Localization of G-proteins in macrophages and E. coli during phagocytosis

Heterotrimeric GTP-binding proteins (G-proteins) have been shown to play an important role in cellular signalling. However, G-protein involvement in the intracellular spreading of bacterial pathogens is still poorly understood. In this study, antibodies, that recognize G-protein alpha-subunits (anti-G alpha), were used to investigate the localization of G-proteins in the macrophage-like cell line P388D1 and E. coli, also in their L-forms, during phagocytosis. In E. coli, anti-G alpha-binding sites were detected preferably in the cell wall and septa of the whole bacterial forms as well as in the cytoplasm of L-forms. Western blotting of bacterial lysates demonstrated protein bands with positive immunoreaction to antibodies against Gs alpha, Gi alpha, and Gcommon alpha with a higher affinity to the antibody against Gs alpha. Immunoreaction with the anti-Gs alpha-antibody was markedly higher in pathogenic strains of E. coli. Because of the conserved structure in all GTP-binding proteins which seem to derive from a single primordial protein involved in signal transduction mechanisms, it is reasonable to assume that some anti-Ga-positive proteins in E. coli might be related to G-proteins of higher organisms. A putative candidate for bacterial G-proteins seems to be a 36 kDa protein. Enhancement in G-protein immunostaining in the cytoplasm of macrophages around the internalized bacteria testifies to the involvement of G-proteins in mediation of endocytosis responses of phagocytes.     

Influence of doping and substrate type on the metastable phase growth in pulsed-laser evaporated PbTe films

PbTe films, undoped and doped with 0.02, 0.3 mol% Cr and 0.02 mol% Co, grown by pulsed-laser evaporation (PLE) on different types of substrates have been investigated. The crystal structure of the films was studied by X-ray diffraction. Along with the stable fcc phase, metastable GeS- and CsCl-type phases of PbTe were detected in the films. The growth of the metastable phases as well as the influence of the substrate type and temperature and the doping on the growth have been analyzed in the framework of the classical thermodynamic theory of crystallization. It was found that the influence of doping consists in the change of the surface free energy of the deposited material, while the substrate-type influence depends on its linear thermal expansion coefficient-when it is equal to that of the deposited compound the substrate stimulates the growth of the metastable phases. It is concluded that the growth mechanism is graphoepitaxy.     

Noninvasive real-time multipoint temperature control for ultrasoundphased array treatments

A method for noninvasively estimating spatiotemporal temperature changes in samples using diagnostic ultrasound, and using these as inputs to a multipoint ultrasound phased array temperature controller, is presented in this paper. This method is based on a linear relationship between the apparent tissue echo pattern displacements and temperature, as seen along A-lines acquired with diagnostic ultrasound when the sample is heated by external heating fields. The proportionality constant between echo displacement and temperature is determined by the local change in speed of sound due to temperature and the linear coefficient of thermal expansion of the material. Accurate estimation of the displacements and proportionality constant yields accurate calibrated high-resolution (1 mm spatial, sub-°C) noninvasive sample temperature estimates. These are used as inputs to a multipoint temperature controller, capable of controlling ultrasound phased array treatments in real-time. Phantom and in vitro results show that the noninvasively estimated temperature values can effectively be used to control ultrasound hyperthermia treatments, almost replacing invasive thermocouple measurements. The mathematical background and assumptions of the noninvasive temperature estimator and the controller are presented in this paper, together with experimental results showing estimator and controller performance and limitations. To the best of our knowledge, this paper presents the first demonstration of real-time treatment control based entirely on noninvasive temperature estimates     

A vibrating-tube densimeter for fluids at high pressures and temperatures

A vibrating U-tube apparatus has been developed for determining the densities of pure fluids and fluid mixtures at 10-200 MPa and 323-773 K. Measured parameters areP,T, andr (period of vibration). Fluids are injected into the U-tube at constantP andT. Three or more reference fluids are used to calibrate the response of the instrument. Fluid mixtures are produced by pumping pure fluids into T-junctions on the upstream side of the U-tube using high accuracy syringe pumps. An automated syringe pump is used to maintainP at setpoint ±0.01 MPa.T is controlled to ±0.01 K using a closed-loop, electronic signal amplification/feedback system. For mixtures, a statistically significant number of measurements of r are obtained to account for the effects of small heterogeneities in fluid composition (generally <0.005X;). Typically, density data for 15 fluids can be obtained in a 6- to 8-h period. Considering all of the potential sources of error in the experimentation, conservative estimates of uncertainty are as follows:P, ±0.02 MPa;T, ±0.05 K;p (pure fluids), ±0.0005g.cm^–3; andp (fluid mixtures), ±0.0005-0.0010g-cm^–3.Paper presented at the Twelfth Symposium on Thermophysical Properties, June 19–24, 1994, Boulder, Colorado, U.S.A.     

A histological and histomorphometrical evaluation of screw-type calciumphosphate (Ca-P) coated implants; an in vivo experiment in maxillary cancellous bone of goats

The bone response to different calcium phosphate (Ca-P) coated implants was evaluated in a goat animal model. Two types of plasma spray coatings were applied to a commercially pure titanium (cpTi) tapered, conical screw-design implant (BioComp^®); hydroxyapatite (HA-PS) and a dual coating, consisting of FA and HA (FA/HA-PS). In addition an amorphous RF magnetron sputter coating (Ca-P-a) and uncoated implants were investigated. Forty-eight implants were inserted in the maxilla of 12 adult female goats. After implantation periods of 3 and 6 months, the bone implant interface was evaluated histologically and histomorphometrically. After both implantation periods all plasma spray coated implants were maintained. On the other hand three Ca-P-a and two cpTi implants were lost. Histological examination revealed a better bone response to both plasma spray coated implants. Histomorphometrical evaluation confirmed this finding. At 3 and 6 months significantly higher percentages of bone contact (p<0.001, ANOVA) were measured for both plasma spray coated implants than for the cpTi and Ca-P-a implants, while no significant difference (p<0.05) existed between both implantation periods. Degradation of both plasma spray coatings was observed. Supported by the results, it is concluded that, although Ca-P coatings can improve the performance of dental implants, the presence of a Ca-P coating is not the only important factor for bone healing around implants placed in low density trabecular bone.     

Reprocessing of Microcrystalline Cellulose Spheres with Low Drug Concentrations

Spherical dosage forms have been reported to be an efficient and effective method for delivering drugs into the body and controlling their dissolution rate. Substantial work has been conducted in these laboratories illustrating the advantages of microcrystalline cellulose-based spheres for these purposes. Through various methodologies, but most routinely the extrusion and marumerization technique, it has been determined by Funck, et al. that not more than 50% drug can be incorporated into a sphere formulation without the addition of other binders.

Because of the nature of the extrusion and marumerization manufacturing process, the type of drug being processed and the resultant particle size requirements of the spheres, the percent of spheres falling outside the desired particle size can range from between 3% and 20%. For this reason and the possibility of operator errors, our objective was to determine the parameters which needed monitoring when reprocessing was necessary.     

A terrain parabolic equation model for propagation in thetroposphere

A method to model tropospheric radiowave propagation over land in the presence of range-dependent refractivity is presented. The terrain parabolic equation model (TPEM), is based on the split-step Fourier algorithm to solve the parabolic wave equation, which has been shown to be numerically efficient. Comparisons between TPEM, other terrain models (SEKE, GTD, FDPEM), and experimental data show predominantly excellent agreement. TPEM is also compared to results from an experiment in the Arizona desert in which range-dependent refractive conditions were measured. Although horizontal polarization is used in the implementation of the model, vertical polarization is also discussed     

Effect of internal heating during hot compression on the stress-strain behavior of alloy 304L

The temperature change due to the conversion of mechanical deformation to internal heat and its effect on the as-measured stress-strain behavior of alloy 304L was investigated by means of initially isothermal (compression specimen, dies, and environment at same temperature at initiation of test), constant strain rate, uniaxial compression of laboratory-sized cylindrical specimens. Strain rate was varied in the range 0.01 to 1 s⁻¹, where the thermal state of the test specimen varied from nearly isothermal to nearly adiabatic, respectively. Specimens were deformed in the temperature range of 750 °C to 1150 °C to a strain of 1. The change in specimen temperature with applied strain was calculatedvia finite-element analysis (FEA) from the asmeasured stress-strain data. Selected predictions were confirmed with embedded thermocouples to verify the model employed. Temperature was found to increase monotonically with strain at a strain rate of 1 s^-1, consistent with what is theoretically expected for the adiabatic case. At the 0.1 and 0.01 s^-1 rates, the sample temperature initially increased, peaked, and then decreased as the sample thinned and the contact area between the sample and the cooler dies increased. As-measured stress was corrected for softening associated with deformational heating by interpolation between the various instantaneous stress-temperature behaviors. The resulting isothermal flow data are compared to those predicted by a conventional method that employs an empirical estimate of the heat retention efficiency of the test specimen, assumed dependent on strain rate but independent of strain, to reduce the increase in temperature calculated for the adiabatic case. Differences between the calculated isothermal stress-strain data from the two methods are discussed. Values for the apparent activation energy of deformation and the strain to the peak in the flow curve, which is associated with the onset of dynamic recrystallization, determined from isothermal stress-strain data differed significantly from those obtained from the as-measured compression test data. Formerly Senior Systems Engineer with EG&G Rocky Flats, Inc., is retired.