Polymorphisms in the receptor for GDNF (RET) are not associated with Parkinson's disease in Southern Germany.

The aetiology of the selective neurodegeneration in Parkinson's disease (PD) is still unknown. Neurotrophic factors, e.g. glial cell line-derived neurotrophic factor (GDNF), have been shown to promote survival of dopaminergic neurons. Interestingly, aged mice lacking GDNF-receptor (RET) in their dopaminergic neurons show a phenotype similar to presymptomatic PD. We therefore were interested whether polymorphisms in the RET gene were associated with increased PD risk. Analyzing 25 SNPs in the RET region in 340 Southern German PD patients and 340 age- and sex-matched controls from Southern Germany (KORA S4), we did not find any significant association with PD, suggesting that the equilibrium of trophic factors in PD might be disturbed on other levels than the genomic encoding.

A longitudinal study of tremor frequencies in Parkinson's disease and essential tremor.

OBJECTIVE: There is evidence that the tremor frequency in essential tremor (ET) decreases with time. Longitudinal studies on the evolution of tremor frequencies in Parkinson's disease (PD) have so far not been published. Here, we present a longitudinal analysis of tremor frequencies in PD and ET. METHODS: We analyzed the standardized accelerometric and electromyographic tremor recordings of 53 patients with PD and 38 patients with ET who underwent repeated routine tremor recordings between 1991 and 2002. RESULTS: In an average follow-up period of 44.9 months in PD and 50.6 months in ET, the average number of tremor recordings was 3.3 in PD and 3.7 in ET. In both disorders, tremor frequencies tended to decrease with time. The average annual decrease of the tremor frequency was 0.09 Hz/year in Parkinsonian rest tremor, 0.08 Hz/year in Parkinsonian postural tremor and 0.12 Hz/year in ET. CONCLUSIONS: The tremor frequency decreases with time in both PD and ET. The similarity of this decrease in PD and ET may point to a common underlying pathophysiological mechanism. SIGNIFICANCE: Decreasing tremor frequencies with time may be functionally important by inducing larger tremor amplitudes due to the low-pass filtering properties of muscles and limbs.

Parkin modulates gene expression in control and ceramide-treated PC12 cells.

Mutations in the parkin gene cause autosomal-recessive early-onset parkinsonism as a result of the degeneration of mesencephalic dopaminergic neurons. In cell culture models, parkin expression has been shown to protect against cell death mediated by the sphingolipid ceramide. To determine whether the antiapoptotic effect of parkin involves changes in gene expression, we used Affymetrix oligonucleotide microarrays to analyse gene expression in stably transfected PC12 cells which conditionally overexpress parkin, that were treated or not with C2-ceramide. Overexpression of parkin and ceramide treatment both modulated gene expression. A number of the genes upregulated in the presence of ceramide, and modulated by parkin, were associated with apoptosis or cellular stress reactions. We validated the upregulation of four such genes (CHK, EIF4EBP1, GADD45A and PTPN-5) by real-time PCR after 3, 6, 9 and 12 h of ceramide treatment in cells that overexpressed parkin or not. All were upregulated 2 to 11-fold, 3 and 6 h after application of ceramide. Parkin overexpression reduced the upregulation of EIF4EBP1, GADD45A and PTPN-5, but only at 6 h. These results suggest that, in this assay, the cytoprotective effect of parkin might result not only from its E3-ligase activity, but also from direct or indirect modulation of gene expression in a time-dependent manner.

On studentising and blocklength selection for the bootstrap on time series.

For independent data, non-parametric bootstrap is realised by resampling the data with replacement. This approach fails for dependent data such as time series. If the data generating process is at least stationary and mixing, the blockwise bootstrap by drawing subsamples or blocks of the data saves the concept. For the blockwise bootstrap a blocklength has to be selected. We propose a method for selecting the optimal blocklength. To improve the finite size properties of the blockwise bootstrap, studentised statistics is considered. If the statistic can be represented as a smooth function model this studentisation can be approximated efficiently. The studentised blockwise bootstrap method is applied for testing hypotheses on medical time series.

The effect of carpal tunnel release on median nerve flattening and nerve conduction.

Carpal tunnel syndrome (CTS) is the most common entrapment neuropathy and extensive surveys have been given on the time course of electrophysiological findings pre- and postoperatively. In patients with clinical and electrophysiological confirmed diagnosis of CTS surgical decompression of the carpal tunnel is a first line treatment and has proven to be successfull in 70 to 90% of all cases. The objective of this work was to study the morphological changes of the median nerve after endoscopic release of the carpal tunnel. We used high resolution ultrasound to quantify flattening of the median nerve and to calculate a flattening ratio before endoscopic release as well as 2 weeks and 3 months postoperatively. Ten patients with clinical and electrophysiological confirmed CTS were included in the study. There was significant normalization of the calculated flattening ratio of the median nerve already 2 weeks after surgical release, whereas nerve conduction studies needed a longer period of time to normalize and thus were still abnormal 3 months postoperatively. We conclude that ultrasound is a simple and excellent objective method for visualizing the morphological recovery of the median nerve very early after decompression surgery. In complex cases with unsatisfactory outcome ultrasonography may prove useful in confirming successfull or failed decompression of the median nerve.

Parkin interacts with the proteasome subunit alpha4.

Mutations in the parkin gene encoding an E3 ligase are responsible for autosomal recessive Parkinson's disease. Putative parkin substrates and interacting partners have been identified, but the molecular mechanism underlying parkin-related neurodegeneration is still unclear. We have identified the 20S proteasomal subunit alpha4 (synonyms: PSMA7, XAPC7, subunit alpha type 7) as a new interacting partner of parkin. The C-terminal IBR-RING domain of parkin and the C-terminal part of alpha4 were essential for the interaction. Biochemical studies revealed that alpha4 was not a substrate for parkin-dependent ubiquitylation. Putative functions of the interaction might therefore be substrate presentation to the proteasome or regulation of proteasomal activity. Full-length parkin and parkin lacking the N-terminal ubiquitin-like domain slightly increased the proteasomal activity in HEK 293T cells, in line with the latter hypothesis.

Ultrastructural localization of parkin in the rat brainstem, thalamus and basal ganglia.

The parkin gene encodes a 52 kd putative E3 ubiquitin-protein ligase involved in an autosomal recessive form of early onset parkinsonism. Parkin ultrastructural localization was studied by immunohistochemistry in the adult rat brain and in a parkin inducible PC12 cell line (HS22). In the rat brain, parkin immunoreactivity was detected in neuronal and glial cell bodies and in nerve processes. In the neurons, it was mostly localized on the periphery of large vesicles, some rare mitochondria and endoplasmic reticulum in the cell bodies, and on the periphery of large vesicles in the dendrites and terminals of the neurons. In addition, parkin immunoreactivity was also found around synaptic vesicles in the presynaptic elements of some axons. In HS22 cells over-expressing parkin, the distribution of the protein was similar to that observed in the perikarya of the labeled neurons.

Tremor analysis in two normal cohorts.

OBJECTIVE: Quantitative tremor analyses using almost identical methods were compared between two independent large normal cohorts, to separate robust measures that may readily be used diagnostically from more critical ones needing lab-specific normalization. METHODS: Hand accelerometry and surface EMG from forearm flexors and extensors were recorded with (500 and 1000 g) and without weight loading under postural conditions in 117 and 67 normal volunteers in two different specialty centers for movement disorders in Germany. RESULTS: Tremor amplitude (total power) and frequency fell within a similar range but differed significantly. A significant reduction of tremor frequency under 1000 g weight load (>1 Hz), and a lack of rhythmic EMG activity at the tremor frequency in around 85-90% of the recordings were robust findings in both centers. CONCLUSIONS: The differences in frequency and total power indicate that these measures critically depend on the details of the recording conditions being slightly different between the two centers. Thus each lab needs to establish its own normative data. We estimate that at least 25 normal subjects have to be recorded to obtain normal values. The reduction of tremor frequency under load and lacking tremor-related EMG activity were well reproducible allowing a differentiation of physiological from low amplitude pathological tremor. SIGNIFICANCE: This study provides a framework for more standardized tremor analyses in clinical neurophysiology.

Dynamic synchronisation of central oscillators in essential tremor.

OBJECTIVE: Coherence analysis of electromyography (EMG) signals in essential tremor (ET) suggests that tremor in the right and left arm is induced by independent central oscillators. The sensorimotor cortex seems to be part of the tremor-generating neuronal network in ET. Here, we investigated using electroencephalography (EEG) whether the independence of central oscillators in ET is supported by the analysis of cortical activity. METHODS: In 8 patients with ET, bilateral hand tremor was activated by wrist extension. EMGs from the wrist flexors and extensors were recorded simultaneously with an EEG. EEG-EMG coherence was estimated for 74 epochs of 60 s duration. RESULTS: In 42.6% of the cases, EEG-EMG coherence at the tremor frequency existed only with the contralateral sensorimotor cortex. However, 21.6% of the tremor-EMGs were coherent with EEG activity over both the contralateral and ipsilateral sensorimotor cortex. Bilateral and exclusively contralateral EEG-EMG coherence could alternate within the same recording. Bilateral EEG-EMG coherence was associated with increased right-left EEG-EEG coherence, increased right-left EMG-EMG coherence as well as with increased tremor strength. CONCLUSIONS: In ET, central oscillators in the right and left brain are not entirely independent of each other. They may dynamically synchronise, presumably by interhemispheric coupling via the corpus callosum.

Effect of chronic bilateral subthalamic nucleus (STN) stimulation on postural control in Parkinson's disease.

Postural instability is one of the most incapacitating factors in Parkinson's disease (PD). The underlying deficits and the effects of treatment are still not well understood. The aims of the present study were: (i) to identify abnormalities of postural control in PD patients during unperturbed stance and externally perturbed stance (anterior-posterior tilts of the support surface and of the visual scene); (ii) to assess the effects of L-dopa medication and subthalamic nucleus (STN) stimulation on posture control; and (iii) to characterize potential differential or additive effects of both treatments. Eight PD patients under chronic STN stimulation were investigated and compared with 10 normal controls. The assessment was performed in a crossover design (+/- STN stimulation, +/- L-dopa). During unperturbed stance, we recorded measures of spontaneous sway in terms of displacement, velocity and frequency of the centre of pressure (COP), lower body (LB) and upper body (UB) excursions. In addition, inter-segmental UB-LB coupling was investigated as a measure of axial stiffness. All these measures were abnormally large in patients OFF treatment. Under L-dopa treatment, the velocity, frequency and coupling measures were reduced, whereas sway amplitude increased. Very similar effects were obtained under STN stimulation, and these effects became more pronounced in the combined treatment condition. In these data, reduction of inter-segmental coupling correlated with increase in sway amplitude. The finding suggests that axial stiffness reduction under treatment revealed a treatment- resistant deficit in the sensorimotor postural control loop. However, these two effects did not correlate with the motor subscores of the unified Parkinson's disease rating scale (UPDRS), which indicates that they are of minor functional relevance for posture control. A frequency peak in the COP excursions at 0.7-1.1 Hz, which we take to indicate a resonance behaviour of the postural control loop, became reduced under therapy. The reduction of this peak did correlate with most improvements in the UPDRS under therapy. Support surface tilt revealed that an UB righting on the LB segment, which is present in normal controls, is missing in the patients. The postural responses to visual tilt were abnormally large in patients, independent of whether the support was stable or slightly moving, while the control subjects clearly profited from a stable support. This finding suggests that PD patients lack the ability of normal subjects to use sensory or cognitive information when suppressing the destabilizing effect of visual tilt. These abnormal tilt reactions of the patients were resistant to treatment with L-dopa, STN stimulation and a combination of the two. Overall, the effects of STN stimulation on posture control essentially paralleled those of L-dopa during both unperturbed and externally perturbed stance.

New parkin mutations and atypical phenotypes in families with autosomal recessive parkinsonism.

The frequency of parkin mutations was evaluated in 30 families of highly diverse geographic origin with early-onset autosomal recessive parkinsonism. Twelve different mutations, six of which were new, were found in 10 families from Europe and Brazil. Patients with parkin mutations had significantly longer disease duration than patients without the mutation but with similar severity of disease, suggesting a slower disease course. Two patients with parkin mutations had atypical clinical presentation at onset, with predominant tremor when standing.

Estimation of delay times in biological systems.

The problem of delay time estimation in biological systems is addressed with the focus on practical applicability of methods. Four delay time estimators are described: a cross correlation method and three increasingly sophisticated interpretations of the phase spectrum, ranging from a pointwise interpretation of the phase spectrum in terms of a delay to a Hilbert transform method. The four methods are compared through simulation studies showing that, in general, the Hilbert transform method performs best. The methods are then used to estimate delay times in three physiological systems: vestibular stimulation, cerebral autoregulation, and human orthostatic tremor. In all three cases, the Hilbert transform method yields the best results, leading in some cases to physiologically more sensible interpretations of experiments than the other methods.

Essential tremor and cerebellar dysfunction: abnormal ballistic movements.

BACKGROUND: Clinical characteristics reminiscent of cerebellar tremor occur in patients with advanced essential tremor. Ballistic movements are known to be abnormal in cerebellar disease. The hypothesis was proposed that ballistic movements are abnormal in essential tremor, reflecting cerebellar dysfunction. OBJECTIVE: To elucidate the role of the cerebellum in the pathophysiology of essential tremor. METHODS: Kinematic parameters and the triphasic electromyographic (EMG) components of ballistic flexion elbow movements were analysed in patients assigned to the following groups: healthy controls (n = 14), pure essential postural tremor (ET(PT); n = 17), and essential tremor with an additional intention tremor component (ET(IT); n = 15). RESULTS: The main findings were a delayed second agonist burst (AG(2)) and a relatively shortened deceleration phase compared with acceleration in both the essential tremor groups. These abnormalities were most pronounced in the ET(IT) group, which had additional prolongation of the first agonist burst (AG(1)) and a delayed antagonist burst (ANT). CONCLUSIONS: Abnormalities of the triphasic pattern and kinematic parameters are consistent with a disturbed cerebellar timing function in essential tremor. These abnormalities were most pronounced in the ET(IT) group. The cerebellar dysfunction in essential tremor could indicate a basic pathophysiological mechanism underlying this disorder. ET(PT) and ET(IT) may represent two expressions within a continuous spectrum of cerebellar dysfunction in relation to the timing of muscle activation during voluntary movements.

Hyperglycemia in patients with focal cerebral ischemia after intravenous thrombolysis: influence on clinical outcome and infarct size.

The aim of the present prospective study was to investigate whether hyperglycemia influences the clinical outcome or the infarct size after intravenous thrombolysis of focal cerebral ischemia. A consecutive series of hyperglycemic (n = 14) and normoglycemic patients (n = 17) with acute focal cerebral ischemia (<3 h) in the middle cerebral artery (MCA) territory received rtPA (0.9 mg/kg body weight) intravenously. Clinical outcome was measured using the NIH Stroke Score on admission and was followed up until day 28. Infarct volume was measured by diffusion-weighted MR imaging on admission, on days 3 and 7. There was a significantly better neurological outcome on day 28 in the normoglycemic patients than in the hyperglycemic group (NIH SS 4.0 versus 7.4; p < 0.05). The infarction volume increased significantly in the hyperglycemic patients Delta = 39.9 plus minus 17.4% compared to normoglycemic patients Delta = 27.1 plus minus 14.1% (p < 0.05). The present study suggests that hyperglycemia in patients with a focal MCA ischemia can cause a worse clinical outcome despite recanalization of the occluded vessel by thrombolysis therapy. This correlates with a markedly larger increase of the infarction volume in the hyperglycemic group. These results may be explained by an accentuated lactate accumulation and pH decrease by elevated energy levels which cannot be compensated by restoration of blood flow alone.

Up-regulation of D3 dopaminergic receptor mRNA in the core of the nucleus accumbens accompanies the development of seizures in a genetic model of absence-epilepsy in the rat.

The basal ganglia system is thought to play a key role in the control of absence-seizures and there is ample evidence that epileptic seizures modify brain dopamine function. We recently reported that local injections of dopamine D1 or D2 agonists in the core of the nucleus accumbens suppressed absence-seizures in a spontaneous, genetic rodent model of absence-epilepsy whereas injections of D1 or D2 antagonists had aggravating effects. These findings raised the possibility that the dopaminergic system may be altered in absence-epilepsy prone rats. Therefore, we studied by in situ hybridization histochemistry the expression of pre- and postsynaptic components of the dopaminergic system in this strain of rats. When compared to non-epileptic control rats, epileptic rats displayed no change in the expression of mRNAs coding for the neuronal dopaminergic markers (tyrosine hydroxylase, membraneous and vesicular dopamine transporters). In addition, there was no difference between the two strains concerning the expression of the dopamine receptor transcripts D1, D2 and D5. In adult absence-epilepsy prone rat with an overt epileptic phenotype, however, an elevated level of D3 mRNA expression was observed in neurons of the core of the nucleus accumbens (+23% increase in silver grain density compared to non-epileptic control rats). D3 transcripts were not increased in juvenile epileptic rats without seizures. These findings suggests that up-regulation of D3 receptor mRNA is part of the epileptic phenotype in absence-epilepsy prone rats. Its localization in the core of the nucleus accumbens bears close resemblance to the dopamine-sensitive antiepileptic sites in ventral striatum and further support the involvement of ventral structures of the basal ganglia system in the control of absence-seizures.

Valproic acid triggers acute rhabdomyolysis in a patient with carnitine palmitoyltransferase type II deficiency.

A 47-year-old man suffering from a bipolar disorder and intermittent myoglobinuria presented with acute rhabdomyolysis with renal failure after starting therapy with valproic acid. On morphological examination, skeletal muscle revealed increased lipid storage. Biochemically, decreased enzyme activity of carnitine palmitoyltransferase (CPT) type II with carnitine levels in the lower limit was found. Genetic analysis detected the common Ser113Leu substitution on one allele of the CPT2 gene. We conclude that valproic acid should be avoided in patients with CPT type II deficiency.

Pseudo-dominant inheritance and exon 2 triplication in a family with parkin gene mutations.

The authors report an Italian family with pseudo-dominant inheritance of parkinsonism attributable to parkin gene mutations. The father (disease onset at age 57 years) was homozygous for a triplication of exon 2 that is so far unique. The unaffected mother was heterozygous for deletions of exons 3 and 4, and the son (onset at age 31 years) was a compound heterozygote carrying both mutations. Thus, pseudo-dominant inheritance of parkin gene mutations has to be considered in early-onset parkinsonism, and sensitive screening techniques, such as semiquantitative multiplex PCR, should be applied.

Three parkin gene mutations in a sibship with autosomal recessive early onset parkinsonism.

The objective was to describe a family with autosomal recessive, early onset parkinsonism, with affected siblings carrying three different exon rearrangements in the parkin gene. The living affected siblings were personally examined. Molecular genetic analyses included exon dosage of the parkin gene using a semiquantitative multiplex polymerase chain reaction (PCR) protocol and haplotype analysis. The index case was a compound heterozygote with a deletion of exon 5 and a duplication of exon 3. His affected sister was a compound heterozygote for a deletion of exon 5 and a deletion of exons 3-9. Haplotype analysis confirmed the presence of three mutant alleles at the parkin locus. The phenotype was early onset parkinsonism with marked response to levodopa, and a very slow, prolonged course. In conclusion, the frequency of mutations in the parkin gene in certain populations might be high enough to cause allelic heterogeneity in the same sibship.