Experimental infection of pigs with a thymidine kinase negative strain of pseudorabies virus

Sixteen 20 day old pigs, devoid of neutralizing antibody to pseudorabies virus (PRV), were divided into two groups of eight, an the animals of each group were housed in a separate unit. In each group 6 pigs were inoculated intranasally with the thymidine kinase (TK-) mutant (Group 1) or the field strain of PRV (Group 2), each pig receiving an inoculum of 4 ml. The remaining 2 pigs in each group served as uninoculated controls. The only clinical sign observed in the pigs of Group 1 was a transient febrile reaction, in the case of six pigs inoculated with the TK- mutant of PRV, whereas no signs of disease were seen in the uninoculated controls. The virus was isolated from the 6 infected pigs of the group only on post infection day (PID) 2, whereas it was never isolated from the controls. By contrast, the pigs of Group 2, had a severe clinical response and one, among those that were inoculated with the field strain of the PRV, died on PID 9. Virus was consistently isolated from all pigs of Group 2, inoculated and control. On PID 30 all pigs, i.e. the 8 of Group 1 and 7 of the Group 2 which survived to the infection, were subjected to dexamethasone (DMS) treatment. After DMS treatment virus was never isolated from the nasal swabbings obtained from the pigs of Group 1, whereas it was consistently isolated from pigs of Group 2. After 30 d from the start of DMS treatment the pigs were killed and several tissues were collected from each pig for virus detection, by isolation in tissue culture and by PCR analysis. At necropsy no lesions were found in pigs of Group 1, whereas acute pneumonia and gliosis in the trigeminal ganglia were observed in pigs of Group 2. Virus was never isolated from any of the tissues taken from pigs of both, Group 1 and Group 2, nevertheless sequences of PRV were detected by PCR analysis in the trigeminal ganglia of the pigs of both Groups.     

In vitro evaluation of electrostatic endothelial cell transplantation onto 4 mm interior diameter expanded polytetrafluoroethylene grafts

PURPOSE: To perform an in vitro evaluation of electrostatic endothelial cell transplantation of human umbilical vein endothelial cells (HUVEC) onto segments of 4 mm internal diameter expanded polytetrafluoroethylene (ePTFE) vascular prostheses. METHODS: This evaluation consisted of exposing vascular graft segments that had been subjected to either electrostatic or gravitation transplantation with HUVEC to a physiologic shear stress (15 dynes/cm2) under steady flow conditions within a flow loop system. Biochemical assays were performed on freshly transplanted grafts by means of radioimmunoassay for prostacyclin and thromboxane A2. RESULTS: There was a 30% loss of HUVEC after 30 minutes of shear stress exposure from the grafts subjected to gravitational transplantation with no additional significant (alpha = 0.05) loss after 120 minutes. Grafts subjected to electrostatic transplantation had no significant (alpha = 0.05) loss of HUVEC during exposure to physiologic shear stress. Furthermore, after 120 minutes of shear-stress exposure, the grafts subjected to electrostatic transplantation (78,420 +/- 6274 HUVEC/cm2) retained 2.3 times more HUVEC than the counterparts subjected to gravitational transplantation (34,427 +/- 4637 HUVEC/cm2). The biochemical assay results indicated no significant (alpha = 0.05) production of prostacyclin or thromboxane A2 regardless of the method of cell transplantation. CONCLUSIONS: (1) The electrostatic transplantation technique was superior to the gravitational transplantation technique in terms of cellular retention when the ePTFE grafts were exposed to physiologic shear stress. (2) Production of prostacyclin and thromboxane A2 did not differ between transplanted HUVEC subjected to gravitational or electrostatic procedures.     

An analytical technique for the determination of spectral components of multilevel carrier-based PWM methods

This paper presents an analytical approach for determining the spectral components of multilevel pulsewidth modulation (PWM) strategies, which avoids the problems of spectral leakage, roundoff, and subtle programming errors associated with the more usual approach of using discrete Fourier transforms of simulated switched waveforms. The approach uses a double Fourier integral formulation to analyze the inverter switched output, establishing the inner and outer integral limits of this formulation under conditions of natural sampling, regular sampling, and overmodulation. Closed-form and numerical integration solutions are presented for phase disposition PWM of a diode-clamped inverter to illustrate the technique, together with a comparison of theoretical and experimental spectral results for the naturally and asymmetric regularly sampled cases to confirm the validity of the approach.     

A 1.1-V 270-/spl mu/A mixed-signal hearing aid chip

This paper presents a single-chip mixed-signal IC for a hearing aid system. The IC consumes 270 /spl mu/A of supply current at a 1.1-V battery voltage. The presented circuit and architectural design techniques reduce the total IC power to 297 /spl mu/W, a level where up to 70 days of lifetime is achieved at 10 h/day for a small zinc-air battery. The measured input referred noise for the entire channel is 2.8 /spl mu/Vrms and the average THD in the nominal operating region is 0.02%. The jitter for the on-board ring oscillator is 147 ps rms. The chip area is 12 mm/sup 2/ in a 0.6-/spl mu/m 3.3-V mixed-signal CMOS process.     

The influence of energy intake on the pathophysiology of Trypanosoma congolense infection in Scottish blackface sheep

The intensity of parasitaemia, degree of anaemia, live body weight gains and blood biochemical changes were measured in two groups of Scottish Blackface sheep infected experimentally with Trypanosoma congolense and allowed either a high (9.9 MJ metabolisable energy (ME) day-1) or a low (6.1 MJ ME day-1) energy intake. It was observed that infected animals on the low energy intake had a longer mean prepatent period, but following patency they developed more severe anaemia and greater growth retardation than those on the high energy intake. Both infected groups exhibited significant reductions in serum total lipids, phospholipids, plasma cholesterol and albumin. However, these changes were more severe in the animals on the low energy intake than in those on the high energy intake. It was concluded that adequate energy nutrition enhances the ability of infected animals to withstand the adverse effects of infection, by promoting body weight gains and moderating the severity of the pathophysiological changes associated with ovine trypanosomosis.     

Purification by reflux electrophoresis of whey proteins and of a recombinant protein expressed in Dictyostelium discoideum

Protein purification that combines the use of molecular mass exclusion membranes with electrophoresis is particularly powerful as it uses properties inherent to both techniques. The use of membranes allows efficient processing and is easily scaled up, while electrophoresis permits high resolution separation under mild conditions. The Gradiflow apparatus combines these two technologies as it uses polyacrylamide membranes to influence electrokinetic separations. The reflux electrophoresis process consists of a series of cycles incorporating a forward phase and a reverse phase. The forward phase involves collection of a target protein that passes through a separation membrane before trailing proteins in the same solution. The forward phase is repeated following clearance of the membrane in the reverse phase by reversing the current. We have devised a strategy to establish optimal reflux separation parameters, where membranes are chosen for a particular operating range and protein transfer is monitored at different pH values. In addition, forward and reverse phase times are determined during this process. Two examples of the reflux method are described. In the first case, we described the purification strategy for proteins from a complex mixture which contains proteins of higher electrophoretic mobility than the target protein. This is a two-step procedure, where first proteins of higher mobility than the target protein are removed from the solution by a series of reflux cycles, so that the target protein remains as the leading fraction. In the second step the target protein is collected, as it has become the leading fraction of the remaining proteins. In the second example we report the development of a reflux strategy which allowed a rapid one-step preparative purification of a recombinant protein, expressed in Dictyostelium discoideum. These strategies demonstrate that the Gradiflow is amenable to a wide range of applications, as the protein of interest is not necessarily required to be the leading fraction in solution.     

Realization of novel low-loss monolithically integrated passive waveguide mode converters

The novel design and fabrication of monolithically integrated passive waveguide mode converters (WMCs), realized through the utilization of the reactive ion-etch lag (RIE Lag) phenomenon, is reported. The low-loss GaAs-AlGaAs WMCs have been characterized over a wavelength range of 900-940 nm, resulting in TE-TM (TM-TE) mode conversion with efficiencies of greater than 96% and low-loss devices with conversion lengths as short as 150 /spl mu/m. The properties and characteristics of the fully integrated WMCs, fabricated with a single masking and etch process, are in agreement with theoretical predictions.