Properties and concentration of somatomedin A in various rat tissues. The concentration of somatomedin A (SMA) in various rat tissues was determined directly by the radioreceptor assay in supernates of tissue homogenates at 100,000 x g for 60 min. The SMA concentrations in pancreas, kidney, lung, liver, spleen, testis, brain and muscle were 2.56, 2.40, 2.28, 2.16, 1.56, 1.44, 0.60 and 0.30 U/g of wet tissue, respectively. The SMA content in rat serum was 6.66 U/ml. The SMA concentration in tissues never exceeded that in serum. When the 100,000 x g pellet of liver was treated with a hypotonic solution, the SMA content in the hypotonic solution was about 35% of that in the initial supernate. This result indicates that about 30% of the SMA in liver was stored in organellae. SMA in tissues decreased with incubation at 37 degrees C, though SMA in serum did not. This decrease could be precluded by adding 0.75 mM HgCl2 to the liver homogenate. The SMA content of the liver homogenate was dependent on growth hormone in experiments both in vivo and in vitro.


Degradation of somatomedin A by various organ homogenates from rats. Degradative activities of somatomedin A (SMA) have been examined in various tissue homogenates of rat using trichloracetic acid precipitable radioactivity of 125I-SMA. Kidney and testis showed higher specific activities and liver and brain lower activities. They were dependent on SH reagents; 0.5 mM HgCl2 inhibited the degradative activity of liver completely and 1 mM dithiothreitol (DTT) augmented the activity slightly. The activities in liver were separated by differential centrifugation; about 90 per cent of the total activity in the whole homogenate was recovered in the supernatant fraction at 100,00 x g for 60 min, and 10 per cent in the precipitate. The pH profile of each fraction was different; that of the supernatant showed a single peak at pH 7.4 and that of the pellet revealed two peaks at pH 5.9 and 7.4. However, both fractions showed similar SH-dependency.


The clinical evaluation of the simultaneous measurements of human chorionic gonadotropin (hCG) and its alpha-subunit in sera of patients with trophoblastic diseases. The concentrations of human chorionic gonadotropin (hCG) and its free immunoreactive alpha-subunit (hCG-alpha) in the sera of patients with trophoblastic diseases were measured by hCG and hCG-alpha radioimmunoassay (RIA), respectively. In the sera of 12 women with hydatidiform mole large amounts of hCG and considerably high level of hCG-alpha were detected in all cases. After the evacuation of mole the serum level of these glycoproteins decreased, the leve of hCG-alpha declined more rapidly than hcg. in the sera of patients with destructive mole the concentration of hCG-alpha was usually lower than that of hCG. After hysterectomy and chemotherapy the levels of hCG-alpha declined practically paralleling that of hCG. However, when hCG had decreased to undetectable level, hCG-alpha could no longer be detected in all cases. Although in the serum of patient with choriocarcinoma involving the uterus and lungs the concentration of hCG-alpha was almost as high as that of hCG, the secretory pattern of hCG and hCG-alpha might not be closely related. The changes in the serum level of free hCG-alpha as well as that of hCG parelled the clinical course of the patients examined in this study. The present results suggest that measurements of the serum free hCG-alpha may be a useful parameter to follow the clinical course and to evaluate the efficacy of treatments of trophoblastic diseases.


Post-partum transient thyrotoxicosis: report of two cases. Two patients with post-partum transient thyrotoxicosis associated with painless thyroiditis and low radioactive iodine uptake were described. The surreptitious use of thyroid hormones or iodine was excluded. Although the clinical course was compatible with that of subacute thyroiditis, passing through the hyperthyroid, euthyroid, hypothyroid and recovery phase, the patients showed a sustained elevation of serum anti-thyroglobulin antibody titer during the entire phase of the disease. Moreover, the histological findings obtained by the thyroid biopsy performed in a case were characteristic of chronic lymphocytic thyroiditis. Furthermore, it was of interest that the disease recurred following every delivery in 2 cases, suggesting the possible role of immunological changes induced by pregnancy and delivery in the etiology of this disease.


Serum estradiol and radial mineral content in postmenopausal females. The serum estradiol concentration and bone mineral content of the right radius were determined in 34 postmenopausal females. Regression analysis showed a significant positive correlation between the serum estradiol concentration and bone mineral content of the right radius (r=0.477, p less than 0.01). These results support the view that the decreased level of serum estrogens is one of the major factors involved in the loss of bone mass with age.


Measurement of forearm blood flow by venous occlusion plethysmography: influence of hand blood flow during sustained and intermittent isometric exercise. The requirement for using an arterial occlusion cuff at the wrist when measuring forearm blood flows by plethysmography was tested on a total of 8 subjects at rest and during and after sustained and intermittent isometric exercise. The contribution of the venous effluent from the hand to the forearm flow during exercise was challenged by immersing the arm in water at 20, 34, and 40 degrees C. Occlusion of the circulation to the hand reduced the blood flow through the resting forearm at all water temperatures. There was an inverse relationship between the temperature of the water and the proportion in the reduction of forearm blood flow upon inflation of the wrist-cuff, ranging from 45 to 19% at 20 degrees to 40 degrees C, respectively. However, during sustained isometric exercise at 10% of the subjects maximum voluntary contraction (MVC) there was no reduction in the measured forearm flow when an arterial occlusion cuff was inflated aroung the wrist. Similarly, there was no alteration in the blood flow measured 2 s after each of a series of intermittent isometric contractions exerted at 20% or 60% MVC for 2 s whether or not circulation to the hand was occluded nor of the post-exercise hyperemia following 1 min of sustained contraction at 40% MVC. These results indicate that a wrist-cuff is not required for accurate measurement of forearm blood flows during or after isometric exercise.


Electromechanical delay in human skeletal muscle under concentric and eccentric contractions. In contraction of skeletal muscle a delay exists between the onset of electrical activity and measurable tension. This delay in electromechanical coupling has been stated to be between 30 and 100 ms. Thus, in rapid movements it may be possible for electromyographic (EMG) activity to have terminated before force can be detected. This study was designed to determine the dependence of the EMG-tension delay upon selected initial conditions at the time of muscle activation. The right forearms of 14 subjects were passively oscillated by a motor-driven dynamometer through flexion-extension cycles of 135 deg at an angular velocity of approximately equal to 0.5 rad/s. Upon presentation of a visual stimulus the subjects maximally contracted the relaxed elbow flexors during flexion, extension, and under isometric conditions. The muscle length at the time of the stimulus was the same in all three conditions. An on-line computer monitoring surface EMG (Biceps and Brachioradialis) and force calculated the electromechanical delay. The mean value for the delay under eccentric condition, 49.5 ms, was significantly different (p less than 0.05) from the delays during isometric (53.9 ms) and concentric activity (55.5 ms). It is suggested that the time required to stretch the series elastic component (SEC) represents the major portion of the measured delay and that during eccentric muscle activity the SEC is in a more favorable condition for rapid force development.


Muscle strength and body composition as determinants of blood pressure in young men. The correlations of blood pressure to various indices of muscularity and fatness were studied in 183 young healthy men (mean age 19.7, SD 2.1 years). Systolic pressure showed significant positive correlations with body fat percentage, isometric strength of trunk extensors, body mass index, lean body mass, strength of leg extensors, heart rate, and the sum of four skinfolds. Diastolic pressure had significant positive correlations with body mass index, lean body mass, body fat percentage, sum of skinfolds, strength of leg extensors, strength of trunk extensors, and age. A stepwise selective multiple regression analysis for systolic pressure resulted in four significantly correlating variables: body fat percentage (p less than 0.001), heart rate (p less than 0.01), lean body mass (p less than 0.05), and strength of trunk extensors per kg body weight (p less than 0.05). For diastolic pressure the analysis resulted in two explaining variables: body mass index (p less than 0.001) and age (p less than 0.05). In a regression equation with 13 variables the strength of trunk flexors was negatively correlated with diastolic pressure. It is concluded that both fatness and muscularity are factors related to blood pressure in young men. The muscularity effect is more clearly associated with trunk and leg extensor strength.


The influence of exercise on muscle lysosomal enzymes. Representative lysosomal enzyme activities were measured in muscles taken 0 h and 24 h after an acute exercise run that was completed without any overt signs of fatigue. The animals had progressed 2 and 4 weeks into a standard exercise program which typically produces adaptive changes in the working muscles. There was an increase in acetylglucosaminidase activity (12%) in the fast-twitch red muscle section of all animals that participated in the training program. This small increase may be representative of a delayed response found after more exhausting exercise. The single exercise bout, however, did not cause any acute change in lysosomal activity nor alter the partition of lysosomal enzymes between the "free" and particulate fractions. Thus, altered lysosomal enzyme activity does not appear to be a contributing influence that challenges muscle fiber homeostasis during moderately intense running.


The effect of changes in cardiac frequency on left and right ventricular dP/dt max at different contractile states of the myocardium. In 17 canine heart-lung preparations the dependence of frequency potentiation of the right and left ventricular myocardium on the basic inotropic state of the heart was investigated. The effect of unipolar stimulation of the right atrium on dP/dt max in both ventricles was measured. The aortic pressure was maintained constant. Shortly after isolation of the heart, a stepwise increase of rate from 140 to 200 beats/min only had a very weak influence on left ventricular dP/dt max. With deterioration of the myocardium the frequency potentiation of dP/dt max increased considerably. End-diastolic pressure regularly decreased with rising cardiac frequency. Since the real positive inotropic effect is masked by the concomitant fall in diastolic loading, the end-diastolic pressure was maintained constant in a second group of 8 hearts during rate variation. The most pronounced inotropic effect was now found shortly after isolation of the heart. A rate increase of 30 beats/min resulted in a 20% rise of dP/dt max. The frequency potentiation decreased with deterioration of the heart resulting in a 12% dP/dt max increase at an estimated inotropic state of 50% of control. When the contractile state of the heart was improved above the control state by calcium application the frequency potentiation of the myocardium decreased. In the right ventricle similar results were obtained except for the fact that no significant correlation between the steepness of the frequency characteristics and the contractile state of the heart could be found when the end-diastolic pressure was kept constant.


Effect of long-term training and acute physical exercise on red cell 2,3-diphosphoglycerate. A statistically significant 10% increase (p less than 0.005) in mean red cell 2,3-diphosphoglycerate (2,3-DPG) concentration, concomitantly with a mean 16% increase (p less than 0.001) in the predicted maximal oxygen uptake (VO2max) was observed in 29 recruits, who were studied during 6 months of physical training in military service. The increase in 2,3-DPG was higher, the lower the initial 2,3-DPG and VO2max levels. The mean initial 2,3-DPG level was higher in the subjects with a higher initial VO2max. A strenuous but highly aerobic 21-km marching exercise elicited a mean 9% increase (p less than 0.005) in red cell 2,3-DPG concentration. A significantly greater response of 2,3-DPG to marching exercise was observed in subjects with a lower pre-test VO2max than in those with a higher pre-test VO2max. During another more competitive march 2,3-DPG remained almost unchanged and was associated with a tendency towards a negative correlation with the acccompanying lactate response (r = -0.60, p less than 0.05). Red cell 2,3-DPG response to a standardized exercise is considered to be a suitable indicator for evaluating the effect of training on an individual.


Purification and properties of hydrogenase from Megasphaera elsdenii. A hydrogenase has been purified to homogeneity from the soluble fraction of the rumen bacterium Megasphaera elsdenii, the overall purification is 200 times with a yield of 14%. The pure enzyme consists of a single polypeptide chain with Mr approximately 50 000 which contains 12 atoms of non-haem iron and 12 atoms of acid-labile sulphide. The enzyme is rapidly inactivated by O2 and it is therefore purified under nitrogen and in the presence of sodium dithionite. The optical spectrum of the enzyme, after removal of the dithionite with air, shows a peak at 275 nm (epsilon 275 nm = 143 mM-1 cm-1) and a shoulder between 350 nm and 400 nm (epsilon 400 nm = 46 mM-1 cm-1). The enzyme catalyses hydrogen production from sodium dithionite at a low rate. The rate is greatly enhanced by addition of the electron donors flavodoxin, ferredoxin and methyl viologen. The kinetic data with these three electron donors suggest co-operativity, but no indication of self-association of the enzyme was obtained. Sodium chloride enhances the rate of hydrogen production with methyl viologen semiquinone and changes the kinetic behaviour of the enzyme with this electron donor, but causes inhibition of the reactions mediated by ferredoxin and flavodoxin. Two kinetic models were developed which are consistent with the kinetic data of the three electron donors tested. The apparent co-operativity for the hydrogen production can be fitted with the mathematical form of those models. The identical kinetic behaviour of the hydrogenase with the one-electron donors flavodoxin and methyl viologen semiquinone monomer and the two-electron donor ferredoxin indicates that the hydrogenase accepts two electrons in two separate, independent steps and further indicates that the two (4Fe-4S) clusters of the donor ferredoxin are independent. The interpretation of the kinetic data with methyl viologen semiquinone is complicated by the fact that the semiquinone dimerises, and that the formation of the dimer is enhanced by salt. Taking into account the association of this donor, the activity of the enzyme with methyl viologen semiquinone can be described by the sum of the activities of the enzyme with methyl viologen monomer and methyl viologen dimer. The enzyme catalyses the oxidation of hydrogen gas with methyl and benzyl viologen as electron acceptors to their semiquinone forms; both electron acceptors show Michaelis-Menten kinetics. The hydrogen oxidation activity with both electron acceptors is stimulated by addition of sodium chloride. The kinetic data of the oxidation of hydrogen with the two-electron acceptors used are consistent with the porposed models, if it is assumed that the pathway followed is compulsory. At this moment no choice can be made between the models proposed.


1,4-alpha-Glucan phosphorylase from the slime mold Physarum polycephalum. Purification, physico-chemical and kinetic properties. Glycogen phosphorylase from macroplasmodia of Physarum polycephalum was purified 76-fold to homogeneity. The native enzyme migrated as a single protein band on analytical disc gel electrophoresis coinciding with phosphorylase activity. After reduction in the presence of sodium dodecylsulfate one protein band was detectable which corresponded to an Mr of 93 000. The molecular weight of the native enzyme determined by gel sieving or gradient-polyacrylamide gel electrophoresis was 172000 and 186000, respectively. The enzyme contained about 1 mol pyridoxal 5'-phosphate and less than 0.1 mol covalently bound phosphate per mol subunit. The amino acid composition of the enzyme was determined. In the direction of phosphorolysis the kinetic data were determined by initial velocity studies, assuming a rapid equilibrium random mechanism. Glucose 1-phosphate and GDP-glucose were competitive inhibitors toward phosphate and noncompetitive to glycogen. 5'-AMP, a weak activator of the enzyme, counteracted the glucose-1-phosphate inhibition completely. Physarum phosphorylase was compared with phosphorylases from other sources on the basis of chemical and kinetic properties. No evidence for the presence of phosphorylated forms has yet been found.


Inhibition of DNA synthesis in mammalian cells by daunomycin. Preferential inhibition of replicon initiation at low concentrations. The effect of the intercalating agent daunomycin on DNA synthesis was studied in cultured bovine liver cells. At low daunomycin concentrations (1 and 2 muM) the rate of [3H]thymidine incorporation decreased progressively with the duration of exposure to the inhibitor. This was accompanied by a shift of nascent DNA intermediates of replicon size to higher sedimentation values on sucrose gradients, indicating that daunomycin preferentially affects the initiation of replicating units, both in asynchronous and synchronized cells. At high daunomycin concentrations (12 muM) the rate of chain growth was also markedly reduced. This was indicated by a rapid and nearly complete cessation of the [3H]thymidine incorporation and an accumulation of nascent DNA intermediates of low molecular weight. These observations are discussed in relation to a pre-fork mode of DNA synthesis.


Androgenic regulation of messenger RNA sequence complexity in accessory sexual tissues of the male rat studied with fractionated complementary DNA. Effects of androgens on mRNA sequence complexity in the rat seminal vesicle have been investigated using complementary DNA fractionated on the basis of sequence abundance. Total cDNA complementary to poly(A)-rich RNA from normal rats was hybridised with an excess of the same RNA to controlled rot values and then the free cDNA was separated from cDNA . RNA hybrids by hydroxyapatite chromatography. Three cDNA fractions were obtained with very different hybridisation characteristics. Abundant cDNA hybridised to an excess of its parental RNA with an rot 1/2 of 2.46 x 10(-3) mol 1(-1) s and is complementary to about six or seven average-sized sequences. Use of hybrid-arrested translation in a cell-free protein-synthesising system has shown that this class of mRNA includes mRNAs coding for major androgen-dependent secretory proteins. Moderate and scarce cDNA fractions each showed more complex hybridization kinetics; computer analysis suggested each is complementary to two groups of average-sized sequences. Each cDNA fraction was hybridised to excess poly(A)-rich RNA from normal or castrated rats and the kinetics compared. Castration had no effect on the total number of sequences present in any class and did not alter the relative concentration of the scarce sequences. A small (threefold) decrease was seen in the concentration of abundant sequences with a larger (tenfold) decrease in the moderate class. Both de-reases were reversed by testosterone in vivo. The results are consistent with earlier studies where the effects of testosterone on seminal vesicle mRNA were followed using a translation assay and confirm that no gross differential effects are exerted on abundant mRNA coding for major secretory proteins. The cDNA fractions were also used to investigate the overlap in genetic expression between seminal vesicle and ventral prostate. Both tissues share all the scarce sequences in the same relative abundance. Less than 0.0015% and 0.004% of prostatic mRNA is complementary to seminal vesicle abundant and moderate sequences respectively. Similarly prostatic abundant sequences account for less than 0.004% of seminal vesicle mRNA.


Ligand competition curves as a diagnostic tool for delineating the nature of site-site interactions: theory. A few molecular models have been developed in recent years to explain the mechanism of cooperative ligand binding. The concerted model of Monod, Wyman and Changeux and the sequential model of Koshland, Némethy and Filmer were formulated to account for positively cooperative binding. The pre-existent asymmetry model and the sequential model can account for negatively cooperative ligand binding. In most cases, however, it is virtually impossible to deduce the molecular mechanism of ligand binding solely from the shape of the binding isotherm. In the present study we suggest a new strategy for delineating the molecular mechanism responsible for cooperative ligand binding from binding isotherms. In this approach one examines the effect of one ligand on the cooperativity observed in the binding of another ligand, where the two ligands compete for the same set of binding sites. It is demonstrated that the cooperativity of ligand binding can be modulated when a competitive ligand is present in the protein-ligand binding mixture. A general mathematical formulation of this modulation is presented in thermodynamic terms, using model-independent parameters. The relation between the Hill coefficient at 50% ligand saturation with respect to ligand X in the absence, h(x), and in the presence of a competing ligand Z, h(x,z), is expressed in terms of the thermodynamic parameters characterizing the binding of the two ligands. Then the relationship between h(x) and h(x,z), in terms of the molecular parameters of the different allosteric models, is explored. This analysis reveals that the different allosteric models predict different relationships between h(x,z) and h(x). These differences are especially focused when Z binds non-cooperatively. Thus, it becomes possible, on the basis of ligand binding experiments alone, to decide which of the allosteric models best fits a set of experimental data.


HeLa cell cytoplasmic mRNA contains three classes of sequences: predominantly poly(A)-free, predominantly poly(A)-containing and bimorphic. The mRNA species which exist in the HeLa cell polyribisomes in a form devoid of A sequences longer than 8 nucleotides constitute the poly(A)-free class of mRNA. The rapidly labelled component of this mRNA class shares no measurable sequence homology with poly(A)-containing RNA. If poly(A)-free mRNA larger than 12 S labelled for 2 h in vivo is hybridized with total cellular DNA, it hybridizes primarily with single-copy DNA. When a large excess of steady poly(A)-containing RNA is added before hybridization of labelled poly(A)-free RNA, no inhibition of hybridization occurs. This indicates the existence of a class of poly(A)-free mRNA with no poly(A)-containing counterpart. Some mRNA species can exist solely as poly(A)-containing mRNAs. These mRNAs in HeLa cells are found almost exclusively in the mRNA species present only a few times per cell (scarce sequences). Some mRNA species can exist in two forms, poly(A)containing and lacking, as evidenced by the translation data in vitro of Kaufmann et al. [Proc. Natl Acad. Sci. U.S.A. 74, 4801--4805 (1977)]. In addition, if cDNA to total poly(A)-containing mRNA is fractionated into abundant and scarce classes, 47% of the scarce class cDNA can be readily hybridized with poly(A)-free mRNA. 10% of the abundant cDNA to poly(A)-containing mRNA will hybridize with poly(A)-free sequences very rapidly while the other 90% hybridize 160 times more slowly, indicating two very different frequency distributions. The cytoplasmic metabolism of these three distinct mRNA classes is discussed.


The binding of 8-anilinonaphthalene-1-sulphonate to a fixed concentration of unenergised and succinate-energised submitochondrial particles. The effect of the approximately hyperbolic relationship between fluorochrome concentration and light absorbed on the interpretation of data for the binding of 8-anilinonaphthalene-1-sulphonate to unenergised and succinate-energised submitochondrial particles has been investigated. If this is taken into account plots of fluorescence against fluorescence x [fluorochrome]-1 do not tend towards a maximum fluorescence value. The significance of this findings is discussed.


Evidence by chemical modification for the involvement of one or more tryptophanyl residues of bovine antithrombin in the binding of high-affinity heparin. Tryptophanyl residues of bovine antithrombin were modified with N-bromosuccinimide at near-neutral pH. The reaction was found to be specific for tryptophan at low levels of modification, i.e. when only up to 1--1.3 mol tryptophan/mol protein were oxidized. Further modification led to extensive side reactions. Modification of an average of about one tryptophanyl residue per protein molecule did not affect antithrombin activity measured in the absence of heparin, but decreased the activity assayed in the presence of heparin to about half the value given by unmodified antithrombin. Addition of an excess of high-affinity heparin to a similarly modified antithrombin sample resulted in much smaller circular dichroism, ultraviolet absorption and fluorescence changes than those observed with the intact protein. Modification experiments in the presence of excess high-affinity heparin gave a definitely lower extent of modification than when heparin was excluded. These studies thus reinforce the conclusion from previous spectroscopic analyses that one or more tryptophanyl residues of antithrombin are involved in the binding of high-affinity heparin, presumably by being located at or close to the heparin binding site.


Transport of D-lactate in perfused rat liver. The transport of D-lactate across the plasma membrane was investigated in hemoglobin-free perfused rat livers, applying the multiple-indicator dilution technique (pulse labelling of D-lactate and indicator substances). The following results were obtained: 1. The steady state exchange rate at 1 mM D-lactate was 2.5 mumol x min-1 x g wet wt-1. It was proportional to the extracellular concentration in the range between 0.1 and 70 mM. 2. The transport of D-lactate was inhibited by L-lactate and pyruvate; 50% inhibition was observed at 40 mM L-lactate or 5 mM pyruvate. 3. The transport was also inhibited by alpha-cyanocinnamate and 4,4'-diisocyanostilbene-2,2'-disulfonic acid. The inhibition by cyanocinnamate was complete (with 25 mM) and fully reversible, whereas the inhibition by diisothiocyanostilbenedisulfonic acid was incomplete and irreversible; it was dependent upon the amount of diisothiocyanostilbenedisulfonic acid bound by the liver. Maximal inhibition (80%) was observed with 2 mumol diisothiocyanostilbenedisulfonic acid bound per g wet weight. 4. The intracellular concentration (ci) of D-lactate was proportional to the extracellular concentration (ce); the ratio ci/ce was 0.5 throughout the concentration range studied. It decreased in the presence of L-lactate or pyruvate. It is concluded that the transport of D-lactate is carrier-mediated, and, at least partially, electroneutral.


Structural characterization by nuclear magnetic resonance of a reactive-site 13carbon-labelled basic pancreatic trypsin inhibitor with the peptide bond Arg-39--Ala-40 cleaved and Arg-39 removed. With the use of an enzymatic replacement method, 90%-enriched [1-13C]lysine was introduced into the reactive site of the basic pancreatic trypsin inhibitor. Characterization of the labelled inhibitor with 13C nuclear magnetic resonance (NMR), 1H NMR and chemical methods showed that while the reactive-site peptide bond Lys-15--Ala-16 was properly resynthesized, the polypeptide chain was cleaved at the peptide bond Arg-39--Ala-40 and Arg-39 was removed. Detailed 1H NMR studies showed further that, with the exception of the immediate environment of the modification site, the average spatial structure of the native inhibitor was preserved in the modified protein. Compared to the native inhibitor, the thermal stability of the globular conformation was found to be reduced, interior amide protons exchanged at a faster rate and the internal mobility of aromatic rings located outside the immediate environment of the cleaved peptide bond was essentially unchanged. These observations coincide closely with previous reports on different modifications of the inhibitor and can be explained by a recently proposed dynamic multi-state model for globular proteins. Since the fundamental structural properties of the native inhibitor and full inhibitory activity are preserved after resynthesis, the [1-13C]lys-15-labelled inhibitor with the peptide bond Arg-39--Ala-40 cleaved and Arg-39 removed should be suitable for 13C NMR studies of mechanistic aspects of proteinase-inhibitor interactions.


Specific labeling of a phosphate-transporting protein from rat-liver mitochondria by [203Hg]mersalyl. 1. A highly specific labeling of a phosphate-transporting protein from rat-liver mitochondria was obtained with [203Hg]mersalyl under the following conditions: (a) labeling of mitoplasts, isolated by the French-Press procedure; (b) washing of the isolated inner membranes with NaCl/octylglucoside. 2. The concentration-dependent labeling of the protein with mersalyl showed a biphasic saturation which correlated well with the protection and inhibition of phosphate transport. 3. The molecular weight (Mr 31000) and the amount of labeled carrier protein (24 nmol/g mitochondrial protein) at 100% protection correlated well with published data from the literature. 4. Gel filtration of the labeled protein in the presence of non-ionic detergents showed a molecular weight of at least twice of that obtained by gel electrophoresis in the presence of sodium dodecylsulfate.


Pharmacokinetics of fenfluramine enantiomers in man. The pharmacokinetics of fenfluramine enantiomers were studied in normal volunteers following the administration of single of multiple doses of racemic fenfluramine hydrochloride. Plasma concentrations and half-lives were similar for both enantiomers after single 40 mg of 60 mg doses. However following chronic administration (2 x 40 mg for 10 days), significant differences were observed between the kinetic parameters of the two enantiomers, the l-form of fenfluramine and norfenfluramine accumulating in plasms more than the d-forms.


The fate of ticlopidine in the organism. II.--Distribution and elimination of ticlopidine 14C after a single oral administration in the rat. In the Rat, distribution and elimination of Ticlopidine C14 after a single oral dose of 25 mg/kg were studied and compared with that observed after a single intravenous injection of the same dose. The plasma decreases of total radioactivity were found to be identical regardless of adminstration route, indicating almost complete absorption of the labelled molecule. The elimination half-life, which was 1.56 +/- 0.08 days, was equivalent to the half-life measured after intravenous injection, and no difference in tissues distribution was observed. As in the case of intravenous injection, the radioactivity was eliminated principally in the feces and only to a slight degree in the urine.


Quantitative determination of praziquantel in body fluids by gas liquid chromatography. Praziquantel was determined in body fluids by gas liquid chromatography as follows: A known amount of an internal standard and 0.1 N sodium hydroxide solution was added to the sample to be analyzed. After extraction with methyl acetate/diisopropyl ether = 30/70, the organic extract was evaporated, the residue taken up in methylacetate and an aliquot injected for glc analysis. Separation was accomplished on a OV3 silicon oil phase, and for detection and quantitation, a thermoionic FID sensitized for nitrogen-containing compounds was used. The determination limit in serum is about 0.01 micrograms/ml. The relative standard deviation for serum concentrations of 0.1 micrograms/ml was found to be 4.5%.


A fluorometric method for the determination of praziquantel in blood-plasma and urine. Some physicochemical data of praziquantel which may have analytical relevance are reported. For the quantitative determination praziquantel is extracted from plasma or urine by means of organic solvents and then hydrolyzed in an aqueous alkaline solution. The hydrolyzed product is reacted with dansyl-chloride (5-dimethylaminonaphthalene-sulfonylchloride). The dansylated compound is separated and quantified fluorometrically. The limit of determination is 3 micrograms/l in blood plasma and 30 micrograms/d in urine. For both fluids, the imprecision is approximately 7.5%. The method is suitable for the determination of praziquantel in patients or healthy volunteers treated with therapeutic doses.


Localization of S-[methyl-14 C]adenosyl-L-methionine in pregnant mice and fetuses as determined by autoradiography. The distribution of S-[methyl-14C]Adenosyl-L-methionine has been examined in pregnant mice using an autoradiographic technique. The results indicate that the compound crosses the placental barrier quite slowly and accumulates in some fetal tissues such as intestine, liver, kidney, eye and lung. The highest concentration is reached 12 hours after i.v. administration of the compound. In the placenta the labelling can be detected for a long time after administering the radioactive compound.


Esterase activity of rat muscle.

Esterase activity of rat muscle. The esterasic capacity of a series of skeletal muscles in response to three hemisuccinate ester drugs was investigated in rats and compared to that on alpha-naphthylacetate as a reference esterase substrate. Marked variations between different muscles and between given muscles of animals of different sex were observed, indicative of a complex heterogeneity in muscular expression of esterase activity.


A correlation of clinical findings and CT in ischaemic cerebrovascular disease. 248 patients with an ischaemic stroke were investigated with computer tomography. A hypodense lesion was observed in 18% of TIAs, 76% of PRINDS and 95% of completed stroke patients. In the patients with TIA the hypodense lesions were mainly single ones, whereas completed stroke patients also had a significantly higher proportion of more bilateral lesions. Atrophy was present in 44% of TIA patients, 68% of PRIND patients and 82% of completed stroke patients. Atrophy was generalised in 60% of the patients and homolateral in only 16%. Contrast medium enhancement in a hypodense area occurred only in patients with PRIND and completed stroke and never in TIA aptients. Multiple lesions and atrophy were significantly more common in patients with dementia. The time course showed oedema initially in 47% of patients which fell to 25% after the 1st week. Contrast medium enhancement was present initially in 42% of the patients, rose to 70% until the 3rd week and then fell continuously until the 6th week. The change from an indistinct to a distinct outline of a lesion coincided with this.


Hematin and propranolol in acute intermittent porphyria. Full recovery from quadriplegic coma and respiratory failure. The authors present a case of acute intermittent porphyria (AIP) in an almost fatal relapse with quadriplegia, bulbar paralysis and coma. Intravenous hematin produced an immediate arousal from coma and allowed a gradual resumption of bulbar and autonomic functions. Persistent tachycardia and hypertension necessitated huge doses of intravenous propranolol. Both hematin and propranolol administrations were followed by a remarkable decrease in urinary amino-levulinic acid and porphobilinogen excretion. Nevertheless, after the acute stage, the patient was left with a severe generalized muscle wasting. After 7 months of intensive physical therapy, complete recovery of all neuromuscular functions was achieved. The modern aspects of the management of AIP are presented; the efficacy and the limits of hematin and propranolol therapy are discussed.


The autoregulation of cerebral blood flow, the cerebrovascular reactivity and their interaction in the Shy-Drager syndrome. The experimental study of the autoregulation of cerebral blood flow (CBF), of the cerebrovascular reactivity and of their interaction is performed in a confirmed case of Shy-Drager syndrome, by means of the xenon-clearance technique. The basal mean regional CBF is slightly depressed. The cerebrovascular reactivities to variation in paCO2 and to local cerebral activation are maintained. The inferior threshold of breaking of autoregulation is raised. A competition between the effects on CBF of autoregulation loss and of metabolic regulation is demonstrated. The latency of operation of the autoregulation is evoked by the analysis of the CBF measurements and is confirmed by the simultaneous EEG study.


Infantile form of so-called neuronal ceroid lipofuscinosis: lipid biochemical studies, fatty acid analysis of cerebroside sulfatides and sphingomyelin, myelin density profile and lipid composition. The biochemical analysis of a case of infantile neuronal ceroid lipofuscinosis, as determined by clinical and neuropathological findings, is presented. A diminished amount of solids is found, the amount of lipids is 30% of the normal as expressed in lyophilized tissue. The yield of myelin isolated by the density gradient is 1.8% of the normal. Phospholipid patterns show a reduction in ethanolamine phosphoglyceride, N-acetylneuraminic acid is extremely low and sphingolipids are largely reduced, cerebrosides being most affected (2.5% of the normal). In cerebrosides and sulfatides the decrease in very long chain fatty acids is important, but the deficiency in any type (including hydroxy compounds) is not too dramatic. According to the aspect under electron microscopy, the density profile, and the biochemical composition of the subfractions, isolated myelin is close to normal. The loss of the myelin sheath appears to reflect a Wallerian degeneration in the CNS: myelin loss is a secondary effect. This disease, from a biochemical point of view, seems to be the ideal control for leukodystrophies.


Immunoglobulin G in serum and cerebrospinal fluid from epileptic patients treated with phenytoin. Concentrations of IgG and titres of IgG antibodies to rabbit erythrocytes were studied in serum and cerebrospinal fluid (CSF) from 21 phenytoin-treated patients; 11 of these had reduced serum IgA. The CSF IgG concentration was not reduced in any patient. A reduction of CSF IgG has been found earlier in phenytoin-treated patients, when other antisera to IgG were used for the determination of IgG concentration. The present findings therefore indicate that an alteration of the composition or quality of the CSF IgG is present in phenytoin-treated patients. 1 patient had a phenytoin-induced low serum concentration of IgG and IgA. In this patient the ratios CSF:serum were normal both for IgG concentration and for IgG antibodies to rabbit erythrocytes.


The sink action of cerebrospinal fluid in uremia. The distribution of water, electrolytes and non-electrolytes in various compartments of the central nervous system was analyzed in rats subjected to various stages of uremia. After bilateral nephrectomy for various periods of time ranging from 1 to 18 h, there is a progressive hyperkalemia, hyponatremia and metabolic acidosis. Although the cerebrospinal fluid (CSF) [K] progressively increases to a concentration of 6 mmol/l at 48 h post-nephrectomy, the ratio of CSF [K]/plasma [K] remains relatively constant throughout the duration of the uremic challenge; on the other hand, the concentration gradient for Na (CSF to plasma) markedly increases with progression of uremia. The steady rise in cellular [K] in both the choroid plexus and in at least one compartment of the cerebral cortex is presumably a reflection of stimulattion of the Na-K pump caused by a build-up in [K] in the extracellular fluids of the brain. The blood-brain and blood-CSF barrier systems to both 14C-urea and 131I remain intact 24--48 h after nephrectomy; and there is not any evidence of cerebral edema at any stage of uremia. Thus, several lines of evidence indicate that the CSF-sink effect is maintained even in advanced uremia.


Hyperparathyroidism presenting with unusual neurological features. A case of hyperparathyroidism, who presented with unusual complex neurological symptomatology is reported. Mental confusion, asterixis and elevated cerebrospinal fluid protein with marked pleocytosis dominated the neurological picture. All these findings subsided upon correction of hypercalcemia, suggesting direct relationship between cerebrospinal fluid abnormalities, asterixis and this metabolic disturbance.


Effectiveness of simplified dosage schedules on the management of ambulant epileptic patients. The effect of dosage schedules on plasma drug levels was evaluated in two groups of ambulant epileptic patients, whose treatment with diphenylhydantoin (DPH) or phenobarbital (PB) had been simplified by reducing the daily frequency of the dosage regimens. Although the patients received the same milligram per kilogram daily dosages of DPH or PB during the study, the mean plasma levels of the two drugs were significantly higher during the less-frequent dosage regimens. The results obtained suggest that twice-daily dosage schedules of DPH and once-daily dosage schedules of PB are advantageous over more complicated dosage schedules resulting in a higher compliance among ambulant epileptic patients.


Porto-hepatic bypass by organohepatopexy in rats with prehepatic portal hypertension. Formation of new venous porto-hepatic collaterals following omento-, jejuno-, and splenohepatopexy, and their effect on portal vein pressure. Transposition to the liver of the greater omentum; the jejunum, and the spleen in 3 groups of rats with prehepatic portal hypertension due to a calibrated portal vein stenosis with a diameter of 1.2 mm resulted in the formation of new venous collaterals from the transposed organs into the liver parenchyma. The collaterals were visualized macro- and microscopically, as well as angiographically, and were found to drain into intrahepatic branches of the portal vein, sinusoids, and hepatic veins. The portal vein pressure 8 weeks after stenosis and transposition was found to be significantly lower in rats with transposed organs than in controls. The collaterals were sufficiently well developed to significantly reduce the increase in portal vein pressure that follows acute occlusion of the portal vein and the natural splenorenal collaterals.


Amino acid incorporation into liver proteins during hemorrhagic shock in the dog. Liver protein synthesis was studied with an in vitro method during and after 1 h of hemorrhagic shock in the dog. Protein synthesis was significantly decreased already 15 min after induction of hypovolemia and was about 50% of the initial value at the end of the shock period. 2 h following retransfusion, protein synthesis was normalized. Some of the underlying mechanisms leading to decreased protein synthesis in hemorrhagic shock are discussed. Serum concentrations of liver enzymes were unchanged during the shock period indicating that the liver metabolism may be affected without changes in parameters often used to evaluated liver function.


Gastrointestinal propulsion and mixing in the rat during infusion of dopamine. The influence of continuous intravenous infusion of dopamine on gastric evacuation, small bowel propulsion and mixing was studied in the rat. The study revealed no influence of dopamine infusion on gastrointestinal transport mechanisms.


The effect of dextran on the platelet distribution and lysability of ex vivo thrombi in dogs. Dextran infusion causes an increased lysability of ex vivo thrombi. This increased lysability is shown to be accompanied by a change in the distribution of 51Cr-labelled platelets. Rather than being localized to the head of the thrombus, after dextran infusion the platelets are found to be more evenly distributed. This change in platelet distribution is secondary to an effect of dextran on platelet function, probably via previously demonstrated impairment of the function of factor VIII. It is suggested that the increase in thrombus lysability is due to the change in platelet function.


Infusion of purified antithrombin III to patients with fracture of the neck of the femur. A purified antithrombin III (AT) preparation was infused in 8 patients with hip fractures, while 8 patients were observed as controls. The infusion was well tolerated by all patients, and the AT activity in plasma was significantly raised throughout an observation period of 48 h as compared with the controls.


Circulating tissue thromboplastin during hip surgery. An immunoradiometric assay for tissue thromboplastin has been established. Blood levels in 6 patients undergoing total hip replacement have been determined. In 3 patients, high levels of circulating apoprotein III were found at various stages of the operation, showing a release of tissue thromboplastin chiefly during impaction of the prosthesis into the femoral bone. The other 3 patients had low or undetectable levels.


Age-associated changes in activities of rat hepatocytes. I. Protein synthesis. Experiments involving simultaneous comparisons of cells from two animals establish an age-dependent decline in protein synthesis. Specific radioactivities of fractions from cells of aged animals (20+ months) are depressed 60% to 80% versus those from younger animals (2 to 14 months). Isotope ratio patterns derived by PAGE indicate that formation of a 20000 MW polypeptide of cytoribosomal origin is even more strongly and specifically depressed.


Ventilation, lung volumes and lung mechanics of young adult and old Syrian hamsters. The breathing pattern, subdivisions of lung volume and quasistatic pressure-volume relationships of 15 and 65 week old hamsters (Mesocricetus auratus) anesthetized with halothane were measured by plethysmography. Body weights, tidal volumes and minute volumes of the younger and older groups were nearly identical. All lung volumes were larger in the older hamsters but there were no significant age-related differences in the relative proportions of lung volume subdivisions. Quasistatic lung compliance was greater in the older group but differences in pressure-volume relationships were not significant when volume was expressed as percent of total lung capacity. It appears that hamster lungs undergo significant volume changes with age during adulthood, but the pattern of change is different from that observed in man and dogs.


Life span and pathology in offspring following nicotine and methamphetamine exposure. Seventy-nine primiparous Sprague-Dawley rats received s.c. injections of either 3.0 mg/Kg of pure nicotine, 5.0 mg/Kg of methamphetamine HCL, 5.0 MG/Kg of saline vehicle, or received no injections during the 21 day gestational and 19 day nursing period. Male offspring were divided into 3 groups at weaning. The autopsy animals (n = 80) were sacrificed at selected periods during the life span, the aging animals (n = 60) were weighed monthly until death, and the behavioral rats (n = 48) were monitored on a variety of behavioral tasks throughout their life span. Measures reported here include life span data, organ pathology and tumor formation. Significant differences among the four offspring groups were found for tumor incidence, gross pathology, and age at death in the autopsy group; and weight maintenance in the aging group.


Living arrangements and well being in elderly women. Psychosocial and health data were obtained from 50 black and 47 white females age 65 and over. The women were divided according to whether they lived alone, with spouse only, or with persons other than (or in addition to) the spouse. Those living alone were found to have lower life satisfaction, lower self-esteem, and higher social dysfunction than those living with spouse or others. No differences were found with respect to depression, somatization, diet, or activity level. There was a significant interaction effect between race and living arrangement with respect to anxiety, with blacks being less anxious than whites when living with others, and more anxious when living alone or with only a spouse.


Tumor induced brain damage in rats: implications for behavioral and anatomical studies with aging animals. In rats, spontaneously occurring tumors and infarcts induce damage which destroys or distorts regions of the brainstem and cerebral hemispheres. Despite such destruction, there may be no gross evidence of behavioral or metabolic pathology.


Age differences in the consistency of adults' associative responses. Younger (mean age = 20) and older (mean age = 63) adults were asked to produce free associations to the same list of 24 verbal stimuli on each of four trials. Older adults tended to produce less common associations and their associations were less consistent across trials. Possible relationships between older adults' inconsistency in processing and their (a) poorer memory performance and (b) slower reaction times were discussed.


Comparison of the antitussive effects of codeine phosphate 20 mg, dextromethorphan 30 mg and noscapine 30 mg using citric acid-induced cough in normal subjects. 1. Protection by codeine 20 mg, dextromethorphan 30 mg, noscapine 30 mg, and placebo against citric acid-induced cough was determined in eighteen healthy subjects. 2. Drug differences occurred at 2 1/2 h following ingestion of the drugs but not at 1 1/4 h. 3. Only coideine 20 mg had a greater antitussive action than placebo, but dextromethorphan 30 mg also did not differ from codeine 20 mg. 4. This technique may offer a useful screening test for the activity of new potential antitussive compounds in man.


The pharmacokinetics of diclofenac sodium following intravenous and oral administration. The pharmacokinetics of diclofenac were examined following single rapid intravenous injection and also following single oral doses to healthy female volunteers. After intravenous injection plasma levels of diclofenac fell rapidly and were below the limits of detection at 5.5 h postdosing. Individual drug profiles were described by a triexponential function and mean half-lives of the three exponential phases were 0.05, 0.26 and 1.1 h. After oral doses of enteric-coated tablets, the lag time between dosing and the appearance of drug in plasma varied between 1.0 and 4.5 h. However once drug absorption had commenced similar plasma drug profiles were obtained in different individuals. Peak plasma diclofenac levels ranged from 1.4 to 3.0 microgram . ml-1. The mean terminal drug half-life in plasma was 1.8 h after oral doses. This value was not significantly greater than the value of 1.1 h following intravenous doses. Fifty percent of orally dosed diclofenac did not reach the systemic circulation due, predominantly, to first-pass metabolism.


The disposition of intravenous doxapram in man. The pharmacokinetics of intravenous doxapram in healthy individuals is consistent with a three-compartment open model. Doxapram was administered by bolus injection (1.5 mg . kg-1) and by intravenous infusion (6.5 mg . kg-1 for 2 h) to 5 subjects on separate occasions. There was no significant difference in mean terminal plasma half-lives (355 and 448 min) or in mean total body clearances 5.9 and 5.6 ml . min-1 . kg-1) following i.v. bolus injection or infusion respectively. In 3 subjects plasma doxapram concentrations during and after i. v. infusion agreed with those predicted from pharmacokinetic values obtained from the bolus injection study. Since mean steady-state concentrations (9.9 microgram . ml-1) would be reached only after an extended interval (mean 15.2 h), a variable-rate infusion regimen was calculated to produce and maintain a concentration of 2 microgram . ml-1 from 15--25 min onwards. A regimen in which the infusion rate is reduced step-wise is recommended to achieve early near-constant plasma doxapram concentrations.


Ergotism in a patient with a portosystemic shunt. A case of ergotism is described in a patient with Banti's syndrome in whom a porto-systemic shunt had been made three times. She was admitted to hospital with severe ischaemia of the extremities after having taken a single oral dose of 1 mg ergotamine tartrate. Her condition improved rapidly after 2 to 3 days and the circulation was normal after one week. The possible mechanism of ergotism in this patient is discussed. Although the plasma concentration of ergotamine was not measured, the likeliest explanation was that the serious side effects were related to the altered pharmacokinetics and loss of first-pass metabolism of the drug due to the venous bypass of the liver.


Differential effects of d- and l-propranolol on dopamine turnover stimulated by oxotremorine in striatal and mesolimbic areas of rat brain. The effects of l-propranolol, d-propranolol and clonidine on homovanillic acid (HVA) concentrations in the corpus striatum and nucleus accumbens of rats were studied under normal conditions and after treatment with oxotremorine or haloperidol. Thile propranolol and clonidine given alone had no significant effect on HVA levels in either area, l-propranolol (1--10 mg/kg) and clonidine (0.1 mg/kg), both significantly inhibited the elevation of striatal HVA, found 60 min after oxotremorine administration. Both l- and d-propranolol (2.5 mg/kg), reduced the effect of oxotremorine in the nucleus accumbens. Neither l-propranolol no clonidine affected the rise in HVA in either brain area seen after haloperidol. Our results suggest that propranolol may reduce cholinergic activation of dopaminergic pathways by two different mechanisms. One is stereospecific for the l-isomer and operates in the striatum and another, which is shared by both isomers, occurs in the nucleus accumbens.


Blockade of serotonin receptors on autonomic neurones by (-)-cocaine and some related compounds. The interaction between (--)-cocaine and responses to 5-HT elicited through serotonin receptors on autonomic neurones has been investigated on the rabbit heart and the guinea-pig ileum. Low concentrations of (--)-cocaine or its stereoisomer, (4)-pseudococaine, produced shifts to the right of the 5-HT dose-response curves on heart and ileum with no depression of the maximum responses to electrical stimulation or dimethylphenylpiperazinium remained unaffected. A Schild analysis of data obtained on heart and ileum indicated competitive antagonism of 5-HT by (--)-cocaine. Antagonism of 5-HT by the cocaine isomers cannot be ascribed to local anaesthesia per se since neither lignocaine, tetracaine, benzocaine nor bu tacaine were selective antagonists of 5-HT. Similarly, inhibition of monoamine uptake seems of minimal relevance since desipramine proved only a weak antagonist of 5-HT on the heart and did not influence the 5-HT antagonist potency of (--)-cocaine. Selective blockade of 5-HT neuronal responses is a property shared by several structural analogues of (--)-cocaine and (+)-pseudococaine; nor-(--)-cocaine proved the most potent of these, being active at a concentration of 2 x 10(-8) M. These data indicate that (--)-cocaine and several of its derivatives inhibit 5-HT stimulation of both adrenergic and cholinergic autonomic neurones through competition with the agonist at serotonin receptor sties. Since morphine, the tool normally used to identify responses mediated through neuronal serotonin receptors, acts only at certain "morphine-sensitive" junctions and then, non-discriminately, the cocaine analogues, and particularly nor-(--)-cocaine would seem to offer real advantages as tools for differentiating such responses.


Paradoxical GABA excitation of nigral dopaminergic cells: indirect mediation through reticulata inhibitory neurons. Biochemical and electrophysiological and electrophysiological studies suggest that GABA agonists increase the activity of dopaminergic neurons in the zona compacta (ZC) of the substantia nigra despite a known GABAergic input to ZC cells. Using single-unit recording techniques we have investigated this "paradoxical" effect. One population of neurons located in the zona reticulata (ZR) of the substantia nigra was found to be 20 times more sensitive to iontophoretically applied GABA than ZC neurons. GABA introduced by means of microiontophoresis into the ZR caused an increase in ZC cell activity while glutamic acid introduced in the same manner produced an inhibition of ZC cells. The latter effect was blocked by low doses of picrotoxin. Muscimol (i.v.) caused a decrease in ZR cell activity at the same dose that caused a parallel increase in ZC cell firing rate. These data suggest that ZC cells receive an inhibitory GABAergic input from ZR cells that are in turn inhibited by low doses of GABA agonists. Such an anatomical arrangement would account for the "paradoxical" excitatory effect of GABA agonists on ZC neuron activity.


Biphasic effects of morphine on cardiovascular system of the cat. Morphine has been reported to both lower blood pressure (BP) and cause excitation in some species. These are seemingly contradictory effects. In this series of experiments, the actions of morphine on BP, heart rate (HR) and pupillary diameter were re-examined in sedated, paralyzed cats under 2 ranges of dosage and 3 types of sedative. Small doses (1--2 mg/kg) lowered BP and HR while larger doses (8--12 mg/kg) increased both BP and HR. The effect of morphine on pupillary diameter, however, was not biphasic in that increasing doses proportionally increased pupillary diameter. In adrenalectomized cats smaller doses of morphine decreased, but larger doses no longer increased, BP and HR as it has in the non-adrenalectomized animals. Morphine also no longer produced mydriasis in the adrenalectomized cats. The results suggest that, in the cat, morphine can have a biphasic effect and that the adrenals may play a role in the excitatory phase.


Vagal bradycardia produced by microinjections of morphine-like drugs into the nucleus ambiguus in anaesthetized dogs. The site in the dog medulla oblongata where fentanyl or met-enkephalinamide produced vagal bradycardia was determined using microinjections. The nucleus ambiguus was found to be a selective and highly sensitive area for the vagal bradycardia.


Antidepressants selectively antagonize the hyperactivity induced in rats by long-term isolation. Rats isolated at 16--18 days of age showed after 10--12 months a complex behavioural syndrome in which locomotor activity was markedly increased when compared to that of group-housed controls. The hyperactivity of the socially deprived animals was selectively blocked by clinically effective antidepressants but not by other classes of psychotropic drugs. The present animal model may be of value for the detection of new antidepressants and contribute to elucidate the aetiology of depression.


Morphine suppression of substantia nigra zona reticulata neurons in the rat: implicated role for a novel striatonigral feedback mechanism. In lightly anesthetized rats, systemic administration of morphine exerted a naloxone-reversible suppression of spontaneously discharging neurons located primarily in the substantia nigra zona reticulata, either at rest or after their activation by microinjection of dopamine into the caudate nucleus. Inactivation of these neurons within the striatonigral feedback pathway is suggested to complement a direct action on the nigrostriatal dopaminergic cells by the opiate, resulting in the morphine suppression of caudate neuronal activities.


Effects of some antiarrhythmic agents on dog heart mitochondrial oxidative phosphorylation. Significant decreases in state 3 oxygen consumption, respiratory control index (RCI), ADP : O ratio and phosphorylation rate were observed in the presence of 0.1-1.0 mM quinidine; 0.5 mM or greater concentrations elevated state 4 oxygen consumption. Lidocaine (1-2.5 mM) also depressed RCI and increased state 4 oxygen consumption but procaine amide (1-2.5 mM) was without effect. It is suggested that cardiodepression by quinidine may be associated with its ability to impair mitochondrial metabolism.


Dopaminergic agonists and their influence on the oxygenation and function activity of underperfused brain tissue. The effect of two dopaminergic agonists (apomorphine and bromocriptine) on electrical activity and oxygen supply of the brain was investigated in cats submitted to hypovolemic oligemia (mean arterial blood pressure: 45 mmHg). While both drugs stimulated the brain by prolonging the oligemia-induced seizures in the caudate nucleus and in the cerebral cortex, only apomorphine improved the pO2 distribution in the cortical tissue after 120 min oligemia. Bromocriptine, in contrast, had a beneficial effect of shorter duration. These data show that under conditions of incomplete ischemia the brain can still be activated. Furthermore, these results provide additional support for the biochemically founded hypothesis of different dopamine receptors in the brain.


Yohimbine blocks lateral hypothalamus-mediated behaviors. There exist behavioral and physiological similarities between rats injected with yohimbine and those with lesions of the lateral hypothalamus. The present study sought to determine if behaviors mediated by the lateral hypothalamus would be significantly altered by yohimbine. Deficits in performance of an active avoidance task have been observed after lateral hypothalamic lesions. Likewise, in the present experiment, yohimbine significantly reduced the ability of trained rats to perform in a similar paradigm. Systemic injections of yohimbine caused decrements in lateral hypothalamic self-stimulation behaviors as well as in locomotor activity produced by lateral hypothalamic stimulation. Similar behaviors mediated by the substantia nigra were relatively unaltered by yohimbine. A final experiment revealed that yohimbine reduces the enhanced glucose utilization normally observed at the lateral hypothalamus during electrical stimulation. These data support the view that the lateral hypothalamus may be an important target of yohimbine's CNS actions.


Differential effects of acute and subacute HA-966 treatment on storage and release of striatal dopamine. Acute injections of HA-966 (100 mg/kg) into rats caused a rapid elevation of dopamine (DA) content in the striatum. 3,4-Dihydroxyphenylacetic acid (DOPAC) and homovanillic acid (HVA) levels increased after a latency period of 0.5 h and 1 h respectively. Repeated ("subacute") HA-966 treatment produced a smaller DA increase than did single administration, while DOPAC and HVA rose at the same rate after both treatments. In HA-966-pretreated rats no tolerance for DA increase followed a lesion of the dopaminergic fibres. Acute as well as subacute HA-966 prevented the disappearance of DA after alpha-methyl-p-tyrosine for about 3 h. In both cases DOPAC and HVA levels dropped sharply after HA-966. HA-966 had no influence on the decline of DOPAC and HVA levels after monoamine oxidase inhibition. It is concluded that the rises of DOPAC and HVA after HA-966 did not occur because the capacity of the vesicular DA stores was exceeded. Instead, HA-966 affects the storage mechanism for newly formed DA. Possible explanations for the observed tolerance to DA accumulation after HA-966 are discussed.


Sodium ions and the development of the inotropic action of ouabain in guinea-pig heart. Factors that determine the interaction of ouabain with the positive inotropic receptor were examined in isolated perfused guinea-pig hearts. The hearts were exposed to ouabain during a perfusion with a modified Krebs-Henseleit solution. The interaction of ouabain with the inotropic receptors under those conditions was estimated by subsequently perfusing the heart with a regular Krebs-Henseleit solution and monitoring the resting and developed tension. Ouabain failed to cause an increase in the force of contraction when the cardiac tissue was exposed to this agent in the absence of Na+ and Ca2+ either in the presence of a low or high concentration of K+. The absence of Ca2+ or the lack of contraction was not responsible for the failure of ouabain to interact with the inotropic receptor, since the exposure of cardiac tissue to ouabain in a Ca,+-free medium containing Na+ resulted in a development of the positive inotropic effect. Thus, ouabain does not interact with its inotropic receptor in the absence of Na+. The properties of the ouabain-receptor interaction resemble those of ouabain binding to Na+, K+-ATPase. In addition, ouabain increases the mobility of superficially bound Ca2+.


Multiregional cerebral blood flow changes induced by a cholinomimetic drug. The effect of parasympathomimetic drug, carbachol, on regional cerebral blood flow was tested in the rabbit with the 14C-ethanol tissue sampling technique. Intracarotid injection of carbachol significantly increased flow with respect to untreated control animals in 6 out of 10 structures sampled. However, compared with animals to which atropine was also administered, the flow increases were greater with carbachol alone. These effects are discussed in terms of muscarinic and nicotinic activation.


p-Chloroamphetamine antagonism of cobaltous chloride-induced hypothermia in mice. Serotonin levels were measured in the brains of mice at 2 h and 10 days following the administration of a single dose of 10 mg/kg, i.p. of p-chloroamphetamine HCl (PCA). A 44% reduction in the concentration of brain serotonin was noted 2 h after PCA injection. No significant change in brain serotonin was observed 10 days following PCA administration. The time course of the body temperature response to cobaltous chloride (25 mg/kg, i.p.) was recorded 60 min following pretreatment with PCA (10 mg/kg, i.p.). PCA reduced the hypothermic response by approximately 60%, presumably through the depletion of central 5-hydroxytryptamine.


Failure of opiates to reverse dopamine inhibition of prolactin secretion in vitro. To identify the site of action of opiate-induced prolactin elevation, in vitro rat hemipituitary incubations were performed in the presence of morphine, met-enkephalin, ala2-met5-enkephalinamide and dopamine (DA) and combinations of opiate with the catecholamine. No opiate stimulated prolactin release directly nor did any opiate block the inhibitory effect of DA. We conclude that this opiate endocrine action is not mediated at a pituitary level but may involve interference with hypothalamic DA release.


The failure of hydrogen peroxide to improve function in ischemically depressed myocardium. The hypothesis that hydrogen peroxide might lend inotropic support to ischemically depressed myocardium was critically tested in a canine model. A coronary branch was perfused at reduced flow to depress mechanical function. Since hydrogen peroxide infusion into the perfusate failed to alter the segment's contractile force, it was concluded that the previously described beneficial effects of hydrogen peroxide infusions are probably not related to a restoration of contractility in ischemic myocardium.


Interaction of a potential antimigraine drug (Org GC 94) with the vasomotor action of serotonin. The interaction of a potential anti-migraine drug (Org GC 94) with the vasomotor action of 5-HT in vitro in feline, canine and human intra- and extracranial arteries, as well as in vivo in the canine nasal vascular bed as been investigated. In the two in vitro preparations, i.e. using superfusion or bath techniques, the intracranial vessels were more sensitive to 5-HT vasoconstriction than the extracranial ones. As both the maximum contraction and the slope of the dose-response curves were reduced by increasing concentrations of Org GC 94, the antagonism of 5-HT did not involve competitive blockade of 5-HT receptors. The dilator response was tested in arteries brought to a higher tone with prostaglandin F2 alpha. 5-HT produced a dose-dependent dilatation which, like the vasoconstriction, was antagonized in a non-competitive manner by Org GC 94. Intra-arterial injections of 5-HT provoked nasal vasoconstriction and this response was clearly potentiated by Org GC 94 in low doses while higher doses inhibited the vascular response to 5-HT. The specific effect of Org GC 94 in vivo may be the potentiation of 5-HT-induced vasoconstriction. The hypothesis is discussed that the so-called anti-5-HT agents act in migraine patients as partial agonists of 5-HT, mimicking rather than antagonizing 5-HT.


Accumulation of drugs by resting or beating cardiac tissue. The rate and degree of accumulation of 12 neutral, anionic and cationic drugs were studied in resting and 2 Hz-stimulated isolated left auricles of the guinea pig. The uptake process was accelerated in muscles driven electrically. The time needed to reach equilibrium was related to the extent of accumulation which in turn corresponded with the lipophilicity of the drug. The frequency-induced changes in the kinetics of the accumulation disappeared under reduced mechanical activity of the muscle. It is suggested that the increased mechanical activity enhances drug disposition within the extracellular space, so that more drug is available for transmembrane penetration.


Elevation of histamine levels in rat and mouse tissues by the deacetylation of administered N-acetylhistamine. N-Acetylhistamine administered to mice and rats (200-800 mg/kg, i.p.) elevated tissue histamine levels to a considerable extent. N-Acetylhistamine was more potent to elevate histamine than was histidine, a precursor amino acid of histamine. From the present results and the distribution of N-acetylhistamine deacetylase in tissues, it could be concluded that the elevation of histamine was due to the deacetylation of N-acetylhistamine. In addition, a slight but significant amount of N-acetylhistamine was found in tissues of normal rats. The physiological role of the deacetylation of N-acetylhistamine is discussed on the basis of these results.


Effects of enkephalin analogue and naloxone on cat spinal cord dorsal root potentials. A systemically active enkephalin analogue, FK33824, given intravenously depressed dorsal root potentials in cat spinal cord. The negative DR V and positive DR VI, measured by computer, were both decreased; this effect was reversed by small doses of intravenous naloxone. Naloxone, given alone, with no previous analogue produced no changes in dorsal root potentials suggesting the absence of a basal enkephalin tone. A second injection of FK33824 was much less effective that the first dose. The results were discussed in relation to presynaptic mechanisms for analgesia: we proposed that FK33824 causes presynaptic inhibition by modulation rather than by depolarization of primary afferent fibers.


Analysis of the activation of dopamine metabolism by a serotonin uptake inhibitor. The potentiation of the effect of haloperidol on rat striatal homovanillic acid (HVA) and 3,4-dihydroxyphenylacetic acid (DOPAC) by a serotonin (5-HT) uptake inhibitor, CGP 6085 A, has been further investigated. The evidence that this effect of CGP 6085 A is related to its 5-HT uptake inhibitory properties is discussed. The potentiation was antagonized by scopolamine and baclofen, and disappeared more rapidly than the uptake inhibitory effects of CGP 6085 A, presumably because the effects of uptake inhibition were counteracted by a reduction in 5-HT synthesis. CGP 6085 A also potentiated the effects of fluphenazine and pimozide, but not those of a number of other neuroleptics. It also failed to restore the effect of haloperidol in animals treated chronically with the neuroleptic. These results appear to indicate that CGP 6085 A can only stimulate DA neurons if their activity is already above the resting level. The antagonism of this effect by scopolamine suggests that the site of action of the 5-HT uptake inhibitor is before a cholinergic neuron.


Decrease in muscarinic cholinergic response of the rat heart following treatment with 6-hydroxydopa. Pretreatment with 6-hydroxydopa (6-OHDOPA) at birth produced a decrease in the number of [3H]quinuclidinyl benzilate ([3H]QNB) binding sites in adult rat heart homogenates. The treatment caused hyposensitivity to acetylcholine (ACh) but did not alter the maximal negative inotropic action of ACh in isolated atria of the rat. These results suggest that 6-OHDOPA affects the negative inotropic response to ACh by modifying the receptor number or through an effect on a step between receptor activation and biological response.


The cardioactive properties of SC4453, a digoxin analogue with a C17 beta-pyridazine ring. SC4453 is a digoxin analogue with a pyridazine instead of a lactone ring in C17 beta. SC4453 was compared with digoxin with respect to effect on contractility and on activity of the sodium pump in guinea-pig isolated left atria stimulated at 3.3 Hz. The two glycosides stimulated the sodium pump at low concentrations and inhibited at high concentrations. At the time to peak inotropic effect, for a similar inhibition of the Na pump, the increase in systolic tension was higher with SC4453 than with digoxin, whereas the increase in diastolic tension was similar for both. These observations confirm that the inhibition of the Na pump is not the only mechanism responsible for the positive inotropic effect of cardiac glycosides.


The relationship between morphine, aspartic acid and L-asparaginase in rats. Morphine and aspartic acid were administered separately and in combination to 80 rats divided into 8 groups. Ten and 20 min following the injections, brain, liver and kidney L-asparaginase activity was determined. Morphine decreased brain and liver L-asparaginase activity and increased that of kidney. Aspartic acid completely antagonized the effect of morphine. Additionally 500 IU/kg L-asparaginase and 5 or 10 mg/kg morphine were i.v. injected into 56 rats divided into 5 groups. L-Asparaginase, which, in turn, increased motor activity, antagonized the morphine-induced hypoactivity and analgesia. These results support our previous findings.


Post-tetanic potentiation in the innervated smooth muscle preparation of the guinea-pig ileum. The phenomenon of posttetanic potentiation of electrically evoked twitch contractions in the myenteric plexus-longitudinal muscle of the guinea-pig ileum was observed when (1) post-tetanic inhibition had been prevented by naloxone, an opiate receptor antagonist, and (2) the twitches had been diminished by indomethacin, an inhibitor of prostaglandin synthesis. The origin of posttetanic potentiation at this muscarinic synapse appeared to be presynaptic.


[Sex difference and fine structure on epithelium cells of Bowman's capsule in mice (author's transl)]. Bowman's capsules with cuboidal epithelium were found in 87.2%, 95.5%, 75.4%, 96.7%, 98.8% and 91.2% in the kidney of adult male C57BL/6J-ob, JCL-ICR, dd, E1, ASK and KK mice, respectively, while in 57.9%, 6.7%, 7.0%, 2.8% and 9.1% in adult female C57BL/6J-ob, JCL-ICR, dd, E1 and ASK mice, respectively. The renal corpuscles with cuboidal epithelium in Bowman's capsules of male JCL-ICR mice increased gradually in number from 5 to 10 weeks age. The surface of cuboidal epithelium cells was shown to have long cilia and dense microvilli, which were similar to those seen in proximal uriniferous tubules.


[Effect of proximate environment on drug susceptibility of mice (author's transl)]. The present study was performed in order to elucidate the effect of proximate environment on drug susceptibility of mice. Three experiments were carried out independently. In the first experiment, mongrel and ddS mice produced under an unsatisfactory control of proximate environment were purchased, and acute toxicity tests of thiamine hydrochloride (B1HCl) and isonicotinic acid hydrazide (INAH) were practiced at two different conditioned rooms. In the second experiment, ddY mice produced under the conventional environment controlled to a certain extent were purchased, and the toxic effect of B1HCl was examined under the similar environment. In the third experiment, the sensitivity to B1HCl of RFVL mice produced under the strict barrier system was tested at the severe air-conditioned room. LD50 and their fLD50 values were calculated by Litchfield-Wilcoxon's method, and the variance analyses were carried out. The severer the environmental control after the purchase of mice turned to the higher the drug sensitivity. This respect was more remarkable in INAH of which the toxic response is appeared slowly compared with B1HCl. Furthermore, seasonal variation was found in LD50 values. However, seasonal effect differed from rearing and experimental conditions. In the third experiment which these proximate environments were controlled severely, seasonal variation was very small. From the results of these experiments, it was defined that the use of animals produced under the satisfactory rearing condition and severe environmental control are necessary for animal experiments.


Genetic control of blood pressure in spontaneously hypertensive rats (SHR). Genetic control of blood pressure in the SHR strain was studied by three separate experiments which consist of cross analysis between the SHR and Donryu, two-way selecton for high and low blood pressure levels, and successive backcrosses to the parental strains. The results obtained were as follows. 1. The data from genetic crosses between the SHR and Donryu showed the phenotype segregation ratio of 1:1 at the backcross and 1:2:1 at the F2 generation. 2. Two-way selection for high and low blood pressure levels was performed from the F2 generation onward. The separation between the two lines occurred immediately after the first selection. Thereafter, the difference increased gradually with generation. The blood pressure level at the seventh generation of selection became approximately equal to those of the parental strains. 3. Two types of the successive backcross were performed from the F1 hybrids by mating the males showing the highest blood pressure level to Donryu females and the females showing the lowest blood pressure level to SHR males on the other. Bimodality was observed in the distribution of blood pressure levels at each generation. Their phenotypic segregation ratios were accordant with 1:1 on the whole. At the intercross generation during successive backcrosses, a trimodal distribution was observed. 4. These results confirmed that the hypertensive trait of the SHR is regulated by a single major gene and other several genes with minor effect. A gene symbol ht was proposed for this major gene. Concurrently, a congenic strain having the ht gene on the genetic background of the Donryu was developed by the successive backcross system.


[Comparative studies on release of enzymes from platelets during blood clotting in Rattus rattus flavitecus and Rattus norvegicus (author's transl)]. Increase of lactate dehydrogenase (EC 1.1.1.28) and creatine kinase (EC 2.7.3.2) in serum and plasma level during blood clotting was compared in R. rattus flavitectus (black rat) and R. norvegicus (albino rat). Both the enzyme level increased by the release from platelets in process of clotting in rat as previously described (Emori T., Takahashi M. and Nagase S. (1978) Exp. Animals, 27, 167), but the change in black rat was very slight. Owing to search for the reason to prove the difference in this phenomenon, physiological and chemical properties of blood from both the animals were compared, and it was demonstrated that the content of plasma fibrinogen in black rat is higher than that in albino rat, while the platelet number contained in each ml of blood and the enzyme activities in each platelet are not different between them. The analysis of the membrane proteins from platelets of both the animals by SDS-polyacrylamide gel electrophoresis showed that the fractional value of glycoproteins was found to be different between them.


The age-related tumor incidence in Wistar-Imamichi rat. The age-related incidences and types of spontaneous tumors were studied in 320 male and 354 female Wistar-Imamichi rats reared up to 130 weeks of age. The tumor incidence increased abruptly between 53 and 78 weeks of age, and 170 males and 269 females were shown to bear tumors during the observation period. Multiple tumors occurred in 62 males and 115 females. Most common were hypophyseal (male 69: female 224), mammary gland (29:99), testicular (74:0) and adrenal gland (26:21) tumors. The incidences of hypophyseal and testicular tumors increased with age. Tumors occurred in later stages of the observation period in the ovary, uterus and vagina (0:27), thyroid and parathyroid (13:12), skin (8:8), spleen and lymph nodes (8:7), pancreas (9:5), liver (3:7), lung (3:3), thymus (1:4), omentum and mesenterium (2:3), stomach and intestine (2:2), kidney and urinary bladder (1:2), right femur (0:1), and brain (0:1).


Characterization of a Friend virus-replicating target cell. The FV-replicating target cell characterized in this study is an erythroid precursor cell, as shown by its response to bleeding and hypertransfusion. The target cell is not identical with the BFU-E compartment as demonstrated by different model velocities and different cell cycle characteristics as compared to BFU-E. Different sensitivity of FV-target cells and CFU-E to AMD, differences in the growth kinetics of both cell populations in bone marrow and spleen of mice after bleeding, and large quantitative differences of both cell populations in hemopoietic organs, suggest only partial identity or non-identity  of these two cell types. Finally it is shown that spleen colonies originating from FV-infected target cells are a primary erythroleukemic lesion, since they contain more Ep-independent CFU-E than do intercolony areas of the spleen.


Surface antigens of murine hemopoietic stem cells. I. Cross reactivity of antisera against differentiated hemopoietic cells with bone marrow stem cells. A model of multiply marked hemopoietic stem cells proposed by Till (1) has been tested with respect to antisera raised against differentiated murine hemopoietic cells. When absorbed with erythrocytes, antisera against CBA mouse lymph node lymphocytes, thymocytes, peritoneal macrophages and platelets cross-reacted strongly with pluripotent stem cells (CFUs) in bone marrow as determined by inhibition of spleen colony formation in lethally irradiated mice. Absorption of ATS, antimacrophage serum and antiplatelet serum with hemopoietic cells other than those used to prepare the antisera (e.g., ATS with neutrophils and platelets, antimacrophage serum with neutrophils, thymocytes and platelets and antiplatelet serum with neutrophils and thymocytes) did not reduce the activity of these antisera for CFUs whereas absorption with the inoculating cell type greatly reduced anti-stem cell activity. Absorption of these antisera with non-hemopoietic tissues such as brain, kidney, liver and testis in general had little effect on antistem cell activity, although a significant loss of activity was observed following absorption of antiplatelet serum with kidney. The antistem cell activity in ATS, antimacrophage serum and antiplatelet serum does not appear to be caused by antibodies against histocompatibility antigens sine bone marrow stem cells from histoincompatible C57BL and Balb/c mice were also sensitive to antisera against CBA mouse hemopoietic cells. In contrast to these findings, antisera against erythrocytes showed little cross-reactivity with CFUs, indicating that few antigens are held in common between erythrocytes and CFUs. We propose that nucleated hemopoietic cells and platelets retain cell line specific antigens in common with pluripotent stem cells from which they were derived, and that the continued expression of these antigens during differentiation may be involved in the differentiation process.


Turnover of circulating hematopoietic stem cells. Short-term parabiosis of male and female CBA/CaJ mice was used to investigate the turnover of circulating hematopoietic stem cells. The exchange and subsequent disappearance of donor stem cells were monitored by spleen colony assay and chromosome analysis of individual colonies. The results revealed an exponential disappearance of pluripotent stem cells from blood with a characteristic half time of 1.7 h. Blood-borne stem cells were shown to be equilibrated with a subpopulation of marrow stem cells exhibiting a disappearance half time of 9.5 h. Splenectomy did not change the apparent rate of stem cell removal from the blood.


Thyroid hormones and protein synthesis in fetal mouse liver erythroid cells. Hemoglobin synthesis represents an important step in the maturation of the developing erythron. In this study, we determined the effect of various thyroid hormones and analogs upon globin synthesis by fetal mouse liver erythroid cells. Globin synthesis was stimulated in 13--15 day fetal liver erythroid cells, whereas cells, whereas cells from 17 day fetuses were unresponsive. There was no correlation between stimulation of globin synthesis and reported calorigenic activity of the various hormones assayed.


Morphological and biochemical alterations of the lung after application of chlorphentermine. The influence of short-term and chronic administration of chlorphentermine (Chlph) on the structure and phospholipid (PL)-content of the lungs was studied in guinea pigs. The drug was given daily in a dose of 40 mg per kg body weight i.p. The PL-content of the whole lung as well as the phosphatidyl choline (PC)-portion were estimated in animals treated with Chlph for 4, 5 and 6 weeks. In a further experiment, the influence of 2, 4 and 28 days application on the PL-content of the lung lavage fluid separated into the acellular and cellular (macrophage) fraction was examined. Morphologic studies of lung, liver, heart, kidney, spleen, brain, and pancreas supplemented the PL-content analysis. The amount of PL in the whole lung increases continuously with the duration of the treatment but only moderately. The PC-fraction of the total PL remains constant throughout the experiment. A substantially greater increase was detected in the lung lavage fluid whereby the cellular fraction is most affected. Very important is the result that the PL-content of the lung lavage fluid reaches its highest level already after two Chlph-injections. After short-term administration the morphological examination of the lung mainly shows inflammatory infiltrations, whereas after long-term application a foam cell reaction is mainly localized in the interstitium. The studies show that an increase of the amount of lung sufactant in guinea pigs is possible following Chlph administration.


Antiinflammatory substances appearing during an acute inflammation and the alteration of their properties by isoproterenol (ISO). An in vivo investigation in rats. It could be demonstrated that antiinflammatory substances appear during the course of an acute inflammation. The inhibition of leucocyte immigration into the site of inflammation (caused by Sephadex) was taken as criterion for the antiinflammatory property. The highest quantity or effect of the antiinflammatory substances was found on the 3rd day after inflammation has started. Not only the absolute number of cells but also the composition of the cellular exudate (compared with a fresh inflammation) was changed: The percentage rate of the neutrophils was reduced, whereas the portion of the eosinophils and the lymphocytes was increased. A permanent ISO application on the animals altered the quantity of the composition of these substances, in consequence of which the immigration of eosinophils and lymphocytes was promoted. The reaction of the neutrophils was unaffected.


Myelin lipids in Wallerian degeneration of the rabbit optic nerve. Wallerian degeneration of the rabbit optic nerve was produced by enucleation and the myelin output as well as its lipid composition were studied. In spite of the marked drop in the myelin mass, occurring very soon after enucleation the lipid composition of the corresponding myelin fractions did not change appreciably, phosphatidylcholine being the most resistant lipid species. However, from the very beginning of the degenerating process there appeared in the myelin lipid spectrum marked amounts of cholesteryl esters and phosphatidic acids and in the later period elevated amounts of lysophosphatidylcholine. Enhanced esterification of cholesterol could function as an early primary factor injuring the myelin membrane, the lysocompounds may be involved at a later stage in the pathomechanism of myelin decomposition in the central nervous system.


Effects of serum concentrations on the clonal growth of Syrian golden hamster fetal lung cells. In order to gain an insight into the mechanisms of clonal growth of early-passage Syrian golden hamster fetal lung (SGHFL) cells, a study was undertaken, in which the effects of various serum concentrations on the cloning efficiency and on the colony size of these cells were examined. The results suggest that both cloning efficiency and colony size are influenced by at least two factors present in the serum.


Egocentrism and inefficiency in the communication of families containing schizophrenic members. Conceptual and methodological shortcomings of research on family and interactional psychopathology are owing mainly to the use of vague and ill-defined concepts of communication. Based on a theory of language use and communication within general social and cognitive psychology (e.g., Heider, Mead, Piaget, Rometvet) Blakar has outlined a methodology by which interaction is analyzed in terms of how and to what extent the participants (families) manage or fail to cope with the various prerequisites for successful communication under varying situational conditions. A study illustrating this program is presented: The interaction of twelve families, six with (Group S) and six without (Group N) a schizophrenic member, is analyzed in Blakar's communication conflict situation with respect to the members' ability to decenter and take the perspective of each other. Group S proved significantly more egocentric, their egocentric attitudes resulting, as would be expected, in very inefficient communication. Moreover, Group S were not able to adapt their pattern of communication to the changing situational requirements. Finally, the subtle interplay between the capacities and behavior of the individual members and the family system is illustrated: the egocentrism of the members resulted in "closed systems," and the closed systems hindered adequate feedback, forcing the members to decenter.


The relative value of empirical evidence. Arguments between adherents of different therapeutic models are often based on the assumption that empirical evidence is ultimately the judge of the merits of the respective approaches. The gathering of empirical evidence, however, is programmed by epistemological premises that in turn are not tested by "objective" demonstration but rather by their congruence with prevailing sociocultural values. The epistemological premises behind indivdual- and systems-oriented models are incompatible, and respectively congruent with currently coexistent and opposite sociocultural values. Adherence to a particular model is thus based on epistemological-axiological positions rather than neutral objective evaluation.


A study of the relationship between family interaction and individual symptomology over time. The study was designed to answer two questions: (a) Can a relationship over time between family interaction and individual symptomatology be demonstrated? (B) Can it be shown that changes in interaction have more influence on changes in the symptom than vice versa. Five interaction dimensions were taped in weekly, three-quarter hour sessions over 20 weeks in five families, each consisting of a mother-father-child triad who met certain criteria and had an encopretic child. The interaction dimension scores were abstracted weekly from these sessions by content analyses relying on various scales. The derived ratio scores were then related to the number of days the child soiled himself in the week preceding and following the interaction measurement. The relationship was assessed by Pearson correlation coefficients and step-wise multiple regression analyses adjusted to account for the possible inflationary effects of taking measures from the same subjects more than once. The results of the analyses answered both questions posed by the study in the affirmative, thus supporting the rationale underlying family therapy.


The family therapist behavior scale (FTBS): development and evaluation of a coding system. This study evaluated the validity and reliability of a new coding system--The Family Therapist Behavior Scale (FTBS)--that was designed to identify and study clinically relevant verbal behaviors of short-term, problem-oriented family therapists. Validity was assessed by testing the scale's ability to discriminate significant, predicted differences between the in-therapy behaviors of eight beginning family therapists conducting observed interviews and eight advanced family therapists conducting supervisory interviews. All of the sessions, which were initial interviews, were videotaped. Two coders rated three five-minute samples from each of the 16 tapes with the FTBS. The validity results supported over 50 per cent of the 16 research hypotheses. The reliability analysis, based on the actual study data, indicated that the interrater reliability of the 19 category FTBS differed from chance at less than the .001 level of significance. The implications of these findings are examined and future research directions are identified.


Direct open supervision: a team approach. Created in the absence of one-way mirror facilities, Direct Open Supervision combines elements of live supervision with team participation in training family therapists. The theory and application of this approach to direct supervision are described.


The effects of sibling death on the surviving child: a family perspective. The death of a child invariably affects the family, who in effect become survivors. Adaptations are made in order to secure a new family equilibrium attendant upon such a loss. Surviving siblings not infrequently become the focus of maneuvers unconsciously designed to alleviate guilt and control fate through silence and efforts to maintain silence, through substitution for the lost child, and through endowing the survivor-child with qualities of the deceased. Three types of clinically identifiable types of survivor-children are described. Families that emphasize silence and focus on guilt, families in which the child becomes incomparably precious, and families in which substitution and replacement provide the major theme lead respectively to the "haunted," "bound," and "resurrected" child. These children share many features, as do their families, but there appears to be a connection between the family defensive maneuver and the specific consequences for a child of the bereaved family.


Using systems theory to organize confusion. This paper represents an initial step in applying to complex clinical situations the symbolic logic developed by G. Spencer Brown and elaborated by Francisco Varela. This way of modeling turns out to yield an interesting mixture of rigidity, ambiguity, and paradox, perhaps inevitable at our present level of understanding of systems containing feedback. Applying the Brown-Varela concepts seems a useful transitional step toweard the future use of more sophisticated quantitative models such as those of Powers and Forrester.


[Antimicrobial activity of derivatives of 1,2,4-benzothiadiazine-1,1-dioxide. VIII]. The antimicrobial activity of a series of fluoro derivatives of benzothiadiazine and sulfonamides was studied. The compounds tested can be grouped as: a) 3-alkylmercapto derivatives of 6-trifluoromethyl-1,2,4-benzothiadiazine-1,1-dioxide (III leads to VI); the 3-mercapto precursor (VII) and the related 3-picolinic salt (VIII); b) 3-trifluoromethyl derivatives of 1,2,4-benzothiadiazine-1,1-dioxide and of its benzene substituted derivatives (IX leads to XVI); c) trifluoroacetylaminobenzenesulfonamides (XVII leads to XXV). Two of the 3-alkylmercapto compounds [(V) and (VI)] showed marked inhibitory activity against some strains of Staphylococcus, Streptococcus and Diplococcus. None of the compounds tested proved active against Gram-negative schizomycetes (genera Salmonella, Shigella, Escherichia, Proteus, Pseudomonas, Enterobacter, Klebsiella, Serratia, Yersinia, Providencia) or against yeasts (Candida).


[Heteroarylalkanoic acids with potential anti-inflammatory activity]. A series of (3-oxodihydro-1,2,4-benzothiadiazin-1,1-dioxide-3-yl)acetic acids [compounds of type (A)] and (1,2,4-benzothiadiazin-1,1-dioxide-3-yl)oxyacetic acids [compounds of type (B)] were synthesised and tested for antiinflammatory activity. Preliminary tests showed certain compounds to have a significant level of antiinflammatory activity in rat paw edema induced by carrageenan. It was found that the antiinflammatory activity of this series of compounds depends on the nature, number and position of the substituents on the benzene ring.


In vitro action of uridine diphosphate glucose (UDPG) on phrenic diaphragm preparations. The action of uridine-5'-diphosphoglucose (UDPG) on the contractile response of the phrenic diaphragm preparation from guinea pig was investigated. UDPG activity was assayed on the preparation at rest or during the exercise; in this case indirect electrical stimulation of phrenic diaphragm preparation or direct stimulation of denervated or curarized muscle were employed. Krebs' solutions adequately modified with regard to glucose concentration were used. The effect of UDPG on the neuromuscular junction was also investigated by recording miniature end plate potentials. An effect of the drug on neuromuscular transmission and on glucose metabolism could be demonstrated.