Tadeusz Walczak

Kinetics of chromium(V) formation and reduction in fronds of the duckweed Spirodela polyrhiza — a low frequency EPR study

The uptake of chromate by the duckweed Spirodela polyrhiza was investigated with atomic absorption spectroscopy and the reduction of Cr(VI) to Cr(V) was measured using low frequency EPR spectroscopy. The biphasic kinetics of the uptake was fitted to parameters of a proposed kinetic model. Another model was developed to simulate chromate reduction. The first step of chromate reduction was found to be much faster than the uptake of Cr(VI) from the free space. Most probably, this step occurs already in the cell wall or on the cell membrane surface. Further reduction of Cr(V) to Cr(III) was estimated to be slower. The disappearance of the Cr(V) signal, following transfer of the plants into a Cr-free solution, lasted several tens of hours; the kinetics was mono- or biexponential depending on the length of Cr loading. The rate constants for Cr reduction in living plants were determined for the first time.

DEVELOPING IN VIVO EPR OXIMETRY FOR CLINICAL USE

This paper describes the rationale for carrying out electron paramagnetic resonance (EPR) oximetry studies in human subjects in the clinical setting and the potential approaches and specific steps needed to make such studies feasible and useful. While the approach is described specifically for EPR oximetry, many of the principles may apply to the initial clinical uses of other techniques. The suggested operational approach is to have the initial applications occur in as clinically useful and simple a manner as possible, with the expectation that once the technique is introduced and accepted in the clinical setting, that more complex and/or more technically difficult applications will be able to be developed. The initial approach will be based on EPR spectroscopy at 1.2 GHz focusing on applications for which in vivo EPR provides a clearly useful approach to important clinical problems for which currently there is no good alternative method. The EPR measurements can be carried out non-invasively by measurements within 10 mm of the surface after the placement of the paramagnetic material at the site of interest, or by the placement of a needle/catheter in the site of interest for the required time period. The suggested initial clinical applications are guiding therapy for individual patients with tumors or vascular disease, by direct measurements of tissue pO2.

In vivo EPR dosimetry of accidental exposures to radiation: experimental results indicating the feasibility of practical use in human subjects

Low frequency electron paramagnetic resonance (EPR) provides the potential advantage of making accurate and sensitive measurements of absorbed radiation dose in teeth in situ, i.e. without removing the teeth from the potential victim. The potential limiting factors for making such measurements are: (1) whether low frequency EPR is sufficiently sensitive to detect radiation-induced signal in human teeth; (2) whether sufficient sensitivity can be maintained under in vivo conditions. In this manuscript, we summarize results indicating that this approach is feasible. Using 1.2 GHz EPR spectroscopy, we found that the lower limit for these measurements in isolated human teeth is 0.2 Gy or lower. Measurements of radiation-induced EPR signals in the teeth of living rats were achieved with sufficient sensitivity to indicate that, when taking into consideration the larger mass of human teeth, similar measurements in human teeth in situ would provide sensitivity in the dose range for potential accidental exposures. We estimate that the current lower limit for detecting radiation doses in human teeth in situ (in vivo) is 0.5-1.0 Gy; this would be sufficient for determining if a person has been exposed to potentially life threatening doses of ionizing radiation. The limiting factor for sensitivity appears to be background signals rather than signal/noise, and there are feasible means to overcome this problem and further increase sensitivity. The additional instrumental developments required to make an effective in vivo EPR dosimetric spectrometer for the measurements in teeth in human subjects in situ, seem quite achievable.

EPR spectrometer for clinical applications

This article describes an EPR spectrometer specifically designed and constructed for EPR spectroscopy in humans. The spectrometer is based on a permanent magnet, suitable for measurements at 1200 MHz. The magnet has a full 50 cm gap between the poles, which facilitates accurate and comfortable placement of the subject for the EPR measurement at any location on the human body. The bridge includes features to facilitate clinical operations, including an indicator for phasing of the reference arm and a 2 level RF amplifier. Resonators with holders for each type and site of measurement have been developed that comfortably position the resonator and the patient and prevent artifacts due to motion. The initial applications for which the spectrometer has been designed are for oximetry using loops on the surface, oximetry using implanted resonators for measuring deep sites, and measurements in the teeth for determination of exposures to clinically significant doses of ionizing radiation. Magn Reson Med, 2005.

Blue-light-induced chloroplast orientation in Mougeotia. Evidence for a separate sensor pigment besides phytochrome.

Chloroplast orientation in the green alga Mougeotia has been induced by unidirectional red or blue light, given continuously during one hour. In addition, part of the preparations obtained scattered strong far-red light simultaneously with the orienting light. This far-red light completely abolished the response to red light, consistent with phytochrome as the sensor pigment for orientation in Mougeotia. In blue light, however, the response was completely insensitive to far-red light, thus pointing to a different sensor pigment in the shortwavelength region.

An improved external loop resonator for in vivo L-band EPR spectroscopy.

An improved external loop resonator (ELR) used for L-band electron paramagnetic resonance (EPR) spectroscopy is reported. This improvement is achieved by shortening the parallel coaxial line. The resonant structure is formed by two single turn coils (10mm in diameter) that are connected to a parallel coaxial line. A resonance frequency of 1197 MHz and a quality factor of 466 were obtained in the absence of biological tissue and were approximately 1130 MHz and approximately 50 with a living animal, respectively. The sensitivity of the new ELR was compared to the previously developed ELR using three types of EPR samples: (1) paramagnetic material with no biological tissue, (2) paramagnetic material in a leg and in the peritoneal cavity of a dead rat, and (3) paramagnetic material in the back of an anesthetized rat. The sensitivity was 1.2-1.6 times greater in the rat and 4.2 times without tissue.

Measurements of pO2 in Vivo, Including Human Subjects, by Electron Paramagnetic Resonance

The purpose of this paper is to provide an illustrative description of the current state of development of the use of electron paramagnetic resonance (EPR, or completely equivalently, electron spin resonance or ESR) to measure the partial pressure of oxygen (pO2) in tissues in vivo under physiological conditions. This summary is based on published and unpublished results from our laboratory (1–7) and does not attempt to describe the results of other laboratories which also are working along related lines (8–10). The pertinent features of our technique are illustrated. We also consider the current limitations of the technique and likely developments in the near future. Our evaluation is that: this technique now is suitable for immediate use in small animals; within a short period of time instruments will be available facilitating its use in larger animals; and preliminary studies are imminent in human subjects (7).

INTERACTION BETWEEN PHYTOCHROME AND THE BLUE LIGHT PHOTORECEPTOR SYSTEM IN Mougeotia

Abstract— Face‐to‐profile chloroplast movement in Mougeotia was induced by sequences of strong blue and red short irradiations. This type of response occured only when blue light was applied prior to or simultaneously with red light, and far‐red irradiation was necessary after the sequence to cancel the remaining gradient of the far‐red absorbing form of phytochrome Pfr. The dependence of the response magnitude on blue and red light sequences was studied for a wide range of light durations and dark intervals. The relationship between the response and the dark interval points to the lack of direct coupling between phytochrome and blue‐absorbing “cryptochrome”. It was postulated that a photoproduct having a life‐time of2–3 min is formed by the blue‐light‐mediated reaction. This photoproduct interacts with phytochrome during its transformation or with its final Pfr form.

Characteristics of an electronically tunable surface-coil-type resonator for L-band electron paramagnetic resonance spectroscopy

This note describes considerable additional information on the key parameters of an electronically tunable surface-coil-type resonator for L-band continuous-wave electron paramagnetic resonance (EPR) spectroscopy [Hirata et al., J. Magn. Reson. 142, 159 (2000)]. For the 1.1 GHz prototype tunable surface coil (10.6 mm in diameter), the measured efficiency for generating the rf magnetic field is 77 μT/W1/2. The unloaded quality factor of the tunable surface coil is in the range of 260–310. In absence of an EPR sample, 57% of the magnetic energy in the system is stored in the parallel coaxial line, 16% in the surface coil, and a half of the energy is dissipated in the parallel coaxial line. The energy dissipation for the surface coil occurs mainly through radiation (25% of the total energy dissipation). The level of the second harmonic in the reflected microwaves generated by the tunable surface coil was measured. The varactor diodes used in the matching and tuning circuits are the dominant source of the sec...

L-band electron paramagnetic resonance spectrometer for use in vivo and in studies of aqueous biological samples

The development of L-band (∼1.2GHz) frequency EPR spectrometers has made feasible many in vivo studies in laboratory animals and, recently, in human volunteers. The lower dielectric and eddy current losses that occur at L-band balance the lower Zeeman splitting so useful measurements can be made in conductive aqueous samples. We describe typical resonators used in such studies and provide details on the construction of the spectrometer, including the bridge, the automatic frequency control subsystem, the low-noise high-stability tunable L-band frequency source, as well as the low-frequency components—the signal receiver and the modulation unit. The application of EPR spectroscopy to larger subjects requires special care in the design of an appropriate magnet with sufficient homogeneity and stability, yet with dimensions that allow operation with a wide range of subject sizes. We describe our solution, which involves a permanent magnet, air-core scan coils to provide the field sweep and offset, and field s...