Subtyping on Live Lymphoma Cell Lines by Raman Spectroscopy

Raman spectroscopy is a well-defined spectroscopic technique sensitive to the molecular vibrations of materials, since it provides fingerprint-like information regarding the molecular structure of the analyzed samples. It has been extensively used for non-destructive and label-free cell characterization, particularly in the qualitative and quantitative estimation of amino acids, lipids, nucleic acids, and carbohydrates. Lymphoma cell classification is a crucial task for accurate and prompt lymphoma diagnosis, prognosis, and treatment. Currently, it is mostly based on limited information and requires costly and time-consuming approaches. In this work, we are proposing a fast characterization and differentiation methodology of lymphoma cell subtypes based on Raman spectroscopy. The study was performed in the temperature range of 15–37 °C to identify the best cell measurement conditions. The proposed methodology is fast, accurate, and requires minimal sample preparation, resulting in a potentially promising, non-invasive strategy for early and accurate cell lymphoma characterization.

Elaboration and Characterization of Cu Based Nanocomposites for Electromagnetic Interferences Shielding

An electromagnetic interferences (EMI) shielding is a material that attenuates radiated electromagnetic energy. Polymer nanocomposites is a class of materials that combine electrical, thermal, dielectric, magnetic and/ or mechanical properties, which are useful for the suppression of electromagnetic interferences. In this work, we looked over the effectiveness of the electromagnetic interferences shielding of polymer-based nanocomposites. These are thin samples of epoxy resin strengthened with nanostructured Cu powders. Nanostructured Cu powders were obtained by mechanical milling using the high-energy RETSCH PM400 ball mill (200 rpm). A powder sampling was conducted after 3h, 6h, 12h, 24h, 33h, 46h and 58h milling for characterization requirements. XRD analysis via the Williamson-Hall method shows that the mean crystallites size decreases from 151.6 nm (pure Cu phase) to 13.8 nm (58 h milling). Simultaneously, the lattice strain increases from 0.1% (pure Cu phase) to 0.59% (58 h milling). The elaboration of thin samples was performed by mixing a vol./3 fractions of nanostructured Cu powder, epoxy resin and hardener. Thin slabs of 1 mm thickness were moulded for use in a rectangular wave-guide. The EMI shielding experimental involved a two ports S parameters cell measurement made of R120 metallic wave-guides of rectangular section (19.05x9.525 mm2) and operational over the frequency band of 9.84 to 15 GHz associated to a network analyser. Obtained results show moderate EMI shielding effectiveness for the milled Cu-based slabs.

THE GAIN IN SHIEDLING EFFECTIVENESS ACHIEVED BY SUPERPOSITION OF STAINLES-STEEL PLASMA COATED WOVEN FABRICS

Electromagnetic shielding based on textile fabrics is important in applications for ensuring proper work of electronic equipment and for protection of human’s health. Fibre-based materials include a good capability for a precise design of the physical and electric properties of the EM shields. There are two main methods to impart electroconductive properties to textile fabrics: insertion of conductive yarns into the fabric structure and coating with conductive layers. In our approach, both methods were applied: cotton woven fabrics with conductive yarns of stainless steel and silver, were coated by magnetron sputtering with stainless steel layers. Electromagnetic shielding effectiveness (EMSE) was determined by Transversal-Electric- Magnetic (TEM) cell measurement system, according to standard ASTM ES-07. Moreover, EMSE was determined for the superposition of the manufactured textile shields. The stainless-steel plasma coating improves EMSE with 20 dB in case of the fabrics with stainless steel yarns and with 15-17 dB in case of the fabrics with silver yarns, in the frequency range of 0.1-1000 MHz. By superposition of the plasma coated shields, the gain in EMSE achieved was of 6 dB for the fabrics with stainless steel yarns and of 5-8 dB for the fabrics with silver yarns, on the same frequency range. EMSE has significant higher values in case of the superposed shields with silver yarns and stainless-steel coating for the frequency domain of 100-1000 MHz, due to the higher thickness and the significant contribution of the multiple reflection term.

Live cell imaging of metabolic heterogeneity by quantitative fluorescent ATP indicator protein, QUEEN-37C.

Adenosine triphosphate (ATP) is often referred as the energy currency of the cell. Yet, non-invasive, real-time, and quantitative measurement of its concentration in living mammalian cells has been difficult. Here we report an improved fluorescent ATP indicator protein, QUEEN-37C, which is optimized for measuring ATP concentration in living mammalian cells. Absolute value of the ATP concentration can be estimated from the ratiometric fluorescence imaging, and its accuracy was verified by the luciferase assay. Since QUEEN-37C enables the single-cell measurement of ATP concentration, we can not only measure its mean but its distribution in the cell population, which revealed that the ATP concentration is tightly regulated in most cells. We also noted the positive correlations in the ATP concentration among adjacent cells in epithelial cell sheet and mouse embryonic stem cell colonies. Thus, QUEEN-37C would serve as a new tool for the investigation of the single cell heterogeneity of metabolic states.

Application of simultaneous time-resolved 3-D PTV and Two Colour LIF in Studying Rayleigh-Benard Convection

To study the flow topology and temperature distribution of Rayleigh-Benard convection in a highly slender cell, measurement of the simultaneous velocity and temperature in the 3-D domain is required. For this aim, implementing a simultaneous time-resolved 3-D PTV and two-colour PLIF is planned. As a part of this development, for both PTV and two-colour PLIF techniques, the experimental setup has been implemented separately to measure time-resolved 2-D velocity and temperature and is presented in this paper. For PTV, a scanning system is also utilized to scan the flow field to capture the planar velocity in different depths of the flow domain. Progress on calculation of the out-of-plane velocity component including the theory is discussed. Finally, results of the time-resolved 2-D PTV and PLIF systems are presented.

Detection of Lithium Plating in Li-Ion Cell Anodes Using Realistic Automotive Fast-Charge Profiles

The widespread use of electric vehicles is nowadays limited by the “range anxiety” of the customers. The drivers’ main concerns are related to the kilometric range of the vehicle and to the charging time. An optimized fast-charge profile can help to decrease the charging time, without degrading the cell performance and reducing the cycle life. One of the main reasons for battery capacity fade is linked to the Lithium plating phenomenon. This work investigates two methodologies, i.e., three-electrode cell measurement and internal resistance evolution during charging, for detecting the Lithium plating conditions. From this preliminary analysis, it was possible to develop new Multi-Stage Constant-Current profiles, designed to improve the performance in terms of charging time and cells capacity retention with respect to a reference profile. Four new profiles were tested and compared to a reference. The results coming from the new profiles demonstrate a simultaneous improvement in terms of charging time and cycling life, showing the reliability of the implemented methodology in preventing Lithium plating.

Microfluidic Based Physical Approaches towards Single-Cell Intracellular Delivery and Analysis

The ability to deliver foreign molecules into a single living cell with high transfection efficiency and high cell viability is of great interest in cell biology for applications in therapeutic development, diagnostics, and drug delivery towards personalized medicine. Various physical delivery methods have long demonstrated the ability to deliver cargo molecules directly to the cytoplasm or nucleus and the mechanisms underlying most of the approaches have been extensively investigated. However, most of these techniques are bulk approaches that are cell-specific and have low throughput delivery. In comparison to bulk measurements, single-cell measurement technologies can provide a better understanding of the interactions among molecules, organelles, cells, and the microenvironment, which can aid in the development of therapeutics and diagnostic tools. To elucidate distinct responses during cell genetic modification, methods to achieve transfection at the single-cell level are of great interest. In recent years, single-cell technologies have become increasingly robust and accessible, although limitations exist. This review article aims to cover various microfluidic-based physical methods for single-cell intracellular delivery such as electroporation, mechanoporation, microinjection, sonoporation, optoporation, magnetoporation, and thermoporation and their analysis. The mechanisms of various physical methods, their applications, limitations, and prospects are also elaborated.

Rituximab-Induced Hypogammaglobulinemia and Infections in AQP4 and MOG Antibody–Associated Diseases

ObjectiveTo determine the potential association between infections and rituximab (RTX)-induced hypogammaglobulinemia among patients with CNS inflammatory diseases.MethodsWe included in a prospective observational study all consecutive adults with aquaporin 4 (AQP4) or myelin oligodendrocyte glycoprotein (MOG) antibody–positive disorders treated with RTX. Dosing schedule was adapted to memory B-cell measurement.ResultsWe included 48 patients (mean age 47 [SD: 14] years; 77% females; 31 AQP4 positive and 17 MOG positive). The median follow-up was 3.6 years (range: 0.9–8.1 years). The median number of RTX infusions was 8 (range: 2–14). The median dosing interval was 6 months (range: 1.7–13.7 months). Sixty-seven symptomatic infections (SIs) were observed in 26 of 48 (54%) patients, including 13 severe infections in 9 (19%). Urinary and lower respiratory tract infections were the most frequent, representing 42% and 21% of SI. At RTX onset, the immunoglobulin G (IgG) level was abnormal in 3 of 48 (6%) patients. After RTX, 15 (31%), 11 (23%), 3 (6%), and 0 of 48 patients showed sustained IgG level <7, <6, <4, and <2 g/L, respectively. On multivariate Cox proportional hazards analysis, the main variables explaining the risk of SI were the presence of urinary tract dysfunction (hazard ratio [HR] = 34, 95% CI 4–262, p < 0.001), the dosing intervals (HR = 0.98, 95% CI 0.97–0.99, p < 0.001), and the interaction between IgG level and urinary tract dysfunction (HR = 0.67, 95% CI 0.53–0.85, p < 0.005). IgG level <6 g/L during RTX was associated with male sex (HR = 4, 95% CI 1.4–11.4, p < 0.01) and previous immunosuppression (HR = 3.4, 95% CI 1.2–10, p < 0.05).ConclusionsRTX used as maintenance therapy in CNS inflammatory diseases is frequently associated with reduced IgG level and increases the infection risk of the most vulnerable patients.