Backbone 1H, 13C and 15N resonance assignment of the ubiquitin specific protease 7 catalytic domain (residues 208-554) in complex with a small molecule ligand.

The backbone 1H, 13C and 15N resonance assignment of Ubiquitin Specific Protease 7 catalytic domain (residues 208-554) was performed in its complex with a small molecule ligand and in its apo form as a reference. The amide 1H-15N signal intensities were boosted by an amide hydrogen exchange protocol, where expressed 2H, 13C, 15N-labeled protein was unfolded and re-folded to ensure exchange of amide deuterons to protons. The resonance assignments were used to determine chemical shift perturbations on ligand binding, which are consistent with the binding site observed by crystallography.

Patients' experiences of unilateral spatial neglect between stroke onset and discharge from inpatient rehabilitation: a thematic analysis of qualitative interviews.

Purpose: The purpose of this qualitative study was to explore how stroke patients with unilateral spatial neglect experience; (a) performance in activities of daily living; (b) alterations in bodily perceptions; and (c) personal hopes and expectations, looking at the period between stroke onset and discharge from inpatient rehabilitation.Materials and methods: We conducted individual semi-structured interviews with 7 (5 men, 2 women, mean age 69 years) consecutively sampled participants. All interviews were transcribed verbatim and analyzed using thematic analysis.Findings: Participants' experiences were captured in three over-arching themes: "unawareness of neglect," "emergent awareness for neglect-related difficulties," and "comparing the new life with the old one." Findings showed that participants progressed from initial unawareness to emergent awareness for their neglect-related difficulties over the course of rehabilitation. Comparing their current life situation with the one before their stroke triggered feelings of uncertainty and regret, with associated decreased pleasure in meaningful activities.Conclusions: This study informs health professionals regarding personal experiences of orientation in and reorganization of life of stroke patients with unilateral spatial neglect. The findings highlight that being aware of neglect-related deficits is a prerequisite for using coping strategies and incorporating them in daily life. Possible therapeutic strategies that fit the current stage of recovery and level of awareness are discussed.IMPLICATIONS FOR REHABILITATIONSymptoms of unilateral spatial neglect and anosognosia are most common after right hemispheric stroke, both being predictors of poorer functional outcome during rehabilitation.Patients' regaining some awareness of their impairments is a prerequisite for successful treatment and for engagement in neglect-specific interventions.Health care providers should carefully examine the "state of awareness" of the patient and adapt their therapeutic approach accordingly.This check should frequently be repeated over the course of rehabilitation, as awareness changes at different paces for each patient.

GPR103 antagonists demonstrating anorexigenic activity in vivo: design and development of pyrrolo[2,3-c]pyridines that mimic the C-terminal Arg-Phe motif of QRFP26.

GPR103, a G-protein coupled receptor, has been reported to have orexigenic properties through activation by the endogenous neuropeptide ligands QRFP26 and QRFP43. Recognizing that central administration of QRFP26 and QRFP43 increases high fat food intake in rats, we decided to investigate if antagonists of GPR103 could play a role in managing feeding behaviors. Here we present the development of a new series of pyrrolo[2,3-c]pyridines as GPR103 small molecule antagonists with GPR103 affinity, drug metabolism and pharmacokinetics and safety parameters suitable for drug development. In a preclinical obesity model measuring food intake, the anorexigenic effect of a pyrrolo[2,3-c]pyridine GPR103 antagonist was demonstrated. In addition, the dynamic 3D solution structure of the C-terminal heptapeptide of the endogenous agonist QRFP26(20-26) was determined using NMR. The synthetic pyrrolo[2,3-c]pyridine antagonists were compared to this experimental structure, which displayed a possible overlay of pharmacophore features supportive for further design of GPR103 antagonists.

The FeSTivaLS trial protocol: a randomized evaluation of the efficacy of functional strength training on enhancing walking and upper limb function later post stroke.

RATIONALE: Functional Strength Training may enhance motor function of people who are more than six months post stroke. AIMS: to evaluate the clinical efficacy of enhancing upper and lower limb motor function with FST to explore participants' views (expectations and experiences) of FST, and to determine what cost-effectiveness data to collect in a subsequent Phase III trial. DESIGN: Randomized, observer-blind trial with embedded qualitative investigation of participants' views of FST (n = 6, purposive sampling). STUDY: Participants (n = 58), six months to five years after stroke with difficulty using their paretic upper (UL) and lower limbs (LL) for everyday functional activity. All will be randomized to either FST-UL or FST-LL delivered in their own homes for four days each week for six weeks. FST involves repetitive progressive resisted exercise during goal directed functional activities. The therapist's main input is to provide verbal prompting and feedback. OUTCOMES: Measures will be undertaken before randomization (baseline), after the six-week intervention (outcome) and six weeks thereafter (follow-up). Primary outcomes for clinical efficacy will be the Functional Ambulation Categories (FAC) and the Action Research Arm Test (ARAT). Clinical efficacy analysis will use the proportional odds model for FAC and a Mann-Whitney test for ARAT. Participants' views of FST will be explored at baseline and outcome through audiotaped, semi-structured, narrative approach, interviews. The analytic process for interviews will sort transcribed data thematically and seek categories to inform conceptualization (theory-building). A purpose-designed cost questionnaire will identify what cost resource items are likely to be affected by FST.

An evaluation of the effects of a dynamic lycra orthosis on arm function in a late stage patient with acquired brain injury.

PRIMARY OBJECTIVE: The aim of this study was to assess the effect of a dynamic lycra orthosis in the management of upper limb paresis in a late stage stroke patient. RESEARCH DESIGN: A single case experiment, adopting a 3-phase ABA withdrawal design (without follow-up), approximately 6-weeks per phase, intervention being delivered in the middle/B phase. METHOD AND PROCEDURES: Assessment of arm function was carried out on a weekly basis for the 18-weeks, using a battery of upper limb tests. The subject was prescribed a tailor-made lycra orthosis which was worn daily during the middle phase of the trial. MAIN OUTCOMES AND RESULTS: Orthosis wear appeared to result in improvements in active range of movement, self-rated function and one component of a writing test, with some suggestion of a carryover effect when treatment was withdrawn. No intervention-related changes were seen in a dressing test. Ambiguous results were seen in a further writing test and a peg board manipulation assessment. Irrespective of intervention, the subject made positive changes in scores in the majority of assessments used, throughout the 18-week period. CONCLUSIONS: The findings suggest that a lycra orthosis may have some beneficial effects on upper limb function late after brain injury. Results were however equivocal, suggesting (a) that effect mechanisms may be quite complex and (b) that future evaluations may require careful construction.

Path tree analysis as a tool for describing motor recovery following acquired brain injury: an exploratory study.

PURPOSE: Knowledge of the likely sequence of motor recovery following brain injury has obvious usefulness within rehabilitation. Research has tended to focus on establishing the linearity of the recovery sequence. This study aimed to ascertain the usefulness of path tree analysis as a graphical means of exploring inter-subject variation in routes followed during motor recovery. METHOD: Data were collected via physiotherapist questionnaire, ascertaining recovery sequence of 14 motor milestones across 80 adults with severe traumatic brain injury (TBI). An adapted version of the Rivermead Mobility Index (RMI) was used. Data were analysed to establish inter-subject agreement regarding recovery sequence. A path tree diagram was then composed to explore inter-subject variation. RESULTS: Putative order of recovery for the 14 motor milestones appeared to have face validity. Guttman coefficients of reproducibility ((0.954) and scalability (0.948) were good, suggesting intrinsic order. Composition of a path tree suggested considerable variation in recovery paths between individuals, with 34 combinations of test items being evident. CONCLUSIONS: Motor recovery following severe TBI follows a distinct pattern, but path tree analysis emphasises the variety of routes which individuals can follow. Additional research should ascertain the scope of this method to document the natural history of the recovery process.

Determinants of the endosomal localization of sorting nexin 1.

The sorting nexin (SNX) family of proteins is characterized by sequence-related phox homology (PX) domains. A minority of PX domains bind with high affinity to phosphatidylinositol 3-phosphate [PI(3)P], whereas the majority of PX domains exhibit low affinity that is insufficient to target them to vesicles. SNX1 is located on endosomes, but its low affinity PX domain fails to localize in vivo. The NMR structure of the PX domain of SNX1 reveals an overall fold that is similar to high-affinity PX domains. However, the phosphatidylinositol (PI) binding pocket of the SNX1 PX domain is incomplete; regions of the pocket that are well defined in high-affinity PX domains are highly mobile in SNX1. Some of this mobility is lost upon binding PI(3)P. The C-terminal domain of SNX1 is a long helical dimer that localizes to vesicles but not to the early endosome antigen-1-containing vesicles where endogenous SNX1 resides. Thus, the obligate dimerization of SNX1 that is driven by the C-terminal domain creates a high-affinity PI binding species that properly targets the holo protein to endosomes.