Dr. Daniel CRISAN

Paper-based devices hold great promise in biosensing, but the choice of electrode materials influences performance. Here, we report a paper-based electrochemical sensor developed for nucleic acid quantification, in a sandwich-type architecture integrating 3-electrode systems on metallized electrospun polymeric fibers. A 3D-printed hydrophobic barrier on the chromatographic paper defines injection and testing zones. Fluid diffusion through paper and concentration gradients are considered in the design. Electrochemical characterization is performed using 40 mu L of methylene blue solution, which interacts with double-stranded nucleic acids, reducing its redox activity. This interaction mechanism within the paper substrate is confirmed by spectroscopy. The sensor achieves detection of nucleic acids in 3 min with 2 mu L of solution. Real sample analysis is performed for the quantification of PCR-amplified genes with a limit of detection of 1.38 ng mu L-1. The device serves as a promising point-of-care diagnostics tool for the direct quantification of amplified genetic material.

Over the past decades, extensive research has demonstrated and confirmed the antioxidant potency of polyphenols, which due to these properties, are now widely recognized for their ability to enhance cell viability. This study investigates the antioxidant capacity of seven green leafy vegetables on in vitro mouse (L929) and human (BJ) fibroblast cell lines, aiming to determine if polyphenols could aid the regeneration of connective tissue, which would be strongly beneficial in wound healing and tissue repair, a process where fibroblast thriving is essential. The antioxidant capacity of extracts was investigated using the 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay, UV-Vis spectroscopy, and amperometry, with quercetin serving as a standard. The EC50 values for the extracts were equivalent to approximately 5 mu M quercetin, derived from a concentration range of 20 to 100 mg of fresh leaves per mL. In vitro investigations revealed that all extracts, except lovage, promoted high viability (over 80%) in both cell cultures, as shown by MTS assay results and fluorescence microscopy. Additionally, Tumor Necrosis Factor (TNF)-alpha and Lipopolysaccharide (LPS) were employed as reactive oxygen species (ROS) antagonists, and the Dichloro-dihydro-fluorescein diacetate (DCFH-DA) assay was used to demonstrate the extract's scavenging activity on fibroblasts in vitro. For some extracts, a reduction in oxidative stress compared to the cells basal metabolic conditions was observed.

This work describes the development of a flexible uric acid (UA) biosensor based on palladium-coated submicrometer electrospun poly(methylmethacrylate) (PMMA) fibers metalized with gold and attached to polyethylene terephthalate substrate (Pd/Au/PMMA/PET). The morphological characterization conducted by scanning electron microscopy revealed nanoscale Pd dendritic structures. Electrochemical investigations in the absence and in the presence of redox probes demonstrated that these Pd nanostructures are responsible for a six-fold increase in the electroactive area and enhanced electron transfer kinetics when compared to the gold-coated electrospun fibers. The UA biosensor obtained by immobilizing the uricase enzyme (UrOx) onto the Pd/Au/PMMA/PET electrode surface, allowed UA detection with a sensitivity of 431 mu A cm(-2) mM(-1) and a limit of detection of 12 mu M. Investigation of the redox reactions of hydrogen peroxide (a product of the enzymatic oxidation of UA by UrOx) at the Pd/Au/PMMA/PET electrode demonstrated that the working principle of the biosensor is based on the reduction of PdO produced at the electrode surface during the spontaneous reduction of hydrogen peroxide on Pd. This allows a biosensor operating potential of -0.05 V (vs Ag/AgCl) with high selectivity. The UrOx/Pd/Au/PMMA/PET biosensor was applied for UA detection in body fluids (sweat, urine, and blood serum) with recovery values between 98 and 105%, which were validated by high-performance liquid chromatography analysis. The stability of the device was evaluated over a period of 3 months, retaining 78% of the initial sensitivity, and reproducibility with RSD = 4.9% was achieved. The analytical performance of the biosensor under harsh mechanical deformations and at physiological temperatures demonstrated the potential applications of the device to wearable sensing platforms.

Among many iron-based superconductors, isovalently substituted BaFe2(As1-xPx)2 displays, for x approximate to 0.3, apart from the quite usual Second Magnetization Peak (SMP) in the field dependence of the critical current density, an unusual peak effect in the temperature dependence of the critical current density in the constant field, which is related to the rhombic-to-square (RST) structural transition of the Bragg vortex glass (BVG). By using multi-harmonic AC susceptibility investigations in three different cooling regimes-field cooling, zero-field cooling, and field cooling with measurements during warming up-we have discovered the existence of a temperature region in which there is a pronounced magnetic memory effect, which we attributed to the direction of the structural transition. The observed huge differences in the third harmonic susceptibility at low and high AC frequencies indicates the difference in the time-scale of the structural transition in comparison with the timescale of the vortex excitations. Our findings show that the RST influence on the vortex dynamics goes beyond the previously observed influence on the onset of the SMP.

For practical applications of superconductors, understanding the vortex matter and dynamics is of paramount importance. An important issue in this context is the transition of the vortex glass, which is a true superconducting phase, into a vortex liquid phase having a linear dissipation. By using multi-harmonic susceptibility studies, we have investigated the vortex glass-vortex liquid phase transitions in CaKFe4As4 and BaFe2(As0.68P0.32)(2) single crystals. The principle of our method relates the on-set of the third-harmonic susceptibility response with the appearance of a vortex-glass phase in which the dissipation is non-linear. Similar to the high-critical temperature cuprate superconductors, we have shown that even in these iron-based superconductors with significant lower critical temperatures, such phase transition can be treated as a melting in the sense of Lindemann's approach, considering an anisotropic Ginzburg-Landau model. The experimental data are consistent with a temperature-dependent London penetration depth given by a 3D XY fluctuations model. The fitting parameters allowed us to extrapolate the vortex melting lines down to the temperature of liquid hydrogen, and such extrapolation showed that CaKFe4As4 is a very promising superconducting material for high field applications in liquid hydrogen, with a melting field at 20 K of the order of 100 T.

Among various "families" of iron-based superconductors, the quite recently discovered AeAFe(4)As(4) (where Ae is an alkali-earth metal and A is an alkali metal) has high critical current density, a very high upper critical field, and a low anisotropy, and has recently received much interest for the possibility of high magnetic field applications at the liquid hydrogen temperature. We have performed DC magnetization relaxation and frequency-dependent AC susceptibility measurements on high-quality single crystals of CaKFe4As4 with the aim of determining the pinning potential U*. The temperature dependence of U* displays a clear crossover between elastic creep and plastic creep. At temperatures around 27-28 K, U* has a very high value, up to 1200 K, resulting in an infinitesimally small probability of thermally activated flux jumps. From the dependence of the normalized pinning potential on irreversible magnetization, we have determined the creep exponents in the two creep regimes, which are in complete agreement with theoretical models. The estimation of the pinning potential from multifrequency AC susceptibility measurements was possible only near the critical temperature due to equipment limitations, and the resulting value is very close to the one that resulted from the magnetization relaxation data. Magnetic hysteresis loops revealed a second magnetization peak and very high values of the critical current density.

For high-field power applications of high-temperature superconductors, it became obvious in recent years that nano-engineered artificial pinning centers are needed for increasing the critical current and pinning potential. As opposed to the artificial pinning centers obtained by irradiation with various particles, which is a quite expensive approach, we have studied superconducting samples having self-assembled defects, created during the sample fabrication, that act as effective pinning centers. We introduced a simple, straight-forward method of estimating the frequency-dependent critical current density by using frequency-dependent AC susceptibility measurements, in fixed temperatures and DC magnetic fields, from the positions of the maxima in the dependence of the out-of-phase susceptibility on the amplitude of AC excitation magnetic field. The results are compatible with a model that stipulates a logarithmic dependence of the pinning potential on the probing current. A mathematical derivation allowed us to estimate from the experimental data the pinning potentials in various samples, and in various DC magnetic fields. The resulted values indicate large pinning potentials, leading to very small probability of magnetic flux escaping the pinning wells, hence, leading to very high critical currents in high magnetic fields.

The use of gold nanoparticles/superoxide dismutase (AuNP/SOD) bioconjugates is described as building blocks in SOD biosensor development for the quantification of superoxide in cell culture media. AuNP functionalization with 11-mercaptoundecanoic acid (MUA) and 4-mercaptobenzoic acid (MBA) (AuNPMUA and AuNPMBA) was used to improve SOD immobilization through EDC/NHS coupling using their -COOH terminus, leading to the formation of more stable bioconjugates. AuNP and AuNP/SOD bioconjugates were characterized by SEM to determine their size and morphology, UV-Vis for optical properties, FT-IR, and Raman spectroscopies for chemical functional group analysis and EDX for elemental analysis. Electrochemical methods were used to characterize the Au/AuNP-modified electrodes. For the optimization of the biosensor architecture, different AuNP/enzyme bioconjugates were prepared by varying the amount of both enzyme and AuNP, as well as their incubation time. Finally, the biosensors incorporating the bioconjugates were characterized by fixed potential amperometry and voltammetric analysis in order to establish the enzymatic mechanism and to elucidate the best biosensor architecture for monitoring superoxide in cell culture media. The best sensitivity value for superoxide detection corresponded to 41.2 nA mu M cm(-2), achieved by a biosensor based on AuNPMBA/SOD bioconjugates monitored through fixed potential amperometry at 0.3 V vs. Ag/AgCl, with a limit of detection of 1.0 mu M, and overall very good operational stability, maintaining 91% of the initial sensitivity after 30 days. Finally, the optimized biosensor was employed for the quantification of successive additions of superoxide in cell culture media, with excellent recovery values.

We have investigated the pinning potential of high-quality single crystals of superconducting material CaKFe4As4 having high critical current density and very high upper critical field using both magnetization relaxation measurements and frequency-dependent AC susceptibility. Preliminary studies of the superconducting transition and of the isothermal magnetization loops confirmed the high quality of the samples, while temperature dependence of the AC susceptibility in high magnetic fields show absolutely no dependence on the cooling conditions, hence, no magnetic history. From magnetization relaxation measurements were extracted the values of the normalized pinning potential U*, which reveals a clear crossover between elastic creep and plastic creep. The extremely high values of U*, up to 1200 K around the temperature of 20 K lead to a nearly zero value of the probability of thermally-activated flux jumps at temperatures of interest for high-field applications. The values of the creep exponents in the two creep regimes resulted from the analysis of the magnetization relaxation data are in complete agreement with theoretical models. Pinning potentials were also estimated, near the critical temperature, from AC susceptibility measurements, their values being close to those resulted (at the same temperature and DC field) from the magnetization relaxation data.

The molecular structure of the 8-hydroxyquinoline-bis (2-phenylpyridyl) iridium (IrQ(ppy)(2)) dual emitter organometallic compound is determined based on detailed 1D and 2D nuclear magnetic resonance (NMR), to identify metal-ligands coordination, isomerization and chemical yield of the desired compound. Meanwhile, the extended X-ray absorption fine structure (EXAFS) was used to determine the interatomic distances around the iridium ion. From the NMR results, this compound IrQ(ppy)(2) exhibits a trans isomerization with a distribution of coordinated N-atoms in a similar way to facial Ir(ppy)(3). The EXAFS measurements confirm the structural model of the IrQ(ppy)(2) compound where the oxygen atoms from the quinoline ligands induce the splitting of the next-nearest neighboring C in the second shell of the Ir3+ ions. The high-performance liquid chromatography (HPLC), as a part of the detailed molecular analysis, confirms the purity of the desired IrQ(ppy)(2) organometallic compound as being more than 95%, together with the progress of the chemical reactions towards the final compound. The theoretical model of the IrQ(ppy)(2), concerning the expected bond lengths, is compared with the structural model from the EXAFS and XRD measurements.

The recent advances in genetic engineering demand the development of conceptually new methods to prepare and identify efficient vectors for the intracellular delivery of different nucleotide payloads ranging from short single-stranded oligonucleotides to larger plasmid double-stranded circular DNAs. Although many challenges still have to be overcome, polymers hold great potential for intracellular nucleotide delivery and gene therapy. We here develop and apply the postpolymerization modification of polyhydrazide scaffolds, with different degree of polymerization, for the preparation of amphiphilic polymeric vehicles for the intracellular delivery of a circular plasmid DNA. The hydrazone formation reactions with a mixture of cationic and hydrophobic aldehydes proceed in physiologically compatible aqueous conditions, and the resulting amphiphilic polyhydrazones are directly combined with the biological cargo without any purification step. This methodology allowed the preparation of stable polyplexes with a suitable size and zeta potential to achieve an efficient encapsulation and intracellular delivery of the DNA cargo. Simple formulations that performed with efficiencies and cell viabilities comparable to the current gold standard were identified. Furthermore, the internalization mechanism was studied via internalization experiments in the presence of endocytic inhibitors and fluorescence microscopy. The results reported here confirmed that the polyhydrazone functionalization is a suitable strategy for the screening and identification of customized polymeric vehicles for the delivery of different nucleotide cargos.

Among the great number of sol-gel prepared nanomaterials, TiO2 has attracted significant interest due to its high photocatalytic activity, excellent functionality, thermal stability and non-toxicity. The photocatalytic degradation of pollutants using un-doped and doped TiO2 nanopowders or thin films is very attractive for applications in environmental protection, as a possible solution for water purification. The present work describes a comparative structural and chemical study of un-doped TiO2 and the corresponding S- and Ag-doped materials. The photocatalytic activity was established by testing the degradation of organic chloride compounds from aqueous solutions. Sol-gel Ag-doped TiO2 coatings, prepared by co-gelation and sol-gel Ag-doped TiO2 coatings obtained from nanopowders were also compared. Their structural evolution and crystallization behaviour (lattice parameters, crystallite sizes, internal strains) with thermal treatment were followed by thermal analysis, X-ray diffraction, transmission electron microscopy, atomic force microscopy and specific surface areas measurements. X-ray photoelectron spectroscopy analyses were performed to characterize the surface composition and S or Ag speciation, which was used to interpret the catalytic data.

Among the great number of sol-gel materials prepared, TiO2 holds one of the most important places due to its photocatalytic properties, both in the case of powders and coatings. Impurity doping is one of the typical approaches to extend the spectral response of a wide band gap semiconductor to visible light. This work has studied some un-doped and Pd-doped sol-gel TiO2 nanopowders, presenting various surface morphologies and structures. The obtained powders have been embedded in vitreous TiO2 matrices and the corresponding coatings have been prepared by dipping procedure, on glass substrates. The relationship between the synthesis conditions and the properties of titania narrosized materials, such as thermal stability, phase composition, crystallinity, morphology and size of particles, and the influence of dopant was investigated. The influence of Pd on TiO2 crystallization both for supported and unsupported materials was studied (lattice parameters, crystallite sizes, internal strains). The hydrophilic properties of the films were also connected with their structure, composition and surface morphology. The methods used for the characterization of the materials have been: simultaneous thermogravimetry and differential thermal analysis, powder X-ray diffraction, electron microscopy (TEM, SAED) and AFM.

Pure TiO2 and S-doped TiO2 sol-gel nanopowders were prepared by controlled hydrolysis-condensation of titanium alkoxides. The influence of different Ti-alkoxides (tetraethyl-, tetraisopropyl- and tetrabutyl-orthotitanate) used in obtaining TiO2 porous materials in similar conditions (water/alkoxide ratio, solvent/alkoxide ratio, pH and temperature of reaction) has been investigated. The relationship between the synthesis conditions and the properties of titania nanosized powders, such as thermal stability, phase composition, crystallinity, morphology and size of particles, BET surface area and the influence of dopant was investigated. The nature of the alkyl group strongly influences the main characteristics of the obtained oxide powders, fact which is pointed out by thermal analysis, X-ray diffraction, TEM and BET surface area measurements.

The use of IR spectroscopic method to choose the best batches of ZSM-5 zeolites. is discussed. The crystallinity and the particle size were estimated from IR spectra and by XRD and electron microscopy measurements.