Aminoalkyl adenylate and aminoacyl sulfamate intermediate analogues differing greatly in affinity for their cognate Staphylococcus aureus aminoacyl tRNA synthetases.

Aminoalkyl adenylates and aminoacyl sulfamates derived from arginine, histidine and threonine, have been prepared and tested as inhibitors of their cognate Staphylococcus aureus aminoacyl tRNA synthetases. The arginyl derivatives were both potent nanomolar inhibitors of the Class I arginyl tRNA synthetase whereas for the Class II histidyl and threonyl tRNA synthetases, the acyl sulfamates were potent inhibitors but the adenylates had very little affinity.

Mechanistic insights into the inhibition of serine proteases by monocyclic lactams.

Although originally discovered as inhibitors of pencillin-binding proteins, beta-lactams have more recently found utility as serine protease inhibitors. Indeed through their ability to react irreversibly with nucleophilic serine residues they have proved extraordinarily successful as enzyme inhibitors. Consequently there has been much speculation as to the reason for the general effectiveness of beta-lactams as antibacterials or inhibitors of hydrolytic enzymes. The interaction of analogous beta- and gamma-lactams with a serine protease was investigated. Three series of gamma-lactams based upon monocyclic beta-lactam inhibitors of elastase [Firestone, R. A. et al. (1990) Tetrahedron 46, 2255-2262.] but with an extra methylene group inserted between three of the bonds in the ring were synthesized. Their interaction with porcine pancreatic elastase and their efficacy as inhibitors were evaluated through the use of kinetic, NMR, mass spectrometric, and X-ray crystallographic analyses. The first series, with the methylene group inserted between C-3 and C-4 of the beta-lactam template, were readily hydrolyzed but were inactive or very weakly active as inhibitors. The second series, with the methylene group between C-4 and the nitrogen of the beta-lactam template, were inhibitory and reacted reversibly with PPE to form acyl-enzyme complexes, which were stable with respect to hydrolysis. The third series, with the methylene group inserted between C-2 and C-3, were not hydrolyzed and were not inhibitors consistent with lack of binding to PPE. Comparison of the crystal structure of the acyl-enzyme complex formed between PPE and a second series gamma-lactam and that formed between PPE and a peptide [Wilmouth, R. C., et al. (1997) Nat. Struct. Biol. 4, 456-462.] reveals why the complexes formed with this series were resistant to hydrolysis and suggests ways in which stable acyl-enzyme complexes might be obtained from monocyclic gamma-lactam-based inhibitors.

The accuracy of a new system for estimating organ volume using ultrasound.

A new system is described for estimating volume from a series of multiplanar 2D ultrasound images. Ultrasound images are captured using a personal computer video digitizing card and an electromagnetic localization system is used to record the pose of the ultrasound images. The accuracy of the system was assessed by scanning four groups of ten cadaveric kidneys on four different ultrasound machines. Scan image planes were oriented either radially, in parallel or slanted at 30 degrees to the vertical. The cross-sectional images of the kidneys were traced using a mouse and the outline points transformed to 3D space using the Fastrak position and orientation data. Points on adjacent region of interest outlines were connected to form a triangle mesh and the volume of the kidneys estimated using the ellipsoid, planimetry, tetrahedral and ray tracing methods. There was little difference between the results for the different scan techniques or volume estimation alogorithms, although, perhaps as expected, the ellipsoid results were the least precise. For radial scanning and ray tracing, the mean and standard deviation of the percentage errors for the four different machines were as follows: Hitachi EUB-240, -3.0 +/- 2.7%; Tosbee RM3, -0.1 +/- 2.3%; Hitachi EUB-415, 0.2 +/- 2.3%; Acuson, 2.7 +/- 2.3%.

Application of a new discreet form of Gauss' theorem for measuring volume.

Volume measurements are useful in many branches of science and medicine. They are usually accomplished by acquiring a sequence of cross sectional images through the object using an appropriate scanning modality, for example x-ray computed tomography (CT), magnetic resonance (MR) or ultrasound (US). In the cases of CT and MR, a dividing cubes algorithm can be used to describe the surface as a triangle mesh. However, such algorithms are not suitable for US data, especially when the image sequence is multiplanar (as it usually is). This problem may be overcome by manually tracing regions of interest (ROIs) on the registered multiplanar images and connecting the points into a triangular mesh. In this paper we describe and evaluate a new discreet form of Gauss' theorem which enables the calculation of the volume of any enclosed surface described by a triangular mesh. The volume is calculated by summing the vector product of the centroid, area and normal of each surface triangle. The algorithm was tested on computer-generated objects, US-scanned balloons, livers and kidneys and CT-scanned clay rocks. The results, expressed as the mean percentage difference +/- one standard deviation were 1.2 +/- 2.3, 5.5 +/- 4.7, 3.0 +/- 3.2 and -1.2 +/- 3.2% for balloons, livers, kidneys and rocks respectively. The results compare favourably with other volume estimation methods such as planimetry and tetrahedral decomposition.

Volume estimation from multiplanar 2D ultrasound images using a remote electromagnetic position and orientation sensor.

A system is described for calculating volume from a sequence of multiplanar 2D ultrasound images. Ultrasound images are captured using a video digitising card (Hauppauge Win/TV card) installed in a personal computer, and regions of interest transformed into 3D space using position and orientation data obtained from an electromagnetic device (Polhemus, Fastrak). The accuracy of the system was assessed by scanning 10 water filled balloons (13-141 mL), 10 kidneys (147-200 mL) and 16 fetal livers (8-37 mL) in water using an Acuson 128XP/10 (5 MHz curvilinear probe). Volume was calculated using the ellipsoid, planimetry, tetrahedral and ray tracing methods and compared with the actual volume measured by weighing (balloons) and water displacement (kidneys and livers). The mean percentage error for the ray tracing method was 0.9 +/- 2.4%, 2.7 +/- 2.3%, 6.6 +/- 5.4% for balloons, kidneys and livers, respectively. So far the system has been used clinically to scan fetal livers and lungs, neonate brain ventricles and adult prostate glands.

Dielectric properties of human blood at microwave frequencies.

Waveguides systems have been used to obtain measurements for the complex permittivity of human blood in the frequent range 29-90 GHz. The data are reported and analysed in conjunction with lower frequency work, in particular that of Wei. There are two distinct regimes of behaviour in the spectra obtained: a high-frequency relaxation process in parallel with a lower frequency fractional power-law response.

The dielectric properties of rabbit tissue, pure water and various liquids suitable for tissue phantoms at 35 GHz.

Newly developed microwave apparatus was used to measure the complex permittivity of some rabbit tissues and water over the temperature range 20-37 degrees C. Various liquid mixtures which might be suitable as phantom tissue equivalents at 35 GHz were also considered. Some of the measured tissue data were compared with previous lower frequency data and it was concluded that the observed behaviour was not anomalous.

The modelling of biological systems in three dimensions using the time domain finite-difference method: I. The implementation of the model.

A computer method has been developed which uses the time domain finite-difference (TDFD) algorithm to calculate the deposition of the electromagnetic (EM) field in three-dimensional biological models. This, the first of two papers, describes the algorithm and the computer programs developed. The method is demonstrated by calculating the penetration of the EM field from a rectangular waveguide radiating into a homogeneous model, the calculation being carried out in two dimensions for simplicity in this paper.

The modelling of biological systems in three dimensions using the time domain finite-difference method: II. The application and experimental evaluation of the method in hyperthermia applicator design.

The three-dimensional time domain finite-difference (TDFD) method was used to design a 433 MHz circular waveguide applicator for electromagnetic hyperthermia. This applicator has a water-filled lens at one end, providing surface cooling as well as some shaping of the emergent electromagnetic field. An experimental system for scanning the EM field in three dimensions is described. The experimental data were compared with those calculated from the TDFD program and good agreement was found. A realistic three-dimensional model based on an ultrasound scan of a surface tumour was built. Both the EM field and the specific absorption rate (SAR) within the model when irradiated by the applicator were calculated.

Dielectric behavior of water in biological solutions: studies on myoglobin, human low-density lipoprotein, and polyvinylpyrrolidone.

The dielectric behavior of the aqueous solutions of three widely differing macromolecules has been investigated: myoglobin, polyvinylpyrrolidone (PVP), and human serum low-density lipoprotein (LDL). It was not possible to interpret unambiguously the dielectric properties of the PVP solution in terms of water structure. The best interpretation of the dielectric data on the myoglobin and LDL solutions was that, in both cases, the macromolecule attracts a layer of water of hydration one or two water molecules in width. For LDL, this corresponds to a hydration factor of only 0.05 g/g, whereas for myoglobin the figure is nearer 0.6 g/g. With myoglobin, part of the water of hydration exhibits its dispersion at frequencies of a few GHz, and the rest disperses at lower frequencies, perhaps as low as 10-12 MHz. The approximate constancy of the width of the hydration shell for two molecules as dissimilar in size as LDL and myoglobin confirms that the proportion of water existing as water of hydration in a biological solution depends critically on the size of the macromolecules as well as on their concentration.

Dielectric properties of lens tissue at microwave frequencies.

The relative permittivity and conductivity of rabbit eye lens were measured in the frequency domain between 2 and 18 GHz at temperatures of 37 and 20 degrees C. An analysis of the data suggested that a significant proportion of the bulk water in nuclear and cortical lens tissue may behave differently to pure water. In addition, the Maxwell-Fricke mixture theory was used to estimate the amount of hydrated water that relaxes far below 1 GHz.

Dielectric properties of mammalian brain tissue between 1 and 18 GHz.

A newly developed frequency domain technique was used to measure the dielectric properties of white matter, grey matter and macerated rabbit brain at 20 and 37 degrees C. An analysis of the data showed that between 1 and 4 GHz there were substantial contributions from processes other than the gamma dispersion. However, above 7.5 GHz it appeared that mainly free water was relaxing although evidence of a small spread of relaxation times was found for the bulk water in the white matter. Mouse and rat brain were also measured but no significant differences were found between the species. The quantity of bound water was estimated but there was no evidence of a difference in the amount between either the tissues or the temperatures.

Dielectric properties of developing rabbit brain at 37 degrees C.

The dielectric properties of developing rabbit brain were measured at 37 degrees C between 10 MHz and 18 GHz using time domain and frequency domain systems. The results show a variation with age of the dielectric properties of brain. An analysis of the data suggests that the water dispersion in the brain of newly born animals can be represented by a Debye equation. This dispersion increases in complexity with age, and there is evidence of a smaller additional relaxation process centered around 1 GHz. It is concluded that the principal contribution to this subsidiary dispersion region arises from water of hydration.

Microwave dielectric measurements (0.8-70 GHz) on Artemia cysts at variable water content.

Dielectric permittivity measurements are reported for cysts of Artemia, a crustacean known as the brine shrimp. Using coaxial and waveguide techniques we examined the frequency range from 0.8 to 70 GHz. Taking advantage of the ability of this system to reversibly lose essentially all intracellular water, we determined the permittivity over the entire range of cyst water contents. Although experimental errors prevent a rigorous treatment of the data, we advance the general conclusion that little of the water in this system behaves dielectrically like pure water, regardless of water content. This conclusion is supported by, and is consistent with, the results of previously published studies that probe the motional properties of water in this system using nuclear magnetic resonance spectroscopy and quasi-elastic neutron scattering.

Dielectric behavior of DNA solution at radio and microwave frequencies (at 20 degrees C).

The dielectric constant and conductivity of calf thymus DNA were investigated at frequencies between 0.1 MHz and 70 GHz. This work is to investigate the dielectric properties of DNA in low gigahertz region and also to study whether the dielectric behavior of the water is affected by the presence of highly charged DNA. The results of these measurements indicate the presence of two anomalous dispersions, the one between 1 MHz and 1 GHz and the second one above 1 GHZ. The dispersion at low frequencies is likely to arise from polar groups in the DNA molecule. The relaxation behavior of unbound water in DNA solution is only slightly affected by the presence of DNA at concentrations below 1%.

Dielectric properties of insulin in solution.

Values of the relative permittivity of a 5% solution of despentapeptide insulin (DPI) are given at six temperatures over a frequency range 0.2-50 MHz. The solvent was a mixture of water and ethylene glycol at pH = 3. From the dispersion curves a dipole moment of 72 +/- 1 was obtained for the insulin molecule at 25 degrees C. By combining the dielectric measurements with values of solvent viscosity the volume of the insulin molecule in solution was found to be only slightly larger than that in the crystal. This indicates that insulin possesses a markedly lower quantity of water of hydration than has been observed for globular proteins containing a higher proportion of hydrophilic groups on the surface of the molecule.

The dielectric properties of the cerebellum, cerebrum and brain stem of mouse brain at radiowave and microwave frequencies.

The relative permittivity and conductivity of the cerebellum, cerebrum and brain stem of mouse brain were measured at a temperature of 37 degrees C over a frequency range of 72 MHz to over 5 GHz using time-domain spectroscopy. An analysis of the data suggests that the water exists in various forms of binding with an average relaxation frequency less than free water.

The dielectric properties of normal and tumour mouse tissue between 50 MHz and 10 GHz.

Microwaves are currently being investigated as a method of inducing hyperthermia. Values of the relative permittivity and conductivity of mouse muscle and tumour tissues over the frequency range 50 MHz to 10 GHz at a temperature of 37 degrees C are reported. The data were obtained by using an on-line computer-based time-domain spectrometer. The tumour tissues were the RIF/1 and KHT fibrosarcomas sited in the hind leg muscle of C3H mice. A significant difference in the relative permittivity between the normal and tumour tissue was shown to exist at frequencies up to 200 MHz.

Dielectric properties of ocular tissues at 37 degrees C.

Values of the relative permittivity, dielectric loss and conductivity are given for various rabbit ocular tissues at frequencies in the range 10 MHz-10 GHz. The tissues measured were cornea, retina, choroid, iris, and the cortical and nuclear zones of the lens. The dielectric parameters were determined using the technique of multiple response time domain spectroscopy. For all tissues the water relaxation could be characterised by a Debye dispersion with a relaxation time longer than that of pure water, indicating that its dielectric behaviour was affected by the presence of the biological macromolecules.