MS Abstracts 8c-2g1

Multiple Sclerosis (MS) is an Inflammatory DeMyelinating Disease of the Central Nervous System that causes Motor, Sensory, and Cognitive Deficits.

The present study characterized DeMyelinated lesions in the Cerebral Cortex of MS patients.

One hundred twelve Cortical lesions were identified in 110 tissue blocks from 50 MS patients. Three Cortical DeMyelination patterns identified:

1. Type I lesions were contiguous with
   • SubCortical White Matter lesions
2. Type II lesions were small
   • Confined to the Cortex, often PeriVascular
3. Type III lesions extended from
   • The Pial surface to Cortical layer 3 or 4

Inflammation and Neuronal pathology were studied in tissue from 8 and 7 patients, respectively.

Compared to White Matter lesions, Cortical lesions contained 13 times fewer CD3-positive Lymphocytes (195 vs 2,596/mm3 of tissue) and 6 times fewer CD68-positive Microglia/Macrophages (11,948 vs 67,956/mm3 of tissue).

Transected Neurites (both Axons and Dendrites) occurred at a density of:

• 4,119/mm3 in active Cortical lesions
• 1,107/mm3 in chronic active Cortical lesions
• 25/mm3 in chronic inactive Cortical lesions
• 8/mm3 in Myelinated MS Cortex
• 1/mm3 in control Cortex

In active and chronic active Cortical lesions, activated Microglia closely apposed and ensheathed Apical Dendrites, Neurites, and Neuronal Perikarya.

In addition, Apoptotic Neurons were increased significantly in DeMyelinated Cortex compared to Myelinated Cortex.

These data support the hypothesis that DeMyelination, Axonal transection, Dendritic transection, and Apoptotic loss of Neurons in the Cerebral Cortex contribute to Neurological dysfunction in MS patients.

The proliferation marker Ki-67 labels cell nuclei in the G(1), S, M, and G(2) phases of the cell cycle.

We used Ki-67 ImmunoHistoChemistry to quantify proliferating Glial Cells in Brain tissue sections from twenty-four patients, comprised of Multiple Sclerosis, normal Brains, and Other Neurological Disease controls.

Glial proliferation was greatly increased in MS Lesions when compared with control Brain White Matter.

Both actively DeMyelinating/early ReMyelinating plaques and chronic inactive plaques of long standing often displayed large numbers of Glial Cells in the proliferative cycle.

The bulk of these proliferating cells were of Oligodendroglial lineage in the MS plaques. Ki-67 positive Macrophage/Microglial lineage cells were largely restricted to acute lesions.

The finding of increased numbers of proliferating Oligodendroglia in most MS plaques, regardless of disease duration or activity state, indicates that the MS Brain is capable of recruiting unexpectedly large numbers of new Oligodendrocytes over long periods of time.

The factors within the MS plaque MicroEnvironment that provoke new Oligodendrocyte generation and their subsequent loss still need to be identified.

Copyright 2001 Wiley-Liss, Inc.

The purpose of the study was to investigate differences in contractile speed, force, and Fatigability of the Adductor Pollicis Muscle between 12 patients with Multiple Sclerosis (MS) and 8 sedentary control subjects matched for age and gender.

There were no differences between the patients with MS and control subjects with respect to the percentage of maximal muscle force that could be recruited during voluntary effort (95.5 +/- 3.9% and 98.2 +/- 2.0%, respectively, P = 0.10).

The stimulation frequency/force and force/velocity relationships, the rates of force development and relaxation, fatigue resistance, and the recovery rate of Adductor Pollicis Muscle.

However, previous results from the same group of MS patients showed that Quadriceps Femoris Muscle force and resistance to fatigue were reduced.

Therefore, our data support the clinical experience that, in patients with MS, lower limb muscle function is more or earlier affected than upper limb muscle function.

Copyright 2001 John Wiley & Sons, Inc.

Objective
ß-Chemokines were recently demonstrated in active MS-lesions.

We tested whether MCP-1 and RANTES can also be detected in CSF and Serum of patients with MS and whether release is associated with Inflammatory Disease activity.

Materials And Methods
CSF and Serum from 34 patients with newly diagnosed Relapsing/Remitting MS (RR-MS), 17 patients with Viral Meningitis (VM) and 19 patients with Non-Inflammatory Neurological Diseases (NIND) were investigated by ELISA.

RR-MS patients underwent Lumbar Puncture and Gd-enhanced MRI examinations within 2 days.

Results
MCP-1 was strong Intrathecally released in all patients.

Compared to NIND CSF-levels were increased in VM (P<0.001) and were decreased in RR-MS (P<0.05). RANTES was only detected in Serum in all patients.

Levels were higher in VM and RR-MS compared to NIND (P<0.05). A total of 14/34 RR-MS patients exhibited active Gd-enhancing Lesions on MRI.

They had lower MCP-1 levels in CSF (P<0.001) and Serum (P<0.05) and higher Serum levels of RANTES (P<0.05) as compared to patients without active lesions.

Conclusions
MCP-1 and RANTES are differentially released during acute attacks of RR-MS, which might reflect different ImmunRegulatory roles of these ß-Chemokines in RR-MS.

Objective
To determine the frequency and significance of AntiNuclear (ANA), AntiCardiolipin (ACA) and Anti-Ro (SS-A) AntiBodies in Multiple Sclerosis (MS) patients.

Methods
ANA (indirect ImmunoFluorescence), ACA and anti-Ro (SS-A) AntiBodies (ELISA) were tested in Sera of 42 patients with Poser defined MS and 50 healthy individuals.

Results
High levels of anti-Ro (SS-A) AntiBodies were found in 3 patients (7%) (vs 0 in the control group).

Two of them had normal salivary gland biopsy. Clinical MS form was Chronic/Progressive in 2 cases and Relapsing/Remitting in the third one.

Ten patients (23%) had low levels of ANA (vs 4%), none of them positive for Anti-Ro (SS-A) AntiBodies.

Only 1 patient (2%) with RR clinical form had ACA (vs 0). No clinical or NeuroRadiological differences with conventional MS patients were observed.

Conclusions
ANA, ACA and Anti-Ro (SS-A) AntiBodies in MS patients indicate an underlying AutoImmune Disease but our series suggests that they are an Epiphenomenon of a more diffuse Immunological dysfunction.

Proton Magnetic-Resonance Spectroscopy (PMRS) was used to measure the levels of Inositol/MyoInositol (Ins), Choline, Creatine/Phosphocreatine (Cr), Glutamine/Glutamate (Glx/Glx1), N-AcetylAspartate (NAA), Gamma-AminoButyric Acid (GABA).

And Lipids were measured in the foci of DeMyelination in the Brains of 59 patients with Multiple Sclerosis (MS). Magnetic-Resonance Imaging was performed during a single investigation.

A control group comprised 20 healthy individuals. PMRS revealed significant alterations in the levels of metabolite in all the patients as compared with the controls.

Decreases in NAA by 23-52%, in Cr by 12-21%, in Choline by 15-26%; increases in Ins by 51-63%; as well as the appearance of Lipids (up to 100%)

In MS, there were reduction in NAA/Cr, NAA/Choline, and NAA/Choline/Cr ratios by 12-53; 10-19; and 57-82%, respectively.

As compared with the Remitting MS, Secondary/Progressive MS showed decreases in the content of NAA by 23-25%, NAA/(Choline + Cr) by 48-54% and increases in the levels of Ins and Lipids by 50-76%.

In Remitting MS, there was a strong correlation between the NAA/Cr ratio and the volume Brain lesion.

It is concluded that PMRS evaluated the extent, pattern and activity of DeMyelination (by the levels of Ins, NAA, Cr, Lipids) and the intensity of Cerebral Atrophy (by NAA levels, NAA/Cr ratio).

The findings testify that there are NeuroChemical differences between Remitting and Secondary/Progressive MS.

Background And Purpose
Current imaging does not permit quantification of Neural injury after Traumatic Brain Injury (TBI) and therefore limits both the development of new treatments and the appropriate counseling of patients concerning prognosis.

We evaluated the utility of Magnetization Transfer Ratio (MTR) and Proton MR Spectroscopy in identifying patients with Neuronal injury after TBI.

Methods
Thirty patients with TBI (21-77 years old; mean age, 42 years; admission Glasgow Coma Scale (GOS) scores 3-15; mean score, 11) were studied on a 1.5-T system with Magnetization Transfer imaging and MR Spectroscopy of the Splenium.

Magnetization Transfer imaging was also performed in the BrainStem in all patients, and other areas of the Brain were sampled in one patient.

The Splenium of the Corpus Callosum and BrainStem were studied because these are often affected by diffuse Axonal injury.

Scans were obtained 2 to 1129 days after injury (median, 41 days). MTR was considered abnormal if it was more than 2 SD below normal.

Proton MR Spectroscopy was used to calculate the N-AcetylAspartate (NAA)/Creatine (Cr) ratio. GOS was determined at least 3 months after injury.

Results
In 10 patients with a GOS of 1 to 4, the mean NAA/Cr was 1.24 +/- 0.28; two of these patients had abnormal MTR in Normal-Appearing White Matter (NAWM).

In 20 patients with a GOS of 5, the mean NAA/Cr was 1.53 +/- 0.37 (P < .05); four of these patients had abnormal MTR in NAWM.

MTR abnormalities in NAWM were identified in six patients, but these changes did not correlate with GOS or MR Spectroscopy changes.

Conclusion
MTR and MR Spectroscopy can quantify damage after TBI, and NAA levels may be a sensitive indicator of the Neuronal damage that results in a worse clinical outcome.

The aim of this study was to conduct an Ecological analysis of the extent to which UltraViolet Radiation (UVR) levels might explain the regional variation of Multiple Sclerosis (MS) in Australia.

MS prevalence data for six Australian regions were compared with UVR levels of the largest city in each region, with some other climatic variables and with the Melanoma incidence in the same regions.

A close association was found between the theoretical MS prevalence predicted from UVR levels and the actual prevalence.

Furthermore, the negative correlation between UVR and MS prevalence (r = -0.91, p = 0.01) was higher than the positive correlation observed for UVR and malignant Melanoma incidence (r = 0.75, p = 0.15 for males and r = 0.80, p = 0.10 for females).

This study demonstrated that the regional variation in MS prevalence in the continent of Australia could be closely predicted by regional UVR levels.

It is consistent with the hypothesis that UVR exposure may reduce the risk of MS possibly via T-Lymphocyte-mediated ImmunoSuppression.

Analytical Epidemiology studies are required to investigate this specific hypothesis.

Copyright 2001 S. Karger AG, Basel

Trigeminal Neuralgia is a well-recognized complication of Multiple Sclerosis. In patients with Neuralgia not responding to medical treatment or transcutaneous ablative procedures, the pain can often be treated successfully by partial Rhizotomy of the Trigeminal Sensory Root.

We have examined partial Trigeminal Rhizotomy specimens from six Multiple Sclerosis patients, aged between 34 and 77 years, with intractable Trigeminal Neuralgia lasting between 18 months and 11 years.

The Rhizotomy specimens were placed in buffered Glutaraldehyde immediately after resection, and subsequently processed for electron microscopy.

In all cases, this revealed DeMyelination in the proximal (CNS) part of the Nerve Root, with associated Gliosis and variable inflammation.

A consistent feature was the presence of clusters of juxtaposed Axons without intervening Glial processes.

Similar juxtaposition of Axons was previously observed in Trigeminal Neuralgia due to Vascular compression of the Nerve Root.

Experimental studies indicate that this arrangement of DeMyelinated Axons is conducive to both spontaneous Impulse activity and Ephaptic spread of excitation.

The DeMyelination and associated juxtaposition of Axons may therefore account for key aspects of the PathoGenesis of Trigeminal Neuralgia.

Theiler's Virus is a PicornaVirus responsible for a persistent infection of the Central Nervous System of the mouse, leading to a chronic DeMyelinating Disease considered to be a model for Multiple Sclerosis.

The leader (L) protein encoded by Theiler's Virus is a 76-Amino-Acid-long Peptide containing a Zinc-binding motif.

This motif is conserved in the L proteins of all CardioViruses, including EncephaloMyoCarditis Virus.

The L protein of Theiler's Virus was suggested to interfere with the /ß-Interferon (IFN-alpha/ß) response (W.-P. Kong, G. D. Ghadge, and R. P. Roos, Proc. Natl. Acad. Sci. USA 91:1796-1800, 1994).

We show that expression of the L protein indeed inhibits the production of /ß--Interferon by infected L929 cells.

The L protein specifically inhibits the transcription of the IFN-4 and IFN-ß Genes, which are known to be activated early in response to Viral infection.

Mutation of the Zinc finger was sufficient to block the Anti-Interferon activity, outlining the importance of this motif in the L protein function.

In agreement with the Anti-Interferon role of the L protein, a Virus bearing a mutation in the Zinc-binding motif was dramatically impaired in its ability to persist in the Central Nervous System of SJL/J mice.

The relative distribution of Gangliosides was determined in the Serum of 37 patients with Multiple Sclerosis (MS) and of 30 healthy subjects.

There was a significant increase of GM1 and GD1a, and a decrease of GM3 proportion in the Serum of Relapsing/Remitting MS patients (RRMS) during their first MS attack.

The RRMS patients in relapse with a long duration of the disease had a significant decrease of GM1 and an increase of GD1a portion in the Serum.

An increase of GD1a, one of the major Brain Neuron Ganglioside fraction, suggested the Neuron injury in the early and with a long duration RRMS.

The finding of an increase of GM1, the main human Myelin Ganglioside, during the first MS attack in RRMS patients confirms previous evidence for the possible involvement of Gangliosides in the early pathological course of DeMyelination in MS.

ß-Chemokines induce the directional migration of Monocytes and T-Lymphocytes that are implicated in the PathoGenesis of Multiple Sclerosis (MS) lesions.

RANTES is a member of the beta-Chemokine family that has been detected in the Lesions of MS patients.

However, the cellular sources of RANTES message and the molecular basis for the regulation of its production in MS lesions are not well understood.

Glial Cells may be a major source of RANTES in vivo and have been shown to produce RANTES in vitro.

Thus, the objective of this study was to establish a model system for studying the regulation of RANTES expression by Cytokines in cultured human Glial Cells, and to determine the mechanism involved in the process.

We show that the Th1 Cytokines TNF- and IL-1ß independently induce RANTES mRNA and Chemokine levels in human U-251 MG Astroglial Cells, and that these effects are time- and concentration-dependent.

In addition, we demonstrate that both Cytokines increased the rate of transcription of the RANTES Gene, as estimated by in vitro nuclear transcript elongation assays.

The transcriptional activity in TNF--treated U-251 MG cells started to increase at 2 h and peaked at 8 h, with levels more than 14 times greater than controls.

We further show that NF-kappaB may play a critical role in the up-regulation of human RANTES Gene expression in this system.

Gel shift assays revealed an induction of in vitro nuclear extract binding activity to the NF-kappaB element of RANTES in cells incubated with the Th1 Cytokines.

These observations suggest that human Astroglia, within diseased Brain, may be stimulated to produce RANTES Chemokine in response to TNF- and IL-1ß.

And, that this effect of the Th1 Cytokines is attributed to increase of transcription.