Red blood cell rouleaux formation in dextran solution: dependence on polymer conformation

The velocity of rouleaux formation (RF), as previously shown, increases with increasing dextran concentration up to a critical concentration (C_a), beyond which the addition of dextran reduces the RF velocity (RFV). de Gennes' model for polymer solutions suggests that dextrans exist in two conformations: a coil structure at low concentrations, which changes to a network beyond a critical concentration (C^*). In the present study we examined the relation between C_a and C^* for dextrans of different molecular weight, and found that they coincide. This suggests that the change in dextran behavior, from increasing to decreasing RFV, occurs when their conformation changes from coil to network. In addition, it has been reported that in dilute dextran solutions the intercellular distance (D) between RBC in rouleaux increases with the molecular weight of the dextran. We found that D correlates with R_f, the end-to-end distance of the polymer molecule, and for all dextrans D ≤ 1.5 R_f. In accord with de Gennes' Model for polymers between surfaces, this corresponds to intercellular interaction with two overlapping surface-associated polymer layers, which may extend “tails” to interact with the opposing cells. Received: 8 August 1997 / Accepted: 28 November 1997     

Analysis of the electrophoretic properties of double-stranded DNA and RNA in agarose gels at a finite voltage gradient

The electrophoretic mobilities of double-stranded (ds) DNAs and ds RNAs of various lenths, L, were measured in gels of 0.4–1.8% (w/v) agarose at a voltage gradient of 1.0 V/cm. Differences in the electrophoresis of ds DNA and ds RNA are presented and discussed. A general expression is derived that describes the electrophoretic mobility, M, of either type of ds nucleic acid as a function of the gel concentration and the nucleic acid length: M = M′₁(L/L₀)^?x ? M₂, where M′₁ and L₀ are constants, and x and M₂ depend on the agarose gel concentration. The results obtained by fitting our data with this equation are consistent with the mobilities of nucleic acids in a wide range of gel concentrations, including free electrophoresis in solution and electrophoresis in gles of high agarose concentration in which nuleic acids are expected to reptate through the gel matrix. Finally, various methods of plotting agarose gel electrophoresis data are discussed.     

Histone-induced conformational changes in DNA as probed by quasi-elastic light scattering.

Quasi-elastic light scattering as measured by intensity fluctuation (self-beat) spectroscopy in the time domain can be profitably used to follow both the translational diffusion D and the dominant internal flexing mode τ_int of DNA and its complexes with various histones in aqueous salt solutions. Without histones, DNA is found to have D = 1.6 × 10^?8 cm²/sec and τ_int ? 5 × 10^?4 sec in 0.8 M NaCl, 2 M urea at 20°C. Total histone as well as fraction F2A induce supercoiling (D = 2.6 × 10^?8 cm²/sec, τ_int ? 2.8 × 10^?4 sec) whereas fraction F1 induces uncoiling (D = 1.0 × 10^?8 cm²/sec, τ_int ? 9.4 × 10^?4 sec). Upon increasing the salt concentration to 1.5 M the DNA–histone complex dissociates (D = 1.8 × 10^?8 cm²/sec). Upon decreasing the salt concentration to far below 0.8 M, the DNA–histone complex eventually precipitates as a chromatin gel.     

Electrophoretic mobility of lambda phage HIND III and HAE III DNA fragments in agarose gels: a detailed study

The electrophoretic mobility (μ) of DNA fragments from λ phage and ΦX 174, split by restriction enzyme to molecular lengths from 3 × 10² to 2.36 × 10⁴ base pairs, has been investigated in 0.6–4% agarose gels at various field strengths, ionic strengths, and temperatures. As already observed, μ is seen to be very sensitive to the field, increasing with field strength. The sensitivity increases with the molecular length of the DNA and decreases at high gel concentration. Our data are in qualitative agreement with recent theoretical predictions that concern the influence of the electric field on electrophoretic mobility. Mobility data have been extrapolated to zero field. This enables a comparison of our experimental results with theoretical predictions on the dependence of μ on the molecular weight of the DNA fragments. Our data fit, quite closely, a reptation model, where the tube path is described as a semiflexible entity with a persistence length equal to the pore diameter. The influence of the agarose concentration and the ionic strength of the buffer on the two parameters of the model—intrinsic electrophoretic mobility (μ₀) and the number of base pairs per element of the tube (g)—are well described by the model. The temperature dependence of the electrophoretic mobility, together with the influence of the agarose concentration on μ₀, indicate that the hydrodynamic drag is the leading frictional force on the DNA molecules in the gel.     

Enzyme inactivation and stabilization studies in an ultrafiltration reactor

Summary An ultrafiltration membrane enzymatic reactor is used in connection with different reacting systems.The experimental conditions are such that the enzyme, which operates at fairly high concentration levels because of the concentration polarization phenomena taking place in the reactor, is still in soluble form.The analysis of the system unsteady-state response enables the identification of the mechanism of enzyme deactivation and the extraction of the kinetic parameters of both the deactivation and the main reaction.The stabilizing effect observed in connection with enzyme entrapment within an inert gel deposited onto the U.F. membrane active surface is also discussed.List of Symbols A U.F. membrane cross sectional area cm² - C_E Enzyme concentration mg/ml - C_EI Enzyme concentration at the active membrane surface mg/ml - C_E Mean enzyme concentration mg/ml - c _s ^o Substrate concentration in the feed m moles/ml - c _s ^u Substrate concentration in the outlet m moles/ml - D_E Enzyme diffusivity cm²/s - Km michaelis constant mM - k₂ Kinetic constant of the enzymatic reaction m moles/mg s - k_d Kinetic constant of the enzyme deactivation reaction s^–1 - N^o Initial amount of active enzyme mg - N(t) Active enzyme amount at reaction time t mg - Q Flow rate ml/s - r Rate of the main reaction m moles/ml s - t Reaction time s - t^* Reaction time at which product concentration in the outlet is within ± 2% of the steady-state value s - v Fluid velocity cm/s - V Cell volume ml - V_B Volume within which 99% of the enzyme fed is contained at the steady-state ml - V_S Volume within which 99% of the total substrate concentration drop occurs at the steady-state ml - x Distance upstream the membrane measured from the membrane surface cm     

Dipole moments of random coil polypeptides

The Rotational Isomeric States model is applied to calculate dipole moments of polypeptides of the twenty natural α-amino acids in the random coil state. Dipole moments of each repeat unit (μ_i), are evaluated using a quantum mechanics procedure. Dipole moment ratios ($\text{D}{}_{\mathrm{x}}\text{=}\text{〈μ}{}^2\text{〉}\text{x}\text{μ}{}_{\mathrm{i}}{}^2\text{,}\text{x = number of repeat units}$) of homopolypeptides are calculated and extrapolated to x →?. With a few exceptions, D_? = 0.36 ± 0.1. Ten actual proteins and three enzymes are also studied; their dipole ratios (D_x′ =〈μ〉/x) range from 7.34 to 10.57 in 10^?59 C² m² (6.6–9.5 D²). Diffferences in the values of D_x′ are due mainly to the different contributions, μ_i, of the amino acid residues contained in each polymer, whereas the sequence of amino acids has a very minor effect.     

Reproduction rate,feeding process,and liebig limitations in cell populations—II

A biological system consisting of a population of cells suspended in a liquid substrate is considered. The general problem addressed in the paper is the derivation of the kinetic pattern of population growth as a statistical effect of a very large number of elementary interactions between a single cell and the molecules of nutrient in substrate. Solution of the problem is obtained in the form of equation expressing the population growth ratec as a function of substrate concentration,C _s. The analytical expression derived is applied to a real bacterial population (Escherichi coli) and kinetic patterns are theoretically computed. The major findings, expressed roughly, without nuances, are: (i) the concentration of nutrient at the cell membrane,C _c, can only be equal to either 0 (for theC _s below some threshold valueC ^*) orC _s (forC _s>C ^*); (ii) the Michaelis-Menten-Monod kinetics observed in experiments is an artifact: the pure (not contaminated by foreign factors) dependence ofc onC _s is actually such that the functionc=c(C _s) has practically linear increase whenC _s<C ^*, and is constant,c=c(C ^*)=const, whenC _s>C ^*; (iii) the Liebig principle is strictly fulfilled: up to a feasible accuracy of observation, under no circumstances can population growth be limited (controlled) by more than one substrate component—replacement of a limiting component for another one is an instant event rather than a gradual process.     

Properties of the EEG μ rhythm and its reactivity during the performance,observation, imitation,and auditory recognition of movements in children aged 4–14 years

We studied the frequency and amplitude properties of the EEG μ rhythm in children (n = 38) aged 4–14 years under the condition of visual fixation (VF) on a video image of a computer mouse, as well as the parameters of its desynchronization during the tasks involving self-controlled performance, observation, imitation, and auditory recognition of the hand circular movements (SPM, OM, IM, and ARM, respectively) with a computer mouse. We observed age-related increment in the modal frequency of μ rhythm in loci C ₃ and C _z. During the SPM process, a significant decrease in the amplitude of μ rhythm was found in loci C ₃ (26.7%) and C _z (10.3%); during the OM, in locus C _z only (9.9%). The effects of the μ rhythm desynchronization during both self-paced and observed movements might be an evidence of well-developed neural matching mechanisms that provide the processes of performing and observing similar actions in children aged 4 to 14 years. During the IM, the desynchronization of μ rhythm was significant in loci C ₃ (27.4%) and C _z (15.3%). We hypothesize that the statistically significant increase (from SPM to IM) in the depression of μ rhythm in locus C _z is likely to be an effect of a social gaming context common for the situations when children imitate the movements of adults. The index of desynchronization of μ rhythm under the studied conditions did not depend on the age of children. Possible relations of identified modulations of the μ rhythm to the activity of the mirror neuron system were also discussed.     

Influence of base-pair changes and cooperativity parameters on the melting curves of short DNAs

Theoretical melting curves were calculated for four DNA restriction fragments, 157–257 base pairs (bp), and a series of hypothetical block DNAs with sequences d(C_2xA_xC_2x). d(C_2xT_xG_2x), 5 ? x ? 40. These DNAs provided a mixture of A·T/G·C sequence distributions with which to investigate the effects of parameters and base-pair changes on the melting of short DNAs. The sensitivity of DNA melting curves to changes in internal loop melting parameters σ and κ was examined. As Expected, theoretical melting curves of short DNAs with a quasirandom base-pair sequence vary little with changes in internal loop parameters. End melting dominates the transition behaviour of these moleucles. This was also observed for the block DNAs up to x = 22. Beyond this length, melting curves are highly sensitive to the internal loop parameters. Sensitivity is also predicted for a 157-bp fragment with a block distribution of A·T and G·C pairs. These results indicate that accurate evaluation of internal loop parameters is possible with short DNAs (100–200 bp) containing a G·C/A·T/G·C block distribution with at least 22 bp in each block. Duplex-to-single-strands dissociation parameters were reevaluated form experimental melting curve data of eight DNA fragments using a least squares fit approach. This analysis confirmed parameter values previously found with a simplified dissociation model. A Priori predictions are made on the effects of base-pair changes on the melting curves of three characterized DNA restriction fragments. Single base-pair changes are predicted to induce small but measurable changes in the melting curves. The characteristics of the altered melting curves depend on the location of the base-pair change.     

Characterisation of DNA from condensed and dispersed human chromatin

Condensed and dispersed chromatin fractions were isolated from human placental nuclei. The DNA of each fraction was purified and characterised by isopycnic centrifugation, thermal fractionation on hydroxylapatite (HAP) and sequence complexity studies. The DNAs had identical buoyant densities in neutral CsCl (1.698 g/cm³) and similar melting profiles on HAP. Analytical ultracentrifugation in Ag⁺-Cs₂SO₄, however, showed that satellite DNAs were present in the condensed fraction DNA (DNA_C) but were not visible in the dispersed fraction DNA (DNA_D). In addition, DNA_C was found to be enriched in highly reiterated sequences (20% reassociated by C₀t 10^?3) which can be correlated with the presence of satellite DNAs, whereas DNA_D contained only 3% of these fast reassociating sequences. In contrast DNA_D contained 30% intermediate sequences (reassociating between C₀t 10^?3 and C₀t 100) which represent only 10% of DNA_C. The reassociated highly repeated sequences of DNA_C showed the presence of two components in both CsCl density gradients and HAP thermal elution studies. This suggests that either there are sequence relationships resulting in partial mismatching between the different highly repeated DNA sequences in this fraction, or that highly repeated sequences are associated with less repetitious DNA. The results are discussed in terms of possible differences in genetic activity between the chromatin fractions.     

Structural and luminescence properties of Y2‐xGeMoO8:REx (RE = Eu,Tb) phosphors

Y_2‐xGeMoO₈:RE_x (RE = Eu, Tb) phosphors were synthesized using a facile sol–gel method. The morphology and structure of the phosphors were characterized by scanning electron microscopy (SEM), transmission electron microscopy (TEM) and X‐ray diffraction (XRD); while their luminescent properties were investigated by photoluminescence (PL) spectrometry. Our results reveal that all of these Y_2‐xGeMoO₈:RE_x (RE = Eu, Tb) phosphors adopted the tetragonal phase, belonging to Scheelite (CaWO₄) structure. The obtained YGeMoO₈:Eu phosphors exhibit a strong emission in the red light range which can be assigned to the ⁵D₀ → ⁷F₂ transition of Eu³⁺ when it is excited at 459 nm. Under 392 and 489 nm excitation, the YGeMoO₈:Tb phosphors present predominant green emission (⁵D₄ → ⁷F₅) at 540 nm. The highest emission of the phosphors can be achieved by adjusting the doping concentration to be 0.25 for Eu³⁺ and 0.15 for Tb³⁺, respectively. The promising luminescence properties of these materials indicate that they can be potentially applied to white‐light‐emitting diodes. Copyright © 2013 John Wiley & Sons, Ltd.     

Determination of the effective diffusion coefficient of oxygen in gel materials in relation to gel concentration

Summary The effective diffusion coefficient of oxygen, ID_e, was determined in different gel support materials (calcium alginate, -carrageenan, gellan gum, agar and agarose) which are generally used for immobilization of cells. The method used was based upon fitting Crank's model on the experimental data. The model describes the solute diffusion from a well-stirred solution into gel beads which are initially free of solute. The effect of the gel concentration on ID_e of oxygen in the gel was investigated. The results showed a decreasing ID_e for both agar and agarose at increasing gel concentration. In case of calcium alginate and gellan gum, a maximum in ID_e at the intermediate gel concentration was observed. It is hypothesized that this phenomenon is due to a changing gelpore structure at increasing gel concentrations. The ID_e of oxygen in calcium alginate, -carrageenan and gellan gum varied from 1.5^*10^–9 to 2.1^*10^–9 m²s^–1 in the gel concentration range of 0.5 to 5% (w/v).     

Assessment of protein rotational diffusion by 13C off-resonance rotating frame spin–lattice relaxation: Effect of backbone and side-chain internal motion

The ¹³C off-resonance rotating frame spin–lattice relaxation technique is applicable to the study of protein rotational diffusion behavior in a variety of experimental situations. The original formalism of James and co-workers (1978) (J. Am. Chem. Soc. Vol. 100, pp. 3590–3594) incorporated random isotropic reorientational motion of a rigid spherical rotor with no provision for backbone or side-chain carbonyl group internal motion. Here we demonstrate that the failure to include such internal motion may lead to erroneous rotational correlation time determinations for overall reorientational motion. The effect becomes severe for protein molecular masses in excess of 100 kD. Inclusion of both backbone and side-chain carbonyl carbon internal motion, using reasonable parameters derived from the literature [R. Levy and M. Karplus (1979), Chemical Physics Letters, Vol. 65, pp. 4–11; G. Careri, P. Fasella, and E. Gratton (1975), Critical Reviews in Biochemistry, Vol. 3, pp. 141–164; G. Lipari, A. Szabo, and R. Levy (1982), Nature, Vol. 300, pp. 197–198], plus corrections for anisotropic tumbling [C. F. Morgan, T. Schleich, G. H. Caines, and D. Michael (1990), Biopolymers, Vol. 29, pp. 469–480] and microscopic viscosity [S. H. Koenig (1980), ACS Symposium. Series, Vol. 127, pp. 157–176], leads to reliable values for the correlation time describing overall protein reorientation up to molecular masses of approximately 1000 kD. © 1993 John Wiley & Sons, Inc.     

Luminescence study of Eu3+doped Li6Y(BO3)3 phosphor for solid‐state lighting

In this study, Li₆Y_1–xEu_x(BO₃)₃ phosphor was successfully synthesized using a modified solid‐state diffusion method. The Eu³⁺ ion concentration was varied at 0.05, 0.1, 0.2, 0.5 and 1 mol%. The phosphor was characterized for phase purity, morphology, luminescent properties and molecular transmission at room temperature. The XRD pattern suggests a result closely matching the standard JCPDS file (#80‐0843). The emission and excitation spectra were followed to discover the luminescence traits. The excitation spectra indicate that the current phosphor can be efficiently excited at 395 nm and at 466 nm (blue light) to give emission at 595 and 614 nm due to the ⁵D₀ → ⁷F_j transition of Eu³⁺ ions. Concentration quenching was observed at 0.5 mol% Eu³⁺ in the Li₆Y_1–xEu_x(BO₃)₃ host lattice. Strong red emission with CIE chromaticity coordinates of phosphor is x = 0.63 and y = 0.36 achieved with dominant red emission at 614 nm the ⁵D₀ → ⁷ F₂ electric dipole transition of Eu³⁺ ions. The novel Li₆Y_1–xEu_x(BO₃)₃ phosphor may be a suitable red‐emitting component for solid‐state lighting using double‐excited wavelengths, i.e. near‐UV at 395 nm and blue light at 466 nm. Copyright © 2015 John Wiley & Sons, Ltd.     

Correlated motions of C′–N and C<Subscript>α</Subscript>–C<Subscript>β</Subscript> pairs in protonated and per-deuterated GB3

We investigated correlated µs-ms time scale motions of neighboring ¹³C′–¹⁵N and ¹³C_α–¹³C_β nuclei in both protonated and perdeuterated samples of GB3. The techniques employed, NMR relaxation due to cross-correlated chemical shift modulations, specifically target concerted changes in the isotropic chemical shifts of the two nuclei associated with spatial fluctuations. Field-dependence of the relaxation rates permits identification of the parameters defining the chemical exchange rate constant under the assumption of a two-site exchange. The time scale of motions falls into the intermediate to fast regime (with respect to the chemical shift time scale, 100–400 s^?1 range) for the ¹³C′–¹⁵N pairs and into the slow to intermediate regime for the ¹³C_α–¹³C_β pairs (about 150 s^?1). Comparison of the results obtained for protonated and deuterated GB3 suggests that deuteration has a tendency to reduce these slow scale correlated motions, especially for the ¹³C_α–¹³C_β pairs.     

Orientational interactions in low-concentration DNA solutions

The rotational relaxation tiem τ₃ of DNA molecules (M_w ? 5 × 10⁶) in solution has been determined by the transient electric birefringence method. The analysis of the birefringence decay makes it possible to study only the higher-molecular-weight fraction, the molecules being considered as rigid elongated particles in a short time scale. A marked concentration dependence of the relaxation time has been observed for DNA in low ionic strengths. Above a critical concentration c*, τ₃ increases with the DNA concentration, c. The value of c* increases with the ionic strength. For 10^?3 ionic strength (with NaCl), c* is about 10 μg/mL; then we observe the same strong concentration dependence of rotational relaxation times as recently reported for rodlike M-13 viruses [Maguire, J. F., McTague, J. P. & Rondelez, F. (1980) Phys. Rev. Lett. 45 , 1891–1894]. These results may be discussed in terms of the Doi-Edwards theory for rotational relaxation time of rigid macromolecules [Doi, M. (1975) J. Phys. 36 , 607–611; Doi, M. & Edwards, S. F. (1978) J. Chem. Soc. Faraday Trans. 74 , 918–932] and the critical concentration above which the interactions between the molecules begin to appear allows determining the corresponding molecular length. We observe a very good agreement between the DNA lengths obtained from the c* values and by using the infinite dilution value of τ₃ and Broersma's equation. Therefore, only highly diluted solutions can be used if intrinsic molecular properties based on the rotational diffusion of high-molecular-weight elongated molecules are studied.     

Effect of replacement of Ca by Zn on the structure and optical property of CaTiO3:Eu3+ red phosphor prepared by sol‐gel method

Eu³⁺‐doped calcium titanate red phosphors, Ca_1‐xZn_xTiO₃:Eu³⁺, were prepared by the sol‐gel method. The structure of prepared Ca_1‐xZn_xTiO₃:Eu³⁺ phosphors were investigated by X‐ray diffraction and infrared spectra. Due to the ⁵D₀ → ⁷F_1–3 electron transitions of Eu³⁺ ions, photoluminescence spectra showed a red emission at about 619 nm under excitation of 397 nm and 465 nm, respectively. When zinc was added to the host, the luminescent intensity of Ca_1‐xZn_xTiO₃:Eu³⁺ was markedly improved several fold compared with that of CaTiO₃:Eu³⁺. Ca_0.9Zn_0.1TiO₃:Eu³⁺ also had higher luminescence intensity than the commercially available Y₂O₃:Eu³⁺ phosphors under UV light excitation. Copyright © 2014 John Wiley & Sons, Ltd.     

Transient state isoelectric focusing. Experimental determination of apparent diffusion coefficients in polyacrylamide gels

A photoelectric scanning assembly utilizing uv absorption optics and an on-line digital data acquisition and processing system has been used to follow kinetically zone spreading during the defocusing stage (absence of electric field) of transient state isoelectric focusing (TRANSIF) in polyacrylamide gels. Measurement of the variance (σ²) of a diffusing zone as a function of time yields a linear relationship, the slope of which corresponds to the apparent diffusion coefficient (D) of the protein. A linear relationship is also obtained when the logarithm of the apparent diffusion coefficients (logD) are plotted vs acrylamide concentration (T). This relationship can be used to extrapolate D to zero gel concentration. The apparent diffusion coefficient measured in this way is significantly larger than the true diffusion coefficient. The slope of the plot logD vs T, designated C_R, is expected to be a measure of molecular size related to the retardation coefficient in polyacrylamide gel electrophoresis.