Here, we reported a study on the detection and electrical characterization of both cancer cell line and primary tumor cells. Dielectrophoresis (DEP) and electrical impedance spectroscopy (EIS) were jointly employed to enable the rapid and label-free differentiation of various cancer cells from normal ones. The primary tumor cells that were collected from two colorectal cancer patients, cancer cell lines (SW-403, Jurkat, and THP-1), and healthy peripheral blood mononuclear cells (PBMCs) were trapped first at the level of interdigitated microelectrodes with the help of dielectrophoresis. Correlation of the cells dielectric characteristics that was obtained via electrical impedance spectroscopy (EIS) allowed evident differentiation of the various types of cell. The differentiations were assigned to a “dielectric phenotype” based on their crossover frequencies. Finally, Randles equivalent circuit model was employed for highlighting the differences with regard to a series group of charge transport resistance and constant phase element for cancerous and normal cells.

Allergic rhinitis (AR) is a chronic and frequent condition characterized by an excessive response of the immune system to innocent substances encountered in the nasal mucosa. These reactions are mediated by many factors, including chemokines. Chemokine ligand 3 (CCL3, a macrophage inflammatory protein 1α) is a chemokine implicated in the activation of mast cells - white cells shown to be highly involved in orchestrating allergic reactions. The present study evaluated the role of CCL3 in AR. Thirty-nine participants, including 24 patients with AR and 15 healthy controls, were evaluated for allergies to dust mites, cat and dog danders, cockroaches (Blatella germanica), molds, grasses, weeds, and tree pollen using skin prick tests. Participants were also evaluated for inflammatory conditions by measuring total blood count with differential; concentrations of rheumatoid factor, fibrinogen, and C-reactive protein; and erythrocyte sedimentation rate. CCL3 in blood samples was measured at the Immunology Laboratory, Cantacuzino National Institute for Military Medical Research and Development, Bucharest, Romania, using Human Multianalyte Profiling Base Kits (R&D Systems Inc., Minneapolis, MN). Mean serum CCL3 concentration was significantly higher in patients with AR than in controls (15.03 ± 7.11 pg/ml vs. 8.34 ± 4.46 pg/ml, p = 0.001 [t-test] and p = 0.026 [Mann-Whitney test]). CCL3 concentrations correlated with polysensitization, defined as two or more positive prick tests per patient (r = 0.325, p = 0.046) and seasonal AR (r = 0.482, p = 0.002). Elevated levels of CCL3 were seen in our patients with AR. We have observed correlations with polysensitization and seasonal allergies. These results suggest that chemokines might play an important role in the pathogenesis of AR. In the future, chemokines might be used in endotype classification of patients with AR and as a possible target in the treatment of AR.

Protein immobilization using biopolymer scaffolds generally involves undesired protein loss of function due to denaturation, steric hindrance or improper orientation. Moreover, most methods for protein immobilization require expensive reagents and laborious procedures. This work presents the synthesis and proof of concept application of two alginate hydrogels that are able to bind proteins with polyhistidine tags by means of interaction with the crosslinking cations. Nickel (II) and cobalt (II) alginate hydrogels were prepared using a simple ionic gelation method. Hydrogels were characterized by optical microscopy and AFM, and evaluated for potential cytotoxicity. In addition, binding capacity was tested towards proteins with or without HisTAG. Hydrogels had moderate cytotoxicity and were able to exclusively bind polyhistidine-tagged proteins with a binding capacity of approximately 300 µg EGFP (enhanced green fluorescent protein) per 1 mL of hydrogel. A lyophilized hydrogel-protein complex dissolved upon the addition of PBS and allowed the protein release and regain of biological activity. In conclusion, the nickel (II) and cobalt (II) alginate biopolymers provided an excellent platform for the “carry and release” of polyhistidine-tagged proteins.

This study proposes a feasible approach for the rapid, sensitive, and label-free identification of cancerous cells based on dielectrophoretic (DEP) manipulation and electrical characterization. In this method, the concentration of target cells at the level of customized microelectrodes via DEP is first determined, followed by an electrical impedance evaluation. The study demonstrates the capacity of the methodology to electrically differentiate HT-29 cancer cells from healthy blood cells based on their impedance spectra. Within a higher frequency domain, the electrical impedance of trapped cancer cells was significantly lower compared with the normal ones. In order to evaluate the functionality and reproducibility of the proposed method, the influence of the DEP and EIS (electrical impedance spectroscopy) operating voltages on the electrical characterization of trapped HT-29 cells was analyzed.

Influenza virus infections represent an ongoing public health threat as well as an economic burden. Although seasonal influenza vaccines have been available for some decades, efforts are being made to generate new efficient, flexible, and cost-effective technologies to be transferred into production. Our work describes the development of a model influenza hemagglutinin antigen that is capable of inducing protection against viral challenge in mice. High amounts of the H1 hemagglutinin ectodomain, HA_18–528, were expressed in a bacterial system as insoluble inclusion bodies. Solubilization was followed by a thorough differential scanning fluorimetry (DSF)-guided optimization of refolding, which allows for fast and reliable screening of several refolding conditions, yielding tens of milligrams/L of folded protein. Structural and functional analysis revealed native-like folding as well as the presence of a mix of monomers and oligomers in solution. Mice immunized with HA_18–528 were protected when exposed to influenza A virus as opposed to mice that received full-length denatured protein. Sera of mice immunized with HA_18–528 showed both high titers of antigen-specific IgG1 and IgG2a isotypes as well as viral neutralization activity. These results prove the feasibility of the recombinant bacterial expression system coupled with DSF-guided refolding in providing influenza hemagglutinin for vaccine development.

Peri-implantitis (PI) is a current concern whose understanding and resolution are ongoing. We aimed to evaluate in vivo a new treatment with antibacterial properties, based on bacterial lysates obtained from the strains of Aggregatibacter actinomycetemcomitans, Streptococcus oralis, and Fusobacterium nucleatum. This research was conducted on 30 rats with PI which were divided into three groups and treated with antibiotic and anti-inflammatory (AAi) drugs, bacterial lysates (BLs), and saline (C), respectively. The monitoring period included the clinical and paraclinical examination where hematological, immunological, imaging, and histopathological analysis were performed. No particular clinical signs were observed, but the radiological examination showed the loss of all implants in group C, in contrast to group BL which had the highest survival rate of devices. White cells showed a decrease from the PI period, as did the immunological analysis. Only IL-6 showed an increase in the AAi and BL groups. Histopathologically, the C group presented a high degree of bone destruction, and in the BL group, many attenuated inflammatory phenomena appeared compared to the AAi animals. Bacterial lysates have similar effects to antibiotic-based therapeutic regimens for PI, and their future use may help to improve the current therapeutic management of the disease.