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     

Membrane pathways for water and solutes in the toad bladder: II. Reflection coefficients of the water and solute channels

Summary Urea and water transport across the toad bladder can be separately activated by low concentrations of vasopressin or 8 Br-cAMP. Employing this method of selective activation, we have determined the reflection coefficient () of urea and other small molecules under circumstances in which the bladder was transporting urea or water. An osmotic method for the determination of was used, in which the ability of a given solute to retard water efflux from the bladder was compared to that of raffinose (=1.0) or water (=0). When urea transport was activated (low concentration of vasopressin), for urea and other solutes was low, (_urea,0.08–0.39;_acetamide, 0.55; _{ethylene glycol}, 0.60). When water transport was activated (0.1mm 8 Br-cAMP) _urea approached 1.0 _urea also approached 1.0 at high vasopressin concentrations. In a separate series of studies, _urea was determined in the presence of 2×10^–5 m KMnO₄ in the luminal bathing medium. Under these conditions, when urea transport is selectively blocked, _urea rose from a value of 0.12 to 0.89. Thus, permanganate appears to close the urea transport channel. These findings indicate that the luminal membrane channels for water and solutes differ significantly in their dimensions. The solute channels, limited in number, have relatively large radii. They carry a small fraction (approximately 10%) of total water flow. The water transport channels, on the other hand, have small radii, approximately the size of a water molecule, and exclude solutes as small as urea.     

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.     

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     

Free sigma subunit of Bacillus subtilis RNA polymerase binds to DNA

Summary The affinity of Bacillus subtilis RNA polymerase and subunits to DNA was examined by a non-denaturing polyacrylamide slab gel electrophoresis method which made it possible to resolve DNA-bound and free subunits. The results revealed that subunit, but not subunit had a relatively high affinity for double stranded DNA. The subunit was bound maximally to super-coiled pGR1-3 plasmid DNA at a mass ratio of /DNA of 0.7. With B. subtilis double stranded linear DNA one subunit was bound per approximately 1,000 base pairs. The -DNA complex was sufficiently stable for isolation by a molecular gel filtration column. The subunit had much higher affinity for super-coiled than for linear pGR1-3 DNA or for linear double stranded or denatured DNA from B. subtilis, E. coli, and calf thymus. These results indicate that the free B. subtilis subunit, in contrast to the E. coli subunit, can bind by itself to DNA.     

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)     

Converging catabolism of 2,4,6-trinitrophenol (picric acid) and 2,4-dinitrophenol by Nocardioides simplex FJ2-1A

Initial F₄₂₀-dependent hydrogenation of 2,4,6-trinitrophenol(picric acid) generated the hydride -complex of picrate and finally the dihydride complex.With 2,4-dinitrophenol the hydride -complex of 2,4-dinitrophenolis generated. The hydride transferring enzyme system showed activity against several substituted2,4-dinitrophenols but not with mononitrophenols. A K_m-value of0.06 mM of the hydride transfer for picrate as substrate was found. The pH optimaof the NADPH-dependent F₄₂₀ reductase and for the hydride transferase were 5.5and 7.5, respectively. An enzymatic activity has been identified catalyzing the releaseof stoichometric amounts of 1 mol nitrite from 1 mol of the dihydride -complexof picrate. This complex was synthesized by chemical reduction of picrate and characterizedby ¹H and ¹³C NMR spectroscopy. The hydride -complex of 2,4-dinitrophenolhas been identified as the denitration product. The nitrite-eliminating activitywas enriched and clearly separated from the hydride transferring enzyme system byFPLC. 2,4-Dinitrophenol has been disproven as a metabolite of picrate (Ebert et al. 1999)and a convergent catabolic pathway for picrate and 2,4-dinitrophenol with thehydride -complex of 2,4-dinitrophenol as the common intermediate has been demonstrated.     

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 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     

Precision of neural timing: effects of convergence and time-windowing

We study the improvement in timing accuracy in a neural system having n identical input neurons projecting to one target neuron. The n input neurons receive the same stimulus but fire at stochastic times selected from one of four specified probability densities, f, each with standard deviation 1.0 msec. The target cell fires if and when it receives m inputs within a time window of msec. Let _n,m, denote the standard deviation of the time of firing of the target neuron (i.e. the standard deviation of the target neuron's latency relative to the arrival time of the stimulus). Mathematical analysis shows that _n,m, is a very complicated function of n, m, and . Typically, _n,m, is a non-monotone function of m and and the improvement of timing accuracy is highly dependent of the shape of the probability density for the time of firing of the input neurons. For appropriate choices of m, , and f, the standard deviation _n,m, may be as low as . Thus, depending on these variables, remarkable improvements in timing accuracy of such a stochastic system may occur.     

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.     

An experimental approach to evaluation of the relative hydrophobic character of biological liquids and tissues

Summary Total proteins extractable from a number of rat tissues were partitioned in the aqueous Ficoll-400-Dextran-70 biphasic system. The partition coefficient of the total proteins extractable from a given tissue, K, was shown to be a tissue-specific constant. The free energy of transfer of a CH₂ group from water to the aqueous solution of a given protein extract, (gCH₂), was calculated from the corresponding K-value using the correlation relationship between K and (gCH₂) reported earlier. It is suggested to use the parameter (gCH₂)-value as a measure of the relative hydrophobic character of biological fluids and tissues. The difference between the (gCH₂)-values for blood plasma medium and for a given tissue medium is used to quantify the difference in the relative hydrophobic character of the biological tissues and fluids under study. The possibilities to use the above characteristics in drug research are considered.     

A method for the estimation of χ1 torsion angles in proteins

Summary A method for estimating CH-CH coupling constants from the shape and fine structure of NH-CH fingerprint-region cross peaks of COSY spectra is presented. Spectral simulations have been used to analyse the effect of variations in ³J_NH-CH, ³J_CH-CH, linewidths and digital resolution on the appearance of NH-CH COSY cross peaks. On the basis of these simulations a set of rules for broadly categorising experimental NH-CH cross peaks according to CH-CH coupling constants has been devised. The method has been applied to the analysis of NH-CH cross peaks of hen lysozyme. The results are compared to previous measurements of CH-CH coupling constants using E.COSY techniques.     

Observations on phase-locking within the response of primary muscle spindle afferents to pseudo-random stretch

In order to uncover encoder properties of primary muscle spindle afferent fibers, time coupling (phase-locking) of action potentials on cyclic muscle stretch was studied by means of pseudo-random noise. In cats Ia action potentials were recorded from dorsal root filaments and the gastrocnemius muscles of one hind leg were stretched. The stimulus time course was a determined sequence of randomly varying muscle length which could be applied repeatedly (sequence duration 0.6 or 20 s). The noise amplitude (standard deviation of displacements) was varied between 5 and 300 m, the upper cut-off frequency of noise f _c was varied between 20 and 100 Hz. The responses to the consecutive pseudo-random noise cycles were displayed as raster diagrams and cycle histograms. Phaselocking characterized the responses at all noise amplitudes outside the near threshold range (>10 m). The higher and f _c , the stronger was the phase-locking of impulses on the stretch. When and f _c were selected to achieve high mean stretch velocities of about 500 mm/s, phase-locking was as precise as 0.15 ms, measured as the variability of spike occurrences with respect to stretch. The rasters obtained with low noise amplitudes (<40 m) showed a loose phase-locking and this gave insight into underlying mechanisms: The elicitation of action potentials caused by dynamic stretch can be prevented by a post-spike depression of excitability. This disfacilitation was very effective in counteracting weak stretch components within the random sequence and less effective or even missing when relatively strong stretch components could force the spike elicitation. This led to the reestablishment of phase-locked patterns. The results were discussed in relation to the known encoder models.     

Acute effects of σ ligands on the extracellular DOPAC level in rat frontal cortex and striatum

Acute administration of (+)-N-allylnormetazocine ((+)-SKF-10,047) and (±)-pentazocine, was found to increase the extracellular level of 3,4-dihydroxyphenylacetic acid (DOPAC), a major dopamine (DA) metabolite, in the rat frontal cortex. By contrast, these benzomorphan ligands did not change the extracellular DOPAC level in the rat striatum. On the other hand, 1,3-di(2-tolyl)guanidine (DTG) increased the extracellular DOPAC level in the frontal cortex, while it decreased that level in the striatum. Another non-benzomorphan ligand, (+)-3-(3-hydroxyphenyl)-N-(1-propyl)piperidine ((+)-3-PPP) decreased the extracellular DOPAC level in both frontal cortex and striatum. Moreover, the increase of the extracellular DOPAC level elicited by (+)-SKF-10,047 was significantly inhibited by rimcazole, a putative antagonist, while the DTG-induced increment was not reversed by rimcazole. These findings indicated that the effects of ligands on the mesocortical DA neurons differed from those on the nigrostriatal DA neurons. In addition, the effects of benzomorphan ligands on the central DA neurons were different from those of non-benzomorphan ligands.     

σ1 Receptor Subtype Is Involved in the Facilitation of Cortical Dopaminergic Transmission in the Rat Brain

Our previous studies have shown that three sigma () receptor ligands, (+)-N-allylnormetazocine ((+)-SKF-10,047), (±)-pentazocine and 1,3-di(2-tolyl)guanidine (DTG) differently regulated the dopamine (DA) transmission in the rat brain. In the present study, we attempted to clarify the role of ₁ receptor subtype in the regulation of DA transmission using a novel and selective ₁ receptor agonist, 1-(3,4-dimethoxyphenethyl)-4-(3-phenylpropyl)piperazine dihydrochloride (SA4503) in the rat brain. Acute administration of SA4503 (1.0 mg/kg, p.o.) significantly increased DA and 3,4-dihydroxyphenylacetic acid (DOPAC) levels in the rat frontal cortex, but not in the other six regions, hippocampus, striatum, midbrain, cerebellum, medulla/pons and hypothalamus. The increase of cortical DA level elicited by SA4503 was fully reversed by N,N-dipropyl-2-(4-methoxy-3-(2-phenylethoxy)phenyl)ethylamine (NE-100) (0.25 mg/kg, p.o.), a putative ₁ receptor antagonist. In addition, SA4503 (1.0 mg/kg, p.o.) showed an increase of cortical L-3,4-dihydroxyphenylalanine (L-DOPA) accumulation under the inhibition of dopa decarboxylase activity with m-hydrobenzylhydrazine (NSD-1015), suggesting that SA4503 has activated the cortical DA synthesis rate. These results suggest that the ₁ receptor subtype plays an important role in the facilitation of cortical DA transmission. In addition, this phenomenon is partially involved in the augmentation of DA synthesis rate.     

Correlation Relation for the Membrane Transport ParametersLp, σ, and ω

We derive a formula for the correlation of the three practical transportparameters L_p, , and appearing in Kedem-Katchalskyequations. It has a form = KL_p/v_s(1-), where K = 0.0306 is a universal constant independent ofthe choice of a membrane and a solute. It can be used to calculate the valueof any of these parameters, provided the other two and the molar volume of the solute are known. The formula couldbe very useful, in particular when measurements of the parameters aredifficult or even impossible.     

The molecular basis of HbH disease in Taiwan

Summary We have determined the molecular characteristics of -thalassemia in 12 HbH subjects from Taiwan by restriction endonuclease mapping with -and -specific probes. We have found four types of defects in the -thalassemia-2 genetic determinant: ^3.7 type I; ^4.2; ^CS; and ^T. All HbH subjects carried the ——^SEA genotype in the -thalassemia-1 determinant. At least two different subtypes of ——^SEA genotype were observed in this study.