Robot-assisted gait training in multiple sclerosis patients: a randomized trial.

BACKGROUND: Preservation of locomotor activity in multiple sclerosis (MS) patients is of utmost importance. Robotic-assisted body weight-supported treadmill training is a promising method to improve gait functions in neurologically impaired patients, although its effectiveness in MS patients is still unknown. OBJECTIVE: To compare the effectiveness of robot-assisted gait training (RAGT) with that of conventional walking treatment (CWT) on gait and generalized functions in a group of stable MS patients. METHODS: A prospective randomized controlled trial of 12 sessions of RAGT or CWT in MS patients of EDSS score 5-7. Primary outcome measures were gait parameters and the secondary outcomes were functional and quality of life parameters. All tests were performed at baseline, 3 and 6 months post-treatment by a blinded rater. RESULTS: Fifteen and 17 patients were randomly allocated to RAGT and CWT, respectively. Both groups were comparable at baseline in all parameters. As compared with baseline, although some gait parameters improved significantly following the treatment at each time point there was no difference between the groups. Both FIM and EDSS scores improved significantly post-treatment with no difference between the groups. At 6 months, most gait and functional parameters had returned to baseline. CONCLUSIONS: Robot-assisted gait training is feasible and safe and may be an effective additional therapeutic option in MS patients with severe walking disabilities.

The effectiveness of locomotor therapy using robotic-assisted gait training in subacute stroke patients: a randomized controlled trial.

OBJECTIVE: To evaluate the effectiveness of early and prolonged locomotor treatment with the use of a robotic-assisted gait training (RAGT) device (Lokomat; Hocoma Inc., Zurich, Switzerland) on the functional outcomes of patients after subacute stroke. DESIGN: A nonblinded prospective, randomized, controlled study. SETTING: Rehabilitation department in tertiary university medical center. PATIENTS: Sixty-seven patients in the first 3 months after subacute stroke were randomized into 2 groups as follows. Thirty-seven patients were treated with RAGT, and 30 were treated with regular physiotherapy. Inclusion criteria were first stroke, independent ambulation before the stroke, and neurological severity between 6 and 20 according to the National Institutes of Health Stroke Scale (NIHSS). INTERVENTION: RAGT treatment was administered 3 times a week for 30 minutes, combined with regular physiotherapy for 6 weeks. Control patients received the equivalent additional time of regular physiotherapy. MAIN OUTCOME MEASUREMENTS: The primary outcome was the ability to walk independently, as assessed by use of the functional ambulatory capacity scale. The secondary outcomes included the neurological status according to the NIHSS; functional motor assessment (determined by use of the stroke activity scale); and gait parameters, including gait velocity, endurance, and number of climbed stairs. RESULTS: In the intention-to-treat analysis, subjects in the RAGT group exhibited greater gains than the control group in their ability to walk independently, as expressed by a greater functional ambulatory capacity score (P < .01), and in their neurological status according to NIHSS (P < .01). Among those who achieved independent walking, nonsignificant differences between groups were noted according to secondary outcome measures of gait parameters except from step climbing. CONCLUSION: This controlled study showed, at the end of a 6-week trial, that locomotor therapy with the use of RAGT combined with regular physiotherapy produced promising effects on functional and motor outcomes in patients after subacute stroke as compared with regular physiotherapy alone.

Spinal decompression sickness presenting as partial Brown-Sequard syndrome and treated with robotic-assisted body-weight support treadmill training.

OBJECTIVE: To describe the rehabilitation outcome of a case of spinal decompression sickness presenting as partial Brown-Sequard syndrome treated with robotic-assisted body-weight support treadmill training. STUDY DESIGN: Case report. BACKGROUND: Type II decompression sickness patients commonly suffer from myelopathy with gait disturbances necessitating rehabilitation. Robotic-assisted body-weight support treadmill training has been shown to improve the rehabilitation outcome of incomplete spinal cord injury. Its usefulness has not been described in decompression sickness myelopathy. METHODS: Robotic-assisted body-weight support treadmill training was administrated using the Lokomat. Primary outcomes were American Spinal Cord Association scale, Spinal Cord Independence Measurement, Berg Balance Test, and Walking Index for Spinal Cord Injury. RESULTS: The patient was admitted 3 weeks after the diving injury, with severe paraparesis and a T11 sensory neurological level, resembling partial Brown-Sequard syndrome. After 3 months of rehabilitation including 18 Lokomat sessions, American Spinal Cord Association score improved from C to D, Spinal Cord Independence Measurement improved from 50 to 90 out of 100. Berg Balance Test improved from 35 to 43 out of 56 and Walking Index for Spinal Cord Injury improved from 1 to 15 out of 20. Upon discharge he could walk with one crutch for more than 1 km. CONCLUSION: Robotic-assisted body-weight support treadmill training for spinal decompression sickness rehabilitation might be beneficial.

Reliability and validity of the modified functional reach test at the sub-acute stage post-stroke.

OBJECTIVES: The first aim of this study was to evaluate the within-session reliability of sitting balance measures by assessing forward and lateral reach while sitting in both healthy subjects and patients post- stroke. The second aim was to evaluate the ability to document change in reaching while sitting over time in patients post-stroke. The third aim was to compare sitting balance results by the modified functional reach test (MFRT) to the Balance Master (BM), motor and function assessments. DESIGN: Data were collected on two occasions: Two to three weeks post-event and again six weeks later. On each occasion within-session reliability was tested using the intra-class correlations (ICC). The tests were performed three times; the second and third attempts were compared in order to test the within-session reliability. For assessing the concurrent validity, the MFRT results were compared with the BM results, Stroke Assessment Scale (SAS) and the Functional Independence Measure (FIM) score using Pearson correlations. SETTING: In-patient rehabilitation department. PARTICIPANTS: Patients after a first ischemic stroke 14 - 21 days post-event were recruited from the inpatient rehabilitation department at Hadassah University Hospital, Jerusalem, Israel. Excluded were patients with brain stem lesions and/or bilateral signs or hemorrhagic events as diagnosed by Computerized Tomography, patients with Mini-Mental State Examination < 20, those who were not able to sit unsupported for 10 sec and those who could stand without support for more than 30 sec. MAIN OUTCOME MEASURES: The MFRT, performed while sitting in forward and sideward directions. RESULTS: The MFRT in all directions on both occasions exhibited high reliability (intra-class correlation coefficient range, 0.90 - 0.97). The responsiveness to the paretic side was high (effect size 0.80) and moderate for the forward and non-paretic side (effect size 0.57 - 0.60). A significant moderate correlation was found between MFRT and BM on both occasions. CONCLUSION: The MFRT while sitting can be reliably measured and may serve as a useful outcome measure in individuals with stroke 2 - 8 weeks post-event.

beta-Carotene interferes with ultraviolet light A-induced gene expression by multiple pathways.

Ultraviolet light A (UVA) exposure is thought to cause skin aging mainly by singlet oxygen ((1)O(2))-dependent pathways. Using microarrays, we assessed whether pre-treatment with the (1)O(2) quencher beta-carotene (betaC; 1.5 microM) prevents UVA-induced gene regulation in HaCaT human keratinocytes. Downregulation of growth factor signaling, moderate induction of proinflammatory genes, upregulation of immediate early genes including apoptotic regulators and suppression of cell cycle genes were hallmarks of the UVA effect. Of the 568 UVA-regulated genes, betaC reduced the UVA effect for 143, enhanced it for 180, and did not interact with UVA for 245 genes. The different interaction modes imply that betaC/UVA interaction involved multiple mechanisms. In unirradiated keratinocytes, gene regulations suggest that betaC reduced stress signals and extracellular matrix (ECM) degradation, and promoted keratinocyte differentiation. In irradiated cells, expression profiles indicate that betaC inhibited UVA-induced ECM degradation, and enhanced UVA induction of tanning-associated protease-activated receptor 2. Combination of betaC-promoted keratinocyte differentiation with the cellular "UV response" caused synergistic induction of cell cycle arrest and apoptosis. In conclusion, betaC at physiological concentrations interacted with UVA effects in keratinocytes by mechanisms that included, but were not restricted to (1)O(2) quenching. The retinoid effect of betaC was minor, indicating that the betaC effects reported here were predominantly mediated through vitamin A-independent pathways.

Expression modes of interferon-alpha inducible genes in sensitive and resistant human melanoma cells stimulated with regular and pegylated interferon-alpha.

Interferon-alpha with its antiproliferative activity is widely used for the treatment of viral infections and tumor therapy such as melanoma. Naturally occurring resistance to recombinant interferon alpha-2a (IFN-alpha) and severe side effects limit the therapeutic efficacy. Understanding of the molecular mechanisms involved in unresponsiveness may therefore lead to the development of novel formulations that overcome resistance. Here, we have applied oligonucleotide DNA microarrays with probe sets for about 11,400 human transcripts to study the expression of interferon-alpha inducible genes in a sensitive and resistant melanoma cell line over multiple time points and two interferon formulations. We identified two major groups of genes with termed interferon primary response genes (IPRGs) or interferon secondary response genes (ISRGs). IPRGs are upregulated early after interferon stimulation in both the sensitive and the resistant line and they contain IREs in the noncoding regulatory region. In contrast, ISRG expression occurs preferentially in the sensitive line ME15 at late time points, and this group of genes lacks typically IREs. In addition to these two major interferon response gene classes, we identified a relatively small number of genes with complex kinetic expression modes. In addition, we show for the first time that regular and pegylated recombinant interferons are equally potent activators of interferon (IFN) gene expression. Finally, we propose that the ISRGs are activated downstream of the primary response genes by a molecule or pathway, which awaits identification, and interferon inducible gene expression is thus more complicated than previously thought.

Statistical methods for analyzing microarray feature data with replications.

Expression levels in oligonucleotide microarray experiments depend on a potentially large number of factors, for example, treatment conditions, different probes, different arrays, and so on. To dissect the effects of these factors on expression levels, fixed-effects ANOVA methods have previously been proposed. Because we are not necessarily interested in estimating the specific effects of different probes and arrays, we propose to treat these as random effects. Then we only need to estimate their means and variances but not the effect of each of their levels; that is, we can work with a much reduced number of parameters and, consequently, higher precision for estimating expression levels. Thus, we developed a mixed-effects ANOVA model with some random and some fixed effects. It automatically accounts for local normalization between different arrays and for background correction. The method was applied to each of the 6,584 genes investigated in a microarray experiment on two mouse cell lines, PA6/S and PA6/8, where PA6/S enhances proliferation of Pre B cells in vitro but PA6/8 does not. To detect a set of differentially expressed genes (multiple testing problem), we applied the method of controlling the false discovery rate (FDR), which successfully identified 207 genes with significantly different expression levels.

Analysis of the eye developmental pathway in Drosophila using DNA microarrays.

Pax-6 genes encode evolutionarily conserved transcription factors capable of activating the gene-expression program required to build an eye. When ectopically expressed in Drosophila imaginal discs, Pax-6 genes induce the eye formation on the corresponding appendages of the adult fly. We used two different Drosophila full-genome DNA microarrays to compare gene expression in wild-type leg discs versus leg discs where eyeless, one of the two Drosophila Pax-6 genes, was ectopically expressed. We validated these data by analyzing the endogenous expression of selected genes in eye discs and identified 371 genes that are expressed in the eye imaginal discs and up-regulated when an eye morphogenetic field is ectopically induced in the leg discs. These genes mainly encode transcription factors involved in photoreceptor specification, signal transducers, cell adhesion molecules, and proteins involved in cell division. As expected, genes already known to act downstream of eyeless during eye development were identified, together with a group of genes that were not yet associated with eye formation.

Changes in gene expression profiles in developing B cells of murine bone marrow.

Gene expression profiles of five consecutive stages of mouse B cell development were generated with high-density oligonucleotide arrays from as few as 2 x 10(4) ex vivo isolated and flow-cytometrically purified cells. Between 2.8% and 6.8% of all genes change on differentiation from one cellular stage to the next by at least twofold. The entire pathway involves differential expression of 10.7% of all genes. Previously known expression patterns of 15 genes (like surrogate light chain, RAG-1/2, MHC class II, mel-14 antigen) are confirmed. The gene expression patterns of the proliferating pre-BI and large pre-BII cells on the one hand, and the resting immature and mature B cells on the other hand, are most similar to each other. Small pre-BII cells display a pattern that is transitional between these two groups. Most of the genes expressed in early precursors are involved in general processes, like protein folding or cell cycle regulation, whereas more mature precursors express genes involved in more specific molecular programs (cell surface receptors, secreted factors, and adhesion molecules, among others). Between 19 and 139 genes share a given expression pattern. Combining knowledge about gene function and expression pattern allows identification of novel candidate genes potentially involved in self-maintenance of pre-BI cells, allelic exclusion and pre-B cell receptor signaling in large pre BII cells, cell-cycle arrest of small pre-BII cells, propensity toward apoptosis or anergization in immature B cells, propensity toward cell division and activation in mature B cells, and stage-specific interactions with stromal cells in the bone marrow.