NIAGADS: A Comprehensive National Data Repository for Alzheimer's Disease and Related Dementia Genetics and Genomics Research.

NIAGADS is the National Institute on Aging (NIA) designated national data repository for human genetics research on Alzheimer's Disease and related dementia (ADRD). NIAGADS maintains a high-quality data collection for ADRD genetic/genomic research and supports genetics data production and analysis. NIAGADS hosts whole genome and exome sequence data from the Alzheimer's Disease Sequencing Project (ADSP) and other genotype/phenotype data, encompassing 209,000 samples. NIAGADS shares these data with hundreds of research groups around the world via the Data Sharing Service, a FISMA moderate compliant cloud-based platform that fully supports the NIH Genome Data Sharing Policy. NIAGADS Open Access consists of multiple knowledge bases with genome-wide association summary statistics and rich annotations on the biological significance of genetic variants and genes across the human genome. NIAGADS stands as a keystone in promoting collaborations to advance the understanding and treatment of Alzheimer's disease.

Different Sensory Information Is Used for State Estimation when Stationary or Moving.

The accurate estimation of limb state is necessary for movement planning and execution. While state estimation requires both feedforward and feedback information, we focus here on the latter. Prior literature has shown that integrating visual and proprioceptive feedback improves estimates of static limb position. However, differences in visual and proprioceptive feedback delays suggest that multisensory integration could be disadvantageous when the limb is moving. We formalized this hypothesis by modeling feedback-based state estimation using the long-standing maximum likelihood estimation model of multisensory integration, which we updated to account for sensory delays. Our model predicted that the benefit of multisensory integration was largely lost when the limb was passively moving. We tested this hypothesis in a series of experiments in human subjects that compared the degree of interference created by discrepant visual or proprioceptive feedback when estimating limb position either statically at the end of the movement or dynamically at movement midpoint. In the static case, we observed significant interference: discrepant feedback in one modality systematically biased sensory estimates based on the other modality. However, no interference was seen in the dynamic case: participants could ignore sensory feedback from one modality and accurately reproduce the motion indicated by the other modality. Together, these findings suggest that the sensory feedback used to compute a state estimate differs depending on whether the limb is stationary or moving. While the former may tend toward multimodal integration, the latter is more likely to be based on feedback from a single sensory modality.

Different sensory information is used for state estimation when stationary or moving.

Accurate estimation of limb state is necessary for movement planning and execution. State estimation requires both feedforward and feedback information; here we focus on the latter. Prior literature has shown that integrating visual and proprioceptive feedback improve estimates of static limb position. However, differences in visual and proprioceptive feedback delays suggest that multisensory integration could be disadvantageous when the limb is moving. To investigate multisensory integration in different passive movement contexts, we compared the degree of interference created by discrepant visual or proprioceptive feedback when estimating the position of the limb either statically at the end of the movement or dynamically at movement midpoint. In the static context, we observed idiosyncratic interference: discrepant proprioceptive feedback significantly interfered with reports of the visual target location, leading to a bias of the reported position toward the proprioceptive cue. In the dynamic context, no interference was seen: participants could ignore sensory feedback from one modality and accurately reproduce the motion indicated by the other modality. We modeled feedback-based state estimation by updating the longstanding maximum likelihood estimation model of multisensory integration to account for sensory delays. Consistent with our behavioral results, the model showed that the benefit of multisensory integration was largely lost when the limb was passively moving. Together, these findings suggest that the sensory feedback used to compute a state estimate differs depending on whether the limb is stationary or moving. While the former may tend toward multimodal integration, the latter is more likely to be based on feedback from a single sensory modality.

Laser texturing of additively manufactured implants: A tool to programme biological response.

The advent of additive manufacturing (AM) is rapidly shaping healthcare technologies pushing forward personalisation and enhanced implant functionalisation to improve clinical outcomes. AM techniques such as powder bed fusion (PBF) have been adopted despite the need to modify the as-built surface post manufacture. Medical device manufacturers have focused their efforts on refining various physical and chemical surface finishing approaches, however there is little consensus and some methods risk geometry alteration or contamination. This has led to a growing interest in laser texturing technologies to engineer the device surface. Herein, several bioinspired micro and nano textures were applied to laser PBF Ti-6Al-V4 substrates to alter physicochemical properties and in-turn we sought to understand what influences these alterations had on a human osteosarcoma cell line (MG63). Significant variations in roughness and time dependent contact angles were revealed between different patterns provide a tool to elicit desired biological responses. All surface treatments effectively enhanced early cell behaviour and in particular coverage was increased for the micro-textures. Influence of the patterns on cell differentiation was less consistent with alkaline phosphatase content increased only for the channel, grid and dual textures. While long term (21 days) mineralisation was found to be significantly enhanced in grids, dual, triangles and shark skin textures. Further regression analysis of all physicochemical and biological variables indicated that several properties should be used to strongly correlate cell behaviour, resulting in 82 % of the 21 day mineralisation dataset explained through a combination of roughness kurtosis and glycerol contact angle. Overall, this manuscript demonstrates the ability of laser texturing to offer tailored cell-surface interactions, which can be tuned to offer a tool to drive functional customisation of anatomically customised medical devices.

Stakeholder Perspectives on the Current and Future of Additive Manufacturing in Healthcare.

Additive manufacturing (AM) technologies have disrupted many supply chains by making new designs and functionalities possible. The opportunity to realize complex customized structures has led to significant interest within healthcare; however, full utilization critically requires the alignment of the whole supply chain. To offer insights into this process, a survey was conducted to understand the views of different medical AM stakeholders. The results highlighted an agreement between academics, designers, manufacturers, and medical experts, that personalization and design control are the main benefits of AM. Interestingly, surface finish was consistently identified as an obstacle. Nevertheless, there was a degree of acceptance that post-processing was necessary to achieve appropriate quality control. Recommendations were made for extending the use of in situ process monitoring systems to support improved reproducibility. Variations in the future vision of AM were highlighted between stakeholder groups and areas of interest for development noted for each stakeholder. Collectively, this survey indicates that medical stakeholders agree on the capabilities of AM but have different priorities for its implementation and progression. This highlights a degree of disconnection among the supply chain at a ground level; thus, collaboration on AM specific standards and enhancement of communication between stakeholders from project inception is recommended.

Surface Free Energy Dominates the Biological Interactions of Postprocessed Additively Manufactured Ti-6Al-4V.

Additive manufacturing (AM) has emerged as a disruptive technique within healthcare because of its ability to provide personalized devices; however, printed metal parts still present surface and microstructural defects, which may compromise mechanical and biological interactions. This has made physical and/or chemical postprocessing techniques essential for metal AM devices, although limited fundamental knowledge is available on how alterations in physicochemical properties influence AM biological outcomes. For this purpose, herein, powder bed fusion Ti-6Al-4V samples were postprocessed with three industrially relevant techniques: polishing, passivation, and vibratory finishing. These surfaces were thoroughly characterized in terms of roughness, chemistry, wettability, surface free energy, and surface ζ-potential. A significant increase in Staphylococcus epidermidis colonization was observed on both polished and passivated samples, which was linked to high surface free energy donor γ- values in the acid-base, γAB component. Early osteoblast attachment and proliferation (24 h) were not influenced by these properties, although increased mineralization was observed for both these samples. In contrast, osteoblast differentiation on stainless steel was driven by a combination of roughness and chemistry. Collectively, this study highlights that surface free energy is a key driver between AM surfaces and cell interactions. In particular, while low acid-base components resulted in a desired reduction in S. epidermidis colonization, this was followed by reduced mineralization. Thus, while surface free energy can be used as a guide to AM device development, optimization of bacterial and mammalian cell interactions should be attained through a combination of different postprocessing techniques.

A feasible route for the design and manufacture of customised respiratory protection through digital facial capture.

The World Health Organisation has called for a 40% increase in personal protective equipment manufacturing worldwide, recognising that frontline workers need effective protection during the COVID-19 pandemic. Current devices suffer from high fit-failure rates leaving significant proportions of users exposed to risk of viral infection. Driven by non-contact, portable, and widely available 3D scanning technologies, a workflow is presented whereby a user's face is rapidly categorised using relevant facial parameters. Device design is then directed down either a semi-customised or fully-customised route. Semi-customised designs use the extracted eye-to-chin distance to categorise users in to pre-determined size brackets established via a cohort of 200 participants encompassing 87.5% of the cohort. The user's nasal profile is approximated to a Gaussian curve to further refine the selection in to one of three subsets. Flexible silicone provides the facial interface accommodating minor mismatches between true nasal profile and the approximation, maintaining a good seal in this challenging region. Critically, users with outlying facial parameters are flagged for the fully-customised route whereby the silicone interface is mapped to 3D scan data. These two approaches allow for large scale manufacture of a limited number of design variations, currently nine through the semi-customised approach, whilst ensuring effective device fit. Furthermore, labour-intensive fully-customised designs are targeted as those users who will most greatly benefit. By encompassing both approaches, the presented workflow balances manufacturing scale-up feasibility with the diverse range of users to provide well-fitting devices as widely as possible. Novel flow visualisation on a model face is presented alongside qualitative fit-testing of prototype devices to support the workflow methodology.

Novel Hybrid Manufacturing Process of CM247LC and Multi-Material Blisks.

The study on CM247LC used the traditional approach for Near-Netshape Hot Isostatic Pressing (NNSHIP) with sacrificial low carbon steel tooling, which was built using Selective Laser Melting (SLM), to produce a shaped CM247LC blisk. The assessment of the microstructure focused on both the exterior components in order to determine the depth of the Fe-diffusion layer and on the interior microstructure. Samples were extracted from the Hot Isostatic Pressed (HIPped) components for tensile testing at both room and elevated temperatures. The components were scanned to assess the geometrical shrinkages due to Hot Isostatic Pressing (HIPping). An oversized blisk was also produced based on the measurements as a demonstrator component. In addition, a further study was carried out on a novel idea that used a solid IN718 disk in the centre of the blisk to create a multi-material component.

Reducing MRI susceptibility artefacts in implants using additively manufactured porous Ti-6Al-4V structures.

Magnetic Resonance Imaging (MRI) is critical in diagnosing post-operative complications following implant surgery and imaging anatomy adjacent to implants. Increasing field strengths and use of gradient-echo sequences have highlighted difficulties from susceptibility artefacts in scan data. Artefacts manifest around metal implants, including those made from titanium alloys, making detection of complications (e.g. bleeding, infection) difficult and hindering imaging of surrounding structures such as the brain or inner ear. Existing research focusses on post-processing and unorthodox scan sequences to better capture data around these devices. This study proposes a complementary up-stream design approach using lightweight structures produced via additive manufacturing (AM). Strategic implant mass reduction presents a potential tool in managing artefacts. Uniform specimens of Ti-6Al-4V structures, including lattices, were produced using the AM process, selective laser melting, with various unit cell designs and relative densities (3.1%-96.7%). Samples, submerged in water, were imaged in a 3T MRI system using clinically relevant sequences. Artefacts were quantified by image analysis revealing a strong linear relationship (RR2 = 0.99) between severity and relative sample density. Likewise, distortion due to slice selection errors showed a squared relationship (RR2 = 0.92) with sample density. Unique artefact features were identified surrounding honeycomb samples suggesting a complex relationship exists for larger unit cells. To demonstrate clinical utility, a honeycomb design was applied to a representative cranioplasty. Analysis revealed 10% artefact reduction compared to traditional solid material illustrating the feasibility of this approach. This study provides a basis to strategically design implants to reduce MRI artefacts and improve post-operative diagnosis capability. STATEMENT OF SIGNIFICANCE: MRI susceptibility artefacts surrounding metal implants present a clinical challenge for the diagnosis of post-operative complications relating to the implant itself or underlying anatomy. In this study for the first time we demonstrate that additive manufacturing may be exploited to create lattice structures that predictably reduce MRI image artefact severity surrounding titanium alloy implants. Specifically, a direct correlation of artefact severity, both total signal loss and distortion, with the relative material density of these functionalised materials has been demonstrated within clinically relevant MRI sequences. This approach opens the door for strategic implant design, utilising this structurally functionalised material, that may improve post-operative patient outcomes and compliments existing efforts in this area which focus on data acquisition and post-processing methods.

A pre-steady state and steady state kinetic analysis of the N-ribosyl hydrolase activity of hCD157.

hCD157 catalyzes the hydrolysis of nicotinamide riboside (NR) and nicotinic acid riboside (NAR). The release of nicotinamide or nicotinic acid from NR or NAR was confirmed by spectrophotometric, HPLC and NMR analyses. hCD157 is inactivated by a mechanism-based inhibitor, 2'-deoxy-2'-fluoro-nicotinamide arabinoside (fNR). Modification of the enzyme during the catalytic cycle by NR, NAR, or fNR increased the intrinsic protein fluorescence by approximately 50%. Pre-steady state and steady state data were used to derive a minimal kinetic scheme for the hydrolysis of NR. After initial complex formation a reversible step (360 and 30s(-1)) is followed by a slow irreversible step (0.1s(-1)) that defined the rate limiting step, or kcat. The calculated KMapp value for NR in the hydrolytic reaction is 6nM. The values of the kinetic constants suggest that one biological function of cell-surface hCD157 is to bind and slowly hydrolyze NR, possibly converting it to a ligand-activated receptor. Differences in substrate preference between hCD157 and hCD38 were rationalized through a comparison of the crystal structures of the two proteins. This comparison identified several residues in hCD157 (F108 and F173) that can potentially hinder the binding of dinucleotide substrates (NAD+).

Rapid mapping and identification of mutations in Caenorhabditis elegans by restriction site-associated DNA mapping and genomic interval pull-down sequencing.

Forward genetic screens provide a powerful approach for inferring gene function on the basis of the phenotypes associated with mutated genes. However, determining the causal mutation by traditional mapping and candidate gene sequencing is often the rate-limiting step, especially when analyzing many mutants. We report two genomic approaches for more rapidly determining the identity of the affected genes in Caenorhabditis elegans mutants. First, we report our use of restriction site-associated DNA (RAD) polymorphism markers for rapidly mapping mutations after chemical mutagenesis and mutant isolation. Second, we describe our use of genomic interval pull-down sequencing (GIPS) to selectively capture and sequence megabase-sized portions of a mutant genome. Together, these two methods provide a rapid and cost-effective approach for positional cloning of C. elegans mutant loci, and are also applicable to other genetic model systems.

A survey of new temperature-sensitive, embryonic-lethal mutations in C. elegans: 24 alleles of thirteen genes.

To study essential maternal gene requirements in the early C. elegans embryo, we have screened for temperature-sensitive, embryonic lethal mutations in an effort to bypass essential zygotic requirements for such genes during larval and adult germline development. With conditional alleles, multiple essential requirements can be examined by shifting at different times from the permissive temperature of 15°C to the restrictive temperature of 26°C. Here we describe 24 conditional mutations that affect 13 different loci and report the identity of the gene mutations responsible for the conditional lethality in 22 of the mutants. All but four are mis-sense mutations, with two mutations affecting splice sites, another creating an in-frame deletion, and one creating a premature stop codon. Almost all of the mis-sense mutations affect residues conserved in orthologs, and thus may be useful for engineering conditional mutations in other organisms. We find that 62% of the mutants display additional phenotypes when shifted to the restrictive temperature as L1 larvae, in addition to causing embryonic lethality after L4 upshifts. Remarkably, we also found that 13 out of the 24 mutations appear to be fast-acting, making them particularly useful for careful dissection of multiple essential requirements. Our findings highlight the value of C. elegans for identifying useful temperature-sensitive mutations in essential genes, and provide new insights into the requirements for some of the affected loci.

Pyrrolidinyl pyridone and pyrazinone analogues as potent inhibitors of prolyl oligopeptidase (POP).

We report the synthesis and in vitro activity of a series of novel pyrrolidinyl pyridones and pyrazinones as potent inhibitors of prolyl oligopeptidase (POP). Within this series, compound 39 was co-crystallized within the catalytic site of a human chimeric POP protein which provided a more detailed understanding of how these inhibitors interacted with the key residues within the catalytic pocket.

An intrinsic ATPase activity of phospho-MEK-1 uncoupled from downstream ERK phosphorylation.

We have developed a highly sensitive assay of MEK-mediated ATP hydrolysis by coupling the formation of ADP to NADH oxidation through the enzymes pyruvate kinase and lactate dehydrogenase. Robust ATP hydrolysis is catalyzed by phosphorylated MEK in the absence of the protein substrate ERK. This ERK-uncoupled ATPase activity is dependent on the phosphorylation status of MEK and is abrogated by the selective MEK kinase inhibitor U0126. ADP production is concomitant with Raf-mediated phosphorylation of MEK. Based on this finding, a coupled Raf/MEK assay is developed for measuring the Raf activity. A kinetic treatment derived under steady-state assumptions is presented for the analysis of the reaction progress curve generated by this coupled assay. We have shown that inhibitory potency of selective Raf inhibitors can be determined accurately by this assay.