The specificity of 26 monoclonal antibodies against the CA 125 antigen was investigated in two phases of the ISOBM TD-1 workshop. The binding specificity was studied using CA 125 immunoextracted by specific antibodies immobilized on various solid phases, or on the surface of human cell lines. Immunometric assays using all possible antibody combinations were used to study the topography of antibody binding sites on the antigen. We conclude that the CA 125 antigen carries only two major anti-genic domains, which classifies the antibodies as OC125-like (group A) or Mil-like (group B). One antibody, OV 197, showed binding specificity related to some of the OC125-like antibododies, but was classified into a separate group C. The OC125-like group of antibodies has four subgroups with different binding specificities. These are A1 = OC 125 and K 95, A2 = K 93, A3 = B43.13, and A4 = ZS 33, B27.1 and CCD 247. Binding of nonlabelled OC 125 or K 95 to CA 125 caused a marked increase in binding of labelled OV 197 to the complex. This conformational change was not observed with any other antibody combinations. Antibody B43.13 could form immunometric assay combinations particularly with antibodies of subgroup A4, indicating that the B43.13 epitope is in the periphery of the binding area of OC125-like antibodies. The M11 -like group of antibodies is more homogenous with strong cross-inhibition between most antibodies. Only one antibody, ZR 38, would form an immunoassay combination with other M11 -like antibodies and thus represents a distinct subgroup. The main group of M11-like antibodies are M 11, ZR 45, MA602-6, K 91, OV 185, K 101, K 90, K 96, K 97, K 102, CCD 242, 145-9, and 130-22. Antibody OV 197 binds to a domain designated C and is unique, as stated above. Antibody pairs from any two of the three groups may be used in immunometric assays. Three antibodies were not studied by complete cross-inhibition due to low affinity (OV 198 and K 100) or lack of material (MA602-1). OV 198 and K 100 are most likely OC125-like and MA602-1 is Ml 1-like. Antibody affinity was estimated with labelled antigen in solution or with antigen adsorbed on microtiter wells. Western blot analysis showed staining both in the stacking gel and corresponding to a molecule of 200 kDa. There was a marked difference between the antibodies in their ability to bind to CA 125 immobilized on a membrane. Strongest binding was observed with the M11-like antibodies, particularly M 11, K 96, K 97, MA602-6, 145-9. Antibodies belonging to the subgroup A4 were the only OC 125-like antibodies which reacted well with CA 125 in Western analysis. Digestion of CA 125 with proteolytic enzymes showed it to be particularly sensitive to trypsin cleavage. However, no low molecular weight fragments with preserved immunoreactivity were found.

1. Single-unit recordings were obtained from the supplementary motor area of the cerebral cortex of two monkeys during execution of learned movements 2. Monkeys were required to push or pull a cast attached to the right forelimb in response to a sudden perturbation delivered via the cast. An instruction as to the direction of the monkey's movement was delivered 2.5--5 s prior to the occurrence of the perturbation and correct performance, therefore, required the animal to develop a preparatory state prior to the perturbation. 3. Of many hundreds of neurons recorded, 201 exhibited instruction-induced changes of activity during the period intervening between the instruction and the perturbation-triggered movement. 4. In 94 neurons, effects of the instruction were differential depending on which of the two instructions was given, whereas in 107 neurons, effects were nondifferential. The latencies of the differential responses appeared to be shorter (starting as early as 140 ms after the instruction). 5. The magnitude of the instruction effects varied in parallel with development of enhanced motor skill as the monkeys gained more experience in responding to the triggering stimulus. 6. These observations substantiate the hypothesis that the supplementary motor area plays a part in modifying a sensory-triggered motor output.

The study and clinical assessment of brain disease is currently hindered by a lack of non-invasive methods for the detailed and accurate evaluation of cerebral vascular pathology. Angiography can detect aberrant flow in larger feeding arteries/arterioles but cannot resolve the micro-vascular network. Small vessels are a key site of vascular pathology that can lead to haemorrhage and infarction, which may in turn trigger or exacerbate neurodegenerative processes. In this study, we describe a method to investigate microvascular flow anisotropy using a hybrid arterial spin labelling and multi-direction diffusion-weighted MRI sequence. We present evidence that the technique is sensitive to the mean/predominant direction of microvascular flow in localised regions of the rat cortex. The data provide proof of principle for a novel and non-invasive imaging tool to investigate cerebral micro-vascular flow patterns in healthy and disease states.

Glial scar formation is the major impedance to axonal regrowth after spinal cord injury (SCI), and scar-modulating treatments have become a leading therapeutic goal for SCI treatment. In this study, human neural stem cells (NSCs) encoding interferon-β (INF-β) gene were administered intravenously to mice 1 week after SCI. Animals receiving NSCs encoding IFN-β exhibited significant neurobehavioral improvement, electrophysiological recovery, suppressed glial scar formation, and preservation of nerve fibers in lesioned spinal cord. Systemic evaluation of SCI gliosis lesion site with lesion-specific microdissection, genome-wide microarray, and MetaCore pathway analysis identified upregulation of toll-like receptor 4 (TLR4) in SCI gliosis lesion site, and this led us to focus on TLR4 signaling in reactive astrocytes. Examination of primary astrocytes from TLR4 knockout mice, and in vivo inhibition of TLR4, revealed that the effect of IFN-β on the suppression of glial scar formation in SCI requires TLR4 stimulation. These results suggest that IFN-β delivery via intravenous injection of NSCs following SCI inhibits glial scar formation in spinal cord through stimulation of TLR4 signaling.

Functional neuroimaging, applied to pre-clinical models of chronic pain, offers unique advantages in the drive to discover new treatments for this prevalent and oppressive condition. The high spatial and temporal resolution of fMRI affords detailed mapping of regional pharmacodynamics that underlie mechanisms of pain suppression by new analgesics. Despite evidence supporting the translational relevance of this approach, relatively few studies have investigated fMRI abnormalities in rodent models of chronic pain. In this study, we used fMRI to map the BOLD response in a recently developed putative rat model of fibromyalgia to innocuous and acute nociceptive stimuli by applying a step-wise graded electrical forepaw stimulation paradigm, with comparison to healthy controls. We observed discriminatory functional signatures (p < 0.001) to 2 mA electrical forepaw stimulation, found to be innocuous in the control group. As such, this translational approach provides sensitive and quantitative neural correlates of the underlying chronic disease. The regional patterns of functional augmentation were found to be concordant with previous studies of nociception in the anaesthetised rat brain, supporting the specificity of this approach in the study of altered central pain processing in reserpine induced myalgia. The methodology introduced in this work represents a novel platform for emerging treatment evaluation in highly experimentally controlled conditions.