A radioimmunoassay search for measles and distemper antigens in subacute sclerosing panencephalitis and multiple sclerosis brain tissues

Solid-phase direct sandwich radioimmunoassays have been developed which will detect measles antigen in as little as 20 μg of SSPE brain, measles antigen in 5 × 10⁴ pfu/ml of measles virus, and canine distemper antigen in 5 × 10⁵ pfu/ml of this virus. Using these assays, a search was carried out for measles and distemper antigens in CNS autopsy tissue from 3 multiple sclerosis (MS) patients. Even in 1000-fold higher concentrations of MS brain (20 mg), antigens of neither virus were detected. Additional studies were carried out using similar radioimmunoassays but employing IgG from 2 MS patients as probes against MS brain specimens, including IgG and acute plaque material from the same individual. However, no reaction between MS IgG and MS brain was detected. Thus no evidence was found for foreign antigen in the MS brains nor for autoantibody which reacted with brain tissue antigens.It is concluded that either measles or distemper are not present in acute MS lesions, or that they are present in such small amounts that their detection will require techniques even more sensitive than those used in the present study.     

The use of neurotoxins to characterize the rates and subcellular distributions of axonally transported dopamine-beta-hydroxylase, tyrosine hydroxylase and norepinephrine in the rat brain.

The effect of 6-hydroxydopamine (6-OHDA), colchicine and cytochalasin B on the transport and subcellular distribution of proteins, tyrosine hydroxylase (TH), dopamine-beta-hydroxylase (DBH) and norepinephrine (NE) were studied in the noradrenergic neurons of the rat locus coeruleus (LC). Four waves of transported 3H-labeled proteins and glycoproteins, defined by previous studies, as well as hypothalamic levels of TH, DBH and NE, were examined after injection of each neurotoxin into the ascending dorsal noradrenergic bundle. Blockade of subcellular components of TH, DBH and NE was compared to their endogenous hypothalamic distributions. 6-Hydroxydopamine variably blocked transport of all 4 waves of 3H protein and bilateral injections decreased hypothalamic levels of TH, DBH and NE by 58.2, 56.9 and 52.2% of controls, respectively. Cytochalasin B blocked transport of protein waves I (72--192 mn/day) and III (13--20 mm/day) and decreased hypothalamic levels of TH to 60.1% of control after bilateral injections. Colchicine blocked transport of waves I, II (24--48 mm/day) and V (1.4--2.9 mm/day) and blocked [3H]NE transport, while decreasing hypothalamic levels of DBH and NE to 56.6 and 69.3% of control after bilateral injections. Colchicine and 6-OHDA, but not cytochalasin B, caused a backup of DBH immunofluorescence proximal to the injection site. DBH and NE appeared to be transported primarily in particulate form, while TH transport was predominantly soluble in distribution. None of the toxins differentially affected the transport of one particular subcellular component of TH, DBH or NE. Based on the differential blocking effects of these toxins, DBH and NE appeared to be associated with wave II, and TH with wave III, travelling at 24--48 mm/day and 13--20 mm/day respectively.     

Lymphoid cell infection by measles virus in newborn hamsters : Role for monocytes in virus spread to distant sites

Following intraperitoneal inoculation of measles (HBS) virus into newborn hamsters widespread but variable productive lymphoid tissue infection was detected by a sensitive viral isolation technique. Peritoneal wash cells and spleen were the most common sites of infection. Virus was frequently isolated from the bone marrow and was commonly found in multiple lymph nodes. Thymic infection was only rarely demonstrated. A mononuclear cell associated viremia was demonstrated by Ficoll-hypaque fractionation of peripheral blood from animals receiving both high- and low-dose virus. Infection was present in both plastic adherent and non-adherent fractions of the blood mononuclear cells. The infected cell population in the spleen had both nylon wool and plastic adherent characteristics and infection was therefore thought to be macrophage-associated. The possible relevance of such macrophage associated infection is discussed with regard to the pathogenesis of natural measles infection and the immunosuppression observed in measles-infected hosts.     

Measles antibodies and histocompatibility types in multiple sclerosis

Measles antibody titres and HLA antigens were determined in 71 White and 11 Coloured multiple sclerosis (MS) patients and 71 White and 11 Coloured age and sex-matched controls. Measles antibody titres were determined by the hemagglutination inhibition test and HLA antigens were determined serologically by a micro-lymphocytotoxicity test. Measles antibody titres were significantly higher in MS than in control cases and this was true for both female and male patients. No association was observed between the HLA antigens, especially HLA-A3 or HLA-B7 and measles antibody titres in the sera of MS patients or controls.     

In vitro cellular responsiveness in multiple sclerosis patients to a purified measles virus nuclear core and to other viral antigens

The Clausen modification of the peripheral blood buffy coat leukocyte migration test was used to test patients with multiple sclerosis, normal subjects and patients with other neurologic diseases for cell-mediated immunity to commercial measles virus, purified nuclear core material from a human neurotropic strain of measles virus, as well as to commercial preparations of rubella, mumps and parainfluenza HA₂. Leukocytes of multiple sclerosis patients showed significantly less mean inhibition of migration in the presence of both measles antigens, mumps and parainfluenza but no difference from controls when incubated with rubella. No correlation could be found between the degree of migration inhibition and concomitant serum anti-measles neutralization antibody titers to the same strain of virus. The use of purified viral antigens might result in more meaningful studies of the status of cell-mediated immunity to viral antigens in multiple sclerosis. The relationship of our findings to other studies in this area and to the pathogenesis of multiple sclerosis is discussed.     

Axonal transport of [H]fucosyl glycoproteins in noradrenergic neurons in the rat brain

The transport of [³H]fucosyl glycoproteins has been investigated in the noradrenergic pathway from the locus coeruleus (LC) to the hypothalamus in the rat brain by use of local injection ofl-[6-³H]fucose into the LC. Two discrete waves of³H-glycoprotein pass through the hypothalamus in a caudorostral direction traveling at 4 mm/h (96 mm/day) and 2 mm/h (48 mm/day) respectively. Both waves appear to originate from the LC at approximately 2 h after the injection of [³H]fucose, a time when incorporation into glycoprotein has not yet peaked in the LC. Lesioning the LC-hypothalamic pathway with 6-OHDA, but not carrier solution, blocks both waves of³H-glycoprotein demonstrating that transport occurs exclusively in the catecholamine axons within the pathway. DEAE-Sephadex chromatography combined with 6-OHDA lesions demonstrates clear differences in the character of³H-glycoproteins in the two waves undergoing transport. The relative lack of labelling of hypothalamic glycoproteins in the interval before and after these waves suggests that relatively few rapidly transported glycoproteins contribute to the non-terminal axon membrane and are probably primarily transported to the nerve terminal. No evidence for slow axoplasmic transport of [³H]fucosyl glycoproteins is found.     

Lymphocyte subpopulations in multiple sclerosis: serial studies and clinical correlations.

Total lymphocyte counts and T and SIg+ cell numbers in peripheral blood were determined in 74 healthy controls and 44 MS patients. Twenty-five patients were studied in relapse and at two intervals after ACTH. Nineteen had not relapsed for 6 months but most had progressed clinically. MS patients showed significantly lower total lymphocyte counts which were not correlated with disease activity. Subpopulation analyses showed the MS patients to have significantly lower T cell numbers and significantly elevated SIg+ cell numbers and percentages. Although T cell numbers were low in all phases, both T and SIg+ cell abnormalities were maximal in progressive disease or in the recovery phase after relapse and a significant fall in T cell percentage and rise in SIg+ cell number was confirmed in serial studies of 16 patients followed from relapse to recovery. SIg+ cell numbers correlated negatively with relapse rate and correlated positively with the ratio of disability to number of relapses. The incidence in MS patients of HLA A3 was significantly increased and of HLA B12 and HLA BW40 significantly decreased but no correlation was seen between HLA phenotype and T, SIg+ or total lymphocyte counts. It is suggested that the peripheral blood SIg+ cell abnormalities and some of the T cell abnormalities are secondary to the pathological process, perhaps due to antigenic stimulation and particularly in non-relapsing progressive disease. Their possible role in influencing recovery or progression is discussed.     

Geniculate interactions with a penicillin discharge in visual cortex

Spontaneous activity of principal cells in the dorsal nucleus of the lateral geniculate body was studied 1 sec before and 1 sec after penicillin discharges in the striate cortex. Many cells exhibited increased activity prior to the penicillin discharge suggesting that corticopetal activity from the geniculate is capable of triggering the penicillin discharge. Many cells also exhibited increased activity for as long as 1 sec after the discharge. Some cells displayed both phenomena. Two patterns of transient depression were also observed in the postdischarge period. The latter phenomena are believed to be related to the inhibitory process associated with after-positivity which follows orthodromic or antidromic stimulation of the geniculate body.     

Some actions of catechol on synaptic transmission in the isolated spinal cord of the frog.

The isolated frog spinal cord was used to investigate the synaptic effects of the convulsant agent catechol. Addition of the compound to the superfusate consistently enhanced orthodromic reflex activity recorded from ventral roots and augmented primary afferent depolarization. Concomitantly catechol altered the polarization changes produced in ventral and dorsal roots by putative neurotransmitter amino acids when these compounds were applied in Mg2+-containing Ringer. Catechol reduced the hyperpolarizations induced in motoneurons by the neutral amino acids, GABA, beta-alanine, taurine and glycine, but did not affect the depolarizations produced by the dicarboxylic amino acids, L-glutamate and L-aspartate. In contrast, catechol increased the dorsal root depolarizations elicited by both neutral and dicarboxylic amino acids and also the depolarizations produced by elevated potassium concentrations. Catechol did not bring about significant changes in the passive electrical properties of motoneurons or dorsal root fibers. In addition, it did not alter either the high affinity uptake or the depolarization-evoked release of tritiated GABA, glycine, L-glutamate and L-aspartate. It appears that the postsynaptic actions of catechol explain its ability to enhance spinal reflexes.     

Endogenous myelin basic protein-serum factors (MBP-SFS) and anti-MBP antibodies in a patient with post-herpes simplex virus acute disseminated encephalomyelitis

The measurement of myelin basic protein serum factors (MBP-SFs) and anti-MBP antibodies in specimens from a patient with post-herpes simplex acute disseminated encephalomyelitis (ADE) is described.

Transitory appearance of high affinity anti-MBP antibodies in the absence of detectable MBP-SFs was observed. This pattern was similar to that found previously in acute multiple sclerosis and experimental allergic encephalomyelitis.

These findings are consistent with the hypothesis that a possible normal role of MBP-SFs is as neuroautotolerogens, preventing autoreactive lymphocyte clones from damaging myelin.     

Electron-microscopic observations on preterminal fibers in the oculomotor nucleus of the cat. With special reference to the relation between axon diameter and myelin thickness in mammalian gray matter.

Electron microscopy of myelinated fibers in main lateral cell groups of the cat oculomotor nucleus reveals the majority to have diameters of less than 3 mum, with more than one-half less than 1 mum in diameter. The values of the ratio g (axon diameter/total fiber diameter) range between 0.58 and 0.88, with a mean of 0.74. There is no definite relation between g and fiber diameter. The morphological data are consistent with physiological data on preterminal conduction velocities in the oculomotor nucleus, and extend the dimensional analysis of myelinated fibers in gray matter, which has previously been confined to lower vertebrates, to the mammalian central nervous system.     

Heroin vs alcohol addiction--quantifiable psychosocial similarities and differences

A pilot study of 50 heroin addicts and 66 alcohol addicts utilizing the recently developed quantitative methods for measuring life change and seriousness of illness. Alcohol addicts were found to have significant attenuation of perception of life change and seriousness of illness as compared to heroin addicts and current “normative” data. The heroin addicts were found to have a mild but significant augmentation of perception of life change and seriousness of illness as compared to the “normative” data. In addition both the heroin addicts and alcohol addicts were found to maintain very high levels of life change. The documented differences in perception and the maintenance of high levels of life change were felt to have potential significance in the treatment and evaluation of both groups of addicts.     

Probability of conduction deficit as related to fiber length in random-distribution models of peripheral neuropathies.

This paper presents a set of probabilistic models which reproduce the proximodistal gradient of sensory deficit in peripheral neuropathies, on the basis of the occurrence of axonal dysfunction as a result of randomly distributed abnormalities. The models, which are based on conduction block, loss of temporal coherence, and weak interactions between nerve fibers, demonstrate that randomly distributed axonal dysfunction provides a sufficient condition for distal sensory deficit. The models predict a marked reduction in the length for normal sensory conduction with small increases in the probability of axomal dysfunction, providing a possible correlate for the rapid clinical progression of some neuropathies. The hypothesis that weak interactions between fibers result in paresthesiae in peripheral neuropathies is also discussed.     

Scaling of life events by psychiatric patients and normals.

Life event scaling was investigated in 171 male psychiatric patients, a comparison group of 181 “normals”, and 165 relatives of patients and normals. The social readjustment rating questionnaire (SRRQ) of Holmes and Rahe was employed as the scaling instrument, and a social readjustment rating scale (SRRS) was calculated for all groups. Although psychiatric patients and normals agreed on the rank ordering of events, patients generally assigned greater magnitudes to items than did normals. The most significant scaling differences were for items of a martial-family or personal nature. Future research concerning the relationship of life events and psychiatric symptoms should consider employing SRRS weights derived from psychiatric populations rather than existing scores from normative groups.     

Species cross-reactivities of antisera reacting with myelin basic peptide 43–88 : Applications for the detection of peptide in rabbit plasma

Previous investigations have demonstrated that the detection and measurement of peptides of myelin basic protein (BP) by immunochemical procedures are heavily dependent on the conformation of BP peptides and the specificities of antisera to these peptides. In the present study a double antibody-radioimmunoassay was used to assess the species specificity of antisera from 4 rabbits and 2 sheep against myelin basic protein (BP) peptide 43-88. Although some antisera showed broad reactivity, others were selective for species of BP used in the immunogen. By optimizing the reaction of one of the sheep anti-human peptide 43-88 for detection of rabbit BP peptide 43-88, it was possible to detect immunoreactive BP peptide 43-88 in the plasma of rabbits previously injected with bovine BP peptide 43-88 and complete Freund's adjuvant (CFA). Peptide could be detected in plasma for 1-2 days immediately after subcutaneous injection and again between days 20 and 50. Plasma antibody to rabbit BP peptide 43-88 also appeared 15-20 days after immunization and persisted often in an oscillatory pattern reciprocal for the peptide level. Neither peptide nor antibody was detected in the plasma of normal rabbits or of rabbits injected with bovine spinal cord homogenate-CFA, bovine renal homogenate-CFA or CFA alone. There was no relationship of the level of immunoreactive BP peptide 43-88, antibody to this peptide or their patterns to clinical or neuropathological changes in the animals. Quantitation and analysis of BP peptides in body fluids of animals with diseases affecting central nervous system myelin may provide information relevant to human demyelinating diseases.     

Language in the absence of inner speech

A 54-yr-old man abruptly became mute with mild right hemiparesis. Written expression, comprehension of speech and print, calculating ability, and verbal short-term recall, although slow, were largely preserved. Extensive studies demonstrated complete loss of inner speech. He was unable to speak to himself and unable to appreciate the phonological structure of words. Instead, his preserved language skills were based on a highly developed visual imagery. This case greatly extends the range of language skills previously thought possible in individuals who suffer the loss of inner speech.     

Virus antibodies in Parkinson's disease: Herpes simplex and measles virus antibodies in serum and CSF and their relation to HLA types

Serum and cerebrospinal fluid (CSF) immunoglobulin G (IgG) antibodies to herpes simplex (HSV) and measles viruses were assayed with a radioimmunoassay in 56 patients with idiopathic Parkinson's disease and in a similar number of age- and sex-matched controls with other neurological diseases. As a group, the patients with Parkinson's disease had a significantly increased serum antibody level against HSV, but measles virus antibody levels were similar in both groups. Both in the Parkinson's group and in the control group, the levels of the total IgG in CSF were within normal limits and the CSF antibodies to HSV and measles virus paralleled the serum antibody titers relative to the total IgG serum-to-CSF ratios. This indicates no increased intrathecal antibody production in either group. In 48 patients with Parkinson's disease who were HLA-typed, no association of viral antibody levels with particular HLA antigens were noted.The findings suggest that HSV is not present within the central nervous system of the patients with Parkinson's disease. The increased HSV antibody level seen in Parkinson's disease patients may reflect a more general disturbance of the patients' immune functions.     

Morphology and distribution of cytoplasmic inclusions in adrenoleukodystrophy

We report some additional observations on the pathognomonic cytoplasmic inclusions in a typical case of adrenoleukodystrophy (ALD). Study of the linear inclusions by means of tilt-stage electron microscopy indicated that they represent trilaminar sheet-like structures distinct from materials seen in various other storage disorders. This study emphasizes that the inclusions in the adrenal and testicular parenchymal cells morphologically differ from those in the brain macrophages. In the latter they occur as membrane-bound discrete structures while in the parenchymal cells they are always seen lying free in the cytoplasm and associated with cleft-like spaces. This morphologic variation of the inclusions is noteworthy and probably reflects a biochemical difference in the storage materials in these two most severely affected organs. In addition, this study demonstrates the presence of linear inclusions within macrophages in otherwise normal-appearing liver, spleen, thymus and also in the renal tubular cells. These findings suggest that the abnormal materials containing very long chain fatty acids enter the blood stream and become sequestered in the reticuloendothelial system and possibly are also excreted in the urine. Detection of these abnormal fatty acids in blood and urine may simplify diagnosis of ALD.     

Axoplasmic transport of norepinephrine in the locus coeruleus-hypothalamic system in the rat.

The axoplasmic transport of norepinephrine (NE) between the nucleus locus coeruleus (LC) and the hypothalamus has been measured by three methods which employ the stereotaxic injection of [3H]catecholamines in the rat brain. [3H]NE is synthesized from [3H]dopamine (DA) injected into the LC and transported at a rate of 0.8-0.9 mm/h. This rate is probably an underestimate since it includes the time required for uptake and conversion of [3H]DA to [3H]NE prior to transport. Measurement of transport of [3H]NE between the posterior and anterior hypothalamus gives a rate of 2 mm/h which is independent of [3H]DA uptake and [3H]NE synthesis. This rate is in good agreement with the 1.9 mm/h figure calculated for [3H]NE transport in the system after LC injection of [3H]NE and represents a closer estimate of the true rate of axoplasmic transport than the 0.8-0.9 mm/h estimate. Transport of [3H]NE occurs through primarily ipsilateral NE fibers running in the median forebrain bundle and is blocked by 6-hydroxydopamine lesions in the bundle. Early appearing [3H]NE in the hypothalamus forms a relatively stable pool which is unaffected by median forebrain bundle lesions and is essentially unchanged by [3H]NE transported from the LC. This pool appears to be derived from non-specific spread of [3H]DA after injection, probably through the cerebrospinal fluid.