Cell responses to varying electric fields can reveal insights on cell biology with important implications for pharmaceutical and basic research. In this work, we exploit spectral information content ...in Opto-Electronic Tweezers (OET) systems through machine learning for label-free characterization of cell dielectric properties aimed at cell classification and drug response evaluation. A customized Polymethyl-methacrylate (PMMA) chip with ITO substrates and an a-Si layer was designed for OET-based manipulation of cells and integrated with an inverted microscope. We obtained OET cell signatures as spectra responses of kinematic and dynamic descriptors, which are the result of time-lapse measurements at increasing frequencies of the OET. Machine learning algorithms enable automatic selection and characterization of the information content present in the OET signature so derived. Experiments are performed on three biological case studies, involving 1) the discrimination of cell types among U937 human leukemia cells, PC-3 human prostate cancer cells and HaCaT human immortalized keratinocytes; 2) the evaluation of the effects of the chemotherapeutic agent etoposide on U937 cells at different concentrations; and 3) the evaluation of the effects of different exposure times of etoposide on U937 cells. The obtained results demonstrate that multiple levels of dielectric information can be extracted via OET cell signatures and clearly pose OET as a promising tool for cell discrimination and drug response evaluation.
•We propose a novel system based on Lab-on-Chip and OET for flexible cell manipulation and in-depth single-cell analyses.•We derived spectra responses of kinematic and dynamic descriptors via TLM and OET measurements at increasing frequencies.•Machine Learning algorithms are used to characterize the information content from the OET signatures.•We proved the effectiveness in the evaluation of the effects of chemotherapy at various concentrations and exposure times.
Fabry's disease (FD) is a genetic disorder of lysosomal storage characterized by the intralysosomal accumulation of globotriaosylceramide (Gb3). This genetic mutation causes a total or partial ...deficit of the α-galactosidase (GAL) enzyme activity. FD has an incidence of 1:40000-60000 born alive. Its prevalence is higher in specific pathological conditions like chronic kidney disease (CKD). The aim of this study was to evaluate the FD prevalence in Italian renal replacement therapy (RRT) patients from Lazio region.
485 patients in RRT (hemodialysis, peritoneal dialysis, and kidney transplantation) were recruited. The screening test was performed on venous blood sample. The latter was analyzed using specific FD diagnostic kit, based on the analysis of dried blood spots on filter paper.
We found 3 cases of positivity to FD (1 female and 2 males). In addition, 1 male patient was identified with biochemical alteration indicative of GAL enzyme deficiency with a genetic variant of the GLA gene of unknown clinical significance. The FD prevalence in our population was 0.60% (1 case out 163), it rises to 0.80% (1 case out of 122) if the genetic variant of unknown clinical significance is considered. Comparing the three subpopulations, we observed a statistically significant difference in GAL activity in transplanted patients compared to dialysis patients (p<0.001).
Considering the presence of an enzyme replacement therapy able to modify FD clinical history, it is essential to try to implement FD early diagnoses. However, the screening is too expensive to be extended on large scale, due to the low prevalence of the pathology. The screening should be performed on high-risk populations.
Chronic degenerative non-communicable diseases (CDNCDs), in particular chronic kidney disease, induce gut microbiota (GM) dysbiosis, which, in turn, worsens the progression of CDNCDs and patients' ...quality of life. We analyzed literature studies to discuss the possible positive and beneficial impact of physical activity on GM composition and CV risk in CKD patients. Regular physical activity seems to be able to positively modulate the GM, reducing the systemic inflammation and consequently the production of uremic gut-derived toxins, which are directly correlated with the increase of cardiovascular risk. In particular, the accumulation of indoxyl sulphate (IS) seems to be able to induce vascular calcifications, vascular stiffness and cardiac calcifications, while p-Cresyl sulphate (p-CS) seems to be able to exert a cardiotoxic action through metabolic pathways, capable of inducing oxidative stress. In addition, trimethylamine N-oxide (TMAO) can alter lipid metabolism, inducing the production of foam cells and causing an accelerated atherosclerosis process. In this context, a regular physical activity program seems to represent an adjuvant non-pharmacological approach to the clinical management of CKD patients.
