Licence for Publication:The Corresponding Author has the right to grant on behalf of all authors and does grant on behalf of all authors, an exclusive licence (or non exclusive for government employees) on a worldwide basis to the BMJ Publishing Group Ltd to permit this article (if accepted) to be published in PMJ and any other BMJPGL products and sublicences such use and exploit all subsidiary rights, as set out in our licence. 2 AbstractBackground: In the chronic stage of stroke, previous work has shown that the worse

The expression of the neurofilament proteins of the goldfish visual pathway reflects the degeneration and regeneration of the optic nerve after nerve crush. To monitor these processes, monoclonal antibodies (mAb) were generated to the intermediate filament proteins of this pathway. The predominant goldfish visual pathway intermediate filament proteins have a molecular weight of 58K and can be separated into 4 isoelectric variants, 2 of which are neuronal (ON1 and ON2) and 2 of which are non-neuronal (ON3 and ON4). The specificities of the mAbs were characterized biochemically and histologically. Immunoblot analysis demonstrated that 2 of the antibodies reacted specifically with the neurofilament proteins (ON1/ON2) and another antibody reacted specifically with the glial filament proteins (ON3/ON4) and with a 48K optic nerve protein of non-neuronal origin. Chymotrypsin digestion of the ON proteins and immunoblotting of the resulting fragments showed that the anti-ON1/ON2 mAbs were directed toward the variable domains of the filament proteins. In contrast, the anti-ON3/ON4 mAb was directed toward the 40K chymotrypsin-resistant region of the glial filament proteins containing the conserved intermediate filament core. When sections of optic nerve tissue were incubated with anti-ON1/ON2 or anti-ON3/ON4 mAbs, the staining resulted in either axonal or glial patterns, respectively. In retina, after optic nerve crush, anti-ON1/ON2 labeled retinal ganglion cells and Müller fibers. In contrast, prior to optic nerve crush, only Müller fibers were labeled. One of the neuronal-directed mAbs was used to decorate growing neurites from retinal explants; anti-ON1/ON2 reactivity appeared in a time-dependent manner that paralleled the expression of ON1/ON2 in vivo. Thus, the antibodies can differentiate these 2 types of goldfish intermediate filament proteins and can be used to monitor optic nerve regeneration in the goldfish visual pathway both in vivo and in vitro.

This study assessed the impact of environmental cadmium and lead exposure on the immune system of more than 2000 children and adults. Serum immunoglobulins [immunoglobulins (Ig) A, G, and M] and peripheral blood lymphocyte phenotypes (T cells, B cells, NK cells, and CD4/CD8 subsets) were measured in a total of 2041 children and adults who lived either in sites with elevated soil levels of cadmium and lead (n = 1561) or in comparison communities (n = 480). The blood lead and urine cadmium levels of participants were somewhat higher than national averages. Mean blood lead levels were 7 microg/dl for participants aged 6-35 mo; 6 microg/dl for participants aged 36-71 mo, 4 microg/dl for participants aged 6-15 yr; and 4.3 microg/dl for participants aged 16-75 yr. Multivariate analysis indicated no marked differences in any of the immune marker distributions attributed to lead for adults or children over 3 yr of age. However, in children under age 3, increased blood lead levels, principally those over 15 microg/dl, were associated with increases in IgA, IgG, IgM, and circulating B lymphocytes. Among adults, urine cadmium levels over 1.5 microg/g were associated with higher levels of IgA and circulating B lymphocytes. No evidence of immunosuppression was noted. The findings of potential immunologic effects at lead levels > 15 microg/dl in young children and at urine cadmium levels > 1.5 microg/g in adults are interesting, but too few participants had these high levels to delineate a threshold. Therefore, we find these results intriguing, but requiring confirmation in populations with higher exposure levels.

Background and ObjectivesThe loss of noradrenergic neurons in the locus coeruleus (LC) contributes to various cognitive and neuropsychiatric symptoms in Parkinson’s disease (PD) and progressive supranuclear palsy (PSP). The spatial precision of in vivo LC imaging is improved using a magnetisation transfer sequence combined with ultra-high field 7T MRI. This study aimed to test the sensitivity of LC imaging in PD and PSP, to characterise the spatial pattern of LC atrophy in patients, and its relationship to cognition and apathy.MethodsThis cross-sectional observational study recruited patients with idiopathic PD, probable PSP-Richardson’s syndrome and age-matched healthy controls (HC) via specialist clinics and volunteer registries. All participants underwent clinical assessments for cognition and apathy, and high-resolution (0.08 mm3) LC scans. To quantify LC integrity, the contrast-to-noise ratio (CNR) relative to a pons reference region was calculated and extracted using a probabilistic atlas. Subregional mean CNRs were summarised to test group differences and to correlate LC integrity with apathy and cognition scores. LC clusters were identified to confirm the spatial pattern of the effect (threshold free cluster enhancement, 10000 permutations, p<0.05, corrected for family-wise error).ResultsTwenty-five patients with PD, 14 with PSP and 24 controls with completed dataset were included in the study. Patients with PSP were more impaired on global cognition and apathy scores, compared to controls and PD. Clusters with reduced contrast were observed in the caudal LC for both PD and PSP patients relative to controls (HC>PD, right caudal LC, 37 voxels; HC>PSP, bilateral caudal LC, 206 voxels). PSP and PD patients showed similar levels of LC degeneration, but this was more extensive in PSP. Across both disease groups, LC integrity was associated with cognitive performance (MoCA: F(1,37)=5.339, p=0.027, bilateral LC, 200 voxels; ACE-R: F(1,37)=4.297, p=0.045, left LC, 70 voxels) and apathy (Apathy Scale: F(1,37)=4.335, p=0.044, left LC, 99 voxels).DiscussionWe confirm the sensitivity of 7T LC imaging to detect sub-regional LC changes in PD and PSP. The relationship between LC integrity and non-motor symptoms highlights the potential for noradrenergic therapeutic strategies to ameliorate cognitive and behavioural features of PD and PSP.