The serotonergic agent m-chlorophenylpiperazine induces migraine attacks: A controlled study.

In a double-blind study versus placebo, the serotonergic agent m-chlorophenylpiperazine (mCPP) was administered to 20 healthy control subjects and 19 migraineurs to investigate the ability of mCPP (0.5 mg/kg) to induce typical migraine attacks. In the following 24 hours there were more migraines after mCPP than after placebo in both groups. These findings are consistent with involvement of 5HT2B,2C,1A receptor subtypes in the pathophysiology of migraine.

Effects of beta-IFN-1b treatment in MS patients on adhesion between PBMNCs, HUVECs and MS-HBECs: an in vivo and in vitro study.

The in vivo effects on the expression of adhesion molecules and on the adhesion between mononuclear cells and multiple sclerosis human brain endothelial cells (MS-HBECs) were investigated at the beginning of beta-IFN-1b treatment of MS patients. MS-HBECs were isolated from a surgical specimen obtained from an MS patient undergoing brain surgery for vascular aneurysm. 48 h after the first single administration of beta-IFN-1b, PBMNCs of 10 MS patients were analyzed for HLA-DR, CD11a, CD18 and VLA-4 expression and the adhesion between PBMNCs and both stimulated and unstimulated MS-HBECs evaluated. sICAM-1 and sVCAM-1 dosage in the serum of the patients was checked as well. The experiments were repeated using HUVECs in order to detect possible endothelial organ-specific differences. The experiments were also performed after six months of beta-INF-1b treatment on HUVECs. No significant effects on mononuclear cells/endothelium adhesion were detected at 48 h, but adhesion of PBMNCs to HUVECs decreased at six months. An increase in HLA-DR and VLA-4 and a decrease of CD18 was detected in monocytes. The serum level of sVCAM-1 increased at T2 and was still higher than at T0 at six months. The effect of the beta-IFN-1b treatment on both MS-HBECs and HUVECs, was selectively studied in vitro by testing the expression of cytokine-induced adhesion molecules HLA-DR, ICAM-1 and VCAM-1. The in vitro experiments confirmed that beta-IFN-1b is able to antagonize gamma-IFN-induced HLA-DR expression on MS human brain endothelial cells without relevant effects on VCAM-1 and ICAM-1.

5-HT1A receptor hypersensitivity in migraine is suggested by the m-chlorophenylpiperazine test.

Involvement of the cerebral serotoninergic system has been invoked to explain the origin of the pain and the vascular phenomena in migraine. To further investigate the type of cerebral serotonin receptors that may be altered in migraine, the prolactin (PRL) and cortisol responses to m-chlorophenylpiperazine (mCPP), a selective 5-HT1A,-5-HT(2A/C) receptor agonist, were monitored in 12 patients suffering from migraine without aura and in 14 matched healthy controls. Each subject underwent two challenges, one with mCPP (0.5 mg/kg) and the other with placebo (orally) using a double-blind crossover design. Anxiety level was measured by the State Trait Anxiety Inventory. Migraine patients had a greater PRL response to mCPP (p = 0.05) and greater anxiety (p < 0.01) than controls; cortisol response to mCPP did not differ suggesting that 5-HT2C receptors are normal in migraine. Augmented PRL response to mCPP could derive from 5-HT1A receptor hypersensitivity, perhaps as as a consequence of anxiety due to pain expectation. Cerebral 5-HT1A hypersensitivity could also explain the increased occurrence of migraine attacks during anxiety.

Immunological monitoring of azathioprine treatment in multiple sclerosis patients.

Despite the longstanding clinical use of azathioprine as an immunosuppressive agent in multiple sclerosis, little is known about the action of this drug on a number of parameters of putative pathogenic relevance in the disease. Eleven patients with multiple sclerosis, treated with azathioprine 2.5-3 mg/kg per day, and six untreated patients were studied with serial blood sampling for 1 year. The following immunological parameters were investigated: peripheral blood lymphocyte subsets, natural killer activity, serum IgG, IgM, ICAM-1 and tumour necrosis factor alpha (TNF-alpha). The most relevant changes included a decrease in CD3- CD56+ cells, an increase in CD4+ CD45RA+ cells and a decrease in TNF-alpha levels only in treated patients, while no changes occurred in untreated patients over a 1-year period. The decrease in TNF-alpha levels and the increase in "suppressor-inducer" lymphocytes could reduce chronic inflammation in multiple sclerosis, and paralleled an overall favourable clinical response to azathioprine treatment in our patients.

HLA and multiple sclerosis in Italy: a review of the literature.

HLA antigens of locus A, C, B, DR and DQ were typed in 104 Italian multiple sclerosis patients and in 905 healthy controls; the results have been compared with those published in the Italian literature. The Italian studies have been reviewed regarding the ethnic origin of the typed population and the corresponding prevalence of the disease. The data suggest a lack of association between A3 and B7 antigens and Italian multiple sclerosis and a relevance of other DR locus antigens (mainly DR4 and DR5), in addition to DR2, in the susceptibility to the disease.

CSF T-cell subsets in multiple sclerosis: relationship to cerebrospinal fluid myelin basic protein and clinical activity.

Cerebrospinal fluid myelin basic protein and cerebrospinal fluid and peripheral blood T-cell subsets have been studied in patients with multiple sclerosis and other inflammatory and non-inflammatory nervous system diseases. These biological parameters have been correlated with clinical disease activity. No changes in peripheral blood T-cell subsets were seen in multiple sclerosis patients. Low cerebrospinal fluid T8+ cells occurred only in multiple sclerosis, while high cerebrospinal fluid T4+ cells were detected both in clinically active multiple sclerosis and in inflammatory nervous system diseases. A close relationship was found between cerebrospinal fluid T4/T8 ratio and myelin basic protein in relapsing multiple sclerosis patients.

Cryptococcal meningoencephalitis: intrathecal immunological response.

The intrathecal immune response is reported in a patient with cryptococcal meningoencephalitis. CSF IgM and IgG levels were significantly related to the favourable clinical evolution. IgM response was specifically directed against the pathological agent, while IgG were mostly non-specific. The data are discussed and compared with the other chronic infections of the central nervous system.

Low serum interleukin-10 levels in multiple sclerosis: further evidence for decreased systemic immunosuppression?

Serum interleukin 10 (IL10) levels were assessed in patients with multiple sclerosis who were either in a stable or active clinical condition. The levels were compared with values in healthy controls. Lower IL10 levels than in controls were seen in multiple sclerosis patients, regardless of clinical disease activity. Low IL10 levels were also seen in patients with systemic lupus erythematosus. No clear-cut relationships emerged between IL10 levels and those of tumour necrosis factor alpha and transforming growth factor beta, or between IL10 and lymphocyte subsets in peripheral blood.

Immunological effects of in vivo interferon-beta1b treatment in ten patients with multiple sclerosis: a 1-year follow-up.

Ten patients with multiple sclerosis and treated with interferon-beta1b (IFN-beta1b) were followed-up for 1 year with quantitation of serum VCAM-1 and ICAM-1 levels, mean fluorescence intensity of HLA-DR, VLA-4, CD11a, and CD18 on peripheral blood monocytes and lymphocytes, and adhesion of peripheral blood monocytes and CD45+ cells on endothelial cell monolayers. Adhesion molecule expression and adhesion of peripheral blood monocytes to endothelium were also monitored in healthy controls. No differences in adhesion were detected between MS patients before treatment and healthy controls, while after 1 year a marked decrease in the number of monocytes and mononuclear cells adhering to human umbilical vein endothelial cell monolayers was observed in patients treated with IFN-beta1b. After 1 year of treatment a significant increase in HLA-DR on peripheral blood monocytes was also detected. Our findings regarding lowered adhesion add information to available evidence of the mechanisms of action of IFN-beta1b in MS.

A dry-reagent strip for quantifying carbamazepine evaluated.

We examined a new colorimetric homogeneous immunoassay for carbamazepine based on the apoenzyme reactivation immunoassay system (ARIS) principle. The test, in dry-reagent strip format, is to be used with the Ames Seralyzer reflectance photometer. Within-run CVs (n = 20) were 3.0%, 2.7%, and 2.8% at 3.0, 6.1, and 12.1 mg/L; between-run CVs (n = 15, in 15 days) were 4.1%, 2.7%, and 1.9% at 6.0, 9.1, and 12.1 mg/L. Mean analytical recovery was 99.9 (SD 2.3)%. Results by this test (y) for clinical plasma specimens (n = 96) compared very well with those obtained by fluorescence polarization immunoassay (y = 1.01 x - 0.02, r = 0.995) and by liquid chromatography (y = 0.99 x + 0.14, r = 0.990). Bilirubin (45 mg/L), uric acid (145 mg/L), and various anticoagulants at about fourfold the usual concentrations did not interfere. High concentrations of cholesterol (4.9 g/L), triglycerides (3.8 g/L), and hemoglobin (4 g/L) caused slight positive interference. Carbamazepine-10,11-epoxide cross reacted only at greater than or equal to 5 mg/L. The two-point calibration line was validly stored for at least three weeks. Free carbamazepine also can be measured. The test is convenient and rapid (test time 80 s), and thus is particularly useful in all clinical settings where prompt testing is needed.

Quantitative determination of phenobarbital and phenytoin by dry-phase apoenzyme reactivation immunoassay system (ARIS).

We assessed the performance of the apoenzyme reactivation immunoassay system (ARIS) reagent strip tests for determination of phenobarbital (PB) and phenytoin (PHT) with the Seralyzer reflectance photometer. In the assay, the drug of the sample competes with a flavine adenine dinucleotide (FAD)-drug conjugate for binding to a specific antibody; the unbound conjugate then activates apoglucose oxidase to reconstitute glucose oxidase, whose activity is kinetically monitored by a coupled chromogenic reaction. Within-run coefficients of variation (CVs) were less than or equal to 5.0% for PB and less than or equal to 5.6% for PHT; between-run CVs were less than or equal to 6.1% for PB and less than or equal to 6.5% for PHT. Mean analytical recoveries were 100.3% for PB and 100.2% for PHT. Test results were not significantly affected by bilirubin (5 mg/dL), hemoglobin (25 mg/dL), triglycerides (500 mg/dL), uric acid (15 mg/dL), or elevated levels of other antiepileptic drugs. Reagent strip tests correlated very well with substrate-labeled fluorescent immunoassay (r = 0.9923 and 0.9944 for PB and PHT, respectively), enzyme multiplied immunoassay technique (r = 0.9941 and 0.9919), and gas-liquid chromatography (r = 0.9980 and 0.9960). These homogeneous competitive colorimetric immunoassays are particularly suitable for emergency use, for testing small batches of samples, wherever prompt results are needed.

A simple immunoturbidimetric method for IgG and albumin quantitation in cerebrospinal fluid and serum.

We describe a simple immunoturbidimetric method for measuring both IgG and albumin in CSF and serum, which enables the calculation of CSF indices. For each protein, only one calibration curve is used for both CSF and serum samples. The assay protocol is simple and similar for both tests. Sensitivity and versatility of the method afford measurements over a very wide range of concentrations (approx. 0.007 to 94 g/l for IgG and 0.06 to 92.40 g/l for albumin). Precision studies (triplicates for 6 runs over 15 days) gave overall CVs: less than or equal to 2.9 and 4.9% for IgG in CSF (11.5 mg/l) and serum (10.28 g/l); less than or equal to 1.3 and 1.1% for albumin in CSF (115 mg/l) and serum (76.89 g/l). Comparison studies showed good correlation with radial immuno-diffusion (r greater than or equal to 0.995 and 0.976 for IgG and albumin) and rate nephelometry (r greater than or equal to 0.967 and 0.982 for IgG and albumin). Thus, the method under investigation proved to be reliable and appears to be particularly suitable for the routine work.

CFS oligoclonal bands in MS and other neurological diseases detected by agarose isoelectric focusing and electrophoresis.

Agarose gel isoelectric focusing (IEF) and electrophoresis (EF) were compared for detection of CSF oligoclonal bands in multiple sclerosis (MS) and other neurological diseases. The CSF IgG/albumin ratio, the IgG-Index and the IgG synthesis per day in the CNS were also considered. IEF was highly sensitive, revealing oligoclonal bands in 100% of CSF from 40 clinically definite MS, while EF had a sensitivity of 70%. In 90 patients with other neurological diseases, IEF revealed oligoclonal bands in 25.5%, EF in 11.1%. The IgG-index was the most sensitive of the quantitative parameters.

Immunological alterations in cluster headache during remission and cluster period. Comparison with low back pain patients.

Cluster headache is a disorder of unknown origin. Some studies have focused their attention on neuroendocrine derangement, others on immunity. To probe central alterations in cluster headache (CH), immune parameters were investigated in cluster headache patients in comparison to low back pain patients and healthy controls. Increases in peripheral blood monocytes found in remission cluster headache patients may be attributable to chronic central nervous system (hypothalamic?) noradrenergic dysfunction or altered beta-endorphin. Alterations in NK+, CD3+ and CD4+ levels found in cluster period cluster headache and low back pain patients are probably pain or stress-related.

Analysis of peripheral blood lymphocyte phenotype and function during dexamethazone treatment of progressive multiple sclerosis.

Five patients with chronic progressive multiple sclerosis (MS) and three control patients with lumbar disc herniation were treated with dexamethazone during 14 days. The effect on peripheral blood T-cell subsets and on the proliferative response of peripheral blood mononuclear cells (PBMC) to pokeweed mitogen (PWM) and anti-mu antibody was analyzed. Before treatment, the proportion of CD3+ and CD4+ PBMC was similar in MS and control patients, but the proportion of CD8+ and DR+ PBMC was lower and the PBMC were less responsive to anti-mu stimulation in MS patients compared to controls. Steroid treatment induced reversible granulocytosis and lymphocytosis. CD3+ and CD4+ cells increased and DR+ cells decreased in MS patients but not in controls. Proliferation of anti-mu stimulated PBMC increased in MS-patients during the two weeks of treatment, but decreased in controls. The enhancement in the MS patients of pre-existing immune abnormalities suggests that a cautious attitude is warranted in the use of steroid treatment in chronic progressive MS.