Foam behavior of biological media

Summary The surface tension and foaminess of (a) unlimited, (b) substrate limited, and (c) oxygen transfer limited growth media of Hansenula polymorpha were measured using methanol, ethanol or glucose as a substrate.The time dependence of can be described by the Avrami-Überreiter relationship: log (2.3 log V)=n log t+log b, where V = (_O – _eq/(_t – _eq, and _O, _t and _eq are at t_M=0, t_M=t and t_M (equilibrium value).The constants n and b are functions of the fermentation time t_F as long as the growth is unlimited but they are constant in the state of limited growth. With glucose substrate, the foaminess can be presented as a definite function of the time, t_DG, which is necessary to attain _eq. With alcohol as a substrate no definite (t_DG) function was found.Symbols b constant in Eq. (1) - n constant in Eq. (1) - S substrate concentration - T temperature - t_M time h (measured from the beginning of the determination of the surface tension ) - t_F cultivation time h (measured from the time of inoculation) - t_DG time (min) necessary to attain the equilibrium surface tension ) - X dry biomass concentration (gl^–1) - V (_O – _eq)/(_t – _eq) - V_S equilibrium volume of the foam (cm³) - V_G volumetric gas flow rate during the estimation of (cm³ s^–1) - vvm volumetric gas flow rate with regard to the volume of the medium (min^–1) - w_SG superficial gas velocity (cm s^–1) - _m maximum specific growth rate (h^–1) - V_S/V_G foaminess (s) - surface tension, mMm^–1 (milli Newton m^–1) - _O at t_M=0 - _eq equilibrium surface tension ( at t_M) - _t at t_M=t - HP probes from Hansenula polymorpha cultivation - NLG non limited growth - OTLG oxygen transfer limited growth - SLG substrate limited growth     

Electric field-induced breakdown of lipid bilayers and cell membranes: A thin viscoelastic film model

Summary A simple viscoelastic film model is presented, which predicts a breakdown electric potential having a dependence on the electric pulse length which approximates the available experimental data for the electric breakdown of lipid bilayers and cell membranes (summarized in the reviews of U. Zimmermann and J. Vienken, 1982,J. Membrane Biol. 67:165 and U. Zimmermann, 1982,Biochim. Biophys. Acta 694:227). The basic result is a formula for the time of membrane breakdown (up to the formation of pores): =(/C)/( _m ² ₀ ² U ⁴/24Gh ³+T ²/Gh–1), where is a proportionality coefficient approximately equal to ln(h/2₀),h being the membrane thickness and ₀ the amplitude of the initial membrane surface shape fluctuation ( is usually of the order of unity), represents the membrane shear viscosity,G the membranes shear elasticity modules, _m the membrane relative permittivity, ₀=8.85×10^–12 Fm,U the electric potential across the membrane, the membrane surface tension andT the membrane tension. This formula predicts a critical potentialU _c ;U _c =(24Gh ³/ _m ² ₀ ² )^1/4 (for = andT=0). It is proposed that the time course of the electric field-induced membrane breakdown can be divided into three stages: (i) growth of the membrane surface fluctuations, (ii) molecular rearrangements leading to membrane discontinuities, and (iii) expansion of the pores, resulting in the mechanical breakdown of the membrane.     

Theoretical studies on the necessary number of components in mixtures

Summary Theoretical studies on the optimal numbers of components in mixtures (for example multiclonal varieties or mixtures of lines) have been performed according to phenotypic yield stability (measured by the parameter variance). For each component i, i = 1, 2,..., n, a parameter u_i with 0 u_i 1 has been introduced reflecting the different survival and yielding ability of the components. For the stochastic analysis the mean of each u_i is denoted by u ₁ and its variance by _i ² For the character total yield the phenotypic variance V can be explicitly expressed dependent on 1) the number n of components in the mixture, 2) the mean of the _i ² 3) the variance of the _i ² 4) the ratio and 5) the ratio _i ² /² where denotes the mean of the u _i and _u ² is the variance of the u _j. According to the dependence of the phenotypic stability on these factors some conclusions can be easily derived from this V-formula. Furthermore, two different approaches for a calculation of necessary or optimal numbers of components using the phenotypic variance V are discussed: A. Determination of optimal numbers in the sense that a continued increase of the number of components brings about no further significant effect according to stability. B. A reduction of b % of the number of components but nevertheless an unchanged stability can be realized by an increase of the mean of the u _i by 1% (with and _u ² assumed to be unchanged). Numerical results on n (from A) and 1 (from B) are given. Computing the coefficient of variation v for the character total yield and solving for the number n of components one obtains an explicit expression for n dependent on v and the factors 2.-5. mentioned above. In the special case of equal variances, _i ² = _o ² for each i, the number n depends on v, x = (₀/)² and y = (_u/)². Detailed numerical results for n = n (v, x, y) are given. For x 1 and y 1 one obtains n = 9, 20 and 79 for v = 0.30, 0.20 and 0.10, respectively while for x 1 and arbitrary y-values the results are n = 11, 24 and 95.This publication is an extended version of a lecture given at the 1984-EUCARPIA meeting (Section Biometrics in Plant Breeding) in Stuttgart-Hohenheim (Federal Republic of Germany)     

Probabilities of negative estimates of genetic variances

Summary The probability of negative analysis of variance estimates of genetic variance components due to sampling error (Ps) was investigated. The objectives were to evaluate the magnitude of Ps, to compare Ps for estimates of _A ² and _D ² , and to compare Ps for genetic variance component estimates from the nested and factorial mating designs. Ps was defined in terms of ratios of mean squares and the F distribution was used to calculate probabilities of the negative estimates. The results indicated that Ps is often greater than 0.20 for _D ² . It is generally lower for _A ² than for _D ² , and lower for the factorial mating design than the nested mating design.Technical Contribution No. 2589 from the South Carolina Agricultural Experiment Station, Clemson University     

A method for the estimation of gene flow parameters from a population structure caused by restricted gene flow and genetic drift

Summary A method has been developed which enables the estimation of the plant gene flow parameters _p (pollen dispersal), _s (seed dispersal) and t (outcrossing rate) from a selection-free continuously structured population in equilibrium. The method uses Wright's F-coefficients and introduces a new F-function which describes the genetic similarity as a function of the spatial distance. The method has been elaborated for wind pollinated plant species but can be modified for insect pollination and for animal species. In practice allozymes will provide for the necessary neutral genetic variation. The more loci used and the more intermediate the gene frequencies, the more reliable the results. For the estimation of _p and t together (when the outcrossing rate is not known) at least two chromosomally unlinked loci are required. The method for estimating _s depends on whether the plant species is annual or perennial. The mechanism of selfing has been analysed by the explanation of the value of t by three components: population density (d), pollen flow (_p) and relative fertilization potential of own pollen (Z). The concepts of neighbourhood size and isolation by distance, developed by Wright, who used a single gene flow parameter , have been extended to the situation which is realistic for seed plants, using all three parameters _p, _s and t. When _p is large with respect to _s, _s largely determines the value of the neighbourhood size, whereas _p is the most dominating factor in isolation by distance. The use of local effective population size and mean gene transport per generation instead of neighbourhood size and neighbourhood area, respectively, is proposed to avoid confusion. Computer simulations have been carried out to check the validity and the reliability of the method. Populations of 200 plants, using two or three loci with intermediate allele frequencies, gave good results in the calculation of _p with known value of t and of _s and N_e. With unknown t, especially with lower values of t, larger populations of at least 1,000 plants are necessary to obtain reasonably accurate results for _p and mean gene transport per generation M.Grassland Species Research Group Publication No. 81     

Genetic architecture of a heterotic cross between two populations of maize

Summary The nature and magnitude of variability in the interpopulation cross of Mezcla Amarillo Selection (MAS), an introduction from CIMMYT, Mexico, and J607, a population developed in India using indigenous, American, and Yugoslavian germplasm, were studied. Interpopulation progenies developed by following the North Carolina Design I were evaluated at two locations. The additive genetic variance component in interpopulation cross, _A(12) ² , and in one population assuming the other population as tester, _A12 ² and _A21 ² were significant for all the traits evaluated, namely ear length, ear girth, kernel rows and days to silk, with one exception. For kernel rows, the dominance variance component, _A(12) ² , was also significant but it was smaller than _A(12) ² . The variance component due to dominance X location interaction, _DL(12) ² , was significant for all traits except kernel rows. In the case of ear length and ear girth, _DL(12) ² was greater than the other components. _AL(12) ² , _AL12 ² and _AL21 ² were not significant for any trait. Expected genetic advance indicated a superiority of half-sib reciprocal recurrent selection over full-sib reciprocal recurrent selection.     

RNA polymerase sigma-related proteins in Escherichia coli: detection by antibodies against a synthetic peptide

Summary Antibodies were raised against a synthetic tetradecameric peptide with an amino acid sequence, DLIQEGNIGLMKAV, which corresponds to the most highly conserved region of bacterial RNA polymerase factors. In a Western-blot analysis of total Escherichia coli proteins, the antiserum reacted specifically with at least three proteins with apparent molecular weights of 75 kDa, 27 kDa and 23 kDa, in addition to the known factors (⁷⁰ and ³²). The majorities of ⁷⁰ and ³² were recovered as associated forms with the RNA polymerase on glycerol gradient centrifugation, while the other cross-reacting proteins were not. Unambiguous evidence was obtained which indicated that the intracellular level of ³² increased rapidly upon heatshock, at least in the strain containing high copy numbers of the rpoH gene.     

Amber mutations in the structural gene for RNA polymerase sigma factor of Escherichia coli

Summary Amber mutants of Escherichia coli K-12 affected in the structural gene (rpoD) for th subunit of RNA polymerase have been obtained from a strain harboring a temperature-sensitive amber suppressor (supF-Ts6) which is active only at low temperatures. These mutants grow normally at low temperature (30°C) but do not grow at high temperature (42°C) due to the inability to synthesize factor. In one mutant studied in detail (rpoD40), the rate of -factor synthesis at 30°C is about half that of the wild type and is decreased to 10%–15% within 1 h of incubation at 42°C. The synthesis of core polymerase subunits or bulk protein is virtually unaffected at least for 2 h. The defect of the mutant in synthesis and growth at high temperature can be suppressed by any of the amber suppressors tested (supD, supE or supF). RNA-polymerase holoenzymes prepared from the mutant cells carrying each of the suppressors (grown at 42°C) exhibit different thermostabilities attributable to alterations in the factor. The reduced synthesis in the mutant is accompanied by the synthesis of polypeptide tentatively identified as amber fragment. These results as well as the genetic mapping data indicate that the amber mutation (rpoD40) resides within the structural gene for the factor and directly affects synthesis upon inactivation of the suppressor at high temperature.     

Relationships between genotype x environment interactions and rank orders for a set of genotypes tested in different environments

Multilocation trials in plant breeding lead to cross-classified data sets with rows=genotypes and columns=environments, where the breeder is particularly interested in the rank orders of the genotypes in the different environments. Non-identical rank orders are the result of genotype x environment interactions. Not every interaction, however, causes rank changes among the genotypes (rank-interaction). From a breeder's point of view, interaction is tolerable only as long as it does not affect the rank orders. Therefore, the question arises of under which circumstances does interaction become rank-interaction. This paper contributes to our understanding of this topic. In our study we emphasized the detection of relationships between the similarity of the rank orders (measured by Kendall's coefficient of concordance W) and the functions of the diverse variance components (genotypes, environments, interaction, error). On the basis of extensive data sets on different agricultural crops (faba bean, fodder beet, sugar beet, oats, winter rape) obtained from registration trials (1985–1989) carried out in the Federal Republic of Germany, we obtained the following as main result: W ₂ ^g /( ₂ ^g + ₂ ^v ) where ₂ ^g =genotypic variance and ₂ ^v = ₂ ^ge + ₂ ^o /L with ₂ ^ge =interaction variance, ₂ ^o =error variance and L=number of replications.     

A comparative study of variation in codon 33 of the<Emphasis Type="Italic"> rpoS</Emphasis> gene in<Emphasis Type="Italic"> Escherichia coli</Emphasis> K12 stocks: implications for the synthesis of σ<Superscript>s</Superscript>

The Escherichia coli rpoS gene encodes an RNA polymerase sigma factor (sigma S or ^S) required for the expression of stationary-phase genes. In the first published rpoS sequence from E. coli K-12 codon 33 is given as CAG. However, several subsequent independent studies found the amber codon TAG at this position ( rpoSAm). Besides this amber codon, other codons such as TAT have also been found at this location in rpoS. Comparative genome analysis now leads us to propose TAG as the parental codon 33 in rpoS in E. coli K-12. Five different stocks of the strain W3110, which differ in the levels of ^S protein they express, were investigated. We sequenced the rpoS gene from these, and found a T at nucleotide position 97 in four out of the five stocks and a G at position 99 in three out of the five. W1485, a parental strain of W3110, and W3350, a derivative of W3110, are also rpoSAm mutants. Such rpoSAm mutants would be expected to show no RpoS activity. The retention of partial or intermediate ^S activity by suppressor-free rpoSAm mutants is therefore puzzling. We propose that a functional, N-terminally truncated, ^S (1–53^S) can be translated from a Secondary Translation Initiation Region (STIR) located downstream of the amber codon 33. It has recently been reported that a fragment of RpoS (1–53^S) that lacks the first 53 amino acids is functional when synthesized in vivo. Taken together, our results support the hypothesis that the original codon 33 of the rpoS gene in E. coli K-12 strains is the amber codon TAG.Communicated by W. Goebel     

Selection studies in sugarcane (Saccharum sp. hybrids)

Summary An approximate method to determine sample size for the estimation of population variance, ², is given. The estimate of ² is denoted as s² . Based on the assumption of a normal distribution for (s²/²–1), the sample size is approximately equal to 20,000 z² _p,/k²; where z is a standard normal deviate, p is the probability that s² ( 100¦s² – ²¦/²) is less than, or equal to, a critical value k, and k (measured as gDs²) is the desired precision of s² .The expected value of s², with respect to sample size, and the expected cumulative frequencies of s² over sample size for various k values are given. Their goodness of fit to the observed results was satisfactory except for populations that were different from normal. The observed values were taken from a study on four yield components in five sugarcane polycross progenies, grown in two contrasting environments over 2 years in three selection stages.The expected s² was found to be independent of the population coefficient of variance.Research suppoted in part by USDA, ARS, grant #12-14-5001-34. Published with the approval of the Director as Paper No. 412 in the Journal Series of the Experiment Station, Hawaiian Sugar Planters' Association.     

Surface charge density estimation by 9-aminoacridine fluorescence titration: improvements and limitations

A large number of surface charge density () and surface potential (_o) estimations have been based on 1) titrations of the fluorescence of 9-aminoacridine released from the diffuse double layer adjacent to negatively charged membrane surfaces by non-adsorbing monovalent and divalent cations, and 2) calculations using experimental data from the titration curves and the Gouy-Chapman theory of the diffuse double layer. In this paper we discuss the different simplifying approximations employed in the earlier calculations and recommend modified formulas for the calculations. The latter have been derived without any simplifying approximation concerning the ionic (electrolyte) composition of the titration assays. We also show that depends, to some extent, on the concentrations of buffer and vesicles in the assays and present experimental evidence that decamethonium (decane-1,10-bis-trimethylammonium), a bulky organic divalent cation, can be satisfactorily used for the estimation of under well-defined conditions, despite its putative interaction with membranes.Abbreviations 9-AA 9-aminoacridine - (DeM)²⁺ decamethonium - (DiM)²⁺ dimethonium - EDTA ethylenediaminetetraacetic acid - EGTA ethylene glyol-bis(-aminoethyl ether) N,N,N,N-tetraacetic acid - (HeM)²⁺ hexamethonium - MES 2-(N-morpholino) ethanesulfonic acid - MOPS 4-morpholinopropanesulfonic acid - PM plasma membrane - Tris tris(hydroxymethyl)aminomethane - surface charge density - _o surface potential Correspondence to: A. Bérczi     

Respiratory activity and growth kinetics of Candida yeasts related to carbon sources and available energy

Summary Three yeasts of the genus Candida (Candida intermedia, candida lipolytica and Candida tropicalis) were cultivated batchwise on three different carbon sources: glucose, acetate, and hexadecane. Growth curves, oxygen uptake rates, CO₂ evolution rates and the amount of oxygen required for biomass production were determined. The data were compared and discussed from the point of maximum specific growth rate, maximum oxygen uptake rate, carbon conversion into CO₂ and biomass, consumption of oxygen and available energy for cell synthesis. The results indicated a relationship between _m m, Y_s, Y_O, and for different carbon sources. Y_O and were in the same order of magnitude for acetate (0.58 and 0.38 respectively) and hexadecane (0.45 and 0.40 respectively). These values were remarkably lower than those for glucose (1.26 and 0.54 respectively).Symbols av e^– Available electrons per mol of substrate (dimensionless) - E_av Energy available per mol of substrate (dimensionless) - C_d Dissimilated carbon (%) - m Maximum specific rate of oxygen uptake (mMO₂ h^–1 g^–1) - RQ CO₂ evolved per O₂ consumed - mol. wt. Molecular weight - Y_ATP Biomass mass yield based on mol of ATP generated (g) - Biomass mass yield based on available energy (g) - Y_M Biomass mass yield based on mol of organic substrate (g) - Y_O Biomass mass yield based on oxygen consumed (gg^–1) - 1/Y_O Oxygen consumed for one gram of biomass produced (gg^–1) - Y_s Biomass mass yield based on organic substrate (dimensionless) - b Reductance degree of biomass (equiv. available electrons/g atom carbon) - s Reductance degree of organic substrate (equiv. available electrons/g atom carbon) - Fraction of energy in organic substrate which is converted to biomass - b Weight fraction carbon in biomass (dimensionless) - s Weight fraction carbon in organic substrate (dimensionless) - _m Maximum specific growth rate (h^–1)     

Potential induced in a spherical cell by an intracellular point source and an extracellular point sink

Summary The potential is calculated for all time, inside and outside a spherical cell for a point source of current inside the cell and a point sink located a finite distance outside the cell. The source and sink are step functions in time. An eigenfunction expansion is obtained, valid for arbitrary =_m a/_i , where _i and _m are the conductivities inside the cell and in the membrane, respectively, a is the cell radius and the membrane thickness. For small , the eigenfunction expansion is expanded in powers of . The time dependence of the potential contains transients with two widely differing time constants =C_m a/_i, where C_m is the membrane surface capacitance, and _m=/. Closed-form expressions are obtained for the two leading terms, for small , after the rapid transient is over. The remaining time dependence is only in the potential inside the cell, and is a simple exponential increase, independent of position within the cell. It is found that the transmembrane potential is insensitive to the location of the extracellular sink at long times, but not at short times. The dependence of the potential on location of source, sink, and observer is studied for long times after the quick transients are over. A uniqueness theorem is derived for the solution to Laplace's equation for the membrane boundary condition.This work was supported in part by NSF Grant No. GB-24965. Dr. Peskoff is the recipient of NIH Special Research Fellowship No. 1F03 GM 55849-01. Mr. Ramirez is a Ford Foundation Pre-doctoral Fellow.     

Simple diagnostic statistical tests of models for DNA substitution

The accuracy of models for DNA substitution used in phylogenetic analyses is becoming more important with the increasing availability and analysis of molecular sequence data. It is natural to look for ways of improving these models, and to do this in a planned manner it is useful to be able to identify features of sequences that may not be described adequately. In this paper, I describe three statistics which may give useful diagnostic information on departures from models' predictions. The statistical distributions of these statistics are discussed and simple significance tests are derived. These tests are based on the (estimated) phylogeny of the sequences and so have the advantage of using the information contained in this tree. Examples are given of the application of the new tests to Markov chain models describing the evolution of primate pseudogene sequences and small-subunit RNA sequences.Abbreviations b(N,p) binomial distribution of N trials, each with probability p of success - m(N,p ₁,p ₂, ..., p _r ) multinomial distribution of N trials, with r possible outcomes having probabilities p ₁, p ₂, ..., p_r, respectively - N(, ²) Normal distribution with mean and variance ² - p() Poisson distribution with mean - bp base pairs - cdf cumulative distribution function - i.i.d. independent, identical distribution     

Physiological relationship between the temperate phages lambda and phi80

Summary This work deals with the ability of phage 80 to provide defective mutants of with their missing functions. Functions Involved in Recombination. As shown by others, the Int mechanism of 80 cannot excise prophage . However, 80 efficiently excises recombinants from tandem dilysogens, using its Ter mechanism. Likewise, the nonspecific mechanism Red is interchangeable between 80 and . Maturation of DNA by 80. The Ter recombinants excised by 80 from tandem dilysogens are packaged into a 80 protein coat. This contrasts with the fact, already mentionned by Dove, that 80 is extremely inefficient for packaging phage superinfecting a -lysogen. The latter result is also found when the helper phage is a hybrid with the left arm of (80hy4 or 80hy41 — see Fig. 1). However, the maturation of the superinfecting is much more efficient if the 80hy used as a helper has the att-N region of (like 80hy1). Conversely a with the att-N region of 80 (hy6 — see Fig. 1) is packaged more efficiently by 80 or 80hy4 than by 80hy1. It is suggested that the maturation of chromosome superinfecting an immune cell requires a recombination with the helper phage. Vegetative Functions. Among the replicative functoons O and P, the latter only can be supplied by 80. That N mutants are efficiently helped by 80 does not tell that 80 provides the defective with an active N product; the chromosomes are simply packaged into a 80 coat. This shows that 80 is unable to switch on the late genes of . That neither 80 nor any of the 80hy tested can provide an active N product is shown in a more direct way by their complete failure to help N ^-r₁₄; this phage carries a polar mutation which makes the expression of genes O and P entirely N-dependant. The maturation of a N ^- by 80 contrasts with the fact that mutants affected in late genes (A, F or H) are not efficiently helped by 80. This suggests that the products coded by these genes are not interchangeable between 80 and , and that packaging of DNA into 80 coats is possible but inhibited when late proteins are present in the cell. Activation of the Late Genes. Among the im ₈₀ h ⁺ hybrids tested, only 80hy41 is able to switch on the late genes of a N defective mutant. This hybrid differs from the other hybrids studied here, by the fact that it has the Q-S-R region of (see Fig. 1). The results are consistant with the view that the product of Q gene is sufficient for activating the late genes of a DNA. N would thus control the expression of late genes only indirectly by controlling the expression of gene Q (Couturier & Dambly have independantly reached the same conclusion, 1970). Furthermore the failure of 80 and of the 80hy1 and 80hy4 to activate the late genes of would imply that these phages are unable to provide an Q product active on the chromosome Reciprocally, switches on the late genes of prophage 80hy41, but not of prophages 80hy1 and 80hy4. This suggests that the initiation of late genes expression takes place at a main specific site located in the Q-S-R region of the chromosome. The expression of the late genes would thus be sequential, and proceed through the left arm only when steaky ends cohere. Similar conclusions were reached independantly by Toussaint (1969) and by Herskowitz and Signer (1970).

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Ce travail a été réalisé dans le cadre du contrat d'association Euratom-U. L. B. 007-61-10 ABIB et avec l'aide du Fonds de la Recherche Fondamentale Collective.     

Dielectric properties of human blood and erythrocytes at radio frequencies (0.2–10 MHz); dependence on cell volume fraction and medium composition

The dielectric properties of human erythrocytes (red blood cells) suspended in whole blood and in isotonic media at various volume fractions (haematocrits) have been studied in the frequency range 0.2–10 MHz, in which the so-called-dispersion due to the Maxwell-Wagner effect is known to occur. The capacitance and conductance at 25 °C were measured by an instrument interfaced to a computer. The rectangular sample cavity (1 ml volume) contained four pure gold electrode pins, and the sample could be circulated by a roller pump. The frequency-dependence of the permittivity and conductivity were fitted by non-linear least squares regression. Corrections were applied for non-linearity in the dielectric increment at high haematocrit, and for electrode polarisation when diluting the blood in saline. Data were interpreted in terms of a simple equivalent resistor-capacitor circuit. From the measured haematological values the specific membrane capacitance (C_m) and the conductivities internal and external to the cells (_i and _o respectively) were estimated. The conductivities behaved in a predictable manner with a mean of 0.458 S · m^–1 (s.d. ± 0.044) for _i, whereas the value of C_m (and indeed the actual capacitance of the suspension) was dependent on the amount of plasma present. Hence, in stationary normal (anticoagulated) whole blood samples, C_m was as high as 2.98 F · cm^–2 (s.d. ± 0.40), in contrast to about 0.9 F · cm^–2 in blood diluted more than two-fold (to less than 20% hct) in isotonic media. The high value remained when the diluent was plasma. The C_m value returned to a high value when washed erythrocytes were reconstituted with plasma, provided that this was present at above a critical or threshold concentration of about 30 vol % in the medium, irrespective of the haematocrit in the range studied (15–44%). The C_m remained low in serum. When added to washed cells in saline, purified fibrinogen had no effect. However, high C_m values were obtained by fibrinogen supplementation to serum and diluted plasma. Applying moderate flow to whole blood approximately halved its high C_m value in an exponential manner with flow rate, whilst the C_m of washed cells (31–67% hct) slightly increased, and converged to the value for whole blood under flow. We interpret the highapparent C_m value in stationary samples to be a result of rapid cell aggregation in the presence of plasma, where rouleaux formation takes place before visible sedimentation sets in.     

Direct evidence of structural changes in reaction centers of Rb. sphaeroides containing suppressor mutations for Asp L213 → Asn: A FTIR study of QB photoreduction

In bacterial reaction centers (RCs), changes of protonation state of carboxylic groups, of quinone-protein interactions as well as backbone rearrangements occuring upon Q_B photoreduction can be revealed by FTIR difference spectroscopy. The influence of compensatory mutations to the detrimental Asp L213 Asn replacement on Q_B ^–/Q_B FTIR spectra of Rb. sphaeroides RCs was studied in three double mutants carrying a Asn M44 Asp, Arg M233 Cys, or Arg H177 His suppressor mutation. The proton uptake by Glu L212 upon Q_B ^– formation, as reflected by the positive band at 1728 cm^–1, is increased in the Asn M44 Asp and Arg H177 His suppressor RCs with respect to native RCs, and remains comparable to that observed in Asp L213 Asn mutant RCs. Only the Arg M233 Cys suppressor mutation affected the 1728 cm^–1 band, reducing its amplitude to near native level. Thus, there is no clear correlation between the apparent extent of proton uptake by Glu L212 and the recovery of the proton transfer RC function. In all of the mutant spectra, several protein (amide I and amide II) and quinone anion (C...O/C...C) modes are perturbed compared to the spectrum of native RCs. These IR data show that all of the compensatory mutations alter the semiquinone-protein interactions and the backbone providing direct evidence of structural changes accompanying the restoration of efficient proton transfer in RCs containing the Asp L213 Asn lesion.     

Problems in estimating growth rates of marine phytoplankton from short-term14C assays

Growth rate estimates () of phytoplankton populations that were sampled from nitrogen-limited continuous cultures and then incubated for short durations in batch culture with added¹⁴C-HCO₃ ^– were significantly different than steady-state growth rates () for 3 of 5 marine phytoplankton species. Two diatoms,Thalassiosira weissflogii andChaetoceros simplex, displayed virtually identical growth rates (=) over a wide range of, whereas for a third diatom,Phaeodactylum tricornutum, was overestimated by an average of 40% compared to. In contrast, was underestimated by the¹⁴C technique for the two remaining species: up to 40% at a steady-state of 1.0 day^–1 for the chlorophyteDunaliella tertiolecta and up to 100% at of 1.4 day^–1 for the haptophytePavlova lutheri. For the latter two species the divergence between and appeared to increase with increasing steady-state. A simple model of labeled and total carbon flow between the aqueous phase and cellular biomass was constructed to demonstrate that respiration was negligible when=, but was significant when>. In the cases in which<, a rapid physiological alteration presumably took place once the steady state was disturbed and cells were placed in the incubation chambers, which perhaps was related to the nutritional state of the cultures at the time of sampling. Questions thus are raised regarding our ability to measure accurately primary productivity from shipboard experiments with confined samples of phytoplankton from nutrient-impoverished waters that probably are less hardy than the laboratory cultures used in these studies.     

Enhancement of protein precipitate strength and density by low-frequency conditioning

The use of a continuous, low-frequency conditioning process to alter the structure of protein precipitate aggregates is examined. An increase in the density of aggregates is correlated with the levels of fluid acceleration and hence hydrodynamic stress to which the aggregates are exposed during conditioning. A combination of low-frequency conditioning followed by shear break-up (as in the feed zone to a high-speed disk-stack centrifuge) is shown to result in a precipitate suspension of increased particle size at the fine end of the distribution, and having a greater sedimentation velocity. The resistance of large aggregates to shear disruption is increased by low-frequency conditioning.List of Symbols CR conditioning ratio - CRS conditioning ratio after shearing - d m amplitude of displacement - D m particle size - D _c m critical size for centrifuge recovery - f s^–1 frequency of vibration - G s^–1 mean velocity gradient - Q m³/s volumetric throughput - SR shear ratio - t s ageing time Greek Symbols s^–1 mass-average shear rate - K sedimentation shape factor - _a kg/m³ aggregate density - _f kg/m³ fluid density - _s kg/m³ solids density - kg/m³ aggregate-suspension density difference - Ns/m² kinematic viscosity - amplitude of pulse ratio (ref. 23, 9) - s mean residence time - _s solids volume fraction