Factors affecting the solubility of calcium pyrophosphate dihydrate crystals. The solubility of triclinic calcium pyrophosphate dihydrate (CPPD) crystals was measured under varying conditions using 45Ca-labeled crystals, expressing solubility as micromoles per liter of 45Ca in solution. In a 0.1-M Tris-HC1 buffer pH 7.4, the solubility of accurately sized CPPD crystals (37-20mum) was 60muM with maximal solubility being attained after about 8 h incubation at 37degreeC. Reduction in crystal size, decrease in pH, increase in ionic strength, Mg++, citrate, and albumin all increased solubility. The most marked effects on solubility occurred when changing the calcium concentration or by enzymatic hydrolysis of inoganic pyrophosphate to orthophosphate. It was found that decreasing the ionized calcium level below 5 mg/100 ml resulted in a progressive enhancement of solubility. The observed solubility-enhancing effects of albumin could be explained solely on its calcium-binding ability and thereby, altered ionized calcium level. Diffusible calcium in synovial fluid was only 40% of the total calcium concentration, which means most joint fluids are normally near the critical concentration of 5 mg/100 ml of ionized calcium, below which solubility is enhanced. During surgery, especially parathyroidectomy, calcium levels fall, favoring dissolution of CPPD crystals. We speculate that the slight decrease in crystal size during dissolution frees them from their cartilaginous mold, resulting in a dose-dependent inflammatory reaction as they are "shed" into the joint space. Crystal shedding may be reinforced by the modest fall in joint fluid pH accompanying the inflammatory response.


Evidence from rats that morphine tolerance is a learned response. It is proposed that the direct analgesic effect of morphine becomes attenuated over the course of successive administrations of the narcotic by a conditioned, compensatory, hyperalgesic response elicited by the administration procedure, the net result being analgesic tolerance. Using the "hot plate" analgesia assessment situation with rats, this conditioning view of tolerance is supported by several findings: (a) It is necessary to have reliable environmental cues predicting the systemic effects of morphine if tolerance is to be observed, (b) a hyperalgesic conditioned response may be observed in morphine-tolerant subjects when drug administration cues are followed by a placebo, and (c) merely by repeatedly presenting environmental cues previously associated with morphine (but now presented with a placebo), morphine tolerance can be extinguished.


Murexide for determination of free and protein-bound calcium in model systems. The determination with murexide of free and protein-bound calcium in model systems of known composition, ionic strength, and pH was investigated. The spectra of calcium murexide in the presence of varying amounts of calcium ions indicated that the absorption maximum fo calcium murexide complex occurs at 480 nm while that of murexide ion is at 520 nm. The absorbance at 509 nm is independent of calcium ion concentration and, therefore, could be used to measure the total dye. The spectra are pH dependent but constant in the range 6.5 to 7.0. The apparent dissociation constant of calcium murexide is dependent upon ionic environment, ionic strength, and free calcium ion concentration. The relationship between the apparent dissociation constant and free calcium concentration was established. Whole casein had no effect on the absorption spectra of calcium murexide and no affinity for calcium murexide complex or murexide ion. Beta-casein, at the concentrations employed, did not influence the dissociation fo calcium murexide. At pH 7.0, ionic strength .1, and 2 C, Beta-casein bound calcium as if there were 8.65 binding sites per molecule, each of pK 2.23, corresponding to an intrinsic association constant of 168.9 liters per mole.


Lorazepam compared with pentobarbital for nighttime sedation. Lorazepam (0.5, 1, 2, and 4 mg) was compared with pentobarbital (60 and 180 mg) for its effect on sleep in "hospital insomnia." Subjective-response data were collected by research nurses. Lorazepam was found to be a potent nighttime sedative: 1 to 1.25 mg of lorazepam is equivalent to 100 mg sodium pentobarbital for measures of sleep quality and duration. At this dose level it is less effective than 100 mg of pentobarbital as a sleep inducer. Studies at higher doses (up to 4 mg) indicate that lorazepam has a wide therapeutic index.


Effect of trace elements on dissolution of hydroxyapatite by cariogenic streptococci. The effect of low levels of strontium, boron, lithium, molybdenum, and fluorine, alone and in combination, on hydroxyapatite solubility, bacterial growth, and acid production in five antigenic types of Streptococcus mutans was investigated. Pour plates containing synthetic hydroxyapatite were used to compare dissolution of hydroxyapatite. The colonies of the five antigenic types of S mutans produced zones of dissolution that were measured. Acid production and growth were studied in broth culture media. The results show that low levels of strontium and fluorine can significantly reduce demineralization of synthetic hydroxyapatite by S mutans in vitro.


[Use of isotopes in the diagnosis of malignant breast tumors]. The authors, with 67 Gallium, have obtained positive scintigraphy of the breast only in cases of carcinoma. Reliability of negative scintigraphy however is less good. Isotopic investigation of the bones is important in breast cancer and reveal early osseous metastasis.


Peroxisome development in the metanephric kidney of mouse. The relationship of enzymatic activity to organelle development and organelle number during differentiation of the metanephric kidney in the mouse was approached from several experimental directions. Biochemical analyses of marker enzymes for peroxisomes (catalase and D-amino acid oxidase), mitochondria (cytochrome oxidase) and lysosomes (acid phosphatase) were performed on kidneys at ages from 17 days prenatal to adult. These data were correlated with a morphometric analysis of populations of peroxisomes and mitochondria in differentiating cells of the proximal tubule. Postnatal development of the metanephric kidney was found to be accompanied by a rapid increase in both the specific activity of catalase and the number of peroxisomes per 100 mu2 in the proximal tubule during the first 4 weeks of postnatal growth. Elaboration of the endoplasmic reticulum (ER) was seen to parallel the increase in number of peroxisomes to which segments of ER were often in close apposition. Extensive interactions between segments of ER and peroxisomes were readily visible in 0.5-mu sections viewed in the high voltage electron microscope. In contrast to peroxisomes, neither mitochondria nor lysosomes followed a similar pattern of net organelle increase, suggesting that a defined population density of mitochondria and lysosomes may exist in the proximal tubule at birth, prior to complete development of the kidney.


Inhibitory effects of antihistamines and antiserotonins on the bone marrow reactions produced by Escherichia coli endotoxin in mice. The bone marrow reactions (that is, decrease of nucleated cell counts and increase of red blood cell counts) of mouse bone were observed 1 hr after injection of endotoxin and peaked after 18 hr. These reactions were significantly inhibited when diphenhydramine, promethazine (antihistamines), chlorpromazine (antiserotonin), or cyproheptadine (antihistamine and antiserotonin) was given 30 min before endotoxin. Such bone marrow reactions were also induced with histamine or serotonin and peaked 1 hr after administration. The histamine-induced changes were inhibited by prior treatment with diphenhydramine. These reactions were also produced by injection of a small amount of both histamine and serotonin, whereas no change was found when mice were given a single injection of a larger dose of histamine or serotonin. These results indicate that histamine and serotonin released in mice at the initial stage after endotoxin synergistically trigger the bone marrow reactions, which then continue in the presence of further mediators. Antihistamines and antiserotonins are considered to hinder the whole process of reactions produced by endotoxin.


On the interactions between pancreatic lipase and colipase and the substrate, and the importance of bile salts. The interactions between pancreatic lipase and colipase and the substrate and the effect of bile salts on these interactions have been investigated by the use of kinetic experiments and studies on the semiquantitative phase distribution of lipase and colipase activities. The results suggest that lipase binds to hydrophobic interfaces with partial irreversible inactivation. Bile salts in the range of micellar concentrations and above a pH of about 6.5 displace lipase from this binding, resulting in a reversible in activation. At pH values below about 6.5, lipase binds strongly to the substrate even in the presence of bile salt, and a low activity peak is seen around pH 5.5. This is the result of the binding of lipase to the "supersubstrate" and the activity of the catalytic site. In the presence of bile salt, colipase promotes the binding of lipase to the "supersubstrate" but not to other hydrophobic interfaces, and catalytic activity is reestablished. Kinetic data indicate that the binding between colipase and lipase in the presence of substrate is strong and occurs in an approximately stoichiometric relationship.


Effect of ionic strength and ionic composition of assay buffers on the interaction of thyroxine with plasma proteins. When plasma proteins are diluted with buffer the ionic strength and ionic composition of that buffer affects the interactions between thyroxine (T4) and its plasma protein-binding sites. Increases in phosphate, chloride or barbiturate ion concentration from 50 to 200 mmol/l caused a significant decrease in the affinity of plasma proteins for T4, and a concurrent increase in the concentration of unbound T4. These results cannot be completely accounted for by changes in ionic strength since at the same ionic strength different anions caused quantitatively different effects on unbound T4 concentration. The degree of depression of T4 binding by the three anions studied was in the order barbiturate greater than chloride greater than phosphate. The results of a systematic study on the composition of diluent buffer systems indicated that when a 50 mM-sodium phosphate-100 mM-NaCl buffer (pH 7-4) was used as a plasma diluent, there were unlikely to be gross changes in the T4-binding properties of plasma proteins with dilution.


The innervation of the salivary gland of the moth, Manduca sexta: evidence that dopamine is the transmitter. 1. Using the Falck-Hillarp histochemical technique for monoamines, evidence was found for the presence of a catecholamine in the salivary gland nerves of the moth, Manduca sexta. 2. The innervation was studied with the electron microscope. Only the fluid-secreting region of the gland is innervated and the nerve endings are characteristic of monoamine-containing terminals. 3. Using a sensitive enzymatic-isotopic assay for catecholamines, it was found that whole salivary glands contain 0.33 mug/g dopamine but no noradrenaline. 4. It seems likely that dopamine mediates fluid-secretion in the salivary gland of Manduca as it does a number of other arthropods.


An electrophysiological analysis of chemoreception in the sea anemone Tealia felina. 1. Electrophysiological techniques have been employed to examine the nature of the response observed in the ectodermal slow-conduction system (SSI) when dissolved food substances contact the column of Tealia felina. The response seems to consist entirely of sensory activity which may continue for periods of many minutes, provided that the stimulatory chemicals remain contacting the column. 2. The interval between each evoked pulse gradually increases as the sensory response progresses. This does not result from fatigue in the conduction system but involves a genuine process of sensory adaptation. This may occur over a period of several minutes, which is much longer than comparable adaptation in higher animals. 3. Physiological evidence suggests that the chemoreceptors involved are dispersed throughout the column ectoderm and are absent from the pedal disc, oral disc, tentacles and pharynx. 4. The basic role of the SSI in coordinating behavioural activity in sea anemones is reviewed. It is concluded that it functions primarily as a single, diffuse-conducting unit responsible for transmitting frequency-coded sensory information from ectodermal chemoreceptors to ectodermal (and perhaps endodermal) effectors.


The selective eosinophil chemotactic activity of histamine. Histamine diphosphate was shown to selectively attract human eosinophils from mixed granulocyte populations when over 20% eosinophils were used in a modified Boyden chamber chemotactic assay system. This effect of histamine is abolished by incubation with diamine oxidase (histaminase) and was generated by decarboxylation of L-histidine. A linear dose dependent increase in eosinophil migration was observed between 3 X 10(-7) M and 1.25 X 10(-6) M, while higher concentrations of histamine inhibited the migration of eosinophils. The attractant activity of histamine was not inhibited by H-1 or H-2 receptor antagonists, however, the inhibition of migration observed at higher histamine concentrations was reversed by metiamine, an H-2 receptor antagonist. The effects of histamine upon eosinophil migration were demonstrable using three different assays: (a) counting cells that had traversed 5-mum pore, 12-mum thick polycarbonate filters, (b) counting cells that had migrated various distances into a 3-mum pore, 145-mum cellulose nitrate filters, or (c) measuring the number of cells that had traversed an upper polycarbonate filter and migrated into a lower cellulose nitrate filter using 15Cr-labeled cells. The ability of histamine to enhance eosinophil migration was shown to be dependent upon the presence of a concentration gradient; histamine did not cause a dose-dependent increase in random motility. Furthermore, preincubation of the eosinophils with histamine deactivate the cells to further stimulation by histamine or by C5a. It is concluded that in low doses histamine is a chemoattractant for human eosinophils, while in higher doses histamine inhibits eosinophil migration. These observations may relate to the influx and localization of eosinophils in immediate hypersensitivity reactions.


Hemoglobins and hemocyanins: comparative aspects of structure and function. Comparative studies of protein structure and function can be quite interesting by themselves, and even more interesting when interpreted with respect to an animal's physiology. In the case of fish hemoglobins, some success in the latter has been achieved but there are still many unsolved problems. It appears that comparative physiology and biochemistry have entered an era where results from comparative studies can shed a great deal of light on biochemical mechanisms in general. The trout hemoglobin system is an example. Distinctive hemoglobins in this system are presently being used as high resolution probes of the ligand-binding mechanism. Characterization of the multiple, structurally distinct subunits of the 60S Limulus hemocyanin molecule may similarly aid in understanding its function. Our studies suggest the possibility of using Limulus hemocyanin and other hemocyanins as structural homologs and analogs of more complex macromolecular arrays. The rapid development of molecular structural data from X-ray crystallographers combined with the vast data of comparative physiology and biochemistry makes this one of the most exciting areas in present day science.


How do biological systems discriminate among physically similar ions? This paper reviews the history of understanding how biological systems can discriminate so strikingly among physically similar ions, especially alkali cations. Appreciation of qualitative regularities ("permitted sequences") and quantitative regularities ("selectivity isotherms") in ion selectivity grew first from studies of ion exchangers and glass electrodes, then of biological systems such as enzymes and cell membranes, and most recently of lipid bilayers doped with model pores and carriers. Discrimination of ions depends on both electrostatic and steric forces. "Black-box" studies on intact biological membranes have in some cases yielded molecular clues to the structure of the actual biological pores and carriers. Major current problems involve the extraction of these molecules; how to do it, what to do when it is achieved, and how (and if) it is relevant to the central problems of membrane function. Further advances are expected soon from studies of rate barriers within membranes, of voltage-dependent ("excitable") conducting channels, and of increasingly complex model systems and biological membranes.


Photoreceptor processes: some problems and perspectives. Visual photoreceptors from both vertebrates and invertebrates are characterized by extensive elaboration of membrane which contains visual pigment (rhodopsin). Visual pigments in all phyla examined are chemically similar: the chromophore is 11-cis retinaldehyde attached by an aldimine linkage (Schiff base) to a membrane protein, opsin. The effect of light is to isomerize the chromophore to the all-trans configuration. Beyond these fundamental similarities, several specific areas are discussed in which variations and differences appear. (1) Light causes vertebrate visual pigments to bleach, liberating the chromophore. Most invertebrate visual pigments do not bleach in the light, but instead form a thermally stable metarhodopsin, with the chromophore in the all-trans configuration still attached to the opsin. (2) In the disk membranes of vertebrate rod and cone outer segments, the rhodopsin molecules are oriented with their chromophores nearly coplanar with the disks. Within this plane, however, both rotational and translational diffusion are possible. In the microvillar membranes of arthropod and cephalopod rhabdoms, on the other hand, the situation is less clear. There is evidence for some preferential orientation of chromophores that implies restrictions on Brownian rotation. (3) In the outer segments of vertebrate receptors, absorption of light by rhodopsin causes the plasma membrane to hyperpolarize due to a decrease in sodium conductance, possibly mediated by calcium ions. In most invertebrate photoreceptors, light causes a depolarization due to an increase in conductance, principally to sodium ions. A subsequent entry of calcium causes a partial repolarization of the membrane, due to a decrease in sodium conductance. (4) For vertebrate receptors, log threshold is directly proportional to the fraction of rhodopsin bleached (Dowling-Rushton relationship). The proportionality constant varies in different preparations from less than four to more than 30, and the physical basis for the relationship is unknown. For invertebrates, by contrast, the dependence of sensitivity on rhodopsin concentration is much less dramatic and may well depend simply on the probability of quantum catch. (5) In most species, vertebrate and invertebrate, the accumulation of photoproduct probably has no effect on membrane conductance, but several possible exceptions exist. (6) Photoregeneration of rhodopsin from metarhodopsin is likely an important mechanism of recovery in certain arthropods such as diurnal insects, but dark mechanisms of recovery also exist in all phyla. In no single case are they adequately understood.


Hydrolysis of the hen egg vitelline membrane by cock sperm acrosin and other enzymes. A technique utilizing Pregnant Mare's Serum Gonadotropin and Human Chorionic Gonadotropin treatment of hens (Gallus domesticus), followed by manual ovulation of the excised follicles, was developed to obtain a large number of mature ova. The intact ova were used to test whether acrosin, partially purified from the spermatozoa of the cock (Gallus domesticus), partially purified rabbit testicular acrosin and commercial preparations of several hydrolytic enzymes could dissolve the inner vitelline membrane. Enzymes were applied to pieces of filter paper placed on the ovum. Cock acrosin and endopeptidases such as trypsin, chymotrypsin, collagenase and elastase hydrolyzed the membrane whereas exopeptidases such as leucine aminopeptidase and carboxypeptidase A did not. Phospholipase A, sulfatase, hyaluronidase, beta-glucuronidase and rabbit testicular acrosin also failed to hydrolyze the membrane. Cock acrosin hydrolysis of the ovum surface was inhibited by soybean trypsin inhibitor. The surface of the ovum over the germinal disc region was hydrolyzed more quickly by cock acrosin than the surface over other regions of the ovum. Acrosin from cock sperm caused the release of trichloroacetic acid soluble material absorbing at 280 nm from sonicated preparations of inner vitelline membranes. Hydrolysis was greatest at pH 8.0 and was inhibited by soybean trypsin inhibitor.


Some effects of low pH on chloride exchange in human red blood cells. In order to test the range of pH values over which the titratable carried model for inorganic anion exchange is valid, chloride self-exchange across human red blood cells was examined between pH 4.75 and 5.7 at 0 decrees c. It was found that chloride self-exchange flux had a minimum near pH 5 and increased again with further increase in hydrogen ion activity. The Arrhenius activation energy for chloride exchange was greatly reduced at low pH values. The chloride flux at pH 5.1 did not show the saturation kinetics reported at higher pH values but was proportional to the value of the chloride concentration squared. In addition, the extent of inhibition of chloride self-exchange flux by phloretin was reduced at low pH. Our interpretation of these findings is that the carrier-mediated flux becomes a progressively smaller fraction of the total flux at lower pH values and that a different transport mode requiring two chloride ions to form the permeant species and having a low specificity and temperature dependence becomes significant below pH5. A possible mechanism for this transport is that chloride crosses red cell membranes as dimers of HCl at these very low pH values.


Ionic properties of the acetylcholine receptor in cultured rat myotubes. The acetylcholine reversal potential (Er) of cultured rat myotubes is -3mV. When activated, the receptor is permeable to K+ and Na+, but not to Cl- ions. Measurement of Er in Tris+-substituted, Na-free medium also indicated a permeability to Tris+ ions. Unlike adult frog muscle the magnitude of Er was insensitive to change in external Ca++ (up to 30 mM) or to changes in external pH (between 6.4 and 8.9). The equivalent circuit equation describing the electrical circuit composed of two parallel ionic batteries (EK and ENa) and their respective conductances (gK and gNa), which has been generally useful in describing the Er of adult rat and frog muscle, could also be applied to rat myotubes when Er was measured over a wide range of external Na+ concentrations. The equivalent circuit equation could not be applied to myotubes bathed in media of different external K+ concentrations. In this case, the Er was more closely described by the Goldman constant field equation. Under certain circumstances, it is known that the receptor in adult rat and frog muscle can be induced to reversibly shift from behavior described by the equivalent circuit equation to that described by the Goldman equation. Attempts to similarly manipulate the responses of cultured rat myotubes were unsussessful. These trials included a reduction in temperature (15 degress C), partial alpha-bungarotoxin blodkade, and activation of responses with the cholinergic agonist, decamethonium.


Effect of cross-reinnervation on physiological parameters and on properties of myosin and sarcoplasmic reticulum of fast and slow muscles of the rabbit. Cross-reinnvervation of fast (extensor digitorum longus) and slow (soleus) twitch muscles of the rabbit showed essentially complete fast to slow and slow to fast conversion, respectively, 11-12 mo after surgery with respect to a number of physiological parameters including intrinsic shortening, velocity, and isometric twitch time to peak. There was pronounced bu incomplete biochemical conversion as judged by Ca2+ uptake by sarcoplasmic reticulum, myosin ATPase, alkali lability, and light chain complement. The question of trophic substances of neural origin is discussed in light of the fact that chronic stimulation for 15 wk of a fast muscle produces complete biochemical and physiological conversion to the slow type.


Chitin synthase in Mortierella vinacea: properties, cellular location and synthesis in growing cultures. Chitin synthase of Mortierella vinacea was present in the "microsomal' fraction (100 000 g precipitate), the 'cell-wall' fraction (2000 g precipitate) and the 'mitochondrial' fraction (10 000 g precipitate). The properties of the 'microsomal' enzyme were investigated. The pH optimum was between 5-8 and 6-2, and the temperature optimum was between 31 and 33 degrees C. The Km for UDP N-acetyl-D-glucosamine was 1.8 mM. The enzyme was stimulated by Mg2+ and a slight stimulation was also effected by N-acetyl-D-glucosamine. Soluble chitodextrins were inhibitory. A pH-dependent, heat-stable inhibitor of chitin synthase activity was present in the soluble cytoplasm from the mycelium. The effects of aeration and glucose concentration on enzyme production in growing cultures were also investigated; maximum specific activity of chitin synthase was associated with the cessation of exponential growth.


Protoplasts of Schizosaccharomyces pombe: an improved method for their preparation and the study of their guanine uptake. A new method is described for the efficient conversion of Schizosaccharomyces pombe cells into protoplasts. The following parameters of guanine uptake determined in whole cells were unchanged in protoplasts: Km value, requirement for an energy source, sensitivity to competitive inhibitors, pH optimum, as well as the typical variation of the initial velocity of uptake observed during the growth phase.


Manganese mutagenesis in yeast. A practical application of manganese for the induction of mitochondrial antibiotic-resistant mutations. When yeast cells were incubated for 4 to 8 h in yeast extract-peptone-glucose medium, pH 6, containing 8 mM-manganese, and then plated on selective media, there was a strong induction of antibiotic-resistant mutations. Indirect evidence suggests that practically all resistant mutants selected were of independent origin. The analysis of manganese-induced resistant mutants showed that most were extranuclear, while those tested showed recombination with known mitochondrial markers. Our results suggest that manganese can be considered as a mutagen which specifically induces mitochondrial mutations in Saccharomyces cerevisiae.


The properties and large-scale production of L-asparaginase from citrobacter. An intracellular L-asparaginase with antitumour activity was purified from a strain of Citrobacter. The optimum conditions for enzyme production by fermentation on scales up to 2700 l were investigated. Highest enzyme yield was obtained in corn-steep liquor medium (9-2%, W/V) at 37 degrees C. Oxygen limitation was not necessary for high enzyme yield. A total recovery of 4-3% from nucleic-acid-free extract and a 180-fold increase in specific activity were obtained after purificaiton. The specific activity of the purified preparation was 45 i.u./mg protein. The enzyme hydrolysed D-asparagine and L-glutamine at 7 and 5%, respectively, of its activity toward L-asparagine, but L-glutaminase activity could be demonstrated only at substrate concentrations above 5 mM. The Km values for L-asparagine and D-asparagine were 2-6 X 10(-5) and 1-4 X 10(-4) respectively. The anti-lymphoma activity of the enzyme was demonstrated with Gardner lymphosarcoma and was found only slightly less potent that Crasnitin, the most active asparaginase so far tested in this system.


Effect of inorganic phosphate on acridine inhibition and plasmid curing in Escherichia coli. Some mutants and stock strains of Escherichia coli K12 were sensitive to acriflavine in the presence of inorganic phosphate but were resistant to acriflavine in its absence. They mutated spontaneously to resistance to acriflavine plus phosphate. The synergistic effect of phosphate on acriflavine sensitivity was increased at high pH values. Genetic analysis suggested that the mutations occurred in the gene acrA. Electron microscopic observation suggested that the presence of acriflavine plus phosphate affected the structure of the plasma membrane and the cytoplasm under it. This structural alteration was not caused by acriflavine alone. Acridine orange plus phosphate can more effectively eliminate the plasmid F8-gal+ than acridine orange alone.


Oxidation of carbon monoxide and methane by Pseudomonas methanica. The oxidation of carbon monoxide and methane by suspensions and ultrasonic extracts of Pseudomonas methanica was studied. A continuous assay for the oxidation of CO to CO2 was devised, using O2 and CO2 electrodes in combination. Stoicheiometries of CO-dependent CO2 formation, O2 consumption and NADH oxidation, and the partial stoicheiometries of methane-dependent NADH oxidation, suggest the involvement of a mono-oxygenase in these oxidations. Evidence is presented suggesting methane and CO oxidation are catalysed by a single enzyme system, distinct, at least in part, from the NADH oxidase present in extracts. Ethanol was able to provide the reductant necessary for CO oxidation by cell suspensions, though the metabolism of ethanol by P. methanica was found unlikely to result in substrate-level formation of NADH; the means whereby alcohol oxidation could supply reductant for the mono-oxygenase are discussed.


Systematic desensitization to reduce dream-induced anxiety. A modified version of systematic desensitization was used to reduce the anxiety and negative interpersonal consequences produced by a recurrent aversive dream resulting from events in the real world. The subject, a 16-year-old incarcerated male, was first taught a standard relaxation technique. The subject's dream was divided into 12 hierarchical imaginal scenes. Following initial relaxation, each scene was sequentially introduced and followed by the therapist's suggestion that the subject was still very relaxed. After three sessions with the therapists and several practice sessions by himself, the subject reported no further anxiety to the dream (which continued to occur) and improved relations with the institutional staff. Six months of follow-up showed no recurrence of the anxiety or the subsequent irritability which the subject had initially reported as leading to negative interpersonal relations with the staff.


Experimental evaluation of the spasmogenicity of dopamine on the basilar artery. Arteriograms of the basilar artery reveal that dopamine given intracisternally to dogs can generate cerebral vasospasm. This finding supports a recent hypothesis of others that dopamine may play a role in the pathogenesis of vasospasm, especially since many substances are known which fail to produce such spasm. Compared to blood or prostaglandin E2, however, the spasm induced by dopamine was delayed in onset, less in incidence, and usually less intense. Possible explanations for such experimental differences are discussed.


Fatty acid and ketone body metabolism in the rat: response to diet and exercise. This study was designed to measure the response of key enzymes of ketone body metabolism in heart, skeletal muscle, and liver to diet and exercise, two conditions known to influence ketone body utilization. A 3 (diet: control, high fat, or high carbohydrate) X 2 (kill condition: rested or exhausted) X 2 (training: trained or untrained) factorial design was used to estimate main experimental effects as well as identify significant interactions of the variables. Physical training (treadmill running) was associated with a doubling of the activity of skeletal muscle 3-oxoacid CoA transferase, a key enzyme in extrahepatic ketone body utilization. The activity of the rate-limiting enzyme of liver ketone body production, hydroxymethylglutaryl CoA synthetase (HMG CoA synthetase), was not greatly influenced by training or exhuastive exercise indicating that the metabolic control of the ketosis of exercise may more likely be a function of the supply of fatty acids to the liver rather than the activity of HMG CoA synthetase. Feeding a high fat diet, on the other hand, significantly increased the activity of liver HMG CoA synthetase, indicating that the ketosis of fat feeding may be of a different nature than that of exercise. The results of this study indicate that physical training is associated with biochemical adaptations in ketone body metabolism as well as fatty acid oxidation, and that trained individuals are metabolically better endowed to benefit from the ketosis of exercise than untrained individuals.


Some mechanisms of reduction of carotenoid levels in chickens infected with Eimeria acervulina or E. tenella. The levels of plasma carotenoids were markedly reduced in broiler cockerels infected with Eimeria acervulina or E. tenella. The mechanisms of this depigmentation differed between the two species, being primarily associated with interference of absorption of xanthophyll (carotenoids) from the intestinal lumen with E. acervulina infection and with leakage through the damaged wall of the cecum with E. tenella infection. Chicks reared on an essentially carotenoid-free diet and inoculated with E. acervulina showed no detectable levels of carotenoids in the blood 48 hours after being changed to a diet containing 30 mg of xanthophyll/kg. Conversely, uninoculated chicks and chicks inoculated with E. tenella showed significant and similar increases in plasma levels of carotenoids. Chicks reared on a diet containing xanthophyll and inoculated with E. tenella showed a more rapid decrease in plasma carotenoids than did uninoculated controls when changed to a xanthophyll-free diet. In chicks fed high xanthophyll diets containing chromic oxide, no indication of malabsorption was seen in chicks infected with E. tenella compared with uninoculated controls, whereas chicks inoculated with E. acervulina showed significantly less xanthophyll absorption. Conversely, a marked increase in the xanthophyll : Cr2O3 ratio was observed in the cecal contents of chicks inoculated with E. tenella compared with uninuoculated controls or those inoculated with E. acervulina. Studies of uninoculated chicks pair-fed with chicks inoculated with E. acervulina or E. tenella indicated that the decrease in plasma carotenoids and increases in intestinal pH are not associated with the reduced intake of feed associated with infection. The studies involving uninoculated birds with reciprocal chagnes between high and low xanthophyll diets indicated that plasma carotenoids are a more rapid and sensitive means of measuring changes in pigmentation levels than are visual skin scores carotenoid levels from the skin.


Metabolic studies on the development of ethanol-induced fatty liver in KK-Ay mice. Mechanisms involved in the development of the alcoholic fatty liver in KK-Ay mice were investigated. Incorporation studies using [14C]acetate and [3H]palmitate indicated that the half-life of hepatic triglycerides was doubled in the ethanol-ingesting mice, and utilization of the exogenous fat was significantly increases as compared with that of the control. No persistent alteration was recognized in hepatic oxidation of palmitate, as estimated by in vitro experiments using liver slices obtained from control and ethanol-drinking mice. Enzymic studies indicated that the activities of acetyl COA carboxylase, ATP citrate lyase, malic enzyme, and 6-phosphogluconate dehydrogenase were increased with ethanol drinking. The increment in hepatic triglycerides accumulated during ethanol ingestion was largely accounted for by palmitoleic, oleic, and linoleic acids. These findings demonstrated an augmentation in hepatic lipogenesis as well as an increased utilization of exogenous fats. Ethanol drinking did not cause any appreciable change in plasma triglyceride level and metabolism of adipose tissue. In summary of the present studies, accelerated lipogenesis and increased utilization of the dietary fats may be possible causal factors in the alcoholic fatty liver of KK-Ay mice.


Renal response to acid loading in the developing lamb fetus, intact in utero. Response of the fetal kidney to metabolic acidosis was studied in five fetal lambs, 115-125 days gestation, in order to evaluate the renal contribution to elimination of hydrogen ion during intra-uterine development. Experiments were conducted on healthy unanesthetized fetuses, intact in utero, with catheters implanted at hysterotomy into a fetal femoral artery and vein and into the bladder via the urachus, four or more days prior to the study. A metabolic acidosis was induced by infusion of isotonic lactic acid, 15 m mole/kg, intravenously over a period of 90 minutes. Serial arterial samples were taken and urine collected in fractions before, during and for three hours following the infusion, for measurements of pH, bicarbonate, lactate and electrolytes as well as urine output. During the infusion, urine pH fell from 6.65 to 6.25 and was 6.34 three hours later (Figs. 1 to 4, Tabs. III to IV). Lactic acid infusion caused a prompt increase in urine output from a mean rate of 0.12 to a maximum of 0.28 ml/kg/min at the end of the infusion, returning to control rates three hours later. Lactate excretion increased from 0.05 to a maximum of 4.6 mumole/kg/min at the end of infusion; titratable acid increased from 0.22 to a maximum of 4 muEq/kg/min; the rates of excretion of lactate and titratable acid were still higher than control at the end of three hours. Ammonia excretion increased from 0.21 to a maximum of 0.56 muEq/kg/min three hours after the end of infusion. The acid infusion caused a small but significant fall in excretion of bicarbonate. During the 90 minutes of infusion and over the following three hours, about 800 mumole lactate was excreted while net acid excretion over the same period was no more than half that amount. The diuresis was also accompanied by a net loss of sodium and chloride, the excretion of these ions increasing more than threefold following acid infusion; excretion of potassium decreased to one-third its rate prior to the infusion. During the 90 minutes of infusion, blood pH fell from 7.36 to 7.13, base deficit rose from 3.8 to 16.4 mEq/L and lactate rose from 2.2 to 14.8 mM/L; there was also a small but significant rise in both blood PCO2 and PO2 (Figs. 1 to 2, Tabs. I to II). During the following three hours of recovery, pH rose gradually to 7.29, base deficit and lactate fell to 7.4 mEq/L and 8.7 mM/L respectively. Since renal excretion of net acid and lactate was small, the decrease in blood base deficit and lactate levels during the recovery must therefore be mainly due to equilibration in various fetal compartments as well as placental transfer. These experiments indicate that, in the lamb fetus, intact in utero, the kidney although limited by immaturity of several mechanisms, is capable of responding to an acid load and thus can make a small contribution to fetal homeostasis. The increase in excretion of net acid is accompanied by loss of sodium and chloride in the urine.


Recognition and significance of maternogenic fetal acidosis during intensive monitoring of labor. FHR monitoring and microanalysis of fetal blood are mutually complementary procedures, and optimal knowledge of the fetal state is achieved by making use of both, the former for the preliminary screening of all cases at risk and the latter for the purpose of deciding on obstetric management where pathological changes are evident in the FHR. The major difficulty in obtaining a precise value for the fetal acid-base balance lies in the occurence of "falsely abnormal" cases, i.e. cases in which the fetal pH falls during labor but the clinical condition at birth is good (APGAR greater than or equal to 7). In our own series the incidence of such cases among fetuses at risk was 11.2% (Tab. I). In the majority of these cases the fetal acidosis is thought to be a result of increased metabolic acidosis in the mother (maternogenic fetal metabolic acidosis). The importance of the maternogenic fetal acidosis during labor lies in the fact that unless it is recognised, rapid extraction of the fetus will appear necessary on clinical grounds, although it is in fact unnecessary, since this form of acidosis has no adverse effect on the fetus. Various parameters have been proposed for the differential diagnosis of the maternogenic fetal acidosis. These include the feto-maternal difference in base deficit (F/M deltaBD), the materno-fetal differences in pHqu 40 (M/F deltapHqu 40) the materno-fetal difference actual pH (M/F actual deltapH), and the materno-fetal difference in base deficit of the extra-cellular fluid (M/F deltaBDHb5). A critical analysis of these parameters has been carried out on the results of microtests performed during a 5 year period (1968-1972) at the First Clinic of Obstetrics and Gynecology of Milan University. The cases comprised 59 regarded as normal (normal course of pregnancy, spontaneous commencement of labor at term, clear amniotic fluid, regular FHR, spontaneous birth, APGAR at 90 sec between 8 and 10, weight at birth greater than 2500 g), and 335 considered to be at risk (maternal disease, presence of meconium stained amniotic fluid and/or abnormal changes in FHR). In all of these cases the FHR was recorded by cardiotokography, and the tracings were interpreted according to HON. Microsamples of blood were taken from both mother and fetus during labor and the following determinations were carried out: actual pH, pHqu 40, Hb concentration, hemoglobin oxygen saturation, base deficit Hb5 (BDHb5). The maternofetal differences were then calculated. The same determinations were carried out on samples of maternal blood and of arterial and venous cord blood taken immediately after delivery. The clinical condition of the infant was evaluated by the APGAR score at 90 seconds after birth.


Solubilization and stabilization of the cytotoxic agent coralyne. Kinetic studies were carried out on the ring opening of the quaternary nitrogen cation, coralynium ion (I), to yield 6'-acetylpapaverine (III), on the cyclization of III to yield I, and on a photochemical reaction undergone by I in aqueous solutions exposed to visible light. From the results, it was concluded that: (a) I and III are in facile equilibrium in aqueous solution but appreciable amounts of III do not exist in dilute solutions with pH values below 10: (b) the photochemical reaction of I in water (presumably a photohydration) can be reversed by lyophilization, by heatiing, and by increasing the pH of solutions to values greater than 12; (c) the photochemical reaction of I can be inhibited by protecting the aqueous solutions from visible light, and the rate in the presence of light can be reduced by increasing the concentration of I in the solution; and (d) although the chloride and sulfoacetate salts of I react identically and have similar solubilities in water, it is possible to prepare more concentrated and, hence, more stable solutions of the sulfoacetate salt by including sodium hydroxide in the solvent. The solubility of coralyne chloride remains about the same in dilute sodium hydroxide as in water.


Solvolysis of a substituted imidazoline, mazindol. Hydrolysis of mazindol to form 2-(2-aminoethyl)-3-(p-chlorophenyl)-3-hydroxyphthalimidine was followed spectro-photometrically in aqueous solutions at temperatures between 37 and 70degree, pH values up to 7.6, and an ionic strength of 0.2. The effects of acetate, formate, and phosphate buffers as well as ionic strength on the observed rate constants were investigated. An interesting nonlinear dependency of the kobs with buffer concentration was noted. The velocity constants declined with increasing hydrogen-ion concentration; the log k-pH profile and rate law are given along with other relevant data.


Complexation in formulation of parenteral solutions: solubilization of the cytotoxic agent hexamethylmelamine by complexation with gentisic acid species. The apparent solubility of hexamethylmelamine in aqueous solutions suitable for intravenous use was increased by complexation with gentisic acid. Studies were carried out in the pH 0-8 range. Unprotonated hexamethylmelamine did not form complexes with the gentisate ion, while the hexamethylmelammonium ion appeared to form several different complexes with both the gentidate ion and gentisic acid. Two different solid complexes were isolated and characterized. The solubility increases observed at pH 3.5-5.0 are described by mathematical relationships involving the stability constants of some postulated complex species. From these results, sultable formulations for use as parenteral solutions are proposed. The increase in the apparent aqueous solubility of hexamethylmelamine in such formulations may range from five- to 90-fold, depending upon the pH and total gentisateion concentrations.


Inhibitory effect of dioctyl sodium sulfosuccinate on trypsin activity. The inhibitory effect of dioctyl sodium sulfosuccinate on the proteolytic activity of trypsin was investigate over the pH 6-8 range. The antitryptic activity was determined using two different substrates: casein and N,alpha-benzoyl-DL-arginine-p-nitroanilide hydrochloride. The mechanistic studies revealed the substrate-inhibitor interaction to be the overall major mechanism of inhibition. This interaction was shown to involve substrate depletion, probably involving some primary sites of the natural substrate casein. Some inhibition was also shown to be due to an interaction between the enzyme and the inhibitior molecules. The interactions of the inhibitor with the enzyme and the substrate were irreversible. The possible therapeutic significance of the inhibitory effect of the surfactant is discussed.


In vitro adsorption of diphenoxylate hydrochloride on activated charcoal and its relationship to pharmacological effects of drug in vivo. I. The adsorption of diphenoxylate hydrochloride, a potent antidiarrheal agent, on activated charcoal powder was studied in vitro. Langmuir adsorption isotherms were established at pH 4 and 7, and the maximum adsorption capacity of charcoal for this drug was estimated using these values. Activated charcoal modified the bioavailability of diphenoxylate hydrochloride in vivo. The antipropulsive action of diphenoxylate in the mouse was strongly inhibited in the presence of activated charcoal. A comparative evaluation of charcoal and chromium oxide used as inert, nonabsorbable markers revealed that chromium oxide may be the marker of choic in GI transit studies in laboratory animals since it does not influence the bioavailability of diphenoxylate hydrochloride.


Binding of bile acids to cholestyramine at gastric pH conditions. The binding of bile salts to cholestyramine was studied under varying conditions of pH and added electrolyte. The taurine-conjugated bile salts were strongly absorbed by the anion-exchange resin at low pH and in the presence of chloride anions. Glycocholic acid binding was very weak at low pH but increased strongly with increasing pH. The presence of chloride ions strongly decreased the amount of glycocholate bound by the anion-exchange resin.


Amodiaquin accumulation by mouse erythrocytes infected with Plasmodium berghei. [14C]amodiaquin accumulation by washed erythrocyte preparations was characterized to permit comparisons with chloroquine accumulation. Erythrocytes infected with Plasmodium berghei CS (chloroquine-susceptible) accumulate amodiaquin by a saturable process that has an apparent dissociation constant for amodiaquin of 7.6 X 10(-8) M and is competitively inhibited by chloroquine, quinine and quinacrine, as is the process of chloroquine accumulation. Within experimental error, the K1 of 8 X 10(-7) M estimated for chloroquine is the same regardless of whether the drug being accumulated is [14C]amodiaquin or [14C]chloroquine. Likewise, the K1 for amodiaquin is the same regardless of which drug is being accumulated. In addition, glucose stimulates and hydrogen ion, cold or interruption of glycolysis inhibits amodiaquin as well as chloroquine accumulation. These findings are evidence that a single process serves to accumulate both drugs. In the absence of substrate, erythrocytes infected with P. berghei CR (chloroquine-resistant) accumulate twice as much amodiaquin as chloroquine, and they accumulate more amodiaquin than do erythrocytes infected with P. berghei CS. These differences occur because P. berghei CR infects polychromatophilic erythrocytes possessing a high-affinity, substrate-independent process of accumulation to which amodiaquin has greater access than chloroquine. In the presence of glucose, amodiaquin accumulation by erythrocytes infected with P. berghei CR, when plotted as a function of amodiaquin concentration in the medium, describes a sigmoid curve.


The neurochemistry of Parkinson's disease: effect of L-dopa therapy. Post-mortem brain material from control and Parkinson's disease patients was examined to elucidate further the neurochemistry of this disease and to determine the mechanism of action of L-dopa as a therapeutic agent. The activities of L-aromatic amino acid decarboxylase (dopa D), tyrosine hydroxylase, monoamine oxidase and catechol-O-methyl transferase were examined; in addition the tissue levels of dopa, 3-O-methyldopa, dopamine (DA) and homovanillic acid (HVA) were determined. In the non-dopa-treated Parkinsonian patients, the greatest decreases were detected for striatal DA and dopa D, with homovanillic acid and tyrosine hydroxylase levels showing a lesser change. The activities of monoamine oxidase and catechol-O-methyl transferase in the striatal nuclei were not different from the controls. The putamen was consistently the most severely affected region. Dopa and 3-O-methyldopa were detectable in all brain areas only in those patients treated with L-dopa shortly before death. The mean concentrations of DA in the striatum of these patients were 1) 9 to 15 times higher than those in non-dopa-treated patients, 2) related to the time before death of the last dose of L-dopa and 3) greater in the striatum of patients clinically classified as "good responders" as compared to "poor responders." Although L-dopa therapy increased homovanillic acid levels in all brain areas, a preferential increase was observed in the striatum. It was concluded that L-dopa's principal therapeutic effects in Parkinson's disease are consistent with its transformation to DA in the striatum.


Studies on the mechanism of depletion of striatal dopamine by alpha-methyl-m-tyrosine. These experiments were designed to study the mechanism of depletion of dopamine (DA) in the striatum produced by alpha-methyl-m-tyrosine (alpha-MMT). alpha-Methyl-m-tyramine (alpha-MMTA), the metabolite of alpha-MMT, appears to be the active DA-depleting agent, since the administration of a decarboxylase inhibitor before alpha-MMT markedly reduced both the formation of alpha-MMTA and the depletion of DA. After injection of alpha-MMT (100 mg/kg i.p.), the striatal concentration of homovanillic acid (HVA) rose by 41% at 1 hour. This is probably due to an increase in DA metabolism, since alpha-MMT markedly enhanced the decline of DA produced by alpha-methyl-p-tyrosine (alpha-MPT). At 2, 3 and 4 hours after alpha-MMT, the concentration of HVA and dihydroxyphenylacetic acid was below control level. The decrease in dihydroxyphenylacetic acid is due partially to a decreased formation of dihydroxyphenylacetic acid from DA. In striatal slices, both alpha-MMT and alpha-MMTA decreased the formation of 3H-H2O and the accumulation of 3H-DA from 1-3,5-3H-tyrosine. Alpha-MMT did not alter the specific activity of 3H-tyrosine or release 3H-DA from the slices, but it did inhibit the activity of tyrosine hydroxylase in striatal homogenates at low concentrations of tyrosine (10 muM). Alpha-MMTA released both newly synthesized and exogenously accumulated 3H-DA from striatal slices. At low concentrations of alpha-MMTA, the percent reduction in 3H-H2O was much greater than the percentage of 3H-DA released into the medium. However, at higher concentrations, the inhibition of 3H-H2O reached a maximum while 3H-DA release kept increasing. These results suggest that both inhibition of tyrosine hydroxylase activity and DA release from storage sites by alpha-MMTA may account for the depletion of DA produced by the injection of alpha-MMT.


Characteristics of gastric inhibition by acidification of oxyntic gland area. 1. Gastric acid responses to the test meals were measured in the Heidenhain pouch, gastric and pancreatic fistula dogs, using the intragastric titration method, and monitoring the rate at which a solution of 1-0 N-NaOH had to be added to maintain the pH of the gastric content constant at pre-selected values ranging from 5-0 to 1-0. In this way the pH profile of the gastric acid and pepsin responses to a liver extract meal kept in the Heidenhain pouch or gastric fistula as well as to exogenous stimuli such as histamine, pentagastrin or Urecholine could be determined. 2. A liver extract meal adjusted to pH 5-0 produced a potent and pressure-related stimulation of acid secretion from the Heidenhain pouch without any change in secretion from the main stomach and pancreas or in the serum concentration of immuno-assayable gastrin. 3. Graded decrease of the liver extract meal pH to below 5-0 resulted in the pH-dependent inhibition of gastric acid output, which at pH 1-0 was only about 30% of the value attained at pH 5-0. Acid secretion from the Heidenhain pouch induced by exogenous stimuli such as histamine, pentagastrin or Urecholine also showed gradual decrease when the pH of the pouch content was decreased in sequential order from 5-0 to 1-0. This pH-dependent inhibition was accompanied by an increase in pepsin secretion. 4. The pH-dependent inhibition of the Heidenhain pouch response to the liver extract meal was not altered by topical application of a local anaesthetic and atropine or by the intravenous infusion of large doses of atropine, secretin or metiamide, which were shown to cause a marked inhibition of the main stomach response to the liver meal. 5. The results indicate that there is a local and gastrin-independent inhibition mechanism of gastric acid secretion activated by an acidified meal making contact with the oxyntic gland area.


The influence of pH on equilibrium effects of tetrodotoxin on myelinated nerve fibres of Rana esculenta. 1. The experiments were done on single nodes of Ranvier of Rana esculenta. The effects of tetrodotoxin and H ions were determined either by the reduction of the maximum rate of rise, VA, of action potentials evoked with threshold stimuli or in the voltage clamp by the decrease of the peak Na permeability, PNa. 2. With the tetrodotoxin sample used throughout the investigation the equilibrium dissociation constant, KT, of the toxin-receptor reaction at neutral pH was determined to be 2-8 nM. Between 1-55 and 15-5 nM tetrodotoxin the normalized value, A, of VA, was found to be related to the normalized toxin concentration cT = [TTX]/2-8 nM by the empirical equation log [(1-A)/A] = 1-22 log cT-0-573. 3. On increasing the pH (up to 8-8) the effect of tetrodotoxin diminished as revealed by an increase in A. The apparent reduction of cT (as calculated from A) suggests that the toxin is active only in its cationic forms. 4. Weakly acid tetrodotoxin solutions (7-3 less than pH less than or equal to 5-5) reduced A to a lesser degree than did neutral toxin solutions in spite of the inherent depressing effect of acid pH on A (A = 0-5 at about pH 5-5). In more acid toxin solutions A decreased again and at pH 4-6 it was about equal to the value in toxin-free solution. 5. When, after equilibrium in an acid toxin solution, the perfusate was suddenly changed to neutral Ringer solution A jumped to a higher value A' as measured 1 sec after the switch. Since the blocking effect of hydrogen ions subsided within a fraction of a second while the time constant of the toxin washout is of the order of 1 min, A' reflects the number of Na channels blocked by tetrodotoxin at acid pH. 6. In acid toxin-free solution the peak PNa as obtained in voltage clamp experiments was reduced by a voltage-dependent factor (cH + 1)-1 with CH = [H+]/KH(E) and KH(E) = 2-04 muM exp (0-34 EF/RT). Adding tetrodotoxin resulted in another reduction by a constant factor p'T. 7. Experiments employing various combinations of toxin concentration (3-1-93 nM) and pH values (7-3-5-2) confirm the decreased toxin effect at low pH. Moreover, p'T was smaller (the additional toxin effect larger) when the membrane had been kept depolarized and thus cH reduced during equilibration. This suggests that tetrodotoxin cations and H ions compete for the same blocking site. A quantitative fit, however, requires additional assumptions.


The influence of pH on the rate of tetrodotoxin action on myelinated nerve fibres. 1. The experiments were done on single myelinated nerve fibres of Rana esculenta. The rates of toxin effect were studied either by measuring the maximum rate of rise, VA, of repetitively evoked action potentials or by measuring Na currents during periodic impulses in the voltage clamp. 2. VA measurements showed that in alkaline solutions (pH up to 8-8) the offset rate was unchanged while the onset was slowed in quantitative agreement with an assumed decrease in the active cationic form of tetrodotoxin. 3. Both VA measurements and those in the voltage clamp revealed a decrease in T'off, the offset time constant and in increase in the onset time constant, T'on, as the pH was lowered. 4. For tetrodotoxin concentrations, [TTX], up to 400 nM and pH values down to 5-3 the simple relation T'on/T'off = p'R held, where p'T is the constant factor by which the Na permeability was reduced at equilibrium with a given [TTX]. 5. The agreement between kinetic and equilibrium results was also valid when, at constant [TTX] and pH. p'T was modified by the holding potential during equilibration. 6. No unequivocal explanation of the results can be given but some of their features resemble acid catalysis.


The formation of synapses in amphibian striated muscle during development. 1. A study has been made of the formation of synapses in developing reinnervated and cross-reinnervated amphibian twitch muscles which receive either a focal (iliofibularis) or a distributed (sartorius) innervation from 'en plaque' nerve terminals using histological, ultrastructural and electrophysiological techniques. 2. During the development of the tadpole through metamorphosis to the adult frog, the sartorius myofibres increased in length at about twice the rate of the iliofibularis myofibres, due to a fast rate of growth at their insertions on to the pelvic tendon. 3. The short iliofibularis and sartorius myofibres of young tadpoles (800 mum long) possessed only a single synapse and the iliofibularis myofibres did not receive any further innervation during development. However the sartorius myofibres received further transient innervation on the new muscle laid down during development at the fast growing pelvic insertion, until the distance between the original synapse formed on the myofibres and the synapse at the pelvic end of the muscle was about 12 mm. 4. During development synapses possessed either skewed, multimodal, or unimodal m.e.p.p. amplitude-frequency distributions; the intervals between m.e.p.p.s. were not distributed randomly according to a Poisson process, as m.e.p.p.s. of similar amplitudes tended to be separated by very short intervals; the unit-size e.p.p. had a similar amplitude-frequency distribution as the m.e.p.p.s. if these had a unimodal distribution. 5. Reinnervation or cross-reinnervation of the sartorius and the iliofibularis muscles in adults or at a late stage of development simply reconstituted the normal focal and distributed innervation patterns of the muscles, as found in the control muscles of the contralateral and unoperated legs. 6. These observations on synapse formation in amphibia are consistent with the hypothesis that during development the axon making the initial synaptic contact on the muscle cells induces a property over a length of muscle membrane adjacent to this site which makes it refractory to synapse formation; thus during reinnervation or cross-reinnervation of adult muscles this refractory property constrains synapse formation to these sites.


Comparative studies of Trypanosoma vespertilionis Battaglia and Trypanosoma dionisii Bettencourt & França. In diphasic blood agar media Trypanosoma vespertilionis developed spheroid clusters as compared to rather long, sausage-shaped (sometimes branched) clusters formed by Trypanosoma dionisii. The former species attained a greater population density (approximately 6 X 10(7) organisms/ml) than the latter (approximately 2 X 10(7) organisms/ml). Greater numbers of epimastigotes, some in active binary divisions, were observed during the logarithmic phase of growth, and morphologic changes occurred during cultivation which correlated with increased acidity and a depletion of glucose. Maximum numbers of trypomastigote forms were found during the stationary and early death phases. Most of the forms observed after 20 days were sphaeromastigotes. Glucose concentrations decreased to 0 M in T. vespertilionis and to 4.4 X 10(-5) M in T. dionisii cultures during the stationary and death phases. By the 12th day of incubation cultures of T. vespertilionis were more acid (pH 5.5) than those of T. dionisii vespertilionis and T. dionisii contained common and specific antigens. At least 2-3 common antigens were detected in extracts reacted against heterologous antisera. Specific antigens were observed as nonidentical lines formed by extracts reacted against homologous and heterologous antisera and with antisera absorbed with heterologous antigens. At least 2 specific antigens were evident in extracts of T. vespertilionis and 1 in extracts of T. dionisii.


The use of psychotropic drugs in general practice. A report of a year's survey. All the psychotropic tablets prescribed by a general practitioner in one year are itemised. The patients who received them are separated into diagnostic groups and their treatment is analysed by the number of attendances, prescriptions, and duration of therapy. Hospital referrals and overdoses over a longer period are recorded. My policy in psychiatric conditions is indicated.


N-Isopropyl derivatives of dopamine and 5,6-dihydroxy-2-aminotetralin. Secondary and tertiary amino homologs of the title compounds have been prepared, bearing an N-isopropyl group. In peripheral evaluation, certain members of the series exhibited beta-adrenergic agonist effects of lower activity than isoproterenol. N-Methyl-N-isopropyl-5,6-dihydroxytetralin exhibited marked properties consistent with its being an alpha agonist, and it is concluded that introduction of considerable bulk about the nitrogen of a catecholamine does not a priori destroy alpha-agonist effects. The compounds qualitatively paralleled the effects of dopamine in assays based upon direct intrastriatal administration in rats, although they were less potent than dopamine.


Phospholipases. III. Effects of ionic surfactants on the phospholipase-catalyzed hydrolysis of unsonicated egg lecithin liposomes. Apparent values of Km and Vmax have been measured for catalysis of hydrolysis of unsonicated egg lecithin liposomes, activated through addition of 0.4 M n-hexanol, by phospholipases A2 from bee and snake venoms and by phospholipase C from Clostridium welchii as a function of the concentration of three surfactants: hexadecylamine, hexadecyltrimethylammonium bromide, and dihexadecyl phosphate. For all three enzymes, values of Km and Vmax show little or no dependence on the concentration of these ionic surfactants, demonstrating that the liposomal surface charge is not a crucial factor in determining susceptibility to phospholipase-catalyzed hydrolysis.


Negative potential level in the outer layer of the toad skin. The isolated skin of the toad Bufo marinus ictericus when impaled from the outer surface by glass microelectrodes filled with 3 M KCl shows a voltage profile which is a continuous function of the depth of impalement. The superficial intraepithelial potential difference measured with reference to the external solution (PDi) is negative with NaCl-Ringer's solution on both sides of the skin, displaying a minimum of -26.7+/-3.6 mV at 6+/-2 mum. Null value is obtained at 19+/-3 mum, with positive values for deeper impalements. Indications of cell impalements (abrupt voltage and resistance jumps) were frequently observed at sites deeper than 25 mum from the outer surface. Measurements of the electrical resistance between the microelectrode and the external solution, made with single- and double-barreled microelectrodes, showed great discrepancies, which may be attributed to distinct pathways of different resistances in the stratum corneum. PDi measured at a depth of 5 mum was a logarithmic function of Na2SO4 or K2SO4 concentration in the external solution, increasing in negativity with a reduction in concentration. Substitution of Na by K in the external solution had only minor effects on PDi. Acidification of the external solution from pH 9 is accompanied by a reduction in the negative value of PDi. At pH 3 PDi was positive. PDi was interpreted as a diffusion potential at the tip of the microelectrode due to KCl diffusion from the electrode into the matrix of the stratum corneum. Differences in K and Cl mobilities, responsible for the origin of PDi, were attributed to fixed charges in the matrix of the stratum corneum, with density and polarity determined by their degree of proponation, controlled by the hydrogen ion concentration of the external solution. Skin potential, short-circuit current and their relationship to PDI were discussed.


The avian erythrocyte: a study of fixation for electron microscopy. The quality of ultrastructural preservation of the avian erythrocyte achieved using various fixation techniques is evaluated. Different combinations of initial fixatives, buffers and post-fixation procedures were tested as well as variations in fixative osmolarity, pH and temperature. Of the commonly used initial fixatives (glutaraldehyde, acrolein and formaldehyde), 2% glutaraldehyde, alone in a slightly hypertonic buffer containing divalent ions, produced optimum erythrocyte preservation. The osmolarity was balanced using a non-electrolyte such as a sucrose. The addition of 12% hexylene glycol to the buffer solutions also improves erythrocyte preservation, as evidenced by the increased stability of the marginal microtubules, microfilaments and proteinaceous material. The use of Spurr low-viscosity epoxy resin enables the cells to be collected using low gravitational centrifugation.


Hemoglobin solution and the oxyhemoglobin dissociation curve. 1) A study was carried out to determine the oxyhemoglobin dissociation curve of stroma-free hemoglobin solution and factors which influence it, (pH; 2,3 DPG). 2) To simulate acute volume replacement, dilution experiments, in vitro, were performed employing both hemoglobin solution and Ringer's lactate in whole blood. 3) It was determined that stroma-free hemoglobin solution has a left-shifted oxyhemoglobin dissociation curve which responds to pH change but not to the addition of 2,3 DPG. 4) The dilutional effect of hemoglobin solution when mixed with whole blood in volumes up to 50% was to left-shift the oxyhemoglobin curve, unlike the effect of Ringer's lactate (no change). 5) This may have importance in the hemodynamic compensatory response to acute normovolemic anemia.


Effect of antihistamine-antiserotonin and ganglionic blocking agents upon increased capillary permeability following burn trauma. Tiny (0.2% TBS), partial thickness, non-contact radiant heat burns in guinea pigs resulted, within 3 hours, in significant edema formation and protein leakage at the site of the injury. Areas of skin distant to the burn also showed an increase in water content but no protein leakage. Pretreatment of the animals with either chlorisondamine hydrochloride or a mixture of methysergide and chlorpheniramine significantly decreased postburn edema formation and protein leakage. Liquid emulsion autoradiography revealed that leakage of protein occurs primarily in the areas of skin adjacent to the panniculus carnosus. The studies suggest that: the increase in vascular permeability that occurs as a consequence of burn injuries is humorally mediated; albumin leakage is limited to the injured tissues; and histamine, serotonin, and presumably catecholamines play significant roles in the development of this phenomenon.


Bacteriophage T4 baseplate components. II. Binding and location of bacteriophage-induced dihydrofolate reductase. The location of T4D phage-induced dihydrofolate reductase (dfr) has been determined in intact and incomplete phage particles. It has been found that phage mutants inducing a temperature-sensitive dfr (dfrts) procude heat-labile phage particles. The structural dfr produced by these ts mutants was shown to assume different configurations depending on the temperature at which the phage is assembled. Morphogenesis of incomplete phage particles lacking the gene 11 protein on their baseplates was found to be inhibited by reagents binding to dfr, such as antibodies to dfr. Further, cofactor molecules for dfr, such as reduced nicotinamide adenine dinucleotide phosphate and reduced nicotinamide adenine dinucleotide, also inhibited the step in morphogenesis involving the addition of gene 11 product. On the other hand, inhibitors of dfr, such as adenosine dephosphoribose, stimulated the addition of the gene 11 protein. It has been concluded that the phage-induced dfr is a baseplate component which is partially covered by the gene 11 protein. The properties of phage particles produced after infection of the nonpermissive host with the one known T4D mutant containing a nonsense mutation in its dfr gene suggested that these progeny particles contained a partial polypeptide, which was large enough to serve as a structural element.


Uukuniemi virus contains an RNA polymerase. An RNA-dependent RNA polymerase activity has been found associated with Uukuniemi virions. The enzyme activity is expressed only after disrupting the virions with the nonionic detergent Triton X-100 and is absolutely dependent on Mn2+, whereas Mg2+ is not required, a finding that distinguishes this polymerase from those of other enveloped minus-strand RNA viruses. Within the range pH 7.2 to 8.5 no distinct optimum was found. The optimum temperature was between 37 and 40 C. The reaction was not inhibited by actinomycin D, rifampin, or DNase, whereas RNase was completely inhibitory. The partially RNase-resistant product consisted of rather small-sized RNA, which contained sequences complementary to Uukuniemi virus RNA as shown by hybridization to the template L, M, and S RNA species of Uukuniemi virus.


Radiographic examination of mandibular lesions in barren-ground caribou. Dental anomalies were observed in 43 of 1,226 barren-ground caribou (Rangifer tarandus groenlandicus) taken between 1966 and 1968. In five of these 43 animals, the mandibles had deformities which radiography showed to be the result of dental abscesses in four cases and probably of a trauma in the other. The absence of actinomycotic lesions of the jaw bones of these 1,226 animals, and of more than 500 examined previously, indicates that "lumpy jaw" is rare in barren-ground caribou. The authors suggest the use of radiography to determine the nature of bone growth on skeletal remains, in the absence of soft tissues for examination for Actinomyces, either microscopically or by cultural methods.


Experimental diarrhea in cynomolgus monkeys by oral administration with Clostridium perfringens type A viable cells or enterotoxin. Purified C. perfringens type A enterotoxin fed orally in an amount of 5 mg caused both vomiting and diarrhea in the monkey only when the gastric juice had been neutralized. Exposure of enterotoxin to pH 4.0 or below rapidly destroyed the activity. All three monkeys receiving sodium bicarbonate and 2.4 X 10(10) viable cells grown in DS medium developed diarrhea, and only one of them vomited once. The diarrhea lasted for 13, 18 and 19 hr. The symptoms were similar to those reported in human cases of C. perfringens food poisoning. These results have verified the general notion that C. perfringens food poisoning should be categorized as a true "intravital intoxication". The reversed passive hemagglutination test detected enterotoxin directly in most fecal samples. This method may be applicable for diagnosis of human cases of C. perfringens food poisoning. Neither enterotoxin nor anti-enterotoxin was detected in serum samples taken from any monkey up to 21 days after the challenge. We are tempted to conclude, therefore, that no significant amount of C. perfringens enterotoxin is absorbed from the intestine.


[Effect of peripheral counterpulsation on the body of an animal with intact heart]. The effect of lasting peripheral counterpulsation upon the haemodynamics and the main biochemical factors of the blood was studied in 14 dogs with intact hearts. In cases of significant tachycardia counterpulsation in a 1:2 regimen results in only a partial reduction of the resistance to the cardiac output. This does not always permit to prevent the formation of the phenomenon of an elevated myocardial contractility. The haemodynamic conditions are most optimal in a 1:1 regimen of counterpulsation. Slowing down of the cardiac rhythm was achieved by means of hypothermia.


[Regulation of interrelation between pulmonary ventilation and circulation]. In acute experiments on 92 cats anesthetized with Urethane and kept under controlled respiration the mechanism of tonic activity of the pulmonary vessels was studied in the presence of a decreased partial pressure of oxygen in the alveoli. The tonicity of the pulmonary vessels was recorded during autoperfusion of the vessels of the posterior lobe of the left lung by means of a perfusion pump. Simultaneously, the pressure in the common carotid artery was recorded and the oxygen saturation of the blood was measured. Pharmacological analysis was used for the study of the mechanism of the pressor reaction of the pulmonary vessels under hypoxic hypoxy, and it demonstrated that the pression of the pulmonary vessels that develops under alveolar hypoxy is less distinct under the effect of the ganglioblocking agent Benzo-hexonium, as well as the myotropic agents Papaverine and Chloracizine. The above reaction was significantly inhibited by the blocking agents of the D- and M-serotonin-reactive structures -- Dihydroergothamine, Morphine and Novocain, and it was completely lacking against the background of the action of Izadrine and Dimedrol -- blocking agents of serotonin- and histamine-reactive structures. It can be supposed that the D- and M-serotonin-reactive, and probably also the histamine-reactive structures participate in the regulation of the interrelationship of ventilation and pulmonary circulation.


Multiple cyclic nucleotide phosphodiesterases in rat kidney. Using DEAE-cellulose chromatography and Agarose gel filtration we have partially purified a low Km cyclic adenosine monophosphate (AMP) phosphodiesterase from the 100,000 X g supernatant of rat kidneys. The characteristics of this enzyme included a Km of approximately 4 muM a pH optimum of around 8.0 and a requirement for magnesium. This preparation should be suitable for investigation of possible effects of hormones, drugs and cellular constituents on the cyclic AMP pathway through any direct effects on the low Km enzyme. We have also demonstrated a nonspecific, high Km cyclic nucleotide phosphodiesterase and possibly a specific cyclic guanosine monophosphate (GMP) phosphodiesterase in the soluble fraction from rat kidneys.


Effect on aging on plasma renin and aldosterone in normal man. The influence of aging on the renin-angiotensin-aldosterone system was evaluated by comparing young (20 to 30 yr) with elderly (62 to 70 yr) healthy subjects. Despite comparable body sodium-fluid balance in the two age groups, serum renin concentration, plasma renin activity and aldosterone concentrations were lower in the elderly. The age-related decreases in circulating renin and aldosterone concentrations were slight while subjects were supine and receiving normal sodium intake; when upright and during sodium depletion, they were more pronounced. Inverse renin-blood pressure interrelations were noted during two of four study conditions involving normal sodium intake or mild sodium depletion (r = --0.44 and --0.47, respectively), but not during progressive sodium depletion. Plasma renin levels were decreased in the elderly regardless of the presence or absence of an inverse relationship with blood pressure. Aldosterone and cortisol responses to corticotropin infusion were unaltered in the elderly. It is concluded that aging may cause a decrease in circulating renin, with parallel lowering of plasma aldosterone concentrations.


[Immune mechanisms in uremia].

[Immune mechanisms in uremia]. There is both clinical and experimental evidence that cellular and humoral immunity are suppressed in patients with renal insufficiency: observations in organ transplantation and in vitro stimulation of lymphocytes from uraemic patients, investigations of acute and late hypersensitivity reactions, the immune response after active immunization as well as changes of immunoglobulins and lymphatic organs in uraemia are discussed in the paper. The underlying mechanisms are complex and not yet fully understood. Lymphopenia, atrophy of the thymus gland, toxic serum factors, induction of enhancing mechanisms by certain serum fractions and metabolic defects of lymphocytes--all were shown to be involved or at least considered to be. At present, however, it is impossible to define their rank of importance and the exact place they may occupy in the genesis of this type of "natural immunosuppression".


Improvement of renin determination in human plasma using a commonly available renin standard in a radioimmunological method. A new method for the measurement of renin in human plasma is described. The method is based on the introduction of the internationally available renin standard of the Medical Research Council (MRC) London, as a calibration system. Thus, some principal disadvantages of methods expressing results in renin reaction velocity (angiotensin generation rate) only are avoided. Both renins, unknown and standard, react with a sheep substrate preparation and are handled identically throughout the whole procedure including the angiotensin I radioimmunoassay (RIA). The plasma renin concentration (PRC) is given in 10(-6) MRC-renin units (muM/ml). the renin standard is free of angiotensin, angiotensinases, and angiotensinogen; it is stable on storage. Identical enzyme kinetics are shown for both renins. An interference between endogenous and exogenous substrate could be avoided. The potentially harmful influences of proteins from the enzyme incubation mixture of the RIA dose response curve are shown. The use of an angiotensin I calibration system could be omitted. Using a standard renin dilution from 250-0.9 muU/ml also the full biological range is covered. When giving an unrestricted diet the preliminary normal values of PRC are 21.9 +/- 12.6 muU/ml in recumbent and 40.1 +/- 19.8 muU/ml in upright position (n = 16,x +/- s, age 20-35 years). Earlier findings of age-dependency of PRC were confirmed.


[Validity of pH measurements by means of micro pH combination electrodes in blood and other biological fluids (author's transl)]. pH measurements in blood or in media containing either proteins or polypeptides and performed by means of micro pH combination electrodes type N 58 (Schott & Gen., Mainz) yield in a systematic error according to the regression line y = 1.135 chi - 0.842, in the range between pH 5.3 and 8.3. This deviation from the real pH value is independent of the protein concentration and amounts to 0.1-0.2 pH units in the  physiological range. The error does not occur if the pH measurements are performed in media which are free of proteins and polypeptides, respectively. If the electrolyte solution within the reference electrode is replaced (NaCl solution instead of KCl solution) the error is distinctly reduced. For this reason, this deviation should be caused by the variation of the diffusion potential across the platinum junctions.


The effects of drinking on offspring: an historical survey of the American and British literature. Current research on the effects on offspring of drinking during pregnancy has revived interest in an extremely old topic. Observations made during England's Gin Epidemic (1720-1750) were followed by warnings of 19th-century medical writers that parental drinking could damage the fetus. Many concurring studies were reported in the medical literature from 1865 to 1920. Research interest declined during Prohibition, and authorities later discounted the previous work. Recently a relationship between maternal drinking and abnormal morphogenesis has been again described.


[Glutathione (author's transl)].

[Glutathione (author's transl)]. Glutathione plays an important role in biology and medicine. Most cells of plants and animals contain high concentrations of reduced glutathione and a much smaller amount of oxidised glutathione. GSH is important for several metabolic functions of live cells, e.g. the protection of oxidative stress by peroxides, mediation of enzyme reactions, regulation of metabolic events, transport of amino acids across cell membranes via the gamma-glutamyl cycle, elimination of foreign compounds by GSH-conjugation, release of neurotransmitter substances. Irreversible perturbations of the glutathione metabolism may be the reason for severe clinical symptoms of hemolytic anemia or, perhaps, of central nervous disease.


Hyperexcitability in the neural substrate of emotional behavior in cats after alcohol withdrawal. Evidence of a rapid development of alcohol dependence. Substantial and prolonged withdrawal hyperexcitability in the neural substrate for affective defense was revealed by behavioral and electrophysiological measures in cats exposed to moderate to heavy doses of alcohol for periods ranging from 6 to 72 hours. The data are interpreted as indicating a rapid development of physical dependence on alcohol in this portion of the central nervous system.


Interactions of age, sex and long-term alcohol intake in selectively bred strains of rats. The alcohol consumption by five genotypes of rats was studied in two experiments. Alcohol intake was age-dependent in rats bred for high emotional reactivity and avoidance conditionability. Differences in consumption by sex appeared to be primarily due to differences in body weight.


Regional study of acid hydrolases and lysosomal membrane properties in the normal human brain at various ages. Acid hydrolases and lysosomal membrane properties were studied at various ages in the normal human brain. In CSF and four brain regions, the inferior olive, the cerebellar cortex, the caudate nucleus and the frontal cortex were thus beta-galactosidase, beta-glucosidase, alpha-mannosidase, hexosaminidase and acid phosphatase biochemically quantitated at ages varying between 2 and 89 years of age. Also the membrane latency for acid phosphatase was studied in these regions. No major regional quantitative differences were found with regard to the enzymes studied. Their kinetic properties were also defined. There appeared to exist a regional and intra-areal variation in lysosomal membrane permeability. There was, however, no age related increase in total enzyme contents. The possibility significance of these findings are discussed with reference to the aging process.


The late effects of selected immunosuppressants on immunocompetence, disease incidence, and mean life-span. II. Cell-mediated immune activity. The late effects of various immunosuppressive insults on cell-mediated immunity in mice were studied in an attempt to assess the role of immune surveillance in the aging process. Results were obtained using susceptibility to allogeneic tumor cell challenge, graft-versus-host reaction (GVHR), blastogenic response to PHA, a thymus derived T cell-specific plant mitogen, and cytolytic activity against allogeneic tumor cells as measures of immunologic activity. In vivo studies late in life show that resistance to allogeneic tumor cells is significantly decreased in thymectomized mice, whereas those treated with cortisone, cyclophosphamide and sublethal X-ray remain unchanged. Spleen cells from only the thymectomized and the sublethally irradiated mice show reduced activity in the GVHR. No difference is seen in the activity of bone marrow cells. Results consistent with these findings were obtained in in vitro studies. Thus spleen cells from thymectomized or sublethally irradiated mice show decreased activity is response to PHA, whereas no change is seen in spleen cells from other treated groups. Hence, surgical and physical insults are more likely to induce long-lasting immunosuppression in those immunocompetent tissues whose activity normally diminishes with advancing age. Furthermore, the degree of immunosuppression seen in this study is not of the order of magnitude that one could reasonably predict a significant decrease in mean life-span.


Lipoxygenase isozymes of peanut.

Lipoxygenase isozymes of peanut. Lipoxygenase was isolated and partially purified from peanut seed by ammonium sulfate precipitation, gel filtration, and ion exchange column chromatography. Three isozymes of lipoxygenase were identified. Two had pH optima of 6.2, and the other an optimum of 8.3. Molecular weight of each isozyme was 7.3 x 10(4), as determined by gel filtration. The alkaline optimum isozyme was not inhibited by NaCN and was inhibited by CaCl2 except at very low concentrations. The acid optimum isozymes were inhibited by NaCN and were stimulated by CaCl2 concentrations up to ca. 0.7 mM.


Crassin acetate, the principal antineoplastic agent in four gorgonians of the Pseudoplexaura genus. Crassin acetate, a lactonic cembrane diterpene, has been shown to be the principal antineoplastic agent present in the marine invertebrates (gorgonians) Pseudoplexaura porosa, P. flagellosa, P. wagenaari and P. crucis.


[Computer experience and further developments in the respiratory function laboratory (author's transl)]. Reported is on satisfactory results obtained with a small-size computer consisting of punching and scanning device, as well as plain writing machine in the respiratory function laboratory. Developed in on- as well as off-line processing by an own technical staff, a diagnostic and teaching program was established for all respiratory function routine methods with the advantages of a large number of cases examined, elimination of sources of error, considerable supply of data and information, automatic documentation and filing, plain writing, interpretation and evaluation of findings. In continuation of such works also the blood gas analysis has been included. These values as the total of disturbances of the pathophysiological acid-base status are considered and interpreted. Clinical correction is forced in this man-machine dialogue by automatic stops of the whole machinery before going on. Subsequently and in addition are computer alveolar-arterial oxygen pressure gradient, venous shunt and oxygen saturation and expressed utilizing the capacity of the small-size computer. Further developments in the respiratory function diagnostic- and teaching program for small-size computers--not too expensive in the building block principle - are intended.


Phosphate transport in rat liver mitochondria. Kinetics, inhibitor sensitivity, energy requirements, and labeled components. Experiments were carried out to define the kinetic parameters of the major phosphate transport processes of rat liver mitochondria, and to obtain information about the molecular properties of these systems.


L-tyrosine: 2-oxoglutarate aminotransferase induction by hydrocortisone in the thymus of the white rat. Hydrocortisone hemisuccinate within 4 hours after in vivo administration produced an increase in precursor incoporation into rat thymus RNA and proteins in the whole animal. From these results, together with information obtained from measurements of the tyrosine aminotransferase activity and the action of mitomycin C administered one hour before the injection of hydrocortisone, it can be concluded that the increase in tissue level of the enzyme, consequent to hydrocortisone treatment, results from an increased rate of biosynthesis of the enzyme, which participates in the catabolic processes of proteins in glucocorticoid sensitive thymus cells.


Identification of the 30 S protein adjacent to peptidyl transferase catalytic center of Escherichia coli ribosomes. Iodoacetylphenylalanyl-tRNAPhe was used as an affinity label to localize the ribosomal components involved in the peptidyl transferase catalytic center of Escherichia coli ribosomes. When labeling was carried out at pH 5.0, the affinity label could specifically label the ribosomal components which comprise the catalytic center. Analysis of ribosomal proteins which had reacted with the affinity label revealed that a 30 S subunit protein, S 20, was located at or near to the ribosomal binding site of the 3'-terminus of aminoacyl- or peptidyl-tRNA.


The value of a histamine H2-receptor antagonist in the management of patients with the Zollinger-Ellison syndrome. Inhibition of acid secretion by an H2-receptor antagonist (metiamide) was assessed in three patients with the Zollinger-Ellison syndrome. Metiamide (200 or 300 mg) inhibited acid secretion transiently (2 1/2 hours) by 85 to 100 per cent in all patients. Although anticholinergic drugs alone inhibited acid secretion by only 0 to 35 per cent in these patients, the combination of metiamide and anticholinergic markedly prolonged the inhibitory effect of metiamide. Total gastrectomy was refused by one patient, and was impossible in another; both were treated with metiamide and anticholinergic for five and 10 months. A third patient was treated with metiamide and anticholinergic for three weeks in preparation for total gastrectomy. Ulcer pain and diarrhea disappeared, and each gained weight. H2-receptor antagonists may be useful in the treatment of some patients with the Zollinger-Ellison syndrome.


Histamine receptors in the vasculature of the rabbit ear. Histamine has a dual action on the isolated perfused ear preparation of the rabbit. The amine induced a dose-dependent rise in perfusion pressure when the preparation was perfused with Krebs' solution. This pressor response was reversed to a depressor effect when meypramine was added to the perfusion fluid. This depressor effect of the amine was also dose-related. Metiamide competitively inhibited the depressor effect of histamine. Prior treatment of the ear vessels with metiamide alone caused an increase in histamine-induced perfusion pressure. From these results it was concluded that the predominant pressor effect of histamine on the vascular bed of the rabbit ear is mediated through the H1-receptors and the depressor effect of the amine through histamine H2-receptors.


Relative pre- and postsynaptic potencies of alpha-adrenoceptor agonists in the rabbit pulmonary artery. The rabbit pulmonary artery contains postsynaptic alpha-adrenoceptors which meidate smooth muscle contraction; its noradrenergic nerves contain presynaptic alpha-adrenoceptors which mediate inhibition of the release of the transmitter evoked by nerve impulses. Dose-response curves for the pre- and postsynaptic effects of eight alpha-receptor agonists were determined on superfused strips of the artery in the presence of cocaine, corticosterone and propranolo. 1. According to the concentrations which caused 20% of the maximal contraction (EC20 post), the postsynaptic rank order of potency was: adrenaline greater than noradrenaline greater than oxymetazoline greater than naphazoline greater than phenylephrine greater than tramazoline greater than alpha-methylnoradrenaline greater than methoxamine. The pA2 values of phentolamine againstoxymethazoline, phenylephrine, alpha-methylnoradrenaline and methoxamine were 7.43, 7.48, 7.59 and 7.69, respectively. 2. For the investigation of presynaptic effects, the arteries were preincubated with 3H-noradrenaline. All agonists inhibited the overflow of tritium evoked by transmural sympathetic nerve stimulation. According to the concentrations which reduced the stimulation-induced overflow by 20% (EC20 pre), the rank order of potency was: adrenaline greater than oxymetazoline greater than tramazoline greater than alpha-methylnoradrenaline greater than noradrenaline greater than naphazoline greater than phenylephrine greater than methoxamine. 10(-5) M phentolamine shifted the presynaptic dose-response curves for moradrenaline and oxymethazoline to the right. 3. The ratio EC20 pre/EC20 post was calculated for each agonist as an index of its relative post- and presynaptic potency. According to the ratios, the agonists were arbitrarily classified into three groups. Group 1 (ratio about 30: preferentially postsynaptic agonists) comprised methoxamine and phenylephrine; group 2 (ratio near 1; similar pre- and postsynaptic potencies) comprised noradrenaline, adrenaline and naphazoline; group 3 (ratio below 0.2; preferentially presynaptic agonists) comprised oxymetazoline, alpha-methylnoradrenaline and tramazoline (as well as clonidine). 4. Preferentially presynaptic and preferentially postsynaptic agonists had opposite effects on the basoconstrictor response to nerve stimulation. Methoxamine and phenylephrine either did not change or enhanced, but never reduced, the response. In contrast, oxymetazoline, alpha-methylnoradrenaline and tramazoline at low concentrations selectively inhibited the response to stimulation at low frequency (0.25-2Hz). 5. It is concluded that alpha-adrenoceptor agonists vary widely in their relative pre- and postsynaptic potencies, possibly because of structural differences between pre- and postsynaptic alpha-receptors. Pre- and postsynaptic components contribute to their overll postsynaptic effec in actively transmitting synapses. The preferential activation of presynaptic alpha-receptors results in alpha-adrenergic inhibition of synaptic transmission.


Investigation into some imidazoline compounds, with respect to peripheral alpha-adrenoceptor stimulation and depression of cardiovascular centers. Peripheral alpha-adrenoceptor stimulation was tested by means of hypertensive effects of the drugs following i.v. injection in spinal rats. Naphazoline (NP), oxymetazoline (OM), St 91-2-(2,6-diethylphenylimino)-2-imidazolidine--and St 1697--2-(2-ethyl, 6-methylphenylimino)-2-imidazolidine--were 3 to 5 times more potent in tthis respect thatn clonidine (CLON) whereas St 363--2-(2,4-dichlorophenylimino)-2-imidazolidine--and xylazine (XY) exerted only approx. 1/20 the effect of that of clonidine. Sympathoinhibitory activity after i.v. injection was tested by the bradycardiac effect in vagotomized rats; St 1697, St 363 and XY were active, approx. 1/10-1/30 of CLON, whereas NP, OM and St 91 were inactive. However, following intracisternal (i.ci.) injection of cardiovascular depression, typical for clonidine: (1) in dogs with blocked beta-adrenoceptors, the drugs facilitated the vagally meditated cardiodepressor reflex in response to baroreceptor stimulation by i.v. injection of angiotensin; (2) in dogs treated with atropine and in (3) vagotomized cats (only NP, OM and St 363) a long lasting decrease in heart rate was observed. Some of the experiments were complicated by increases in blood pressure, due to the "leakage" of small amounts of the highly vasopressor active drugs, from the cisternal spaces into the peripheral circulation. The majority of results indicated, that the central cardiovascular depressor effects of the tested drugs depend on their alpha-adrenoreceptor stimulating potency and on their ability to penetrate from cerebrospinal fluid or from the blood to cardiovascular centers. Relationships between the ability for penetration and the lipoid affinity are discussed.


The regulation of striatal tyrosine hydroxylase. Effects of gamma hydroxybutric acid and healperidol. Gamma-hydroxybutyric acid (GHBA) in doses that increased the striatal dopamine (DA) content of rat brain failed to increase the affinity of striatal tyrosine hydroxylase (TH) for its pterdine cofactor or to change the sensitivity of the enzyme to the inhibition by DA. Haloperidol (1 mg/kg) decreased the apparent Km of striatal TH for the pteridine cofactor. However, when GHBA was injected before haloperidol it prevented the decrease in the apparent Kn of TH, in a dose related manner. In vitro GHBA (10(-4) M) neither changed the stimulation of the striatal adenylyl cyclase by DA nor its inhibition by haloperidol. These results suggest that in striatal dopaminergic terminals the Kn of TH for the pteridine cofactor is regulated by an molecuular mechanism which requires that the impulse flow in the DA neurons is unimpaired.


Host-tumor relationship XXXIII. Inhibitor of hyaluronidase in blood serum of cancer patients. Inhibiting activity of blood serum was determined from the decrease of N-acetyl-hexosamine end groups of hyaluronic acid products released by testicular hyaluronidase. Maximum inhibition was observed within the region of pH from 6.5 to 6.8. Correlation between the serum concentration and its inhibiting effect on hyaluronidase was found within the range of the final dilution of serum (in the reaction mixture) from 9 to 45 X. Blood sera of cancer patients showed statistically significant increase of hyaluronidase inhibitor as compared with that of health people.