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U Pliquett

Publications and source records attributed to U Pliquett.

15 recordsLinked to original sources
PubMed2005

Local and transient structural changes in stratum corneum at high electric fields: contribution of Joule heating.

Electroporation of skin is accompanied by local heating, such that thermally induced structural changes of the stratum corneum (SC) accompany the field effect. Comparing on the time scale, the local changes in structure, temperature and conductance of the SC, during and after the pulse, it is seen that Joule heating also facilitates the subsequent molecular transport. It is found that the transport of medium-sized, ionic molecules occurs through localized transport regions (LTR). The size of a LTR increases with the pulse length, whereas the density of the LTRs increases with increasing voltage, for instance at U(SC=)80 V, the LTR cover approximately 0.02--1% of the surface area. The state of low resistance within the LTR is long-lived. During high voltage application, the center of the LTR is heated above the phase transition temperature of the SC lipids (70 degrees C) and the heat front propagates outwards. Inside the SC, the pulse causes aggregates of small-sized vesicles. At a higher temperature, the aggregate formation and their disappearance are delayed. Multiple pulses with the applied voltage of U(appl)=80 V induce the formation of long-lasting vesicle aggregates with a diameter of slashed circle=1--30 microm, covering 0.05--0.5% of the total sample area. The electric energy dissipated within the LTR during high voltage application is apparently sufficient to raise the temperature well above the phase transition temperature of the lipids of the SC, accounting for the conformational changes from the multi-lamella to the vesicular structures.

Animals↗
PubMed2004

Electroporation of subcutaneous mouse tumors by rectangular and trapezium high voltage pulses.

The artificial electrotransfer of bioactive agents such as drugs, peptides or therapeutical nucleic acids and oligonucleotides by membrane electroporation (MEP) into single cells and tissue cells requires knowledge of the optimum ranges of the voltage, pulse duration and frequency of the applied pulses. For clinical use, the classical electroporators appear to necessitate some tissue specific presetting of the pulse parameters at the high voltage generator, before the actual therapeutic pulsing is applied. The optimum pulse parameters may be derived from the kinetic normal mode analysis of the current relaxations due to a voltage step (rectangular pulse). Here, the novel method of trapezium test pulses is proposed to rapidly assess the current (I)/voltage (U) characteristics (IUC). The analysis yields practical values for the voltage U(app) between a given electrode distance and pulse duration t(E) of rectangular high voltage (HV) pulses, to be preset for an effective in vivo electroporation of mouse subcutaneous tumors, clamped between two planar plate electrodes of stainless steel. The IUC of the trapezium pulse compares well with the IUC of rectangular pulses of increasing amplitudes. The trapezium pulse phase (s) of constant voltage and 3 ms duration, following the rising ramp phase (r), yields a current relaxation which is similar to the current relaxation during a rectangular pulse of similar duration. The fit of the current relaxation of the trapezium phase (s) to an exponential function and the IUC can be used to estimate the maximum current at a given voltage. The IUC of the falling edge (phase f) of the trapezium pulse serves to estimate the minimum voltage for the exploration of the long-lived electroporation membrane states with consecutive low-voltage (LV) pulses of longer duration, to eventually enhance electrophoretic uptake of ionic substances, initiated by the preceding HV pulses.

Animals↗
PubMed2004

Surface area involved in transdermal transport of charged species due to skin electroporation.

The electroporative effect on the stratum corneum (SC) is highly localized. However, the fractional area for the transport of small ions and larger ionic species differs considerably during and after high voltage (HV) application. Electroporation of SC creates new aqueous pathways, accessible for small ions, such as Cl(-) and Na(+) ions. The pores are distributed across the skin surface yielding a fractional area for current flow during electroporation of up to 0.1%. An increased permeability after high voltage application persists within a fractional area on the order of 10(-3)%. The permeabilization of SC for larger, charged molecules (M > 200 g/mol) involves Joule heating and a phase transition of the long chain sphingolipids within local transport regions (LTR). The transport area for these molecules (approximately 10(-3)%) changes only negligibly after high voltage application.

Biological Transport↗
PubMed2004

Prediction of lamb carcass composition by impedance spectroscopy.

The objective of this study was to compare impedance spectroscopy with resistance measurements at a single frequency (50 kHz) for the prediction of lamb carcass composition. The impedance spectrum is usually recorded by measuring the complex impedance at various frequencies (frequency domain); however, in this study, we also applied the faster and simpler measurement in the time domain (application of a current step and measurement of the voltage response). The study was carried out on 24 male, German Black-headed Mutton lambs with an average BW of 45 kg. Frequency- and time domain-based impedance measurements were collected at 20 min and 24 h postmortem with different electrode placements. Real and imaginary parts at various frequencies were calculated from the locus diagram. Left sides were dissected into lean, fat, and bone, and right sides were ground to determine actual carcass composition. Crude fat, crude protein, and moisture were chemically analyzed on ground samples. Frequency- and time domain-based measurements did not provide the same absolute impedance values; however, the high correlations (P < 0.001) between these methods for the "real parts" showed that they ranked individuals in the same order. Most of the time domain data correlated higher to carcass composition than did the frequency domain data. The real parts of impedance showed correlations between -0.37 (P > 0.05) and -0.74 (P < 0.001) to water, crude fat, lean, and fatty tissue, whereas the relations to CP were much lower (from 0.00 to -0.47, P < 0.05). Electrode placements at different locations did not substantially improve the correlations with carcass composition. The "imaginary parts" of impedance were not suitable for the prediction of carcass composition. The highest accuracy (R2 = 0.66) was reached for the estimation of crude fat percentage by a regression equation with the time domain-based impedance measured at 24 h postmortem. Furthermore, there was not a clear superiority of measurements in a wide frequency range over a single frequency measurement at 50 kHz for the prediction of carcass composition. Even though we calculated the impedance at 50 kHz based on the locus diagram, which allowed for a high precision for predicting this impedance trait, single-frequency impedance devices typically used in practice cannot record the locus diagram and, therefore, exhibit a greater amount of uncertainty.

Animals↗
PubMed2003

Joule heating during solid tissue electroporation.

The application of high-voltage pulses to biological tissue causes not only electroporation, a non-thermal phenomenon of pore creation within a lipid membrane due to an elevated electric field, but also significant heating. Once a biological membrane is porated, the current density increases several times, causing Joule heating. A combined experimental and theoretical study is reported. The theoretical temperature rise for a 1.25 kV cm(-1), 6 ms pulse is about 11.2 K for a tissue conductivity of 0.5 S m(-1) (i.e. myocardial tissue) during high-voltage application. Owing to the inhomogeneous electric field obtained with the use of needle electrodes, the temperature rises first at the electrodes, where the field strength reaches a maximum. Only for highly conductive tissue such as muscle was a temperature effect primarily observed in the bulk. Even if the temperature effect is biologically insignificant, it can affect the creation of stabile aqueous pathways by electroporation. The calculation of temperature distribution during high-voltage application, taking the electric field strength and the heat transfer into account, can be a useful tool for electrode optimisation.

Animals↗
PubMed1999

Changes in skin structure and electrical properties following high voltage exposure.

Human skin experiences extensive alterations when exposed to a strong electric field. In high voltage electric shocks, tissues in the current path undergo large-scale changes; the stratum corneum (SC), the outermost layer of the skin, loses its barrier function against ionic transport. The primary mechanism of electric field interaction with the skin is the creation of aqueous pathways, which increase the permeability of the skin to ions and macromolecules. Changes in skin structure and electrical properties were characterized by measuring the transport of fluorescent molecules in an in vitro preparation of human cadaver skin in a permeation chamber. Application of high voltage pulses (Uskia > 150 V, tau pulse = 1 ms) decreased the skin resistance per unit area by three orders of magnitude from its initial value of 100 k omega-cm-2. The resistance reached a quasi-steady state after about 20 pulses. After the cessation of pulsing, Rskin recovered immediately to about 4 k omega-cm-2 and remained low for 2 h thereafter. Transport of fluorescent molecules by high voltage pulses occurs in highly localized transport regions (LTRs), with diameters not exceeding 100 microns for short pulses (tau pulse < 5 ms).

Electric Impedance↗
PubMed1999

Dynamics of membrane sealing in transient electropermeabilization of skeletal muscle membranes.

Large supraphysiologic transmembrane electrical potentials are known to alter the molecular organization of the bilayer lipid component of cell membranes, leading to ionic permeabilization or "electroporation". Typically, membrane electroporation is followed by several orders of magnitude increases in electrical conductance and diffusive permeability to low-molecular-weight solutes. Electroporation may be transient or stable depending on whether the membrane eventually seals or remains permeabilized. Factors that control sealing have not been well characterized. This paper describes the kinetics of membrane sealing following electroporation by pulses over a range of supraphysiologic potentials. The increase in membrane conductance is highly nonlinear during a -440-mV, 4-ms pulse and reaches two orders of magnitude greater than baseline. Electroporation and relaxation sealing kinetics are quite different, reflecting a significant hysteresis effect. Thus, it appears that the magnitude and duration of the field pulse are important factors in sealing.

Animals↗
PubMed1999

Stress action on biological tissue and tissue models detected by the Py value.

The Py value, a fast measurable combination of the conductivity at the corner frequencies of the beta-dispersion, is a measure of the relative cell volume concentration in tissue. In many cases, if the biological object is stressed, for instance, by mechanical deformation, shortage of oxygen, electric field strength, temperature rise, or ischemia, Py increases. Depending on the object and the kind of stress, Py plateaus for minutes up to hours and then it decreases continuously. Values of passive electrical parameters of biological tissues are often given without information about the time following a stimulation or stress situation, for example, death, surgery, field application, etc. However, since the passive electrical properties change with time, information about their history, for example, time after death, should be given.

Electric Impedance↗
PubMed1999

Mechanistic studies of molecular transdermal transport due to skin electroporation.

The application of electrical high voltage pulses has been shown to greatly enhance the transdermal transport of water-soluble compounds. The resistance of the skins most important barrier, the stratum corneum, drops within less than 1 µs by orders of magnitude. This effect is attributed to electroporation, a nonthermic phenomena known to occur in phospholipid double layers. The striking difference between the stratum corneum lipid layers and the usually investigated phospholipid systems is the phase transition temperature. While lipid layers used for electroporation experiments are in liquid crystal phase above the phase transition temperature, the stratum corneum lipids (phase transition at approximately 70 degrees C) form a rigid quasi-crystalline membrane at room temperature.After the electrical stimulus a recovery of the passive flux was found making high voltage pulsing a suitable tool for controlling transdermal drug delivery. By ordinary light microscopy no dramatic changes in skin structure were found supporting the thesis of electroporation. However the microstructure shows clearly persistent structural changes. Recently the involvement of Joule heating due to the electric stimulus was shown as an important factor for skin permeabilization and molecular transport.

Journal Article↗
PubMed1996

Passive electrical properties of human stratum corneum in vitro depending on time after separation.

The passive electrical properties of human skin after separation from the body are predominated by the stratum corneum. Skin within a bath medium (150 mM phosphate buffered saline) at constant temperature (37 degrees C) exhibits a characteristic change of the passive electrical properties with time. Independent of the time the locus in the Z-plane is a depressed semicircular arc. The angle between the lines from the center of the arc to the points where the locus reaches the real axis remains unchanged. The difference between the high and low frequency resistivity (R0-Rx) increases over 10 h, reaches a plateau and decays after 20 h exponentially with a time constant of about 40 h. As model for the impedance we used a 5 element electrical circuit (R0, R1, R2, C1, C2), describing 3 pathways, (0) the dc path (appendages; R0), (1) tortuous pathways around the cell structures (R1, C1) and (2) direct pathways involving the corneocytes (R2, C2). There are characteristic changes with time in the elements of the equivalent circuit up to about 200 h after excision. Dramatic changes in C1 and R2 at about this time after separation strongly suggests destruction of the lipid structures. It will be suggested that the use of separated human stratum corneum as model for in vivo yields unreliable results after this time.

Electric Impedance↗
PubMed1995

Changes in the passive electrical properties of human stratum corneum due to electroporation.

The stratum corneum (SC) is the main barrier to molecular and ionic transport across mammalian skin and has been extensively studied by others at low voltages (U(skin)(t) < 10 V) in order to partially characterize the skin. Here we use one or more exponential pulses (tau pulse = 1 ms) and a temperature of 25 +/- 2 degrees C and found that the low voltage passive electrical properties (impedance) change rapidly and significantly if these pulse result in U(skin),0 > 40 V. In contrast, the dynamic resistance (describing passive electrical behavior in a nonlinear range) changes dramatically by application of pulses between 40 V and 80 V and then it settles at levels between 50 omega and 100 omega. We also found that recovery of the low voltage electrical parameters after pulsing depends mainly on the voltage, and, for multiple pulse protocols, on the number of pulses. For single pulses of U(skin),0 approximately 90 V or less the electrical recovery was almost complete, returning to within 0.90 of the pre-pulse value. In contrast, larger pulses result progressively in decreased recovery. The recovery for pulses > 90 V revealed several characteristic times, suggesting the involvement of different processes. For multiple pulses with U(skin),0 > 130 V almost no recovery of the transdermal resistance, R(skin), was evident (returning to < 0.10 of pre-pulse values), i.e., essentially permanent changes in the stratum corneum occurred. This is similar to that of single bilayer membrane electroporation, for which a transition from reversible to irreversible behavior occurs as transmembrane voltage is increased. Thus, these results are consistent with the hypothesis that 'high-voltage' pulses cause electroporation within the SC, i.e., that elevated transmembrane voltage result in creation of new aqueous pathways ('pores') across SC lipid regions.

Electric Conductivity↗
PubMed1995

Measurement of rapid release kinetics for drug delivery.

A fluorescence measurement system and methods of data analysis were developed to measure rapid kinetics of transdermal transport in vitro. Three variations on the technique were demonstrated, where the receptor compartment concentration was determined by: 1) fluorescence measurements of aliquots removed at discrete time points, 2) continuous fluorescence measurements made directly in the receptor compartment using a custom-made fluorimeter cuvette as a permeation chamber, and 3) continuous fluorescence measurements made in a flow-through cuvette containing receptor solution continuously pumped from a flow-through permeation chamber. In each case, the measured signal was a convolution of the time-dependent molecular flux (the desired information) and the characteristic response of the measurement system. Algorithms for deconvolution of the signal were derived theoretically. For the most complicated case, (3), the experimental confirmation is shown here, proving a time resolution on the order of half a minute.

Animals↗
PubMedDate not supplied

Evaluation of fast time-domain based impedance measurements on biological tissue.

Bio-impedance measurements are widely used for characterization of biological objects. Although the measured impedance of such objects is independent of the measurement method used, slight differences between measurements in the frequency and time domain are found. For many practical applications time domain based measurements are advantageous, but they are often rejected as not accurate. In order to show their suitability for bio-impedance measurements we used a special arrangement of time domain and frequency domain based measurements at the same biological specimen (canine liver) with the same electrodes. A reasonable coincidence in the measurement results could be shown. Moreover we used only a fraction of the time domain measurement data in order to demonstrate a significant reduction in measurement time while maintaining a reasonable accuracy. An algorithm for fast processing of the time domain data without transformation into the frequency domain is provided.

Animals↗