Differential binding of methyl benzimidazol-2-yl carbamate to fungal tubulin as a mechanism of resistance to this antimitotic agent in mutant strains of Aspergillus nidulans. The antimitotic compound methyl benzimidazol-2-yl carbamate (MBC) formed a complex in vitro with a protein present in mycelial extracts of fungi. The binding protein of Aspergillus nidulans showed a set of properties which is unique for tubulin. Binding occurred rapidly at 4 degrees C and was competitively inhibited by oncodazole and colchicine. Other inhibitors of microtubule function such as podophyllotoxin, vinblastine sulfate, melatonin, and griseofulvin did not interfere with binding of MBC. Electrophoretic analysis of partially purified preparations of the binding protein revealed the presence of proteins with similar mobilities as mammalian tubulin monomers. Hence it is concluded that the binding protein is identical with fungal tubulin. The effect of MBC on mycelial growth of mutant strains of A. nidulans was positively correlated with the affinity of the binding sites for this compound. The apparent binding constant for MBC and tubulin from a wild type was estimated at 4.5 X 10(5), from a resistant strain at 3.7 X 10(4), and from a strain with increased sensitivity to MBC at 1.6 X 10(6) liters/mol. Mutants showing resistance and increased sensitivity to MBC are candidates to have alterations in tubulin structure. Affinity of tubulin for MBC is probably a common mechanism of resistance to this compound in fungi. Low affinity of tubulin for MBC is probably a common mechanism of resistance binding constant of 2.5 X 10(3) liters/mol.


[Ultra-selective vagotomy in the treatment of duodenal ulcer. Report on the round table discussion (77th French Surgery Congress, 1975)]. This panel enables us to draw following conclusions on highly selective vagotomy for duodenal ulcer: --One fact seems to be sure: H.S.V. is a benign and efficient operation over short and average periods of time, providing that some technical requirements are observed. --However some points are still under discussion and much debated: --the useful purpose of per-operative tests; --the more or less frequent addition of drainage procedure. --Lastly, long-term anti-ulcerous efficiency (over 5 years) is still not known precisely; some think that the risk of vagal regeneration are greater after this type of vagotomy.


Chromatographic characterization of phenothiazine drugs by a reversed-phase thin-layer technique. A reversed-phase thin-layer chromatographic technique was used for the characterization of 26 phenothiazine drugs. With two chromatographic systems having the sam stationary phase and phase volume ratio, but mobile phases of different pH, all but two of the compounds could be identified. Rf values in the different systems were standardized by applying a reference compound to the plates next to each compound under investigation; the corrected Rf values were calculated from the differences in the Rm values of the compounds and the reference compound, and the theoretical Rm value of the reference. It was shown that Rf values for different chromatographic systems with the same stationary phase could be predicted with reasonable accuracy. The pH of the mobile phase, for which a maximum difference in Rf values was obtained for pairs of compounds, could also be calculated and corresponded well with the observed values.


Use of acetonitrile for the extraction of herbicide residues from soils. Using a 10% aqueous acetonitrile solution for extraction and an identical solvent clean-up procedure, soil-based residues of the herbicides alachlor, benzoyl-prop-ethyl, flufenprop-isopropyl, flufenprop-methyl, dichlorfop-methyl, nitrofen, and profluralin were recovered reproducibly from three prairie soils fortified at 0.5 and 0.1 ppm levels. The acidic herbicides benazolin, 2,4-D, and 2,4,5-T, together with the acids derived from benzoylprop-ethyl, dichlorfop-methyl, flufenprop-isopropyl, and flufenprop-methyl were reproducibly recovered from the three prairie soils fortified at 0.5 and 0.1 ppm levels using 30% aqueous acetonitrile containing 1% acetic acid after identical clean-up stages. All compounds were analysed by gas chromatographic means utilising an electron-capture detector. The two procedures described were developed for the routine extraction and analysis of neutral and acidic herbicide residues from field soil persistence studies.


High-pressure liquid chromatography of cannabis. Quantitative analysis of acidic and neutral cannabinoids. A reversed-phase high-pressure liquid chromatographic method has been developed for the simultaneous analysis of the acidic and neutral cannabinoids in cannabis. Cannabigerol and cannabigerolic acid have been located in the liquid chromatogram of cannabis and factors affecting the chromatographic process are discussed. A method for quantitating one component in the presence of a second unresolved component is described.


Residual anionic properties of a covalently substituted controlled-pore glass, glyceryl-CPG. A commercial controlled-pore glass medium for exclusion chromatography which is substituted with glycerol to eliminate non-specific adsorption (Glyceryl-CPG) was examined. Experiments on the elution of acetylcholinesterase and ganglioside micelles at varied ionic strength and pH showed that a slight anionic character still persisted on the glass matrix. At ionic strengths above 0.1, this had no effect on the elution of proteins. The material was found to have no tendency to adsorb proteins and other compounds, and was judged an excellent medium for exclusion chromatography. As a support for affinity chromatography, Glyceryl-CPG could be activated by periodate to form a virtually neutral matrix-ligand system.


Inhibition of human plasma renin activity by pepstatin. Pepstatin, a pentapeptide isolated from streptomyces, is a powerful inhibitor of several acid proteases. Its ability to inhibit the renin-angiotensinogen reaction was studied in vitro, in various human plasma. A 50% inhibition of plasma renin activity was obtained with 10(-6)M pepstatin at pH 5.7 and 10(-5)M at pH 7.4. The inhibition of plasma renin activity by pepstatin was studied in hypertensive patients with various plasma renin levels. The inhibitory effect could be demonstrated in patients with low, normal and high renin activity at both pH's. The type of inhibition and the inhibitory constant were investigated by Dixon plot and Lineweaver-Burk plot in three separate experiments. On both representations, a competitive type of inhibition was found with an inhibitory constant of about 1.2 x 10(-6)M.


Adaptive responses of brain cyclic AMP-generating systems to alterations in synaptic input. The concept of subsensitivity and supersensitivity as a mechanism of neuronal adaptation to alterations in synaptic activity in the brain is an attractive one. However, the complexity of the central nervous system has made it difficult to determine the cellular basis of apparent changes in neuronal excitability resulting from alterations in synaptic input (cf. ref. 83). It now seems that, at least for inhibitory central pathways in which cyclic AMP-generating systems mediate postsynaptic receptor-responses, alterations in synaptic input lead in rat cortex to predictable super- or subsensitivity of the norepinephrine-sensitive cyclic AMP-systems. Supersensitivity of histamine-sensitive cyclic AMP-systems also occurs in rat cortex as a result of apparent lesions of histaminergic tracts. The reason that supersensitivity does not develop to norepinephrine and histamine in guinea pig cortex after similar "denervations" is not known, but is central to an understanding of the factors involved in the role of cyclic AMP-mechanisms to the adaptive plasticity of the central nervous system. However, even before an understanding of all the factors involved in alterations in cyclic AMP-responses in brain tissue is obtained, studies on the effects of environmental and drug-manipulations can provide insights both into the central roles for cyclic AMP-mechanisms and into the nature of drug action. Clearly, an understanding of the interelationships of synaptic input and adaptive changes in postsynaptic cyclic AMP-systems and the exploitation of such knowledge for the elucidation of central adaptive function is an exciting challenge for future research.


Enhancement of methacholine-stimulated guanosine 3':5'-cyclic monophosphate formation in supersensitive guinea pig vasa deferentia. Decentralization of the guinea pig vas deferens resulted in supersensitivity of the contractile response to methacholine. Methacholine also elevated cyclic GMP to a greater extent in the decentralized vas deferens than in sham-operated controls. Decentralization did not alter basal cyclic AMP or cyclic GMP content. These observations are analogous to the enhanced responsiveness of cyclic AMP formation in supersensitive beta adrenergic systems.


Inhibition of mammalian soluble guanylate cyclase activity by adenosine 5'-tetraphosphate, guanosine 5'-tetraphosphate and other nucleotides. The effects of a variety of purine and pyrimidine nucleotides were tested for their capacity to inhibit mammalian soluble guanylate cyclase activity. Adenosine 5'-tetraphosphate (ATetP), ATP, ADP, AMP, guanosine 5'-tetraphosphate (GTetP) and GDP were found to inhibit soluble guanylate cyclase activity from rat lung and other mammalian tissues. The corresponding cytosine and thymine nucleotides showed little or no inhibitory activity, except for thymidine 5'-tetraphosphate, which inhibited glanylate cyclase activity but to a lesser extent than did the purine nucleoside tetraphosphates. ATetP and GTetP were found to be potent inhibitors of soluble guanylate cyclase activity from rat, guinea pig and mouse lung, rat heart and rat brain. Both purine nucleoside tetraphosphates were competitive inhibitors of the rat lung soluble enzyme. ATetP and GTetP had Ki values of 1 muM and 2.5 muM, respectively. The experimental data suggest that purine nucleoside tetraphosphates, and perhaps other purine nucleotides, may play a biologic role in modulating mammalian soluble guanylate cyclase activity.


Enhancement of the antiplaque value of antibacterial agents through enamel-conditioning methods: II. Acquisition of antiplaque properties by treated enamel. Enamel specimens treated with systems containing enamel conditioners and antibacterial agents have previously been shown to incorporate the latter into the enamel. It has now been demonstrated that enamel blocks treated with these systems become highly resistant to bacterial colonization, that this effect is rather long lasting, and that the treated specimens prevent acid formation when incubated with Streptococcus mutans in a sugar-containing medium.


The systemic use of procaine in the treatment of the elderly: a review. This article is a review and evaluation of the world literature on the systemic use of procaine in the treatment of the aging process and the common chronic diseases of later life. Included are data from 285 articles and books, describing treatment in more than 100,000 patients in the past 25 years. Except for a possible antidepressant effect, there is no convincing evidence that procaine (or Gerovital, of which procaine is the major component) has any value in the treatment of disease in older patients. If procaine has an antidepressant effect, there is some likelihood that this accounts for the reports of decreased complaints referable to the musculoskeletal, cardiovascular, endocrine sexual, gastrointestinal and respiratory systems.


[Diagnosis of fetal distress during labor with the aid of the Hammacher cardio-tocographic score and fetal pH determination]. The authors study the value of the Hammacher tocographic scoring system in the diagnosis of fetal distress in labour. This score gives equal value to three components of the fetal heart rate: the base line, transient variations (dips) and fluctuations (oscillations). There is a statistically very significant (p less than 0.001) relationship between the cardio-tocographic score and the fetal pH as has been found in 106 labours with a coefficient of correlation of 0.67. The score can only be carried out usefully when a "beat to beat" to heart rate is registered and when this is not influenced by the administration of drugs to the mother. The score has been demonstrated to be particularly useful in the diagnosis of fetal distress when the membranes are still intact.


[The risk of cardiac arrest during laparoscopy. A study of 50,000 laparoscopies and animal experimentation]. In a French national enquiry into 50,000 laparoscopies it emerges that the number of cardiac arrests works out at 1 in 2,000. The percentage of reversible cardiac arrests was 1 in 2500 and of irreversible cardiac arrests 1.2 in 10,000. 25 cases were studied in great detail, both clinically and physiopathologically. This was thanks to a very full questionnaire most kindly filled up by those who had told us that this type of accident had happened in their practice. The physiopathogenesis of circulatory arrests during laparoscopy is complex. It appears however that among the different potentiating factors are changes in haemodynamics and biological changes that give rise to the greatest risks. Experimental work carried out with a long-tailed baboon was performed by the authors with the idea on the one hand of confirming that haemodynamic and biological troubles resulting from the pneumoperitoneum were important and on the other hand of analysing the changes in detail and of working out prophylactic principles for the anaesthetist and the laparoscopist.


Human epidermal transglutaminase. II. Immunologic properties. A monospecific antibody for human epidermal transglutaminase was prepared in rabbits. The antibody formed single immunoprecipitin lines with purified or crude human transglutaminases and quantitatively precipitated transglutaminase activity. There were no precipitin reactions between human factor XIII (zymogen or active enzyme) and antihuman epidermal transglutaminase or between human epidermal transglutaminase and antihuman factor XIII. Heating epidermal transglutaminase (56 degrees C, 15 min) in the presence of calcium increased the enzyme activity up to 10 times baseline levels. The heat-activated human epidermal transglutaminase was identical by immunodiffusion with the native enzyme, although slightly higher precipitation titers were detected following heating. There was no cross-reaction of antihuman epidermal transglutaminase with frog, rat, mouse, chicken, or human hair follicle transglutaminases.


Interactions between viruses and bacteria in patients with chronic bronchitis. The possibility that viral infections of the respiratory tract might predispose to bacterial colonization or infection was studied in 120 patients with chronic obstructive pulmonary disease and 30 control subjects; these individuals were observed for seven years. The ratio of the number of observed to the number of expected associations between viruses and bacteria was 2.43 (P = 0.037) for the pair influenza virus and Streptococcus pneumoniae and was 2.06 (P = 0.056) for influenza virus and Haemophilus influenzae. Consistently positive, but not significant, associations were detected between rhinovirus and herpes simplex virus infections and isolations of S. pneumoniae and H. influenzae. In contrast, isolations of the nonpathogenic Haemophilus parainfluenzae could not be related to prior viral infections. Significant rises in titer of antibody to H. influenzae were detected on 76 occasions, and 20 (26%) of these antibody rises were associated with viral or mycoplasmal infections during the preceding 120 days. The expected number of such associations was 8.34 (ratio of number observed to number expected, 2.40; P = 0.08). These results suggest that viral infections of the respiratory tract in patients with chronic obstructive pulmonary disease are associated with increased colonization by potentially pathogenic bacteria and may also predispose to infections with H. influenzae.


Experimental otitis media due to Streptococcus pneumoniae: immunopathogenic response in the chinchilla. Acute otitis media was produced in 110 chinchillas by inoculation of type 23 Streptococcus pneumoniae directly into the middle ear cavity by tympanotomy. During the first three days after inoculation, inflammatory cells were seen in the mucoperiosteum of the middle ear. After four to seven days, there was purulent exudation in the middle ear cavity, and 40% of the animals had pneumococcal meningitis and/or bacteremia. The middle ears were sterile in five of 28 animals sacrificed during the second week and in six of seven animals sacrificed at six weeks, although subepithelial changes persisted in the mucoperiosteum. Levels of antibody to S. pneumoniae in serum were measured by radioimmunoassay; mean values were 6.1 ng of pneumococcal antibody nitrogen/ml in 28 uninfected control animals and 16.5 ng of antibody nitrogen/ml in 29 animals sacrificed two weeks after inoculation (P less than 0.025). Opsonic activity of serum against S. pneumoniae was evaluated in infected and uninfected chinchillas. The opsonic titer was significantly higher in infected animals sacrificed at six weeks than in uninfected controls. Although pneumococcal polysaccharide antigen was found by counterimmunoelectrophoresis in 25 of 30 middle ear effusions, it could not be detected in the serum from infected animals. Methods for infection and sacrifice of chinchillas yielded reproducible results. This model should permit evaluation of the pathologic response to other serotypes of S. pneumoniae and possibly to prophylactic and therapeutic regimes.


Inhibition of vitamin B12 binding to transcobalamin II at low pH: basis of a procedure for quantitation of circulating TC II and R binders. A diminution in the binding of exogenous vitamin B12 by serum or plasma at pH 1.5 to 2 (acid-resistant binding capacity, ARBC) as compared with the binding capacity at neutral pH (unsaturated vitamin B12 binding capacity, UBBC) was observed. This phenomenon was found to be attributable to the absence of transcobalamin II (TC II)-associated vitamin B12 from serum labeled at low pH, as demonstrated by gel chromatography on Sephadex G-200. Further confirmation was obtained by demonstration of a significant correlation between the ARBC and the R binder content, quantitated as resistance to precipitation by ammonium sulfate at a 2M concentration. Serum labeled at acid pH failed to deliver vitamin B12 to Hela cells indicating absence of a "functional" TC II-B12 complex. The differing vitamin B12 binding capacities of neutral and acidified material was utilized to fractionate the unsaturated vitamin B12-binding proteins of serum and plasma. The ARBC was used to measure the R binder content, and TC II was calculated from the difference between UBBC and ARBC. The fractionation procedure was performed on 75 sera and showed increased ARBC in patients with myeloproliferative disorders and decreased ARBC in leukopenia. The content of unsaturated vitamin B12-binding protein was compared in 75 paired samples of serum and plasma collected from EDTA-anticoagulated blood containing sodium fluoride to inhibit release of granulocyte binders. The ARBC content of fluoridated plasma was significantly lower, due to decreased in vitro R binder release. Plasma also contained less TC II, possibly related to in vitro cellular uptake of this binder in fluoridated plasma.


Properties and development of erythropoietic stem cells in the chick embryo. 1. When injected into irradiated chickens, haemopoietic stem cells give rise to well-defined erythrocytic colonies in the host marrow. Such stem cells (CFU-M = Colony Forming Unit in Marrow) have been found in different tissue of the chicke embryo (yolk sac, blood, marrow). Analysis of the properties of CFU-M reveals that they represent two classes of stem cells: pluripotent stem cells mainly in adult marrow and erythrocytic-committed stem cells present in yolk sac. 2. Yolk sac contains the main pool of CFU-M during the major part of embryonic life. In the blood of 6-day-old embryo, there are three or four times more CFU-Ms than in the yolk sac; they are no longer detected in the blood after the 16th day of incubation. During development of the marrow, stem cells are actively differentiating and their total number remains the same from 16 days to hatching.


Temperature acclimation and oxygen-binding properties of blood and multiple haemoglobins of rainbow trout. Acclimation of rainbow trout to 5, 15 and 22 degrees C for periods exceeding 4 months had no significant effect on the oxygen affinity of whole blood or on the concentration of ATP, which is the main organic phosphate in red cells. Slight differences were, however, found in the oxygenation properties of the haemolysates, which correlate with changes in the relative concentration of the multiple haemoglobins. The oxygen-binding properties of the main haemoglobin components account for the observed differences in the haemolysates. The possible thermoacclimatory significance of changes in haemoglobin multiplicity and co-factor concentrations is discussed.


An electrophysiological study of mechanisms controlling polyp retraction in colonies of the scleractinian coral Goniopora lobata. 1. Electrical or mechanical stimulation of Goniopora lobata produces coordinated retraction of polyps in the colony. With repetitive stimulation, the response spreads in linear, radial increments which become successively smaller with each stimulus. 2. Electrical activity recorded from these colonies is interpreted as originating in a conduction system responsible for effecting the colonial retraction response. The electrical activity spreads incrementally through the colony in a similar manner to the behavioural response. 3. Various hypotheses have been proposed to account for such a spread of electrical acitvity. Of these, only interneural facilitation is of appreciable importance to Goniopora. 4. Temporary termination of a pathway, by the passage of an impulse through it, was found and interpreted as being an additional and important property of the colonial conduction system.


Possible involvement of monoamines in the release of adipokinetic hormone in the locust Schistocerca gregaria. 1. The adipokinetic hormone release, which can be induced by anticholinesterases, is reduced by depleting the content of monoamines in the nervous system. 2. The participation of monoamines in the pathway of release of adipokinetic hormone is studied in vivo and in vitro. 3. A possible mechanism for anticholinesterase-induced release of this hormone involving cholinergic and aminergic transmission is postulated.


Oxygen dissociation curves of the blood of larval and adult lampreys (Lampetra fluviatilis). 1. An electrolytic method was used to plot the oxygen dissociation curves of whole blood from both the larva and adult of the lamprey Lampetra fluviatilis at a temperature of 10 degrees C and over a pH range of 6-5-8-1. 2. Larval blood has a far higher affinity for oxygen than that of adults, the respective calculated P50's at a pH of 7-75 being 1-9 and 10-7 mmHg. 3. The high affinity of larval blood is of use to a relatively sedentary animal living in burrows, and the increased oxygen delivery pressure brought about by the shift of the curve to the right in the adult is of advantage to an animal exhibiting greater activity. 4. The n value obtained from the Hill plots increased with increasing saturation and were lower in larvae than adults at the same level of blood saturation. 5. The Bohr effect in larvae at 10 degrees C over the pH range 6-5-8-1 was --0-25, a value which did not differ significantly from the -0-22 found in adults.


Specific transplantation tolerance induced by autoimmunization against the individual's own, naturally occurring idiotypic, antigen-binding receptors. Serum or urine from normal adult Lewis rats can be shown to contain detectable amounts of idiotypic, antigen-binding receptors with specificity for the major histocompatibility complex locus antigens of the rat, the Ag-B locus antigens. Such purified naturally occurring receptor molecules, be they of T- or B-lymphocyte origin, can be used in a polymerized form to provoke the production of auto-anti-idiotypic antibodies when injected back into normal Lewis rats. As a consequence of this autoimmunity, lymphocytes of these Lewis rats can be shown to be depleted of cells carrying the relevant idiotypic receptors signifying reactivity against a given Ag-B locus-determined antigen(s). This specific lack of idiotypic lymphocytes is manifested as a selective loss of reactivity against the relevant Ag-B-incompatible antigens as measured by graft versus host or MLC reactions. Furthermore, autoimmune Lewis rats display specific transplantation tolerance against the skin grafts from the relevant strain, as demonstrated by specific prolongation of graft survival. A further indication of the specific tolerence state of these rats comes from the highly reduced ability to produce circulating antibodies against the relevant Ag-B antigens. No side effects of these autoimmunization procedures have been noted so far. It would thus seem clear that a prolonged state of specific transplantation tolerance can be achieved via autoimmunization against the individual's naturally occurring idiotypic, antigen-binding receptors.


Respiratory behaviour of sea-urchin spermatozoa. I. Effect of pH and egg water on the respiratory rate. Effects of pH and egg water on the respiration of sea-urchin spermatozoa were polarographically studied in three sea-urchins and one starfish species. Sea-urchin sperm respiration is extremely sensitive to change in the pH of the suspending medium over a wide range. In normal-sea water, the pH of the sperm suspension decreased from 8.02 to 7.62, after four to five minutes' incubation at 18 degrees C. The Respiratory Dilution Effect could be recognized in the same medium. However, when sea water was buffered with HEPES at pH 8.2, the Effect was no longer observed. The diffusate from egg water (jelly coat solution) brought about a striking increase in the respiration when added to moderately respiring spermatozoa in HEPES-sea water of pH values lower than 7.9. No inccrease in the respiration was observed when the diffusate was added to vigorously respiring spermatozoa in HEPES-sea water of pH values higher than 8.2. Sperm motility was also inhibited by acid pH, and this inhibition was reversed by the addition of the diffusate. It does not seem that there is any species-specificity among three sea-urchins and one starfish used. The role of the diffusate is discussed in relation to the penetration of spermatozoa through the jelly coat to the egg surface.


Respiratory behaviour of sea-urchin spermatozoa. II. Sperm-activating substance obtained from jelly coat of sea-urchin eggs. A sperm-activating substance (SAS) was obtained from the jelly coat of sea-urchin ova and its chemical properties were investigated in three sea-urchin species. The SAS was partially purified from the jelly coat of Pseudocentrotus eggs through several steps of purification by procedures consisting of charcoal adsorption, ion-exchange chromatography on DEAE-Sephadex A-25 column, and gel-filtration on Sephadex G-15 columns. The partially purified SAS was found to contain a ninhydrin-positive material and is inactivated by pronase digestion. The molecular weight of SAS was estimated as about 630 by gel-filtration through Sephadex G-25 and the isoelectric-point of SAS is located at about pH 5.3 by isoelectrofocusing method. The SAS is non-volatile, alcohol-soluble, and labile in a diluted alkaline or acid solution. The origin of SAS is discussed.


Deficiency of NADPH oxidase activity in chronic granulomatous disease. NADPH oxidase activity was examined in paired 27,000 x g granule fractions isolated from normal polymorphonuclear leukocytes from patients with chronic granulomatous disease. At 0.17 mM NADPH, the oxidase activity was not measurable in normal resting cells but was activated by phagocytosis. This activation was absent in CGD cells. At higher levels of NADPH, activity was present in cells from patients with CGD, although it was lower than normal, and no difference in activity was found between resting and phagocytizing cells. Granule fractions from phagocytizing normal cells exhibited higher than granule fractions from resting normal cells at all levels of NADPH. These results suggest that NADPH oxidase activity is defective in chronic granulomatous disease, and further that the defect is not the absence of the enzyme but rather a failure to activate it.


Development of the Timor filaria in Aedes togoi: preliminary observations. Developmental stages of the Timor filaria recovered from experimentally infected Aedes togoi mosquitoes are described. Mosquitoes were dissected and examined for larvae beginning 1 1/2 days and continuing daily for 9 days after they had fed on a carrier on the island of Flores, Indonesia. Timor microfilariae develop rapidly to third-stage larvae within the thoracic muscles of A. togoi: the first molt occurs at 3 1/2 days, the second molt as early as 5 1/2 days, and infective forms are found at 6 1/2 to 7 1/2 days postfeeding. These larvae were compared with similar stages of subperiodic Brugia malayi recovered from A. togoi fed on an infected jird (Meriones unguiculatus) and the features distinguishing larvae of the Timor filaria from those of B. malayi are restricted to the tails of the first-stage forms. The terminal and subterminal nuclei of the Timor larva are generally smaller than those of B. malayi, and there is little if any bulge of the cuticle around them. A constriction of the tail between these nuclei is subtle or absent. The findings of this study support the view that the Timor filaria is a member of the Brugia complex.


Acetylcholine synthesis in sympathetic human neuroblastoma. The synthesis of acetylcholine, as well as catecholamines, was studied by assaying the activities of choline acetyltransferase (ChA) and tyrosine hydroxylase (TH) in the tumor tissues and the culture cells of human neuroblastoma. In the majority of 20 neuroblastomas of sympathetic origin, both ChA and TH activities were detected at a significantly high level. In the culture cells of five cell lines of human neuroblastoma, ChA activity was high, but TH was negative in four of the lines. However, it was observed that these enzyme activities changed significantly while in the long-term culture. ChA assay is a useful diagnostic test for neuroblastomas that synthesize acetylcholine. Future studies of neuroblastoma should consider cholinergic activity.


Some biological and pharmacological properties of inflammatory exudates. Inflammatory exudates were obtained from polyester sponges which had been implanted subcutaneously in rats four days previously. This material was found to be anti-inflammatory when injected into other rats in which carrageenan pleurisy had been induced. At a dose of 600 mg kg-1 exudate inhibited the formation of pleural effusion, emigration of both neutrophils and mononuclear cells and the accumulation of beta-glucuronidase and lactic dehydrogenase. The same dose of sponge exudate did not however inhibit the increased vascular permeability induced in the rat skin or rat foot following injection of 5-hydroxytryptamine, histamine, prostaglandin E1, or bradykinin. Furthermore sponge exudate did not reduce the haemolytic complement titre of rat serum either in vivo or in vitro. The possible mechanism of anti-inflammatory action of exudate is discussed.


The role of copper in preventing gastrointestinal damage by acidic anti-inflammatory drugs. The ability of several non-steroidal acidic anti-inflammatory drugs to cause ulceration when given as copper complexes has been examined. The damage caused by clopirac, niflumic acid and aspirin was virtually abolished when they were given as copper complexes whereas the damage caused by indomethacin, ketoprofen and (+)-naproxen was unaltered. The lack of ulceration with three of these preparations appeared to be correlated with a much reduced ability to inhibit prostaglandin synthesis as determined using an in vitro enzyme system.


Methods for study of fluphenazine kinetics in man. Fluphenazine and its principal metabolites, fluphenazine sulphoxide, and 7-hydroxyfluphenazine were identified and quantified in human plasma, urine and faeces following intramuscular and oral administration of 14C-fluphenazine dihydrochloride. The presence of a conjugate fraction was also noted. Unmetabolized fluphenazine was selectively extracted into n-heptane. The metabolites were separated by solvent extraction into toluene. Conjugates were hydrolysed back to fluphenazine, fluphenazne sulphoxide and 7-hydroxyfluphenazine. Fluphenazine and fluphenazine conjugates were also measured in the urine of patients receiving long term non-radioactive fluphenazine decanoate therapy. The urinary excretion rate of the conjugate fraction was systematically related to the plasma concentration, regardless of urine flow rate or pH, providing a convenient method for the assessment of fluphenazine kinetics by urinary excretion studies not involving administration of labelled drug.


Potentiation of dopaminergic transmission by phosphodiesterase inhibitors and cyclic nucleotides. Experiments were made to determine whether cyclic AMP plays a role in transmission at identified dopaminergic synapses in the water snail Planorbis corneus. Intracellular stimulation of a specific dopamine neuron produces direct inhibitory postsynaptic potentials (ipsps) in a number of other neurons. These ipsps, which are mediated by dopamine, were potentiated by as much as 120% by caffeine, theophylline or dibutyryl cyclic AMP, although they were unaffected by cyclic AMP and prostaglandin E1. Caffeine and theophylline also potentiated the inhibitory response to dopamine, applied to the postsynaptic neurons by perfusion or iontophoresis, but the effects were generally much smaller (maximum potentiation 30%). The results provide evidence that postsynaptic cyclic AMP is involved in transmission at these synapses, but that the phosphodiesterase inhibitors may also have a presynaptic effect.


A simple technique for the frequent intravenous infusion of fluid without heparin into the rabbit. A technique is described for the daily intravenous injection over 16 weeks in the rabbit. The system consists of a Teflon cannula passed down the external jugular vein with the tip in the superior vena cava. The cannula is joined at the point of entry to the jugular vein to a length of silicon rubber tubing, which is then passed subcutaneously to the forehead. The silicon tubing is terminated on a Luer needle hub, which is held in a simple Perspex plate secured subcutaneously between the rabbit's ears. The Luer hub is covered with a replaceable rubber cap through which injections may be made. With this system cannulae were maintained patent for 16 weeks by flushing once a day with Hanks' balanced salt solution, pH 7-4.


Anabolic and androgenic activities, in rat, of some nandrolone and androstanolone esters. The anabolic and androgenic activities of the formate to undecanoate esters of nandrolone and formate to valerate esters of androstanolone, after intramuscular injection, have been determined in rat. The response to a given dose was measured as cumulative weight (the area under the plot of weight of indicator organ against time). Levator anus muscle was used to assess anabolic activity, and the sum of the cumulative weight for prostate and seminal vesicles for androgenic activity. Log dose-log cumulative weights plots were parallel, and biological activities were expressed as the cumulative weight corresponding to a 2 muM dose, calculated from the regression lines. Anabolic-androgenic ratios for both series were calculated and were found to be minimum in the region of the propionate and butyrate. The anabolic-androgenic ratios of the nandrolone esters continued to increase after the minimum, as the series ascended. This method is believed to give a reliable assessment of anabolic and androgenic activities of steroid esters.


The adhesion of film coatings to tablet surfaces--instrumentation and preliminary evaluation. The strength of the adhesive bond between a film coating and a tablet surface has been studied using a specially designed tensile testing apparatus. The adhesion has been taken as the force required to remove the film from unit area of the tablet surface and has been shown to be dependent on the compression pressure used to prepare the tablet and on changes in the film formulation. Although plasticizers did not show any significant effect on the adhesion of a hydroxypropyl methylcellulose film, a reduction of 45% was found on the addition of 10% w/w titanium dioxide.


Size analysis of metered suspension pressurized aerosols with the Quantimet 720. A method is described for particle sizing of pressurized metered suspension aerosols by collection in a settling drum followed by microscopic evaluation of the slides with a Quantimet 720 automatic image analyser. The method gives satisfactory representation of the distribution of particles settling to the drum base, and demonstrates the excellent stability and reproducibility between and within aerosol packs for two widely used inhalation products. There is much drug deposition of somewhat finer size distribution on the wall of the drum than on the drum base. In spite of this wall loss, the method gives only slightly higher results for the weight and number mean diameters, than when both wall and base distributions are considered. The Quantimet was found to be suitable for particle sizing salbutamol used in preparing aerosol products.


Size analysis of suspension inhalation aerosols by inertial separation methods. The particle size distribution of beclomethasone dipropionate (BDP) aerosols delivered from pressurized metered dose suspension inhalers has been measured with three cascaded inertial separation instruments, the Casella Cascade Impactor, Multistage Liquid Impinger and Cascade Centripeter. Various methods for collecting the emitted aerosol before measurement have been examined. A bent glass tubular 'throat', used as a simulated oro-pharynx, collects 35-60% of the emitted dose by impingement of the wet spray cone in the throat. The aerosol passing through the throat has a similar but somewhat finer size distribution to that collected by firing directly into a large flask. The three cascaded instruments give similar results which in the Multistage Liquid Impinger also resemble those given by a salbutamol inhaler. The mass fraction (35-60%) emitted from the oral adaptor which is of a size capable of deep lung penetration ( less than 4 mum) is much higher than the fraction (10-16%) found in the lungs of dogs after inhalation of aerosol. The size distributions resemble those determined by microscopy and are expressed as aerodynamic sizes, thus showing that the particles approximate to unit density spheres. The performance of two simpler devices, Kirk's apparatus and the Harwell size selective air sampler are also assessed, the latter shows some promise for the simple evaluation of the respirable fraction of inhalation aerosols.


An investigation into the deposition of inhalation aerosol particles as a function of air flow rate in a modified 'Kirk Lung'. The effect of air flow rate on the deposition of inhalation aerosol particles in a modified 'Kirk Lung' has been studied. Two commercial products were used. The amount depositied in the mouthpiece and throat regions decreases with increasing flow rate. The amount in the collector rises to maximum of approximately 50%. This material represents the therapeutically available portion. Such an apparatus would be useful for routine quality control of the size distribution ratios for inhalation aerosol particles.


The in vitro dissolution of phenobarbitone sodium from ethyl cellulose microcapsules. Microcapsules of sodium phenobarbitone, with a wall of ethyl cellulose, have been prepared. The size distribution was determined by use of standard sieves and the effect of coreWALL ratio noted. Release of the drug into an aqueous medium was studied. The release pattern was found to have similar characteristics to the release of a drug from an insoluble porous matrix.


The granulation of binary mixtures: the effects of the properties of the component powders on granules. Sulphanilamide and citric acid individually and in various proportions with lactose, have been granulated by massing and screening. There was an optimum blend, that produced granules of maximum mean size and strength, for each binary system examined. The proportion of the components of this optimal blend was dependent on the physical properties of the second component in a mixture with lactose. Results from three systems, lactose:boric acid, lactose:sulphanilamide and lactose:citric acid indicate that although part dissolution of powder during granulation is a factor affecting granule properties, in some systems other physical properties of the second component may become dominant. It is suggested that the combined effect of cohesiveness and wettability of the powders may make the major contribution to granule strength with the sulphanilamide systems. The ultimate mean granule size produced is determined by the wettability or solubiluty of the powders, or both, in all cases examined. The great affinity of citric acid for aqueous binder solution was the dominant factor determining the properties of granules prepared from lactose:citric acid mixtures.


Stereochemical studies of adrenergic drugs. Optically active derivatives of imidazolines. The synthesis of (R)-(+)-4-methyl-2-(1-naphthylmethyl)imidazoline hydrochloride (2) and (S)-(-)-4-methyl-2-(1-naphthylmethyl)imidazoline hydrochloride (3) is presented. The synthesis involves the preparation of (R)-(+)- and (S)-(-)-1,2-diaminopropane dihydrochloride and then allowing the appropriate diaminopropane to react with ethyl 1-naphthyliminoacetate hydrochloride in the presence of triethylamine. The parent compound, naphazoline, is a potent alpha-adrenoreceptor agonist (-log ED50 = 7.22), whereas the methylated derivatives, 2 and 3, were moderately potent antagonists (pA2 = 5.6 and 5.8, respectively) of the alpha-adrenoreceptor. Compounds 2 and 3 also produced blockade of the response to histamine on the rabbit aorta, but at concentrations approximately 20 times higher than necessary to produce equal blockade of the alpha-adrenoreceptor.


Effects of staphylococcal products on locomotion and chemotaxis of human blood neutrophils and monocytes. The effects of staphylococcal products as chemo-attractants for human blood neutrophils and monocytes and as inhibitors of locomotion of these cells were studied with bacterial cells, culture filtrates and isoelectrically focused fractions from culture filtrates of nine strains of Staphylococcus aureus. Little direct chemotactic activity of staphylococcal products for neutrophils was observed, although a chloroform-soluble extract of the whole organisms contained such activity. The major chemotactic effect of staphylococci for neutrophils was indirect, i.e., generated when the organisms or their products were incubated with plasma, perhaps due to activation of complement. In contrast, direct chemotactic activity for monocytes was found in a large number of staphylococcal fractions. Staphylococci also produced inhibitors of locomotion of both neutrophils and monocytes. Isoelectric focusing showed more fractions inhibitory for neutrophils than for monocytes. Some of the inhibitors could be identified. Staphylococcal alpha-toxin inhibited migration of both neutrophils and monocytes. Sphingomyelinase C (beta toxin) inhibited migration of monocytes but not of neutrophils. Leucocidin-rich strains were strongly active as inhibitors of neutrophil locomotion but less so as inhibitors of monocyte locomotion.


Osteomyelitis following puncture wounds of the foot in children. Review of the laboratory and clinical findings and treatment of eight patients with osteomyelitis of the foot after puncture wounds revealed that: 1) osteomyelitis after puncture wounds is a infrequent but potentially serious complication, with significant morbidity; 2) osteomyelitis is frequently preceded by inadequate primary care for simple puncture wounds, and when treatment is appropriate, osteomyelitis usually can be avoided; 3) P. aeruginosa is the most commonly recovered organism; 4) the clinical presentation is characterized by a lack of systemic toxicity, paucity of laboratory abnormalities, and evidence of a localized infection process and the patient may be asymptomatic for a few days to several months after the injury before presentation of the osteomyelitis; and 5) once the infection has become established, treatment must be aggressive, including surgical debridement.


Ureteral changes in polyarteritis nodosa as seen during excretory urography. The conventional excretory urogram may provide valuable information in the diagnosis of polyarteritis nodosa. A finding of imprints on the ureteral wall or a nodular appearance of the ureter simulating a string of pearls would seem to be pathognomonic for this disorder. Two cases with these manifestations are presented and the differential diagnosis is discussed.


The incidence of intersexuality in patients with hypospadias and cryptorchidism. The incidence of intersexuality was determined in 45 patients with cryptorchidism and hypospadias seen between 1952 and 1975. Including patients with ambiguous genitalia the incidence was 53 per cent. When patients with ambiguous genitalia were excluded, providing a group more representative of the patient population usually seen by the urologist, a diagnosis of intersexuality was made in 27 per cent. The concomitant occurrence of hypospadias and cryptorchidism should alert the urologist to evaluate the patient for the presence of intersexuality.


Calcium oxalate crystalluria.

Calcium oxalate crystalluria. Calcium oxalate crystals were obtained from urine specimens submitted to a hospital laboratory. The incidence of crystalluria was 4.2 per cent of 42 times the maximum reported incidence of urinary calculi. In our opinion the crystalluria was real and not artifactual. The crystalline structure was determined by polarized light, x-ray diffraction and electron microprobe analysis. Calcium oxalate occurs in several forms--the dihydrate as bipyramidal and dodecahedral prisms, the monohydrate as biconcave ovals, dumbbell shapes and intermediate forms.


Congenital megalourethra associated with prune belly syndrome. Dilation of the penile urethra is described in a male infant with the classic triad of the prune belly syndrome. This dilated penile urethra was not accompanied by obstruction of the urethra proximal or distal to the dilation. Dilation of the penile urethra is a rare feature of the prune belly syndrome and may be part of the same developmental abnormality that affects the remainder of the urinary collecting system.


Chromosomal studies in cryptorchidism.

Chromosomal studies in cryptorchidism. The chromosomes of undescended tests from 13 boys were analyzed. In 12 of these patients the peripheral blood chromosomes were examined and in 7 the chromosomes of normal skin cultures were examined. In addition to standard techniques the chromosomes also were analyzed by G and Q-banding methods. We were unable to detect any significant abnormalities of either chromosome number of structure in the undescended testes. Thus, our investigation failed to demonstrate a chromosomal lesions that may be responsible for maldescent of the testes. Our investigation did not reveal any changes in the chromosomes of these undescended testes that could be interpreted as indicating a potentially malignant state.


The staged orchiopexy: a critical review of the literature. During the last 20 years reports have appeared describing successful results with a 2-stage orchiopexy for patients noted to have a short testicular artery on first exploration. A critical review of these reports indicates that accurate documentation of the phenomenon of spontaneously increasing spermatic cord length following surgical manipulation does not exist. It is suggested that a scientifically accurate prospective study of the staged orchiopexy is needed.


Failure to prove arenavirus infection among the small mammals from an endemic area of Korean hemorrhagic nephrosonephritis. In the light of recent knowledge on a complex of diseases caused by a new group of viruses, arenaviruses, virological studies largely directed toward small field mammals were undertaken during 1973-1974 aiming at etiological clarification of Korean hemorrhagic nephrosonephritis (KHNN). Specimens were collected in an endemic area of KHNN located north to northeast of Seoul. Virus isolation tests with 299 urine specimens and 131 mite pools recovered from small mammals and 14 acute stage sera from typical cases yielded negative results. Complement-fixation (CF) tests failed to detect antibodies against the antigens of Congo, lymphocytic choriomeningitis (LCM), Tacaribe, and Pichinde viruses among 366 small mammal sera. In addition, CF tests of 59 of the above sera against Apoi and Lassa virus antigens were negative. The results do not support the likelihood of an arenavirus being transmitted among Korean small field mammals, the overwhelming majority of which were Apodemus agrarius. A hypothesis that KHNN is caused by a virus of small field mammal origin was not proved within the technical limit of relatively unsophisticated methods employed herein.


Effects of benzodiazepines and pentobarbital on the evoked potentials in the cat brain. The sites of action of benzodiazepines, diazepam and ID-540 [7-chloro-5-(o-fluorophenyl)-1-methyl-1,3-dihydro-2H-1,4-benzodiazepin-2-one) on the central nervous system were examined and compared with those of pentobarbital using evoked potentials recorded on the limbic system and hypothalamus in the cat brain. Benzodiazepines affected the various neuronal connections of the intra-limbic, limbic-hypothalamic and midbrain-limbic systems; especially the amygdala (AMYG)-, ventromedial hypothalamus (VMH)- and central gray matter (SGC)-hippocampal (HIPP) evoked potentials were attenuated, whereas the AMYG-VMH, VMH-AMYG and the septum (SP)-VMH evoked potentials were facilitated. Pentobarbital selectively attenuated the SGC-, VMH- and AMYG-HIPP evoked potentials, or facilitated the VMH-AMYG and the SP-HIPP evoked potentials. Both benzodiazepines and pentobarbital affected three afferent hippocampal neuronal connections, areas of the reticulo-hypothalamic systems regulating hippocampal activity, while only benzodiazepines affected the neuronal influence of the amygdaloid and septal areas on the hypothalamus.


Effects of L-glutamine of acetylsalicylic acid or taurocholic acid-induced gastric lesions and secretory changes in pylorus-ligated rats under normal or stress conditions. An oral dosing of either acetylsalicylic acid (ASA) or taurocholic acid (TCA) to pylorus-ligated rats subjected to water-immersion stress produced severe damage to the gastric musoca in contrast to the irritation observed in non-stressed ones. The irritative activity of ASA or TCA on gastric mucosa under stress was dose-dependent. Stress itself (23 degrees C, 7 hr) did not induce any appreciable changes in gastric mucosa under stress wasic mucosa of rats. L-glutamine, given together with SAS or TCA, significantly prevented the potentiated development of SAS- or TCA-induced gastric lesions in stressed rats. L-glutamine also prevented in varying degrees the reduction of acid and increment of Na+ ion in gastric juice accumulated in stressed rats in response to ASA or TCA.


Studies on monoamine oxidase. XVIII. Enzymic properties of placental monoamine oxidase. Enzymic properties of partially purified monoamine oxidase (MAO) from human placenta were studied with tyramine, serotonin and benzylamine as substrates. The highest activity was obtained with serotonin and almost no activity was observed with benzylamine. These results are similar to those obtained with rat placental MAO, but different from those with rabbit placental MAO. The Km values for serotonin and tyramine were found to be 0.21 mM and 0.23 mM, respectively and the pH optimum was 8.1 with either substrate. The thermal inactivation curves of this enzyme with the two substrates were identical. The pI curves for inhibition of MAO activity by harmine, pargyline and iproniazid were similar and almost the same pI 50 values for the respective inhibitors were obtained with the two substrates. MAO in human placenta differs from that in other organs, such as liver, brain and plasma from the standpoint of the substrate specificity and the inhibitor sensitivity. The possibility that human placenta contains a single form of MAO is discussed on the basis of the present results.


Antagonizing effects of beta-adrenergic blockers on lucus coeruleus-induced inhibition of trigeminal nucleus neurons. Effects of alpha- and beta-adrenergic blocking agents applied into the lateral ventricle were studied on the relay neuron in the rostral part of spinal trigeminal nucleus (STN) of cats. Conditioning stimulation of the locus coeruleus (LC) and sensory cortex (SC) inhibited the orthodromic spike generation in STN relay neuron without affecting the antidromic spike, as already reported, and re-confirmed herein. The LC-induced inhibition of orthodromic spike was significantly reduced by intraventricular administration of beta-blockers, MJ 1999 (5 mg) and propranolol (0.5 mg), while the SC-induced inhibition of orthodromic one was not modified by the beta-blockers. The antidromic spike in STN relay neuron per se remained unaffected by these treatments. Intraventricular administration of alpha-blockers such as phentolamine and phenoxybezamine produced no alterations of the LC- and SC-induced inhibition of orthodromic spike. As the beta-blockers produced a selective antagonism, noradrenaline originating in LC probably acts as an inhibitory transmitter on the STN relay neuron and is mediated by beta-receptor.


Reversible adrenergic alpha-receptor blocking action of 2,4'-dimethyl-3-piperidino-propiophenone (tolperisone). The vascular action of 2,4'-dimethyl-3-piperidino-propiophenone hydrochloride (tolperisone hydrochloride), a centrally acting muscle relaxant, was investigated in pentobarbital anesthetized dogs. Tolperisone given intravenously produced a transient hypotension, tachycardia, and hyperventilation. The drug increased the femoral arterial flow, and decreased the superior mesenteric arterial flow following an initial transient increase. When injected directly into femoral and mesenteric arteries, tolperisone caused a rapid increase in both arterial flow (vasodilatation). However, femoral vessels were about 90 times as sensitive as mesenteric vessels to tolperisone. These results indicate that tolperisone shifts the blood volume from mesenteric (visceral) vessels to femoral (skeltal) ones. The femoral vasodilatation produced by i.a. tolperisone was not depressed by the pretreatment with i.a. propranolol, atropine or chlorphenylamine. Tolperisone decreased the contractile force in an isolated and cross-circulated papillary muscle. Tolperisone produced adrenaline reversal and antagonized the pressor response to noradrenaline. Moreover, femoral vasoconstriction caused by i.a. adrenaline was converted to vasodilatation and that caused by i.a. noradrenaline was depressed during an i.a. infusion of tolperisone. These results indicate that tolperisone blocks adrenergic alpha-receptors. The blocking action was rapid in onset, short-lived, and in addition, competitive.


Pharmacological studies on triazine derivatives V Sedative and neuroleptic actions of 2-amino-4-[4-(2-hydroxyethyl)-piperazin-1-yl]-6-trifluoromethyl-s-triazine (TR-10). Pharmacological properties of 2 amino-4-[4-(2-hydroxyethyl)-piperazin-1-yl]-6-trifluoromethyl-s-triazine (TR-10) were investigated in mice and rats. Chlorpromazine served as a reference compound. Tr-10 expressed in general the pharmacological profiles as neuroleptic ascertained by anti-methamphetamine activity, supression of conditioned avoidance response, taming effects, decrease in exploratory behavior and cataleptogenic activity. Among these effects, anti-methamphetamine action was most potent. Different from chlorpromazine, TR-10 showed a similar pharmacological activity pattern in the intraperitoneal and oral routes of administration as depicted from ED50/LD50 values. Although the effects relevant to neuroleptics were less potent than chlorpromazine, such were seen with TR-10 at lower doses than those causing muscle relaxation. TR-10 significantly depressed the spontaneous motor activity but showed no anti-convulsant action in mice. Hypothermic action, potentiating effects of hypnotics and alpha-adrenergic blocking action, characteristic to chlorpromazine, were very weak for TR-10. TR-10 also showed low toxicity in mice (LD50 = 820 mg/kg p.o., 465 mg/kg i.p.) compared with that of chlorpromazine (LD50 = 370 mg/kg p.o., 228 mg/kg i.p.).


Effects of narcotic analgesics on serotonin metabolism in brain of rats and mice. The effects of narcotic analgesics on the brain 5-hydroxytryptamine (5-Ht) and 5-hydroxyindoleacetic acid (5-HIAA) levels of rats and mice were investigated in relation to our preceding data on the effect of humoral modulatorents. The results suggest that morphine accelerates the release of brain 5-HT both in rats and mice, and that neither methadone nor pethidine alters the brain 5-HT and 5-HIAA levels in rats. The morphine-induced increase in brain 5-HT turnover is likely to be involved in the morphine-induced decrease in locomotor activity and hypothermia in rats. The activity-decreasing effects of methadone or pethidine, on the other hand, are mediated by mechanisms different from those which mediate the effects of morphine. In contrast, an increase in brain 5-HT turnover in mice apparently does not play an important role on activity-increasing effects of morphine but rather participates in other pharmacological effects of morphine.


Selective alpha-adrenoceptor blocking actions of a new derivative of 2-halogenotheylamine: 6-(2-bromoethyl)-10,11-methylenedioxy-5,6,7,8-tetrahydrodibenz[c,e]azocine. A new compound, 6-(2-bromoethyl)-10,11-methylenedioxy-5,6,7,8-tetrahydrodibenz [c,e] azocine (DA-VIII-MBr) was found to have a more selective alpha-adrenergic blocking action than dibenamine or phenoxybenzamine. From dose-response curves for adrenaline and 5-hydroxytryptamine (5-HT) obtained in strips of rat aorta before and after incubation with each of the three blocking agents, the fractions of receptors remaining active for adrenaline and 5-HT, respectively, were estimated. After blockade with DA-VIII-MBr the receptors for adrenaline were blocked considerably, but those for 5-HT were little affected. Dibenamine blocked the receptors to adrenaline and 5-HT almost equally. The effective dose of phenoxybenzamine for adrenaline receptors was less than one hundredth that of dibenamine or DA-VIII-MBr, but specificity for these receptors was intermediate between those of dibenamine and DA-VIII-MBr. The structure of DA-VIII-MBr is an analog of apogalanthamine and its nitrogen atom bears the 2-halogenoethylamine group in part of an eight membered ring.


[Arteritis of the large vessels originating at the aorta (Takayasu arteritis) in childhood (author's transl)]. A 9-month-old female child with extensive inflammatory lesions in the large arterial trunks originating at the aorta (Takayasu arteritis) was reported. Severe cerebral, intestinal and coronary circulatory disturbances as well as insufficient blood supply in both arms, particularly the left arm, developed as a result of the stenoses and obliterations. Clinical course, postmortem findings and the results of EKG, EEG and carotid angiography were discussed. The onset in infancy and the very rapid progression of the disease were unusual.


[Cyclic AMP and plasma renin activity in renal vein blood after amitryptiline, theophylline, furosemide and beta adrenergic blocking substances (author's transl)]. The influence of amitryptiline, theophylline and furosemide on the concentration of cyclic-AMP and plasma renin activity (PRA) was investigated in renal vein plasma. Additionally, the stimulating effect of furosemide on the PRA after application of the beta-adrenergic receptor antagonists propranolol and practolol and the cyclic AMP concentration in the plasma were measured. All drugs were given intravenously. After amitryptiline cyclic-AMP concentration increased about 1.5-fold compared with the basal value,PRA was not altered. After theophylline cyclic-AMP concentration increased about 1.2-fold, PRA 2.0-fold compared with the basal value, PRA was not altered. After theophylline cyclic-AMP concentration increased about 1.2-fold, PRA 2.0-fold coa increased within 7 min and no further increase was observed till the 15th min. After practolol cyclic-AMP concentration and PRA decreased about 20% compared with the basal value within 10 min. The stimulating effect of subsequently applied furosemide on PRA was not altered, but the cyclic-AMP concentration was not changed in this time by furosemide. After propranolol cyclic-AMP concentration and PRA decreased about 20% compared with the basal value. The cyclic-AMP concentration was not influenced by the following furosemide application, in agreement with the findings after practolol, however, PRA could be stimulated only in 36% of these patients under beta-receptor blockade. Our results show that changes of the concentrations of cyclic-AMP and of PRA are independent of each other. An elevated intracellular cyclic-AMP level due to the inhibition of phosphodiesterase is as a single factor unable to stimulate renin release. Our results give no evidence of a direct involvement of the adenylcyclase-system in the mechanism of renin release. The effect of propranolol and practolol on the basal value of PRA and cyclic-AMP is equal. The different influence of 10 mg propranolol and 20 mg practolol on the stimulating effect of 40 mg furosemide on the PRA can be interpreted as a dosage problem.


[The effect of surgery, performed under ischemia, on the fine structure of the hand muscles]. The ultrastructural picture of the intact and injured hand muscles has been examined by the authors during and after operations in tourniquet ischaemia of 12 patients, in connection with tendon and nerve reconstruction. Muscle specimina were taken partly in the beginning of the ischaemization, and partly at the end ot it, every 15--20 minutes, and the muscle specimina have been examined by means of electron microscope. It has been found that in the first 2 hours of ischaemia neither the intact muscles, nor those previously injured suffer demonstrable damage.


[Simultaneous occurrence of Perthes' disease, spondylolysis-spondylolisthesis and other forms of aseptic necrosis]. Cleft formation on the lumbo-sacral spine has been observed by the authors in 15% of 32 patients with Perthes' disease--in whom follow-up examination has been performed without selection. In one case Perthes' disease was concomitant with spondylolisthesis causing no complaints. In 2 cases aspetic necrosis with other localization was found. It is presumed by the authors that between Perthes' disease and the arch rupture of the vertebrae causal relationship may exist--but also simultaneous occurrence with sterile bone necrosis with other localization is to be taken into consideration.


[Axillary block anesthesia in hand surgery]. In spite of the use of narcosis on a large scale local anaesthesia is necessary in the surgery on the hand. After the discussion of the possibilities and types of local anaesthesia the axillary blocking of the plexus brachialis is dealt with in detail, and on the basis of the experience gained with 150 cases the advantages of this method are evaluated.


Thermodynamic parameters for the intramolecular disordered-to-beta transition of poly(L-tyrosine) in aqueous solution. We have examined the disordered-to-beat transition of two samples of high molecular wieght poly(L-tyrosine), a polypeptide with an ionizing side chain, by potentiometric titration of solutions of this polymer in aqueous buffer. The degree of ionization was determined spectrophotometrically from the absorption of the tyrosinate anion at 293.5 nm. By using polymer solutions of high dilution the region of the conformational transition was separated from that of aggregation. As a consequence it was possible to obtain titration data for the intramolecular transition which we believe are free of interference from aggregation. For this transition we found that deltaHO=-1330 cal/mol of residue. The two samples gave values of -3.42+/-0.14 and -3.85+/-0.10 u for the change in entropy. Possible artifactual origins for this distinction have been examined and may be dicounted. Rather, it probably reflects a difference in the beta structures formed which depends on molecular weight.


[Substitution of skull defects with methyl acrylate]. On the basis of the data of the literature and their own experiences the theoretical and pratical problems of cranioplasty are dealt with by the authors. The indications of the cranioplasty and the various materials employed are discussed. In the author's opinion, the metacrylate polymerizating to heat has been found the most practical. The use of the metacrylate implant, the modelling and the preparation of the implant, as well as a new method worked out by the authors are discussed. The operative technique, the results and the possible complications are dealt with. Also the authors' new method in order to prevent the infection from the nasal cavity is described.


[Collagen synthesis, studied by sub microscopic methods in beta-propiolactone and in tendon homografts, preserved by gamma irradiation]. In dog experiments homologous tendos conserved in beta-propiolacton and by means of gamma-radiation have been transplanted by the authors. After the 1st, 3rd and 8th post-operative week various regions of the graft and the recipient tendon stump have been examined by means of light--, polarisation microscopic and electron microscopic method. The examinations have unanimously shown that the tendon grafts conserved in beta-propiolacton are reorganized quickly, within 8 weeks nearly completely besides hardly visible inflammatory reaction. In the meantime, the originally collagen fibrous framework is decomposed and on it a new collagen fibrous structure develops, and thus a new tendon--capable to function--is formed. On the other hand, the reorganization of the grafts conserved with gamma-radiation is protracted or it fails to occur, and cicatrization is observed. Some problems of the modern tissue preservation, the collagen synthesis occurring in the active fibroblasts, the secretion of the collagen precursory substance into the extracellulary space, as well as the formation, the structure and the constitution of the newly synthesized collagen fibres--forming the transplant--are discussed by the authors. The obtained results are demonstrated in light-, polarization microscopic and electron microscopic pictures.


[Surgical treatment of chemical injuries]. Among nearly 2000 patients of a traumatological ward 82 patients were treated with chemical injury during 6 years. On the basis of their experiences the tissue damaging effect of the chemical substances as well as the methods of treatment are discussed by the authors. Dermoplastic operations have been performed in 28 injured patients. Two patients died--the immediate cause of death was in both patients pneumonia developed as complication of important skin defect. As first-aid abundant sponge-bath and flushing are recommended by the authors--the neutralizing drugs play a secondary role. In the case of deep injuries primary necrectomy is recommended, firstly in colliquation necrosis, as well as in the cases, in which the absorption of the poisonous substance is to be feared of.


[Intramural duodenal hematoma of traumatic origin]. A rare case of the blunt abdominal injuries--forming a clinical entity--intramural duodenal haematoma is reported. The symptoms of IMDH may appear--non specifically--after a longer interval. The diagnosis is assured in the first place by careful anamnesis and on the basis of the characteristic radiological signs. The treatment may be--depending on the extent of the obstruction--conservative or surgical--in the latter case mostly the evacuation of the haematoma is performed.


[Acute traumatic infratentorial subdural hematoma]. The case of an injured man, aged 54, is reported. On the basis of the neurological symptoms, without neuro-radiological contrast examinations, in presence of negative skull osteogram, on the day of the injury suboccipital craniectomy and removal of the subdural haematoma have been performed. The diagnostical difficulties of the occipital injury are pointed out, since both the occipital and infratentorial regions are "silent" regions for the carotis-angiography. In the case of subdural haematoma the suboccipital decompression performed in due time may be exactly as much successful as the supratentorial decompression.


[Mistakes, errors and their prevention in the medical expertise about accidental injuries]. It may be stated that in the examination of the individuals who had suffered an accident, as well as in the assessment of the subsequent consequences--for whatever authority (social assurance, court of justice, State Insurance Company) the medical opinion is given--various errors may be committed, which may direct the claim for indemnity in a faulty way and may endanger also the medical competence of the expert opinion. The medical, social, legal and economical relationships of these errors have repeatedly been pointed out by the authors. Neither the patient nor the community may suffer a loss or injury merely by the reason that the medical expert's skill was defective regarding the medical expert's work. The aim of the authors' paper is to through a flash of light on these errors and mistakes and possibly to improve the work of the medical experts in these special fields.


Immunohistochemical studies on the localization and distribution of monoamine neuron systems in the rat brain. I. Tyrosine hydroxylase in the mes- and diencephalon. The localization and distribution of tyrosine hydroxylase (TH), the first enzyme in the catecholamine synthesis, in the mes- and diencephalon has been studied with the indirect immunofluorescence technique of Coons and collaborators. Principally, TH was present in neuron systems with a distribution similar to known dopamine, noradrenaline and adrenaline systems. The present data, taken together with published and some unpublished results, indicate that all parts of most central dopamine neurons, i.e. cell body, dendrites, axon and nerve terminals, appear strongly fluorescent. The adrenaline neurons also appeared strongly fluorescent, except for their axons, which only exhibited a weak fluorescence. Only cell bodies of noradrenaline neurons were strongly fluorescent, whereas the nerve terminals and axons showed a weak or moderate fluorescence intensity. The fine noradrenaline nerve terminals in some areas, such as the thalamus, were invisible or, under favourable conditions, weakly fluorescent. Therefore, in the present study we are mainly dealing with the dopamine neurons of the upper brain stem. Our results demonstrate a widespread occurrence of TH-positive neuron systems in the mes- and diencephalon. The different mesencephalic dopamine systems and their ascending projections were visualized. Numerous TH-positive cell bodies were present along the ventricle system extending from the aqueductus cerebri to the most cranial periventricular parts of the third ventricle. The caudal part of these neurons, consisting of very small cell bodies, belong to the dorsal periventricular system described by Lindvall and Björklund. Several TH-positive cell bodies were also observed in the inferior collicle of young animals. In the superficial layers of the inferior collicles TH positive nerve terminals were seen. At the hypothalamic level the A11 to A14 cell groups as well as some additonal cell bodies and extensive nerve terminal plexuses appeared strongly fluorescent. The differences in the intensity of the TH-related immunofluorescence between various brain regions and between various neuron systems may well reflect differences in enzyme levels between the various catacholamine systems rather than be due to the existence of different types of TH.


A study of physicians' assistants in a rural setting. In an effort to improve the health care in a rural county in Southern Appalachia, physicians' assistants (MEDEX) have been employed in the offices of three general practitioners over a three-year period. This program was evaluated using a before-after and experimental-control (county) design, utilizing data both from physician office contracts and a continuing survey of the populations of the experimental and control counties. Results show the following: 1) utilization (average office visits per week) increased; 2) types of care (preventive versus curative) remained stable; 3) the hospitalization rate increased continuously during the three years for those physicians using physicians' assistants; and 4) the physicians' assistant functioned more as a physician substitute than as an assistant. It is concluded that although use of physicians' assistants may increase utilization rates, they may not reduce the long-range cost of medical care through providing more preventive or ambulatory (as opposed to hospital) care.


Adding a Medex to the medical mix: an evaluation. Three classes of Medex were followed during their preceptorship training and subsequent employment to assess the impact of adding such physician assistants on 13 private practices. The data suggest that the Medex are readily incorporated into the practice, seeing a similar spectrum of patients and generally functioning as a semicolleague. Although the Medex was almost universally a contributing addition to the practice, increasing patient volume, the specific effects varied with the setting.


[Alternativity of methane assimilation pathways in obligate methylotrophs]. The activity of key enzymes involved in the primary pathways of methane assimilation and enzymes of the citrate cycle was determined in various obligate methylotrophs: mesophilic, thermotolerant, and thermophilic. The bacteria are characterized by the membrane ultrastructure of the I type, high activity of hexosephosphate synthase, NAD- and NADP-specific isocitrate dehydrogenase, and the absence of alpha-ketoglutarate dehydrogenase. The bacteria also displayed the activity of key enzymes of the serine cycle, hydroxypyruvate reductase, and serineglyoxylate aminotransferase. Therefore, both the ribulose monophosphate and serine pathways are involved in methane assimilation, and the division of methanotrophs into two groups, according to their metabolism, is tentative.


[Effect of H+, OH-, Cu2+ and Ag+ on amino acid composition of Candida utilis cells in chemostat culture]. Amino acid composition was determined in the cells of Candida utilis VKM Y-1668 growing in the regime of chemostat. The growth was inhibited by pH of the medium, copper and silver ions, and glycerol deficiency as a control. Some specific differences have been found in the amino acid composition of the yeast depending on the nature of the inhibitor.


[Biosynthesis of chitinase by Achromobacter liquefaciens]. Bacterial cultures under study synthesize exocellular chitinase on a medium containing chitin or demineralized crab shells as a source of carbon and nitrogen. Conditions for biosynthesis of chitinase by the cells of Achromobacter liquefaciens 301a were investigated under periodic and continuous conditions of cultivation. The preparation of chitinase isolated from the cultural broth of A. liquefaciens 301a hydrolysed colloid and native chitin at the optimum pH 6.5 and temperature 40degreesC. The terminal products of the reaction are chitobiose and N-acetylglucosamine.


The physician's assistant. A controversial topic at home and abroad. Non-physician primary care providers (physicians' assistants) have existed in under-developed or emerging nations, in various forms, for many years. It is only recently, however, that this concept has been introduced into countries with more advanced economic and medical care systems. An active and rapidly developing physician's assistant movement has been organized in the United States since the early 1960s; physician's assistants and nurse practitioners may now be found in the almost every one of the 50 United States. Recently, this kind of concept has been suggested as a possibility for improving the Australian medical care system. This article explores the history and present status of the physician's assistant/nurse practitioner movement in the American medical care scene.


[Cultivation conditions of the Aspergillus terricola mutant producing proteolytic enzyme]. The effect of temperature, aeration and various foam suppressors on the growth of a mutant of Aspergillus terricola H-20 and on its production of proteolytic enzymes was studied. A high level of the proteolytic activity (12-15 PU/ml) was found at 28-30 degree C in conditions of aeration with the rate of oxygen dissolution being 0.38-0.86 g of oxygen per litre per hour. Synthesis of the enzyme was inhibited by a decrease in temperature to 22-24 degree C, a surplass (1.07 g O2 per litre per hour) or insufficient (0.18 g O2 per litre per hour) content of oxygen in the medium, and by addition of animal (sperm-whale) fat as a froth breaker to the medium. The best rate of protease biosynthesis was obtained upon stepwise addition of sunflower oil to the cultural broth. The best synthetic froth breakers for biosynthesis of the proteolytic enzyme are adecanol at a concentration of 0.1-1% and polymethylsiloxane PMS-A 154 as a water emulsion at a concentration of 0.5% of the medium volume.


[Effect of growth medium composition of glucoamylase and glycosyltransferase activity of Endomyces fibuliger]. Production of glucoamylase and glycosyltransferase by Endomyces fibuliger was found to depend on sources of carbon and nitrogen nutrition. Starch at a concentration above 0.5% in the medium stimulated biosynthesis of glycosyltransferase but inhibited production of glucoamylase by End. fibuliger 20-9. The rate of growth of the micro-organism increased by a factor of 3.3 with an increase of starch concentration from 0.5 to 6%. Synthesis of glycosyltransferase was repressed by glucose, lactose, sucrose and maltose. Synthesis of glucoamylase was repressed by lactose, sorbose and galactose. Synthesis of glycosyltransferase was stimulated by xylose, sorbose and galactose. Production of glucoamylase was stimulated by xylose and arabinose. Growth of the culture and synthesis of glucoamylase and maltase in the cultural broth were stimulated by an increase in the concentration of maize extract. Biosynthesis of glucoamylase and glycosyltransferase was stimulated by NH4H2PO4.


[Isolation and properties of the fibrinolytic enzyme from the Actinomyces thermovulgaris cultural broth]. A fibrinolytic enzyme was isolated from the cultural broth of Actinomyces thermovulgaris T-54 by precipitation with ammonium sulphate to 0.8 saturation and chromatography on GE-cellulose and CM-cellulose. The enzyme is homogeneous as was confirmed by disk electrophoresis in polyacrylamide gel. Some properties of the enzyme were studied. A modification of the method is suggested to assay the fibrinolytic activity.


New approaches to the causation and prevention of cancers of epithelial surfaces. Environmental rather than genetic factors appear to be responsible for the causation of cancers involving epithelial surfaces, especially oropharyngeal cancers, cancers of the gastro-intestinal tract, cancers of the liver and gall-bladder, cancers of the cervix uteri and lung cancers. Epithelial surfaces which have mucus-bearing cells are particularly prone, because if these mucus cells are surrounded by an alkaline milieu over a prolonged period, the mucous of the epithelial cells is rendered fluid and is removed from the cells, of which it forms an integral part. This leads to proliferative changes with metaplasia, cell-atypia and a very significant increase in mitotic activity. Such changes are a prelude to carcinogenesis. In the case of many oropharyngeal cancers in Asia and Africa it is the alkaline slaked lime in the betel quid which is responsible; in the case of gastric cancers it is the reflux of the alkaline duodenal contents into the stomach; in the case of colon it is the absence of roughage, cellulose and vegetable fibres from the diet; in the case of the cervix uteri it is the frequency of coitus which gives rise to a highly alkaline seminal fluid; and in the case of cancer lung it is the alkaline cigarette smoke.


Application of halide ion nuclear magnetic resonance to bioinorganic problems. The basis for the application of halide ion nuclear magnetic resonance to the investigation of biochemical problems is reviewed and a summary of applications is presented. Anion binding to macromolecules, low molecular weight or model compounds, metalloenzymes, and extrinsic metal interactions with macromolecules are discussed.


Determination of intermediary metabolites in yeast. Critical examination of the effect of sampling conditions and recommendations for obtaining true levels. The effect of sampling conditions on the levels of adenine nucleotides, pyridine nucleotides, glycolytic intermediates and related metabolites in yeast has been studied. A systematic examination of the conditions for harvesting has shown that it can be best accomplished by rapid filtration. Delays in the handling for removal of the medium, as is usual in the process of obtaining a number of data reported in the literature, lead to important changes in some of the metabolites examined. It is also shown that when a washing is imperative it can be carried out with a methanol-water mixture (50/50, v/v) cooled at -40 degrees without loss of intracellular concentrations of non-readily diffusible metabolites. On the basis of this experience the outline of a generally applicable procedure is presented.


The extracellular metalloprotease of Serratia marcescens: I. Purification and characterization. An extracellular protease of Serratia marcescens produced during growth on skim milk medium was isolated by ethanol precipitation. The protease was purified by salt fractionation, DEAE-cellulose ion exchange chromatography and gel filtration chromatography on Agarose P-100. It has a broad optimum from pH 6.0 to 9.0 and a temperature optimum of 45 degrees C for proteolytic activity on casein. It was classified as a metallo-protease by virtue of its inactivation by metal-ion chelators and reactivation by ferrous ions. Proteolytic activity was not affected by diiso-propylfluorophosphate, p-chloromercuribenzoate and dithiothreitol.


[Juvenile rheumatoid arthritis and related collagen diseases. Clinical aspects (author's transl)]. Juvenile rheumatoid arthritis or, more correctly, juvenile chronic polyarthritis with its many clinical manifestations can be separated into the Still-syndrome with acute beginning, high fever and a high percentage of extra-articulalar, i.e. visceral symptoms, and the chronic polyarthritis in the more strict sense with non-visceral symptoms. The subsepsis allergica should be regarded as a subseptic first stage of the Still syndrome. The Still-syndrome implies a systemic disease mainly of the reticulo-endothelial system, with carditis, nephropathy, recurrent erythemas, and a progressing polyarthritis. Later symptoms are amyloidosis, chronic nephritis, myo- and pericarditis, and artheriitis necroticans. Predominanly the involvement of the kidneys is the reasons for the high mortality rate of 13%. Chronic polyarthritis in the strict sense is similar in children and adults, though in children rheumatic factors are rarely detected. The exsudative form of arthritis tends to cause early deterioration. Joint symptoms are distributed asymmetrically and show locally inflammed growth otherwise less common in Still-syndrome. Spondylitis cervicalis rapidly causes ankylosis. Atlanto-axial-arthritis with consequent atlanto-axial dislocation can be the reason for neurological disturbances. Juvenile mono- or oligo-arthritis often turns into polyarthritis; but for joints the prognosis is more favourable. In contrast, rheumatoid iridocyclitis as found in 22% of the cases causes unfavourable complications because symptoms are not noticed in time so that treatment is often too late. Juvenile spondylitis ankylosans begins with a peripheral arthritic stage which is not easily distinguished from chronic polyarthritis. The male sex, mono- or oligoarthritis of the outer extremities, pain in the heel, atlanto-axial-arthritis, iridocyclitis, and a positive HLA of 27 give a diagnostic clue. -- Characteristics of the therapy will be discussed.


[The ecology and medical importance of the Tahyna Virus (author's transl)]. In 1958 a virus transmitted by a mosquito was isolated in the Slovakian village of Tahyna. Since this virus was unknown in Europe, a detailed study was begun. It was shown that this virus belonged to the California group of arboviruses and occurs in practically all European countries. It could very well be that young hares are the hosts for this virus, and that it multiplies in them. Foals and suckling pigs are also hosts and possibly also increase the virus. In patients observed so far, infection with the Tahyna virus appears with influenza-like symptoms. In some cases, meningoencephalitis and atypical pneumonia were observed.


Formation of mutagenic N-nitroso compounds from the pesticides prometryne, dodine and carbaryl in the presence of nitrite at pH 1. Environmental chemicals including pesticides carrying secondary and tertiary amino groups are suggested to be a health hazard to man since potentially carcinogenic nitroso compounds may be formed in the presence of nitrite at low pH values resembling conditions in the human stomach. Nitrosation of the isopropylamino-triazine Prometryne, the n-dodecyl guanidine Dodine and the N-methylcarbamate carbaryl was investigated in the presence of HCl and acetic acid at pH 1 and excess sodium nitrite for 4 h at 37 degrees C. The reaction products were extracted with CCl4 and were analyzed qualitatively and quantitatively by infrared spectroscopy, nuclear-resonance spectrometry, GC/mass spectrometry and by spectrophotometry. All compounds investigated formed N-nitroso derivatives in the following yields: carbaryl 67%, Dodine 12% and Prometryne 14%. The N-nitroso derivatives per se were not or only slightly mutagenic to Escherichia coli K12 or Salmonella typhimurium TA 1538. However, significantly increased mutation frequencies were seen after metabolic activation by mouse-liver microsomes. These results add to the observations that among environmental chemicals not only those containing methyl- or ethyl-substituted amino groups form potentially carcinogenic nitroso derivatives but also those with iso-propylamino groups as well as alkyl-substituted guanidine derivatives.


Thyrotropin releasing hormone (TRH): restoration of oxotremorine tremor in mice. Comparison with quipazine, a serotoninergic and dopaminergic stimulant. Both TRH and quipazine (2.5-25 mg/kg) were found to restore and to intensify the oxotremorine-induced tremor in mice when injected i.p. 60 min after oxotremorine 0.5 mg/kg s.c. This phenomenon does not seem to be due to an increase in body temperature or muscle tone. Also other dopaminergic drugs, e.g. amphetamine, methylphenidate, nomifensine and apomorphine had a significant but lesser effect than TRH or quipazine. Haloperidol and methysergide both antagonized the effect of quipazine but not that of TRH. Neither propranolol, phenoxybenzamine, alpha-methyl-p-tyrosine, nor p-chlorophenylalanine inhibited the activity of TRH or quipazine. The restoration of oxotremorine-induced tremor could be prevented by atropine but not by methylatropine. It is concluded that quipazine exerts its effect by direct stimulation of serotoninergic and dopaminergic receptors, whereas TRH receptors may represent separate entities and TRH may function as a neurotransmitter or neuromodulator.


Interaction of benzodiazepines with neuroleptics at central dopamine neurons. Several benzodiazepines (chlordiazepoxide, clonazepam, diazepam and flunitrazepam) markedly counteracted the elevation of the homovanillic acid (HVA) content of the rat brain induced by neuroleptics (haloperidol, pimozide, chlorpromazine, and clozapine). A similar effect was obtained with the inhibitor of GABA transaminase, aminooxyacetic acid (AOAA). The interaction of benzodiazepines with the neuroleptic-induced HVA increase was similar in the striatum and in the limbic forebrain and was antagonized by the GABA receptor-blocking agent, picrotoxin. Both the benzodiazepines used and AOAA potentiated the cataleptic effect of the four neuroleptics. It is concluded that benzodiazepines, by intensifying GABA-ergic transmission, enhance the ongoing inhibition of mesencephalic dopamine neurons exerted by the striatonigral GABA system. As a consequence, the feedback activation of dopamine neurons induced by the neuroleptic blockade of dopamine receptors in the striatum and the limbic system is attenuated. This results in a reduction of the neuroleptic-induced increase of HVA and in the potentiation of the cataleptic effect of neuroleptics.


Effects of enzyme induction and inhibition on microsomal oxidation of 1,4-benzodiazepines. Metabolic oxidative profiles of diazepam (I) were obtained by aromatic C-4'-hydroxylation, N-1-demethylation, and 3-hydroxylation using a supernatant of rat liver. Incubation of 3-methyldiazepam (VI), which suppressed 3-hydroxylation, and N-1-nor-3-methyldiazepam (VII), were used to separately investigate these three oxidative pathways. Treatment of animals with phenobarbital enhanced N-1-demethylation and 3-hydroxylation, and to a variable extent C-4'-hydroxylation. Application of metyrapone reduced metabolite formation by 3-hydroxylation and N-1-demethylation, but had no effect on C-4'-hydroxylation. Metyrapone inhibition was more pronounced following than prior to phenobarbital treatment. C-2-hydroxylation was studied using medazepam (XX) incubations. This pathway was increased by phenobarbital pretreatment and reduced by metyrapone inhibition which was again more pronounced following than prior to phenobarbital pretreatment. These results support earlier conclusions on the heterogeneity of liver microsomes and suggests the presence of different species of hepatic microsomal terminal oxidases. Phenobarbital treatment and metyrapone change the metabolic profile via induction and inhibition, respectively, and, thus, in the case of 1,4-benzodiazepines, the formation of metabolites with varying pharmacological activity. This could become important in clinical situations as a diagnostic mean to determine induction under various treatment or, possibly, during cumulation of metabolites with a long half-life.


Restoration by histamine of the calcium-dependent electrical and mechanical response in the guinea-pig papillary muscle partially depolarized by potassium. Electrophysiological and mechanical effects of histamine were observed in guinea-pig papillary muscle which had been depolarized and rendered inexcitable by elevation of potassium concentration in Tyrode solution to 27 mM. 1. Histamine (3 X 10(-7) to 3 X 10(-5) M) restored the action potential and tension development. The amplitude of the action potential was increased by 31.6 mV/10-fold increase in extracellular Ca2+ concentration. Nifedipine (10(-6) M) abolished the electrical and mechanical responses which had been restored by histamine (10(-5) M) but TTX (10(-5) M) did not affect them. Reduction of the extracellular Na+ concentration to one half decreased the amplitude and the maximum rate of rise of the action potential restored by histamine (10(-5) M) while the peak tension was increased and an after-contraction occurred. 2. The maximum rate of rise and the amplitude of the action potential restored by histamine (10(-5) M) decreased with increase in stimulus frequency from 0.1-1.6 Hz. The peak tension decreased and then increased. The shape of the developed tension was also changed. In the presence of caffeine (1 mM), the only effect of an increase in stimulus frequency was a decrease in peak tension but the change in the shape of developed tension did not occur. 3. The electrical and mechanical responses restored by histamine (3 X 10(-6) or 10(-5) M) were depressed by metiamide (3 X 10(-6) M) but not by diphenhydramine (10(-5) M) or bufetolol (10(-6) M). 4. The electrical response restored by histamine (10(-6) or 10(-5) M) was enhanced by papaverine (10(-5) M) and depressed by N-methylimidazole (10 mM). It is concluded that histamine may enhance the slow inward Ca2+ current mediated by histamine H2-receptors and the adenylate cyclase system in ventricular muscle and that the positive inotropic action of histamine may be attributed to these mechanisms.


Inhibition by locally applied alpha-adrenoreceptor blocking drugs of the depressor response to stimulation of the anterior hypothalamus. Cats were anaesthetized with pentobarbital sodium and the anterior hypothalamus was superfused with artificial cerebrospinal fluid through a push-pull cannula. Electrical stimulation of the superfused area with the tip of the cannula elicited a fall of the arterial blood pressure which was dependent on frequency and voltage. Maximal depressor response was obtained at 60 Hz with 2-4V; further increase of the voltage often led to a rise of the arterial blood pressure. Superfusion of the anterior hypothalamus with the alpha-adrenoreceptor blocking drugs tolazoline, piperoxan, yohimbine or phentolamine caused a dose-dependent inhibition of the depressor response to hypothalamic stimulation. Tolazoline was less effective than the other drugs. Superfusion of the anterior hypothalamus with the alpha-sympathomimetic drug phenylephrine prior to and during superfusion with phentolamine abolished the inhibitory action of the latter drug. It is concluded that alphaadrenoreceptors are present in the anterior hypothalamus and involved in the depressor response to electrical stimulation of this hypothalamic area.


Dopaminergic neurons: reversal of effects elicited by gamma-butyrolactone by stimulation of the nigro-neostriatal pathway. In vivo studies demonstrate that administration of gamma-butyrolactone, a precursor of gamma-hydroxybutyric acid causes a rapid increase in endogenous levels of striatal dopamine and an increase in tyrosine hydroxylase activity measured by following the short term accumulation of dihydroxyphenylalanine. The increase in dopamine produced by GBL is blocked by stimulation of the nigro-neostriatal pathway. If dopamine is allowed to accumulate for 30 min following administration of GBL this increased dopamine can be released by stimulation of the nigro-neostriatal pathway. Maintenance of neuronal activity in the nigro-neostriatal pathway by continuous stimulation at a physiological frequency of 3/s effectively blocks the ability of GBL to cause an increase in tyrosine hydroxylase activity in the striatum on the stimulated side. Tyrosine hydroxylase activity in the non-stimulated contralateral striatum is increased over 100% by administration of GBL. Stimulation of the nigro-neostriatal pathway 30 min after GBL administration causes about a 500% increase in the accumulation of dihydroxyphenylacetic acid in the striatum on the stimulated side. These results suggest that the increased dopamine is present in a pool which is releasable by neuronal stimulation and is subsequently exposed to MAO. These results are also consistent with the hypothesis that GBL activates tyrosine hydroxylase and increases endogenous dopamine levels primarily by blocking impulse flow in central dopaminergic neurons.


The action of a toxin from the sea anemone Anemonia sulcata upon Mammalian heart muscles. The cardiac activity of toxin II, a basic polypeptide (m.w.: 4770) from the sea anemone Anemonia sulcata, was investigated in isolated electrically driven guinea-pig and rat auricles, Langendorff heart preparations of guinea-pigs and cat heart-lung preparations. Low concentrations of toxin II (2-100 nM) evoked a dose-dependent positive inotropic effect in the three different heart muscle preparations investigated. Higher concentrations of toxin II produced toxic symptoms like contracture and arrhythmia in auricles and atria (about 25 nM). In isolated cat hearts high toxin II concentrations (about 160 nM) caused unusual toxic symptoms such as long periods of ventricular fibrillation alternating with periods of normal cardiac activity. In rat and guinea-pig auricles as well as in Langendorff hearts of guinea-pigs the extent and rate of the positive inotropic effect induced by toxin II depended on the extracellular calcium concentration (0.45 to 2.7 mM). Toxin II did not alter the heart rate in spontaneously beating isolated cat hearts. In electrically driven guinea-pig auricles, the rate of the inotropic effect induced by toxin II was accelerated by increasing stimulation frequencies. Toxin II did not change the coronary flow in Langendorff heart preparations of guinea-pigs.