Reinforcement learning of self-regulated sensorimotor ß-oscillations improves motor performance.

Self-regulation of sensorimotor oscillations is currently researched in neurorehabilitation, e.g. for priming subsequent physiotherapy in stroke patients, and may be modulated by neurofeedback or transcranial brain stimulation. It has still to be demonstrated, however, whether and under which training conditions such brain self-regulation could also result in motor gains. Thirty-two right-handed, healthy subjects participated in a three-day intervention during which they performed 462 trials of kinesthetic motor-imagery while a brain-robot interface (BRI) turned event-related ß-band desynchronization of the left sensorimotor cortex into the opening of the right hand by a robotic orthosis. Different training conditions were compared in a parallel-group design: (i) adaptive classifier thresholding and contingent feedback, (ii) adaptive classifier thresholding and non-contingent feedback, (iii) non-adaptive classifier thresholding and contingent feedback, and (iv) non-adaptive classifier thresholding and non-contingent feedback. We studied the task-related cortical physiology with electroencephalography and the behavioral performance in a subsequent isometric motor task. Contingent neurofeedback and adaptive classifier thresholding were critical for learning brain self-regulation which, in turn, led to behavioral gains after the intervention. The acquired skill for sustained sensorimotor ß-desynchronization correlated significantly with subsequent motor improvement. Operant learning of brain self-regulation with a BRI may offer a therapeutic perspective for severely affected stroke patients lacking residual hand function.

Neurostimulation for Parkinson's disease with early motor complications.

BACKGROUND: Subthalamic stimulation reduces motor disability and improves quality of life in patients with advanced Parkinson's disease who have severe levodopa-induced motor complications. We hypothesized that neurostimulation would be beneficial at an earlier stage of Parkinson's disease. METHODS: In this 2-year trial, we randomly assigned 251 patients with Parkinson's disease and early motor complications (mean age, 52 years; mean duration of disease, 7.5 years) to undergo neurostimulation plus medical therapy or medical therapy alone. The primary end point was quality of life, as assessed with the use of the Parkinson's Disease Questionnaire (PDQ-39) summary index (with scores ranging from 0 to 100 and higher scores indicating worse function). Major secondary outcomes included parkinsonian motor disability, activities of daily living, levodopa-induced motor complications (as assessed with the use of the Unified Parkinson's Disease Rating Scale, parts III, II, and IV, respectively), and time with good mobility and no dyskinesia. RESULTS: For the primary outcome of quality of life, the mean score for the neurostimulation group improved by 7.8 points, and that for the medical-therapy group worsened by 0.2 points (between-group difference in mean change from baseline to 2 years, 8.0 points; P=0.002). Neurostimulation was superior to medical therapy with respect to motor disability (P<0.001), activities of daily living (P<0.001), levodopa-induced motor complications (P<0.001), and time with good mobility and no dyskinesia (P=0.01). Serious adverse events occurred in 54.8% of the patients in the neurostimulation group and in 44.1% of those in the medical-therapy group. Serious adverse events related to surgical implantation or the neurostimulation device occurred in 17.7% of patients. An expert panel confirmed that medical therapy was consistent with practice guidelines for 96.8% of the patients in the neurostimulation group and for 94.5% of those in the medical-therapy group. CONCLUSIONS: Subthalamic stimulation was superior to medical therapy in patients with Parkinson's disease and early motor complications. (Funded by the German Ministry of Research and others; EARLYSTIM ClinicalTrials.gov number, NCT00354133.).

Cellular uptake of cationic gadolinium-DOTA peptide conjugates with and without N-terminal myristoylation.

Cellular and nuclear uptake of dual labelled conjugates could be of great value for chemotherapy and cancer diagnostics. Therefore we designed conjugates in which gadolinium (Gd)-1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid (DOTA), a contrast agent for magnetic resonance imaging and fluorescein isothiocyanate (FITC), a fluorescence marker were coupled to membrane translocation sequences (MTS). The MTSs we employed were the third helix of the Antennapedia homeodomain, the HIV-1 Tat peptide and the N-myristoylated HIV-1 Tat peptide. We used confocal laser scanning microscopy, fluorescence activated cell sorting, magnetic resonance imaging (MRI) and viability tests to examine the cellular and nuclear uptake of these conjugates into U373 glioma cells, as well as their cytotoxic effects. We found that the Antennapedia conjugate was taken up by no more than 20% of the cells. The HIV-1 Tat conjugate showed even lower uptake into less than 3% of cells. Interestingly, N-myristoylation of the HIV-1 Tat conjugate drastically improved its cellular uptake. Up to 70% of cells showed cellular and nuclear uptake of the N-myristoylated HIV-1 Tat conjugate. Conjugate cytotoxicity appears to correlate with cellular uptake.

Novel cell nucleus directed fluorescent tetraazacyclododecane-tetra-acetic acid compounds.

Peptide conjugates derived from the SV 40 T antigen nuclear localisation sequence (NLS) have been successfully used to translocate both fluorescein isothiocyanate (FITC) and Gadolinium (Gd)-1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid (DOTA) into the cytoplasm and nucleus of glioma cells. However, uptake occurred only in up to 35% of cells. To improve cellular uptake, we designed three novel FITC-labelled Gd-DOTA conjugates. In the first conjugate, the commonly used Gd-DOTA-complex was coupled to the nuclear localization sequence (NLS) of the Simian Virus (SV) 40 T antigen alone as a control. In the second conjugate, the Gd-DOTA-coupled SV 40 T antigen NLS was elongated by the HIV-1 tat peptide (HIV-NLS). A third conjugate, in which the Gd-DOTA-complex was coupled to the SV 40 T antigen NLS elongated by a peptide containing seven arginines and six aminohexanoic acids (Ahx6R7) was also synthesized (AHX-NLS). By means of confocal laser scanning microscopy, fluorescence activated cell sorting, magnetic resonance imaging (MRI) and viability tests we were able to demonstrate that the first conjugate containing only the NLS of the SV 40 T antigen stained the nuclei of no more than 10-12% of U373 and LN18 glioma cells, resulting in low signal intensity in MRI. The stained cells remained viable. After incubation with conjugates HIV-NLS and AHX-NLS the nuclei of up to 73% of U373 and LN18 glioma cells were stained. This was associated with high signal intensity in MRI and cell death. As previously shown, the gadolinium ion reduces cellular uptake of DOTA conjugates. To confirm this, the conjugates were produced with or without gadolinium. The gadolinium-free DOTA conjugates showed a higher cellular uptake rate and an increased cytotoxic potential.

Novel dual labelled nucleus-directed conjugates containing correct and mutant nuclear localisation sequences.

Gadolinium (Gd)-1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid (DOTA) is commonly used as contrast agent in magnetic resonance imaging (MRI), but cannot enter the cytoplasm or cell nucleus. We designed a tetrapeptide carrying fluorescein isothiocyanate (FITC) and Gd-DOTA. This conjugate was coupled to the nuclear localisation sequence (NLS) of the Simian Virus (SV) 40 T antigen elongated by four arginines. In a second conjugate one lysine of the original SV 40 T antigen NLS was replaced by threonine. An FITC-labelled DOTA-tetrapeptide conjugate lacking the NLS peptide served as a negative control. We tried to achieve sequence specific entry of the Gd-DOTA-complex into the cytoplasm and nucleus of human U373 and LN18 glioma cells. Using confocal laser scanning microscopy (CLSM), fluorescence activated cell sorting (FACS), magnetic resonance imaging (MRI) and viability tests we found that both NLS conjugates stained the cell nuclei of U373 and LN18 glioma cells, represented also by a rise in signal intensity compared to the native control in MRI. The majority of stained cells remained viable. All conjugates were also produced without Gd. The Gd-free DOTA-conjugates showed an increase in cellular uptake rate. Conjugate cytotoxicity correlated closely to cellular uptake. Gd-containing DOTA-conjugates directed to the cytoplasm or the nucleus may be the basis for the development of novel diagnostic agents.

A novel polyarginine containing Smac peptide conjugate that mediates cell death in tumor and healthy cells.

The seven N-terminal amino acids AVPIAQK (SmacN7) of the mitochondrial protein Smac (second mitochondria-derived activator of caspase) promote caspase activation by binding specifically to inhibitor of apoptosis proteins (IAPs) and blocking their inhibitory activity. SmacN7 cannot pass through the cell membrane, but to be of therapeutic use it would be essential for it to enter the cell. To achieve transmembrane transport of SmacN7 we coupled it to a novel fluorescein isothiocyanate (FITC)-labelled transmembrane transport peptide RRRRK(FITC)RRRR via ss-alanine to produce the conjugate AVPIAQKssA RRRRK(FITC)RRRR. Because IAPs are much more strongly expressed in the cytoplasm of tumor cells, we expected this conjugate to produce staining of the cytoplasm, and for this to be stronger in tumor cells than in healthy cells. Surprisingly, we found strong nuclear uptake of the Smac conjugate and of the transport peptide alone without subsequent release in both tumor cells and healthy cells from the bladder, prostate, and brain. This was accompanied by cell death. In contrast to expectations, it appears that the apoptotic effects observed do not result from the SmacN7 cargo alone.

Correlation between neonatal calf diarrhea and the level of maternally derived antibodies.

Passively acquired antibodies through colostrum will protect calves against etiological agents of neonatal calf diarrhea. Among them enteric diseases due to strains of Enterotoxigenic Escherichia coli (ETEC) are the most commonly occurring form of colibacillosis in newborn calves. Specific antibodies against whole ETEC cells and total immunoglobulin G in dam serum, colostrum and calf serum were determined. There were significant differences (P=0.0005) between antibody titers in normal and diarrheic groups, in which diarrheic group had a higher titer. Total IgG concentration in diarrheic calves (20.86 ± 0.49), their dams (23.48 ± 0.54) and colostrum (33.40 ± 0.50) was less than normal group (P=0.0005). There was a highly significant positive correlation between dam total IgG with calf total IgG (r=0.022; ratio=52.11). Colostral anti-E. coli antibody had a highly significant positive correlation with anti-E. coli in calf serum (r=0.345; ratio=0.62). Anti-E. coli antibody in calf serum had a highly significant negative correlati with total IgG of dam serum, colostrum and calf serum. While the level of anti-E. coli antibodies in diarrheic group was considerably higher than normal group, our findings reported here are in agreement that immunity to diarrhea also might be correlated with maternal cells or cellular components as well as cytokines which are transferred by colostrum to neonatal calves. Nevertheless, the level of maternally derived antibodies is a promising indicator for passive immunity and protection against diarrhea in neonatal calves.

Functional regeneration of the axotomized auditory nerve with combined neurotrophic and anti-inhibitory strategies.

Injury to the mammalian auditory nerve is associated with a lack of long-distance elongation and leads to definitive loss of the hearing function. To overcome this central nervous system typical lack of functional regeneration, a combined neurotrophic and antiinhibitory treatment is applied. After complete unilateral sectioning of the auditory nerve in adult rats a combination of the Nogo-A inhibitor IN-1 and the neurotrophic factor NT-3 is administrated intrathecally into the cerebellopontine angle for one week. Functional regeneration is evaluated by measuring auditory brainstem evoked potentials for a follow-up period of up to three months. After treatment, up to forty percent of the animals showed a second vertex-positive wave in the auditory brainstem evoked potentials which occurred between three to four weeks after sectioning and remained stable during the follow-up period. A limited degree of functional regeneration of the axotomized auditory nerve is possible after application of IN-1 and NT-3. For additional improvement of functional results further investigations on combined treatments with scar reducing agents, neurotrophic factors and neuroprotective drugs remain necessary.

Electrically evoked hearing perception by functional neurostimulation of the central auditory system.

Perceptional benefits and potential risks of electrical stimulation of the central auditory system are constantly changing due to ongoing developments and technical modifications. Therefore, we would like to introduce current treatment protocols and strategies that might have an impact on functional results of auditory brainstem implants (ABI) in profoundly deaf patients. Patients with bilateral tumours as a result of neurofibromatosis type 2 with complete dysfunction of the eighth cranial nerves are the most frequent candidates for auditory brainstem implants. Worldwide, about 300 patients have already received an ABI through a translabyrinthine or suboccipital approach supported by multimodality electrophysiological monitoring. Patient selection is based on disease course, clinical signs, audiological, radiological and psycho-social criteria. The ABI provides the patients with access to auditory information such as environmental sound awareness together with distinct hearing cues in speech. In addition, this device markedly improves speech reception in combination with lip-reading. Nonetheless, there is only limited open-set speech understanding. Results of hearing function are correlated with electrode design, number of activated electrodes, speech processing strategies, duration of pre-existing deafness and extent of brainstem deformation. Functional neurostimulation of the central auditory system by a brainstem implant is a safe and beneficial procedure, which may considerably improve the quality of life in patients suffering from deafness due to bilateral retrocochlear lesions. The auditory outcome may be improved by a new generation of microelectrodes capable of penetrating the surface of the brainstem to access more directly the auditory neurons.

Trigeminocardiac reflex during skull base surgery: mechanism and management.

BACKGROUND: We study the occurrence and management of the trigeminocardiac reflex (TCR) during neurosurgical procedures for lesions of the skull base. METHOD: Two hundred patients underwent neurosurgical procedures for various skull base lesions and were evaluated retrospectively for the occurrence of the TCR during surgery. This phenomenon was defined as the onset of bradycardia lower than 60 beats/minute and hypotension with a drop in mean arterial blood pressure of 20% or more due to intra-operative manipulation or traction on the trigeminal nerve. FINDINGS: Sixteen patients (8%) had a TCR intra-operatively (7 vestibular schwannomas, 5 sphenoid wing meningiomas, 3 petroclival meningiomas, 1 intracavernous epidermoid cyst). In all 16 patients with a TCR the postoperative courses presented no complications that could be directly related to this intra-operative phenomenon. CONCLUSIONS: Due to the intracranial course of the trigeminal nerve several surgical procedures at the anterior, middle and posterior skull base may elicit the trigeminocardiac reflex. Continuous monitoring of hemodynamic parameters allows the surgeon to interrupt surgical manoeuvres immediately upon the occurrence of the TCR. This technique is sufficient for the heart rate and the arterial blood pressure to return to normal levels without the necessity of additional anticholinergic medication.

Monitoring of blood parameters following anterior cervical fusion.

OBJECT: Both C-reactive protein (CRP) level and erythrocyte sedimentation rate (ESR) were measured prospectively in 51 cases in which uncomplicated cervical anterior fusion was performed. The object of the authors was to quantify the differences in the responses of these parameters recorded in the immediate postoperative period and to determine factors influencing their course. METHODS: Nineteen one-level, 23 two-level, and nine three-level procedures for disc herniation and degenerative disease of the cervical spine were performed in 22 female and 29 male patients (mean age 49.2 years). Blood samples were obtained 1 day before as well as on 10 consecutive days and 3 months following anterior cervical fusion. Serum CRP level was measured using a fluorescence polarization immunoassay and ESR was determined from the same samples. Operative time, the number of blood transfusions, and drugs administered in the postoperative period were recorded. In addition, hemoglobin, hematocrit, red blood cell count, platelet count, white cell count, and axillary body temperature were checked daily. CONCLUSIONS: Monitoring of CRP level is superior to that of ESR for early detection of infections after cervical spine surgery. Although CRP was not related to any of the factors that have been proposed to explain its peak value variance in previous studies, individual acute-phase protein metabolism response to tissue affection appears to be a more decisive element in this respect.

[Three-dimensional-segmentation of the spine with specially adapted tools]. / 3-D-Segmentierung der Wirbelsäule mit speziell angepassten Werkzeugen.

Segmented 3-D data of the spine form the basis for various modern clinical applications. Among these, multisegmental image fusion, image registration and finite element modeling for biomechanical analysis are promising innovative tools capable of facilitating treatment decisions and optimization of individual therapy in the future. However, the complex anatomy of the spine and the often extensive degenerative deformation presenting in clinical practice, generally limit the application of fully automated segmentation. A newly developed software system is presented that meets the complex requirements for image segmentation of the spine through the use of specially adapted interactive tools that take account of its axial skeletal structure. Furthermore, a standardized protocol is introduced that combines the newly developed interactive tools (rotation transformation, warped dissection plane) and standard segmentation tools to permit rapid and accurate segmentation. To date, the software environment presented herein has been applied with success to the segmentation of cervical, thoracic and lumbar spine.

Decoding of motor intentions from epidural ECoG recordings in severely paralyzed chronic stroke patients.

OBJECTIVE: Recently, there have been several approaches to utilize a brain-computer interface (BCI) for rehabilitation with stroke patients or as an assistive device for the paralyzed. In this study we investigated whether up to seven different hand movement intentions can be decoded from epidural electrocorticography (ECoG) in chronic stroke patients. APPROACH: In a screening session we recorded epidural ECoG data over the ipsilesional motor cortex from four chronic stroke patients who had no residual hand movement. Data was analyzed offline using a support vector machine (SVM) to decode different movement intentions. MAIN RESULTS: We showed that up to seven hand movement intentions can be decoded with an average accuracy of 61% (chance level 15.6%). When reducing the number of classes, average accuracies up to 88% can be achieved for decoding three different movement intentions. SIGNIFICANCE: The findings suggest that ipsilesional epidural ECoG can be used as a viable control signal for BCI-driven neuroprosthesis. Although patients showed no sign of residual hand movement, brain activity at the ipsilesional motor cortex still shows enough intention-related activity to decode different movement intentions with sufficient accuracy.

Increased coherence among striatal regions in the theta range during attentive wakefulness.

The striatum, the largest component of the basal ganglia, is usually subdivided into associative, motor and limbic components. However, the electrophysiological interactions between these three subsystems during behavior remain largely unknown. We hypothesized that the striatum might be particularly active during exploratory behavior, which is presumably associated with increased attention. We investigated the modulation of local field potentials (LFPs) in the striatum during attentive wakefulness in freely moving rats. To this end, we implanted microelectrodes into different parts of the striatum of Wistar rats, as well as into the motor, associative and limbic cortices. We then used electromyograms to identify motor activity and analyzed the instantaneous frequency, power spectra and partial directed coherence during exploratory behavior. We observed fine modulation in the theta frequency range of striatal LFPs in 92.5 ± 2.5% of all epochs of exploratory behavior. Concomitantly, the theta power spectrum increased in all striatal channels (P < 0.001), and coherence analysis revealed strong connectivity (coefficients >0.7) between the primary motor cortex and the rostral part of the caudatoputamen nucleus, as well as among all striatal channels (P < 0.001). Conclusively, we observed a pattern of strong theta band activation in the entire striatum during attentive wakefulness, as well as a strong coherence between the motor cortex and the entire striatum. We suggest that this activation reflects the integration of motor, cognitive and limbic systems during attentive wakefulness.

Histopathological features of the brain, liver, kidney and spleen following an innovative polytrauma model of the mouse.

OBJECT: Among the various introduced experimental traumatic brain injury models, there is a clear paucity of proper experimental polytrauma models. To overcome this experimental gap we introduced such a polytrauma model in the mouse including traumatic brain injury. Here, we report on the histopathological features of the brain, lung, kidney, spleen and liver. MATERIALS AND METHODS: 20 male C57BL mice with a mean weight of 23 g were anesthetized with ketamine and xylazine. The anaesthetized animals were subjected to a controlled cortical impact (CCI) over the left parieto-temporal cortex using rounded-tip impounder for application of a standardized brain injury. Following fracture of the right femur using a guillotine, a volume-controlled hemorrhagic shock was induced. The control groups included animals with CCI only (n=20) and animals with femur fracture plus hemorrhagic shock without CCI (n=20). Subjects were sacrified at 96 h following trauma. Brain, lung, kidney, spleen and liver of the animals underwent histopathological examinations. RESULTS: The mortality rate at 96 h was 25% in the polytrauma group versus 10% in the control groups. Within the histopathological investigations, polytraumatized animals differ from those with a single trauma (traumatic brain injury or femur fracture with hemorrhagic shock) with various severity. CONCLUSION: The findings of this study show that such a polytrauma model can be standardized resulting in a reproducible damage. This model fulfills the requirements of a standardized animal model. It allows adequate analogies and inferences to the clinical situation of a polytrauma in humans.

Hematologic and Clinical Aspects of Experimental Ovine Anaplasmosis Caused by Anaplasma ovis in Iran.

BACKGROUND: Anaplasma ovis infections can cause clinical symptoms in acute phase and lead to huge economic losses in flocks. The aim of the present study was to investigate the hematological and parasitological changes in experimental anaplasmosis in sheep with Iranian strain of A. ovis. METHOD: Five male sheep without any blood parasite infection were selected. One hundred ml heparinized blood was collected from splenectomised sheep that showed 6% A. ovis parasitemia. Inoculums of 20 ml blood were administered intravenously to each test animal. Hematological, parasitological and clinical changes of experimental anaplasmosis were studied in 0-38 days post infection. RESULT: Parasitemia was detected 3 days post infection and reached its maximum level on the day 12 of experiment in test animals. Then the parasitemia was declined, but the organism could be found persistently until the last day of study. The red cell counts, packed cell volume and hemoglobin concentration were decreased and mean corpuscular volume was increased significantly during the infection period. Reticulocytosis and basophilic stippling were also detected. No significant changes were observed in total and differential leukocyte count and animal body temperature. CONCLUSION: Experimental A. ovis infection in sheep resulted in marked normocytic normochromic anemia at the beginning of the infection which became macrocytic normochromic by the development of the disease. There were negative correlations between parasitemia and RBC, PCV and Hb values, therefore hematological assessment can be considered as a practical diagnostic tool in ovine anaplasmosis.

Transition from the locked in to the completely locked-in state: a physiological analysis.

OBJECTIVE: To clarify the physiological and behavioral boundaries between locked-in (LIS) and the completely locked-in state (CLIS) (no voluntary eye movements, no communication possible) through electrophysiological data and to secure brain-computer-interface (BCI) communication. METHODS: Electromyography from facial muscles, external anal sphincter (EAS), electrooculography and electrocorticographic data during different psychophysiological tests were acquired to define electrophysiological differences in an amyotrophic lateral sclerosis (ALS) patient with an intracranially implanted grid of 112 electrodes for nine months while the patient passed from the LIS to the CLIS. RESULTS: At the very end of the LIS there was no facial muscle activity, nor external anal sphincter but eye control. Eye movements were slow and lasted for short periods only. During CLIS event related brain potentials (ERP) to passive limb movements and auditory stimuli were recorded, vibrotactile stimulation of different body parts resulted in no ERP response. CONCLUSIONS: The results presented contradict the commonly accepted assumption that the EAS is the last remaining muscle under voluntary control and demonstrate complete loss of eye movements in CLIS. The eye muscle was shown to be the last muscle group under voluntary control. The findings suggest ALS as a multisystem disorder, even affecting afferent sensory pathways. SIGNIFICANCE: Auditory and proprioceptive brain-computer-interface (BCI) systems are the only remaining communication channels in CLIS.

Closing the sensorimotor loop: haptic feedback facilitates decoding of motor imagery.

The combination of brain-computer interfaces (BCIs) with robot-assisted physical therapy constitutes a promising approach to neurorehabilitation of patients with severe hemiparetic syndromes caused by cerebrovascular brain damage (e.g. stroke) and other neurological conditions. In such a scenario, a key aspect is how to reestablish the disrupted sensorimotor feedback loop. However, to date it is an open question how artificially closing the sensorimotor feedback loop influences the decoding performance of a BCI. In this paper, we answer this issue by studying six healthy subjects and two stroke patients. We present empirical evidence that haptic feedback, provided by a seven degrees of freedom robotic arm, facilitates online decoding of arm movement intention. The results support the feasibility of future rehabilitative treatments based on the combination of robot-assisted physical therapy with BCIs.