Beta-adrenergic receptors, cyclic AMP, and ion transport in the avian erythrocyte. (1) Interaction of beta-adrenergic catecholamines with specific receptors on the cell membrane causes activation of adenylate cyclase, generation of intracellular cyclic 3',5'-AMP, and enhanced sodium and potassium transport. The effects of catecholamines on each of these functions are characteristic of beta1-type receptor interactions and are specific for 1(--)isomers. The order of effectiveness of agonists is isoproterenol greater than norepinephrine approximately equal to epinephrine. (2) exogenous cyclic 3',5'-AMP enhances sodium or potassium transport in a fashion identical to that caused by catecholamines. (3) the receptor contains an ethanolamine site that recognizes the stereospecific function of the agonist. Interaction at this site can be determined by measuring specific binding of a high specific activity-radioiodinated beta-blocker analogue (125-iodohydroxybenzylpindolol). Catecholamines bind at this site with affinities that parallel their biologic effectiveness as beta-adrenergic agonist. Beta-Blockers interact at this site with affinities that parallel their potencies as specific inhibitors of biologic function. (4) the adenylate cyclase complex contains a receptor site for guanyl nucleotides that is distinct from the catalytic site for ATP. Interaction at this site can be determined by measuring binding of radioactive guanylylimidodiphosphate (GMPPNP). GMPPNP markedly enhances the adenylate cyclase response to beta-adrenergic catecholamines with a 10-fold increase in apparent affinity of isoproterenol for activation of adenylate cyclase. Concomitantly there is a shift in Km and Vmax for ATP at the catalytic site. (5) Activation of adenylate cyclase by agonist or GMPPNP alone is reversible (GMPPNP remains bound even though the system returns to basal activity). The addition of isoproterenol plus GMPPNP, however, results in a state of enzyme activity which is apparently irreversible even though, under these conditions, the beta-adrenergic catecholamine freely dissociates from its receptor site. Under these conditions also the activated enzyme is not inhibited by the specific beta-blocker propranolol. (6) There is no evidence under adenylate cyclase assay conditions for a direct interaction of guanyl nucleotides with the beta-adrenergic receptor site nor for direct interaction of the agonist with the guanyl nucleotide site.


Behavioral effects of dibutyryl cyclic 3'5'AMP and biogenic amines in rats. White male Wistar rats were injected intraventricularly with 200 mug of dibutyryl cyclic 3'5'AMP (DAMP. 30 min later part of animals was injected intraventricularly with 10 mug of dopamine (DA), 10 mug of 5-hydroxytryptamine (5-HT) or 50 mug of acetylcholine (Ach). Immediately after injection of the amines the time of walking, convulsions, immobility and number of jumps during 10 min periods of observation was measured. DAMP induced excitation of animals expressed by convulsions, jumping and short period of immobility. All studied biogenic amines inhibited convulsions and caused periodically stereotypical behavior. 5-HT and Ach increased the time of immobility in animals injected with DAMP. DA increased the number of jumping. These results indicate the interaction between DAMP and DA, 5-HT and Ach on behavioral phenomena in rats.


Thyrotropin-receptor interaction and cyclic AMP-mediated effects in thyroid cells. Thyroid cells in culture constitute a suitable system for the study of thyroid gland function and its regulation. The natural thyroid stimulator (TSH) induces the in vitro reorganization of thyroid cells into three-dimensional follicles morphologically and metabolically similar to gland follicles. In contrast, nonstimulated cells develop as a monolayer and in a concerted manner rapidly lose the enzymes involved in iodine metabolosm and the aptitude to bind TSH to specific receptor sites. The morphogenetic action of TSH and its ability to maintain the specific metabolic properties of thyroid cells in culture are mediated by cyclic AMP via new RNA and new protein synthesis. Therefore comparison of the properties of a given cell type in morphologically and metabolically different status should provide a valuable tool for studying the TSH mechanism of action. Using 125-I-labeled TSH of high purity, high specific radioactivity, and preserved biologic potency, TSH interaction with intact cells and their derived plasma membranes was studied. At both the cellular and the sub-cellular level a very good agreement was found for the values of the rate and equilibrium constants of labeled TSH binding. A single type of high-affinity low-capacity site was revealed. In contrast, in both systems dissociation of bound labeled TSH was not of single-order kinetics and showed two kinetic components with half-lives of 3 and 30 min. An excellent correlation between half-stimulation of adenylate cyclase and iodide transport mechanism activation, and the dissociation constant of TSH binding, was found, indicating that the in vitro system studied was relevant to physiologic regulation.


Ca-2+/Mg-2+-dependent cyclic nucleotide phosphodiesterase and its activator protein. (1) Ca-2+/Mg-2+-dependent cyclic nucleotide phosphodiesterase was found in the supernatant fluids of a variety of tissues, including cerebral cortex, cerebellum, kidney, liver, and heart. (2) this enzyme required Ca-2+, Mg-2+, and an activator protein (PAF) for the activity. In the presence of these ingredients the enzyme hydrolyzed cyclic GMP preferentially when incubated with a low concentration (0.4 muM) of substrate. (3) The enzyme devoid of PAF was eluted in fraction II with a molecular weight of approximately 150,000 after Sephadex G-200 gel filtration of the supernatant fluids using medium containing EGTA. PAF thus separated from the enzyme was eluted in a fraction corresponding to a molecular weight of approximately 28,000 by gel filtration. Stimulation of the activity of fraction II by Ca-2+ was completely dependent on the addition of PAF. (4) Formation of an active enzyme-PAF complex with an estimated molecular weight of 200,000 was demonstrated by gel filtration of a mixture of the enzyme and PAF in medium containing Ca-2+. It is likely that the activity of the Ca-2+/Mg-2+ in a concentration range of approximately 1 to 10 muM, as shown in the following equation: [Enzyme] inactive + PAF + Ca-2+ in equilibrium [enzyme - PAF - Ca-2+] active. More than one PAF protein may bind to one molecule of enzyme to form an active complex. Equilibrium of the above equation is probably determined mainly by the intracellular concentration of Ca-2+ in vivo. (5) The enzyme-PAF complex was more labile after heat treatment than the free form of enzyme. (6) PAF was isolated from all tissues listed in (1). The levels of PAF and of fraction II did not correspond exactly.


Differential activation and inhibition of the multiple forms of cyclic nucleotide phosphodiesterase. The brain as well as other mammalian tissues contains several different forms of cyclic nucleotide phosphodiesterase separable by polyacrylamide gel electrophoresis. Each tissue and each individual type of cell has its own distinctive pattern and ratio of these multiple forms of phosphodiesterase. The different forms have several distinguishing properties and characteristics, and their activities may be differentially regulated both acutely and chronically. The enzyme forms have different stabilities, kinetic properties, substrate specificities, and sensitivities to an endogenous activator and to several inhibitors of phosphodiesterase. The phosphodiesterase inhibitors studied not only inhibit the different forms of phosphodiesterase to different degrees but apparently do so by different mechanisms. Thus whereas theophylline, cyclic GMP, and low concentrations of papaverine inhibit the phosphodiesterases by competing with the substrate (cyclic AMP), trifluoperazine apparently inhibits phosphodiesterase by interfering with the phosphodiesterase activator. This confers a great deal of specificity to this drug, since only one form of phosphodiesterase is markedly activated by the activator. Chronically, a specific form of phosphodiesterase appears to be inducible. This induction is probably controlled by the intracellular cyclic AMP concentration. The phosphodiesterase activator also appears to be regulatable, the age of the animal being one of the factors controlling its activity. Finally, since different types of cells have different relative amounts of the phosphodiesterases and since these forms of the enzyme can be differentially inhibited by drugs, it may be possible to develop drugs which will selectively increase the cyclic AMP concentration in discrete cell types. Evidence that cyclic AMP is involved in certain disease states suggests further that by selectively altering the concentration of cyclic AMP in these cells, one might be able to alter the course of the disease.


Mechanisms of control for cAMP-dependent protein kinase from skeletal muscle. A number of properties of homogeneous cyclic 3',5'-AMP (cAMP)-dependent protein kinase from rabbit skeletal muscle were determined. The enzyme is shown to be a tetramer consisting of one regulatory subunit dimer and two catalytic subunit monomers. Skeletal muscle protein kinase interacts with cAMP and MgATP as illustrated in the following equilibrium expression: R2C2 - (MgATP)2 + 2 cAMP in equilibrium R2 - (cAMP)2 + 2C + 2 MgATP. MgATP is shown to decrease the affinity of the enzyme for cAMP and to be necessary for recombination of the subunits. The concentration of the enzyme in tissue relative to that of cAMP is high enough to influence kinetic parameters of the activation process by cAMP. The cumulative effects of MgATP and high enzyme concentration are to increase the apparent activation constant for cAMP so that in vivo the enzyme would not be highly activated under basal conditions but would be greatly stimulated by increases in cAMP concentration. As a result, it is not necessary to invoke the concept of compartmentalization of cAMP to explain how it could regulate protein kinase activity in vivo. Finally, data are presented which indicate that a possible function of the heat-stable protein inhibitor of cAMP-dependent protein kinases may be to suppress the activity of protein kinase due to basal concentrations of cAMP. As such, the inhibitor could indirectly change the amount of cAMP needed to allow expression of protein kinase activity.


Hormonal regulation of adenosine 3',5'-monophosphate-dependent protein kinase. There appear to be two classes of protein kinases in rat heart and adipose tissue, types I and II. Type I elutes from DEAE-cellulose at smaller than 0.1 M NaCl and type II at greater than 0.1 M NaCl. The type I enzyme is more readily dissociated by salt or histone than is the type II enzyme. If the type I kinase is first dissociated by cAMP, the subunits reassociate very slowly at 0 degrees C on removal of the cAMP by Sephadex G-25 chromatography, whereas those of type II reassociate very rapidly. Rat heart contains mostly type I and a small amount of type II enzyme, whereas adipose tissue contains almost exclusively the type II enzyme. The adipose tissue enzyme resembles the heart type II kinase in all of the above properties, although the two enzymes are not identical as indicated by slight differences in elution patterns from DEAE-cellulose columns. Incubation of rat epididymal adipose tissue with low concentrations of epinephrine (0.11 muM) increases glycerol production and the fraction of the protein kinase in the active form (activity ratio). The change in cAMP under these conditions is not statistically significant. The presence of insulin inhibits the epinephrine effect on glycerol production and protein kinase but has no measurable effect on cAMP levels. Incubation of adipose tissue with high epinephrine concentrations (11 muM) increases the cAMP level, the protein kinase activity ratio, and glycerol production. Under these conditions insulin decreases the cAMP level and kinase activity ratio but does not reduce glycerol production. The data suggest that very small changes in the tissue cAMP level, undetectable by the assay method, are magnified during the stepwise activation of glycerol output aided possibly by cooperative effects between cAMP and protein kinase. The procedure developed for determining the state of activation of the cAMP-dependent protein kinase in adipose tissue must be modified by reducing the salt concentration of the buffers in order to carry out similar studies in the heart. This reflects the different types of protein kinase in the two tissues. The addition of charcoal to crude extracts of heart prevents protein kinase activation by added cyclic AMP. Charcoal should therefore prevent any activation that could occur if any sequestered cAMP were released during homogenization. Charcoal addition thereby provides a means to distinguish intracellular cAMP activation of the kinase from that which might occur following cell rupture. If epinephrine-perfused hearts are homogenized in the presence of charcoal, epinephrine stimulation of the protein kinase is only slightly decreased. This indicates that the protein kinase is activated intracellularly by cAMP and suggests that all of the cAMP in the cell is available to the protein kinase; i.e., cAMP is not released during homogenization.


Regulation of cyclic GMP levels in the ductus deferens of the rat. The cyclic GMP level in the ductus deferens is elevated by acetylcholine, norepinephrine, KCl, and the phosphodiesterase inhibitor SC-2964. The presence of extracellular Ca++ is required for the effects of all of these agents on cyclic GMP levels. In addition, Ca++ appears to be an important factor for the basal turnover of cyclic GMP in this tissue, but it may be less important in other tissues. These observations have led us to the following working hypothesis (Fig. 5): The interactions of some hormones or neurotransmitters with membrane receptors secondarily increase cyclic GMP formation after primarily increasing the influx of extracellular Ca++ or changing the distribution of Ca++ among intracellular pools or compartments. However, in addition to this possibility, other hormonal effects on particulate and/or soluble guanylate cyclase that do not involve Ca++ mediation must also be considered. Some agents that are known to increase cyclic GMP in tissues have been reported in preliminary communications to activate cell-free preparations of guanylate cyclase (Amer and McKINNEY, 1973; White, Ignarro, and George, 1973), but these reports have not yet been confirmed by other laboratories. Secretin has been reported to stimulate guanylate cyclase activity from several tissues (Thompson, Johnson, Lavis, and Williams, 1974), but the significance of this report is unclear since secretin has not yet been shown to increase cyclic GMP levels in any tissue. Thus, although not convincingly established, some hormones may increase particulate guanylate cyclase activity in a manner similar to that by which hormones increase adenylate cyclase activity. Alternatively, some hormones may increase soluble guanylate cyclase activity with mediating factors other than Ca++ being involved, or hormone-receptor interaction at the plasma membrane could conceivably induce a dislocation and change in effective activity of a reversibly bound, membrane-associated guanylate cyclase. Elucidating which or how many of these possibilities are operative will require thorough study and understanding of the fundamental behavior and properties of soluble and particulate guanylate cyclase activities.


Relationship between cyclic AMP metabolism and inotropic response of perfused rat hearts to phenylephrine and other adrenergic amines. A positive correlation between inotropic response and increment of cyclic AMP levels and labeling (from 14-C-adenine in the perfusate) was found after isoprenaline, dopamine, and phenylephrine alone, and after isoprenaline in the presence of chlorpromazine. 2. a lack of correlation between contractile activity and cyclic AMP levels and labeling was found when the hearts were exposed to phenylephrine in the presence of propranolol. The contractile activity increased, but the parameters for cyclic AMP did not change. These resuls together with findings by other workers indicate that  the inotropic response to beta-stimulation always is associated with cyclic AMP accumulation, while the inotropic response to alpha-stimulation (when present) is not correlated to cyclic AMP elevation. Other differences between alpha- and beta-adrenergic effects on heart (course of development of the inotropic response, effect of theophylline on the response, effects on relaxing processes, action potential, and on refractory period) also indicate different mechanisms of action. On the basis of the available data we suggest as a conclusion that the inotropic response after alpha-adrenergic stimulation does not involve cyclic AMP as a mediator whereas beta-adrenergic effects are mediated by cyclic AMP. This means that the naturally occurring adrenergic amines norepinephrine and epinephrine, both of which are able to stimulate both alpha and beta myocardial receptors (twenzel and Su, 1966), elicit the inotropic response through two different mechanisms. Usually the beta-adrenergic effect on the heart is the more important. The contribution from alpha-stimulation, however, may increase under certain conditions: hypothyroidism or proplythiouracil treatment per se (Nakashima et al., 1971) or hypothermia (Kunos and Szentiványi, 1968; Buckley and Jordan. 1970; Benfey et al., 1973; Kunos, Yong, and Nickerson, 1973; Nickerson, 1973). Thus a single physiologic response may be mediated by more than one mechanism. Multiple mechanisms of action for one agent might have more general biologic significance; e.g., they may serve to maintain the responsiveness of a tissue under various conditions.


Control of calcium transport in the myocardium by the cyclic AMP-Protein kinase system. At least three mechanical changes characterize the response of cardiac muscle to agents that enhance cyclic AMP production. In common with other inotropic interventions, tension is augmented and the rate of tension rise is increased. The third response, acceleration of the rate of relaxation, is characteristic of the actions of beta-adrenergic agonists. These mechanical effects can be attributed to changes in (1) the amount of Ca2+ released during systole, (2) the rate of Ca2+ release at the onset of systole, and (3) the rate at which Ca2+ is reaccumulated by the sarcoplasmic reticulum at the end of systole. The ability of cyclic AMP-dependent protein kinases to phosphorylate the cardiac sarcoplasmic reticulum in vitro parallels stimulation of both Ca2+ transport and Ca2+-activated ATPase. The phosphoprotein formed in the presence of cyclic AMP and protein kinase has the chemical characteristics of a phosphoester, contains mostly phosphoserine, and has an electrophoretic mobility in SDS polyacrylamide gels that corresponds to a protein of 22,000 daltons. This 22,000-dalton protein, tentatively named phospholamban, thus differs from the acyl phosphooprotein formed by the Ca2+-transport ATPase, which as an apparent molecular weight of 90,000 to 100,000 daltons. Phospholamban has not been found in fast skeletal muscle, nor is Ca2+ transport accelerated by cyclic AMP and protein kinase in sarcoplasmic reticulum from these muslces which do not respond to beta-adrenergic agonists with accelerated relaxation. It thus appears likely that phosphorylation of phospholamban correlates both with an increased rate of Ca2+ transport by cardiac sarcoplasmic reticulum in vitro and accelerated relaxation in the intact myocardium. Preliminary findings are consistent with the view that phosphorylation of phospholamban may be related to other actions on Ca2+ fluxes brought about by agents which activate adenylate cyclase in the myocardium, but these interpretations must remain speculative pending more definitive studies.


Role of cyclic AMP in the actions of catecholamines on hepatic carbohydrate metabolism. Studies using perfused rat liver and isolated liver parenchymal cells show that low concentrations of epinephrine, norepinephrine, and phenylephrine can activate glycogenolysis and gluconeogenesis by a mechanism mediated by alpha-adrenergic receptors and not involving accumulation of cAMP. The glycogenolytic and gluconeogenic activites of epinephrine, norepinephrine, and phenylephrine are inhibited by the alpha-adrenergic receptor antagonists phentolamine and dihydroergotamine, but are negligibly affected by the beta-adrenergic receptor antagonist propranolol. Epinephrine, norephinephrine, and the beta-adrenergic receptor agonists isoproterenol, soterenol, and salbutamol increase cAMP accumulation; and this effect is antagonized by propranolol. Isoproterenol, soterenol, and salbutamol activate glycogenolysis and gluconeogenesis, but are less effective than epinephrine or norepinephrine. The data  are interpreted as indicating the existence of both alpha- and beta-adrenergic receptors in rat liver. Activation of the alpha-adrenergic mechanism appears to be more important than the beta-adrenergic receptor-cAMP system in the physiologic effects of epinephrine and norepinephrine on carbohydrate metabolism in rat liver.


Glucagon and adenylate cyclase: binding studies and requirements for activation. Solubilization of myocardial adenylate cyclase abolished responsiveness to glucagon and catecholamines, two of the hormones which activate the membrane-bound enzyme. Adenylate cyclase freed of detergent by DEAE-cellulose chromatography continues to remain unresponsive to hormone stimulation. However, adding purified bovine brain phospholipids--phosphotidylserine and monophosphatidylinositol--restored responsiveness to glucagon and catecholamines, respectively. 125-i-glucagon binding appeared to be independent of phospholipid, since equal binding was observed in the presence or absence of detergent and in the presence or absence of phospholipids. Chromatography of the solubilized preparation on Sephadex G-100 WAS CHARACTERIZED BY 125-I-glucagon binding and fluoride-stimulatable adenylate cyclase activity appearing in the fractions consistent with the void volume, suggesting a molecular weight greater than 100,000 for the receptor-adenylate cyclase complex. Prior incubation of the binding peak with 125-I-glucagon and rechromatography of the bound glucagon on Sephadex G-100 shifted its elution to a later fraction consistent with a smaller-molecular-weight peak. The molecular weight of this material was 24,000 to 28,000, as determined by SDS polyacrylamide gel electrophoresis. The latter findings are consistent with a dissociable receptor site for glucagon on myocardial adenylate cyclase.


Regulation of cyclic AMP content in normal and malignant brain cells. Pharmacologic characterization of the neurotransmitter-sensitive cyclic AMP-second messenger systems of brain has proven to be a complex and difficult endeavor. At least two types of receptor appear to be involved in the mediation of the effects of NE on cyclic AMP content. One of these receptor systems appears to mediate the potentiation by NE of the effect of adenosine of cyclic AMP accumulation. The cellular heterogeneity of brain has retarded the determination of the mechanism underlying the synergistic interaction of catecholamines and adenosine. An attempt to use clonal cell lines to examine the action of NE and adenosine on cyclic AMP content has resulted in the demonstration that adenosine acts in a hormone-like fashion to stimulate adenylate cyclase activity. However, the studies did not shed light on the mechanism of synergism. An increasing  number of reports are appearing which support the idea that the responsiveness of cells to neuronally released NE may involve adaptive changes in the responsiveness of the cyclic AMP-second messenger system which compensate for chronic over- or underproduction of the first messenger, NE. Evidence was presented that such a regulatory process may be operative in rat cerebral cortex. Our studies of catecholamine-induced loss of responsiveness in human astrocytoma cells have led us to the conclusion that the loss in the capacity of the cells to accumulate cyclic AMP is a result of a loss in the capacity to synthesize cyclic AMP. However, it is probable that different cells make use of different mechanisms (e.g., changes in phosphodiesterase activity) to regulate their ability to respond to hormones or neurotransmitters. The physiologic importance of this level of regulation of responsiveness to hormones is not known at this time.


Development of the implantation chamber in the pregnant bitch. Uteri taken from 25 bitches at various times during the early stages of pregnancy were studies cytologically to determine how the implantation chamber developed and how fetal-maternal relations were established. On day 13 after the end of estrus, knobs of trophoblastic syncytium formed and became wedged between cells of the uterine luminal epithelium. The syncytium quickly spread along the uterine lumen and into the mouths of the glands, dislodging and surrounding maternal cells. As invasion continued trophoblastic villi, consisting of cores of cytotrophoblast covered by a continuous layer of syncytium, penetrated deeper into the endometrium. The syncytium spread to surround maternal vessels and decidual cells. By day 26 the trophoblast had extended down to the large lacunae. Here syncytial trophoblast covering tips of the villi degenerated, leaving cytotrophoblast exposed to the necrotic zone. These cells possessed characteristics of absorbing cells. Hematomas were formed by focal necrosis of fetal and endometrial tissue at the poles of the implantation sites. Large pools of extravasated blood accumulated and red blood cells were phagocytized by surrounding trophoblastic cells. Therefore, the endotheliochorial relationship in the canine placenta appeared to be established by syncytial trophoblast invading a cellular endometrium. In the necrotic zone and hematomas, cellular trophoblast may have lost its syncytial covering, but elsewhere maternal vessels and decidual cells in the placenta were in direct contact only with syncytial trophoblast.


Light and electron microscopic studies of postcapillary venules in developing human fetal lymph nodes. Developing lymph nodes from 30 human fetuses with crownrump lengths (CRL) of 38 mm (8.7 wk) to 245 mm (26 wk) were studied by light and electron microscopy. Blood vessels that appear to be postcapillary venules (PCV) are present in nodes of 47 mm CRL and older fetuses. These venules first appear in nodes whehn the nodal population of lymphocytes is sparse. In these early nodes PCV are distributed randomly and consist of a low endothelium, underlying basal lamina and incomplete pericyte sheath. Early nodal PCV are distinguised from other nodal blood vessels by the presence of lymphocyte diapedesis and several luminal lymphocytes. In the late stages of nodal development PCV are the more common non-capillary blood vessel and appear in the parenchyma near the periphery of the node. Late nodal PCV are generally characterized by a cuboidal endothelium that is rich in Golgi apparatus, lysosomes and Weibel-Palade bodies. The lumen and wall of late nodal PCV contain lymphocytes. The relationship between the development of the parenchyma of fetal nodes and the appearance and activity of PCV, the passage of lymphocytes through the PCV wall and the fine structure of developing PCV are described. It is suggested that the lymphocytes that first appear in developing nodes, and the majority of the lymphocytes found in late nodes, migrate to the node via the blood vascular system and enter the nodal parenchyma by passing across PCV endothelium.


Quantification of plasma cholesterol and triglyceride levels in hypercholesterolemic subjects receiving ascorbic acid supplements. In order to assess the possible effects of ascorbic acid on plasma cholesterol and triglyceride levels and plasma lipoprotein composition, nine hypercholesterolemic subjects were treated with oral ascorbic acid (4 g/day) for 2 months. The data demonstrate: 1) no significant change in plasma cholesterol or triglyceride levels; 2) no significant change in the cholesterol or triglyceride concentrations of the major lipoprotein classes; and 3) the unexpected appearance of extra pre-beta bands on lipoprotein electrophoresis by the end of the ascorbic acid treatment period.


Diet and intestinal enzyme adaptation: implications for gastrointestinal disorders. Recent studies have demonstrated that the human intestinal enzymes of carbohydrate digestion and metabolism can be regulated by dietary sugars. These studies have utilized direct assay of intestinal mucosal enzyme activity. Mucosa has been obtained by the use of peroral jejunal biopsy techniques which provide 10-15 mg of mucosa in a safe, simple and reproducible manner. Dietary sucrose, as compared to dietary glucose, increases the activities of the jejunal disaccharidases, sucrase and maltase, but not lactase. Fructose reproduces the sucrose effect and appears to be the active principle in the sucrose molecule. Lactose deprivation or lactose feeding does not alter lactase activity. Fructose has been useful in treating one patient with sucrase-isomaltase deficiency. Jejunal glycolytic enzyme activities are also regulated by dietary sugars. Certain enzymes are highest with specific dietary carbohydrates, lower with other sugars and lowest on a carbohydrate-free diet. The regulation of human jejunal glycolytic enzyme activity takes place in hours, whereas the change in disaccharidase activity occurs in 2-5 days. The mechanism of this regulation is not known. Additional investigations have shown that jejunal glycolytic enzyme activities but not the disaccharidases are controlled by oral folic acid as well. This effect occurs within 1 day also. The mechanism is unknown. Large doses of folate have been of benefit in a few patients with certain glycolytic enzyme deficiency states. Preliminary studies have demonstrated that selected patients with chronic undiagnosed intestinal disorders fail to manifest an adaptive response of their jejunal glycolytic enzyme activities to dietary sugars. This condition has been termed a "maladaptation syndrome.".


Cell membrane receptors for polypeptide hormones. Application to the study of disease states in mice, and men. Recent studies have suggested that the first step in the action of polypeptide hormones and catecholamines is binding to surface membrane receptor sites. This interaction can be studied directly using radioactively labeled hormones and a variety of receptor preparations. With these radioreceptor assay technics, it has been possible to define structure-activity relationships and to assay plasma hormones. These studies have also been used to define alterations in the hormone receptors in disease that lead to peripheral resistance to hormonal action and to characterize the factors that regulate receptor concentrations.


Comparisons of major cell-surface proteins of normal and transformed cells. Transformation of the chick fibroblast surface has been studied in cells infected with Schmidt-Ruppin Rous sarcoma virus and the temperature-sensitive mutant of this virus, TS-68. Major findings following transformation induced by a shift from nonpermissive (41 C.) to permissive (36 C.) temperature in TS-68 infected cells were: (1) rapid cessation or slowing of the synthesis of a protein, M.W. 100-200,000, localization uncertain; (2) cessation or slowing of the synthesis of a plasma membrane protein, M.W. 45,000, within 2-4 hours; (3) cessation or slowing of the synthesis of a large trypsin- and collagenase- sensitive protein (M.W. greater than 200,000) only after an extended period of morphologic transformation. In addition, increased quantities of type-specific viral antigen in the membranes of infected cells were observed in TS-68-infected cells at 41 compared with 36 C.


Composition of membranes of cells transformed by tumorigenic DNA and RNA viruses. Continuous cell lines obtained from mouse and hamster embryonic tissue and human lung contain a homologous series of acidic glycolipids known as gangliosides. These complex lipids are highly concentrated in plasma cell membranes. The ganglioside pattern becomes greatly simplified when cells are transformed by tumorigenic DNA and RNA viruses. Non-transforming viruses and lytic infection of cells by tumorigenic viruses without neoplastic tranformation do not cause changes in gangliosides. The alterations of ganglioside composition are the result of specific blocks in the biosynthesis of these lipids. The significance of these impairments of membrane glycolipid formation and the nature of the regulatory site(s) involved are considered with regard to the altered social behavior of transformed cells.


Multilamellar cytosomes in a particular form of late-infantile amaurotic idiocy. There is a particular form of late-infantile amaurotic idiocy in which no clear chemical-pathological or unique enzyme abnormalities have been identified to date. A distinctive morphological feature has been recognized on electron microscopical examination of tissues from these patients, which has been descriptively labeled with various terms, including "multilamellar cytosomes" (MLC). Illustrations of MLC in a patient with this late-infantile form of cerebroretinal degeneration show their reactivity with the periodic acid-silver methenamine reaction for glycoproteins. The MLC are shown to be morphologically identical in cerebral tissue obtained at biopsy, in the same tissue obtained three years later at autopsy, and in skeletal muscle.


Effects of gastrointestinal hormones on vascular smooth muscle. The effects of the gastrointestinal hormones secretin and cholecystokinin (CCK) were investigated on rat portal-vein mechanical activity. Doses of 0.03-0.60 units/ml Boots secretin; 0.18-0.60 units/ml pure, natural secretin; and 0.12-0.60 units/ml CCK elicited dose-dependent decreases in amplitude and simultaneous increases in frequency of spontaneous contractions that were unaffected by alpha-or beta-adrenergic or cholinergic blocdade. These effects were mimicked by isoproterenol, theophylline, and cyclic AMP, and all agents tested were also effective in reducing K-or norepinephrine-induced tension. Additionally, subthreshold doses of CCK and secretin elicited responses when given in combination, and the effects of the hormomes at all doses were potentiated by theophyline, indicating an interaction berween these hormones on rat portal vein thatmay involve cyclic AMP. It is suggested that part of the mechanism of hormone-induced mesenteric vasodilatation may involve a direct relaxing effect of the hormones on vascularsmooth muscle.


Cholagiocarcinoma in a patient previously given thorotrast. The long-term effects of thorotrast administration include local granulomas, blood cell abnormalities, and cancer. To date, more than 100 cases of liver cancer have been reported in patients previously given this radioactive contrast material, with a latency period of approximately 22 years. We report such a case; the patient not only had a thorotrast-induced cholangiocarcinoma, but also showed striking morphologic changes in the red blood cells secondary to splenic dysfunction.


Crohn's disease of the colon. III. Toxic dilatation of the colon in Crohn's colitis. In a group of 160 patients with Crohn's disease involving the colon, there were seven patients with toxic dilatation, four with granulomatous colitis and three with ileocolitis, all successfully treated without mortality. This complications is more common than previously recognized in Crohn's colitis. In Crohn's disease, toxic dilatation is less likely to proceed to perforation of the bowel, because of the nature of the pathology and is more likely to respond to conservative measures: intubation, with decompression, corticotropin, steroids and  high-dose antibiotic administration. Although patients do recover from this life-threatening complication with conservative management, the majority of patients, if not all, will ultimately come to surgical excision of the colon. If surgery is mandatory, it should be carried out early, rather than late, in the patient who is failing to respond to medical therapy, certainly before the development of perforation, massive hemorrhage, or gram negative sepsis with shock. The surgical therapy will depend upon the state of the bowel at laparotomy. Thus, an intact bowel in a young patient, would favor subtotal colectomy or proctocolectomy; a sealed perforation, a diverting ileostomy with skin level colostomy decompression as suggested by Turnbull and a free perforation, the minimum adequate procedure which will tide the patient over the early postoperative period. Diverting ileostomy alone has been effective in two of our patients but should be avoided in ulcerative colitis. The critically ill patient with the ominous finding of "disintegrating colitis" and multiple leaks, will require nothing less than total radical excision of the diseased bowel in the hope of immediate salvage.


The sudden infant death syndrome and epidemic viral disease. A study was done to investigate the relationship between the sudden infant death syndrome (SIDS) and epidemic respiratory viral disease among hospitalized children under 18 months of age. During the 42 month-period of this study, there were 778 sudden infant deaths in Chicago and 3244 hospital admissions of children under 18 months for respiratory disease. Four outbreaks of respiratory syncytial (RS) virus infections, three outbreaks of influenza A virus infections, and several small clusters of parainfluenza virus infections occurred during the course of this study. Influenza A was the only virus infection found to have a statistically significant association with SIDS. Although environmental temperature was also significantly correlated with SIDS, the association with influenza A virus infection was independent of this temperature effect and neither association was strong.


Serologic characterization and sero-epidemiologic studies on acute hemorrhagic conjunctivitis (AHC) virus. Serologic and sero-epidemiologic characteristics of AHC virus infection were studied by neutralization test (NT). Four-fold or greater virus neutralizing (VN) antibody response was demonstrated to the Japanese isolate of AHC virus (the J 670/71 strain) in 77.3% and 66.7% of paired sera from clinical AHC patients in Japan (1971-1973) and Tunisia (1973). The four patients from Indonesia studied in 1972 showed similar antibody response. Cross-neutralization tests of AHC virus isolated in Japan (1971), Taiwan (1971), Hong Kong (1971), Thailand (1972), Indonesia (1972), Singapore (1972), Morocco (1971) and England (1971) with three kinds of antisera prepared against Japanese, Hong Kong and Moroccan AHC virus isolates indicated their antigenic identity. However, isolates from Sinapore in 1970 (Singapore 70 virus) were not neutralized with the AHC virus antisera mentioned above: Singapore 70 virus constitutes another antigenic type, to which, however, no VN antibody rise was found in paired patients' sera from Japan, Tunisia and Indonesia. Thus, no serologic evidence supporting an etiologic role of this virus group in the development of AHC was found. Although cross-tests using monospecific antisera suggested some cross-relation between AHC and both echovirus type 4 (E4) and coxsackie A (CA), type 19, no serologic relationship between AHC and these viruses was found. Sera from healthy individuals collected before and after AHC outbreaks were tested for VN antibody against AHC virus in Japan and two epidemic foci, Ghana and Indonesia. Before the epidemic, 80 to 90% of the people lacked antibody in the three countries, but 39.7% and 45.2% of inhabitants posessed VN antibody of 1:8 or over in Ghana and Indonesia after the outbreak. In Japan, however, only a slight increase was found in VN antibody prevalence afterwards. Serologic study showed that 41.5% of horse sera were VN positive at dilutions of 1:8 or more; many cattle sera also had a low VN titer but few cynomologus monkey sera had VN activity.


Serologic profile of children in a Mexican highland community: prevalence of complement-fixing antibodies to Mycoplasma pneumoniae, respiratory syncytial virus and parainfluenza viruses. The study investigated the prevalence of antibodies to five leading agents of childhood respiratory disease in the county of Huixquilucan, Mexico. Tests of sera from a random sample of children between 3 months and 18 years of age confirmed serologically the presence of Mycoplasma pneumoniae, respiratory syncytial (RS) virus and parainfluenza 1, 2 and 3 viruses in this relatively isolated highland community. Highest overall antibody frequency of 64.2% was seen for parainfluenza 3, and antibody to this virus was acquired early in life. Antibody to M. pneumoniae was least prevalent among children surveyed; this rate was 15.5% overall. This was only slightly below the prevalence rates for antibodies to RS virus and parainfluenza 1 and 2 viruses, which had intermediate frequency rates of 23%, 32%, and 22.9%, respectively. The relatively low prevalence of antibody to RS virus was unexpected. Differences in prevalence rates in regard to location of residence or family size were insignificant. Statistically significant differences in age-specific antibody prevalence rates in respect to sex were noted only for the 5- to 9-year-old group to M. pneumoniae and to parainfluenza 3.


Progress in hepatitis research.

Progress in hepatitis research. A brief account of recent developments and current nomenclature regarding the viral agents of hepatitis A and B is presented.


Physiologic significance of 17beta-estradiol binding in the rabbit Fallopian tube. Rabbit Fallopian tube contractility was recorded in vitro during perfusion with either Locke's solution or that solution containing CN-55, 945-27 (CN; or CI-628), a nonsteroidal estrogen antagonist (Endocrinology 79: 153, 1966). Contractility was inhibited and 17beta-estradiol (E2) displaced from both its cytoplasmic (8S) and nuclear (4S) receptors in the presence of the above agent. These effects result from a direct interaction between CN and the E2-receptor complexes. Two types of evidence show the specificity of the foregoing responses: (1) nonspecific binding of E2 to serum proteins was unaffected by the antagonist and (2) CN had no effect on contractility of a nontarget tissue, i.e., rabbit ileum. In addition, Fallopian tube contractions induced by strong electrical stimulation of K-depolarized tissues (i.e., in the absence of normal ionic gradients) were inhibited by CN and a decrease in the binding capacities of 8S and 4S receptors was again observed. Thus, antagonism of specific E2 binding inhibits the contractile mechanism at a level other than the cell membrane. These observations, and additional findings concerning the reversibility of CN action, indicate that E2 binding is essential for contractility of the rabbit Fallopian tube.


Intracellular Treponema pallidum in cells of a syphilitic lesion of the uterine cervix. In an electron microscopic study of biopsy tissue from an ulcer of the human cervix uteri, organisms morphologically identical to T. pallidum were observed. Large numbers of the organisms were seen lying among collagen fibrils and between cells. T. pallidum was seen within vacuoles of some fibroblasts and epithelial cells. Within the nucleoplasm of several cells, treponemes were observed, surrounded by a clear zone which might represent a mucopolysaccharide outer layer. The siameter of the organisms was 171 nm., and the diameter of the external axial filaments was 25 nm., dimensions similar to those reported by others for T. pallidum.


Infectious mononucleosis preceding acute myelomonocytic leukemia. The patient discussed in this report had acute myelomonocytic leukemia (AMML) associated with infectious mononucleosis. Infectious mononucleosis (IM) in association with acute leukemia (AL) has provided clinical investigators an active area of speculation for several years. Nineteen cases are presented for review in Table 1. The present case represents the second case of AMML to be associated with IM. It is the first case of a granulocytic leukemia reported in which active IM is occurring simultaneously with the onset of AL. Also, it represents the first case of associated IM-AL in which the diagnosis of IM is supported by lymphocyte culture data. This may represent a mechanism for additional investigation in future cases.


Skeletal muscle respiratory capacity, endurance, and glycogen utilization. This study was undertaken to evaluate the relationship between physical performance capacity and the mitochondrial content of skeletal muscle. Four groups of rats were trained by means of treadmill running 5 days/wk for 13 wk. One group ran 10 min/day, a second group ran 30 min/day, a third group ran 60 min/day, and a fourth group ran 120 min/day. The magnitude of the exercise-induced adaptive increase in gastrocnemius muscle respiratory capacity varied over a twofold range in the four groups. There were significant correlations between the levels of three mitochondrial markers (cytochrome c, citrate synthase, respiratory capacity) in the animals' gastrocnemius muscles and the duration of a run to exhaustion. There was also a significant correlation between the amounts of glycogen remaining in liver and skeletal muscle after a 30-min-long exercise test and the respiratory capacity of the animal's leg muscles. These findings are compatible with the interpretation that a close relationshiop exists between skeletal muscle mitochondrial content and the capacity to perform endurance exercise.


Hypothalamic cells in an area mediating ACTH release respond to right atrial stretch. The right atrium has been shown to mediate release of ACTH in response to small hemorrhage. Therefore, we sought to identify hypothalamic neurons (cat, alpha-choloralose 35-45 mg/kg) wholse firing rates were modulated by stretch of the right atrial-vena caval junction. Stretch was produced by intermittent inflation of a balloon implanted between the pericardium and the right atrium. In some cases, carotid arteries were occluded bilaterally. Previous results indicate that electrical stimulation of the medial-dorsal hypothalamus (MDH) causes prompt changes in ACTH, but stimulation in the lateral hypothalamus does not cause changes. Of 64 cells recorded in the MDH, 41 responded to atrial stretch. However, only 2 of 18 cells outside this region responded. Six cells were studied with and without carotid occlusion. Their response was dimished or absent in the absence of carotid occlusion. The results suggest that integration of afferent signals from cardiovascular receptors occurs in the MDH and that the MDH may participate in neural mediation of ACTH in response to hemodynamic changes.


Regulation of liver diphosphofructose phosphatase of the rat and hamster. The regulatory properties of D-fructose-1, 6-diphosphate 1-phosphohydrolase (FDPase, EC 3.1.3.11) derived from livers of the albino rat and the golden Syrian hamster have been studies. The comparative analysis of the kinetics of saturation by fructose-1, l-diphosphate (FDP) and inhibition by FDP and by the allosteric ligand adenosine 5'-monophosphate (AMP) revealed differences that may have physiological implications. The inhibition studies pointed out that liver phosphatase is under a significantly less stringent regulation in the hibernating rodent. The concentration of AMP required to produce a 50 percent inhibition was 549.5 muM at 37 degrees C, 21.4 muM at 10 degrees C, and 9.2 muM at 5 percent C for the rat; 1,122 muM at 37 degrees C, 34.3 muM at 10 degrees C, and 20.4 muM at 5 degrees C for the hamster. Urea (0.67 M) simulates the effect of low temperature, potentiating the AMP inhibition. At 5 degrees C and 4 M urea, 75 percent loss of activity was found for the rat FDPase in contrast to a 25 percent loss for the hamster. At 37 degrees C, 4 M urea had practically no effect on the FDPase of both species.


Afferent aortic nerve fibers with their pathways in cardiac sympathetic nerves. The effect of mechanical and chemical stimulation on activity of afferent aortic nerve fibers with pathways in the cardiac sympathetic nerves has been examined. Action potentials were derived from the second or third thoracic communicating ramus of the left side of anesthetized dogs. Thirty myelinated and 19 unmyelinated fibers responded to tapping the ascending aorta, aortic arch, and descending aorta. Both groups of fibers also responded to a rise as well as to a fall in aortic pressure. Spontaneous discharge of myelinated fibers was related to aortic pressure pulse whereas that of unmyelinated fibers was related to respiration. Asphyxia caused excitation of unmyelinated fibers but not of myelinated fibers. Both groups of fibers responded to topical application of lactic acid. Mechanical and chemical stimulation of the aorta after vagotomy caused a rise in systemic blood pressure and extension of the limbs. The results indicate the existence of afferent aortic fibers in the cardiac sympathetic nerves that cause circulatory and somatic responses.


Reduced nicotinamide adenine dinucleotide and depolarization in neurons. Activity of frog dorsal root ganglion neurons, evoked by dorsal root stimulation under aerobic conditions, produced a change in the level of reduced nicotinamide adenine dinucleotide (NADH) on the surface of this tissue. There was a decrease of NADH (oxidation), followed by an increase of NADH (reduction). Both changes were dependent on previous activity, and a critical time is required in order to observe a similar response. At the frequency and duration of stimulation used here, each stimulus evoked neuron depolarization as shown by recording from single cells. The NADH oxidation occurred in the respiratory chain and it was selectively blocked by Amytal. The NAD reduction was attributed to 3-phosphoglyceraldehyde dehydrogenase activity and it was blocked by iodoacetate. The NADH oxidation and NAD reduction were attributed to respiratory chain activity and aerobic glycolysis, both activated and deactivated by changes of phosphate potential (ATP/ADP + Pi). A low phosphate potential activates the respiratory chain and glycolysis; a high phosphate potential deactivates the respiratory chain and glycolysis.


Effects of thyroid hormone administration on skeletal muscle mitochondria. The effects of thyroid hormone administration on the levels of a number of mitochondrial markers were measured in skeletal muscle and liver of normal rats. Injection of 18 mug of L-thyroxine (T4) per 100 g body wt every 4th day for 3 wk had no effect on the concentrations of cytochrome c, on citrate synthase activity, or on respiratory capacity of skeletal muscle. Injection of 200 mug of L-triiodothyronine (T3) daily for 5 days, or feeding 23 mg T4 and 7 mg T3/kg of diet for 2 wk, resulted in thyrotoxicosis and large increases in the activity of hepatic alpha-glycerophosphate dehydrogenase and other mitochondrial markers; however, the levels of activity of mitochondrial marker enzymes in gastrocnemius and quadriceps muscles were not significantly changed. Only when rats were fed 3 mg T4 and 1 mg T3/kg diet for a 6-wk period did we observe an increase in skeletal muscle mitochondrial markers. Thus, thyroxine treatment must be sufficiently prolonged if it is to be used as a tool for studying skeletal muscle mitochondrial biogenesis.


Arterial water, cations, and norepinephrine in early and late renovascular hypertension. In dogs made hypertensive renal artery stenosis and contralateral nephrectomy, the arterial and myocardial tissue content of water, cations (sodium, potassium, magnesium, calcium), and norepinephrine was measured 20 and 60 days after the operation. Hypertensive animals autopsied at the earlier stage of hypertension had significantly lower (-25 to-50%, P smaller than 0.01) arterial norepinephrine than either the sham or nonhypertensive animals. The water and cation content of arteries was unchanged, but aortic tissue contained significantly more water, sodium, potassium, and magnesium than the nonhypertensives. Hypertensives autopsied at 60 days also had lower arterial norepinephrine content than sham and nonhypertensive animals, but this refuction was not significant. Their arterial and aortic tissue contained more water and cations than the nonhypertensive, sham, unoperated, and unilaterally nephrectomized dogs. It is suggested that in the initial stage of renovascular hypertension the arterial norepinephrine content is reduced significantly, whereas at a later stage the arterial water and cations may be involved in the maintenance of an elevated blood pressure.


Transport of Na, Cl, and water by the rabbit corneal epithelium at resting potential. Theophylline (1 mM) produced a net transport of Na and Cl from aqueous humor to tears (.02-.04 mumol/cm2 h) in the isolated rabbit cornea denuded of endothlium and in the presence of normal resting potential (25-35 mV). The active transport of Na (tears to aqueous) and of Cl (aqueous to tears), estimated with the Goldman constant-field equations, was confirmed. A 10 degrees C rise in temperature produced changes close to those predicted for passive processes in both unidirectional fluxes of Na and in the tears-to-aqueous flux of Cl, but not for the aqueous-to-tears flux of Cl. Theophylline treatment doubled Cl permeability but did not significantly affect Na or urea permeability, suggestingspecificity of affect. In separate experiments it was shown that stromal thinning occurred in previously swollen corneas when the endothelium was blocked by silicone oil and the epithelium was treated with theophylline. These findings provide further support for the argument that the mammalian epithelium could have an active role in the regulation of corneal thickness in situ.


Mechanism of cholecystokinin inhibition of lower esophageal sphincter pressure. The purpose of this study were 1) to quantify the lower esophageal sphincter (LES) response to intravenous cholecystokinin (CCK) in both man and the opossum in vivo, 2) to characterize the interaction of CCK and gastrin on circular muscle of the LES, and 3) to determine the site of action of CCK on LES muscle. In both man and the opossum LES pressure was decreased significantly by either constant intravenous infusion or bolus injection of CCK. In vitro dose-response curves to gastrin I, CCK, and the octapeptide of CCK (OP) demonstrated that both CCK and OP were partial agonists on the LES muscle. Both CCK and OP contract LES muscle at lower threshold doses, but give smaller maximum responses than gastrin I. The maximum response of LES muscle to CCK was antagonized only by atropine and tetrodotoxin, but not by other antagonists, suggesting that CCK contracts LES muscle by acetylcholine release. In vitro studies on LES muscle showed that CCK selectively antagonized the effect of gastrin I, but not other agonists. These studied suggest that CCK reduces LES pressure in vivo by inhibition of the endogenous gastrin effect.


Effect of amino acid meals on hepatic alpha-aminoisobutyrate transport and cyclic AMP. Within 1.5 h after force-feeding rats one meal of enzymatic hydrolysates of casein, gelatin, lactalbumin, or yeast, alpha-aminoisobutyric acid (AIB) transport in liver slices was stimulated two- to threefold. A complete amino acid mixture also increased AIB transport. Of the 15 amino acids or derivatives tested individually, the dispensable amino acids, especially glycine and alanine, were more stimulatory than the essential amino acids; feeding a mixture of amino acids lacking glycine and alanine increased AIB uptake only slightly. The effects were significantly greater in meal-fed than in ad libitum-fed rats. Increased hepatic concentrations of cyclic AM were usually associated with the increase in AIB transport. Feeding glucose inhibited the increases in transport and cyclic AMP concentration induced by casein hydrolysate or in the stimulation of AIB transport by dietary amino acids. The increases in AIB uptake appeared unrelated to the exchange of endogenous amino acids with medium AIB.


Granular cell myoblastoma of the cystic duct. A case associated with hydrops of the gallbladder. The first known case of true hydrops of the gallbladder that resulted from granular cell myoblastoma is reported. Granular cell tumors are benign multicentric tumors that should be treated by total excision so that free margins are assured. These tumors are rarely found in the biliary tract; however, they may cause complications of biliary tract obstruction.


Alopecia, ascites, and incomplete regeneration after 85 to 90 per cent liver resection. A nineteen year old female underwent 85 to 90 per cent partial hepatectomy to treat a minimal deviation hepatoma. Observations afterwards suggested that the limit of resection compatible with survival had been reached. She recovered perfect health after many months, although liver regeneration was not complete. Severe but eventually reversible alopecia and ascites developed postoperatively, undoubtedly as a complication of the massive hepatic resection.


Intraoperative physiologic monitoring and management during hepatic lobectomy using the liver isolation-perfusion technic. The high operative mortality of major hepatic resection for tumor can be improved by a technic of resection using complete vascular isolation and hypothermic perfusion of the liver. Complete clamping of the portal vein, vena cava, and hepatic artery was necessary and well tolerated. Major physiologic, biochemical, and coagulation changes, however, can occur with this technic that requires close monitoring by the anesthesiologist. With astute observation and prompt corrective measures when indicated, these changes can be minimized to enable a safe and smooth resection to be carried out.


Thyrolymphography.

Thyrolymphography. The present investigation shows that the capacity of thyrolymphography for demonstrating the morphologic features of various thyroid lesions is not inferior to that of isotope scanning. On the contrary, lymphography is an effective procedure for detecting small nodules that remain undetected by other methods. Contrast medium is more rapidly distributed to lymph nodes in patients with diffuse goiter and also in patients pretreated with Lugol's solution. Finally, it is shown that the contrast medium employed (Lipiodol Ultra Fluid) remains in the lymph nodes for a period of several months.


Total pancreatectomy with primary mesenteric vascular reconstruction. Total pancreaticoduodenectomy with resection of the superior mesenteric artery and vein was performed in a sixty-two year old man with islet cell carcinoma of the pancreas. Arterial reconstruction was accomplished with an aortomesenteric interposition Dacron graft. The venous system was reanastomosed primarily, the infrapancreatic superior mesenteric vein to the portal vein in the hepatic hilum. This procedure, although it did not result in long-term survival, demonstrates the technical feasibility of wide regional resection for tumors of the pancreas impinging upon the mesenteric vasculature in the region of the neck and uncinate process.


Parotid gland tumors. Clinicopathologic study. The clinical findings and histologic features of 154 parotid gland tumors treated at the Health Sciences Centre, Winnipeg, Canada, from 1957 to 1970 were reviewed. The study illustrates a wide variation in the natural history and degree of malignancy of parotid tumors. Approximately 80 per cent were benign; among these, mixed tumors were by far the commonest. For most benign tumors, the recommended surgical treatment is wide local excision with an adequate margin of normal glandular tissue, except when subtotal or conservative total parotidectomy is required because of the size or location of the tumor. The facial nerve should be visualized and preserved. This approach prevents recurrences and minimizes facial nerve injury, since risk of neural injury increases with the extent of the surgical procedure. Total removal of the gland on the basis of the multicentricity or malignant transformation of these tumors is not supported by the findings of this study. For malignant tumors a more radical procedure, that is, subtotal or total parotidectomy with or without node dissection, with sacrifice of the facial nerve if necessary is advisable, depending on the type and anatomic location of the tumor.


Electron microscopic studies on zonular fibers. II. Changes of the zonular fibers after the treatment with collagenase, alpha-chymotrypsin and hyaluronidase. Adult rabbit zonular fibers maintained in their native condition were treated with collagenase, alpha-chymotrypsin and hyaluronidase, and were observed with the electron microscope. The results obtained were as follows: 1. Collagenase digested the lens capsule, but not the zonular fibers. 2. Long time collagenase action obscured the cell membrane of the lens epithelium and the basal lamina of the ciliary epithelium. 3. Washing with 0.9% NaCl increased the collagenase action on the lens capsule. 4. Alpha-chymotrypsin digested the zonular fibers and the zonular lemalla, but not the lens capsule and the basal lamina of the ciliary epithelium. 5, Hyaluronidase only slightly changed the lens capsule. 6. The vitreous fibers were digested by collagenase, but not by alpha-chymotrypsin or hyaluronidase. Thes results together with the review of recent literature indicate that the zonular fiber has a nature close to that of the microfibril of elastic fiber.


The influence of diazepam on the dosage of muscel-relaxants during anaesthesia. The literature on anaesthesiological studies performed to investigate a possible sparing effect of diazepam on the dosage of muscle-relaxants during anaesthesia is reviewed. Despite some conflicting results it can be concluded that diazepam exerts such an effect, thus allowing between 8 and 30% of the amounts of muscle-relaxants, depending on the type of anesthesia used, to be spared. This conclusion is also supported by the results of pharmacological studies in animals, using the head drop method. The mechanism of this effect is discussed on the basis of the relevant published pharmacological evidence from "in vitro" studies and from both animal and human "in vivo" studies. From these reports it is concluded that diazepam, at therapeutic doses, does not act on neuromuscular transmission and that this sparing effect does not result from direct pharmacological interaction with neuromuscular blocking agents. Some studies indicate a minimal action of the product on the skeletal muscle fibre, but this also is not related to the sparing effect. It is furthermore concluded that his sparing effect of diazepam on the dosage of muscle-relaxants during anesthesia is due to a summation of muscle-relaxant actions exerted at different levels of the nervous system, those of diazepam being mainly exerted at the spinal and brainstem level and consisting in inhibition of polysynaptic reflex activity, enhancement of presynaptic inhibition, and depression of gamma-motoneuron activity.


Actions of a new muscle relaxant (AH8165) on neuromuscular transmission. The effects of a new muscle relaxant, AH8165, on miniature endplate potential (MEPP) amplitude and frequency, endplate sensitivity to acetylcholine, and muscle twitch tension were studied in vitro in the frog sartorius muscle. Nerve terminal effects were studied in vivo in the cat soleus muscle and its ventral root fibers. AH8165 stimulates the nerve terminal, as evidenced by increased MEPP frequency and the appearance of post-drug repetitive activity. In the same concentration range at which MEPP frequency is increased, MEPP amplitude, endplate sensitivity to acetylcholine, and twitch tension are decreased. This suggests that AH8165 produces muscle relaxation by blocking postsynaptic cholinergic receptors. (Key works: Neuromuscular relaxants, AH8165.).


Use of a rapid brain-sampling technique in a physiologic preparation: effects of morphine, ketamine, and halothane on tissue energy intermediates. A new method of rapid sampling of brain tissue, "freeze-blowing," has been used to compare the neurochemistry of the brain during anesthesia with that in the awake state. The method avoids anoxia associated with the sampling process. Physiologic variables, including body temperature, blood-gas tensions and blood pressure, were carefully monitored and controlled in the experimental animals. None of the agents tested (halothane, morphine, and ketamine) reduced the brain tissue high-energy phosphate reserved. All three drugs doubled glucose levels. Morphine lowered both lactate and the lactate/pyruvate ratio. Uniformly, the three anesthetic agents led to twofold increases of brain cyclic 3'-5' adenosine monophosphate concentrations. These changes suggest a possible role for cyclic nucleotides in central neurotransmission.


The effect of dimethyl sulfoxide on the neuronal excitability and cholinergic transmission in Aplysia ganglion cells. The effects of DMSO on single nerve cells of Aplysia were investigated by various electrophysiological methods. 1. Although the increase in permeability of the biological membranes produced by DMSO has been well documented, we found that DMSO at concentration of less than 20% actually decreases the permeability of the neuronal membranes, probably toward potassium and chloride ions. This change in ionic permeability reversibly depolarizes the resting membranes and makes the neurons more excitable. 2. The falling phase of the spike is prolonged by 8% DMSO, because it blocks the active increase in potassium permeability. This change suppresses the high frequency discharge, because the refractory period of each spike is increased. Neither the rising phase nor the absolute firing level is appreciably altered by DMSO when these are examined after the depolarization is cancelled. 3. Dilute DMSO (less than 1%) facilitates cholinergic trasmission because it blocks ACh-esterase activity. DMSO at concentrations of more than 10% ultimately blocks cholinergic transmission entirely, because it also depresses cholinoceptive receptor activity (probably by allosteric interaction). The action of 1-10% DMSO is complicated, because both facilitatory and inhibitory effects take place at the same time. 4. In cholinergic synapses, excitatory transmission is more susceptible to DMSO than inhibitory transmission. This is because the activity of the excitatory receptor is blocked more readily than that of the inhibitory receptor. The depressing effects of DMSO are not specific to the cholinergic system, however, since the activities of GABA and glutamate receptors are similarly depressed.


Specific modifications of the Na+,K+-dependent adenosine triphosphatase by dimethyl sulfoxide. DMSO inhibits the Na+, K+-ATPase, but stimulates the associated K+-phosphatase activity. For the ATPase, DMSO acts as an uncompetitive inhibitor toward both ATP and Na+, whereas it increases the K0.5 for K+. From measurements of the dissociation constant (Km) of these ions in the ligand states that correspond to the ATPase reaction, it can be shown that DMSO has little effect on the affinity for Na+, but decreases the affinity for K+ of the enzyme-phosphate intermediate (the form that has the highest affinity for K+). By contrast, DMSO decreases the Km for the phosphatase substrate (nitrophenyl phosphate) without affecting the Vmax. Moreover, DMSO decreases the K0.5 for K+ and also the Kd for K+ in the ligand states that correspond to the phosphatase reaction (which have only a moderate affinity for K+, since the acyl phosphate intermediate is absent in this pathway). These data may be incorporated into a reaction mechanism for the Na+, K+-ATPase. Initially the enzyme is phosphorylated to form an acyl phosphate intermediate, in steps that require Na+ and Mg-2+. At this stage the affinity of K+ is markedly increased (from the moderate affinity seen in the "free" enzyme and the phosphatase reaction). When K+ is bound, the phosphate group is transferred to the hydrolytic site where P-i is ultimately released. DMSO acts at the point at which the acyl phosphate group or the phosphatase substrate enters the hydrolytic site, inhibiting one and facilitating the other. At this stage the affinity for K+ is also changing, and DMSO apparently selects an enyme conformation of intermediate affinity. Ion transport may occur by a gate mechanism in an overall system that operates on a half-of-the-sites active enzyme pattern in which ATP hydrolysis may alternate between the dimeric subunits of the enzyme.


Thymosin-induced differentiation of murine thymocytes in allogeneic mixed lymphocyte cultures. Calf thymosin is shown to enhance the one-way MLR of CBA thymocytes cultured with allogeneic mitomycin-C- treated C57BL/J6 spleen cells. Thymosin does not enhance the one-way MLR of CBA thymocytes cultured with syngeneic mitomucin-C-treated spleen cells. Based on this finding we present a relatively simple, rapid and quantitative in vitro microculture hioassay for inducers of T-cell differentiation and propose that thymosin treatment, when accompanied by antigen presentation, induces the two-step maturational sequence of pre-T yields T1 yields T2.


The isolation of thymopoietin (thymin).

The isolation of thymopoietin (thymin). The isolation from bovine thymus of two closely related polypeptides, thymopoietin I and II, is described. These are considered to be thymic hormones, which physiologically induce the differentation of prothymocytes to thymocytes within the thymus. Ths isolation of the thymopoietins was monitored not by their differentiative effects, but by a presumably secondary effect on neuromuscular transmission. This was discerned in experimental studies related to the human disease myasthenia gravis in which it was suggested that autoimmune thymitis was regularly present. In an animal model, experimental autoimmune thymitis, the thymic disease was shown to result in the release of a substance that depressed neuromuscular transmission and this substance was shown to be also secreted in small amounts by the normal thymus. A bioassay was developed, this being the delayed appearance of neuromuscular impairment after in vivo injection of the active material, and this bioassay was used to monitor the fractionation of thymus extracts and isolate thymopoietin. Pure thymopoietin was active at subnanogram concentrations, both in producing its effect on neuromuscualr transmission and in inducing the differentiation of prothymocytes to thymocytes. This potency of activity of the purified polypeptide, as well as its specificity in inducing the differentiation of T-cells and not B-cells, support the consideration that thympoietin is a physiological inducing hormone produced by the thymus. This is further supported by the evidence that thymopoietin is only produced in the thymus: neuromuscular blocking effects are not present in extracts of other tissues and immunofluorescent localization of thymopoietin shows it to be present only in thymic epithelial cells.


Regulation of theta-antigen expression by agents altering cyclic AMP level and by thymic factor. Thymic factor, cyclic AMP, and products increasing its cellular level, such as Prostaglandin E1, induce the appearance of the theta-antigen on T-cell precursors whether assessed by a rossette-inhibition assay or a cytotoxic assay after cell fractionation on BSA discontinuous gradiet. Synergism has been demonstrated between cyclic AMPT and TF for that effect. Conversely, decrease of theta expression has been obtained by altering cyclic AMP level in theta-positive cells either increasing it by dibutyryl cAMP treatment or decreasing it by indomethacin treatment. Finally, these data suggest the involvement of cyclic AMP in the regulation of theta expression under thymic hormone control.


Influence of the thymus-corticothropin-growth hormone interaction on the rejection of skin allografts in the rat. Skin allograft rejection was noticeably delayed in rats following hypophysectomy. Daily injections of hypophyseal growth hormone restored the normal reaction. Additional thymectomy had no influence on the rejection of hypophysectomized rats. In these animals growth hormone by itself had no significant influence on the graft rejection. It only restored a normal reaction when given together with thymic hormone. Corticotropin injections accelerated the allograft rejection. On this action of corticotropin thymic hormone has shown a significant inhibitory influence. The thymus was thus proved to be a synergist to growth hormone and an antagonist to corticotropin. Previous observations made with usual endocrinological test- are thus confirmed with an immunological test.


Stimulation of cAMP levels and modulation of antibody formation in mice immunized with cholera toxin. Injection of mice with 1.0 mu g of a purified exotoxin derived from Vitro cholerae together with a challenge injection of sheep erythrocytes (SRBC)P OR E. coli LPS markedly influenced the immune response to these antigens. Simultaneous injection of the toxin with antigen resulted in a delayed appearance of antibody-forming cells during the first few days after immunization, followed by a marked enhancement of the peak numbers of antibody-forming cells. In the case of the immune response to SRBC, both 19S and 7S plaque-forming cells (PFC) were enhanced on the peak day of response after simulataneous immunization of toxin-injected mice. The secondary immune response to SRBC was also similarly affected when cholera toxin was given along with a second injection of erythrocytes: i.e. a delay in appearance of the first antibody-forming cells followed by a marked enhancement of the peak 19S and 7S PFC response. Injection of cholera toxin 103 days prior to SRBC or LPS was immunosuppressive. The effect of cholera toxin on the level of splenic cyclic AMP appeared related to the effects on antibody formation.


The latero-medial inferior pontine syndrome. A case is presented with a pontine vascular lesion involving the V, VI, VII, and VIII cranial nerves. The diagnostic evaluation included audiometry and electronystagmography as well as arteriography, pneumoencephalography, and brain scan. Total deafness developed in the involved ear and was present for seven weeks before complete recovery occurred. Electronystagmographic findings consistent with a brain stem lesion also resolved following recovery of the patient. Audiometry and electronystagmography are described and documented for the first time in a report of a patient with a latero-medial inferior pontine syndrome. A discussion of some of the unique features of the brain stem vasculature and its relationship to clinical syndromes involving the brain stem is given.


Value of special auditory tests in the latero-medial inferior pontine syndrome. Results from a special battery of auditory tests are used to document, for the first time, recovery from a total unilateral hearing loss occurring as part of a latero-medial inferior pontine syndrome. Although sensitivity for pure tones and speech was regained within two months of onset, definite retrocochlear signs persisted for nine months. The tests which best demonstrated the lingering retrocochlear abnormality were binaural masking level difference procedures for 500 Hz pure tones and speech and examination of the amplitude of the acoustic reflex over a ten second time period. The relative sensitivity of other speech and pure tone tasks in detecting the retrocochlear problem is detailed.


Basal cell carcinomas and basal cell carcinoma-like changes overlying dermatofibromas. Three patients had pigmented nodular basal cell carcinomas overlying dermatofibromas and 19 patients had basal cell carcinoma-like changes. We agree with the view generally held that acanthosis, pseudoepitheliomatous hyperplasia, basal cell buddings, and hair follicle-like proliferations of basal cells represent reactive changes of a spectrum of epidermal alterations overlying dermatofibromas. Contrary to the general opinion, however, we believe that basal cell carcinoma-like changes, superficial multicentric basal cell carcinomas, and nodular basal cell carcinomas represent neoplastic changes of such a spectrum.


A syndrome of trichoepitheliomas, milia, and cylindromas. Three consecutive generations of a family have had members affected with trichoepitheliomas and milia. One member, the proband, also had cylindromas. The pattern of inheritance was a dominant one, with only the male members affected. Histopathologically, the trichoepitheliomas and milia contained keratinizing cysts with laminated centers, peripheral basaloid cells, and a thin granular layer. Fibroblastic stroma and fronding of basaloid cells were also seen in both tumors. The trichoepitheliomas and milia differed mainly in the greater proportion of keratinized cells found in the milia. Clinically, the milia were found only in areas of vellus hairs.


EB-virus-specific IgM and IgG antibodies in first-degree relatives of children with acute lymphoblastic leukaemia. EB-virus-specific IgM and IgG antibodies (to virus capsid and soluble complement fixing antigens) were estimated in sera from mothers and sibs of children with acute lymphoblastic leukaemia, from patients with infectious mononucleosis, and from control induviduals. IgM antibodies were present in 12 of 16 mothers and 3 of 4 sibs of children with acute lymphoblastic leukaemia. They were also present in 14 of 16 patients with infectious mononucleosis, but in only 1 of 12 control individuals.


Sexual precocity associated with a hypothalamic tumour. Effects of sex hormone therapy. A case of sexual precocity of unusually early onset and associated with a hypothalamic tumour is described. The effects of ethinyl oestradiol and medroxyprogesterone acetate treatment have been monitored by plasma testosterone, FSH, and LH estimations. The results suggested a partial suppression of the tumour's influence, the mechanism of which is discussed.


Bone changes in congenital cytomegalic inclusion disease. We report the case of a female infant born with cytomegalic inclusion disease, in whom bone changes were present in x-rays of the tibiae and femora. She was the mother's first child and was born 8 days postmature after a normal delivery; birthweight 2.52 kg. Many infarcts were seen in the placenta. The mother had a feverish illness at the 7th month of gestation, diagnosed by family doctor as influenza. No drugs had been given.


Separation of human epidermal cells from fibroblasts in primary skin culture. To obtain pure culture of epidermal cells from small human biopsies, two different techniques were tested and compared, i.e. separation of epidermis from corium before cultivation by trypsin and suction, and after cultivation by trypsin and collagenase. The most active growth of epidermal cells was obtained by the third technique, since short-term trypsin treatment released only fibroblasts from the culture. Crude collagenase (type I) was less effective than trypsin. Collagenase type II, III and IV had no effect on fibroblast release. Neither trypsin nor collagenases dispersed epidermal cells.


Metastatic cystosarcoma phylloides in an adolescent girl: an unusually malignant tumor. The first case of metastatic cystosarcoma phylloides in an adolescent is reported. This case also represents the first reported response of this tumor to chemotherapy and irradiation. This tumor showed an unusually rapid rate of cell division probably making it more susceptible to these agents. The importance of an early diagnosis and definitive excision is stressed.


Factors determining successful liver preservation for transplantation. Auxiliary liver allotransplants will survive for relatively long periods of time after 24 hour hypothermic (10-12C), pulsatile perfusion. The best perfusate was a silica gel fraction of dog plasma with added potassium chloride gel made hyperosmolar with glucose. Further improvement could be achieved with added allopurinol and methylprednisolone. Nonpulsatile flow or lower temperatures were less effective preservation techniques.


Treatment of intra-abdominal and abdominal wall desmoid tumors with drugs that affect the metabolism of cyclic 3',5'-adenosine monophosphate. A female patient with Gardner's syndrome was treated with delta1-testololactone (200 mg daily) because of growth of a large desmoid tumor in the pelvis and lower abdomen and a tumor in a scar from a previous laparotomy. There was also pain and swelling of the left leg. An immediate effect of the drug therapy was complete relief of pain followed shortly thereafter by disappearance of the edema of the leg. After two months, the numerous sebaceous cysts were less prominent. The gross measurements of the diameter of the pelvic and lower abdominal tumor clearly demonstrated tumor shrinkage following therapy. Small polyps scattered over the rectal mucosa and numerous osteomata were not demonstrably affected. After one year of treatment with delta1-testololactone, a laparotomy for partial small bowel obstruction was necessary. Obstruction was caused by the involvement of small bowel mesentery and the bowel itself in a contracted residuum of dense fibrous tissue. Substitution of theophylline and chlorothiazide for the testololactone in Januray 1974 was followed by further diminution of the measurable abdominal and pelvic desmoids. All of these compounds synergize the action of 3',5'-adenosine monophosphate and at least the latter two may function by inhibiting the action of 3',5'-adenosine monophosphate diesterase.


Cancer of the bile ducts associated with ulcerative colitis. Thirteen patients with bile duct cancer (excluding gallbladder) and associated chronic ulcerative colitis (CUC) were seen at the Mayo Clinic from 1935 through 1973. Most patients had initial symptoms of severe diarrhea and bleeding, followed by a pattern of mild-to-moderate disease with exacerbations and remissions. Three patients had especially severe symptoms and underwent total colectomy (1 patient) or proctocolectomy (2 patients) an average of 15.7 years from onset of CUC symptoms. Anorexia, followed rapidly by the development of progressive jaundice (or a sudden deterioration when liver disease was already present), marked the onset of symptoms of bile duct cancer in the 13 patients. The overall mean duration from onset of CUC to development of symptoms of bile duct cancer was 19 years. The patients in whom colitis was managed by proctocolectomy or total abdominal colectomy developed symptoms of bile duct cancer an average of 9.4 years after colectomy. When cancer was diagnosed, the tumor had spread beyond the bile ducts in 10 patients. The tumors were difficult to identify and often infiltrated the hepatic hilus. The present series and review of the literature suggest that the relationship between CUC and bile duct cancer is more than a chance occurrence. The carcinoma has an onset approximately 3 decades earlier than does carcinoma of the bile ducts without CUC. Surgical removal of the diseased colon and mode of medical management of the unresected colon have no relationship to the subsequent development of carcinoma of the bile ducts; neither does the extent or severity of the colonic disease. The prognosis of carcinoma of the bile ducts unfortunately continues to be dismal.


Occult adrenal insufficiency in surgical patients. Eight patients admitted to a University hospital with acute surgical problems and related adrenal insufficiency were reviewed and three are presented in detail. Surgical stress and continued sepsis played major roles in the lack of responsiveness to usual modes of therapy until the adrenal insufficiency was corrected. The patients fell into three major clinical categories of adrenal insufficiency. Chronic illness and sepsis are shown to affect steroid production and metabolism, as well as adrenal responsiveness to ACTH. Pharmacologic amounts of steroids are often needed in patients with shock, gram negative sepsis and prolonged illnesses, even if normal or elevated serum cortisols are present. Therapeutic trials of cortisol administration are shown to be confusing when not accompanied by easily performed diagnostic tests of adrenal function. It is emphasized that a pretreatment serum cortisol should be obtained whenever possible. The evaluation of adrenal function is of lifelong importance to the patient.


Herpetic tracheobronchitis. Cytologic and virologic detection. Tracheobronchitis due to herpes simplex virus is a well-recognized finding in cases of burns, debilitation, or immunosuppression. Nearly all reported cases have been diagnosed at necropsy despite the possibility for clinical detection of such infections by exfoliative cytological studies, virus isolation and identification, or both. The present report details the cytologic and virologic diagnosis of herpetic tracheobronchitis in a patient with carcinoma of the lung and alcoholic fatty liver. Respiratory cells with herpetic infection cytologically showed less tendency to multinucleation than the characteristic herpes-infected cells of squamous epithelium, which may be a source of diagnostic confusion.


Recovery from encephalomyelitis caused by Herpesvirus simiae. Report of a case. A 29-year-old monkey handler developed an acute encephalomyelitis with neuromuscular dysfunction that progressed to respiratory arrest on the 18th day of illness. Thereafter, with supportive care, the patient's condition improved steadily. The titer of neutralizing antibodies to H simiae rose from 1:4 (eighth day of illness) to 1:512 (47th day of illness). Apparently the fifth known survivor of H simiae (herpesvirus B) encephalomyelitis, this patient is also remarkable because of virtually complete recovery, apparently the second documented instance of a good outcome.


A review of REM sleep deprivation.

A review of REM sleep deprivation. Studies on the behavioral consequences of rapid eye movement (REM) sleep deprivation in animals and humans are critically reviewed. In animals, converging evidence--some reasonably well controlled--indicates that REM sleep deprivation probably heightens central neural excitability and increased motivational behavior, but has nuclear or inconclusive effects on learning. In humans, evidence indicates that REM sleep deprivation is not dream deprivation and is not harmful to schizophrenic, depressed, or healthy subjects. Controversy continues about whether or not (some) schizophrenic patients respond abnormally to REM sleep deprivation by having no REM rebound. Controlled but unconfirmed work indicates that that endogenous, but not reactive, depressive patients are improved by REM sleep deprivation, a finding consistent with the animal behavioral consequences of the procedure and with the unique REM-depriving properties of efficacious antidepressant drugs.


REM sleep reduction effects on depression syndromes. Thirty-four endogenous and 18 reactive, depressed patients (hospitalized and nonschizophrenic) were treated in a double-blind, crossover study of the hypothesis that rapid eye movement (REM) sleep reduction (by awakenings) relieves depression. In the endogenous group-but not in the reactive group-subjects deprived of REM sleep for three weeks improved significantly more than control subjects awakened from non-REM sleep. Therapeutic efficacy of REM sleep reduction appeared similar to reported efficacy of imipramine hydrochloride treatment of depression. Eight of nine endogenous patients, unimproved by REM sleep deprivation, did not improve with imipramine. Results suggested (1) that substantial REM sleep reduction has antidepressant activity, and (2) since imipramine and other drug antidepressants reduce REM sleep much more so than nonantidepressant drugs, that an antidepressant "mechanism" of drugs resides in their capacity to substantially reduce REM sleep.


A neuroendocrine predisposition for homosexuality in men. In male rats, androgen deficiency during a critical hypothalamic organizational period was shown to give rise to a predominantly female-differentiated brain, homosexual behavior, and demonstration of a positive estrogen feedback effect. A positive estrogen feedback effect was also induced in intact homosexual men in contrast to intact heterosexual and bisexual men. Thus in 21 homosexual men an intravenous injection of 20 mg Presomen (Premarin) produced a significant decrease of serum LH levels followed by an increase above initial LH values. In 20 heterosexual and in five bisexual men, by contrast, intravenous estrogen administration, while producing a significant decrease of the serum LH level, was not followed by an increase above the initial LH values. Using a radioimmunoassay, plasma testosterone levels and 24-hr urinary excretions of unconjugated testosterone of adult homosexual men were found to be in the normal range as observed in heterosexual men. This finding suggests that homosexual men possess a predominantly female-differentiated brain which may be activated to homosexual behavior by normal or approximately normal androgen levels in adulthood.


Further evaluation of total pancreatectomy. The results of 64 total pancreatectomies performed at the Mayo Clinic between 1942 and 1973 have been reviewed. Improvement in recent results as compared to the previously reported series is probably related to improvements in operative techniques and in selection of patients with more favorable malignant lesions. The recent results are more favorable than those reported for a group of comparable patients undergoing standard Whipple procedures for malignant lesions at this institution. Diabetes was easily managed in 76% of patients. Hypoglycemia due to insulin sensitivity is the major problem, and ketoacidosis seldom occurs. Recognition of this has resulted in better management of diabetes with relatively small doses of insulin. The sequels of long-standing diabetes do not appear to be a problem. Gastrointestinal tract hemorrhage has been greatly decreased through the use of more extensive gastric resection.


[A change in the ultrastructure of frog spinal cord and motor neuron synapses during prolonged activation]. Motoneuron axosomatic synapses (AS) were shown to be larger than axodendritic (AD) synapses under anaesthesia. A 10 minute-long electrical stimulation both by low and high frequency caused enlargement of AS and AD synapses, it being the more pronounced the higher was the activation frequency. In all cases changes the AS synapses were more marked than those of AD synapses. On the bases of morphological and physiological data it was concluded that the changes of the synapse dimentions did not affect the synaptic transmission.


[A malignant variant of Abrikosov's tumor]. Three observations on granular-cell Abrikosov's tumours with locally destructive growth and polymorphism of cells (in 2 male and 1 female patients) located in soft tissues of a hand finger, under the skin of teh upper lip, and in soft tissues of the lumbar region are described.  A critical survey of the literature is given.


[Fibroblast-fibroclast: The ultrastructural mechanisms of resorption of collagen fibers in involution of the connective tissue]. Using various experimental models (postpartum involution of the uterus, resolution of a subcutaneous scar which replaced the collagen implant and connective-tissue capsule following the removal of a foreign body) and electron-microscopy and histochemical methods of investigation intra- and extracellular resorption of th  e collagenous fibres was extablished.  The main role in this process was played by fibroblasts which uncer these conditions could funciton as fibroclasts phagocyting and idgesting in its cytoplasma collagenous fibrillas with the help of lysosomal enzymes.  Desintegration of fibroclasts, enriching the medium with these enzymes, stimulated the extracellular lysis of the collagenous fibrillas.  This lysis was particularly intensive in the presence of an inflammatory process, and even could take place before the intracellular resorption.  In macrophages no intracellular lysis was observed, however, they could phagocyte denaturated collagen, deprived of structural orgainzation.


Hydrocephalus produced by the 6/94 virus; A parainfluenza type 1 isolate from multiple sclerosis brain tissue. The 6/94 virus, parainfluenza type 1 isolate from multiple sclerosis brain tissue, produced hydrocephalus in newborn Syrian hamsters. All animals developed clinical disease and died within a week. Ependymal cells lining the aqueduct of Sylvius became necrotic and fused, resulting in obstructive hydrocephalus. The 6/94 virus antigen was seen in ependyma and meninges. Paramyxovirus nucleocapsids were seen within cytoplasm of ependymal cells. Virus was recovered from hamster brains for only two days. Infectious virus could be recovered from brains grown in vitro as explants for 21 days. No evidence of rising hemagglutination-inhibiting antibody was noted for up to one month after infection. Intraperitoneal or subcutaneous injection of 6/94 virus did not produce hydrocephalus. HA2 virus and the temperature sensitive mutant of HA2 virus failed to produce hydrocephalus, while Sendai virus caused lesions similar to those of 6/94 virus.


Atypical myopathy with myofibrillar aggregates. An autopsy of a 23-year-old woman with progressive muscular weakness and wasting showed a unique muscle abnormality with segmental involvement of individual fibers by peculiar inclusions. Electron microscopically, these inclusions resembled cytoplasmic bodies, being formed of two concentric zones of filamentous materials. They seemed to arise from filaments of myofibrils that were fragmented and highly disorganized in affected areas.


Lipoproteins in atherosclerotic lesions. Localization by immunofluorescence of apo-low density lipoproteins in human atherosclerotic arteries from normal and hyperlipoproteinemics. Using immunofluorescence techniques, apoLDL, the major protein component of human plasma low and very low density lipoproteins was localized in athersclerotic lesions from the aorta, coronary, and carotid arteries in human normolipoproteinemic, type II, and type IV hyperlipoproteinemics. Although no apoLDL was detected in uninvolved arteries, atheromas from advanced lesions as well as certain collagen and elastic fibers from both fatty streaks and advanced lesions were positive for apoLDL. This localization pattern was the same in each lipemic group, except for the presence of apoLDL in some intimal cells from type II cases. However, extensiveness of apoLDL localization was greatest in the type II group, but equal in the normolipemic and type II group. This may be explained by the high concentration gradient between plasma and tissue for apoLDL in the type II group.


Collagen metabolism in rat lungs during chronic intermittent exposure to oxygen. Collagen metabolism was studied in rat lungs during prolonged intermittent exposure to oxygen. After an initial of 2-d exposure to oxygen, the rats alternately breathed air and oxygen in 24-h periods up to 30 d. Lung histology showed perivascular edema as an early sign of oxygen toxicity. Increased capillary permeability was followed by a marked perivascular leukocytosis that was prominent between the 5th and 11th d of the treatment. During the further course of the experiment, the inflammatory response gradually diminished, and finally a network of collagen fibres filled the perivascular areas. The inflammatory process elevated the amounts of DNA and RNA in lung tissue, and the ratio of RNA to DNA was maximal on the 5th day. Studies on metabolism of lung collagen with intraperitoneally injected 3H-proline as label showed that, after the initial inflammatory reaction, both synthesis and degration of collagen were increased in oxygen-treated lungs. The increase in synthesis exceeded the rise in degradation and, therefore, collagen was accumulated. The activation of lung collagen synthesis coincided in time with a rise in the activity of lung protocollagen proline hydroxylase.


Histologic types and possible initial stages in early gastric carcinoma. Among 2003 gastric specimens from 301 patients, diagnosis for carcinoma was made in 45 cases.  Examination of resection preparations revealed 36 cases of deep invasive stomach carcinoma and 9 cases of early gastric cancer confined to mucosa and/or submucosa.  Carcinomatous proliferations limited to mucosa or submucosa are classified in three histologic types: intestinal (adeno), mucocellular (signet ring cell), and anaplastic (solid) type of early gastric cancer.  Mixed types have been found combining the first and the second, or the second and the third type.  One case presented a mixture of all three types.  Possible precursor or initial stages of all three types were found in further 31 biopsies.  Some of them were glandular lesions in superficial parts of the mucosa; this kind has been described previously as possible preneoplastic stage of the intestinal type of early gastric cancer.  "Signet ring cell drippings" from lower parts of tubule necks were recorded as an initial form of the signet ring cell type.  The process is interpreted as detaching of isolated signet ring cells from a gland neck zone in progressing atypical transformation.  An early neoplastic stage of the anaplastic (solid) type of early gastric cancer is identified in the "gland neck dysplasia" located exclusively in the antrum between surface mucosa and antrum glands.  This lesion appears rich in cells and stretched like a broad ribbon.  Early gastric cancer of this third type will arise in the very same location.  Conclusions from formal histogenesis suggest that the signet ring cell type and the anaplastic (solid) type of early gastric cancer might start in the lower part of tubule necks.  In consequence, the neck region of gastric glands could be the critical field of malignant transformation in the gastric mucosa.  Long-term follow-up studies will be needed to verify these observations and their interpretation.