Massive verapamil overdose complicated by noncardiogenic pulmonary edema.

We describe two cases of pulmonary edema, bradycardia, and hypotension associated with massive verapamil overdose. A noncardiogenic etiology of the pulmonary edema was indicated in one patient by normal thermodilution cardiac output and pulmonary artery occlusion pressure, and in the other patient by a normal echocardiogram. We hypothesize that calcium channel blocker overdose predisposes patients to develop pulmonary edema.

Reversibility of nerve growth factor's enhancement of choline acetyltransferase activity in cultured embryonic rat septum.

Choline acetyltransferase (ChAT) activity and survival of acetylcholinesterase (AChE)-positive neurons were measured in low-density cultures of embryonic (Day 14-15) rat septum exposed to various sequences of nerve growth factor (NGF) exposure and deprivation for up to 7 weeks in vitro. Most septal cultures grown 4-5 weeks with no exogenous NGF (including exposure to monoclonal or polyclonal antibodies against NGF) retained both a basal ChAT activity and the ability to increase ChAT activity in response to subsequently added NGF. When cultures were exposed to NGF (7S, 0.75 nM) for 2-3 weeks and then deprived of NGF for 2 weeks, ChAT activity fell gradually, but the number of AChE-positive neurons remained unchanged, and in many cases ChAT activity could be restored by subsequent re-exposure to NGF. Thus NGF's enhancement of ChAT activity in embryonic septal neurons in vitro is largely reversible and is not mediated by differential survival of cholinergic neurons.

Differential effects of insulin on choline acetyltransferase and glutamic acid decarboxylase activities in neuron-rich striatal cultures.

We studied the effects of insulin, nerve growth factor (NGF), and tetrodotoxin (TTX) on cellular metabolism and the activity of glutamic acid decarboxylase (GAD) and choline acetyltransferase (ChAT) in neuron-rich cultures prepared from embryonic day 15 rat striatum. Insulin (5 micrograms/ml) increased glucose utilization, protein synthesis, and GAD activity in cultures plated over a range of cell densities (2,800-8,400 cells/mm2). TTX reduced GAD activity; NGF had no effect on GAD activity. Insulin treatment reversibly reduced ChAT activity in cultures plated at densities of greater than 4,000 cells/mm2, and the extent of this reduction increased with increasing cell density. The number of acetylcholinesterase-positive neurons was not reduced by insulin, suggesting that insulin acts by down-regulating ChAT rather than by killing cholinergic neurons. Insulin-like growth factor-1 (IGF-1) reduced ChAT activity at concentrations 10-fold lower than insulin, suggesting that insulin's effect on ChAT may involve the IGF-1 receptor. NGF increased ChAT activity; TTX had no effect on ChAT activity. These results suggest that striatal cholinergic and GABAergic neurons are subject to differential trophic control.

Differential effects of acidic and basic fibroblast growth factors on spinal cord cholinergic, GABAergic, and glutamatergic neurons.

When spinal cord cultures from embryonic day 12 rats were cultured at low density, both acidic and basic fibroblast growth factors significantly increased neuronal survival and neurite outgrowth in a dose-dependent manner. The effects of acidic fibroblast growth factor were independent of heparin, in contrast to its mitogenic effects on both NIH3T3 cells and cerebral cortical astrocytes. In high-density cultures, acidic fibroblast growth factor increased choline acetyltransferase activity by 57%, glutamic acid decarboxylase activity by 58%, and aspartate aminotransferase activity by 65%. Basic fibroblast growth factor increased choline acetyltransferase activity by 73% and glutamic acid decarboxylase activity by 200% but decreased aspartate aminotransferase activity by 40%. Growing these cultures in the presence of a mitotic inhibitor did not significantly alter the effect of acidic or basic fibroblast growth factor on these enzyme activities. These results demonstrate that acidic and basic fibroblast growth factors differentially affect neurotransmitter enzyme levels of multiple classes of neurons, rather than having effects on a single neuronal population.

Protein complexity of central nervous system cell lines.

The total extracellular proteins and the most abundant 960 intracellular proteins of clonal CNS nerve and glial cell lines were examined by quantitative 2-dimensional acrylamide gel electrophoresis. While less than 0.2% of the intracellular proteins differ among the 5 nerve and 4 glial cell lines studied, over 65% of the extracellular proteins vary in distribution between the 2 major classes of CNS cells. These data indicate that the phenotypic complexity of nerve and glia populations is similar and that most of the protein complexity is in extracellular molecules.

Concanavalin A prevents acetylcholine receptor redistribution in Xenopus nerve-muscle cultures.

During neuromuscular junction formation ACh receptors accumulate at the nerve-contact region. It has been shown that this is at least partly due to lateral migration of existing receptors in the membrane (Anderson et al., 1977). Randomly diffusing ACh receptor molecules in the membrane may be trapped at the nerve-contact region to form a high receptor density area. If this were the major mechanism, cross-linking ACh receptors by tetravalent concanavalin A (Con A) should immobilize receptors and prevent nerve-induced receptor accumulation. We examined the effect of Con A on nerve-induced receptor accumulation and on the mobility of ACh receptors in cultured Xenopus muscle cells. ACh receptors were stained with tetramethyl rhodamine conjugated alpha-bungarotoxin. The cells were then treated briefly with Con A, and neural tube cells were added to these cultures. The mobility of ACh receptors was measured by the fluorescence photobleaching recovery method. The Con A treatment prevented rapid diffusion of ACh receptors as well as nerve-induced receptor accumulation. Functional synapse formation was not inhibited by this treatment. In contrast, divalent succinyl Con A did not affect the mobility of ACh receptors nor prevent nerve-induced ACh receptor accumulation. When the Con A concentration was varied, the blocking effect on the nerve-induced receptor accumulation changed in parallel with the mobile fraction of receptors. Newly inserted ACh receptors after the Con A treatment were found to be mobile and to accumulate at the nerve-contact region. In these cultures, new receptors accumulated around old, immobilized receptors in some areas along the nerve contact. This observation suggests that new receptors were inserted elsewhere and migrated to the nerve-contact region surrounding immobilized old ones. In addition to the accumulation of receptors, the nerve disperses preexisting receptor clusters prior to induction of high-density regions along the contact area, and, at this early stage, denervation disperses nerve-induced receptor clusters in Xenopus cultures (Kuromi and Kidokoro, 1984a, b). When cultures were treated with Con A, neither of these events occurred, suggesting that these are also diffusion-mediated.

In vitro glycoprotein synthesis and secretion by gastric mucosal tissue of patients with gastritis.

Endoscopically obtained gastric antrum mucosal tissue of 50 patients was examined for inflammatory cellular infiltrations and cultured. The incorporation rates of 3H-glucosamine into tissue and the secretion of 3H-labeled mucus were determined. The total glucosamine incorporated into tissue and secretions was found to be increased in mucosa from patients with histologically proven gastritis when compared to normal gastric mucosa. The differing time-dependent patterns of incorporation and secretion curves in patient groups with various forms of gastritis suggest the presence of various stimulating factors dependent on the histological pattern of inflammatory cellular infiltration. The possible modifying role of cellular and humoral factors associated with gastritis in the gastric mucus production is discussed.

[Fibrin pleurodesis in malignant pleural effusions]. / Fibrinpleurodese bei malignen Pleuraergüssen.

Fibrin pleurodesis was performed on 30 patients with malignant pleural effusions. A pleural catheter was inserted to empty the pleural space and effect the pleurodesis. When the pleural space was dry, 20 000 U of aprotinin were instilled intrapleurally for local fibrinolysis inhibition. 10 ml fibrinogen concentrate and 10 ml thrombin (500 U/ml) and 3000 KI U/ml aprotinin were then applied. Complete success was achieved in 19 of 27 patients, partial success in two. The results could not be evaluated in three patients. The only side-effect of the treatment was a subfebrile temperature in three of the 30 patients. Fibrin pleurodesis thus proved itself as an effective form of treatment with few side-effects (10%).

Formation of acetylcholine receptor clusters at neuromuscular junction in Xenopus cultures.

The formation of acetylcholine receptor (AChR) clusters at the neuromuscular junction was investigated by observing the sequential changes in AChR cluster distribution on cultured Xenopus muscle cells. AChRs were labeled with tetramethylrhodamine-conjugated alpha-bungarotoxin (TMR-alpha BT). Before innervation AChRs were distributed over the entire surface of muscle cells with occasional spots of high density (hot spots). When the nerve contacted the muscle cell, the large existing hot spots disappeared and small AChR clusters (less than 1 micron in diameter) initially emerged from the background along the area of nerve contact. They grew in size, increased in number, and fused to form larger clusters over a period of 1 or 2 days. Receptor clusters did not migrate as a whole as observed during "cap" formation in B lymphocytes. The rate of recruitment of AChRs at the nerve-muscle junction varied from less than 50 binding sites to 1000 sites/hr for alpha BT. In this study the diffusion-trap mechanism was tested for the nerve-induced receptor accumulation. The diffusion coefficient of diffusely distributed AChRs was measured using the fluorescence photobleaching recovery method and found to be 2.45 X 10(-10) cm2/sec at 22 degrees C. There was no significant difference in these values among the muscle cells cultured without nerve, the non-nerve-contacted muscle cells in nerve-muscle cultures, and the nerve-contacted muscle cells. It was found that the diffusion of receptors in the membrane is not rate-limiting for AChR accumulation.

Redistribution of acetylcholine receptors during neuromuscular junction formation in Xenopus cultures.

At the adult neuromuscular junction, acetylcholine (ACh) receptors are highly localized at the subsynaptic membrane, whereas, embryonic myotubes before innervation have receptors distributed over the entire surface. Thus sometime during development, ACh receptors accumulate to the nerve contact area. This nerve-induced receptor accumulation can be reproduced in Xenopus nerve-muscle cultures, which provides us with a unique opportunity to investigate the underlying molecular mechanism of this event. Anderson and Cohen (1977) have shown that nerve-induced receptor accumulation is, at least partly, due to migration of pre-existing receptors. It is, thus, plausible that freely diffusing receptors in the membrane are trapped at the nerve-contact region and form clusters. We tested this diffusion trap model. First, receptors in the background region are indeed predominantly mobile and those in the cluster are immobile. Second, the diffusion of receptors in the membrane is fast enough to account for the rate of receptor accumulation. Third, when receptors were immobilized by a lectin, Concanavalin A, the nerve no longer induced receptor accumulation. Thus the diffusion trap model seems adequate to accommodate these observations. Aside from this diffusion mediated mechanism, it is conceivable that newly formed receptors are preferentially inserted at the nerve contact site and these new receptors become immobilized at the site of insertion. To test this hypothesis we stained new receptors separately from old ones and quantitatively compared their distribution. For this purpose we developed a method to quantify fluorescence micrographs. We found that the ratio between old and new receptors was similar at all nerve-induced clusters examined and at the diffusely distributed region.(ABSTRACT TRUNCATED AT 250 WORDS)

[EEG changes following spirolactone treatment (author's transl)]. / EEG-Veränderungen nach Spirolacton-Behandlung.

EEG studies were done on patients with CNS disease who were treated with d-aldosterone and spirolactone. The baseline EEGs of these patients were found to be slightly abnormal. During infusion of the spirolactone derivative potassium canrenoate a transient deterioration of preexisting EEG disturbances was observed; in addition, bursts of increased muscle activity and transient relative bradycardia were seen. These findings indicate that potassium canrenoate or its lipophilic metabolite canrenone may affect the nervous system directly, possibly via a disturbance in the electrolyte balance. These results are compatible with recent findings in animal studies.

Pilot study with antilymphocyte globulin in the treatment of multiple sclerosis.

Twenty patients with multiple sclerosis, in whom treatment with azathioprine and steroids had not altered the progression of the disease, were given additional treatment with either antilymphocyte globulin (ALG) (seven patients), thoracic duct drainage (TDD) (five patients) or a combination of both (eight patients). Four of the seven patients treated with the addition of ALG showed remarkable improvement which has lasted little improvement. In the eight patients receiving both had severe allergic reactions to ALG which prevented adequate dosage. TDD alone was performed in patients sensitive to ALG. These five patients showed little improvement. In the eight patients receiving both ALG and TDD there was marked improvement in four patients which has again lasted several years. The main side effect of ALG therapy is allergic reactions. Major infections or tumour formation did not occur in any patients.