Intermediate sucrose octa-acetate sensitivity suggests a third allele at mouse bitter taste locus Soa and Soa-Rua identity

Mice have been characterized as either tasters or non-tastersof the bitter compound sucrose octa-acetate(SOA). However, 11of 17 supposedly non-taster inbred strains were found to avoid1 mM SOA. All 17 strains were indifferent to 0.1 mM SOA. Tasterstrains avoided both concentrations. The intermediate phenotypewas dubbed demitaster. A consistent phenotypic dominance orderwas found in crosses among both inbred and outbred strains (taster> non-taster > demitaster). Demitasters were found (withtasters) in an outbred strain showing monogenic segregationfor SOA avoidance. This, plus monogenic segregation in a back-crossof taster to demitaster inbred strains, suggested a third alleleat the Soa locus (Soa^c). Demitaster allelism was supported bythe strong associations found in 15 strains between the threeSOA phenotypes and HindIII restriction fragment patterns forthe closely linked Prp (proline rich protein) loci. SOA demitasterstrains were also intermediate in raffinose undeca-acetate (RUA)avoidance. Furthermore, B6.SW-Soa² congenic mice avoided notonly SOA, but RUA and eight other acetylated sugars. A previouslyproposed separate RUA-sensitivity gene (Rua) thus appeared tobe redundant.     

Metabolic regulation in the facultative methylotroph Arthrobacter P1. Growth on primary amines as carbon and energy sources

During growth of the facultative methylotroph Arthrobacter P1 on methylamine or ethylamine both substrates are metabolized initially in an identical fashion, via the respective aldehydes. The regulatory mechanisms governing the synthesis and activities of enzymes involved in amine and aldehyde utilization were studied in substrate transition experiments. Transfer of ethylamine-grown cells into a medium with methylamine resulted in immediate exeretion of low levels of formaldehyde (max. 0.5 mM) and formate. In the reverse experiment, transfer of methylaminegrown cells into a medium with ethylamine, excretion of much higher levels of acetaldehyde (max. 3.5 mM) occurred. These different levels of aldehyde accumulation were also observed in studies with mutants of Arthrobacter P1 blocked in the synthesis of hexulose phosphate synthase or acetaldehyde dehydrogenase. In wild type Arthrobacter P1, aldehyde production resulted in rapid induction of the synthesis of enzymes involved in their degradation but also in temporary inhibition of further amine utilization and growth. The latter aetivities only resumed at normal rates after the disappearance of the aldehydes from the cultures. Acetaldehyde utilization resulted in intermittent excretion of ethanol and acetate, whereas formaldehyde utilization resulted in further accumulation of formate.During growth of Arthrobacter P1 in the presence of methylamine accumulation of toxic levels of formaldehyde is prevented because of the rapid synthesis of hexulose phosphate synthase to high activities and, in transient state situations, by feedback inhibition of formaldehyde on the activities of the methylamine transport system and amine oxidase.Abbreviations DTNB 5,5-dithiobis-(2-nitrobenzoate) - HPS hexulosephosphate synthase - MS mineral salts - RuMP ribulose monophosphate     

A spectrophotometric method for the determination of the kinetic parameters of NH+4 uptake by yeast cells

Abstract The kinetic parameters of NH⁺₄-uptake in yeast cells were determined by a method that is based on the following changes in the external NH⁺₄ concentration in cell suspensions by using NADH-dependent glutamate formation from NH⁺₄ and 2-oxoglutarate. The kinetics of the observed NADH oxidation were analyzed by computer and enabled an estimation of V _max and K _m of the NH⁺₄-uptake system of the cells.     

Biosynthetic pathways of Vibrio succinogenes growing with fumarate as terminal electron acceptor and sole carbon source

1. With fumarate as the terminal electron acceptor and either H₂ or formate as donor, Vibrio succinogenes could grow anaerobically in a mineral medium using fumarate as the sole carbon source. Both the growth rate and the cell yield were increased when glutamate was also present in the medium.
2. Glutamate was incorporated only into the amino acids of the glutamate family (glutamate, glutamine, proline and arginine) of the protein. The residual cell constituents were synthesized from fumarate.
3. Pyruvate and phosphoenolpyruvate, as the central intermediates of most of the cell constituents, were formed through the action of malic enzyme and phosphoenolpyruvate synthetase. Fructose-1,6-bisphosphate aldolase was present in the bacterium suggesting that this enzyme is involved in carbohydrate synthesis.
4. In the absence of added glutamate the amino acids of the glutamate family were synthesized from fumarate via citrate. The enzymes involved in glutamate synthesis were present.
5. During growth in the presence of glutamate, net reducing equivalents were needed for cell synthesis. Glutamate and not H₂ or formate was used as the source of these reducing equivalents. For this purpose part of the glutamate was oxidized to yield succinate and CO₂.
6. The α-ketoglutarate dehydrogenase involved in this reaction was found to use ferredoxin as the electron acceptor. The ferredoxin of the bacterium was reoxidized by means of a NADP-ferredoxin oxidoreductase. Enzymes catalyzing the reduction of NAD, NADP or ferredoxin by H₂ or formate were not detected in the bacterium.

     

Der Serotoninstoffwechsel in einzelnen Hirnteilen vom Tupaia (Tupaia belangeri) bei soziopsychischem Stress

Zusammenfassung Bei sozialem Stress nehmen die Konzentrationen von Serotonin (5-HT) und 5-Hydroxyindolessigsäure (5-HIAA) in den einzelnen Hirnteilen von Tupaias zu. Diese Zunahme läßt sich durch eine erhöhte Stoffwechselgeschwindigkeit (turnover) erklären.Die Änderungen in der Stoffwechselgeschwindigkeit der einzelnen Hirnteile hängen von der Art der sozialen Belastung ab.Schon Kampf mit Unterwerfung (zweimal täglich) führt zu einem schnelleren Stoffwechsel des 5-HT in allen Hirnteilen. Belastet man die Tiere außer durch Kampf noch dadurch, daß sie das Tier, das sie besiegt hat, zwischen den Kampfpausen sehen können, so wird der Stoffwechsel des 5-HT in einzelnen Hirnteilen (Septum, Frontaler Cortex) nochmals erhöht.Eine mögliche funktioneile Bedeutung dieser Veränderungen wurde diskutiert.

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Serotonin metabolism in discrete brain areas and sociopsychic stress in tupaia (Tupaia belangeri)

Summary The physiological state of the animals depends not only on ecological factors. It is also determined by the social interactions of the animals. Some of these social interactions are harmful to the physiological state of the individuals. The aim of this work was to separate the physical loads of social interactions from the mere psychic ones and to correlate them with neurochemical changes. For this purpose the concentration levels of serotonin (5-HT) and 5-hydroxyindoleacetic acid (5-HIAA) were measured in discrete brain areas.To separate the psychic from the physical loads male tupaias (Tupaia belangeri) were exposed two times a day (at 9.30 and 13.30 h) for two minutes to the attacks of an experienced fighter. The animals were defeated during these attacks. One group of the defeated animals was in optical contact with the experienced fighter throughout the day (12 hours; Stresstiere). The other group of the defeated animals was optically separated from the experienced fighter after each fight (Kampftiere).After four days the animals were decapitated at 15.00 h, 5.5 hours after the last fight (on the fourth day the animals were not allowed to fight at 13.30 h). Animals which had not fought were decapitated at the same time and served as controls. 5-HT and 5-HIAA levels were estimated within two days after decapitation.During the test the animals lose body weight. Animals who are only exposed to attacks (Kampftiere) loose 2.8±2.8%; animals who are exposed to attacks and optical contact with the experienced fighter (Stresstiere) lose 11.8±3.4%.The concentration levels of 5-HT and 5-HIAA in both groups are elevated compared with controls. This fact indicates a higher turnover of these substances. The concentration levels are most elevated in parts of the brain of defeated animals exposed to continuous optical contact with the experienced fighter (Stresstiere). These differences are most prominent in the septum and the frontal cortex (s. p. 60).A model for the physiological meaning of these changes is discussed. It was assumed that the activity of the brain is controlled by a system which is functionally analogous to the peripheral autonomous nervous system. 5-HT and norepinephrine should act on antagonistic branches of such a system in central nervous system. Under stress conditions the activity of both branches should be enhanced.

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Herrn Prof. Dr. H. Autrum möchte ich für die Überlassung des Themas, für sein stetes Interesse und anregende Diskussionen danken. Die Untersuchungen wurden durch Sachbeihilfen gefördert, die die Deutsche Forschungsgemeinschaft Herrn Prof. Dr. H. Autrum zur Verfügung stellte.

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Der Firma Sharp & Dohme GmbH danke ich für die kostenlose Überlassung von 15 g Probenecid.     

Influences on the density and dispersion of bumble bee nests (Hymenoptera: Apidae)

To determine some of the influences on nesting densities of bumble bees I examined the dispersion and occupancy of 35 nests in an old field on Amherst Island, Ontario, Canada and 39 nests found by Cumber (1953) in England. Densities up to 1 nest per 200 m² were observed. Overall, Cumber's nests were randomly dispersed, but nests on the ground surface were aggregated. Nests of surface-nesting species were consequently more aggregated than expected at random, whereas surface-nesting and underground-nesting species seldom nested near one another. This distribution of nests probably reflects the nesting habits of the rodents that originally constructed the nests, rather than interactions between bees. On Amherst Island, abnormally wet spring weather apparently hampered normal colony development, particularly of underground-nesting bees based on a low frequency of underground nests, scarcity of workers of two underground-nesting species compared with the relative abundance of spring queens, and delayed appearance of workers of all species relative to the previous year. The nests of two common species were randomly dispersed, but those of a third species were regularly distributed, suggesting that intraspecific interactions may limit nesting density for this species.     

Hypoxic induction of Ca2+-dependent action potentials in small pulmonary arteries of the cat

Small pulmonary arteries (less than 300 micron) from cats were mounted in myographs to record mechanical and electrical responses to hypoxia. When these preparations were exposed to a PO2 of 30-50 Torr after equilibration at 300 Torr they consistently developed active force, which increased or decreased in amplitude as [Ca2+] was raised or lowered, respectively, and was blocked on addition of verapamil. Intracellular electrical recording with glass microelectrodes demonstrated membrane depolarization and action potential generation when PO2 was lowered. Steady-state voltage vs. applied current curves obtained before and during hypoxia showed a significant reduction in input resistance. The relationship between membrane potential and extracellular K+ was not different during hypoxia compared with control, suggesting that there were not marked changes in K+ permeability under this condition. In the presence of verapamil to block Ca2+ inward current the hypoxia-induced action potentials were abolished concomitant with partial membrane repolarization. The results of these studies suggest that in certain isolated pulmonary arteries hypoxia induces contraction by a mechanism involving an increased Ca2+ conductance. These data suggest that the sensor involved in hypoxic pulmonary vasoconstriction may lie within the vessel wall and somehow mediates changes in smooth muscle ionic conductances.     

On-line monitoring of enzymes in downstream processing by flow injection analysis (FIA)

Flow injection analysis (FIA) has been employed to automate enzyme assays for formate dehydrogenase (FDH) and l-leucine dehydrogenase (l-LeuDH). Coupled to a special sampling device the FIA assays were used to monitor on-line downstream processes, e.g. disintegration of microbial cells and cross-flow filtration of cell homogenates.