Cellular energy metabolism, trans-plasma and trans-mitochondrial membrane potentials, and pH gradients in mouse neuroblastoma. A method for quantitative evaluation of transmembrane electrical potential and pH gradients across a subcellular compartment in an intact cell is presented. This approach has been applied in studies of mouse neuroblastoma C-1300 clone NB41A3, in which the transmembrane electrical potential and pH gradients and the mitochondrial volume percent have been determined. Membrane potentials and pH gradients were measured by two different methods. Equilibrium distributions of [(3)H]triphenylmethyl phosphonium and [(14)C]-thiocyanate ions gave calculated apparent membrane potentials of -77.0 and -29.6 mV, respectively, at 20-25 degrees C; a value of -60.8 mV was obtained from microelectrode measurements. Equilibrium distributions of weak acids ([(14)C]trimethylacetic acid and 5,5-di[(14)C]methyl-2,4-oxazolidine-dione) and of weak bases ([(14)C]dimethylamine and [(14)C]trimethylamine) gave calculated upper and lower limits of the pH gradient (Delta pH = pH(e) - pH(i)) of -0.14 and -0.21 pH unit, respectively. The microelectrode measurements showed that the intracellular pH is within 0.1 of a pH unit or less of the extracellular pH over the extracellular pH range of 7.35-6.85. The mitochondrial volume percent calculated on the basis of the measured cytochrome c content is 5.6 +/- 1.2% and compares well with estimates of 5.4 +/- 1.1% obtained from 25 electron micrographs. Measurements of the cellular energetic parameters gave values within the range found in other cells and perfused organs. Comparison of the results of the microelectrode and equilibrium measurements permits estimates of the electrical potential and pH gradients across the mitochondrial membrane (mitochondria-to-cytoplasm gradients) to be made and suggests that the trans-mitochondrial membrane protonmotive force in the intact cell cannot be greater than -143 mV.


Translocation of glutathione from lymphoid cells that have markedly different gamma-glutamyl transpeptidase activities. Translocation of intracellular glutathione to the medium was studied in lymphoid cells (grown in tissue culture) that have very high, very low, or intermediate levels of membrane-bound gamma-glutamyl transpeptidase, in the absence and presence of various inhibitors of this enzyme. The data show that glutathione is translocated to the medium by all of the cell lines studied, but that glutathione does not accumulate in the medium unless the cellular transpeptidase activity is either very low or substantially inhibited. Translocation of glutathione does not seem to be directly related to the activity of gamma-glutamyl transpeptidase. The present and previous [Griffith, O.W. & Meister, A. (1979) Proc. Natl. Acad. Sci. USA 76, 268--272] findings suggest that translocation of intracellular glutathione is a general property of many mammalian cells. Glutathione exported from cells that have membrane-bound transpeptidase may be recovered by the cell in the form of transpeptidation or degradation products. Translocation of glutathione may also reflect operation of a rather general mechanism that protects and maintains the integrity of cell membranes.


Antibodies raised against purified beta-adrenergic receptors specifically bind beta-adrenergic ligands. Antibodies raised against purified beta-adrenergic receptors themselves specifically bind beta-adrenergic ligands. Digitonin-solubilized frog (Rana pipiens) erythrocyte beta-adrenergic receptors, purified 100- to 200-fold by adsorption to an alprenolol-agarose affinity support and specifically eluted from the affinity resin by 1-100 mM (+/-)-isoproterenol, were used to immunize six rabbits. All immune sera, in contrast to preimmune sera, bound the beta-adrenergic antagonist [(3)H]Dihydroalprenolol binding activity was due to immunoglobulins. By competition studies, antibody [(3)H]dihydroalprenolol binding was found to display a specificity and stereoselectivity resembling that of the beta-adrenergic receptor, [i.e., (-)-isoproterenol > (-)-epinephrine > (-)-norepinephrine; alprenolol approximately propranolol >> phentolamine = aloperidol; and (-) isomers of both agonists and antagonists 10-100 times more potent than (+) isomers]. A portion of the [(3)H]dihydroalprenolol binding antibodies could be specifically adsorbed onto purified frog erythrocyte membranes, whereas Xenopus and human erythrocyte membranes, both of which are almost devoid of beta-adrenergic receptors, were ineffective in adsorbing [(3)H]dihydroalprenolol binding antibodies. We suggest that the likely immunogen was a beta-adrenergic receptor-isoproterenol complex and that immunization with drugs noncovalently bound to their receptors might be a means of raising antibodies to biologically active otherwise nonimmunogenic small molecules. Such antibodies, whose specificity mimics that of a receptor, should also provide useful models for the study of the structure of the receptor binding sites.


Genetic variation and relative catalytic efficiencies: lactate dehydrogenase B allozymes of Fundulus heteroclitus. In order to evaluate whether functional differences exist between allelic variants of a B type lactate dehydrogenase (LDH; L-lactate:NAD+ oxidoreductase, EC 1.1.1.27) in the teleost fish Fundulus heteroclitus (Linnaeus), the kinetic properties of pyruvate reduction were examined. While the pH dependence and the temperature dependence for maximal catalysis were indistinguishable among the allozymes, reaction velocities at low pyruvate concentrations were significantly different. At pH values below 8.00, the LDH-BbBb allozyme showed a greater reaction rate at lower temperatures (e.g., 10 degrees C) than LDH-BaBa. The phenomenon was reversed at higher temperatures (e.g., greater than 25 degrees C) for pH values between 6.50 and 7.00. The rates for the heterozygous phenotype, LDH-BaBb, were not the arithmetic average of the two homotetrameric allozymes. When reaction rates were compared at constant relative alkalinity, that is, a constant [OH-]/[H+] ratio, the findings were similar. The differences in the temperature dependence and the pH dependence for pyruvate reduction found between the LDH-B allozymes may reflect a selective adaptation and help explain the geographical variation in the Ldh-B gene frequencies of F. heteroclitus.


From extracellular to intracellular: the establishment of a symbiosis. The colonization of host cells by modern symbionts is surveyed. The morphological distinction between extracellular and intracellular symbionts is not sharp, and the various kinds of association can be arranged in a graded series of increasing morphological integration of the symbiont into the host cell. Apart from some aggressive parasitic infections, the great majority of symbionts are enclosed by a host membrane in a vacuole. Those not enclosed in a host vacuole usually cannot be cultivated outside the cell. It is therefore surmised that encirclement by a vacuolar membrane would only disappear, if at all, in the later stages of the evolution of intracellular symbiosis. Recognition mechanisms between host and symbiont occur, but have been little studied. In some associations, recognition at surface contact occurs, and there is evidence for the involvement of lectins in certain cases. In other associations, recognition may occur wholly or in part after the entry of symbiont into host cells. After entry, special mechanisms for the biotrophic transfer of nutrients from symbiont to host develop. Both the symbiont population size and its rate of increase are strictly regulated by the host cell; symbiont metabolism may be controlled likewise. Rates of evolution of intracellular symbionts are probably very rapid, owing in part to responses of the host cell to its symbiont.


Regulation of numbers of intracellular algae. Members of three classes of unicellular algae have exploited an intracellular habitat and occur as endosymbionts in aquatic invertebrates, including Protozoa. Such associations manifest a range of host--symbiont cellular interactions and achieve stability through the regulation of symbiont numbers. The mechanism of regulation is poorly understood. Steady-state algae:host cell ratios might be achieved by expulsion, digestion, or inhibition of growth of algal symbionts. Digestion and expulsion have been observed directly in some associations but their role in regulating numbers is circumstantial. Inhibition of growth as a result of nutrient limitation or inhibitor secretion is an attractive, but inadequately tested, hypothesis. The relation between the host cell mitosis and algal proliferation is a potential focal point for further study.


The interactions of intracellular Protista and their host cells, with special reference to heterotrophic organisms. Intracellular genera are found in all the major groups of Protista, but are particularly common among the dinoflagellates, trypanosomatid zooflagellates and suctorian ciliates; the Sporozoa are nearly all intracellular at some stage of their life, and the Microspora entirely so. Intracellular forms can dwell in the nucleus, within phagosomal or other vacuoles or may lie free in the hyaloplasm of their host cells. Organisms tend to select their hosts from a restricted taxonomic range although there are some notable exceptions. There is also great variation in the types of host cell inhabited. There are various reasons for both host and cell selectivity including recognition phenomena at the cell surfaces. Invasion of host cells is usually preceded by surface interactions with the invader. Some organisms depend upon phagocytosis for entry, but others induce host cells to engulf them by non-phagocytic means or invade by microinjection through the host plasma membrane. Protista avoid lysosomal destruction by their resistance to enzyme attack, by surrounding themselves with lysosome-inhibiting vacuoles, by escaping from the phagosomal system into the hyaloplasm and by choosing host cells which lack lysosomes. Nutrition of intracellular heterotrophic organisms involves some degree of competition with the host cell's metabolism as well as erosion of host cell cytoplasm. In Plasmodium infections, red cells are made more permeable to required nutrients by the action of the parasite on the host cell membrane. The parasite is often dependent upon the host cell for complex nutrients which it cannot synthesize for itself. Intracellular forms often profoundly modify the structure and metabolism of the host cell or interfere with its growth and multiplication. This may result in the final lysis of the host cell at the end of the intracellular phase or before the infection of other cells. Certain types of intracellular organisms may have arisen initially as forms attached to the cell surface of digestive or other organs, but the intracellular habit appears to have arisen independently in several groups of Protista.


From extracellular to intracellular: the establishment of mitochondria and chloroplasts. Paracoccus and Rhodopseudomonas are unusual among bacteria in having a majority of the biochemical features of mitochondria; blue-green algae have many of the features of chloroplasts. The theory of serial endosymbiosis proposes that a primitive eukaryote successively took up bacteria and blue-green algae to yield mitochondria and chloroplasts respectively. Possible characteristics of transitional forms are indicated both by the primitive amoeba, Pelomyxa, which lacks mitochondria but contains a permanent population of endosymbiotic bacteria, and by several anomalous eukaryotic algae, e.g. Cyanophora, which contain cyanelles instead of chloroplasts. Blue-green algae appear to be obvious precursors of red algal chloroplasts but the ancestry of other chloroplasts is less certain, though the epizoic symbiont, Prochloron, may resemble the ancestral green algal chloroplast. We speculate that the chloroplasts of the remaining algae may have been a eukaryotic origin. The evolution or organelles from endosymbiotic precursors would involve their integration with the host cell biochemically, structurally and numerically.


Possible evolutionary significance of spirochaetes. Large symbiotic spirochaetes of the family Pillotaceae (e.g. pillotinas) are found in dry wood and subterranean termites (Hollande & Garagozlou 1967). These morphologically distinctive spirochaetes comprise several genera. Some of them contain microtubules within their protoplasmic cylinders. They demonstrate a variety of relations with their termite and protist hosts. Some are free-living within the lumen of the intestine, some tend to be associated with filamentous and other bacteria, some are found regularly coursing between the numerous undulipodia ( = eukaryotic flagella, cilia, and other (9 + 2) organelles of motility) of hypermastigotes and polymastigotes. Still other smaller termite spirochaetes are regularly attached to protists via specialized attachment sites. Some even form motility symbiosis with their host protists. The analogy between the behaviour of host-associated spirochaetes and the possible steps in the origin of the undulipodia and mitotic system of eukaryotes is discussed briefly.


The cell as an extreme environment.

The cell as an extreme environment. Living cells and their intracellular parasites show many of the characteristics ascribed to extreme environments and their dominant species. The diversity of species colonizing intracellular habitats is low, and successful inhabitants exhibit special fitness traits that often render them obligately dependent on residence within a host cell. However, the diversity-limiting factor in the extreme environment of the host cell interior is not abiotic, as it is in conventional extreme environments. It is biotic: the living cell itself and its many activities. Host cells bar the entrance to most would-be parasites, they destroy most of those that do manage to get inside, and they deny parasites free access to many components of their soluble metabolite pools. Successful intracellular parasites have evolved fitness traits that give them the capacity to survive in the face of diversity-limiting factors or to modify the intracellular habitat so that those factors no longer operate. Looking on the cell as an extreme habitat emphasizes its simultaneous roles as environment, antagonist, and competitor.


Interactions between Bdellovibrio and its host cell. The bdellovibrios are extremely small bacteria with the unique property of being parasites of other (gram-negative) bacteria. In the presence of viable and susceptible bacteria a Bdellovibrio cell physically 'attacks' an individual host cell, attaches to its surface, penetrates the cell wall, and multiples within the periplasmic (intramural) space of its prey. The invading Bdellovibrio and its progeny degrade and consume the cellular constituents of the invaded host bacterium. This process finally results in complete lysis of the host cell and release of the Bdellovibrio progeny. From a population of parasitic bdellovibrios, derivatives can be selected that grow on complex nutrient media. Currently, none of the different nutritional types can be propagated in a fully defined synthetic medium. By degradation of the cellular constituents of the host the Bdellovibrio cell in its periplasmic space has available all the monomeric subunits needed to synthesis of the macromolecules. Peculiarities of Bdellovibrio metabolism with respect to uptake of preformed molecules and energy efficiency are discussed.


The Rhizobium--legume symbiosis.

The Rhizobium--legume symbiosis. The rhizobia are soil microorganisms that can interact with leguminous plants to form root nodules within which conditions are favourable for bacterial nitrogen fixation. Legumes allow the development of very large rhizobial populations in the vicinity of their roots. Infections and nodule formation require the specific recognition of host and Rhizobium, probably mediated by plant lectins. Penetration of the host by a compatible Rhizobium species usually provokes host root cell division to form the nodule, and a process of differentiation by both partners then ensues. In most cases the rhizobia alter morphologically to form bacteroids, which are usually larger than the free-living bacteria and have altered cell walls. At all stages during infection, the bacteria are bounded by host cell plasmalemma. The enzyme nitrogenase is synthesized by the bacteria and, if leghaemoglobin is present, nitrogen fixation will occur. Leghaemoglobin is a product of the symbiotic interaction, since the globin is produced by the plant while the haem is synthesized by the bacteria. In the intracellular habitat the bacteria are dependent upon the plant for supplies of energy and the bacteroids, in particular, appear to differentiate so that they are no longer able to utilize the nitrogen that they fix. Regulation of the supply of carbohydrate and the use of the fixed nitrogen thus appear to be largely governed by the host.


'Cells' and 'organisms' as a habitat for DNA. Although the bulk of the hereditary information in bacteria is organized as a single chromosome, it has been known for some years now that bacteria may also carry pieces of self-replicating extrachromosomal DNA. These units are known as plasmids. Sometimes such plasmids carry the information necessary to give rise to mature bacterial viruses under appropriate conditions, but in other cases they specify the production of enzymes and other proteins which alter the bacterial phenotype. Plasmids are often inessential for survival of bacteria, although they may widen the range of environmental conditions under which they flourish. Thus plasmids may be thought of as adventitious additions to the genetic content of bacterial cells. Recently it has become clear that furthur organizational units of DNA are to be found in bacterial cells. These units are called insertion sequences and transposons. Unlike plasmids and the chromosome, however, these DNA units do not carry enough genetic information to specify their own independent replication: they must rely on plasmids or the chromosome for that purpose. Nevertheless they behave in many respects as independent functional units. Although it is possible to think of the chromosome, plasmids and transposons/insertion sequences as three distinct hierarchies of bacterial DNA, genes may move from one hierarchy to another; and such transitions have important implications for the evolution of bacterial populations. Moreover, their study in bacteria may throw much light on the type of DNA interactions occurring in higher cells.


Interactions and DNA transfer between Agrobacterium tumefaciens, the Ti-plasmid and the plant host. Agrobacterium tumefaciens is a gram-negative bacterium with the unique capacity to induce neoplasmic transformations in dicotyledonous plants. Recently, both the mechanism and the biological significance of this transformation have been elucidated. Agrobacterium tumefaciens strains contain a large extrachromosomal DNA plasmid (the Ti-plasmid). This Ti-plasmid is responsible for the oncogenic properties of Agrobacterium strains. A particular segment of the Ti-plasmid, containing information determining the tumorous growth pattern and the synthesis of so-called 'opines', e.g. octopine (N-alpha-(D-1-carboxyethyl)-L-arginine) and nopaline (N-alpha-(1,3-dicarboxypropyl)-L-argine), is transferred and stably maintained and expressed in the transformed plant cells. This phenomenon can be understood as a 'genetic colonization' of the plant cells by bacterial plasmid DNA so that the transformed plant cells will produce and secrete into the medium amino acid derivatives (the opines) that Ti-plasmid carrying agrobacteria can selectively use as carbon and nitrogen sources.


Symbionticism revisited: a discussion of the evolutionary impact of intracellular symbioses. Wallin (1927) first published the notion that the fusion of bacteria with host cells was the principal source of genetic novelty for speciation. He suggested that mitochondria are transitional elements in this process. While the significance that he attributed to symbiosis now seem excessive, he was one of the first authors to be aware of the evolutionary potential of symbiotic events and his view of mitochondria may not seem strange to many cell biologist today. The most significant evolutionary development which has been attributed to intracellular symbiosis is the origin of eukaryotic cellular organization. The current status of the 'serial endosymbiosis hypothesis' is briefly review. The case for the symbiotic origin of the chloroplast, based principally on 16 S RNA oligonucleotide cataloguing, is very strong. Mitochondrial origins are more obscure but also appear to be symbiotic due to recent 18 S cataloguing from wheat embryos. The probablility of the multiple origin of some eukaryotic organelles is also examined, the processes in question being the acquisition of distinct stocks of chloroplasts from disparate photosynthetic prokaryotes and the secondary donation of organelles from degenerate eukaryotic endosymbionts to their hosts, with specific reference to the dinoflagellates Peridinium balticum, Kryptoperidinium foliaceum and the ciliate Mesodinium rubrum. It is concluded that the evolutionary potential of intracellular symbiosis ('cytobiosis': a term introduced in this paper) is great, with the best established influence being on the origin of eukaryotic chloroplasts. Together with the potential effects of viral vectors, symbiosis serves as a supplementary speciation mechanism capable of producing directed evolutionary changes. It is likely that these processes will explain some of the apparent anomalies in evolutionary rates and direction which are not readily explicable by the conventional synthetic theory of evolution.


Preservation of platelet viability and function during storage of concentrates. It has now been substantiated that the viability and function of concentrated platelets stored for 72 hours are best preserved at a storage temperature of 22 degrees C rather than 4 degrees C. However, in order to preserve viability at 22 degrees C several other conditions must be met and maintained, namely, an appropriate storage bag (Fenwal's PL-146, McGaw or Cutter's CL-2383), constant gentle agitation with a platform shaker, CPD anticoagulation, and a platelet concentration of less than 1.6 x 10(12)/liter. Unfortunately, in vitro parameters which can accurately assess stored platelet viability and function are not reliable.


Effect of desglycinamide(9)-lysine(8)-vasopressin and prolyl-leucyl-glycinamide on oral ethanol intake in the rat. Rats given ethanol in their drinking water at a concentration that permitted adequate fluid intake gradually accepted higher concentrations and consumed larger amounts of ethanol. These increases were augmented when daily subcutaneous injections of 1 microgram of desglycinamide9-lysine8-vasopressin (DGLVP) or 10 microgram of prolyl-leucyl-glycinamide (PLG) were given concomitantly. Nonsignificant changes in ethanol consumption were seen with injections of 1 microgram PLG, or 0.42 or 42 microgram of lysine8-vasopressin (LVP). In a second experiment 4 microgram DGLVP given every second day as a long-acting zinc phosphate complex, commencing after the increases in ethanol intake had taken place, failed to produce any change in ethanol consumption subsequently. In both Experiments 1 and 2, the rats were switched from forced ethanol intake to a choice between ethanol and tap water. On these tests there was only marginal evidence of peptide-produced changes in ethanol intake.


[Liquid-liquid extraction for the quantitative determination of sorbed drugs in polyethylene. Part 40: Contributions to the problems of the use of plastic containers for liquid dosage forms (author's transl)]. Salicylic acid was used as a model substance to establish the conditions under which liquid-liquid extraction will yield reproducible results when applied in sorption studies. As the low-density polyethylene used is soluble in toluene only at a temperature above 80 degrees C, an extraction apparatus was developed for transferring salicylic acid from the hot lipophil phase to an appropriate aqueous phase. After optimization of the experimental conditions, the recovery rate was 82.6% with a standard deviation of +/- 2.9%. The procedure is suited for routine determinations of water-extractable sorbed substances.


[Biochemical mechanisms for the effect of alcohol on the brain]. This paper reviews and discusses basic knowledge of biochemical mechanisms of action of ethanol upon the central nervous system, the emphasis being on effects upon cerebral membrane structures and processes as well as mechanisms of chemical synaptic transmission. Results of detailed studies into material and steric changes in membranes, ion-dependent adenosine triphosphatases, variations in ion balance, effects upon cyclic nucleotides, influences on special transmitter systems, and mechanisms of formation of morphine-analogous condensation products are presented. In addition, open questions are derived and formulated in problem complexes.


Benztropine prophylaxis of dystonic reactions. The effectiveness of antiparkinson medication for the prevention of drug induced dystonias has remained a question. Forty patients with acute psychosis who received high potency oral antipsychotic drugs were interviewed to determine the incidence of acute dystonia. An eleven-fold increase in dystonia was found in patients who received no prophylactic medication. Such prophylaxis appears effective in preventing acute dystonia.


Environmentally induced differences in susceptibility of rats to CNS stimulants and CNS depressants: evidence against a unitary explanation. It has been suggested that socially isolated rats are more aroused then rats raised in social groups. This hypothesis was tested by examining amphetamine-induced activity and stereotypy in social and isolated rats of both sexes in both the active and inactive phases of their diurnal activity cycle. In socially raised rats it was possible to produce behavioural profiles similar to those of isolated rats by increasing the arousal level of the social rat. However, the complex interaction of housing conditions, diurnal variation and gender with drug dose suggests that one intervening variable such as arousal is too simplistic an explanation. In subsequent experiments, stereotypy was enhanced by a familiar environment, and there was a clear dissociation between the effects of CNS stimulants and CNS depressants. The increased susceptibility of isolates to CNS stimulants depends on social isolation for a short period before 45 days of age; the decreased susceptibility of isolates to CNS depressants may be produced by isolation at any age. We conclude that there is no evidence that isolated rats are hyperaroused.


The in vitro binding of 2,2', 5,5'-tetrachlorobiphenyl metabolites to rat liver microsomal proteins. The in vitro association of tritium labeled 2,2', 5,5'-tetrachlorobiphenyl (TCB) with the microsomal fraction isolated from rat livers has been investigated in a metabolizing system. It was found that the control microsomes, capable of only minimal TCB metabolism, had 92% of the total microsomal radioactivity associated with lipids, while only 61% was associated with the phenobarbital induced microsomal lipid. The radioactivity per mg microsomal protein was the same for both induced and noninduced microsomes; however, very important qualitative differences were found. Only the proteins of the induced system contained a protein (s) (MW = 45,000 g/mole) capable of specifically binding a TCB metabolite. This binding required metabolism and was TCB concentration dependent. The specificity of this association was confirmed by dialysis and this data could be analyzed by the Scatchard-Klotz equation. These calculations allowed the evaluation of the number of binding sites (38 micron moles/g of total microsomal protein) and the apparent binding constant (1.4 x 10(7) l/mole). These data are consistent with strong noncovalent interaction of a 2,2', 5,5'-TCB metabolites, but do not exclude the possibility of covalent binding of other non-dialyzable metabolites.


Inhibition of cardiac protein synthesis by prolonged ethanol administration. Effects of prolonged ethanol consumption have been studied on the rates of cardiac protein synthesis. Prolonged ethanol consumption resulted in a significant decrease in cardiac contents of total protiens and RNA. Chronic exposure to ethanol did not result in an alteration in cardiac DNA content. The rates of protein synthesis measured by determining the rates of (U-14C)-leucine incorporation into cardiac proteins showed that chronic ethanol-feeding leads to a significant inhibition of protein synthesis. Studies with ribosomes and pH 5 enzyme fractions of heart showed that prolonged ethanol consumption inhibits the capacity of both these fractions to synthesize proteins. Acute administration of ethanol or in vitro addition of ethanol does not affect the cardiac protein synthesis in the heart. The acetaldehyde-mediated inhibition of cardiac protein synthesis can be partially prevented by antabuse. These observations suggest that, at least some of the deleterious effects of chronic ethanol consumption on the heart may be exerted through the inhibition of protein synthesis in the cardiac muscle.


[Endoscopic diagnosis and therapy of colorectal tumours (author's transl)]. 1. If endoscopy leads to the suspicion of an exulcerated and/or polypous carcinoma of the colon, surgical intervention is the primary therapy. Histological classification of the tumour should be effected by means of endoscopic biopsy. 2. (Familial) adenomatosis of the colon requires colectomy. 3. Broad-based polypi resembling a lawn where a large wound area must be expected by electrocoagulation (risk of perforation), and pediculate polypi of the (rare) size of 3 cm and more (risk of haemorrhage) should not be resected via endoscopy but by surgery. 4. Solitary or multiple polypi of the colon not covered by points 1 to 3 above, are primarily for reasons of diagnosis an indication for endoscopic polypectomy. Biospy in the case of adenomas to clarify the histological structure and to obtain qualitative and quantitative information regarding malignant degeration, must be discouraged (""partial diagnosis''.) 5. Basing on the current state of knowledge it is assumed that effective prohylaxis of cancer is achieved by the endoscopic removal of benign adenomas of the colon. 6. It is also assumed that effective cancer therapy can be realised by the endoscopic removal of adenomas which have already undergone malignant degeration (adenoma with severe cellular atypia, invasive differntiated adenocarcinoma in the head of the adenoma.)


Clinical experiences with beta adrenergic blocking therapy on burned patients. The cachexy which appears in the toxicoseptic phase of burn injuries can be prevented by the regular use of beta-adrenergic blocking drugs. This therapy is based upon data which maintain that the burned organism passes through a beta-modulated adrenergic receptor state as a result of the endocrine changes taking place between the 3rd and the 5th post-burn day. The author describes his own positive experiences of beta-adrenergic blocking therapy. The possible contra-indications (obstructive lung diseases, shock) to this therapy are discussed. Finally he draws attention to some of the side effects which were observed (E.C.G. conduction disturbances).


N-formyliminodiacetic acid, a new compound from the reaction of nitrilotriacetic acid and chlorine. It has been proposed that nitrilotriacetic acid be substituted for trisodium polyphosphates in detergents as a way to reduce the rate of eutrophication in the Great Lake Basin. The reaction of nitrilotriacetic acid with chlorine-containing solutions produces a hitherto unknown degradation production, N-formyliminodiacetic acid, in high yield. The toxicological and environmental implications of this reaction are unclear.


Neuronal supersensitivity to acetylcholine induced by kindling in the rat hippocampus. Kindling is an experimental model of epilepsy in which periodic brain stimulation induces the progressive development of electrical and behavioral seizures. A kindling-induced electrical seizure (afterdischarge) in the rat hippocampus produces prolonged neuronal supersensitivity to microiontophoretically applied acetylcholine after a latency of 40 to 60 minutes. Neuronal acetylcholine supersensitivity is correlated with the further progression of kindling. A larger hippocampal after-discharge is elicited by a subsequent kindling stimulus delivered in the presence of acetylcholine supersensitivity, but not by one delivered before the onset of the supersensitivity. The results suggest that alteration of synaptic sensitivity to acetylcholine may contribute to kindling and epileptogenesis.


Embryonic development of identified neurons: temporal pattern of morphological and biochemical differentiation. Individually identified neurons can be recognized in grasshopper embryos, and are accessible to examination by morphological, physiological, and biochemical techniques from their birth to their maturation. Only after the axon of an identified neuron reaches its postsynaptic target does the neurotransmitter accumulate, the soma rapidly enlarge, and the central arborizations greatly expand.


Separation of D and L amino acids by liquid chromatography: use of chiral eluants. An aqueous eluant containing a chiral copper-proline complex effects the separation of underivatized amino acid enantiomers on an ion-exchange column. The stereoselectivity is ascribed to differences in stability of the diastereomeric amino acid-copper compexes formed in solution. A simple change in the chirality of the eluant reverses the order of the enantiomer elution. For detection and quantification of picomole amounts of amino acids, the eluant is monitored for fluorescence after reaction with o-phthalaldehyde, a reagent insensitive to proline but highly sensitive for amino acids containing a primary amino group.


Cellular applications of 31P and 13C nuclear magnetic resonance. High-resolution nuclear magnetic resonance (NMR) studies of cells and purified mitochondria are discussed to show the kind of information that can be obtained in vivo. In suspensions of Escherichia coli both phosphorus-31 and carbon-13 NMR studies of glycolysis and bioenergetics are presented. In rat liver cells the pathways of gluconeogenesis from carbon-13-labeled glycerol are followed by carbon-13 NMR. In the intact liver cells cytosolic and mitochondrial pH's were separately measured by phosphorus-31 NMR. In purified mitochondria the internal and external concentrations of inorganic phosphate, adenosine diphosphate, and adenosine triphosphate were determined by phosphorus-31 NMR while the pH difference across the membrane was measured simultaneously.


Food deprivation increases oral and intravenous drug intake in rats. Rats given continuous access to etonitazene hydrochloride in their drinking water (5 micrograms per milliliter) more than doubled their drug intake while deprived of food. Another group of rats with implanted jugular catheters self-administered etonitazene (10 micrograms per kilogram) intravenously on a continuous reinforcement schedule, and the number of infusions increased significantly on days when they were deprived of food. These results suggest that feeding condition may be a powerful determinant of drug-reinforced behavior.


Erysipelas-like syndrome caused by Streptococcus pneumoniae. Streptococcus pneumoniae produced an erysipelas-like eruption in a patient with the nephrotic syndrome. The eruption and accompanying fever resolved upon treatment with aqueous penicillin.


Serum gamma-glutamyl transpeptidase: its specificity and clinical value. The clinical import of the serum gamma-glutamyl transpeptidase (GGTP) level was evaluted in 162 prospectively studied patients. GGTP is helpful in determining the origin of alkaline phosphatase (AP); it clearly separates increased AP of bone and placental origin from that of liver origin. The GGTP level closely parallels the AP level in most instances, but it may be more sensitive in detecting liver disease in anicteric patients. The finding of significantly increased GGTP in patients with chronic aggressive hepatitis as compared to normal levels found in chronic persistent hepatitis may provide a prognostic clue in cases of unresolved hepatitis. The apparent specificity and sensitivity of the GGTP test, combined with ease of performance and low expense, make it a valuable addition to the evaluation of a patient with hepatic disease.


Faecal pH value and its modification by dietary means in South African black and white schoolchildren. Mean faecal pH values did not differ significantly in groups of rural South African Black schoolchildren of 10--12 years who ate their traditional high-fibre low-fat diet, and urban dwellers who consumed a partially westernized diet. However, both means were significantly lower than those of groups of White schoolchildren. In feeding studies of 5 days' duration, mean faecal pH value of Black children became significantly less acid when white bread replaced maize meal, and became significantly more acid when a supplement of 6 oranges was consumed daily. Supplements which consisted of skim milk, butter, and sugar had no significant effect on mean faecal pH value. In White children in an institution, the mean pH value of faeces became significantly more acid when a supplement of 6 oranges, although not of bran 'crunchies', was consumed daily.


The effect of exposure to halogenated anaesthetics on liver glutathione levels in rats. An index of hepatotoxicity. Having studied anaesthetic drug interactions in rats, we report the effects of halogenated anaesthetics on the liver glutathione levels and histology, as well as the results of the enhancement of these effects by microsomal enzyme induction. The anaesthetic agents studied included methoxyflurane, halothane, ethrane, chloroform and fluroxene. While exposure of rats to methoxyflurane, helothane and ethrane produced no significant changes in hepatic glutathione levels, or in liver histology, exposure to chloroform and fluroxene produced marked depression of liver glutathione, especially after microsomal enzyme induction. Furthermore, rats exposed to thses agents after enzyme induction developed gross centrilobular necrosis and died. It is suggested that the study of the effects of any new anaesthetic agent on liver glutathione levels could be a valuable screening test of its hepatotoxic potential, before its clinical trial.


Peptones and mycological reproducibility. The pH reactions, ultraviolet spectra and phosphorus content of solutions of a variety of commercially available peptones all indicated, predictably, considerable differences in the chemical composition of the peptones. The effects of these differences on the outcome of experiments with Candida albicans grown in different peptone media was investigated. The fungus produced germ tubes equally effectively on all such media provided that the inoculum was kept to 10(6) blastospores/ml or less. However, expression of inducible enzyme activities in C. albicans varied extensively from peptone to peptone; there was, for example, an inverse relationship between the inorganic phosphorus content of peptones and the amount of acid phosphomonoesterase detectable in intact blastospores. The results indicated that use of different peptones in "Sabouraud's" media by different laboratories may account for some, but not all, published instances of irreproducibility of experiments with C. albicans.


Orchidopexy in cryptorchidism assessed by clinical, histologic and sperm examinations. A study was undertaken of 187 patients with unilateral and 41 patients with bilateral cryptorchidism before and after orchidopexy. The mean tubular diameter and the mean tubular fertility index were used as quantitative criteria for assessment of the development of the testes. Before orchidopexy, no significant differences were found between scrotal and cryptorchid testes in patients up to six years of age. Thereafter, the scrotal testis showed marked development in distinction to the cryptorchid testis. After orchidopexy, follow-up examinations were carried out when the patients had reached the age, at least, 18 years. Based on the results of the mean tubular diameter and the mean tubular fertility index, a reasonable degree of fertility would be expected in both the unilateral and the bilateral cryptorchid testes. However, histologic assessment of spermatogenesis in the unilateral cryptorchid testes after orchidopexy showed spermatogenic arrest in 81 per cent and in all patients with bilateral cryptorchid testes after orchidopexy. In the unilateral orchidopexy patients, in distinction to the histologic assesment of spermatogenesis, sperm counts gave good results in 80 per cent; evidently, in these patients, the source of the spermatozoa was the contralateral scrotal testis. That spermatogenesis was defective in 20 per cent of the patients after unilateral orchidopexy suggests and underlying systemic factor affecting both testes.


Gastroesophageal reflux and pulmonary aspiration: incidence, functional abnormality, and results of surgical therapy. The incidence of aspiration, the causative esophageal pathophysiology, and the results of surgical therapy were evaluated in 100 patients with abnormal gastroesophageal reflux documented by 24-hour esophageal pH monitoring. Based on historical evidence, 48 patients were suspected to be aspirators. Eight patients had documented episodes of aspiration (drop on esophagela pH, followed by acid taste in mouth and onset of cough or wheezing spell) during the monitoring period. Nine patients were considered to be potential aspirators because they presented oral acid regurgitation without development of pulmonary symptoms. In five patients a primary respiratory disorder (PRD) induced gastroesophageal reflux. The remaining 78 patients had abnormal reflux without aspiartion or regurgitation. Aspirators had a 75% incidence of esophageal motor abnormality on manometry, and the clearance of refluxed acid was significantly delayed in the supine position. A history of heartburn and endoscopic evidence of esophagitis were present in only half of the patients who were documented aspirators. Potential aspirators were spared from aspiration by rapid esophageal clearance of refluxed acid unaffected by changes in body position. Patients with a PRD had higher distal esophageal segment (DES) pressure and normal esophageal motility with minimal esophagitis. Nonaspirators significantly improved their clearance while in the supine position, emphasizing the protective effect of esophageal peristalsis against aspiration. An antireflux procedure in five aspirators raised the DES pressure significantly and returned the reflux status to normal by 24-hour pH-monitoring standards. The incidence of aspiration appears to be less than that suspected by history and is due to a motor disorder that interferes with the ability of the esophagus to clear reflex acid. Abnormal pulmonary symptoms can induce or result from gastroesophageal reflux and, when the latter occurs, an antireflex procedure stops both reflux and aspiration.


Steroid specificity of human placental 5-ene-3 beta-hydroxysteroid oxidoreductase. The properties of 5-ene-3 beta hydroxysteroid oxidoreductase (3 beta-HSD) from human placental homogenates were studied in vitro. The apparent Michaelis constants for 3 beta-HSD with the substrates pregnenolone (delta 5P) and dehydroepiandrosterone (DHA) were 170 nM and nM respectively. The optimal pH for both these substrates was between 10 and 12. With NAD as the substrate, the Km for the pregnenolone was 20 microM and for DHA, 17 microM. The activity of 3 beta-HSD was inhibited by various steroids. Competitive inhibitors (pregnenolone substrate) included: ethynylestradiol (inhibition constant Ki=7.3 nM), DHA (Ki=46 nM), estradiol-17 beta (Ki=46 nM), cholesterol (Ki=0.68 microM) and 16 alpha-hydroxydehydroepiandrosterone (16 alphaOHDHA) (Ki=2.2 microM). When the substrate was DHA, competitive inhibition occurred with the following steroids: ethynylestradiol (Ki=6.4 nM), estradiol-17 beta (Ki=69 nM), pregnenolone (Ki=91 nM), cholesterol (Ki=1.3 microM) and 16 alphaOHDHA (Ki=1.9 microM). 4-Ene-3-ketosteroids such as androstenedione, progesterone (delta 4P), norethindrone and chlormadinone acetate acted as noncompetitive inhibitors towards both substrates.


Suppression of cell-mediated immunity in rats transplanted with kidneys from donors compatible at the major histocompatibility complex. In order to study the immunological status of rats transplanted with H-1-compatible kidney allografts, LEW rats were grafted with F and (Fischer X Lewis)F1 (FLEWF1) kidneys. Most of the F kidneys were rejected within 55 days, only 4 of 24 surviving for more than 4 months. However, two-thirds of the FLEWF1 recipients survived for more than 4 months, the others having died within 64 days. During the first postoperative week, high levels of serum lymphocytotoxic antibodies were found in recipients of F kidneys, and thereafter there was little change. In this respect these rats did not differ from recipients of kidneys with major histocompatibility differences. However, recipients of FLEWF1 kidneys had low haemagglutinating and lymphocytotoxic antibody titres. No serum-blocking factor could be found in kidney of recipients by use of the microcytotoxicity assay (MCA) or inhibition of allorosette formation. Cellular immunity, which was studied by means of the graft-versus-host reaction (GVHR) and the microcytotoxicity assay, was detected in the first postoperative week. This immunity gradually declined, and after 6 weeks it was no longer detectable. The immunological status of the long-term surviving kidney recipients remained unchanged, even when they were provided with further antigenic challenge in the form of two successive donor strain skin grafts.


A factor VIII concentrate of intermediate purity and higher potency. A fractionation method has been modified to produce a factor VIII concentrate of intermediate purity and a potency of about 12 iu/ml, suitable for injection by syringe. The solubility, ease of filtration and purity of the concentrate have been improved by including a stage in which contaminants are precipitated at controlled temperature, pH and ionic strength. A summary of the first 50 production-scale batches is presented. Plasma can be harvested from whole blood and frozen within eight hours of donation, or after overnight storage of whole blood, with no significant difference in overall yield.


[Recurrence of inguinal, undescended, testicle (author's transl)]. An analysis of 94 patients from 1970 to 1977 demonstrates the problems of recurrence of the inguinal undescended testis. In more than 2/3 of the patients operative deficiencies were considered as the cause. The technique of secondary corrective surgery is described. The special difficulties with short internal spermatic vessels are outlined. In 90% of the patients a good anatomical result was obtained but no data are available concerning later functional results.


[Stability of alpha-amylase with immobilization through its different functional groups]. The paper deals with stability of Aspergillus aryzae alpha-amylase immobilized on CM- and AE-celluloses using N,N'-dicyclohexylcarbodiimide by means of binding through its amine or carboxylic groups. The binding of the enzyme with CM-cellulose makes its preparations more stable to the effect of EDTA and elevated temperature (50 degrees C) than under fixation on AE-cellulose.


[Effect of different levels of carbonic acid on activity of dehydrogenases]. A dependence is established for the activity of NADP-dependent isocytrate dehydrogenase and malate dehydrogenase as well as of NAD-malate dehydrogenase on the carbonic acid level. When the biocarbonate concentration in rabbit blood and tissues rise, the activity of cytoplasmic NADP-isocytrate dehydrogenase and NADP-malate dehydrogenase of the liver lowers. The analogous regularity is observed in the in vitro experiments when the bicarbonate concentrations in the medium rises up to 40 mM (within the physiological limits). The activity of NAD-malate dehydrogenase in the soluble fraction of the rabbit liver and kidney increases with the bicarbonate level within the same limits. The found dependence was confirmed in experiments with the enzyme crystalline preparations.


[Use of gastric pH-metry for peroperative evaluation of the completeness of vagotomy]. In 124 patients subjected to various types of vagotomy its completeness was tested peroperatively with Grassi test and intrastomach pH-metry. The results were compared with the Hollander test findings in 112 postoperative cases. It has been ascertained tat the peroperative pH-metry does not allow for judging on the completeness of vagotomy. The postoperative pH-metry tests show only the ability of the stomach mucous membrane to produce hydrochloric acid in response to the maximal histamine stimulation, which can be used for total evaluation of the effectiveness of the operation.


[Effect of benemycin and sodium succinate on metabolism of protons in mitochondria of guinea pig lung and liver tissues]. The data of studies on proton metabolism in mitochondria of the guinea pig lungs and liver with application of benemycin and sodium succinate show that benemycin does not affect the metabolism of protons in the lungs and liver mitochondria. Sodium succinate activates the release of protons from the lungs and liver mitochondria in state 4 according to Chance. When using sodium succinate for reducing the effect of benemycin on theorganism it is found that sodium succinate inhibits the utilization of protons of the lungs and liver mitochondria in state 3 according to Chance and activates this release in state 4, thus favouring an increase in their concentration in the extra-mitochondrial space. Sodium succinate increases survival rate of animals which were administered benemycin to.


[Oxidation of fatty-aromatic aldehydes in liver tissues]. A protein fraction, which did not contain NADP [or NADPH]-dependent aldehyde reductase as well as NAD [or NADP]-dependent aldehyde dehydrogenases, but which catalyzed oxidation of fatty-aromatic aldehydes, was isolated from extract of rat liver tissue using ammonium sulfate fractionation combined with gradient syvorptive chromatography on DEAE-Sephadex A-25 [or Molselect DEAE-25], CM-Sephadex C-25 and gel-filtration on Sephadex G-200. Investigations of molecular weight and catalytic properties of the protein fraction obtained enabled to identify it with xanthine oxidase [EC 1.2.3.2]. Aldehyde dehydrogenases as well as xanthine oxidase are involved in oxidation of fatty-aromatic aldehydes to corresponding fatty acids, besides the reduction of the aldehydes to alcohols, catalyzed by aldehyde reductase and alcohol dehydrogenases.


[Investigations on the pathogenesis of distal renal tubular acidosis (author's transl)]. In distal (type 1) RTA, renal acid excretion is impaired by the inability to establish adequate pH gradients between plasma and distal tubular fluid at any level of acidosis. Main clinical signs in infancy are anorexia, vomiting and failure to thrive. Despite low serum bicarbonate levels the renal threshold of bicarbonate is normal, while urinary pH levels are high even with values below the threshold. Under conditions of bicarbonate-induced systemic alkalosis urinary the pCO2 exceeds blood pCO2 in normal subjects. by contrast, the urinary pCO2 tension is not significantly greater in distal RTA, indicating a failure of the cells of the distal nephron to secrete hydrogen ions even without a gradient. Red cell carbonic anhydrase is within the normal range, whilst the inhibition of carbonic anhydrase activity has no effect on distal tubular function. Until now no histological or enzymatic defect could be detected to explain the ineffective acidification. Bicarbonate loading is followed by a lowering of calcium excretion to within the normal range and a decrease in the uncharacteristic renal hyperaminoaciduria.


Deranged tyrosine metabolism in cirrhosis. In normal individuals, the main route for tyrosine degradation is the hepatic pathway tyrosine→4-hydroxyphenylpyruvic acid→homogentisic acid→CO(2). Quantitatively minor pathways, in large part extrahepatic, are: tyrosine→tyramine→octopamine and tyrosine→dopa→catecholamines.In cirrhosis, the main hepatic pathway is blocked to varying degrees at the first three stages. This appears to be due to lack of activity of the enzymes tyrosine transaminase, PHPA oxidase, and HGA oxidase, the first step being rate limiting. Hypertyrosinemia and tyrosine intolerance result.With the main hepatic pathway partially blocked, an abnormally large amount of tyrosine passes into the normally minor extrahepatic pathway leading to the false neurotransmitters tyramine and octopamine. Overproduction of these amines ensues and they accumulate in the body fluid.The false neurotransmitters can displace catecholamines from their storage sites in the peripheral and central nervous system, and thereby disrupt adrenergic processes in arterioles, kidneys, and brain. Their accumulation in cirrhotic patients may play a role in the pathogenesis of hepatic encephalopathy, hepatorenal syndrome, and hyperdynamic circulation.


[Clinical picture and therapy of sleep in aged, internal-medicine patients]. The management of sleep disorders in elderly patients with internal diseases consists in the first line in rectifying pathophysiological disregulations. Only in the second line, proper hypnotics are to be prescribed. When considered as indispensable, these medicaments are selected according to their toxicity and side effects. In present time, Benzodiazepines are definitely preferred, whereas neuroleptic, anti-depressant and the older drugs are to be taken secondarly in account.


Absence of microsomal lipid peroxidation in acatalasemic mice. Catalase (E.C. 1.11.1.6) activity and NADPH-dependent lipid peroxidation have been measured in liver microsomes from normal and acatalasemic mice. The absence of lipid peroxidation in acatalatic microsomes is not restituted by exogenous catalase as is microsomal methanol oxidation nor is it inhibited by sodium azide, thus suggesting an additional abnormality in these mice.


[Use of sodium nucleinate in the therapy of schizophrenia]. A total of 102 patients with progressive forms of schizophrenia with a pronounced deficital symptomatology were treated. The dynamics of some indices of nonspecific reactivity (properdin, lysozyme, complement) were studied parallely. Yeast sodium nucleinate promotes a softening and reverse development of some deficital symptoms (mainly in an emotional-volutional sphere), decreasing the threshold of sensitivity to neuroleptics. When remission occurred, the immunological indices returned to normal.


On the deacetylase activity of Vi bacteriophage III particles. 1. Using the complete phage particles as an enzyme, O-acetyl (1 leads to 4)-alpha-D-galacturonan (acetylated pectic acid) as a substrate, and gas-liquid-chromatography for the determination of the acid liberated, the virus-catalysed deacetylation of the polymer was studied. The activity was found to be stable up to about 50 degrees C, and from pH 4.5 to 9, with an optimum at pH 7.8; it was not affected by EDTA, or by 1,10-phenanthroline. The initial reaction velocity (at 37 degrees C) exhibited a simple hyperbolical dependence on the substrate concentration, with Km = 10.5 mM for O-acetyl (independent of virus concentration), and Vmax = 15 nmoles/min and 10(10) plaque forming units. The reaction was, however, rapidly inhibited by a partially deacetylated product (but neither by acetate, nor by pectic acid itself). 2. Using the natural substrate, acetylated (1 leads to 4)-2 amino-2-deoxy-alpha-D-galacturonan (Vi polysaccharide, Vi antigen), and a variety of structural analogues, the following conclusions about the substrate specificity of the Vi phage III deacetylase (acetyl-alpha-1,4-galacturonan acylhydrolase) were reached: (a) acetylated galacturonan is as good a substrate as acetylated aminogalacturonan; (b) of the two substrate diastereomers, acetylated alpha-L-guluronan (also 1 ax leads to 4 ax-linked units, but with axial acetyl residues at C-3), and beta-D-mannuronan (1 eq leads to 4 eq-linkages, and axial acetyl groups at C-2), only the former was acted upon, possibly indicating a specificity for the conformation of the polymer rather than for the configuration of the single residues; (c) all acyl analogues tested, O-monofluoroacetyl, O-propionyl, and O-butyryl galacturonan, were inert, showing a high degree of specificity for O-acetyl; (d) the oligomers, acetylated tri- and digalacturonic acid, as well as methyl-alpha-D-galacturonide, were still deacetylated, although more slowly, demonstrating tolerance of the enzyme of substrate size.


[Biochemical changes in guinea pigs after the application of high doses of ascorbic acid]. The effect of high doses of ascorbic acid (AA) (200 mg/kg body weight, s. c.) on the values of some biochemical parameters has been studied in guinea pigs (glucose, lactic acid, SH substances and GSH in blood, proteins and urea, and the LDH, MDH, ASAT, ALAT, and gamma-GT activities in the serum, the acid phosphatase activity in the liver, the gamma-GT activities in the liver and the kidney, and on the levels of SH substances and GSH in various organs). Ascorbic acid was administered to the animals in a single dose or in dialy doses for different periods (4 days, 2 weeks, and 7 weeks). The majority of the studied parameters showed a transient increase or decrease of the values which, however, after long term AA administration mostly returned to normal values. After 7 weeks, the level of urea in the serum remained increased (by approximately 30%) contrary to the decreased level of lactic acid, the gamma-GT activity in the serum, and the GSH level in some organs. In the experimental series carried out in the different seasons of the year, the results differed in some cases especially after short term AA administration. However, in a few cases, even the values obtained from the control animals tended to differ. If similar changes are to be expected in man after AA administration, then on the basis of our results it can be assumed that in the indicated cases the therapeutic long-term application of large AA doses does not present any hazard to the patient.


Fetal breathing movements and maternal exercise. Earlier statements that fetal breathing movements (FBM) are sensitive to changes in the fetal homeostasis prompted the study of the effect of maternal exercise on FBM and fetal heart rate. Forty women in the last trimester of gestation were subjected to a work load (80 W) for 5 min on a bed ergometer cycle; in 30 of them FBM were recorded by A-mode ultrasound, and in 10, the fetal heart rate was monitored by continuous ultrasound. Maternal blood pressure, pulse rate, blood pH and pCO2, and transcutaneous pO2 were also followed. The FBM showed a transient marked increase in incidence immediately after the end of the exercise. No changes in basal level or in baseline variability of the fetal heart rate were found in the recovery period after work. Some possible causes of the observed FBM alterations are discussed. The findings imply that, after this particular form of stress, FBM are a more sensitive indicator of the physiological state of the fetus than the fetal heart rate.


Inhibition and facilitation of lacrimal flow by beta-adrenergic drugs. The effects of adrenergic beta-stimulating and beta-blocking compounds on tear flow were studied in conscious rabbits with a modified Schirmer technique. The tear flow was significantly increased by the unselective beta-adrenergic agonist isoprenaline and by the selective beta1-agonist H 80/62 (racemic Prenalterol, Hässle, Sweden). The effects of the agonists were blocked by the unselective beta-adrenergic antagonist propranolol (Inderal) and by the selective beta1-antagonist metoprolol (Seloken, Lopressor). The experimental data favoured the hypothesis that the beta-receptors involved in the regulation of the tear flow were of the beta1-type. The anatomical location of these beta-receptors is at present unclear but in vitro experiments performed on the rabbit lacrimal glands indicated that the receptors were probably not located on cholinergic nerve-endings as the release of acetylcholine was not influenced by isoprenaline.


Effect of plasma H+-ion concentration on pancreatic HCO-3 secretion. The relationship between the rate of pancreatic HCO-3 secretion and plasma H+-ion concentration was investigated in 15 pentothal anesthetized, secretin infused pigs (1.8 C.U./kg b.w. h-1, intravenously) during acute metabolic and respiratory acid-base disturbances. Pancreatic HCO-3 secretion increasd to 196 +/- 10% of control during alkalosis and fell to 41 +/- 4% of control during acidosis. Partial metabolic compensation of respiratory acidosis restored HCO-3 secretion to 87 +/- 6% of control. A proportional relationship was found between HCO-3 secretion and plasma pH. Different, proportional relationships were found between HCO-3 secretion and plasma HCO-3 concentration during metabolic and respiratory acid-base changes. HCO-3 secretion was independent of H+-ion concentration in pancreatic juice. Plasma H+-ion concentration, therefore, seems to determine the rate of pancreatic HCO-3 secretion. This finding supports the hypothesis that a proton pump is responsible for pancreatic HCO-3 secretion.


Tricyclic antidepressant and cardiovascular drug interactions. Tricyclic antidepressants have anticholinergic, adrenolytic and quinidine-like activity. These actions result in a variety of cardiac and blood pressure effects. Tricyclic antidepressants can reverse the antihypertensive effect of guanethidine and clonidine. Orthostatic hypotension may be increased with diuretics and hydralazine. Myocardial depression may occur with lidocaine, phenytoin or propranolol. Dangerous additive effects may result from concomitant use of a tricyclic antidepressant and either quinidine or procainamide.


HL-A antigens in Takayasu's disease.

HL-A antigens in Takayasu's disease. Takayasu's disease is characterized by a "pulseless" condition which most often occurs in young females from Asian or South American areas. The cause of this disease remains obscure. Recently, we encountered monozygotic, Japanese, identical twin sisters, both of whom were diagnosed as having Takayasu's disease. The parents, two sisters, and one brother are healthy. HL-A typing analyses revealed that one haplotype found in the father had passed only to these twins. Such observations led us to search HL-A typing in Takayasu's disease to determine the possible participation of genetic factors in the pathogenesis of this morbid condition. Ten families, including that of our own patient, have been reported in the literature in Japan, as family cases of Takayasu's disease. HL-A typings in A and B locus analyzed in all family members of six families in attempts to find a common haplotype composed of A9, A10, B5, or BW40 in patients with Takayasu's disease, were confirmed statistically (chi 2 = 7.8, 0.01 less than p less than 0.05). In a population study, HL-A typing analyses of 65 patients with Takayasu's disease also revealed a high frequency of HL-A A10 and HL-A B5 with the level of 15.3 and 17.0 in the chi 2-test (p less than 10(-4)), as compared with the frequency in 128 healthy Japanese. These data strongly suggest that a genetic-related factor has to be given serious consideration.


Acute central chest pain in the elderly. A review of 296 consecutive hospital admissions during 1976 with particular reference to the possible role of beta-adrenergic blocking agents in inducing substernal pain. Two hundred and ninety-six patients were admitted to geriatric medical beds in Cardiff in 1976 with acute central chest pain. One hundred and eighty-six (63 per cent) had a confirmed acute myocardial infarction. Of the 37 per cent without evidence of cardiac infarction, 32 per cent were on beta-blocking drugs. The possible role of adrenergic blocking agents in producing acute central chest pain is discussed.


Improvement of the glucose oxidase immunoenzyme technic. Use of a tetrazolium whose formazan is stable without heavey metal chelation. Glucose oxidase immunoenzymatic localization provides a simple way to show antigens in mammalian tissues, with no need for the quenching of endogenous nonspecific staining. The method is useful for the demonstration of many antigens in formalin-fixed, paraffin-embedded tissues. Improvement of this technic by the use of p-nitro blue tetrazolium chloride (NBT) as the disclosing reagent provides a stable, finely grained localization not possible with the previously used thiazolyl blue (MTT). The described modification makes available an ideal immunoenzymatic stain for the study of tissues at the level of the light microscope. This stain is especially useful for the examination of tissues with hemorrhagic or inflammatory lesions, since there is no endogenous background staining. Slides are permanent, and the technic can be used with peroxidase-labeled antibody to localize two antigens in the same tissue.


Alpha1-antitrypsin quantitation and Pi typing of stored postmortem blood. Correlation of necropsy findings with protease inhibitor levels and phenotype is sometimes desirable. This study was designed to determine the feasibility of postmortem protease inhibitor assessment. One hundred fifteen consecutive postmortem samples, stored at -20 C for 24 to 53 months, were analyzed. The time from death to necropsy, storage time, and the pH values of the sera were correlated with alpha1-antitrypsin levels, trypsin inhibitory capacity, and Pi typability. The alpha1-antrypsin level and trypsin inhibitory capacity were not significantly correlated with morgue time, serum storage time, or pH, and mean values were within the expected ranges. A significant decrease in Pi typability occurred when pH was less than 7.0. Moreover, while most (86%) of the sera stored for 2 to 2 1/2 years were typable, only 30% of those stored for more than four years were typable. Determination of alpha1-antitrypsin and trypsin inhibitory capacity are possible with the use of stored postmortem blood. Pi typing is usually possible, provided sera are not acidic and are examined within 2 1/2 years.


Analytical goals in clinical chemistry: their relationship to medical care. Proceedings of the Subcommittee on Analytical Goals in Clinical Chemistry. World Association of Societies of Pathology, Ciba Foundation, London, England (U. K.), April 25-28, 1978. The discipline of clinical chemistry is undergoing dramatic changes. The force for change stems from a worldwide recognition of the importance of chemical analyses in medical practice. In the past, developments in clinical chemistry were initiated from a technical perspective. This report provides an alternative approach, starting from the patient and then defining the role of the clinical chemistry laboratory in patient care. The discussions attempt to define the relationship between patient care and analytic goal development in statistical terms. This approach provides a method for appraising the needs of medical care in quantitative terms from the perspective of the physician and the patient. The approach also provides guidance for identifying areas where further technical advance is required, as well as areas where present laboratory performance is adequate for patient care in terms of analytic quality.


Comparison of hemoglobins Wood (alpha 2 beta 2 97 leu) and Malmö (alpha 2 beta 2 97 gln). Diagnostic value of citrate agar electrophoresis. Diagnostic value of citrate agar electrophoresis. Am J Clin Pathol 71:668-671, 1979. Of approximately three dozen hemoglobin variants that have greater than usual oxygen affinity, nearly half are inseparable from hemoglobin A by electrophoresis at pH 8.6. A comparison of hemoglobins Wood (alpha2beta297leu) and Malmö (alpha2beta297gln) is of interest from several standpoints. They represent similar substitutions at the identical locus in the beta chain. They result in identical clinical and hematologic manifestations. Oxygen affinities of these variants are identical. Both are poorly resolved from hemoglobin A by electrophoresis at pH 8.6. The position of each is identical when studied by isoelectric focusing in polyacrylamide gel. Finally, they are easily distinguished by citrate agar electrophoresis at pH 6.2. The excellent resolution of hemoglobins Malmö and Wood from each other results neither from difference in charge, nor size, nor in quaternary structure. This technic provides a simple but effective means for identifying and differentiating these hemoglobin variants. Comparison with the results of citrate agar electrophoresis of other high oxygen-affinity hemoglobins indicates that the findings for hemoglobins Malmö and Wood are unique and unambiguous.


Characterization of a virus isolated from a case of human infectious hepatitis. A new viral agent, isolated from the serum of an infectious hepatitis patient and designated as Agent II-B, was extensively studied in in vitro and in vivo systems. Agent II-B multiplied well in primary and serial animal cell cultures and in embryonated hen's eggs. Quantal and quantitative infectivity assays were performed in monolayers of African green monkey kidney cells. Effective concentrations of 5-iodo-2-deoxyuridine and guanidine hydrochloride did not inhibit the multiplication of Agent II-B, although 2-hydroxybenzyl-benzimidazole was an effective inhibitor. Essential lipids were not detected. The diameter of the agent is 16-25 nm and its buoyant density in CsCl equilibrium density gradients was 1.35 gm/ml. Neutralization test results did not reveal antigenic relatedness between Agent II-B and known human picornaviruses. Apparently, this new viral agent is a picornavirus which possesses the capacity to multiply in unexpectedly diverse cell types.


Control factors of granulopoiesis in human serum. The role of serum factors in the modulation of production of colony-stimulating activity (CSA) has been investigated. A factor has been described, and partially characterized, in human serum that has the capacity to stimulate increased synthesis and release of CSA by human mononuclear cells (MNC). MNC RNA and protein synthesis are required to demonstrate this effect of serum, but DNA synthesis and mitotic division are not required. The factor in serum resulting in this effect is a heat-labile protein with a molecular weight slightly greater than that of CSA.


Heritable fragile sites on human chromosomes I. Factors affecting expression in lymphocyte culture. The expression of heritable fragile sites on human chromosomes has been shown to be dependent upon composition of the tissue medium for sites at 2q11, 10q23, 11q13, 16p124, 20p11 and Xq27 or 28 but not for the site at 16q22. Expression of the fragile sites is inhibited by folic acid, thymidine, folinic acid, and probably bromodeoxyuridine, and induced by methrotrexate. In addition, there is a correlation between frequency of expression of the sites and pH of the culture medium for the sites on 2q, 10q and Xq. Possible reasons for these findings are discussed, and a definition and classification of fragile sites is proposed.


Effect of pH on the stability of heparin in 5% dextrose solutions. The effect of pH and time on the stability of heparin sodium in dextrose 5% in water (D5W) injection and in dextrose 5% in 0.45% sodium chloride injection was studied. Admixtures of heparin sodium 5,000 units/250 ml were tested after 0, 10, 20 and 30 minutes and 1, 2, 6, 12 and 24 hours of storage at room temperature. The pH of the carrier solutions was adjusted to 2, 4 or 9 prior to adding the heparin sodium. Heparin activity was measured using a thrombin clotting time assay. Samples were tested for pH changes at the same times. No substantial changes in heparin activity over the 24-hour period occurred with any of the pH-adjusted solutions. The pH of the heparin-D5W admixtures remained constant over time. The two carrier solutions, over a pH range of 2 to 9, appear to be suitable vehicles for heparin sodium.


Phenothiazine analgesia--fact or fantasy? Double-blind clinical trials involving the use of phenothiazines as analgesics or potentiators of analgesics (aspirin, meperidine, morphine sulfate) and adverse effects of phenothiazines are reviewed and evaluated. Promethazine, promazine and propiomazine were not found to possess analgesic or potentiating properties. One chlorpromazine study contained important design and reporting deficiencies which precluded a recommendation for use of chlorpromazine in the treatment of pain. Methotrimeprazine was determined by numerous authors to have analgesic properties; however, most of the studies also were deficient in design or data presented, or both. Adverse reactions to phenothiazines, including hypotension, sedation, drowsiness, extrapyramidal symptoms, tardive dyskinesia, cardiac toxicity and agranulocytosis, are often more common and severe than those attributed to narcotic analgesics. Because of the lack of data supportive of analgesic activity and the adverse reactions associated with phenothiazines, use of these agents in the management of pain should be discouraged. The prophylactic use of phenothiazine for narcotic analgesic-induced emesis also is, in most cases, a questionable practice.


Efficacy of antiparkinson agents in preventing antipsychotic-induced extrapyramidal symptoms. The types of extrapyramidal syndrome (EPS) reactions produced by antipsychotic agents and the prophylactic use of antiparkinson agents in preventing EPS are reviewed. EPS are classified as akathisias, dystonias, parkinson-like symptoms and tardive dyskinesia, and have a varied incidence reported to range from 10.6 to 100%. Incidence may vary with age, gender, drug and dosage. The prophylactic use of antiparkinson agents to prevent EPS is controversial. Many psychiatrists believe the effect of EPS on patients is more harmful than the side effects of anticholinergics, whereas others believe that because of side effects, increased cost, greater risk to tardive dyskinesia and improper use, the use of antiparkinson agents cannot be justified. Most studies of prophylactic use of antiparkinson agents have lacked adequate control groups, adequate blinding procedures for investigators rating EPS, uniform definitions of EPS, random sampling and careful reporting of group characteristics such as dosage and drugs received. There is a lack of definitive studies of the value of antiparkinson agents in preventing the occurrence of EPS in patients receiving antipsychotics. A large, multicenter study should be undertaken to resolve the issue.


In vitro acid reactivity of three commercial antacid tablets. The in vitro acid reactivity of three commercial brands of aluminum and magnesium hydroxide antacid tablets was determined by two methods. A pH-stat test was used to examine the rate and extent of acid neutralization at a constant pH of 3.0. A modified Rossett-Rice test was used to record the length of time during which the antacid products maintained the pH of a simulated gastric solution at between 3.0 and 5.0. Acid neutralization by product A was faster and more complete than that by product B or C. The percent of theoretical acid consuming capacity at 30 minutes of product A (86.8%) was significantly greater than that of product B (56.1%) and product C (57.0%) tablets. The 32-minute Rosett-Rice time A was significantly longer than the 16- and 12-minute times of products B and C, respectively. The differences observed may be attributed to different reactivities of the raw materials used in the products, or formulation and processing variables. It is not known how the data relate to in vivo performance.


Reduced uterine blood flow and fetal hypoxemia with acute maternal stress: experimental observation in the pregnant baboon. The effects of maternal hyperexcitability on the fetus were studied in 17 baboons. In the period of agitation, induced by stressful stimulus such as exposure to bright light or by clamping of the toe, the mother exhibited an increase in arterial blood pressure and, in some instances, arrhythmia. These changes were accompanied by an increased uterine activity and reduced uterine blood flow, and resulted in a decrease in heart rate and arterial oxygenation in all fetuses. Fetal recovery was prompt after maternal agitation was terminated, either by removal of the stimulus or by sedation with pentobarbital or nitrous oxide. This sedation also prevented a decrease in uterine blood flow when stress was repeated.


Fetal breathing: a review.

Fetal breathing: a review. The history, physiology, pharmacology, and clinical application of fetal breathing are reviewed. Early optimism that measurements of fetal breathing would aid in perinatal management decisions has remained unfulfilled. Problems to be overcome include standardization of technique and measurement parameters and allowance for physiologic variations. Recent research developments indicate that the likely role of fetal breathing in perinatal medicine will be in conjunction with other established tests of fetal health.


Beta-agonists and secretory cell number and intracellular glycoproteins in airway epithelium. The effect of isoproterenol and salbutamol. This study describes the effect of systemic administration of the beta-adrenergic agonists isoproterenol and salbutamol on the secretory cell populations in seven regions of rat airway epithelium (three extrapulmonary and four intrapulmonary) and on the size of salivary glands and heart. Isoproterenol (a nonselective beta-adrenergic agonist) significantly increases secretory cell number in all airway regions except the midtrachea; salbutamol (a selective beta 2 agonist) increases secretory cell number only in proximal and peripheral regions. The absolute number of secretory cells is greatest in the most peripheral region after isoproterenol administration and in the most proximal region after salbutamol, although both drugs produce the greatest relative increase at the periphery. In proximal and, particularly, peripheral regions, the increase by isoproterenol (less than 3- and 14-fold, respectively) is greater than by salbutamol (less than 2- and less than 3-fold, respectively). In all airway regions, both drugs modify intracellular glycoprotein in the secretory cell population; within a given region, modification is much the same. In the most proximal region, the population of cells synthesizing only granules of neutral glycoprotein significantly increases while in other regions increase is in cells synthesizing only granules of acid. A significant shift in glycoprotein synthesis occurs whether or not the secretory cell population is increased, which suggests that existing as well as newly appearing cells modify their product. Isoproterenol significantly increases the size of the parotid and submaxillary glands; salbutamol increases the size of the parotid only. Isoproterenol significantly increases the weight of both ventricles of the heart; salbutamol has no such effect.


The ultrastructure of the human epidermis in chronic graft-versus-host disease. The epidermal ultrastructure of 11 allogeneic bone marrow recipients with chronic graft-versus-host disease (GVHD) was compared with that of 4 recipients without chronic GVHD. This electron microscope study revealed three patterns of epidermal injury typical of chronic GVHD. The first type was a nonacantholytic (nondissecting) injury with a prominent cellular infiltrate consisting primarily of lymphocytes accompanied by a few macrophages. The second type was an acantholytic (dissecting) injury with a prominent infiltrate, while the third was a nondissecting injury with a sparse infiltrate. Broad-zone contact was observed between lymphocytes and all epidermal cell types as well as between other lymphocytes and macrophages. Point contact was only observed between lymphocytes and epidermal cells. Lymphocytes appeared to detach desmosomes from adjacent keratinocytes by isolating them with cytoplasmic projections, a phenomenon not previously described. Typical damage to the epidermal cells in the basal and spinous layers consisted of either swelling of the organelles or condensation of the cytoplasm and nucleus. In the keratinocyte, the condensation reaction resulted in the formation of colloid bodies, some of which were phagocytized by macrophages. Besides the cytolytic events, a concurrent stimulatory reaction occurred in the epidermal cells. The number of melanosomes in melanocytes and of Langerhans cell granules and dense bodies in the Langerhans cells all increased. Extensive areas of replication and disruption of the basal lamina were subjacent to areas of necrosis in the basal layer.


Clinical implications of state-dependent learning. Researchers have found that state-dependent learning is associated with the administration of a wide variety of drugs. Recent data suggest that similar phenomena may occur secondary to endogenous changes in neuroregulatory substances. The authors point out that awareness of such changes in cognitive processing strategies and abilities should help to further our understanding of the phenomenology of psychiatric states and should generate psychotherapeutic techniques designed to maximize the transfer of information across psychiatric states.


The form and content of schizophrenic hallucinations. To evaluate changes in the hallucinatory experience, the author analyzed the form and content of the hallucinations of 10 schizophrenic patients during two phases of their illness, the acute phase and remission. In both test periods the patients experienced auditory hallucinations as objectively real and as directing their thoughts and behavior. Nonauditory hallucinations appeared more common than cursory examination would suggest. The patients were able to stop auditory hallucinations during remission but not during the acute phase of their illness. Hallucinatory content was threatening and isolating during the acute phase but supportive and socially focused during remissions.


Informed consent for neuroleptic therapy. The author suggests that the frequency and severity of tardive dyskinesia in patients treated with neuroleptic drugs requires that informed consent be obtained from all patients receiving such treatment. The three basic conditions for obtaining informed consent are reviewed with discussion of some of the ethical problems encountered in the informed consent procedure. Most of these problems will be resolved if specific, written, informed consent is obtained from the patient, or his representative, within six weeks of initiating therapy, although in some cases questions may be raised about the very possibility of obtaining consent.


Relation between transmucosal potential difference, ionic flux, and the intraluminal supply of H+ in the ferret stomach. To investigate the relation between gastric transmucosal potential difference, ionic flux, and the intraluminal concentration of H+, test solutions containing various concentrations of HCl were instilled into ferret stomachs both before and after exposure to various doses of acetic acid. The changes in ionic composition of the test solution were determined and the transmucosal potential difference was recorded throughout each experiment. The results showed that after exposure to the organic acid (1) ionic flux was proportional to the concentration of acetic acid used and was a direct function of the concentration of H+ within the lumen, and (2) the decrease in transmucosal potential difference was proportional to the concentration of acetic acid used but did not appear to be related to the subsequent magnitude of ionic flux. It is suggested that the decrease in gastric transmucosal potential difference that follows exposure of the gastric mucosa to an organic acid is a measure of the increase in mucosal permeability, whereas the subsequent ionic movements are governed by the concentration of H+ within the gastric lumen.


Post-partum sterilisation--an anaesthetic hazard? A case of acid aspiration pneumonitis which occurred in the puerperium is described. Gastric aspirate was collected in fifty patients undergoing postpartum sterilisation. Results of pH and volume suggest these patients should be regarded as potential candidates at risk of developing Mendelson's syndrome.


Postoperative analgesia.

Postoperative analgesia. Postoperative analgesia both by drugs and regional techniques is reviewed. In the United Kingdom in the last 25 years or more there has been little advance on either front. Some marginal improvement in regard to drugs might be brought about by better education of both doctors and nurses and better patient contact. Extradural analgesia and intercostal block do not offer a complete solution, though a judicious increase in the use certainly of the former might be beneficial. The problem awaits a radical new approach.


Pre-anesthetic cimetidine alteration of gastric fluid volume and pH. Gastric volume and pH were studied immediately after induction of anesthesia and endotracheal intubation in 101 elective surgical patients. Of 44 patients not given cimetidine, 82% had a gastric pH less than 2.5 with a mean pH of 1.6; 45% of these patients had a gastric aspirate pH less than 2.5 associated with a volume exceeding 25 ml. In 57 patients premedicated with intravenous cimetidine at variable intervals (15 to 60 minutes) prior to induction of anesthesia, a significant time-dependent increase was noted in gastric pH (p less than 0.001) together with a decline in gastric volume (p less than 0.001). Of the patients given intravenous cimetidine (mean 4.5 mg/kg) 45 minutes prior to induction of anesthesia, 90% had a gastric pH greater than 2.5. The increase in gastric pH after cimetidine administration would result in a reduced chemical pulmonary reaction should aspiration occur during induction of anesthesia.


Dopamine treatment of spinal hypotension decreases uterine blood flow in the pregnant ewe. In seven pregnant ewes, 3--5 min of hypotension resulting from spinal anesthesia decreased uterine blood flow 17 per cent. Dopamine, in doses sufficient to maintain blood pressure at control values (20--40 micrgram/kg/min) for 3--5 min, further decreased uterine blood flow to 56 per cent less than control and increased uterine vascular resistance to 50 per cent more than control. Following 30 min of hypotension, 30 min of dopamine administration in doses sufficient to restore blood pressure to control values (5--20 microgram/kg/min) similarly decreased uterine blood flow to 29 per cent less than control and increased uterine vascular resistance to 35 per cent more than control.


Effects of changes in maternal--fetal pH on the transplacental equilibrium of bupivacaine. Increases in the maternal-fetal pH gradient that may occur during labor and delivery may increase the fetal concentration of local anesthetics. The authors evaluated effects of pH changes on the transplacental concentration equilibrium of bupivacaine. They increased the maternal-fetal pH gradient in each of six pregnant ewes from a control value of 0.15 to 0.54 by hyperventilating the lungs of the ewe and infusing lactic acid into her fetus. After infusion of bupivacaine, 0.15 mg/kg, intravenously into the mother, the drug rapidly appeared in fetal blood, with values significantly increased over control values at 1 and 5 min. The fetal/maternal (f/m) ratios were increased significantly at 5, 15, and 30 min. The f/m ratios had stabilized by 15 min in both control and experimental states, suggesting that equilibrium had been achieved. The consistently low f/m ratios are explained by the presumed similarity of the ovine maternal and fetal protein binding rates to those of man. It is concluded that the maternal and fetal pH values are major factors in the determination of the f/m ratios.


[Effect of carbon and nitrogen source additions on the development of a productive strain of P. nigricans Thom and on the level of adenylates in its mycelium]. Growth of the cultured strain og P. nigricans and dynamics of the adenylate levels in its mycelium on mineral media with 2 per cent of glucose were studied in relation to the means and time of addition of glucose, NaNO3 or their mixture to the medium. It was shown that the maximum yield of the mycelium could be obtained with addition of glucose once at the moment of inoculation. The mixture of glucose with NaNO3 provided even higher yields of the biomass but only with its fractional addition. Introduction of additional amounts of NaNO3 at the moment of inoculation and during the growth phase (5 days) inhibited the subsequent development of the culture providing stable levels of ATP and ADP, while introduction of NaNO3 on the 7th day stimulated the culture growth and the antibiotic yield. The use of NaNO3 in the mixture with glucose eliminated inhibition and increased the ratio of ATP to ADP and the antibiotic yield.


[Aminoglycoside-3'-phosphotransferase from Actinomyces fradiae. Its isolation, purification and properties]. Aminoglycoside phosphotransferase was isolated from the mycelium of Act. fradiae, the neomycin-producing organism, with paromomycin, neomycin and to a less extent ribostamycin being substrates of aminoglycoside-phosphotransferase. It was purified to homogenous state. The maximum activity of the enzyme preparations was observed at pH 7.7--7.8;KM for neomycin and paromomycin was about 20 micron and KM for ATP was 150 micron. Mg2+ ions were necessary for the enzyme activity. None of the divalent cations tested could replace the magnesium ions in the reaction of phosphorylation catalyzed by the enzyme. High sensitivity to the ionic strength of the buffer was characteristic of the enzyme. It lost about 80 per cent of the initial activity at a concentration of KC1 equal to 1.0 M. The molecular mass of the enzyme from the mycelium of Act. fradiae was determined by the method of gel-filtration through sefadex G-100. It was about 22,000. High stability was characteristic of the enzyme. The fingings indicate that aminoglycoside phosphotransferase from Act. fradiae differs from the described aminoglycoside-3'-phosphotransferases isolated from antibiotic resistant bacteria.


[Aminoglycoside-3'-phosphotransferase from Actinomyces fradiae. The identification of its inactivation product]. Neomycin phosphate was obtained as a result of neomycin phosphorylation with aminoglycoside-phosphotransferase from Act. fradiae. It was isolated from the reaction mixture and purified. Successive ion exchange chromatography on columns with Amberlite IRC-50 (NH+4 form), Dowex 1 X 10 (OH- form) and Amberlite CG-50 (NH+4 form) was used for purification of the inactivation product. The findings of the elementary analysis of neomycin phosphate showed the presence of 1 mole of phosphorus per 1 mole of the antibiotic. From the results of the chemical analysis, IR- and NMR-spectrometry neomycin phosphate and neamine phosphate obtained from it by methanolysis were identified as neomycin-3'-phosphate and neamine-3'-phosphate, respectively. The data indicate that the enzyme isolated from Act. fradiae is aminoglycoside-3'-phosphotransferase.


Properties of beta-glucan synthetase from Saccharomyces cerevisiae. Properties of beta-glucan synthetase from S. cerevisiae were studied. The enzyme exhibited optimal activity at pH 6.7 and 24 C. Km for UDP-glucose was 0.12 mM. Addition of Mg++ or Mn++ stimulated its activity by 60% and 21% respectively. High concentrations of EDTA and hydroxyquinoline were inhibitory. Glucan synthetase was fully active in cell-free extracts. Small concentrations of trypsin or subtilopeptidase A from Bacillus subtilis, caused only a slight increase in glucosyl transferase activity, but larger concentrations destroyed beta-glucan synthetase. Acid proteases were neither stimulatory nor destructive. Thus it seems unlikely that beta-glucan synthetase exists in a zymogen form. Glucan synthetase was unstable. It was inactivated more rapidly at 28 C than at 0 C. The presence of substrate, beta-glucan or the protease inhibitors PMSF, Antipain or Pepstatin A did not protect beta-glucan synthetase from inactivation. Glucan synthetase was not stimulated by addition of cellobiose or beta-glucans. The synthesis of beta-glucans was competitively inhibited by UDP (Ki = 0.45 mM). Glucono-delta-lactone, a known inhibitor of beta-glucosidases was a strong non-competitive inhibitor of beta-glucan synthetase.


Nitroblue tetrazolium-dye reduction by rat peritoneal macrophages during the uptake of Diplococcus pneumoniae, type VI. The relationship between the rate of particle ingestion and the rate of Nitroblue Tetrazolium (NBT)-dye reduction by macrophages was studied after incubation of peritoneal exudate macrophages with heat-killed type VI pneumococci. The adherence to a polyethylene surface of the macrophages during the uptake of the pneumococci was determined as well. In some experiments pneumococci opsonized with heat-stable opsonins were used as material to be ingested. The NBT-dye reduction and the surface adherence of the macrophages was enhanced when ingesting normal heat-killed pneumococci. During the uptake of opsonized, heat-killed pneumococci the macrophages showed an unaltered NBT-dye reduction and less adherence to a polyethylene surface as compared with macrophages incubated with normal heat-killed pneumococci. This implies that using opsonized pneumococci the quantitative NBT-dye reduction assay is not reliable as a parameter for macrophage phagocytosis, because the uptake was in fact enhanced. The surface adherence of the macrophages did not reflect the enhanced ingestion of opsonized bacteria either.


Urease-producing species of intestinal anaerobes and their activities. Urease activities of anaerobic bacteria that constituted predominant gut flora were examined. It was demonstrated that some strains of Eubacterium aerofaciens, E. lentum, and Peptostreptococcus products produced urease. They were the most numerous species in human feces. All strains of Bifidobacterium infantis and some strains of Bacteroides multiacidus, B. bifidum, Clostridium symbiosum, Fusobacterium necrophorum, F. varium, Lactobacillus fermentum, Peptococcus asaccharolyticus, and P. prevotii produced urease. The optimum pH of the Lactobacillus urease was found to be 4.0, whereas the pH value of B. multiacidus urease was 8.0.


Emulsifier of Arthrobacter RAG-1: isolation and emulsifying properties. The oil-degrading Arthrobacter sp. RAG-1 produced an extracellular nondialyzable emulsifying agent when grown on hexadecane, ethanol, or acetate medium. The emulsifier was prepared by two procedures: (i) heptane extraction of the cell-free culture medium and (ii) precipitation with ammonium sulfate. A convenient assay was developed for measurement of emulsifier concentrations between 3 and 75 micrograms/ml. The rate of emulsion fromation was proportional to both hydrocarbon and emulsifier concentrations. Above pH 6, activity was dependent upon divalent cations; half-maximum activity was obtained in the presence of 1.5 mM Mg2+. With a ratio of gas oil to emulsifier of 50, stable emulsions were formed with average droplet sizes of less than 1 micron. Emulsifier production was parallel to growth on either hydrocarbon or nonhydrocarbon substrates during the exponential phase; however, production continued after growth ceased.


Bacteriocin production by Clostridium acetobutylicum in an industrial fermentation process. High titers of a noninducible bacteriocin were produced by Clostridium acetobutylicum in a molasses fermentation medium used for the industrial production of solvents. Release of the bacteriocin towards the end of the exponential growth phase was accompanied by lysis of the culture and inhibition of the production of solvents. The producer cells were sensitive to the bacteriocin, which only affected other C. acetobutylicum strains and a Clostridium felsineum strain. The thermolabile bacteriocin was not inactivated by protease enzymes and had no optimum stability between pH 4 and 5. The sedimentation coefficient of the bacteriocin was 6S.


Heat resistance of Byssochlamys ascospores. Ascospores from 25 strains of Byssochlamys were studied for their ability to resist heat treatment in a standard defined medium. Seven of these were able to survive heating at 90 degrees C for 25 min or longer, when initial numbers were frequently near 10(6)/ml. Ascospores from five resistant strains suspended in the medium at pH 5.0 were usually more resistant than those at pH 3.6. Rapid heat inactivation occurred for one strain at pH 6.6. Nonlogarithmic heat death rate was observed in all strains tested.


Clostridium botulinum growth and toxin production in tomato juice containing Aspergillus gracilis. The ability of spores of one type A and one type B strain of Clostridium botulinum to grow and produce toxin in tomato juice was investigated. The type A strain grew at pH 4.9, but not at pH 4.8; the type B strain grew at pH 5.1, but not at pH 5.0. Aspergillus gracilis was inoculated along with C. botulinum spores into pH 4.2 tomato juice; in a nonhermetic unit, a pH gradient developed under the mycelial mat, resulting in C. botulinum growth and toxin production. In a hermetic unit, mold growth was reduced, and no pH gradient was detected; however, C. botulinum growth and low levels of toxin production (less than 10 50% lethal doses per ml) still occurred and were associated with the mycelial mat. The results of tests to find filterable or dialyzable growth factors were negative. It was demonstrated that for toxin production C. botulinum and the mold had to occupy the same environment.