A minimalistic co-culture platform for alpha-synuclein spreading in human dopaminergic neurons

Parkinson’s disease (PD) is the second major neurodegenerative disease and the most common movement disorder. Due to age being a critical risk factor, the rapid ageing of the world population further increases the prevalence of PD. So far no treatment is available and therapies mainly focus on motor symptoms by pharmacologically substituting striatal dopamine, caused by the loss of dopaminergic neurons in the substantia nigra. This neuronal loss and intracellular protein aggregates, termed Lewy bodies (LBs), are pathological characteristics of PD. With disease progression, a spread of LBs through the brain can be observed which mainly follows axonal projections. Understanding the mechanisms of this progressive spread could be central to discovering the underlying molecular pathogenesis of the disease. As LBs mainly consist of alpha-synuclein (-syn), a prion-like spreading of -syn was suggested and is now widely accepted as a component in the PD pathogenesis. New dopaminergic model systems to study the exact mechanisms underlying -syn spread are urgently needed. As PD is a human disease, in vitro models should be derived from humans. Lund human mesencephalic (LUHMES) cells are a suitable alternative to other, mostly non-human, dopaminergic cell lines. However, difficulties cultivating them in microfluidics devices has made them thus far inaccessible for co-cultivation studies in the field of PD spreading. In the first part of this thesis, a human dopaminergic cell model system for studying the spreading of -syn fibrils is presented. First, the well-characterized LUHMES cell line was tested for suitability of PD research on prion-like spreading, as no data is currently available on this matter. For the analysis, immunofluorescence light microscopy was employed. An extended period of differentiation aimed for a high degree of neuronal maturity and long neurites to facilitate the connectivity of spatially-separated cell populations. Seeding experiments with -syn fibrils revealed a weak toxicity against these assemblies, even at prolonged differentiation. Second, to study the transmission of -syn fibrils via neuronal projections, we developed a light microscopy-compatible microfluidic co-culturing device, to maintain two LUHMES cell populations in separate cell compartments for up to two weeks of differentiation. During this time, a neurite network is formed which connects the fluidically isolated cell growth compartments. The ability to cultivate cells with neurites and soma in an isolated environment enabled seeding and transmission experiments in anterograde and retrograde directions. In the second part of this thesis, implementation strategies of the microfluidic co-culturing chip for alternative analysis methods are discussed. Firstly, the accessibility of the cells in the co-culturing device using a single-cell lysis instrument is evaluated. The tool allows for targeted lysis of individual adherent cells. Preliminary tests point in a promising direction, while LUHMES single cell lysate was successfully transferred to different analysis techniques. However, direct access to the channels of the microfluidic co-culturing chip was problematic and needs further modifications. Secondly, an implementation of the microfluidic device aiming for co-cultivation of LUHMES cells on electron microscopy grids to study neurite architecture was pursued. Thereby, microfluidic devices harbor only cell soma, but neurites can grow onto an electron microscopy grid, as only they are thin enough to be visualized by cryo-electron microscopy. Proof-of-concept experiments demonstrate the direct visualization of LUHMES cell neurites in a near-native, frozen-hydrated state

Palliative care in Parkinson's disease : developing a needs assessment tool

Background: Parkinson’s disease (PD) is a common, life-limiting, neurodegenerative condition. Despite calls for improved access to palliative care, provision is lacking, due in part to poor understanding and recognition of palliative care needs. Where services exist, negotiating the chronic-palliative interface is challenging. Aims: 1) To establish the palliative needs of people living with PD. 2) To adapt and clinimetrically test a palliative assessment tool for PD. Method: A sequential mixed method design was adopted: Aim 1: Systematic review and synthesis of qualitative evidence and primary qualitative study (focus groups and semi-structured interviews). Combined findings used to adapt a palliative care assessment tool for Parkinson’s disease. Aim 2: Face and content validity tested by expert panel. Construct validity examined in 50 people with PD and their carers. Inter-rater reliability examined in broad range of clinicians using video consultations. Results: Aim 1 – Four lines of argument were identified: i) Information tension, ii) care tension, iii) interpersonal negotiations, iv) intra-personal negotiations. The response to diagnosis, carer vigilance and presence of disease milestones were highlighted. Aim 2 - Construct validity was good (tau B > 0.6) for two constructs, moderate (>0.4) for five and fair (>0.2) for the remainder. Reliability as measured by kappa was moderate (kappa >0.4) for four, fair (>0.2) for five and poor for four constructs. Two constructs with poor kappa are explained by extremely high percentage agreement. Conclusions: Adaptation of the NAT:Parkinson’s disease has been successful, with clinimetric properties supporting use by a range of clinicians. Future studies should examine utility within integrated services

Impact of a Community-Based Rock Steady Boxing Program for People with Parkinson\u27s Disease: A Pilot Study

Purpose/Hypothesis: High-intensity physical exercise has been shown to be beneficial in managing motor and nonmotor symptoms of Parkinson\u27s Disease (PD). Exercise may also have global effects on factors that influence brain health and cognition. Programs that incorporate goal-based motor skill learning have shown promise in being more effective than aerobic exercise alone. People with PD have a need for ongoing, continuous, community-based exercise programs that are engaging and accessible. The purpose of this study was to examine the effect of a community-based exercise program, Rock Steady Boxing, on improving quality of life and physical mobility skills in people with PD. Methods: Ten participants, five females and five males, mean age 69.6 years old (± 12 SD), clinically diagnosed with PD with a mean disease duration of 12.1 years (± 10 years) were recruited. Physical Therapist and Physical Therapy students collaborated with a local YMCA® to offer a Rock Steady boxing program (non-contact) to promote mobility, high intensity exercise, cognitive engagement, and transitional movements. Pre- and post-assessment at three months included gait speed, Parkinson\u27s Disease Questionnaire-39 (PDQ-39) for quality of life change, Mini BESTest to measure balance/mobility, Five Times Sit to Stand to measure strength, Four Square Step Test for agility, and the Six Minute Walk Test to measure endurance. Results: Five of the 10 participants reported an improvement in overall PDQ-39 scores. Of the 10 participants, 30% showed minimally clinically important change in cognition (MCID -1.8) and mobility (MCID -3.2), and 20% reported improvement in bodily discomfort (MCID -2.1). Post-test results of other measures were not statistically significant. Eight of the 10 participants attended the class, with a mean attendance of 6.7 visits (range 0-11), and indicated a plan to continue the exercise class. Statements from participants included: more confidence with walking and moving , more alert , and enjoy the social aspect. Three of the 10 also reported a reduction in falls, and one reported an increase in falls during the three-month timeframe. Conclusions: The outcomes of this pilot study show promise in improving quality of life and mobility in older adults with PD. Despite the progressive nature of PD, 30-50% of the participants in this study reported meaningful change in quality of life subscales while attending a three-month exercise program. More research is warranted to determine long-term benefits. Clinical Relevance: Community-based exercise programs tailored to people with PD appear beneficial in improving functional mobility and quality of life. Programs that incorporate a combination of high intensity exercise, skill-based training, cognitive engagement and social interaction are recommended. People with PD will benefit from consistent attendance at classes that are designed to be both physically and financially accessible, to minimize barriers and encourage long-term exercise participation

The Application and Development of Metabolomics Methodologies for the Profiling of Food and Cellular Toxicity

Metabolomics is a rapidly growing field of study. Its growth reflects advancements in technology and an improved understanding of the impact of the environment on metabolism. As a result, metabolomics is now commonly employed to investigate and characterize human and plant metabolism. The first chapter of this thesis provides an introduction to metabolomics and an overview of the protocols for sample preparation, data collection and statistical analysis. The second thesis chapter describes in explicit detail the step-by-step process of extracting and analyzing metabolites collected from mammalian cells, specifically brain tissue with a focus on Parkinson’s disease. The chapter highlights important factors to consider including experiment design, sample collection, and data processing. Chapters 3 and 4 include the application of metabolomics to evaluate how the metabolome responds to the environment. Chapter 3 focuses on the neuronal response to the xenobiotic arsenic. It demonstrates how astrocytes increase glutathione production through an up regulation of the citric acid cycle and glycolytic processes. Arsenic was also observed to decreases related metabolites including citrate and lactate. These metabolites are important intermediates to ATP production and illustrate the interconnection of metabolomic processes. Chapter 4 shows how metabolite profiles can be used to evaluate the impact of environmental conditions on wines. Metabolite profiles of Pinot Noir derived from the same scion clone (Pinot noir 667) and grown in different regions along the Pacific coast were compared. NMR and a differential sensing array were used to profile the chemical composition of the samples. We observed how environmental conditions resulted in different metabolite profiles in the various wine samples. This thesis aims to highlight the application of metabolomic to various biological studies in order to evaluate the impact of external stimuli. Advisor: Robert Power

Exploring the Role of Cognitive Reserve in Parkinson’s Disease Patient’s Responsiveness to Levodopa Treatment

This item is only available electronically.Parkinson’s disease (PD) is a progressive neurodegenerative disease characterised by dopamine loss. Dopamine medication as levodopa has been established to reduce PD motor symptoms and seemingly has erratic consequences for cognition. Cognitive reserve (CR), the mind’s resistance to brain damage, has been proposed to justify PD patients experiencing different impairments despite no significant neuropathological differences. Given the interaction between CR and levodopa state change is unexplored for PD, we sought to determine if CR predicts levodopa responsiveness and the role of other possible predictors. We tested six PD patient’s motor and fluid intelligence performance ON and OFF levodopa, to determine any variations in levodopa responsiveness. CR was measured with the comprehensive Cognitive Reserve Index questionnaire and a premorbid intelligence test. Motor and fluid intelligence performance was assessed with five precise measures of different abilities in ON and OFF levodopa states. Time limited data collection, meaning 80 PD patients were simulated from the six PD patients to meet regression conditions. We found CR did not significantly predict how PD patients responded to levodopa when controlling for age and levodopa equivalent dose (LED). However, LED was a significant predictor of change in motor and fluid intelligence performance, therefore acting as the most important contributing variable towards levodopa responsiveness. Age displayed a significant relationship with change in fluid intelligence performance. These findings suggest CR’s protective ability for PD was negligible when considering change in levodopa states and LED was the predominant levodopa responsiveness predictor. Implications for PD treating clinicians and patients are explored.Thesis (B.PsychSc(Hons)) -- University of Adelaide, School of Psychology, 202

PDapp - A mobile solution for continuous follow-up of Parkinson's disease patients

A doença de Parkinson é uma doença degenerativa que afeta principalmente neurónios produtores de dopamina, numa área específica do cérebro chamada substantia nigra. Os sintomas desenvolvem-se lentamente ao longo dos anos e a doença progride de uma forma muito variável de pessoa para pessoa. Estimulação Cerebral Profunda (DBS) é uma prática comum como tratamento nas fases mais tardias da doença. O sistema iHandU é um aparelho inovador que se pode equipar para medir a rigidez do pulso durante esta cirurgia. PDapp é uma aplicação móvel que procura trazer as funcionalidades da iHandU para fora da sala de cirurgia, de forma a ajudar os pacientes com um seguimento contínuo da sua doença e a sua progressão ao longo do tempo. Tem como objetivo criar uma ligação entre os pacientes e os seus médicos, fora do ambiente hospitalar. A PDapp fornece aos seus pacientes ferramentas para gerir a sua doença com medicação, testes e a possibilidade de informar os seus médicos de acontecimentos importantes relacionados à doença. Os médicos terão acesso a um dashboard especializado onde poderão analisar as informações dos seus doentes, o seu historial e acontecimentos recentes. O sistema será composto de uma aplicação móvel, uma plataforma web para o dashboard e uma base de dados na cloud. No final, teremos um sistema capaz de integrar a aplicação móvel dos pacientes, o dashboard dos médicos e a iHandU para fazer testes. A aplicação móvel terá de ser interativa e facilmente usável por pacientes com Parkinson's. O dashboard dos médicos será focado em apresentar a informação mais essencial de uma forma direta e costumizável, para que os médicos possam tomar melhores decisões. Um sistema como a PDapp pode ter grande impacto nas vidas de pacientes com Parkinson's, ajudando a gerir a doença e a liga-los aos seus médicos nos momentos importantes.Parkinson's disease is a neurodegenerative disorder that affects dopamine-producing ("dopaminergic") neurons predominately in a specific area of the brain called substantia nigra. Symptoms develop slowly over the years, and the disease's progression is often different from person to person. Deep Brain Stimulation is a common practice in the later stages of the disease as treatment. The iHandU system is a novel, comfortable, and wireless device, designed to classify the wrist rigidity during this practice. PDapp is a mobile app solution to bring the iHandU's capabilities out of the surgery room and help patients with a continuous follow up of their disease's progression through time. It strives to connect the patients and their doctors outside of the clinical environment. It provides patients with a high-quality mobile application for managing their disease with medication and tests, as well as keep their doctors informed of important events related to the disease. On the other hand, the doctors have access to a specialised dashboard to get an overview of all their patients' history and their most recent events. The system required will be composed of a mobile application, a web platform to host the dashboard, and a cloud-based database to persist all the data necessary. By the end of this project, we should have a system capable of successfully integrating a mobile application for the patients, a dashboard for the doctor, and the iHandU for performing tests. The mobile app should be highly usable and interactive for users with Parkinson's disease, and the dashboard should be focused and clean to provide the doctors with the best overview of their patients and current situations. A system like PDapp can have a significant impact on the lives of these patients by easing their access to relevant tools and connecting them to their doctors whenever needed

DNP Final Report: MEDICATION ADHERENCE: EVIDENCE-BASED INTERVENTIONS FOR PARKINSON\u27S DISEASE

Parkinson\u27s disease (PD) is a challenging, progressive neurodegenerative process projected to affect greater than one million patients in the United States over the next decade. Pharmacological interventions are the hallmark of treatment and are used for symptom management; there is no cure for this lifelong disease. Once-daily preparations are typically inadequate as a sole treatment or contraindicated related to comorbid or drug-induced psychiatric symptoms or behaviors. In addition, multiple medications and a variety of dosing times have an increasingly negative effect on medication adherence. The literature contains many resources to measure and understand what medication non-adherence is, who is at risk, and some novel ways to improve the outcomes. Being a chronic disease, PD follows similar consequences caused by inconsistent medication self-management. The scales imported from hypertension and kidney transplant patient education make it essential to know that no one scale will do it all. Comparatively, intervention strategies were also imported and seen throughout the literature, including time-tested patient education, newer ideas using technology like electronic pill dispensers, smartphone timers or reminders, simplified doses, telephone call follow-up, and support groups. The care of PD patients has come a long way. However, we must implement more effective interventions until the evidence shows higher medication adherence. This project aimed to illustrate how to recover PD patients’ quality of life through symptom management using evidenced-based interventions to improve medication non-adherence