Induction of arginase activity with the Shope papilloma virus in tissue culture cells from an argininemic patient.

Inoculation of the Shope virus in tissue cultures of human fibroblasts from a patient with a deficiency of the enzyme arginase results in an induction of arginase activity, apparently virus coded.

Antiplatelet therapy is effective in primary prevention of myocardial infarction in patients with a previous cerebrovascular ischemic event.

OBJECTIVE: A secondary subgroup analysis of the European Stroke Prevention Study of the effect of antiplatelet medication on the risk of myocardial infarction. DESIGN AND SETTING: A randomized, double-blind placebo-controlled study with two parallel treatment groups (dipyridamole plus aspirin and placebo). Sixteen centers from six countries participated in the study. PATIENTS: A total of 2500 patients who had had one or more transient ischemic attacks or cerebral infarctions participated. INTERVENTION: Combination therapy with dipyridamole (75 mg three times a day) and aspirin (330 mg three times a day) was compared with placebo during 24 months' follow-up. OUTCOME MEASURES: Prevention of fatal and nonfatal myocardial infarction. RESULTS: A total of 105 myocardial infarctions occurred in the intention-to-treat analysis and 76 occurred in the explanatory analysis. The overall risk reduction of myocardial infarction with the study drugs was approximately 40% in both statistical analyses, but the result was statistically significant only in the intention-to-treat analysis. Therapeutic efficacy was better among male patients, patients younger than 65 years, and patients with hypertension. CONCLUSION: Combination therapy with dipyridamole and aspirin reduces not only the risk of cerebrovascular ischemic events but also the risk of myocardial infarction.

The chemical composition of idiopathic nonarteriosclerotic cerebral calcifications.

Calculi from a case of cerebral idiopathic nonarteriosclerotic calcification (Fahr's disease) were examined. The stone consists of hydroxyapatite and possesses a typical structure: the calcification process seems to be initiated by the formation of small round bodies that are cemented to each other to form the final stone. Calcified vessels are also present, but seem to be a secondary effect. From a comparison with other calcifications, it is concluded that no pathologic significance should be attached to the relatively high levels of trace metals such as zinc, iron, copper, magnesium, lead, and others, with the possible exception of manganese. The organic matrix of the stone contains large quantities of protein. On hydrolysis of this fraction, an important unidentified ninhydrin-positive peak was found. No mucopolysaccharides were found.

Arachnoiditis ossificans with arachnoid cyst after cranial tuberculous meningitis.

A 34-year-old woman developed symptomatic arachnoiditis ossificans and an arachnoid cyst as a consequence of tuberculous meningitis adequately treated 20 years before. Surgical decompression of the cyst stopped the progression of her spastic paraparesis. Pathologic examination confirmed the presence of ossification of the arachnoid.

The European Stroke Prevention Study: results according to sex.

The European Stroke Prevention Study was a multicenter trial comparing the effect of a combination of 75 mg dipyridamole and 330 mg acetylsalicylic acid tid with placebo in the prevention of stroke or death after one or more attacks of recent transient ischemic attacks or stroke of atherothrombotic origin. From the 2,500 patients in the intention-to-treat analysis, the proportion of women was 42%, and from the 1,861 patients in the explanatory analysis it was 44%. The endpoint incidence was significantly higher in men than in women. The endpoint reduction was statistically significant only in the intention-to-treat analysis with total endpoints. However, there was a marked percentage reduction of endpoints in both men and women in explanatory analysis. The risk reduction of strokes was 49% for men and 41% for women, and the reduction of total endpoints was 39% in men and 30% in women. Thus, antiplatelet therapy is effective in the prevention of stroke or death in both sexes.

DGAVP (Org 5667) in early Alzheimer's disease patients: an international double-blind, placebo-controlled, multicenter trial.

We carried out a double-blind study of a vasopressin-related peptide, DGAVP citrate (Org 5667), in 115 patients with mild dementia, probable Alzheimer's type (DAT). Neither clinical rating scales nor psychometric tests revealed any improvement over 84 days with once-daily intranasal treatment with 2 different doses of DGAVP. We conclude that vasopressin-like peptides are not satisfactory therapeutic agents in DAT.

Antiplatelet treatment does not reduce the severity of subsequent stroke. European Stroke Prevention Study 2 Working Group.

OBJECTIVE: To assess the the effect of antiplatelet therapy on the severity of subsequent stroke in patients with stroke and TIA. BACKGROUND: The Second European Stroke Prevention Study (ESPS2) recruited 6,602 patients in four treatment groups: placebo, 2 x 25 mg acetylsalicylic acid (ASA), 2 x 200 mg dipyridamole (DP), and the combination of 50 mg ASA and 400 mg DP per day. Seventy-six percent of the patients had had a stroke as the qualifying event, whereas 24% had a TIA. All patients were followed at 3-month intervals for 2 years. ESPS2 showed a benefit from antiplatelet treatment compared with placebo and an additional benefit using ASA and DP together compared with either of these antiplatelet agents alone. METHODS: In the ESPS2, the study protocol included assessment of severity of end point stroke with the modified Rankin scale once the stroke had clinically stabilized, and no further impairment was observed. There were 824 new stroke events during follow-up. In 701 of them, the initial Rankin scale was known, and this was also evaluated after each nonfatal recurrent stroke. The difference in Rankin scale between treatment groups was analyzed after recurrent stroke, and the progress in Rankin scale between entry and recurrent stroke was quantified by calculating the number of patients with a change of one or more degrees in the scale. RESULTS: There were no significant differences in these changes in Rankin scale between the treatment groups. The mean time to reach an end point of stroke was longest in patients who used ASA + DP (p = 0.057). However, there was no difference among the treatment groups in the time to death during follow-up. CONCLUSION: This study suggests that antiplatelet therapy does not influence the severity of recurrent stroke as evaluated with the Rankin scale. However, antiplatelet therapy seems to lengthen the time the patient remains free from a recurrent stroke.

Serological identification of viral antigens after electrophoretic transfer.

The method described permits detection of specific antibodies to antigenic material, the protein components of which can first be separated by electrophoresis and then transferred to a supporting medium where their reactivity with antibody may be demonstrated using peroxidase-labelled anti-Ig. Results are described for measles virus and measles-like virus isolated from subacute sclerosing panencephalitis (SSPE) brain. Dilutions, incubation time and conditions, detection with peroxidase-labelled immunoglobulins and selection of substrate are described and discussed.

A two-site immunoradiometric assay for the determination of alpha-albumin.

A two-site immunoradiometric assay for a brain specific protein (alpha-albumin or glial fibrillary acidic protein) is described which may be applied to other proteins. The assay variables studied are immunoadsorbent dilution, temperature, incubation times, reactivity of the labelled antibodies and effect of the washing. Each of these factors is optimalised, resulting in a sensitive, specific, reliable assay. The working range is 5--5000 ng/ml, the within-assay variance is 5%. No high dose hook effect is seen in this range. The method is applied for the measurement of alpha-albumin present in cerebrospinal fluid, nervous tissue extracts and cell cultures.

Antibodies-restricted heterogeneity in serum and cerebrospinal fluid of chronic relapsing experimental allergic encephalomyelitis.

An animal model which might help to study multiple sclerosis has long been sought. With chronic relapsing experimental allergic encephalitis (EAE), the search seems to have brought hope and evidence of comparable pathology whether concerned with clinical or neuropathological results, but no study of the cerebrospinal fluid (CSF) electrophoretic pattern has been made so far. A new sensitive method enables to study the CSF proteins: unconcentrated CSF proteins after agar gel electrophoresis are stained with silver reagents. The silver technique allows to follow the evolution of the inflammatory reaction in chronic relapsing EAE as well as in the acute form of EAE. This technique provides an additional approach to the study of EAE and an argument in favor of chronic relapsing EAE in guinea pigs as a model for multiple sclerosis.

Primary cultures from defined brain areas; effects of seeding time on cell growth, astroglial content and protein synthesis.

The influence of seeding time on cell growth, astroglial content and on protein synthesis during cultivation was determined in primary cultures from 3 phylogenetically different brain areas from rat cerebral cortex, striatum and brainstem. Brainstem cultivated from 17-day-old embryos and all the cultures studied from the 3 brain areas of newborn and 7-day-old rat showed a similar increase in total and water-soluble protein during cultivation. Glial fibrillary acidic protein (GFAp, alpha-albumin) levels increased with age in all cultures studied. There was a rapid increase in GFAp (alpha-albumin) between 1 and 2 weeks in cultures from newborn and between 2 and 3 weeks in brainstem cultures from 17-day-old embryos, these increases being slower thereafter. Incorporation of [3H]valine into soluble protein was lower in 3-week-old cultures than in 1- and 2-week-old cultures derived from newborn and 7-day-old rat brain. The incorporation rates were similar in comparisons of the various cultures. Similar results were obtained from embryonic cultures, although the decrease in incorporation rate was between 3 and 4 weeks. The efficiency of incorporation (% TCA-precipitated material/total [3H]activity) was higher in 2- and 3-week-old than in 1-week-old cultures from newborn and 7-day-old rats and in 3- and 4-week-old cultures of brainstem from 17-day-old rat embryos. These findings suggest a cell differentiation during cultivation. The results show that seeding time has a variable influence on cultures from the different brain areas studied concerning cell growth, astroglial content and probably differentiation during cultivation. Embryonic cell cultures seem, in general, to develop one week later than neonatal and postnatal ones. Cultures of newborn rat cells from cerebral cortex, striatum and brainstem show many similarities in the above parameters during cultivation. This is also the case for brainstem cultures from embryonic rat.

Multiple sclerosis: oligoclonal IgG, kappa-lambda light chain distribution and measles antibodies in brain extracts.

The presence of measles antibodies in white and grey brain material and in 8 demyelination plaques from 6 patients affected with multiple sclerosis was investigated with the hemagglutination inhibition (HI) and complement fixation (CF) techniques. White and grey matter of 5 controls were run in parallel. No measles antibodies could be detected, except for one plaque, where the answer could be considered as doubtful. The immunoglobulin G (IgG) content and the kappa/lambda light chain ratios of all the samples were evaluated. No unique monoclonal immunoglobulin population could be detected, but kappa or lambda predominant oligoclonal populations appeared in controls as well as in MS IgG.

Brain primary culture-- a characterization (part II).

Brain primary cultures from newborn rat hemispheres have been further characterized. THe cultures contained the glial fibrillary acidic protein (GFAp; alpha-albumin), as analyzed quantitatively. GFAp (alpha-albumin) increased in concentration during growth of cultures and during maturation of the intact rat brain. FITC-labelled antibodies against GFAp (alpha-albumin) reacted specifically with astrocyte-like cells. Theses cells were also positive after Rio-Hortega's silver carbonate staining for astrocytes. No myelin basic protein (MBP) was detected in cultures, neither by quantitative measurements nor by immunofluorescent histochemistry. Negative results were also obtained for the myelin specific enzyme 2', 3'-cyclic nucleotide 3'-phosphohydrolase (CNP), e. e. the enzyme was at the same level in the cultures of different ages as in 3-day-old rat hemispheres. However, in rat brain increasing concentrations of MBP and CNP were found during maturation. This means that no myelin and thereby probably no or few differentiated oligodendroglial cells were present in the cultures, where the predominate cell type was of astroglial nature.

Cellular composition of primary cultures from cerebral cortex, striatum, hippocampus, brainstem and cerebellum.

Primary cultures from newborn rat cerebral cortex, striatum, hippocampus, brainstem and cerebellum were grown for 14 days. There was a linear relationship between the amount of material seeded and the protein content of the respective culture. The amount of tissue material seeded was selected so that the different cultures reached confluence at 6-7 days and contained similar amounts of protein when 7 and 14 days old. The cellular content was evaluated by astroglial markers, such as the glial fibrillary acidic protein (GFAp; alpha-albumin) and the S-100 protein, and by markers for other cells expected to be in the cultures (14-3-2 protein, macrophage acidic protein (MAP), alkaline phosphatase, myelin basic protein (MBP), 2',3'-cyclic nucleotide 3'-phosphohydrolase (CNP]. Astroglial-like cells represented 60-70% of the cells present in the different cultures. Quantitation of GFAp (alpha-albumin) showed similar amounts to be present in cultures from cerebral cortex, hippocampus and striatum; however, on lower levels expressed in soluble proteins than in the corresponding brain regions of adult rats. Brainstem of adult rat contained large amounts of GFAp (alpha-albumin), while low levels were found in brainstem culture. Also, phagocytic cells (macrophages), endothelial-like cells, mesenchymal-like cells, ependymal-like cells and oligoblasts were found. Neither mature neurons, nor oligodendroglial cells were observed. It is concluded that although there might be some differences in the degree of maturation or in the cellular composition of the various cultures, they could serve as a good model system for studying the characteristics of astroglial cells from various brain regions.

Diagnostic value of brief microzone electrophoresis of unconcentrated CSF and agar gel electrophoresis of concentrated and unconcentrated CSF.

This paper reports a comparison between brief microzone electrophoresis and agar gel electrophoresis of CSF proteins; 41 different CSFs were examined simultaneously by all three methods. Fractionation of gamma-globulins was found by brief microzone electrophoresis in 53.7% of unconcentrated CSF. This fractionation was found in 63.4% of unconcentrated CSF after agar gel electrophoresis, in 48.8% of concentrated CSF. Serum-like pherogram and the rapid alpha-globulin type were found equally often.

A heat stable serum inhibitor of an antigen antibody reaction of subacute sclerosing panencephalitis.

A factor which inhibits the antigen antibody reaction was isolated from the sera of patients affected with subacute sclerosing panencephalitis (SSPE), multiple sclerosis (MS) and also of control subjects. This factor inhibits the immune reaction between SSPE serum IgGs and measles virus. This inhibiting factor was detected in the alpha globulin region after agar gel electrophoresis, and is heat stable at 100 degrees C for ten minutes.

Biochemical findings in multiple sclerosis. III. Immunoglobulins of restricted heterogeneity and light chain distribution in cerebrospinal fluid of patients with multiple sclerosis.

Several biochemical parameters, the most important of which are total IgG, kappa and lambda light chain distribution and thin layer isoelectric focusing, were determined in the cerebrospinal fluid of 36 multiple sclerosis patients. Their ages and the evolution period of their disease are widely spread and no sex differentiation was made.

Increased GFAp levels in CSF as a marker of organicity in patients with Alzheimer's disease and other types of irreversible chronic organic brain syndrome.

Glial fibrillary acidic protein (GFAp), an astrocyte-specific protein, was determined in cerebrospinal fluid (CSF) of adults and children with global cognitive dysfunction. In children CSF-GFAp values were not closely associated with organic brain disease. However, GFAp values in CSF were increased in 65 of 121 samples of adults with dementia, independent of its cause. GFAp values were not correlated with the severity of the dementia. Increased levels in the CSF are believed to indicate reactive gliosis in most patients with dementia, whereas GFAp levels in encephalitic patients normalize after clinical recovery.

alpha-Albumin (glial fibrillary acidic protein) in normal and pathological human brain and cerebrospinal fluid.

alpha-Albumin, a specific brain protein observed after agar gel electrophoresis and shown to be identical to the later described GFA, has been determined in normal and pathological human central nervous system and cerebrospinal fluid. The outcome of this study underlines the value of the detection of specific proteins of the brain in biological fluids.

Subacute sclerosing panencephalitis antibodies against measles virus polypeptides.

Antibodies to measles virus proteins in SSPE, MS and control human sera were compared using measles virus strain Edmonston, productive SSPE strain Mantooth and non-productive SSPE strain DR. The viral antigens were subjected to transfer electrophoresis, incubated with sera and localized after a second incubation with peroxidase labeled immunoglobulins. High levels of antibodies to all measles viral proteins are present in SSPE sera and CSF and to a lesser degree in MS and control sera although the non-productive SSPE strains lack one viral protein (M protein) present in wild type measles virus strains and the productive SSPE strains.