Publications by authors named "Andrew West"

138 Publications

Sex-based differences in the activation of peripheral blood monocytes in early Parkinson disease.

NPJ Parkinsons Dis 2021 Apr 13;7(1):36. Epub 2021 Apr 13.

Alabama Morris K. Udall Center of Excellence in Parkinson's Disease Research, University of Alabama at Birmingham, 1719 Sixth Ave. South, Birmingham, AL, USA.

Increasing evidence supports the role of brain and systemic inflammation in the etiology of Parkinson disease (PD). We used gene expression profiling to examine the activation state of peripheral blood monocytes in 18 patients with early, untreated PD and 16 healthy control (HC) subjects. Monocytes were isolated by negative selection, and gene expression studied by RNA-seq and gene set enrichment analysis. A computational model that incorporated case/control status, sex, and the interaction between case/control status and sex was utilized. We found that there was a striking effect of sex on monocyte gene expression. There was inflammatory activation of monocytes in females with PD, with enrichment of gene sets associated with interferon gamma stimulation. In males, the activation patterns were more heterogeneous. These data point to the importance of systemic monocyte activation in PD, and the importance of studies which examine the differential effects of sex on pathophysiology of the disease.
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http://dx.doi.org/10.1038/s41531-021-00180-zDOI Listing
April 2021

"OMG, I Get Like 100 Teaspoons of Sugar a Day!" Rural Teens' Grasp of Their Beverage Consumption Habits.

Int Q Community Health Educ 2021 Mar 22:272684X211004928. Epub 2021 Mar 22.

Department of Communication and Journalism, University of New Mexico, Albuquerque, New Mexico, United States.

As part of a campaign to encourage healthier beverage consumption in a rural New Mexico high school, we wanted to understand students' perceptions of their habits, and the associated health risks and benefits surrounding water and sugar-sweetened beverage consumption to influence future messaging to change behavior. We conducted a posthoc qualitative analysis of 27 student interviews from the program evaluation with both students who participated in the campaign and those who did not. Pre-campaign, students appeared largely unaware of the health risks posed by their beverage consumption habits, lacking any knowledge of nutritional recommendations, water recommendations, or of the sugar levels in products. When informed of the risks, students expressed concern for themselves, their family members, and friends, indicated a desire to make significant changes, and reported making changes for themselves, and educating others regarding the risks. Given the large amounts of money spent and concentrated efforts focused on marketing SSBs to teens, it is critically important to be educating teens and engaging them in behavior change strategies. These data can inform future strategies to improve teen health behaviors and encourage teens to become family health messengers for obesity and diabetes prevention.
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http://dx.doi.org/10.1177/0272684X211004928DOI Listing
March 2021

Age-associated insolubility of parkin in human midbrain is linked to redox balance and sequestration of reactive dopamine metabolites.

Acta Neuropathol 2021 05 10;141(5):725-754. Epub 2021 Mar 10.

Program in Neuroscience, Ottawa Hospital Research Institute, Ottawa, ON, Canada.

The mechanisms by which parkin protects the adult human brain from Parkinson disease remain incompletely understood. We hypothesized that parkin cysteines participate in redox reactions and that these are reflected in its posttranslational modifications. We found that in post mortem human brain, including in the Substantia nigra, parkin is largely insoluble after age 40 years; this transition is linked to its oxidation, such as at residues Cys95 and Cys253. In mice, oxidative stress induces posttranslational modifications of parkin cysteines that lower its solubility in vivo. Similarly, oxidation of recombinant parkin by hydrogen peroxide (HO) promotes its insolubility and aggregate formation, and in exchange leads to the reduction of HO. This thiol-based redox activity is diminished by parkin point mutants, e.g., p.C431F and p.G328E. In prkn-null mice, HO levels are increased under oxidative stress conditions, such as acutely by 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine toxin exposure or chronically due to a second, genetic hit; HO levels are also significantly increased in parkin-deficient human brain. In dopamine toxicity studies, wild-type parkin, but not disease-linked mutants, protects human dopaminergic cells, in part through lowering HO. Parkin also neutralizes reactive, electrophilic dopamine metabolites via adduct formation, which occurs foremost at the primate-specific residue Cys95. Further, wild-type but not p.C95A-mutant parkin augments melanin formation in vitro. By probing sections of adult, human midbrain from control individuals with epitope-mapped, monoclonal antibodies, we found specific and robust parkin reactivity that co-localizes with neuromelanin pigment, frequently within LAMP-3/CD63 lysosomes. We conclude that oxidative modifications of parkin cysteines are associated with protective outcomes, which include the reduction of HO, conjugation of reactive dopamine metabolites, sequestration of radicals within insoluble aggregates, and increased melanin formation. The loss of these complementary redox effects may augment oxidative stress during ageing in dopamine-producing cells of mutant PRKN allele carriers, thereby enhancing the risk of Parkinson's-linked neurodegeneration.
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http://dx.doi.org/10.1007/s00401-021-02285-4DOI Listing
May 2021

Heterogeneity in α-synuclein fibril activity correlates to disease phenotypes in Lewy body dementia.

Acta Neuropathol 2021 04 28;141(4):547-564. Epub 2021 Feb 28.

Department of Pharmacology and Cancer Biology, Duke Center for Neurodegeneration Research, Duke University, 3 Genome Court, Durham, NC, 27710, USA.

α-Synuclein aggregation underlies pathological changes in Lewy body dementia. Recent studies highlight structural variabilities associated with α-synuclein aggregates in patient populations. Here, we develop a quantitative real-time quaking-induced conversion (qRT-QuIC) assay to measure permissive α-synuclein fibril-templating activity in tissues and cerebrospinal fluid (CSF). The assay is anchored through reference panels of stabilized ultra-short fibril particles. In humanized α-synuclein transgenic mice, qRT-QuIC identifies differential levels of fibril activity across the brain months before the deposition of phosphorylated α-synuclein in susceptible neurons. α-Synuclein fibril activity in cortical brain extracts from dementia with Lewy bodies (DLB) correlates with activity in matched ventricular CSF. Elevated α-synuclein fibril activity in CSF corresponds to reduced survival in DLB. α-Synuclein fibril particles amplified from cases with high fibril activity show superior templating in the formation of new inclusions in neurons relative to the same number of fibril particles amplified from DLB cases with low fibril activity. Our results highlight a previously unknown broad heterogeneity of fibril-templating activities in DLB that may contribute to disease phenotypes. We predict that quantitative assessments of fibril activities in CSF that correlate to fibril activities in brain tissue will help stratify patient populations as well as measure therapeutic responses to facilitate the development of α-synuclein-targeted therapeutics.
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http://dx.doi.org/10.1007/s00401-021-02288-1DOI Listing
April 2021

Identification of LRRK2 Missense Variants in the Accelerating Medicines Partnership Parkinson's Disease Cohort.

Hum Mol Genet 2021 Feb 27. Epub 2021 Feb 27.

Duke Center for Neurodegeneration and Neurotherapeutics Research, Departments of Pharmacology and Cancer Biology, and Neurology, Duke University, Durham, North Carolina, 27710 USA.

Pathogenic missense variants in the leucine-rich repeat kinase 2 (LRRK2) gene have been identified through linkage analysis in familial Parkinson disease (PD). Subsequently, other missense variants with lower effect sizes on PD risk have emerged, as well as non-coding polymorphisms (e.g. rs76904798) enriched in PD cases in genome-wide association studies. Here we leverage recent whole-genome sequences from the Accelerating Medicines Partnership-Parkinson's Disease (AMP-PD) and the Genome Aggregation (gnomAD) databases to characterize novel missense variants in LRRK2 and explore their relationships with known pathogenic and PD-linked missense variants. Using a computational prediction tool that successfully classifies known pathogenic LRRK2 missense variants, we describe an online web-based resource that catalogs characteristics of over twelve-hundred LRRK2 missense variants of unknown significance. Novel high-pathogenicity scoring variants, some identified exclusively in PD cases, tightly cluster within the ROC-COR-Kinase domains. Structure-function predictions support that some of these variants exert gain-of-function effects with respect to LRRK2 kinase activity. In AMP-PD participants, all p.R1441G carriers (N = 89) are also carriers of the more common PD-linked variant p.M1646T. In addition, nearly all carriers of the PD-linked p.N2081D missense variant are also carriers of the LRRK2 PD-risk variant rs76904798. These results provide a compendium of LRRK2 missense variants and how they associate with one another. While the pathogenic p.G2019S variant is by far the most frequent high-pathogenicity scoring variant, our results suggest that ultra-rare missense variants may have an important cumulative impact in increasing the number of individuals with LRRK2-linked PD.
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http://dx.doi.org/10.1093/hmg/ddab058DOI Listing
February 2021

Genetic background influences LRRK2-mediated Rab phosphorylation in the rat brain.

Brain Res 2021 May 15;1759:147372. Epub 2021 Feb 15.

Duke Center for Neurodegeneration Research, Department of Pharmacology and Cancer Biology, Duke University, Durham, NC, USA. Electronic address:

Pathogenic missense mutations in the leucine-rich repeat kinase 2 gene, encoding LRRK2, results in the upregulation of Rab10 and Rab12 phosphorylation in different cells and tissues. Here, we evaluate levels of the LRRK2 kinase substrates pT73-Rab10 and pS106-Rab12 proteins in rat brain tissues from different genetic backgrounds. Whereas lines of Sprague Dawley rats have equivalent levels of pT73-Rab10 and pS106-Rab12 similar to Lrrk2 knockout rats, Long-Evans rats have levels of pT73-Rab10 and pS106-Rab12 comparable to G2019S-LRRK2 BAC transgenic rats. Strong LRRK2 kinase inhibitors are ineffective at reducing pT73-Rab10 and pS106-Rab12 levels in the Sprague Dawley rats, but potently reduce pT73-Rab10 and pS106-Rab12 levels in Long-Evans rats. Oral administration of the PFE-360 LRRK2 kinase inhibitor fails to provide neuroprotection from dopaminergic neurodegeneration caused by rAAV2/1-mediated overexpression of A53T-αsynuclein in Sprague Dawley rats. These results highlight substantial differences in LRRK2-mediated Rab10 and Rab12 phosphorylation in commonly utilized rat genetic backgrounds and suggest LRRK2 may not play a central role in Rab phosphorylation or mutant αsynuclein toxicity in Sprague Dawley rats.
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http://dx.doi.org/10.1016/j.brainres.2021.147372DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8006086PMC
May 2021

Genetic and Environmental Factors in Parkinson's Disease Converge on Immune Function and Inflammation.

Mov Disord 2021 01 14;36(1):25-36. Epub 2020 Dec 14.

Departments of Neuroscience and Neurology, Center for Translational Research in Neurodegenerative Disease, College of Medicine, University of Florida, Gainesville, Florida, USA.

Idiopathic Parkinson's disease (iPD) is a movement disorder characterized by the degeneration of dopaminergic neurons and aggregation of the protein α-synuclein. Patients with iPD vary in age of symptom onset, rate of progression, severity of motor and non-motor symptoms, and extent of central and peripheral inflammation. Genetic and environmental factors are believed to act synergistically in iPD pathogenesis. We propose that environmental factors (pesticides and infections) increase the risk for iPD via the immune system and that the role of PD risk genes in immune cells is worthy of investigation. This review highlights the major PD-relevant genes expressed in immune cells and key environmental factors that activate immune cells and, alone or in combination with other factors, may contribute to iPD pathogenesis. By reviewing these interactions, we seek to enable the future development of immunomodulatory approaches to prevent or delay onset of iPD. © 2020 International Parkinson and Movement Disorder Society.
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http://dx.doi.org/10.1002/mds.28411DOI Listing
January 2021

Exosome markers of LRRK2 kinase inhibition.

NPJ Parkinsons Dis 2020 Nov 13;6(1):32. Epub 2020 Nov 13.

Department of Pharmacology and Cancer Biology, Duke Center for Neurodegeneration Research, Duke University, Durham, NC, USA.

Hyper-activated LRRK2 is linked to Parkinson's disease susceptibility and progression. Quantitative measures of LRRK2 inhibition, especially in the brain, maybe critical in the development of successful LRRK2-targeting therapeutics. In this study, two different brain-penetrant and selective LRRK2 small-molecule kinase inhibitors (PFE-360 and MLi2) were orally administered to groups of cynomolgus macaques. Proposed pharmacodynamic markers in exosomes from urine and cerebrospinal fluid (CSF) were compared to established markers in peripheral blood mononuclear cells (PBMCs). LRRK2 kinase inhibition led to reductions in exosome-LRRK2 protein and the LRRK2-substrate pT73-Rab10 in urine, as well as reduced exosome-LRRK2 and autophosphorylated pS1292-LRRK2 protein in CSF. We propose orthogonal markers for LRRK2 inhibition in urine and CSF can be used in combination with blood markers to non-invasively monitor the potency of LRRK2-targeting therapeutics.
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http://dx.doi.org/10.1038/s41531-020-00138-7DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7666125PMC
November 2020

Features of scholarly practice in health care professionals: A scoping review protocol.

Can J Respir Ther 2020 10;56:38-41. Epub 2020 Sep 10.

School of Physical and Occupational Therapy, McGill University, Montréal, QC, Canada.

Introduction: Health care professionals are expected to embrace and enact the scholarly practitioner role. Scholarly practitioners demonstrate a lifelong commitment to excellence in practice through continuous learning, engagement in evidence-informed decision-making, contributions to scholarship, and knowledge translation. However, the specific features and requirements associated with this role are not uniform. The absence of well-defined and delineated conceptualizations of scholarly practice and the scarcity of empirical research on how scholarly practice is operationalized contribute to a lack of a shared understanding of this complex role.

Aim: The purpose of this scoping review is to map the breadth and depth of the literature on what is known about scholarly practice in licensed health care professionals.

Methods: Arksey and O'Malley's 6-stage scoping review framework will be used to examine the breadth and depth of the literature on the definitions and conceptualizations of the scholar role in health care professionals. We will conduct a comprehensive search from inception to present in MEDLINE (Ovid), EMBASE (Ovid), and CINAHL using scholarly practitioner terms and related synonyms, including a grey literature search. Following a calibration exercise, two independent reviewers will screen retrieved papers for inclusion and extract relevant data. Included papers will: (i) explore, describe, or define scholarly practice, scholar or scholarly practitioner, and/or related concepts in the licensed health care professionals; (ii) be conceptual and/or theoretical in nature; (iii) use quantitative, qualitative, or mixed methodologies; and (iv) be published in English or French. Numeric and thematic analysis will characterize the data and address the research objectives.
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7485963PMC
September 2020

Pharmacodynamic Biomarkers for Emerging LRRK2 Therapeutics.

Front Neurosci 2020 6;14:807. Epub 2020 Aug 6.

Duke Center for Neurodegeneration Research, Departments of Pharmacology and Cancer Biology, Neurology, and Neurobiology, Duke University, Durham, NC, United States.

Genetic studies have identified variants in the gene as important components of Parkinson's disease (PD) pathobiology. Biochemical and emergent biomarker studies have coalesced around LRRK2 hyperactivation in disease. Therapeutics that diminish LRRK2 activity, either with small molecule kinase inhibitors or anti-sense oligonucleotides, have recently advanced to the clinic. Historically, there have been few successes in the development of therapies that might slow or halt the progression of neurodegenerative diseases. Over the past few decades of biomedical research, retrospective analyses suggest the broad integration of informative biomarkers early in development tends to distinguish successful pipelines from those that fail early. Herein, we discuss the biomarker regulatory process, emerging LRRK2 biomarker candidates, assays, underlying biomarker biology, and clinical integration.
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http://dx.doi.org/10.3389/fnins.2020.00807DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7438883PMC
August 2020

LRRK2 and Rab10 coordinate macropinocytosis to mediate immunological responses in phagocytes.

EMBO J 2020 10 27;39(20):e104862. Epub 2020 Aug 27.

Duke Center for Neurodegeneration Research, Department of Pharmacology and Cancer Biology, Duke University, Durham, NC, USA.

Genetic variation in LRRK2 associates with the susceptibility to Parkinson's disease, Crohn's disease, and mycobacteria infection. High expression of LRRK2 and its substrate Rab10 occurs in phagocytic cells in the immune system. In mouse and human primary macrophages, dendritic cells, and microglia-like cells, we find that Rab10 specifically regulates a specialized form of endocytosis known as macropinocytosis, without affecting phagocytosis or clathrin-mediated endocytosis. LRRK2 phosphorylates cytoplasmic PI(3,4,5)P3-positive GTP-Rab10, before EEA1 and Rab5 recruitment to early macropinosomes occurs. Macropinosome cargo in macrophages includes CCR5, CD11b, and MHCII, and LRRK2-phosphorylation of Rab10 potently blocks EHBP1L1-mediated recycling tubules and cargo turnover. EHBP1L1 overexpression competitively inhibits LRRK2-phosphorylation of Rab10, mimicking the effects of LRRK2 kinase inhibition in promoting cargo recycling. Both Rab10 knockdown and LRRK2 kinase inhibition potently suppress the maturation of macropinosome-derived CCR5-loaded signaling endosomes that are critical for CCL5-induced immunological responses that include Akt activation and chemotaxis. These data support a novel signaling axis in the endolysosomal system whereby LRRK2-mediated Rab10 phosphorylation stalls vesicle fast recycling to promote PI3K-Akt immunological responses.
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http://dx.doi.org/10.15252/embj.2020104862DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7560233PMC
October 2020

Dopaminergic neurodegeneration induced by Parkinson's disease-linked G2019S LRRK2 is dependent on kinase and GTPase activity.

Proc Natl Acad Sci U S A 2020 07 6;117(29):17296-17307. Epub 2020 Jul 6.

Center for Neurodegenerative Science, Van Andel Research Institute, Grand Rapids, MI 49503;

Mutations in () are the most common cause of late-onset, autosomal-dominant familial Parkinson's disease (PD). LRRK2 functions as both a kinase and GTPase, and PD-linked mutations are known to influence both enzymatic activities. While PD-linked LRRK2 mutations can commonly induce neuronal damage in culture models, the mechanisms underlying these pathogenic effects remain uncertain. Rodent models containing familial LRRK2 mutations often lack robust PD-like neurodegenerative phenotypes. Here, we develop a robust preclinical model of PD in adult rats induced by the brain delivery of recombinant adenoviral vectors with neuronal-specific expression of human LRRK2 harboring the most common G2019S mutation. In this model, G2019S LRRK2 induces the robust degeneration of substantia nigra dopaminergic neurons, a pathological hallmark of PD. Introduction of a stable kinase-inactive mutation or administration of the selective kinase inhibitor, PF-360, attenuates neurodegeneration induced by G2019S LRRK2. Neuroprotection provided by pharmacological kinase inhibition is mediated by an unusual mechanism involving the robust destabilization of human LRRK2 protein in the brain relative to endogenous LRRK2. Our study further demonstrates that G2019S LRRK2-induced dopaminergic neurodegeneration critically requires normal GTPase activity, as hypothesis-testing mutations that increase GTP hydrolysis or impair GTP-binding activity provide neuroprotection although via distinct mechanisms. Taken together, our data demonstrate that G2019S LRRK2 induces neurodegeneration in vivo via a mechanism that is dependent on kinase and GTPase activity. Our study provides a robust rodent preclinical model of -linked PD and nominates kinase inhibition and modulation of GTPase activity as promising disease-modifying therapeutic targets.
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http://dx.doi.org/10.1073/pnas.1922184117DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7382283PMC
July 2020

Veterinary Educator Teaching and Scholarship (VETS): A Case Study of a Multi-Institutional Faculty Development Program to Advance Teaching and Learning.

J Vet Med Educ 2020 Oct 11;47(5):632-646. Epub 2020 Jun 11.

Content expertise in basic science and clinical disciplines does not assure proficiency in teaching. Faculty development to improve teaching and learning is essential for the advancement of veterinary education. The Consortium of West Region Colleges of Veterinary Medicine established the Regional Teaching Academy (RTA) with the focus of "Making Teaching Matter." The objective of the RTA's first effort, the Faculty Development Initiative (FDI), was to develop a multi-institutional faculty development program for veterinary educators to learn about and integrate effective teaching methods. In 2016, the Veterinary Educator Teaching and Scholarship (VETS) program was piloted at Oregon State University's College of Veterinary Medicine. This article uses a case study approach to program evaluation of the VETS program. We describe the VETS program, participants' perceptions, participants' teaching method integration, and lessons learned. A modified Kirkpatrick Model (MKM) was used to categorize program outcomes and impact. Quantitative data are presented as descriptive statistics, and qualitative data are presented as the themes that emerged from participant survey comments and post-program focus groups. Results indicated outcomes and impacts that included participants' perceptions of the program, changes in participant attitude toward teaching and learning, an increase in the knowledge level of participants, self-reported changes in participant behaviors, and changes in practices and structure at the college level. Lessons learned indicate that the following are essential for program success: (1) providing institutional and financial support; (2) creating a community of practice (COP) of faculty development facilitators, and (3) developing a program that addresses the needs of faculty and member institutions.
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http://dx.doi.org/10.3138/jvme-2019-0089DOI Listing
October 2020

Parkinson disease and the immune system - associations, mechanisms and therapeutics.

Nat Rev Neurol 2020 Jun 24;16(6):303-318. Epub 2020 Apr 24.

Parkinson's Disease Center and Movement Disorders Clinic, Department of Neurology, Baylor College of Medicine, Houston, TX, USA.

Multiple lines of evidence indicate that immune system dysfunction has a role in Parkinson disease (PD); this evidence includes clinical and genetic associations between autoimmune disease and PD, impaired cellular and humoral immune responses in PD, imaging evidence of inflammatory cell activation and evidence of immune dysregulation in experimental models of PD. However, the mechanisms that link the immune system with PD remain unclear, and the temporal relationships of innate and adaptive immune responses with neurodegeneration are unknown. Despite these challenges, our current knowledge provides opportunities to develop immune-targeted therapeutic strategies for testing in PD, and clinical studies of some approaches are under way. In this Review, we provide an overview of the clinical observations, preclinical experiments and clinical studies that provide evidence for involvement of the immune system in PD and that help to define the nature of this association. We consider autoimmune mechanisms, central and peripheral inflammatory mechanisms and immunogenetic factors. We also discuss the use of this knowledge to develop immune-based therapeutic approaches, including immunotherapy that targets α-synuclein and the targeting of immune mediators such as inflammasomes. We also consider future research and clinical trials necessary to maximize the potential of targeting the immune system.
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http://dx.doi.org/10.1038/s41582-020-0344-4DOI Listing
June 2020

Lactobacilli spp.: real-time evaluation of biofilm growth.

BMC Microbiol 2020 03 24;20(1):64. Epub 2020 Mar 24.

Texas Tech University Health Sciences Center at the Permian Basin, 701 W. 5th Street, Odessa, TX, 79763, USA.

Background: Biofilm is a fundamental bacterial survival mode which proceeds through three main generalized phases: adhesion, maturation, and dispersion. Lactobacilli spp. (LB) are critical components of gut and reproductive health and are widely used probiotics. Evaluation of time-dependent mechanisms of biofilm formation is important for understanding of host-microbial interaction and development of therapeutic interventions. Time-dependent LB biofilm growth was studied in two systems: large biofilm output in continuous flow system (microfermenter (M), Institute Pasteur, France) and electrical impedance-based real time label-free cell analyzer (C) (xCELLigence, ACEA Bioscience Inc., San Diego, CA). L. plantarum biofilm growth in M system was video-recorded, followed by analyses using IMARIS software (Bitplane, Oxford Instrument Company, Concord, MA, USA). Additionally, whole genome expression and analyses of attached (A) and dispersed (D) biofilm phases at 24 and 48 h were performed.

Results: The dynamic of biofilm growth of L. plantarum was similar in both systems except for D phases. Comparison of the transcriptome of A and D phases revealed, that 121 transcripts differ between two phases at 24 h. and 35 transcripts - at 48 h. of M growth. The main pathways, down-regulated in A compared to D phases after 24 h. were transcriptional regulation, purine nucleotide biosynthesis, and L-aspartate biosynthesis, and the upregulated pathways were fatty acid and phospholipid metabolism as well as ABC transporters and purine nucleotide biosynthesis. Four LB species differed in the duration and amplitude of attachment phases, while growth phases were similar.

Conclusion: LB spp. biofilm growth and propagation area dynamic, time-dependent processes with species-specific and time specific characteristics. The dynamic of LB biofilm growth agrees with published pathophysiological data and points out that real time evaluation is an important tool in understanding growth of microbial communities.
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http://dx.doi.org/10.1186/s12866-020-01753-3DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7092459PMC
March 2020

Virtual Microscopy Is More Effective Than Conventional Microscopy for Teaching Cytology to Veterinary Students: A Randomized Controlled Trial.

J Vet Med Educ 2020 Jul 27;47(4):475-481. Epub 2020 Feb 27.

Director of the Academy for Teaching and Learning, College of Veterinary Medicine and Biomedical Sciences, Colorado State University.

Virtual microscopy (VM) using scanned slides and imaging software is increasingly used in medical curricula alongside instruction in conventional microscopy (CM). Limited reports suggest that VM is useful in the veterinary education setting, and generally well-received by students. Whether students can apply knowledge gained through VM to practical use is unknown. Our objective was to determine whether instruction using VM, compared to CM, is a successful method of training veterinary students for the application of cytology in practice (i.e., using light microscopes). Seventy-one veterinary students from Colorado State University who attended a voluntary 3-hour cytology workshop were randomized to receive the same instruction with either VM ( = 35) or CM ( = 36). We compared these students to a control group ( = 22) of students who did not attend a workshop. All students took a post-workshop assessment involving the interpretation of four cases on glass slides with CM, designed to simulate the use of cytology in general practice. Students also took an 18-question survey related to the effectiveness of the workshop, providing their opinions on cytology instruction in the curriculum and their learning preference (VM or CM). The mean assessment score of the VM group (14.18 points) was significantly higher than the control group (11.33 points, = .003), whereas the mean of the CM group (12.77 points) was not statistically significantly different from controls ( = .170). Not only is VM an effective method of teaching cytology to veterinary students that can be translated to a real-world case scenario, but it outperformed CM instruction in this study.
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http://dx.doi.org/10.3138/jvme.0318-029r1DOI Listing
July 2020

Comparison of in-sight and handheld navigation devices toward supporting industry 4.0 supply chains: First and last mile deliveries at the human level.

Appl Ergon 2020 Jan 21;82:102928. Epub 2019 Aug 21.

Design School, Loughborough University, UK.

Last (and First) mile deliveries are an increasingly important and costly component of supply chains especially those that require transport within city centres. With reduction in anticipated manufacturing and delivery timescales, logistics personnel are expected to identify the correct location (accurately) and supply the goods in appropriate condition (safe delivery). Moving towards more environmentally sustainable supply chains, the last/first mile of deliveries may be completed by a cyclist courier which could result in significant reductions in congestion and emissions in cities. In addition, the last metres of an increasing number of deliveries are completed on foot i.e. as a pedestrian. Although research into new technologies to support enhanced navigation capabilities is ongoing, the focus to date has been on technical implementations with limited studies addressing how information is perceived and actioned by a human courier. In the research reported in this paper a comparison study has been conducted with 24 participants evaluating two examples of state-of-the-art navigation aids to support accurate (right time and place) and safe (right condition) navigation. Participants completed 4 navigation tasks, 2 whilst cycling and 2 whilst walking. The navigation devices under investigation were a handheld display presenting a map and instructions and an in-sight monocular display presenting text and arrow instructions. Navigation was conducted in a real-world environment in which eye movements and device interaction were recorded using Tobii-Pro 2 eye tracking glasses. The results indicate that the handheld device provided better support for accurate navigation (right time and place), with longer but less frequent gaze interactions and higher perceived usability. The in-sight display supported improved situation awareness with a greater number of hazards acknowledged. The benefits and drawbacks of each device and use of visual navigation support tools are discussed.
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http://dx.doi.org/10.1016/j.apergo.2019.102928DOI Listing
January 2020

Actioning our understanding of respiratory compromise.

Can J Respir Ther 2019 15;55:28-29. Epub 2019 Feb 15.

Canadian Society of Respiratory Therapists, Saint John, NB, Canada.

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http://dx.doi.org/10.29390/cjrt-2018-026DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6591781PMC
February 2019

Proteomic analysis of urinary extracellular vesicles reveal biomarkers for neurologic disease.

EBioMedicine 2019 Jul 20;45:351-361. Epub 2019 Jun 20.

Center for Neurodegeneration and Neurotherapeutics, Duke University, Durham, NC, USA. Electronic address:

Background: Extracellular vesicles (EVs) harbor thousands of proteins that hold promise for biomarker development. Usually difficult to purify, EVs in urine are relatively easily obtained and have demonstrated efficacy for kidney disease prediction. Herein, we further characterize the proteome of urinary EVs to explore the potential for biomarkers unrelated to kidney dysfunction, focusing on Parkinson's disease (PD).

Methods: Using a quantitative mass spectrometry approach, we measured urinary EV proteins from a discovery cohort of 50 subjects. EVs in urine were classified into subgroups and EV proteins were ranked by abundance and variability over time. Enriched pathways and ontologies in stable EV proteins were identified and proteins that predict PD were further measured in a cohort of 108 subjects.

Findings: Hundreds of commonly expressed urinary EV proteins with stable expression over time were distinguished from proteins with high variability. Bioinformatic analyses reveal a striking enrichment of endolysosomal proteins linked to Parkinson's, Alzheimer's, and Huntington's disease. Tissue and biofluid enrichment analyses show broad representation of EVs from across the body without bias towards kidney or urine proteins. Among the proteins linked to neurological diseases, SNAP23 and calbindin were the most elevated in PD cases with 86% prediction success for disease diagnosis in the discovery cohort and 76% prediction success in the replication cohort.

Interpretation: Urinary EVs are an underutilized but highly accessible resource for biomarker discovery with particular promise for neurological diseases like PD.
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http://dx.doi.org/10.1016/j.ebiom.2019.06.021DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6642358PMC
July 2019

A Competency-Guided Veterinary Curriculum Review Process.

J Vet Med Educ 2020 Apr 13;47(2):137-147. Epub 2019 Jun 13.

Department of Clinical Sciences, Colorado State University Veterinary Teaching Hospital.

Competencies can guide outcomes assessment in veterinary medical education by providing a core set of specific abilities expected of new veterinary graduates. A competency-guided evaluation of Colorado State University's (CSU) equine veterinary curriculum was undertaken via an alumni survey. Published competencies for equine veterinary graduates were used to develop the survey, which was distributed to large animal alumni from CSU's Doctor of Veterinary Medicine program. The results of the survey indicated areas for improvement, specifically in equine business, surgery, dentistry, and radiology. The desire for more hands-on experiences in their training was repeatedly mentioned by alumni, with the largest discrepancies between didactic knowledge and hands-on skills in the areas of business and equine surgery. Alumni surveys allow graduates to voice their perceived levels of preparation by the veterinary program and should be used to inform curriculum revisions. It is proposed that the definition and utilization of competencies in each phase of a curricular review process (outcomes assessment, curriculum mapping, and curricular modifications), in addition to faculty experience and internal review, is warranted.
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http://dx.doi.org/10.3138/jvme.1217-183r1DOI Listing
April 2020

Caught in the act: LRRK2 in exosomes.

Biochem Soc Trans 2019 04 5;47(2):663-670. Epub 2019 Mar 5.

Duke Center for Neurodegeneration and Neurotherapeutics, Duke University, Durham, NC 27710, U.S.A.

Mutations in the (LRRK2) gene are a frequent genetic cause of late-onset Parkinson's disease (PD) and a target for therapeutic approaches. LRRK2 protein can influence vesicle trafficking events in the cytosol, with action both in endosomal and lysosomal pathways in different types of cells. A subset of late endosomes harbor intraluminal vesicles that can be secreted into the extracellular milieu. These extracellular vesicles, called exosomes, package LRRK2 protein for transport outside the cell into easily accessed biofluids. Both the cytoplasmic complement of LRRK2 as well as the exosome-associated fraction of protein appears regulated in part by interactions with 14-3-3 proteins. LRRK2 inside exosomes have disease-linked post-translational modifications and are relatively stable compared with unprotected proteins in the extracellular space or disrupted cytosolic compartments. Herein, we review the biology of exosome-associated LRRK2 and the potential for utility in diagnosis, prognosis, and theragnosis in PD and other LRRK2-linked diseases.
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http://dx.doi.org/10.1042/BST20180467DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6944476PMC
April 2019

The unlikely partnership between LRRK2 and α-synuclein in Parkinson's disease.

Eur J Neurosci 2019 02 24;49(3):339-363. Epub 2018 Oct 24.

Neurodegenerative Diseases Laboratory, UMR9199, CEA, CNRS, Université Paris Sud, Université Paris-Saclay, and MIRCen (Molecular Imaging Research Centre), Institut François Jacob, DRF, CEA, Fontenay-aux-Roses, France.

Our understanding of the mechanisms underlying Parkinson's disease, the once archetypical nongenetic neurogenerative disorder, has dramatically increased with the identification of α-synuclein and LRRK2 pathogenic mutations. While α-synuclein protein composes the aggregates that can spread through much of the brain in disease, LRRK2 encodes a multidomain dual-enzyme distinct from any other protein linked to neurodegeneration. In this review, we discuss emergent datasets from multiple model systems that suggest these unlikely partners do interact in important ways in disease, both within cells that express both LRRK2 and α-synuclein as well as through more indirect pathways that might involve neuroinflammation. Although the link between LRRK2 and disease can be understood in part through LRRK2 kinase activity (phosphotransferase activity), α-synuclein toxicity is multilayered and plausibly interacts with LRRK2 kinase activity in several ways. We discuss common protein interactors like 14-3-3s that may regulate α-synuclein and LRRK2 in disease. Finally, we examine cellular pathways and outcomes common to both mutant α-synuclein expression and LRRK2 activity and points of intersection. Understanding the interplay between these two unlikely partners in disease may provide new therapeutic avenues for PD.
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http://dx.doi.org/10.1111/ejn.14182DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6391223PMC
February 2019

Chemical kinetics in an atmospheric pressure helium plasma containing humidity.

Phys Chem Chem Phys 2018 Sep;20(37):24263-24286

York Plasma Institute, Department of Physics, University of York, Heslington, York YO10 5DD, UK.

Atmospheric pressure plasmas are sources of biologically active oxygen and nitrogen species, which makes them potentially suitable for the use as biomedical devices. Here, experiments and simulations are combined to investigate the formation of the key reactive oxygen species, atomic oxygen (O) and hydroxyl radicals (OH), in a radio-frequency driven atmospheric pressure plasma jet operated in humidified helium. Vacuum ultra-violet high-resolution Fourier-transform absorption spectroscopy and ultra-violet broad-band absorption spectroscopy are used to measure absolute densities of O and OH. These densities increase with increasing H2O content in the feed gas, and approach saturation values at higher admixtures on the order of 3 × 1014 cm-3 for OH and 3 × 1013 cm-3 for O. Experimental results are used to benchmark densities obtained from zero-dimensional plasma chemical kinetics simulations, which reveal the dominant formation pathways. At low humidity content, O is formed from OH+ by proton transfer to H2O, which also initiates the formation of large cluster ions. At higher humidity content, O is created by reactions between OH radicals, and lost by recombination with OH. OH is produced mainly from H2O+ by proton transfer to H2O and by electron impact dissociation of H2O. It is lost by reactions with other OH molecules to form either H2O + O or H2O2. Formation pathways change as a function of humidity content and position in the plasma channel. The understanding of the chemical kinetics of O and OH gained in this work will help in the development of plasma tailoring strategies to optimise their densities in applications.
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http://dx.doi.org/10.1039/c8cp02473aDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6161623PMC
September 2018

The G2019S mutation in LRRK2 imparts resiliency to kinase inhibition.

Exp Neurol 2018 11 24;309:1-13. Epub 2018 Jul 24.

Center for Neurodegeneration and Experimental Therapeutics, Department of Neurology, University of Alabama at Birmingham, Birmingham, AL 35233, USA. Electronic address:

The G2019S mutation in LRRK2 is one of the most common known genetic causes of neurodegeneration and Parkinson disease (PD). LRRK2 mutations are thought to enhance LRRK2 kinase activity. Efficacious small molecule LRRK2 kinase inhibitors with favorable drug properties have recently been developed for pre-clinical studies in rodent models, and inhibitors have advanced to safety trials in humans. Rats that express human G2019S-LRRK2 protein and G2019S-LRRK2 knock-in mice provide newly characterized models to better understand the ostensible target for inhibitors. Herein, we explore the relationships between LRRK2 kinase inhibition in the brain and the periphery to establish the link between LRRK2 kinase activity and protein stability, induction of lysosomal defects in kidney and lung, and how G2019S-LRRK2 expression impacts these phenotypes. Using a novel ultra-sensitive scalable assay based on protein capillary electrophoresis with LRRK2 kinase inhibitors included in-diet, G2019S-LRRK2 protein was resilient to inhibition compared to wild-type (WT)-LRRK2 protein, particularly in the brain. Whereas WT-LRRK2 kinase activity could be completed blocked without lowering LRRK2 protein levels, higher inhibitor concentrations were necessary to fully reduce G2019S-LRRK2 activity. G2019S-LRRK2 expression afforded robust protection from inhibitor-induced kidney lysosomal defects, suggesting a gain-of-function for the mutation in this phenotype. In rodents treated with inhibitors, parallel measurements of phospho-Rab10 revealed a poor correlation to phospho-LRRK2, likely due to cells that express Rab10 but poorly express LRRK2 in heterogenous tissues and cell isolates. In summary, our results highlight several challenges associated with the inhibition of the G2019S-LRRK2 kinase that might be considered in initial clinical efforts.
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http://dx.doi.org/10.1016/j.expneurol.2018.07.012DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7041630PMC
November 2018

Sensitivity and specificity of phospho-Ser129 α-synuclein monoclonal antibodies.

J Comp Neurol 2018 08;526(12):1978-1990

Department of Neurology, University of Alabama at Birmingham, Birmingham, Alabama.

α-Synuclein (α-syn) is an abundant presynaptic protein that is the primary constituent of inclusions that define Lewy body diseases (LBDs). In these inclusions, α-syn is phosphorylated at the serine-129 residue. Antibodies directed to this phosphorylation site are used to measure inclusion abundance and stage disease progression in preclinical models as well as in postmortem tissues in LBDs. While it is critical to reliably identify inclusions, phospho-specific antibodies often cross-react with nonspecific antigens. Four commercially available monoclonal antibodies, two from rabbits (clones EP1536Y and MJF-R13) and two from mice (81a and pSyn#64), have been the most widely used in detecting pS129-α-syn inclusions. Here, we systematically evaluated these antibodies in brain sections and protein lysates from rats and mice. All antibodies detected pS129-α-syn inclusions in the brain that were induced by preformed α-syn fibrils in wild-type rats and mice. Antibody titrations revealed that clones EP1536Y and 81a comparably labeled inclusions in both the perikarya and neuronal processes in contrast to clones MJF-R13 and pSyn#64 that incompletely labeled inclusions at various antibody concentrations. Except for EP1536Y, the clones produced nonspecific diffuse neuropil labeling in α-syn knockout mice as well as mice and rats injected with monomeric α-syn, with some nonspecific staining titrating with pS129-α-syn inclusions. By immunoblot, all the clones cross-reacted with proteins other than α-syn, warranting caution in interpretations of specificity. Clone EP1536Y uniquely and robustly detected endogenous pS129-α-syn in highly soluble protein fractions from the mouse brain. In summary, EP1536Y had the highest sensitivity and specificity for detecting pS129-α-syn.
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http://dx.doi.org/10.1002/cne.24468DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6031478PMC
August 2018

Sex Ed to Go: A Content Analysis of Comprehensive Sexual Education Apps.

Health Educ Behav 2018 08 28;45(4):581-590. Epub 2017 Dec 28.

1 University of New Mexico, Albuquerque, NM, USA.

Mobile applications ("apps") designed for sexual health education have the potential to reach teens and young adults that are hard to reach through traditional platforms; however, little is known about availability of these apps and their adherence to existing guidelines. Following a search on the two major app stores, data from 2,693 apps were analyzed. Only 697 (25%) addressed sexual health, and only 15 (1%) of apps met inclusion criteria for comprehensive programs and their content was further analyzed. The content of most of these apps narrowly focused on sexually transmitted infections and pregnancy prevention and lacked information on puberty, sexual identity, and personal safety. Theoretically grounded strategies including self-efficacy and modeling behavior to strengthen behavior change efforts were largely absent. Last, we identified significant shortcomings in the literate design of these apps, including limited use of interactive features, such as videos, quizzes, or games. These findings indicate that the potential of apps as sexual health promotion tools has not yet been fully realized. We outline recommendations for developing theory- and evidence-based sexual education apps and provide suggestions for health educators on how to select relevant apps when working for youth.
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http://dx.doi.org/10.1177/1090198117749259DOI Listing
August 2018

LRRK2 phosphorylates membrane-bound Rabs and is activated by GTP-bound Rab7L1 to promote recruitment to the trans-Golgi network.

Hum Mol Genet 2018 01;27(2):385-395

Department of Neurology, Center for Neurodegeneration and Experimental Therapeutics, University of Alabama at Birmingham, Birmingham, AL 35233 USA.

Human genetic studies implicate LRRK2 and RAB7L1 in susceptibility to Parkinson disease (PD). These two genes function in the same pathway, as knockout of Rab7L1 results in phenotypes similar to LRRK2 knockout, and studies in cells and model organisms demonstrate LRRK2 and Rab7L1 interact in the endolysosomal system. Recently, a subset of Rab proteins have been identified as LRRK2 kinase substrates. Herein, we find that Rab8, Rab10, and Rab7L1 must be membrane and GTP-bound for LRRK2 phosphorylation. LRRK2 mutations that cause PD including R1441C, Y1699C, and G2019S all increase LRRK2 phosphorylation of Rab7L1 four-fold over wild-type LRRK2 in cells, resulting in the phosphorylation of nearly one-third the available Rab7L1 protein in cells. In contrast, the most common pathogenic LRRK2 mutation, G2019S, does not upregulate LRRK2-mediated phosphorylation of Rab8 or Rab10. LRRK2 interaction with membrane and GTP-bound Rab7L1, but not Rab8 or Rab10, results in the activation of LRRK2 autophosphorylation at the serine 1292 position, required for LRRK2 toxicity. Further, Rab7L1 controls the proportion of LRRK2 that is membrane-associated, and LRRK2 mutations enhance Rab7L1-mediated recruitment of LRRK2 to the trans-Golgi network. Interaction studies with the Rab8 and Rab10 GTPase-activating protein TBC1D4/AS160 demonstrate that LRRK2 phosphorylation may block membrane and GTP-bound Rab protein interaction with effectors. These results suggest reciprocal regulation between LRRK2 and Rab protein substrates, where Rab7L1-mediated upregulation of LRRK2 kinase activity results in the stabilization of membrane and GTP-bound Rab proteins that may be unable to interact with Rab effector proteins.
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http://dx.doi.org/10.1093/hmg/ddx410DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5886198PMC
January 2018

Elevated LRRK2 autophosphorylation in brain-derived and peripheral exosomes in LRRK2 mutation carriers.

Acta Neuropathol Commun 2017 Nov 22;5(1):86. Epub 2017 Nov 22.

Department of Neurology, Center for Neurodegeneration and Experimental Therapeutics, University of Alabama at Birmingham, Birmingham, AL, 35233, USA.

Missense mutations in the leucine-rich repeat kinase 2 (LRRK2) gene can cause late-onset Parkinson disease (PD). LRRK2 mutations increase LRRK2 kinase activities that may increase levels of LRRK2 autophosphorylation at serine 1292 (pS1292) and neurotoxicity in model systems. pS1292-LRRK2 protein can be packaged into exosomes and measured in biobanked urine. Herein we provide evidence that pS1292-LRRK2 protein is robustly expressed in cerebral spinal fluid (CSF) exosomes. In a novel cohort of Norwegian subjects with and without the G2019S-LRRK2 mutation, with and without PD, we quantified levels of pS1292-LRRK2, total LRRK2, and other exosome proteins in urine from 132 subjects and in CSF from 82 subjects. CSF and urine were collected from the same morning clinic visit in 55 of the participants. We found that total LRRK2 protein concentration was similar in exosomes purified from either CSF or urine but the levels did not correlate. pS1292-LRRK2 levels were higher in urinary exosomes from male and female subjects with a LRRK2 mutation. Male LRRK2 mutation carriers without PD had intermediate pS1292-LRRK2 levels compared to male carriers with PD and controls. However, female LRRK2 mutation carriers without PD had the same pS1292-LRRK2 levels compared to female carriers with PD. pS1292-LRRK2 levels in CSF exosomes were near saturated in most subjects, ten-fold higher on average than pS1292-LRRK2 levels in urinary exosomes, irrespective of LRRK2 mutation status or PD diagnosis. These results provide insights into the effects of LRRK2 mutations in both the periphery and brain in a well-characterized clinical population and show that LRRK2 protein in brain exosomes may be much more active than in the periphery in most subjects.
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http://dx.doi.org/10.1186/s40478-017-0492-yDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5700679PMC
November 2017