Indiana University-Purdue University Indianapolis (IUPUI) / Parkinson's disease (PD) is a progressive neurodegenerative disorder that affects over 10 million people. Treatments for PD are limited to symptom mitigation with no means of stopping or slowing disease progression. Mutations within the protein leucine- rich repeat kinase 2 (LRRK2) are the most common cause of familial PD and are indistinguishable from the more common sporadic cases. Identifying molecules capable of modulating LRRK2 GTPase activity may serve as the foundation for future development of novel PD therapeutics. We recently discovered that the G-domain (ROC) of LRRK2 is capable of transitioning between monomer and dimer form in solution upon GTP/GDP binding. R1441C/G/H pathogenic mutations were demonstrated to alter this dynamic shifting toward a monomeric ROC conformation while decreasing GTPase activity. Using our ROC dimeric crystal structure, we strategically introduced disulfide bonds to generate locked monomer and locked dimer states. Monomeric ROC was shown to increase GTPase activity while the dimeric form decreased activity. Solvent mapping performed using the dimeric ROC crystal structure and a homology model of the ROC monomer revealed a binding hotspot at the ROC dimeric interface and adjacent to the R1441 residue in the monomeric model. In this study our goal was to identify more compounds capable of influencing GTPase activity. We performed high throughput screening of ROC against two compound libraries (LOPAC1280 and ChemBridge 50K) in a GTP binding assay. Twenty-three hits were identified and four compounds were further investigated in dose-response experiments. 3,4-Methylenedioxy-beta nitrostyrene (MNS) was demonstrated to decrease GTP binding and inhibit GTPase activity (IC50=23.92μM) while the compound N-phenylanthranilic acid increased GTP binding (EC50=4.969μM) and decreased GTPase activity. Identification of these compounds is the first step in the development of a novel PD therapeutic targeting the G-domain of LRRK2.

Summary: Movement disorders are caused by impairment of the basal ganglia and extrapyramidal connections. The most common is Parkinson's disease (PD), characterised by hypokinesia together with resting tremor and / or rigidity, which may be influenced by dopaminergic therapy or invasive methods, including deep brain stimulation (DBS). Gait and balance disorders are part of the Parkinson's syndrome, progress during the disease course and limit daily activities, quality of life, may lead to falls and contribute to higher mortality of the patients in the late stages. Therapy is difficult. Gait disorders and parkinsonism may occur also in other diseases, including intoxications which may also lead to basal ganglia impairment. The theoretical part include the physiology of gait, gait disorders and examination. The section devoted to movement disorders focuses on Parkinson's disease and methanol intoxication. The practical part consists of 3 studies evaluating gait disorder in patients with advanced PD and possibilities of the treatment, the 4th study documents gait disorder in survivors after mass methanol poisoning with outbreak between 2012 and 2014 in the Czech Republic. The result confirm the effect of DBS in step length prolongation and gait speed increment in patients with PD. Compensation strategies using...

Gait analysis allows clinicians and researchers to quantitatively characterize the kinematics and kinetics of human movement. Devices that quantify gait can be either portable, such as instrumented shoes, or non-portable, such as motion capture systems and instrumented walkways. There is a tradeoff between these two classes of systems in terms of portability and accuracy. However, recent computer advances allow for the collection of meaningful data outside of the clinical setting. In this work, we present the DeepSole system combined with the different neural network models. This system is a fully capable to characterize the gait of the individuals and provide vibratory feedback to the wearer. Thanks to the flexible construction and its wireless capabilities, it can be comfortably worn by wide arrange of people, both able-bodied and people with pathologies that affect their gait. It can be used for characterization, training, and as an abstract sensor to measure human gait in real-time. Three neural network models were designed and implemented to map the sensors embedded in the DeepSole system to gait characteristics and events. The first one is a recurrent neural network that classifies the gait into the correct gait phase of the wearer. This model was validated with data from healthy young adults and children with Cerebral Palsy. Furthermore, this model was implemented in real-time to provide vibratory feedback to healthy young adults to create temporal asymmetry on the dominant side during regular walking. During the experiment, the subjects who walked had an increased stance time on both sides, but the dominant side was affected more. The second model is encoder-decoder recurrent neural network that maps the sensors into current gait cycle percentage. This model is useful to provide continuous feedback that is synchronized to the gait. This model was implemented in real-time to provide vibratory feedback to six muscle groups used during regular walking. The effects of the vibration were analyzed. It was found that depending on the feedback, the subjects changed their spatial and temporal gait parameters. The third model uses all the sensors in the instrumented footwear to identify a motor phenomenon called freezing of gait in patients with Parkinson’s Disease. This phenomenon is characterized by transient periods, usually lasting for several seconds, in which attempted ambulation is halted. The model has better performance than the state-of-the-art and does not require any pre-processing. The DeepSole system when used in conjunction with the presented models is able to characterize and provide feedback in a wide range of scenarios. The system is portable, comfortable, and can accommodate a wide range of populations who can benefit from this wearable technology.

Parkinson’s disease is a common neurological disease which will progressively damagedopaminergic neurons in the brain. Later stages of the disease will result in death of theneurons. The diagnosis is often made with respect to the motor symptoms, which includetremors, bradykinesia, and rigidity. In addition to motor symptoms, non-motor symptomsappear in many patients, such as cognitive changes and mood disorders. One method used totreat Parkinson’s disease is deep brain stimulation, where electric pulses are emitted to aspecific brain area. A common target is the subthalamic nucleus, which is part of the basalganglia. By using deep brain stimulation, the dose of other medications for Parkinson’sdisease can be lowered. However, the mechanisms of deep brain stimulation are not yetentirely known, and there have been many reports of adverse side-effects caused by thismethod, including depression and other types of mood changes. Even so, information of apossible sex distribution of these side-effects is still limited. Here, a qualitative essay wasmade where 16 articles describing reported side-effects in men and women were compared. Inaddition, unpublished data from optogenetic studies on male and female mice were analysedin order to examine putative sex-differences upon experimental brain stimulation strategies.The results from the optogenetics results did not show any statistically significant sexdifferences.In contrast, by comparing the selected articles in which results of deep brainstimulation treatment in patients were reported, some differences were found. First, it seemsthat women report more depressive-like symptoms than men. Second, while men also reportdepressions, they also report more aggressive behaviour upon the treatment. A preliminaryconclusion of this essay is therefore that certain sex-differences can be observed among theadverse side-effects reported upon deep brain stimulation in Parkinson´s disease. However,since the studied material was limited, more research is required to make firmer conclusions.

Parkinson’s Disease is a degenerative neurological disease where dopaminergic neurons in the basal ganglia degenerate and die. This leads to dopamine deficiency in the basal ganglia, affecting both motor and non-motor systems, giving both motor and non-motor symptoms. Common non-motor symptoms in Parkinson’s disease are depression, anxiety and apathy. Exercise have shown to be effective in decreasing depression in otherwise healthy people. The aim of this degree project was to examine if exercise can act as a treatment against impaired mood for people with Parkinson's disease. Nine trials investigating the effect of different types of training with different duration and different exercise intervals were selected and studied. By comparing the results obtained in these trials, it was found that in seven of the nine studies, exercise have had a positive effect on depression. Fewer studies had examined anxiety and apathy, and the effect of exercise was not as clear. Anxiety decreased in two of three studies and apathy decreased in one of three. The studied material is limited but the conclusion that can be drawn from this study, is that exercise is a safe way to, without negative side effects, treat and/or counteract mood disorders including depressive symptoms in Parkinson’s disease, mild to moderate stage.

Because of conflicting reports of the actions of the antiparkinsonian agent L-prolyl-L-leucyl-glycine amide (PLG, MIF-I) on the turnover of Striatal dopamine (DA), this process was reinvestigated. In the present series of studies, it was found that neither our MIF-I (200 ng ICV) nor the MIF-I used by Versteeg et al. [25]was effective in altering the rate of decline of endogenous DA in the caudate nucleus of rats pretreated with α-methyl-p-tyrosine (300 mg/kg IP). In addition, our MIF-I (1 mg/kg IP) did not change endogenous dihydroxyphenylacetic acid (DOPAC) or homovanillic acid (HVA) in rat striatum. These studies indicate that MIF-I does not alter the turnover rate of DA in nigrostriatal neurons. It is possible that MIF-I or some substance released by MIF-I acts at a posfsynaptic receptor site.

The effect of chronic treatment with vitamin E (VE) on acute l-methyl-4-phenyl-l,2,3,6-tetrahydropyridine (MPTP)-induced neurotoxicity, as assessed by striatal dopamine (DA) depletion, was studied. Male C57B1/6J mice were fed VE (48 mg kg-1 per day, intragastric) for 4, 8, or 12 weeks prior to administration of MPTP (20 mg kg-1, i.p. X 3, 2 h intervals) or its diluent. Brain VE concentration was increased by exogenous supplements for 12 weeks. Striatal DA content was reduced by 85% to 90% after MPTP in control and VE-treated mice. Mice with elevated cerebral VE were not protected from MPTP toxicity, with DA content as an indicator. In conclusion, these findings indicate that moderate elevation of brain VE is not adequate for protecting DA-containing neurons against the toxic actions of a high dose of MPTP.

Parkinson's disease is a progressive neurodegenerative movement disorder, affecting mainly the elderly. One of the most important hallmarks of Parkinson's disease is the loss of neuronal cell bodies containing neuromelanin in the substantia nigra zona compacta, and subsequently, loss of dopamine terminals in basal ganglia nuclei of the brain. The discovery by Hornykiewicz and co-workers that levodopa could successfully treat Parkinson's disease in humans was one of the most important events of medicine in the 20th century. Since loss of nigrostriatal dopaminergic function is the basic underlying pathophysiology of this disease, drugs that enhance dopaminergic function in the striatum, including the exogenous precursor levodopa, remain the most effective symptomatic agents in the treatment of Parkinson's disease. However, there are some areas of controversy about levodopa-evoked motor complications (dyskinesias, on-off phenomena) as well as neuroprotective or neurotoxic activity of this drug, etc. In this article the authors try to clarify the molecular mechanisms involved in levodopa action, such as volume transmission - a crucial process for successful levodopa therapy, evidence that serotoninergic neurons may accumulate levodopa and convert it into dopamine as well as some aspects of neuroprotective action of levoda.

L-Dihydroxyphenylalanine (L-DOPA), the anti-parkinsonian drug affording the greatest symptomatic relief of parkinsonian symptoms, is still misunderstood in terms of its neurotoxic potential and the mechanism by which generated dopamine (DA) is able to exert an effect despite the absence of DA innervation of target sites in basal ganglia. This review summaries important aspects and new developments on these themes. On the basis of L-DOPA therapy in animal models of Parkinson's disease, it appears that L-DOPA is actually neuroprotective, not neurotoxic, as indicated by L-DOPAs reducing striatal tissue content of the reactive oxygen species, hydroxyl radical (HO•), and by leaving unaltered the extraneuronal in vivo microdialysate level of HO•. In addition, the potential beneficial anti-parkinsonian effect of L-DOPA is actually increased because of the fact that the basal ganglia are largely DA-denervated. That is, from in vivo microdialysis studies it can be clearly demonstrated that extraneuronal in vivo microdialysate DA levels are actually higher in the DA-denervated vs. the intact striatum of rats - owing to the absence of DA transporter (i.e., uptake sites) on the absent DA nerve terminal fibers in parkinsonian brain. In essence, there are fewer pumps removing DA from the extraneuronal pool. Finally, the undesired motor dyskinesias that commonly accompany long-term L-DOPA therapy, can be viewed as an outcome of L-DOPAs sensitizing DA receptors (D1-D5), an effect easily replicated by repeated DA agonist treatments (especially agonist of the D 2 class) in animals, even if the brain is not DA-denervated. The newest findings demonstrate that L-DOPA induces BDNF release from corticostriatal fibers, which in-turn enhances the expression of D3 receptors; and that this effect is associated with motor dyskinesias (and it is blocked by D3 antagonists). The recent evidence on mechanisms and effects of L-DOPA increases our understanding of this benefical anti-parkinsonian drug, and can lead to improvements in L-DOPA effects while providing avenues for reducing or eliminating L-DOPAs deleterious effects.

Nucleotide coenzymes dot the map of metabolic pathways providing energy to drive the reactions of the pathway and play an important role in regulating and controlling energy metabolism through their shared potential energy, which is widely unobserved due to the paradox that the energy in the coenzyme pools cannot be determined from the concentration of the coenzyme couples. The potential energy of the nucleotide couples in the mitochondria or the cytoplasm is expressed in the enzyme reactions in which they take part. The energy in these couples, [NAD+]/[NADH], [NADP+]/[NADPH], [acetyl CoA]/[CoA], and [ATP]/[ADP]x[Pi], regulates energy metabolism. The energy contained in the couples can be altered by suppling energy equivalents in the form of ketones, such as, D-β-hydroxybutyrate to overcome insulin resistance, to restore antioxidants capacity, to form potential treatments for Alzheimer's and Parkinson's diseases, to enhance life span, and to increase physiological performance. © 2019 IUBMB Life, 71(5):565–579, 2019.