Plasma indolethylamine-n-methyltransferase activity and growth hormone level during sleep: a pilot study. Polygraphic recordings and sequential growth hormone (GH) samplings were performed in eight healthy adult males. In the plasma samples from seven of the subjects, indolethylamine-N-methyltransferase (INMT) activity was also determined. Five of eight subjects showed significant fluctuation in plasma GH level, and six of seven subjects showed significant fluctuation in plasma INMT activity level. There were also significant positive correlations between plasma GH and INMT activity level during the second episode of NREM sleep stage 1 and during the third episode of NREM sleep stage 2. A significant negative correlation between plasma GH and INMT activity level during the seventh episode of sleep stage 2 and during the fifth episode of post sleep-onset wake was found. In view of a previous finding that INMT activity in the serum of psychiatric patients is positively correlated with severity of delusions, the observation that NREM sleep is associated with mental activity characterized by repetitive thoughts, and the result that GH level in plasma is increased in NREM sleep early at night, our present findings suggest the hypotheses that increased plasma INMT activity during sleep is indicative of both increased INMT activity in the central nervous system (CNS) and the activation or maintenance of NREM mental activity during sleep. Additional research will be needed in order to validate our observations and test these hypotheses.


The effects of gamma-hydroxybutyrate on sleep. Sodium gamma-hydroxybutyrate (GHB) is a remarkably safe and nontoxic hypnotic agent which is reported to be free of addicting properties. It is also a normal metabolite of the mammalian nervous system. We examined its effects on the sleep-EEG of eight patients with histories of impaired sleep, as a prelude to a more detailed study of its clinical potential. Sleep induced with GHB was indistinguishable subjectively from natural sleep as well as by behavioral and electroencephalographic criteria. Unlike most synthetic hypnotics, GHB increased delta sleep and did not suppress REM sleep. It shortened the REM sleep latency and shifted REM sleep into the first third of the night. On one occasion it induced a sleep onset REM period which was experienced as an attack of sleep paralysis. Withdrawal was simple; there was no REM sleep rebound and sleep patterns immediately returned to their pre-drug form. Its major clinical drawback was its short duration of action: its hypnotic effect lasting only 2 to 3 hr. We suggest that GHB may serve as the prototype for a new class of hypnotic compounds derived from natural sources and capable of activating the neurological mechanisms of normal human sleep.


Erythrocyte membrane phosphorylation in hereditary spherocytosis. Phosphorylation of erythrocyte membrane proteins by membrane protein kinases was studied in 10 normal subjects and 8 patients with hereditary spherocytosis. No difference between the two groups of subjects was observed in the total phosphorylation, and, the phosphorylation of spectrin and components III and IV 5 in presence and in absence of 3',5'-cyclic AMP. So, no deficiency of membrane protein kinase activity seems to be present in hereditary spherocytosis.


[Characteristics of cerebral electrical activity in rabbits with experimental herpetic encephalitis]. Examinations carried out on 33 rabbits with an experimental herpetic encpehalitis showed changes in the electrical activity of the brain correlating with the clinical picture of the disease. A diffus slowing down of the biopotentials and paroxysmal activity of two types--the paroxysmal periodic complexes and the complexes 'spike +the paroxysmal 3/sec' was revealed during the acute period of encephalitis.


[Role of specific iodothyronine-binding proteins in the process of nuclear-cytoplasmic thyroid hormone interrelationships in liver cells]. A study was made of the role played by specific iodinethyrodine-binding proteins of rat hepatocytes in the process of nucleo-cytoplasmic interrelations of the thyroid hormones under normal conditions in thyroidectomy and in experimental thyrotoxicosis. The concentration of the hormone-binding sites in the cell depended on the thyroid hormone level outside the cell. An important role of the cytosol hormone-binding proteins in the accumulation and interacellular distribution of the thyroid cell hormones is emphasized. The absence of any participation of cytosol triiodine-thyronine binding proteins in the process ofpenetration of the hormone into the nucleus was revealed. Triiodine-thyronine level in the nuclei depended directly on the concentration of the recptor proteins in them and the degree of occupation of the acceptor sites in the receptors proper.


[Mechanism of action of ouabain and cyclic-3',5'-adenosine monophosphate on calcium ion transport in rat cardiac mitochondria]. Ouabain and cyclic AMP action on the Ca2+-accumulating capacity and the efflux of Ca2+ions from the loaded mitochondria of the rat heart was investigated by the tetracycline probe method. On the basis of the experimental results a conclusion was drawn that ouabain failed to act on these processes. Cyclic AMP didn't act on the Ca binding by the mitochondrial membrane; at the same time it induced rapid release of calcium from the loaded mitochondria.


[Fluorimetric determination of the concentration of histamine and serotonin in the mast cells of ordinary and germ-free rats following administration of reserpine and cyclic adenosine-3',5'-monophosphate]. The microspectrometric method was applied to the determination of the content of histamine (a test with orthophthalic aldehyde) and serotonin (Falk's test) in the mast cells of the mesentery of common and germfree rats. Reserpine (10 mg/kg of body weight, intramuscularly) failed to alter histamine content, whereas serotonin content fell by 50%. Administration of cyclic AMP (3.5 mg/kg of body weight, intraperitoneally) led to the increase in serotonin content by 80%, and histamine--by 280--320%.


[Effect of cyclic nucleotides and isopropylnoradrenaline on oxygen tension and absorption]. Isopropylnoradrenaline (ISO), 3',5'-AMP and dibutyryl-3',5'-AMP decreased the oxygen tension (pO2) in the liver and the spleen and increased the body oxygen consumption (VO2). Time dynamics of these two effects was closely correlated for ISO and 3',5'-AMP. An increase of heat output was not accompanied by any significant changes in the respiration coefficient. Pempidine and dihydroergotamine failed to prevent 3',5'-AMP effects; inderal somewhat decreased these effects. Apparently, the catecholamine influence upon pO2 was a result of the VO2 increase through 3'5'-AMP effects are largely direct, but they include the in vivo and beta-receptor component; 2',3'-AMP decreased pO2 and VO2.


[Effect of etimizol on energy metabolism in rat brain]. Intraperitoneal injection of ethimizol in a dose of 25 mg/kg caused in intensification of the oxidative phosphorylion, an increase in creatine phosphate and a reduction of inorganic phosphorus concentration in the tissue of the rat brain. It is supposed that stimulation of the energy metabolism by ethimizol was caused by its activating effect on adenyylcyclase.


[Rate of progress of the 1st post-stimulation division of the mitotic cycle by cells of ascitic hepatoma 22A of different ages]. A study was made of the progress rate of cells of the ascitic hepatoma 22A of different age during the iirst mitotic cycle after the stimulation of division. The "ageing" (11-day), terminal (14-day), and "delayed" (4 days older than the terminal stage) ascitic fluids were used. The maximal values of the labeled nuclei index was found to be reached by 9--12 hours (it was mainly due to the transtion of the quiescent to the S-period) and the maximal mitotic index--by 18--21 hours after the inoculation, independently of the tumour age. These results suggest that the duration of both the prereplicative (G1) period and of the whole first mitotic cycle after the stimulation were independent of the time during which the cells of the ascitic hepatoma 22A were at the resting stage or at the very prolonged G1-period.


[Role of adrenergic structures of the central nervous system in altering the function of the hypothalamo-hypophyseo-adrenal system in exogenous hyperthermia]. Subcutaneous injection of chlorpromazine to rats caused a lesser increase in body temperature and a higher content of the corticotropin-releasing factor (CRF) in the hypothalamus and the ACTH in the hypophysis under conditions of exogenous hyperthermia. The ascorbic acid content in the adrenal glands decreased in the same way as in the animals overheated without any chlorpromazine. There was no significant change in the weight of the adrenal gland. A conclusion was drawn on the participation of the central adrenergic structure in the regulation of the synthesis or the secretion of the CRF.


[Change in the resistance of partially hepatectomized mice to transplanted tumors in the period of liver regeneration completion]. There were two inhibition periods of the growth of ascitic hepatoma 22a when its cells were transplanted intraperitoneally to partially hepatectomized A/he mice 1 to 12 days after the operation. The first inhibition took place when the transplantation was performed one day, and the second--3 to 10 days, after partial hepatectomy. The animals which were most resistant to this tumour (5 to 8 days after the operation) proved to be resistant, although to a lesser extent, to the intraperitoneal transplantation of sarcoma 37 and Ehrlich's adenocarcinoma.


[Binding of 2-acetylaminofluorene-9-14C with rat liver nucleic acids during malignization and in primary hepatomas]. The residual binding of 9-14C-2-Acetylaminofluorene (9-14C-2-AAF) with rat liver nuclei acids was investigated during hepatocarcinogenesis two weeks after a single injection of 9-14C-2-AAF. Up to 6 months feeding of the animals with unlabeled 2-AAF, the RNA of their liver proved to bind increased amounts of 9-14C-2-AAF in comparison with normal liver. The binding of 9-14C-2-AAF with DNA in primary hepatomas was mainly due to the RNA heterodispersed components with the maximum level in the 18S-fraction, as well as with the biopolymere fractions with the sedimentation constant of 10 and 5S enriched with polyadenylate fragments.


[Role of macrophage destruction products in the alveolar phagocytosis reaction]. Peritoneal macrophages (PM) of Wistar rats harvested after the intraperitoneal injection of paraffin oil were destroyed by repeated freezing-thawing. When injected intratracheally to control rats or to those after 4 daily exposured to TiO2 dust, these macrophage destruction products (MDP) caused a significant rise of both the alveolar macrophages (AM) and the neutrophilic leukocytes (NL) counts in the pulmonary washing-outs; the mean NL/AM ratio increased several times as compared to rats injected with normal saline intratracheally. Thus, the response to the inert dust particles plus the exogenous MDP became similar to the one observed after the cytotoxic (for instance silica) particles inhalation. Enhancing the NL contribution to the inhaled particles phagocytosis, the MDP led to a significant decrease of the mean "Dust load" of a single AM, although the total number of the engulfed particles increased. The predominant attraction of granulocytes and particularly of the NL as compared to the peritoneal macrophages was also found in the peritoneal exudates of rats injected with the MDP or silica suspension intraperitoneally, while the alveolar phagocytosis was not influenced. In vitro the MDP was shown to stimulate the NL migration and to facilitate the O2 consumption by PM. A possible role of the MDP as a multipotent controlling factor of phagocytosis response is briefly discussed.


[Cell cultures obtained using collalytin from the pancreases of cattle fetuses]. The authors present morphological characteristics of primary monolayer cultures prepared from the pancreas of bovine fetuses. Combined treatment with trypsin and collalytine solutions (a preparation with collagenase activity) was used for dispersion of the tissue of the pancreas. Numerous epithelial cells corresponding by morphofunctional characteristics to beta-cells of the islets of Langerhans were contained in be cultures obtained; an aldehyde-fuchsin-positive granularity was revealed in the cytoplasm of these cells. Degranulation of these cells occurred under the effect of an increased glucose concentration in the nutrient medium.


[Relationship between a state of sleep and wakefulness and transmission of an afferent signal through the posterior ventral nucleus of the thalamus]. Chronic experiments were carried out on nonanesthetized cats. A study was made of peculiarities attending the transmission of the afferent signals and the following inhibitory processes in the posterior ventral nucleus of the thalamus, depending on the state of sleep and wakefulness. Transmission of the signals decreases during the dreaming condition and the slow-wave sleep, and increases during the active states of alert cats. In paradoxical sleep the signal transmission is at the same level as such at resting wakeful condition. A marked transmission fluctuation is seen during the dreaming state and in the course of the first phase of sleep. The level of signal transmission is the most stable during the active states. The degree of the trance inhibitory process after the afferent signal transmission depends on the level of anesthesia, wakefulness and natural sleep. Processes following the afferent signal transmission in the posterior ventral nucleus of the thalamus show a radical difference in the barbituiate-anesthetized in comparison with nonanesthetized animals. The trace inhibition, so characteristic of anesthesia, was expressed in the dreaming state of the animal but insignificantly; in the wakeful condition it is not manifested at all.


[Depressor reactions to sural nerve stimulation in intact unanesthetized cats]. Changes in the arterial pressure, in the heart and respiratory rate evoked by the gastrocnemuis nerve stimulation were studied on conscious cats before and during intravenous injection of noradrenaline. Stimulation of the gastrocnemius nerve increased the arterial pressure, the heart and respiratory rates. The same stimulation of the nerve during hypertension caused by noradrenaline injection led to the fall of arterial pressure and tachycardia. The depressor response failed to change under the effect of the beta-adrenoreceptor block and disappeared after the m-cholinoreceptor block with methylatropine. The depressor response was absent in the unanesthetized decerebrated cats. It is supposed that the depressor response of the arterial pressure depended on the strong cholinergic vasodilatation, reflexively evoked by stimulation of the motor nerve in the intact cats.


[Concentration of prostaglandins and cyclic adenosine-3',5'-monophosphate in the tissues of rats]. The content of prostaglandines (PG) and cyclic 3',5'-adenosine monphosphate (cAMP) was investigated in rat tissues by the radioisotopic method of competitive binding. Maximum quantities of both PG and cAMP were revealed in the same most actively functioning organs: the brain, incretory glands, small intestine. Fatty tissue showed minimum quantities of these substances. Results indicate a close functional relationship between the PG synthesis and adenylatecyclase activity in the body tissues.


[Changes in rat liver mitochondria under conditions of bilateral subdiaphramatic vagotomy]. A study was made of the changes in the mitochondria of the rat liver under conditions of bilateral subphrenic vagotomy. Two stages in the dynamics of the response of the mitochondrial apparatus to denervation were distingished. During the first stage (0.5-3 days after vagotomy) there occurred reversible functional disturbances of the mitochondria caused by the postoperative stress. The second stage (7 to 60 days after the denervation) was charaterized by more marked structural-functional changes having a number od common features with those seen in hypoxia and being result of vagotomy proper.


[Antiviral activity of preparations of RNA isolated from cells treated with interferon (messenger RNA for an antiviral protein?)]. Chick embryo cultures treated with interferon yielded a biologically active RNA which, upon inoculation into chick embryo cells, created an antiviral condition in them. The level of vesicular stomatitis virus reproduction in such cells was  2-30% of that observed in the cells treated with control RNA. The maximum activity of the experimental RNA was seen 3 hours after the treatment with interferon.


[Electron-cytochemical and morphometric study of the activity of several enzymes in thyrocyte metochondria during malignant degeneration]. Cytochrome oxidase and succinic dehydrogenase activities were studied in the mitochondria of rat thyroid cell during experimental malignization by means of the electron-cytochemical and morphometric methods. The activity of these enzymes in the mitochondria changed depending on the stage of malignization: at the early stages it approached the normal activity, and at the later stages (precancer)--it decreased and approached the mitochondrial activity of cancer cells. A sharp decrease in the activity of the enzymes under study in the morphologically changed mitochondria of cancer cells may characterize their qualitative changes.


Cyclic 3',5'-adenosine monophosphate level and adenylate cyclase activity in human blood platelets during storage in ACD solution. The level of cyclic 3',5'-adenosine monophosphate (cAMP) in human platelets and the activity of platelet adenylate cyclase in response to prostaglandin E1 stimulation do not change during two days storage at room temperature in ACD solution. However, the level of cyclic AMP is lower in platelets stored in ACD solution than in platelets from blood anticoagulated by ethylenediamine tetra-acetic acid.


Profile of beta-adrenoceptors in femoral, superior mesenteric and renal vascular beds of dogs. 1. The homogeneity of beta-adrenoceptors in femoral, superior mesenteric and renal vascular beds was investigated by the use of the regional perfusion technique in dogs. 2. Isoprenaline and salbutamol produced dose-related increases in femoral and superior mesenteric blood flow. The dose-response curves for the two agonists were parallel, but salbutamol was approximately 1/15 as potent as isoprenaline on a weight basis. 3. Isoprenaline and salbutamol increased renal blood flow in a dose-related manner. However, salbutamol was approximately 1/240 as potent as isoprenaline on a weight basis, and the slope of the dose-response curve for salbutamol was less steep than that for isoprenaline. 4. The dose-response curves to isoprenaline for increase in femoral and superior mesenteric blood flow were shifted to the right by intravenous pindolol but not by intravenous or intra-arterial practolol. 5. The dose-response curves to isoprenaline for increase in renal blood flow were shifted to the right more markedly by intravenous pindolol than by intravenous or intra-arterial practolol. 6. The results indicate that beta-adrenoceptors of the renal vascular bed consist of beta1-type and beta2-type whereas the femoral and superior mesenteric vascular beds contain only beta2-adrenoceptors.


Conjugated equine oestrogens and blood clotting: a follow-up report. A follow-up study of blood clotting and platelet aggregation was performed on 21 women who had received long-term hormone replacement treatment with conjugated equine oestrogens. The prothrombin time and factor VII and X values were significantly accelerated after three months, but there was no further increase with continual administration for 18 months. After 12 to 18 months' treatment, however, thrombin-induced platelet aggregation (Chandler's tube) was also significantly accelerated, which suggested a widening spectrum of effect. No overall acceleration of "intrinsic" clotting (partial thromboplastin time and thromboelastography) was found during the study, but the relatively small numbers may have been responsible. Further efforts are therefore required to find formulations and doses of oestrogens which, while relieving menopausal symptoms, cause less acceleration of blood clotting and platelet aggregation.


Plasma immunoreactive corticotrophin and lipotrophin in Cushing's syndrome and Addison's disease. Plasma immunoreactive corticotrophin (ACTH) and lipotrophin (LPH) were measured in patients with raised circulating concentrations from a pituitary or an ectopic source. They were measured again in seven patients after they had received hydrocortisone. Plasma ACTH concentrations were higher than LPH concentrations in patients with a pituitary source of their hormones, whereas this relation was reversed when the source was ectopic. After hydrocortisone administration the half life of immunoreactive ACTH was 40 minutes and that of LPH 95 minutes, resulting in a reversal of the normal relation of ACTH to LPH. The use of two antisera with different specificities for measuring LPH has further shown that pituitary LPH differs from ectopic LPH. Relatively less gamma-LPH than beta-LPH was produced from ectopic sources, the relation being reversed in patients with a pituitary source for their raised concentrations. Measuring plasma LPH as well as ACTH might therefore help in deciding whether a patient with Cushing's syndrome has a pituitary or ectopic source of ACTH, which sometimes presents a difficult clinical problem.


Longitudinal fibre splitting in neurogenic muscular disorders--its relation to the pathogenesis of "myopathic" change. In 15 patients with neurogenic muscular disorders, including cases of motor neuron disease, Wohlfart-Kugelberg-Welander disease, Davidenkow's scapuloperoneal syndrome, peripheral neuropathy and traumatic neuropathies, muscle biopsies were carried out, usually after EMG or single fibre EMG investigation. Enzyme histochemical and electronmicroscopic techniques were used to study longitudinal fibre splitting and its quantitative relation to the general changes in the biopsies. In 9 cases serial sections were used to study the longitudinal extent and character of fibre splitting. Longitudinal fibre splitting was found in 14 cases. It was prominent in Type 1 fibres, and in those biopsies in which hypertrophy was most marked. It was often associated with central migration of sarcolemmal nuclei. Ultrastructurally there was evidence that splitting consisted of mechanical disruption of the myofibrillar pattern, followed by an active process of membrane formation. We suggest that longitudinal splitting of muscle fibres, induced by overload of poorly innervated, hypertrophied fibres, can account for many of the "myopathic" changes found in neurogenic muscular disorders.


The neurological complications of cardiac transplantation. Review of the neurological complications encountered in 83 patients who received cardiac homografts over a seven-year period leads to the following conclusions: (1) Neurological disorders are common in transplant recipients, occurring in over 50 per cent of patients. (2) Infection was the single most frequent cause of the neurological dysfunction, being responsible for one-third of all CNS complications. (3) The infective organisms were typically those considered to be usually of low pathogenicity: fungi, viruses, protozoa and an uncommon bacterial strain. (4) Other clinical neurological syndromes were related to vascular lesions, often apparently from cerebral ischaemia or infarction occurring during the surgical procedure, metabolic encephalopathies, cerebral microglioma, acute psychotic episodes and back pain from vertebral compression fractures. (5) The infectious complications and probably the development of neoplasms de novo, are related to immunosuppressive therapy which impairs virtually all host defence mechanisms and alters the nature of the host's response to infective agents or other foreign antigens. (6) Because neurological symptoms and signs were usually those of behavioural changes or deterioration in intellectual performance, the neurological examination was often of little value in diagnosing the nature or even the anatomical site of the neuropathological process. (7) The possibility of an infectious origin of the neurological manifestations must be aggressively pursued even in the absence of fever and a significantly abnormal spinal fluid examination. The diagnostic error made most frequently was to ascribe neurological symptoms erroneously to metabolic disturbances or to "intensive care unit psychosis" when they were in fact due to unrecognized CNS infection. (8) Maintenance of mean cardiopulmonary bypass pressures above 70 mmHg, particularly in patients with known arteriosclerosis, may reduce operative morbidity. (9) Though increased diagnostic accuracy is possible with routine use of a variety of radiological and laboratory techniques, two further requirements probably must be met before a significant reduction in the frequency of neurological complications will occur: the advent of greater immunospecificity in suppressing rejection of the grafted organ while preserving defences against infection; and a more effective armamentarium of antiviral and antifungal drugs.


Effects of locus coeruleus lesions upon cerebral monoamine content, sleep-wakefulness states and the response to amphetamine in the cat. The purpose of the present study was to investigate the effects of complete lesions of the noradrenaline locus coeruleus neurons upon wakefulness and paradoxical sleep. Radiofrequency lesions of the nucleus were performed in 8 chronically implanted cats which were continuously recorded with an EEG for 5 days prior to and 21 days following the lesions, when they were sacrificed. In 3 of these animals amphetamine (2 mg/kg) was administered on one control day and on the 10th day post-lesion. Following sacrifice, monoamine content was assayed in discrete brain regions, and the lesion was examined in Nissl-stained sections of the pons. (1) The majority (x 69%) of the locus coeruleus was bilaterally destroyed by the lesions which only minimally exceeded the boundaries of the nucleus within the dorsolateral pontine tegmentum. Noradrenaline was depleted by a mean of 85% in the paleo- and neocortex and by a mean of 60% in the thalamus and midbrain. (2) EEG activation reappeared within 12-48 h following the lesion and represented a normal percentage of recording time on the 3rd and subsequent days post-lesion. The behavioral arousal and long-lasting EEG activation produced by amphetamine was qualitatively and quantitatively the same pre- and post-lesion. (3) Despite alteration of certain components, paradoxical sleep reappeared within 48 h and recovered to normal amounts by the second week post-lesion. Muscle atonia was permanently absent in 7 animals. Ponto-geniculo-occipital (PGO) spiking was acutely redistributed across all states and chronically reduced in frequency (by a mean of 50%) within paradoxical sleep. These results indicate that the noradrenaline locus coeruleus neurons are not necessary for the tonic maintenance of EEG activation that occurs in normal wakefulness and in amphetamine-produced arousal. Furthermore, these neurons are not necessary for the occurrence of paradoxical sleep, although they may be involved in modulation of PGO spiking.


Temporal patterns, their distribution and redundancy in trains of spontaneous neuronal spike intervals of the feline hippocampus studied with a non-parametric technique. A modification of the non-parametric technique for the analysis of temporal patterns in long trains of single neuronal spike intervals has been described and tested empiracally. The technique is based on inequality testing of sequential pairs of intervals. If the second interval in a pair is longer or shorter than or equal to the first interval, a(+), a(-), and a (0) is recorded respectively in sequential bins of the computer memory. Subsequently, the long sequences of signs are arranged into transition frequency matrices which are then converted into transition probability matrices of various complexity. In this manner, the sign permutations composed of 4, 5, 6, etc. signs were studied. First of all, the theoretical distribution of various sign permutations was derived, assuming that the arrangement of intervals that generate the signs is totally independent. The theoretical distribution of signs permutations in tetragrams, pentagrams and hexagrams constitute the 'controls' with which the empirical data can be compared. In this manner, using the chi-square test, the total deviation of a studied neuronal output from an independent state can be quantified. The empirical data showed a consistent deviation from the theoretical distribution of sign permutations during REM sleep, as compared to slow wave sleep which was characterized by an almost perfect theoretical distribution of sign permutations. This indicates that slow wave sleep is associated with relaxation of constraints that are responsible for the emergence of specific patterns. In addition, redundancies in the occurrence of sign permutations, and the linear relationships between them, have been defined and tested empirically. The apparent discrepancies between the redundancies, based on theoretical symmetry in sign distribution and the linear redundancy that fits the empirical data, have been defined and discussed.


Ontogeny of beta-adrenergic receptors in rat cerebral cortex. The ontogeny of beta-adrenergic receptors in rat cerebral cortex has been studied using [125I]iodohydroxybenzylpindolol as a ligand in an in vitro binding assay. The concentration of beta-adrenergic receptors was very low during the first week after birth. Between days 7 and 14 there was a rapid increase in the density of receptors. Adult levels were reached by the end of the second week. The affinities of 1-isoproterenol and iodohydroxybenzylpindolol for beta-adrenergic receptors did not vary with the age of the animal. Fluoride stimulated adenylate cyclase activity in the cerebral cortex was 40% of the adult level at birth and gradually increased to maximal levels over the next two weeks. On the other hand, catecholamine stimulated cyclic-3',5'-adenosine monophosphate accumulation was barely detectable during the first week after birth, but it increased rapidly to adult levels between days 7 and 14. The results suggest that it is the development of beta-adrenergic receptors that permits the expression of catecholamine sensitive adenylate cyclase activity. Norepinephrine stores in the cerebral cortex developed slowly reaching adult levels approximately two months after birth. There is therefore little correlation between the ontogeny of presynaptic adrenergic nerve terminals and the postsynaptic development of beta-adrenergic receptors.


Cyclic AMP-stimulated phosphorylation of a high molecular weight endogenous protein substrate in sub-cellular fractions of molluscan nervous system. Cyclic AMP stimulated the incorporation of radioactive phosphate from ATP into endogenous phosphoproteins in a 20,000 X g supernatant, as well as in a 150,000 X g supernatant, prepared from Helix nervous system homogenates. No effect of cyclic AMP was observed in the respective pellet fractions. A part (but not all) of the cyclic AMP-stimulated activity in both supernatants was due to phosphorylation of an endogenous protein substrate, of apparent molecular weight 120,000 daltons as determined from its mobility in polyacrylamide gels. Little phosphorylation of 120,000 dalton material was observed in the particulate fractions, both in the presence and absence of cyclic AMP. When the 20,000 X g pellet was incubated with radioactive ATP and an exogenous calf ovary protein kinase, cyclic AMP stimulated the phosphorylation of the 120,000 dalton material. The results suggest that both membranes and cytosol from Helix nervous system contain 120,000 dalton substrates for cyclic AMP-dependent protein kinase. However, no kinase catalyzing phosphorylation of these substrates is recovered in membrane fractions following homogenization of the tissue.


Distribution of a behaviorally highly potent ACTH4-9 analog in rat brain after intraventricular administration. Distribution within the brain of a 3-fold modified ACTH4-9 analog with a remarkably potentiated behavioral activity, 4-MET (O2), 8-d-Lys, 9-Phe-ACTH4-9, was investigated. The radioactive labeled [7-3H-Phe]ACTH4-9 analog was administered intraventricularly in urethane anesthetized rats in a dose of approximately 170 ng. Total radioactivity in CSF, measured in samples drawn from the cisterna magna, decreased over the period of 0.5-4 h after injection from 51 to 2% of the injected dose. Intraventricular injection of the ACTH4-9 analog resulted in high intact peptide levels in the brain. At 2 h after injection the distribution of radioactivity over 2500 micronm and 300 micronm frontal cut brain slices was rather homogenous. Data from distribution studies over topographically defined gross brain structures indicated that the septal area, which is involved in eliciting behavioral activities of ACTH-like neuropeptides, accumulated most of the injected radioactivity per gram wet weight. The distribution profiles within the brain of the [3H]ACTH4-9 analog and [3H]Phe showed considerable differences. Uptake studies in various brain nuclei after intraventricular administration of the [3H]ACTH4-9 analog demonstrated that the greatest part of the investigated nuclei exhibited relative low or medium uptake of radioactivity. This was also true for hippocampal and thalamic nuclei, which have been suggested as effected sites of action for ACTH peptides. Very high accumulation of radioactivity occurred only in the septal nuclei, particularly the dorsal and fimbrial septal nuclei. The results indicate selective uptake of the ACTH4-9 analog in the septal area, suggesting a possible significance of this area as a site of action of ACTH neuro-peptides.


[The endogenous metabolism of Pseudomonas fluorescens in relation to the oxidation of ethanol, serine, and pyruvate]. Studies of the oxidation of ethanol, serine and pyruvate by non proliferating suspensions of P. fluorescens show that the rate of oxidation of these substrates is appreciably increased by the addition of NADH. The observed effects are interpreted on the basis of apparent oxidation quotients of NADH which exceed the theoretical values.


[The effect of previous administration of adrenaline or isoprenaline on the activity of several hypnotics in mice]. Previous administration of adrenaline (0.5 mg/kg i.p.) and isoprenaline (10 mg/kg i.p.) enhances activity of several hypnotic drugs (pentobarbital, barbital, chloral hydrate) in mice but is without effect upon hypnotic activity of ethanol. This potentialisation is blocked by previous administration of pindolol, but not by phentolamine. Administration of SKF 525 A demonstrates that metabolism of pentobarbital is modified by this enzymatic inhibitor, which is not the case for other hypnotics.


[The effect of some pharmacological agents and electroshock on the level of cyclic AMP of the total mouse brain]. Amphetamin, pentobarbital, pargyline, parachlorophenylalanine, pentetrasol and maximal electroshock all increased significantly cyclic AMP in mice whole brain conversely reserpine induced a decrease of cyclic nucleotide. All those changes were tentatively correlated toward central monoaminergic systems activation.


[A new technic to evaluate the sensitivity of the renal parathyroid hormone receptor in man]. We propose a simplified assessment of response to parathyroid hormone which could replace the traditional Ellsworth Howard test for the differential diagnosis of pseudohypoparathyroidism in man. We have demonstrated that bovine parathyroid hormone in normal and hypoparathyroid subject increases plasma cAMP except in pseudohypoparathyroid subjects. It is an efficient, reliable method without any collection of urine samples.


[Immunocytochemical study using semi-thin sections of the phenomena of early differentiation in several cell populations of the human fetal adenohypophysis]. Immunocytological investigations have been performed on semi-thin sections of human fetal pituitaries ranging in fetal age from 6 to 26 weeks. Corticotrophs can be revealed by anti-ACTH (1-24), anti-ACTH (17-39) and anti-beta MSH but not by anti-alpha MSH immunesera from the 8th week. Somatotrophs are revealed with anti-human STH from 9th week. Differentiating cells containing only alpha subunits of glycoproteic hormones are present from 8th to 12th week. At 13th week beta subunits of TSH can be revealed immunocytologically in thyreotrophs. Beta subunits of LH or FSH can be detected in same gonadotrophic cells only from 15th week.


Selection of a stable clone of the MVPK-1 fetal porcine kidney cell for assays of foot-and-month disease virus. The MVPK-1 cell line, derived from fetal porcine kidney cells, supports the replication of foot-and-mouth disease (FMD) virus. The cell line was adapted to grow in medium containing 5% bovine serum. The susceptibility of the adapted cells decreased as they aged at 37 degrees C. Various clones were isolated from the adapted cells and their growth characteristics and sustained susceptibility to FMD virus were compared. Clone 7 maintained uniform susceptibility to FMD virus over a 3-day period at 37 degrees C and proved superior to other clones in the characteristics studied. The clone has maintained satisfactory susceptibility to FMD virus through 40 subcultures. Clone 7 can replace primary bovine kidney cells for routine viral assays, but cannot detect as much FMD virus in animal specimens as primary bovine kidney, bovine thyroid, or swine kidney cells.


Hypothalamic hypopituitarism after hemiparesis. Anterior pituitary insufficiency developed over 15 years in a man who had had a sudden episode of left hemiparesis and slurred speech at the age of 45 years. At age 60 radiographic studies revealed no evidence of a sellar or suprasellar neoplasm, and endocrinologic studies, including stimulation with thyrotropin releasing hormone and luteinizing hormone releasing hormone, yielded results consistent with hypopituitarism of hypothalamic origin. Response to thyroid, adrenocortical and testosterone therapy was good.


Renal vein plasma adenosine 3',5'-cyclic monophosphate in renovascular hypertension. The concentration of plasma adenosine 3',5'-cyclic monophosphate (cyclic AMP) and plasma renin activity (PRA) were measured concomitantly in blood from both renal veins and in arterial blood in 22 hypertensive patients. In the nine patients with true renovascular hypertension the concentration of plasma cyclic AMP was greater in the venous effluent of the kidney affected by the renal artery stenosis than in that of the unaffected or less affected kidney. The arteriovenous difference in cyclic AMP concentration was less on the affected side in all but one patient. The arteriovenous differences in PRA identified the affected kidney as the source of hyper-reninemia and showed that renin release from the other kidney was suppressed. In the 13 patients with hypertension associated with but unrelated to renal artery stenosis there were no consistent patterns of cyclic AMP concentration or PRA in the venous effluent of the kidneys or of their arteriovenous differences. In renovascular hypertension the venous effluent of the kidney affected by renal artery stenosis contains not only more renin but also more cyclic AMP, owing to either increased cyclic AMP production or decreased excretion or extraction of cyclic AMP by the affected kidney.  This unilateral increase in cyclic AMP concentration may become a complementary diagnostic feature of true renovascular hypertension.


Adjuvant chemotherapy in lung cancer: review and prospects. The results of trials testing combined surgery and chemotherapy in lung cancer are reviewed. Fifteen adjuvant trials using various chemotherapeutic agents were analyzed to determine reasons for their lack of success. Current trials with adjuvant therapy in lung cancer are briefly outlined. In addition, analysis of the activity of chemotherapeutic agents in advanced lung cancer and its implications in the design of future adjuvant studies are detailed.


Intrahepatic arterial infusion of combination of mitomycin-C and 5-fluorouracil in treatment of primary and metastatic liver carcinoma. Improvement in drug response and reduction of toxicity were observed after continuous intrahepatic arterial infusion of mytomycin-C (MMC) and 5-fluorouracil (5-FU) in 15 of 26 patients with primary or metastatic carcinoma of the liver. Serum bilirubin values of 10 mg/100 ml absence of ascites, extreme cachexia and impending hepatic failure were used as the criteria for admission of these patients into the study. The patients were given MMC in a dose of 0.08 mg/kg on day 1,5-FU in a dose of 8-10 mg/kg on days 2-5, and MMC on day 6. This schedule was reinitiated on days 8 and 15 for total mean duration of 18 days. Maintenance therapy was carried out by the administration of these drugs at induction dosage alternated each week as a single 24 hourly intravenous infusion. Objective response to combination therapy was defined as decrease of at least 50% in the liver size and in the abnormal levels of serum alkaline phosphatase and glutamic oxaloacetic transaminase (SGOT), and near normal levels of serum bilirubin for a minimum period of 2 months. The duration of objective response ranged from 3-16 months with a median of 8.2 months. The median survival time for the responders was 7.2 months for patients with primary carcinoma and 9.4 months for patients with metastatic carcinoma of the liver as compared to 2 months for patients who failed to respond to the treatment. Five out of 12 patients who were refractory to MMC or 5-FU by intravenous infusion responded to the present combination drug therapy. Of four patients who died during induction therapy, three had liver failure and the fourth suffered pulmonary embolism. These studies provide evidence that combination therapy with MMC and 5-FU increases the survival time of patients with hepatic cancer, presumably due to the synergistic action of these drugs which permits the use of a low dosage schedule and has less toxic effects.


Malignant (fibrous) histiocytoma of bone--fact or fancy?. Malignant (fibrous) histiocytoma is currently defined as a malignant primary bone tumor that contains a mixture of fibrogenic cells and cells that are histologically similar but which appear to be histiocytic. In this type of histiocytoma the nuclei are often indented; cytoplasm is usually abundant and may be slightly foamy; nucleoli are often large; and multinucleated malignant cells are usually a prominent feature. Many dedifferentiated chondrosarcomas, osteosarcomas, and fibrosarcomas of bone contain areas that resemble what we regard as malignant (fibrous) histiocytoma. When the entirety of a malignant tumor of bone fits the outlined histologic pattern, the designation of malignant (fibrous) histiocytoma seems appropriate. From 158 fibrosarcomas of bone and 962 osteosarcomas of bone in our files, 35 tumors were segregated because they appeared to be properly designated as malignant (fibrous) histiocytoma. A wide age range was represented by affected patients, and a large variety of bones harbored these tumors. Approximately one-third of patients eligible for 5-year follow-up were long-term, symptom-free survivors. Four deaths occurred from the tumor after more than 5 years, and one patient had radiographic evidence of pulmonary metastasis 7 years after amputation. Radiation therapy has been curative in at least two cases. The correct designation for these tumors in the light of current knowledge is malignant tumor, consistent with malignant (fibrous) histiocytoma.


The changing histopathology of lung cancer: a review of 1682 cases. We have reviewed the histopathology of lung cancer patients seen over the past 13 years at RPMI. Assessment of this data indicates that adenocarcinoma is becoming progressively more prevalent as related to the other forms of lung cancer. Factors which in part may account for this increased prevalence are: 1) changes in criteria for reading histopathology of lung cancer, particularly since 1967; 2) the increased incidence of lung cancer among the female population who have a propensity for adenocarcinoma; and 3) occupational and environmental factors. In 1974 adenocarcinoma for the first time became the most prevalent type of lung cancer at RPMI. Whatever the reason, if our data are truly representative of a national trend, adenocarcinoma will soon become the most prevalent type of lung cancer in the United States. This fact may result in an increasing death rate since the present 18-month survival rate for adenocarcinoma is substantially less than for squamous cell carcinoma, which has in the past been the prevalent form of the disease. As the smoking habits of women more closely approximate those of men, we expect that the incidence and mortality of lung cancer will prove to be quite similar in both sexes.


Myxoid variant of malignant fibrous histiocytoma. It has been generally recognized that malignant fibrous histiocytoma (MFH) may assume a highly myxoid, hypocellular appearance. Eighty cases of malignant fibrous histiocytoma having varying degrees of myxoid change were reviewed. These tumors typically arose on the extremities (leg, 61%; arm, 21%) of adults (peak age incidence, 60-69 years). They were usually attached to fascia (31%) or involved skeletal muscle (51%) and had a mucoid or translucent appearance. The myxoid areas consisted of widely spaced spindled and pleomorphic cells embedded in a matrix of acid mucopolysaccharides. The cellular areas were indistinguishable from those of the typical pleomorphic MFH. The rate of local recurrence of these tumors was 61%, and of metastasis, 23%, but metastasis was less likely when the tumor was small, superficially located, or had a prominent myxoid component. In fact, the degree of myxoid change was inversely related to the rate of metastasis. Therefore, because of the more favorable prognosis of the myxoid variant, it seems appropriate to separate it from the usual nonmyxoid form of MFH. The myxoid variant must also be clearly distinguished from benign myxoid lesions such as myxoma or nodular fasciitis, with which it is often confused.


Diet and cancer.

Diet and cancer. Differences in food intake patterns are positively correlated with differences in the incidence of various cancers in world populations. It is postulated here that dietary factors play a vital role in the genesis of several major neoplasms, specifically gastric and colon carcinomas. In addition to the food consumed, such variables as total caloric intake, nutrtional excess or deficit, exposure to carcinogens and consumption of alcohol also increase the risk of cancer. Through recognition of these factors and subsequent long-term manipulation of diet, it should be possible to reduce cancer morbidity and mortality.


Immunodiagnostic potential of a virus-coded, tumor-associated antigen (AG-4) in cervical cancer. The central theme of this communication is the recognition of an immunodiagnostic potential in a herpes virus antigen, the molecular interrelationship of which with cervical tumor cells is described. In addition to the productive infection caused by herpes simplex virus type 2 (HSV-2) we are confronted by latency and, as suggested by recent studies, by cancer. These different types of virus-host cell interactions are discussed at the host, as well as at the cellular level. A defined level of molecular interaction between host and viral gene products must exist if the virus is to co-exist with the host, as is the case in latency and carcinogenesis. The molecular interpretations posit the presence, in the squamous cervical tumor cells, of a product of the expression of the viral genome that has immunodiagnostic potential. The antigen designated AG-4 fulfills these predictions and appears to have immunodiagnostic potential. AG-4 is present in cervical tumor biopsies, but not in normal cervical tissue. It is a structural component of the HSV-2 virion that, in tissue cultures infected with HSV-2, is synthesized preferentially under conditions that prevent the normal replication of the virus. In view of its structural nature it is most probably virus-coded. AG-4 antibody identified in complement fixation assays with antigen prepared in tissue culture, disappears following successful tumor removal and reappears during cancer recurrence. This antibody also potentially identifies those patients with cervical atypia that are at high risk of neoplastic progression. The clinical benefits of the assay are evident.


The epidemiology of the feline leukemia virus (FeLV). Clustering of cases of feline lymphosarcoma (LSA) has been observed by veterinarians for many years. In 1964 it was discovered that feline LSA was caused by an oncornavirus, the feline leukemia virus (FeLV). In 1970, a simple, indirect immunoflourescent antibody (IFA) test for FeLV was developed which enabled large numbers of cats, living in their natural (household) environments, to be tested for the virus. In one study, over 2,000 cats were tested and the results showed conclusively that FeLV is a contagious agent for cats. This finding was independently confirmed by several other investigators using different testing procedures. After discovering the contagious nature of FeLV a test and removal program was devised which successfully prevents the spread of FeLV and the development of FeLV diseases in the pet cat population. There is, at present, no evidence that FeLV infects humans living with FeLV infected cats.


Cancer induction by exogenous hormones: possible androgen-induced cancer. Since 1973, there has been an increase in the reported number of cases of liver tumors (both benign and malignant), associated with the intake of male hormones as anabolic steroids and oral contraceptives. Because these tumors are very rare in the younger age groups, and because there are approximately 50 million women taking oral contraceptives at the present time, there is a definite need for a large-scale epidemiological investigation to clarify this possible association.


Venereal factors in human cervical cancer: evidence from marital clusters. All Caucasian women in a large Eastern city who developed pathologically confirmed cervical cancer between 1950 and 1969 are being prospectively followed in an epidemiological test of the venereal hypothesis of cervical carcinogenesis. We are attempting to identify all men who were married to these probands at any time prior to the date of their cancer diagnosis. The ultimate objective is the identification of all the other wives of the proband husbands in order that their risk of cervical cancer be assessed. A random sample of control wives similar to the other wives in age, race, date and place of marriage as well as prior marital status is also being followed. To date, a total of 1,087 other wives and 659 control wives has been fully traced. Cervical cancer or carcinoma in situ was detected in 29 (2.7%) of the other wives and in seven (1.1%) of the control wives. A total of 14.0% of the other wives had either cervical cancer or a cervical cytological specimen which was other than normal. The corresponding statistic for the control wives was 8.0%. These differences in the prevalence of cervical cancer and of non-normal cervical cytology are statistically significant. In the course of this investigation so far, we have identified 29 "marital clusters" of cervical cancer in which two women married to the same man have all developed cervical neoplasms. The observed number of 29 clusters may be compared with an expected number of 11.6. This investigation, as yet incomplete, offers confirmatory evidence of the possible role of venereal factors in the pathogenesis of human cervical neoplasia. While the genital herpesvirus is the likeliest candidate, other venereal elements might also be involved.


Oral fluorouracil therapy of hepatoma.

Oral fluorouracil therapy of hepatoma. Oral fluorouracil was administered weekly to 12 consecutive patients with unresectable hepatoma. Six patients showed an objective response with a significant increase in survival duration compared to nonresponders and to untreated patients. The clinical features of these cases are discussed, along with details of therapy, and possible reasons for the encouraging results noted with this treatment regimen.


5-Flourouracil in hepatocellular carcinoma: report of twenty-one cases. Twenty-one patients with biopsy-proven hepatocellular carcinoma were treated with oral or intravenous 5-Fluorouracil according to a randomized treatment schedule. Twenty-one patients were evaluable for response and toxicity. The two groups were comparable in distribution of pretreatment characteristics. There were no objective responses in either group. This is the largest reported group of patients with hepatocellular carcinoma treated with 5-Fluorouracil. Toxicity occurred in 46% of patients treated, but was tolerable. In contrast to previous reports, we have found that weekly 5-Fluorouracil given orally or intravenously is not of significant value in the treatment of hepatocellular carcinoma.


Combination chemotherapy for small cell carcinoma of the lung. Sixty-one patients with proven small cell carcinoma of the bronchus were treated by a five-drug pulsed chemotherapy schedule involving cyclophosphamide, vincristine, Adriamycin, methotrexate and prednisolone, given at four-week intervals. The response rate was 60%. All patients also received radiotherapy (4000-5000 rad) to the primary lesion. Survival was identical whether radiotherapy was given first or between the second and third courses of chemotherapy. The survival of responding patients was significantly prolonged as compared both with the non-responders in this series and with a similar group of patients treated only by radiotherapy in this hospital in 1970. The poor results in patients resistant to chemotherapy reflect the fact that 88% of the patients had disseminated disease at the time of diagnosis. The survival of responding patients was not improved over that reported by others as a result of incorporating Adriamycin and prednisolone into the schedule, so there seems no justification for using them in future treatments based on this pattern of chemotherapy. It was possible to identify all responding patients within six weeks of initiating chemotherapy, and a delay of this duration did not affect the efficacy of radiotherapy. Radiotherapy, however, frequently interfered with the subsequent assessment of response to chemotherapy. It therefore seems obligatory to give chemotherapy before radiotherapy in order to determine which patients will benefit from subsequent drug treatment.


Malignant mixed tumors arising in salivary glands. I. Carcinomas arising in benign mixed tumors: a clinicopathologic study. Forty-seven cases of malignant mixed tumor (MMT) arising in major and minor salivary glands are presented. By definition, all these lesions contained both a benign mixed tumor (BMT) as well as a malignant neoplasm, usually a poorly differentiated carcinoma. In some cases, the carcinoma developed in a previously untreated salivary gland mass which was known to have been present for many years, others evolved in a recurrent previously resected BMT and still others originated in a previously undetected BMT. Adverse prognostic factors included: carcinomatous involvement of the resection lines, perineural invasion, metastases in lymph nodes, and origin in a major salivary gland. Lesions arising in the palate had a better prognosis as compared to major salivary gland tumors. Some patients died of locally uncontrollable tumor and others because of metastatic disease. Local recurrences of MMT were seen frequently; 18 patients (38%) had a total of 32 local recurrences. Of patients followed five years or more, seven (35%) had died of tumor, 11 (55%) had no evidence of disease, and two (10%) had died of other causes. Prolonged follow-up is required because recurrences and death from tumor may be seen many years following the diagnosis of MMT. Recommendations for therapy are made. Criteria for differentiating MMT from recurrent BMT and from cylindromatous carcinoma are discussed.


Massive infantile nephroblastomatosis: a clinical, radiological, and pathological analysis of four cases. Four cases of massive infantile nephroblastomatosis with up to three and one-half years follow-up are described. Great similarity was found clinically, radiologically and pathologically, which distinguishes this entity from true Wilms' tumor. The course of the renal lesion following treatment was monitored by sequential radiological and biopsy studies. The pathogenesis, natural history, and management are discussed.


Comparison of nuclear nonhistone phosphoproteins of rat liver and Novikoff hepatoma. As part of a continuing comparison of nuclear proteins of tumors and other tissues, 32P-labeled nuclear proteins were extracted successively with 0.15 and 0.35 m NaC1 from the nuclei of normal, regenerating, and thioacetamide-treated rat liver as well as Novikoff hepatoma 3 hr after injection of 32Pi into rats. Separation of proteins of these fractions with aqueous phenol was carried out before two-dimensional electrophoresis on polyacrylamide gels. By autoradiography many common spots were found, but four 32P-labeled protein spots, CU', C13p, C21p, and CMp, were found in the Novikoff hepatoma and not in the various liver samples studied. Two spots, B6 and B10, were found in the liver patterns and not in the tumor. Sot B33 was very dense in regenerating liver but was only a faint spot in thioacetamide-treated liver. The greater density of Spots CU', C13p, C21p, and CMp in the tumor patterns is consistent with the increased density reported earlier for spots of the C-region of a variety of tumors.


Specific non-immunoglobulin G antibodies and cell-mediated response to herpes simplex virus antigens in women with cervical carcinoma. Non-immunoglobulin G-neutralizing antibodies to herpes simplex virus (HSV) type 2 were assayed in sera adsorbed with Staphylococcus aureus Cowan I. They were present in 8% of women with normal cervical smear and in 20, 41, and 74% of women with atypia, dysplasia, and cervical carcinoma, respectively. Lymphocytes of the patients were tested for in vitro transformation by killed HSV type 1 and HSV type 2 (HSV-2), as well as by mitomycin C-treated hamster cells transformed by HSV or other viruses or not transformed. Specific stimulation by the HSV-transformed cells occurred in 2, 22, and 40% of women with normal cervical smear, dysplasia, and carcinoma of the cervix, respectively. This frequency rose to 82% during treatment with irradiation and decreased to 0% after surgery. When HSV-2 virions were used as antigens to stimulate the lymphocytes, similar differences were found between the various groups, but they were less clear-cut, since 16% of the control women had lymphocytes responding to HSV. Non-immunoglobulin G antibodies to HSV-2 were not present in blood at the same time as cell response to HSV-2-transformed cells. There was also a negative correlation between neutralizing activities of the sera and the indices of lymphocyte stimulation, indicating a regulation between humoral and cell-mediated responses.


Association of the glycolipid pattern with antigenic alterations in mouse fibroblasts transformed by murine sarcoma virus. The level of the neutral glycolipid, Galnac beta 1 leads to 4 Gal beta 1 leads to 4Glc leads to Cer (asialo GM2), in BALB/c 3T3 mouse fibroblasts transformed by Kirsten murine sarcoma virus (3T3KIMSV) was greatly increased compared to the nontransformed parental cells (3T3). This elevated chemical quantity was found to be localized on the surface of intact cells and accessible to external reagents, as detected by immunofluorescence and labeling with galactose oxidase: NaB3H4. Furthermore, immunization of rabbits with 3T3KiMSV cells but not with 3T3 cells resulted in antibody production against asialo GM2. These results demonstrate the potential usefulness of glycolipids as tumor-associated cell surface markers.


Effect of tumor promoters on the activity of cyclic adenosine 3':5'-monophosphate-dependent and -independent protein kinases from mouse epidermis. Cyclic adenosine 3'5'-monophosphate (cyclic AMP)-dependent and -independent protein kinases were detected and partially characterized in soluble extracts from mouse epidermis. Cylic AMP-dependent histone kinase activity was separated rom cyclic AMP-independent casein kinase activity by DEAE-Sephadex chromatography. The application of the tumor promoters croton oil or 12-o-tetradecanoyl-phorbol-13-acetate to mouse skin caused a rapid increase in the soluble protein extractable from the epidermis resulting in a decrease in the specific activity of both classes of protein kinase when expressed on a protein basis. No change in the activities of either the cyclic AMP-dependent or -independent enzymes was observed when expressed relative to the DNA content.


Investigation of the rate-determining microsomal reaction of cholesterol biosynthesis from lanosterol in Morris hepatomas and liver. Previously, we reported that the properties or microsomal 4-methyl sterol demethylase isolated from liver and Morris hepatomas 5123C and 7777 are grossly similar. The individual enzymic steps of this multicomponent system have now been studied, and the rate-determining step has been determined and shown to be identical for liver and these hepatomas. The rates of microsomal oxidative attacks of the 4alpha-methyl, 4alpha-hydroxymethyl, and 4-aldehydic groups are similar for microsomes prepared from rat liver and hepatoma 7777. The rates of mixed-function oxidative attack appear to increase in the order;--CH3 less than --CH2OH less than --CHO. Furthermore, the hepatic and hepatoma NAD-dependent decarboxylase, which catalyzes the reaction following the three oxidative attacks is similar in properties and velocity. The fifth step, an NADPH-dependent reduction of the 3 ketosteroid that is produced by decarboxylation, is also similar. For both tissues, the latter two reactions, under in vitro conditions, proceed at rates that exceed the initial oxidative process. Thus, for elimination of both of the 4-methyl groups of lanosterol, the 10 individual reactions catalyzed in this multicomponent system are identical in liver and hepatoma 7777 microsomes, and the rate-determining stop for both liver and hepatoma is the inital oxidative attack on the 4alpha-methyl group of cholesterol procursors. When the rate-determining reaction of both liver and hepatoma 7777 microsomes is assayed at different temperatures, the same activation energies and the same characteristic breaks in the arrhenius plots are observed. Thus, for both liver and hepatoma, both the nature and the site of rate determination in this multienzymic system must be similar. Since the microsomal enzymes of liver nad hepatoma appear to be catalytically similar and rate determination appears to be similar, too, the characteristic lact of response of tumor microsomes to treatments in vivo that alter host liver microsomal demethylation activity suggests that the insensitivity of these tumors to dietary cholesterol should not be ascribed to alterations in the catalytic proteins. Evidence in this report suggests that the postmicrosomal supernatant fraction of both liver and hepatoma contains a cytosolic protein that may participate in the regulation of the rate-determining attack of 4alpha-methyl sterol substrates. Thus, either qualitative or quantitative differences between the postmicrosomal supernatant fractions obtained from liver and heptomas may account for the observed differences in rates of cholesterol biosynthesis.


Effect of inhibiting DNA, RNA, and protein synthesis of tumor cells on their susceptibility to killing by antibody and complement. A number of metabolic inhibitors and chemotherapeutic agents have been found to increase the sensitivity of a chemically induced guinea pig hepatoma (line 1) to killing by antibody and complement. We have investigated whether the mechanism whereby these drugs increase sensitivity to killing is attributable to their primary action of inhibiting DNA, RNA, or protein synthesis. Line 1 cells incubated for 1, 4, or 17 hr with actinomycin D (25 microng/ml), adriamycin (40 microng/ml), or puromycin (5 micron/ml) or with 5-fold lower concentrations of these drugs were maximally inhibited (greater than 90%) in their ability to synthesize DNA, RNA, and protein within 1 hr. However, only cells incubated for 17 hr with the high concentrations of drugs showed increased sensitivity to killing by antibody and complement. Line 1 cells incubated with high concentrations of these drugs of 17 hr, washed, and resuspended in drug-free medium recovered their resistance to killing by antibody and complement within 4 hr. These cells ever after culture for 24 hr in drug-free medium did not regain their ability to synthesize DNA, RNA, or protein. A similar lack of correlation between synthesis of these macromolecules and sensitivity to antibody-complement-mediated killing was observed after the cells were treated with physical agents that inhibit macromolecular synthesis. Both heat-treated and X-irradiated cells were inhibited in their ability to synthesize DNA, RNA, and protein immediately after treatment; however, only X-irradiated cells (6 and 16 hr postirradiation) were increased in their sensitivity to antibody-complement-mediated killing. Our data show that the ability of line 1 tumor cells to resist humoral immune attack does not depend solely on their ability to synthesize DNA, RNA, or protein.


Procedures for radioimmunoassay of the mouse mammary tumor virus. A procedure for radioimmunoassay of the major glycoprotein antigen derived from murine mammary tumor virus is described. The assay is sensitive to 0.05 ng of antigen and is highly reproducible. The antigen, gp55, has been found to be group specific and will detect viruses in 13 separate mouse strains, as wel  .l as from continuous cell lines. Factors affecting the assay have been examined.


Cross-neutralization of ovine and bovine C-type leukemia virus-induced syncytia formation. Ovine leukemia virus (OLV) could not be distinguished from bovine leukemia virus (BLV) morphologically. Both C-type RNA tumor virus-producing cells induced syncytia when cocultivated with several indicator cells including XC and KC cells. Syncytia formation could be blocked by antibodies against OLV and BLV, indicating that fusion was virus mediated. Furthermore, antibodies against OLV and BLV exhibited reciprocal syncytial blocking properties, suggesting that OLV and BLV were closely related viruses.


Glycosyltransferases and glycosidases in Morris hepatomas. The following three parameters were studied in Morris hepatomas of different growth rates: (a) the specific activity of guanosine dephosphate (GDP)-fucose:glycoprotein fucosyltransferase and cytidine monophosphate (CMP)-N-acetylneuraminic acid:glycoprotein sialyltransferase, (B) the content of GDP-fucosee and CMP-N-acetylneuraminic acid, and (c) the activity of alpha-L-fucosidase and neuraminidase. Fucosyltrasferase activities were significantly elevated in all hepatomas investigated. Especially high levels of enzyme were measured in the rapidly growing tumors 7777, 66, and 3924A. The increase varied between 2- and 3-fold when compared with the corresponding host liver. Conversely, the activity of the sialytransferase was greatly decreased in all hepatoma lines with a rapid or intermediate growth rate. In the fast-growing tumor 9618A2, the activity was reduced to 8%. GDP-fucose and CMP-N-acetylneuraminic acid were determined by the isotope dilution technique. In normal rat liver from Buffalo or ACl rats, the concentration of GDP-fucose was 6.5+/-0.9 and 9.5+/-1.1nmoles/g, wet weight, respectively. In the fast-growing hepatomas 3924A and 9121, levels up to 21.5 nmoles/g, wet weight, were found, However, the content of CMP-N-acetylneuraminic acid in hepatomas was indluenced to a lesser extent by the degree of differentiation of the tumor. In the most rapidly growing tumor, 9618A2, a level of alpha-L- fucosidase seven times higher than in host liver was determined. Moreover, there existed a correlation bewteen the age of the hepatoma and enzyme activity. Within the 2nd week after inoculation, fucosidase activity increased from 130 to 343 nmoles/hr/mg of protein. Neuraminidase was measured in a new linked assay system. The activity of this enzyme was lowered by 50% or was at least unchanged when compared to the activity in host liver. Our results indicate that specific alterations of fucose metabolism are a characteristic feature of Morris hepatomas.


Elevated expression of T-antigen in simian papovavirus 40-infected skin fibroblasts from individuals with cytogenetic defects. A number of cytogenetic conditions were examined for expression of simian papovavirus 40 T-antigen in vitro. Skin fibroblasts from patients with Turner's syndrome and trisomy 18 syndrome and most cell lines from Klinefelter's syndrome, trisomy 13 syndrome, chromosomal translocations, chromosome 21 deletions, and single cases of 18q- and 4p- exhibited elevated T-antigen expression, compared to a clinically and cytogenetically normal control population. Thus, T-antigen expression was generally elevated in cells with increased, decreased, or rearranged genetic material involving many different chromosomes. Variation in T-antigen expression among cell lines may reflect two factors. Individual cell line factors may account for differences within homogeneous clinical groups, whereas population factors appear to account for differences between the various clinical groups and the control population. The observation of elevated T-antigen expression in diverse cytogenetic conditions suggests that this phenomenon may be a manifestation of chromosomal aberration unrelated to cancer susceptibility.


Characterization of pyrimidine nucleoside monophosphokinase in normal and malignant tissues. It was found that there are two kinds of pyrimidine nucleoside, monophosphokinase deoxythymidine 5'-monophosphate-deoxyuridine 5'-monophosphate (dTMP-dUMP) kinase and cytidine 5'-monophosphate-deoxycytidine 5'-monophosphate-uridine 5'-monophosphate-doexyuridine 5'-monophosphate (CMP-dCMP-UMP-dUMP) kinase, and their molecular weights were calculated to be 46,000 and 26,000, respectively, by gel filtration. dTMP-dUMP kinase phosphorylated dTMP with a Km of 3.1 X 10(-5)M and dUMP with a Km of 7.7 X 10(-4) M. dTMP phosphorylation catalyzed by dTMP-dUMP kinase was inhibited competively by dUMP with a Ki of 2.0 X 10(-3) M. Similarly, phosphorylation of dUMP by this enzyme was inhibited competively by dTMP with a Ki of 2.5 X 10 (-5) M. CMP-dCMP-UMP-dUMP kinase of Yoshida sarcoma phosphorylated dUMP with a Km of 3.1 X 10(-3) M and dCMP with a Km of 7.1 X 10 (-4) M, but it did not phosphorylate dTMP. Phosphorylation of dUMP BY CMP-dCMP-UMP-dUMP kinase was inhibited competitively by DCMP and dTMP with Ki's of 6.9 X 10(-4) and 3.0 X 10(-3) M, respectively, and phosphorylation of dCMP was inhibited completely by dUMP a Ki of 2.2 X 10(-3) M. Relative Vmax activity of this enzyme was 345 nmoles/mg protein with dCMP and 127 nmoles/mg protein with dUMP.


Estrogen receptors in androgen-induced breast tumor regression. The hormone-dependent 7,12-dimethylbenz(a)anthracene rat mammary tumor has been shown to regress when administered pharmacological doses of testosterone propionate. Tumor regression was correlated with estrogen receptor before and 15 to 20 days following testosterone therapy. A dramatic decline of receptor occurred in all regressing tumors, whereas those administered sesame oil alone maintained both growth and receptor content. Although receptor in regressing tumor was significantly less than in the untreated biopsies, the small amount of remaining receptor maintained the same binding affinity to estradiol, showing that testosterone affects the number and not estrogen affinity of the estrogen receptor. These studies suggest that testosterone depletion of estrogen receptor may be causally related to tumor regression.


Influence of age at inoculation on avian oncornavirus-induced brain tumor incidence, tumor morphology, and postinoculation survival in F344 rats. Intracranial neoplasms were induced by intracerebral inoculation of a standardized, cell-free inoculum of the Bratislava-77 strain of avian sarcoma virus in F344 rats at 1, 9,97 to 99, and 528 days of age. Deaths from diseases that occur spontaneously in aged F344 rats complicated assessment of tumor incidence in rats inoculated at 528 days; 20 of 30 rats inoculated at this age developed brain tumors. All rats inoculated at age 1 day (47 rats), at age 9 days (37 rats), and at 97 to 99 days of age (41 rats) developed brain tumors. The incidence of animals developing tumors was 100% in these three groups, but the incidence of multiple tumors declined with increasing age at inoculation. The mean and variance of postinoculation survival increased from 83.8 +/- 21.5 days for rats inoculated at 1 day of age to 284.6 +/- 151.5 days for rats inoculated at 97 to 99 days of age. Poorly differentiated astrocytomas and astrocytomas of mixed morphology were common among rats inoculated as neonates. Solitary, pilocytic astrocytomas were the most common tumors among rats inoculated as adults.


Comparison of the blood supply to diethylnitrosamine-induced hyperplastic nodules and hepatomas and to the surrounding liver. Intravascular injection of radionuclide-labeled microspheres was used to compare the blood supply to diethylnitrosamine-induced hyperplastic liver nodules and hepatomas with the blood supply to the surrounding, histologically normal liver. Microspheres injected into the heart or portal vein lodged in the organs of control and diethylnitrosamine-treated rats providing a quantitative index of blood supply to the microvascular bed. The blood supply is expressed as percentage of cardiac output (arterial) or cpm (portal) per organ, lobe, g tissue, etc. The fraction of the cardiac output received by lung, kidneys, spleen, and liver was similar in control and carcinogen-treated animals. The arterial blood supply of 23 nodules and hepatomas was variable [1.17 +/- 0.22% (S.E.) cardiac output per g, fixed weight], but it was similar to the arterial supply to the surrounding tissue (1.12 +/- 0.21% cardiac output per g, fixed weight). In contrast the portal blood supply to 25 selected lesions wa 39 +/- 6% that of the surrounding liver tissue. There was no apparent relationship between blood supply and lesion size or histological appearance. While only 0.13 +/- 0.04% of the microspheres injected via the portal system were recovered in the lungs of control rats, approximately 100 times this number bypassed or escaped the liver containing nodules and hepatomas and lodged in the lungs. Such alterations in blood flow could contribute to biological diversification of hepatic lesions in successive stages of cancer evolution and could facilitate metastasis from the liver.


DNA transfection of ecotropic murine leukemia viruses in mouse cell cultures. DNA's were isolated from cells chronically infected with N-, B-, or NB-tropic murine leukemia viruses and tested for infectious activity in various mouse cell cultures. Early detection of the DNA transfection is facilitated by growing the DNA-recipient cells in medium containing 10(-6) M hydrocortisone. Appropriate shearing of the DNA preparations may increase the efficiency of the transfection. With these procedures virus production of the transfected cells can be detected by XC plaque assay as early as 4 days after DNA inoculation in NIH 3T3 cells. Susceptibility of the mouse cell cultures to DNA transfection does not parallel their susceptibility to virion infection. Progeny viruses derived from the transfection show the same N- or B-tropic host range property as do the parent viruses.


Elevated concentration of a dexamethasone-receptor translocation inhibitor in Novikoff hepatoma cells. Despite the apparent similarity of the dexamethasone-receptor concentration in the rat liver and Novikoff hepatoma cytoplasms, the in vitro translocation of the dexamethasone-receptor complex into liver or hepatoma nuclei was three times greater from liver as compared to hepatoma cytosol; both cytosols showed an apparent saturation of nuclear receptor sites. Although the competitive interaction of the receptor complex with nuclei suggested that they were qualitatively similar, the translocation of additional dexamethasone-receptor from liver cytosol into nuclei presaturated with the receptor complex from hepatoma cytosol suggested that they were different. However, the latter observation, as well as the initial differential nuclear uptake of the dexamethasone-receptor complex from liver and hepatoma cytosols, can be accounted for by a higher concentration of a translocation inhibitor in the hepatoma cytosol. Thus hepatoma cytosol, at a protein concentration of 6 mg/ml and free of activated receptor complex, was four times more inhibitory to nuclear uptake of the activated complex than was a comparable preparation of liver cytosol. A study of the kinetics of nuclear uptake of the partially purified dexamethasone-receptor complex from hepatoma confirmed that, in the absence of cytoplasmic translocation inhibitor, nuclear acceptor sites were not limiting in vitro. Evidence is presented which suggests, but does not prove, that the cytoplasmic translocation inhibitor functions as such in the intact cell.


Murine sarcoma virus pseudotypes used as immunogens against viral and chemical oncogenesis. The purpose of this study was to develop an animal system of protective immunity against oncornaviruses and to test whether such immunization had an inhibitory effect upon chemical sarcomagenesis. Several murine sarcoma virus (MSV) pseudotypes were used as immunogens and tested against themselves, against other pseudotypes, against leukemogenesis by their helper viruses, and against sarcomagenesis by 3-methylcholanthrene. Five MSV pseudotypes were obtained by rescuing complete MSV from MSV-genome carrier, nonproducer hamster tumor cells, using five different leukemia viruses as helpers. The immunogenic properties of these pseudotypes could be specified on the basis of the following observations. 1) They all induced sarcomas in newborn mice and regressing sarcoma nodules in young adult mice. After regression, most mice remained free of neoplastic disease, but some developed sarcoma or leukemia relapses. 2) They had an individual host range pattern, usually determined by the helper virus, as tested by inoculation of a constant virus dose in BALB/c, C57BL/Ka, and Swiss mice. 3) They were all immunogenic, in the sense that the first virus inoculation prevented sarcoma induction by a second challenge, either viral or cellular. 4) They were cross-reactive in vivo, one pseudotype immunizing against another, in the combinations tested. 5) They were able to immunize against leukemogenesis induced by their helper viruses. This was shown by prevention of leukemic deaths by Rauscher and Friend viruses, by a slight prolongation of survival after challenge with the Precerutti-Law leukemia virus, and by inhibition of splenomegaly by Moloney leukemia virus. In a second stage of the study, we investigated whether immunization with any of the MSV psuedotypes had an inhibitory effect upon sarcomagenesis induced by near-threshold doses of 3-methylcholanthrene. The incidence of these sarcomas was essentially the same in virus-immunized and control mice. It was concluded that immunizing procedures able to prevent sarcomagenesis when the inducer is a virus did not have any consistent preventive effect when the inducer was a chemical.


Repair of DNA in human cells after treatment with activated aflatoxin B1. Repair replication was examined in cultured human cells exposed to the hepatocarcinogen aflatoxin B1 using the combined bromodeoxyuridine density label and radioisotopic label method. Semiconservative DNA synthesis was strongly inhibited, and the repair replication mode was stimulated in diploid fibroblasts (W138) and in their SV40 transformants (VA13) only when exposure to aflatoxin B1 was in the presence of an activating system containing rat liver microsomal enzymes. The maximum amount of repair synthesis was about 20% of that obtained after saturating doses of ultraviolet light (UV). The time course of repair synthesis was similar to that seen after UV, and most of the synthesis was in 30- to 50-nucleotide "short patches." A line of SV40-transformed xeroderma pigmentosum cells (Group A) deficient in repair after exposure to UV was similarly deficient in repair replication after aflatoxin treatment. Treatment with aflatoxin resulted in a 25 to 45% inhibition of UV-induced repair replication, suggesting that in addition to producing lesions in DNA, which are substrates for the excision repair system, the toxin also inhibits excision repair. CsC1 gradients of DNA treated in vitro with activated aflatoxin demonstrated binding of the drug to DNA. Alkaline sucrose gradient sedimentation gave no indication that single-strand breaks or alkali labile bonds were introduced into DNA by treatment of cells with activated aflatoxin.


Leukemogenesis in vitro induced by thymus epithelial reticulum cells transmitting murine leukemia viruses. The role of the thymus in induction of leukemia was studied in vitro. Curltivation of normal thymus cells on thymus epithelial reticulum cell monolayers that had been grown from radiation leukemia virus-induced leukemic thymuses rendered the thymocytes leukemic. C57BL/6 thymocytes were cultivated for 3 days on leukemic thymus reticulum monolayers, and 106 thymocytes were injected i.p. into young adult C57BL/6 mice. After 3 to 4 weeks all mice died of disseminated lymphatic leukemia. Mice given thymocytes that had been cultivated on thymus epithelial reticulum monolayers from normal mice did not develop lymphomas. The leukemic thymus epithelial reticulum cells were shown to produce thymotropic as well as ecotropic and xenotropic radiation leukemia virus. (Thymotropic virus has affinity for thymus lymphocytes but noes not infect fibroblasts.) The cells were brightly positive for murine leukemia virus group-specific antigen in immunofluorescence tests. Leukemic thymus epithelial reticulum cells produced ample infectious exotropic virus in the culture supernatant, although the cells were negative in the XC syncytia test. Upon infection of mouse fibroblasts with ecotropic virus produced by the leukemic reticulum cells, XC syncytia were readily obtained. Thymocytes that were cultivated on leukemic thymus reticulum cells became positive for murine leukemia virus group-specific antigen and produced syncytia in the XC test. Thus, in vitro lymphomagenesis of the thymocytes that were cultured on leukemic thymus reticulum cells was associated with their infection with thymotropic and ecotropic radiation leukemia virus.


Comparison of the growth properties in vitro and transplantability of continuous mouse mammary tumor cell lines and clonal derivatives. Five continuous mouse mammary tumor cell lines and 12 clonal derivatives have been established from tumors arising in BALB/c or C3H mice either spontaneously or in response to viral (mammary tumor virus), hormonal (17 beta-estradiol), or chemical [7,12-dimethylbenz(alpha)anthracene] stimuli in vivo. The cell lines were examined for the following in vitro growth parameters:plating efficiency, saturation density, population-doubling times, colony formation on plastic surfaces, and anchorage-independent growth in methylcellulose. The majority of the mammary tumor cell lines were transplantable in syngeneic, immunocompetent mice and gave rise to tumors composed of epithelial cells. There was no growth parameter in vitro which invariably correlated with tumorigenicity in vivo. Most of the mammary tumor cell lines appeared to produce C-type virus. Only one, a C3H derivative, appeared to respond to stimulation by glucocorticoids with the enhanced production of B-type virus. Analysis of the growth properties exhibited in culture by transformed mammary epithelial cells revealed marked differences from those previously reported for transformed fibroblasts.


Factors affecting deoxycytidylate deaminase activity in some transplantable rat hepatomas. Dunning hepatoma has a low activity of deoxycytidylate deaminase, comparable to that of normal adult rat liver. This activity seems inconsistent with the rapid proliferation rate of the tumor. Factors which might affect the activity of deoxycytidylate deaminase in the Dunning hepatoma have been examined in it and compared to the Novikoff hepatoma which has high activity of this enzyme. The low activity in Dunning hepatoma does not appear to be the result of any inhibition or, possibly, proteolytic enzyme as judged by mixing experiments, nor does it appear to be due to in vivo differences in nucleotide concentrations especially deoxycytidine 5'-monophosphate, deoxycytidine 5'-triphosphate, or deoxyguanosine 5'-monophosphate which might either help stabilize the enzyme, allosterically increase its activity, or inhibit it. The Dunning hepatoma does not convert cytosine deoxyriboside to uridine deoxyriboside at a significant rate, and the formation of uridine deoxyriboside from deoxyuridine monophosphate is 1% or less during a 30-min incubation of high-speed supernatant fraction from the tumor in either the presence or absence of fluoride. It is concluded that the Dunning hepatoma probably has intrinsically low deoxycytidylate deaminase activity.


The reduction of uridine 5'-diphosphate and uridine 5'-triphosphate in some transplantable rat hepatomas. The reduction of uridine 5'-diphosphate (UDP) and uridine 5'-triphosphate (UTP) has been studied in normal adult rat liver, the Dunning hepatoma, and Morris 5123D and 7793 hepatomas. A new paper chromatographic method that separates and quantitates all the major products of the reduction and hydrolysis or other reactions of the substrate has been devised. All of the above tissues were able to reduce UDP and UTP at relatively slow rates ranging from 0.25 nmole of deoxycompound formed (deoxyuridine 5'-triphosphate) per mg protein per hr for liver to 3.5 nmoles deoxyuridine 5'-triphosphate for the Morris 7793 hepatoma when UTP was the substrate. In general, UTP was a better substrate than UDP. The method may also be used to measure cytidine 5'-diphosphate (CDP) reduction, and under the same conditions, the reduction of CDP proceeded at about 6 times the rate of UTP reduction in the Dunning hepatoma. Like CDP reduction, the reduction of UTP was strongly modulated by ATP. Reduction of UTP was insignificant with no ATP or 1.5 micronmoles ATP added to the reaction mixture and was maximal with 0.25 micronmole. The reduction of UTP was inhibited by deoxyuridine 5'-monophosphate, deoxythymidine 5'-triphosphate, deoxycytidine 5'-triphosphate, and deoxyribose 1'-phosphate. The effects of deoxyadenosine 5'-triphosphate varied, depending on its concentration in the reaction medium and whether UDP or UTP was a substrate. However, hydroxyurea did not inhibit reduction of UDP or UTP at concentrations that strongly inhibited CPD reduction. All of the tissues were able to hydrolyze [alpha-32P]deoxyuridine 5'-triphosphate readily to the diphosphate and monophosphate. It is suggested that the enzyme that reduces UTP or UDP may be different in these tissues from the enzyme that reduces CDP.


Growth stimulation of sparse, serum deprived and of confluent, contact inhibited mammalian fibroblasts by a preparation of 3':5'-cyclic AMP phosphodiesterase. Resting mammalian fibroblasts, either sparse and maintained in a serum-free medium, or confluent and contact inhibited, are stimulated to divide by treatment with a preparation of 3':5'-cyclic AMP phosphodiesterase. This enzyme preparation contained a low level of trypsin-like and alpha-chymotrypsin-like activity, but its effect on cell growth could not be mimicked by pure, crystallized trypsin and alpha-chymotrypsin at concentration equivalent to their contamination in the above preparation. Preincubation of the 3':5'-cyclic AMP phosphodiesterase preparation with the protease inhibitor, phenyl methane sulfonyl fulride, did not affect, either, its stimulation of DNA synthesis in fibroblasts, or its enzymatic hydrolysis of cyclic AMP.


Lateral enzyme topology in the rough endoplasmic reticulum of rat liver. Rough microsomes were subfractionated on the basis of different properties in order to investigate the nature and extent of the enzyme heterogeneity of these vesicles. A discontinuous gradient, containing monovalent cations allowed the separation of a ribosome-poor membrane fraction which was enriched in electron transport enzymes and relatively poor in phosphatases. Zonal centrifugation on a stabilizing gradient separated 3 fractions characterized by enrichment of electron transport enzymes, glucose-6-phosphatase and adenosinetriphosphatase, respectively. An essentially similar pattern was seen when ribosomes were removed with EDTA and the denuded vesicles subfractionated on a sucrose gradient. Rough microsomes from phenobarbital-treated rats exhibited the same pattern both qualitatively and quantitatively. It appears that electron transport enzymes and two types of phosphatases are heterogeneously distributed among rough microsomal vesicles.


Morphological equivalent of the bifunctional role of somatostatin. Using the immunoenzyme bridge-technique at the light and electron microscopic levels, somatostatin can be demonstrated in the perikarya of the anterior periventricular nucleus, in the median eminence and in the parvocellular hypothalamic nuclei of the rat. In the latter regions the perikarya are negative, whereas a positive reaction for somatostatin is found in a delicate network of fibers and middle-sized granules of very small axons. In light of these results, the double function of somatostatin-as release inhibiting hormone and as transmitter-is discussed. The positive staining reaction in the organum vasculosum laminae terminalis of male and female rats as well as in the subfornical organ, the nucleus dorsalis thalami and the nucleus medialis habenulae in female controls and pregnant rats is not due to somatostatin-containing structures, but partly to substance P and partlly to a substance which could not be defined.


Disruption of multicellular organization in the cellular slime molds by cyclic AMP. Addition of cyclic AMP causes disorder in the multicellular stage of a number of species of cellular slime molds. In those which produce fruits with cellular stalks, the addition of cyclic AMP stimulates prestalk cells to differentiate into mature stalk cells. Prespore cells do not differentiate into spores under the influence of cyclic AMP, most degenerate and seem to die. I hypothesize that the normal course of differentiation from vegetative cells is one leading to spores, but that cyclic AMP can divert this course to one leading to the stalk cell. Dibutyryl cyclic AMP, cyclic GMP and cyclic AMP disrupt slugs of Polysphondylium pallidum, while species of Dictyostelium are disrupted by only cyclic AMP. The multicellular stage of P. violaceum is unaffected by high concentrations of exogenous cyclic nucleotides. Cell organization of Acytostelium ellipticum, a species with an acellular stalk, was disrupted by cyclic AMP, but no stalk cells were formed; only spores.


The effect of adenosine-3':5'-cyclic monophosphate on the mitotic rate in regenerating Dugesia dorotocephala. Regenerating posterior sections of the flatworm, Dugesia dorotocephala, were treated with varying concentrations of 5'-adenosine monophosphate (AMP), cyclic AMP (cAMP) and dibutyryl cyclic AMP (Bt2cAMP) for 24 h. M/2000 colchicine was added to the medium during the final 4 h of treatment to collect mitotic figures. The mitotic rate was significantly increased at 0.5, 0.1 and 0.01 mM concentrations of Bt2-cAMP. While Bt2cAMP and cAMP produced comparable results at 0.01 mM, only the Bt2-cAMP-treated organisms exhibited a significantly higher mitotic rate at the 0.1 mM concentration. Theophylline and sodium butyrate did not evoke any stimulatory effect on mitotic rate.


Spin label study of detergents in the region of critical micelle concentration. A series of spin probes was employed to examine the behavior of the detergent sodium dodecyl sulfate (SDS) at concentrations above and below the critical micelle concentration (cmc). The existence of detergent aggregates below the cmc was evidenced by the appearance of composite electron spin resonance (ESR) spectra for probes that have measurable solubility in water. The spectra were indicative of two probe populations: one in an aqueous environment and another in detergent aggregates. The ESR spectra of probes which are highly insoluble in water exhibited line broadening due to intermolecular spin exchange interactions, indicating that the probes were concentrated in detergent aggregates present below the experimental cmc. The results are discussed in terms of their significance for the study of the mechanisms of micelle formation and for the detection of detergent aggregates at very low concentrations.


Wilms' tumor: evaluation of histological features. In order to evaluate the factors relating to prognosis, a clinico-pathological study was made in 27 patients with Wilms' tumor. Among these 27 cases, 10 survived more than two years after nephrectomy, 16 died within four days up to two years, and one patient could not be followed up. The younger age group and earlier stage of the disease showed more favorable prognosis. The tumor size, location, and number of tumor nodules had no effect on prognosis. Both the lymphatic and capsular invasions were common in the fatal cases. No vascular invasion was seen in any patient that had preoperative radiotherapy. Among the 10 survivors, six patients had received preoperative irradiation, whose tumors revealed a mesoblastic pattern. Polycystic structures were found in two surviving cases. One survivor's tumor contained prominent squamous epithelium. Another case showed predominance of undifferentiated spindle cells. The undifferentiated mesenchymal type with sarcomatous pattern would not always represent unfavorable prognosis. The tumor with mesoblastic pattern after radiotherapy and those with polycystic structure showed low-grade malignancy and maturation of stromal cells and tubules, respectively.


[Different sensitivities to ultraviolet rays of various functions of the polyoma virus. Stimulation of the synthesis of cellular DNA]. Peritoneal Mouse macrophage were used to study the stimulation of cell DNA synthesis by polyoma virus. Using ultraviolet-irradiated polyoma virus, it was possible to show a difference between the inactivations of infectivity and of induction of DNA synthesis. By statistically analysis of these two phenomena it was found that 39% of the viral genome is necessary for the induction of cell DNA synthesis.


[Age-and sex-related development of ACTH in the pars distalis and neurointermedia of rat pituitary]. The content of ACTH of the pars distalis and neurointermedia of the pituitary of adult Rats is higher in the male than in the female. This differenciation of content, according to sex, appears at puberty at the site of pars distalis; it only appears around the 60th day of life at the site of the pars intermedia. The androgens seem to be responsible for the higher percentages observed in the males.


[Histochemical localization of lactate dehydrogenase in the trout embryo (Salmo irideus, Gibb) during early organogenesis of the cord and the neural tube]. The histochemical study of the LDH in the Trout embryo during the early organogenesis shows a specific localization in notochord cells, in mesodermic cells of the terminal knob and in some prosencephalic neuroblasts. The role of the LDH in the metabolism of NAD as well as in the energetic metabolism of embryonic cells is discussed.


[Immunofluorescence identification of cells with corticotropic and alpha-melanotropic activity in the hypophysis of the grass snake Natrix natrix L]. The use of anti ACTH (17-39) and alpha-MSH allowed us to detect by immunofluorescence in the Pituitary gland of Grass Snake the site of elaboration of ACTH and alpha-MSH. Corticotropic cells are located in the rostral part of the Pars Distalis and in the Pars intermedia. Melanotropic cells occupy the whole part of the Pars intermedia.


[Potentiating effect of cycloheximide on viral interference]. The degradation of the antiviral state can be delayed in vitro by antimetabolites, when added between 5-7 hours after interferon. We explore in this chronological order whether antiviral resistance induced by Newcastle disease virus (N.D.V.) in vivo could be modified by an antimetabolite. Cycloheximide was selected for this study because of its reversible biological effect and lack of toxicity in our experimental conditions. The model system employed was Syrian Hamsters, using N.D.V. as an interferon inducer and encephalomyocarditis virus (E.M.C.) as a challenge virus. A constant and significant increase in survival of animals treated with N.D.V.+cycloheximide is probably related to a delay in the degradation of the antiviral state and not to interferon superinduction.