Cathepsin B p.Gly284Val Variant in Parkinson's Disease Pathogenesis.

Parkinson's disease (PD) is generally considered a sporadic disorder, but a strong genetic background is often found. The aim of this study was to identify the underlying genetic cause of PD in two affected siblings and to subsequently assess the role of mutations in Cathepsin B (CTSB) in susceptibility to PD. A typical PD family was identified and whole-exome sequencing was performed in two affected siblings. Variants of interest were validated using Sanger sequencing. CTSB p.Gly284Val was genotyped in 2077 PD patients and 615 unrelated healthy controls from the Czech Republic, Ireland, Poland, Ukraine, and the USA. The gene burden analysis was conducted for the CTSB gene in an additional 769 PD probands from Mayo Clinic Florida familial PD cohort. CTSB expression and activity in patient-derived fibroblasts and controls were evaluated by qRT-PCR, western blot, immunocytochemistry, and enzymatic assay. The CTSB p.Gly284Val candidate variant was only identified in affected family members. Functional analysis of CTSB patient-derived fibroblasts under basal conditions did not reveal overt changes in endogenous expression, subcellular localization, or enzymatic activity in the heterozygous carrier of the CTSB variant. The identification of the CTSB p.Gly284Val may support the hypothesis that the CTSB locus harbors variants with differing penetrance that can determine the disease risk.

Validation of the Polish version of the Unified Dyskinesia Rating Scale (UDysRS).

BACKGROUND: In 2008, the Movement Disorders Society published the Unified Dyskinesia Rating Scale (UDysRS). This has become the established tool for assessing the severity and disability associated with dyskinesia in patients with Parkinson's Disease (PD). We translated and validated the Polish version of the UDysRS, explored its dimensionality, and compared it to the Spanish version, which is the Reference Standard for UDysRS translations. MATERIAL AND METHODS: The UDysRS was translated into Polish by a team led by JS and GO. The back-translation, completed by colleagues fluent in both Polish and English who were not involved in the original translation, was reviewed and approved by the Executive Committee of the MDS Rating Scales Programme. Then the translated version of the UDysRS underwent cognitive pretesting, and the translation was modified based on the results. The approved version was considered to be the Official Working Document of the Polish UDysRS and was tested on 250 Polish PD patients recruited at movement disorder centres. Data was compared to the Reference Standard used for validating UDysRS translations. RESULTS: The overall factor structure of the Polish version was consistent with that of the Reference Standard version, as evidenced by the high Confirmatory Fit Index score (CFI = 0.98). The Polish UDysRS was thus confirmed to share a common factor structure with the Reference Standard. CONCLUSIONS: The Official Polish UDysRS translation is recommended for use in clinical and research settings. Worldwide use of uniform rating measures offers a common ground to study similarities and differences in disease manifestations and progression across cultures.

Validation of the Polish version of the Movement Disorder Society-Unified Parkinson's Disease Rating Scale (MDS-UPDRS).

BACKGROUND: In 2008, the Movement Disorders Society (MDS) published a new Unified Parkinson's Disease Rating Scale (MDS-UPDRS) as the official benchmark scale for Parkinson's Disease (PD). We have translated and validated the Polish version of the MDS-UPDRS, explored its dimensionality, and compared it to the original English one. METHODS: The MDS-UPDRS was translated into Polish by a team of Polish investigators led by JS and GO. The back-translation was completed by colleagues fluent in both languages (Polish and English) who were not involved in the original translation, and was reviewed by members of the MDS Rating Scales Programme. Then the translated version of the MDS-UPDRS underwent cognitive pretesting, and the translation was modified based on the results. The final translation was approved as the Official Working Document of the MDS-UPDRS Polish version, and was tested on 355 Polish PD patients recruited at movement disorders centres all over Poland (at Katowice, Gdansk, Lódz, Warsaw, Wroclaw, and Kraków). Confirmatory and explanatory factor analyses were applied to determine whether the factor structure of the English version could be confirmed in the Polish version. RESULTS: The Polish version of the MDS-UPDRS showed satisfactory clinimetric properties. The internal consistency of the Polish version was satisfactory. In the confirmatory factor analysis, all four parts had greater than 0.90 comparative fit index (CFI) compared to the original English MDS-UPDRS. Explanatory factor analysis suggested that the Polish version differed from the English version only within an acceptable range. CONCLUSIONS AND CLINICAL IMPLICATIONS: The Polish version of the MDS-UPDRS meets the requirements to be designated as the Official Polish Version of the MDS-UPDRS, and is available on the MDS web page. We strongly recommend using the MDS-UPDRS instead of the UPDRS for research purposes and in everyday clinical practice.

[Evaluation of heart rate and blood pressure variability in Parkinson's disease patients after bilateral subthalamic deep brain stimulation]. / Ocena zmiennosci rytmu serca i cisnienia tetniczego u osób z choroba Parkinsona po obustronnej glebokiej stymulacji jadra niskowzgórzowego.

INTRODUCTION: Autonomic dysfunctions are the most common non-motor symptoms of Parkinson's disease (PD) and often precede the motor symptoms of the disease. Autonomic dysfunction may be a dominant symptom of the advanced stages of PD as well as a major cause of patient disability. Despite the wide use of neurostimulation in clinical practice, the effect of deep brain stimulation of subthalamic nucleus (STN DBS) on autonomic symptoms of PD still remains only partially understood. The aim of the study is evaluation of heart rate variability (HRV) and blood pressure variability (BPV) in patients with PD before STN DBS and following bilateral STN DBS. MATERIAL AND METHODS: The study included 25 subjects aged between 31 and 71 years, diagnosed with the idiopathic PD and selected for treatment with STN DBS. All the patients were in advanced stages of PD, disease duration ranged from 5 to 22 years. The patients enrolled into this study underwent STN DBS. Neurological examination including assessment of the severity of parkinsonism according to UPDRS scale, a psychological examination and an electrophysiological examination of autonomic disturbances based on heart rate and blood pressure variability were conducted on all patients two weeks before and three months after STN DBS. RESULTS: After STN DBS an improvement in terms of the analyzed parts of the UPDRS has been shown. The improvement of motor disorders assessed by III part UPDRS during the "off" medication/stimulation "on" was 67.8%. Orthostatic hypotension before the STN DBS procedure was observed in 56% of patients and after STN DBS in 53% of them. Before STN DBS the imbalance of the sympathetic--parasympathetic components with the predominance of the sympathetic based on HRV parameters--the ratio LF/HF-RRI (2.5) and a higher rate of LFnu (61.3%) than HFnu (38.6%) has been shown. Three months post STN DBS an increase parameters of spectral analysis of HRV in the low frequency LF-RRI, and high-frequency HF-RRI and the total power spectrum PSD-RRI was observed. After STN DBS an increase of parameters of spectral analysis of systolic BPV, very low frequency VLF-sBP, low frequency LF-sBP and total power spectrum PSD-sBP was noted. CONCLUSIONS: Results of the study suggest that STN DBS is an effective treatment method of both motor symptoms and autonomic dysfunctions. The disturbances of HRV and BPV before and after STN DBS indicate the increase of autonomic system activity with sympathetic dominance.

VPS35 mutations in Parkinson disease.

The identification of genetic causes for Mendelian disorders has been based on the collection of multi-incident families, linkage analysis, and sequencing of genes in candidate intervals. This study describes the application of next-generation sequencing technologies to a Swiss kindred presenting with autosomal-dominant, late-onset Parkinson disease (PD). The family has tremor-predominant dopa-responsive parkinsonism with a mean onset of 50.6 ± 7.3 years. Exome analysis suggests that an aspartic-acid-to-asparagine mutation within vacuolar protein sorting 35 (VPS35 c.1858G>A; p.Asp620Asn) is the genetic determinant of disease. VPS35 is a central component of the retromer cargo-recognition complex, is critical for endosome-trans-golgi trafficking and membrane-protein recycling, and is evolutionarily highly conserved. VPS35 c.1858G>A was found in all affected members of the Swiss kindred and in three more families and one patient with sporadic PD, but it was not observed in 3,309 controls. Further sequencing of familial affected probands revealed only one other missense variant, VPS35 c.946C>T; (p.Pro316Ser), in a pedigree with one unaffected and two affected carriers, and thus the pathogenicity of this mutation remains uncertain. Retromer-mediated sorting and transport is best characterized for acid hydrolase receptors. However, the complex has many types of cargo and is involved in a diverse array of biologic pathways from developmental Wnt signaling to lysosome biogenesis. Our study implicates disruption of VPS35 and retromer-mediated trans-membrane protein sorting, rescue, and recycling in the neurodegenerative process leading to PD.

Translation initiator EIF4G1 mutations in familial Parkinson disease.

Genome-wide analysis of a multi-incident family with autosomal-dominant parkinsonism has implicated a locus on chromosomal region 3q26-q28. Linkage and disease segregation is explained by a missense mutation c.3614G>A (p.Arg1205His) in eukaryotic translation initiation factor 4-gamma (EIF4G1). Subsequent sequence and genotype analysis identified EIF4G1 c.1505C>T (p.Ala502Val), c.2056G>T (p.Gly686Cys), c.3490A>C (p.Ser1164Arg), c.3589C>T (p.Arg1197Trp) and c.3614G>A (p.Arg1205His) substitutions in affected subjects with familial parkinsonism and idiopathic Lewy body disease but not in control subjects. Despite different countries of origin, persons with EIF4G1 c.1505C>T (p.Ala502Val) or c.3614G>A (p.Arg1205His) mutations appear to share haplotypes consistent with ancestral founders. eIF4G1 p.Ala502Val and p.Arg1205His disrupt eIF4E or eIF3e binding, although the wild-type protein does not, and render mutant cells more vulnerable to reactive oxidative species. EIF4G1 mutations implicate mRNA translation initiation in familial parkinsonism and highlight a convergent pathway for monogenic, toxin and perhaps virally-induced Parkinson disease.

Associations between gut microbiota characteristics and non-motor symptoms following pharmacological and surgical treatments in Parkinson's disease patients.

BACKGROUND: The gut microbiota has been implicated in Parkinson's disease (PD), with alterations observed in microbial composition and reduced microbial species richness, which may influence gastrointestinal symptoms in PD patients. It remains to be determined whether the severity of gastrointestinal symptoms correlates with microbiota variations in PD patients treated pharmacologically or with subthalamic nucleus deep brain stimulation (STN-DBS) therapy. This study aims to explore how these treatments affect gut microbiota and gastrointestinal symptoms in PD, identifying specific microbial differences associated with each treatment modality. METHODS: A total of 42 individuals diagnosed with PD, along with 38 age-matched household control participants, contributed stool samples for microbiota characterization. Differences in the gut microbiota across various groups of PD patients and their households were identified through comprehensive sequencing of the 16S rRNA gene amplicon sequencing. KEY RESULTS: Differences in microbial communities were observed between PD patients and controls, as well as between PD patients receiving pharmacological treatment and those with STN-DBS. Pharmacologically treated advanced PD patients have higher gastrointestinal dysfunctions. Gut microbiota profile linked to STN-DBS and reduced levodopa consumption, characterized by its anti-inflammatory properties, might play a role in diminishing gastrointestinal dysfunction relative to only pharmacological treatments. CONCLUSIONS & INFERENCES: Advanced PD patients on medication exhibit more gastrointestinal issues, despite relatively stable microbial diversity, indicating a complex interaction between gut microbiota, PD progression, and treatment effects. An imbalanced gut-brain axis, particularly due to reduced butyrate production, may lead to constipation by affecting the enteric nervous system, which emphasizes the need to incorporate gut microbiome insights into treatment strategies.

A multi-centre clinico-genetic analysis of the VPS35 gene in Parkinson disease indicates reduced penetrance for disease-associated variants.

BACKGROUND: Two recent studies identified a mutation (p.Asp620Asn) in the vacuolar protein sorting 35 gene as a cause for an autosomal dominant form of Parkinson disease . Although additional missense variants were described, their pathogenic role yet remains inconclusive. METHODS AND RESULTS: We performed the largest multi-center study to ascertain the frequency and pathogenicity of the reported vacuolar protein sorting 35 gene variants in more than 15,000 individuals worldwide. p.Asp620Asn was detected in 5 familial and 2 sporadic PD cases and not in healthy controls, p.Leu774Met in 6 cases and 1 control, p.Gly51Ser in 3 cases and 2 controls. Overall analyses did not reveal any significant increased risk for p.Leu774Met and p.Gly51Ser in our cohort. CONCLUSIONS: Our study apart from identifying the p.Asp620Asn variant in familial cases also identified it in idiopathic Parkinson disease cases, and thus provides genetic evidence for a role of p.Asp620Asn in Parkinson disease in different populations worldwide.

Visual feedback training using WII Fit improves balance in Parkinson's disease.

Postural instability including imbalance is the most disabling long term problem in Parkinson's disease (PD) that does not respond to pharmacotherapy. This study aimed at investigating the effectiveness of a novel visual-feedback training method, using Wii Fit balance board in improving balance in patients with PD. Twenty four patients with moderate PD were included in the study which comprised of a 6-week home-based balance training program using Nintendo Wii Fit and balance board. The PD patients significantly improved their results in Berg Balance Scale, Tinnet's Performance-Oriented Mobility Assessment, Timed Up-and-Go, Sit-to-stand test, 10-Meter Walk test and Activities-specific Balance Confidence scale at the end of the programme. This study suggests that visual feedback training using Wii-Fit with balance board could improve dynamic and functional balance as well as motor disability in PD patients.

Efficacy of deep brain stimulation of the subthalamic nucleus on autonomic dysfunction in patients with Parkinson's disease.

Subthalamic nucleus (STN) deep brain stimulation (DBS) is well established for the treatment of the motor symptoms of Parkinson's disease (PD). However, the effect of STN DBS on autonomic symptoms has not been well studied. We examined 19 patients undergoing STN DBS for PD. The patients were administered a questionnaire to evaluate the pre-operative and post-operative autonomic function. All patients reported a significant post DBS improvement of one or more symptoms of the autonomic dysfunction (urinary and gastrointestinal function). In particular, we have shown the most significant improvement in the urinary function after STN DBS. Further larger studies are required with respect to the effect of STN DBS on the autonomic function.

Ecchordosis physaliphora - a case report and a review of notochord-derived lesions.

Some notochord cells remain along the axis of the vertebral column after embryogenesis. These 'notochordal remnants' have some similarities, but their biological behaviour varies considerably. They can give rise to benign lesions such as ecchordosis physaliphora (EP) and 'benign notochordal cell tumour' (BNCT), or aggressive ones like chordoma. We review the problems of the differential diagnosis of notochordal remnants apropos of a case of the incidental autopsy finding of EP in a 78-year-old man, who died due to heart infarction. The 6-mm asymptomatic gelatinous lesion was fixed to the basilar artery on its ventral aspect. Small EPs can be easily overlooked in autopsy. Ecchordosis physaliphora and intradural chordoma share some similarities that may be misleading and may even result in the wrong diagnosis and therapy. The recently reported new entity BNCT poses a similar problem. We review the literature illustrating the most important features of notochord-derived lesions and discuss the relationships between these lesions with regard to molecular genetics.

Frequency of mutations in PRKN, PINK1, and DJ1 in Patients With Early-Onset Parkinson Disease from neighboring countries in Central Europe.

INTRODUCTION: Approximately 10% of patients with Parkinson disease (PD) present with early-onset disease (EOPD), defined as diagnosis before 50 years of age. Genetic factors are known to contribute to EOPD, with most commonly observed mutations in PRKN, PINK1, and DJ1 genes. The aim of our study was to analyze the frequency of PRKN, PINK1, and DJ1 mutations in an EOPD series from 4 neighboring European countries: Czech Republic, Germany, Poland, and Ukraine. METHODS: Diagnosis of PD was made based on UK Brain Bank diagnostic criteria in departments experienced in movement disorders (1 from Czech Republic, 1 from Germany, 9 from Poland, and 3 from Ukraine). EOPD was defined as onset at or before 50 years of age. Of the 541 patients recruited to the study, 11 were Czech, 38 German, 476 Polish, and 16 Ukrainian. All cohorts were fully screened with Sanger sequencing for PRKN, PINK1, and DJ1 and multiplex ligation-dependent probe amplification for exon dosage. RESULTS: PRKN homozygous or double heterozygous mutations were identified in 17 patients: 1 Czech (9.1%), 1 German (2.6%), 14 Polish (2.9%), and 1 Ukrainian (6.3%). PINK1 homozygous mutations were only identified in 3 Polish patients (0.6%). There were no homozygous or compound heterozygous DJ1 mutations in analyzed subpopulations. One novel variant in PRKN was identified in the Ukrainian series. CONCLUSION: In the analyzed cohorts, mutations in the genes PRKN, PINK1, and DJ1 are not frequently observed.

[Effects of deep brain stimulation on dysfunctions of the autonomic nervous system in Parkinson's disease]. / Wplyw glebokiej stymulacji mózgu na zaburzenia autonomicznego ukladu nerwowego w chorobie Parkinsona.

BACKGROUND: Deep brain stimulation (DBS) for alleviation of motor symptoms in advanced Parkinson's disease (PD) is well established. However, autonomic effects of DBS are less clear. AIM: To review published data on autonomic dysfunctions in DBS-treated patients with PD. METHODS: Medline bibliographic search was performed with the selected relevant keywords, through June 10th, 2010. Thirty three original papers meeting the criteria were identified. RESULTS: Effect of DBS on autonomic dysfunctions observed in PD differs depending on the underlying etiology. DBS of the subthalamic nucleus has no direct effects on cardiovascular functions. Relative improvement results from reduced levodopa intake. Majority of gastrointestinal and urinary tract disorders improve with neurostimulation. However, prolonged body weight gain is observed. Furthermore, urinary tract functions normalize, and in male patients with Parkinson's disease sexual satisfaction increases. Sweating disorders are markedly reduced. On the contrary, chronic stimulation in the area of STN may induce autonomic adverse effects including sialorrhea and urinary retention. In exceptional cases withdrawal of medication before planned DBS surgery may result in life-threatening parkinsonism-hyperpyrexia syndrome. Thus discontinuation of levodopa should be an inpatient procedure. CONCLUSIONS: Selection of patients who undergo DBS should weigh potential benefits and risks resulting from multidirectional effects of neurostimulation on autonomic nervous system.

Assessments of plasma acyl-ghrelin levels in Parkinson's disease patients treated with deep brain stimulation.

Gastrointestinal dysfunction is the most common non-motor symptom in Parkinson's disease (PD) with rates rising as the disease progresses. Deep brain stimulation of subthalamic nucleus (STN DBS) improves motor functions in advanced PD. However, the effect of STN DBS on ghrelin concentration and consequently on motility disturbances as well as body weight is unclear. The objective of this study was to assess acyl-ghrelin levels in comparison to weight in advanced PD patients treated with STN DBS. Plasma concentrations of acyl-ghrelin was measured in 29 PD patients in the fasting state and at 30, 60, 120, and 180 min after a standard meal preoperatively and 3 months after surgery. The level of acyl-ghrelin in PD patients were compared with 30 age and sex-matched healthy controls. We reported that mean plasma acyl-ghrelin levels were decreased in PD patients before STN DBS in fasting (p = 0.0003) and in 30 min postprandial phase (p = 0.04) compared with healthy controls. The plasma acyl-ghrelin levels after STN DBS increased in pre-prandial and postprandial phase in PD patients at the investigated time points. Body weight gained on average 2.33 kg during the first 3 months after surgery. There was no correlation between the acyl-ghrelin plasma levels and BMI. After STN DBS in fasting and postprandial phase plasma acyl-ghrelin levels were increased. The results showed that STN DBS therapy elicited a modification of ghrelin levels, increasing its concentration in pre- and postprandial state. In addition, body weight was increased during 3 months after surgery.

Gene polymorphisms and motor levodopa-induced complications in Parkinson's disease.

OBJECTIVE: The aim of the study was to evaluate the association of individual and combined single-nucleotide polymorphisms in brain-derived neurotrophic factor (BDNF), dopamine transporter (DAT), and catechol-O-methyltransferase (COMT) genes with the occurrence of motor levodopa-induced complications (MLIC) in Parkinson's disease (PD). MATERIALS AND METHODS: We studied 76 patients with PD (MLIC occurred in 56.6%) and 60 controls. Allelic discrimination of rs6265 BDNF (Val66Met), rs397595 DAT (SLC6A3), and rs4680 COMT (Val158Met) genes were genotyped. Odds ratios (OR) and 95% confidence intervals (95% CI) were calculated using multinominal logistic regression. Orthogonal partial least squares (OPLS) analysis and OPLS discriminant analysis (OPLS-DA) were used to analyze qualitative genetic data. RESULTS: The risk of PD in subjects with the AG BDNF genotype was increased sixfold (OR = 6.12, 95% CI = 2.88-13.02, p < .0001), and AG BDNF and AG DAT genotypes were correlated with PD in OPLS-DA (VIP > 1). There were no differences in distributions of BDNF, DAT and COMT genotypes between PD groups with and without MLIC, while OPLS model showed that genotype combination of AG BDNF, AG DAT, and GG COMT was correlated with MLIC and genotypes combination of GG BDNF, AA DAT, and AA COMT with lack of MLIC in PD patients (VIP > 1). CONCLUSIONS: Our results confirmed the association of rs6265 BDNF (Val66Met) with the risk of PD and suggest a synergic effect of rs6265 BDNF (Val66Met), rs397595 DAT (SLC6A3), and rs4680 COMT (Val158Met) polymorphisms on the occurrence of MLIC.

Activities of Daily Living and Their Relationship to Health-Related Quality of Life in Patients with Parkinson Disease After Subthalamic Nucleus Deep Brain Stimulation.

BACKGROUND: Deep brain stimulation (DBS) of the subthalamic nucleus (STN) can reduce motor symptoms in patients with Parkinson disease (PD) and improve their health-related quality of life (HRQoL). The effect of STN DBS on activities of daily living (ADL), an important component of quality of life, is poorly understood. We aimed to investigate effects of STN DBS on HRQoL and ADL in patients with PD. METHODS: HRQoL and ADL were assessed using the following disease-specific and generic questionnaires at baseline and 3, 6, and 12 months after surgery: the Parkinson's Disease Questionnaire 39 (PDQ-39), the Short-Form 36 health survey questionnaire, the World Health Organization Quality of Life Scale-Brief version, the Unified Parkinson's Disease Rating Scale part II, the ADL scale, and the Instrumental Activities of Daily Living scale. RESULTS: We reported significant early improvements (3 months) in the HRQoL and ADL, and these benefits increased over time (6 months); however, further improvement between 6 and 12 months was nonsignificant. Two PDQ-39 subdomains (social support and communications) and a Short-Form 36 health survey questionnaire subdomain (social functioning) showed declines after surgery. Changes in the Instrumental Activities of Daily Living scale were significantly correlated with changes in the PDQ-39 summary index and other PDQ-39 subdomains, including mobility, emotional well-being, social support, and cognition, at all follow-up points. CONCLUSIONS: STN DBS caused a marked improvement in HRQoL at 3 and 6 months; however, HRQoL remained stable at the 12-month postoperative follow-up. Moreover, we have shown a significant correlation between ADL performance and HRQoL after STN DBS.

Genetic variation of Omi/HtrA2 and Parkinson's disease.

Variants in the Omi/HtrA2 gene have been nominated as a cause of Parkinson's disease. This sequencing study of Omi/HtrA2 in 95 probands with apparent autosomal dominant inheritance of Parkinson's disease did not identify any pathogenic mutations. In addition, there was no association between common variations in the Omi/HtrA2 gene and susceptibility to Parkinson's disease in any of our four patient-control series (n=2373). Taken together our results do not support a role for Omi/HtrA2 variants in the pathogenesis of Parkinson's disease.

Dopamine beta-hydroxylase -1021C>T association and Parkinson's disease.

A single nucleotide polymorphism in the promoter region of the dopamine beta-hydroxylase gene (DBH -1021C>T; rs1611115) is reported to regulate plasma enzyme activity levels. This variant has also been the focus of two large association studies in Parkinson's disease yielding conflicting results. We examined this association in four Caucasian patient-control series (n=2696). A modest protective association was observed in the Norwegian series (OR=0.81, p=0.03; n=1676), however, the effect was in the opposite direction in the Polish series (OR=2.01, p=0.01; n=224). No association was observed for DBH -1021C>T with disease susceptibility in the US and Irish series, or combining all four series (OR=0.91, p=0.16, n=2696). We observed a modest association between DBH -1021C>T and AAO in the combined series (p=0.01). Taken together, these findings indicate that DBH -1021C>T does not play a major role in the pathogenesis of Parkinson's disease.

Study of Cu chemical state inside single neurons from Parkinson's disease and control substantia nigra using the micro-XANES technique.

Parkinson's disease (PD) is referred to as idiopathic disorder, which means that its causes have not been found yet. However, a few processes such as oxidative stress, protein aggregation and mitochondrial dysfunction are suspected to lead to the atrophy and death of substantia nigra (SN) neurons in case of this neurodegenerative disorder. Cu is a trace element whose role in the pathogenesis of PD is widely discussed. The investigation of Cu oxidation state inside single nerve cells from SN of PD and control cases may shed some new light on the role of this element in PD. The differences in Cu chemical state were investigated with the use of X-ray absorption near edge structure (XANES) spectroscopy. The least-square fitting method was applied for the analysis of XANES spectra. The comparison of the positions of white line, multiple scattering and pre-edge peak maximum at the energy scale did not reveal the existence of differences in Cu chemical state between PD and control samples. However, it was found that most of the Cu inside SN neurons occurs in tetrahedral environment and probably as Cu(II).

Increased synphilin-1 expression in human elderly brains with substantia nigra Marinesco bodies.

The aim of the present study was to examine the expression of synphilin-1, alpha-synuclein, and tyrosine hydroxylase in human elderly brains and the incidence of Marinesco bodies (MBs, intranuclear inclusions) in the neuromelanin-containing substantia nigra neurons. The brains of twenty-two individuals without clinical signs and symptoms of parkinsonism and dementia and an additional two parkinsonian patients were dissected and subjected to histopathological examination and western blotting. Ubiquitin-positive and agr;-synuclein-negative MBs were found in 0.84-9.45% of the nigral neurons from brains of 15 healthy individuals and both parkinsonian patients. The frequency of pigmented nigral neurons containing MBs was positively correlated with age. The levels of tyrosine hydroxylase in the caudate nucleus and putamen decreased with age, and were inversely correlated with the MB frequency. The level of synphilin-1 in the caudate nucleus was positively correlated both with age and the MBs. Additionally, the MB appearance was correlated with synphilin-1 level in the substantia nigra. No significant correlation between alpha-synuclein expression and age or MBs was found. Our results suggest that synphilin-1 expression increases with aging. Further studies on expression of this protein in elderly brains are warranted.