Catechol, a structural requirement for (Na+ + K+)-ATPase stimulation in rat skeletal muscle membrane. 1. Catecholamines can nearly double (Na+ + K+)-ATPase acts effect is not mediated by cyclic AMP and is not beta-adrenergic. 3. Orthodihydroxybenzene compounds and their orthoquinone derivatives enhance (Na+ + K+)-ATPase activity. 4. Enhancement of (Na+ + K+)-ATPase activity by catechols is not due to increased availability of ATP. 5. It is suggested that catechols and their orthoquinones somehow alter or protect the configuration of the enzyme so that it becomes more active or so protect the configuration of the enzyme so that it becomes more active or so that its activity is maintained under conditions in which its activity is otherwise diminished.


beta-Adrenergic effect on Na+-K+ transport in rat skeletal muscle. 1. Intact rat extensor digitorum longus muscles soaked in L-isoproterenol plus 10(-5) M ouabain gained less sarcoplasmic Na+ than did muscles soaked in ouabain alone. Half maximal effect was produced by 10(-8) M L-isoproterenol. 2. D-Isoproterenol and oxidized L-isoproterenol were only 3 and 1%, respectively, as potent as L-isoproterenol. Other catechols tested had no effect. 3. The effect of L-isoproterenol on sarcoplasmic Na+ content appears to be a beta-adrenergic function in that it was blocked by propranolol, but not by phentolamine, and could be mimicked by dibutyryl cyclic AMP or by caffeine. 4. Reduced gain in sarcoplasmic Na+ was accompanied by reduced loss of sarcoplasmic K+. 5. L-Isoproterenol increased loss of sarcoplasmic Na+ in the absence of ouabain, in muscles recovering from cold treatment. 6. Results suggest that the beta-adrenergic system stimulates a coupled Na-K+ pump. 7. A model is proposed in which stimulation of the Na+-K+ pump in response to beta-adrenergic agents involves a number of intermediate steps, identified tentatively.


A spin label study of lipid oxidation catalyzed by heme proteins. Rapid loss of the electron spin resonance signal from a variety of spin labels is observed when ferricytochrome c or metmyoglobin are combined with lipids. Evidence is presented that this loss of signal can be used as a sensitive method to study lipid oxidation catalyzed by heme proteins. Under aerobic conditions and with lipids which bind the heme protein, the kinetics of the oxidation process as observed by the spin label method are identical to the kinetics previously observed by measurements of oxygen uptake. Use of pre-oxidized lipids under anaerobic conditions indicates that cytochrome c reacts with a product of lipid oxidation. Kinetic studies of the anaerobic reaction indicate that cytochrome c reacts rapidly with lipid oxidation products in membrane areas far larger than the area occupied by cytochrome c, implying rapid transport of reactive species within the membrane interior in directions parallel to the membrane surface. Under anaerobic conditions, reaction of cytochrome c with lipid oxidation products appears to produce a relatively long lived (hours) species located in the hydrophobic portion of the membrane, which is capable of subsequent reaction with lipid-soluble spin labels.


Location of two sulfhydryl groups in the rhodopsin molecule by use of the spin label technique. Sulfhydryl groups of membrane-bound rhodopsin are studied with the spin label technique by using five maleimide derivative probes of different lengths. Two sulfhydryl groups are titrated per molecule of rhodopsin, These groups are located in protected but probably different environments, less than 12 A away from the aqueous phase. A distance of about 37 A is measured between the two groups. These results are consistent with a model in which the two groups would be located close by the external surface of the protein but embedded within the membrane layer, the strong immobilization of the label molecules resulting from phosphlipid-protein interactions.


Electron spin resonance studies of the membranes of the cellular slime mold Dictyostelium discoideum. Doxylstearic acid spin labels are used to study the fluidity of the membranes of the cellular slime mold, Dictyostelium discoideum. The tau omicron value of the wild-type cell membrane is close to that of egg lecithin indicating a rather fluid membrane. No detectable change in the fluidity of the bulk lipids at the 16-carbon depth occurs during differentiation of the myxamoebae into stalk and spore cells despite reported changes in the individual lipid components. The results of studies on temperature-sensitive and aggregationless mutants are also presented.


Studies on the iodinated surface membrane proteins and concanavalin A agglutination of transformed Syrian hamster cells. Chemically transformed Syrian hamster cells exhibit marked agglutination in the presence of the plant lectin, concanavalin A. In this report, we describe conditions which can alter this concanavalin A agglutinability, and compare the surface proteins from transformed cells which express different degrees of agglutinability. Lactoperoxidase-catalyzed iodination of tertiary Syrian hamster cells reveals the major iodinatable protein to be approximately 220 000 daltons. The transformed Syrian hamster cells do not contain this protein in an iodinatable form. Analyses of the transformed cells grown under conditions which decrease the concanavalin A agglutinability do not demonstrate any iodination of the 220 000 mol. wt. protein. These results depict the effects of growth and dibutyryl cyclic AMP on the iodinatable cell surface proteins of transformed cells and indicate that the absence of the I-220 000 mol. wt. protein is probably not a major determinant of concanavalin A agglutination.


Divergent effects of cyanate on amino acid and phosphate uptake by liver and hepatoma. The uptake of alpha-aminoiso[3H]butyric acid and 32Pi was observed to be inhibited by sodium cyanate in transplanted hepatomas but was increased in the livers of the tumor bearing rats. Incorporation of 32Pi into macromolecules in hepatomas was also inhibited by cyanate. Treatment with this drug did not influence circulating concentrations of isotope-labeled materials. There were relatively small effects on uptake of 36Cl- in cyanate-treated rats and the action was not tissue specific. The data were compatible with an inhibitory effect of cyanate on active transport in hepatomas which was not seen under the same conditions in host liver.


Thermal elution chromatography of nucleic acids on hydroxyapatite. Hydroxyapatite thermal elution chromatography was studied from an empirical standpoint. The dependence of elution temperature on elution buffer concentration was determined for various types of buffer, hydroxyapatite and nucleic acid. The results are analyzed in terms of the proper design and interpretation of thermal elution experiments. The potential for serious artifacts is demonstrated and the means by which they may be avoided is described. Various commercially available hydroxyapatites were tested in conjunction with various aqueous and partially non-aqueous buffer systems. Among the materials tested, potassium phosphate and Bio-Rad HTP were found to constitute the best buffer-hydroxyapatite system for most types of thermal elution study.


Properties of the activator-dependent cyclic nucleotide phosphodiesterase from bovine heart. In the absence of activator, the activator-dependent cyclic nucleotide phosphodiesterase (EC 3.1.4.-) from bovine heart hydrolyzed 3.2-fold more cyclic GMP than cyclic AMP when assayed with 10(-6) M substrate in the presence of 5 mM Mg2+ and 10 muM Ca2+. The addition of saturating amounts of phosphodiesterase activator increased the hydrolysis of cyclic GMP 8-fold and cyclic AMP 6-fold. The pH maxima of the enzyme was rather broad from 6.0 to 7.0 for the hydrolysis of both cyclic GMP and cyclic AMP in the absnece or presence of phosphodiesterase activator. Substrate specificity of the enzyme in the absence or presence of phosphodiesterase activator varied depending on the divalent metal used to support activity in the absence of activator. The enzyme preferentially hydrolyzed cyclic GMP in the presence of Mg2+ while little substrate specificity was observed in the presence of Mn2+, Zn2+, Co2+ or Ni2+. The magnitude of the increase in enzyme activity due to activator and Ca2+ varied depending on the divalent metal used to support enzyme activity without activator. The addition of activator increased the V of cyclic AMP hydrolysis 1.7-fold while causing no significant change in the Km for cyclic AMP. Two apparent Km values for cyclic GMP (1 and 15 muM) were noted in the absence of activator and upon the addition of activator a single apparent Km (3 muM) was observed with a V approx. 2-fold above that in the absence of activator. Cyclic AMP competitively inhibitied the hydrolysis of cyclic GMP with a ki of 50 muM while cyclic GMP non-competitively inhibited the hydrolysis of cyclic AMP with a Ki of 1.8 muM. The results suggest there may be two or more binding sites for cyclic GMP on the enzyme and possibly only one binding site for cyclic AMP.


Structural and kinetic studies on the activators of succinate dehydrogenase. 1. Diverse classes of compounds such as dicarboxylates, pyrophosphates, quinols and nitrophenols are known to activate mitochondrial succinate dehydrogenase (EC 1.3.99.1). Examples in each class -- malonate, pyrophosphate, ubiquinol and 2,4-dinitrophenol -- are selected for comparative studies on the kinetic constants and structural relationship. 2. The activated forms of the enzyme obtained on preincubating mitochondria with the effectors exhibited Michaelian kinetics and gave double-reciprocal plots which are nearly parallel to that of the basal form. On activation, Km for the substrate also increased along with V. The effectors activated the enzyme at low concentrations and inhibited, in a competitive fashion, at high concentrations. The binding constant for activation was lower than that for inhibition for each effector. 3. These compounds possess ionizable twin oxygens separated by a distance of 5.5 +/- 0.8 A and having fractional charges in the range of -0.26 to -0.74 e. The common twin-oxygen feature of the substrate and the effectors suggested the presence of corresponding counter charges in the binding domain. The competitive nature of effectors with the substrate for inhibition further indicated the close structural resemblance of the activation and catalytic sites.


Spectrophotometric studies on NAD(P)H oxidase of leukocytes. 1. The relationship between granule-NAD(P)H oxidase and myeloperoxidase. The NAD(P)H oxidase located in granules from resting leukocytes seems to be identical with myeloperoxidase on the basis of the following results. Spectral changes representing the difference between granules with and without NAD(P)H under various conditions represented the formation of compound III of myeloperoxidase, corresponding to the oxidation of NAD(P)H. The KCN difference spectrum of granules from both resting and phagocytizing leukocytes was in agreement with the KCN difference spectrum of myeloperoxidase. The affinity of KCN for myeloperoxidase was the same in both resting and phagocytizing leukocytes. The KCN-sensitive portion of NAD(P)H oxidase of granules from phagocytizing leukocytes seems to be identical with isolated myeloperoxidase and the myeloperoxidase of resting leukocytes. The KCN-insensitive oxidation of NAD(P)H by granules from phagocytizing leukocytes has not been found to be identical with myeloperoxidase.


The binding of [3H]dolichol by plasma high density lipoproteins. Radioactive dolichol was rapidly taken up by a plasma protein fraction following the intravenous injection of detergent suspended [3H]dolichol into rats. The half-life of labeled dolichol in the plasma was found to be approx. 13 h. Density gradient centrifugation or gel filtration of samples of human or rat plasma incubated with [3H]dolichol suspensions also showed that the labeled material became associated with the same plasma protein fractions as in the in vivo studies. Evidence was obtained to indicate that dolichol is specifically associated with the high density lipoprotein fraction of plasma. The possible role of high density lipoproteins as a vehicle for dolichol transport and the problems involved in the binding of this long moleculare are discussed.


Neuraminic acid-specific modification and tritium labelling of gangliosides. 1. A crude ganglioside mixture and pure GM1 and GD1a from bovine brain grey matter were prepared on a large scale. 2. The C7- and G8-analogues of NeuNAc were prepared from Collocalia mucoid and their structures established by gas-liquid chromatography and mass spectrometry. 3. Using model compounds in addition to various gangliosides, the conditions for the periodate oxidation and subsequent borohydride reduction of gangliosides were investigated with regard to the yield of C7- and C8-analogues of NeuNAc and the integrity of other monosaccharides in the oligosaccharide chain. These conditions were optimised to yield maximum C8-NeuNAc production and low C7-NeuNAc formation. Thus products were obtained which closely resemble the native gangliosides. 4. Using boro [3H] hydride, ganglioside derivatives with high specific radioactivity were prepared for the first time, containing either NeuNAc and labelled C8-NeuNAc or mainly labelled C7-NeuNAc depending on the prevailing conditions.


Study of the long chain bases of sphingomyelin of Entomophthora coronata. Sphingomyelin was isolated from a strain of Entomophthora coronata, a fungus pathogenic for humans. After hydrolysis, the long chain bases were converted to their dinitrophenyl (N2pH) derivatives and the aldehydes prepared by oxidizing these compounds with periodic acid. The aldehydes were studied by gas chromatography. Twelve different aldehydes were identified, the chain distributiion ranging from C14 to C17. The prominent chains were unsaturated. Straight and branched chains were found. The most abundant parent base which formed 52% of the total aldehyde was a 1,3-dihydroxy-2-aminohexadecene.


Interaction of apoliprotein C-III with phosphatidylcholine vesicles. Dependence of aproprotein-phospholipid complex formation on vesicle structure. We have studied the interaction of an apolipoprotein from human very low density lipoproteins (apoC-III) with egg yolk phosphatidylcholine in the form of single- and multi-bilayer vesicles. The reactivity of single-bilayer vesicles with apoC-III appears to be greater than that of the multi-bilayer vesicles according to several thermodynamic and spectrosconic criteria. In the complexes formed by the association of apoC-III with single-bilayer vesicles, the alpha-helical content of the peptide backbone and the apolarity of the environment around the tryptophan residues are greater than that observed in the complexes formed with the multibilayer vesicles. A higher yield and more homogeneous density distribution of lipid-apoprotein complexes results from the interaction of apoC-III with the single-bilayer vesicles relative to those obtained with the multi-bilayer vesicles. The enthalpy of association of apoC-III with phospholipid was greater for the single-shelled vesicles (25 kcal/mol apoC-III) than for the multi-shelled ones (18 kcal/mol apoC-III). The difference in reactivity of these two types of liposomes is not due to a difference in their fluidities since their fatty acid compositions are identical, but may be due to a difference in their areas of sterically accessible phospholipid, their permeabilities to the apoprotein, their radii of curvation, or a combination of these factors.


Control of long chain fatty acid oxidation in heart mitochondria as studied by spin labeling. Spin-labeled stearic acid is shown to exhibit the same beta-oxidation kinetics as normal stearic acid. ESR spectra recorded in conditions allowing beta-oxidation indicate that membrane-bound fatty acids can be directly beta-oxidized and that the rate of this reaction depends on the concentration of albumin in the medium. The regulating function of albumin and pool role of the lipidic phase of the mitochondrial membranes are discussed.


[Possible role of free radicals in the transformations and mechanism of physiological action of vitamin D]. Deficiency of vitamine D is accompanied with a decrease of ESR signal intensity from g = 2.003, as well as of the signals with g = 1.94 and gav. = 2.25 in the liver and kidneys of young rats. Injection of 2.5 g ergocalciferole (vitamine D2) to these animals results in a complete or partial normalization of the intensity of these signals during 6--16 hrs and an increase of the ESR signal intensity from g = 2.003 in small intestine mucosa.


[EPR and low temperature spectrophotometric study of the phototransformation products of bacteriorhodopsin]. It is shown that BR and intermediate products of its phototransformation P600, P550 and P415 (the maximum at -196 degrees C at 419 nm) are not paramagnetic. Illumination of samples containing P415 (P419) at -- 196 degrees C with light in the region of 360-480 nm results in the formation of paramagnetic centres with a sunglet spectrum deltaH=18 Oe and g=2.002 (R1). In parallel formation of a new photoproduct P421 in the absorption spectrum is observed. During subsequent heating at -140 degrees C formation of an asymmetric signal with deltaH=45 Oe and g=2.006 and g=2.03 was observed. In the absorption spectra a dark transition. P421-P565 was observed under the same conditions. P565 differs from initial BR P570 as to its photochemical properties. R1 is identified as retinal radical, R2 as a peroxide radical of the BR-complex lipids. Paramagnetic, spectral, and photochemical properties of some products of BR transformation are compared. A scheme of oxidative-phosphorylation processes with participation of Mn ions in BR phototransformation.


[Complex formation of spin-labeled trideoxyadenylate with polyribouridylic acid]. It is shown that the ESR spectra of triple-stranded complex formed by the polyribouridylic acid with the trideoxyadenylate spin-labelled on terminal phosphate is a superposition of the signals with strongly different width of HFS-components. This result is a structural support of the early proposed supposition about the irregular relative orientations of the oligomers in the triple-stranded complexes of oligoadenylates with poly-U.


[Mechanism of oxidative phosphorylation. I. Role of the respiratory chain]. Mitochondria of white rat brain increased the respiration on succinate in response to ADP addition; in the presence of after NADH the respiration remained unchanged or decreased ofter addition ADP. When organelles were suspended in water distilled in the concentration of 3 mg of protein/ml and kept at melting ice temperature for 24 hours the response of mitochondria to ADP was not changed. Stechiometric relation between the number of electron of the oxidized substrate and absorbed by oxygen depended on ADP during succinate oxidation and did not change on NADH. ADP phosphorylation is suggested to proceed on the stage of the substrate dehydrogenation, rather than on the cytochrome part of the respiratory chain.


[Spike tranfer in statistical neuron assemblies. II. Neuron--nonlinear spike source]. The mathematical model of the neuron function is known to rely on space summing of excitement. The spikes contribute to the inner state of the neuron the farther from cell soma the synapses are located. The difference between excitatory and inhibitory effect results in spike firing if only neural firing threshold is achieved. The values of spike flux have been estimated on the basis of the model of CA3 sector of the Hippocampus and were found to be 15 divided by 35 imp/s.


[Left-helical conformation (type poly-L-proline II) of the polypeptide chain of adrenocorticotropic hormone in water]. By circular dichroism method the nature of the secondary structure of owine and bovine adrenocroticotropic hormone in water has been studied. An analysis of circular dichrosim spectra and their dependence on temperature point to a left-helical conformation (of the poly-L-proline II type) of the polypeptide chain of adrenocorticopic hormone in water.


[Interaction of spin-labeled saturated and unsaturated fatty acid derivatives with biological membrane lipid and protein components]. Interaction of saturated and unsaturated derivatives of spin-labelled fatty acids (2,2,6,6-tetramethyl-4-palmitoylpiperidil-l-oxyl (I) and synthetized by us 2,2,6,6-tetramethyl-4-oleoylpiperidil-1-oxyl (II) with sarcoplasmic reticulum membranes, retina rod outer segments and erythorocyte ghosts was studied. ESR spectra of the probe I are practically unresolved singlets, whereas ESR spectra of the probe II appear as more of less resolved triplet signals. The difference in ESR spectra of the probes I and II is the result of distribution of the probe I in pure lipid regions of membranes, the probe II being located both in lipid bilayer and hydrophobic areas of proteins.


[Localization of unpaired electrons in molecules of lysozyme substrate-inhibitors. I. N-acetylglucosamine]. Electron structure of N-acetylglucoseamine molecule and its ion-radicals has been studied by quantum-chemical methods CNDO2 and JNDO, and by ESR as well. N-acetyl group is shown to be the only electron acceptor group of AGA molecule. Phototransformations of paramagnetic centres are studied at different pH values.


[IR-spectroscopic study of lecithin and cholesterol layers adsorbed on Aerosil]. The IR-study of adsorbed layers of lecithin and cholesterol on aerosil was carried out. It was found that lipid molecules interact with adsorbent by means of hydrogen bonds with free surface hydroxyls. The structure of lipid layers on the surface of aerosil is discussed.


Formation of lipid inclusions in normal human leukocytes. Normal peripheral blood leukocytes left standing at room temperature develop large inclusions which increase in number and size with time of incubation. The formation and nature of these inclusions were investigated. At 0 hour the structures were present in only 1% of the cells, whereas at 48 hr, they were in virtually all neutrophils and monocytes. Ultrastructurally, the globules measured 0.5-1.5 mu in diameter and they were usually not membrane bound. Histochemical analysis indicated that they were lipid in nature. The inclusions were separated on a sucrose density gradient and their isolation was confirmed by electron microscopy. Thin-layer chromatography of the isolated globules suggested that they consisted predominantly of triglycerides. Since it is known that the cells synthesize triglycerides at rest, it was postulated that the structures may represent the storage form for free fatty acids which may be utilized for membrane synthesis during phagocytosis.


Transport of monoamines in membranes of adrenal chromaffin granules: physiological and pharmacological aspects. The membranes of isolated bovine adrenal chromaffin granules take up catecholamines by an ATP-dependent process which fulfills many criteria of a carriermediated energy-requiring transport. This transport together with an intragranular mechanism probably explains the high storage capacity of the chromaffin granules for catecholamines. 5.2 Other amines (e.g. 5-hydroxytryptamine (5HT), octopamine) are also taken up by the membranes, but to a different degree. This probably explains why amines normally not present in storage organelles, if available in increased amounts, may accumulate within these organelles and function as false neurohumoral transmitters. 5.3. Two mechanisms of amine uptake seem to exist at the level of the granule membranes, one reserpine-sensitive (transporting, for instance, catecholamines and 5HT), and one reserpine-resistant (transporting tryptamine and partly metaraminol). Both mechanisms depend on ATP. 5.4. Chlorpromazine interferes with the reserpine-sensitive as well as the reserpine-resistant uptake of noradrenaline (NA) by granule membranes to a similar degree. Consequently the mechanism of action of neuroleptics and reserpine at the granule membrane level are probably different. 5.5. There is only an incomplete parallelism between the inhibition of NA-uptake by granule membranes in vitro and the degree of enhancement of neuronal DA turnover in vivo caused by various neuroleptics. Therefore, factors other than interference with the amine transport through the granule membranes must be co-determinant in the neuroleptic induced enhancement of DA turnover in vivo.


[Pathophysiology of the membrane function in the kidney]. The examples of tubular proteinuria, postobstructive diuresis, tubular function in terminal renal failure and the hereditary defects of tubular transport mechanisms are used to demonstrate the difficulty in elucidating defects of renal membrane transports in man. She is caused by the impossibility of applying the appropriate techniques to evaluate membrane functions in man and by the complexity of human disease. The rapidly growing knowledge of physiologic membrane functions in the kidney should however enable some progress in the field of human pathophysiology in a not too remote future.


Cyclic AMP in the CSF of patients with schizophrenia. The dopamine hypothesis of schizophrenia proposes that schizophrenia is associated with increased brain dopaminergic function. Because dopamine is thought to stimulate the production of cyclic AMP in the brain, we hypothesized that CSF cyclic AMP would be increased in schizophrenia. Cyclic AMP in the CSF was determined in 19 schizophrenic patients who had not received neuroleptic treatment in the preceding two weeks. No significant difference could be shown between CSF cyclic AMP in these patients and CSF cyclic AMP in 10 psychiatrically normal controls.


The radiology of sclerosteosis.

The radiology of sclerosteosis. Sclerosteosis is an unusual disorder in which increased skeletal density is associated with abnormalities of bony modelling. The condition is progressive and complications arise due to cranial nerve involvement. Death from raised intracranial pressure occurs in early adulthood. In order to define the range and extent of the bony abnormalities, the radiographic features of 18 affected individuals have been analysed. The recognition of characteristic changes permits diagnostic precision.


Mixed vascular deformities of the lower limbs, with particular reference to lymphography and surgical treatment. A series of patients with congenital blood and lymph anomalies of the lower limb investigated and treated at St Thomas's Hospital, London, are reviewed. They fell into three classes: (1) those in which the venous element predominated (Klippel and other syndromes), (2) those with arteriovenous shunts and (3) those with angiomas of blood or lymph vessels scattered through the limb (diffuse mixed angiomas). Most of the patients were investigated by angiography (of blood or lymph systems) as well as by plethysmography, dermal temperature measurements and other techniques in the thermal laboratory. Phlebography showed most abnormalities in the Klippel group and was useful in delineating them before operation. The importance of confirming the existence of an adequate deep venous circulation prior to ablation of abnormal superficial vessels is emphasized. Arteriography showed most abnormalities in the group with suspected arteriovenous shunts. The most commonly performed operations in this group were for control of overgrowth of the limb or for ulceration. Lymphography showed many of the Klippel group to suffer from insufficiency of the main pathways, either aplasia or hypoplasia. In addition many had vesicles, fistulas and lymph cysts. Patients in the arteriovenous shunt group had large hyperplastic lymph pathways, which were possibly either congenital or a hypertrophic response. One hundred and thirty-eight operations were performed in 46 patients for a variety of lesions and disabilities. These are reviewed. The scope and benefit of surgery in these children are greater than has been accepted in the past. Three patients required amputation of a limb. There were 5 deaths in the series, 4 of these being in the scattered angioma group and in patients in whom the deformities extended beyond the limb into the trunk.


Cancer of the large bowel in the African: a 15-year survey at Kinshasa University Hospital, Zaïre. In a 15-year survey at Kinshasa University Hospital, 954 solid cancers were recorded. Among these, 46 malignant tumours arose in the colon, rectum and anal canal, a proportion of 4-8 per cent. A review of the patients' age and clinical presentation, and pathology and aetiology of the tumours revealed that: 1. The incidence of this cancer in young Africans is higher than in Western countries. 2. The clinical presentation of this illness in young people is frequently unusual, but older African patients commonly present with classic symptoms. 3. The incidence of mucous carcinoma tends to be higher, but could be explained by the larger number of young people suffering from the condition. 4. Intestinal amoebiasis was associated with cancer in 3 cases. Its significance remains uncertain.


Warts and wart virus antibodies in patients with systemic lupus erythematosus. Warts were found in 25 out of 56 patients with definite or probable systemic lupus erythematosus (SLE) but in only 19 out of 160 control patients. Warts were particularly prevalent in elderly patients with SLE. The corticosteroid and antimalarial drugs used to treat SLE did not influence the frequency of warts. Wart-virus antibodies were found significantly less often in patients with SLE than in controls: antibodies were detected in 23 out of 51 patients and in 40 out of 54 controls. Ihe findings suggest that some deficiency in the immune mechanisms of patients with SLE predisposes them to develop warts. There was an inverse correlation among the patients with SLE between the occurrence of warts and rheumatoid factor activity. This suggests that rheumatoid factor may interfere with resistance to warts.


Non-B hepatitis in Melbourne: a serological study of hepatitis A virus infection. A study was performed to establish the value of immune adherence haemagglutination tests for antibody to hepatitis A virus in the diagnosis of non-B hepatitis. Infection with hepatitis A virus was confirmed in 14 out of 16 patients from six families and seven out of nine patients in whom the source of infection was unknown. One additional patient, who had had two episodes of jaundice, was shown to have had an attack of hepatitis A followed by an attack of hepatitis B. In patients with acute hepatitis A antibody detectable by immune adherence haemagglutination becomes detectable three or four weeks after the onset of symptoms and reaches peak levels about five weeks later.


Differences between murine C1300 neuroblastoma clones detected by rosette formation with nerve growth factor-coated sheep red blood cells. The expression of receptors for nerve growth factor (NGF) on the cell surface was assayed by rosette formation with ligand-coated sheep red blood cells (SRBC). Cell clones derived from the murine C1300 neuroblastoma and from hybrids between a neuroblastoma clone and L cell clones showed a wide variation in the capacity to form rosettes with NGF-coated SRBC. All the neuroblastoma, L cell and hybrid clones formed rosettes with phytohemagglutinin-coated SRBC and none formed rosettes with cytochrome c- or ferritin-coated SRBC or with SRBC not coated with ligand.


Neuronal activity specific to REM sleep and its relationship to breathing. The search for the neural substrate of a state of consciousness has led to the expectation that there may be neurons which discharge tonically and rhythmically during that state alone. We have now recorded in the cat the first evidence of neurons whose rhythmic discharge is consistent with the hypothesis of a tonically active neural substrate for rapid eye movement (REM) sleep. These neurons, located in the area of gigantocellular and lateral medullary tegmental fields, begin rhythmically discharging simultaneously with the cortical desynchronization at REM sleep onset and cease firing at arousal from REM; they are essentially silent at all other times. Modulations of the discharge rate correlate with the phasic events of REM sleep, such as ataxic breathing and eye movement bursts. In addition, there is a high correlation between their discharge rate and respiratory frequency analyzed on a breath-by-breath basis. The significant rhythmicity of their discharge coupled with high REM selectivity contrasts them with putative REM generators reported by others in the pons and suggests their crucial role in REM generation.


Absence of morphine antagonism of prostaglandin E1-stimulated [3H]3',5'-cyclic adenosine monophosphate accumulation in a rat brain mince system. Prostaglandin E1 (PGE1)-stimulated [3H]3',5'-cyclic adenosine monophosphate ([3H]cAMP) accumulation was studied in a rat brain mince system to determine if stimulation of net [3H]cAMP accumulation could be prevented or reversed by morphine. Incubated minces of whole brain minus cerebellum were prepared from naive, acutely morphine-treated, and morphine-tolerant and withdrawing rats. Basal levels of [3H]cAMP accumulation in the three groups were equal within experimental limits. Morphine addition to the PGE1-stimulated minces did not prevent or reverse stimulation of [3H]cAMP accumulation in any of the three experimental groups. The data do not support the thesis that PGE1-stimulated cAMP accumulation in brain is a locus of opiate action in the phenomena of opiate analgesia, tolerance, and dependence.


Brain cyclic adenosine 3',5'-monophosphate during depolarization of the cerebral cortical cells in vivo. Cyclic AMP (cAMP) was determined in that part of the rat cerebral cortex which was invaded by slow potential change (SPC) accompanying cortical spreading depression (CSD). cAMP was determined at various phases of SPC development and at several time intervals after SPC recovery. At maximum SPC, cAMP was increased by 100% above control and by 116% at the time of SPC recovery. Five, 10 and 15 min after SPC recovery, +54%, +34% and 0% increase in cAMP was found, respectively. The activity of phosphorylase a increased by 112% at SPC recovery (maximal cAMP increase), and by 13% 15 min later (complete cAMP recovery). Some depolarizing agents which differ in their ability to stimulate cAMP formation in vitro (veratridine, cyanide and glutamate) when applied topically onto the cerebral cortex in concentrations evoking CSD (2 mM veratridine, 10 mM cyanide and 100 mM glutamate), induced uniform increase of cAMP by about 100%. The same cAMP augmentation was found in the cortical region under maximum SPC induced by these agents.


Eye movement-associated discharge in brain stem neurons during desynchronized sleep. The relationship between isolated eye movement during desynchronized sleep and unit discharge in the pontine brain stem was investigated during spontaneously occurring desynchronized sleep episodes in cats. Units histologically localized to the gigantocellular tegmental field (FTG) showed the earliest and most prominent discharge rate increases relative to eye movement onset. Inflection points for discharge rate increases of these reticular elements peaked 150-100 msec before eye movement onset and discharge maxima occurred in the time period 50 msec before and 50 msec after eye movement onset. Within the FTG there was a correlation between giant cell density and intensity of discharge rate increases relative to eye movement onset, indicating that the giant cells are the neurons that show this pattern most strongly. The exponential form of the curves of FTG inflection points and discharge maxima is compatible with self-excitation as a mechanism for recruitment in the FTG pool. Units in the central, lateral, and paralemniscal tegmental reticular fields had activity curves qualitatively similar to those of the FTG, but with less intensity of discharge rate change and shorter lead-times. The discharge pattern was quite different in the two other cell groups studied. Many units in the tegmental reticular nucleus had bursts of discharges tightly locked to eye movement onset, while pontine grey units had an irregular pattern and a low degree of phase-locking to eye movement onset. Of the neurons studied, the giant cells of the FTG best satisfy correlational criteria for eye movement generation during desynchronized sleep. These findings support the hypothesis that the FTG is part of a generator system for desynchronized sleep and may contribute directly to the elaboration of the phasic events characterizing that state.


[Necrotic enteritis in broiler chickens. III. Study of the factors favoring the multiplication of Clostridium perfringens and the experimental transmission of the disease]. Necrotic enteritis was reproduced experimentally in two week old broiler chickens by intravenous injection and also by oral administration of a pure culture of Clostridium perfringens. In the first experiment, gross and microscopic intestinal lesions, typical of necrotic enteritis, were observed in all diseased birds and mortality was obtained only in the group of birds that were injected with 0.4 ml or more of the pure culture of the microorganism. In the second experiment, the highest mortality was noted in the group of birds that received orally, in addition to the Clostridium culture, a solution of sodium bicarbonate, to obtain an alkaline intestinal content and opium to decrease the intestinal peristaltism. The gross and microscopie intestinal lesions of the diseased and killed birds were more severe than those observed in the other groups and were similar to those encountered in field outbreaks of necrotic enteritis.


Cat sleep: a unique first night effect.

Cat sleep: a unique first night effect. Longitudinal sleep studies in chronically implanted cats revealed a unique first night effect. Unlike other species, during the first 24 hr of sleep recording, the electroencephalogram and reticular formation multiple unit activity revealed more rapid eye movement sleep occurred than during subsequent 24 hr recording sessions. In addition, higher rapid eye movement to slow wave sleep ratios were observed for the first 24 hr as compared to the following days. An increase in wakefulness and decrease in slow wave sleep occurred on the third recording day as compared to the initial 24 hr period. These data have important implications for studies of variables on the sleep pattern of animals.


Interferon inducers and foot-and-mouth disease vaccines: influence of two synthetic polynucleotides on antibody response and immunity in guinea pigs and swine. Polyriboadenylic-polybouridylic acid enhanced the immunological response of guinea pigs to aqueous foot-and-mouth disease virus vaccine. Polyriboninosinic-polyribocytidylic acid enhanced the early antibody production of swine to oil emulsified foot-and-mouth disease virus vaccine. Polyriboninosinic-polyribocytidylic acid alone did not stimulate resistance to foot-and-mouth disease in swine.


Analysis of circulation of neuronal activity in the waking cortex. It is well known that the rhythmic activity of the cerebral cortex is closely associated with a highly organized circulation of neuronal impulses through the networks in which the activity develops. The present study has attempted to determine by quantitative methods, the consistency of this circulation and of its association with the cortical waves, over relatively long periods. Cortical gross waves and neuronal activity have been recorded by means of arrays of extracellular microelectrodes, in freely moving cats as well as in cats immobilized with flaxedil. Autocorrelations have been performed on trains of rhythmic waves and on associated clusters of action potentials; cross-correlations and multiple correlations have been performed between waves and action potentials and between action potentials generated by different groups of neurons. It has been found that the rhythmicity of the waves, the rhythmicity of the associated clusters of action potentials,  the time and phase relations between them, and the circulation of neuronal activity through the networks, remain highly consistent over periods as long as two hours.


Tween 80: a marker for differentiation of hog cholera and bovine viral diarrhea viruses. Markers for differentiating hog cholera and bovine viral diarrhea viruses were studied using Tween 80, chloroform, trichlorotrifluoroethane and tri (n-butyl) phosphate. Attenuated A and virulent Ames strains of hog cholera virus were employed. Moreover, the NADL PK-15 cell culture adopted strain and low cell culture passaged Purdue strain of bovine viral diarrhea virus were used. These viruses were reacted with 2,500 micrograms/ml of Tween 80 for one hour at 37 degrees C. When attenuated A and virulent Ames strains of hog cholera virus with titers greater than 10(6) and 10(5) plaque forming units respectively, were reacted with Tween 80 the titer of each strains was reduced by approximately 10(4) plaque forming units of virus. When either strain of bovine viral diarrhea virus was reacted with Tween 80, virus was not detected.


Sulfur amino acid auxotrophy in Micrococcus species isolated from human skin. Since methionine and (or) cysteine are required by a large percentage of natural auxotrophic Micrococcus strains isolated from human skin, investigations were directed to determine the specific enzymes affected in sulfur amino acid biosynthesis. Known intermediates in the interrelated cysteine and methionine biosynthetic pathways were tested as growth stimulants. Based on these growth studies, sulfur amino acid auxotrophs were grouped into three cysteine classes and five methionine classes. Selected auxotrophs of M. luteus had deficiencies in ATP sulfurylase (EC 2.7.7.4) and adenosine-5-sulfatophosphate (APS) kinase (EC 2.7.1.25), sulfite reductase (EC 1.8.1.2), serine transacetylase (EC 2.3.1.30), or beta-cystathionase (EC 4.4.1.8) activity; auxotrophs of M. lylae had deficiencies in sulfite reductase and serine transacetylase, beta-cystathionase, or N5, N10-methyltetrahydrofolate reductase (EC 1.1.1.68) activity; all auxotrophs of M. sedentarius tested had deficiencies in N5,N10-methyltetrahydrofolate reductase activity; auxotrophs of M. nishinomiyaensis had deficiencies in adenosine-3-phosphate-5-sulfatophosphate (PAPS) reductase, sulfite reductase, serine transacetylase, or N5,N10-methyltetrahydrofolate reductase activity; auxotrophs of M. varians had deficiencies in APS kinase, PAPS reductase, sulfite reductase, homoserine omicron-transsuccinylase, beta-cystathionase, or N5,N10-methyltetrahydrofolate reductase activity; auxotrophs of M. kristinae had deficiencies in serine transacetylase or cystathionine-gamma-synthase (EC 4.2.99.9) activity; auxotrophs of M. roseus had deficiencies in PAPS reductase, sulfite reductase, or serine transacetylase activity. Results of studies with various mutagens suggested that sulfur amino acid auxotrophy was primarily the result of a single base substitution in usually one or two of the genes controlling biosynthesis. A preliminary study of the amino acid composition of sweat suggested that this important source of nutrients does not contain adequate amounts of cysteine for the growth of cysteine auxotrophs but contains methionine that may be utilized in place of cysteine.


Isolation of apparently wild strains of poliovirus type 1 from sewage in the Ottawa area. In the first 4 months of 1974, 140 gauze pad samples of sewage collected in the Ottawa area were analysed by the BS-C-1 cell system for the presence of viruses pathogenic for humans. Viruses were isolated from 111 (79%) of the samples. Of the 72 (65%) isolates identified by serology and electron microscopic examination, 56 (78%) were reoviruses and 16 (22%), enteroviruses. The enterovirus isolates included one coxsackievirus B4, one vaccine strain of poliovirus type 3, nine vaccine strains of poliovirus type 1 and five strains of poliovirus type 1 that proved by serodifferentiation and temperature marker tests to be different from vaccine strains. The fact that these strains were present in the community sewage in readily detectable concentrations at a time when immunity against polioviruses is declining in such communities is a cause for concern.


Epithelioid sarcoma: ultrastructural similarity to nodular synovitis. The ultrastructure of nodular synovitis of the knee and epithelioid sarcoma of the hand are compared. Both lesions show a similar pattern of light and dark cells having filopodia and microvilli, an outer coat of finely granular matrix without well defined basal laminae, maculae adherentes and attachment sites, pinocytotic vesicles, cytoplasmic filaments and complex nuclear invaginations. These similarities suggest a common histogenesis and support the concept that the epithelioid sarcoma is derived from synovioblastic mesenchyme.


Pulmonary blastoma: an ultrastructural study with a brief review of literature and a discussion of pathogenesis. A pulmonary blastoma is reported in a 19-year-old black female. The light and electron microscopic characterics are described; they are found to be similar to those of the fetal lung prior to the fourth gestational month. Although the ultrastructural features do not favor any particular theory of histogenesis of this unique tumor, evidence for the commonly held theory that the tumor arises from pluipotential pulmonary blastema seems tenuous. A brief review of the literature disclosed 38 other similar cases. There was a male predominance with an average age of detection at 39 years. Sputum cytology was rarely positive. Among tumors larger than 5 cm in diameter, survival was usually less than 2 years. Metastases were present in approximately half of the cases.


Malignant (fibrous) histiocytoma arising in association with a bone infarct in sicle-cell disease: coincidence or cause-and-effect? A patient with homozygous-S sickle-cell disease complicated by malignant (fibrous) histiocytoma arising within a tibial medullary infarct is reported. This is the ninth reported case of human sarcoma associated with bone infarct and the first associated with sickle-cell disease. Evidence for a causal relationship is presented.


Herpetic esophagitis in patients with cancer: ante mortem diagnosis by brush cytology. Ante mortem diagnosis of herpetic esophagitis has been made in four immunosuppressed patients with cancer. The diagnosis was made by brush cytology at the time of esophagoscopy. All four patients had severe dysphagia unresponsive to nystatin therapy and the diagnosis of herpes infection excluded the use of Amphotericin B for resistant moniliasis. The diagnosis of herpes virus infection of the esophagus has been previously made almost exclusively at post mortem examination. Herpetic esophagitis is shown to be not necessarily a preterminal complication in cancer patients. In three of the four patients the esophagitis resolved as the patients responded to anti-tumor therapy.


Morphogenesis of pancreatoblastoma, infantile carcinoma of the pancreas: report of two cases. Two cases of infantile carcinoma of the pancreas were diagnosed as pancreatoblastoma based on the morphogenesis of the tumors. These encapsulated tumors adhered to the head of the pancreas and to the descending portion of the duodenum. Histologic examination revealed an organoid structure made up of cords or nodules of squamoid cells with elongated nuclei arranged in a parallel fasciculating pattern (squamoid corpuscles), surrounding tubular structures of columnar epithelial cells and intermediate light cell masses with little differentiation. Electron microscopy revealed zymogen-like granules and well developed granular endoplasmic reticulum in the cytoplasm. There were no detectable islet cells in tumor tissue. Both of these tumors could be derived from the ventral pancreas and be isolated by the lack of communication with the duct of Wirsung. As the duct of Santorini was patent, extirpation of these organoid tumors would not influence secretion of pancreatic juice. Considering the favorable prognosis after extirpation of these tumors, they should be differentiated from the usual adenocarcinoma of the pancreas occurring in adults.


Adriamycin cardiotoxicity in children: case reports, literature review, and risk factors. Two children with Wilms' tumor and prior thoracic radiation developed congestive heart failure at cumulative adriamycin doses of 350 mg/m2 and 400 mg/m2. Details of these two cases and a summary of previously reported adriamycin cardiotoxicity in children are presented. Dose recommendations and predictive tests are discussed.


Postlymphographic cerebral lipid embolization in the vena cava superior syndrome. A recent case of Lipiodol Ultrafluid embolism to the brain is reported. Pathogenetic concepts involving right-to-left shunts, and lympho-venous shunts with pulmonary capillary overloading are reviewed. In addition, it is suggested that a local disturbance in cerebral circulation due to a cava superior syndrome may contribute to the severe cerebral symptoms.


Interrelationships and functions of the pyruvate kinase isozymes and their variant forms: a review. The relationships among and the properties of the pyruvate kinase isozymes are reviewed, emphasizing their potential role in carcinogenesis. Particular consideration is given to evaluation of the concept that the three major nonreadily interconvertible forms are the products of distinct genes, the relationship of these forms to additional separable forms of pyruvate kinase, the types and possible functions of interconvertible forms of the major isozymes, and mechanisms affecting the genetic expression of the isozymes. Emphasis is placed upon the apparent derepression of the fetal isozyme in hepatomas and the influence of neoplasms and their extracts on the expression of pyruvate kinase in the liver of host animals.


Metabolism of [3H]-methylcholanthrene in the perfused rat liver. The metabolism of [3H]-3-methylcholanthrene (3-MC) was studied in the isolated, perfused rat liver. Following addition of 250 mug to the perfusion fluid, 3-MC disappeared rapidly. After 2 hr, approximately 34% of the radioactivity was excreted in the bile, 6% remained in the perfusate, and 60% was found in the liver. Of the liver radioactivity, 80% was unchanged 3-MC, 11% was conjugated metabolites, 4% was free hydroxymetabolites, and 4% was nonextractable, presumably bound to macromolecules. Of the perfusate radioactivity, 82% was conjugated metabolites, 3% was free hydroxymetabolites, and 15% was unchanged 3-MG. A similar distribution was observed in intact, bile-cannulated rats, but biliary excretion was about one-fourth as high with double the i.v.-injected dose. Biliary excretion in perfused livers rose rapidly during the first 30 to 40 min, then decreased steadily. It was nearly twice as high in male as in female rat livers. Pretreatment of rats with 3-MC more than doubled the biliary excretion rate over the first 20 to 30 min in livers of both sexes and raised that of the female to that of the male rat liver. Neither retinol acetate nor 7,8-benzoflavone had any appreciable effect on biliary excretion of 3-MC metabolites. 2-Diethylaminoethyl-2,2-diphenylvalerate, a well-known microsomal oxygenase inhibitor, lowered excretion by 80 to 90% and lengthened the lag period, and dibutyryl-3',5'-cyclic adenosine monophosphate markedly increased the rate of excretion of 3-MC metabolites. Fractionation of bile by chromatography on Sephadex LH-20 revealed six well-defined peaks of radioactivity. In contrast, bile of intact rats given 3-MC gave a pattern on Sephadex chromatography consisting of only three peaks. Preliminary data suggest that these consist of conjugates of dihydroxymetabolites as well as more highly hydroxylate derivatives. The data obtained indicate that the perfused liver is an appropriate experimental model for studies on the hepatobiliary metabolism of carcinogens.


Purine metabolic cycle in normal and leukemic leukocytes. Purine metabolism and reutilization pathways were studied as they applied to normal and leukemic leukocytes. The enzyme activities were expressed in terms of the quantity of protein extracted and per 10(10) cells. Whereas the protein extracted and the enzyme activities from normal lymphocytes were relatively constant, considerable variation was noted in cases of chronic lymphocytic leukemia (CLL). This variability in the properties of the leukemic cells suggests that the difference may be useful in the subclassification of the leukemias. The studies of the complete enzyme system were done with 300 million cells. The extraction of 350,000 normal lymphocytes/mul gave a soluble protein concentration of 1.46+/-0.16 mg protein per ml, and the yield from the same number of CLL lymphocytes varied between 0.72 and 8.32 mg protein per ml. The 5'-nucleotidase activity gave an inverse correlation with the amount of extractable protein. In individual cases of CLL, the protein concentrations and the 5'-nucleotidase activities were found on either side of the normal values. In most cases, the adenosine deaminase of CLL lymphocytic cell extracts was lower than normal, and the adenosine kinase was higher; in the CLL cells, these two enzymes gave a positive correlation with one another. Little or no difference was observed in the activities of the purine nucleoside phosphorylases in extracts of normal or leukemic lymphocytes and granulocytes. The hypoxanthine-guanine and adenine phosphoribosyltransferase activities increased in the leukemic granulocytes but almost always showed a decrease in the CLL lymphocytes when compared with the normal cells. Most of the leukemic cells had greater than normal activities of the enzymes synthesizing phosphoribosyl pyrophosphate when tested with the purines. The total nucleotide produced from adenine and guanine with adenine- and hypoxanthine-guanine phosphoribosyltransferase was about equal in normal and leukemic lymphocytes, but the proportion of the adenosine 5'-triphosphate in the product was much greater with the leukemic cells. This suggested that the ribosyltransferase activities were the same in both types of cells, but the nucleoside kinases and the nucleoside diphosphate kinases were more active in the leukemic cells. Inosine monophosphate dehydrogenase was less active than normal in the CLL cell extracts and was not directly related to the amount of inosine monophosphate generated from hypoxanthine.


Partial purification and characterization of tumor and liver S-adenosylmethionine synthetases. The S-adenosylmethionine synthetase activities of rat liver and Novikoff ascites tumor have been partially purified and characterized by chromatographic behavior, kinetic analysis, sulfhydryl dependency, and response to inhibitors. We have shown that the tumor contains a single form of the enzyme, with a Km (methionine) of 21 muM, and that the liver contains two isofunctional forms, a minor form with a Km (methionine) of 21 muM, as well as a major form with a Km of 1 mM. The tumor contained more of the low Km form of the enzyme than the liver, although the total enzyme activity of liver (measured at high substrate concentrations) exceeded that of the tumor severalfold. The tumor enzyme also corresponded to the minor form of liver enzyme in elution position from Sephadex G-150 and diethylamino-ethyl cellulose, and both had a Km (adenosine 5'-triphosphate) of 0.14 mM. The tumor enzyme differed from the major form of the liver enzymes in elution position, and the Km (adenosine 5'-triphosphate) for the latter was 1.5 mM. In contrast to the major liver enzyme, the tumor enzyme did not appear to require sulfhydryl agents for the activity to be detected, was inhibited by S-adenosylmethionine, and was inhibited to a greater degree by tripolyphosphate. It is suggested that the two forms of the enzyme are involved in the production of S-adenosylmethionine for different biological functions, and their different properties may allow selective inhibition of tumor growth by chemotherapeutic agents.


3-methylcholanthrene-induced monooxygenase (O-deethylation) activity of human lymphocytes. With a direct fluorescence assay, the levels of mixed-function oxidase activity were determined in mitogen-activated human lymphocytes. The O-deethylation of ethoxyresorufin to resorufin was used to quantitate mixed-function oxidase activity. Ethoxyresorufin O-deethylase activity was low to nondetectable in noninduced, mitogen-activated cells, but it was readily detected in 3-methylcholanthrene, mitogen-activated lymphocytes. The activity was: (a) dependent on assay time and number of lymphocytes; (b) dependent on the presence of reduced nicotinamide adenine dinucleotide phosphate; (c) stable to freezing at -80 degrees for at least 2 weeks; (d) reproducibly detected in duplicate samples of blood from one individual when cultured and assayed at the same time; but (e) quite variable in samples of blood from one individual at different times. Since in hepatic and pulmonary tissue of model animal systems ethoxyresorufin is a specific substrate for cytochrome P-448-associated monooxygenases, the use of this chemical could proffer an assay that specifically measures human cytochrome P-448-associated activity.


Metabolic regulation and relationship of endogenous protein kinase activity and steroidogenesis in isolated adrenocortical carcinoma cells of the rat. In the adrenocortical carcinoma cell, in contrast to normal isolated adrenal cells, 10 to 50 muunits of ACTH do not raise the level of adenosine cyclic 3':5'-monophosphate (cyclic AMP), protein kinase activity, and steroidogenesis. This indicates a lesion in the tumor adenylate cyclase system. Two-tenths to 10 mM cyclic AMP and guanosine cyclic 3':5'-monophosphate (cyclic GMP) which stimulate steroidogenesis in a normal cell, activate protein kinase activity in a concentration-response manner without any detectable rise in steroidogenesis in the adrenocortical carcinoma cell. Cycloheximide and actinomycin D do not inhibit the stimulation of the phosphorylation. These results suggest that the tumor cyclic nucleotide-dependent protein kinase activity is unrelated to steroidogenesis and is also not under the transcriptional or translational control steps. Curiously, muM concentrations of cyclic AMP, in contrast to cyclic GMP, stimulate protein kinase activity. In a normal cell, both cyclic AMP and cyclic GMP, in this concentration range, stimulate protein kinase without an increase in steroidogenesis. It is therefore proposed that, in contrast to the normal cell, there is an additional defect in cyclic GMP-dependent protein kinase.


Different survival of normal and transformed cells exposed to nutritional conditions nonpermissive for growth. When human diploid fibroblasts (WI-38 cells) in culture are maintained under conditions nonpermissive for growth (serum restriction), cell proliferation ceases but the cells remain viable for extended periods of time. Under similar restrictive conditions nonpermissive for growth. SV-40-transformed WI-38 fibroblasts (2RA cells) die off at a rate of 10% per day. In resting WI-38 cells, the template activity of isolated nuclei is related to both cell density and the length of time the cells have been quiescent. In 2RA cells, nuclear template activity is related to time after plating, regardless of cell density. The results are compatible with the hypothesis that normal cells can enter a G0 state more easily than transformed cells. This, in turn, would allow normal cells to survive under conditions nonpermissive for growth, whereas transformed cells gradually die off.


Mitochondrial and microsomal phospholipids of Morris hepatoma 7777. The phospholipids of both mitochondrial and microsomal membranes from normal liver, host liver, and Morris hepatoma 7777 were isolated, separated, and quantitated. The total as well as the individual fatty acid concentrations and compositions were determined. The total phosphlipids isolated from tumor mitochondria were idly altered, compared with mitochondria from other normal or host liver. The polyenoic acids were decreased, and there was a concomitant increase in the monoenes. When the respiratory control was determined, the tumor mitochondria exhibited a significant decrease in this parameter. The tumor microsomal membrane fraction, on the other hand, contained about 50% less phospholipid than the controls. The fatty acid patterns of the total as well as the individual phospholipids were quite similar to those observed in the mitochondria. The species of phosphatidylcholine from both membrane fractions were separated by argentation chromatography of the intact molecules, and, as predicted by the fatty acid compositions, the major species of the tumor was the monoenoic/dienoic fraction. The acyl coenzyme A:1-acyl glycerophosphorylcholine acyltransferases, which aid in controlling the fatty acid composition of phospholipids, were measured. The very marked increase in activity of these enzymes toward polyenoic as well as monoenic fatty acids suggested that the polyenoic acids were not available for use in the resynthesis of the phosphatidylcholines in the tumor.


Prolactin binding to R3230AC mammary carcinoma and liver in hormone-treated and diabetic rats. Specific 125I-labeled prolactin binding was measured in membrane particles prepared from R3230AC mammary carcinoma and liver of tumor-bearing Fischer rats after either prolactin, estrogen, or lergotrile mesylate treatment, or after the induction of diabetes by streptozotocin. Hormone binding to tumors was decreased by treatment with prolactin (0.5 or 1 mg/day) or estradiol valerate (7.5 mg/kg/week). In contrast, prolactin treatment did not affect prolactin binding to liver membrane particles, but estradiol valerate treatment resulted in a four-fold increase in prolactin binding to this tissue. Lergotrile mesylate, which lowers plasma prolactin levels, did not affect tumor growth or prolactin binding to either tumor or liver. Prolactin binding to both tumor and liver was significantly reduced in diabetic rats, suggesting that insulin may play an important role in controlling tissue sensitivity to prolactin. Specific binding of 125I-labeled prolactin to enzymatically dissociated cells from R3230AC tumors was demonstrated in vitro. The binding capacity of the cells was found to be of the same order of magnitude as the binding capacity in membrane preparations when appropriate corrections were applied for yields of cells and membranes. R3230AC tumor, which is responsive to prolactin appears therefore to be a useful model system for further study aimed at elucidation of growth and metabolic response to the hormone prolactin in breast cancer.


Enzyme pattern-directed chemotherapy: synergistic interaction of 3-deazauridine with D-galactosamine. We tested an experimental approach in which the specialized enzymatic pattern characteristic of the tissue of origin of a tumor might be exploited to target and enhance drug selectivity. In the present work, the D-galactosamine-induced depletion of uridine 5'-triphosphate (primarily a hepatic event) was employed to enhance the growth inhibition caused by 3-deazauridine. As predicted, the drug effect was most pronounced in the slower growing, well differentiated hepatoma lines where the activities of certain hepatic metabolic pathways and enzymes, though decreased, were still operative. The interactions of D-galactosamine and cytosine arabinoside with 3-deazauridine were examined in vitro in four liver tumor cell lines and two nonhepatic lines. The effects of D-galactosamine and 3-deazauridine on the growth of the Morris hepatoma cell lines 3924A, 8999S,AND 8999R were strongly synergistic; on the Novikoff hepatoma and the nonhepatic cell lines they were only additive. The combination of 3-deazauridine with cytosine arabinoside gave approximately additive growth inhibition with all cell types, without selective toxicity towards the hepatocellular lines. Results of growth-inhibition studies with the combination of D-galactosamine and cytosine arabinoside and with combinations of all three agents are also presented. These results are analyzed in the context of the regulation of hepatic pyrimidine nucleotide metabolism and our design of enzyme pattern directed drug selectivity.


Ca++-induced fusion of proteoliposomes: dependence on transmembrane osmotic gradient. The fusion of cytochrome oxidase liposomes with liposomes reconstituted with mitochondrial hydrophobic protein is dependent on the presence of an acidic phospholipid in the liposomes and on the addition of Ca++ions. Liposomes which have grown, by fusion, to diameters in excess of 1000 A lose the ability to fuse further, unless an osmotic gradient across the liposome membrane is established, with the internal osmotic pressure higher than the external. At a given Ca++ concentration, the extent to which this second fusion step takes place is determined by the ratio of internal to external osmolarity. Single-walled liposomes with diameters exceeding 1 mumM have been produced by this technique. The data suggest that the thermodynamic driving force for the Ca++-induced fusion is an excess surface free energy which can be supplied by membrane curvature or transmembrane osmotic gradients.


Cholestane spin label study of filipin action on lipid planar multibilayers. EPR spectra of a cholestane probe dissolved in egg yolk lecithin and lecithin-cholesterol planar multibilayers were observed as a function of the filipin dose. The probe is structurally similar to cholesterol; its normal position when dissolved is with the long axis approximately along the bilayer normal. Both cholesterol-containing and cholesterol-free samples showed spectral components characteristic of bilayer fragmentation (tilted domains) which increased with dose. Furthermore, the cholesterol-free spectra indicated that some of the probe was frozen with the long molecular axis perpendicular to the slide normal. The frozen spectral component increased with dose. Spectra from a fatty acid probe did not have this feature. We interpret this as due to probe complexed with filipin (in place of cholesterol) in accordance with the filipin-cholesterol aggregate model of deKruijff and Demel. An ultraviolet study of filipin-probe interaction indicates that the probe is capable of complexing in just such a manner but has less affinity for the drug than cholesterol. Spectra from the cholesttane probe in liposomes were also observed.


Concurrent smallpox and chickenpox.

Concurrent smallpox and chickenpox. Three patients showing smallpox- and chickenpox-like lesions simultaneously were investigated virologically. Both infections were confirmed in the laboratory and, in one case, by electron microscopy.


[High poly A content of the native genome of murine oncornaviruses : presence of 65-70 S RNA lacking polyadenylic sequences]. High molecular weight Poly (A) minus RNA was isolated from native genome of murine oncornaviruses. The Poly (A) content of viral genomes seems to depend more on cellular multiplication than viral maturation.


Two-dimensional immunoelectrophrophoretic pattern of low- and very-low-density lipoproteins, with particular reference to Fredrickson's type III. I report the pattern of low-density lipoproteins (LDL) and very-low-density lipoproteins (VLDL) as revealed by two-dimensional (crossed) immunoelectrophoresis in hyperlipoproteinemia types, II, III, and IV, preliminarily diagnosed by means of a complex lipoprotein profile examination. The higher affinity of Oil Red stain for VLDL- than for LDL-immunoprecipitation zones is peculiar to this lipoprotein class, and crossed immunoelectrophoretograms may thus be helpful for laboratory diagnosis of types IIb and IV in problematic cases. The position and the medium affinity for Oil Red of "broad beta" zones appearing in immunoelectrophoretograms of type III hyperlipoproteinemia are suggested as useful criteria for the diagnosis of this condition. From the structural and staining characteristics of VLDL-immunoprecipitation zones, alteration of both composition and antigenic properties of VLDL in hyperlipoproteinemias IIb-IV is deduced. I present in detail the procedure for crossed immunoelectrophoresis as well as that for the lipoprotein profile examination used in this study.


Interconverting measurements of human lactate dehydrogenase activities in three buffers. The lactate-to-pyruvate reaction for serum lactate dehydrogenase (LD) is most frequently assayed in one of three buffers, pyrophosphate (PPi), tris(hydroxymethyl)amino-methane (Tris), or 2-amino-2-methyl-1-propanol (AMP). We described interconverting results for serum samples and for highly purified LD isoenzymes I (dissolved in one of these matrixes) assayed in these buffers under optimized reaction conditions. The equation for converting results obtained for sera in Tris (x) to those in PPi(y) (both at 30 degrees C) is y = 0.74x+10 (n = 98). Since AMP is used extensively in Technicon procedures, we determined the LD activity of sera with an SMA 12/60, at 37 degrees C. The equation for convering these AMP results to results obtained in PPi at 30 degrees C is y = 0.45x-16 (n = 90). Very different equations were obtained with highly purified LD isoenzyme I maintained in two different matrixes and with both isoenzymes assayed in the same matrix. The matrix in which LD is dissolved and the proportion of various LD isoenzymes affect the magnitude of difference in observed LD activity under various conditions. Therefore, in clinical laboratories that use more than one analytical method or when conversion equations are used in the comparison of interlaboratory results, it is important to define the LD source, isoenzyme content, and the matrix, as well as the reaction conditions, and to use many samples with a wide range of activities when determining the conversion equations. For any changes in reagent source, substrate concentration, or alteration in procedure, a new normal range and new conversion equations should be determined.


Elevated serum and spleen angiotensin converting enzyme and serum lysozyme in Gaucher's disease. In adult chronic non-neuronopathic (Type 1) Gaucher's disease significant (p less than 0.001) elevations of angiotensin converting enzyme in serum (93.3 +/- 14.8 nmol/min/ml; number elevated, 8/11; normal control 32.2 +/- 1.30, n = 58) and spleen (5.62 +/- 0.35 nmol/min/mg protein, n = 2; control, 0.431 +/- 0.101, n = 4) and serum lysozyme (15.6 +/- 3.37 mug/ml; number elevated, 4/5) were observed. The KM for hippuryl-L-histidyl-L-leucine of Gaucher (1.31 mM) and normal (1.23 mM) serum angiotensin converting enzyme were similar. The increased angiotensin converting enzyme (ACE) in Gaucher's disease may be related to the genetic defect resulting in increased ACE synthesis in Gaucher cells, or perhaps generally, while high lysozyme may reflect an increased body mass of reticuloendothelial cells. These enzyme elevations may be of use in suggesting the possible presence of Gaucher's disease and perhaps in assessing the magnitude of pathologic involvement.


[A simple method for the separation and purification of the surface antigen of hepatitis B from serum (author's transl)]. The present report describes a simple, rapid and reproducible procedure for preparation of hepatitis B surface antigen from human positive sera, with a satisfactory yield. After removal of beta- and alpha-lipoproteins by dextran sulphate precipitation and subsequent removal of IgM and a part of the other serum proteins by dialysis against distilled water, the supernatant obtained presents almost no change in its surface antigen content. After passing this solution through a molecular sieve of Biogel A-5 m, the surface antigen is detected in a well-defined peak. The fraction correspinding to this peak is collected in its totality. After lyophilization, it is passed through a molecular sieve of CPG 10, in phosphate buffer; this allows recovery of a substance which has retained its structural as well as its antigenic properties, and which is intact and practically free of any serum contamination.


Comparison of whole plasma and perchloric acid-extracted plasma assays for carcinoembryonic antigen. A direct, paired, comparison was made of two plasma radioimmunoassays for carcinoembryonic antigen (CEA), between a perchloric acid-extracted, ammonium sulphate precipitation method; and a whole-plasma, double-antibody method. For both assays the same preparations of standard CEA, 125I-labelled CEA, and anti-CEA serum were used. Plasma samples were taken from young healthy subjects (24), cancer-free hospital inpatients (44) and cancer-proven untreated patients (94). Within-batch and between-batch variation were both greater for the extracted-plasma method. Cancer discriminatory ability was assessed by probabilistic means. With cancer-free inpatients as the reference group, no difference between the assay methods was revealed. With young healthy subjects as the reference group the whole-plasma assay was superior.


Prenatal diagnosis of galactosemia.

Prenatal diagnosis of galactosemia. Prenatal diagnosis of disorders of galactose metabolism was done in one instance in a family with a known galactokinase deficiency and in six cases in five families at risk for galactosemia. The galactokinase activity in cultured amniotic cells was found to be normal, and the diagnosis was confirmed postnatally. In the six pregnancies at risk for galactosemia, four were considered to be unaffected and two to be affected. Of the latter, one was carried to term, and the erythrocyte transferase activity of the baby was shown to be absent. The other pregnancy was terminated, and examination of fetal tissues by biochemical studies confirmed the diagnosis. This experience substantiates the concept that prenatal diagnosis of disorders of galactose metabolism is feasible.


LHRH-pituitary plasma membrane binding: the presence of specific binding sites in other tissues. Two specific binding sites for LHRH are present on plasma membranes prepared from rat and bovine anterior pituitary glands. One site is of high affinity (K = 2X108 1/MOL) and the second is of lower affinity (8-5X105 1/mol) and much greater capacity. Studies on membrane fractions prepared from other tissues showed the presence of a single specific site for LHRH. The kinetics and specificity of this site were similar to those of the lower affinity pituitary receptor. These results indicate that only pituitary membranes possess the higher affinity binding site and suggest that the low affinity site is not of physiological importance in the regulation of gonadotrophin secretion. After dissociation from membranes of non-pituitary tissues 125I-LHRH rebound to pituitary membrane preparations. Thus receptor binding per se does not result in degradation of LHRH and the function of these peripheral receptors remains obscure.


Cell-mediated immunity (CMI) to human wart virus and wart-associated tissue antigens. Lymphocyte transformation (LT) and leucocyte migration inhibition in agarose were used to demonstrate cell-mediated immune response to purified human wart virus and wart tissue extract in various subjects with warts and those without past history of warts. Most individuals bearing warts for less than 1 year duration showed positive cell-mediated responses to both the virus and tissue extract whereas very few of those who had warts for longer duration responded to either antigenic preparation. The difference was statistically significant. Subjects who had warts in the past also showed positive responses but these tended to decrease in degree with time. Surprisingly a group of subjects who never had warts before also responded to stimulation with the virus, but not to the extract. The positive response to stimulation with wart tissue extract reflects the presence of wart associated antigens other than the virus. Cell-mediated immunity against the wart virus and wart-associated antigens is probably important in preventing the persistence or even establishment of disease but this protective immunity is short-lived. The lack of quantitative correlation between LT and leucocyte migration inhibition demonstrable in this study suggests that these two are separate events in in vitro lymphocyte stimulation with antigens.


Neutrophilic granulocytes in acute bacterial infection. Sequential studies on lysozyme, myeloperoxidase and lactoferrin. The changes in intraneutrophilic and plasma concentrations of the three antibacterial proteins lysozyme, lactoferrin, and myeloperoxidase were studied sequentially during acute bacterial infection in nine patients. Intraneutrophilic concentrations of the three proteins were decreased by more than 50% during the 1st week of infection, followed by a slow increase over the following 2 weeks. Nadir values coincided with maximal toxic granulation of the neutrophils. The data suggest that neutrophilic granulocytes are deficient during early bacterial infection, possibly because of deficient synthesis of antibacterial proteins in the bone marrow, and that neutrophil toxic granulation is the visual counterpart of this defect. The plasma concentrations of the three proteins showed considerable differences: whereas plasma lysozyme did not show any sequential changes, plasma myeloperoxidase was high at the start of infection and quickly decreased towards normal values, and plasma lactoferrin, high in the first samples, showed a secondary peak 1 week after onset of disease, before normalization was seen. These differences may result from differences in the signals are specific for the individual antibacterial protein and not for the different types of neutrophil granules.


Alteration of lymphocyte subpopulations with cytomegalovirus infection in infancy. The distribution of immunoglobulin bearing (Ig+), T, and null lymphocyte subpopulations and the lymphocyte response to phytohaemagglutinin (PHA) and pokeweed mitogen (PWM) were determined in three infants with cytomegalovirus (CMV) infection. These infants had significantly decreased percentages of T cells (13%, 29% and 40%) compared to age-matched controls (61 +/- 2%). Compensatory increases in the percentages of Ig+ and null cells occurred. Decreased lymphocyte reactivity to PHA and PWM occurred in two patients. Purified T cells from these patients had normal reactivity indicating a disproportion of T cells in the peripheral blood. These abnormalities may result from CMV infection of lymphocytes and could be responsible for prolonged CMV viruria.


Abnormal 99mTechnetium-tin-pyrophosphate bone scans in chronic renal failure. A high incidence of abnormal 99mTechnetium-tin-pyrophosphate (99mTcPPi) is reported in a population of chronic renal failure patients. Using the 5 hour bone to soft tissue ratio as a quantitative index of increased uptake, 78% of 45 long-term dialysis patients and a similar proportion of non-dialyzed chronic renal failure patients were found to have increased uptakes. In animal studies using a uremic model, similar increased uptakes of 99mTcPPi was found with evidence of increased vascularity as reflected by red cell or plasma volumes in the bone or by the uptake of concomitantly administered 45Ca. The evidence suggests that the abnormal bone scans reflect abnormalities in collagen metabolism that occurs in the uremic state and that 99mTcPPi scans are useful in the diagnosis and management of renal osteodystrophy.


Tetanic fade during partial transmission failure produced by non-depolarizing neuromuscular blocking drugs in the cat. 1. A comparison has been made of the effects of three acetylcholine antagonists--hexamethonium, tubocurarine and pancuronium--on maximal tetani of limb muscles of cats under chloralose anaesthesia. In most experiments, the indirectly stimulated soleus muscle was studied, but observations were also made on the tibialis anterior and flexor digitorum longus muscles. 2. When neuromuscular block was produced by intra-arterial injections of the acetylcholine antagonists, tetanic tension, though depressed in amplitude did not wane and there was little or no post-tetanic relief of the block as judged by the amplitude of subsequent twitches. On the other hand, during similar degrees of block produced by intravenous injections, tetanic tension rapidly waned, and, after the tetanus, transmission was temporarily facilitated, as evidenced by an increase in the amplitude of post-tetanic twitches. 3. Intravenously injected hexamethonium caused complete waning of tetanic tension in doses too small to depress twitch amplitude and which caused only a small depression of peak tetanic tension. In contrast, pancuronium caused only partial tetanic fade even in doses that produced pronounced depressions of twitch and tetanic tensions. The effects of tubocurarine fell between these extremes. 4. The results suggest that depression of peak tension and tetanic fade are independent effects of acetylcholine antagonists. It is postulated that the former is a consequence of block of post-junctional cholinoceptors, whereas the latter arises from an action of pre-junctional cholinoceptors. 5. The results obtained, together with those of other workers, led to the suggestion that transmitter acetylcholine, in addition to evoking the endplate potential, acts on the nonmyelinated nerve terminals in a positive feed-back mechanism that mobilizes transmitter to keep pace with release during high frequencies of stimulation.


Effects of clonidine on vascular responses in kidney and hindlimbs of cats. 1. Cardiovascular reflexes to intravenous adrenaline and histamine and to carotid occlusion were studied in the renal vasculature, and direct effects of noradrenaline and clinidoine were compared in renal and hindlimb vascular beds in anaesthetized cats. 2. Unlike its reported effects on cardiovascular reflexes in the hindlimbs, clonidine depressed all three reflexes in the kidneys. 3. Noradrenaline caused vasoconstriction when given directly into both renal and hindlimb circulartions, but clonidine produced vasoconstriction only in the hindlimb vascular bed. 4. These results suggest that alpha-adrenoceptors in the reanl vasculature may be different from those in the hindlimbs, and that the cardiovascular reflexes in these two areas are under different control.


Altered angiotensin I conversion in pulmonary disease. 1. A specific method is described for the measurement of angiotensin I converting enzyme activity in plasma with 125 I-labelled angiotensin I used as substrate. 2. Converting enzyme activity in plasma from fifteen normal subjects, eleven patients with sarcoidosis, twelve patients with chronic obstructive pulmonary disease and three patients with shock lung was assayed by this technique. 3. Patients with sarcoidosis had increased plasma converting enzyme activity whether or not they were receiving steroid therapy. 4. Patients with chronic obstructive pulmonary disease and shock lung had decreased plasma converting enzyme activity, but extent of conversion did not correlate with the severity of the lung disease. 5. Converting enzyme activity in normal plasma could be completely inhibited by addition of exogenous angiotensin I in 0.5-2.5x107 times physiological concentration. Twice as much exogenous angiotensin I was needed to inhibit conversion completely in plasma from patients with sarcoidosis; one tenth as much in chronic obstructive pulmonary disease. These results indicate that plasma has a high capacity for angiotensin I conversion even in patients with pulmonary parenchymal disease. 6. Results suggest that plasma converting enzyme activity may be a reflection of pulmonary conversion and can be altered by pulmonary disease. 7. Measurement of plasma converting enzyme activity may be useful in studies designed to characterize the regulatory role of converting enzyme in the renin-angiotensin system and in cardiovascular homeostasis.


Isolation of bluetongue vaccine virus from infected sheep by direct inoculation onto tissue culture. A technique is described by which the vaccine strain of bluetongue virus (BTVV) may be isolated from infected fetal, neonatal, and adult sheep tissues utilizing tissue culture. The data from these studies provides evidence that 1) BTVV can be readily isolated from infected fetal and newborn tissues by tissue culture, 2) mild treatment of tissues and utilization of lysed cells as inoculum an effective means of recovering vaccine virus, 3) BTVV can be isolated with equal efficacy from mononuclear fractions and from erythrocyte granulocyte fractions of viremic blood, and 5) the brain of fetal lambs and the spleen and liver of neonatal lambs appear to be the tissues from which vaccine can be consistently isolated.


A paravaccinia virus isolated from cattle. Isolation of viruses from calves with acute respiratory tract disease were attempted on bovine embryonic lung cell cultures. An isolate obtained from one calf with oral lesions and respiratory disease, designated 44-M-E482, was characterized as a paravaccinia virus on the basis of biological and physical properties. The calf from which the paravaccinia virus 44-M-E482 was isolated did not possess serum neutralizing antibody in its convalescent sera; neither did experimentally inoculated calves possess serum neutralizing antibody to the isolate. However, a low titer of serum neutralizing antibody was produced in one calf after several intravenous injections of the virus. Inoculation of calves with 44-M-E482 into the oral mucosa, skin, nasal cavity and pharynx did not cause any noticeable illness or lesions. The relation of 44-M-E482 to the viruses which cause bovine papular stomatitis and pseudocowpox is discussed.


Cytomegaloviral infection presenting as a solitary pulmonary nodule. Cytomegaloviral infection presenting in an immunologically compromised host as a solitary pulmonary nodule has not previously been reported. A patient with a renal transplant and with no pulmonary symptoms was noted to have a single nodule on a chest roentgenogram. At autopsy, this proved to be secondary to cytomegaloviral infection. Differential diagnostic considerations in the immunosuppressed patient are discussed.


Antirival activity of N-phenyl-N'-arylthiourea derivatives against some Rhinoviruses. The effect of 12 derivatives of N-phenyl-N'-aryl- or alkylthiourea, inhibitors of human enteroviruses and foot-and-mouth disease virus, on reproduction of some rhinoviruses (H-17, B-632) in HeLa Bristol cells was studied. As screening methods both the multicylce growth test in roller tube cultures and two variants of plaque inhibition tests were employed. The compounds selected were tested in one-step growth cycle set-up. We established that N-phenyl-N'-4-hydroxyphenylthiourea (V-24) and N-phenyl-N'-2-carboxyphenylthiourea (V-17) have a distinct inhibitory effect on the growth of rhinovirus H-17, and N-phenyl-N'-2-hydroxy-5-nitrophenylthiourea (V-25) inhibited strongly the multiplication of rhinovirus B-632.


Focal dermal hypoplasia (Goltz's syndrome) manifesting as condyloma acuminatum: report of a case and review of the literature. A new case of Goltz's syndrome, focal dermal hypoplasia, is documented. The literature is reviewed. The composite individuality warrants considering Golt'z syndrome a separate syndrome in the ectodermal dysplasias. Caution against diagnosing "condyloma-like" lesions occurring in ectodermally dysplastic patients as condyloma acuminatum is urged. Neoplastic transformation and the tendency to recurrence after surgical exicision of the cutaneous angiofibromas pose a difficult problem in surgical management.


[Post-perfusion syndrome in childhood (author's transl)]. Following cardiac surgery with extracorporeal circulation three children became ill with a postperfusion syndrome. Besides typical clinical and haematological findings (fever, hepatosplenomegaly, lymphomonocytosis with "atypical" lymphocytes) demonstration of characteristic antibody sequential titres (IgG and IgM antibodies against cytomegalovirus) was possible. The importance of this syndrome rests in the problems of differential diagnosis which occur at the beginning of clinical symptoms (differentiation from endocarditis, septicaemia, hepatitis).


[Comparative studies on the removal of dental plaque by means of various dentifrices]. In order to demonstrate the cleaning effect of toothpastes which differed with regard to their percentage of polishing and wetting agents, we tested and compared different pastes ("W,X,Y,Z") to each other and with a placebo paste ("V") in 53 subjects. In a double blind trial, the Silness and Löe, Quigley and Hein plaque indices were determined, after establishing the initial values, and after two (interim measurement) and four weeks (final measurement) of application. Statistical evaluation showed that the toothpaste "w" with the components calcium phosphate, sodium laurylsulfate and sodium sulforicinoleate (in the given concentrations) cleans accessible dental surfaces from plaque and discolorations most intensively. All pastes were statistically significantly different from the control paste.


[After care of periodontally treated patients]. Follow-up examinations is mandatory with periodontal therapy in order to check the risk of recurrence. It comprises periodic check-ups, in which, at first, gingival inflammations and plaque causing such inflammation are evaluated together with the patient. Still in the same session, this is followed by conservative therapy. Apart from exact supra- and subgingival scaling, it is important to control and reinstruct the patient in personal oral hygiene. Interdental hygiene and the adjuncts selected according to the width of the inter-dental space play an important role in preventing inflammation. The time required for follow-up examinations in patients with periodontal disease calls for auxiliary personnel, especially the fully qualified dental hygienist.


Effects of methylxanthines on gonadotropin-induced steroidogenesis and protein synthesis in isolated testis interstitial cells. The effects of 1-methyl 3-isobutyl xanthine (MIX) and theophylline on basal and gonadotropin-stimulated production of cyclic AMP and testosterone were evaluated in enzyme-dispersed testicular interstitial cells. The actions of these compounds upon precursor incorporation into RNA and protein were also examined in the same cell preparation. The considerable higher potency of MIX as a phosphodiesterase inhibitor was accompanied by a steeper dose-response curve for cyclic AMP recovery in incubation media of hormone-treated cells. Both inhibitors caused increases in basal and hormone-stimulated cyclic AMP levels. Although low concentrations of theophylline and MIX had no effect on the maximum levels of hCG-stimulated testosterone production, 10 mM theophylline and 1 mM MIX significantly inhibited steroidogenesis. Conversely, basal testosterone levels were increased by MIX and theophylline. The higher concentrations of MIX and theophylline also significantly inhibited precursor incorporation into RNA and protein. These actions of phosphodiesterase inhibitors upon RNA and protein synthesis could contribute to their inhibitory effects at high concentrations upon gonadotropin-induced steroidogenesis.


Parathyroid hormone-mediated incorporation of 32P-orthophosphate into phosphatidic acid and phosphatidylinositol in renal cortical slices. Parathyroid hormone increased the incorporation of Na2H32PO4 into phosphatidic acid and phosphatidylinositol in cat renal cortical slices. Incorporation was not observed into any other phospholipid. The effects were seen as early as one minute for phosphatidic acid and ten minutes for phosphatidylinositol. 8-Bromoadnosine 3',5'-monophosphate did not mimic the effects of parathyroid hormone. Concentrations of parathyroid hormone, 1 x 10(-8)M to 1 x 10(-7)M, which increased the incorporation of 32p into phosphatidic acid and phosphatidylinositol maximally, did not alter tissue cyclic AMP levels suggesting that the incorporation of 32p was independent of cyclic AMP.


Role of 5'-guanylylimidodiphosphate in the activation of adenylyl cyclase by parathyroid hormone. We have studied the effects of guanylylimidodiphosphate (Gpp(NH)p), an analogue of GTP, on the stimulation of renal cortical adenylyl cyclase by bovine parathyroid hormone (bPTH, or bPTH-(1-84)). Incubation of canine renal membranes with bPTH-(3-34), a specific antagonist of parathyroid hormone, in either the presence or absence of Gpp(NH)p, prevented subsequently added bPTH-(1-84) from stimulating adenylyl cyclase. The addition of the antagonist to a cyclase system previously activated by both bPTH-(1-84) and Gpp(NH)p, however, produced no inhibition of enzyme activity. Removal of bPTH by washing the membranes virtually abolished activity, but washing after addition of bPTH plus Gpp(NH)p did not prevent continued accumulation of cAMP. The persistence of the activity of the enzyme brought about by the addition of Gpp(NH)p plus bPTH, despite washing or addition of specific inhibitor of bPTH action, indicates that the activity of the hormone-specific adenylyl cyclase in membrane suspensions is independent of cintinuous occupancy of the peptide-hormone receptor by bPTH in the presence of the guanyl-nucleotide analogue.


Aldosterone production by isolated glomerulosa cells: modulation of sensitivity to angiotensin II and ACTH by extracellular potassium concentration. The influence of extracellular potassium concentration on adrenal sensitivity to angiotensin II and ACTH was studied in isolated canine adrenal glomerulosa cells. When potassium was absent from the incubation medium, the aldosterone response to angiotensin II or ACTH was completely abolished. At physiologic angiotensin II concentrations (2.5 x 10(-11) M), aldosterone formation increased 4-fold when potassium concentration was increased from 2.5 to 5.0 mM, and rose 6-fold as potassium was increased from 2.5 to 7.5 mM. In the absence of angiotensin II, the same changes in potassium concentration increased aldosterone production only to 2-fold and 3.5-fold, respectively. The effect of potassium concentration upon the aldosterone response to ACTH was similar but less marked. The concentration and binding affinity of angiotensin II receptor sites in glomerulosa cells were not changed by increasing potassium concentrations from 0 to 7.5 mM. These observations demonstrate that the aldosterone response to the glomerulosa cell to angiotensin II is potassium-dependent within the physiological range for each of these stimuli. Such an interaction suggests that the in vivo effect of potassium upon aldosterone secretion includes a significant modulating action upon adrenal sensitivity to angiotensin II, as well as a direct action of potassium upon the adrenal glomerulosa cell.


On the role of the central noradrenergic and dopaminergic systems in the regulation of TSH secretion in the rat. Systemic administration of drugs affecting central noradrenergic and dopaminergic systems was used to evaluate their role in the regulation of TSH secretion in the rat. Alpha-methyl-p-tyrosine (alpha-MT) caused a depletion of brain norepinephrine and dopamine and a gradual decrease of serum TSH levels. Specific inhibitors of dopamine-beta-hydroxylase, diethyldithiocarbamate (DDC) and FLA 63, depleted central norepinephrine only and led to a simultaneous striking decrease of serum TSH. Blockade of alpha adrenergic receptors with phenoxybenzamine, but not with phentolamine, also depressed serum TSH. Blockade of beta receptors with propranolol had no effect. In contrast, the centrally and peripherally acting alpha receptor agonist, clonidine, increased serum TSH, whereas the peripherally acting methoxamine caused a decrease, probably due to non specific stress effect. A dose-related rapid inhibition of TSH secretion was observed following stimulation of dopamine receptors with apomorphine. Injection of L-Dopa had a similar effect. Blockade of the dopamine receptors with pimozide did not alter serum TSH, while blockade with spiroperidol led to a slight increase. The cold-induced surgeof TSH was abolished by pretreatment with DDC or phenoxybenzamine, reduced by apomorphine, but unaffected by pimozide or propranolol. The pituitary responsiveness to exogenous TRH was unaffected by administration of DDC or apomorphine. On the basis of these results, it is assumed that the central noradrenergic system has a stimulatory effect on the release of TRH from the hypothalamus, reflected in our experiments by the changes of serum TSH levels. It probably provides the drive for the tonic release of TRH in resting conditions and stimuli for the enhanced secretion during cold exposure. The effect is probably mediated by a central alpha-adrenergic mechanism. Activation of the dopaminergic system is inhibitory, but the physiological role of this effect remains to be established.


Regulation of casein synthesis by polyamines in mammary gland explants of mice. Studies were carried out to determine whether the actions of prolactin on the metabolism of the mammary gland may involve polyamines. In mouse mammary gland explants that were preincubated for 2 days with insulin plus hydrocortisone, the rate of [3H]leucine incorporation into casein was enhanced in a prolactin-like manner during a further incubation with spermidine plus cyclic GMP or phospholipase A. Putrescine (0.5 mM) plus PGF2alpha, cyclic GMP or arachidonic acid also enhanced the rate of casein synthesis: but PGF2alpha plus 0.5 mM arginine, ornithine or spermine had no effect. Methyl GAG, an inhibitor of the enzyme S-adenosyl-L-methionine decarboxylase (which is required for the conversion of putrescine to spermidine), abolished the putrescine plus PGF2alpha stimulation of casein synthesis. Since this drug did not affect the action of spermidine plus PGF2alpha on casein synthesis, the specific action of spermidine on casein synthesis is suggested. Neither arginine, ornithine nor the polyamines, by themselves, affected the rate of [3H]uridine incorporation into RNA or the rate of [3H]leucine incorporation into casein. Spermidine levels were elevated within 4 h after adding prolactin to explants which were preincubated for 2 days with insulin plus hydrocortisone; this effect was apparent during incubation periods of up to 48 h with prolactin. Arginase and ornithine decarboxylase activities were also elevated in response to prolactin. Arginase activity was only elevated, however, during long incubation periods with prolactin, i.e., during incubation periods of longer than 2 days. In contrast, ornithine decarboxylase activity was elevated by prolactin within a 30 min incubation period; this effect was maximal after 2 h and persisted during exposure periods of up to 24 h.