Association of polymorphisms of the cannabinoid receptor (CNR1) and fatty acid amide hydrolase (FAAH) genes with heroin addiction: impact of long repeats of CNR1.

Alterations in expression of a cannabinoid receptor (CNR1, CB1), and of fatty acid amide hydrolase (FAAH) that degrades endogenous ligands of CB1, may contribute to the development of addiction. The 385C>A in the FAAH gene and six polymorphisms of CNR1 were genotyped in former heroin addicts and control subjects (247 Caucasians, 161 Hispanics, 179 African Americans and 19 Asians). In Caucasians, long repeats (>or=14) of 18087-18131(TAA)(8-17) were associated with heroin addiction (P=0.0102). Across three ethnicities combined, a highly significant association of long repeats with heroin addiction was found (z=3.322, P=0.0009). Point-wise significant associations of allele 1359A (P=0.006) and genotype 1359AA (P=0.034) with protection from heroin addiction were found in Caucasians. Also in Caucasians, the genotype pattern, 1359G>A and -6274A>T, was significantly associated with heroin addiction experiment wise (P=0.0244). No association of FAAH 385C>A with heroin addiction was found in any group studied.

The islands of Calleja: organization and connections.

The islands of Calleja (IC) in the rate are composed of seven small groups of granule cells in the polymorph layer of the olfactory tubercle and one large group, the insula magna, which lies along the border between septum, nucleus accumbens and nucleus of the diagonal band. The cytoarchitecture and neuronal morphology of the IC and surrounding cells, studied using Nissl-stained and Golgi-Kopsch material, are described. In addition, the afferent and efferent connections of the IC were analyzed using fluorescence histochemistry, the autoradiographic tracing method, and the anterograde and retrograde horseradish peroxidase methods. Topographically organized projections to the IC from the dopamine-containing cells of the substantia nigra-ventral tegmental area are demonstrated by the glyoxylic acid fluorescence histochemical method and the autoradiographic tracing technique. Anterograde and retrograde horseradish peroxidase studies provide evidence for reciprocal, topographically organized interconnections between the IC and the septum, nucleus accumbens, amygdala and piriform cortex. These observations indicate that the IC constitute a unique population of granule cells, located in the olfactory tubercle, innervated by dopamine neurons of the mesencephalon and interconnected with olfactory and non-olfactory components of the basal forebrain.

Primary intracranial hypotension and bilateral isodense subdural hematomas.

A 39-year-old woman with headache and an organic mental syndrome was found to have primary intracranial hypotension (PIH). Bilateral isodense subdural hematomas were discovered in association with an absence of detectable CSF pressure on two lumbar punctures. This case study emphasizes that PIH is not an entirely benign condition and that intracranial hemorrhage may accompany persistent intracranial hypotension.

A neurologic rating scale (NRS) for use in multiple sclerosis.

A neurologic rating scale (NRS) has been developed for clinical assessment of MS patients. The scale has been tested on 250 MS patients. Assignment of the NRS score is based on assessment of each component of the neurologic examination and accurately reflects overall neurologic function. Clinical exacerbations are evident as significant deviations from baseline scores. There was close interexaminer correlation, with the range of variability no greater than 2.6%. The NRS is a simple, reliable, and sensitive scale that can be used with other objective measurements of neurologic function, such as neurophysiologic studies, in the clinical assessment of MS patients.

Systemic alpha-interferon therapy of multiple sclerosis.

A randomized, double-blind, placebo-controlled crossover study tested the efficacy of natural alpha interferon in altering exacerbating-remitting MS. Twenty-four patients with frequent exacerbations were treated for 6-month periods, beginning with either 5 X 10(6) IU of interferon daily or placebo. A 6-month washout period followed each treatment. Exacerbation rates were reduced during interferon and placebo phases compared with pre-study rates; a greater reduction occurred on interferon, particularly following placebo, possibly reflecting a learning phenomenon. Fifteen patients with a strictly exacerbating-remitting course had fewer and milder exacerbations on interferon compared with those on placebo, whereas 9 patients with a progressive component continued to have active disease. These results suggest that interferon might reduce exacerbations in certain patients and indicate guidelines for future trials of interferon in MS.

Disease activity markers in multiple sclerosis. Another look at suppressor cells defined by monoclonal antibodies OKT4, OKT5, and OKT8.

Here we report our experience in profiling peripheral blood T-cell subsets with the monoclonal antibodies OKT4, OKT5 , and OKT8. Lymphocyte surface phenotype was measured by automated cytofluorometry. In a population survey, we were unable to detect differences between patients with multiple sclerosis (MS) and control subjects when we compared ratios of lymphocytes of helper cell phenotype (OKT4) to those with suppressor cell phenotype ( OKT5 and OKT8). No differences could be established between patients with stable disease, chronic progressive disease, or those with active disease. In a study of 10 patients followed through an exacerbation, we were also unable to define perturbations in these lymphocyte ratios that correlated with disease activity. Detailed analysis of the fluorescence histogram, which examines the entire spectrum of cell surface fluorescence intensity in a population of lymphocytes, was also not useful in predicting disease activity in these patients. The discrepancies between these data and other reports in the literature are discussed. We propose that these reagents are inadequate indices of disease activity, and that until other monoclonal reagents are developed and studied, the suppressor cell compartment is best assessed by assays of function.

Cladribine.

Cladribine is a novel drug that selectively depletes lymphocytes and may be able to destroy the activated immunocytes that damage the central nervous system in multiple sclerosis. Our initial controlled studies have shown a beneficial, although temporary, dose-related effect of cladribine on the course of chronic progressive multiple sclerosis. Peak improvement in median Scripps Neurological Rating Scale (SNRS) neurological performance scores, followed by gradual decline, occurred at month 14 after initiation of treatment with a 2.8 mg/kg total dose and at month 7 after initiation of treatment with a 1.4 mg/kg total dose. A marked decrease in the presence of enhanced magnetic resonance imaging lesions was observed at both dose levels. Adverse effects are also dose-related. Mild segmental herpes zoster or transient marrow suppression occurred in some patients treated at the higher total dose, whereas no problems of this kind were observed at the lower total dose. It is our hope that studies that are presently under way will establish cladribine as a practical therapeutic option for patients with all non-benign forms of multiple sclerosis.

A comparison of two neurologic scoring instruments for multiple sclerosis.

We examined the degree of association between two neurologic impairment scales, the Extended Disability Status Scale (EDSS) and the Scripps Neurologic Rating Scale (SNRS), with data from a randomized, double-blind, placebo-controlled clinical trial assessing the safety and efficacy of cladribine as treatment for chronic progressive multiple sclerosis (MS). We found that the EDSS and SNRS were not strongly correlated within individual patients, contrary to expectation; moreover, in 9 of the 48 evaluable patients, the directions of their changes from baseline values were not mutually consistent. The scales were differentially sensitive to clinical changes over time, with the EDSS indicating a more abrupt, and the SNRS a more gradual, change in the clinical course of disease. The validity of different impairment scales, and their sensitivity to detect clinical changes, should be formally assessed in future clinical trials using these scales as outcome measures.

Predictive value of lesions for relapses in relapsing-remitting multiple sclerosis.

BACKGROUND AND PURPOSE: Recent studies have suggested that enhancing lesions on contrast-enhanced T1-weighted MR images are predictive of impending exacerbations in cases of relapsing-remitting multiple sclerosis. We examined whether enhancing lesions, new enhancing lesions, and new hypointense lesions ("black holes") could accurately predict exacerbations in a cohort of 50 patients with relapsing-remitting multiple sclerosis within a time frame of up to 6 months. METHODS: Data were obtained from 50 patients with relapsing-remitting disease. All patients underwent monthly MR imaging and clinical examinations for a period of 12 months. Putative predictors of clinical relapse were defined from enhancing lesions, new enhancing lesions, and new black hole outcomes, and their operating characteristics were studied. RESULTS: Overall, the positive predictive values (PV+) of enhancing lesions, new enhancing lesions, or new black holes for an exacerbation did not exceed 0.25 and the negative predictive values (PV-) were all near 0.9. The best predictor for new enhancing lesions was the occurrence of new enhancing lesions in each of the previous 3 months (PV+: 0.79 [95% confidence interval, 0.651-0.900]; PV-: 0.83 [95% confidence interval, 0.751-0.887]). Similarly, new black holes were predicted best by the occurrence of new black holes in each of the previous 2 months (PV+: 0.54 [95% confidence interval: 0.372-0.697]; PV-: 0.85 [95% confidence interval, 0.790-0.896]). CONCLUSION: None of the MR markers could predict an impending relapse with any reasonable degree of precision. Rather, the absence of MR markers is associated with a more favorable clinical course (ie, fewer relapses).

Method for intracellular magnetic labeling of human mononuclear cells using approved iron contrast agents.

A method for intracellular iron labeling of human mononuclear cells (lymphocytes and monocytes) for magnetic resonance imaging (MRI) using simple incubation of cells with approved MRI iron contrast agents is presented. Labeled cells can be detected by MRI in vitro, and this suggests the possibility that the technique could become a marker for in vivo lymphocyte and monocyte trafficking studies in acute inflammatory lesions such as those in Multiple Sclerosis.

Responsiveness of the Scripps neurologic rating scale during a multiple sclerosis clinical trial.

OBJECTIVE: The Scripps neurologic rating scale (SNRS) is a summary measure of individual components comprising a neurological examination, designed for use in multiple sclerosis (MS). Our objective is to evaluate the responsiveness of the SNRS, within the context of a 2-year, randomized, double-blind crossover study of the efficacy of cladribine for treatment of secondary progressive MS. METHODS: Effect sizes were determined for the SNRS and its components, separately for each treatment group (initial placebo, and initial cladribine) over both years of the clinical trial, using a standard random effects model. RESULTS: Individual components tended to show positive effect sizes (improvement) during periods of active therapy in both treatment groups, and negative effect sizes (deterioration) during periods of no active therapy. Summation indices derived from the individual components of the SNRS seemed somewhat more stable than the individual components. The two components mentation and mood, and bladder, bowel, or sexual dysfunction, were rather unresponsive in our clinical trial. CONCLUSIONS: Changes in the components of the SNRS over the course of our clinical trial were consistent between the two treatment groups. Most components were moderately responsive; and, the summary SNRS score appropriately summarized the moderate magnitudes of change evinced in the individual components.

P300 in multiple sclerosis: a preliminary report.

The P300 component of the event-related brain potential (ERP) elicited with auditory stimuli and pattern-shift visual evoked potentials (VEPs) was obtained from 16 patients with multiple sclerosis (MS) and 16 matched control subjects. P300 latency was significantly longer and component amplitude relatively depressed in the MS patients compared to control subjects. The P100 potential of the VEP also was delayed for both full-field and half-field stimulus conditions in the patients compared to control subjects. The findings suggest that the P300 ERP may reflect the cognitive decline associated with MS.

Overweight and obesity associated with a missense polymorphism in fatty acid amide hydrolase (FAAH).

BACKGROUND: The brain endogenous cannabinoid system modulates reward and craving pathways and consequently may affect body weight. A naturally occurring missense polymorphism in the gene encoding fatty acid amide hydrolase (FAAH), the primary enzyme for inactivation of endocannabinoids, is associated with problem drug use. AIMS: To investigate the relationship between the FAAH cDNA 385 A/A (P129T) polymorphism and overweight disorders in subjects of multiple ethnic backgrounds attending a medical screening clinic. SUBJECTS: A total of 2667 subjects of white, black and Asian ancestry were genotyped and stratified by a standardized clinic-based assessment of body mass index (BMI, weight in kilograms/(height in meters)(2) or kg/m(2)). METHODS: Subjects were genotyped for the FAAH cDNA 385 C --> A polymorphism using allele-specific oligonucleotide hybridization methods by investigators blinded to all clinical information. BMI was calculated based on exact clinical measurements and World Health Organization ranges were used to stratify subjects. Statistical methods included the Fisher exact test, Mann-Whitney U-test and multivariable logistic regression analysis. RESULTS: The homozygous FAAH 385 A/A genotype was significantly associated with overweight and obesity in white subjects (P=0.005) and in black subjects (P=0.05) but not in a small group of Asians. The median BMI for all subjects was significantly greater in the FAAH 385 A/A genotype group compared to heterozygote and wild-type groups (P=0.0001). In white subjects, there was an increasing frequency of the FAAH 385 A/A genotype with increasing BMI categories of overweight (P=0.02) and obese (P=0.006) with the same trend in black subjects. CONCLUSIONS: These results suggest a role for the FAAH 385 A/A missense polymorphism as an endocannabinoid risk factor in overweight/obesity and may provide indirect evidence to support cannabinoid antagonist treatment strategies in overweight disorders.

Gap junctions between astrocytes during growth and differentiation in organ culture systems.

Fetal rat neocortex maintained in organ culture systems with the use of sponge foam matrices and millipore filter platforms undergoes growth and cytodifferentiation along classical neuronal and glial lines up to 36 days in vitro (DIV). Astrocytic differentiation is characterized by accumulation of 80-90 A glial filaments in the cell bodies and processes of astrocytes. Gap or nexus junctions closely resembling those formed in mammalian brain in situ are identified by 15 DIV. By 36 DIV, interastrocytic gap junctions are numerous and frequently join extensive lengths of adjacent glial plasma membranes. The results suggest that these organ culture systems may provide a favorable environment for the study of cellular structure and function of coupled neuroglia.