Shear Force Generating Device for Analysis of Biopsied Mammalian Samples

Abstract

A shear force generating device includes a shear-force generator structured for exerting a shear force at to each volume element of a sample of body fluid or tissue and a native conformation prion protein arranged in a container. A controller controls the shear-force generator to exert a shear-force of at maximum 10% of one shear-force intensity to each volume element of the sample arranged in the container. A light source is directed into the container. An optical detector is arranged to detect light emanating from the container. The detector is coupled to a computer including code to analyze light detected by the optical detector as a measure for the content of aggregated prion protein by comparing the light detected to a Reference Information Database.

1 . Device for analysis of a biopsied mammalian sample of body fluid or tissue and a native conformation prion protein to determine the presence of disease-related aggregated conformation prion protein, the device comprising
 a shear-force generator structured for exerting a shear force to each volume element of the sample arranged in a container,   a controller to control the shear-force generator to exert the shear-force of at maximum 10% of one shear-force intensity to each volume element of the sample arranged in the container,   a light source, the beam path of which is directed into the container,   an optical detector arranged to detect light emanating from the container, wherein the optical detector is a luminescence detector, a scatter light detector, or an absorbance detector,   a computer including code to analyze light detected by the optical detector as a measure for the content of aggregated prion protein by comparing the light detected to a Reference Information Database containing pre-determined data relating detected light to content of aggregated conformation prion protein in relation to specific shear-force intensities and in relation to a diagnosis for a neurodegenerative disease pertaining to a reference sample used for generating the pre-determined data.   
     
     
         2 . The device according to  claim 1 , wherein the Reference Information Database comprises data on amounts of aggregated conformation prion protein, which data are pre-determined for one specific shear-force intensity for a reference sample from a specific source of known diagnosis. 
     
     
         3 . The device according to  claim 1 , wherein the code compares the data in the Reference Information Database to data representing amounts of aggregated conformation prion protein generated from an admixture of sample and native conformation prion protein, wherein the comparison of amounts of aggregated conformation prion protein is made for the same shear-force intensity. 
     
     
         4 . The device according to  claim 1 , wherein the Reference Information Database comprises data for a specific neurodegenerative disease and/or a subtype thereof in association with pre-determined data on amounts of aggregated conformation prion protein, each generated at one of at least two, e.g. 3, 4, 12, 24, or 36 different shear-force intensities for each reference sample. 
     
     
         5 . The device according to  claim 1 , comprising at least two containers, each arranged in a recess formed of a housing containing the optical detector and the light source, a thermostat and a lid section comprising a bearing for an axle carrying the rotor, wherein the housings are connected to one another and the lid sections are connected to one another. 
     
     
         6 . The device according to  claim 1 , wherein the shear-force generator comprises a rotor arranged at a spacing from a stator, wherein the stator has extensions forming a funnel narrowing to an inlet which is coaxial to the rotor. 
     
     
         7 . The device according to  claim 1 , wherein the shear-force generator has a stator formed of a portion of the wall of the container which for a portion of the circumference of the rotor is in parallel to the rotor at a constant spacing. 
     
     
         8 . The device according to  claim 1 , employed for analysis for the presence of disease-related aggregated conformation prion protein in the biopsied mammalian sample by the steps of
 a) adding to the sample at least one native conformation prion protein,   b) via the shear-force generator and controller, subjecting the mixture comprising the sample and at least one native conformation prion protein obtained in step a) to at least one shear-force intensity that is controlled to have a uniform intensity having an intensity range of maximally 20% of one shear-force value for a pre-determined number of cycles of a pre-determined time of shear-force acting and a pre-determined resting phase,   c) with the code, following step b), determining the content of aggregated conformation prion protein for each of the shear-force intensities, characterized by   d) comparing the content of aggregated conformation prion protein determined in step c) to pre-determined data on the content of aggregated conformation prion protein, which content was determined for native conformation prion protein subjected to the same shear-force intensity as in step b), wherein the pre-determined data on the content of aggregated conformation prion protein was determined in native conformation prion protein in admixture with a reference sample and these data are provided in Reference Information Database which in association with these data contains the neurodegenerative disease diagnosis for the patient from which the reference sample originates.   
     
     
         9 . The device according to  claim 8 , wherein comprising, prior to step b) the mixture is divided into aliquots and in step b) at least two aliquots are subjected to a different shear-force intensity each and in step c) the content of aggregated conformation prion protein is determined for each aliquot and in step d) the content of aggregated conformation prion protein determined in step c) for each aliquot is compared to data on a pre-determined content of aggregated conformation prion protein. 
     
     
         10 . The device according to  claim 8 , wherein comprising in step b) the mixture is subjected to a succession of at least two different shear-force intensities and the content of aggregated conformation prion protein is determined during or following subjecting the mixture to each one of the shear-force intensities. 
     
     
         11 . The device according to  claim 8 , wherein comprising adding to the mixture at least one luminescent dye that is specific for the aggregated conformation prion protein prior to the step of subjecting the mixture to at least two different shear-force intensities and measuring the luminescence of the dye. 
     
     
         12 . The device according to  claim 8 , wherein the device is configured for irradiating the mixture with light having a wavelength for exciting luminescence in the dye and measuring the luminescence of the dye during shear-force acting of step b) or during a resting phase of step b), without moving the volume occupied by the mixture relative a the shear-force generator generating the shear-force in step b). 
     
     
         13 . The device according to  claim 8 , configured and controlled for irradiating the mixture with light having a wavelength that is scattered by the aggregated conformation prion protein and measuring scattered light exiting the admixture during step b), or during a pause of step b), with or without moving the volume occupied by the mixture relative a the shear-force generator generating the shear force in step b). 
     
     
         14 . The device according to  claim 8 , wherein in step b) the rate of formation of aggregated conformation prion protein is determined from the content of aggregated state prion protein determined at the at least one shear-force intensity and the pre-determined data contain the rate of formation at the same shear-force intensity. 
     
     
         15 . The device according to  claim 8 , wherein the content of aggregated conformation prion protein is determined as the time-resolved content and that the rate of formation of aggregated conformation prion protein is determined by non-linear regression analysis of an approximation on the determined time-resolved content of aggregated conformation prion protein for each of the shear-force intensities. 
     
     
         16 . The device according to  claim 8 , wherein configured and controlled for adding at least one aggregated conformation prion protein to at least one aliquot of the mixture comprising the sample and at least one native conformation prion protein, wherein the at least one aggregated conformation prion protein is produced by subjecting a native conformation prion protein to a uniform shear-force controlled to an intensity range of maximally 1% of one shear-force intensity. 
     
     
         17 . The device according to  claim 8 , wherein configured and controlled for treating in parallel at least one native conformation prion protein without addition of a sample of body fluid. 
     
     
         18 . The device according to  claim 1 , wherein the Reference Information Database relates the medical diagnosis for a specific neurodegenerative disease and/or a subtype thereof in association with pre-determined data on amounts of aggregated conformation prion protein, which data were generated separately for each of at least two different shear-force intensities for each native conformation prion protein added to the sample. 
     
     
         19 . The device according to  claim 1 , wherein the Reference Information Database includes data of the rate of formation of aggregated conformation prion protein for each of at least two different shear-force intensities, the rate of dissociation from aggregated conformation prion protein and/or the original amount of aggregated conformation prion protein in the sample. 
     
     
         20 . The device according to  claim 1 , wherein the Reference Information Database contains data on pre-determined amounts which are specific for a single shear-force intensity and specific for a seed from a specific source. 
     
     
         21 . The device according to  claim 1 , wherein the Reference Information Database contains the medical diagnosis for a specific neurodegenerative disease and/or a subtype thereof in association with the pre-determined data on amounts of aggregated conformation prion protein, each amount generated at one of at least two different shear-force intensities for each native conformation prion protein added and the diagnosis associated with the sample, and additionally contains the specific source or sample to which the diagnosis pertains. 
     
     
         22 . The device according to  claim 1 , wherein the Reference Information Database contains information on the age of disease onset, sex of the mammal, duration of the disease, progression and/or severity of the disease.