Yeast mutants resistant to ethidium bromide

Summary Yeast mutants resistant to ethidium bromide have been isolated among sensitive grande cells (⁺) for their ability to grow on glycerol in the presence of the dye. Mutant cells are also resistant to acriflavin and do not yield petites (^-) when grown on galactose with the mutagen. Genetic analysis reveals that resistance to ethidium bromide is controlled by a cytoplasmic factor, carried by, or linked to, the determinant (mitochondrial DNA). The expression of resistance to ethidium bromide seems to be related to the presence in the cell of a product of mitochondrial protein synthesis. It is concluded that some mitochondrial DNA sequence is involved in the resistance to ethidium bromide of yeast mitochondria.     

Increase of plasma membrane oxidoreductase activity is not correlated with the production of extracellular Superoxide radicals in human Namalwa cells

Summary We have considered the regulatory interrelationship of the plasma membrane oxidoreductase (PMOR) system and the mitochondrial respiratory capacity of human Namalwa (lymphoblastoid) cells. To this end, we made use of mitchondrially respiratory competent (⁺) cells and ⁰ cells, which lack mitochondrial DNA (mtDNA) and consequently mitochondrial respiratory activity. NADH-fer-ricyanide reductase activity of the PMOR system is increased 3-fold in ⁰ Namalwa cells compared to ⁺ cells. It is also shown for the first time that addition of coenzyme Q₁₀ and coenzyme Q₁₀-ana-logues, which can rescue ⁰ Namalwa cells in the absence of pyravate, gives rise to a further 2–3-fold increase in plasma membrane NADH-ferricyanide reductase activity. These systems were examined to determine if there exists a correlation between the regulation of the PMOR system and extracellular Superoxide radical formation as measured with the fluorescence probe L-012. No correlation was found between NADH-ferricyanide reductase activity and extracellular Superoxide radical production. PMOR function in cellular proliferation appears therefore not to involve extracellular Superoxide radical production.Abbreviations CoQ₁₀ coenzyme Q₁₀ - EtBr ethidium bromide - HCO-60 polyoxyethylated hydrogenated castor oil - HEPES 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid - mtDNA mitochondrial DNA - L-012 8-amino-5-chloro-7-phenylpyrido(3,4-d)pyridazine-1,4(2H,3H)dione - SOD Superoxide dismutase     

Avoidance Responses to Infrasound in Downstream Migrating European Silver Eels, Anguilla anguilla

In an attempt to develop an efficient acoustic fish fence, we have designed an infrasound source able to generate large nearfield particle acceleration. The source generates water movements by means of two symmetrical pistons in an air-filled cylinder with 21cm bore. The pistons are driven by eccentric coupling to an electric motor, with 5cm p.p. amplitude. The piston movements are 180° out of phase. The piston reaction forces are thus opposed, leading to vibration free operation. The submergible infrasound source is operated freely suspended in the water mass. The emitted sound frequency is 11.8Hz. The particle acceleration is about 0.01ms^–2 at a distance of 3m, corresponding to the threshold intensity for deterring effects of infrasound on Atlantic salmon smolts. The sound source was employed to test the effect of intense infrasound on migrating European silver eels. Fish confined in a tank displayed startle behaviour and prolonged stress reactions, telemetrically monitored as tachycardia, in response to intense infrasound. The field tests were carried out in the River Imsa. A trap that catches all the descending eels is installed near the river mouth. The trap was separated in four equal sections. During the periods with infrasound exposure, the proportion of silver eels entering the section closest to the sound source was reduced to 43% of the control value. In the section closest to the opposite river bank, infrasound increased the proportion of trapped eels to 144% of the control values. This shift of the migrating eels away from the infrasound source was highly significant.     

Mutants in yeast affecting ethidium bromide induced rho- formation and their effects on transmission and recombination of mitochondrial genes.

Summary A series of mutants called ebi, less inducible by ethidium bromide than the parental strain for the ^+– mutation have been isolated after E.M.S. mutagenesis. Some of the ebi mutants also show an important accumulation of ^– cells, in the absence of ethidium bromide. Ebi mutations are nuclearly inherited as shown by meiotic segregation. The effects of these mutants on the transmission and recombination of mitochondrial genes among the diploid progeny of crosses have been studied. Some of the ebi mutants show a non coordinated transmission of the oli1 mitochondrial marker with respect to other mitochondrial markers unexpected for homosexual crosses. This bias which is independent from will be discussed in relation to the segregation and recombination. No significant decrease of the frequency of recombinants has been detected.Abbreviations E.B. Ethidium bromide - E.M.S. Ethyl méthane sulfonate - C^S/C^R Allelic forms of the rib 1 locus conferring chloramphenicol sensitivity/resistance - E^S/E^R Allelic forms of the rib 3 locus conferring erythromycine sensitivity/resistance - O^R/O^R Allelic forms of the oli 1 locus conferring oligomycin sensitivity/resistance - P^S/P^R Allelic forms of the par 1 locus conferring paromomycine sensitivity/resistance - D^S/D^R Allelic forms of the diu 1 or diu 2 loci conferring diuron sensitivity/resistance - C^S/C^R Allelic forms of the mitochondrial locus - ⁺ grande or respiratory competent cells - ^– petite or cytoplasmic respiratory deficient cells     

Proteins main-chain atomic displacements and density of stabilizing interactions

The assumption that interactions like hydrogen bonds, that establish the secondary structure of proteins, modulate the local flexibility of the polypeptide chain suggests a phenomenological relation between ²-the X-ray determined variance of the thermally-distributed location of a main-chain atom and the value of — a properly-defined linear density of stabilizing interactions at that location. The functional relation ² 1/ is verified from the data of lysozyme. is constructed from first principles after assuming that the thermal motions of an unstabilized polypeptide chain resemble those of a random chain. Taking the locations and the identity of the stabilizing interactions from literature, the predicted 1/g9 for lysozyme and metmyoglobin is compared with the observed ² along the proteins main-chain. The satisfactory results are discussed in the light of the possible role that hydrogen bonding plays in determining the equilibrium and the dynamic properties of the main-chain structural fluctuations in proteins, and its modelling by using a simplified mechanistic approach.     

Localization of a cruciform cutting endonuclease to yeast mitochondria

We have found a cruciform cutting endonuclease in the yeast, Saccharomyces cerevisiae, which localizes to the mitochondria. This activity apparently is associated with the mitochondrial inner membrane since the activity is not released into solution by osmolysis, in contrast to the matrix enzyme, isocitrate dehydrogenase. The cruciform cutting activity appears to be encoded by CCE1. This gene has been shown to encode one of the major cruciform cutting endonucleases present in a yeast cell. In ccel strains, which lack CCE1 endonuclease activity, the mitochondrial cruciform cutting endonucleolytic activity is also absent. Since CCE1 is allelic to MGT1, a gene required for the highly biased transmission of petite mitochondrial DNA in crosses between ⁺ and hypersuppressive ^– cells, it seems likely that the CCE1 endonuclease functions within mitochondria.     

Methods for measuring the properties of the turbulence in bubble flow

Summary A constant temperature hot film anemometer has been used to evaluate mean liquid flow velocity, bubble frequency, turbulence scale and intensity, and the rate of energy dissipation by liquid phase bubble flow.Symbols M mass - L lenght - T time - a gas/liquid interfacial area L² - a=a/V_L specific gas/liquid interfacial area with regard to the volume of the liquid L^–1 - d bubble diameter L - d mean bubble diameter L - d_e dynamic equilibrium (maximum stable) bubble size L - d_p primary bubble diameter L - d_s Sauter bubble diameter L - E specific energy dissipation rate with regard to the volume of the liquid ML^–1T^–3 - E V_L energy dissipation rate ML²T^–3 - E=E/ since =1 g cm^–3, E has the same numerical value as E. Therefore, the symbol E is used everywhere in the present paper for E and called energy dissipation rate (S. s^–2=Stokes. s^–2) L²T^–3 - E_G or _G local relative gas hold up L²T^–3 - f() autocorrelation function [Eq. (10)] L²T^–3 - f(r) cross correlation function [Eq. (11)] L²T^–3 - g acceleration of gravity LT^–2 - k constant LT^–2 - k_L mass transfer coefficient LT^–1 - k_La volumetric mass transfer coefficient with regard to the volume of the liquid T^–1 - N₀ number of crossings of u and T^–1 - n_B bubble frequency T^–1 - r distance between two points 1 and 2 of the cross correlation function L - t time T - u momentaneous liquid velocity LT^–1 - mean liquid velocity LT^–1 - mean square fluctuation velocity L²T^–2 - intensity of turbulence LT^–1 - x position coordinate L - V volume of the bubbling layer in the column L³ - V_L volume of the bubble free layer in the column L³ - V electrical voltage (in Fig. 2) L³ - v velocity scale [Eq. (6)] LT^–1 - We_crit critical Weber number [Eq. (4)] LT^–1 - w_SG superficial gas velocity LT^–1 - w_SL superficial liquid velocity LT^–1 - _G or E_G local relative gas hold up LT^–1 - smallest scale [Eq. (6)] L - time delay in the autocorrelation function [Eq. (10)] T - energy dissipation scale [E. (15)] L - _f: Taylor's vorticity scale [E. (14)] L - kinematic viscosity of the liquid L²T^–1 - density of the liquid ML^–3 - surface tension MT^–2 - dynamic pressure of the turbulence [Eq. (8)] ML^–1T^–2 - p primary (at the aerator) - e equilibrium (far from the aerator)     

Factors affecting petite induction and the recovery of respiratory competence in yeast cells exposed to ethidium bromide

Summary When growing cultures of S. cerevisiae are treated with high concentrations of ethidium bromide (>50 g/ml), three phases of petite induction may be observed: I. the majority of cells are rapidly converted to petite, II. subsequently a large proportion of cells recover the ability to form respiratory competent clones, and III. slow, irreversible conversion of all cells to petite. The extent of recovery of respiratory competence observed is dependent on the strain of S. cerevisiae employed and the temperature and the carbon source used in the growth medium. The effects of 100 g/ml ethidium bromide are also produced by 10 g/ml ethidium bromide in the presence of the detergent, sodium dodecyl sulphate, and recovery is also observed when cells are treated with 10 g/ml ethidium bromide under starvation conditions. Genetic analysis of strain differences indicates that a number of nuclear genes influence petite induction by ethidium bromide.In one strain, S288C, petite induction by 100 g/ml ethidium bromide is extremely slow under certain conditions. Mitochondria isolated from S288C lack the ethidium bromide stimulated nuclease activity found in D243-4A, a strain which shows triphasic kinetics of petite formation. This enzyme may, therefore, be responsible for the initial phase of rapid petite formation.     

Attraction of Aedes aegypti II. Velocity of reaction to host with and without additional carbon dioxide

The velocity with which Aedes aegypti (L.) reacted to odors from a human arm was the same with and without additional carbon dioxide, and was found to increase asymptotically. The derivative dy/dx=–bX_ln described the velocity of response and was not different from the derivatives of the actual data. CO₂ may exert its main effect within the central nervous system.

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Zusammenfassung Die Geschwindigkeit, mit der Aedes aegypti auf den Geruch eines menschlichen Armes reagierte, war mit und ohne Kohlendioxyd-Zusatz die gleiche und nahm asymptotisch zu. Die Ableitung dy/dx=–bX_ln beschreibt die Geschwindigket der Reaktion und weicht von der Ableitung der aktuellen Versuchsdaten nicht ab. CO₂ mag seine Hauptwirkung innerhalb des Zentralnervensystems entfalten.

     

Sequence homology and recombination between the genomes of morphologically dissimilar bacteriophages LP 52 and theta

Summary A restriction fragment map of Bacillus licheniformis temperate phage LP 52 DNA (molecular weight 38.5×10⁶) was established, using restriction endonucleases BamHI (8 target sites), BglI (10 sites), BglII (13 sites) and EcoRI (22 sites). The map is linear, with well-defined ends, without any signs of circular permutation. The DNA of a related phage, LP 51, produced identical restriction fragments. At least 62% DNA of LP 52 has been found homologous to the DNA of the recently discovered, morphologically quite dissimilar, phage , as demonstrated by hybridization of electrophoretically separated restriction fragments of DNA. Under the same conditions, the DNAs of LP 52 and of the morphologically similar Bacillus subtilis phage 105 did not cross-hybridize. The homologous regions in the genomes of phages LP 52 and have been shown to be colinear. Comparison of the cleavage maps of phages LP 52 and has shown that, within the regions of homology, not a single restriction fragment and few restriction sites have been conserved during divergent evolution. Three major regions of heterology were defined; the longest one, covering the right-hand end of the map (73±2.75% up to 100% LP 52 genome length) appeared to contain genes coding for structural proteins of the virions; a shorter region at the left-hand end of the map (coordinates zero to 10.3±3.3% LP 52 genome length) and a very short central region (coordinates 41.8–43.9%) could be identified, the latter apparently containing a regulatory locus responsible for the heteroimmune behavior of the two phages. Recombinants between phages LP 52 and were isolated. Mapping of recombinant genomes has indicated mutual substitution of allelic pieces of LP 52 and DNAs upon strict conservation of overall genome length.     

A simple method for high resolution banding of chromosomes in amniotic fluid cells

Summary A simple technique which results in good quality early mitotic stages of amniotic fluid (AF) cells is presented. Two days after trypsinization AF cell cultures are incubated for 4 h in culture medium containing 20 U/ml liquemin. During the last hour 5 g/ml ethidium bromide (EB) is added and 15 min before harvest 0.04 g/ml colcemid is applied as usual. G-banded and Q-banded chromosomes corresponding to at least 550–850 bands per haploid genome can be obtained in sufficient numbers.     

Hydrodynamic characteristics of two-phase inverse fluidized bed

Hydrodynamic characteristics of a new mode of liquid-solid fluidization, termed as inverse fluidization in which low density floating particles are fluidized with downward flow of liquid, are experimentally investigated. The experiments are carried out with low density particles (<534 kg/m³) which allow high liquid throughputs in the system. During the operation, three regimes, namely, packed, semi-fluidization and fully fluidization are encountered. Empirical correlations are proposed to predict the pressure drop in each regime. A computational procedure is developed to simulate the variation of pressure drop with liquid velocity.List of Symbols Ar modified Archimedes number, d _p ³ (– _s)g/² - d _p particle diameter, mm - f friction factor (eq. 2) - g acceleration due to gravity, m/s² - H total bed height, m - H _c height of the column, m - H_f height of fluidized bed, m - H₀ height of initial bed, m - H_p height of the packed bed, m - (p) pressure drop across the bed, N/m² - (p) _f pressure drop across fluidized bed section, N/m² - (p) _p pressure drop across the packed bed section, N/m² - (p) _sf total pressure drop in semifluidization regime, N/m² - Re Reynolds number, d _pU ₁/ - Re_m modified Reynolds number, d _pU ₁/(1– _p) - U ₁ superficial liquid velocity, m/s - U_mf minimum fluidization velocity, m/s - U_osf onset fluidization velocity, m/s Greek Letters _f voidage of fluidized bed - _p voidage of packed bed - liquid viscosity, kg/ms - liquid density, kg/m³ - _s particle density, kg/m³     

An approach for parameter estimation of biotechnological processes

An approach for parameter estimators design of biotechnological processes (BTP) is presented in case of lack of real time information about state variables. It is based on general reaction rate models and measurements of at least one reaction rate. A general parameter estimator of BTP is designed with the help of which specific rate estimators are synthesized. Stability and convergence of an estimator of specific growth rate for a class of aerobic batch processes are proved. Its effectiveness is illustrated by simulation results. The proposed on-line parameter estimation approach can be used for design of BTP on-line variable estimation algorithms (variable observers of BTP).List of Symbols X, S, P g/l biomass, substrate and product concentrations - C g/l oxygen concentration in the culture broth - C _sg/l saturation concentration of oxygen in the culture broth - C _in, C_outg/l oxygen concentrations in the input air flow and in the outlet gasphase - F _in, F_outl/h the input air flow in the fermenter and output air flow - OUR g/(lh) oxygen consumption rate - OUR _mg/(lh) measured values of OUR - V l volume - , , l/h specific growth, consumption and synthesis rates - K _La(o) l/h specific volumetric mass transfer coefficient - D l/h dilution rate - R _X, R_S, R_Pg/(lh) biomass growth, substrate consumption and product synthesis rates - K _b matrix of yield coefficients - H_b(), H() matrices of known functions of - H(R) matrix of known functions of R - and gain matrices - a vector of the state variables - () a reactions rates vector, describing qualitative relations among the components - R() a reactions rates vector, describing qualitative and quantitative relations among the components - F a feed rates vector - Q a gaseous outflow rates vector - _b () a vector of unknown functions of - ₁() a vector of functions - (t) a vector of unknown time-varying parameters - ₂(, ) an auxiliary vector-function of and - Y _X/S, Y_X/C, Y_X/P substrate, oxygen and product yield coefficients - b maintenence coefficient - k _i(i=1...6) kinetic coefficients - C _i(i=1,2) design parameters estimate     

Effect of amino acids on glutathione production by Saccharomyces cerevisiae

Summary The constituent amino acids of the glutathione (GSH) tripeptide chain, glutamate, cysteine and glycine, were investigated for positive effects on GSH production in shake-flask cultures of Saccharomyces cerevisiae with glucose as the carbon source. Cysteine was confirmed as the key amino acid for increasing the specific GSH production rate, g, but showed some growth inhibition, especially in the second growth phase (ethanol-assimilation phase). An intracellular cysteine delivery agent, thiazolidine, showed a similar pattern of increased GSH production and growth inhibition, but to a slightly lesser degree, compared with free cysteine. The initial cysteine concentration affected both the specific growth rate, µ, and g, up to about 5 mm for µ and about 2–3 mm for g. Results of the [³⁵S]cysteine-labelling experiments suggest a complicated role of cysteine in increasing GSH production and further investigation may be necessary. Offprint requests to: S. Shioya     

Comparative studies of the effects of acridines and other petite inducing drugs on the mitochondrial genome of Saccharomyces cerevisiae.

Summary The effects of the acridines euflavine and proflavine on mitochondrial DNA (mtDNA) replication and mutation inSaccharomyces cerevisiae have been compared. In contrast to previous results we found that under our conditions proflavine can indeed induce high levels (>80%) of petite mutants, although six times less efficiently than euflavine. The parameters measured for mutagenesis of the mitochondrial genome and inhibition of mtDNA replication in whole cells suggest that the modes of action of euflavine and proflavine are very similar. After extended (18h) treatment of growing cells with each drug the percentage loss of mtDNA or genetic loci was almost coincidental with the extent of petite induction.It was found that proflavine is equally as effective as euflavine in inhibiting mtDNA replication in isolated mitochondria in contrast to the differential between the drugs observed in vivo. However, proflavine and euflavine inhibit cellular growth at almost the same concentrations. It is therefore proposed that there is some intracellular permeability barrier which impedes proflavine access to the mitochondrial DNA replicating system.The petites induced by euflavine (and proflavine) are characterized by there being a preferential induction ofrho ⁰ petites lacking mtDNA as opposed torho ^- petites retaining mtDNA. This is in contrast to the relative proportions of such petites induced by ethidium bromide or berenil. A scheme for the production of petites by euflavine is presented, in which euflavine inhibits the replication of mtDNA, but does not cause direct fragmentation of mtDNA (unlike ethidium bromide and berenil). The proposed scheme explains the production of the high frequency ofrho ^o cells, as well as therho ^- cells induced by euflavine. The scheme also accounts for previous observations that euflavine only mutants growing cultures, and that the buds, but not mother cells, become petite.     

Effects ofp-chlorophenylalanine on the metabolism of serotonin from 5-hydroxytryptophan

The effects ofD,L--chlorophenylalanine methyl ester (PCPA-methyl ester) and two of its metabolites, 2-(-chlorophenyl)-ethylamine (PCPEA) and -chlorophenylacetic acid (PCPAA), on the metabolism of serotonin (5-HT) fromD,L-5-hydroxytryptophan (5-HTP) ware studied in vitro and in vivo using the telencephalon and brainstem of the rat. For in vivo studies and some in vitro experiments, rats were injected with either 100 mg/kg PCPA-methyl ester or saline alone on days 1, 2, and 3, and were killed on day 15. When the in vivo metabolism of 5-HT was to be studied, the saline group and the PCPA group of animals were injected with 75 g/kg [³H]D,L-5-HTP 20 min before sacrificing. With respect to the values found for the saline-injected animals, the specific activity (S.A.; dpm/nmol) of 5-HIAA was significantly greater in the telencephanol and brainstem of the animals injected with PCPA-methyl ester. The S.A. of 5-HTP was the same in both groups; the S.A. of 5-HT was lower in the telencephalon of the PCPA group than in the saline group; in the brainstem, there was no difference. In both the saline- and PCPA-injected animals, the S.A. of 5-HIAA was greater than the S.A. of 5-HT. There was no difference between the saline- and PCPA-injected animals with regard to: (1)L-5-HTP decarboxylase activity; (2)L-5-HTP-induced release of [³H]5-HT in vitro from crude nerve ending fractions (P₂); or (3) in vitro uptake of [³H]D,L-5-HTP and its conversion to [³H]5-HT using the P₂ fraction. In vitro studies demonstrated that the PCPEA could directly cause a large increase in the release of [³H]5-HT from the P₂ fraction, whereas PCPA and PCPAA had little or no apparent effect. The data were interpreted to suggest that in the telencephalon of the animals treated with PCPA-methyl ester, there was a higher turnover of 5-HT than was found in the saline-treated group.     

Biogenesis of mitochondria 34

Summary Mutants of the yeast Saccharomyces cerevisiae have been isolated in this laboratory which show increased resistance to a number of structurally and functionally unrelated antibiotics such as mikamycin, chloramphenicol, oligomycin and tetracycline (Bunn et al., 3971). When a multiply resistant haploid strain was crossed to an antibiotic sensitive strain, the resultant diploid progeny were completely resistant to chloramphenicol and oligomycin. However, the progeny showed different responses to mikamycin depending upon the concentration of antibiotic, all showed resistance to 25 g/ml but only about half were resistant to high levels of mikamycin (>100 g/ml). Detailed genetic analyses has shown that resistance to high levels of mikamycin is the result of a phenotypic interaction between two mutations, one nuclear and the other mitochondrial. The nuclear mutation by itself confers resistance to a number of antibiotics including chloramphenicol, oligomycin and mikamycin at a level of 25 g/ml. The mitochondrial mutation increases cellular resistance to mikamycin from 3 g/ml to about 8 g/ml. When the two mutations occur together in a cell, resistance to mikamycin is increased to at least 800 g/ml, the limit of solubility. Thus, the phenotypie interaction between these two mutations is not additive but synergistic.When cells containing the cytoplasmic [mik1-r] mutation are treated with ethidium bromide to produce ^° cells (no mtDNA), the [mik1-r] determinant is lost, indicating that this mutation is located in the mitochondrial DNA. Recombination analyses with other mitochondrial markers indicates a marker order of [oli1-r mik1-r ery1-r] with [mik1-r] showing tighter linkage to the [oli1-r] marker.     

Conservation of nonhistone chromosomal proteins through dipteran evolution

We have investigated the possibility that some high molecular weight nonhistone chromosomal (NHC) proteins may have been conserved through the evolution of two distantly related diptera-Drosophila melanogaster and Sciara coprophila. Antisera produced against three NHC protein subfractions were analyzed for cross-reactivity with Sciara polytene chromosomes. The indirect immunofluorescent staining technique used couples an assay for immunologic cross-reactivity with an assay for the in situ distribution of the proteins under study. The results indicate that the - and NHC protein antigens have been conserved since the divergence of Drosophila and Sciara, while the Drosophila NHC protein antigen is not present on Sciara chromosomes. In one case, with anti-- serum, we have identified a highly conserved, very high molecular weight NHC protein (or class of proteins) which appears to interact strongly with all chromatin in a manner which is not DNA sequence-specific. In the second case, with anti- serum, we have identified an NHC protein which may have evolved an additional function(s) in Sciara relative to its function(s) in Drosophila.     

Testing the “methanochondrion concept”: are nucleotides transported across internal membranes in Methanobacterium thermoautotrophicum?

In order to test the Methanochondrion concept, uptake of adenine nucleotides in various membrane preparations of Methanobacterium thermoautotrophicum was studied. The uptake showed properties which are in general interpreted as indicative of a transport mechanism: (i) kinetics in the time range of minutes, (ii) temperature dependence, (iii) substrate specificity and (iv) failure to remove the substrate by extensive washing.However, nucleotide transport as an interpretation of this uptake can definitely be excluded. Not only an exchange mechanism of the mitochondrial type, but also a general exchange or an uniport mechanism was ruled out. In contrast, the nucleotide uptake was shown to be actually a tight and specific binding of ADP and ATP to binding sites at the interior side of the cell membrane. This was conclusively demonstrated in protoplasts obtained from M. thermoautotrophicum cells. In these protoplasts which do not contain internal membranes also nucleotide binding was observed, but only after disruption of the plasma membrane by osmotic lysis, which leads to the exposure of binding sites.     

Vascular dynamics

A study, using the four-electrode impedance plethysmograph system, was completed to evaluate simultaneous variations in conduction of upper and lower body segments relative to displacement of blood volume during change in body position. Measurements of cardiac output were compared with simultaneous results by dye dilution methods as a means of assessing the use of impedance techniques to determine cardiac output during tilt table studies. Two groups, 48 healthy private pilots and 22 patients with diabetes mellitus, were tested and the results were compared.Control and test heart rate values were higher in the afternoon than in the morning for the same healthy subjects, and the blood pressure and heart rate changes paralleled the variations in stroke volume and calf blood pulse changes. The results in the patients with diabetes differed markedly in terms of the magnitude of the cardiovascular changes and indicated the value of the tilt table in assessing fatigue in the circulatory system as a result of metabolic disturbance. The change from horizontal to 65 degree head up position in the patients with diabetes showed a marked fall in thoracic stroke and conductive volume in contrast to the minimal decrease observed in healthy subjects.Symbols resistivity, ohm cm - L length or distance between detecting electrodesE ₁-E ₂, cm - E voltage, volts - I current, amps - R ₀ segmental resistance between detecting electrodesE ₁-E ₂, ohms - V ₀ segmental volume equivalent to (L ²/R ₀), ml - V change in volume, ml - R change in resistance, ohms Terms Impedance plethysmography Measurement of change in volume due to variation in electrical resistance of a segment to a 50 or 120 kHz signal. The impedance at these frequencies is primarily resistive - Four electrodes Two electrodes for introduction of the reference signal to the examined segment and two electrodes for detecting variation in conduction of the signal - Conduction The reciprocal of resistance measured in mohs. - Conductive volume The volume defined byV ₀=(L ²/R ₀) containing electrolytes including whole blood and plasma This project was supported by Contract NAS4-1321, NASA Flight Research Center, Edwards, California, U.S.A.