Our laboratory is working with the human NTera2/D1 (NT2) cell line which have properties similar to progenitor cells in the CNS. These neural-like precursors cells can differentiate into all three major lineages - neurons, astrocytes, andoligodendrocytes. The pure neuronal population, called the hNT cells, possess characteristics of dopamine (DA) cells. In this dissertation, we performed various experiments to examine the neuronal and dopaminergic development of this cellline. We first cultured our hNT neurons with cells from the developingnigrostriatal (NS) pathway, the ventral mesencephalon and striatum, to determine their influence on survival, neuritic outgrowth, and DA phenotype. The survival ofhNT neurons was substantially greater when they were cultured with embryonicday (E) 18 cells, compared to monocultures or cocultures with either E14 orpostnatal day (P) 1 cells. The neuritic outgrowth of hNT neurons as assessed by the number of primary neurites per cell was increased when cultured with theareas of the brain from E14 and P1. The DA phenotype, as determined by the expression of the rate-limiting enzyme of DA synthesis was not increased in hNTneurons when they were cultured with primary rat cells from the NS pathway.Next we analyzed if the retinoic acid (RA)-treated hNT neurons and the NT2 precursor cells expressed three transcription factors required for development ofthe DA phenotype. We report that NT2 cells endogenously expressed Engrailed-1, Ptx3, and Nurr1 while RA treatment increased Nurr1 but down-regulated Engrailed-1 and Ptx3. Finally, lithium has been shown to stimulate neurogenesisin adult hippocampal precursors as well as influence the Wnt pathway known to be important for the induction of the DA phenotype.

<p>This thesis describes the utilization of the matrix-assisted laser desorption ionization mass spectrometry (MALDI MS) and electrospray ionization (ESI) MS techniques for analysis of complex brain tissue samples. </p><p>Direct molecular profiling of biological samples using MALDI MS is a powerful tool for identifying phenotypic markers. MALDI MS-profiling of proteins and peptides directly on brain tissue sections was used for the first time to study experimental models of Parkinson’s disease (PD). The mass spectrometer was used to map the peptide and protein expression directly on 12 µm tissue sections in mass-to-charge (m/z) values, providing the capability of mapping specific molecules of the original sample, that is, localization, intensity and m/z ratio. Several protein and peptide expression profile differences were found in the dopamine denervated brains when compared to the corresponding controls, for example, calmodulin, cytochrome c, cytochrome c oxidase, and the neuroimmunophilin protein FKBP-12. The increased expression of FKBP-12 from the profiling experiments was supported by mRNA expression analysis and two-dimensional gel electrophoresis separation analysis. Multiple genetic deficits have linked impaired ubiquitin-conjugation pathways to various forms of familiar PD. This study showed for the first time an increased level of unconjugated ubiquitin specifically in the dorsal striatum of the dopamine depleted PD brain. The strength of the MALDI MS-profiling technique is that a minimum of sample handling and manipulation is necessary pre-analysis. This ensures preservation of the spatial localization of the biomolecules in the tissue section.</p><p>Biological liquid samples often contain high amounts of salt that is non-compatible with the ESI MS technique. A nano-flow capillary liquid chromatography (nanoLC) system coupled on-line with ESI-MS was used to study the metabolism of the peptide LVV-hemorphin-7 in the brain and blood using in vivo microdialysis. The microdialysis technique provides capabilities for very precise sampling in specific brain regions. The combination of on-line desalting and pre-concentration by nanoLC with ESI MS is a powerful tool to detect minute concentration of metabolic fragments and endogenous biomolecules.</p><p>The utilization of mass spectrometry in neuroscience applications provides a uniquely advantageous tool for the analysis of complex biochemical events that underlie the pathological symptoms expressed in different disease states. Furthermore, the MALDI-MS profiling technique shows great potential for the future with regards to proteome analysis and drug discovery.</p>

This thesis describes the utilization of the matrix-assisted laser desorption ionization mass spectrometry (MALDI MS) and electrospray ionization (ESI) MS techniques for analysis of complex brain tissue samples. Direct molecular profiling of biological samples using MALDI MS is a powerful tool for identifying phenotypic markers. MALDI MS-profiling of proteins and peptides directly on brain tissue sections was used for the first time to study experimental models of Parkinson’s disease (PD). The mass spectrometer was used to map the peptide and protein expression directly on 12 µm tissue sections in mass-to-charge (m/z) values, providing the capability of mapping specific molecules of the original sample, that is, localization, intensity and m/z ratio. Several protein and peptide expression profile differences were found in the dopamine denervated brains when compared to the corresponding controls, for example, calmodulin, cytochrome c, cytochrome c oxidase, and the neuroimmunophilin protein FKBP-12. The increased expression of FKBP-12 from the profiling experiments was supported by mRNA expression analysis and two-dimensional gel electrophoresis separation analysis. Multiple genetic deficits have linked impaired ubiquitin-conjugation pathways to various forms of familiar PD. This study showed for the first time an increased level of unconjugated ubiquitin specifically in the dorsal striatum of the dopamine depleted PD brain. The strength of the MALDI MS-profiling technique is that a minimum of sample handling and manipulation is necessary pre-analysis. This ensures preservation of the spatial localization of the biomolecules in the tissue section. Biological liquid samples often contain high amounts of salt that is non-compatible with the ESI MS technique. A nano-flow capillary liquid chromatography (nanoLC) system coupled on-line with ESI-MS was used to study the metabolism of the peptide LVV-hemorphin-7 in the brain and blood using in vivo microdialysis. The microdialysis technique provides capabilities for very precise sampling in specific brain regions. The combination of on-line desalting and pre-concentration by nanoLC with ESI MS is a powerful tool to detect minute concentration of metabolic fragments and endogenous biomolecules. The utilization of mass spectrometry in neuroscience applications provides a uniquely advantageous tool for the analysis of complex biochemical events that underlie the pathological symptoms expressed in different disease states. Furthermore, the MALDI-MS profiling technique shows great potential for the future with regards to proteome analysis and drug discovery.

Background: Single photon emission computed tomography (SPECT) with dopamine transporter (DAT) and dopamine D2 receptor (D2R) ligands can visualise the integrity of the nigrostriatal dopamine system. Parkinson’s disease (PD) and the atypical parkinsonian diseases (APD), progressive supranuclear palsy (PSP) and multiple system atrophy (MSA), have similar symptoms and dopamine depletion, but differ in pharmacological response and prognosis. Clinical differentiation between PD and APD is often difficult in the early stages. The aims of the thesis were to evaluate the differential diagnostic and prognostic value of SPECT in early PD, MSA and PSP, to map the pattern of progression with dopamine SPECT, and map the pattern of dopamine SPECT in non-affected elderly volunteers with a prospective approach. Also, we evaluated the methodological aspects of dopamine SPECT with respect to image evaluation tools, reconstruction parameters and gamma cameras. Methods: 172 patients, included in an on-going clinical prospective study on idiopathic parkinsonism, participated in the SPECT study. Also, 31 age-matched healthy controls (HC) were followed within this study. SPECT was done with 123I-FP-Cit (DAT SPECT) and 123I-IBZM (D2R SPECT). Regions of interest (ROI) were used as a standard method for semi-quantitative image analysis. Results: SPECT uptake ratios from different gamma cameras could be equalised through correction equations derived from images of a brain-like phantom, provided that attenuation correction was applied. The ROI method had high reproducibility. SPECT uptake  in HC, measured with the ROI method and a volume based (VOI) method rendered similar trends, but gender and age differences in SPECT uptake were more marked with the VOI method, and less pronounced in DAT SPECT compared to D2R SPECT with both methods. The DAT SPECT uptake was significantly reduced in very early disease stage of PD and APD compared to HC. DATSPECT uptake was more reduced in PD with postural and gait disturbance (PIGD) compared to tremor-dominant PD. Decline in DAT SPECT uptake during the first year was more pronounced in PD and PSP compared to HC. D2R SPECT uptake overlapped between untreated PD and APD. After initiated treatment, the D2R SPECT uptake was significantly higher in MSA patients compared to PD, PSP and HC. Decline in D2R SPECT uptake during the first year was not significantly different between patients or compared to HC. Conclusions: 123I-FP-Cit SPECT is a valuable and sensitive method to detect early stage idiopathic parkinsonism. A different level of uptake between PIGD-PD compared to TD-PD indicates a prognostic potential. It is not possible to differ between PD, MSA and PSP in early stage with 123I-FP-Cit SPECT and no differential diagnostic value was found using 123I-IBZM SPECT in the early, untreated stage of PD, MSA and PSP. A different pattern of uptake of this ligand in MSA compared to PD and PSP during the first years of L-dopa treatment may, however, indicate a diagnostic value during the follow-up period.

Objective: In patients with Parkinson’s disease (PD), deep brainstimulation of the subthalamic nucleus (STN DBS) is well recognizedin improving limb function, but the outcome on swallowing functionhas rarely been studied. The aim of this work was to evaluate theeffect of STN DBS on pharyngeal swallowing function in patientswith PD using self-estimation and fiberoptic endoscopic evaluation ofswallowing. Methods: Eleven patients (aged 41–72, median 61 years)were evaluated preoperatively and at 6 and 12 months after STN DBSsurgery. All patients were evaluated with self-estimation on a visualanalogue scale, and eight of them with a fiberoptic endoscopicexamination with a predefined swallowing protocol includingRosenbek’s Penetration-Aspiration Scale, Secretion Severity Scale,preswallow spillage, pharyngeal residue, and pharyngeal clearance. Results: The self-assessments of swallowing function revealed asubjective improvement with STN DBS stimulation, whereas the datafrom the swallowing protocol did not show any significant effect ofthe STN DBS treatment itself. The prevalence of aspiration was notaffected by the surgery. Conclusions: The results show thatswallowing function was not negatively affected by STN DBS and therisk of aspiration did not increase. Self-estimation of swallowingfunction showed a subjective improvement due to stimulation / Speech, voice and swallowing outcomes after deep brain stimulation (DBS)

The disturbances and abnormalities occurring in the components of the Acetylcholine (ACh) neurocycle are considered one of the main features of cholinergic sicknesses like Parkinson’s and Alzheimer’s diseases. A fundamental understanding of the ACh neurocycle is therefore very critical in order to design drugs that keep the ACh concentrations in the normal physiological range. In this dissertation, a novel two-enzyme-two-compartment model is proposed in order to explore the bifurcation, dynamics, and chaotic characteristics of the ACh neurocycle. The model takes into consideration the physiological events of the choline uptake into the presynaptic neuron and the ACh release in the postsynaptic neuron. In order to approach more realistic behavior, two complete kinetic mechanisms for enzymatic processes pH-dependent are built: the first mechanism is for the hydrolysis reaction catalyzed by the acetylcholinesterase (AChE) and the other is for the synthesis reaction catalyzed by the cholineacetyltransferase (ChAT). The effects of hydrogen ion feed concentrations, AChE activity, ChAT activity, feed ACh concentrations, feed choline concentrations, and feed acetate concentrations as bifurcation parameters, on the system performance are studied. It was found that hydrogen ions play an important role, where they create potential differences through the plasma membranes. The concentrations of ACh, choline and acetate in compartments 1 and 2 are affected by the activity of AChE through a certain range of their concentrations, where the activity of AChE is inhibited completely after reaching certain values. A detailed bifurcation analysis over a wide range of parameters is carried out in order to uncover some important features of the system, such as hysteresis, multiplicity, Hopf bifurcation, period doubling, chaotic characteristics, and other complex dynamics. The effects of the feed choline concentrations and the feed acetate concentrations as bifurcation parameters are studied in this dissertation. It is found that the feed choline concentrations play an important role and have a direct effect on the ACh neurocycle through a certain important range of the parameters. However, the feed acetate concentrations have less effect. It is concluded from the results that the feed choline is a more important factor than the feed acetate in ACh processes. The effects of ChAT activity and the choline recycle ratio as bifurcation parameters, on the system performance are investigated. It was found that as the ChAT activity increases, ACh concentrations in compartments 1 and 2 increase continuously. The effect of the choline recycle ratio shows that choline reuptake plays a very critical role in the synthesis of ACh in compartment 1, where it supplies the choline as a substrate for the synthesis reaction by ChAT. The concentrations of ACh, choline and acetate in compartments 1 and 2 are affected by the choline recycle ratio through a certain range of the choline recycle ratio; then, they become constant as the choline recycle ratio increases further. It is concluded from our results that choline uptake is the rate limiting step in the ACh processes in both compartments in comparison to ChAT activity. Based on partial dissociation of the acetic acid in compartments 1, and 2 of the ACh cholinergic system, the two-parameter continuation technique has been applied to investigate the pH range to be closer to physiological ranges of pH values. In addition, static/dynamic solutions of the ACh cholinergic neurocycle system based on feed choline concentration as the main bifurcation parameter in both compartments have been investigated. The findings of the above studies are related to the real phenomena occurring in the neurons, like periodic stimulation of neural cells and non-regular functioning of ACh receptors. It was found that ACh, choline, acetate, and pH exist inside the physiological range associated with taking into consideration the partial dissociation of the acetic acid. The disturbances and irregularities (chaotic attractors) occurring in the ACh cholinergic system may be good indications of cholinergic diseases such as Alzheimer’s and Parkinson’s diseases. The results have been compared to the results of physiological experiments and other published models. As there is strong evidence that cholinergic brain diseases like Alzheimer’s disease and Parkinson’s disease are related to the concentration of ACh, the present findings are useful for uncovering some of the characteristics of these diseases and encouraging more physiological research.

Många närstående till en sjuk familjemedlem tar sig an rollen som vårdgivare vilket annars hade antagits av hälso- och sjukvårdspersonal. Nedskärningar inom den offentliga sektorn fortskrider och inte tillräckligt med fokus läggs på hur detta påverkar de närstående som vårdar en sjuk närstående i hemmet. Syftet med litteraturstudien var att beskriva upplevelsen och vad som påverkar upplevelsen av att vara vårdgivare i hemmet till en närstående med Parkinsons sjukdom. Studien utfördes som en litteraturstudie. I resultatet framkom fyra kategorier: Tid och relation, Stöd och information, Sjukdomsförlopp och Symtom. Resultatet visade att vårdgivare upplevde det svårt att ha en normalt fungerande vardag till följd av deras roll som vårdgivare. Vårdgivares upplevda börda ökade även i takt med den närståendes sjukdomsförlopp. En bra relation och mycket ömsesidighet mellan parterna minskade den upplevda bördan hos vårdgivarna samt ökade deras upplevda livskvalitet. Vårdgivare upplevde att stödgrupper minskar deras mentala börda genom att de fick möjlighet att dela med sig av sina erfarenheter till andra och att det hjälpte dem att förstå och själva handskas med sin situation. Vårdgivare upplevde att de inte fick tillräcklig information och stöd från hälso- och sjukvården. Vårdgivare ansåg att den fysiska funktionsnedsättningen var mer påtaglig vid direkt vård och gav mer fysisk påfrestning hos vårdgivaren än vad de psykiska symtomen gjorde. De psykiska symtomen hade större inverkan på vårdgivarens mentala hälsa och deras upplevda börda. Då antalet vårdgivare som vårdar en närstående i hemmet ökar är det av stor vikt att forskning genomförs och uppmärksammas. / Many people related to a sick person take on the role of caregiver, which would otherwise have been adopted by healthcare professionals. Cut downs in the public sector are progressing, and there is not enough focus on how this affects the persons who cares for a sick loved one at home. The purpose of this study was to describe the experience of being a caregiver and what factors affects the experience of a caregiver who cares for a loved one at home with Parkinson’s disease. The results revealed four categories: Time and relationship, Support and information, Disease duration and Symptoms. The results showed that caregivers find it difficult to have a normally functioning everyday life due to their role as caregivers. The caregivers perceived burden increases as the loved ones disease processes. A good relationship and a lot of mutuality between the parties reduce the perceived burden among caregivers, and increase their perceived quality of life. Caregivers feel that support groups reduce their mental workload by enabling them to share their experiences with others and that it helps them to understand and cope with their situation. Caregivers feel they do not receive adequate information and support from health care. The caregivers feel that the loved ones physical disability is more pronounced at the direct care and involves more physical stress than the mental symptoms makes. The psychological symptoms have a greater impact on caregivers’ mental health and their perceived burden. As the number of caregivers who care for a loved one at home increases it is of great importance to maintain research and attention to made research.

The disturbances and abnormalities occurring in the components of the Acetylcholine (ACh) neurocycle are considered one of the main features of cholinergic sicknesses like Parkinson’s and Alzheimer’s diseases. A fundamental understanding of the ACh neurocycle is therefore very critical in order to design drugs that keep the ACh concentrations in the normal physiological range. In this dissertation, a novel two-enzyme-two-compartment model is proposed in order to explore the bifurcation, dynamics, and chaotic characteristics of the ACh neurocycle. The model takes into consideration the physiological events of the choline uptake into the presynaptic neuron and the ACh release in the postsynaptic neuron. In order to approach more realistic behavior, two complete kinetic mechanisms for enzymatic processes pH-dependent are built: the first mechanism is for the hydrolysis reaction catalyzed by the acetylcholinesterase (AChE) and the other is for the synthesis reaction catalyzed by the cholineacetyltransferase (ChAT). The effects of hydrogen ion feed concentrations, AChE activity, ChAT activity, feed ACh concentrations, feed choline concentrations, and feed acetate concentrations as bifurcation parameters, on the system performance are studied. It was found that hydrogen ions play an important role, where they create potential differences through the plasma membranes. The concentrations of ACh, choline and acetate in compartments 1 and 2 are affected by the activity of AChE through a certain range of their concentrations, where the activity of AChE is inhibited completely after reaching certain values. A detailed bifurcation analysis over a wide range of parameters is carried out in order to uncover some important features of the system, such as hysteresis, multiplicity, Hopf bifurcation, period doubling, chaotic characteristics, and other complex dynamics. The effects of the feed choline concentrations and the feed acetate concentrations as bifurcation parameters are studied in this dissertation. It is found that the feed choline concentrations play an important role and have a direct effect on the ACh neurocycle through a certain important range of the parameters. However, the feed acetate concentrations have less effect. It is concluded from the results that the feed choline is a more important factor than the feed acetate in ACh processes. The effects of ChAT activity and the choline recycle ratio as bifurcation parameters, on the system performance are investigated. It was found that as the ChAT activity increases, ACh concentrations in compartments 1 and 2 increase continuously. The effect of the choline recycle ratio shows that choline reuptake plays a very critical role in the synthesis of ACh in compartment 1, where it supplies the choline as a substrate for the synthesis reaction by ChAT. The concentrations of ACh, choline and acetate in compartments 1 and 2 are affected by the choline recycle ratio through a certain range of the choline recycle ratio; then, they become constant as the choline recycle ratio increases further. It is concluded from our results that choline uptake is the rate limiting step in the ACh processes in both compartments in comparison to ChAT activity. Based on partial dissociation of the acetic acid in compartments 1, and 2 of the ACh cholinergic system, the two-parameter continuation technique has been applied to investigate the pH range to be closer to physiological ranges of pH values. In addition, static/dynamic solutions of the ACh cholinergic neurocycle system based on feed choline concentration as the main bifurcation parameter in both compartments have been investigated. The findings of the above studies are related to the real phenomena occurring in the neurons, like periodic stimulation of neural cells and non-regular functioning of ACh receptors. It was found that ACh, choline, acetate, and pH exist inside the physiological range associated with taking into consideration the partial dissociation of the acetic acid. The disturbances and irregularities (chaotic attractors) occurring in the ACh cholinergic system may be good indications of cholinergic diseases such as Alzheimer’s and Parkinson’s diseases. The results have been compared to the results of physiological experiments and other published models. As there is strong evidence that cholinergic brain diseases like Alzheimer’s disease and Parkinson’s disease are related to the concentration of ACh, the present findings are useful for uncovering some of the characteristics of these diseases and encouraging more physiological research.

The scope of this thesis is to process and analyze statistically Magnetic Resonance Images (MR-T1) from Parkinson’s disease patients in order to detect brain areas that exhibit brain change which is caused by the disease. Parkinson’s disease is an idiopathic disease which means that its cause is yet unknown. It is a chronic neurodegenerative disorder of the central nervous system which causes the progressive death of specific brain neurons that leads to motor impairments (tremor, bradykinesia, muscle rigidity) and non motor ones (cognitive, sleep, sensation disturbances). Magnetic Resonance Images (T1-weighted) were acquired from both Parkinson’s patients and healthy subjects (Controls) at intervals of 0 and 5 years. The data have undergone longitudinal (two-time-point), cross sectional (single-time-point) and statistical analysis with the use of FSL software library. Evidence of atrophy among Parkinson’s patients aroused, in brain areas near the ventricles and the middle temporal gyrus, after statistical analysis / Ο σκοπός αυτής της εργασίας είναι η επεξεργασία και στατιστική ανάλυση μαγνητηκών τομογραφιών (MR-T1) από ασθενείς με Πάρκινσον για την ανίχνευση περιοχών του εγκεφάλου που παρουσιάζουν μεταβολές που οφείλονται στην ασθένεια. Η ασθένεια Πάρκινσον είναι ιδιοπαθής, δηλαδή ασθένεια της οποίας η αιτία παραμένει ακόμη άγνωστη. Είναι μια χρόνια δυσλειτουργία λόγω εκφυλισμού των νευρώνων του κεντρικού νευρικού συστήματος η οποία προκαλεί τη σταδιακή νεκρωση συγκεκριμένης ομάδας εγκεφαλικών νευρώνων. Αυτή η νέκρωση οδηγεί σε κινητικές δυσλειτουργίες (τρέμουλο, βραδυκινησία, και μυϊκή δυσκαμψία και σε μή κινητικές όπως γνωστικές, διαταραχής ύπνου,διαταραχές αφής κ.α. Μαγνητικές τομογραφίες (τύπου Τ1) ασθενών και υγιών ελήφθησαν σε διαστήματα 0 και 5 ετών. Τα δεδομένα αναλύθηκαν με δυο μεθόδους (longitudinal και cross-sectional) και εν συνεχεία έγινε στατιστική επεξεργασία των αποτελεσμάτων. Έγινε χρήση της βιβλιοθήκης FSL Μετά από στατιστική ανάλυση προέκυψαν ενδείξεις ατροφίας στους ασθενείς με Πάρκινσον σε περιοχές του εγεκφάλου κοντά στις εγκεφαλικές κοιλίες (ventricles) και στη μέσο-κροταφική έλικα (middle temporal gyrus).

Background/Aim: Little is known about the clinical significance of phantosmia. The literature on phantosmia indicates that this symptom has a wide range of differential diagnoses. However, most cases of phantosmia remain of unknown origin. Our goal was to follow up patients with idiopathic phantosmia, with special regard to improvement rates and possible severe health conditions preceded by phantosmia of unknown origin. Methods: Forty-four patients with idiopathic phantosmia which had consulted our Ear-Nose-Throat Smell and Taste Clinic over the last 10 years were contacted by telephone and underwent a structured medical interview. Results: None of the patients had eveloped any severe health condition or Parkinson’s disease. More than 5 years after the occurrence of hantosmia, more than 50% of the patients experienced disappearance (31.8%) or improvement (25%). In the remaining cases, phantosmia did not change (38.7%) or became worse ( ! 5%). Conclusion: The main findings of the present study were that idiopathic phantosmia improves or disappears in almost two thirds of the patients after more than 5 years, and that idiopathic phantosmia seems to be more likely a harmless symptom rather than a reliable predictor of early Parkinson’s disease or other severe diseases. / Dieser Beitrag ist mit Zustimmung des Rechteinhabers aufgrund einer (DFG-geförderten) Allianz- bzw. Nationallizenz frei zugänglich.