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Biomedical subjects

J Johnston

Publications and source records attributed to J Johnston.

At least 19 recordsLinked to original sources

A study of linkage and association of body mass index in the Old Order Amish.

Obesity is thought to have a genetic component with the estimates of heritability ranging from 0.25-0.40. As part of an ongoing study of obesity in the Old Order Amish, seven two- and three-generation families (157 individuals) were assessed for 21 traits related to obesity, including body mass index (BMI) and BMI-percentile (a standardized distribution of BMI adjusted for age and sex). Genotyping was performed using a panel of 384 short-tandem repeat markers. In this sample, the estimates of heritability ranged from 0.16-0.31 for BMI and from 0.40-0.52 for BMI-percentile. Model-independent linkage analysis identified candidate regions on chromosomes 1, 5, 7, 8, and 11. Given that several markers on 7q were significant for both BMI and BMI-percentile (P < or = 0.001) and that the structural locus for leptin was located on 7q, this region was considered to be the primary candidate region. Subsequent typing of additional flanking markers on 7q corroborated the original findings. Tests of intrafamilial association for alleles at markers in this candidate region were significant at similar levels. Although there is some evidence for linkage and association in the region containing leptin, there appears to be stronger evidence for linkage (P < or = 0.001) and association (P < or = 0.00001) with BMI in a region 10-15 cM further downstream of leptin, flanked by markers D7S1804 and D7S3070 with peak values from D7S495-D7S1798. Evidence from linkage and association studies suggests that this region (D7S1804-D7S3070) may be responsible, at least in part, for variation in BMI and BMI-percentile in the Old Order Amish.

Alleles↗

The role of sub-maximal force production in the enslaving phenomenon.

When individuals perform a force task involving only one finger, they involuntarily move other fingers as well. This phenomenon is referred to as the enslaving force or the interdependency of fingers. Given that previous literature on the enslaving force has focused on maximal isometric force production, the present research was designed to study the role of sub-maximal force production in the enslaving phenomenon. To this end, we examined behaviorally three levels of force production with a constant rate of force development. We also examined the temporal organization of enslaving separating the achievement of the desired force (ramp phase) and its maintenance (static phase). During the static phase we found: (i) the amount of enslaving increased with the increment of nominal force level whether the index, middle, ring or little fingers were used as the master finger; (ii) enslaving is strongest in the finger directly adjacent to the master finger; and (iii) in terms of enslaving, the index finger was more 'independent' than the other three fingers, regardless of nominal force produced, followed by the little, middle, and ring fingers. In terms of temporal organization, we found that the time-lag of activation of 'slave fingers' during the ramp phase was reduced as the amount of force level increased. Overall, our data suggest that enslaving effect is a task specific phenomenon and depends on the amount of force produced by the master finger.

Adult↗

Movement-related EEG potentials are force or end-effector dependent: evidence from a multi-finger experiment.

OBJECTIVES: This study examined behavioral and electrocortical responses in producing 3 levels of force (25, 50 and 75% of MVC) at a constant rate of force development with each of 4 fingers both during the achievement of the desired force (ramp phase) and its maintenance (static phase). We were particularly interested in describing in more detail the interaction between nominal force and finger on various components of movement-related potential (MRP) associated with preparation and execution of isometric force production tasks. METHODS: Our experimental design systematically controlled the rate of force development while nominal force level was experimentally manipulated during isometric force production tasks. We applied time-domain averaging of EEG single trials in order to extract 3 components of MRP (BP(-600 to -500); MP(-100 to 0); MMP) preceding and accompanying behavioral responses. RESULTS: Overall, as in our previous research the effect of force per se was not reflected in the EEG components. However, we did find an interaction between finger and force level in both the Bereitshaftspotential (BP) and motor potential (MP) components of the movement-related potentials. While the middle, ring and little finger produced no differences in EEG components at any of the 3 force levels, the index finger did. We further correlated the force trajectory and the EEG time series with the highest correlations found in the lowest force level with the index finger. As the force level was increased, the correlation was significantly reduced. CONCLUSIONS: Overall, the whole complex of MRP components and evolution of EEG time series during multi-finger isometric force production tasks reflect a combination of factors including the primary end-effector performing the task and interaction of end-effector and the amount of nominal force.

Adult↗

Motor-related cortical potentials accompanying enslaving effect in single versus combination of fingers force production tasks.

OBJECTIVES: This study examined behavioral indices and motor-related cortical potentials (MRCP) of the enslaving phenomenon (i.e. interdependency of finger movement) during isometric force production tasks using each of the four fingers separately and in combination. We examined MRCP preceding force production and those during the achievement of the desired force (ramp phase) and its maintenance (static phase). METHODS: Our experimental design systematically controlled the isometric force output, including both ramp and static phases of force production. We applied time-domain averaging of electroencephalographic single trials in order to extract 3 components of MRCP (Bereitshaftspotential, motor potentials, and motor monitoring potentials) preceding and accompanying force responses. RESULTS: We report two major findings. First, we found the index finger to be more independent, accurate, and to display the larger MRCP amplitude whereas the ring finger was more dependent, less accurate, and displayed smaller MRCP amplitude. Second, adding the neighboring finger when the ring finger produced the task significantly reduced its dependency on uninvolved fingers and increased the accuracy of both ramp and static phases which was not the case with the index finger. The amplitude of MRCP was increased when the ring finger produced the task in combination as compared to when the ring finger performed the task in isolation. In contrast, the amplitude of MRCP was significantly reduced when the index finger produced the task in combination with other fingers when compared to when the index finger performed the task in isolation. CONCLUSIONS: Overall, the amount of the fingers' dependency on the uninvolved fingers (e.g. amount of enslaving) during isometric force production tasks was inversely related with the amplitude of MRCP indicating the contribution of central mechanisms to the enslaving phenomenon.

Adult↗

Modulated cortical control of individual fingers in experienced musicians: an EEG study. Electroencephalographic study.

OBJECTIVE: The present research was designed to address the nature of interdependency between fingers during force production tasks in subjects with varying experience in performing independent finger manipulation. Specifically, behavioral and electroencephalographic (EEG) measures associated with controllability of the most enslaved (ring) and the least enslaved (index) fingers was examined in musicians and non-musicians. METHODS: Six piano players and 6 age-matched control subjects performed a series of isometric force production tasks with the index and ring fingers. Subjects produced 3 different force levels with either their index or ring fingers. We measured the isometric force output produced by all 4 fingers (index, ring, middle and little), including both ramp and static phases of force production. We applied time-domain averaging of EEG single trials in order to extract 4 components of the movement-related cortical potentials (MRCP) preceding and accompanying force responses. RESULTS: Three behavioral findings were observed. First, musicians were more accurate than non-musicians at reaching the desired force level. Second, musicians showed less enslaving as compared to non-musicians. And third, the amount of enslaving increased with the increment of nominal force levels regardless of whether the index or ring finger was used as the master finger. In terms of EEG measures, we found differences between tasks performed with the index and ring fingers in non-musicians. For musicians, we found larger MRCP amplitudes at most electrode sites for the ring finger. CONCLUSIONS: Our data extends previous enslaving research and suggest an important role for previous experience in terms of the independent use of the fingers. Given that a variety of previous work has shown finger independence to be reflected in cortical representation in the brain and our findings of MRCP amplitude associated with greater independence of fingers in musicians, this suggests that what has been considered to be stable constraints in terms of finger movements can be modulated by experience. SIGNIFICANCE: This work supports the idea that experience is associated with changes in behavioral and EEG correlates of task performance and may have clinical implications in disorders such as stroke or focal hand dystonia. Practice-related procedures offer useful approaches to rehabilitation strategies.

Adult↗

Human thymic stromal lymphopoietin preferentially stimulates myeloid cells.

The sequence of a novel hemopoietic cytokine was discovered in a computational screen of genomic databases, and its homology to mouse thymic stromal lymphopoietin (TSLP) suggests that it is the human orthologue. Human TSLP is proposed to signal through a heterodimeric receptor complex that consists of a new member of the hemopoietin family termed human TSLP receptor and the IL-7R alpha-chain. Cells transfected with both receptor subunits proliferated in response to purified, recombinant human TSLP, with induced phosphorylation of Stat3 and Stat5. Human TSLPR and IL-7Ralpha are principally coexpressed on monocytes and dendritic cell populations and to a much lesser extent on various lymphoid cells. In accord, we find that human TSLP functions mainly on myeloid cells; it induces the release of T cell-attracting chemokines from monocytes and, in particular, enhances the maturation of CD11c(+) dendritic cells, as evidenced by the strong induction of the costimulatory molecules CD40 and CD80 and the enhanced capacity to elicit proliferation of naive T cells.

Amino Acid Sequence↗

Barth's syndrome-like disorder: a new phenotype with a maternally inherited A3243G substitution of mitochondrial DNA (MELAS mutation).

An Argentine male child died at 4.5 years of age of a lethal mitochondrial disease associated with a MELAS mutation and a Barth syndrome-like presentation. The child had severe failure to thrive from the early months and for approximately two years thereafter. In addition, the patient had severely delayed gross motor milestones, marked muscle weakness, and dilated cardiomyopathy that progressed to congestive heart failure. He also had persistently elevated urinary levels of 3-methylglutaconic and 2-ethylhydracrylic acids and low blood levels of cholesterol. Detailed histopathologic evaluation of the skeletal muscle biopsy showed high activity of succinate dehydrogenase, a generalized decrease of COX activity, and abundant ragged-red fibers. Electron microscopic studies revealed multiple mitochondrial abnormalities in lymphocytes and monocytes, in the striated muscle, and in the postmortem samples (muscle, heart, liver, and brain). Biochemical analysis showed a pronounced and constant lactic acidosis, and abnormal urinary organic acid excretion (unchanged in the fasting and postprandial states). In addition, in CSF there was a marked increase of lactate and beta-hydroxybutyrate (beta-HOB) and also a high systemic ratio beta-HOB/acetoacetate. Enzymatic assay of the respiratory chain in biopsied muscle showed 10% of complex I activity and 24% of complex IV activity compared with controls. Molecular studies of the mitochondrial genome revealed an A to G mutation at nucleotide pair 3243 in mitochondrial DNA, a well-known pathogenetic mutation (MELAS mutation) in all the patient's tissues and also in the blood specimens of the probands mother and sibs (4 of 5). The diagnosis of MELAS mutation was reinforced by the absence of an identifiable mutation in the X-linked G4.5 gene of the propositus. The present observation gives additional evidence of the variable clinical expression of mtDNA mutations in humans and demonstrates that all clinical variants deserve adequate investigation to establish a primary defect. It also suggests adding Barth-like syndrome to the list of phenotypes with the MELAS mutation.

3-Hydroxybutyric Acid↗

Morbidity and mortality in rheumatoid arthritis patients with prolonged and profound therapy-induced lymphopenia.

OBJECTIVE: Therapies that deplete lymphocytes often improve symptoms in patients with otherwise refractory autoimmune disease but may result in long-term lymphopenia, the consequences of which are uncertain. To assess the impact of prolonged lymphopenia on morbidity and mortality, we studied patients who had previously received lymphocytotoxic monoclonal antibody (mAb) therapy for rheumatoid arthritis (RA). METHODS: Fifty-three patients who received the lymphocytotoxic mAb CAMPATH-1H between 1991 and 1994 in the United Kingdom were assessed for mortality and infectious and malignant morbidity, by interview and case-note review. In addition, patients were monitored via the National Health Service Central Registry, to verify notification of death. Peripheral blood lymphocyte subsets were analyzed by flow cytometry. A retrospective, matched-cohort study of mortality was also performed with 102 control subjects selected from the European League Against Rheumatism database, which comprises patients with rheumatic disorders who have received immunosuppressive drugs. RESULTS: There was profound and persistent peripheral blood lymphopenia in the mAb-treated patients, affecting predominantly the CD4+ subset. Median CD4+, CD8+, and CD19+ peripheral blood lymphocyte counts at 73-84 months after therapy were 185 cells/microl, 95 cells/microl, and 115 cells/microl, respectively. At a median followup of 71 months (range 14-90), 13 patients had died (24.5%), compared with 18% of the matched controls, providing a mortality rate ratio of 1.45 (95% confidence interval 0.65-3.13). During 283 patient-years of followup, there were 36 infections classified as major (12.7 per 100 patient-years). The causes of death and the spectrum of infections documented were similar to those expected in a hospital-based RA cohort. Patients who received more than 1 course of therapy had more severe lymphopenia than did patients who received a single course, but this did not have an impact on mortality or morbidity. CONCLUSION: Despite the occurrence of profound and long-lasting lymphopenia following treatment with antilymphocyte mAb therapy for RA, this therapy is not associated with a large excess of mortality nor with an unusual spectrum of infections, at least during a medium-term period of followup. These data are also relevant to patients receiving lymphocytotoxic mAb therapy for other indications, and to patients receiving other lymphodepleting therapies such as autologous stem cell transplantation.

Aged↗

Sequestration of adenoviral vector by Kupffer cells leads to a nonlinear dose response of transduction in liver.

Systemic administration of a recombinant adenovirus encoding the human interferon-beta gene (H5.110CMVhIFN-beta) results in transduction of hepatocytes and detectable circulating levels of IFN-beta protein. In preclinical studies in mice, we noticed a distinctly nonlinear dose response, with low levels of virus (1-3 x 10(10) viral particles) yielding barely detectable levels of IFN-beta but with a higher viral dose (1 x 10(11) particles) resulting in disproportionately high IFN-beta levels. Further studies showed that transgene expression levels from low viral doses could be dramatically enhanced by coadministering an unrelated recombinant adenovirus (H5.110CMVlacZ), suggesting that there was a viral dose threshold effect for efficient viral transduction and/or IFN-beta expression. This enhancement of reporter expression by a nonreporter adenovirus, effective upon coadministration, was further enhanced by preadministration of H5.110CMVlacZ (up to 8 h), but was ineffective if the helper virus was administered as little as 5 min after the H5.110CMVhIFN-beta reporter virus. Our data suggest that the reticuloendothelial system plays a role in this threshold effect, such that low doses of virus are efficiently taken up by the RES/Kupffer cells without leading to appreciable transgene expression, whereas high doses saturate these cells and are able to productively transduce hepatocytes. A better understanding of this phenomenon could have an impact on gene therapy clinical trial safety and efficacy.

Adenoviridae↗

TACI-Ig neutralizes molecules critical for B cell development and autoimmune disease. impaired B cell maturation in mice lacking BLyS.

BLyS and APRIL have similar but distinct biological roles, mediated through two known TNF receptor family members, TACI and BCMA. We show that mice treated with TACI-Ig and TACI-Ig transgenic mice have fewer transitional T2 and mature B cells and reduced levels of circulating immunoglobulin. TACI-Ig treatment inhibits both the production of collagen-specific Abs and the progression of disease in a mouse model of rheumatoid arthritis. In BLyS-deficient mice, B cell development is blocked at the transitional T1 stage such that virtually no mature B cells are present, while B-1 cell numbers are relatively normal. These findings further elucidate the roles of BLyS and APRIL in modulating B cell development and suggest that BLyS is required for the development of most but not all mature B cell populations found in the periphery.

Animals↗

Movement-related cortical potentials associated with progressive muscle fatigue in a grasping task.

OBJECTIVE: The present research was aimed to further address the general empirical question regarding the behavioral and neurophysiological indices and mechanisms that contribute to and/or compensate for muscle fatigue. In particular, we examined isometric force production, EMG, and EEG correlates of progressive muscle fatigue while subjects performed a grasping task. METHODS: Six neurologically healthy subjects were instructed to produce and maintain 70% of maximum voluntary contraction (MVC) for a total of 5 s in a sequence of 120 trials using a specially designed grip dynamometer. Three components of movement-related potentials (Bereitschaftspotential, BP, Motor potential, MP, and Movement-monitoring potential, MMP) were extracted from continuous EEG records and analyzed with reference to behavioral indicators of muscle fatigue. RESULTS: Experimental manipulations induced muscle fatigue that was demonstrated by decreases in both MVC values and mean force levels produced concomitant to increases in EMG root mean square (RMS) amplitude with respect to baseline levels, and EMG slope. EEG data revealed a significant increase in MP amplitude at precentral (Cz and FCz) and contralateral (C3) electrode sites, and increases in BP amplitude at precentral (Cz and FCz) electrode sites. CONCLUSIONS: The increases in EMG amplitude, EMG slope, and MP amplitudes suggest a possible link between the control signal originating in the motor cortex and activity level of the alpha-motoneuron pool as a function of progressive muscle fatigue. Overall, the data demonstrate that progressive muscle fatigue induced a systematic increase in the electrocortical activation over the supplementary motor and contralateral sensorimotor areas as reflected in the amplitude of movement-related EEG potentials.

Adolescent↗

Immediate-early protooncogene expression and liver function following various extents of partial hepatectomy in the rat.

Immediate-early protooncogenes (IEP) are thought to play an important role in hepatocyte replication. Whether the extent of their expression correlates with the strength of the proliferative stimulus and subsequent regenerative activity has yet to be documented in vivo. Data are also lacking with respect to the level at which liver disease is associated with biochemical evidence of hepatic dysfunction. Thus, the objectives of this study were to determine whether a correlation exists between IEP gene mRNA expression and varying extents of partial hepatectomy (PHx) and to document the extent of resection required to result in increases in serum bilirubin levels. Eighty-nine adult, male Sprague-Dawley rats underwent either sham surgery or 20%, 35%, 55%, 70% or 90% PHx. Postoperatively, rats were killed (N = 3-6/group) at 15 and 30 mins and 8 and 24 hrs for c-fos, c-jun, and c-myc mRNA expression by northern blot analyses. Rats killed at 24 hrs also had hepatic regenerative activity documented by [3H]thymidine incorporation into hepatic DNA and serum bilirubin determinations. While c-fos mRNA expression at 15 mins and c-myc mRNA expression at 8 hrs after PHx did not correlate with the extent of PHx (r2 = 0.478 and 0.018, respectively), a weak correlation existed between c-jun mRNA expression at 30 mins and the extent of PHx (r2 = 0.662, P < 0.05). In terms of IEP mRNA expression and hepatic regenerative activity, a strong correlation existed between c-fos mRNA expression and [3H]thymidine incorporation (r2 = 0.851, P < 0.01) but not c-jun or c-myc mRNA expression. Compared to sham operated controls, [3H]thymidine incorporation was 2.0x, 3.4x, 3.2x, 7.8x, and 2.2x increased following 20%, 35%, 55%, 70%, and 90% PHx, respectively. Serum bilirubin levels remained unchanged until 70% PHx, when they increased from baseline values of 0.54+/-0.05 mg/dl to 1.02+/-0.15 mg/dl (P < 0.05). A further increase occurred following 90% PHx (1.83+/-0.30 mg/dl, P < 0.01). In conclusion these findings indicate that c-fos mRNA expression 15 mins after PHx correlates with hepatic regenerative activity but not the strength of the regenerative stimulus and that hepatic parenchymal loss of 55-70% must occur prior to the detection of elevated serum bilirubin levels. The results also indicate that relative to a 70% PHx, 90% PHx is associated with decreased rather than increased hepatic regenerative activity.

Animals↗

An analysis of early renal transplant protocol biopsies--the high incidence of subclinical tubulitis.

To investigate the possibility that we have been underestimating the true incidence of acute rejection, we began to perform protocol biopsies after kidney transplantation. This analysis looks at the one-week biopsies. Between March 1 and October 1, 1999, 100 adult patients undergoing cadaveric kidney or kidney/pancreas transplantation, or living donor kidney transplantation, underwent 277 biopsies. We focused on the subset of biopsies in patients without delayed graft function (DGF) and with stable or improving renal function, who underwent a biopsy 8.2+/-2.6 d (range 3-18 d) after transplantation (n = 28). Six (21%) patients with no DGF and with stable or improving renal function had borderline histopathology, and 7 (25%) had acute tubulitis on the one-week biopsy. Of the 277 kidney biopsies, there was one (0.4%) serious hemorrhagic complication, in a patient receiving low molecular weight heparin; she ultimately recovered and has normal renal function. Her biopsy showed Banff 1B tubulitis. In patients with stable or improving renal allograft function early after transplantation, subclinical tubulitis may be present in a substantial number of patients. This suggests that the true incidence of rejection may be higher than is clinically appreciated.

Adult↗

Human myeloid dendritic cells transduced with an adenoviral interleukin-10 gene construct inhibit human skin graft rejection in humanized NOD-scid chimeric mice.

Human myeloid DC were generated from peripheral blood mononuclear cells by monocyte adhesion and subsequent culture with rhGM-CSF and rhIL-4. We transduced immature (day 5 of culture) myeloid DC with an E1-deleted replication-deficient adenoviral vector encoding the cytokine IL-10 (AdV IL-10) and a control adenovirus MX-17 (AdV MX 17). Human DC transduced with AdV IL-10 showed inhibition of the mixed leukocyte culture, reduced cell surface expression of co-stimulatory molecules (CD80/CD86) and were unable to produce the potent allo-stimulatory cytokine, interleukin-12. In order to test the in vivo properties of these cells a humanized immunodeficient mouse skin transplantation model was developed. Immunodeficient NOD-scid mice were engrafted with human skin, reconstituted via intraperitoneal injection with allogeneic mononuclear cells (MNC) mixed with 1 x 10(6) DC that were autologous to the skin donor and that had been transduced with either AdV IL-10 or AdV MX-17. Skin grafts were removed at day 7 and 14 after reconstitution and studied histologically for evidence of rejection. In animals that received DC modified with AdV IL-10 there was reduced skin graft rejection as characterized by reduced mononuclear cell infiltration and less dermo-epidermal junction destruction compared with those animals that received DC modified with the control virus alone. Injection of equivalent numbers of donor-derived fibroblasts transduced with AdV IL-10 were ineffective at modifying rejection of skin grafts. Immunosuppressive cytokine gene therapy targeting human DC is a novel means of inhibition of the alloimmune response.

Adenoviridae↗

The community integration measure: development and preliminary validation.

OBJECTIVE: To present a new measure of community integration, the Community Integration Measure (CIM), and to offer preliminary information about its psychometric properties. DESIGN: Validation study. SETTING: Community. PARTICIPANTS: Ninety-two participants placed in 3 subgroups (brain injury survivors, n = 41; significant others, n = 36; college students, n = 15). MAIN OUTCOME MEASURES: The distributional properties, factor structure, internal consistency reliability, content validity, discriminant validity, concurrent validity, and construct validity of the CIM. RESULTS: All items correlated positively with each other and with the total score. Principal components factor analysis confirmed a 1-factor structure, which explained 44.1% of the variance. Internal consistency reliability, using Cronbach's alpha, was.87. Content validity was assured by the development procedure, correspondence with the theoretical model, and direct use of consumer language. Discriminant validity was supported by the CIM's ability to differentiate between subsamples. Criterion validity was supported by using correlations with the Community Integration Questionnaire. Construct validity was supported by correlations with the Interpersonal Support Evaluation List. CONCLUSION: The CIM offers a brief, easily administered measure of community integration that conforms to an empirically derived theoretical model and is psychometrically sound.

Activities of Daily Living↗

Does diabetes target ganglion neurones? Progressive sensory neurone involvement in long-term experimental diabetes.

Targeting of dorsal root ganglia by diabetes could account for the selective sensory abnormalities that patients with early diabetic polyneuropathy develop. In this work, we addressed survival, phenotype and gene expression in sensory neurones in lumbar dorsal root ganglia in a long-term model of experimental streptozotocin-induced diabetes in rats, designed to reflect human disease. Motor and sensory conduction slowing developed early, by the 2-month time point. At 2 months, sensory neurones had no detectable alterations in their calibre or gene expression, assessed using quantitative in situ hybridization studies for mRNA markers that included alpha CGRP, beta CGRP, NFM, t alpha 1-tubulin, SP, VIP, B50 (GAP43), galanin, somatostatin, PACAP, HSP27, c-jun, SNAP 25, p75, TrkA, TrkB and TrkC. By 12 months, however, diabetics had developed neurone perikaryal and distal axon atrophy, accompanied by generalized downregulation of mRNA expression, particularly of CGRP transcripts, PACAP, SP, NFM, p75, trkA and trkC. With the exception of HSP-27, no elevation in mRNAs that increase after injury, such as VIP, galanin, CCK, PACAP, B50 and t alpha 1-tubulin, was observed and constitutive levels, when detectable, trended towards lower rather than increased levels. There was relative preservation of neurone numbers at 12 months; only a non-significant trend towards fewer diabetic neurones was detected using a rigorous and systematic physical dissector counting approach through the entire L5 ganglia. There was no change in the relative populations of CGRP- and SP-immunoreactive neurones. Our findings indicate that even long-term experimental diabetes is associated with relative preservation of sensory neurone populations, but the neurones are atrophic and their gene expression is altered. This pattern of change differs from that following axotomy, implies a degenerative rather than an injury phenotype and has important implications for how such neurones might be rescued.

Animals↗