Assistive Technology and Older Adults: Education to Support Evidence-Based Practice

Background: In 2020 there were over 54 million older adults in the U.S. (USCB, 2020). Nearly 90% of these older adults intend to live in their homes and communities for as long as possible (AARP, 2011) although nearly 40% of individuals 65+ report having at least one disability (USCB, 2014). Community partner LiveLife Therapy Solutions (LLTS) seeks to provide assistive technology (AT) services to community-dwelling individuals of all ages, supporting their independence. They have recently branched out into the aging sector. Continued education can aid practitioners in use of evidence-based practice (EBP) to support older adults (Stetler et al., 2014). Purpose: The purpose of this capstone project was to use education to aid interdisciplinary practitioners in use of EBP relating to the support of community-dwelling older adults through use of AT. Methods: After a review of evidence-based literature, nine educational modules were created. Five of the nine modules were presented to LLTS consultants using a combination of synchronous and asynchronous methods. The effectiveness of the education was then determined based upon analysis of pre/posttests. Results: Participants (n=16) rated education highly and education-specific learning was seen during pretest/posttest comparison. Greater changes in practitioner confidence were seen after synchronous presentation than asynchronous. Areas of key learning, proposed future topic areas, and suggestions for improvement were also identified. Implications: Findings supported that education, especially when synchronous, can help support learning and EBP. Continuing education can be used to support professionals at LLTS and beyond in the field of AT and older adults, but more research is needed on AT use with older adults

Training in Trauma-Informed Care (TIC): An Evidence-Based Practice Project

This Evidence-Based Practice (EBP) project examined the following question: What are the characteristics and effectiveness of trauma-informed care (TIC) training on the performance of health professionals and organizations who provide services to populations who have experienced trauma (traumatic events or ACE)

Genetic Associations and Differential mRNA Expression Levels of Host Genes Suggest a Viral Trigger for Endemic Pemphigus Foliaceus

The long search for the environmental trigger of the endemic pemphigus foliaceus (EPF, fogo selvagem) has not yet resulted in any tangible findings. Here, we searched for genetic asso ciations and the differential expression of host genes involved in early viral infections and innate antiviral defense. Genetic variants could alter the structure, expression sites, or levels of the gene products, impacting their functions. By analyzing 3063 variants of 166 candidate genes in 227 EPF patients and 194 controls, we found 12 variants within 11 genes associated with differential suscepti bility (p < 0.005) to EPF. The products of genes TRIM5, TPCN2, EIF4E, EIF4E3, NUP37, NUP50, NUP88, TPR, USP15, IRF8, and JAK1 are involved in different mechanisms of viral control, for example, the regulation of viral entry into the host cell or recognition of viral nucleic acids and proteins. Only two of nine variants were also associated in an independent German cohort of sporadic PF (75 patients, 150 controls), aligning with our hypothesis that antiviral host genes play a major role in EPF due to a specific virus–human interaction in the endemic region. Moreover, CCL5, P4HB, and APOBEC3G mRNA levels were increased (p < 0.001) in CD4+ T lymphocytes of EPF patients. Because there is limited or no evidence that these genes are involved in autoimmunity, their crucial role in antiviral responses and the associations that we observed support the hypothesis of a viral trigger for EPF, presumably a still unnoticed flavivirus. This work opens new frontiers in searching for the trigger of EPF, with the potential to advance translational research that aims for disease prevention and treatment

Recombinant Lysyl Oxidase Propeptide Protein Inhibits Growth and Promotes Apoptosis of Pre-Existing Murine Breast Cancer Xenografts

Lysyl oxidase propeptide (LOX-PP) ectopic overexpression inhibits the growth of cancer xenografts. Here the ability and mode of action of purified recombinant LOX-PP (rLOX-PP) protein to inhibit the growth of pre-existing xenografts was determined. Experimental approaches employed were direct intratumoral injection (i.t.) of rLOX-PP protein into murine breast cancer NF639 xenografts, and application of a slow release formulation of rLOX-PP implanted adjacent to tumors in NCR nu/nu mice (n = 10). Tumors were monitored for growth, and after sacrifice were subjected to immunohistochemical and Western blot analyses for several markers of proliferation, apoptosis, and for rLOX-PP itself. Direct i.t. injection of rLOX-PP significantly reduced tumor volume on days 20, 22 and 25 and tumor weight at harvest on day 25 by 30% compared to control. Implantation of beads preloaded with 35 micrograms rLOX-PP (n = 10) in vivo reduced tumor volume and weight at sacrifice when compared to empty beads (p<0.05). A 30% reduction of tumor volume on days 22 and 25 (p<0.05) and final tumor weight on day 25 (p<0.05) were observed with a reduced tumor growth rate of 60% after implantation. rLOX-PP significantly reduced the expression of proliferation markers and Erk1/2 MAP kinase activation, while prominent increases in apoptosis markers were observed. rLOX-PP was detected by immunohistochemistry in harvested rLOX-PP tumors, but not in controls. Data provide pre-clinical findings that support proof of principle for the therapeutic anti-cancer potential of rLOX-PP protein formulations

Finishing the euchromatic sequence of the human genome

The sequence of the human genome encodes the genetic instructions for human physiology, as well as rich information about human evolution. In 2001, the International Human Genome Sequencing Consortium reported a draft sequence of the euchromatic portion of the human genome. Since then, the international collaboration has worked to convert this draft into a genome sequence with high accuracy and nearly complete coverage. Here, we report the result of this finishing process. The current genome sequence (Build 35) contains 2.85 billion nucleotides interrupted by only 341 gaps. It covers ∼99% of the euchromatic genome and is accurate to an error rate of ∼1 event per 100,000 bases. Many of the remaining euchromatic gaps are associated with segmental duplications and will require focused work with new methods. The near-complete sequence, the first for a vertebrate, greatly improves the precision of biological analyses of the human genome including studies of gene number, birth and death. Notably, the human enome seems to encode only 20,000-25,000 protein-coding genes. The genome sequence reported here should serve as a firm foundation for biomedical research in the decades ahead

The use of the Knob-Socket model in the synthesis and expression of a novel protein, Star1.0

The knob-socket model is a code to describe how proteins interact to form tertiary structures. The basis of the knob-socket model includes a one amino acid residue knob from one piece of secondary structure packing into a three amino acid residue socket from another piece of secondary structure. Sockets can be described as free (disfavoring knob packing), or filled (favoring knob packing) and are given these designations depending on their three amino acid composition. A propensity library was developed and gives the frequency at which each socket is found to be either free or filled based on data from the PDB database. We aim to show that the knob socket model and the socket propensity library can be used to predict protein secondary structure, and therefore be used in protein design. This idea was used in the de novo design of the STAR1.0 protein. The STAR1.0 protein was designed with a unique five alpha-helical structure in the shape of a five pointed star. The sequence was developed using the alpha-helical propensity library and was further optimized for expression in E. coli. The STAR1.0 protein was expressed and purified, and then secondary structure was analyzed using circular dichroism spectroscopy. STAR1.0 is now being expressed on a large scale and purified in order to produce a high concentration of protein for x-ray crystallography, which will determine if the structure matches the theoretical five pointed star shape

The use of the Knob-Socket model in the synthesis and expression of a novel protein, Star1.0

The knob-socket model is a code to describe how proteins interact to form tertiary structures. The basis of the knob-socket model includes a one amino acid residue knob from one piece of secondary structure packing into a three amino acid residue socket from another piece of secondary structure. Sockets can be described as free (disfavoring knob packing), or filled (favoring knob packing) and are given these designations depending on their three amino acid composition. A propensity library was developed and gives the frequency at which each socket is found to be either free or filled based on data from the PDB database. We aim to show that the knob socket model and the socket propensity library can be used to predict protein secondary structure, and therefore be used in protein design. This idea was used in the de novo design of the STAR1.0 protein. The STAR1.0 protein was designed with a unique five alpha-helical structure in the shape of a five pointed star. The sequence was developed using the alpha-helical propensity library and was further optimized for expression in E. coli. The STAR1.0 protein was expressed and purified, and then secondary structure was analyzed using circular dichroism spectroscopy. STAR1.0 is now being expressed on a large scale and purified in order to produce a high concentration of protein for x-ray crystallography, which will determine if the structure matches the theoretical five pointed star shape

Application of the Knob Socket Model to predict changes in alpha helical structure

The Knob Socket (KS) model is a 4 amino acid motif that describes the way a protein will fold and pack its residues to form tertiary structure. The model includes a one amino acid residue knob from one secondary structure that packs into a three amino acid residue socket of another secondary structure. The α-helical sockets can be placed into three different categories: (1) free, unpacked and favoring intra-helical interactions, (2) filled, packed with a knob, and favoring inter-helical interactions, and (3) non, unpacked and disfavoring α-helical structure. Data within the Protein Data Bank was used to develop propensity libraries for each type of secondary structures. An α-helical propensity library was used to determine the relative frequency in which specific amino acid composition of sockets were free or filled. From this library and use of the KS model, a novel anti-parallel α-helical homodimer, KSα1.1, was designed. A single point mutation in the KSα1.1 sequence was incorporated in order to change the propensities of the surrounding six sockets and named a ‘hexagon.’ Values calculated from the difference between the total socket propensities for KSα1.1 and its corresponding mutated versions were used to predict changes in alpha helical content. Negative values corresponded to a predicted decrease in alpha helical content whereas positive values corresponded to a predicted increase in alpha helical content. Point mutations were made in the KSα1.1 sequence through the use of site-directed mutagenesis. To obtain high amounts of the desired mutated protein, plasmid vectors containing the specific point mutations in KSα1.1 sequence were transformed and expressed in E.coli. The transformed cells were induced for protein expression with Isopropyl β-D-1-thiogalactopyranoside (IPTG) and purified via column chromatography. The mutated versions of KSα1.1 were analyzed via circular dichroism (CD) spectroscopy to confirm predictions made using the KS model and propensity libraries. Deconvolutions were used to analyze the CD graphs and determine the percent content of alpha helix, beta sheet, and random coil structures. Mutant KSα1.1 proteins were compared to wild-type KSα1.1 protein in order to analyze changes in higher ordered protein packing and alpha helical structure

Application of the Knob Socket Model to predict changes in alpha helical structure

The Knob Socket (KS) model is a 4 amino acid motif that describes the way a protein will fold and pack its residues to form tertiary structure. The model includes a one amino acid residue knob from one secondary structure that packs into a three amino acid residue socket of another secondary structure. The α-helical sockets can be placed into three different categories: (1) free, unpacked and favoring intra-helical interactions, (2) filled, packed with a knob, and favoring inter-helical interactions, and (3) non, unpacked and disfavoring α-helical structure. Data within the Protein Data Bank was used to develop propensity libraries for each type of secondary structures. An α-helical propensity library was used to determine the relative frequency in which specific amino acid composition of sockets were free or filled. From this library and use of the KS model, a novel anti-parallel α-helical homodimer, KSα1.1, was designed. A single point mutation in the KSα1.1 sequence was incorporated in order to change the propensities of the surrounding six sockets and named a ‘hexagon.’ Values calculated from the difference between the total socket propensities for KSα1.1 and its corresponding mutated versions were used to predict changes in alpha helical content. Negative values corresponded to a predicted decrease in alpha helical content whereas positive values corresponded to a predicted increase in alpha helical content. Point mutations were made in the KSα1.1 sequence through the use of site-directed mutagenesis. To obtain high amounts of the desired mutated protein, plasmid vectors containing the specific point mutations in KSα1.1 sequence were transformed and expressed in E.coli. The transformed cells were induced for protein expression with Isopropyl β-D-1-thiogalactopyranoside (IPTG) and purified via column chromatography. The mutated versions of KSα1.1 were analyzed via circular dichroism (CD) spectroscopy to confirm predictions made using the KS model and propensity libraries. Deconvolutions were used to analyze the CD graphs and determine the percent content of alpha helix, beta sheet, and random coil structures. Mutant KSα1.1 proteins were compared to wild-type KSα1.1 protein in order to analyze changes in higher ordered protein packing and alpha helical structure