RESUMEN
New ideas for diagnostics in clinical parasitology are needed to overcome some of the difficulties experienced in the widespread adoption of detection methods for gastrointestinal parasites in livestock. Here we provide an initial evaluation of the performance of a newly developed automated device (Telenostic) to identify and quantify parasitic elements in fecal samples. This study compared the Telenostic device with the McMaster and Mini-FLOTAC for counting of strongyle eggs in a fecal sample. Three bovine fecal samples were examined, in triplicate, on each of the three fecal egg-counting devices. In addition, both manual (laboratory technician) and automated analysis (image analysis algorithm) were performed on the Telenostic device to calculate fecal egg counts (FEC). Overall, there were consistent egg counts reported across the three devices and calculation methods. The Telenostic device compared very favourably to the Mini-FLOTAC and McMaster. Only in sample C, a significant difference (P < 0.05) was observed between the egg counts obtained by Mini-FLOTAC and by the other methods. From this limited dataset it can be concluded that the Telenostic-automated test is comparable to currently used benchmark FEC methods, while improving the workflow, test turn-around time and not requiring trained laboratory personnel to operate or interpret the results.
Asunto(s)
Pruebas Diagnósticas de Rutina/veterinaria , Ganado/parasitología , Animales , Bovinos , Pruebas Diagnósticas de Rutina/métodos , Heces/parasitología , Helmintiasis Animal , Enfermedades de los Caballos/parasitología , Caballos/parasitología , Parasitosis Intestinales , Recuento de Huevos de Parásitos/veterinaria , Ovinos/parasitología , Enfermedades de las Ovejas/parasitologíaRESUMEN
The imaging of shear-mediated dynamic platelet behavior interacting with surface-immobilized von Willebrand factor (vWF) has tremendous potential in characterizing changes in platelet function for clinical diagnostics purposes. However, the imaging output, a series of images representing platelets adhering and rolling on the surface, poses unique, non-trivial challenges for software algorithms that reconstruct the positional trajectories of platelets. We report on an algorithm that tracks platelets using the output of such flow run experiments, taking into account common artifacts encountered by previously-published methods, and we derive seven key metrics of platelet dynamics that can be used to characterize platelet function. Extensive testing of our method using simulated platelet flow run data was carried out to validate our tracking method and derived metrics in capturing key platelet-vWF interaction-dynamics properties. Our results show that while the number of platelets present on the imaged area is the leading cause of errors, flow run data from two experiments using whole blood samples showed that our method and metrics can detect platelet property changes/differences that are concordant with the expected biological outcome, such as inhibiting key platelet receptors such as P2Y1, glycoprotein (GP)Ib and GPIIb/IIIa. These findings support the use of our methodologies to characterize platelet function among a wide range of healthy and disease cohorts.
Asunto(s)
Plaquetas , Rastreo Celular/métodos , Procesamiento de Imagen Asistido por Computador/métodos , Pruebas de Función Plaquetaria/métodos , Factor de von Willebrand , Adenosina Difosfato/análogos & derivados , Adenosina Difosfato/farmacología , Algoritmos , Plaquetas/citología , Plaquetas/efectos de los fármacos , Plaquetas/fisiología , Humanos , Microscopía Fluorescente , Microscopía por Video/métodos , Inhibidores de Agregación Plaquetaria/farmacología , Factor de von Willebrand/metabolismo , Factor de von Willebrand/fisiologíaRESUMEN
Age is a risk factor for cardiovascular disease (CVD), however the effect of age on platelet function remains unclear. Ideally, platelet function should be assayed under flow and shear conditions that occur in vivo. Our study aimed to characterise the effect of age on platelet translocation behaviour using a novel flow-based assay that measures platelet function in less than 200 µl of blood under conditions of arterial shear. Blood from males (n = 53) and females (n = 56), ranging in age from 19-82 and 21-70 respectively were perfused through custom-made parallel plate flow chambers coated with immobilised human von Willebrand Factor (VWF) under arterial shear (1,500 s(-1)). Platelet translocation behaviour on VWF was recorded by digital-image microscopy and analysed. The study showed that aging resulted in a significant decrease in the number of platelet tracks, translocating platelets and unstable platelet interactions with VWF. These age related changes in platelet function were more profound in women than in men indicating that age and gender significantly impacts on platelet interactions with VWF.