A new transducer for facet force measurement in the lumbar spine: benchmark and in vitro test results.

A new transducer capable of direct measurement of time-dependent loads in human lumbar facet joints was developed and tested. The transducer was comprised of a force-sensitive resistor (FSR) in series with a pressure-sensitive film. A wide range of experiments revealed the performance attributes and limitations of the FSR. The output signal of the FSR is actually sensitive to both force and area of contact independently. Therefore, a pressure-sensitive film was used to quantify the contact area. At least two transformation equations were calculated for each FSR corresponding to known contact areas. Each equation was a linearization of the log of the FSR output vs the log of the applied ramp loads. Coefficients of determination (CD) were calculated for small (21 mm2) and large (32 mm2) contact areas, and were found to exceed 0.900 for all data. The average of nine cycles was nearly linear for some FSRs (CD of 0.999). FSR output signal and contact area were recorded in cadaveric lumbar facets under ramp load. The appropriate transformation equation was determined by a linear interpolation between benchmark equations based on the contact area measured in vitro. Facet force measurements compared well with those of other researchers. The transducer was found to be quite easy to use.     

A pressure distribution transducer for in-vitro static measurements in synovial joints

The basic features and the performance characteristics of a transducer to measure in-vitro static pressure distribution at the articular interfaces of intact synovial joints are described. Pressure distribution is interpreted from the micro-indentation pattern left on a thin plastic material, the indenter and the plastic material having been subjected to load between the articular surfaces. The effects of the finite thickness and compliance of the transducer and the effects of the time-dependent response properties of the articular cartilage on the accuracy of measurement have been estimated by means of specific experiments and analyses.     

A radioimmunoassay for corticosterone and its application to the measurement of stress in poultry.

A radioimmunoassay for corticosterone was developed using an antibody to corticosterone-21-hemisuccinate:bovine serum albumin. The assay possessed good specificity, sensitivity and reproducibility and required minimal sample preparation. Tests of adrenal function showed that stimulation of the adrenal with exogenous ACTH and with dexamethasone caused an increase and decrease, respectively, in plasma concentrations of corticosterone. Exposure to cold environmental temperatures caused an increase in plasma corticosterone. Handling and the removal of blood samples by venepuncture had no effect upon the concentration of corticosterone. It was concluded that this assay would accurately measure the response to stresses which affect the pituitary-adrenal axis.     

A signal transducer for aerotaxis in Escherichia coli.

The newly discovered aer locus of Escherichia coli encodes a 506-residue protein with an N terminus that resembles the NifL aerosensor and a C terminus that resembles the flagellar signaling domain of methyl-accepting chemoreceptors. Deletion mutants lacking a functional Aer protein failed to congregate around air bubbles or follow oxygen gradients in soft agar plates. Membranes with overexpressed Aer protein also contained high levels of noncovalently associated flavin adenine dinucleotide (FAD). We propose that Aer is a flavoprotein that mediates positive aerotactic responses in E. coli. Aer may use its FAD prosthetic group as a cellular redox sensor to monitor environmental oxygen levels.     

A fluorometric assay for the measurement of endothelial cell density in vitro

Summary A fluorometric assay for determining endothelial cell numbers based on the endogenous enzyme acid phosphatase is described. In preliminary studies, three substrates—p-nitrophenyl phosphate, 4-methylumbelliferyl phosphate, and 2′-[2-benzthiazoyl]-6′-hydroxy-benthiazole phosphate (AttoPhos™)—were compared with respect to their kinetic, optimum assay conditions, sensitivity, and detection limits. Only AttoPhos™ was found to have a high degree of sensitivity, reliability, and reproducibility for measuring both high and low cell numbers in the same plate. In subsequent experiments, assay conditions were validated for measuring endothelial cell density in response to basic fibroblast growth factor and fumagillin. Furthermore, the AttoPhos™ assay revealed a linear correlation between acid phosphatase activity and cell number in many cell types, including BALB/3T3, CHO-K1, A431, MCF7, 2008, SK-OV-3, T47-D, and OVCAR-3. This assay is potentially valuable for use in many in vitro systems in which the quantitation of cell density and proliferation is necessary. The practical advantages of AttoPhos™ assay for measuring endothelial cell numbers include (1) nonradioactivity, (2) simplicity, (3) economy, (4) speed of assessment of proliferation of large number of samples, and (5) amenability to high-throughput drug screening.     

Development of transducer matrices based upon nanostructured conducting polymer for application in biosensors

The nanostructured polyaniline (NSPANI) and its gold nano composite (GNP) with controlled size distribution were developed using structure directing agents (SDA). The nano structure of polyaniline were investigated by UV-Visible spectroscopy, transmission electron microscopy (TEM), dynamic light scattering (DLS), fourrier transform spectroscopy (FTIR), X-ray crystallography and scanning electron microscopy(SEM) etc. These characterization techniques reveal the spherical shape of polyaniline nanoparticles and size in the range of 7-50 nm depending on the type of dopant and nature of SDA. In general, these NSCP colloidal solutions are highly stable. UV-Visible spectra show mainly two peaks at 360-430 nm and at 780-870 nm. The bathochromic shift of the UV-Visible bands as compared to bulk polyaniline, reflect high DC conductivity. TEM and DLS results demonstrate the formation of nanostructure with narrow size distribution. Due to remarkable properties of, it is used as an efficient transduction matrice for the development of highly sensitive, reproducible, stable optical cholesterol and H202 biosensors having wide range of linearity and low Km values.     

A piezoelectric immunobiosensor for atrazine in drinking water.

A piezoelectric crystal immunobiosensor has been developed for the assay of atrazine herbicides in drinking water. Determinations from 0.03-100 micrograms l-1 (parts per billion) of atrazine can be made with a relative SD of about +/- 8%. Atrazine antibodies (polyclonal from sheep) are layered onto the gold electrode of 10 MHz piezoelectric crystals, which are precoated with protein A. The sensor is reversible, being reusable for about eight or nine assays.     

A possible magnetic transducer in birds.

A growing body of evidence indicates that a variety of animals can sense the earth's magnetic field. The possibility that the transducer of the magnetic sense is a microscopic ferromagnet of the sort observed in certain bacteria is examined theoretically. Neither energy and response time considerations nor existing experimental evidence make such a transducer implausible.     

One-legged stance as a representative static body position for calculation of hip contact stress distribution in clinical studies

It was shown in several clinical studies that static one-legged stance may be a relevant body position to describe the loads acting at the hip. However, the stress distribution averaged during movement may better describe hip load than hip contact stress distribution in the static body position. Using data on the resultant hip force during walking taken from the measurements of Bergmann (2001), spatial distribution of contact stress over the articular surface was calculated by the HIPSTRESS method and compared with the stress distribution in one-legged stance. It is shown, that the shape of the contact stress distribution during one-legged stance closely resembled the averaged contact stress distribution during the walking cycle (Pearson's correlation coefficient R2 equals; .986; p < .001). This finding presents a link between the hypothesis that the averaged contact stress distribution during a walking cycle is crucial for cartilage development and the results of clinical studies in which the calculated distribution of contact stress in one-legged stance was successfully used to predict the clinical status of the hip.     

Computer determination of contact stress distribution and size of weight bearing area in the human hip joint

The mathematical models and the corresponding computer program for determination of the hip joint contact force, the contact stress distribution, and the size of the weight bearing area from a standard anteroposterior radiograph are described. The described method can be applied in clinical practice to predict an optimal stress distribution after different operative interventions in the hip joint and to analyze the short and long term outcome of the treatment of various pathological conditions in the hip. A group of dysplastic hips and a group of normal hips were examined, with respect to the peak contact stress normalized by the body weight, and with respect to the functional angle of the weight bearing area. It is shown that both these parameters can be used in the assessment of hip dysplasia.     

Two-dimensional color-mapping of turbulent shear stress distribution downstream of two aortic bioprosthetic valves in vitro.

Since artificial heart valve related complications such as thrombus formation, hemolysis and calcification are considered related to flow disturbances caused by the inserted valve, a thorough hemodynamic characterization of heart valve prostheses is essential. In a pulsatile flow model, fluid velocities were measured one diameter downstream of a Hancock Porcine (HAPO) and a Ionescu-Shiley Pericardial Standard (ISPS) aortic valve. Hot-film anemometry (HFA) was used for velocity measurements at 41 points in the cross-sectional area of the ascending aorta. Three-dimensional visualization of the velocity profiles, at 100 different instants during one mean pump cycle, was performed. Turbulence analysis was performed as a function of time by calculating the axial turbulence energy within 50 ms overlapping time windows during the systole. The turbulent shear stresses were estimated by using the correlation equation between Reynolds normal stress and turbulent (Reynolds) shear stress. The turbulent shear stress distribution was visualized by two-dimensional color-mapping at different instants during one mean pump cycle. Based on the velocity profiles and the turbulent shear stress distribution, a relative blood damage index (RBDI) was calculated. It has the feature of combining the magnitude and exposure time of the estimated shear stresses in one index, covering the entire cross-sectional area. The HAPO valve showed a skewed jet-type velocity profile with the highest velocities towards the left posterior aortic wall. The ISPS valve revealed a more parabolic-shaped velocity profile during systole. The turbulent shear stresses were highest in areas of high or rapidly changing velocity gradients. For the HAPO valve the maximum estimated turbulent shear stress was 194 N m-2 and for the ISPS valve 154 Nm-2. The RBDI was the same for the two valves. The turbulent shear stresses had magnitudes and exposure times that might cause endothelial damage and sublethal or lethal damage to blood corpuscules. The RBDI makes comparison between different heart valves easier and may prove important when making correlation with clinical observations.     

A miniature yeast telomerase RNA functions in vivo and reconstitutes activity in vitro

The ribonucleoprotein enzyme telomerase synthesizes DNA at the ends of chromosomes. Although the telomerase catalytic protein subunit (TERT) is well conserved, the RNA component is rapidly evolving in both size and sequence. Here, we reduce the 1,157-nucleotide (nt) Saccharomyces cerevisiae TLC1 RNA to a size smaller than the 451-nt human RNA while retaining function in vivo. We conclude that long protein-binding arms are not essential for the RNA to serve its scaffolding function. Although viable, cells expressing Mini-T have shortened telomeres and reduced fitness as compared to wild-type cells, suggesting why the larger RNA has evolved. Previous attempts to reconstitute telomerase activity in vitro using TLC1 and yeast TERT (Est2p) have been unsuccessful. We find that substitution of Mini-T for wild-type TLC1 in a reconstituted system yields robust activity, allowing the contributions of individual yeast telomerase components to be directly assessed.