Effect of vagotomy on the afferent innervation of cat pancreas. The afferent innervation of the pancreas was studied with Holme's silver technique in normal and vagotomized cats 24, 48 and 72 hours after sectioning the vagus. Pacinian corpuscles were found in the interlobular and periductal connective tissue. After right vagotomy, degenerative changes were noticed among some of the nerve fibres entering the formation of the perivascular, periacinar, peri-insular and periductal plexuses. Moreover, degenerative changes were noticed in most of the Pacinian corpuscles while others remained unaffected, thus denoting that the right vagus was not the only pathway of the afferent nerve fibres from the pancreas and probably the left vagus contributed in that domain.


[Evolution of muscles in Nereidae (Annelida polycheta) during epitoky. I and II. Atokous and epitokous longitudinal fibres (author's transl)]. The longitudinal fibres from atokous Nereis belong to the double oblique striation type. Their thick myofilaments, paramyosinic (about 145 A periodicity) are arranged in an hexagonal lattice, well preserved after a glycerol treatment. 9 to 13 thin filaments form an orbit around one thick filament (ratio: 6-7/1). The contraction occurs in two processes: a sliding filament mechanism, and a shearing mechanism. This second mechanism consists in a parallel shifting of thick filaments, arising their overlap and decreasing their degree of stagger; it induces the increase of the myofibrillar (A-bands) oblique angle from 10-12 degrees in isolated glycerol extracted fibres, to 35-38 degrees in the same fibres after contraction by the action of ATP-solution. The longitudinal fibres from epitokous Nereis or Heteronereis are characterized also by a double oblique striation. The structural aspects of the contraction are similar to these ones occurring in atokous fibres. Nevertheless, the epitokous fibres are different from the atokous fibres by many characteristic points. Their thick myofilaments, with an hexagonous array, are thinner and shorter than these ones from atokous fibres. The contractile material is only present in the cortex. The axis of epitokous fibres is filled with numerous mitochondria, between them numerous glycogen particles have been synthetised. The sarcoplasm of the fiber's coelomic edge is often devoided of myofilaments but it is full of mitochondria and glycogen granules. The nucleus, with a voluminous nucleolus, is no more placed in the fibre axis, but in a lateral sarcoplasm containing ribosomes, glycogen and mitochondria. The important transformations of longitudinal fibres are discussed in connection with the Heteronereis locomotor behaviour, very different from this one of the atokus Nereis. Particularly, it seems that the abundance of glycogen and mitochondria in epitokous muscles allows a high contraction frequency. The decrease of the diameter and length from the thick filaments should be set in relation with the augmentation of the contraction's swiftness. These results are very similar to these ones occuring in Syllidae with the stolonization mode of reproduction. The Syllis stolonial fibres present the same characteristics than the Nereis epitokous fibres.


[Relationship between orally induced immune tolerance and local immune response (author's transl)]. BALB/c mice were immunized by intragastric immunization with sheep red blood cells repeated daily for 4 days. This immunization resulted in the appearance of circulating antibodies which were predominantly of the IgA class. When serum from intragastrically immunized mice was administered intraperitoneally to recipient animals 8 h before parenteral immunization with sheep red blood cells, the subsequent immune response was depressed proportionally to the dose of serum injected. When intragastrically immunized mice were challenged intraperitoneally with sheep red blood cells, the level of the IgM response to the parenteral stimulation was in inverse ratio to the IgA response induced by the oral route. These results suggest that orally induced IgA production is related to orally induced immune tolerance and that systemic hyporesponsiveness is achieved while gut plasma cells are producing specific IgA.


Kinetics of haemolysis by complement. ii.--Development of new equations. A new quantitative equation was developed by a process of trial and error. This equation, more complex than the classic Von Krogh equation, nevertheless gives a better account of percentages of haemolysis versus quantities of complement. A basic biological unit independent of red corpuscles quantity and sensitivity can be calculated. In addition, variations of the slope coefficient are of interest in pathology. However, the present paper is devoted to "normal" complement kinetics. Time-kinetics were found to follow the rules of general chemistry and could be derived mathematically.


A light and electron microscopic study of the nucleus isthmo-opticus in the pigeon. A morphological study of the nucleus isthmo-opticus (NIO) has been carried out using several histological approaches as well as electron microscopic analysis. The nucleus is composed of bipolar cells of relatively constant size. The neurones within a given nuclear region are oriented in the same plane and direction. EM observations show that neighbouring cells often give off complex spinous extensions that come in close contact with each other. The apposed membranes of these extensions display large attachment plaques. The neuropil contains relatively abundantly myelinated axons, thin dendrites, and various types of terminals. A majority of the terminals contain rounded vesicles. The tecto-isthmic projection has been studied electronmicroscopically, by using the degeneration technique. The tecto-isthmic terminals contain predominantly rounded vesicles. They almost always make axo-dendritic contacts. Degeneration and autoradiographic techniques at the light microscope level fully confirm previous data by others on the topologic organization of the projection. The above two techniques were applied to the study of NIO-retinal interrelationships. No retinal projection to NIO could be confirmed. On the other hand, enucleation induces a terminal degeneration of slow time course. This finding suggests the presence of intrinsic fibre collaterals of isthmo-retinal axons. The existence of fibre projections from the oculomotor nuclei is shown by a light microscope degeneration study. Fibres coming from the trochlear nucleus might correspond to collaterals of motor axons.


Werdnig-Hoffmann disease. The effects of intrauterine onset on lung growth. Thoracic gas volume (TGV), resting lung volume at end expiration, was measured by the plethysmographic technique in 9 infants with Werdnig-Hoffmann disease. Five of these infants were considered to have intrauterine onset of the disease; the mother in each case had reported a pronounced reduction in fetal activity during the last trimester of pregnancy, and 4 were found to be hypotonic at birth. The remaining 4 infants appeared normal at birth and did not develop any signs of the disease until between 2 and 12 weeks postnatally. Those with intrauterine onset of disease had a significantly reduced TGV (mean 20.8 ml kg(-1)), whereas those with postnatal onset had normal lung volumes (means 36.1 ml kg(-1)). The reduction in lung volume correlated only with intrauterine onset of disease, and was not related to either the degree of muscle weakness or the duration of disease. There is increasing evidence that fetal breathing movements may be one of the essential prerequisites for normal fetal lung development. It is therefore possible that diminished fetal breathing movements, resulting from weakness of the respiratory musculature in utero, could be responsible for the reduction in lung volume found in those infants with intrauterine onset of the disease.


Improved accuracy of lactose tolerance test in children, using expired H2 measurement. Expired hydrogen and blood glucose were measured during an oral lactose tolerance test in 163 children aged between 9 months and 14 years. Lactose malabsorption, defined as an abnormal increase in expired H2 during a lactose tolerance test, was found in 54 children. Of these, 30 were found to be lactose intolerant as the increased expired H2 was accompanied by clinical symptoms. The other 109 children, in whom there was no rise in expired H2, were assumed to have normal lactose absorption. In children with lactose intolerance the increase in expired H2 tended to occur earlier after lactose ingestion than in children with malabsorption. The mean value of the rise in blood glucose was 2.4 mmol/100 ml) in the lactose-tolerant children and 1.0 mmol/1 (18 mg/100 ml) in the lactose-intolerant ones. Although this difference is significant (p less than 0.001), the rise in blood glucose, in predicting the correct diagnosis, was wrong in 13% of cases in the lactose-tolerant group, and wrong in 37% in the lactose-intolerant group (95% confidence limits 9-19% and 22-53% respectively). It is concluded that a rise in blood glucose, whether or not of more that 1.2 mmol/1 (22mg/100 ml) is of little help in differentiating lactose tolerance from intolerance.


Serum theophylline levels in asthmatic children after oral administration of two slow-release theophylline preparations. Serum theophylline levels were measured throughout the day in 65 asthmatic children receiving one of two slow-release theophylline compounds. There was a wide interindividual variation in levels obtained with similar doses of pure theophylline, and also a wide intraindividaual variation in levels throughout the day, suggesting that the rates of release and/or absorption of theophylline from these compounds are not uniform. There were fewer side effects associated with taking these compounds than with choline theophyllinate.


Hypothyroidism in children with cystinosis. Eight children with cystinosis (3 with renal transplants, 2 on maintenance haemodialysis, 2 with chronic renal failure, and one with normal renal function) were studied for evidence of hypothyroidism, and compared with a control group of children with chronic renal failure due to other causes. Abnormal thyroid function was present in all the cystinotic patients: thyroxine (T4) low in 1, free thyroxine index (FTI) low in 2, thyroid-stimulating hormone (TSH) raised in 6; all had a supranormal TSH response to thyrotrophin-releasing hormone (TRH) stimulation, indicating impaired thyroid reserve compared with patients in the control group who had a depressed or normal TSH response. Increased growth velocity with thyroid supplementation occurred in only 2 patients, and the onset of puberty may have contributed to this improvement. Hypothyroidism is a common finding in cystinosis, and it is suggested that thyroxine treatment be started when the TSH concentration becomes raised.


Hypothalamo-pituitary hormone insufficiency associated with cleft lip and palate. Two male patients with congenital cleft lip and palate first seen at ages 10.2 and 21.5 years presented with typical signs of hypothalamic-interior pituitary hormone deficiencies. They were found to lack GH, LH, and FSH and to be partially deficient in TSH and ACTH. Several congenital defects may explain this rare syndrome affecting midline structures in the proximity of the maldeveloped palate, including Rathke's pouch, which migrates distally to develop into the anterior pituitary.


Menkes's syndrome. Report of a patient treated from 21 days of age with parenteral copper. In an infant with Menkes's steely-hair syndrome, early treatment (from 21 days of age) with parenteral copper failed to halt the disease. In addition to urinary tract abnormalities, panlobular emphysema was present a finding not previously noted in the syndrome.


Use of salbutamol powder in childhood asthma. In this study on 31 asthmatic children, salbutamol powder was as effective as salbutamol aerosol in the older asthmatic child studied daily during a 6-week period. The rotahaler can be used successfully from 3 years and, as measured by the peak expiratory flow rate, salbutamol powder is effective in this young age group.


Depigmented hair. The earliest sign of tuberous sclerosis. We have studied 4 children with tuberous sclerosis with one or more patches of depigmented scalp hair and in each case these were noticed by the parents at birth. In one patient the finding of a tuft of white hair preceded the appearance of white macules by many months. A tuft of white scalp hair is a useful new sign of tuberous sclerosis in the newborn and young child, and the hair should be examined as carefully as the skin when early 'organic' seizures are unexplained.


Bone growth in thalassaemic children.

Bone growth in thalassaemic children. X-ray measurements were made of the length and width of the 3 middle metacarpal bones of both hands, in 61 thalassaemic and 35 control children of both sexes aged 5--13 years. Growth in length of the bone was normal until age 11 years but after this it was smaller in thalassaemic children. The growth of total width in thalassaemic children was greater than that of the controls until age 11 years and then became equal. The proportion of children with bone age retardation was greater after 7 years.


Epileptic laughter with precocious puberty. A child with associated epileptic laughter and precocious puberty is reported. The significance of epileptic laughter as a possible marker of hypothalamic disease is noted.


Ecological studies in Madagascar of Biomphalaria pfeifferi, intermediate host of Schistosoma mansoni. 2. Biology and dynamics in the non-endemic area of Antananarivo. The region of Antananarivo, where Schistosomiasis is not yet endemic, is compared to Ambositra, a well established focus of S. mansoni, with regard to climate, hydrology and snail habitats. Higher mean temperatures and a more pronounced dry season in Antananarivo contribute to sometimes less favourable hydrological conditions. Biomphalaria is found in very few and limited habitats. Population dynamics were followed up in selected habitats and showed ecological parameters comparable to those of the Ambositra-strain. Trematode infections were very scarce. Both strains of Biomphalaria are equally susceptible to infections with the malagasy strain of S. mansoni (up to 100 p. 100). Mortality rates, prepatency periods and the electrophoretic pattern of egg proteins are very similar. Foreign snail species like B. glabrata (albino strain) and B. pfeifferi (South African strain) are hardly susceptible to malagasy S. mansoni. Some cases of S. mansoni-infections are mentioned, which presumably had been contracted in non-endemic areas of the province of Antananarivo. The possibility and risk of implantation of schistosomiasis in this region and in the town is evaluated following the discussion of epidemiological details. The author concludes that in spite of some unfavourable conditions for S. mansoni the establishment of the cycle can locally not be excluded. Attention is all the more important, as control of endemic schistosomiasis would be particularly difficult under malagasy conditions of geography, climate, snail dynamics and human behaviour.


[Plesiomonas shigelloides. 23 strains isolated from human stools in Madagascar]. Twenty-three strains of Plesiomonas shigelloïdes isolated from human stools in Antananarivo are studied. Ficklness of some biochemical characters is observed. Ability to pathogenicity is tested on animals (cornea of guinea pigs, intestinal ligatured loops of rabbits).


[Preliminary studies of Conus geographus venom structural properties: evaluation of the toxin's molecular weight]. Venom from the posterior half of the venom duct of Conus Geographus was toxic to mice but without effect on a crab, and produced a flaccid spasmodic paralysis in mice. The minimum lethal dose for mice ranged from 5 micrograms to 50 micrograms (net weight) of venom per mouse. Toxin was submitted to Sephadex G 25 chromatography and to ultrafiltration. The venom was inactivated by pronase. The data suggested that the toxin was a peptide of molecular weight of about 1 500-2 000.


[Alteromonas putrefaciens. 25 strains isolated in Madagascar]. 25 strains of Alteromonas putrefaciens, most of them from marine medium, isolated in Madagascar, are reported. Biochemical characters are studied to class them in 2 subspecies. Routinely, Alteromonas putrefaciens must be chiefly differentiate with H2S-producing Enterobacteriaceae. The ecology is recapitulated.


[Tunisian reptiles. III. Distribution and need for protection of terrestrial and fresh water turtles]. The distribution and habitats in Tunisia of the tortoise Testudo graeca graeca and of the two turtles Emys orbicularis and Mauremys caspica leprosa are investigated throughout the country. The present low level of population density is recent; its causes are precised. The protection, more specially for the two first species, must be very efficient.


[An enzootic with pseudo-tumoral manifestations in Vipera lebetina in captivity]. The authors report the clinical and epizootic evolution of an epizooty, characterized by pseudo-tumours, observed on some vipers (Vipera lebetina) maintained in captivity at the "institut Pasteur de Tunis". The disease was characterized by 2.3 cm size nodules, scattered under the skin or into splanchnic cavities. These ones, spherical and well capsuled, were formed by a mass of clear rounded cells (histiocytes or monocytes), the necrotic central area of which contained innumerable bacteria. Several reviewed works show that this appearance of chronic abcess is a common and univocal reaction to various pathogenic factors, such as bacteria and parasites. As to etiology, the authors have doubt about the pathological role of the germs which they have identified (Salmonella and other anterobacteria) because these latter belong to the normal microbial flora of snakes and are, in the present case, probably occasional infective agents only.


Influence of chlorpromazine on calcium movements and contractile responses of guinea-pig ileum longitudinal smooth muscle to agonists. The effects of chlorpromazine (CPZ) on the uptake and release of calcium by microsomes prepared from longitudinal smooth muscle of guinea-pig ileum (GPI) in vitro and on contractions of longitudinal muscle of GPI in situ to various agonists were studied. CPZ (0.5-1 mM) blocked calcium uptake of the microsomes by 85-95%, and increased the release of preloaded calcium from the mocrosomes from 5% to 40% in the first 5 min. CPZ-induced calcium release was increased by chelating agents [ethyleneglycolbis (beta-amino-ethyl)-N, N-tetraacetic acid and disodium ethylenediaminetetraacetate], KCL and NaCl. These agents increased the CPZ-induced calcium release from 40% in the first 5 min to an average of 87% in the first 5 min. CPZ-induced calcium release was inhibited by CaCl2. CPZ (1-10 micrometer) decreased contractile responses of muscle strips to low and high levels of contractile agonists. These agonists were 5-methylfurmethide (an analog of muscarine), 5-hydroxytryptamine, histamine, BaCl2 and KCl. CPZ moved the dose-response curves to the right and depressed maximal responses. KCl-induced contractions when blocked by CPZ were tested for the shown to be reversed by CaCl2 (9.8 mM). CPZ appears to interfere at 1-10 micrometer levels with calcium movements of the sarcolemma and in vitro at 0.5-1 mM levels with calcium movements of isolated sarcoplasmic reticulum.


The effect of theophylline on the responses of the guinea-pig urinary bladder to nerve stimulation, adenosin 5' triphosphate and histamine. The effects of theophylline (10(-5)M, 10(-4)M and 10(-3)M) on the contractile responses of the guinea-pig urinary bladder to nerve stimulation, adenosin 5' triphosphate (ATP) and histamine have been investigated. Contractile responses were unaffected by 10(-5)M and 10(-4)M therophylline but were significantly reduced by 10(-3)M theophylline. Since responses to histamine were reduced to a similar extent as responses to ATP, it is suggested that theophylline is not a specific antagonist of ATP in this tissue and that the ATP receptors in the bladder differ from those in other smooth muscle preparations.


[Attempts at inter-specific hybridization in Sphaeroma (Crustacea Isopoda Flabellifera)]. An interspecific hybridization between five tunisian species of the genus Sphaeroma has been attempted. All the trials failed whatever the crossing attempted. The lack of natural hybrids between forms living together constitutes a fundamental criterium for specific discrimination.


[Demonstration of an enzyme inhibiting coagulation of blood ingested by the blood-sucking insect, Rhodnius prolixus (Heteroptera, Reduviidae)]. A new fibrinolytic enzyme, "the prolixase", was isolated and purified from the haemolymph of Rhodnius Prolixus. The mode of action of the enzyme was studied and compared to that of extracts from salivary glands and midgut of the insect. It appears that the prolixase is involved in inhibiting the coagulation of the blood ingested by Rhodnius during its blood meal. The mode of action of the enzyme in this phenomenon is discussed.


The inhibitory effect induced by delta9-tetrahydrocannabinol on the contractions of the isolated rat vas deferens. To analyse the influence of delta9-Tetrahydrocannabinol (THC) on the effects induced by sympathomimetic, parasympathomimetic and direct spasmogenic drugs, the isolated rat vas deferens preparation was chosen. Instead of the usual emulsifying agents, we have used 1% ethanol (ET-OH) to keep the cannabinoids in homogeneous dispersion in the bath. THC exerted a non-competitive inhibition of noradrenaline and of acetylcholine responses but no synergistic action nor competitive antagonism was seen on the dose range used. BaCl2 was potentiated by ethanol and this effect was abolished by THC. The water soluble derivative SP 111 showed actions similar to those of THC on noradrenaline and acetylcholine responses. It did not affect BaCl2 contractions, indicating that the cannabinoid might act by depressing early events of membrane activation.


Action of salicylate on failing heart lung preparations of guinea-pig perfused with red blood cells suspended in Tyrode-dextrane. Heart lung preparations (HLP) of guinea-pig were perfused with homologous red blood cells suspended in 1.5 volumes of Tyrode containing 6% dextrane. Red blood cells were freshly obtained by centrifugation of heparinized blood and subsequent washing with saline. Reduced pulmonary expansion due to bronchoconstriction, increased pulmonary vascular resistance and heart failure rapidly occurred in these preparations. Addition of salicylate to the perfusing medium delayed and attenuated both the respiratory and the cardiac disturbances. A full protection against cardiopulmonary failure was obtained when salicylate was added to the heparinized blood used for red blood cells preparation and to the saline used for red blood cells washing. It is concluded that pulmonary microembolism, due to platelet aggregation, is the primary factor responsible for HLP failure. Bronchoconstriction and pulmonary vasoconstriction are aggravating factors secondary to microembolism. Salicylate appears to prevent the microembolism and the microembolism depending release of broncho- and vaso-constrictor autacoids.


[Serodiagnosis of toxoplasmosis in man]. We want to repeat the interest of toxoplasmosis serodiagnostic in human pathology. First, to help the diagnostic of this affection, with adult and children; then, to prevent this parasitose with pregnant women. Results in our laboratory, since two years, point out that a large number of women are seronegative. We are proposing a protocol of supervision for women before and during their pregnancy. This protocol avoids the risk of foetal contamination and prevents congenital toxoplasmosis.


[Host-parasite interactions between Coccus hesperidum L. (Hom. Coccidae) and its parasite Coccophagus scutellaris Dalman (Hym. Aphelinidae)]. The eggs laid in the host gut will develop normally. The larva resulting from those eggs penetrate the hemocele through the gut wall. The structure of the cells near the entry point is not affected because of the continuous renewal of those cells. On the other hand, development of the ovaries is most seriously affected. In fact, ovogenesis is stopped and vitellogenesis slowed down. But this phenomenon is related to the initial level of ovary development. If the attack takes place when the host is young, the effect on the ovaries is more serious. In extreme cases gonad development is annihilated. Otherwise, the host will still be able to lay eggs, but its reproduction capacity is diminished. Factors reducing development of gonads or maintaining them at a juvenile stage are discussed.


Absorption and disposition of N-(1, 1-dimethylethyl)-N'-[2-(4-pyridinyl)-4-pyrimidinyl]urea in rats and dogs. N-(1, 1-Dimethylethyl)-N'-[2-(4-pyridinyl)-4-pyrimidinyl]urea, Win 40, 882, was eliminated from the blood stream of dogs by two apparent first-order processes with alpha- and beta-phase half-lives of 0.2 hr and 1.4 hr, respectively. Radioactivity of the administered dose was excreted by rats in the feces and via the kidneys; about 40-45% of the dose was recovered in the feces, with the remainder in the urine, over a six day period. One of the terminal methyl groups of the tert-butyl moiety of Win 40,882 is sequentially oxidized by the rat to the alcohol, aldehyde and carboxylic acid. In addition, some of the aldehyde was further metabolized to generate two different monohydroxylated aldehydic metabolites; these hydroxyaldehydes accounted for less than 10% of the dose administered. An unusual metabolic pattern was noted in the excretion of Win 40,882. Over 30% of the urinary metabolites contained the carboxaldehyde function; only8% of the urinary radioactivity was represented by the further oxidation of the aldehyde group to generate the carboxylic acid.


[In vivo titration of tetanus antitoxins]. The theoretical background and the practical techniques of assay of tetanus antibodies on mice by the L+ toxine procedure, is precisely described in detail. Basing the arguments on certain aspects of the method, the accuracy of the test is estimated and its reproducibility is assessed, in terms of both in vivo and in vitro titrations.


Interaction of loperamide and diphenoxylate with ethanol and methohexital. The antidiarrheal drugs loperamide and diphenoxylate were tested for their ability to potentiate central nervous system depression induced by ethanol in mice and nethohexital in rats. Oral diphenoxylate potentiated the loss of righting reflex (hypnosis) at 10 mg/kg in mice and at 5 mg/kg in rats, doses which were considerably lower than those required to induce morphine-like behavior in mice and rats. Oral loperamide did not potentiate hypnosis. Increased incidence and duration of hypnosis was observed with loperamide only at very high, nearly toxic doses (100 mg/kg in mice and 80 mg/kg in rats) and these doses induced only nonspecific central nervous system depression. The failure of loperamide to potentiate the action of hypnotics confirms data from clinical trials which showed that loperamide is essentially free of central depressant effects when given orally.


Effect of ethanol, pentobarbital and morphine on the toxicity of hemicholinium-3 and d-tubocurarine in mice. Pretreatment of mice with either ethanol, pentobarbital or morphine protected against the lethal effects of hemicholinium-3 (HC-3) but not curare. The protective action of these compounds was not due to an induced release of corticosteroids in vivo since administration of prednisolone did not protect against HC-3 toxicity. The protective action of morphine was reversed by Nalline hydrochloride. The results suggest that the site of the protective action of ethanol, pentobarbital and morphine and the site of action of the lethal respiratory depression by HC-3 are central in origin.


Pharmacological and histological studies of gastric mucosal lesion induced by serotonin in rats. Serotonin in doses of 5 to 80 mg/kg injected subcutaneously to rats injured the gastric mucosa dose-dependently and the lesion was histopathologically regarded as erosion. Aggregated platelets were observed in small blood vessels of the gastric mucosa prior to the development of erosion when serotonin was injected in a subcutaneous dose of 20 mg/kg effective in producing the lesion. Subsequently fibrinous thrombi could be found in the lesional mucosal area at the stage of erosive change. Serotonin in the same dose decreased gastric mucosal blood flow and enhanced gastric motility in the peroid of time when pre-erosive or erosive change occurred. Circulatory disturbance of the gastric mucosa induced by these histological and functional changes seemed to play an important role in development of the gastric mucosal lesion by serotonin. It is unlikely that gastric acid takes part as a trigger in producing the lesion.


Regional differences in the effects of denervation, cocaine and chronic reserpine administration on the responses of the rat vas deferens to norepinephrine and acetylcholine. The prostatic half and epididymal half of the rat vas deferens was found to show the difference with respect to both the sensitivity (expressed as the geometric mean ED50) and maximal response to norepinephrine and acetylcholine. The epididymal half was more sensitive to both drugs than the prostatic half; the maximal response to norepinephrine was greater in the epididymal half than in the prostatic half. Effects of denervation, 10-6 M cocaine and chronic administration of reserpine (1 mg/kg/day for 5 days) on the responses to the drugs were also found to be qualitatively and/or quantitatively different between the two halves. In the epididymal half, all these procedures produced supersensitivity to acetylcholine as well as to norepinephrine. Whereas, in the prostatic half, supersensitivity to only norepinephrine was observed; the responses to acetylcholine were more or less depressed, accompanied by little change in the sensitivity, by these procedures. Degree of supersensitivity (ED50 control/ED50 treated) to norepinephrine produced by denervation was greater in the prostatic half than in the epididymal half. However, in the case of cocaine or reserpine, there was little difference in the degrees of the two halves. In both halves, the maximal response to norepinephrine was increased by denervation and cocaine, but not by reserpine. The magnitude of such increase in the prostatic half was greater and less, after denervation and cocaine, than that in the epididymal half, respectively. Only cocaine increased the maximal response of the epididymal half to acetylcholine. These results are discussed in relation to those previously obtained using whole vas deferens.


Motility and hemodynamics of the canine gastrointestinal tract. Stimulation by pentagastrin, cholecystokinin and vasopressin. Effects of pentagastrin (1-4096 ng/kg), cholecystokinin (1-4096 mU/kg, CCK) and vasopressin (.032-128 mU/kg) on gastrointestinal motility and blood flow, were determined by simultaneous measurement of blood flow (electromagnetic flow probes) to and motor activity (strain gages) of corpus, antrum, duodenum, jejunum, and colon of anesthetized dogs. Antral contractile amplitude was increased by pentagastrin at relatively low doses. Pentagastrin also increased corpus blood flow, corpus tone and antral blood flow. Gastric contractile frequency was least sensitive to pentagastrin. Corpus blood flow was decreased and small intestinal blood flow was increased by cholecystokinin at relatively low doses. CCK also increased small intestinal contractile amplitude and, at higher doses, antral contractile amplitude, and duodenal tone. Time-effect relation and sensitivity were different for the hemodynamic and motor responses to pentagastrin and to cholecystokinin. This shows the lack of correlation between vasoactive and motor-stimulating properties of these drugs. However, strong drug-induced contractions were shown to impede antral blood flow (pentagastrin and CCK) by about 35% and duodenal and jejunal blood flow (CCK) by resp. 70 and 60%. Vasopressin reduced blood flow to stomach and intestines by 50-80%, without affecting gastrointestinal motility.


Influence of frequency of stimulation on the toxicity of digoxin on isolated guinea-pig atria in different extracellular Ca2+. In isolated guinea-pig left atria driven by rates of 0.5, 1,2, 3 and 4 Hz digoxin was infused continuously until cardiac arrest occurred. The experiments were performed in Tyrode solution with normal and twice the calcium content. Calculated was the toxic concentration of digoxin as that concentration which caused arrest of the preparation. In normal Ca2+-content (1.8 mM) an increase of the driving frequency decreased significantly the toxic concentration of digoxin, i.e. elevated the toxicity of this cardiac glycoside. The elevation of the Ca2+-content of the Tyrode caused a further diminution of the toxic concentrations of digoxin in dependence of the driving rate, thus demonstrating a synergism of Ca2+ and digoxin with regard to its toxicity.


Drug action on cerebral energy state during and after various hypoxic conditions. The behaviour of fuels (glycogen, glucose), of glycolytic pathway intermediates (glucose-6-phosphate, pyruvate) and end-product (lactate), as well as the pool of labile phosphates (ATP, ADP, AMP, creatine phosphate) and the energy charge of the brain were studied in the motor area of the cerebral cortex of beagle dogs. These parameters were evaluated both after various hypoxic conditions (hypoxic hypoxia, hypoxia plus complete or incomplete ischemia) and after 3, 15 or 30 min of post-hypoxic recovery and recirculation. The effect of some drugs (papaverine, UDP-glucose, (-)eburnamonine, suloctidil) following intracarotid perfusion has been evaluated in the various quoted experimental conditions. The tested drugs proved unable to improve the deranged brain metabolism under all the hypoxic conditions. On the contrary, an activating effect of suloctidil and (-)eburnamonine could be observed during the recovery after both hypoxia and hypoxia plus complete ischemia, papaverine being ineffective and UDP-glucose increasing the glycogen synthesis. The drugs proved unable to induce a restitution of the altered brain metabolism after hypoxia plus incomplete ischemia.


The effect of aggregation on the lethality of morphine in mice. Morphine was more lethal to mice housed in groups (aggregated) than to mice housed individually (isolated). The aggregation effect on morphine lethality was increased when ambient temperature was increased to 29 degrees C and abolished when ambient temperature was lowered to 19 degrees C. At 29 degrees C morphine produced hypothermia in isolated mice and hyperthermia in aggregated mice. Both isolated and aggregated mice were hypothermic after morphine administration at 19 degrees C. At 29 degrees C the locomotor activity of morphine-treated aggregated mice was significantly greater than that of morphine-treated isolated mice, whereas at 19 degrees C no significant difference was found. The locomotor activity time course of morphine-treated aggregated mice at 29 degrees C closely resembled the rectal temperature time course, which suggests that the hyperthermia in those animals involves an increase in heat production via an increase in locomotor activity. However, the hyperthermia observed was not sufficient to explain the increased lethality in aggregated mice. Differences between the aggregation effect on morphine lethality and that previously reported for amphetamine were discussed.


[A new type of sialidosis with kidney disease: nephrosialidosis. I. Clinical, radiological and nosological study]. The term nephrosialidosis is proposed to describe a type of oligosaccharidosis in which a glomerular nephropathy develops early and causes death in the early years of life. The clinical and radiological features of the disease are dysmorphic facies, visceral storage disease, early and severe mental retardation and skeletal abnormalities of a type often described in this group of diseases. Foam cells were present in the marrow and, late in the illness, a cherry red spot was present on fundoscopy. The condition is inherited as an autosomal recessive. The leucocytes were deficient in alpha-(2-6) neuraminidase, an abnormality that has also been described in mucolipidosis type I and in other conditions quite distinct from nephrosialidosis. Thus the conditions characterised by this enzyme deficiency are definitely heterogeneous.


[A new type of sialidosis with kidney disease: nephrosialidosis. II. Anatomic study]. The anatomical, macroscopical, histological, histochemical and ultrastructural findings in nephrosialidosis have certain similarities to those found in mucolipidoses. (Excess of complex lipid in neural tissue and an excess of light coloured material in the reticulo-endothelial system that is not easily characterised). Several distinct features help to distinguish nephrosialidosis from closely related conditions (renal lesions, storage of material in sympathetic ganglia).


[Congenital "cardio-esophageal" syndromes. Congenital cardiopathies and multiple abnormalities syndromes in the child with esophageal abnormalities]. A case of esophageal stenosis and tracheo-esophageal fistula associated with dextrocardia due to dextrorotation, ventricular septal defect and complete double arotic arch is reported. In 116 cases of esophageal atresia and tracheo-esophageal fistula 9 had an associated cardiac malformation (4 right-sided aortic arch and 4 ventricular septal defects). The possible individualization of multiple abnormality syndromes in which these two types of malformations are associated with others such as vertebral arch defects, limb anomalies, imperforate anus, etc. is discussed. These are so-called "Vater" or "Vactel" syndromes.


[Enteropathic acrodermatitis treated with zinc sulfate]. A six month old baby with acrodermatitis enteropathica was treated successfully up to the age of 2 1/2 years with oral zinc sulphate, 100 mg daily. No complications were seen but there were two relapses. The first was at the age of 15 months when the dose of zinc sulphate was reduced to 50 mg daily and the second at 20 months when the zinc was stopped by mistake.


[Thyroid involvement in cystinosis].

[Thyroid involvement in cystinosis]. Thyroid function was assessed in 15 children with cystinosis and it was abnormal in 12. Three of these children were clearly hypothyroid and in the other 9 there were abnormalities of TSH or in the response to TRF. Compensated hypothyroidism is frequent in cystinosis and may be one factor in the severe growth retardation in this condition. The preliminary results of replacement therapy support this hypothesis.


[Weight, height, skull and chest circumferences, biparietal diameter in the newborn infant as a function of gestational age, weight, height and weight gain of the mother]. The weights, heights, skull and chest circumferences and biparietal diameters were measured in 7,905 newborns in 1972, in the 5 maternity hospitals of Lyon. These data were studied with respect to the gestational ages of the children and according to the usual weights, heights and weight gains of the mothers. The evolution of these parameters was compared to each other; tentative explanations are given.


[Late lymphedema in Turner's syndrome].

[Late lymphedema in Turner's syndrome]. Two cases of delayed unilateral lymphedema are reported in children presenting with Turner's syndrome. On this occasion, the frequency of lymphedema in Turner's syndrome and the more general problem of the lymphatic changes in this syndrome are discussed. The importance of isotopic lymphography is emphasized, because of its simplicity, its harmlessness and the good quality of the morphological and cinetic data obtained.


[Arhinencephaly detected by a pitressin-sensitive diabetes insipidus]. A 4-month old child presented with facial malformations and severe hypernatremia. Hypernatremia was secondary to diabetes insipidus due to a disorder of ADH secretion, associated with cerebral malformations. Clofibrate treatment was ineffective. However, after the patient was treated by a low osmotic residue diet, an increased water-intake and hydrochlorothiazide, natremia became normal and growth resumed.


[Proliferation of synovial fluid cells (author's transl)]. Investigations with labelled thymidine indicated that the mononuclear cells of the synovial fluid only rarely proliferateunder physiological conditions. Even in cases of rheumatoid arthritis no elevated proliferation rate was found. Cytophotometric investigations on Feulgen-stained nuclei is another way to determine cellular proliferation. To answer the question if the relative content of neutrophilic granulocytes in synovial fluids influences the proliferation of mononuclear cells, the DNA content of these cells was determined after Feulgen-staining with respect to the number of neutrophilic granulocytes. It was observed that nearly all mononuclear round cells exhibit diploid nuclear DNA content. No effect of the intensity of inflammation (as expressed by the content of neutrophilic granulocytes) was observed. From these results it is concluded that mononuclear cell proliferation is not influenced by the intensity of the inflammation. On the other hand it can be assumed that there is no immunological stimulus for an increassed proliferation of the synovial fluid in rheumatoid arthritis.


[Clinical assessment of the activity and results of therapy in joint diseases through cytologic examination of puncture fluids (author's transl)]. Activity and results of therapy of joint diseases can be evaluated by means of cytological examinations of the synovia which is at the same time a prerequisite for differential diagnosis using the puncture fluid. Our cytological examinations of more than 700 puncture fluids stemming from different joint diseases were carried out immediately after the puncture on the fresh material, in order to prevent misinterpretation. This may occur through flaking out in stored material, which results in decrease in the cell count and a non-representative differential cell picture in the supernatant, as well as through degeneration of cells. In two thirds of our patients the local process activity, measured in terms of the cytological phase of activity, became evident in the total clinical activity. The synovia analysis furthermore permits objective assessment of the result of intra-articular injection treatment. Intractable activity, substantiated through synovia examination indicates synovectomy.


[Cell reactions in the cerebrospinal fluid with inflammatory and neuroallergie diseases of the central nervous system (author's transl)]. In the course of bacterial and abacterial meningoencephalitis the cell reaction of the cerebrospinal fluid can well be surveyed, permitting important conclusions to the reactions occurring in the leptomeningeal vascular connective tissue. The application of additional cytological methods in completion of the panoptic routine staining proved to be favourable. The respective results may be additionnally availed of for the diagnostic confirmation of the activities found in the panoptic routine cell-picture. In addition to the general evaluation of the nitro-blue tetrazolium test, pyroninophilia, peroxidase and PAS-reaction, above all, the observation somehow the despositing of the reaction products in the celles has proved to be important for the activity detection of the nonspecific esterase. For judging the immunreactive processes in the central nervous system, the role of the lymphocytes and their forms of transformation, as well as their behaviour with the different cytological reactions has to be duly considered.


[Chemotherapy in Hodgkin's disease (author's transl)]. The chemotherapy in its systemic form should be reserved for advanced Hodgkin's disease, i.e. stages III B and IV, but it deserves consideration even for stage II B if B-symptoms are prominent. The following problems will be discussed: 1. Monotherapy or combination chemotherapy? 2. Simultaneous or sequential drug application in case of combined chemotherapy? 3. Continuous application or intermittent therapy? 4. Maintenance therapy or unmaintained remission? 5. Use of chemotherapy in early stages too? 6. Use of combined modality approach (chemo-/radiotherapy) with regard to the stage of the disease? Some combination chemotherapy regimens (MOPP, ABDV) as well as sequential drug application in case of treatment failures will be discussed . Hazards of a complete course of chemotherapy following total nodal irradiation or total nodal irradiation following such combination chemotherapy are mentioned. If the radiation dose is reduced, the combined modality approach might still improve results of treatment. Finally some new drugs in development for treatment of resistant disease are introduced.


[Laparotomy and splenectomy in Hodgkin's disease (author's transl)]. Results of staging laparotomy in 62 patients between 12 and 71 years old are reported. Only in one case postoperative complications were seen. In 27 percent of case the clinical stage was corrected by surgery, mostly from II to III.


[Radiation treatment of Hodgkin's disease (author's transl)]. Radiation treatment of Hodgkin's disease--especially in stages I and II when it may heal the disease--requires thorough radiation planning. It is recommended to use telecobalt or ultrahard x-rays of a betatron. For superficial lymph node regions, electrons of the betatron are useful.


[New cytostatics and their clinical application (author's transl)]. A comprehensive review of the state of development and clinical evaluation of cytostatic agents is given. Special attention is directed to agents which are known for long a time but whose clinical importance has grown only in recent years. This review takes into consideration the chemical structure, mode of action, pharmacokinetics, dosage, side effects, and results of clinical treatment of the following drugs: adriamycin, nitrosourea preparations, DTIC, dibromdulcitol. trofosfamid and ifosfamid, ftorafur, 5-azacytidin, platinum complexes, ICRF-159, cytembena, HMM, VM 26 and VP 16-213, cytostasan, estramustinphosphat, antibody-cytostatic complexes.


Effect of maternal nutrition and socio-economic factors on birth weight of babies in Bangladesh. One hundred post-partum women and their newly born infants were studied in a local Maternity and Child Health Center. A consistent relation appears to exist betwen mother's weight and birth weight of infants. A significant relationship also exists between birth weight and socio-economic status of mothers as well as her parity. It is concluded that improvement in nutritional status during pregnancy would lead to a substantial decrease in the incidence of low bith weight infants in this country.


Birth weight, length, head and chest circumference and haemoglobin level of new born. The birth weight, length, head and chest circumference and haemoglobin level of 1002 full term new born babies (506 males and 496 females) were determined. The results were compared with similar data from other countries. The birth weight of the present series of newborns were similar to those of newborns from India, the crown-heel length agreed favourably with the data from India and Indonesia, and the head circumference were almost equal to those of the newborns from Sri Lanka, Jamaica and India.


Change of serum protein level during pregnancy and the impact of parity and diet on it. Serum protein level in the three trimesters of pregnancy and the impact of parity and dietary protein on it were studied in 154 pregnant women. The mean total serum protein during first, second and third trimesters were found to be 6.85, 6.60 and 6.81 gm% respectively whick were lower than the mean value obtained in a group of nonpregnant women (7.55 gm%). The level of serum protein depends on intake of dietary protein and influenced by parity in pregnant women. While primigravidae with adequate protein intake had higher serum protein level than primigravidae with inadequate protein consumption, multigravidae with adequate protein intake had lower serum protein level than primigravidae with adequate protein intake.


Dibucaine number (DN) in duodenal ulcer. Dibucaine Number (DN) was determined in 103 patients suffering from duodenal ulcer diagnosed radiologically and 214 normal subjects (control). Statistically significant (p less than 0.001) difference was observed in the distribution of DN in the two groups. Whereas 94% of the control subjects had normal DN (above 70), 6% had intermediate DN (between 40--70) and none had atypical DN, only 32% of patients suffering from duodenal ulcer had normal DN, 64% had intermediate DN and 4% had atypical DN. The results have been discussed.


Effect of anabolic steroid on the development and treatment of rickets in rats. The influence of anabolic steroid on the process of production of rickets in vitamin-D deficient animals as well as its influence on healing of rickets with vitamin-D therapy was studied. Long-Evans strains of female rats of two age groups (6 weeks and 4 weeks) were given rachitogenic diet and were divided into five sub-groups-(I) control receiving vitamin-D from the 1st day, (II) rachitic group, (III) receiving vitamin-D from 36th day, (IV) receiving anabolic hormone from the beginning of the experiment and (V) receiving both vitamin-D and anabolic hormone from 36th day of treatment. It was observed grossly as well as histologically and radiographically that: (1) the anabolic steroid minimizes the rachitic changes in the bones of vitamin-D deficient rats, (2) in rachitic animals, a combination therapy of anabolic hormone and vitamin-D results in comparatively greater calcification of osteoid matrix and better healing and remodelling of bone than with vitamin-D alone.


Psychiatric morbidity in general practice. In order to estimate the psychiatric morbidity in patients asking for consultation with general practitioners, a study was carried out in a general practice in Dacca. This revealed a considerable psychiatric morbidity. The two most frequently encountered psychiatric conditions were neurotic and psychosomatic disorders.


Analysis of the epidemiological data of 600 psychiatric patients. Analysis of a series of 600 psychiatric patients randomly selected from those attending a psychiatric clinic reveals that every single group of people shows its quota of psychiatric illness and all the major forms of illness are represented in a mixed society. From the present analysis it appears that Hindu patients are more careful for treatment than other religious groups and female patients were less frequently brought for treatment than their male counterpart. Patients coming from urban areas have many advantages over their rural counterpart in seeking scientific psychiatric treatment. Schizophrenia, affective disorders and anxiety neurosis were the three most common illnesses and these contributed to 77% of the total number of patients.


Changes in the process of ageing--Part--II. Sodum, potassium, calcium and ascorbic acid level in blood of 62 elderly persons (34 males and 28 females) aged between 45 to 80 years of age were determined and compared with the blood levels of 40 (20 males and 20 females) young persons aged between 25 to 38 years of age. No significant difference was observed in the blood levels of elderly males and females or between elderly persons and young persons.


A mutational approach to the study of photorespiration. A number of quite different types of variant cell lines of tobacco have been isolated. This multiplicity of approaches has been in part dictated by the uncertainities of the photorespiratory pathway. We are in the process of determining the relationship between the observed alterations and the glycolate pathway. The INH-resistant trait has been observed in callus derived from plants regenerated from the original resistant callus and in cone case from seedlings of the subsequent generation. It is hoped that these cultures and the plants derived from them will further our understanding of photorespiration and will eventually lead to the ability to regulate photorespiration and net photosynthesis in higher plants.


The opportunity for and significance of alteration of ribulose 1,5-bisphosphate carboxylase activities in crop production. Increased population and the dietary changes accompanying increased affluence are creating a need for a suggested doubling of world cereal grain production (a 3% per year compounding rate) and quadrupling of grain legume production (a 6% per year compounding rate) during this quarter century (1). CO2 enrichment of field-grown crops has demonstrated the possibility of enhancing RuBP carboxylase activity to achieve improved crop production; it increases the production of grain legumes by 50 to 100% and that of cereal grains, for which the studies are less complete, by perhaps 10 to 50%. Results of O2 alteration of growth-room legumes and cereal grains are consistent with the results of CO2 enrichment except for a second role of O2 in assimilate partitioning. It may be necessary to include other components of the system, e.g., additional soil fertility, especially for non-N2-fixing plants, to enable an improved RuBP carboxylase to increase production. No practical method--chemical, genetic, or physical--of improving RuBP carboxylase activity has been reported.


Mechanism of action of ribulose bisphosphate carboxylase/oxygenase. RuBP carboxylase-oxygenase appears to catalyze carboxylation and oxygenation by homologous mechanisms. A common binding site exists on the enzyme for the acceptor substrate, RuBP. A mechanism is proposed whereby RuBP is isomerized, and a carbanion is generated at C2. Then, either CO2 or O2 is added as an electrophile at C2 to form the corresponding 3-keto-2-carboxy-RBP or 3-keto-2-hydroperoxy-RBP adduct. Hydrolytic cleavage at the C2-C3 bonds of these intermediates by the enzyme is envisioned to produce 2 molecules of 3-phosphoglycerate in the carboxylation sequence and 1 molecule of phosphoglycolate and 1 molecule of 3-phosphoglycerate in the oxygenation sequence. Further work will be necessary to establish the validity of the proposed mechanism.


Regulation of photosynthetic carbon assimilation. It may be concluded that the conversion of PGA to DPGA plays a key role in induction and in the regulation of cycle activity. The high concentrations of PGA in actively photosynthesizing chloroplasts reflect this role and the control exerted by adenylate ratios. Thus the cycle can operate at its maximum rate only in the presence of high PGA and low ribulose 5-phosphate concentrations. Once induction is complete, the reductive pentose phosphate pathway will continue to function at its maximum rate if sink activity within the cytoplasm makes available sufficient Pi to support rapid export of triose phosphate. If triose phosphate tends to build up in the straoma, it will favor pentose monophosphate accumulation. A relative excess of ribulose 5-phosphate would, in turn, inhibit PGA reduction (and hence its own formation) by drawing too heavily on the available ATP.


Effects of enhanced coronary flow during cardiopulmonary bypass: development of myocardial edema. Coronary blood flow, left ventricular diastolic stiffness, ventricular water content and ventricular thiocyanate space were investigated in empty fibrillating and empty beating dog hearts during cardiopulmonary bypass. In empty fibrillating hearts coronary blood flow was considerably enhanced even at levels of normal perfusion pressure. A close correlation was demonstrated between enhanced coronary flow, left ventricular diastolic stiffness, tissue water content and thiocyanate space.


Effect of propranolol on enzymatic and histochemical estimates of infarct size in experimental myocardial infarction. Acute myocardial infarction was produced in anesthetized dogs by ligation of the left anterior descendens coronary artery. Propranolol (2 mg/kg i.v.) administered 4 hours post ligation was examined for its ability to reduce infarct size estimated by histochemical and enzymatic methods. There was a signficant correlation between these two methods in their estimation of infarct size. Treatment with propranolol significantly decreased infarct size estimated with both methods. It is concluded that some portion of the myocardium can be protected against infarction by pharmacologic intervention as late as 4 hours after the onset of coronary artery occlusion.


Exerimental coronary artery occlusion. III. The determinants of collateral blood flow in acute coronary occlusion. Coronary collateral blood flow was measured with tracer microspheres in 3 different experimental conditons in the dog heart: 1. after occlusion of a large coronary artery in the in situ beating heart, 2. after occlusion of a small coronary artery in the in situ beating heart and 3. after occlusion of a large coronary artery in the isolated, empty beating, blood-perfused heart. with large coronary artery occlusion in situ, collateral blood flow was very low (5ml/min/100 g). Large coronary artery occlusion in the isolated heart showed a much higher collateral blood flow (25 ml/min/100 g). Small coronary artery occlusion in situ showed a relatively high collateral blood flow (22 ml/min/100 g). Small coronary artery occlusion produced only very small infarcts even when infarct size was expressed as the ratio of infarcted area over the area-at-risk (i.e., perfusion area of the occluded artery). Our experiments showed that acutely available collateral blood flow depends on perfusion pressure, extravascular resistance, size of the area at risk and size, number, and location of collateral vessels.


Mast cell leukemia and acute basophilic leukemia. Cytochemical studies. Human mast cells and basophil granulocytes can be easily recognized in normal tissues by light microscopy. In one mast cell and one basophilic leukemic case considered in this study, mast cells and basophils were morphologically quite similar and could not therefore be clearly defined merely by their morphological features. Both types of cells showed round nuclei and deep purple granules. The diagnosis of mast cell leukemia or basophilic leukemia was made on the basis of different cytochemical patterns. In the case of mast cell leukemia, peroxidase and PAS stains were negative, while chloroesterase was strongly positive; in the case of basophilic leukemia, peroxidase and PAS stains were positive, while chloroesterase reaction showed a peculiar pattern. Toluidine blue metachromasia and astra blue positivity were present in the cells of both cases.


Kinetically oriented manipulation of drug management in acute leukaemia. The evaluation of the kinetic characteristics of the blast population in AL, at the outset of the disease, establishes its growth rhythm and hence the degree of invasivity. Cases with a higher 3H-Tdr labelling index also display a higher growth fraction, birth and growth rates and a shorter generation time, in other words, greater cell growth potential and invasivity. The labelling index could thus represent an immediate guide to prognosis. Particularly when combined with other prognosis factors, it can be employed to divide AL into high and low growth and invasivity forms. A distinction of this kind can thus be made the basis of differentiated therapeutic programming, whereby more or less aggressive cytocidal drug associations and doses can be administered to improve the therapeutic index and the quality of life.


Impedance cardiography the reproducibility of stroke volume measurements under conditions of mass examination. The stroke volume at rest of 81 test subjects with no evident cardiac or circulatory dysfunction was determined by impedance cardiography and the measurements were repeated after 11 weeks. In order to attain comparable and reproducible conditions, 44 test subjects were chosen who showed heart rates at rest not higher than 83 beats per min, and whose changes in heart rate between the two tests were not greater than 11 beats per min. The stroke volumes determined by impedance cardiography are greater than those measured with the invasive methods reported by other authors. If the electrodes are removed between the tests sufficient reproducibility is not obtained. It is concluded that determinations of stroke volume by impedance cardiography as described in the literature are only suitable for detecting directional changes when the electrodes are left in place. It is impossible to obtain comparable data with electrodes once removed and replaced for the second measurement. Impedance cardiography is based on theoretical assumptions which do not permit estimation of errors affecting the measurement.


Lymphomas of the spleen and bone marrow lymphoid nodules. Many nodular primitive non-Hodgkin's lymphomas of the spleen have a favourable course after splenectomy or chemotherapy. Several observations of this type have been reported in the literature, in which the concept of malignancy is discussed, authors referring to a presarcomatous state or to an idiopathic splenomegaly. The evaluation of the extension of those sarcomas, however, very often show hepatic lesions, and always an increase in the lymphoid marrow nodules. The significance of these nodules is discussed here, with reference to 14 personal observations. These nodules may not always reflect a real extension of the sarcoma to the marrow. From a practical point of view, the presence of lymphoid marrow nodules, when associated with an isolated splenomegaly, is a strong argument to suspect a sarcoma of the spleen, and indicates a splenectomy.


Histological prognosis in aplastic anaemia. In the prognosis of aplastic anaemia, bone marrow biopsy displays several data which were unknown by the myelogram: quantitative evaluation of marrow cellularity; vascular and mesenchymatous lesions; extension of lymphoid or histiocytic infiltration. Nevertheless, the confrontation and value of these signs with the previously used prognosis data issued from clinical and haematological observations is still discussed. The lack of a clear understanding of the bone marrow failure delimitation is perhaps the reason of these differences. We present the histological results of a cooperative protocol concerning 350 cases of bone marrow failure with 220 first bone marrow biopsies examined in triple blind and confronted in a statistical comparison with the mortality in the first 20 months. We evidence the early unfavourable character of oedema and reticulin lesions: and the unconcerned character of cellularity, and lymphocytic and plasmocytic infiltration. These data must be taken into account in an allograft decision.


Isolation of a high-molecular-weight high-mobility-group-type non-histone protein from hen oviduct. An organ-specific non-histone protein, with a mol.wt. of 95,000, was isolated from hen oviduct. This protein consists of approximately equal amounts of acidic and basic amino acids and has an isolectric point of 7.4. On the basis of its known characteristics, this protein is similar to the high-mobility-group proteins observed in other tissues.


The influence of respiratory state on monoamine oxidase activity in rat liver mitochondria. Changes in the respiratory state of rat liver mitochondria caused significant changes (up to 10-fold) in the rates of oxidative deamination of tyramine, indicating interactions between the inner coupling membrane and the monoamine oxidase sites in the outer membrane, and suggesting the possibility that monoamine oxidase is regulated by the thermodynamic state of the mitochondria.


Enhancement by double-stranded polyribonucleotides of production by cultured mouse peritoneal macrophages of differentiation-stimulating factor(s) for mouse myeloid leukaemic cells. Mouse peritoneal macrophages release a factor(s) that stimulates differentiation of a mouse myeloid leukaemic cell line into mature granulocytes and macrophages. Treatment of the macrophages with the synthetic double-stranded polyribonucleotides poly(I).poly(C) and poly(A).poly(U) resulted in enhanced release of the factor into the culture medium. The effect was maximal after treatment with polyribonucleotides for 1 h, and the optimal dose of poly(I).poly(C) was 50 microgram/ml. The single-stranded polyribonucleotides poly(I) and poly(C) at the same concentration were far less effective. The differentiation-stimulating factor was detected not only in the cultured medium but also in the cell lysate. Exposure of macrophages to poly(I).poly(C) enhanced the total activity of the factor in both the culture medium and the cell lysate. The effect of this compound was blocked by the presence of cycloheximide. These results suggest that double-stranded polyribonucleotides enhance production of the differentiation-stimulating factor by peritoneal macrophages.


The reduction of diamide by rat liver mitochondria and the role of glutathione. Diamide is reduced by mitochondria utilizing endogenous substrates with Vmax. 20nmol/min per mg of protein and Km 75micrometer. The reaction is inhibited by: (a) thiol-blocking reagents (N-ethylmaleimide, p-hydroxymercuribenzoate, mersalyl and 2,6-dichlorophenol-indophenol);(b) respiratory inhibitors (arsenicals, malonate and antimycin, but not cyanide or oligomycin; inhibition by antimycin is reversed by ATP); (c) uncouplers (carbonyl cyanide p-trifluoromethoxyphenylhydrazone, 2,4-dinitrophenol and valinomycin with K+; inhibition by the first of these uncouplers is not reversed by cyanide); (d) reagents affecting energy conservation (Ca2+, increasing pH, phosphate; phosphate inhibition is augmented by catalytic ADP or ATP and augmentation is abolished by respiratory inhibitors). Concentrations of mitochondrial glutathione are high when diamide reduction is uninhibited, but low after adding one of the above inhibitors such that the reduction rate is roughly proportional to the glutathione concentration. Endogenous ATP concentrations are lower in the presence of diamide than without, but the difference is abolished by respiratory inhibitors. With oligomycin added, however, ATP concentrations are higher in the presence of diamide and this positive increment is decreased by antimycin, N-ethylmaleimide and malonate. In the presence of diamide and an uncoupler, the mitochondrial glutathione content does not fall if various reducible substrates are present, although the inhibition of diamide reduction is not relieved. Some of these substrates prevent the fall in reduced glutathione concentration found with diamide and phosphate. They also relieve the inhibition of diamide reduction and the relief is sensitive to butylmalonate. The inhibition of diamide reduction by N-ethylmaleimide, mersalyl or p-hydroxymercuribenzoate is not relieved by reducible substrates, but the latter mitigate the fall in the concentration of glutathione. Inhibitors of carriers of tricarboxylic acid-cycle intermediates also inhibit reduction of diamide. The reduced glutathione concentration remains high when they are added singly, but falls when two of them are combined. It is proposed that diamide may enter the matrix as a protonated adduct formed with the thiol groups of mitochondrial carriers and then be reduced in the matrix by glutathione, which is regenerated via NADH, energy-dependent transhydrogenase and NADP+-specific glutathione reductase. Some of the high-energy equivalents required for the transhydrogeneration may be generated by the substrate phosphorylation step of the tricarboxylic acid cycle.


Endocytosis of sulphated proteoglycans by cultured skin fibroblasts. 1. Human skin fibroblasts internalize homologous sulphated proteoglycans by adsorptive endocytosis. Endocytosis rate is half maximal when the concentration of the proteoglycans is 0.1 nM. At saturation, a single fibroblast may endocytose up to 8 X 10(6) proteoglycan molecules/h. 2. The kinetics of prote;glycan binding to the cell surface suggest the presence of 6 X 10(5) high-affinity binding sites per cell. The bulk of sulphated proteoglycans associates to low-affinity binding sites on the cell surface. 3. Glycosaminoglycans and other anionic macromolecules inhibit endocytosis of sulphated proteoglycans non-competitively. The lack of interaction of glycosaminoglycans with the cell-surface receptors for sulphated proteoglycans suggests that the protein core of proteoglycans is essential for binding to the cell surface. 4. The effects of trypsin, cell density, serum concentration and medium pH on endocytosis and degradation of endocytosed sulphated proteoglycans is described. 5. A comparison of the number of the high-affinity binding sites and the number of molecules endocytosed with respect to time suggests a recycling of the proteoglycan receptors between the cell surface and the endocytotic vesicles and/or the lysosomes.


Variation in tissue carnitine concentrations with age and sex in the rat. Diabetes, starvation and various hormonal treatments are known to alter drastically carnitine concentrations in the body. Before the mechanisms controlling carnitine metabolism could be determined, it was necessary to establish normal carnitine concentrations in both sexes at different ages. Carnitine was assayed in plasma, liver, heart and skeletal muscle of rats from birth to weaning. The plasma carnitine increased rapidly during the first 2 days after birth. Carnitine in both heart and skeletal muscle increased, whereas liver concentrations declined during the first week of life. A carnitine-free diet containing sufficient precursors for carnitine biosynthesis was fed to weanling rats. Groups of ten male and ten female rats were killed each week for 10 consecutive weeks. Carnitine was determined in plasma, liver, heart, skeletal muscle, urine and epididymis in the male. There was no difference in carnitine concentrations between the sexes at weaning. Plasma, heart and muscle concentrations were higher in adult male rats than in adult females. However, liver carnitine and urinary carnitine concentrations were higher in adult female than in adult male rats. The epididymal carnitine concentration increased very rapidly during 50 to 70 days of age and the differences in carnitine concentrations between the sexes also became apparent during this time. Thus both the age and the sex of the human subject or experimental animal must be considered when investigating carnitine metabolism.


Age-related changes in the composition and structure of human articular-cartilage proteoglycans. 1. Analysis of the purified proteoglycans extracted from normal human articular cartilage with 4M-guanidinium chloride showed that there was an age-related increase in their content of protein and keratan sulphate. 2. The hydrodynamic size of the dissociated proteoglycans also decreased with advancing age, but there was little change in the proportion that could aggregate. 3. Results suggested that some extracts of aged-human cartilage had an increased content of hyaluronic acid compared with specimens from younger patients. 4. Dissociated proteoglycans, from cartilage of all age groups, bind to hyaluronic acid and form aggregates in direct proportion to the hyaluronic acid concentration. 5. Electrophoretic heterogeneity of the dissociated proteoglycans was demonstrated on polyacrylamide/agarose gels. The number of proteoglycan species observed was also dependent on the age of the patient.


Stable enhancement of calcium retention in mitochondria isolated from rat liver after the administration of glucagon to the intact animal. 1. Mitochondria isolated from rat liver by centrifugation of the homogenate in buffered iso-osmotic sucrose at between 4000 and 8000g-min, 1h after the administration in vivo of 30mug of glucagon/100g body wt., retain Ca(2+) for over 45min after its addition at 100nmol/mg of mitochondrial protein in the presence of 2mm-P(i). In similar experiments, but after the administration of saline (0.9% NaCl) in place of glucagon, Ca(2+) is retained for 6-8min. The ability of glucagon to enhance Ca(2+) retention is completely prevented by co-administration of 4.2mg of puromycin/100g body wt. 2. The resting rate of respiration after Ca(2+) accumulation by mitochondria from glucagon-treated rats remains low by contrast with that from saline-treated rats. Respiration in the latter mitochondria increased markedly after the Ca(2+) accumulation, reflecting the uncoupling action of the ion. 3. Concomitant with the enhanced retention of Ca(2+) and low rates of resting respiration by mitochondria from glucagon-treated rats was an increased ability to retain endogenous adenine nucleotides. 4. An investigation of properties of mitochondria known to influence Ca(2+) transport revealed a significantly higher concentration of adenine nucleotides but not of P(i) in those from glucagon-treated rats. The membrane potential remained unchanged, but the transmembrane pH gradient increased by approx. 10mV, indicating increased alkalinity of the matrix space. 5. Depletion of endogenous adenine nucleotides by P(i) treatment in mitochondria from both glucagon-treated and saline-treated rats led to a marked diminution in ability to retain Ca(2+). The activity of the adenine nucleotide translocase was unaffected by glucagon treatment of rats in vivo. 6. Although the data are consistent with the argument that the Ca(2+)-translocation cycle in rat liver mitochondria is a target for glucagon action in vivo, they do not permit conclusions to be drawn about the molecular mechanisms involved in the glucagon-induced alteration to this cycle.


The use of rat liver nucleoplasm for the characterization of heterogeneous nuclear ribonucleic acid synthesis in vitro. 1. A nucleoplasmic fraction rich in endogenous RNA polymerase II activity was isolated from rat liver nuclei and conditions were determined under which elongation of RNA molecules initiated in vivo continued at maximal rates in vitro. 2. Elongation rates in vitro were calculated to be about 0.25 nucleotide/s and there were about 7 X 10(3) RNA molecules in the process of being elongated by form-II RNA polymerase per original nucleus. 3. Evidence was obtained suggesting that transcription-dependent release of RNA polymerase II molecules from the template occurred during the incubations in vitro. 4. The nascent RNA was tightly associated with protein and banded as ribonucleoprotein in caesium salt gradients. 5. RNA molecules labelled in vitro were up to 13000 nucleotides in length, but consisted of long unlabelled chains transcribed in vivo with only short labelled sequences added in vitro, and without significant polyadenylation. 6. Hybridization of transcripts in the presence of a vast excess of DNA demonstrated that both form-II RNA polymerase and another enzyme, resistant to low alpha-amanitin concentrations, were synthesizing RNA molecules complementary to both reiterated and unique DNA sequences in the genome.


Sources of ammonia for mammalian urea synthesis. The initial rate of incorporation of [15N]alanine into the 6-amino group of the adenine nucleotides in rat hepatocytes was about one-eighteenth of the rate of incorporation into urea. Thus the purine nucleotide cycle cannot provide most of the ammonia needed in urea synthesis for the carbamoyl phosphate synthase reaction (EC 2.7.2.5). On the other hand, contrary to the view expressed by McGivan & Chappell [(1975) FEBS Lett. 52, 1--7], the experiments support the view that hepatic glutamate dehydrogenase can supply the required ammonia.


Effects of polyamines on prostatic chromatin- and non-histone-protein-associated protein kinase reactions. Studies are presented on the influence of polyamines on prostatic chromatin- and non-histone-protein-associated protein kinase reactions involving both exogenous and endogenous substrates. The activities toward the model acidic protein substrate, dephosphophosvitin, were maximal at 160--200mM-NaCl (or -KCl or -NH4Cl). Under these conditions, spermidine and spermine added in concentrations up to 2mM were essentially without effect. However, without addition of NaCl to the medium, marked stimulation of these reactions was elicited by these polyamines at 1--2mM concentrations. The stimulatory effects were not due to non-specific changes in the ionic strength or to substitution of spermine for Mg2+, as maximal stimulation by 1 mM-spermine was observed only at optimal (2--4mM) Mg2+ concentrations. Qualitatively similar effects of polyamines were observed with enzyme preparations from the prostates of castrated rats, and with chromatin and non-histone-protein preparations from other tissues besides ventral prostate. When phosphorylation of endogenous non-histone proteins of the chromatin was measured, spermine stimulated both the initial rates and the final extent of transphosphorylation, even in the presence of optimal concentration of NaCl. By contrast, spermine or spermidine had no effect on the chromatin- and non-histone-protein-associated protein kinase reactions determined with lysine-rich histones as substrates. Chemically NN-dimethylated dephosphophosvitin was a less active substrate for the chromatin-associated protein kinase, but its phosphorylation was more markedly stimulated by spermine in comparison with unmodified dephosphophosvitin. These observations hint that the polyamine stimulations of the various protein kinase reactions may be due to effects on the conformations of the non-histone protein substrates rather than on the kinases themselves.


The development of ketogenesis at birth in the rat. In the suckling newborn rat, blood ketone bodies begin to increase slowly 4h after birth and then rise sharply between 12 and 16h, whereas the major increase in plasma non-esterified fatty acids and liver carnitine occurs during the first 2h of life, parallel with the onset of suckling. In the starved newborn rat, which shows no increase in liver carnitine unless it is fed with a carnitine solution, the developmental pattern of the ketogenic capacity (tested by feeding a triacylglycerol emulsion, which increases plasma non-esterified fatty acids by 3-fold) is the same as in the suckling animal. This suggests that the increases in plasma non-esterified fatty acids and liver carnitine seen 2h after birth in the suckling animal are not the predominant factors inducing the switch-on of ketogenesis. Injection of butyrate to starved newborn pups resulted in a pattern of blood ketone bodies which was similar to that found after administration of triacylglycerols, but, at all time points studied, the hyperketonaemia was more pronounced with butyrate. It is suggested that, even if the entry of long-chain fatty acids into the mitochondria is a rate-limiting step, it is not the only factor controlling ketogenesis after birth in the rat. As in the adult rat, there is a reciprocal correlation between the liver glycogen content and the concentration of ketone bodies in the blood.


Control of gluconeogenesis and of enzymes of glycogen metabolism in isolated rat hepatocytes. A parallel study of the effect of phenylephrine and of glucagon. Hepatocytes isolated from the livers of fed rats were used for a comparative study of the effects of phenylephrine, vasopressin and glucagon on gluconeogenesis and on enzymes of glycogen metabolism. When hepatocytes were incubated in the presence of Ca(2+), phenylephrine stimulated gluconeogenesis from pyruvate less than did glucagon, but, in contrast with this hormone, it did not affect the activities of protein kinase and pyruvate kinase, nor the concentration of phosphoenolpyruvate, and it did not decrease the release of (3)H(2)O from [6-(3)H]glucose. The effects of vasopressin were similar to those of phenylephrine. Gluconeogenesis from fructose was also stimulated by phenylephrine and, more markedly, by glucagon at the expense of the conversion of fructose into lactate. Insulin was able to antagonize the stimulatory effect of phenylephrine on gluconeogenesis from pyruvate. When Ca(2+) was removed from the incubation medium, phenylephrine still stimulated gluconeogenesis from pyruvate, but it also caused an activation of protein kinase and an inactivation of pyruvate kinase; accordingly, the concentration of phosphoenolpyruvate was increased, and, in contrast, vasopressin had no effect on all these parameters. The property of phenylephrine to cause the activation of glycogen phosphorylase was decreased by glucose or by the absence of Ca(2+); it was abolished when these two conditions were combined. Glycogen synthase was inactivated by phenylephrine in the presence or the absence of Ca(2+), although presumably by different mechanisms.


Utilization of methylmalonate for the synthesis of branched-chain fatty acids by preparations of chicken liver and sheep adipose tissue. 1. The utilization of methyl[2-14C]malonyl-CoA for fatty acid synthesis was investigated using synthetase preparations from chicken liver and sheep adipose tissue. 2. The rate of fatty acid synthesis from acetyl-CoA and malonyl-CoA was greatly diminished in the presence of methylmalonyl-CoA. 3. In the absence of malonyl-CoA, methylmalonyl-CoA was utilized for fatty acid synthesis only very slowly by the synthetase from sheep adipose tissue and not at all by that from chicken liver. 4. Despite the inhibitory effect of methylmalonyl-CoA on fatty acid synthesis from malonyl-CoA, it was utilized by the synthetase preparations from both species to produce a complex mixture of methyl-branched fatty acids.


Differential effects of tryptophan on glucose synthesis in rats and guinea pigs. 1. Tryptophan inhibition of gluconeogenesis in isolated rat liver cells is characterized by a 20 min lag period before linear rates of glucose output are attained. 2. Half-maximal inhibition of gluconeogenesis in isolated rat hepatocytes is produced by approx. 0.1 mM-tryptophan. 3. Tryptophan inhibits gluconeogenesis from all substrates giving rise to oxaloacetate, but stimulates glycerol-fuelled glucose production. 4. Gluconeogenesis in guinea-pig hepatocytes is insensitive to tryptophan. 5. Changes in metabolite concentrations in rat liver cells are consistent with a locus of inhibition at the step catalysed by phosphoenolpyruvate carboxykinase. 6. Inhibition of gluconeogenesis persists in cells from rats pretreated with tryptophan in vivo. 7. Tryptophan has no effect on urea production from alanine, but decreases [1-14C]palmitate oxidation to 14CO2 and is associated with an increased [hydroxybutyrate]/[acetoacetate] ratio. 8. These results are discussed with reference to the control of gluconeogenesis in various species.


Characteristics of the transport of alanine, serine and glutamine across the plasma membrane of isolated rat liver cells. 1. Alanine, glutamine and serine were actively accumulated in liver cells isolated from starved rats. 2. This accumulation was inhibited when either Na+ or HCO3- ions were omitted from the incubation medium. In general the degree of dependence on Na+ was quantitatively similar to that on HCO3-. 3. The apparent Km values for the transport of all three amino acids were in the range 3--5mM with Vmax. values in the range 15--25nmol/min per mg of cell protein at 37 degrees C. 4. Alanine and serine transport were mutually competitive; glutamine inhibited the transport of alanine and serine non-competitively. 5. The initial rate of transport of these amino acids was inhibited when the intracellular content of ATP was decreased. 6. Ouabain inhibited the rate of alanine transport without inhibiting the rate of alanine metabolism. 7. It is concluded that a minimum of three transport systems must be postulated to exist in the liver cell plasma membrane to account for the transport of alanine, serine and glutamine. The rate of transport of these amino acids in isolated hepatocytes is unlikely to limit the rate at which they are metabolized.


The effects of ammonium chloride and bicarbonate on the activity of glutaminase in isolated liver mitochondria. 1. Glutamine hydrolysis in liver mitochondria was studied by measuring the production of glutamate under conditions where this compound could not be further metabolized. 2. Glutaminase activity in intact mitochondria was very low in the absence of activators. 3. Glutamine hydrolysis was markedly stimulated by NH4Cl and also by HCO3- ions. 4. The stimulation by each of these compounds was much decreased if the mitochondria were uncoupled. 5. Maximum rates of glutamine hydrolysis required the addition of phosphate. A correlation was observed between the activity of glutaminase in the presence of NH4Cl plus HCO3- and the intramitochondrial content of ATP. 6. In disrupted mitochondria, NH4Cl stimulated glutaminase to a much smaller extent than in intact mitochondria. The NH4Cl stimulation in disrupted mitochondria was much increased by the addition of ATP. KHCO3 also stimulated glutaminase activity in disrupted mitochondria, and ATP increased the magnitude of this stimulation. 7. It was concluded that maximum rates of glutaminase activity in liver mitochondria require the presence of phosphate, ATP and either HCO3- or NH4+. A comparison of the results obtained on intact and broken mitochondria indicates that these effectors have a direct effect on the glutaminase enzyme system rather than an indirect effect mediated by changes in transmembrane ion gradients or in the concentrations of intramitochondrial metabolites.


Post-translational regulation of lipoprotein lipase activity in adipose tissue. Changes in adipose-tissue lipoprotein lipase activity that are independent of protein synthesis were investigated in an incubation system in vitro. Under appropriate conditions at 25 degrees C a progressive increase in the enzyme activity occurs that is energy-dependent. Part of the enzyme is rapidly inactivated when the tissue is incubated with adrenaline or adrenaline plus theophylline. The mechanism of this inactivation appears to be distinct from, and to follow, the activation of the enzyme. A hypothesis is presented to account for the results in terms of an activation of the enzyme during obligatory post-translational processing and a catecholamine-regulated inactivation of the enzyme as an alternative to secretion from the adipocyte.


The binding of 3H-labelled oestradiol- and progesterone-receptor complexes to hypothalamic chromatin of male and female sheep. Chromatin isolated from hypothalamic nuclei of sexually mature entire male and female sheep was linked to cellulose in u.v. light. The saturation binding of 3H-labelled oestrogen- and progesterone-receptor complexes, prepared by (NH4)2SO4 precipitation from the 105000g supernatant of hypothalamic cytosol, was then measured in vitro in 0.15m-KCl. Saturation binding was also measured after extraction of histones and masking acidic proteins. Salt + urea was observed to be more effective than guanidine hydrochloride in unmasking receptor acceptor sites, and the binding of labelled receptor complexes to dehistonized unmasked chromatin was shown to be largely resistant to 0.4m-KCl extraction. Whereas extents of receptor-complex binding were similar to published values for comparable preparations of hen oviduct chromatin, no sex-related difference was observed. However, binding of progesterone-receptor to chromatin was greater than that of oestradiol-receptor. Binding also increased more after removal of histones and masking acidic proteins, suggesting the presence of a greater number of progesterone-receptor acceptor sites in hypothalamic chromatin than of estradiol-receptor acceptor sites. The failure to demonstrate a sex-related difference in oestradiol-receptor binding to hypothalamic chromatin in vitro is discussed.


Stimulation by vasopressin, angiotensin and oxytocin of gluconeogenesis in hepatocyte suspensions. 1. In hepatocytes from starved rats, vasopressin, angiotensin (angiotensin II) and oxytocin stimulated gluconeogenesis from lactate by 25--50%; minimal effective concentrations were about 0.02pM, 1 nM and 0.2 nM respectively. 2. Vasopressin and angiotensin also stimulated gluconeogenesis from alanine, pyruvate, serine and glycerol. EGTA decreased gluconeogenesis from these substrates. 3. Hormonal stimulation of gluconeogenesis from lactate was abolished in the absence of extracellular Ca2+. 4. Insulin did not prevent stimulation of gluconeogenesis by vasopressin or angiotensin. 5. The potency of the stimulatory effects of vasopressin and angiotensin on hepatic gluconeogenesis suggests they are operative in vivo. Also, the data suggest that Ca2+ plays a role in the stimulation by these hormones.


Studies of a factor from dystrophic mouse muscle inhibitory towards protein synthesis. A substance inhibitory to protein synthesis was purified from mouse skeletal muscle by gel filtration and ion-exchange chromatography, as well as by centrifugation on sucrose gradients. The molecular weight of the inhibitor, determined by sodium dodecyl sulphate/polyacrylamide-gel electrophoresis, was 71000. The inhibitory activity was insensitive to ribonuclease A, deoxyribonuclease I and phospholipase C. It was sensitive to Pronase treatment but insensitive to heat-treatment and trypsin degradation. The present results, taken together with previous studies, indicate that the site of action of the inhibitor is not on the initiation phase of protein synthesis but rather at a step after the binding of aminoacyl-tRNA to ribosomes. The increased inhibitor activity found in dystrophic muscle is discussed.


Choice of precursors for the measurement of protein turnover by the double-isotope method. Application to the study of mitochondrial proteins. 1. The double-isotope concept [Arias, Doyle & Schimke (1969) J. Biol. Chem. 224, 3303--3315] for the measurement of protein turnover was used to estimate the turnover of proteins in subcellular and submitochondrial fractions prepared from rat liver. 2. Double-isotope experiments with [3H]leucine as first precursor and [14C]leucine as second precursor were used to measure the turnover rates of proteins in subcellular and submitochondrial fractions. Solvent extraction procedures designed to remove lipids and nucleic acids from trichloroacetic acid precipitates only changed the isotope ratio of the microsomal fraction. It was not possible to measure turnover of proteins in mitochondrial and submitochondrial fractions with these precursors. 3. Double-isotope experiments were designed to minimize first-precursor reutilization by employing NaH14CO3. [3H]Arginine was used as second precursor. The turnover rates of protein in subcellular and submitochondrial fractions was measured. Solvent extraction procedures designed to remove lipids and nucleic acids showed changes in the isotope ratio for all subcellular fractions, especially in microsomal and detergent-soluble mitochondrial fractions. Isotope ratios of precipitates after solvent extraction indicate that, whereas considerable heterogeneity exists for the average rates of protein turnover in subcellular fractions, little heterogeneity is observed in the average rates of protein turnover in submitochondrial fractions.


Onset of ribosome degradation during cessation of growth in BHK-21/C13 cells. When BHK-21/C13 cells growing exponentially in 10% serum are transferred to a medium containing only 0.25% serum, cell growth is decreased. After initial changes in RNA synthesis and degradation, protein content of the cultures reaches a plateau and eventually DNA synthesis is arrested. rRNA is relatively stable in exponentially growing cells. Immediately after 'step-down' rRNA degradation commences, but poly(A)-containing RNA does not appear to be degraded any faster than in control cells. Reutilization of RNA precursors has been independently measured and amounts to less than 1%/h for rRNA, insufficient to influence the conclusion that rRNA degradation begins almost immediately after 'step-down'. The degree of reutilization of uridine is much greater for poly(A)-containing RNA than for poly(A)-free RNA.


An alternative hypothesis of cellular transport of lysosomal enzymes in fibroblasts. Effect of inhibitors of lysosomal enzyme endocytosis on intra- and extra-cellular lysosomal enzyme activities. Recapture of lysosomal enzymes secreted by fibroblasts was inhibited by growing the cells in the presence of either free or immobilized antibodies against lysosomal enzymes or in the presence of phosphorylated carbohydrates known to interact with the cell-surface receptors for lysosomal enzymes. The following results were obtained. 1. Conditions that prevent recapture of released lysosomal enzymes increase the rate of extracellular accumulation of these enzymes up to twice that of controls. 2. Growing cells for 12 days in the presence of 0.5mm-mannose 6-phosphate, which decreases beta-N-acetylglucosaminidase endocytosis to less than 10% of that of controls, has no effect on the intracellular activity of this and four other lysosomal enzymes. 3. Growing cells for 4 days in the presence of 50mm-mannose 6-phosphate, which is a 1000-fold higher concentration than that required for 50% inhibition of lysosomal enzyme endocytosis, leads to a 4-fold increase in extracellular beta-N-acetylglucosaminidase accumulation and a decrease in intracellular enzyme. These results give evidence that, in fibroblasts, transfer of lysosomal enzymes into lysosomes does not require secretion before a receptor-mediated recapture [Hickman & Neufeld (1972) Biochem. Biophys. Res. Commun.49, 992-999]. We propose that (a) lysosomal enzymes are present in a receptor-bound form in those vesicles that fuse with the cell membrane, (b) the major part of the lysosomal enzyme cycles via the cell surface in a receptor-bound form and (c) only a minor part of the lysosomal enzyme is released into the extracellular space during its life cycle.


Metabolism of inorganic sulphate in the isolated perfused rat liver. Effect of sulphate concentration on the rate of sulphation by phenol sulphotransferase. 1. The metabolism of inorganic [(35)S]sulphate (Na(2) (35)SO(4)) was studied in the isolated perfused rat liver at three initial concentrations of inorganic sulphate in the perfusion medium (0, 0.65 and 1.30mm), in relation to sulphation and glucuronidation of a phenolic drug, harmol (7-hydroxy-1-methyl-9H-pyrido[3,4-b]indole). 2. [(35)S]Sulphate rapidly equilibrated with endogenous sulphate in the liver. It was excreted in bile and reached, at the lowest concentration in the perfusion medium, concentrations in bile that were much higher than those in the perfusion medium; at the higher sulphate concentrations, these concentrations were equal. The physiological concentration of inorganic sulphate in the liver, available for sulphation of drugs, is similar to the plasma concentration. 3. At zero initial inorganic sulphate in the perfusion medium, the rate of sulphation was very low and harmol was mainly glucuronidated. At 0.65mm-sulphate glucuronidation was much decreased and considerable sulphation took place, indicating efficient competition of conjugation by sulphation. At 1.30mm-sulphate the sulphation increased still further. 4. The results suggest that an important factor in sulphation is the relatively high K(m) of synthesis of adenosine 3'-phosphate 5'-sulphatophosphate (the co-substrate of sulphation) for inorganic sulphate, which is of the order of the plasma concentration of inorganic sulphate. The steady-state adenosine 3'-phosphate 5'-sulphatophosphate concentration may determine the rate of sulphate conjugation of drugs in the rat in vivo.


Conversion between renin and high-molecular-weight renin in the dog. Two forms of renin, one of mol.wt. 43,000 and the other 60,000, were found in the dog kidney. Conversion between the two forms of renin was reversible at neutral pH. Though the molecular weight of renin in kidney-cortex homogenate was 43,000, it was completely converted into high-molecular-weight renin in the presence of substances that react with thiol groups. On the contrary, stored renin in the granules was the form of normal size (mol. wt. 43,000) regardless of the absence or presence of such substances. The present experiments indicated that renin is stored in the granules as the form of normal size and might be converted into high-molecular-weight renin when it is released from the granules and attached to some substance in the soluble fraction of renal-cortical tissue.


Anion/calcium ion ratios and proton production in some mitochondrial calcium ion uptakes. The uptake of Ca2+ by liver mitochondria, when phosphate movement is inhibited, occurs when Co2 is present and not in its absence. Uptake of Ca2+ to form CaCO3 yields 2H+/Ca2+. Heart mitochondria, when phosphate movement is inhibited, will take up Ca2+ with the exact equivalent of hydroxybutyrate, lactate or acetate. By providing a carrier for Cl- with tributyltin, a stoicheiometric uptake of Cl- with the Ca2+ takes place. The uptakes appear to occur without significant pH change; there appears to be no CO2-dependent uptake into heart mitochondria. Oxygenation of anaerobic heart mitochondria, in the presence of an inhibitor of phosphate movement and of generation of phosphate from internal ATP, does not yield significant change of external acidity in relation to the amount of O2 added. Use of Bromothymol Blue as an indicator of the distribution of a weak acid anion confirms that the transient nature of the response of the dye distribution to Ca2+ is connected with movement of endogenous phosphate. Bromothymol Blue accumulated in response to Ca2+ is discharged when entry of the Ca2+ (in the presence of mersalyl) is mediated with nigericin. It is concluded that Ca2+ uptakes will occur alternatively with the equivalent of anions or in exchange for endogenous K+ and that proton production is connected with the changes of ionization of phosphate (unless phosphate movement is inhibited) and in liver mitochondria with the hydration of CO2.