Glutathione: interorgan translocation, turnover, and metabolism. Glutathione is translocated out of cells; cells that have membrane-bound gamma-glutamyl transpeptidase can utilize translocated glutathione, whereas glutathione exported from cells that do not have appreciable transpeptidase enters the blood plasma. Glutathione is removed from the plasma by the kidney and other organs that have transpeptidase. Studies in which mice and rats were treated with buthionine sulfoximine, a selective and potent inhibitor of gamma-glutamylcysteine synthetase and therefore of glutathione synthesis, show that glutathione turns over at a significant rate in many tissues, especially kidney, liver, and pancreas; the rate of turnover in mouse skeletal muscle is about 60% of that in the kidney. Experiments on rats surgically deprived of one or both kidneys and treated with the gamma-glutamyl transpeptidase inhibitor D-gamma-glutamyl-(o-carboxy)phenylhydrazide establish that extrarenal gamma-glutamyl transpeptidase activity accounts for the utilization of about one-third of the total blood plasma glutathione. Normal animals treated with the transpeptidase inhibitor excrete large amounts of glutathione in their urine. They also excrete gamma-glutamylcysteine, suggesting that cleavage of glutathione at the cysteinylglycine bond may be of metabolic significance. The present and earlier findings lead to a tentative scheme (presented here) for the metabolism and translocation of glutathione, gamma-glutamyl amino acids, and related compounds.


Prevention of genetic anemias in mice by microinjection of normal hematopoietic stem cells into the fetal placenta. Mice homozygous for mutant genes at the W locus have a marked macrocytic anemia that is fatal in some genotypes. The defect is believed to originate in the developmentally pluripotent hematopoietic stem cell population. Anemia is first grossly manifest on day 13 of gestation, when the liver is the chief hematopoietic organ. The known paucity of blood-forming foci in livers of homozygotes and the limited formation of their erythrocytes suggested that such fetuses-unlike normal ones-might have conditions favorable for in utero seeding of genetically normal hematopoietic tissue. If this were accomplished before day 13, the anemia might essentially be prevented, or at least substantially mitigated, and normalcy soon achieved by cell selection. This proved to be the case. Allogeneic normal fetal liver cells were microinjected into the blood vessels of the fetal placenta on day 11 of gestation. Of eight mutant homozygotes born from segregating matings, six (four W/W, two W(v)/W(v)) were successfully populated with donor cells. Strain-specific hemoglobin markers demonstrated replacement of the erythroid lineage with the normal type, the rate of substitution being more rapid in the W/W (ordinarily more anemic) recipients. Strain-specific isozyme differences revealed that white blood cells were also replaced. Thus, the initial selective pressure, hence the W-mutant phenotypic lesion, must have occurred at the pluripotent stem cell stage. The animals remained immunologically tolerant of the donor cells and no graft-versus-host reaction occurred. The early introduction of hematopoietic cells differing genetically from all the other tissues of the animal provides possibilities for tracing normal hematopoietic lineages in vivo, for analyzing cell and tissue interactions, such as those between lymphocytes and thymus, and for clarifying the etiology of other blood or immune insufficiencies or malignancies.


Epidermal growth factor stimulation of DNA synthesis is potentiated by compounds that inhibit its clustering in coated pits. We have used inhibitors of receptor-mediated endocytosis to investigate the mechanism and function of epidermal growth factor uptake by cultured cells. When rhodamine-labeled epidermal growth factor is bound to cell surface receptors on confluent monolayers of BALB/c 3T3 cells, it rapidly collects in cell surface clusters and is internalized. The clustering of occupied receptors requires Ca(2+) and is inhibited by primary alkylamines; both of these properties are shared by the enzyme transglutaminase (R-glutaminyl-peptide:amine gamma-glutamyl-yltransferase, EC 2.3.2.13). In Chinese hamster ovary cell extracts, methylamine inhibits 25-50% of the transglutaminase activity with a K(i) of 0.2 mM, and it inhibits the remaining transglutaminase activity with a K(i) of 20 mM. Clustering is almost completely inhibited by 10 mM methylamine. The polypeptide antibiotic bacitracin inhibits clustering of rhodamine-labeled epidermal growth factor or alpha(2)-macroglobulin at 0.7 mM, and it inhibits approximately 40% of the transglutaminase activity in Chinese hamster ovary cells with a K(i) of 0.03 mM. Fluorescent ligands bound to cell surface receptors in the presence of bacitracin form clusters within 30 min after bacitracin is removed from the culture medium. These results indicate that a transglutaminase-like enzyme may be required for the clustering and subsequent internalization of occupied receptors. The effects of 10 mM methylamine and 0.7 mM bacitracin on epidermal growth factor stimulation of DNA synthesis were examined. The stimulation of DNA synthesis by epidermal growth factor was increased 2- to 7-fold in the presence of methylamine or bacitracin. Alone, methylamine or bacitracin increased DNA synthesis 1.1- to 3-fold. The stimulation of DNA synthesis resulting from the simultaneous presence of the hormone and the clustering inhibitor was always greater than the sum of the stimulations produced by the hormone and the clustering inhibitors alone. The potentiation of epidermal growth factor activity by clustering inhibitors suggests that the hormone acts at the cell surface. We propose that rapid internalization of occupied receptors via coated pits may be a mechanism to limit the response to hormones.


Initiation of plasma prorenin activation by Hageman factor-dependent conversion of plasma prekallikrein to kallikrein. Plasma prorenin is an inactive form of renin (EC 3.4.99.19) that can be converted to active renin in acid-treated plasma by an endogenous serine protease that is active at alkaline pH (alkaline phase activation). To identify this enzyme we first tested the ability of Hageman factor fragments, plasma kallikrein (EC 3.4.21.8), and plasmin (EC 3.4.21.7) to activate prorenin in acid-treated plasma. All three enzymes initiated prorenin activation; 50% activation was achieved with Hageman factor fragments at 1 microgram/ml, plasma kallikrein at 2-4 microgram/ml, or plasmin at 5-10 microgram/ml. We then showed that the alkaline phase of acid activation occurred normally in plasminogen-free plasma but was almost completely absent in plasmas deficient in either Hageman factor or prekallikrein; alkaline phase activation was restored to these latter plasmas when equal parts were mixed together. Therefore, both Hageman factor and prekallikrein were required for alkaline phase activation to occur. We then found that, although plasma kallikrein could activate prorenin in plasma deficient in either Hageman factor or prekallikrein, Hageman factor fragments were unable to activate prorenin in prekallikrein-deficient plasma. These studies demonstrate that alkaline phase prorenin activation is initiated by Hageman factor-dependent conversion of prekallikrein to kallikrein which, in turn, leads to activation of prorenin. In this fashion, we have revealed a possible link between the coagulation-kinin pathway and the renin-angiotensin system.


Social gliding is correlated with the presence of pili in Myxococcus xanthus. Myxococcus xanthus, an organism whose motility involves cell interactions, normally bears pili. Myxococcal pili are found only at cell poles, are less than 10 nm in diameter, and may be longer than a cell. Myxococcus has two basic patterns of cell movement, adventurous (A-motility) and social (S-motility). Pili are found to be completely correlated with the presence of S-motility. (The S-motility pattern has many groups of cells, almost no single cells, and is governed by a set of genes called system S.) On the other hand, A-motility is in dependent of piliation. (The A-motility pattern has many single, isolated cells and it is governed by a second set of genes called system A.) Electron microscopic examination of more than 40 genetically different strains shows that all A+S+ (wild-type) and A-S+ strains have pili, but A+S- and A-S- strains lack them. Mutations in four different loci belonging to system S were tested and were found to stop productions of pili: the loci sg1A, sg1B, sg1G, and tg1. When brought into contact with tg1+ cells, cells of a tg1- strain, which lack pili, become phenotypically S+, produce pili, and become S-motile. Both motility and the production of pili are transient when initiated in this way. Thus it appears that pili permit cells that are close to one another to move.


Specific association of neurotransmitter with somatic lysosomes in an identified serotonergic neuron of Aplysia californica. Injection of [3H]serotonin directly into the cell body of the giant metacerebral neuron (GCN), an identified serotonergic cell in the Aplysia cerebral ganglion, revealed a striking association of the labeled transmitter with large lysosomes. Most of the [3H]serotonin in the neuron is sequestered by lysosomes, which may function as storage depots for the transmitter. This localization is specific: [3H]serotonin was not found in similar lysosomes in the perikaryon of R2, an identified Aplysia cholinergic neuron. [3H]Dopamine, [3H]histamine, and [3H]-N-acetylgalactosamine injected into the serotonergic neuron were not localized in lysosomes. Because of this specificity, it may be possible to use lysosomal binding of labeled transmitter as a radioautographic method for identifying the transmitter type of a nerve cell body. These organelles are lysosomal in origin because they contain acid phosphatase activity. Moreover, examination of isolated GCN cell bodies by fluorescence microscopy revealed the presence of abundant autofluorescent yellow pigment characteristic of lipofuscin. We suggest that [3H]serotonin labels organelles containing membranes that once were integral components of the serotonergic vesicle. Lysosomes might accumulate the transmitter with a high degree of specificity because they contain membranes, perhaps recycled from the neuron's synaptic terminals, that retain their ability to concentrate and bind serotonin.


Higher molecular weight forms of immunoreactive somatostatin in mouse hypothalamic extracts: evidence of processing in vitro. Extracts of mouse hypothalamus made in acid/urea containing protease inhibitors were analyzed for somatostatin immunoreactivity after molecular sieve filtration on Sephadex G-50. Higher molecular weight (higher-M(r)) somatostatin-like forms with apparent molecular weights of 15,000, 10,000, and 6000 could be identified, besides the molecular weight 1600 somatostatin. Immunological identities with somatostatin were unambiguously demonstrated by the analysis of the displacement curves in the radioimmunoassay. The M(r) 15,000, 6000, and 1600 species were purified by affinity chromatography on an anti-somatostatin immune serum covalent conjugate with Sepharose used as immunoadsorbant. After disulfide reduction by dithiothreitol, the size of the M(r) 15,000 and 6000 somatostatin-like species was assessed either by molecular sieve filtration or by polyacrylamide gel electrophoresis. The results indicated that the higher-M(r) somatostatin-like species isolated from the hypothalamus did not result from hormone polymerization by means of disulfide interchange. The processing in vitro of the 15,000 higher-M(r) form of somatostatin was achieved by proteolytic enzymes coeluted with this species during the fractionation of hypothalamic extracts. Under neutral pH conditions the intermediary higher-M(r) forms were generated together with the M(r) 1600 somatostatin-like species. This processing activity could be either strongly inhibited at acidic pH or in acid/urea medium or else eliminated by selective immunoadsorption of the 15,000 higher-M(r) form. Neither trypsin nor the gamma subunit of 7S nerve growth factor was able to produce this processing, suggesting that enzymes with other kinds of specificity may be involved. It is concluded that somatostatin biosynthesis in the mouse hypothalamus may occur via a high-M(r) precursor that is processed into intermediary forms leading to the tetradecapeptide hormone.


Intracellular dye-marked enkephalin neurons in the magnocellular preoptic nucleus of the goldfish hypothalamus. A method that combines intracellular recording, dye marking, and immunocytochemistry makes the study of functional and morphological aspects of enkephalin neurons in the magnocellular preoptic nucleus of the goldfish hypothalamus feasible. By use of multiple techniques, enkephalin neurons can be distinguished from other brain cells and can be reconstructed from drawings of serial sections containing the dye-injected opioid cells. These enkephalin cells and their processes measure 14-42 micron in somata diameter and are unipolar, bipolar, or multipolar. Their electrophysiological properties match those of other mammalian and fish magnocellular endocrine cells. This report confirms the one neuron-one hormone (peptide) hypothesis, supports synaptic over electronic coupling between enkephalin and adjacent hypothalamic neurons, and suggests that chemical and functional classification of single electrophysiologically and neuroanatomically studied central neurons can be achieved.


13C nuclear magnetic resonance studies of anaerobic glycolysis in suspensions of yeast cells. Anaerobic glycolysis in Saccharomyces cerevisiae has been studied by 13C NMR at 90.5 MHz. [1-13c]Glucose and [6-13C]glucose were fed to suspensions of yeast cells. Time courses for concentration changes of the starting material, of courses for concentration changes of the starting material, of the intermediate fructose 1,6-bisphosphate (Fru-P2), and of the end products, ethanol and glycerol, have been followed with 1-min time resolution. The glucose uptake was well fitted by a Michaelis-Menten model, assuming competition of alpha- and beta-glucose for the same site. The Km for the uptake was found to be 10 mM for beta-glucose and 5 mM for alpha-glucose. The concentration of Fru-P2 showed an initial oscillation before it reached a co,stant level. The 13C label, introduced only as [-13C]- or [6-13C]glucose, was observed in Fru-P2 in both the C1 and C6 positions, simultaneously. From the relative intensities of the C1 Fru-P2 and C6 Fru-P2 peaks in the presence of [1-13C]- and [6-13C]glucose, in vivo kinetic information was obtained about the aldolase-triosephosphate isomerase triangle. We found that under the conditions of these experiments the ratios of backward to forward velocities through aldolase and triosephosphate isomerase were 0.9 +/- 0.1 and 0.8 +/- 1, respectively, indicating they were close to equilibrium.


Uroporphyrinogen I synthase from human erythrocytes: separation, purification, and properties of isoenzymes. Uroporphyrinogen I synthase [porphobilinogen ammonia-lyase (polymerizing), EC 4.3.1.8] from human erythrocytes was separated into two active protein peaks (A and B on DEAE-cellulose, by ammonium sulfate fractionation, on Sephadex G-100, and on DEAE-Sephadex A-50 with a NaCl gradient. The final purification was 613 and 743 times for A and B, respectively. The corresponding yields were 2.2 and 3.4% Fraction A was separated further into two (A1 and A2) active protein bands and fraction B into three (B1, B2, and B3) on analytical polyacrylamide disc gel electrophoresis. Bands A1 and A2 were identical with B1 and B2; B3 represented a third isoenzyme. Molecular weights (mean +/- SEM), measured by gel filtration and sodium dodecyl sulfate/polyacrylamide gel electrophoresis, were 38,000 +/- 1000 for B1 and 40,000 +/- 1000 for B2 and B3. Isoelectric focusing on 4% polyacrylamide gel separated both fractions A and B into three active protein bands. Maximal activity of the enzyme was found in gel cuts (5-mm) at pH 5.6 for both fractions A and B.


Amplification and characterization of the proline transport carrier of Escherichia coli K-12 by using proT+ hybrid plasmids. A previous report [Motojima, K., Yamato, I. & Anraku, Y. (1978) J. Bacteriol. 136, 5-9) described the characteristics of mutants (proT-) of Escherichia coli K-12 that are defective in proline transport carrier activity. Two hybrid plasmids from the Clarke and Carbon colony bank were found to complement the mutation by conjugation and transformation. Analysis with restriction endonucleases showed that both plasmid DNAs carried the same fragment of the E. coli chromosome. A polypeptide with a molecular weight of 24,000, specifically coded for by the proT+ plasmid, was identified in the cytoplasmic membrane by using a double-isotope labeling method in a minicell system. The strain harboring the proT+ plasmid has 6 times as much proline transport carrier as the wild strain. This amplification of the carrier makes it possible to measure proline binding to the carrier by microequilibrium dialysis. Detailed analysis of binding indicated that the maximal amount of proline bound to the carrier is 0.70 nmol/mg of protein of the cytoplasmic membrane of the amplified strain. From this value and the assumption that the carrier has one binding site per minimal molecular weight of 24,000, the content of the proline transport carrier in the cytoplasmic membrane was estimated to be 1.7%.


Transport of gamma-glutamyl amino acids: role of glutathione and gamma-glutamyl transpeptidase. This work relates to the hypothesis that one of the mechanisms that mediates amino acid translocation across cell membranes involves the action of membrane-bound gamma-glutamyl transpeptidase on intracellular glutathione and extracellular amino acids to form gamma-glutamyl amino acids. According to this idea, the latter are translocated into the cell where the gamma-glutamyl moiety is removed to yield free amino acids. Previous studies in this laboratory showed that intracellular glutathione is translocated out of many cells. We have now directly examined the transport of gamma-glutamyl amino acids into tissues in the mouse by use of the model substrate L-gamma-glutamyl-L-[14C]methionine sulfone. Of 11 tissues examined, only the kidney showed strong and preferential uptake of the substrate. A substantial amount of the administered L-gamma-glutamyl-L-[14C]methionine sulfone was found intact in the kidney; the total uptake of this compound was greater (by about 2-fold) than that of free L-methionine sulfone. Studies with a number of other gamma-glutamyl amino acids and gamma-glutamyl compounds indicate that the kidney has a relatively specific transport system for gamma-glutamyl amino acids. Small but significant amounts of gamma-glutamylmethionine sulfone were found in the liver and pancreas, suggesting that other tissues may also have this system. Transport of gamma-glutamylmethionine sulfone into the kidney was inhibited by inhibitors of glutathione synthesis and of gamma-glutamyl transpeptidase. The results suggest that both the transpeptidase and glutathione may be involved in transport of gamma-glutamyl amino acids.


Structural order of lipids and proteins in membranes: evaluation of fluorescence anisotropy data. The limiting long-time value of fluorescence anisotropy in membranes is correlated with the orientational order parameter, which characterizes the structural anisotropy of membranes. Existing experimental results for diphenylhexatriene in lipid bilayers are evaluated for the order parameter of lipid order. Steady-state measurements of fluorescence anisotropy can provide the order parameter in good approximation. Proteins in a fluid lipid phase increase the lipid order parameter so determined. Upon comparison with the order parameter from deuterium magnetic resonance, it is concluded that proteins increase the order of the surrounding lipids in off-normal directions. Order parameters of protein order obtained from the limiting value of protein fluorescence anisotropy are discussed with respect to the influence of lipid order on protein order.


Quantitative relationship between beta-adrenergic receptor number and physiologic responses as studied with a long-lasting beta-adrenergic antagonist. The aminobenzyl analog of propranolol, 1- (p-amino-alpha,alpha-dimethylphenethylamino)-3-(1-naphthoxy)-2- propanol, was synthesized and found to be a potent beta-adrenergic blocking agent. The beta-adrenergic receptors of cultured rat C6 glioma cells (2B clone) as assessed by [(125)I]iodohydroxybenzylpindolol binding were decreased 50 and >95% after pretreatment with 8 nM and 1 muM aminobenzylpropranolol, respectively. Unlike propranolol, aminobenzylpropranolol displayed a prolonged blockade of receptors that was maintained during several hours of washing. [(125)I]Iodohydroxybenzylpindolol saturation binding experiments in cells exposed to aminobenzylpropranolol and subsequently washed indicated that the compound effectively diminished receptor number with no change in the affinity of the remaining receptors for iodohydroxybenzylpindolol. Aminobenzylpropranolol inhibited catecholamine-stimulated intracellular cyclic AMP accumulation; with increasing blockade, isoproterenol dose-response curves became progressively shifted to the right but the maximal response was unaltered. Aminobenzylpropranolol inhibited the beta-adrenergic contractile response in atria isolated from rats and guinea pigs. Treatment with 0.1 and 10 muM aminobenzylpropranolol produced decreases of 0.5 and 2 orders of magnitude in the contractile potency of isoproterenol. As in glioma cells, aminobenzylpropranolol failed to decrease the maximal response to isoproterenol. The effects of aminobenzylpropranolol persisted during extensive washing of atria (up to 17 hr). Repeated exposures to isoproterenol at concentrations sufficient to produce maximal tension development also failed to alleviate the blockade. The inotropic potency of histamine in guinea pig atria was not affected by aminobenzylpropranolol. These data suggest that catecholamines are capable of eliciting full biological responses in glioma cells and isolated atria even though the great majority of beta-adrenergic receptors are persistently blocked.


Induction of glutamine synthetase in embryonic neural retina: localization in Müller fibers and dependence on cell interactions. The cellular localization of glutamine synthetase [GSase; L-glutamate:ammonia ligase(ADP)-forming), EC 6.3.1.2] induced by cortisol in the neural retina of chicken embryos was investigated by immunostaining with GSase-specific antiserum and indirect immunofluorescence. In organ cultures of retina tissue, and in the retina in vivo, hormone-induced GSase was found to be confined only to the Müller fibers (retinoglia). Also, in mature chicken retina, which contains a very high level of GSase, the enzyme was detected solely in Müller fibers. In short-term monolayer cultures of dispersed embryonic retina cells, there was no GSase induction and no immunodetectable increase in enzyme level. However, when the dispersed cells were reaggregated and they restituted retinotypic cell associations, GSase could be induced and it was localized in Müller fibers. The results suggest that, in addition to the hormonal stimulus, contact-dependent interactions between Müller glia cells and retina neurons are involved in the mechanism of GSase induction in the retina.


Alpha- and beta-adrenergic stimulation of arachidonic acid metabolism in cells in culture. Madin-Darby canine kidney cells (MDCK) synthesize prostaglandin (PG) F(2alpha), PGI(2) (measured as 6-keto-PGE(1alpha)), PGE(2), PGD(2), and thromboxane A(2) (measured as thromboxane B(2)). When incubated in the presence of norepinephrine (6 muM), the syntheses of these arachidonic acid metabolites are stimulated 3-fold. Norepinephrine's effect can be antagonized by the addition of alpha-adrenergic receptor blocking agents (phenoxybenzamine>phentolamine>yohimbine>dibenamine>tolazoline) but not by the beta-adrenergic blocking drug propranolol. Norepinephrine's stimulation is also inhibited by low concentrations of dihydroergotamine, bromocryptine, ergocryptine, and ergotamine. The stimulation of PG synthesis by norepinephrine is reversible, continues during the 24 hr of incubation, and requires the presence of norepinephrine at the receptor site but it is not blocked by the addition of colchicine, cytochalasin B, or cycloheximide. Neither phenoxybenzamine nor ergotamine at concentrations that block norepinephrine's stimulation of PG biosynthesis suppresses the increase in PG synthesis induced by exogenous arachidonic acid, suggesting that the alpha-adrenergic regulation is not occurring primarily at the cyclooxygenase step in the metabolism of arachidonic acid. In mouse lymphoma cells (WEHI-5), low concentrations of isoproterenol or norepinephrine stimulate the synthesis of thromboxane, an effect that can be blocked by the addition of propranolol but not by relatively high concentrations of phenoxybenzamine or ergotamine. Taken together, these results suggest that alpha-adrenergic receptor stimulation promotes the deacylation of phospholipids by MDCK cells whereas beta-adrenergic mechanisms lead to activation of similar pathways in WEHI-5 cells.


Cellular site of gastric acid secretion. Isolated gastric glands of the rabbit were examined both with differential interference-contrast microscopy and with electron microscopy to describe the morphologic correlates of acid secretion. Stimulation of the glands with histamine resulted in the development of intracellular spaces within the parietal cells. A similar transformation was produced by addition of 1 mM aminopyrine, whether the weak base was added in the presence of normal-K+ (5.4 mM) or high-K+ (108 mM) solutions. The intracellular space was compatible with the expanded canaliculus described in stimulated parietal cells. Confirmation that the space produced by histamine is the site of acid secretion was gained by combining fluorescence and interference-contrast methods in the presence of the dye acridine orange, which displays a pH-dependent metachromasia in its emission spectrum. Human gastrin I resulted in an observable discharge of peptic granules.


Effects of D- and L- enantiomers of adenosine, AMP and ADP and their 2-chloro- and 2-azido- analogues on human platelets. Aggregation of human platelets by ADP and the inhibition of this effect by adenosine are apparently mediated by different receptors. One of the criteria for receptors is that they show stereospecificity for their ligands. We have synthesized L-enantiomers of D-adenosine, AMP and ADP, together with their corresponding photolysable 2-azido analogues so that platelet receptors could be tested for stereospecificity. All of the L-enantiomers were completely inactive as aggregators or inhibitors of platelet function. None of the L-enantiomers changed levels of platelet cAMP. 2-Azido-L-adenosine, AMP and ADP are proposed as useful controls in photoaffinity experiments for non-specific labelling.


Membrane potential and ion concentration stability conditions for a cell with a restricted extracellular space. For an isolated membrane, the resting (zero current) potential is stable is the slope conductance is positive, and is unstable if the slope conductance is negative. Recent work suggests that the properties of many preparations are influenced by the presence of an extracellular space that is not in good diffusive contact with the bulk extracellular fluid. Ionic current flow across the membrane changes the ion concentrations in this space. These concentration changes affect the stability of the membrane potential. Even if the slope conductance is negative, the presence of the extracellular space can confer stability on the resting potential. Conversely, even if the slope conductance is positive, the extracellular space can produce instability of the resting potential. Evaluation of the relevant parameters for cardiac Purkinje fibres, from published experimental data, suggests that concentration changes in the extracellular space may play a significant role in determining when an action potential is initiated.


Oncofoetal antigens of human trophoblast. Rabbits immunized with human trophoblast cell membranes produced antibodies that were detected, by immunofluorescence, to react with normal human tissues, and, by complement-mediated cytotoxicity, with several transformed human cell lines. Absorption with trophoblast abolished all of these reactions, whereas multiple absorptions with lymphocytes, liver or kidney failed to remove reactivity with either trophoblast or certain transformed cells. To further identify the antigens responsible for these antibodies, rabbits were immunized with a chromatographed fraction of deoxycholate-solubilzed membranes prepared from KCl-extracted, ultracentrifuge-prepared trophoblast microvilli. The resultant IgG antibody reacted specifically with syncytiotrophoblastic membranes in sections of human placentae, in addition to recognizing the membranes of viable Chang liver, AV3, HEp-2, Sw/156 (kidney) and Sw/527 (breast) cells. That normal tissues, baboon or monkey placentae, and HeLa or Daudi cell lines did not react with this antibody, indicates the presence of species- and organ-specific antigens in human trophoblast, as well as the existence of trophoblast cross-reactive antigens on some transformed cells. The selective localization of these antigens at the interface of the materno-foetal graft suggests that they function biologically in the host-parasite relation of human pregnancy; their appearance on many transformed cells implies a similar function in the host-parasite relation of some human cancers.


The annual testicular cycle in an equatorial colony of lesser rock hyrax, Heterohyrax brucei. Adult males from a colony of lesser rock hyrax found near the equator in Kenya exhibited an annual cycle of testicular activity characterized by intense spermatogenesis and elevated androgen status from May to July. Average masses of testes and seminal vesicles taken in these months were almost fourfold greater than those from September to January. During the months of peak testicular activity average diameters of Leydig cells and seminiferous tubules were increased by approximately one half and total tubule length was doubled, compared with values for the quiescent months. Variable testicular development occurred during transitional intervals preceding and following peak testicular activity. From February to Aril thickening of the seminiferous epithelium and appearance of spermatozoa in the caput epididymidis signalled re-establishment of sperm production. In August shedding of germinal cells from the epithelium heralded impending failure of spermatogenesis. Evidence of an annual testicular cycle contradicted the prevalent belief that equatorial hyrax breed all year and suggested that the testicular cycle is a conservative element of hyracoid reproductive strategy.


The action of substance P on neurons of the myenteric plexus of the guinea-pig small intestine. Extracellular and intracellular recordings were made in vitro from single neurons of the myenteric plexus of the guinea-pig small intestine. Synthetic substance P was applied to the neurons by means of the perfusing solution or by electrophoresis from micropipettes. Extracellular recording showed that substance P (100 pm-30 nm), applied by perfusion, increased the firing rate of myenteric neurons. Intracellular recording indicated that perfusion with substance P caused a dose-dependent membrane depolarization which was unaffected by hexamethonium, hyoscine, naloxone or baclofen. The depolarization was also evoked by electrophoretic application of substance P. It was associated with an increase in membrane resistance, augmented by membrane depolarization and reduced by membrane hyperpolarization. The relation between the substance P reversal potential and the logarithm of the extracellular potassium concentration was linear with a slope of 54 mV/log10[K+], which indicates that substance P inactivates the resting potassium conductance of the myenteric neurons. This effect on ion conductance is the same as that of an unknown substance that mediates slow synaptic excitations with the myenteric plexus.


The structure and function of the labrum in the lobster Homarus gammarus (L.). The labrum of decapod crustaceans is a soft lobe overhanging the mouth. The labral skeleton, musculature and innervation of Homarus gammarus are described. There are three bilateral groups of sensory neurons innervating the floor, lobe and lateral walls of the labrum. These are probably responsible for the phasic afferent activity that can be recorded from the inner labral nerve on mechanical deformation of the labrum. The labrum undergoes rhythmical retraction-protraction movements during ingestion and is shown to be active during both mandibular activity and oesophageal peristalsis. Studies were made on the duration and frequency of labral "swallowing" activity. The role of the labrum in feeding is discussed.


Oesophageal sensors and their modulatory influence on oesophageal peristalsis in the lobster, Homarus gammarus. The musculature and innervation of the oesophagus of Homarus gammarus are described as a prerequisite to studies on the mechanisms and control of food ingestion. Of particular interest are two paired sensors (the anterior and posterior oesophageal sensors) which are bilaterally situated at the oesophageal-cardiac sac valve. These are similar to contact chemoreceptors previously described in insects and are classified as such on morphological grounds and with indirect electrophysiological evidence. Oesophageal peristalsis is effected by the coordinated contraction of the Oesophageal musculature. This is controlled by rhythmical bursting neuronal activity, which can be recorded from the nerve trunks in the area. A characteristic burst recorded from the superior oesophageal nerve is used as an indication of oesophageal dilatation during peristalsis for studies on the feedback effects of the oesophageal sensors. Electrical and chemical stimulation of the posterior oesophageal sensors can initiate and increase the frequency of oesophageal peristalsis, while stimulation of the anterior oesophageal sensors can slow and terminate oesophageal peristalsis. The results are discussed and a model presented of the role of the oesophageal sensors in feeding.


Peripheral catecholamines and memory: characteristics of syrosingopine-induced amnesia. The effect of syrosingopine on retention of a passive avoidance trial in mice was investigated. The drug given in doses of 2.5, 4.0 or 6.0 mg/kg 2 hr before training, but not when given 24 or 0.5 hr or immediately after training, resulted in amnesia 7 days later. Dopamine or norepinephrine administered systemically 15 min before to 10 min after training was able to block the syrosingopine-induced amnesia. The role of peripheral catecholamines in memory formation was discussed.


Alterations in myocardial adenylate cyclase in spontaneously hypertensive rats. Particulate fractions of myocardium taken from spontaneously hypertensive rats (SHR) contained an adenylate cyclase system that was less responsive than normotensive Wistar Kyoto rats to norepinephrine, isoproterenol (mixed beta-agonist), salbutamol (beta 2-agonist), dobutamine (beta 1-agonist), dopamine, histamine, and glucagon. Addition of 5'-guanylylimidodiphosphate to the SHR myocardial preparation again yielded a lessened sensitivity to all agents except norepinephrine, dopamine, and histamine. Chronic treatment of SHR with clonidine and a high dose of propranolol produced a cardiac enzyme that was insensitive to activation by norepinephrine. Similar treatment with a low dose of propranolol did not alter myocardial responses to norepinephrine.


Long-term sleep laboratory evaluation of flunitrazepam. A sleep laboratory hypnotic drug evaluation study was conducted in which 2 mg flunitrazepam, a benzodiazepine with a half life of intermediate duration, was administered nightly to 6 insomniac subjects for 4 consecutive weeks. The drug was effective with short-term use. However, tolerance developed for sleep maintenance during the intermediate- and long-term drug administration periods. Following drug withdrawal, a significant worsening of sleep above baseline levels occurred. These data are consistent with our previous findings of rebound insomnia following withdrawal of short- and intermediate-acting benzodiazepines.


Dual histamine receptor mechanism in guinea-pig lung. Isolated guinea-pig lung parenchymal strips (GPLS) relaxed in response to the selective histamine H2-receptor agonists, dimaprit and 4-methylhistamine (4-MeH), and to low doses of histamine (10(-9) to 10(-7) mol/l) and contracted in response to several spasmogens. The order of the relative activity of the spasmogens was 2-methylhistamine (2-MeH) greater than histamine greater than carbachol greater than 2-pyridylethylamine (2-PE). Dimaprit and 4-MeH also relaxed GPLS which were contracted by 2-MeH, 2-PE or carbachol. Metiamide (a selective H2-antagonist: 5 x 10(-5) mol/l inhibited or reversed relaxations to histamine, dimaprit and 4-MeH, and significantly enhanced contractile responses to histamine without altering responses to carbachol. Mepyramine (a selective H1-receptor antagonist: 10(-8) to 10(-6) mol/l antagonized histamine-induced contractions. This investigation shows: (1) histamine is more active than carbachol in GPLS and (2) the occurrence of histamine H1-receptors mediating contraction and H2-receptors mediating relaxation in guinea-pig lung.


Studies on the inhibition of adrenaline-induced aggregation of blood platelets. alpha-Adrenoceptors of platelets seem to differ from those of other cell types. The inhibitory effect of alpha-adrenoceptor-blocking agents on adrenaline-induced aggregation of human platelets does not parallel that on adrenaline-induced contraction of rabbit aortic strips. The most potent inhibitors of adrenaline-induced platelet aggregation are the dihydrogenated ergot alkaloids of the peptide type. Among the other ergoline derivatives testes, only lisuride has a stronger inhibitory effect than those of natural ergot alkaloids.


Carbohydrate metabolism in the fetal pig during late gestation. In acute experiments on pregnant sows under sodium pentobarbitone anaesthesia, acid base balance, oxygenation and plasma metabolite concentrations were well maintained in the dam and all fetuses which remained undisturbed in utero, irrespective of the duration of the experiment. Fetal liver glycogen concentrations were also unaffected by the time of removal of the fetus. By contrast, intravascular catheterization and withdrawal of blood led to fetal hyperglycaemia and depletion of hepatic glycogen although blood gas and pH values were not changed by these procedures. In the 1 1/2--2 h sampling period following catheterization the normal positive umbilical venous-arterial differences in plasma glucose and lactate generally became reversed. These changes were prevented by the administration of hexamethonium (10--15 mg . kg-1 i.v.) but the drug did not block the fall in hepatic glycogen in catheterized fetuses. Both adrenaline and noradrenaline, which were each infused intravenously at 2.7--3.9 or 0.6--0.9 microgram . kg-1 . min-1, resulted in fetal hyperglycaemia and lacticacidemia together with a fall in arterial blood pH; hepatic glycogen concentrations in these fetuses were also reduced. The apparent sensitivity of the glycogenolytic mechanism to surgical trauma and haemorrhage in the fetal piglet is discussed in relation to findings in other species.


Comparison of substrate specificities of the human placental NAD- and NADP-linked 15-hydroxyprostaglandin dehydrogenases. A study of the relative activity of the purified placental NAD- and NADP-linked 15-hydroxyprostaglandin dehydrogenases with various prostaglandins and thromboxane B2 (TxB2) suggests that most, if not all, oxidation in the placenta of the 15-hydroxyl group of prostaglandins of the A, E, and F series as well as PGI2 (prostacyclin) and 6-keto PGF1 alpha is catalyzed by the NAD-linked enzyme. Prostaglandin B1 is an excellent substrate for the NADP-linked enzyme. Despite the conformational similarities between PGB1 and PGI2, the latter molecule is a poor substrate for the NADP-linked enzyme. Thromboxane B2 is not oxidized by the NAD-linked enzyme and is oxidized slowly by the NADP-linked enzyme.


Pharmacological comparison of isolated bronchial artery and pulmonary vein: possible functional relationships. Using selective agonists and antagonists, active alpha and beta 1-adrenoceptors and muscarinic cholinoceptors were identified in isolated strips of bronchial artery and pulmonary vein. Beta 2 receptors were located in the bronchial artery only. The bronchial artery was more sensitive to phenylephrine and less sensitive to carbachol than the corresponding pulmonary vein, which was in turn more sensitive to the spasmogenic action of serotonin, SRS-A and prostaglandins than the artery. Both vessels contained histamine H1-receptors and tryptamine D-receptors. Histamine H2-receptors were confined to the pulmonary vein. The data are consistent with the involvement of both bronchial and pulmonary vasculature in allergic pulmonary diseases.


Effect of beta-adrenergic drugs on plasma glucose and glucagon in Japanese quail: a preliminary report. The present study was conducted to determine the effects of beta-adrenergic stimulation on plasma glucose and glucagon (IRG) levels in Japanese quail. Isoproterenol, epinephrine and three relatively selective beta-adrenoceptor agonists (terbutaline, salbutamol and reproterol) produced dose-related hyperglycemia and hypoglucagonemia. This study demonstrates that beta-adrenoceptor agonists produce hyperglycemia in birds as they do in mammals, but that the rise in plasma glucose in birds, unlike mammals, is accompanied by a profound fall in plasma IRG levels.


A selective medium facilitating the isolation and recognition of Bordetella bronchiseptica in pigs. A medium was developed that in most instances allowed the isolation from pigs of Bordetella (Alcaligenes) bronchiseptica in pure, or virtually pure, culture from such sites as the nasal cavity, which contains many other bacteria. If not suppressed, some of the latter can inhibit, sometimes completely, the growth of the bordetellae on artificial media. Besides being markedly selective, the new medium is simple to prepare, reasonably cheap and practically noninhibitory to B bronchiseptica. This organism produces a distinctive colony that is easily differentiated on morphological grounds from the few other bacteria (such as Alcaligenes faecalis and Pseudomonas spp) which also grow on this medium. Parallel examination of specimens from the nasal cavities of 219 pigs in the field in southern England confirmed that this medium was better than hitherto recommended for the detection of B bronchiseptica. This was particularly true when the organism occurred in small numbers only and/or when bacteria little affected by the inhibitors in the previously recommended media were present in the nasal flora.


Effect of hypocapnia on intracellular pH during metabolic acidosis. Separate and combined effects of acute metabolic acidosis and hypocapnia were determined in skeletal and cardiac muscles of intact rats. Normocapnic metabolic acidosis, imposed by intraperitoneal injection of hydrochloric acid (6 mEq/kg), did not change skeletal muscle intracellular acid--base parameters. Hypocapnia, induced by mechanical hyperventilation, resulted in intracellular alkalosis within skeletal muscle during both respiratory alkalosis and compensated metabolic acidosis; changes of skeletal muscle intracellular bicarbonate concentration per unit change in carbon dioxide tension were identical during these two experimental procedures. These data suggest that processes other than physicochemical buffering neutralize protons taken into skeletal muscle cells during acute metabolic acidosis. The acid--base state of the heart was quite stable during these experimental manipulations; thus, it appears that cardiac muscle has an extraordinary buffering ability. Moreover, our data suggest that processes other than physicochemical buffering maintain cardiac intracellular pH normal during hypocapnia.


Free T4, free T3 and free reverse T3 in dialysates of serum. The influence of electrolytes and pH with special reference to the physiological range. The influence of electrolyte composition and pH of buffer on free T4, free rT3, and free T3 concentrations in serum dialysates has been evaluated, using an equilibrium dialysis method with subsequent direct radioimmunoassay estimations of the dialysate. Cl-, HCO3- and H+ were found to be the important ions which at concentrations similar to those occurring in vivo decreased the protein binding of all three hormones in serum, T3 being much less sensitive to these changes than T4 and rT3. The possible influence of pathological concentrations of electrolytes in serum on estimated free thyroid hormone concentrations is discussed. Because the effect of in vivo changes in serum Cl-, HCO3- and H+ on free thyroid hormone concentrations to a certain extent tend to counteract each other, it is concluded that electrolyte and pH disturbances only to a minor extent can affect measurements on free T4, free rT3 and free T3 concentrations in serum obtained by an equilibrium dialysis method.


Role of preload and inotropy in stroke volume regulation at constant heart rate. The relationship between preload and inotropy on left ventricular function was studied in anaesthetized open-chest dogs, by measuring left ventricular dimensions and stroke volume before and during saline infusion at different levels of inotropy. Left ventricular dimensions were continuously estimated by recording myocardial chord length (MCL) in the anterior wall of the left ventricle by ultrasonic technique. The effects of isoproterenol, a stimulator of adrenergic beta-receptors (high inotropy), and propranolol, an inhibitor of adrenergic beta-receptors (low inotropy), were examined during right atrial pacing at constant heart rate averaging 161 +/- 5 beats/min. Stroke volume was varied within the range 9.0 +/- 1.7 ml to 28.6 +/- 3.2 ml by increasing inotropy and preload. To increase preload, saline was infused intravenously until end-diastolic MCL increased by about 10% and left ventricular end-diastolic pressure was higher than 10 mmHg. At constant heart rate and blood volume, both before and during saline infusion, end-diastolic MCL was not influenced by isoproterenol or propranolol administration. End-systolic MCL was reduced by raising inotropy. The difference between end-diastolic and end-systolic MCL, the systolic myocardial shortening (MS), increased during saline infusion; the relative increase in MS was the same at high and low inotropy. On average, MS was more than 50% longer at high than at low inotropy, both before and after saline infusion. Thus, left ventricular end-diastolic volume is increased by saline infusion and end-systolic volume is reduced by increasing inotropy. Preload and inotropy exert independent effects on stroke volume.


Cardiac performance: optimal heart rate for maximal cardiac output. To determine optimal heart rate for the maximal cardiac output at various levels of inotropy and blood volume, the relationship between heart rate (HR) and stroke volume (SV) was examined in anaesthetized dogs during right atrial pacing. Myocardial inotropy was raised by intravenous infusion of isoproterenol, a stimulator of adrenergic beta-receptors, and reduced by propranolol, an inhibitor of adrenergic beta-receptors. Circulating blood volume was increased by saline infusion. Within the range of optimal heart rate, SV and HR were inversely related: SV = k (HR0-HR), where k indicates the relationship between changes in SV and HR. The intercept with the HR axis is HR0. At constant HR a rise in inotropy increased SV and a fall in inotropy reduced SV. These changes in SV were eual at every HR, and k was therefore constant. In contrast, blood volume expansion increased SV more at low than at high HR (k increased), but HR0 was not significantly changed. Calculated maximal cardiac output: k.HR02/4, and optimal heart/rate: HR0/2, agreed with observations when maximal cardiac output was raised from 1900 to 4500 ml/min by increasing blood volume and inotropy. Optimal HR was not influenced by changes in blood volume, but was increased from 160 to 200 beats/min by increasing inotropy. We conclude that the optimal heart rate and the maximal cardiac output can be predicted from the linear relationship between SV and HR during right atrial pacing.


Influence of beta-adrenoceptor blockade on leg blood flow and lactate release in man. Nine healthy male volunteers were studied at rest and during exercise before and after acute local beta-blockade in one leg. Oxygen uptake, arterial pressure, heart rate, leg blood flow and arterial-femoral venous differences for oxygen and lactate were determined. In addition, five subjects were studied at rest during adrenaline infusion to test the blockade; in this situation the increase in leg blood flow and decrease in resistance seen in the control leg were prevented in the blocked leg. During exercise, the beta-blockade did not influence leg blood flow. At rest, lactate release was abolished from the blocked leg, and during exercise the release was reduced by 50%. These findings demonstrate that acute betareceptor blockade does not interfere with the exercise-induced vasadilation but has metabolic consequences reflected by a reduction in the release of lactate from the leg.


Stimulation of heart rate by insulin: uninfluenced by beta-adrenergic receptor blockade in rabbits. Intravenous injection of insulin increased heart rate approximately 20% in six alloxan-diabetic rabbits. Blood glucose concentrations after insulin did not decrease below the fasting level of non-diabetic animals and none of the rabbits had signs of hypoglycermia. Intravenous injection of saline or insulin solvent had no effect on heart rate. The stimulatory effect of insulin on heart rate was not influenced by autonomic nervous blockade by propranolol or by propranolol plus atropine.


Colloidal gold probes--a further evaluation. Colloidal gold provides an immuno- and cytochemical marker suitable both for TEM and SEM. Various parameters involved in the preparation and stability of gold markers of various sizes and of probes have been reviewed and modifications in methodology suggested based on a study of factors affecting the performance of the gold marker system. In particular, dependence of protein adsorption on pH conditions correlating with protein pI is further confirmed. Presence of bioactive protein in gold probes is demonstrated and quantitated by radioassay (125I) and agglutination procedures.


[Chronic respiratory insufficiency in general practice]. The clinical entity of chronic respiratory insufficiency involves a variety of lung diseases with different etiology. Diagnosis, treatment and follow-up are mainly a problem of ambulatory medicine. The most common representative is the patient with chronic obstructive lung disease. The wide array of diagnostic tools makes the selection of appropriate tests difficult for the general practitioner. The clinical findings, as the most important aspect, together with a simple spirometric test (VC and FEV1), however, provide the physician with sufficient parameters to evaluate the current status. Blood gas analysis are indispenable, but can be performed at lengthy intervals. The main therapy relies on selective beta-agonists in combination with steroids. For long-term administration of steroids, however, inhaled steroids should be preferred. Antibiotics should be used liberally and without delay. Cardiac therapy is an important cornerstone in the treatment of chronic respiratory insufficiency. The frequency of glycoside intolerance makes diuretics the ideally suited drug for treatment of concomitant cardiac insufficiency. It is advantageous to use consistent inhalation therapy for drug administration, mainly due to the milder systemic side-effects. A choice must be made between the simple electric nebulizer or the more demanding IPPB-respirator, which requires more demanding patient selection. Both types can be leased through Cantonal institutions, which also provide for regular control of inhalation techniques and maintenance of the inhalation devices. Physiotherapy plays an important part in the instruction of patients as well as providing them with psychological assistance and support in everyday problems. Regular monthly controls by a physician with a standardized questionnaire have proven most useful in guiding the patients and adjusting therapy to individual needs. The frequency of hospitalization can only be reduced by observing all the above mentioned factors.


[Mouth and throat disinfectant preparation--dental aspects]. The oral fermentability of 10 mouth and throat disinfectant lozenges available in Switzerland were tested telemetrically. Because of their high sugar content all products produced immediate and long-lasting pH depressions in the interdental plaque except for a sugar-free preparation intended for diabetic patients. The cariogenic potential as well as the contraindication of such tablets in the treatment of infections of the oral mucosa, gingivitis, periodontitis or following dental surgery are shown.


[Causes of tooth loss in Switzerland. Results of an inquiry of private dental practitioners]. The reasons for tooth loss in different age groups were analysed. The data were compiled from questionnaires sent to 99 private dental practices chosen at random in Switzerland. Caries was the most common reason given for tooth extraction in younger age groups. After 40 years of age, however, the number of teeth extracted because of periodontal disease was higher than the number removed for any other reason.


[Neurocristopathies : embryology, physiology, and pathology of the neural crest derivatives (author's transl)]. The authors review various of the derivatives of the neural crest, consider their relationship with the APUD System, and deduce the physiopathological consequences applicable to these cells, whose only similarity lies in their common embryologic origin. They propose a system of classification of interrelated pathological conditions that these cells may show, an detail clinical information as well as the mode of genetic transmission of the principal neurocristopathic syndroms.


[Chronic lymphocytic leukaemia with cutaneous lesions and deafness following periarteritis nodosa (author's transl)]. A case is reported of a 49 years-old man who presented with cutaneous and muscular manifestations revealing periarteritis nodosa. Two years after detection of this disease occured a preleukemic condition and four years later a chronic lymphocytic leukaemia is discovered. In clinical course appeared a tuberous infiltration of the face and a sudden bilateral deafness. Regression of these two abnormalities is obtained after administration of prednisolone. The patient died of generalized infectious process postoperatively. From these clinical and pathological findings relations between periarteritis and CLL, occurence of deafness and cutaneous lesions are discussed. The relationship between the development of CLL and the immunosuppressive treatment is considered.


[Neuroleptics and analgesia (author's transl)]. Recent studies have shown that the explanation given for the analgesic effects of neuroleptics based only upon modification of pain interpretation is not valid at the present time. Studies by K. Ramabadran, J. J. C. Jacob, Greese et al., of Leysen and Smee and Overstreet, which take into the account the secretion of endogeneous "ligands" enkephalins and endorphins, offer the possibility of a better approach for understanding the role of neuroleptics in the control of pain. We have attempted to use this interpretation to explain the regular and constant efficacy of tiapride against pain generally and particularly in headaches.


The effect of etilefrine (Effortil) on central hemodynamics and aorto-coronary bypass blood flow. A pre-operative study. In an attempt to elucidate the function of an aorto-coronary bypass in patients with coronary artery obliterative disease, intravenous injection of etilefrine was used to bring about variations of central hemodynamics. Etilefrine proved to cause a significant increase of cardiac output (QPA), mean systemic blood pressure and aorto-coronary bypass flow. Calculations of peripheral myocardial resistance showed a rather pronounced decrease in all vascular regions studied. There was no change of central venous pressure. The decrease of the myocardial vascular resistance, as shown by the increase of aorto-coronary bypass flow, is indicative of the myocardial vascular capacity. Hence, it may be of value as a prognostic test.


Protein synthesis inhibition in rat liver by the mycotoxin patulin. Patulin, a carcinogenic mycotoxin, inhibits in vivo and in vitro protein synthesis in rat liver. The in vivo inhibition culminates 5 h after toxin administration and reaches a maximum of 65% with regard to control values; a breakdown of polysomes is associated with the translational blockage. However in in vitro systems, the cellular fractions obtained from patulin-treated rats appear equally as active in protein synthesizing ability and as sensitive to the toxin action as those prepared from control animals. The in vitro inhibition by patulin is dose related. The postmitochondrial system is less sensitive than that functioning with isolated polysomes and pH 5 enzyme. This difference might be due to the presence of soluble factor(s) that counteract patulin action. We propose that the inhibition of protein synthesis by patulin may result from an interaction of the drug with active SH groups at the membrane level (amino acid transport, ion equilibrium) and/or at the cytoplasmic level (enzymes and factors involved in the translational process).


Immunospecific depletion of graft-versus-host-reactive lymphocytes using sensitized syngeneic initiator T lymphocytes. We investigated a model of a lethal graft-versus-host (GVH) reaction with the aim of depleting donor spleen cells of immunospecific GVH-reactive lymphocytes. In previous studies of the recruitment of effector T lymphocytes by sensitized syngeneic initiator T lymphocytes (ITLs) we found, using a local GVH reaction, that precursors of specific GVH-reactive lymphocytes were recruited to a draining lymph node. In this study, adult F1 hybrid mice were lethally irradiated and reconstituted with 2 x 10(6) syngeneic bone marrow cells and varying numbers of spleen cells from parental strain mice. To deplete donor spleen cells of GVH-reactive lymphocytes, parental strain mice were given injections in the hind footpads 6 days earlier of syngeneic ITLs that had been sensitized in vitro against allogeneic fibroblasts. We found that injection of ITLs sensitized against the relevant allogeneic antigens led to a marked decrease in the specific GVH potential of donor spleen cells. These findings show that GVH-reactive lymphocytes can be depleted selectively by activating their recruitment to particular lymph nodes, using syngeneic ITLs.


[Changes in intercellular exchange intensity in an L cell culture]. Intercellular junctions permeable to fluorescein Na were studied with the aid of intracellular glass microelectrodes in the cultures of transformed mouse-embryo cells (L-strain). The degree of coupling was correlated with the cell culture density. In addition, the degree of coupling increased after short-time incubated of the cells with 1mM lanthanum. Such an incubation did not influence other features of cells: the value of the membrane potential, the character of growth or the inclusion of the vital dye neutral red. Some other factors stimulated cellular aggregation (concanavalin A, protamine, decreasing the pH) but did not influence the degree of coupling.


The use of ion-exchange resins as a blood preservation system. A system using ion-exchange resins for the storage of whole blood was investigated. Blood was collected into CPD, thoroughly mixed and then divided into equal amounts. To one half of the split units, four grams of phosphate charged Amberlite IR-45 resin were added, while no additions were made to the other split units. Both aliquots were stored at 4 C, adhering to routine blood banking standards. The blood stored with the resin for up to 28 days maintained high levels of 2,3-DPG and adequate levels of ATP. This new resin system has several advantages over the present blood preservation technology. There is initial increase in pH that promotes greater glucose utilization. Inorganic phosphate is provided to maintain both ATP and 2,3-DPG levels. The resin buffers the blood and maintains a narrower pH range for red blood cell enzyme activity. The resin is totally inert, completely filterable and has the ability to adsorb bacteria from the suspending solution.


Aggregation response of human platelets stored at 22 C as platelet-rich plasma. The aggregating response of human platelets stored at 22 C for 72 hours has been studied. Platelets were stored as platelet-rich plasma in order to maintain the plasma pH essentially constant. The response to ADP and collagen decreased with time, but the decrease was less with relatively high concentrations of the aggregating agents. The response to the ionophore A23187 was essentially unaltered during the storage period. Synergistic aggregation with combinations of ADP, collagen and ionophore A23187 was observed with platelets stored for 72 hours under conditions where singly each of the stimuli caused little or no effect. Stored platelets underwent reversible aggregation over a wide range of ADP concentrations but irreversible aggregation was observed in the presence of nonaggregating concentrations of collagen or ionophore A23187. The inhibition of aggregation by PGE1 or cyclic AMP was facilitated as a result of storage. It is suggested that the decreased response toward ADP and collagen reflects a reduced ability on the part of the platelets to mobilize calcium.


[Purification of Salmonella abortus ovis H antigen by using immunoadsorption]. OH-antiserum was first produced in rabbit immunization with Salmonella abortus ovis cultures and was later used for obtaining an immunoadsorbent after the method of Avrameas and Thernynck. The raw (not purified) H-antigen produced after the method of McCoy et al., was purified by the immunoadsorbent and its activity and specificity were studied serologically by the reaction of Ouchterlony. It was established that the antigen reacts with the anti-H-salmonella serum, but does not react with normal sheep serum, which is an indication that it is purified and active. It was established also that immunoadsorption is a method suitable for H-antigen production from bacterial cilia.


Transport of radiolabelled glycoprotein to cell surface and lysosome-like bodies of absorptive cells in clutured small-intestinal tissue from normal subjects and patients with a lysosomal storage disease. The transport of 3H-fucose- and 3H-glucosamine-labelled glycoproteins in the absorptive cells of cultured human small-intestinal tissue was investigated with light- and electron-microscopical autoradiography. The findings showed that these glycoproteins were completed in the Golgi apparatus and transported in small vesicular structures to the apical cytoplasm of these cells. Since this material arrived in the cell coat on the microvilli and in the lysosome-like bodies simultaneously, a crinophagic function of these organelles in the regulation of the transport or secretion of cell-coat material was supported. In the absorptive cells of patients with fucosidosis or Hunter's type of lysosomal storage disease, a smiliar transport of cell-coat material to the lysosome-like bodies and a congenital defect of a lysosomal hydrolase normally involved in the degradation of cell-coat material, can explain the accumulation of this material in the dense bodies.


Morphometric analysis of skeletal response to vitamin D in uremic rats fed a low calcium diet. The response of tibial metaphyses to pharmacologic levels of vitamin D in uremic rats fed a low calcium diet was evaluated morphometrically. Uremic (5/6 nephrectomized) rats given vitamin D had increased percent metaphyseal hard tissue, trabecular surface perimeter and percent trabecular osteoid surface and reduced numbers of osteoblasts and osteoclasts per millimeter of trabecular perimeter compared to either uremic rats given placebo or sham-operated rats given vitamin D. It was concluded that the resistance of metaphyseal trabeculae in uremic rats to vitamin D was due in part to the increase in osteoid-covered surfaces which inhibited osteoclasis and subsequent remodeling. The pathogenesis of worsening osteomalacia as a consequence of vitamin D administration to uremic rats on a low calcium diet remains unclear.


Formaldehyde-fluorescamine-induced fluorescence of mammary carcinoma cells. Lack of concordance with occurrence of oestrogen-binding receptor proteins. The combined formaldehyde-fluorescamine technique demonstrates fluorescence by mammary carcinoma cells whereas cells of the normal gland or of benign tumours do not show fluorescence. We have studied the correlation between histological staging of the disease, concentrations of oestrogen-binding proteins and the occurrence of formaldehyde-fluorescamine (FF)-inducible fluorescence. Our results demonstrate that the FF-technique detects all types of mammary carcinoma cells irrespective of their concentration of oestrogen receptors. Hence, the FF-technique represents a valuable tool for detecting both hormone-responsive and hormone-unresponsive malignant cells of the mammary gland.


Tight junctions adjacent to tumor stromal interface in human invasive transitional cell carcinomas. Tight junctions adjacent to the tumor stromal interface in invading neoplastic cells of human urinary bladder carcinomas were observed. Basal lamina, collagen and elastic fibers, and cellular debris were found next to the tight junctions. An association between microenvironment (i.e., tumor necrosis) of the invading neoplastic cells and tight junction locations was suggested.


Consequences of recurrent phosphate trapping induced by repeated injections of 2-deoxy-D-galactose. Biochemical and morphological studies in rats. 2-Deoxy-D-galactose, in a dose of 3 mmol/kg, was administered intraperitoneally twice daily to young rats for periods up to 12 weeks. This dosage schedule resulted in recurrent phosphate trapping predominantly in liver. UTP deficiency was excluded by simultaneous uridine injections. Phosphate trapping was caused by the rapid accumulation of 2-deoxy-D-galactose 1-phosphate and was most pronounced in liver but also demonstrated in small intestine, brain, spleen, and thymus. The marked, although transient, drop in the hepatic content of inorganic phosphate triggered the catabolism of adenine nucleotides and a loss of ATP. Other metabolic pathways affected by phosphate deficiency include glycogenolysis and glycolysis. Increasing with time, repeated doses of the galactose analog led to retardation and arrest of growth, hepatomegaly, and splenomegaly. The average relative liver and spleen weights were elevated 2.5- and 4.5-fold, respectively, after 12 weeks of treatment. Liver damage was indicated by hyperbilirubinaemia and a progressive rise in the activity in plasma of sorbitol dehydrogenase, alkaline phosphatase, and gamma-glutamyltransferase. Examination by light and electron microscopy showed increasing numbers of vacuoles, surrounded by a single membrane, in hepatocytes, sinusoidal endothelial cells, and Kupffer cells. Focal cytoplasmic degeneration in hepatocytes was occasionally indicated by formation of autophagic vacuoles and finger print lysosomes. Hepatocytes of 2-deoxy-D-galactose-treated rats showed a dissociation and fragmentation of the rough endoplasmic reticulum. Sinusoidal endothelial cells and Kupffer cells were markedly enlarged, the latter contained a PAS-positive but amylase resistant substance. Extrahepatic changes included an increased occurrence of vacuolated cells in thymus. Phosphate trapping and its metabolic consequences are common phenomena in the experimental injury induced b 2-deoxy-D-galactose and in some hereditary diseases such as uridylyltransferase deficiency galactosaemia, fructose intolerance and glucose-6-phosphatase deficiency.


Inhibited autophagic degradation of cytoplasm during compensatory growth of liver cells after partial hepatectomy. The livers from 56 sham-operated and 56 partially hepatectomized male albino rats killed 4--81 h after operation were investigated by electron microscopic morphometry. Following partial hepatectomy, the principal changes in volume fractions in the hepatocellular cytoplasm were: decrease of glycogen and, to a lesser extent, of mitochondria together with considerable increase of fat droplets. The volume fraction of microbodies (= peroxisomes) showed no significant difference between control and regenerating liver. By evaluating large test fields of about 40,000 micrometers 2 sectioned cytoplasm per animal it could be demonstrated that the volume fraction and the numerical density of autophagic vacuoles (AV's) were significantly reduced after partial hepatectomy. The extent of this reduction depended on the postoperative time interval. AV's were reduced by 75% at day 0 (4--17 h p.o.), by 98% at day I (19--33 h p-o.), by 75% at day II (43--57 h p.o.), and still by 50% at day III (67--81 h p.o.). The different types of AV's, defined on the basis of the different cytoplasmic components enclosed, were reduced to similar extent during the respective time periods. The reduction of AV's seems to be specific for the regenerating organ since no significant differences in the volume fraction of AV's could be found in the proximal tubular cells of the kidney of partially hepatectomized animals when compared with those of sham-operated controls. The inhibition of intracellular autophagic degradation in regenerating liver has its biochemical equivalent, i.e. inhibited protein catabolism, and is interpreted as an important and adequate mechanism in effecting the shift from the physiological steady state between anabolism and catabolism to the positive balance which is required for the compensatory growth of the liver after partial hepatectomy.


Cell population kinetics of 1,2-dimethylhydrazine-induced colonic neoplasms and their adjacent colonic mucosa in the mouse. The parameters of cell population kinetics of symmetrical 1,2-dimethylhydrazine-induced colonic neoplasms and their adjacent colonic mucosa in the mouse were analyzed using the fraction labeled-mitoses curve method and compared with those of three groups of epithelial cells in the crypt of the descending colon of normal mouse. The analysis of three groups of epithelial cells in the crypt of normal mouse indicates that differentiation of epithelial cells was associated not only with a smaller proliferative pool of cells but also with a shortening of the duration of G2 phase and a prolongation of mitotic time. Other parameters of cell cycle did not change significantly. The mean cell cycle time of neoplastic cells in chemically induced colonic neoplasms was similar to that of epithelial cells in normal colon, but the variance was much greater in neoplastic cells. In neoplastic cells, the proliferative pool was greater, the G1 phase prlonged, and the S phase and the mitotic time became shorter as compared to epithelial cells in normal colon. The duration of G2 phase of neoplastic cells fell between the values of presumptive stem cells and differentiating cells in normal colon, compatible with the hypothesis that neoplastic cells are transformed stem cells defective in cellular differentiation. In the colonic mucosa immediately adjacent to neoplasms, the fraction-labeled-mitoses curve showed a flat second wave, indicating that the group of cells initially labeled by the pulse became a mixture of cells, some continuing the proliferative cycle normally, some going out of cycle, some slowing down in their passage from S through G2 to M, and some being arrested in mitotic phase. Such heterogeneous behavior of cells may be closely related to expansion of neoplasms. With some assumptions, however, cell cycle parameters of those normally cycling cells were estimated: the cell cycle time and the duration of G1 phase and mitotic phase were prolonged as compared to neoplastic cells and epithelial cells of normal colon.


Nitrosomethylurea (MNU): a powerful carcinogen for mouse skin. A study utilising the tetrazolium test (TZT) and topical skin applications. Nitrosomethylurea (MNU) was tested with the tetrazolium test for skin carcinogens with positive results. 39 hairless mice were painted once a week with a 1% solution of MNU in acetone. After 18 weeks all the animals bore tumours and 90% had skin carcinomas. Hence, the strong carcinogenic potency of MNU was confirmed, and the validity of the tetrazolium test was given further support.


Diffuse glomerulosclerosis--a dysfunction of the mesangium? A morphometric study of the rat's kidney. The phagocytic capacity of the glomerular mesangial cells in 42 rats was inhibited by the prolonged injection of PVA. Morphometric evaluation of electron microscopic sections showed that this treatment leads to an accelerated thickening of the lamina densa of the glomerular basement membrane and of the mesangial matrix (early glomerulosclerosis). This suggests that a disturbance of lysosomal degradation can lead to glomerulosclerosis, and that the possibility of mesangial dysfunction should always be considered as a possible factor in the pathogenesis of all types of diffuse glomerulosclerosis.


Nodular paragranuloma and progressively transformed germinal centers. Ultrastructural and immunohistologic findings. Ultrastructural and immunohistologic findings in a nodular variant of Hodgkin's disease with lymphocytic predominance, called nodular paragranuloma, are presented and compared with those in so-called progressively transformed germinal centers. These are large follicles with numerous lymphocytes which can be found not only in nonspecific lymphadenitis, but also in lymph nodes from patients with nodular paragranuloma. The immunoperoxidase technique was applied on paraffin sections to detect intracytoplasmic immunoglobulin and lysozyme. The so-called L & H type Sternberg-Reed cells contained IgG and one type of light chain per cell, suggesting that such cells produce immunoglobulin. The ultrastructure of the L & H type Sternberg-Reed cells favored the immunoblastic nature of these cells. It is concluded that nodular paragranuloma differs from other types of Hodgkin's disease by its localization in B-cell areas and the presence of atypical B immunoblasts.


Stereological study of B-cell mitochondria in alloxan-treated mice. Using stereological techniques, including semi-automatic image analysis, the B-cell mitochondria were studied in the pancreatic islets from one group of control mice and two groups of mice killed 10 min and 60 min, respectively, after alloxan administration. Ten min following alloxan the mitochondrial volume and envelope surface densities, the mean mitochondrial volume and surface area, and the area of mitochondrial profiles were significantly increased, whereas the mitochondrial numerical density was not significantly altered. At the 60 min observation time the mitochondrial volume density, the mean mitochondrial volume and surface areas, and the area of mitochondrial profiles were significantly decreased, whereas the mitochondrial envelope surface was not significantly altered. The findings indicate a rapid swelling, followed by disintegration of the mitochondria in the B-cells of alloxan-treated mice, thereby supporting our view that mitochondrial lesions play a primary role in the development of alloxan diabetes. These lesions are believed to be due to ionic alterations in the B-cells ("Pi-pH hypothesis").


Stereological study of endoplasmic reticulum, Golgi complex and secretory granules in the B-cells of normal and alloxan-treated mice. Different B-cell organelles (lamellar and vesicular endoplasmic reticulum, Golgi complex, whole secretory granules and secretory granule cores) were studied stereologically in pancreatic islets from control mice and mice killed 10 or 60 min following alloxan injection. Ten min following alloxan a significant decrease was observed in the volume, surface and numerical densities of whole secretory granules and their cores, and a significant increase was found in the volume and surface densities of vesicular endoplasmic reticulum. At the 60 min observation time, a significant decrease was seen in the volume density of lamellar endoplasmic reticulum and Golgi complex, and in the volume, surface and numerical densities of whole secretory granules and their cores, and a significant increase was observed in the volume and surface densities of vesicular endoplasmic reticulum, and in the mean values for volume and surface of whole secretory granules and their cores. The stereological data indicate swelling of endoplasmic reticulum, decreased Golgi area, and decreased number and total volume and surface of secretory granules during  the first hour after alloxan administration to mice. The observations may be consistent with inhibited insulin synthesis.


Aspects of normal and dystrophic chicken muscle grown in vitro. Pectoral muscle from normal and dystrophic New Hampshire chicken embryos was dissociated and grown in vitro. Marked differences between the two types of cell cultures were observed with the light and electron microscopes during early myogenic stages. The diseased myoblasts assumed a polarized affect and fused into smaller and fewer myotubes. Pseudostraps rather than true muscle straps were often seen in diseased cultures. There was also a delay in the appearance of myosin containing thick myofilaments in differentiating dystrophic muscle cells.


Effects of actinomycin D on ribosomal RNA and protein synthesis as revealed by high resolution autoradiography. Incorporation of tritiated amino acids and uridine was studied in untreated and actinomycin D treated HeLa cells by high resolution autoradiography. Results showed a non-selective inhibition of protein synthesis by actinomycin, as measured by the decrease in radioactive amino acid uptake. When cells pretreated with actinomycin D were incubated with radioactive amino acids and uridine, amino acid uptake in the nucleolus still occurred, while uridine uptake was almost completely eliminated. These findings suggest that in the absence of ribosomal RNA precursor synthesis, nucleolar protein synthesis continues to some extent, and that this protein is transported to the nucleolus.


The distribution and mobility of surface anionic groups of normal human oesophageal epithelium following interaction with cationized verritin. Oesophageal epithelial cells from biopsies from normal patients showed the presence of randomly distributed anionic groups, mostly sialic acid on the cell membrane in fixed material shown by cationized ferritin. When biopsies were pulse labelled, patching occurred in all three cell layers. Patching was energy dependent and did not occur at 4 degrees C. Pulse labelled material incubated on an unlabelled medium showed progressive loss of cationized ferritin from the cell membrane. This was mostly into the medium, although some was internalized in membrane profiles. A second pulse of cationized ferritin produced further patching suggesting regeneration of cell membrane. Superficial cells were leaky, but their organelles were not.


[Role of carbohydrates in the manifestation of the hemagglutinating and neuraminidase activities of the influenza virus]. Suspensions of influenza virions (MRC-11, X-7, and A/PR8/34 strains) were treated with a mixture of glycosidases including alpha-D-mannosidase, alpha-L-fucosidase, beta-D-galactosidase, and beta-D-N--acetylglucosaminidase for the study of the influence of carbohydrate part of influenza virus surface glycoproteins on the hemagglutinating and neuraminidase activity. The availability of oligosaccharide chains for the effect of glycosidases was found to change in relation to the functional status of the glycoprotein molecule. When carbohydrate residues were cleaved from neuraminidase in virus particle, its enzymatic activity did not change, whereas the hemagglutinating activity of virions after treatment with glycosidases decreased more than 50-fold.


The purinergic nerve hypothesis and insulin secretion. In adrenalectomized rats the effect of i.v. injection of glucose and ATP on insulin changes in external jugular vein was determined in normal and alloxan diabetic animals. In another set of experiments the direct effect of ATP on insulin secretion was investigated. Glucose and ATP were injected in the carotid artery and the blood samples were withdrawn from the portal vein. In these experiments there was immediate and excessive production of insulin release in the portal vein after ATP injection in the carotid artery. In alloxan diabetic rats, despite the high blood glucose levels, the plasma insulin was low and did not respond to glucose stimulation. ATP could increase the sensitivity of the diabetic rats to glucose. The possible role of purinergic nerves in insulin secretion is discussed. It is concluded that multiple innervation of the islets by purinergic, cholinergic and adrenergic nerves, regulate insulin secretion. It is suggested that: 1. Purinergic nerve stimulation is more specific for insulin secretion. 2. ATP is considered the principal transmitter released from purinergic nerves causing insulin secretion. 3. The insulin stimulatory effect normally produced by glucose is through purinergic nerves. 4. It could be possible that one of the causes of diabetes is a defect in the purinergic innervation of the islet cells thus the sensitivity of the islets to glucose is decreased.


[Effect of psychotropic drugs on somatosensory evoked cortical potentials]. The effects of some typical clinically tested psychotropic drugs were studied in acute experiments on awake rats. Potentials in somatosensory cortex were evoked by peripheral stimulation. Haloperidol, diazepam, medazepam, desipramin and a new substance, the MPP-sulton, caused significant modification in latency and amplitude of some components of the potentials. Between the effects of psychotropic drugs exist differences which, in turn, were clearly distinguishable from the modification of these evoked responses by unspecific narcotic effects of hexobarbital.


[Simple method for the preparation of haptoglobin]. The present paper deals with a method permitting the isolation of haptoglobin 2-2 from human serum or plasma. The haptoglobin is adsorbed to DEAE-cellulose at pH 5.1 by batching. The loaded cellulose is given into a column and the haptoglobin eluted by a 0.1 M to 0.15 M acetate buffer. The last step is a gel-chromatography on Sephadex G-200. Disc- and immunoelectrophoresis were used to test the purity. As by-product acid alpha1-glycoprotein can be obtained.


Effect of a deproteinized blood extract on experimental granulation tissue. The present work was undertaken to study the effects of a deproteinized extract of calves' blood (Solcoseryl) on developing granulation tissue in rats. Cylindrical hollow viscose cellulose sponge implants were used as an inductive matrix for the growth of granulation tissue. In the first, sham group the implants were treated daily by withdrawing 1 ml of wound fluid from the central dead space of the implant and then re-injecting the fluid. In the second, experimental group the aspirated wound fluid was replaced by a corresponding volume or Solcoseryl. Analyses of wound fluid and granulation tissue were carried out 4, 10 and 21 days after implantation. A statistically significant increase of granulation tissue hemoglobin (+21%) was observed at 10 days in the Solcoseryl group as compared with the sham-treated rats, indicating an enhanced capillary ingrowth. Concurrently, the mean amount of DNA in the Solcoseryl-treated tissues was elevated by 48% over the level of the sham-treated group, demonstrating an augmented cellularity of granulation tissue. At 21 days the mean amount of collagen hydroxyproline of the Solcoseryl group was 31% above the level measured in the sham-treated animals. PO2, PCO2 and pH in the wound fluid and the amounts of RNA and uronic acids showed no essential differences between the two groups. These data demonstrate a stimulatory effect of Solcoseryl on several aspects of granulation tissue formation: augmented vascularization, elevated cellularity and subsequent enhancement in the accumulation of collagen.


Bone marrow transplantation for severe aplastic anemia. A report of 9 cases. 9 patients with severe aplastic anemia (SAA) were treated with bone marrow transplantation (BMT). 5 were conditioned with cyclophosphamide and received and HLA-identical graft (4 patients) or a mismatched graft (1 patient): 1 rejected the graft on day 30 and died on day 34 during conditioning for a second transplant; 1 died on day 15 with acute and severe graft versus host disease (GvHD) in the absence of haemopoietic engraftment; 3 are alive and complete chimeras at 1,069, 490 and 332 days after transplantation. GvHD developed in 4 patients and was treated successfully in 3 with high dose methylprednisolone and/or antilymphocytic globulin (ALG). 4 patients were conditioned with ALG and received bone marrow from a haploidentical sibling or parent: 1 patient was refractory; 3 patients showed evidence of hematologic reconstitution, but 2 of these required a second course of ALG. 3 patients in this group are alive between 60 and 490 days; 1 patient died on day 121 of HBSAg-negative acute hepatitis.


Relationship between the dipole moment and rate of absorption of drugs. Ten drugs were measured for the rate of buccal absorption and examined for the relationships between the rate constant for absorption and molecular physicochemical parameters. On the ground of the results the molecular dipole moment seems to provide more reliable information on the rate constant of buccal absorption than does the water-lipid partition coefficient or the diffusion constant. Additional perfusion studies on isolated gut were also performed in some cases. A mathematical model was developed for the calculation of the rate constants for those cases in which pH of the system changes linearly with the logarithm of time.


Blood-brain barrier to albumin in awake rats in acute hypertension induced by adrenaline, noradrenaline or angiotensin. Acute hypertension was induced by adrenaline, noradrenaline or angiotensin in awake unrestrained rats with chronic indwelling catheters in a jugular vein and in the aorta. The leakage of 125IHSA (human serum albumin) into the brains from rats given adrenaline was significantly larger than in the brains from rats given noradrenaline or angiotensin. It is likely that the enhanced vulnerability of the blood-brain barrier to an adrenaline-induced increase in blood pressure is due to the beta-adrenergic stimulating effect of adrenaline.


Neonatal 6-hydroxydopamine treatment increases the vulnerability of the blood-brain barrier to acute hypertension in conscious rats. Acute hypertension increases the cerebrovascular permeability to protein to a higher extent in anesthetized than in conscious rats. When hypertension is combined with a pronounced cerebral vasodilatation, e.g. in bicuculline-induced seizures, the protein leakage is enhanced. Conscoius, unrestrained 2--3-months-old rats received adrenaline or bicuculline i.v. during continuous recording of the mean arterial pressure and were killed 3 minutes later. Rats, neonatally sympathectomized by 6-hydroxydopamine, had significantly increased extravasation of 125I serum albumin in the brain after adrenaline-induced hypertension than nonsympathectomized rats. Since transection of the cervical sympathettic trunk alone does not have the same effect, a protection of the blood-brain barrier in acute hypertension in conscious rats may, at least in part, be mediated via the central noradrenergic innervation of cerebral vessels. Bicuculline did not increase blood pressure in 6-OHDA treated rats; thus the blood-brain barrier remained intact.


The calcium selective electrode--a possible tool in dental research. Ion selective potentiometry is widely used for analytical purposes in biologic research. The Ca selective electrode offers an inexpensive and easily handled means for studying ionized calcium. It does not affect the process studied, and may, for example, combined with an automatic recorder, give running information on dynamic systems. The calcium electrode responds in accordance with the Nernst equation and has a theoretical range of 1--10(-6) M Ca2+. The electrode is influenced by changes in temperature, in pH and also in concentration of buffer solution. The Ca sensitive membrane is adversely affected by acetic acid but tolerates lactate/lactic acid buffers. Best linearity in calibration curves for Ca2+ in lactate buffers is found in concentrations from 10(-4) M to 10(-2) M Ca. In a 5% Na lactate/lactic acid buffer, pH 5, with 0.5 M KNO3 as an ionic strength adjuster, a straight calibration curve, Nernstian slope factor 27 mV/decade, is found from 10(-4) to 1 M Ca.


The hydrolysis rates of ATP and phosphoserine in the odontoblast-predentin region of intact and carious human teeth. Coronal odontoblast-predentin tissue was taken from intact teeth and from teeth with carious lesions of varying depths, and the samples (20, 200 x g, saline-soluble fractions) were studied for their ability to hydrolyze adenosine triphosphate (ATP) and phosphoserine. A significantly higher rate of ATP hydrolysis at pH 9.8 was detected in enamel caries and a significantly lower rate in advanced caries than in intact teeth. The rates of the hydrolysis of phosphoserine at pH 9.8 did not differ between the various tooth groups, and the only clear trend indicating elevated enzyme activity was seen in the group of initial dentin caries. The ATP and phosphoserine hydrolysis at pH 9.8 were considered to have been due to the nonspecific alkaline phosphatase (EC 3.1.3.1) activity. In the presence of both levamisole and ouabain, maximal enzyme-dependent ATP hydrolysis was observed at pH 7.9. The remaining ATP-cleaving activity was thought to have been due to the Ca2+Mg2+-dependent ATPase (EC 3.6.1.3). The Ca2+Mg2+-dependent ATPase activities at pH 7.9 remained at constant levels in both the intact and carious groups. The parameters studied in vitro were thought to reflect changes in the mineralization rate of reparative dentin as a response to caries in vivo.


Effects on fetal breathing movements of maternal challenges. Cross-over study on dynamic work, static work, passive movements, hyperventilation and hyperoxygenation. Ten women in the last trimester of a normal pregnancy were subjected to five different loads in a cross-over study. Fetal breathing movements (FBM), fetal heart rate (FHR), maternal heart rate (MHR), and mean arterial pressure (MAP), maternal transcutaneously measured pO2 (Tc-pO2), and the energy supply to the Tc-pO2 electrode were recorded continuously before, during, and after the load. Maternal capillary pH and pCO2 were measured at three representative time points. The immediate responses of the incidence of FBM to the different challenges were: increase after dynamic work (bicycle test); no change after static work (isometric muscle contraction) and passive movements; decrease after hyperventilation and hyperoxygenation. FHR was unaffected by all challenges. The FBM incidence varied in parallel with pCO2 after dynamic work and hyperventilation and inversely with the Tc-pO2 rise caused by hyperoxygenation. Maternal pH was increased after passive movements (no change in FBM) and after hyperventilation (decreased incidence of FBM), FBM seem to be more sensitive to environmental changes than is the FHR. Mechanical stimuli to the uterus were not responsible for the augmentation of FMB seen after the bicycle test. The present observations reveal the multifactorial nature of the regulation of FBM, and support the role of CO2 as a major stimulator of breathing movements also in prenatal life.


Relationships between oxygen and carbon dioxide tensions and acid-base balance in arterial blood and in medullary blood from long bones in dogs. By means of an invasive technique the relations between arterial and medullary gas tensions PO2 and PCO2), and arterial and medullary acid-base balance (pH and standard bicarbonate) were determined in long bones in seven anaesthetized dogs. A semilogarithmic correlation was found between the arterial oxygen tension and the oxygen tension in the medullary blood. Between the arterial carbon dioxide tension and the medullary blood carbon dioxide tension a linear correlation was demonstrated A linear correlation was also found between arterial pH and standard bicarbonate values and the corresponding values obtained from medullary blood. With regard to the parameters investigated no difference was demonstrated between epiphyseal, metaphyseal or diaphyseal medullary blood.


Nutritionally induced necrotizing glomerulonephritis and polyarteritis nodosa in pigs. A florid necrotizing glomerulonephritis was found in all 48 pigs that were fed a waste product from the industrial production of the proteolytic enzyme Alcalase NOVO. In addition, three of the animals developed a lesion identical to polyarteritis nodosa. Focal necrosis of the glomeruli was observed in all animals. Electron microscopy showed electron dense deposits at the subendothelial and subepithelial side of the basement membrane of the glomerular capillary wall and in the mesangium. Immunofluorescence microscopy showed IgM in a fine granular pattern in the glomeruli of all 48 pigs. This appears to be the first report on nutritionally induced glomerulonephritis and polyarteritis nodosa in pigs.


Effects of digoxin on isolated human peripheral arteries and veins. In isolated human crural arteries and veins, digoxin induced slowly developing, long-lasting contractions. These contractions were not diminished by alpha-adrenoceptor blockade or by washing, but were abolished by the calcium antagonist nifedipine. In the presence of digoxin, the maximum contractile responses to noradrenaline (18 microM) and potassium (127 mM) markedly increased, and the glycoside shifted the noradrenaline concentration-response curve to the left. Immersion of vein preparations in calcium-free medium for 30 min. abolished the digoxin contraction, whereas responses could still be elicited by potassium and noradrenaline. A change of the extracellular potassium concentration from 4.6 to 6.9 and 9.2 mM caused relaxation, and a further increase to 13.8 mM contracted the preparations. After pretreatment with digoxin (1 micronM), a potassium change from 4.6 to 1.15 mM caused relaxation and all concentrations exceeding 4.6 mM produced contraction. It is concluded that digoxin has a direct contractile effect on isolated human crural vessels, and that this effect is dependent on the extracellular calcium concentration. In the presence of the glycoside, the responses to noradrenaline and potassium are potentiated. Vascular responses to changes in extracellular potassium concentration are influenced by digoxin.


Salivary secretion evoked by H80/62, (+/-)-1-(4-hydroxyphenoxy)-3-isopropylamino-2-propranol, a new beta 1-selective adrenoceptor agonist. H80/62, recently introduced as a beta 1-selective adrenoceptor agonist, caused a long-lasting flow of saliva from parotid and submaxillary glands of the rat. The effect of H80/62 was prevented by propranolol or a beta 1-selective blocker. The drug did not exert its effect via the sympathetic postganglionic nerves; and it was not taken up by the neuronal amine pump. Sympathetically decentralized or denervated glands acquired a supersensitivity of the postjunctional type to H80/62.


Slow-reacting substance of anaphylaxis:  studies on purification and characterisation. We have demonstrated the role of arachidonic acid and lipoxygenase in the genesis of SRS-A and the possible role of lipoxygenases in its metabolism. SRS-A has been purified in microgram quantities together with closely related material. We have identified 5,12 di-hydroxy-6, 8, 10, 14 eicosatetranenoic acid released by antigen challenge, and which we feel is related to SRS-A. Further structural work is in progress.


Schistosomiasis--cancer: etiological considerations. A review. Evidence for a causal connection between Schistosoma haematobium-infection and carcinoma of the urinary bladder is discussed. A group relationship of schistosomiasis cancer to cancers as+sociated with asbestosis, foreign body implants, and cicatrization is suggested on the basis of several criteria. Results of experimental foreign body tmuorigenesis in mice are presented and elaborated in relationship to schistosomiasis cancer. Carcinogenic development at the cellular level is discussed with emphasis on the essential role of tissue-environmental conditions, especially fibrotic changes and macrophage quiescence.


The endosymbionts of Glossina morsitans and G. palpalis: cultivation experiments and some physiological properties. Pyruvate, malate, and succinate are the main substrates for bacteroid respiration; oxygen uptake can be inhibited by rotenone and antimycin A, but not by cyanide. The symbionts displayed limited growth and survival for over 80 days in a medium with succinate and pyruvate as main substrates, and supplemented with nucleotides. It was not possible to cultivate the endosymbionts of G. morsitans and G. palpalis intracellularly in cell cultures of the tsetse fly or of vertebrates. A high attraction between cells and symbionts was observed in these systems; about 10% of all bacteroids were incorporatedby the cells but they were lysed and digested within 48 h.