a crucial role in important biosynthetic enzymes. Such advances in cofactor recycling enable continuous production of complex metabolites in CFPS and expand the utility of this emergent platform., Competing Interests: Competing interestsThe authors declare no competing interests., (© The Author(s) 2024.)

Some of the most metabolically diverse species of bacteria (e.g., Actinobacteria) have higher GC content in their DNA, differ substantially in codon usage, and have distinct protein folding environments compared to tractable expression hosts like Escherichia coli. Consequentially, expressing biosynthetic gene clusters (BGCs) from these bacteria in E. coli often results in a myriad of unpredictable issues with regard to protein expression and folding, delaying the biochemical characterization of new natural products. Current strategies to achieve soluble, active expression of these enzymes in tractable hosts can be a lengthy trial-and-error process. Cell-free expression (CFE) has emerged as a valuable expression platform as a testbed for rapid prototyping expression parameters. Here, we use a type III polyketide synthase from Streptomyces griseus, RppA, which catalyzes the formation of the red pigment flaviolin, as a reporter to investigate BGC refactoring techniques. We applied a library of constructs with different combinations of promoters and rppA coding sequences to investigate the synergies between promoter and codon usage. Subsequently, we assess the utility of cell-free systems for prototyping these refactoring tactics prior to their implementation in cells. Overall, codon harmonization improves natural product synthesis more than traditional codon optimization across cell-free and cellular environments. More importantly, the choice of coding sequences and promoters impact protein expression synergistically, which should be considered for future efforts to use CFE for high-yield protein expression. The promoter strategy when applied to RppA was not completely correlated with that observed with GFP, indicating that different promoter strategies should be applied for different proteins. In vivo experiments suggest that there is correlation, but not complete alignment between expressing in cell free and in vivo. Refactoring promoters and/or coding sequences via CFE can be a valuable strategy to rapidly screen for catalytically functional production of enzymes from BCGs, which advances CFE as a tool for natural product research., (© 2024. The Author(s).)

Prevention services can promote public health by building protective factors and reducing maltreatment risk. Yet, engaging caregivers in prevention services presents a unique set of challenges. Measurement studies are important first steps to increase the knowledge of caregiver engagement in prevention services. The purpose of this scoping review was to investigate how family engagement has been measured and operationalized in the studies of maltreatment prevention/positive parenting programs. The review examined quantitative and mixed methods studies conducted in the U.S., which measured multiple dimensions of client engagement, including behavioral, attitudinal, and relational domains. A total of 88 studies selected from PubMed, CINAHL, ERIC, PsycINFO, Social Work Abstracts, Academic Search Premier, and Web of Science were included in this review. Results indicated that studies examine engagement constructs in all three domains of engagement with a primary focus on behavioral engagement. The attitudinal and relational engagement was mostly assessed through general satisfaction surveys, and a limited number of studies utilized validated measures to assess those constructs. While most studies reported acceptable internal reliabilities, only two studies reported other dimensions of psychometric qualities. Only one validated measure was found, which assessed client perceptions of provider cultural competence. More measurement studies are needed to further incorporate multiple dimensions of engagement into the studies of maltreatment prevention programs, which can inform the effort to develop tailored implementation strategies to fully engage various groups of parents in maltreatment prevention programs., Competing Interests: Declaration of Conflicting InterestsThe authors declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.

High GC bacteria from the genus Streptomyces harbor expansive secondary metabolism. The expression of biosynthetic proteins and the characterization and identification of biological "parts" for synthetic biology purposes from such pathways are of interest. However, the high GC content of proteins from actinomycetes in addition to the large size and multi-domain architecture of many biosynthetic proteins (such as non-ribosomal peptide synthetases; NRPSs, and polyketide synthases; PKSs often called "megasynthases") often presents issues with full-length translation and folding. Here we evaluate a non-ribosomal peptide synthetase (NRPS) from Streptomyces lavenduale, a multidomain "megasynthase" gene that comes from a high GC (72.5%) genome. While a preliminary step in revealing differences, to our knowledge this presents the first head-to-head comparison of codon-optimized sequences versus a native sequence of proteins of streptomycete origin heterologously expressed in E. coli. We found that any disruption in co-translational folding from codon mismatch that reduces the titer of indigoidine is explainable via the formation of more inclusion bodies as opposed to compromising folding or posttranslational modification in the soluble fraction. This result supports that one could apply any refactoring strategies that improve soluble expression in E. coli without concern that the protein that reaches the soluble fraction is differentially folded., (Copyright © 2023 Elsevier Inc. All rights reserved.)

In primary metabolism, fatty acid synthases (FASs) biosynthesize fatty acids via sequential Claisen-like condensations of malonyl-CoA followed by reductive processing. Likewise, polyketide synthases (PKSs) share biosynthetic logic with FAS which includes utilizing the same precursors and cofactors. However, PKS biosynthesize structurally diverse, complex secondary metabolites, many of which are pharmaceutically relevant. This digest covers examples of interconnected biosynthesis between primary and secondary metabolism in fatty acid and polyketide metabolism. Taken together, further understanding the biosynthetic linkage between polyketide biosynthesis and fatty acid biosynthesis may lead to improved discovery and production of novel drug leads from polyketide metabolites., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

Lysate-based cell-free expression (CFE) systems are accessible platforms for expressing proteins that are difficult to synthesize in vivo , such as nonribosomal peptide synthetases (NRPSs). NRPSs are large (>100 kDa), modular enzyme complexes that synthesize bioactive peptide natural products. This synthetic process is analogous to transcription/translation (TX/TL) in lysates, resulting in potential resource competition between NRPS expression and NRPS activity in cell-free environments. Moreover, CFE conditions depend on the size and structure of the protein. Here, a reporter system for rapidly investigating and optimizing reaction environments for NRPS CFE is described. This strategy is demonstrated in E. coli lysate reactions using blue pigment synthetase A (BpsA), a model NRPS, carrying a C-terminal tetracysteine (TC) tag which forms a fluorescent complex with the biarsenical dye, FlAsH. A colorimetric assay was adapted for lysate reactions to detect the blue pigment product, indigoidine, of cell-free expressed BpsA-TC, confirming that the tagged enzyme is catalytically active. An optimized protocol for end point TC/FlAsH complex measurements in reactions enables quick comparisons of full-length BpsA-TC expressed under different reaction conditions, defining unique requirements for NRPS expression that are related to the protein's catalytic activity and size. Importantly, these protein-dependent CFE conditions enable higher indigoidine titer and improve the expression of other monomodular NRPSs. Notably, these conditions differ from those used for the expression of superfolder GFP (sfGFP), a common reporter for optimizing lysate-based CFE systems, indicating the necessity for tailored reporters to optimize expression for specific enzyme classes. The reporter system is anticipated to advance lysate-based CFE systems for complex enzyme synthesis, enabling natural product discovery.

Child maltreatment is a highly prevalent public health concern that contributes to morbidity and mortality in childhood and short- and long-term health consequences that persist into adulthood. Past research suggests that social determinants of health such as socioeconomic status and intergenerational trauma are highly correlated with child maltreatment. With support from the U.S. Children's Bureau, the Ohio Children's Trust Fund is currently piloting the Family Success Network, a primary child maltreatment prevention strategy in Northeast Ohio that seeks to address these social determinants through pillars of service that include family coaching, financial assistance, financial education, parenting education, and basic life skills training. This study highlights the initial development phase of a pilot study. Plans for in-depth process and outcome evaluations are discussed. The project seeks to improve family functioning and reduce child protective services involvement and foster care entry in an economically disadvantaged region.

Bacterial modular type I polyketide synthases (PKSs) are complex multidomain assembly line proteins that produce a range of pharmaceutically relevant molecules with a high degree of stereochemical control. Due to their colinear properties, they have been considerable targets for rational biosynthetic pathway engineering. Among the domains harbored within these complex assembly lines, ketoreductase (KR) domains have been extensively studied with the goal of altering their stereoselectivity by site-directed mutagenesis, as they confer much of the stereochemical complexity present in pharmaceutically active reduced polyketide scaffolds. Here we review all efforts to date to perform site-directed mutagenesis on PKS KRs, most of which have been done in the context of excised KR domains on model diffusible substrates such as β-keto N-acetyl cysteamine thioesters. We also discuss the challenges around translating the findings of these studies to alter stereocontrol in the context of a complex multidomain enzymatic assembly line., (© 2020 Wiley-VCH GmbH.)

Polyketide synthase (PKS) engineering is an attractive method to generate new molecules such as commodity, fine and specialty chemicals. A significant challenge is re-engineering a partially reductive PKS module to produce a saturated β-carbon through a reductive loop (RL) exchange. In this work, we sought to establish that chemoinformatics, a field traditionally used in drug discovery, offers a viable strategy for RL exchanges. We first introduced a set of donor RLs of diverse genetic origin and chemical substrates  into the first extension module of the lipomycin PKS (LipPKS1). Product titers of these engineered unimodular PKSs correlated with chemical structure similarity between the substrate of the donor RLs and recipient LipPKS1, reaching a titer of 165 mg/L of short-chain fatty acids produced by the host  Streptomyces albus J1074. Expanding this method to larger intermediates that require bimodular communication, we introduced RLs of divergent chemosimilarity into LipPKS2 and determined triketide lactone production. Collectively, we observed a statistically significant correlation between atom pair chemosimilarity and production, establishing a new chemoinformatic method that may aid in the engineering of PKSs to produce desired, unnatural products.

Modular polyketide synthases (PKSs) are a multidomain megasynthase class of biosynthetic enzymes that have great promise for the development of new compounds, from new pharmaceuticals to high value commodity and specialty chemicals. Their colinear biosynthetic logic has been viewed as a promising platform for synthetic biology for decades. Due to this colinearity, domain swapping has long been used as a strategy to introduce molecular diversity. However, domain swapping often fails because it perturbs critical protein-protein interactions within the PKS. With our increased level of structural elucidation of PKSs, using judicious targeted mutations of individual residues is a more precise way to introduce molecular diversity with less potential for global disruption of the protein architecture. Here we review examples of targeted point mutagenesis to one or a few residues harbored within the PKS that alter domain specificity or selectivity, affect protein stability and interdomain communication, and promote more complex catalytic reactivity., (© 2020 Production and hosting by Elsevier B.V. on behalf of KeAi Communications Co., Ltd.)

The conversion of biomass to biofuels presents a solution to one of the largest global challenges of our era, climate change. A critical part of this pipeline is the process of breaking down cellulosic sugars from plant matter to be used by microbes containing biosynthetic pathways that produce biofuels or bioproducts. In this inquiry-based course, students complete a research project that isolates cellulase-producing bacteria from samples collected from the environment. After obtaining isolates, the students characterize the production of cellulases. Students then amplify and sequence the 16S rRNA genes of confirmed cellulase producers and use bioinformatic methods to identify the bacterial isolates. Throughout the course, students learn about the process of generating biofuels and bioproducts through the deconstruction of cellulosic biomass to form monosaccharides from the biopolymers in plant matter. The program relies heavily on active learning and enables students to connect microbiology with issues of sustainability. In addition, it provides exposure to basic microbiology, molecular biology, and biotechnology laboratory techniques and concepts. The described activity was initially developed for the Introductory College Level Experience in Microbiology (iCLEM) program, a research-based immersive laboratory course at the US Department of Energy Joint BioEnergy Institute. Originally designed as an accelerated program for high-potential, low-income, high school students (11th-12th grade), this curriculum could also be implemented for undergraduate coursework in a research-intensive laboratory course at a two- or four-year college or university.

Background and Objectives: There is a growing number of adults with long-term mobility impairment aging into the older adult population. Little is known about the experiences of these individuals in maintaining activities of daily living (ADLs) and instrumental activities of daily living (IADLs) as they face age-related changes in addition to a pre-existing mobility impairment., Research Design and Methods: Through in-home interviews with 21 participants (ages 52-86) with long-term mobility impairment, the present study employed a qualitative description design to explore perceptions of how and why select ADL/IADL routines (e.g., bed transfer, toileting) have changed over time. The selection, optimization, and compensation (SOC) model was used as a framework to organize participants' adaptations., Results: Among the ADL/IADL routine changes mentioned, elective selection strategies, in which a person continues to work at maintaining a task, were more frequently endorsed than loss-based selection strategies, in which a person does a task less or gets help from someone. Findings suggest that this population is actively adapting their routines to preserve their involvement in, and frequency of doing, these ADLs/IADLs. Counter to expectation, perceived age-related changes underlying activity routine changes were subtle and generally did not include sensory and cognitive declines., Discussion and Implications: Findings provide insights into the difficulties adults with long-term mobility impairment experience as they age, as well as the adaptations they employ to overcome those challenges. Results highlight the need for customizable, mobility supports (e.g., assistive technologies, home modifications) that can adjust to an individual's changing abilities across the life span., (© The Author 2017. Published by Oxford University Press on behalf of The Gerontological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

pSC101 is a narrow host range, low-copy plasmid commonly used for genetically manipulating Escherichia coli. As a byproduct of a genetic screen for a more sensitive lactam biosensor, we identified multiple novel mutations that increase the copy number of plasmids with the pSC101 origin. All mutations identified in this study occurred on plasmids which also contained at least one mutation localized to the RepA protein encoded within the origin. Homology modelling predicts that many of these mutations occur within the dimerization interface of RepA. Mutant RepA resulted in plasmid copy numbers between ~31 and ~113 copies/cell, relative to ~5 copies/cell in wild-type pSC101 plasmids. Combining the mutations that were predicted to disrupt multiple contacts on the dimerization interface resulted in copy numbers of ~500 copies/cell, while also attenuating growth in host strains. Fluorescent protein production expressed from an arabinose-inducible promoter on mutant origin derived plasmids did correlate with copy number. Plasmids harboring RepA with one of two mutations, E83K and N99D, resulted in fluorescent protein production similar to that from p15a- (~20 copies/cell) and ColE1- (~31 copies/cell) based plasmids, respectively. The mutant copy number variants retained compatibility with p15a, pBBR, and ColE1 origins of replication. These pSC101 variants may be useful in future metabolic engineering efforts that require medium or high-copy vectors compatible with p15a- and ColE1-based plasmids.

ClusterCAD is a web-based toolkit designed to leverage the collinear structure and deterministic logic of type I modular polyketide synthases (PKSs) for synthetic biology applications. The unique organization of these megasynthases, combined with the diversity of their catalytic domain building blocks, has fueled an interest in harnessing the biosynthetic potential of PKSs for the microbial production of both novel natural product analogs and industrially relevant small molecules. However, a limited theoretical understanding of the determinants of PKS fold and function poses a substantial barrier to the design of active variants, and identifying strategies to reliably construct functional PKS chimeras remains an active area of research. In this work, we formalize a paradigm for the design of PKS chimeras and introduce ClusterCAD as a computational platform to streamline and simplify the process of designing experiments to test strategies for engineering PKS variants. ClusterCAD provides chemical structures with stereochemistry for the intermediates generated by each PKS module, as well as sequence- and structure-based search tools that allow users to identify modules based either on amino acid sequence or on the chemical structure of the cognate polyketide intermediate. ClusterCAD can be accessed at https://clustercad.jbei.org and at http://clustercad.igb.uci.edu., (© The Author(s) 2017. Published by Oxford University Press on behalf of Nucleic Acids Research.)

Given the high prevalence of ankle fractures and morbidity of malalignment after fixation, an appropriate anatomic relationship between the distal fibula and adjacent tibia and talus is important. The tip of the lateral malleolus of the fibula has often been described to be at the level of the lateral talar process. However, no studies to date have examined the relationship of the distal fibular tip to the lateral process of the talus. We assessed 66 weightbearing mortise radiographs for variability of the distal fibular tip in relation to the lateral process of the talus. The subjects were all skeletally mature, with a mean age of 45.3 ± 14.6 years. We used a paired t test with a null hypothesis that the true mean difference in the distance from the distal fibula to the lateral process was equal to 0. The mean distance of the distal tip of the fibula was 0.257 ± 0.127 cm proximal to the tip of the lateral process of the talus. The 95% confidence interval was 0.226 to 0.288. Of the 66 subjects, 65 had the distal tip of the fibula proximal to the lateral process of the talus, corresponding to a negative fibular variance. In the remaining subject, the distal tip of the fibula was at the same level of the tip as the lateral process of the talus. The distal tip of the fibula is most commonly not at the level of the talus lateral process, as often described in published reports. Instead, it has a variance analogous to the relationship between the lengths of the ulna compared with the radius. The distal tip of the fibula in our study was more often proximal to the tip of the lateral process of the talus and can be described as a negative fibular variance, or "fibula minus.", (Copyright © 2017 The American College of Foot and Ankle Surgeons. Published by Elsevier Inc. All rights reserved.)

Streptomyces genomes have a high G + C content and typically use an ATG or GTG codon to initiate protein synthesis. Although gene-finding tools perform well in low GC genomes, it is known that the accuracy in predicting a translational start site (TSS) is much less for high GC genomes. LipPks1 is a Streptomyces-derived, well-characterized modular polyketide synthase (PKS). Using this enzyme as a model, we experimentally investigated the effects of alternative TSSs using a heterologous host, Streptomyces venezuelae. One of the TSSs employed boosted the protein level by 59-fold and the product yield by 23-fold compared to the originally annotated start codon. Interestingly, a structural model of the PKS indicated the presence of a structural motif in the N-terminus, which may explain the observed different protein levels together with a proline and arginine-rich sequence that may inhibit translational initiation. This structure was also found in six other modular PKSs that utilize noncarboxylated starter substrates, which may guide the selection of optimal TSSs in conjunction with start-codon prediction software.

Metabolic engineering efforts toward rewiring metabolism of cells to produce new compounds often require the utilization of non-native enzymatic machinery that is capable of producing a broad range of chemical functionalities. Polyketides encompass one of the largest classes of chemically diverse natural products. With thousands of known polyketides, modular polyketide synthases (PKSs) share a particularly attractive biosynthetic logic for generating chemical diversity. The engineering of modular PKSs could open access to the deliberate production of both existing and novel compounds. In this review, we discuss PKS engineering efforts applied at both the protein and cellular level for the generation of a diverse range of chemical structures, and we examine future applications of PKSs in the production of medicines, fuels and other industrially relevant chemicals.

In an effort to uncover the structural motifs and biosynthetic logic of the relatively uncharacterized trans-acyltransferase polyketide synthases, we have begun the dissection of the enigmatic dehydrating bimodules common in these enzymatic assembly lines. We report the 1.98 Å resolution structure of a ketoreductase (KR) from the first half of a type A dehydrating bimodule and the 2.22 Å resolution structure of a dehydratase (DH) from the second half of a type B dehydrating bimodule. The KR, from the third module of the bacillaene synthase, and the DH, from the tenth module of the difficidin synthase, possess features not observed in structurally characterized homologs. The DH architecture provides clues for how it catalyzes a unique double dehydration. Correlations between the chemistries proposed for dehydrating bimodules and bioinformatic analysis indicate that type A dehydrating bimodules generally produce an α/β-cis alkene moiety, while type B dehydrating bimodules generally produce an α/β-trans, γ/δ-cis diene moiety., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Advances in retooling microorganisms have enabled bioproduction of 'drop-in' biofuels, fuels that are compatible with existing spark-ignition, compression-ignition, and gas-turbine engines. As the majority of petroleum consumption in the United States consists of gasoline (47%), diesel fuel and heating oil (21%), and jet fuel (8%), 'drop-in' biofuels that replace these petrochemical sources are particularly attractive. In this review, we discuss the application of aldehyde decarbonylases to produce gasoline substitutes from fatty acid products, a recently crystallized reductase that could hydrogenate jet fuel precursors from terpene synthases, and the exquisite control of polyketide synthases to produce biofuels with desired physical properties (e.g., lower freezing points). With our increased understanding of biosynthetic logic of metabolic pathways, we discuss the unique advantages of fatty acid, terpene, and polyketide synthases for the production of bio-based gasoline, diesel and jet fuel., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Objective: To assess the usability and acceptance of activity tracking technologies by older adults., Method: First in our multimethod approach, we conducted heuristic evaluations of two activity trackers that revealed potential usability barriers to acceptance. Next, questionnaires and interviews were administered to 16 older adults ( M age = 70, SD age = 3.09, range age = 65-75) before and after a 28-day field study to understand facilitators and additional barriers to acceptance. These measurements were supplemented with diary and usage data and assessed whether and why users overcame usability issues., Results: The heuristic evaluation revealed usability barriers in System Status Visibility, Error Prevention, and Consistency and Standards. The field study revealed additional barriers (e.g., accuracy, format) and acceptance-facilitators (e.g., goal tracking, usefulness, encouragement)., Discussion: The acceptance of wellness management technologies, such as activity trackers, may be increased by addressing acceptance-barriers during deployment (e.g., providing tutorials on features that were challenging, communicating usefulness).

Modular polyketide synthase ketoreductases can set two chiral centers through a single reduction. To probe the basis of stereocontrol, a structure-activity relationship study was performed with three α-methyl, β-ketothioester substrates and four ketoreductases. Since interactions with the β-ketoacyl moiety were found to be most critical, residues implicated in contacting this moiety were mutated. Two mutations were sufficient to completely reverse the stereoselectivity of the model ketoreductase EryKR1, converting it from an enzyme that generates (2S,3R)-products into one that yields (2S,3S)-products.

This study examined potential issues faced by older adults in managing their homes and their proposed solutions for overcoming hypothetical difficulties. Forty-four diverse, independently living older adults (66-85) participated in structured group interviews in which they discussed potential solutions to manage difficulties presented in four scenarios: perceptual, mobility, physical, and cognitive difficulties. The proposed solutions were classified using the Selection, Optimization, and Compensation (SOC) model. Participants indicated they would continue performing most tasks and reported a range of strategies to manage home maintenance challenges. Most participants reported that they would manage home maintenance challenges using compensation; the most frequently mentioned compensation strategy was using tools and technologies. There were also differences across the scenarios: Optimization was discussed most frequently with perceptual and cognitive difficulty scenarios. These results provide insights into supporting older adults' potential needs for aging-in-place and provide evidence of the value of the SOC model in applied research., (© The Author(s) 2012.)

Individuals are becoming more involved in managing their own health. Health self-management technologies have the potential to help older adults remain well by promoting exercise and a good diet. However, older adults may or may not decide to adopt wellness management technologies. Adoption is a process and the intent to adopt may change over time. Sixteen older adults (8 females; M age =70.06, SD =3.09; range=65-75) used one of two wellness management technologies (the Fitbit One or myfitnesspal.com) over a 28-day period. Initially, all participants were open or neutral to adopting their technologies. After 28 days, 12 participants intended to adopt and 4 participants did not intend to adopt. The diary data revealed that over time, adopters made more positive comments than non-adopters. Both adopters and non-adopters mentioned perceived ease of use praises and complaints, whereas only adopters mentioned praises regarding usefulness. Results are interpreted within the frameworks of the unified theory of acceptance and use of technology (Venkatesh et al., 2003) and the diffusion of innovation (Rogers, 2003). Changes in intent to adopt suggest that experience is important in the adoption decision. Adoption of wellness management technologies by older adults may increase if designers attend to the perceived ease of use and perceived usefulness factors identified in this study.

Mechanically sensitive biocomposites comprised of fluorescent proteins report stress through distinct pathways. Whereas a composite containing an enhanced yellow fluorescent protein (eYFP) exhibited hypsochromic shifts in its fluorescence emission maxima following compression, a composite containing a modified green fluorescent protein (GFPuv) exhibited fluorescence quenching under the action of mechanical force. These ratio- and intensiometric sensors demonstrate that insights garnered from disparate fields (that is, polymer mechanochemistry and biophysics) can be harnessed to guide the rational design of new classes of biomechanophore-containing materials., (© 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Self-management of health is becoming increasingly important in today's healthcare climate. Activity monitoring technologies have the potential to support health self-management by tracking, storing, compiling, and providing feedback about an individual's engagement in movement activities. Older adults represent a fast growing segment of the population who may benefit from such technologies. To understand how to facilitate technology acceptance and adoption, more information is needed about older adults' attitudes and usage of such technologies. Eight older adult participants ( M age = 65.0 years; SD = 3.2; range = 61-69) used one of four activity monitoring technologies in their own homes for two weeks. Attitudes and usability issues were assessed and evaluated within a technology acceptance framework. Participants' initial attitudes were positive, but after using the technology for two weeks, attitudes were mixed. Three participants indicated they would continue using the technology, whereas five said they would abandon the technology. These data offer insight into older adults' use of and attitudes toward activity monitoring technologies and provide improvement opportunities for designers. The results suggest that efforts should focus on conveying the usefulness and personal benefits of activity monitoring technologies specific to older adults.

A fundamental aspect of successful illness self-management for people with serious mental illnesses is the ability to advocate for themselves in health and rehabilitation settings. This study reports findings from a randomized controlled trial comparing propensity for patient self-advocacy among those who received a peer-led mental illness self-management intervention called Wellness Recovery Action Planning (WRAP) and those who received usual care. Outcomes were self-reported engagement in self-advocacy with service providers, and the relationship between patient self-advocacy and other key recovery outcomes. In a multivariable analysis, at immediate post-intervention and 6-month follow-up, WRAP participants were significantly more likely than controls to report engaging in self-advocacy with their service providers. Higher self-advocacy also was associated with greater hopefulness, better environmental quality of life, and fewer psychiatric symptoms among the intervention group. These findings provide additional support for the positive impact of peer-led illness self-management on mental health recovery.

PKS biocatalysis: The terminal module of erythromycin synthase was used for the in vitro production of chiral triketide lactones. Combining cofactor regeneration, substrate truncation, and enzymatic promiscuity afforded a scalable strategy to generate these molecules from abundant racemic and achiral precursors. The described biocatalytic platform thus facilitates the application and study of enzymes within PKS modules., (Copyright © 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Objective: The purpose of this study was to test the effectiveness of the Wellness Recovery Action Planning (WRAP) self-management intervention in reducing depression and anxiety and in increasing self-perceived recovery among individuals with a serious mental illness., Methods: Participants were recruited from outpatient community mental health settings in six Ohio communities: Canton, Cleveland, Columbus, Dayton, Lorain, and Toledo. With a single-blind, randomized controlled trial design, 519 individuals were assigned to WRAP or to services as usual and assessed at baseline and at two- and eight-month follow-ups. The intervention consisted of eight weekly 2.5-hour sessions delivered by peers in recovery from serious mental illness who were certified WRAP educators., Results: The mean number of WRAP sessions attended was five, and fidelity ranged from 90% to 92%. Analysis using mixed-effects random regression revealed interactions of study condition by time in each outcome area. Compared with the control group, intervention participants reported significantly greater reduction over time in Brief Symptom Inventory depression and anxiety subscales and significantly greater improvement in total Recovery Assessment Scale scores as well as the subscales measuring personal confidence and goal orientation., Conclusions: Training in mental illness self-management reduced depression and anxiety and improved participants' self-perceived recovery over time. Results confirmed the importance of WRAP as part of a group of evidence-based, recovery-oriented interventions.

Age-related declines in capabilities may compromise older adults' ability to maintain their homes thus threatening successful aging in place. Structured interviews were conducted with forty-four independently living older adults (M(age) = 76.1, SD = 4.7) to discuss difficult home maintenance tasks and how they managed those tasks. Solutions to managing difficulties were categorized as person-related or environment-related. The majority (85%) of responses were person-related solutions. An understanding of the specific challenges that older adults face in maintaining their homes can guide redesign efforts and interventions to effectively support older adults' desire to age in place.

Older adults (n = 113) participated in focus groups discussing their use of and attitudes about technology in the context of their home, work, and healthcare. Participants reported using a wide variety of technology items, particularly in their homes. Positive attitudes (i.e., likes) outnumbered negative attitudes (i.e., dislikes), suggesting that older adults perceive the benefits of technology use to outweigh the costs of such use. Positive attitudes were most frequently related to how the technology supported activities, enhanced convenience, and contained useful features. Negative attitudes were most frequently associated with technology creating inconveniences, unhelpful features, as well as security and reliability concerns. Given that older adults reported more positive than negative attitudes about the technologies they use, these results contradict stereotypes that older adults are afraid or unwilling to use technology. These findings also highlight the importance of perceived benefits of use and ease of use for models of technology acceptance. Emphasizing the benefits of technology in education and training programs may increase future technology adoption.

A goal of many older adults is to remain in their own homes as they age (Beyond 50.05 Survey, 2005). However, a detailed assessment is lacking of the needs of older adults as they age in place. Using focus groups, twenty-six independently living older adults (mean age 78.8 years) from the Atlanta metropolitan area were asked to describe the tasks they perform to maintain their homes, as well as any difficulties they have performing these tasks. Participants described performing a wide range of tasks and focused primarily on physical difficulties. However, participants also reported solutions to manage these difficulties that fell into three broad categories: "Cessation," "Perseverance," and "Compensation." These categories represent classes of opportunities for interventions that may help older adults remain independent in their homes longer. By understanding the nature of home maintenance problems older adults encounter while aging in place, interventions and redesign efforts can be more effective. These data suggest that interventions should start with answering physical issues.

Visual line graphs are a prevalent form of communication as they provide a pictorial means to display relationships between entities. As such, understanding the cognitive resources required in processing line graphs would inform designers how to optimize the use of graphical displays. This study systematically investigated how graph task performance changes as a function of attention allocation (full or divided) and concurrent memory task (spatial or verbal). Twenty-four younger adults (mean age 19.2 years) completed either a trend comparison task or a point estimation task and a concurrent spatial or verbal task. Trend comparison performance did not significantly differ between the full and divided attention conditions; mean performance for all conditions was over 90% accurate. Interestingly, participants' point estimation performance was significantly better for the two divided attention conditions compared to the full attention condition which may be attributed to a motivational or stimulus effect. This study provides a base from which more research can be conducted to understand the verbal and spatial resources required in graph comprehension.

Older adults may benefit from using technology in their everyday lives. However, adults over 65 may need more training than their younger counterparts given they have had less experience with technology. In this study, 113 adults between the ages of 65 and 85 participated in focus group interviews discussing their training needs and preferences for technology items used in the home. Participants expressed an interest in receiving additional training, particularly for specific tasks. Participants also discussed preferences for various characteristics of training, such as who should conduct the training and for their preferred method of training. One of the most frequently discussed preferences was for self-training using text materials, such as a manual.

Descriptively many Protestant perspectives on access to health care share much in common with secular accounts ... perhaps too much. A normative account of a Protestant perspective on access to health care must be perceptively qualified by the Christ story. A genuinely theocratic approach may not lead one to the conclusion that the faith commits us to the notion of equality; rather the faith commits us to the notion of care. What counts as care may not be the same in every instance (if this is indeed what equality means; in ordinary use, equality is tantamount to sameness). The Christ story reminds us who we are, beloved, sinful, redeemed. Acting out this story is to accept, to give, and to receive reconciliation. For many Christians this will mean that we are called to deny ourselves "equal" access to the system (as if equality actually existed!).

Twelve Holstein-Friesian calves were divided into two groups, one of which was infected with Eimeria zuernii. Fecal oocyst output, weight changes and various blood, cellular, protein and biochemical constituents were examined for both groups. Maximal fecal oocyst output occurred 21 days after infection. Both groups of calves gained weight in a linear fashion until day 21 postinfection when the infected group lost weight rapidly. The packed cell volume of the infected calves appeared markedly reduced. Although there was a reduction in plasma proteins it did not appear significant. There was a significant reduction in plasma Na+ and Cl- ions of the infected calves. There were no significant changes in the other blood constituents examined.

Association of alkaline phosphatase with silicic acid in precipitates formed in dilute solution was studied as a model for the nonspecific reaction between silicic acid and protein. Precipitates contained 68-83% of the silicic acid and 52-83% of the enzyme in the original mixture and were in the form of aggregates of roundish particles 150-800 nm in diameter. Enzyme protein formed a tightly bound layer on the surface of particles formed in solutions of freshly prepared silicic acid. The similarity between the ultrastructural features of precipitates from solutions of silicic acid and of internal portions of siliceous urinary calculi from cattle suggests that deposition of silica during development of such calculi is due, at least in part, to the interaction of protein with silicic acid in urine.

Blocks of tissue were removed from various locations in the bovine digestive tract and fixed and processed for transmission and scanning electron microscopy by techniques that retained adherent bacteria. The distribution of bacteria on the surface of epithelial cells was examined by scanning electron microscopy. This showed intermittent colonization of the epithelia with the formation of occasional microcolonies of morphologically similar bacterial cells. Transmission electron microscopy of ruthenium red-stained material showed the presence of both the glycocalyx of the bovine epithelial cells and fibrous carbohydrate coats surrounding adherent bacteria. The carbohydrate coats appeared to mediate the attachment of bacteria to the epithelium, to food particles, and to each other so that microcolonies were formed. Careful examination of the bacterial colonization of keratinized cells in the process of being sloughed from the surface of the stratified squamous epithelium of the rumen showed that these dead cells were digested by adherent bacteria of a limited number of morphological types. The spatial relationship of this mixed, adherent, microbial population to living and dead epithelial cells and to food particles indicates that digestive processes of some importance may be accomplished by this stationary component of the microbial flora of the digestive tract.

Prospective study of patients admitted to a hospital for closed head injury showed that nine patients (nearly 3%) became mute for varying periods despite recovery of consciousness and communication through a nonspeech channel. Computed tomography (CT) showed subcortical lesions situated primarily in the putamen and internal capsule of four patients, whereas four of the five patients without subcortical lesions had left-hemisphere cortical injury. The patients without subcortical injury visualized by CT exhibited a longer duration of impaired consciousness consistent with severe diffuse brain injury and they showed more long-term linguistic deficits. We related our findings to recent studies of atypical aphasia after occlusive vascular lesions of the basal ganglia.