Dielectrophoresis of Cells

Dielectrophoresis, the motion produced by the action of nonuniform electric field upon a neutral object, is shown to be a simple and useful technique for the study of cellular organisms. In the present study of yeast (Saccharomyces cerevisiae) using a simple pin-pin electrode system of platinum and high-frequency alternating fields, one observes that the collectability of cells at the electrode tip, i.e. at the region of highest field strength, depends upon physical parameters such as field strength, field uniformity, frequency, cell concentration, suspension conductivity, and time of collection. The yield of cells collected is also observed to depend upon biological factors such as colony age, thermal treatment of the cells, and chemical poisons, but not upon irradiation with ultraviolet light. Several interesting side effect phenomena coincident with nonuniform electric field conditions were observed, including stirring (related to “jet” effects at localized electrode sites), discontinuous repulsions, and cellular rotation which was found to be frequency dependent.     

The continuous positive and negative dielectrophoresis of microorganisms

The continuous dielectrophoresis of living cells is described. The technique uses stream-centered transport of suspended microorganisms through an especially shaped non-uniform electric field. The cells can be given a positive or negative displacement, i.e., can be pushed into or out of the region of higher field intensity, depending upon the frequency of the applied ac field, and upon the relative permittivities of the cells and the suspending medium. Yeast (Saccharomyces cerevisiae) and algal cells (Chlorella vulgaris) were found to provide spectra of dielectrophoretic responses varying with the applied frequency (10 to 600 kHz) and conductivity.     

Receptor Autoradiography: The Last Ten Years

Abstract

The development of techniques for the microscopic visualization of receptor binding and other binding sites using autoradiography is reviewed. The application of these techniques to a number of pharmacological problems, particularly in the field of neuropharmacology is illustrated. The limitations and future trends in the field of receptor visualization are commented upon.     

Reflections provoked by Michael Banton's John Rex's main mistake

Banton's article provoked a number of reflections on working with John Rex in the 1960s. The idea of race has a long history both in human relations and sociological theory. But the possibility of there being a specific field of ‘race relations’ remains contested. Ideas of race and ethnicity are elided in public discourse, making the resolution of linguistic issues in the field especially difficult and as yet unresolved. These reflections touch upon the political environment in which John Rex's earlier writing on race was conducted and upon the breadth of influences upon a sociologist rooted in the classical tradition, especially the sociology of Max Weber.     

The Field of Art Production and Western Desert Acrylics1

Following the logic of Bourdieu's notion of ‘field’, acrylic art production in the Western Desert of Australia is positioned as a sub‐field in the field of Western art production. The significance of the cultural identity of the producer subordinates Western Desert acrylic art within the field and the shift in the criteria of legitimation constitutes it as a sub‐field. The capital at stake in the sub‐field of Western Desert acrylic art is differentiated and contingent upon legitimation of authenticity as defined by the Western field of art production. This capital is constituted through the positioning of Western Desert acrylics within the field of art production and is a manifestation of the struggle to maintain domination within the field.     

Highly sensitive field test lateral flow immunodiagnostics of PVX infection

A test system is described and expanded upon for mass field immunochromatography assay on porous membrane carriers for rapid diagnostics of potato virus X (PVX) in potato leaf tissue and sprout extracts using colloidal gold nanoparticles as a marker. Sensitivity of the assay developed for PVX identification is found to be comparable to the sensitivity of solid-phase sandwich-ELISA. Complete assay time does not exceed 15 min, and the lower limit of the PVX detection in non-clarified leaf extract is 2 ng/ml. A single measurement requires 0.1–0.2 ml (3–5 drops) of tested solution only (extracted from 10–20 mg of potato leaf tissue or sprouts). The simplicity and reliability of the method makes it especially efficient in direct rapid monitoring of many infected potato specimens in the field, as verified by field trials of 360 clones of 28 domestic and foreign cultivars of potato. A diagnostic kit for routine analyses of potato viral infections both in the laboratory and in the field is described and expanded upon.     

Modelling Beauveria bassiana horizontal transmission

1 We modelled horizontal transmission of Beauveria bassiana in Colorado potato beetle (CPB) between larval cadavers and soil inhabiting prepupae. 2 The rate of disease transmission, based on the probability of a prepupa contacting sporulating cadavers on the soil surface, is a non‐linear function of cadaver density and also dependent upon temperature. 3 The spatial pattern of cadavers is needed to estimate prepupal contacts with cadavers. Observational field studies determined Johnson distributions to model the spatial pattern of cadavers and prepupae in the field. The model also implicitly incorporates within‐field larval spatial pattern into estimates of field‐level horizontal transmission. 4 Potential for horizontal transmission is higher in simulations using weather data from the warmer year of 1995, than in simulations of the cooler growing season of 1993. 5 Simulations of CPB populations under northern Maine climatic conditions recorded in 1993 and 1995 suggest that horizontal transmission can range from 3 to 24% depending upon the timing of primary infection of larvae in the field. 6 Two simulated sequential applications of B. bassiana targeted at peak first instars resulted in maximum horizontal infection in both years. 7 Sensitivity analysis suggests that horizontal transmission is most sensitive to changes in the proportion of cadavers that sporulate and least sensitive to changes in the time between larval death and the onset of cadaver sporulation. 8 Field validation of the model indicates good prediction of one measure of horizontal infection, the proportion of prepupae which eventually sporulated after being released in controlled field experiments.     

Eucaryotic vs Procaryotic Metabolic Response to a Low-Amplitude Swept-Frequency Magnetic Field

Respiration of infant rat cells and of the intestinal flora of the adult rat was measured upon exposure to a swept-frequency (100–200 Hz) magnetic field of 5 mGauss, rms, to determine the metabolic effects of small changes in the ambient magnetic field upon these life forms. Eucaryotic metabolism was found to change sharply after the field was imposed and after it was eliminated, with a recovery both in and out of the imposed field. Procaryotic metabolism decreased gradually upon exposure, and exhibited no recovery. The results suggest a basic difference in the biological systems which may mark a crucial step in the development of eucaryotic biology.     

Structural transition produced by electric fields in aqueous sodium deoxyribonucleate

It was found that the birefringence of aqueous solutions of sodium DNA is anomalous when electric fields of high intensity (≥10⁴ v/cm) are applied. The magnitude of the birefringence first rose upon application of the orienting pulse, then fell as the field was sustained above a critical value. The occurrence of the effect depended upon macromolecular and electrolyte concentrations. Upon removal of the field, the birefringence was rapidly restored and then it decayed with an increase of the reorientational relaxation times, relative to those observed below the critical field. It is proposed that the electric field may cause aggregation of the macromolecules and then produce a structural transition concomitant with the electric field orientation effect. This transition may correspond to the “B” “A” structures identified in x-ray studies, or to a “B” “V” structure change, where “V” is a postulated new helical form stabilized by cooperative interactions of base and dipoles in the electric field. Field induced transitions of this type would be of interest in connection with molecular mechanisms of transport through membranes, nerve impulse transmission, or information storage.     

Electrorheological effects and gating of membrane channels

A hypothesis is presented on the gating of ion channels. This is considered as a consequence, in part, of a large increase in viscosity of the water in the "vestibule" region of the channel in the high field present when the channel is not conducting. This part of gating amounts to "melting" of the high viscosity part of the water upon release of the field. The resulting model accounts qualitatively for a number of phenomena in the literature, including the steepness of the voltage dependence of gating, the slowing of gating upon substitution of D2O for H2O, and the pressure dependence of the gating kinetics. The viscosity increase with field is well known in the literature; several forms of electroviscous effects, a viscoelectric effect, and a generalized electrorheological effect have been described. This model appears closest to an electrorheological effect in which boundary water out to a few molecular diameters is structured in the presence of a high field, while the boundary (here, protein) moves. The size of the channel entrance is small enough for this effect to prevent conductivity. The remainder of the gating current, which occurs at more polarized potentials, is attributed to protein motion. Some consequences of the model are discussed. Qualitative comparison with published data is included.     

Medium polarization effect on proton potential shape S.A SCRF-MO CNDO2 study of methanol and methanethiol H-bonded to imidazol

The theoretical calculations carried out with the self-consistent reaction field theory of solvent (medium) effects,have shown that the proton potential curve shapes of the above mentioned H-bonded systems are remarkably dependant upon the reaction field susceptibility parameter. The results show that the imidazol system would display different pK's at different sites of a given globular protein. Moreover, a proton translocation process is shown to be an essentially medium driven effect.     

Environmental effects on H-bond potentials: A SCRF MO CNDO/2 study of some model systems

The self-consistent reaction field (SCRF) theory of solvent effects upon dipolar species has been applied to the study of H-bonded systems. A model to represent the enzyme core medium is discussed. It is shown, albeit heuristically, that this type of environment may be represented with the combined effect of a reaction field and an inhomogeneous external electric field acting over the site system, this latter being provided by the main chain dipolar peptide residues. The generalization of the reaction field concept attained within the SCRF theory allowed for this extention to be operationally implemented.The model systems considered are: a formaldehyde-water, acetone-water, a water dimer and a tetrahedrally arranged water trimer. The interest has been focuzed on the study of the dependance upon the site-surrounding coupling strength of the intermolecular potentials and interactions related to the H-bond. The orientational and intermolecular distance dependance of the pair potential and the proton potentials have been considered.The perturbing effect of the enzyme core medium is manifested through the drastic variation of many (but not all) of the H-bond potentials. The changes showed by the orientational component of the intermolecular potential together with that of the proton potential shape illustrate this point. As far as the proton and charge relay mechanisms are concerned, the results reported, while confirming our previous ones, shed a new light upon these matters. The sensitization of the H-bridge towards an external electric field is made possible by the presence of the reaction field, in absence of this latter, the external field effect is not significant.     

A theory of charge separation,ion, electron and proton transport in photosynthetic membranes based on asymmetry of surface charges

We present a model for the light-induced charge separation, proton and ion transport across photosynthetic membranes based on an assumption of the transmembrane surface charge asymmetry. In dark equilibrium, this asymmetry gives rise to an internal membrane electric field whose direction is perpendicular to the membrane surfaces. The role of the field in the light-induced charge separation is similar to the function of the built-in electric field across a solid-state p-n junction. Light-generated free charge carriers in the membrane flow according to its direction and upon recombination on the surface give rise to an electrochemical potential difference for electrons across the membrane. The associated coupled electron-proton transport, and ion diffusion can be viewed as a response of the system to the light-induced redox and electric potential changes.     

Revision of the genus philactis (Compositae)

Philactis (Compositae-Heliantheae) is a genus of three species, all of limited distribution. A revision is presented, based upon herbarium specimens and field work. Cytological data are from bud material fixed in the field, and from root tips of seedlings. The basic chromosome number appears to be 14.P. nelsonii andP. liebmannii are tetraploid (2n = 56), andP. zinnioides is hexaploid (2n = 84).     

THE APPLIED BIOLOGIST, THE CHILD, AND SOCIETY

Consideration of the characteristics of past and current applied biology reveals its fundamentally ecological nature, and also indicates the potent influence of the applied biologist upon society. It is urged that in order to impress influence upon pupils biological education should be based mainly upon field studies, rather than upon predominantly evolutionary, morphological and physiological studies. The nature of past and existing school biology strongly reinforces the abovel conclusion.
The results of attempts to find and evaluate evidence of the social influences of past and present biological education are presented, both in relation to the whole population, as well as to particular aspects, like food poisoning, the press, and the education of Civil Servants and Members of Parliment.
The potentialities of a Sixth Form biology resting chiefly upon field study are discussed not only as a means of developing an outlook valuable to-day, but also of treating suitably the pre-university education of future applied biologists.     

Dielectrophoretic force: A comparison of theory and experiment

Because of the increasing use of dielectrophoresis in the dielectric characterization and sorting of living cells or their parts, it has become important to establish carefully the theoretical backgrounds for this effect. A comparison with experiment is made of the several versions of the theory for the dielectrophoretic force exerted by nonuniform electric fields upon a neutral object. The three fundamental approaches: the Maxwell-Strattonstress tensor, the effective dipole moment, and the ‘Helmholtz’ energy approach are presented along with the general solution given earlier by Pohl and Crane. These are found to agree closely with experiment in predicting the dielectrophoretic force upon various rods hung in specially shaped (isomotive) field distributions. On the other hand, an alternative formulation based upon a debatable assignment of fields local to the dipoles gave a good fit to the experimental data only for materials of very low permittivity, and fitted poorly in the case of highly polarizable materials. An improved derivation of the theory for stable dielectrophoretic levitation is also presented. This phenomenon is of particular interest in that it is based upon an apparent violation of the Earnshaw's theorem, and is useful in the study of the dielectric properties of individual living cells.     

Shoreline algal wash as a factor in reed decline in Lake Constance-Untersee

This paper assesses the chemical and mechanical impact of algal wash (Cladophora, Spirogyra, Chara) upon the lakeside reed belt (Phragmites australis) using field mapping methods, bioassays with Scenedesmus acutus in batch culture, and field experiments. Heavy mats of filamentous algae are correlated with a reduction in number of the outermost reed stalks. The water pressed from decaying heaps of Cladophora and Spirogyra reduced the growth rate of Scenedesmus significantly, but mats from Chara did not. It is assumed that the toxic substance is an organic compound. In field experiments the detrimental effect could not be clearly evidenced. The reasons for this are discussed. It is concluded that mechanical impact is of major importance.     

GROWTH AND DEVELOPMENT OF THE RETICULATE THALLUS IN THE LICHEN RAMALINA MENZIESII

The thallus of the lichen Ramalina menziesii Tayl. is comprised of subunits which resemble planar nets. The nets arise upon older nets as buds which develop perforations and expand. Microphotographic study of thallus development in the field shows several features which differ from a recently proposed model of development for this species. New perforations develop continually in new tissue produced at the apical margin of the net. The apical margin is inrolled, and may furcate. This type of branching is developmentally distinct from the formation of new nets from buds borne upon expanded parts of the net. Anatomical study of net apices appears to support assertions that programmed tissue necrosis contributes to perforation formation at sites where gaps are present in the algal layer.     

Polarization of fluorescence of an oriented solution of rodlike particles bearing a fluorescent label

The variation of the polarized components of fluorescence of a rodlike particle bearing a fluorescent label upon partial orientation is calculated for some special geometry of the dye macromolecules complexes. Explicit expressions are given for the case where the energy of the molecule in the field depends only on one angle θ, showing that the result is a function of both 〈sin²θ〉 and 〈sin⁴θ〉. For the case of orientation in an electric field through an anisotropic induced moment, the expressions allow the calculation of this anisotropy of polarizability. The method is applied to the measurement of the polarizability of rodlike fragments of DNA labeled by intercalated molecules of Acridine Orange.