Cellular cyclic AMP in dispersed mucosal cells from guinea pig stomach. In dispersed mucosal cells from guinea pig stomach cyclic AMP was increased 4-fold by theophylline, 5-fold by prostaglandin E2, and 10- to 15-fold by histamine. Theophylline augmented the increase in cellular cyclic AMP caused by histamine or prostaglandin E1 and the actions of histamine and prostaglandin E1 were additive. Cellular cyclic AMP was not altered by carbachol, gastrin, secretin, vasoactive intestinal peptide, glucagon, insulin or the octapeptide of cholecystokinin. Metiamide or diphenhydramine but not atropine inhibited the increase in cellular cyclic AMP caused by histamine, but did not alter the concentration of cyclic AMP in control cells or in cells incubated with theophylline or prostaglandin E1.


Characterization of the oligosaccharide side chain of apolipoprotein C-III from human plasma very low density lipoproteins. Apolipoprotein C-III1 and apolipoprotein C-III2 each contain one oligosaccharide side chain, bound O-glycosidically to threonine in position 74 of the amino acid sequence. The studies reported in this paper characterize these alkali labile oligosaccharides, thereby demonstrating the complete structure of apolipoprotein C-III. Monosaccharide analysis revealed the following sugar composition: D-galactose/N-acetyl-D-galactosamine/sialic acid 1 : 1 : 1 and 1 : 1 : 2 for apolipoprotein C-III1 and apolipoprotein C-III2, respectively. Treatment of desialylated apolipoproteins with alkaline borohydride released the reduced disaccharide beta-D-galactosyl-(1 leads to 3)-N-acetyl-D-galactosaminitol, which was detected by gas-liquid chromatography. Further studies employing periodate oxidation and Smith degradation indicated that the structure of the trisaccharide from apolipoprotein C-III1 was alpha-N-acetylneuraminyl-(2 leads to 3)-beta-D-galactosyl-(1 leads to 3)-N-acetyl-D-galactosaminitol. The tetrasaccharide structure from apolipoprotein C-III2 is made up of this trisaccharide plus one sialic acid residue linked to C6 of N-acetyl-D-galactosaminitol, as was shown by the assessment of chromogens formed upon alkaline degradation.


Studies on the utilization of ferritin iron in the ferrochelatase reaction of isolated rat liver mitochondria. The utilization of ferritin as a source of iron for the ferrochelatase reaction has been studied in isolated rat liver mitochondria. 1. It was found that isolated rat liver mitochondria utilized ferritin as a source of iron for the ferrochelatase reaction in the presence of succinate plus FMN (or FAD). 2. Under optimal experimental conditions, i.e., approx. 50 micromol/1 FMN, 37 degrees C, pH 7.4 and 0.5 mmol/l Fe(III) (as ferritin iron), the release process, as shown by the formation of deuteroheme, amounted to approx. 0.5 nmol iron/min per mg protein. 3. The release process could not be elicited by ultrasonically treated mitochondria, lysosomes, microsomes or cytosol, i.e., the release of iron from ferritin was due to mitochondria and was a function of the in situ orientation of the mitochondrial inner membrane. 4. The release of iron from ferritin by the mitochrondria might be of relevance not only for the in situ synthesis of heme in the hepatocyte, but also with respect to the mechanism(s) by means of which iron is mobilized for transport to the erythroid tissue.


Properties of beta-adrenergic receptors in untreated and butyrate-treated Hela cells. HeLa cells contain receptors on their surface which are beta-adrenergic in nature. The binding of (-)-[3H]dihydroalprenolol is rapid, reversible, stereospecific and of relatively high affinity. The HeLa cells also contain an adenylate cyclase which is activated by (-)-isoproterenol greater than (-)-epinephrine greater than (-)-norepinephrine. The adenylate cyclase of HeLa is also activated by guanyl-5'-ylimidodophosphate (Gpp(NH)p), a nonhydrolyzable analogue of GTP. Inclusion of both (-)-isoproterenol and Gpp(NH)p leads to approximately additive rather than synergistic activation of adenylate cyclase. After treatment of HeLa cells with 5mM sodium butyrate there is an increase in the number of beta-adrenergic receptors, but not in their affinity, which is reflected in an increased ability of (-)-isoproterenol to activate adenylate cyclase. Other properties of the beta-adrenergic receptor including association and dissociation rates, temperature optimum of adenylate cyclase and response to Gpp(NH)p are relatively unaffected by butyrate pretreatment of the cells.


Solubilization of pituitary receptors for thyrotropin-releasing hormone. Receptors for thyrotropin-releasing hormone were solubilized by Triton X-100. Membrane fractions from GH3 pituitary tumor cells were incubated with thyrotropin-releasing hormone in order to saturate specific receptor sites before the addition of detergent. The amount of protein-bound hormone solubilized by Triton X-100 was proportional to the fractional saturation of specific membrane receptors. Increasing detergent:protein ratios from 0.5 to 20 led to a progressive loss of hormone . receptor complex from membrane fractions with a concomitant increase in soluble protein-bound hormone. The soluble hormone . receptor complex was not retained by 0.22 micron filters and remained soluble after ultracentrifugation. Following incubation with high (2.5--10%) concentrations of Triton X-100 and other non-ionic detergents, or following repeated detergent extraction, at least 18% of specifically bound thyrotropin-releasing hormone remained associated with particulate material. Unlike the hormone receptor complex, the free hormone receptor was inactivated by Triton X-100. A 50% loss of binding activity was obtained with 0.01% Triton X-100, corresponding to a detergent:protein ratio of 0.033. The hormone . receptor complex was included in Sepharose 6B and exhibited an apparent Stoke radius of 46 A in buffers containing Triton X-100. The complex aggregated in detergent-free buffers. Soluble hormone receptors were separated from excess detergent and thyrotropin-releasing hormone by chromatography on DEAE-cellulose. Thyrotropin-releasing hormone dissociated from soluble receptors with a half-time of 120 min at 0 degrees C, while the membrane hormone . receptor complex was stable for up to 5 at 0 degrees C.


Follitropin receptors in rat testis. Characterization with enzymatically 125I-labeled human follitropin. The interaction between enzymatically radioiodinated human follitropin and the follitropin receptors in testis homogenate was investigated in immature and adult rats. The 125I-labeled human follitropin exhibited high binding activity with specific binding of up to 17% in the presence of an excess of testis homogenate. Approx. 50% of the bound hormone could be eluted at pH 5, and the receptor purified tracer exhibited a 3.6-fold increase in binding activity when compared with the original tracer preparation. Quantitative analysis of equilibrium binding data was performed with corrections for the measured specific activity and maximum binding activity of the tracer hormone. The equilibrium association constants (Ka) determined 24 degrees C were not significantly different in immature and adult rat testis, and the mean value for Ka was 3.9 . 10(9) M-1. At 37 degrees C, the Ka value obtained using immature rat testis was 1.3 . 10(10) M-1. The association of 125I-labeled human follitropin with immature rat testis homogenate was time and temperature dependent. In the presence of an excess of unlabeled hormone, 30--60% of the preformed hormone . receptor complex was dissociated after 24 h incubation. A specific and sensitive radioligand-receptor assay for follitropin was developed using immature rat testis homogenate. The minimum detectable dose of purified human follitropin was 0.6 ng, and human urinary and pituitary follitropin, ovine follitropin and pregnant mare serum gonadotropin reacted in the assay with equivalent slopes. The potencies of highly purified pregnent mare serum gonadotropin and highly purified human  follitropin were similar in the radioligand-receptor assay, consistent with the follitropin bioactivity of the equine gonadotropin.


Catabolite repression of isocitrate lyase in methylamine-grown Pseudomonas MA. Effect of carbon and nitrogen sources. The synthesis of the C1-type isocitrate lyase found during growth of Pseudomonas MA on methylamine was investigated. It was shown that this enzyme is subject to catabolite repression by preferred carbon sources, e.g., succinate, and by ammonia. The carbon repression can be overcome by cyclic AMP, which was shown to be acting at the transcriptional level. Repression by ammonia is overcome during growth with methylamine as sole nitrogen, but not carbon, source. Uptake experiments showed that the uptake of methylamine from the medium was prevented by ammonia in the presence, but not in the absence, of an alternative carbon source. Measurement of cyclic AMP levels in cells grown on methylamine and on succinate, glycerol, glucose and acetate as carbon sources (with ammonium chloride as nitrogen source) revealed that methylamine-grown cells have the lowest cyclic AMP level despite having the highest C1-type isocitrate lyase activity. Cells grown on acetate with methylamine as sole nitrogen source possess both C1-type and C2-type isocitrate lyase. The results indicate that the synthesis of C1-isocitrate lyase is under control by repression-derepression involving a specific inducer, cyclic AMP, and an effector whose action is related to the nitrogen supply of the cell.


Hormone-specific responses and biosynthesis of sulfolipids in cell lines derived from mammalian kidney. The established cell lines isolated from mammalian kidney were characterized by its receptor activities against hormones and the ability to synthesize sulfolipids localized in the renal tubule. The level of 3':5'-cyclic AMP in JTC-12.P3 (monkey kidney) cells increased in 2 min as much as 2.5-5-fold on activation with 1.0 unit/ml of bovine parathyroid hormone or 1.9 units/ml of synthetic parathyroid hormone (1-34) resulting in intracellular cyclic AMP concentration of more than 40 pmol/mg protein. Prostaglandin E1 (14 micronM) and isopropylnorepinephrine (10 micronM) were also found to increase the concentration of cyclic AMP by more than 30- and 9-fold, respectively. Addition in medium of calcitonin, arginine vasopressin, adrenocorticotropic hormone and glucagon caused no significant changes of cyclic AMP level in the cell. In contrast, MDCK, a cell line isolated from canine kidney, reacted to arginine vasopressin, isopropylnorepinephrine and prostaglandin E1 and only slightly to parathyroid hormone. MDBK cell line derived from bovine kidney or fibroblast cell lines from rat lung and guinea pig kidney did not react to any of the hormones specific to kidney, i.e. arginine vasopressin, calcitonin or parathyroid hormone in the presence of theophylline. However, in the presence of 2 mM isobutylmethylxanthine, small but significant elevation of cellular cyclic AMP levels in response to calcitonin, arginine vasopressin, isopropylnorepinephrine and prostaglandin E1 was observed. The cell lines JTC-12, MDCK and MDBK, when incubated with H235SO4, incorporated the isotope into sulfolipids assigned as sulfatides and ceramide dihexoside sulfate or in MDCK also into cholesterol sulfate. The results suggested that JTC-12, MDCK and MDBK cell lines are epithelial origin and also JTC-12 and MDCK originated most probably from renal tubular cells of cortex and medulla, respectively.


Adrenocorticotropin-induced unresponsiveness in cultured adrenal tumor cells. Our results demonstrate that adrenocorticotropin (ACTH)-induced refractoriness occurs in cultured adrenal tumor cells. Cells became 85% refractory to ACTH-induced cyclic AMP formation in 20 min and the effect persisted if the hormone remained in the incubation medium. Refractory cells gradually regained hormone-specific responsiveness within 24 h if cultures were incubated in fresh media containing serum. The observed effect is hormone specific since cyclic AMP could not induce unresponsiveness to ACTH. The addition of ACTH plus inhibitors of protein synthesis partially reversed hormone-specific refractoriness. However, preincubation with cycloheximide or diphtheria toxin led to superinduction of ACTH-induced cyclic AMP formation. These experiments suggest that unresponsiveness, following hormonal activation of adrenal cells, may be related to a decrease in hormone-specific binding sites or to synthesis of an adenylate cyclase inhibitor.


Identification of the regulatory steps in gluconeogenesis in cotyledons of Cucurbita pepo. 1. The aim of this work was to discover the steps at which the conversion of oxaloacetate to glucose 6-phosphate during gluconeogenesis is regulated in the cotyledons of 5-day-old seedlings of Cucurbita pepo. 2. We estimated the maximum catalytic activities of all the enzymes in the above sequence and also the amounts of their substrates present in vivo. The results show that the reactions catalysed by fructose-1,6-bisphosphatase and phosphoenolpyruvate carboxykinase are the only ones in the sequence that are substantially displaced from equilibrium in vivo. 3. We also determined the effects of 3-mercaptopicolinic acid, an inhibitor of gluconeogenesis, on the amounts of the gluconeogenic intermediates present in vivo. The results show that the enzyme system, fructose-1,6-bisphosphatase: phosphofructokinase, and the system phosphoenolpyruvate carboxykinase: phosphoenolpyruvate carboxylase make major contributions to the regulation of gluconeogenesis in the cotyledons. 4. Possible mechanisms for the above regulation are discussed.


Azidophenantridinium compounds as photoaffinity labels of cholinergic proteins. The synthesis of diazidopropidium and diazidoethidium is described. The applicability of these compounds as photoaffinity labels for cholinergic proteins has been investigated: diazidopropidium inhibits neuromuscular transmission. This inhibition is reversible if the compound is applied in the dark but becomes irreversible after irradiation with white light. Inhibition is accompanied by a disappearance of miniature endplate potentials. Electrophysiological analysis of this effect indicates that diazidopropidium acts postsynaptically by blocking the acetylcholine receptors. At the molecular level the action of diazidopropidium and diazidoethidium on acetylcholinesterase has been investigated: both compounds appear to bind to a peripheral acetylcholine binding site of this enzyme. Binding of 125I-labeled alpha-neurotoxin from Naja naja siamensis to purified membranes from Torpedo californica electric tissue rich in acetylcholine receptors is diminished after incubation and irradiation with diazidopropidium. About half of the toxin binding sites appear to be blocked by the photoaffinity label.


Acidic-phosphoprotein phosphatase activity of rat ventral prostate nuclei: apparent lack of effect of androgens. A protein phosphatase activity has been demonstrated in nuclei of rat ventral prostate utilizing 32P-labelled phosvitin as a model acidic phosphoprotein substrate. This phosphoprotein phosphatase has a pH optimum of 6.7, is unaffected by the sulphydryl protecting agent 2-mercaptoethanol, and requires a divalent cation for maximal activity. Of the various divalent cations tested, Mg2+ is the most effective in reactivating the EDTA-inhibited enzyme. The phosphatase is inhibited by sodium flouride, sodium oxalate, N-ethylmaleimide, ATP and ADP but is relatively insensitive to ammonium molybdate. Increased ionic strength of the reaction medium also causes a reduction in the enzyme activity, e.g., by 48% at 200 mM sodium chloride. The activity of the acidic phosphoprotein phosphatase did not change significantly at 48 h or 96 h post-orchiectomy when expressed per unit of nuclear protein. However, it is reduced by approx. 30% at these times after castration if based on DNA content. The decline in activity per nucleus reflects the decrease in the realtive nuclear protein content observed at 48 h or 96 h post-orchiectomy. This suggests that the decline in the phosphorylation of prostatic nuclear acidic proteins which occurs upon androgen withdrawal is not due to increased nuclear phosphatase activity.


Comparison of large unilamellar vesicles prepared by a petroleum ether vaporization method with multilamellar vesicles: ESR, diffusion and entrapment analyses. Large unilamellar vesicles, prepared by a petroleum ether vaporization method, were compared to multilamellar vesicles with respect to a number of physical and functional properties. Rotational correlation time approximations, derived from ESR spectra of both hydrophilic (3-doxyl cholestane) and hydrophobic (3-doxyl androstanol) steroid spin probes, indicated similar molecular packing of lipids in bilayers of multilamellar and large unilamellar liposomes. Light scattering measurements demonstrated a reduction in apparent absorbance of large unilamellar vesicles, suggesting loss of multilamellar structure which was confirmed by electron microscopy. Furthermore, large unilamellar vesicles exhibited enhanced passive diffusion rates of small solutes, releasing a greater percentage of their contents within 90 min than multilamellar vesicles, and reflecting the less restricted diffusion of a unilamellar system. The volume trapping capacity of large unilamellar vesicles far exceeded that of multilamellar liposomes, except in the presence of a trapped protein, soy bean trypsin inhibitor, which reduced the volume of the aqueous compartments of large unilamellar vesicles. Finally, measurement of vesicle diameters from electron micrographs of large unilamellar vesicles showed a vesicle size distribution predominantly in the range of 0.1--0.4 micron with a mean diameter of 0.21 micron.


Superoxide dismutase of the eye: relative functions of superoxide dismutase and catalase in protecting the ocular lens from oxidative damage. 1. Activities of superoxide dismutase (superoxide: superoxide oxidoreductase, EC 1.15.1.1) have been estimated in eye tissues. In rabbit eye, superoxide dismutase is present in corneal epithelium, corneal endothelium, lens, iris, ciliary body and retina. In lens the activity is in capsule epithelium. 2. Copper chelator diethyldithiocarbamate inhibited lens superoxide dismutase in vitro and in vivo in rabbit. 3. H2O2 caused inhibition of superoxide dismutase activity of lens extract, and this inhibition was potentiated by the catalase inhibitor 3-amino-1H-1,2,4-triazole (3-aminotriazole) or NaN3. 3-Aminotriazole or NaN3 had no effect on lens superoxide dismutase. Thus endogenous catalase of lens affords protection to the lens superoxide dismutase from inactivation by H2O2. 4. In rabbit having early cataract (vacuolar stage) induced by feeding-3-aminotriazole, there was a decrease in superoxide dismutase of lens, a fall in ascorbic acid of ocular humors and lens, and a 2--3-Fold increase in H2O2 of aqueous humor and vitreous humor. We conclude that catalase of eye affords protection to the lens from H2O2 and it also protects superoxide dismutase of lens from inactivation by H2O2. Superoxide dismutase, in turn, protects the lens from the superoxide radical, O2.-. It is likely that inhibition of these enzymes may lead to production of the highly reactive oxidant, the hydroxyl radical, under pathological conditions when H2O2 concentration in vivo exceeds physiological limits as in cataract induced by 3-aminotriazole. A scheme of reaction mechanism has been proposed to explain the relative functions of ocular catalase and superoxide dismutase. Such a mechanism may be involved in cataractogenic process in the human.


alpha-Hydroxy-beta-keto-gamma-aminobutyric acid in human urine. A new amino acid has been isolated from the normal human urine. The chemical structure of the amino acid was determined to be alpha-hydroxy-beta-keto-gamma-aminobutyric acid based on its physical properties involving NMR, infrared and mass spectra, as well as chemical degradation and synthesis. In six healthy adults the urinary contents of the new amino acid were 3.2--4.5 mumol/24 h.


The role of calcium and cyclic adenosine 3',5'-monophosphate in the regulation of glycogen metabolism in phagocytozing human polymorphonuclear leukocytes. Incubation of human polymorphonuclear leukocytes in a glucose-free Krebs-Ringer bicarbonate buffer for 2 h resulted in glycogen depletion, decreased phosphorylase activity and increased synthase-R activity. Addition of dialyzed latex particles to starved leukocytes revealed a very rapid ingestion rate (half-maximal ingestion within 30 s). This uptake is followed by glycogenolysis associated with an immediate two-fold increase in phosphorylase a activity and a synthase-R to -D conversion within 30 s. Furthermore, in rapid time-course experiments with phagocytozing cells we found that the concentration of cyclic AMP increased by 93% within 15 s and returned to baseline values at 1 min. In a medium without added calcium and with 1 mM ethyleneglycol-bis-(beta-aminoethylether)-N,N'-tetraacetic acid, phagocytosis was blocked, cyclic AMP formation decreased by 50% and phosphorylase activation was abolished, but the conversion of synthase-R to -D was preserved. Addition of calcium ions to cells suspended in a calcium-free buffer without added latex results in phosphorylase activation and glycogenolysis, but not in cyclic AMP increase or synthase-R to -D conversion. Measurements of 45Ca efflux during phagocytosis suggest an initial increase in cytosolic calcium obtained by a release of membrane-bound 45Ca. Activation of phosphorylase during phagocytosis is thus presumably due to an increase in cytosol Ca2+ and subsequent activation of phosphorylase kinase, and is independent of the simultaneous increase in concentration of cyclic AMP. Phosphorylation of synthase R to the D form does not depend on the presence of Ca2+ in the extracellular phase.


[Low temperature reduction of cytochrome c complexes]. The absorption spectra of ferricytochrome c complexes with azide, imidazole and cyanide reduced by trapped electrons at the liquid nitrogen temperature were investigated. Differences have been found between the absorption spectra at T=77 degrees K of cytochrome c complexes reduced by radiation and the absorption spectrum of ferrocytochrome c. These differences are supposed to be due to the axial ligation of the Fe(II) by exogenous ligand. The temperature increase led to the removal of exogenous ligand from Fe(II).


[Spin-label progesterone binding to serum albumin]. The binding of spin label progesterone to bovine serum albumin was studied by the spin-probe technique. The binding capacity of protein was established. It was shown that protein formed a rigid complex with steroid, the correlation time of this complex being 50 ns. In the complex the radical part of the steroidal molecule has a hydrophobic environment.


[Effect of ferricyanide, dark adaptation and aging on the properties of the EPR I signal of chloroplasts]. Ferricyanide (10-3 M) is shown to oxidize from 25 to 50% of the total P700 pool in the dark. The ferri-ferrocyanide mixture thus obtained, acting as redox buffer, accelerates the P700+ dark reduction after continuous far red illumination is turned off or after a saturating flash. The oxidized P700 yield on the flash increase in the presence of ferricyanide as a result oxidation of species in the neighborhood of P700 centers. During aging (24 h at 2--3 degrees in the dark) the yield of flash-induced P700 oxidation also increases thus indicating the loss of cyclic electron flow in aged chloroplasts.


[Limiting behavior of a model of the impulse activity of a neuron for large frequency of input flow and small contributions of a single synapse (non-diffusion approximation)]. The limit behavior of probability models of neuron's firing, viz. models with exponential decay and constant threshold is considered. These models differ from each other by F(v), distribution of jumps of potential after arrival of input impulse. If average number of input impulses arriving during the decaying time of one impulse in very large and threshold depolarisation, measured by unit of average jump of potential, is large too, than limit density of interspike interval distribution does not depend on form of F(v); and depends on other neuron's parameters mean and variance of F(v). Moreover, the form of this density depends on only one parameter--normalised threshold. Under this condition interspike interval distribution for diffusion model converges to the same limit. Lapase transform for limit distribution is found and results of its numeric inverts are given.


Relationships in hydrogen metabolism between hydrogenase and nitrogenase in phototrophic bacteria. Purple bacteria Rhodospirillum rubrum and Thiocapsa roseopersicina form two enzymes, hydrogenase and nitrogenase, which participate in hydrogen metabolism. H2 photoproduction in these bacteria is associated mainly or completely with the action of nitrogenase. The soluble and membrane-bound hydrogenases of T. roseopersicina have similar physicochemical properties (mol. weight, subunit composition, N-terminal amino acids, Fe2+ and S2- content, pl. Eo'). In comparison with other hydrogenases the enzyme from R. rubrum and T. roseopersicina evolve H2 with high rate from reduced cytochrome c3, but not from ferredoxins. H2 production and N2 fixation take place in the presence of NAD(P)H. NADP-reductase, ferredoxin and cytochrome c3 participate in this reaction. Possible relationships between hydrogenase-nitrogenase in the metabolism of molecular hydrogen are discussed.


EPR determination of the oxidation-reduction potentials of the hemes in cytochrome c3 from Desulfovibrio vulgaris. EPR spectroscopy in conjunction with oxidation-reduction potentiometry has been used to determine the half-reduction potentials of the four hemes of cytochrome c3. As predicted, the four hemes of cytochrome c3 have different mid-point potentials. The Em values are: Heme I,--284 mV; Heme II,--310 mV; Heme III,--324 mV and Heme IV,--319 mV. The n-values in each case was near one.


[Spike transmission in statistical neuronal ensembles. IVa. Stating the problem in a diffusion approximation]. An area of tissue in the field CA3 of the Hippocampus has been selected controled by a basket cell. The functioning of the basket cell has been described by a point approximation method forthe transport equation, and that of the ensemble of pyramidal cells by a diffusion approximation one. Furthermore, the area of tissue is broken up into N2 of "cells" wherein a transition is effected from the diffusion equation of the N2 of the point equations involved. If the sizes of "cells" are commeasurable to the diffusional length of the pyramidal cell collaterals each "cells" being relatively independent generator of activity. In the latter case the fundamental interaction type is the mutual negative influence of the "cells" through the basket cell. Should the area be broken up into further, the mutual influence of the "cells" becomes noticeable and cannot be neglected. The behaviour of spatially homogenous diffusion approximation model is equivalent of the point approximation model.


NMR studies of electron carrier proteins from sulphate reducing bacteria. The sulphate-reducing bacteria have a complex electron transfer system which leads to the reduction of sulphate by oxidation of either organic substrates or molecular hydrogen. These bacteria can either produce or consume molecular hydrogen. The central part of this electron pathway for Desulovibrio gigas is constituted by hydrogenase (3 X (4Fe-4S)). cytochrome c3 (4 haems with different redox potentials) and a one (4Fe-4S) cluster ferredoxin. This ferredoxin is isolated in different oligomeric forms, which stabilize different oxidation states and have different physiological roles; the trimer FdI being involved in the production of H2 and the tetramer FdII being more efficient for the consumption of H2. The presence of intrinsic probes (the iron ions) in these proteins is particularly helpful for structural studies using NMR spectroscopy. These studies allowed a characterization of the oxidation states used by the different oligomers of the ferredoxin and obtaintion of structural information on multi-haem cytochromes (c3 and c7). NMR is also suitable to study protein-protein interaction. The study of the complex formed between FdII and cytochrome c3 has shown that there is an alteration of the kinetics of electron transfer upon complexation.


[Anion-sensitive adenosine triphosphatase from membranes of rat red blood cells]. Anion-sensitive ATPase was solubilized from membranes to rat red blood cells. The effect of bicarhonate, sulfite and perchlorate on the activity of ATPase was studied. Close resemblance of the properties of ATPase of rat red blood cells and of mitochondrial ATPase was observed.


[Study of the reaction between steroids and serum albumin by the spin probe method]. The data obtained from the study of interaction of two steroids with serum albumin shows that D-ring of steroid molecule has more hidrophobic environment then A-ring in the complexes of steroid with protein. The plane of steroid is perpendicular to the long axis of protein molecule.


[Allosteric regulation of ceruloplasmin activity]. Study of the interactions of homogenous human ceruloplasmin preparations with histamine show that the rate of p-phenylene diamine oxidation by ceruloplasmin is increased in the presence of histamine; the increase in the enzyme activity is independent of histamine concentration. The dependence of the reaction rate on substrate concentration is S-shaped, both in the presence and in the absence of histamine. The respective values of the Hill coefficient and Rs for the enzyme in the presence and in the absence of histamine are 2.5 and 2.0 and 8.0 and 10.4. Histamine does not change ceruloplasmin-specific absorption at 610 nm. Evidence from EPR studies show that histamine does not interact with Cu of the enzyme active center. During interaction with histamine the antigenic properties of the enzyme are changed. Histamine increases the oxidase activity of the enzyme in human and rat blood sera and exerts multifold effects on the enzyme activity in patients with hepatolenticular degeneration. After injection of histamine to rats the enzyme activity is increased without a simultaneous increase in Cu concentration in the blood serum, i.e. without de novo synthesis of ceruloplasmin. The data obtained suggest that ceruloplasmin is probably an allosteric enzyme, which histamine is its positive allosteric effector.


A new animal model for schizophrenia: interactions with adrenergic mechanisms. Amphetamine-induced stereotyped behavior in animals is proposed as a model for schizophrenia. Chronic amphetamine administration produces stereotyped behavior and a paranoid schizophreniform syndrome in man, whereas in animals a behavioral sensitization to stereotypy is evoked. We now show that phenylethylamine (PEA), an amphetamine-like stimulant concentrated in the limbic system of human brain, produces stereotypy in rats with a behavioral sensitization when chronically administered. In comparing amphetamine-induced stereotypy with PEA-induced stereotypy, we found that the alpha-adrenergic blocking agents phentolamine and phenoxybenzamine selectively antagonize PEA stereotypy, whereas the beta-adrenergic blocking agent propranolol fails to alter significantly stereotypies evoked by PEA or amphetamine administration. Catecholamine depletion by alpha-methyl-p-tyrosine administration blocks stereotypies induced by both PEA amphetamine, whereas selective norepinephrine depletion antagonizes only PEA stereotypy; the amino acid precursors of both norepinephrine and dopamine potentiate stereotypies. Therefore, PEA-elicited stereotypy, but not amphetamine-elicited stereotypy, is dependent upon norepinephrine; the significance of this for the PEA animal model of schizophrenia is discussed.


Spatially rearranged vision and REM sleep: a lack of effect. An observation first made in our laboratory was responsible for focusing our attention upon an apparent increase in REM sleep in relation to altered visual experiences during the day. This finding was consistent with certain information-processing theories of REM sleep function and seemed to offer experimental support for them. Therefore, we proceeded to conduct a series of closely related experiments, designed to investigate further the possible REM-augmenting effects of spatially rearranged vision. Although our first two experiments seemed to support the original observation, subsequent, more carefully designed experiments, did not. The combined results from all of our experiments forced us to conclude that there is no consistent effect of distorted daytime vision on the amount or percentage of REM sleep. REM latency, or the number of rapid eye movements during REM periods. We suggest that future research on the topic employ experiments specifically designed to explore relationships between personality variables and a subject's REM sleep response to altered sensory input.


Renal transplantation and viral infections. III. Clinical and virological correlations. Studies of the serological development after 26 renal transplants confirm the high frequency of antibody rises not only against the herpes virus group, but also against other virus groups such as measles, Coxsackie B viruses. These antibody rises correlate with febrile episodes and hepatic dysfunction in which CMV is the most often involved. However, the frequency of antibody rises against various viral antigens without any clinical event to suggest viral etiology; the lack of concomitant virus isolation (except the herpes group), as well as the ocurrence of simultaneous antibody rises against several viruses, all suggest that some of these various antibody rises observed may be related to immunological dysfunction rather than to virus infection.


Abnormal lipoproteins in a case of primary biliary cirrhosis. The serum of a patient diagnosed as a primary biliary cirrhosis was studied during the various evolutive stages of the disease. During the non-icteric period, the serum lipoprotein had the chemical composition (rich in cholesterol and phospholipids) of that found in cholestasis; however the bilirubinemia was normal, and no LPX was detected; there was a normal esterified to total cholesterol ratio and the alphaLP had not decreased, as expected, but increased and was divided into two fractions; immunoelectrophoretic studies of ultracentrifugal fractions showed some LP with HDL immunological properties, but with lower densities than that of normal HDL. We suggest that the phospholipid overloading of some of these HDL in the patient's serum could explain the changes in density. Some months after this study, the patient's serum exhibited the characteristics found in cholestasis, with disappearance of HDL and appearance of LPX. During the subsequent period, the serum again contained phospholipid rich HDL and LPX. In fact several LPX with different densities were found. It seems, therefore, that in cholestasis, LP undergo modification in which the HDL peptide chains play an important role.


Alteration of cellular ribonucleases associated with murine oncogenic virus infection. The ribonuclease activity of peripheral lymphocytes from Balb/c mice  was studied at various intervals subsequent to infection of mice by oncornavirus. Lymphocytes from mice infected with Friend leukemia virus possessed elevation of RNase activity within 8 days subsequent to infection. Balb/c mice infected with Moloney sarcoma virus demonstrated an analogous elevation of RNase activity with 7-9 days postinfection. Diminishment of cellular RNase activity occurred in the Friend leukemia model concomitant to the occurrence of significant numbers of erythroblasts in the peripheral blood, while ribonuclease activity in lymphocytes from mice infected with Molney sarcoma virus returned to normal 1-2 weeks subsequent to host rejection of tumor. It is concluded that elevation of RNase activity within the lymphocyte represents an early event in oncogenic viral infection within these two tumor models. The possible meaning of elevation of RNase activity is a target (the lymphocyte) not predestined to undergo neoplastic transformation is discussed.


Experimental infection of Callithrix Jacchus marmosets with Herpesvirus ateles, Herpesvirus saimiri, and Epstein Barr virus. We inoculated common marmosets (Callithrix Jacchus) with Herpesvirus ateles (HVA), Herpesvirus saimiri (HVS), and Epstein-Barr virus (EBV). HVA-induced tumors contained several cell types, including giant cells reminiscent of the Sternberg-Reed cells observed in human Hodgkin's disease. HVS and EBV did not induce tumors, although HVS was present in lymphocytes and elicited a strong antibody response. EBV elicited only a variable antibody response. We feel that more common marmosets should be used to determine if the pathologic and immunologic lesions caused by HVA would be a suitable animal model for Hodgkin's disease and/or other malignant lymphomas of man. Inconsistency in the induction of tumors by EBV and HVS in common marmosets suggets that this species may be a different type of model for human cancer research than the cottontop marmoset, which is the most susceptible animal host for EBV and HVS oncogenesis.


Microtubules, cyclic AMP and lysosomal enzyme release. A review. The importance of microtubules and cyclic AMP in the control of lysosomal enzyme release from polymorphonuclear leukocytes is questioned. The article is a short review of pertinent literature designed to stimulate further research into this important mechanism of inflammation.


The Epstein-Barr virus (EBV) in human pathology. II. Serologic profiles of EBV infections. Antibodies to EBV induced intracellular antigens, (VCA, EA, NA) and VCA-IgM antibodies have been investigated to define EBV serologic profile of 245 individuals. This profile was a first studied in EBV primary infections. In infectious mononucleosis, the efficiency of EBV serodiagnosis is lower than Paul Bunnel Davidsohn reaction (PBD) : primary infection profile is only characterised in 80% of the positive PBD infectious mononucleosis (IM). On the other hand, EBV antibodies are preponderant for diagnosis in other clinical manifestations of EBV primary-infection were PBD is not alwasy positive (47%). EBV antibodies of patients with various diseases and antibodies of normal subjects show different profiles. By interpretation of these profiles one discuss the possibility to characterize reinfection, and either latent or active persistant infection.


The relationship of post-stimulus time and interval histograms to the timing characteristics of spike trains. PST (post-stimulus time) and interval histograms computed from recorded spike trains are related to an average timing characteristics of the spike train. The exact nature of this relationship varies with recording parameters, interfering signals, the histogram bin width, and the duration of the measurement interval. This work describes the conditions under which a PST histogram can serve as an unbiased estimate of the ensemble average of a spike train's intensity and an interval histogram can serve as an unbiased estimate of the probability density function of the interspike intervals. Simulation studies are used to confirm the validity of the theoretical results. As an example of an application, these results are used to analyze recordings of singleunit activity in the eight cranial nerve.


[Cyclic AMP activation of respiration of the liver mitochondria in different metabolic states]. cAMP 10(-6) activates the liver mitochondria respiration in all the metabolic states and failed to change or increased the phosphorylation rate in the oxidation of saturating concentration of succinate and isocitrate. Preincubation of mitochondria or homogenate of the liver with cAMP is obligatory for this effect. The fraction V of serum albumin and EDTA did not prevent the effect. Noradrenaline enhanced the mitochondrial respiration only in incubation with the homogenate. The effect of noradrenaline and cAMP was not summed up. Probably the noradrenaline effect was mediated through cAMP. The data obtained are against the decisive role of the respiration and phosphorylation uncoupling or the oxidation substrate accumulation and lead to the assumption on the mitochondria enzymes activation.


[Complementary RNA in the chicken sarcoma cells and Rous sarcoma virus]. The subcellular localization in chicken Rous sarcoma of nucleotide sequence, complementary to Rous sarcoma virus RNA was examined by RNA/RNA molecular hybridization. The preparations of radioiodinated virion RNA were annealed with RNAs from different fractions (nuclei, mitochondria, free and membrane-bound polyribosomes) isolated from chicken Rous sarcoma. Formation of RNA-ase resistant hybrids between the viral 125I-RNA and RNA from the mitochondria and membrane-bound polyribosomes was revealed. The latter were characterized by a higher relative redundancy of nucleotide sequences complementary to virion RNA than that in the former, by factor 446. The role of complementary ribonucleotide sequences is discussed.


[Hematopoietic tissue reactions associated with the growth of syngeneic transplanted tumors in mice]. Hemangiopericytoma growth in syngeneic male mice F1 (CBA X C57BL/bj) led to regular hemopoietic changes: an increase in the spleen weight, spleen cell count, in the number of colony-forming units (CFU). It also induced augmentation of myelopoiesis in the spleen and leukocytosis with a sharp increase of segmented granulocytes in the circulating blood characterized as the "leukemoid reaction" syndrome. The latter occurred even when the tumour cells were transplanted into the splenectomized host. Although less marked, this leukemoid reaction was also noted in advanced hepatoma transplanted to syngeneic male mice F1 (CBA X C57BL/6j). No leukemoid reaction was observed in these mice after grafting syngeneic strain of urinary bladder carcinoma.


[Thermal sensitivity of specific transplantation antigens of the tumor-cell membrane]. Study of temperature-sensitivity of the tumour specific transplantation antigen (TSTA) on the cell membrane of SV40-induced tumours and spontaneous hepatoma of inbred Syrian hamsters, as well as in the monkey cells infected in vitro with SV40 virus (ts-mutant) demonstrated high thermolability of TSTA. Heating such cells at 56 degrees C for 30-60 min led to complete loss of their immunogenic activity. Moreover, in the animals immunized with heated tumour cells the test-tumour cell growth was regularly enhanced.


[Participation of hypothalamic adrenergic and cholinergic systems in the regulation of pituitary adrenocorticotropic function]. Experiments were conducted on male rats; a study was made of the activity of pituitary-adrenal cortex system under conditions of electrolyte destruction of various hypothalamic areas. Pharmacological analysis pointed to the irregular distribution in the hypothalamus of the cholinergic and adrenergic systems controlling the secretion of ACTH-glucocorticoids. As suggested, the cholinergic systems are situated in the mammillary region, alpha-adrenergic ones--in the anterior or the posterior regions of the hypothalamus, and beta-adrenergic receptors--in the ventromedial nuclei of the hypothalamus. The latter possibly play an inhibitory role.


[Hyperlipidemia in rats with chronic renal failure]. The character of hyperlipidemia was studied in rats with chronic uremia induced by subtotal nephrectomy--5/6 of the renal tissue was removed. 13 to 30 weeks after this operation the blood serum cholesterol and phospholipid concentration almost doubled. Hyperlipidemia was more pronounced in rats with high azotemia (blood urea nitrogen--BUN). No elevation of serum tryglycerides occurred. Total serum beta- and pre-beta-lipoproteins determined nephelometrically increased significantly only with the BUN level of over 80 mg%. Lipoprotein disc electrophoresis of the serum in rats with uremia demonstrated a distinct rise of alpha-lipoproteins and a slight--of beta-lipoproteins; postheparin lipolytic activity of the plasma was normal. Experimental rats displayed massive proteinuria, but hypoproteinuria was insignificant.


[Beta-adrenoreceptors on the surface membranes of lymphocytes and macrophages]. In experiments in vivo and in vitro on sensitized and intact guinea pigs and Wistar rats the effect of beta-adrenergic stimulants (adrenaline and izoproterenol) and a beta-adrenergic blocker (propranalol) on lymphocytolysis and the reaction of macrophage adherence inhibition was studied. Adrenaline and izoproterenol were shown to inhibit the antigen interaction with both sensitized and intact cells. Restoring the sensitivity of cells to the antigen, propranalol destroys the defensive action of adrenaline and isoproterenol.


[Characterization of the viral-type nucleotide sequences detected in the RNA of human leukaemic cells by probes of murine and simian origins (author's transl)]. Molecular hybridization techniques were used for searching nucleic acid sequences homologous to murine and simian oncornaviral genomes in the RNA of various categories of human leukaemic cells. We report attempts to characterize the sequences that were detected in defined categories of leukemias. It is shown that: i) although there is some correlation between the two probes used in our study with regard to the positivity or the negativity of the tests for the same leukaemic cases, there are also some discrepancies since some of the cases are positive with one probe and negative with the other; ii) the common sequences of the two probes when isolated, were ineffective to detect any complementary sequences in leukemic cases which were scored as positive when the entire probes were used; iii) the complementary sequences detected in three positive cases of acute myelogenous leukemias by a recycled probe are additive and therefore most probably distributed along the viral genome.


[Synovialo-sarcoma of the laryngopharynx: electron microscopic study (author's transl)]. The authors report a new case of synovialo sarcoma of the laryngo-pharynx by optic and electron microscopy. Ultrastructural features were similar to those of joint synovialo-sarcoma, i-e, showing glandular like and spindle cell componments. Both of these shared several ultrastructural characteristics, favouring the hypothesis of a common origin. The site and the fine structure of this tumor are discussed according to the literature.


Human studies following models of tumorigenesis by DNA tumor viruses in animals. The discovery of virus-specific messenger RNA in virus-induced animal tumors has led to the search for messenger RNA in human tumors that can be hybridized with the DNA of known oncogenic viruses. Attention has focused on the adenoviruses, which have produced cancer in laboratory animals and are widespread in man, and on three papovaviruses that have been isolated in human disease and which are oncogenic in hamsters. In other research, the association between human infection with herpesivurs type 2, which is likewise oncogenic in hamsters, and invasive carcinoma of the cervix is being examined. An experimental vaccine is being developed, and nonhuman primate models are being studied as part of this work. Epstein-Barr virus is still another suspected agent of human malignancies, specifically Burkitt's lymphoma and postnasal carcinoma. High prevalence of antigen to hepatitis B virus has been seen to correlate with high incidence of primary liver cell carcinoma, and studies are attempting to elucidate the relationship.


[Subcellular distribution of hypothalamic neurohormones and in vitro stimulation of their release]. Neuronal compartments can be separated by differential spinning or by centrifugation on continuous or discontinuous density gradients. Application of these fractionation techniques to brain structures containing neurosecretory neurons shows that LHRH, somatostatin and a non dopamine prolactin inhibiting factor (PIF) are exclusively recovered from synaptosomal fractions. This indicates that biologically and/or immunologically reactive forms of these hormones are almost entirely concentrated in nerve-endings of neurosecretory neurons. In contrast, other neuropeptides - posterior pituitary hormone, but also TRH, a vasoactive intestinal peptide (VIP), substance P or endorphins - are also found in supernatant fractions. The existence of multiple molecular forms of neuropeptides is likely to explain these differences. Current theories postulate that they are synthetized on ribosomes as precursor forms. Their active structure is only achieved by enzymatic splitting of the pre- or the prohormone within nerve endings. This mode of synthesis is probably common to all neuropeptides, although it has only been well substantiated in a few cases, in particular for the hormones of the posterior pituitary. Thus, the lack of immunologically detectable LHRH or SRIF outside the synaptosomal fraction may reflect masking of the active immunological sites by inert peptide chains associated with prohormonal forms. Fractionation methods can also be applied to physiological or pharmacological experiments. In particular, they permit to characterize, on presynaptic membranes of neurosecretory neurons, specific receptors to neurotransmitters involved in the control of neurohormone secretion. Interaction of dopamine and acetylcholine with LHRH and CRF release are presented as examples of such applications.


[Specific role of sex steroids on the pituitary level. Sex steroids and anterior pituitary secretion]. Although sex steroids were known to play a role in the control of LH, FSH, TSH and prolactin secretion, in vivo experiments could not discriminate between hypothalamic and pituitary sites of action. In this study, the specific action of sex steroids at the anterior pituitary level could be achieved using rat adenohypophyseal cells in primary culture. While estrogens stimulated the sensitivity of the LH and FSH responses to LHRH, androgens had differential effects on the secretion of the two gonadotropins: marked inhibition of LH and stimulation of FSH secretion. Progesterone, on the other hand, while having no effect in the absence of estrogens, could reverse the stimulatory effect of estrogens on LH release while it led to a stimulation of FSH secretion. Estrogens and thyroid hormone exert respective stimulatory and inhibitory effects on TSH secretion by a direct action at the pituitary level. These effects appear to be mediated changes of the level of adenohy-pophyseal TRH receptors. A close correlation was observed between the specificity of binding of the dopamine agonist (3H)dihydroergocryptine and the control of prolactin release in cells in culture, thus supporting the physiological importance of the dopamine receptor in the control of prolactin release. The high degree precision of this system permits assessment of activity of not only dopamine agonists and antagonists, but also of compounds having mixed agonist-antagonistic activity. Preincubation of anterior pituitary cells with 17beta-estradiol not only stimulated basal and TRH-induced prolactin release but, more unexpectedly, led to an almost complete reversal of the inhibitory effect of dopamine agonists on prolactin secretion. Besides its own interest, the adenohypophyseal cell culture system could well be used as a model system for study of the interaction between estrogens and dopaminergic action.


Interaction of alpha-melanotropin with central dopamine systems: role of hormonal state and molecular structure. The tubero-infundibular and nigrostriatal DA neurone systems of rats respond to systemic (i.p.) injection of alpha-MSH (2-100 microgram/kg). The response of the tubero-infundibular (arcuate) DA neurones, an increase in cellular fluorescence intensity which can be interpreted as a sign of increased neuronal activity, is essentially the same in males, estrogen-progesterone-pretreated ovariectomized females and hypophysectomized males, whereas the type of response elicited by alpha-MSH in the nigral DA neurones depends upon the hormonal state of the animal. Differences between the two DA neurone groups exist also with regard to the effects of peptide fragments containing the two active sites of the alpha-MSH molecule. Results of lesion experiments in the lower brainstem (area postrema) and of blockade of muscarinic mechanisms by atropine further point to differences in the mechanisms underlying the peptide effects on the two neurone systems. The reaction of the tubero-infundibular DA system (which controls the pars intermedia of the pituitary) can be considered to reflect the activation of a feedback mechanism on MSH secretion, while the functional counterpart of the changes observed in the nigral system remains unknown at the present time.


[Hypothalamic factors in the human fetal brain: their role in the ontogeny of fetal hypophyseal functions]. The anterior pituitary gland of the human fetus has the ability of synthetizing, storing and secreting hormones early during gestation. The patterns of plasma concentrations of hGH, ACTH, LH and FSH during gestation indicate a maximum of secretion at mid-gestation followed by a progressive decrease of these concentrations until term. In contrast, the secretions of PRL and TSH are moderate at mid-gestation and only increase in the last trimester of gestation. Effective control by the central nervous system (CNS) of the pituitary secretions is still immature at mid-gestation. The presence of releasing factors in the fetal hypothalamus has been established (TRF, LRF, somatostatine) and it was postulated that early in life, relatively autonomous and unrestrained secretion of hypothalamic hypophysiotropic releasing factors occurs and, later in development, there was a maturation of inhibitory or restraining influences mediated via the CNS that modulate the secretion of the fetal adenohypophyseal hormones. Observations made with anencephalic newborns confirm that a functional hypothalamus is necessary for the secretions of each of the hormones of the anterior pituitary gland with the exceptiion of PRL, the secretion of which is normal in anencephaly. Although somatostatin probably participates in the regulation of hGH during fetal life, it appears evident that this regulation can only be fully understood with the existence of a GRF (Growth Hormone Releasing Factor).


The participation of luteinizing hormone releasing hormone (LRH) in the process of sexual maturation. Precious pubertal ovulation and premature steroid-induced gonadotropin surges were triggered in 23-day old rats by unilateral electrolytic lesions placed in the basal hypothalamus. Attempts were made to reproduce both aspects of precocious sexual maturation by repeated injections or prolonged infusion of Luteinizing Hormone Releasing Hormone (LRH) or by repeated administration of the potent and long-acting analog (D-Leu6, des-Gly-NH2(10))-LRH-ethylamide. Neither effect of the brain lesion was reproduced by LRH or its analog. It is concluded that brain lesions advance puberty in this species by mechanisms not involving the discharge of LRH. The role of the adrenal cortex in providing estrogen precursors required for the induction of ovarian gonadotropin receptors is discussed.


Studies on polypeptide receptors a critical view on the mechanism of ACTH action. Recent work on ACTH receptors in adrenal cells and adipocytes is reviewed. It is suggested that ACTH acts on two types of receptor: an adenylate cyclase receptor and another one that helps to guide cAMP into the correct compartment. The introduction of the term holoreceptor to indicate a unit that is recognized by the hormone and is able to produce a measurable effect, and of the terms discriminator, transmitter and effector to indicate the functional subunits of the holoreceptor is suggested as a measure enhancing clarity and flexibility of communication in this very complex field.


Effects of releasing factors on hypothalamic neurones. Our previous reports on the effect of locally applied polyeptides such as ACTH, the releasing factors TRF and LRF on single unit activity of hypothalamic neurones have been confirmed and extended by several workers. These polypeptides also exhibit behavioural effects. The concept that such hormonal polypeptides may exert neurotransmitter as well as neuromodulator function has gradually become acceptable.


Mechanism of alpha-melanotropin action.

Mechanism of alpha-melanotropin action. A structure-function study of alpha-melanotropin has shown that this tridecapeptide consists of two message sequences, (-Glu)-His-Phe-Arg-Trp- and -Gly-Lys-Pro-Val-NH2, and a potentiator sequence, Ac.Ser-Tyr-Ser-Met-(Glu-), when acting on its melanophore receptors. The key elements of the message, -Phe-Arg- and -Lys-Pro-, do not correspond exactly to those responsible for eliciting the effect in other tissues. It appears that alpha-MSH contains more information than would be necessary to interact with only one complementary receptor site; therefore, the topography of the hormone exposed to the binding site may be different on contact with the receptors of different target cells. To further investigate this aspect, new methods for the isolation and characterization of functional receptors must be developed. We are investigating the use of chemically well-defined, biologically active, covalent hormone-macromolecule complexes for this purpose. Another approach utilizes model receptors with a recognition pattern similar to that of the biological receptor, as described in this communication for certain highly specific antibodies.


Depression of evoked potentials in brain slices by adenosine compounds. 1 A study has been made of the action of adenosine on surface slices of guinea-pig olfactory cortex in vitro. 2 With extracellular recordings from the pial surface and stimulation of the presynaptic input, the lateral olfactory tract (LOT) generated a monosynaptic negative wave representing dendritic excitatory potentials. This negative wave was depressed by bath application of 1 micron adenosine with increasing effect up to 1 mM. Adenosine 5'--triphosphate (ATP), adenosine 5'-monophosphate (AMP) and cyclic adenosine 3',5'-monophosphate (cyclic AMP) had similar depressant actions. Adenine and guanosine were very weak depressants. 3 Theophylline concentrations in the range 10 micron to 3 mM progressively antagonized the action of adenosine. 4 Dibuyryl cyclic AMP (100 micron) and agents which increase intracellular cyclic AMP were not depressants, suggesting that the action of adenosine was not cyclic AMP-mediated. 5 Intracellular recordings confirmed the depressant effect of adenosine on excitatory potentials generated by LOT stimulation and also showed that postsynatpic action potentials and the membrane of the soma were unaffected by adenosine. 6 Since presynaptic action potentials were also unaffected by adenosine, these experiments suggest that adenosine reduces excitatory transmission at LOT synapses and fortifies the idea that adenosine has a 'neurohumoral' action.


Morphine analgesia and cerebral opiate receptors: a developmental study. 1 Development of the analgesic response to morphine and ontogenesis of central opiate receptors were analyzed in rats 5 to 120 days old. 2 The analgesic effect of morphine increased until day 15, after which it decreased to reach a plateau at about day 30. With phenoperidine, on the other hand, the analgesic effect increased until day 15, remained constant between day 15 and day 30 after which it decreased slowly. 3 The ratio of the amounts of morphine in blood over those in brain increased about 3 fold between day 15 and day 30. 4 Opiate receptors were detected in the brain of newborn rats: stereospecific binding of [3H]-naloxone at 10 and 50 nM indicated the presence of low and high affinity binding sites. 5 The number of [3H]-naloxone binding sites increased rapidly during the second and third week after birth. Their affinity for several opiates remained constant throughout development. 6 These results indicate that the analgesic activity of opiates varies with age: until day 15, the analgesic effect of opiates increases in parallel with the number of opiate brain receptors. Then, the formation of the blood brain barrier introduces an additional step in the regulation of opiate activity.


The effect of different calcium concentrations on the inhibitory effect of presynaptic alpha-adrenoceptors in the rat vas deferens. The effects of different calcium concentrations on the twitch-inhibitory potency of clonidine and of guanethidine have been examined in the rat isolated vas deferens. Vasa deferentia did not respond to field stimulation at 0.3 Hz in Krebs solution containing 0.325 mmol/1 of calcium. Increasing the calcium concentration to 5.2 mmol/1 caused a concentration-dependent increase in size of the twitch response. The twitch inhibitory potency of clonidine was inversely proportional to the calcium concentration. The inhibitory effect of guanethidine was not influenced by changes in calcium concentration. The results show that the modulation of motor function by presynaptic alpha-adrenoceptors in the rat vas deferens is calcium-dependent.


Comparison of firing patterns and sensory responsiveness between supraoptic and other hypothalamic neurons in the unanesthetized sheep. Adult Southdown ewes were surgically prepared with pituitary stimulating electrodes, carotid and jugular cannulae, and a cranial platform-cylinder arrangement for chronic single unit recording. Isolated neurons (n = 112) in the region of the supraoptic nucleus (SON) were identified by pituitary stalk stimulastion as AD + (antidromically invaded) SON neuroendocrine cells (n = 75) or AD--(not antidromically invaded) SON neurons (n = 37). Spontaneous firing pattern distribution and sensory evoked behavior of these SON region neurons were compared with activity recorded from 112 randomly located non-identified neurons of extra-SON areas of the hypothalamus. Spontaneous discharge activity was categorized into six distinct firing pattern types: continuously active slow (CAS), continuously active fast (CAF), continuously active bursting (CAB), continuously active regular (CAR), low frequency bursting (LFB), and high frequency bursting (HFB). These 6 firing pattern types were characterized by computer analysis and their mean order independent statistical parameters compared. Bursting discharge patterns (LFB, HFB, and CAB) were compared with respect to mean burst duration, burst mean firing rate, and interburst interavls. Ninety-three per cent of all neurons maintained a stable discharge pattern in the absence of apparent stimuli. Occasionally CAS and CAF neurons spontaneously generated spike clusters sufficient to give the transient appearance of a bursting discharge pattern and LFB neurons lapsed spontaneously into CAS acitivity. All 6 firing pattern types recorded from non-identified extra-SON neurons were also recorded in the SON region. However, spontaneously discharging AD+ SON neurons exhibited only continuously active slow (CAS), continuously active fast (CAF), and low frequency bursting (LFB) activity. The total absence of high frequency bursting (HFB), continuously active regular (CAR), and continuously active bursting (CAB) patterns of discharge from AD+ SON neurons suggests that AD- SON neurons exhibiting these firing patterns may function as interneurons, pacemaker neurons, or receptor neurons. A significant number of LFB discharging neurons were recorded in widespread extra-SON regions of the hypothalamus, indicating this discharge pattern may not be unique to magnocellular neuroendocrine cells. AD+ SON LFB neurons sampled in this study demonstrated a significantly longer mean interburst interval (20.86 sec) compared to extra-SON LFB neurons (12.43 sec). No AD+ SON neuron tested was significantly sensitive to non-specific sensory arousal or sleep-waking state changes. In extra-SON areas of the hypothalamus, 11 of 75 neurons tested to sensory arousal and 6 of 19 neurons tested to sleep-waking changes responded with significant changes in mean firing rate (MFR); no significant difference between firing pattern types was demonstrated in arousal or sleep-waking sensitivity...


The auditory midbrain of a marsupial: the brush-tailed possum (Trichosurus vulpecula). A microelectrode survey was made of the midbrain auditory nuclei of the brushtailed possum (Trichosurus vulpecula), a common Australian marsupial. Information was sought on the tuning characteristics of individual neurones, tonotopic organization and mechanisms of sound localization. It was felt that such information would be of use in future studies of the development and evolution of mammalian hearing. Twelve possums were anaesthetized with ketamine and chloralose-urethane, and recordings were made of extracellular unit discharges in the inferior colliculus during monaural and binaural tonal stimulation. The inferior colliculus of the possum consists of a central nucleus - a darkly stained, densely packed group of cells - flanked laterally by an external nucleus with a lower density of paler cells. Tonotopic organization was demonstrated by discretelytuned elements in the central nucleus, but was not observed in the external nucleus. In the latter region broad and irregular tuning was commonly seen. Most units in both divisions were influenced by binaural stimuli, with patterns of binaural interaction similar to those observed in the cat inferior colliculus. Cells influenced by changes in the interaural time and intensity difference were commonly observed, but only a subclass of these were suited in sensitivity for sound localization. In general, the midbrain auditory system of the possum was similar in unit discharge characteristics and organization to those of the eutherian mammals commonly studied.


Impaired synaptic potentiation processes in the hippocampus of aged, memory-deficient rats. A series of neurophysiological experiments was performed on the Schaffercommissural system of the hippocampus of aged and young anesthetized Fischer rats. The aged Fisher rats were previously found to exhibit retention performance deficits. No obvious differences were found between aged and young animals in amplitude, latency, stimulation threshold, or wave forms of typical synaptic responses when these were elicited by control (0.3 Hz) stimulation pulses. Further, the temporal curves of facilitation during a paired-pulse series were not different in aged and young animals. However, aged and young synapses showed consistently different responses during repetitive stimulation. Synapses of aged animals were deficient in frequency potentiation processes during 12 Hz stimulation; and the aged animals exhibited a delayed rise of post-tetanic synaptic potentiation following a 5 sec, 100 Hz stimulation train. Moreover, aged synapses 'exhausted' more rapidly during continuous 4 Hz stimulation. Throughout these studies a biphasic pattern of potentiation was observed during repetitive stimulation (brief potentiation, depression, renewed potentiation). Aged animals were deficient primarily in development of the second phase of potentiation. This pattern suggests an age-related impairment of some secondary process of potentiation, leading to an increased tendency to synaptic depression during and after stimulation. The possibility that the impaired hippocampal synaptic plasticity may be related to reported deficient behavioral plasticity in the aged animals is considered.


The effects of 1,25-dihydroxycholecalciferol on embryonic bone in vitro: a biochemical and histological study. Explants from embryonic rat and mouse calvaria were cultivated in the presence of different concentrations of 1, 25-dihydroxycholecalciferol (1, 25-(OH)2D3). The bone resorbing effects of the vitamin D3 metabolite were evaluated by measuring the release of calcium, phosphate, lactate and citrate into the culture medium after 24 h of cultivation. The influence on bone morphology was studied using embryonic mouse radii, in which histological phenomena in the bony, cartilagenous and connective tissue compartments were observed. Both kinds of experiments show that 1, 25-(OH)2D3 has effects on embryonic bone which are typical for high concentrations of parathyroid hormone (PTH). However, the maximal effects on calcium release and on histology are 2.5 times less than those of PTH. In addition, some of the histological features such as the effects on epiphyseal cartilage and on osteoclasts were not observed. 1, 25-(OH)2D3 in concentrations up to 2 X 10(-9) M does not affect basal or PTH-stimulated cAMP levels in embryonic rat calvaria. In a concentration of 1 X 10(-8) M, however, a significant decrease in PTH-stimulated cAMP production was found. It is concluded from these in vitro experiments that (1) 1, 25-(OH)2D3 stimulates bone resorption, and (2) in 1, 25-(OH)2D3-induced bone resorption cAMP is apparently not involved as a second messenger.


The interaction of collagenase with hydroxyapatite and related materials and enzymatic properties of the adsorbed enzyme. The commercial collagenase from Clostridium histolyticum was adsorbed on hydroxyapatite, on bovine femur shaft and on enamel and dentin powders. The substrate specificity of the adsorbed enzyme tested, with chromophore substrate, azocoll and native collagen, differed from that obtained with the soluble enzyme. The adsorption and the substrate specificity was also dependent on the adsorbent used. A pretreatment of hydroxyapatite with chondroitin sulphate, hyaluronate and DNA lowered the adsorption of collagenase. Phosphate ion caused desorption of the enzyme from hydroxyapatite. Sodium fluoride caused partial desorption of the enzyme from hydroxyapatite and enamel and dentin powders. Colangenase adsorbed on root surfaces of teeth liberated hydroxyproline containing material.


Radiation induced free radicals as molecular probes in synthetic apatites. Free radicals generated in synthetic apatitic calcium phosphates by X-ray radiation were investigated by electron spin resonance (ESR) spectroscopy. Among the species stable enough at -188 degrees C to be identified were hydrogen atoms, phosphate radicals, and oxygen anion radicals. The ESR spectra were markedly dependent on the specific surface of the mineral. Oxygen radicals dominated the spectra of low specific surface samples while phosphate radicals were the predominant species at higher specific surfaces. Our studies suggest that the oxygen radicals are more stable in the bulk of the crystal while the hydrogen atoms and the phosphate radicals are stabilized at or near the crystal surface. It was concluded that the surface species are potentially capable of serving as probes of biologically relevant mineral-organic interfaces.


Neuromuscular and cardiovascular depression produced by prolonged exposure to Polymyxin B. It has been reported previously that in cats pre-treatment for 24 to 48 hours with neomycin sulphate resulted in a change in the pattern of neuromuscular block subsequently produced with this antibiotic. The unique characteristics of neomycin-induced neuromuscular block observed during acute exposure gave way to a pattern of block usually observed with tubocurarine. In a similar experiment on six cats we have demonstrated that with polymyxin B prolonged exposure did not result in a comparable time-dependent change in the nature of the neuromuscular block it produced, in terms of train-of-four, tetanic, and post-tetanic behaviour of the neurally-elicited muscle response. The differing characteristics of neuromuscular block seen with various antibiotics, e.g. the aminoglycoside neomycin as opposed to the polypeptide polymyxin B, during acute and sub-chronic exposure is stressed. Previously observed ability of 4 aminopyridine 0.6 mg/kg to reverse the neuromuscular and cardiovascular depression during acute exposure to polymyxin B persisted in prolonged exposure.


Purification and some properties of NAD-degrading purine nucleosidase from Aspergillus niger. An enzyme which degrades NAD at the adenine-ribose linkage has been purified from the mycelial extract of Aspergillus niger. NADP, deamido-NAD, and purine nucleosides and nucleotides were also susceptible to the hydrolytic cleavage. Pyrimidine- and nicotinamide-ribose linkages were not attacked. The substrate specificity showed that the enzyme may be classified as  a N-ribosyl-purine ribohydrolase (EC 3.2.2.1). The enzyme had a maximum activity in the pH range of 4.0-4.5 toward NAD. The Km values for NAD, 5'-AMP, and inosine were 3.0, 2.9 and 1.6mM respectively.


Coisolation of glycophorin A and polyphosphoinositides from human erythrocyte membranes. The lipid composition of purified erythrocyte membrane glycophorin was measured. Diphosphoinositide, triphosphoinositide, and phosphatidylserine are the major phospholipids in glycophorin preparation. Nearly all of the radioactive diphosphoinositide and triphosphoinositide extracted from erythrocyte membranes by lithium dïodosalicylate are recoverd in purified glycophorin. There appeared to be no significant enrichment of other acidic membrane phospholipids in the protein. The results do not permit a firm conclusion as to whether the polyphosphoinositides are associated specifically with the membrane protein or whether fortuitous binding has occurred during purification.


Preparation of cell populations with stabilized erythropoietic potential from the primitive streak chick blastodisc: some implications for control of gastrulation. Improved methods for preparation from primitive streak chick blastodiscs of cell suspensions capable of forming erythroid cells in culture have been developed. When blastodiscs were preincubayed with hyaluronidase in the absence of collagenase before cell dispersion and a high concentration of methyl-alpha-mannoside was present in all media, the yields of cells were some 10-fold higher than those obtained by former procedures. Cell suspensions obtained consisted almost entirely of viable cells, yielded large numbers of free mature erythrocytes in liquid culture, and formed erythroid colonies and bursts in solidified medium. The capacity to form differentiated cells after resedimenrtation through Ficoll density gradients was partly stabilized. Addition of gee yolk homogenate to the blastodiscs immediately following treatment with hyaluronidase and to all media used thereafter largely stabilized the capacity to form erythroid cells during resedimentation through Ficoll density gradients. Possible relevance of observations made during development of the procedures to the control of onset of cell migration in the process of gastrulation is indicated.


Purification of the heat-stable inhibitor protein of the Ca2+-activated cyclic nucleotide phosphodiesterase by affinity chromatography. The recently discovered heat-stable inhibitor protein of the Ca2+-activated cyclic nucleotide phosphodiesterase (Sharma, R. K., Wirch, E. & Warg, J. H. (1978) J. Biol. Chem., in press) has been purified 238 214-fold from bovine brain extract using an affinity column of the modulator protein--Sepharose 4B conjugate. The purified sample appears to be homogeneous as judged by sodium dodecyl sulphate (SDS) gel electrophoresis. The protein band has a mobility corresponding to that of a polypeptide of molecular weight 68 000. Since the heat-stable inhibitor protein has a molecular weight of 70 000 under nondenaturing conditions, it suggests that it is a monomeric protein. The protein has no inhibitory activity toward the cAMP-dependent protein kinase or protein phosphatase. The purified sample has been tested for various enzyme activities which include ATPase, GTPase, cAMP phosphodiesterase, cGMP phosphodiesterase, 5'-nucleotidase, and protein kinase. None of these activities are exhibited by the purified sample.


A comparison of some immunological characteristics of very low density lipoproteins of normal and hypercholesterolemic rat sera. The immunological characteristics of a very low density lipoproteins (VLDL) from normal and hypercholesterolemic rat sera were compared using polyspecific antisera to VLDL and high density lipoproteins (HDL) and monospecific antisera to apo-B, apo-C, apo-A-I, and apo-E. Ultracentrifugally isolated VLDL from normal serum were studied by immunodiffusion and found to contain both discrete and associated (with apo-B) apo-C and apo-E, probably in the form of lipid-containing lipoproteins. However, immunoelectrophoresis of whole serum revealed only an associated form of the liporpotein having pre-beta mobility (i.e., VLDL), suggesting that the presence of discrete lipoproteins in isolated VLDL, each containing a single apoprotein family, may represent ultracentrifugal artifacts. Ultracentrifugally isolated VLDL from diet-induced hypercholesterolemic rat serum contained only trace amounts of apo-C and large quantities of apo-E, both of which were totally associated with apo-B. VLDL isolated by ultracentrifugation from perfusate of normal and hypercholesterolemic livers contained only associated lipoprotein complexes made up of apo-B, apo-C, and apo-E in the former but only apo-B and apo-E in the latter. These data suggest that normal VLDL are secreted as lipoprotein complexes containing apo-B, apo-C, and apo-E, which may become destabilized in the circulation. However, VLDL from hypercholesterolemic serum shows a marked diminution in the quantity of apo-C as indicated by the relative incorporation of [3H]leucine in vivo and by polyacrylamide gel electrophoresis of apo-VLDL.


Response of the respiratory tract of calves kept at controlled climatic conditions to bovine Herpesvirus 1 in aerosol. Seven experiments with four calves each were conducted in which the calves spent at least four days of adaptation in an environmental chamber and then were subjected to climatic stress in the form of a number of constant ambient temperature and humidity combinations. On the second day of climatic stress the calves were individually exposed to measured numbers of infectious units of bovine herpesvirus 1 (BHV1, virus of infectious bovine rhinotrachetis) in aerosol. The calves were killed seven or eight days later. Mycoplasma were found in some nasal swabs and in one lung. Certain bacteria but no Pasteurella were often isolated from the lungs. Bovine herpesvirus 1 was isolated from chamber air and from most postinoculation nasal swabs, tracheas and lungs. The number of macro- and microscopic lesions did not appear to be influenced by the climatic conditions of the experiments. The histopathological changes in epithelium at all levels of the respiratory tract were described in detail.


Diagnosis of viral agents associated with neonatal calf diarrhea. During this study, 134 samples have been examined for the detection of the viruses associated with neonatal calf diarrhea. The presence of Nebraska viruses (rotavirus and coronavirus) has been demonstrated by using the electron microscope and the fluorescent antibody techniques while the presence of other viruses has been detected by the observation of a cytopathic effect on monolayer cells of calf testis. The Nebraska viruses have been demonstrated in 107 (80%) out of 134 field case specimens. An association of rotaviruses and coronaviruses was found in 58 cases (54%) whilst the coronaviruses and the rotavirus were found singly in 34 cases (53%) and in 15 cases (14%) respectively. Four bovine virus diarrhea viruses, two infectious bovine rhinotracheitis viruses and two enteroviruses have also been isolated in the preceding 107 Nebraska positive specimens. For the detection of the Nebraska viruses, the fluorescent antibody techniques were more sensitive than the electron microscopy. However, those two techniques must be used simultaneously for a better detection of a greatest possible number of cases.


The protective antigens of equine herpesvirus type 1. Equine herpesvirus type 1 was cultivated in swine testis cell cultures and partially purified by differential centrifugation and centrifugation in a linear sucrose density gradient. The viral envelope was separated from the nucleocapsid by treatment with Rexol 25J followed by sucrose gradient centrifugation. The envelope and nucleocapsid preparations were then electrophoresed in polyacrylamide gel after solubilization with sodium dodecyl sulphate. Hamsters were immunized with various preparations of the partially purified virus, including live or inactivated equine herpesvirus type 1 and viral envelope and nucleocapsid, all derived from the Kentucky D strain of the virus. Challenge of the immunized hamsters, with a hamster-adapted strain of equine herpesvirus type 1 demonstrated protection only in those animals which had been vaccinated with envelope-containing materials. When vaccination was carried out with fractions of electrophoresed envelope or nucleocapsid, protection was induced only by polypeptides of high molecular weight containing a glycoprotein component of the envelope of equine herpesvirus type 1.


A complement fixing antigen in the livers of birds infected with an avian leukovirus (erythroblastosis virus). Extracts of erythroblastosis affected livers from which much of the structural protein had been removed were prepared by acid denaturation. These extracts contained complement fixing "soluble" antigen in a form which was not associated with viral or subviral particles. Ultracentrifugation can be of assistance in separating complement fixation activity from contaminating material in these extracts.


Duration of active and colostrum-derived passive antibodies to bovine viral diarrhea virus in calves. Duration of active and colostrum-derived passive antibodies to bovine viral diarrhea virus was studied in 14 calves. Five calves born with actively induced antibodies to bovine viral diarrhea virus retained high titers during the year of observation. Colostrum-derived antibodies to bovine viral diarrhea virus in nine calves declined at an expected rate for the first four to six months of age. However, titers of six of these calves increased at five to eight months of age and either remained constant or increased through one year of age. Bovine viral diarrhea virus antibody titers of the other three calves declined at a constant rate to less than 1:4 by nine to 12 months of age.


Dissociation of cyclic GMP synthesis from cholinergic-stimulated secretion of protein from rat exocrine pancreas. The phosphodiesterase inhibitors RO-201724/1 and 1-methyl-3-isobutylxanthine (MIX) stimulate a rapid increase in cyclic GMP content in rat pancreas; the latter agent also potentiates the stimulatory effect of carbachol on cyclic GMP synthesis. However, neither RO-201724/1 nor MIX alter basal secretion of 3H-labeled protein, nor do they affect the secretory response to carbachol used in either suboptimal or optimal concentrations. MIX as well does not alter the rate at which carbachol stimulates pancreatic enzyme release. The ability of carbachol to increase cyclic GMP synthesis is lost if extracellular calcium concentration is lowered to 0.05 mM; at this calcium concentration, however, the muscarinic agent still elicits a marked secretory effect. The dissociation between cyclic GMP synthesis and the secretory response suggests that the cyclic nucleotide does not play a major role in the stimulus--enzyme secretion coupling phenomenon of the exocrine pancreas.


Validation of a single-compartment approach to the calculation of secretion rates of ACTH. A single-compartment model used in this laboratory for continuously calculating ACTH secretion rates from measured plasma ACTH concentrations has been tested for its ability to follow changing rates of ACTH entry (rapid departure from steady state). ACTH was infused at known moderately high but physiological rates into anaesthetized dogs (Nembutal). Under such conditions endogenous secretion is initially less than 5% of infused rates. Orthogonal polynomials (ACTHt) were fitted to plasma ACTH vs. time data. Then secretion ratet = (ACTHt X MCR) + (dACTHt/dt X V) where it was previously shown that the metabolic clearance rate of ACTH (MCR) lacked significant inter-animal or concentration-dependent variation, and its distribution volume (V) was also constant. The calculated ACTH entry rate curves (a) followed a 10-fold increase in infusion rate over 4 min and subsequent rapid decline with a lag of only about 1 min and, despite some blurring, gave an integrated response equal to 94 +/- 5.5% of the known signal, and (b) followed a sinusoidal change in infusion rate (amplitude, 1.7 X base rate; period, 40 min) with a few percent error and negligible lag. These signals imitate (a) an abrupt stress response, and (b) other rapid departures from steady state.


Mediation of isoproterenol-induced thirst in rats by beta2-adrenergic receptors. Isoproterenol-induced thirst in rats has been attributed to the activation of beta-adrenergic receptors. Since these receptors can be further differentiated pharmacologically into beta1 and beta2 types, experiments were performed using several beta-adrenergic agonists and antagonists to determine the receptor type initiating the isoproterenol-induced thirst. The beta1- and beta2-adrenergic antagonist, d,l-propranolol (1 mg/kg, ip), blocked the increase in water intake usually accompanying acute subcutaneous administration of isoproterenol (25 microgram/kg) to female rats. Since l-propranolol is known to stabilize membranes and to possess anesthetic-like properties, d-propranolol was also used. This isomer has little beta-adrenergic-blocking activity but possesses anesthetic-like activity. Administration of d-propranolol (1 mg/kg, ip) failed to affect the drinking response to acute administration of isoproterenol (25 microgram/kg). Practolol (125 mg/kg), a beta1-adrenergic antagonist with little anesthetic properties, also had no effect on water intake of isoproterenol-treated rats. Butoxamine, a selective beta2-adrenergic antagonist, attenuated the drinking response to isoproterenol. Salbutamol (150 microgram/kg), a beta2-adrenergic agonist, mimicked the effect of isoproterenol on water intake. These results are consistent with the suggestion that beta2-adrenergic receptors mediate the isoproterenol-induced thirst in rats.


An investigation of the activity of opiate drug receptors located on frog skeletal muscle fibre membrane. Extracellular and intracellular microelectrode studies were conducted to test the actions and interactions of opiate agonists, antagonists, and procaine on action potentials in frog sartorius muscles. Extracellular studies showed that morphine, methadone, propoxyphene, and procaine all depressed action potential production. Low concentrations of naloxone or naltrexone antagonized the excitability depression produced by the three opiate agonists but not the depression produced by procaine. Intracellular studies revealed that certain concentrations of the opiate agonists produced a biphasic decline in the stimulus-induced increase in sodium conductance (gNa). Naloxone or naltrexone antagonized only the second phase of this decline. These results show that part of the excitability depression produced by opiate agonists is due to an action on opiate drug receptors.


Effect of estrogens on apomorphine-induced circling behavior in the rat. The effect of administration of estrogens parenterally for 1 week was tested on apomorphine-induced circling in a group of castrated female rats with a lesion of the left entopednucluar nucleus. We observed a significant decrease in the number of turns per minute in estrogen-treated animals as compared with controls. Our tentative explanation is that estrogens decrease the sensitivity of dopamine receptors in the striatum.


Multiple sclerosis treated with antithymocyte globulin--a five year follow-up. Multiple sclerosis patients treated with antithymocyte globulin (ATG) were re-evaluated after five years. No long term benefit was found. Notably, the group of patients with an elevated gamma globulin to total protein ration in their C.S.F. and who did particularly well after treatment with ATG also failed to show any long term benefit. Few long term detrimental effects of ATG immunosuppression were identified. The implications of the results are discussed as they relate to the use of immunosuppression in multiple sclerosis.


The role of radiation therapy in the treatment of glomus jugulare tumors. The records of 14 patients who received irradiation for incompletely excised, inoperable or recurrent glomus jugulare tumors were retrospectively reviewed. Ages ranged from 12 to 66 years, and the male to female ratio was 1:3. With a follow-up time of 1.3 to 17.2 years (mean of 7.7 years), 11/14 remain clinically disease-free. Doses of at least 4000 rad are shown to be effective in controlling glomus jugular tumors.


Postirradiation sarcoma (malignant fibrous histiocytoma) of axilla. A case is reported of a patient who developed a histologically unusual sarcoma in the axilla and chest wall 8 years after receiving radiation therapy (6500 rad) for carcinoma of the breast. This sarcoma showed light- and electron-microscopic features of a malignant fibrous histiocytoma, a tumor not documented among 24 previously reported cases of postirradiation sarcoma following the diagnosis of breast carcinoma. In addition, the literature is reviewed and discussed regarding postirradiation sarcoma in general following breast carcinoma.


Cell-mediated immunity to varicella zoster virus in children being treated for cancer. Stimulation of lymphocytes by varicella-zoster virus (VZV) antigen was measured in vitro for 37 healthy adults and children and for 35 children who developed herpes zoster or varicella while receiving anticancer therapy. For 35 of the control group the history of varicella infection correlated with the in vitro response of lymphocytes to VZV antigen. A specific lymphocyte response was observed following 32 of the 37 episodes of VZV infection in the 35 children with malignancy. Re-evaluation 7--12 months following the acute infection revealed that the in vitro response to VZV antigen was lost in six of 19 patients who remained in remission and in all patients who relapsed and received reinduction therapy. Although there was some suggestion that development of a cell mediated response early in the infection decreased the incidence of skin dissemination of the virus in herpes zoster, no consistent correlation could be made between lymphocyte responsiveness in vitro and dissemination of disease or the duration of the appearance of new lesions.


Ultrastructure of primitive neuroectodermal neoplasms of the central nervous system. The ultrastructure of one spinal and five cerebral neoplasms diagnosed by light microscopy as primitive neuroectodermal tumors supports a cell population consisting largely of poorly differentiated neuroepithelial cells. The most unique ultrastructure feature was the presence of annulate lamellae in four of the six cases. Glial cells in the neoplasm were not unequivocally of neoplastic origin and were possible reactive. There was no evidence of neuroblastic or neuronal elements, although there was frequently focal early neuroblastic differentiation by light microscopy. Although we have seen neoplasms which are clearly neuroblastic, these particular tumors are not purely neuroblastic and should not be classified as neuroblastomas.


Glycosaminoglycan-synthetic activity of pleomorphic adenoma. Adenoid cystic carcinoma and nonneoplastic tubuloacinar cells of the salivary gland. An analysis was carried out on glycosaminoglycan produced in pleomorphic adenoma, adenoid cystic carcinoma, sialadenitis and normal tissue of the salivary gland. After incubation of the tissue segments in a medium containing 35SO4, a radioautograph of the tissue section was made to observe the localization of 35SO4 incorporation, and 35S-labelled materials were purified from the tissues, and analyzed. High 35S-radioactivity was observed in the ductal cells of the inflammatory gland tissue and in the acinar cells of normal palatinal gland, but little radioactivity was observed in the interstitial components in these tissues, and the amount of 35SO4 incorporated in the tumor cells was also significant. Eighty to 90% of the 35S-radioactivity incorporated could be detected as 35S-glycosaminoglycans in all tissues except for the normal palatinal gland, which contained a large amount of 35S-sulfated glycoprotein. No significant difference in the synthetic activity of 35S-glycosaminoglycans and in their components were observed between nonneoplastic and neoplastic cells. These results suggest that glycosaminoglycan-producing cells in pleomorphic adenoma as well as in adenoid cystic carcinoma are derived from the tubuloacinar cells of the salivary gland.


Malignant mixed tumor of the vagina probably arising in mesonephric rests. An exceedingly rare tumor arising in the upper lateral vagina is described histologically, histochemically, and ultrastructurally. It is compared to the lesion reported by Okagaki et al. which is considered to represent the same type of tumor. Comparison with synovial sarcomas indicates that the lesion is similar, but the reasons why it should not be so classified are discussed. Evidence is presented for the origin of these tumors in mesonephric rests (Gartner's duct).


Clear cell sarcoma of tendons and aponeuroses: a comparative study of 13 cases with a provisional subgrouping into the melanotic and synovial types. Electron microscopy of two cases of clear cell sarcoma of tendons and aponeuroses showed different fine structures. On the basis of these differences a proposed division is made of melanotic and synovial types of the tumor. A subsequent comparative histological study carried out on these tumors showed that there were histological criteria to separate the melanotic and synovial types of sarcoma. Using these histological criteria eleven further cases of clear cell sarcoma in the files of the Tumor Registry were subdivided. Review of the Tumor Registry material showed that eight were melanotic type tumors and one was a further synovial type tumor. Using these histological criteria there are two further cases in the Tumor Registry which are not clearly classifiable into either group. The present observations indicate that the majority of the so-called clear cell sarcomas are in reality "melanomas of soft parts" with a minority of this type of tumor being of "synovial type".


Association of carcinoma of the breast with adenosquamous carcinoma of endometrium. Numerous investigators have alluded to an association of cancer of the endometrium and breast. There is no available information relating the histologic forms of cancer of the endometrium to breast cancer. We found ten cases of adenosquamous carcinoma of the endometrium in one tumor registry in a ten year period, 1967-1977. Five of these cases also had breast carcinoma. In three of these cases the breast carcinoma was discovered two to three years after hysterectomy; in one case both tumors were discovered at the same time, and in one case the breast carcinoma antedated the diagnosis of endometrial malignancy by approximately one year. These observations, although from a small number of cases, warrant an expansion of this study in order to determine whether patients with adenosquamous carcinoma of the uterus have a higher risk of developing breast cancer.


Undifferentiated (embryonal) sarcoma of the liver: report of 31 cases. Thirty-one cases of undifferentiated (embryonal) sarcoma of the liver are presented. The tumor is found predominantly in the pediatric age group, the majority of patients (51.6%) being between 6 and 10 years of age. An abdominal mass and pain are the usual presenting symptoms. Radiographic examination is nonspecific except to demonstrate a space-occupying lesion of the liver. The tumors are large, single, usually globular and well demarcated, and have multiple cystic areas of hemorrhage, necrosis, and gelatinous degeneration. Histologic examination shows a pseudocapsule partially separating the normal liver from undifferentiated sarcomatous cells that, near the periphery of the tumor, surround entrapped hyperplastic or degenerating bile duct-like structures. Eosinophilic globules that are PAS positive are usually found within and adjacent to tumor cells. Areas of necrosis and hemorrhage are prominent. The prognosis is poor, with a median survival of less than 1 year following diagnosis.


Chemoimmunotherapy of small cell bronchogenic carcinoma. Thirty-one patients with small cell bronchogenic carcinoma were treated with a regimen of cyclophosphamide, Adriamycin and vincristine. Radiotherapy was given to patients with limited disease. Nonspecific immunotherapy consisting of BCG scarification was administered after each chemotherapy course. The results of treatment for this group were compared with those for a group of 45 patients treated similarly but with no immunotherapy. Therapeutic results, expressed in length of survival and rate of response, were similar. Myelosuppression was not modified by the addition of BCG scarification. This study showed no benefit from the use of such nonspecific immunotherapy.


Chylous ascites following retroperitoneal lymphadenectomy: report of 2 cases with guidelines for diagnosis and treatment. Two cases of chylous ascites following retroperitoneal lymphadenectomy are presented. The first case, a 3-year-old girl, undersent a right radical nephrectomy and retroperitoneal lymph node dissection for Wilms' tumor. Chylous ascites developed postoperatively and resolved after 3 months of supportive therapy. The second case, a 45-year-old woman, underwent a right radical nephrectomy and retroperitoneal lymph node dissection for renal carcinoma. Chylous ascites occurred postoperatively and resolved after a single paracentesis. Diagnosis, evaluation, and therapeutic modalities are outlined and the literature is reviewed.