[Effect of hyperchylomicronemia and hyperprebetalipoproteinemia on the vascular walls of rats of different ages]. The effect of hyperchylomicronemia and hyperprebetalipo-proteinemia on the vascular wall of 6--10 and 26--30-month-old rats was studied. These animals were injected intravenously with the blood serum taken from rats, preliminarily treated with Triton WR 1339 (intraperitoneal administration). The electron microscopy study revealed the presence of inclusion of chylomicrons and lipoproteins of very low density in the endothelium of old animals only. Swelling of the Golgi complex and mitochondria, expansion of the cysterns of endoplasmic reticulum in the endothelial cells of young rats pointed to the activation of intracellular metabolic processes. The study under these conditions of the biosynthesis of some lipid classes in the aorta showed a similar inhibition of free cholesterol biosynthesis in rats of various age. An increase of phospholipid biosynthesis was recorded in young animals only.


[Electrophoretic study of the nuclear membrane ribonucleases of rat liver and hepatoma-27 cells]. Purified cell nuclei from the rat liver and hepatoma-27 cells were used to prepare nuclear membranes from which the enzyme-containing extracts of acid-soluble proteins were prepared. The protein extracts were subjected to disc-electrophoresis in 15% polyacrylamide gel using modified Reisfeld's system. It was found that ribonucleases contained in the acid-soluble proteins of the nuclear membranes of normal liver are represented as several components, and differed by their electrophoretic mobility and also by some other physical and chemical properties from crystalline bovine ribonuclease, as well as from nuclear chromatine ribonucleases.


[Effect of cholesterol on membrane Na,K-activated adenosine triphosphatase]. The membrane preparations of rat brain Na,K-ATP-ase with different amount of cholesterol were obtained by using high-cholesterol diet or cholesterol-oxidase treatment. The cholesterol/protein ratio varied from 0.034 to 0.252 (normal--0.152). An increase of this ratio removes the curvature on the Arrhanius' plots for Na,K-ATP-ase, and a decrease--enhances it. The change of the temperature dependence pattern on the enzyme activity correlates with that of the spectral parameter of spin-labelled and rostane analogue, characterizing fluidity of membrane lipids in the same preparations. The authors conclude that the cholesterol level in the brain membranes monitors the activity of the membrane Na,K-ATP-ase.


[Mechanisms of the different effect of sucrose on the quantity of lipids in the blood and liver of experimental animals]. Experiments on rats showed that inclusion of sucrose into the composition of diets with increased or normal carbohydrate content accelerated the synthesis in the liver of apoproteins of pre-beta-lipoproteins and their loading with endogenously formed lipids, promoted a more intensive secretion of lipoproteins of very low density into the blood and an increase in it of the triglyceride level. However, the rate of the lipid compensation with the protein of pre-beta-lipoproteins in the liver depends also on the qualitative ratio of the carbohydrate and fat components in the diet. In using the diet with sucrose with the physiological ratio of carbohydrates and fats (2:1) there was noted a lower load of apoproteins of pre-beta-lipoproteins with lipids, this leading to an increase of the latter in the liver.


[Activity of glucose-6-phosphate metabolism enzymes in the livers of rats with experimental valekson poisoning]. The activity of hexokinase, glucose-6-phosphatase and glucose-6-phosphoric dehydrogenase was studied in the liver of rats after one hour, one and five days after a single oral administration of organic phosphorus insecticide valekson. It was determined that administration of the preparation led to an increase of activity in the homogenate and solubilization of glucose-6-phosphatase, activation of glucose-6-phosphoric dehydrogenase and inhibition of hexokinase. The changes were maximum one hour after the administration of the compound. The results show that a decrease of the intensity of glucose-6-phosphate formation and metabolism is one of the pathogenetic factors in the development of valekson-induced intoxication.


[Immunogenicity and thymus-dependence of the polymerized alpha-toxoid of Clostridium perfringens]. Polymeric alpha-toxoid with a molecular weight of 450 000--600 000 was obtained by condensation of alpha-toxoid of Cl. perfringens, type A, with glutaric aldehyde. Experiments on guinea pigs showed that in the adsorbed preparations the immunogenic properties of both monomeric and polymeric alpha-toxoids are practically identical. The primary immune response after the immunization with nonadsorbed antigens was 3 times greater than that induced by the monomer. Polymerization of alpha-toxoid failed to change its thymus dependency.


The mechanism of action of narcotic analgesics in the guinea-pig ileum. 1. Intracellular recordings were made from neurones in the myenteric plexus of the guinea-pig ileum. Single myenteric ganglia were maintained in vitro and drugs were applied by adding them to the perfusing solution. 2. Narcotic analgesics hyperpolarized the membrane of a proportion of neurones in the myenteric plexus. 3. The membrane hyperpolarization was sometimes associated with a decrease in input resistance. These effects reduced the excitability of myenteric neurones. 4. The effects of narcotics occurred at low concentrations (10 nM to 1 micrometer), were stereospecific and were reversed by naloxone. 5. It is proposed that the morphine-sensitive neurones may be the cholinergic efferents to the muscle layers. By hyperpolarizing these neurones, morphine may prevent their excitation by electric field stimulation. This may explain why narcotic analgesics reduce the output of acetylcholine and the contractile response of this preparation when it is excited by field stimulation.


Relaxation of smooth muscle following contraction elicited by sympathetic nerve stimulation in vivo. 1. A method for studying in vivo the process of neuroeffector transmission in the nictitating membrane and nasal blood vessels of the cat is described. 2. Administration of desmethylimipramine or cocaine caused increases in both the amplitude and duration of the nasal and membrane responses which may be explained by inhibition of neuronal uptake of noradrenaline. 3. Phenoxybenzamine depressed the responses to nerve stimulation, but had little effect on the relationship between response amplitude and rate of recovery. 4. The relationship between response amplitude and rate of recovery is discussed and related to the sigmoid shape of a log concentration-response curve.


Kinin, kininogen and kininase levels during acute Babesia bovis (= B. argentina) infection of cattle. 1. Kinin levels began rising on day 3 after infection of cattle with Babesia bovis (= B. argentina) and attained a maximum value of 98% above preinfection levels by day 7. 2. Kininogen levels began falling on day 3 and reached minimum levels of 83% below preinfection levels on day 8. 3. Changes in both kinin and kininogen levels on day 3 coincided with the detection of low levels of parasites, and with a fall in packed cell volume. 4. Plasma kininase levels rose significantly 6 to 9 days after infection. Preparations of lysed and sonicated uninfected and infected red cells contained kininase activity, the respective red cell preparations being 23.9 and 11.4 times more active per mg protein than uninfected red cell preparations. The effect of pH, and the inhibitors disodium edetate, 1,10 phenanthroline and aprotinin on normal and infected plasma and on the various red cell preparations suggested that the rise in plasma kininase levels during infection was probably at least partly due to parasite products. 5. These results are discussed in relation to previous data showing that both kallikrein activation and the onset of hypotension also occur on or about day 3.


The mode of action at the mouse neuromuscular junction of the phospholipase A-crotapotin complex isolated from venom of the South American rattlesnake. 1 Phospholipase A(2)-crotapotin complex (P-C complex) isolated from the venom of Crotalus durissus terrificus induced an irreversible blockade of neuromuscular transmission when twitch tension was measured in the mouse phrenic nerve-hemidiaphragm preparation in vitro at 37 degrees C.2 A similar concentration of the phospholipase A(2) (10 mug/ml) alone did not affect neuromuscular transmission and no priming action was detected on later addition of crotapotin.3 The rate of neuromuscular blockade induced by P-C complex (15 mug/ml) was not altered by raising the frequency of nerve stimulation. Lower temperatures markedly increased the time of onset and reduced the rate of blockade (Q(10) (27-37 degrees C) of 4.4) whilst replacement of Ca by Sr in the medium prevented this activity. These latter results suggest that enzymatic activity is important in the neurotoxicity of the complex.4 A myotoxic action was shown by 30 mug/ml P-C complex and 30 mug/ml phospholipase A(2).5 P-C complex (150 mug) was injected into the tail vein of mice and the intoxicated hemidiaphragm preparation removed for intracellular recording at 25 degrees C.6 In fully intoxicated hemidiaphragms, resting membrane potentials were unaltered and endplate potentials (e.p.ps) varied in average amplitude from zero to less than 3 mV.7 Miniature endplate potential (m.e.p.p.) frequency was lower at fully poisoned endplates than at controls; the frequency rose during a 50 Hz tetanus but was unaffected by either raising external K or the application of the Ca-ionophore A23187.8 E.p.ps were recorded in partially intoxicated hemidiaphragms with (+)-tubocurarine (0.5-1.0 mug/ml) added to prevent contraction. Evoked release was abnormal as 50 Hz tetanus elicited e.p.ps of very variable amplitude, no facilitation of response was shown to paired stimuli, and tetraethylammonium (0.5 mM) failed to increase e.p.p. amplitudes.9 M.e.p.ps and e.p.ps were recorded at partially poisoned endplates in low Ca-high Mg solution. A reduction in the quantal content of evoked transmitter release was observed in comparison with controls.10 M.e.p.ps recorded at partially and at fully intoxicated endplates showed an altered amplitude distribution with a higher proportion of large potentials.11 It is concluded that P-C complex has a presynaptic site of action and may interfere with depolarization-secretion coupling at the motor nerve terminals.


The action of melatonin on single amphibian pigment cells in tissue culture. 1. Neural crest material from neurula stage Xenopus embryos was tissue cultured as small aggregates of cells or a single cell suspension. Isolated pigment cells differentiated after 2 days in culture. 2. Melatonin (10(-15) to 10(-13) M) induced pigment granule condensation; it was 10,000 times more effective than any other compound tested. 3. Tests with appropriate agents showed the pigment cells to have beta-adrenoceptors and 5-hydroxytryptamine receptors; these sites could be blocked without affecting the response to melatonin. Phentolamine blocked the effect of melatonin. 4. Removal of Na+ or Ca2+ from the bathing medium inhibited melatonin-induced pigment granule condensation; 10 mM K+ induced pigment granule condensation. D600, Mn2+ and La3+, which inhibit Ca2+ entry, blocked the effect of melatonin. 5. Cyclic GMP induced pigment condensation and cyclic AMP pigment dispersion (10(-2) to 10(-4) M). 6. It is suggested that the action of melatonin is accompanied by depolarization and the entry of calcium ions. 7. Pigment cells in tissue culture could provide a useful bioassay method for melatonin.


An inhibitory role for noradrenaline in the mouse vas deferens. 1 Noradrenaline (0.1-3.0 muM) inhibited the twitch responses to single pulse field stimulation of the isolated vas deferens of the mouse. The higher concentrations of noradrenaline (ca. 0.3-3.0 muM) were required to make the tissue contract.2 Phentolamine (10 muM) abolished the contractor response to higher concentrations of noradrenaline and antagonized the inhibitory effect of lower concentrations on the twitch response.3 Propranolol (10 muM) potentiated both the contractor and the inhibitory effect of noradrenaline on the twitch response.4 Isoprenaline (0.1-3.0 muM) and salbutamol (1.0-3.0 muM) both inhibited the twitch response. Their effects were antagonized by propranolol (10 muM), but not by practolol (10 muM).5 The effects of uptake(1) and uptake(2) blocking agents were determined. Cocaine (10 muM) reduced the size of the twitch response in 2 out of 4 experiments. Imipramine (0.18 muM) also reduced the size of the twitch, as did oestradiol (3.7 muM) and a combination of cocaine and oestradiol.6 Contractor responses to exogenous noradrenaline showed tachyphylaxis, but when this was not very marked, the response could be shown to be potentiated by uptake blocking agents.7 The inhibitory effect of noradrenaline on the twitch response was greatly potentiated by cocaine (10 muM) and much less so by oestradiol (3.7 muM).8 It is concluded that the transmitter responsible for the twitch response is either an unknown substance released from the sympathetic neurone, or noradrenaline acting upon a receptor with none of the characteristics of known alpha- or beta-adrenoceptors. In either case, noradrenaline can inhibit the output, probably by stimulation of presynaptic alpha-adrenoceptors.


Diuretics and the renal adenylate cyclase system. 1 The relationship between the diuretic effectiveness and the effect on the renal adenylate cyclase of three diuretics, acetazolamide, frusemide and ethacrynic acid, was examined. The hypothesis that acetazolamide and parathyroid hormone (PTH), inhibit renal carbonic anhydrase by a cyclic adenosine 3',5'-monophosphate (cyclic AMP)-dependent mechanism was also tested.2In vitro, acetazolamide, frusemide and ethacrynic acid at high concentrations (10(-3)M) all produced some inhibition of basal and stimulated rat kidney plasma membrane adenylate cyclase. The effect of acetazolamide was much less than that of frusemide and ethacrynic acid. These plasma membrane effects were reproduced in studies of cyclic AMP formation in isolated kidney tubules of rats.3 Intravenous injections of acetazolamide did not change the total cyclic AMP content of the kidneys of rats killed by microwave irradiation.4 Acetazolamide produced a diuresis in the rat and a slight inhibition of the antidiuretic effect of Pitressin. Frusemide produced a diuresis and greatly reduced the antidiuretic response to Pitressin. Ethacrynic acid was ineffective as a diuretic in the rat and actually enhanced the antidiuretic response to Pitressin.5 In investigating the possible influence of diuretics and PTH on the activity and state of phosphorylation of carbonic anhydrase it was found that: there was no correlation between the ability of diuretics to inhibit carbonic anhydrase activity and to inhibit carbonic anhydrase phosphorylation; neither PTH nor cyclic AMP (in the presence of adenosine triphosphate, Mg(2+), K(+) and incubation at 37 degrees C) inhibited rat cortex homogenate carbonic anhydrase activity.6 It seems unlikely that any of the tested diuretics exerts its pharmacological effect by means of changes in kidney cyclic AMP metabolism.


An estimate of the equilibrium dissociation constant for calcium as an antagonist of evoked acetylcholine release: implications for excitation-secretion coupling. The equilibrium dissociation constant (K(d)) for Ca(2+) as an antagonist of evoked acetylcholine (ACh) release was determined in the hope of distinguishing whether divalent cations control excitation-secretion coupling selectively (by binding with high affinity to an external membrane site) or non-selectively (by screening fixed negative charges on the external surface of the nerve terminal). ACh release was detected electrophysiologically by means of conventional intracellular recording techniques at frog motor endplates. Ba(2+) was used as the agonist to support the asynchronous release of ACh by repetitive motor nerve impulses. Despite its dispersed nature, release mediated by Ba(2+) occurs through the same conductance pathway as synchronous release mediated by Ca(2+). Ca(2+) was found to be a potent antagonist of Ba(2+)-dependent release with a K(d)=0.12+/-0.02 mM (mean+/-s.e. mean, n=5). This value is 30-50 times lower than the K(d) for Mg(2+) as an antagonist of the same release process. It is suggested that antagonism of release by Ca(2+) is likely to be exerted at the same external site that binds other divalent cation antagonists, a site that appears essential for the agonist behaviour of Ca(2+). The high affinity (low K(d)) of Ca(2+) as an antagonist of ACh release suggests that a selective, binding model appears to be the most appropriate single description of the action of divalent cations at the external surface of the motor nerve ending.


beta-Adrenoceptor stimulation of exocrine secretion from the rat pancreas. 1 Effects of catecholamines given intravenously on exocrine secretion from the pancreas were investigated in anaesthetized rats. The flow rate of pancreatic juice under resting conditions was 11.1 +/- 3.2 microliter per hour in 100 animals. 2 Dopamine (0.3--3 mg/kg) and isoprenaline (1--10 microgram/kg) induced almost the same increase in the pancreatic secretion, so that dopamine was 300 times less potent than isoprenaline. The relative potency of the two amines for stimulation of pancreatic secretion was equivalent to that for beta-stimulation of the contractile force of the left ventricle in vivo. 3 Propranolol (0.5 mg/kg) antagonized completely the dopamine- and isoprenaline-induced stimulation of the pancreatic secretion. 4 Haloperidol (10 mg/kg) failed to suppress the secretory effect of dopamine on the exocrine pancreas but abolished the dopamine-induced hypotension. 5 The dopamine-induced secretion was not modified by atropine (3 mg/kg), phenoxybenzamine (3 mg/kg), vagotomy or pithing. 6 Adrenaline and noradrenaline (10 microgram/kg) induced secretion after phenoxybenzamine  treatment (3 mg/kg). 7 It is suggested that the rat pancreas has a stimulatory beta-adrenoceptor mechanism of exocrine secretion.


The influence of prostaglandins on noradrenaline-induced vasoconstriction isolated perfused mesenteric blood vessels of the rat. 1 The report of the depression by indomethacin of vasoconstrictor responses to noradrenaline and their partial restoration by prostaglandin E(2) (PGE(2)) and PGE(1) in rat isolated perfused mesenteric blood vessels was investigated. The further suggestion that prostaglandins may be necessary for the combination of noradrenaline with the alpha-adrenoceptor in this tissue was also studied.2 The reported depression by indomethacin was confirmed and was further shown to be in the form of a concentration-dependent flattening of the noradrenaline concentration-effect curve.3 A concentration-dependent restorative effect was observed for all prostaglandins studied. The decreasing order of potency for the restoration towards normal of the indomethacin-depressed responses to noradrenaline was: PGE(2), PGE(1), PGA(1), PGF(2alpha), PGA(2).4 The prostaglandins studied were not uniform in their restorative actions and could be separated into two groups. PGE(2) and PGE(1) restored responses towards the control level whereas PGA(1), PGA(2) and PGF(2alpha) increased responses to an above control level and did so over a smaller concentration range. The possibility of several prostaglandin receptors is discussed.5 At concentrations equi-effective in restoring depressed responses to control levels PGA(1) but not PGE(2), caused a parallel shift of the noradrenaline concentration-effect curve to the left and a small, gradual rise in the basal perfusion pressure.6 The reason for the differing effects remains obscure but does not seem to involve a change in the alpha-adrenoceptor as indicated by the pA(2) of phentolamine. Furthermore, the restorative and potentiating effect of PGA(1) is not mediated by blockade of neuronal uptake of noradrenaline.7 It appears that prostaglandins are required for the vasoconstrictor action of noradrenaline in rat mesenteric blood vessels and that this effect is distal to the drug-receptor interaction. The possible involvement of prostaglandins with intracellular calcium ions is discussed.


Immune status of children of immigrants to poliomyelitis. Four ethnic groups of children from the Glasgow area--155 Asians, 85 Africans, 85 Chinese, and 93 Scots--were examined for neutralising to poliovirus types 1, 2, and 3. Only seven of the 418 children had no detectable antibody, and of these, four were aged less than 7 months; none had received polio vaccine. The best-protected children were the Chinese (93% with antibody to all three poliovirus types), followed by the African (81%), Scottish (78%), and Asian children (77%). We conclude that children of immigrants are no more vulnerable to poliovirus infection than their Scottish counterparts.


Methods for the study of the glycoproteins and proteoglycans of bone using bacterial collagenase. Determination of bone sialoprotein and chondroitin sulphate. A method has been developed for the solubilization of bone matrix using bacterial collagenase. The soluble macromolecules were separated by a column procedure on DEAE-cellulose and analysed for uronic and sialic acids. Values for the total non-collagenous fraction of bone and its content of bone sialoprotein and chondroitin sulphate were obtained. The method was compared with similar procedures using EDTA extraction and papain digestion.


Absence of mitochondrial terminal respiratory enzymes in cartilage matrix vesicles. This study attempted to detect evidence of mitochondrial terminal respiratory components in matrix vesicles isolated from rachitic rat tibial epiphyseal plates. Biochemical assays for cytochrome c oxidase, NAD isocitrate dehydrogenase, NADP isocitrate dehydrogenase and succinate-cytochrome c reductase were negative. Polarimetric determinations revealed that the addition of succinate to matrix vesicles in suspension did not cause any increase in oxygen utilization. Spectrophotometric tracings of deoxycholate-solubilized matrix vesicles showed no characteristic absorption peaks or maxima belonging to any of the cytochrome complex components. Attempts to prepare pyridine hemochromes of cytochrome prosthetic groups from the matrix vesicles were also unsuccessful. The above results indicate that key components of mitochondrial respiratory systems are not detectable in rachitic matrix vesicles. The results are compatible with the interpretation that such vesicles are not derived from mitochondria.


Chromatographic separation of two acid phosphatases from rat bone. Extracts of tibiae of suckling rats were prepared with 0.3 M KCl containing 0.1% Triton X-100 and were chromatographed with CM-52 cellulose. Most of the acid phosphatase activity determined with p-nitrophenylphosphate (p-NPP) was bound to the cellulose and could be eluted with a sodium acetate buffer gradient in 2 distinct peaks. The major peak, E2, was bound strongly to the cellulose and showed high activity with p-NPP and inorganic pyrophosphate (P-Pi), but only slight activity with beta-glycerophosphate (beta-GP) and was unaffected by tartrate. The minor peak, E1, was weakly bound to the adsorbent, showed equal activity with p-NPP and beta-GP, but negligible activity with P-Pi and was completely inhibited by tartrate. These results support earlier evidence suggesting that bone contains at least 2 different acid phosphatases and that the more abundant enzyme may function as a pyrophosphatase.


An improved procedure for the purification of plasma membranes from Dictyostelium discoideum. A novel procedure was recently described for the purification of plasma membranes of Dictyostelium discoideum (Gilkes, N. R. & Weeks, G. (1977) Biochim. Biophys. Acta 464, 142-156). Considerable enrichment of plasma membrane marker enzymes was achieved, but since purified mitochondrial and endoplasmic reticulum fractions were unavailable, it was not possible to accurately assess the contamination level of these organelles. We have therefore slightly modified the plasma membrane preparation procedure, improving purification, and have prepared partially purified mitochondrial and endoplasmic reticulum fractions. The data suggest that the contamination of the plasma membranes by endoplasmic reticulum membranes is no greater than 10%, and probably considerably less. No mitochondrial contamination is detectable.


Increased response of adipose tissue of the ob/ob mouse to the action of adrenaline after treatment with thyroxin. The activity of the lipolytic system of the obese hyperglycemic mouse was assessed after treatment with physiological doses of thyroxin (T4). The treatment significantly increased fatty acid mobilization in response to adrenaline over the levels observed in the control mice under all conditions studied. The activities of the high- and low-Km phosphodiesterases and of adenylate cyclase were also studied. Treatment of the ob/ob mice with T4 had little effect on the activities of the cyclic AMP phosphodiesterases (high and low Km) but it partially restored the activity of adenylate cyclase, which is deficient in these animals. A correlation was found in the T4-treated obese animals between the ability of the epididymal adipose tissue to mobilize fatty acids, its ability to increase the intracellular levels of cyclic AMP, and the activity of adenylate cyclase in response to adrenaline stimulation.


A convenient method of establishing permanent lines of xeroderma pigmentosum cells. Nine lymphoblastoid cell lines were established after transformation by Epstein-Barr virus of peripheral lymphocytes from four xeroderma pigmentosum (XP) patients, the parents of one XP patient, and three normal donors. All these cell lines proliferate as suspension in Roswell Park Memorial Institute Medium 1640 supplemented with 20% fetal bovine serum, without detectable release of infectious Epstein-Barr virus. Some characteristics of these cell lines, such as growth rates, chromosome numbers, UV sensitivities, and activities of unscheduled DNA syntheses induced by UV, 4-nitroquinoline 1-oxide, and N-methyl-N'-nitro-N-nitrosoguanidine, were determined. Results confirm that the properties related to XP are not altered by transformation with Epstein-Barr virus and are the same in degrees of defect as are those of dermal fibroblasts from the respective individuals. These XP and normal lymphoblastoid cell lines should be especially useful for biochemical studies on the mechanism of DNA repair, because they are easy to grow in mass culture.


Induction of testicular sarcomas in Fischer rats by intratesticular injection of nickel subsulfide. Nickel subsulfide (Ni3S2) was injected in various amounts into the testis of adult Fischer rats for the study of the acute and chronic effects of Ni3S2 on testicular cells. Rats given injections of 0.6 to 10 mg of Ni3S2 developed an immediate inflammatory response at the site of injection, followed by a delayed, slowly evolving coagulation necrosis of seminiferous tubules and interstitial cells. The extent of testicular necrosis was dose dependent, but at doses of 5 or 10 mg of Ni3S2 the rats invariably developed subtotal destruction of the testis. The testis became atrophic, without regeneration of seminiferous tubules. No damage was seen in the other testis, and no systemic effects were noted. Malignant testicular neoplasms developed in 16 of 19 rats within 20 months after an injection of 10 mg of Ni3S2. These neoplasms were classified by light and electron microscopy as fibrosarcomas, malignant fibrous histiocytomas, and rhabdomyosarcomas. None of the testicular neoplasms was derived from germ cells or genital cord cells. The occurrence of rhabdomyosarcomas in the testis, an organ normally devoid of striated muscle, suggests that Ni3S2 induces malignant transformation of undifferentiated, pluripotential mesenchymal cells.


Decreased alkaline phosphatase in cells transformed by rous sarcoma virus. Chick embryo cells transformed by either of two strains of Rous sarcoma virus (Bryan high titer or Schmidt-Ruppin) have low levels of alkaline phosphatase activity compared with nontransformed chick embryo cells. Essentially no differences in acid phosphatase activity were observed between these transformed and nontransformed cells. A virus mutant, RSV-BH-Ta, induces temperature-dependent transformation in infected cells. At 41 degrees, the transformation-nonpermissive temperature, alkaline phosphatase activities were similar to those of chick embryo cells. Shifting these cells to 37 degrees resulted in a change to transformed morphology and a progressive loss of enzyme activity, requiring 18 to 24 hr to reach the level of transformed cells. Rat embryo cells transformed by murine sarcoma virus also contained lower alkaline phosphatase levels than did nontransformed cells. These observations suggest that decreased alkaline phosphatase activities may be a general property of transformed cells.


In vitro radiation response of cells from four human tumors propagated in immune-suppressed mice. Two recently developed clonogenic assays for human tumor cells have been used to measure the in vitro radiation cell survival of four human tumors, a pancreatic carcinoma, a colonic carcinoma, an oat cell carcinoma of the lung, and a melanoma, propagated as xenografts in immune-suppressed mice. The slopes and shoulders of the survival curves for the first three tumors were all similar with Do's, respectively, of 94, 100, and 131 rads and with Dq's, respectively, of 8, 44, and 41 rads, However, melanoma cells from the fourth tumor had a survival curve that differed from those of the other three, both in having a wider shoulder with a Dq of 216 rads and in having a shallower slope with a Do value of 183 rads. It is suggested that the wide shoulder to the melanoma cell survival curve may in part explain the poor response to small fractionated doses of radiotherapy usually observed clinically for this tumor type. However, the data from the other three tumors suggest that differences in radiotherapeutic response seen in the clinic for these tumors cannot be attributed to differences in intrinsic radiosensitivity of the tumor cells.


Comparative ultrastructural studies of nucleoli of tumor cells treated with adriamycin and the newer anthracyclines, carminomycin and marcellomycin. This study was designed to determine the effects of several antitimor anthracyclines, including Adriamycin and its analogs, carminomycin and marcellomycin, on the ultrastructure of nucleoli of Novikoff hepatoma cells. Adriamycin and carminomycin, which are structurally related, induce nucleolar segregation following the formation of conspicuous fibrillar centers. Marcellomycin did not induce formation of nucleolar fibrillar centers. Instead, numerous microspherules formed following treatment with marcellomycin; later complete nucleolar segregation developed. The microspherules were observed to be in various stages of extrusion from the nucleolar body. This microspherule "migration" appeared to be both time and drug concentration dependent. These results show that the rate and extent of nucleolar ultrastructural aberration may be related to structural differences of the various anthracyclines.


Electron-microscopic observations of normal coelomocytes from the earthworm, Lumbricus terrestris. Coelomocytes of the earthworm, Lumbricus terrestris, were studied by transmission electron microscopy. Four morphological cell types are distinguishable: lymphocytic coelomocytes, granulocytic coelomocytes, eleocytes (chloragogen cells), and inclusion-containing coelomocytes. Within these major categories, several distinct cell types differ and may represent developmental stages. The two types of lymphocytic coelomocytes are small with central nuclei and scanty cytoplasms. Two types of granulocytic coelomocytes differ greatly in shape and content; both have small dark-staining granules that resemble lysosomes. Electrocytes, derived from chloragogen tissue, contain a variety of granules, inclusions and vacuoles. Inclusion-containing coelomocytes appear as two types which may be immature and mature forms. Although these cells resemble those that have been referred to as erythroid cells in other invertebrates, the large inclusion bodies are apparently unrelated to hemoglobin; they can undergo morphologic transformation and be extruded by exocytosis. This information on lymphocytic, granulocytic and inclusion-containing coelomocytes is crucial to understanding more about cellular immunity in the earthworm.


Preferential association of newly synthesized histones with replicating SV40 DNA. The assembly of newly synthesized histones into nucleosomes during replication of SV40 minichromosomes in vivo was studied. Infected cells were labeled with 35S-methionine for a time shorter than that required to complete a round of viral DNA replication. Mature and replicating SV40 minichromosomes were extracted and separated by zonal sedimentation, and their histone content was analyzed by polyacrylamide gel electrophoresis (SDS and acidic urea). We show that the pulse-labeled histones associate preferentially with the replicating DNA.


The binding site on SV40 DNA for a T antigen-related protein. A protein closely related to SV40 T antigen was purified in a biologically active form from cells infected with the defective adenovirus-SV40 hybrid, Ad2+D2. This 107,000 dalton hybrid protein binds and protects a specific portion of SV40 DNA from digestion by pancreatic DNAase I. Hybridization, endonuclease cleavage and pyrimidine tract analysis of the protected fragments reveal that the D2 hybrid protein binds in a sequential manner to tandem recognition sites which lie within a sequence of 120 nucleotides at position 67 near the origin of SV40 replication.


Isolation and characterization of T antigen-negative revertants from a line of transformed rat cells containing one copy of the SV40 genome. Negative selection with FUdR produced revertants from the transformed rat line 14B, which contains one insertion of the SV40 viral genome (Botchan, Topp and Sambrook, 1976). 14B contains nuclear T antigen, grows to a high density, grows in low serum and is anchorage-independent. The revertants fall into three classes with regard to viral DNA sequences: the SV40 DNA is retained; the SV40 DNA is retained but has undergone a deletion; and the SV40 DNA is lost, generating a cured cell. This heterogeneity is not a result of long-term passage. The revertants arise with a frequency of one in 8.4 X 10(5) cells after as few as 12 passages. All three classes of revertants are T antigen-negative, density-sensitive, more serum sensitive than 14B and anchorage-dependent. These data argue for a direct role of the functioning viral genome in the maintenance of the transformed state, and that with 14B, the phenotypes of transformation are not virus gene dosage-dependent.


Implication of developmentally regulated Concanavalin A binding proteins of Dictyostelium in cell adhesion and cyclic AMP regulation. Contact sites A and cyclic AMP phosphodiesterase are Concanavalin A binding membrane proteins. Both are characteristic for the aggregation phase of Dictyostelium discoideum. Extracellular cyclic AMP phosphodiesterase and an inhibitor of this enzyme can be recovered from the extracellular medium by binding to Concanavalin A-Sepharose.


[Histochemical and quantitative methods in the biopsy examination of skeletal muscles]. A complex bioptic examination of skeletal muscles as carried out in 280 excised samples has been subjected to a critical analysis. The examination suggested consists of histological and histochemical processing of native cryostat sections. Besides conventional staining methods the techniques recommended include the reaction for detection of myosin ATP-ase with and without acid preincubation, and reactions for NADH tertazolium reductase, acid phosphatase, nonspecific esterase, glycogen and lipids. Sections stained for ATP-ase are suitable for quantitative examinations such as assessing the diameters of muscle fibres.


Adriamycin stimulated superoxide formation in submitochondrial particles. Adriamycin (doxorubicin), an anticancer agent, stimulated the formation of superoxide in submitochondrial particles isolated from bovine heart. Superoxide formation was detected by oxygen uptake, by the cooxidation of epinephrine to adrenochrome and by the reduction of acetylated cytochrome c. These processes were sensitive to superoxide dismutase (SOD). Rotenone-insensitive oxidation of NADH by the mitochondrial respiratory chain in the presence of oxygen caused the formation of approx 4 nmol of superoxide per min/mg of protein. Adriamycin at a concentration of 400 micron stimulated the rate of superoxide formation 6-fold to 25 nmol.min-1.mg-1, but this was not a maximum rate. Approximately 50 micron adriamycin was estimated to be sufficient for obtaining one-half maximal stimulation. Hydrogen peroxide accumulated as a final reaction product. Measurements of the relative catalase activity of blood-free tissues of rabbits and rats indicated that heart contained 2 to 4% of the catalase activity of liver or kidney. An enhanced production of superoxide and hydrogen peroxide and the relatively low catalase content of heart tissue may be factors in the cardiotoxicity induced by adriamycin chemotherapy if a similar reaction occurs in vivo.


Protein composition of nuclear ribonucleoprotein particles isolated from liver of rats in the early stages of feeding of 3'-methyl-4-dimethylaminoazobenzene and from hepatoma induced by the same carcinogen. The feeding of carcinogenic 3'-methyl-4-dimethylaminoazobenzene (3'-MeDAB) in the early stages results in a change in the protein composition of the nuclear ribonucleoprotein particles of the rat liver. These particles are associated with newly synthesized RNA and it is assumed that they are involved in the processing and in the transport of this RNA. After 6 weeks of feeding of this azocarcinogen, the amount of one of the main polypeptides (apparent molecular weight 42 000) is decreased and after 10 weeks of feeding the particles are devoid of this polypeptide completely. Feeding of the non-carcinogenic p-aminoazobenzene (AB) is without any effect. The loss of this polypeptide is not characteristic for the malignant transformation. In the nuclear ribonucleoprotein particles isolated from hepatoma which has been induced by 3'-MeDAB this polypeptide is present in even higher proportion to other polypeptides than it is in particles isolated from liver cells of control animals. The 3'-MeDAB binds to the proteins of the liver nuclear ribonucleoprotein particles and interferes with the RNA processing. It is proposed that the changes in the composition of the protein moiety of the particles reflect changes in the population of liver cells leading finally to the selection of hepatoma cells which are resistant to the toxic effect of 3'-MeDAB on RNA processing.


Guanosine 3',5'-monophosphate and the action of alkylating agents. The intracellular level of guanosine 3',5'-monophosphate (cGMP) has been measured in Walker carcinoma cells in tissue culture after treatment with various alkylating agents. At concentrations which caused a rise in the level of adenosine 3',5'-monophosphate (cAMP) chlorambucil and 5-(1-aziridinyl)-2,4-dinitrobenzamide (CB 1954) produced only a small (35%) elevation of cGMP, while merophan had no such effect. This suggests that any effect of cAMP will not be outweighed by an equivalent rise in cGMP. Sepcific cytosolic binding of cGMP decreased with increasing resistance of Walker cells to alkylating agents, while the dissociation constant, KD, for binding increased. This was also observed with cAMP binding which suggests that the same protein in responsible for binding both nucleotides.


Persistence of enteroviruses in sewage sludge. Sewage from residential areas often contains viruses pathogenic for man and significant amounts are probably associated with solids in sewage sludge. Information on the survival of viruses in sewage sludge is necessary in order to develop guidelines for recycling programmes that involve spreading the sludge on land. In the present study, a number of enteroviruses were added to sewage sludge and the artificially contaminated sludges were tested for viruses at intervals over a 12-week period. Most of the viruses survived for many weeks at room temperature. It is clear that sewage sludge destined for land application should be adequately treated for virus inactivation. In interpreting these results, it should be borne in mind that the survival of hepatitis A virus might be similar. Recent reports about the reappearance of poliomyelitis in regions with immunization programmes should also be taken into consideration.


Poliomyelitis: epidemiology and prophylaxis. 4. Serological and virological surveys conducted after a mass vaccination campaign for the control of a threatening poliomyelitis epidemic. In 1973, a type 1 poliomyelitis epidemic in Kenya was curtailed at an early stage by two mass distributions of trivalent oral vaccine. It was considered useful to know the immunity status of the child population that had resulted from the vaccine distributions and that had presumably contributed to its control. We also wished to know to what extent wild and vaccine virus strains were in circulation after the mass vaccination campaign. Anal swabs and blood were collected from a sample of the children in four areas where the efficiency of vaccine distribution had varied, and the results of virus isolation attempts and antibody tests are reported. Three poliovirus strains were isolated. It was surprising that, in general, the herd immunity after two vaccination rounds did not substantially differ from that found in Kenya on other occasions. Possible reasons for these results are discussed.


Poliomyelitis: epidemiology and prophylaxis. 5. Results of a two- and three-dose vaccination experiment. In tropical countries, seroconversion rates following oral poliomyelitis vaccination are frequently unsatisfactory. In an area of the Machakos district in Kenya, 4000 children under 5 years of age have been registered and are visited fortnightly by trained field staff as part of a comprehensive, population-based, longitudinal surveillance project. It was considered possible that poliomyelitis vaccine, given orally to children twice or three times at home, would produce satisfactory conversion rates. After collection of blood from a random sample of the children, vaccine was given twice to all children under 5 years of age. Blood was then collected a second time from a different sample of children. After a third dose of vaccine, a third blood collection followed. The percentages of the children that received vaccine each round were calculated and the sera tested for antibodies. It appeared that the mean titre was more strongly related to age than to the number of doses of poliomyelitis vaccine received. Improvement of herd immunity after two vaccine distributions was significant for a few age groups and for two types of vaccine only. In none of the groups was a significant improvement obtained by the third vaccine distribution. The problems associated with vaccination by live poliomyelitis vaccine in tropical countries are discussed in relation to the results.


[Tumor induction in mice treated with hydrocortisone and innoculated with polyoma transformed hamster cells]. Tumors have been induced in hydrocortisone treated Mice innoculated with wild-type polyoma transformed Hamster cells. The filtrate from these tumors infects Mouse embryo cells and induces the production of polyoma virus. The polyoma virus has been characterised in the infected cells with anti-polyoma capsid serum.


A novel alkaline phosphatase, a minor component of normal liver phosphatases. A novel alkaline phosphatase differing from the so-called liver-specific isoenzyme was found in four out of twenty-four normal adult livers. Although the mobility of this enzyme was the same as that of so-called liver-specific alkaline phosphatase on the polyacrylamide gel electrophoretogram, its mobility was not altered following neuraminidase treatment, while that of the liver-specific enzyme was affected by the same treatment. Both enzymes also differed in other enzymatic and immunologic properties. The enzyme, however, resembled the so-called intestinal alkaline phosphatase in many enzymatic and immunologic properties. Thus, the inhibition patterns by amino acids, EDTA and inorganic phosphate, the pH optima, KM values for phenyl phosphate and reactivity with anti-intestinal alkaline phosphatase antibody were quite similar for both enzymes. Differences in the properties of this enzyme and intestinal alkaline phosphatase were in sensitivity to denaturation by treatment with heat and urea and to inhibition by Levamisole. The possible origin of the enzyme in normal liver and its relationship to the Kasahara isoenzyme and fetal intestine-type in hepatoma is discussed.


Effect of clofibrate on the composition of very low and low density lipoprotein subfractions in type III hyperlipoproteinaemia. The effect of clofibrate on the lipid and protein composition of very low and low density lipoprotein subfractions (VLDL of SF greater than 100, 60-100 and 20-60; LDL of Sf 10.4-20, 5.7-12 and 3.5-6.5), was investigated in 6 patients with type III hyperlipoproteinaemia (HLP). After four weeks of therapy significant reductions occurred in the concentration of cholesterol in each VLDL fraction, and of triglycerides in Sf greater than 100 and Sf 60-100 VLDL. No changes were found in the concentrations of apolipoprotein B or of the total tetramethylurea (TMU) soluble proteins, but in four patients in whom polyacrylamide disc gel electrophoresis of the TMU soluble proteins was carried out, it was found that arginine-rich peptide (ARP) had largely disappeared on therapy. These findings would be in keeping with increased catabolism of VLDL in response to clofibrate. No significant changes were observed in LDL lipid or protein concentrations.


Membrane-bound arylamidase from human hepatoma cells having the same electrophoretic mobility as placental membrane-bound arylamidase. A membrane-bound arylamidase from well-differentiated hepatocellular carcinoma having the same electrophoretic mobility as placental membrane-bound arylamidase was found. The enzyme was found to be a sialoprotein and was activated by Co2+. Hepatoma membrane-bound arylamidase had a similar molecular weight (240 000) and kinetic properties to normal liver membrane-bound arylamidase, but differed from the liver enzyme with respect to electrophoretic mobility, heat stability and urea inactivation.


Femoral neuropathy associated with anticoagulant therapy. The English literature concerning femoral neuropathy associated with anticoagulant therapy is reviewed. Anatomical dissections were carried out in order to determine how the neuropathy might be produced. It is postulated that hemorrhage occurs within the iliacus muscle and that a pressure neuropathy of the femoral nerve ensues. Conservative management is recommended. The condition is probably more prevalent than previously thought, as evidenced by the fact that two cases were diagnosed in as many years by one orthopedic surgeon in a 300-bed general hospital.


Epiphyseal cartilage cAMP changes produced by electrical and mechanical perturbations. Epiphyseal cartilage from chick embryo was subjected in vitro for one to 15 minutes to static compressive forces or oscillating electric fields. The same stimuli were applied to suspensions of cells isolated from the epiphyses. At the end of the perturbation cAMP was measured in the tissue or cell extracts by radioimmunoassay. It was found that a physiological (60 g/cm2) static pressure reduced the cAMP content in the tissue and the separated cells. An oscillating electric field above 900 V/1.5 cm, 5Hz, enhanced the cAMP accumulation in the intact tissue. This effect was produced only when the long axis of the bones was oriented parallel to the electric field. In isolated cells an electric field above 750 V/1.5 cm, 5Hz, caused a decrease in cAMP content. Charged matrix macromolecules and orientation of the cells within the cell matrix may have a modulating effect on the initiation of this response. It is postulated that the change in cAMP is the early cellular signal in the response to an electrical or mechanical perturbation which leads to bone remodeling.


Irradiation induced kyphosis.

Irradiation induced kyphosis. Eighty-one patients with Wilms tumor treated by irradiation and chemotherapy were studied. Despite the fact that multiple portals for irradiation were used, each crossing the midline, the amount of irradiation delivered to different parts of the vertebral body varied and it was this variation in delivered dose which produced axial skeletal deformities in 70% of the patients. Of the 57 patients with these deformities 32 had scoliosis, 22 kyphoscoliosis and 3 patients pure kyphosis; 12 patients had a kyphotic deformity of over 25 degrees, 7 patients requiring surgical correction. A high incidence of pseudarthrosis following posterior fusion has led to the preference of a 2-stage procedure, anterior interbody fusion followed by a posterior fusion with Harrington rods after 2 weeks of correction in halo femoral traction.


Effects of renal dopamine receptor and beta-adrenoreceptor blockade on rises in blood angiotensin after haemorrhage, renal ischaemia and frusemide diuresis in the dog. 1. In chloralose-anaesthetized dogs, central venous and arterial angiotensin (AII) levels were monitored by blood-bathed bioassay during venous haemorrhage of 20 ml/kg, acute renal ischaemia induced by suprarenal aortic stenosis and frusemide-induced diuresis. 2. Blockade of intrarenal dopamine receptors with ergometrine reduced markedly the increments in arterial AII associated with haemorrhage or suprarenal aortic stenosis, but did not consistently affect the corresponding increments in venous AII. 3. Ergometrine or renal denervation did not affect the increases of blood AII associated with frusemide diuresis. 4. Blockade of beta-adrenoreceptors with propranolol, by contrast, reduced blood AII increments associated with all three procedures. 5. It is suggested that renin release during moderate haemorrhage and acute suprarenal aortic stenosis is, in the dog, partly due to activation of intrarenal dopaminergic nerves. 6. The possibility is discussed that propranolol may depress renin release in the dog by an action other than that of blocking beta-adrenoreceptors.


Pyrophosphate levels and magnesium oxide therapy in osteogenesis imperfecta. Serum pyrophosphate levels were determined in 52 patients with osteogenesis imperfecta. In 19 patients the levels were increased, in 14 normal and in 19 fluctuated. No correlation with disease severity was evident. By a double blind study it was shown that neither the administration of oral magnesium oxide, nor placebo, affected the serum pyrophosphate level or the clinical course.


Insomnia and the physiology of sleep.

Insomnia and the physiology of sleep. Sleep is a vital human physiologic process. Insomnia can be caused by obsession and depression states, pain, or worry over everyday problems. Because of their pharmacologic action, alcohol and high doses of soporifics used as remedies may produce REM-deficit sleep and actually prolong insomnia. If the true cause of sleeplessness is not recognized and properly treated, insomnia may develop into a severe sleep problem. Since benzodiazepines and chloral hydrate do not suppress REM sleep, they are the medications of choice in the therapy for insomnia.


Biochemical background of atherosclerotic heart lesion in an experiment. The authors investigated enzymic systems and corresponding substrate cycles that transport hydrogen across the mitochondrial membrane in a myocardium with experimental cholesterol-induced atherosclerosis. Sensitive spectrophotometric methods were used for assessing the activities of cytoplasmic and mitochondrial enzymes: lactate and alpha-glycerphosphate dehydrogenases, and of characteristic mitochondrial enzymes: glutamate and beta-hydroxybutyrate dehydrogenases. Specific enzymological methods were used in determining the concentrations of lactic, pyruvic, glycerophosphoric, dihydroacetonephosphoric, malic, oxaloacetic, glutamic, alpha-ketoglutaric, acetoacetic, and beta-hydroxybutyric acids. The cytoplasmic NAD/NADH quotient was calculated. The investigators found a reduction of enzymic activities in the "shuttle" transport system studied, marked deviations from normal levels of their substrates, and a reduction of the NAD/NADH quotient by the factor 0.56. All these phenomena represent a biochemical background of a complex of symptoms characterizing severe myocardial lesion in experimental atherosclerosis.


Serum lipids in angiographically assessed coronary atherosclerosis. Cholesterol, triglycerides, and Lp(a)/pre-beta1 lipoprotein were analyzed in 153 patients typed for liproprotien patterns. Coronary atherosclerosis was determined by selective coronary angiography and graded by a system taking into account proximal, middle and distal segments. Smoking habits, family history and hypertension were also recorded. Normal coronary arteries were encountered in 45, moderate coronary atherosclerosis (less than median score) in 50, and severe changes (greater than median score) in 58 patients. Cholesterol (P less than 0.05), positivity of Lp (a)/pre-beta1 lipoprotein (P less than 0.01), a family history of coronary heart disease (P less than 0.05), and smoking (P less than 0.01) differed between the group of normal arteries and the whole group of luminal obstructions. Serum triglycerides were not associated with coronary atherosclerosis. Cholesterol, positivity of the Lp(a)/pre-beta1 lipoprotein and a family history of coronary heart disease were also associated with the severity of the disease. Smoking was less prevalent in the group with severe changes.


Patterns of cyclic 3'5'-adenosine monophosphate excretion in parathyroid dysfunction. The pattern of excretion of cyclic 3'5' adenosine monophosphate (cAMP) over 24 h was evaluated in nine hyperparathyroid, four hypoparathyroid and three postoperative hyperparathyroid patients. The hyperparathyroid patients had increased excretion at the time of the normal nadir and were normal or low occasionally during the day. The pattern in the hypoparathyroid and in the postoperative patients also differed from normal. These findings suggest that both peaks of urinary cAMP excretion found in normal individuals result from the effects of parathyroid hormone on the kidneys.


Iron, zinc, free radicals and oxygen in tissue disorders and cancer control. Evidence is presented supporting the argument that various tissue disorders are the result of iron becoming decompartmentalized. In healthy cells, vital molecules are protected from the action of decompartmentalized iron by the presence of zinc. This protection is particularly important during processes leading to cell division. Should excessive decompartmentalization occur and this protective mechanism become overloaded or should it be weakened by ill health, damaging oxidative free-radical reactions may take place. If these are extensive, death may result. When they are only limited, possibly owing to a low oxygen tension or the presence of copper or a carcinogen, the cell may survive but cancer may result. This 'antioxidant' theory of cancer may provide a unifying mechanism for the action of many carcinogenic agents. Carcinogens are considered to be activated not only by the more usually accepted enzymic pathways but by free-radical reactions, catalysed by iron, in the vicinity of critical sites.


Environmental effects on the central nervous system. The central nervous system (CNS) is designed to respond to the environment and is peculiarly vulnerable to many of the influences found in the environment. Utilizing an anatomical classification (cortex, cerebellum, peripheral nerves) major toxins and stresses are reviewed with selections from recent references. Selective vulnerability of certain areas to particular toxins is apparent at all levels of the CNS, although the amount of damage produced by any noxious agent depends on the age and genetic substrate of the subject. It is apparent that the effects of certain well known and long respected environmental toxins such as lead, mercury, etc., deserve continued surveillance. In addition, the overwhelming impact on the CNS of social damages such as trauma, alcohol, and tobacco cannot be ignored by environmentalists. The effect of the hospital and therapeutic environment has become apparent in view of increased awareness of iatrogenic disorders. The need for particular laboratory tests, for example, examination of CSF and nerve conduction toxicity studies, is suggested. Epidemics such as the recent solvent neuropathies suggest a need for continued animal studies that are chronic, as well as acute evaluations when predicting the potential toxic effects of industrial compounds.


Effect of isometric exercise on catecholamines in the coronary circulation. Arterial and coronary sinus blood levels of catecholamines, adenosine 3', 5'-cyclic monophosphate (c-AMP) and lactate were measured during isometric exercise in fourteen patients. In no patient did lactate production occur. Mean resting total catecholamine levels both arterial (0.53 +/- 0.07 ng/ml; 2.94 +/- 0.38 nmol/l) and coronary sinus (0.4 +/- 0.08 ng/ml; 2.22 +/- 0.44 nmol/l), did not change significantly on exercise. Coronary sinus c-AMP levels fell on exercise from 11.5 +/- 0.8 nmol/l (resting) to 9.9 +/- 0.8 nmol/l (exercise) (P less than 0.01) with an arterial-coronary sinus difference of 1.2 nmol/l (P less than 0.01) on exercise. Our findings suggest that isometric exercise does not normally result in excessive cardiac symphathetic activity.


The role of iron-sulfur center 2 in electron transport and energy conservation in the NADH-ubiquinone segment of the respiratory chain in Paracoccus denitrificans. 1. The electron paramagnetic resonance spectra at 15 K of reduced membrane particles of Paracoccus denitrificans exhibit resonance signals with g values, line shapes and temperature profile which are similar to the signals of the iron-sulfur centers observed in the NADH-ubiquinone segment of mitochondrial respiratory chains. These iron-sulfur centers are reducible with NADH, NADPH as well as chemically with dithionite. 2. Sulphate-limited growth of Paracoccus denitrificans results in the loss of an electron paramagnetic resonance signal (gz approximately 2.05, gy approximately gx approximately 1.92) which has properties similar to those of iron-sulfur center 2 of the NADH dehydrogenase of mitochondrial origin. The loss of this signal is accompanied by a decrease in the NADH oxidase and NADH ferricyanide oxidoreductase activities to respectively 30 and 40% of the values found for succinate-limited growth conditions. In addition respiration in membrane particles from sulphate-limited cells loses its sensitivity to rotenone. 3. Since sulphate-limited growth of Paracoccus denitrificans induces loss of site I phosphorylation [Arch. Microbiol. (1977) 112, 25-34] these observations suggest a close correlation between site I phosphorylation, rotenone-sensitivity and the presence of an electron paramagnetic resonance signal with gz approximately 2.05 and gy approximately gx approximately 1.92.


Changes in enzyme activities during the artificially stimulated transition from follicular to luteal cell types in rat ovary. This paper reports the time course of development of the intramitochondrial cholesterol side-chain-cleavage activity, cytoplasmic NADP+-dependent isocitric dehydrogenase, malic enzyme and glucose-6-phosphate dehydrogenase during ovarian maturation, using as a model the immature rat ovary stimulated to develop with pharmacological doses of gonadotrophin. These enzymic activities were correlated with increases in ovarian content of DNA, cellular content of adenosin 3':5'-monophosphate, and the levels of plasma progesterone. The plasma progesterone concentrations followed closely the development of the [4-14C]cholesterol side-chain-cleavage which was mimicked by the cytoplasmic isocitric dehydrogenase; both enzymes increased in activity 28 times during the 6 days of this study. There was no correlation between adenosine 3':5'-monophosphate levels and cholesterol side-chain cleavage or progesterone plasma concentrations.


Metal-free and metal-substituted cytochromes c. Use in characterization of the cytochrome c binding site. The luminescent properties of metal-free, tin(IV) and zinc(II) cytochromes c have been used to characterize the interaction of cytochrome c with mitochondria and cytochrome oxidase. Diminution in the fluorescence yields of tin and zinc cytochrome c occur when these derivates bind to cytochrome oxidase or mitochondria. Based upon spectral overlap and quantum yield, the distance between the porphyrin rings of cytochrome a and cytochrome c is estimated according to Forster theory to be in the neighborhood of 3.5 nm. Measurements of the polarized emission of metal-free 'porphyrin' cytochrome c when bound to oriented layers of cytochrome c oxidase indicate that the porphyrin is bound obliquely to the plane of the oxidase layers with an angle of about 70 degrees C from heme plane to membrane plane. It is proposed that these data have significance for elucidation of electron transfer mechanisms.


Intermediates in the conversion of procollagen to collagen. Evidence for stepwise limited proteolysis of the COOH-terminal peptide extensions. Intermediates in the conversion of procollagen to collagen were isolated from radioactively labeled chick cranial bones by ion-exchange chromatography. Cleavage of these proteins with vertebrate collagenase revealed that each of the several forms of these intermediates lacked NH2-terminal but retained COOH-terminal extensions. The chain composition of each intermediate was resolved by two-dimensional slab gel electrophoresis. The intermediates differed from each other in having sustained cleavages in zero, one or two pcalpha chains. The relative proportions of intermediates with different intact pcalpha chains, observed in conversion of procollagen, have enabled us to construct a detailed model of the stepwise limited proteolysis of procollagen.


Fate of histone messenger RNA in mengovirus-infected Ehrlich ascites tumor cells. Histone mRNA was isolated from mengovirus-infected Ehrlich ascites tumor cells at various times postinfection and quantitated in a reticulocyte cell-free protein-synthesizing system. The amount of translatable histone mRNA decreases during the first hour postinfection by 30%, rises during the following 1-1.5 h by 10-15%, drops progressively in the further course of infection, and reaches 20% of the control at the end of the infectious cycle (8-9 h postinfection). On the basis of the relative histone mRNA contents, the histone-synthesizing potentials of mengovirus-infected Ehrlich ascites tumor cells are substantially higher throughout infection than actually expressed in vivo. This result indicates that the virus-induced shutoff of histone synthesis is not directly the consequence of inactivation or degradation of histone mRNA. Most of the histone mRNA recovered from mengovirus-infected Ehrlich ascites tumor cells is bound to ribosomes. Late in infection, certain mRNAs are co-isolated with histone mRNAs, very likely due to loss or shortening of poly(A) occurring after release of the mRNAs from polyribosomes.


The crystal structure of complexes between horse liver alcohol dehydrogenase and the coenzyme analogues 3-iodopyridine-adenine dinucleotide and pyridine-adenine dinucleotide. We have studied the binding of the enzymatically active NAD+ analogue, 3-iodopyridine-adenine dinucleotide, and the inactive analogue, pyridine-adenine dinucleotide to the enzyme horse liver alcohol dehydrogenase using X-ray crystallographic methods. These studies were made under such conditions that crystals of the complexes were isomorphous to apoenzyme crystals. Both analogues bind in the same conformation. The binding of the adenosine moiety is very similar to that of ADP-ribose or NADH bound to the enzyme. The conformation and mode of binding of the remaining portions of the analogue molecules is, however, quite different. The pyridine ring is not situated in the active-site pocket as the nicotinamide group in the isomorphous enzyme-NADH-imidazole complex but lies at the surface of the crevice between the two domains of the subunit, approximately 1.5 nm away from the catalytically active zinc atom. Lys-228 which has been shown to be important for NADH dissociation is in this region of the molecule.


Kinetic studies on the transport of cytoplasmically synthesized proteins into the mitochondria in intact cells of Neurospora crassa. The transport of cytoplasmically synthesized mitochondrial proteins was investigated in whole cells of Neurospora crassa, using dual labelling and immunological techniques. In pulse and pulse-chase labelling experiments the mitochondrial proteins accumulate label. The appearance of label in mitochondrial protein shows a lag relative to total cellular protein, ribosomal, microsomal and cytosolic proteins. The delayed appearance of label was also found in immunoprecipitated mitochondrial matrix proteins, mitochondrial ribosomal proteins, mitochondrial carboxyatractyloside-binding protein and cytochrome c. Individual mitochondrial proteins exhibit different labelling kinetics. Cycloheximide inhibition of translation does not prevent import of proteins into the mitochondria. Mitochondrial matrix proteins labelled in pulse and pulse-chase experiments can first be detected in the cytosol fraction and subsequently in the mitochondria. The cytosol matrix proteins and those in the mitochondria show a precursor-product type relationship. The results suggest that newly synthesized mitochondrial proteins exist in an extra-mitochondrial pool from which they are imported into the mitochondria.


Quantitative determination of myoinositol, inositol 1-phosphate, inositol cyclic 1 : 2-phosphate and glycerylphosphoinositol in normal and Rous-sarcoma-virus-transformed quail fibroblasts under different growth conditions. Myoinositol and its phosphorylated derivatives have been quantitatively determined in normal and Rous-sarcoma-virus-transformed quail cells under various growth conditions using [2-(3)H]myoinositol at isotope equilibrium conditions. The following amounts were determined (nmol/mumol phospholipid, as a unit of cell mass): exponentially growing normal and tumor cells contained 25--40 nmol free inositol, 0.40--0.45 nmol myoinositol 1-phosphate, 0.30--0.50 nmol glycerylphosphoinositol, and 0.03--0.04 nmol myoinositol cyclic 1 : 2-phosphate. At high cell populations in the absence of serum, conditions which result in cessation of growth by normal but not by tumor cells, changed levels were found for glycerylphosphoinositol and free inositol. In tumor cells the levels of these two compounds increased to 0.64 nmol and 64 nmol, respectively. In normal cells glycerylphosphoinositol increased to 0.95 nmol and free inositol showed highly elevated levels of 144 nmol. At short pulses the specific activities of inositol 1-phosphate and inositol cyclic 1 : 2-phosphate were found to be higher than that of phosphatidylinositol. This was not the case for glycerylphosphoinositol.


The binding of calcium and magnesium to sarcoplasmic reticulum vesicles as studied by manganese electron paramagnetic resonance. The binding pattern of the biologically relevant ions calcium and magnesium has been investigated via the binding  of the ion-analogue manganese. The binding parameters of manganese are obtained conveniently by standard electron paramagnetic resonance techniques, the binding of calcium and magnesium is inferred from competition experiments. The quantitative analysis was carried out with a computer program which was able to treat the competition of three kinds of ions for three classes of independent and one class of cooperative binding sites. It was possible to correlate specific binding classes with biological functions of the sarcoplasmic reticulum membrane. The magnesium and manganese specific binding class of medium affinity is related to the catalytic function of these ions in the ATP-splitting. For the first time a manganese and calcium specific class of cooperative binding sites has been observed and it can be related to the inhibition of the calcium transport at high concentrations of manganese or calcium. From the binding results a model can be developed which allows a full description of the role of the divalent ions and their specific binding sites during calcium transport.


The structure of human-plasma low-density lipoprotein B. An X-ray small-angle scattering study. 1. X-ray small-angle scattering of human plasma lipoprotein B of the low-density fraction (rho = 1.016--1.060 g.cm-3) has been recorded to high precision at different electron density contrasts. 2. The overall structure of the particles is characterized by a quasi-spherical shape and radial symmetry. A maximum diameter of 23 nm and a molecular weight of 2.4 X 10(6) have been determined. 3. The internal structure is described in terms of a model consisting of spherical layers with different electron densities indicating that the neutral lipids are arranged in the core of the molecule up to a radius of about 8 nm surrounded by a monolayer of free cholesterol, phospholipids and protein. The neural lipids are shown to be in an ordered, liquid crystalline state at 4 degrees C and to undergo a thermotropic transition into a disordered state at higher temperatures.


Effect of methylxanthines on binding of the glucocorticoid receptor to DNA-cellulose and nuclei. The binding of [3H]dexamethasone-receptor complex from rat liver cytosol to isolated nuclei or DNA-cellulose can be greatly enhanced at low temperature by the presence of theophylline. Aminophylline and caffeine can mimic this effect; however, papaverine and 1-methyl-3-isobutylxanthine, at concentrations inhibitory to phosphodiesterase, are without effect on glucocorticoid receptor binding to DNA. Furthermore, theophylline can be added when adenosine 3':5'-monophosphate-(cAMP) hydrolysis is already complete and still enhance DNA binding. These results imply that this effect of theophylline is independent of its known effect on cAMP levels. Activation by methylxanthines does not alter the sedimentation of the glucocorticoid-receptor complex in sucrose gradients but does alter the pI and in this respect brings about changes resembling those which occur upon activation by heat. Recently we have shown that pyridoxal phosphate inhibits the binding of heat-activated receptor to DNA-cellulose. Similarly, we have shown here that pyridoxal phosphate also inhibits the DNA-cellulose binding of theophylline-treated receptor. The presence of theophylline also enhances the rate of binding of [3H]dexamethasone to the receptor and increases its apparent affininty for the steroid. The data suggest that the effect of theophylline is on some cytosolic component, perhaps the receptor itself. Enhanced DNA binding as a result of exposure to theophylline at low temperature can also be demonstrated using the glucocorticoid receptor of kidney, thymus and Reuber H35 cells.


Two mechanisms of isoproterenol inhibition of smooth muscle. The relaxation of depolarized ideal and uterine smooth muscle by isoproterenol was compared with that brought about by dibutyryl-cAMP and by other agents which increase cytoplasmic cAMP. Qualitative differences observed between the time courses of the relaxations imply the existence of a rapid beta-adrenergic mechanism independent of cAMP as well as a slower cAMP-dependent relaxation. This dual process model for beta-adrenergic inhibition of smooth muscle is consistent with seemingly opposing published reports.


Spinal branching of rubrospinal axons in the cat. The branching patterns of rubrospinal (RS) axons projecting to the cervical spinal cord between C3 and C8 were studied in the cat. RS neurons were identified by their antidromic responses to microstimulation of local axon branches within the cervical gray matter. Twenty-six of 58 RS neurons projecting to the cervical gray matter also sent axon branches to the thoracic spinal cord. Two out of 40 of these RS neurons also sent axon branches to the lumbar spinal cord. Using a collision technique, it was demonstrated that stem axons of rubrospinal neurons commonly sent multiple collaterals to different cervical segments. Neurons projecting to the cervical spinal cord alone were located in the dorsal quadrants of the red nucleus. Those projecting to cervical, as well as to more caudal segments, were intermingled with the former, and in slightly more ventral portions of the red nucleus. The presence of RS neurons projecting to widely separate levels of the spinal cord suggests that individual RS neurons may be capable of ultimately influencing two or more different motoneuron pools.


Properties of the synaptic transmission of the newly formed cortico-rubral synapses after lesion of the nucleus interpositus of the cerebellum. 1. Properties of synaptic transmission during and after repetitive activation of the newly formed cortico-rubral were examined in the red nucleus neurons (RN) of cats after lesions of the nucleus interpositus of the cerebellum (chronic cats) as well as in normal ones. 2. A prominent facilitation of the amplitude of cortico-rubral unitary EPSPs was observed in both normal and chronic cats when a stimulus to the cerebral peduncle (CP) was preceded by another stimulus by 2-50 msec. 3. Time course of the facilitation shows that it attains maximum at the interval of about 3 msec and decays approximately exponentially lasting for 50 msec or more. 4. When three successive stimuli of identical intensity were applied to CP, the degree of facilitation was more prominent than that for double shock. 5. There was a positive correlation between the time to peak of the cortico-rubral EPSPs and their maximum value of facilitation. 6. The posttetanic potentiation of the cortico-rubral EPSPs was observed after tetanic stimulation to CP in chronic and normal cats. It lasts for a few minutes in both cases.


Ultrastructural responses of the hypoglossal nucleus to the presence in the tongue of botulinum toxin, a quantitative study. The ultrastructural effects of local injection of botulinum toxin into the left half of the tongue of the rat, were studied quantitatively 35 days postoperatively in the left hypoglossal nucleus. The results showed (1) a decrease in somatic and neuropil bouton numbers because of loss of boutons with symmetrical synapses and clear spherical synaptic vesicles, (2) a decrease in the numbers of dendrite profiles in the neuropil, (3) an increase in the proportion of dendrites and boutons with unusual inclusions, suggestive of profile retraction, (4) an increase in the proportion of profiles which were unusually electron-dense, (5) an increase in the amount of astrocyte, and a growth of astrocyte sheaths around bouton-free neurone surfaces, (6) the presence of occasional microglia, and (7) subastrocytic subsurface cisterns. Control rats injected with boiled toxin had no responses except (3) and (4) above, and then only to a modest extent, possibly due to mechanical damage of a few axons or terminals at the time of injection, or to insufficient inactivation of the toxin by boiling. The results were compared with those at 35 days after axotomy, and it was concluded that botulinum toxin, which interrupts neuromuscular transmission, elicits the same responses in the hypoglossal neurones, as does transection of the hypoglossal nerve, even though earlier studies had discovered no glial replication after botulinum toxin, in contrast to axotomy.


Study of histochemical and biochemical changes in the liver of rats induced by high doses of isoprenaline. The aim of the present work was to study by means of histochemical and chemical methods the 7-day course of changes in carbohydrate metabolism in the liver of male rats induced by a single dose of isoprenaline of 50 mg/kg administered subcutaneously. A statistically significant reduction was seen both in the level of free glycogen and lactate within 24 hours. The decrease of pyruvate level was not so marked. At the same time, there was increased, and within the hepatic lobules also extended activity of enzymes catalyzing glycogenolysis, i.e. alpha-glucan phosphorylase and particularly the branching Q-enzyme, glucose-6-phosphatase and LDH, whereas the level of malate and activity of SDH, which are constituents of the Krebs cycle, were found to be reduced. Cytochrome oxidase activity was changed after 24 hr compared to the controls. The obtained results indicate that an extensive glycogenolysis occurs in the liver of rats in the 24 hr following s.c. administration of isoprenaline, the major part of liver glycogen being degraded through glucose-6-phosphate to blood glucose and its metabolism via the Krebs cycle reduced. The observed metabolic changes are of reversible character and tend to normalize over the 2nd and 3rd day following isoprenaline administration.


Serum activity inhibiting specific simian virus 40-induced transplantation resistance and its correlation with primary SV40 tumors appearance in hamsters. Using the modified technique of transplantation test, ITR serum activity was found in most (14 out of 21) individual hamster sera obtained during the latent period of primary SV40 carcinogenesis (60 days after virus infection when newborn). On the other hand, as a rule, no ITR activity was observed in the sera of the same hamsters after tumor appearance and during their growth. ITR activity rapidly disappeared from sera of hamsters neonatally infected with SV40 after their successful immunization with the same virus during the latent period. There appears to be a correlation between the presence of ITR serum factor during the latent period and the subsequent primary SV40 tumor appearance in hamsters.


Investigations into the effects of ACTH on the half-life of mitochondrial proteins in the rat adrenal cortex. ACTH elongates the half-life of the mitochondrial proteins from the rat adrenal cortex, and chloramphenicol inhibits this effect of ACTH. The hypothesis is advanced that the ACTH-provoked stabilization of the adrenocortical mitochondrial proteins requires continuous mitochondrial DNA-dependent protein synthesis.


Modulation of transmission in different electronic junctions by aminopyridine. 4 distinct electronic inputs can be identified in amphibian motoneurons perfused with calcium-free solution containing manganese. The differential effect of 4-aminopyridine on different electronic junctions may reflect the peculiarities of molecular architecture of potassium channels in different electrically excitable presynaptic membranes.


GABAergic inhibition of neurons in the ventral tegmental area. Stimulation of the nucleus accumbens evokes a potent inhibition in neurons of the ventral tegmental area. GABA is likely to act as a transmitter in this descending inhibitory system.


[Temporal characteristics of sleep in the dog]. Main time characteristics of sleep were studied in 9 adult dogs in experiments with 24-hr polygraphic recordings. The all-day sleep quantity (35%), circadian distribution of sleep with the maximum between 24 and 9 hours, more rapid rise of delta-sleep (DS) in the second half of day and of REM sleep in the first hours of night -- all these data suggest that the dog sleep schedule is under considerable influence of ecological factors related to time organization of activity and sleep in humans. A relatively high amount of drowsiness in dog's sleep (nearly 25%) is comparable with DS representation. REM occupies about 20% of total sleep time. Mean REM cycle lasts 30 min. The data obtained can be used as the standards of dog sleep in neurophysiological, pharmacological and behavioral studies.


[Local temperature characteristics of the isolated cortex in wakefulness and alteration of the sleep stages]. In cats, changes of the brain electric activity were followed by local temperature shifts in isolated and in intact cortex: the transition from slow wave sleep to paradoxical sleep and from quiet to active wakefulness were accompanied by increasing of the cortex temperature, while transition from paradoxical sleep to slow sleep decreased the temperature. During falling asleep and alternation of the sleep stages, both in isolated and intact cortex, the temperature shifts preceded the electric activity changes. The temperature of isolated cortex is by 0.2 degrees lower than that of the opposite intact hemisphere.


[Effect of anticholinesterase agents on the protein and RNA content in the cells of the supraoptic nucleus during sleep and its disorder]. In rats, administration of anticholinesterase drugs (both the penetrating blood-brain barrier armin and the nonpenetrating preparation Gd-42) prevented the decrease of protein content in the neurons and glia of the supraoptic nucleus, as well as of RNA content in the neurons which usually occurred in intact animals after 24-hr deprivation of the REM sleep by the Jouvet method (also with partial disturbance of the slow wave phase).


[Effect of ACTH and hydrocortisone on the functional state and interaction of brain structures]. In chronic experiments on cats ACTH facilitated the development of behavioral responses to gradually increasing electric stimulation of the medial hypothalamus, hippocampus and the midbrain reticular formation as well as potentiated the reticulo-hypothalamic influence. The effect of hydrocortisone was opposite. The reciprocal reticular and hormonal influences on the basal and anterior amygdala were revealed as well as some hormonal changes due to unspecific thalamic effects upon the limbic structures. The revealed systemic hormonal effect on the state and interaction of cerebral structures seems to be the base for integration of neural and hormonal mechanisms of some behavioral responses.


[Regulation of hormone receptor (author's transl)]. The regulation of hormone receptors is exerted through genetic and environmental factors. Genetic control of hormone receptors is analyzed by studying changes in their characteristics during the ontogeny, phylogeny, and malignant transformation of cells. The pattern of changes in glucocorticoid receptor contents in cytosol fraction during ontogeny is quite different from tissue to tissue. In the rat's lung tissue, glucocorticoid receptor contents are high during the fetal period and decrease promptly after birth, whereas the contents in the rat's liver are low during the fetal life and start to increase after birth. Glucocorticoid receptors in thymocytes of rats during T-cell differentiation increase transiently, which gives a high sensitivity to steroid treatment in the immature thymocytes. The induction of glucocorticoid receptors in different tissues at different stages of life is strictly controlled by the genetic program. Whether the induction is mediated by humoral factor(s) remains to be clarified. Amounts of glucocorticoid receptors in a particular tissue are subjected to species differences. In a group of steroid-sensitive animals such as the rat, mouse, rabbit and hamster, the amount of glucocorticoid receptors in liver sytosol is rich, whereas it is poor or undetectable in a group of steroid-resistant animals, such as the human, monkey, dog and guinea-pig. Changes in hormone receptors are also observed in malignant tumors. Under certain circumstances, hormone receptors "appear" in malignant tumors originated from organs in which no detectable hormone receptor is observed. These tumors may be called hormone receptor-producing tumors. Environmental factors are also important in regulating hormone receptors. Down regulation of hormone receptors is observed in insulin and glucocorticoids.


[Urinary excretion of 3beta-hydroxy-delta5-steroids in the newborn infant and the effect of ACTH administration (author's transl)]. The urinary 3beta-hydroxy-delta5-steroids excreted during the period of the first week by newborn infants were analyzed by means of gas chromatography and gas chromatography-mass spectrometry to study the changes in the amount of these steroids. The changes in the amount of each steroid after the administration of ACTH were also studied. The results are summarized as follows: 1) Seven kinds of 3beta-hydroxy-delta5-steroids were identified in the urine of the newborn infants. 17alpha, 21-dihydroxypregnenolone was newly identified. 2) The daily changes in the amount of each of these steroids differed. Generally, the amount of the C-19 steroids increased day by day and that of the C-21 steroids decreased during the period of our observation. 3) After the administration of ACTH, the amount of these urinary steroids excreted increased at nearly the same rate.


Cell-mediated cytotoxicity: comparison of primary and secondary immune reactions after parainfluenza type 1 virus inoculation. H-2 antigen compatibility of effector spleen cells and target cells was necessary for cell-mediated cytotoxicity (CMC) after both 1 and 2 intraperitoneal inoculation of 6/94 virus into mice. T cells were responsible for both primary and secondary CMS reactions. Neither reaction was influenced by the presence of virus-specific antibody added to the test system. Secondary CMC peaked earlier than the primary response and declined more rapidly. In vitro stimulation of primed spleen cells could also be used to detect the secondary CMC response.


Cell-mediated immunity to vesicular stomatitis virus infections in mice. The T cell-mediated immune responses of mice against vesicular stomatitis virus (VSV) were assessed by measuring direct primary foot pad swelling after local VSV infection and cytotoxic activity in spleens. The cytolytic activity was mediated by T cells since it was anti-theta + complement sensitive, was restricted by the K and D region but not the I region of H-2 and rapidly increased after 4 days but decreased 8 days after systemic or local infection. Cytolytic activity was virus-specific as reciprocally tested with VSV and vaccina virus immune T cells. Measurable activity on day 7 depended on infectious virus dose, virus virulence, and non-H-2 genetic background of the host. More than half of the cytolytic activity wasblocked specifically by either immune anti-H2 or rabbit anti-VSV antisera. Analysis of the kinetics of appearance of antigenic changes using metabolic inhibitors, revealed that the changes that rendered target cells susceptible to lysis after infection, occurred within the first hour after infection.


Gangliosides of active and inactive neuroblastoma clones. It is possible to divide neuroblastoma cells into clones able to synthesize neurotransmitters (active clones) or not (inactive clones). The analysis of gangliosides of active and inactive clones shows that their total lipid sialic acids is markedly lower than that of neuron-enriched fractions prepared from brain. The ganglioside pattern of the cultured cells also differs notably from those obtained with neuronal fractions from brain. The absence of tri- and tetrasialogangliosides and the presence of appreciable amounts of the simplest monosialogangliosides are particularly noticeable in the neuroblastoma. Morphological differentiation obtained by serum deprivation, dibutyryl cyclic AMP or bromodeoxyuridine does not restore a true neuronal pattern. Gangliosides could not therefore be used as a marker of neuronal differentiation in this type of cell. No correlations can be found between the ganglioside pattern and the ability of cells to synthesize neurotransmitters.


An attempt to differentiate selection and amplification in hormone receptor development: the unicellular model. Earlier experiments demonstrated that a membrane pattern of Protozoa behaves as a receptor with respect to hormones of higher organisms. This raised the possibility that some selection or strengthening of this unspecific patter is involved in the evolution of the specific membrane patterns of the individual cells of higher organisms. In the present experiments, as a result of continual histamine treatment, the phagocytotic ability of a population of Tetrahymena was increased more intensely than after a single histamine treatment. The phagocytotic rate remained high for some time after the animals were returned from histamine-containing to normal medium. Thus, from the experimental data, it appears likely that as the hormone appears, selection becomes involved in the proliferation of cells possessing receptors. This observation might not only be of phylogenetic interest, but could also be relevant in receptor maturation as manifested in ontogenetic membrane differentiation.


Immunosupressive activity of the basic protein from tumor tissue. The basic protein prepared from AH-130 ascites hepatoma cells was tested for its effect on the cell-mediated immune response of Donryu strain rats. The basic protein suppressed the response of peripheral and lymph node lymphocytes to phytohemagglutinin and lymphocyte trapping in the antigen-draining lymph node. These results indicated that the basic protein from tumor tissue possibly had an immunosuppressive activity and affected inadequately the cell-mediated immune response as usually observed in tumor-bearing hosts.


Hematological alterations in tumor-bearing rats, with reference to pathogenesis of chronic type of disseminated intravascular coagulation syndrome. Hematological studied were carried out serially in the rats transplanted subcutaneously with Yoshida ascites hepatoma AH-109A. Significant changes were observed in fibrinogen level, fibrinogen degradation products, recalcification time, platelet count, and fragmentation of red blood cells. Formation of thrombi was revealed in the vessels of tumor tissue morphologically from early stage when the tumor grew to a palpable size. Thrombi were formed also in the arterioles of the lungs in the terminal stage. Bleeding tendency was noted in some cases at death. These findings suggested the experimental induction of a type of disseminated intravascular coagulation. The systemic changes of the blood occurring in the terminal stage were preceded by localized intravascular coagulation and fibrinolysis in the tumor in early stage of tumor growth.


Effect of 1-(2-chloroethyl)-3-(beta-D-glucopyranosyl)-1-nitrosourea on experimental tumors. A newly synthesized water-soluble nitrosourea derivative, 1-(2-chloroethyl)-3-(beta-D-glucopyranosyl)-1-nitrosourea (GANU; NSC-254157) has marked activities against experimental tumors such as lymphoid leukemia L-1210, ascites sarcoma- 180, ascites hepatoma AH-130, and Walker carcinosarcoma-256. A single intraperitoneal injection of the compound is more effective than successive injections on the L-1210 system. An especially interesting point is that both the single intraperitoneal and single oral administrations 2 days after tumor implantation showed a high effectiveness. In addition, growth of the mammary adenocarcinoma transplanted subcutaneously on the back of mice was also considerably suppressed by successive intraperitoneal injections of the compound.


Evaluation on the recurrent cases of early gastric carcinoma. In the First Department of Surgery, Faculty of Medicine, Chiba University, 213 cases with early gastric carcinoma have undergone an excision from 1955 to 1975. The observed 5-year, relative 5-year, observed 10-year, and relative 10-year survival rates of these cases amount to 89.3%, 99.8% and 98.0% respectively. The present investigation concerns 13 of these patients with diagnosed postoperative recurrent cases. The post-operative gastric carcinima recurrence might be classified to metastasis to liver and lymphnodes, peritoneal dissemination, residual gastric recurrence and recurrence in miscellaneous organs. The 13 cases involved 2 cases of liver metastasis, 9 cases of residual gastric recurrence and 2 cases of recurrence in miscellaneous organs. Nine of these cases underwent re-operations which were grouped to 6 cases of residual gastrectomy, 2 cases of excision of miscellaneous organs (cecum and rectum), and 1 case exploratory laparotomy. The development in the cases is characterized by 3 cases of stump persistent carcinoma at the residual stomach due to stump carcinoma positive in excision, 2 cases of residual gastric carcinoma due to lymphatic vessel invasion and 6 cases of multiple carcinoma.


Methadone blockade of 2-deoxyglucose-induced pancreatic secretion in the rat. The effects of methadone on pancreatic exocrine secretions in the rat were tested under basal conditions and after hormonal stimulation by secretin and caerulein or after stimulation of the differentially acting cholinergic agents acetylcholine, 2-deoxyglucose, and electrical stimulation of the vagus. Methadone had no effect on basal hydroelectrolytic secretion. It decreased basal enzyme secretion very slightly under our experimental conditions. The stimulatory effects of 75 mg of 2-deoxyglucose per kg were completely blocked by methadone at 5 mg per kg and this blockade was reversed by nalorphine at 6 to 9 mg per kg. It was found that there are doses of methadone (100 microgram) which block 2-deoxyglucose effects when injected into brain ventricles but are ineffective when systemically introduced. The effects of secretin, caerulein, acetylcholine, and electrical stimulation of the vagus were not depressed by methadone. These results strongly suggest that the methadone blockade of 2-deoxyglucose effects occurs at a central level and is mediated by narcotic drug receptors.


The histoenzymic activity of gyrus cinguli in the course of postnatal ontogeny of the rat. Histochemical investigations concerning the activity of several dehydrogenases and hydrolases in the area cinguli during postnatal ontogenic development of the rat were performed. Brain sections corresponding to the region of area cinguli, obtained from rats aged 1, 3, 8, 17, 40 and 60 days postnatal, were subjected to histochemical assays for various dehydrogenases, phosphatases and esterases. The developmental changes concerning histoenzymic reactivity of the callosal gyrus with regard to several respiratory and hydrolytic enzymes were assessed and described. Considerable differences in enzymic reactivity, appertaining particularly to the phosphatases and esterases, between the anterior and posterior parts of the callosal gyrus were found. The unusual enzymic reactivity of glial cells in the individual regions of the area cinguli has been pointed out.


The activity of some intracellular oxidative enzymes in the thyroid follicular epithelium of Xenopus laevis Daud. in experimental long-term hypokinesia. Histoenzymological methods were applied to examine the activities of previously little studied intracellular oxidative enzymes in the cells of the thyroid follicular epithelium of Xenopus laevis Daud. specimens kept in aquarium. Succinate, lactate, alpha-glycerophosphate, glucose-6-phosphate and reduced NAD and NADP dehydrogenases and cytochrome oxidase were studied. In comparison with other chordate species a very strong activity was revealed particularly by lactate dehydrogenase. The data obtained suggest that the metabolism in thyroid cells of the motionless Xenopus is based on glycolysis mainly.


[Contractile responses of the guinea pig ileal longitudinal muscle to antispasmodics (author's transl)]. Papaverine, benactyzine and Aspaminol were found to induce strong contractions of guinea pig ileal longitudinal muscle in a medium containing 3.2 mM of Ca2+. The contractile actions of papaverine, benactyzine and Aspaminol were potentiated with increase in Ca2+. The contractile actions of papaverine, benactyzine and action of papaverine was completely blocked by Mn2+, which by itself caused a contraction of the longitudinal muscle. The maximal contraction by papaverine was obtained at the concentration of more than 10(-5) M, WHILE BOTH BENACTYZINE AND Aspaminol induced the maximal contractions at the concentration of 3 X 10(-4) M. During and after the contraction by papaverine, contractile responses to benactyzine and Aspaminol were not observed. Changes in the intracellular level of cyclic GMP after exposure of the longitudinal muscle to papaverine, benactyzine or Aspaminol did not occur. These results suggest that papaverine, benactyzine Aspaminol may induce contractions of the ileal longitudinal muscle through a similar mechanism of action. A likely assumption is that these drugs move intracellular bound Ca2+ to the contractile protein of the muscle.


Non-functioning islet cell tumours of the pancreas: a review of radiological literature and a report on two cases. Two cases of endocrine inactive malignant islet cell tumors diagnosed by angiography are presented. The angiographic features are described and the differential diagnosis of the tumor and its metastases are discussed. Radiological literature of the past decade on this subject is reviewed.


[The treatment of severe and recurrent nonspecific colpitis]. Attention is drawn to the incidence of colpitis and the symptom of "vaginal discharge" in the gynaecological field, as well as to the required investigations as far as differential diagnosis is concerned. Severe, purulent, unspecific colpitis and also recurrent colpitis require particularly careful treatment. Excellent therapeutic results have been achieved over many years with repeated local administration of TC-Gel-Steraject and, if necessary, additional treatment with an oestriol preparation; this therapy is therefore recommended.