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PubMed1992

Utility of immobilon-bound phosphoproteins as substrates for protein phosphatases from neutrophils.

Immobilon-bound phosphoproteins labeled with 32P were utilized as substrates to study the enzymes in neutrophils that are active against the major products of protein kinase C. The labeled proteins were separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and transferred electrophoretically to immobilon-P membranes. Both particulate and soluble phosphatases were found to be active against the blotted phosphoproteins. Reactions were followed by autoradiography as the loss of 32P from individual protein bands. The tumor promoter okadaic acid and the hepatoxin microcystin-LR inhibited these reactions in a manner consistent with the enzymes being type 1 and/or 2A protein phosphatases.

Animals↗
PubMed1983

Pathologic findings in rats following inhalation of combustion products of polytetrafluoroethylene (PTFE).

Adult male Fischer 344 rats received single 30-min exposures to the aerosolized products of polytetrafluoroethylene (PTFE) heated to 595 degrees C. The concentrations of thermal degradation products of PTFE were at the LC50 dose of 0.045 mg/l for most rats, but some rats received doses ranging from 0.005 to 5.025 mg/l. Serial measurements of cardiopulmonary function were obtained and will be published subsequently. Necropsies were performed at 0, 2, 12, 24 and 36 h post-exposure, and a few rats were killed between 2 and 17 days. Signs of respiratory impairment were followed by death in some rats. Pathologic findings included focal hemorrhages, edema and fibrin deposition in the lungs. With time focal interstitial thickenings developed due to hypertrophy and hyperplasia of alveolar cells, and macrophages accumulated in alveoli. Thrombosis of pulmonary capillaries was common. Disseminated intravascular coagulation (DIC) occurred in 53% of test rats; its incidence and severity were positively related to the degree of pulmonary damage. Renal infarcts were common due to DIC. No lesions were seen in kidneys or other tissue (except lung and thymus) unless they were affected with DIC. Thymic lymphocytes underwent necrosis in many test and some vehical (warm air) control rats, possibly due to stress. The finding of DIC in PTFE combustion product exposure has not been reported to our knowledge. The toxicity of the thermal degradation products of PTFE requires further study, especially relative to induction of DIC.

Animals↗
PubMed1975

Determination of fluorocarbon 11 and fluorocarbon 12 in post-mortem tissues: a case report.

This report describes the death of a teenager due to inhalation of fluorocarbon aerosol propellants and presents a method for the determination of trichlorofluoromethane (fluorocarbon 11) and dichlorodifluoromethane (fluorocarbon 12) in post-mortem samples. The post-mortem blood and tissue levels of these fluorocarbons are also presented. The distribution of fluorocarbon 11 and fluorocarbon 12 is similar to that observed in chloroform deaths.

Adolescent↗
PubMed1992

The influence of spatial polarization distribution on spot poled PVDF membrane hydrophone performance.

The influence of spatial polarization distribution on spot poled PVDF membrane hydrophone performance was calculated and then experimentally verified using a one-dimensional model based on acoustic wave propagation through a layered medium. It is shown that the sensitivity of the hydrophone is markedly dependent on the spatial polarization distribution. It is also shown that there can be a significant difference between the voltage sensitivities measured in the same hydrophone probe depending on which electrode is actually facing the acoustic source. The measurements carried out in the frequency range 1-20 MHz indicate that this difference, while negligible below 2 MHz, may exceed 1.6 dB at higher frequencies. The model developed can also be used to determine the "effective" piezoelectric constant d33 of the PVDF material as a continuous function of frequency. Moreover, the model predicts the existence of a negative slope in the frequency response of the spot poled membrane hydrophone. The experimental confirmation of this prediction underscores the importance of using swept frequency methods during calibration measurements.

Electricity↗
PubMed1995

Influence of water temperature on pressure pulses generated by an electromagnetic type lithotripter.

The need to survey and compare in vitro investigations on the effect of extracorporeally generated pressure pulses demands precise description of experimental conditions. We determined the effect of water temperature on the pressure characteristics in and around the focus area of an electromagnetic type lithotripter. The peak pressure amplitude at the geometrical focus of the lens was reduced to 60% of the original peak pressure if water at room temperature was used instead of water with human body temperature. By decreasing the temperature of the lithotripter's internal water, we observed shifting of the focus point at a rate of 1.4 mm per degree. This dramatic reduction of the peak pressure and the shifting of the focus point resulting from changes in the acoustic and thermal parameters of the propagation medium and of the focusing device could lead to differences of up to 80% of peak pressure amplitude.

Acoustics↗
PubMed1995

A 40-100 MHz B-scan ultrasound backscatter microscope for skin imaging.

There is a growing interest in high resolution, subsurface imaging of cutaneous tissues using higher frequency ultrasound, and several commercial systems have been developed recently which operate at 20 MHz. Some of the possible applications of higher frequency skin imaging include tumour staging, boundary definition, and studies of the response of tumours to therapy, investigations of inflammatory skin conditions such as psoriasis and eczema, and basic studies of skin aging, sun damage and the effects of irritants. Investigation of these areas is quite new, and the role of ultrasound skin imaging is continuing to evolve. Lateral resolution in the 20 MHz imaging systems ranges from 200 to 300 microns, which limits imaging applications to cutaneous structures which are relatively large in size. In this paper, a real-time ultrasound backscatter microscope (UBM) for skin imaging is described which operates in the 40-100 MHz range, providing axial resolution between 17 and 30 microns and lateral resolution between 33 and 94 microns. This improvement in resolution over current skin ultrasound systems should prove useful in determining the margins of small skin lesions, and in obtaining more precise, in vivo skin thickness measurements to characterize nonmalignant skin disease. Example images of normal skin, seborrhoeic keratosis and malignant melanoma illustrate the imaging potential of this system.

Analog-Digital Conversion↗
PubMed1995

Damage to red blood cells induced by acoustic cavitation.

This experimental study has revealed damage to red blood cells that is quantitatively related to the acoustic pressure during irradiation with 0.75-MHz continuous-wave ultrasound, using a range of intensities comparable to those employed by ultrasonic physiotherapy equipment (0.25 to 7 W cm-2 spatial average temporal average). Damage to the red blood cells was investigated by measuring the extent of haemolysis using a UV/VIS spectrophotometer. There was a clear correlation between the amount of haemoglobin released and the intensity of harmonic emissions recorded from the sample during irradiation. The observed degree of haemolysis could not be produced by temperature rises in the absence of the sound field. This suggests that the damage was a direct result of acoustic cavitation.

Acoustics↗
PubMed1995

Importance of the implosion of ESWL-induced cavitation bubbles.

The damage induced by an extracorporeal shock wave lithotripter is observed with a fiber optic stress sensing technique. When a stone is placed in the focus, besides the expected stress induced by the incoming shock wave emitted by the ESWL apparatus, a second delayed stress is observed some hundreds of microseconds later. The second stress is induced by a shock wave generated at the collapse of a cavitation bubble. Partial reflection of the incoming shock wave at the stone boundary is at the origin of the large cavitation bubble formation. Sensing fiber fracture results always from the second shock wave due to the collapse of the cavitation bubble. Thus the largest stress is generated at the collapse. When no target is placed in the focus of the lithotripter, no large cavitation bubble is formed and no delayed shock wave is observed. Our results demonstrate unambiguously the decisive role of cavitation in ESWL procedures.

Acoustics↗
PubMed1988

Measurement of extracellular basal levels of serotonin in vivo using nafion-coated carbon fibre electrodes combined with differential pulse voltammetry.

Carbon fibre electrodes combined with differential pulse voltammetry have been used for a number of years to monitor changes in the extracellular concentrations of ascorbic acid, dihydroxyphenylacetic acid, and 5-hydroxyindoleacetic acid. However, the primary objective of in vivo electrochemists has been to monitor changes in the extracellular concentrations of the neurotransmitter amines; dopamine and serotonin rather than their metabolites. In this paper we describe a new chemically- and electrically-pretreated Nafion-coated carbon fibre electrode which can be used to monitor basal levels of serotonin in the extracellular fluid in the frontal cortex and the dorsal raphe nucleus of rat. These electrodes combined with differential pulse voltammetry detect dopamine (Peak A at -70 mV) and serotonin (Peak B at +240 V) oxidation peaks in vitro but not the oxidation of ascorbic acid, dihydroxyphenylacetic acid, 5-hydroxyindoleacetic acid or uric acid, at concentrations up to 10 microM. These electrodes were able to detect serotonin concentration as large as 1 nM in vitro. When used in vivo the oxidation peaks obtained in the frontal cortex and dorsal raphe indicate the basal concentrations of serotonin to be 5 nM and 10 nM respectively. Pharmacological interventions in rats implanted with normal carbon fibre electrodes or with Nafion carbon fibre electrodes further demonstrate that the new Nafion electrodes measure serotonin in vivo. The Nafion-coated electrodes therefore may be a useful tool for the study of serotoninergic systems in vivo with the added advantage that they cause minimal damage due to their small tip size (30 micron).

Animals↗
PubMed1993

A hydromechanical model of the cochlea with nonlinear feedback using PVF2 bending transducers.

A single-channel hydromechanical model of the human cochlea with nonlinear feedback on the membrane is introduced. The piezoelectric transducers for displacement measurement and force exertion on the membrane are constructed of PVF2 in the bimorph bending type. The passive response shows high-frequency slopes of about 60 dB/oct and a shortest wavelength of about 17 mm corresponding to 0.9 Bark in the human cochlea. High-frequency slopes and Q10 dB with one feedback loop are within the range of animal data, where as dynamic compression does not exceed 20 dB. Input impedance and phase response are in good agreement with nature, which is ascribed to longitudinal stiffness of the membrane.

Acoustic Stimulation↗
PubMed1994

Application of a Nafion-modified carbon paste electrode for the absorptive stripping voltammetric determination of fenoterol in pharmaceutical preparations and biological fluids.

The preconcentration of fenoterol on a Nafion-modified carbon paste electrode and its subsequent determination using differential pulse voltammetry is described. The effect of pH and percentage Nafion concentration on the accumulation behaviour of fenoterol was studied, and accumulation curves, calibration graphs and reproducibility studies at two different Nafion concentrations have been carried out in the range 2.5 x 10(-8)-5.0 x 10(-7) M fenoterol. A limit of detection in aqueous solutions, calculated using a signal-to-noise ratio (S/N) of 3, was 9.0 x 10(-9) M. Application of the electrode to pharmaceutical preparations, without sample pretreatment, resulted in acceptable deviation from the stated concentration (RSD = +/- 3.81%, n = 4). For more complex matrices, a suitable extraction procedure was developed, resulting in recoveries of > 90% (urine) and > 75% (serum).

Adsorption↗
PubMed1995

Electrochemical behaviour of clenbuterol at Nafion-modified carbon-paste electrodes.

A detailed study of the electrochemistry of clenbuterol at bare carbon-paste electrodes (CPEs) has been carried out. Results showed that clenbuterol undergoes an ECE process. This compound is irreversibly oxidised at high potentials, resulting in the formation of a product which demonstrates quasi-reversible electrochemical behaviour at less positive potentials. The amount of this chemical product formed is very pH-dependent. Investigations into the electrochemical behaviour of clenbuterol at Nafion-modified CPEs were also made. The use of a thin Nafion film cast over the CPE resulted in a large increase in peak current over bare electrodes. Linear accumulation occurred with time, the linear range increasing with decreasing concentration. This allowed the detection of low concentrations of clenbuterol. Diffusion proved to be the rate-controlling process of clenbuterol through the Nafion membrane.

Carbon↗
PubMed1996

Proton fluorine wideline separation experiments on poly (vinylidene difluoride).

Two-dimensional heteronuclear proton-fluorine correlation solid-state spectroscopy is demonstrated on a sample of poly (vinylidene difluoride) (PVDF). This allows the correlation of structural information contained in the fluorine chemical shift in one dimension with information on mobility from proton wideline spectra in the second dimension. The effect of the heteronuclear fluorine-proton dipolar coupling on the proton linewidth is investigated separately. The incorporation of magnetization filters into the preparation period enables one to measure the wideline separation (WISE) spectra for the amorphous or the crystalline parts of the sample separately.

Fluorine↗
PubMed1993

Chemical protein sequencing and amino acid analysis.

Chemical protein sequencing remains the cornerstone of protein structural analysis. Recent developments allow an increased rate of sampling and a reduction in background noise. Mass spectrometric detection of chemical sequencing products will, in all likelihood, initiate an era of sequencing of sub-picomole quantities of protein. Significant progress toward routine, automated carboxy-terminal sequencing has been reported recently.

Amino Acid Sequence↗
PubMed1995

A protein complex from Choristoneura fumiferana gut-juice involved in the precipitation of delta-endotoxin from Bacillus thuringiensis subsp. sotto.

A 75 kDa protein from spruce budworm (Choristoneura fumiferana) gut-juice has been isolated and shown to cause a specific precipitation of the delta-endotoxin from Bacillus thuringiensis subsp. sotto. This 75 kDa protein, separated by either column chromatography or SDS-PAGE, caused precipitation of the sotto toxin in both agarose diffusion gels and the PAGE gels. The precipitation event leads to limited proteolysis of the toxin and loss of larval toxicity. SDS-PAGE analysis of the precipitated toxin indicates that proteolysis of the toxin is not a prerequisite for precipitation. The protein responsible for precipitation, exhibits elastase-like activity and appears to be a complex which partially dissociates during boiling in SDS-PAGE sample buffer. Gut-juice from gypsy moth, forest tent caterpillar and white mark tussock moth also precipitated delta-endotoxin, but silkworm gut-juice gave a much weaker response. These results provide further evidence that, in the larval gut, differential processing of delta-endotoxin may play a role in the expression of activity towards various insect larvae.

Amino Acid Sequence↗
PubMed1991

Deproteination of nucleic acids by filtration through a hydrophobic membrane.

To remove or inactivate an enzyme from DNA in multistep procedures in molecular biology, it is often necessary to phenol extract the solution, followed by chloroform extraction and ethanol precipitation. In addition to being time-consuming and hazardous, there can be significant loss of DNA with this procedure, especially when small volumes or amounts of DNA are being used. We have found that filtering analytical reaction mixtures through a hydrophobic membrane specifically to remove protein is a rapid alternative to phenol extraction. Within broad limits commonly encountered in molecular biology, filtration through a polyvinylidene difluoride membrane quantitatively removes a variety of enzymes without significant loss of double-stranded nucleic acid.

Alkaline Phosphatase↗