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Iron(iii) oxide nanoparticles doped with magnesium exposed to an alternating magnetic field induced cytotoxic effects on lung cancer cells (A549).

Studies over the last decade have shown that matrix metalloproteinases (MMPs) play a key role in the growth and metastasis of cancer. This zinc-dependent family of endopeptidases is crucial for the degradation of extracellular matrix (ECM), as well as serves as important ECM transducers which have been recognized as early biomarkers for both cancer diagnosis and treatment. In this study, we designed a new type of voltammetric biosensor, composed of a glycine-methionine dipeptide conjugated covalently to ferrocene (Gly-Met-Fc), for fast and ultrasensitive detection of the active form of MMP-9 in plasma samples. The detection was based on specific enzymatic cleavage of the Gly-Met peptide bond, which was monitored by voltammetry and gravimetry measurements. The ferrocene units act as voltammetric visualizers for the detection process. The cysteamine layer directly anchored to the gold surface ensured that the packing density of Gly-Met-Fc in the receptor layer was appropriate for the sensitive detection of MMP-9 in its active form. The developed biosensor was characterized by the widest analytical range (2.0·10−6 - 5.0 μg⋅mL−1) and low detection limit (0.04 pg⋅mL−1). Another valuable feature of the proposed biosensor is that it can be applied directly to the plasma samples without any additional preparation step and thus speeds up the analysis.

Nearly half of patients with advanced and metastatic melanomas harbor a BRAF mutation. Vemurafenib (VEM), a BRAF inhibitor, is used to treat such patients, however, responses to VEM are very short-lived due to intrinsic, adaptive and/or acquired resistance. In this context, we present the action of the B-Raf serine-threonine protein kinase inhibitor (vemurafenib) on the glycans structure and metallomics profiles in melanoma cells without (MeWo) and with (G-361) BRAF mutations. The studies were performed using &alpha, 1-acid glycoprotein (AGP), a well-known acute-phase protein, and concanavalin A (Con A), which served as the model receptor. The detection of changes in the structure of glycans can be successfully carried out based on the frequency shifts and the charge transfer resistance after interaction of AGP with Con A in different VEM treatments using QCM-D and EIS measurements. These changes were also proved based on the cell ultrastructure examined by TEM and SEM. The LA-ICP-MS studies provided details on the metallomics profile in melanoma cells treated with and without VEM. The studies evidence that vemurafenib modifies the glycans structures and metallomics profile in melanoma cells harboring BRAF mutation that can be further implied in the resistance phenomenon. Therefore, our data opens a new avenue for further studies in the short-term addressing novel targets that hopefully can be used to improve the therapeutic regiment in advanced melanoma patients. The innovating potential of this study is fully credible and has a real impact on the global patient society suffering from advanced and metastatic melanomas.

This work presents a new look at the application of cyclodextrins (CD) as a drug nanocarrier. Two different cyclodextrins (&alpha, CD, &beta, CD) were covalently conjugated to branched polyethylenimine (PEI), which was additionally functionalized with folic acid (PEI-&beta, CD-&alpha, CD-FA). Here, we demonstrated that the combination of &alpha, CD and &beta, CD enabled to load and control release of two anticancer drugs: doxorubicin (DOX) and beta-lapachone (beta-LP) (DOX in &beta, CD and beta-LP into &alpha, CD) via host-guest inclusion. The PEI-&beta, CD(DOX)-&alpha, CD-FA nanoconjugate was used to transport anticancer drugs into A549 lung cancer cells for estimation the cytotoxic and antitumor effect of this nanoconjugate. The presence of FA molecules should facilitate the penetration of studied nanoconjugate into the cell. Whereas, the non-cellular experiments proved that the drugs are released from the carrier mainly in the pH 4.0. The release mechanism is found to be anomalous in all studied cases.

Fast, simple in use and highly effective voltammetric enantiosensor dedicated for determination of thalidomide (TD) enantiomers (especially towards the toxic (S)-enantiomer) in blood plasma is still desirable. Here we have proven that newly synthesized chiral naphthalene diimide (NDI) derivatives are excellent electroactive materials for TD enantiosensors. The recognition process relies on the specific interaction between the chiral NDI receptor and the thalidomide enantiomer of the opposite configuration. This unique specific interaction between (S)-thalidomide and (R)-NDI derivative counterparts, evident in the DPV voltammograms, was confirmed by molecular modeling. The demonstrated voltammetric enantiosensors are characterized by the low detection limit at the level of μg·L−1, wide analytical range from 5·10−4 – 10 mg·L−1, high selectivity and long lifetime. The results of the recovery rates showed a very good degree of accuracy towards the determination of (S)-thalidomide in the blood samples, so it can be successfully used in the analysis of clinical samples.

Simple, selective and sensitive analytical devices are of a great importance for medical application. Herein, we developed highly selective immunosensor for electrochemical detection of C-reactive protein (CRP) in blood sample. Branched polyethylenimine functionalized with ferrocene residues (PEI-Fc) was the main element of the recognition layer, which allowed: (i) covalent binding of an antibody in its most favorable orientation and (ii) voltammetric detection of the C-reactive protein. Anchoring of PEI-Fc to the electrode surface through the electrodeposition process leads to the formation of thin, stable and reproducible layers, which is extremely important in the case of electrochemical immunosensing. The proposed analytical device is characterized by high selectivity and sensitivity and can be successfully used in the concentration range of CRP from 1 to 5·104 ng mL−1. The determined limit of detection was circa 0.5 and 2.5 ng mL−1 for voltammetric and impedance analysis, respectively. The developed analytical device has also been successfully applied for the analysis of CRP level in rat blood samples.

An effective, fast, facile and direct electrochemical method of determination of hemoglobin (Hb) in blood sample without any sample preparation is described. The method is accomplished by using the ferromagnetic electrode modifier (carbon-encapsulated iron nanoparticles) and an external magnetic field. The successful voltammetric determination of hemoglobin is achieved in PBS buffer as well as in the whole blood sample. The obtained results show the excellent electroactivity of Hb. The measurements are of high sensitivity and good reproducibility. The detection limit is estimated to be 0.7 pM. The electrochemical determination data were compared with the gravimetric data obtained with a quartz crystal microbalance. The agreement between these results is very good. The changes of the electrode surface morphology before and after Hb detection are monitored by electron microscopy. The functionality of the electrochemical sensor is tested with human and rat blood samples. The concentration of hemoglobin in the blood samples determined by using voltammetric/gravimetric detection is in perfect agreement with the data obtained from typical clinical analysis.

The way of immobilization of the monoclonal antibody (type IgG) on the electrode surface has a significant effect on the amount of the immobilized protein and in consequence on current signal of protein. Herein, we demonstrate that the application of appropriately functionalized phenyl film allowed us to control the orientation of the antibody (Ab) molecules on the electrode surface. The influence of Ab orientation on the efficiency of antigen-antibody interaction was tested with an example blood plasma protein (ferritin; Ft). To control the orientation of Ab molecules the phenyl films containing –COOH or –NH2 groups were applied. Contrary to aminoethylophenyl layer, the carboxyphenyl film guaranteed the shortest distance between the redox center of the protein and the electrode surface. Additionally, the application of an external magnetic field together with magnetic nanoparticles allowed achieving the best orientation to observe well-defined ferritin current signals. The proposed method of ferritin detection can be successfully used in the concentration range of Ft between 0.1 and 30 µg dL−1. The detection limit for a carboxyphenyl film was estimated as 0.40 ± 0.04 and 0.13 ± 0.04 µg dL−1 for impedance and voltammetric measurements, respectively. In turn, for an aminoethylophenyl film the detection limit was 0.03 ± 0.002 (electrochemical impedance spectroscopy; EIS) and 0.02 ± 0.002 µg dL−1 (differential pulse voltammetry, DPV). The interday precision (reproducibility) was calculated (4.10 ÷ 9.10% RSD) together with the intraday precision / repeatability (3.20 ÷ 8.0% RSD) for the studied samples. The functionality of the sensor has been tested on rat blood samples. Based on the performed investigations it can be stated that the developed sensor was characterized by high selectivity and good sensitivity.

Worldwide, drug-induced liver injury (DILI) is a major cause of hepatic failure. It is also the leading cause of withdrawal, cautionary labeling, and restricted usage of licensed drugs; therefore, European Medicines Agency (EMA) and United States Food and Drug Administration (FDA) warn that the existing methods of assessing DILI are insufficient and that some of the translational biomarkers of hepatotoxicity must be relooked. Magnetic resonance imaging (MRI) seems to be a proper tool in elucidating the effects of DILI in both preclinical and clinical studies, providing excellent visualization of the morphology of the liver parenchyma. Therefore, herein, we propose preclinical MRI assessment of liver injury in experimental paracetamol-treated rats. Quantitative MRI clearly provides evidence of adverse effects in the liver tissue caused by a single overdose of paracetamol (1 g kg−1 and 1.5 g kg−1 b.w.). The results of the MRI were confirmed by the histopathological examination (HE it has a potential to serve as the early diagnostic tool for the prediction of DILI in preclinical evaluation.

Objectives The purpose of this study was to assess the value of contrast-enhanced magnetic resonance cholangiography (MRC) performed in different time delays after injection of gadoxetic acid disodium (Gd-EOB-DTPA) for the diagnosis of active bile leak. Methods This retrospective analysis included Gd-EOB-DTPA enhanced MR images of 34 patients suspected of bile leak. Images were acquired 20–25 min after Gd-EOB-DTPA injection. If there was inadequate contrast in the bile ducts then delayed images after 60–90 min and 150–180 min were obtained. Results were correlated with intraoperative findings, ERCP results, clinical data, laboratory tests, and follow-up examinations. Results Gd-EOB-DTPA enhanced MRC yielded an overall sensitivity of 96.4%, specificity of 100% and accuracy of 97.1% for the diagnosis of an active bile leak. The sensitivity of 20–25 min delayed MR images was 42.9%, of combined 20–25 min and 60–90 min delayed images was 92.9% and of combined 20–25 min, 60–90 min and 150–180 min delayed images was 96.4%. Conclusions Gd-EOB-DTPA enhanced MRC utilizing delayed phase images was effective for detecting the presence and location of active bile leaks. The images acquired 60–180 min post-injection enabled identification of bile leaks even in patients with a dilated biliary system or moderate liver dysfunction.

Carbon-encapsulated iron nanoparticles (CEINs) qualified as metal-inorganic hybrid nanomaterials offer a potential scope for an increasing number of biomedical applications. In this study, we have focused on the investigation of cellular fate and resulting cytotoxic effects of CEINs synthesized using a carbon arc route and studied in murine endothelial (HECa-10) cells. The CEIN samples were characterized as pristine (the mean diameter between 47 and 56nm) and hydrodynamic (the mean diameter between 270 and 460nm) forms and tested using a battery of methods to determine the cell internalization extent and cytotoxicity effects upon to the exposures (0.0001-100μg/ml) in HECa-10 cells. Our studies evidenced that the incubation with CEINs for 24h is accompanied with substantial changes of Zeta potential in cells which can be considered as a key factor for affecting the membrane transport, cellular distribution and cytotoxicity of these nanoparticles. The results demonstrate that CEINs have entered the endothelial cell through the endocytic pathway rather than by passive diffusion and they were mainly loaded as agglomerates on the cell membrane and throughout the cytoplasm, mitochondria and nucleus. The studies show that CEINs induce the mitochondrial and cell membrane cytotoxicities in a dose-dependent manner resulting from the internal dosages due to CEIN agglomerates. Our results highlight the importance of the physicochemical characterization of CEINs in studying the magnetic nanoparticle-endothelial cell interactions because the CEIN mass agglomerates can sediment more or less rapidly in culture models.

Background The aim of this study was to determine if the appearance of hepatic epithelioid hemangioendothelioma (HEHE) on state-of-the-art MRI including hepatocyte phase after administration of hepatobiliary contrast agent can facilitate preoperative diagnosis and identification of potential candidates for liver transplantation. Material and methods The study group comprised 6 patients with pathologically confirmed HEHE. Analysis included signal characteristics of 55 tumor nodules (maximum of 10 lesions per patient) on T2-weighted images, dynamic contrast-enhanced, 5-minute delayed, and hepatobiliary phase images. Results The most common feature of HEHE, observed in 84% of lesions, was progressive contrast-enhancement, followed by subcapsular location (66%), confluent appearance (60%) and hyper- or isointensity on hepatobiliary phase images (53%). In 5 of 6 patients, capsular retraction was observed. Conclusions The appearance of HEHE on hepatobiliary phase images was variable, but examined tumors often demonstrated hyper- or isointensity, most probably due to prolonged retention of contrast material. These features, along with typical morphology (subcapsular, confluent nodules, with progressive enhancement and capsular retraction), may contribute to correct diagnosis and recognition of potential candidates for liver transplantation.

Carbon-encapsulated iron nanoparticles (CEINs) have recently emerged as a new class of magnetic nanomaterials with a great potential for an increasing number of biomedical applications. To address the current deficient knowledge of cellular responses due to CEIN exposures, we focused on the investigation of internalization profile and resulting cytotoxic effects of CEINs (0.0001–100 μg/ml) in murine glioma cells (GL261) in vitro . The studied CEIN samples were characterized (TEM, FT-IR, Zeta potential, Boehm titration) and examined as raw and purified nanomaterials with various surface chemistry composition. Of the four type CEINs (the mean diameter 47–56 nm) studied here, the as-synthesized raw nanoparticles (Fe@C/Fe) exhibited high cytotoxic effects on the plasma cell membrane (LDH, Calcein AM/PI) and mitochondria (MTT, JC-1) causing some pro-apoptotic evens (Annexin V/PI) in glioma cells. The effects of the purified (Fe@C) and surface-modified (Fe@C-COOH and Fe@C-(CH 2 ) 2 COOH) CEINs were found in quite similar patterns; however, most of these cytotoxic events were slightly diminished compared to those induced by Fe@C/Fe. The study showed that the surface-functionalized CEINs affected the cell cycle progression in both S and G2/M phases to a greater extent compared to that of the rest of nanoparticles studied to data. Taken all together, the present results highlight the importance of the rational design of CEINs as their physicochemical features such as morphology, hydrodynamic size, impurity profiles, and especially surface characteristics are critical determinants of different cytotoxic responses.

A chemical property space defines the adaptability of a molecule to changing conditions and its interaction with other molecular systems determining a pharmacological response. Within a congeneric molecular series (compounds with the same derivatization algorithm and thus the same brute formula) the chemical properties vary in a monotonic manner, i.e., congeneric compounds share the same chemical property space. The chemical property space is a key component in molecular design, where some building blocks are functionalized, i.e., derivatized, and eventually self-assembled in more complex systems, such as enzyme-ligand systems, of which (physico-chemical) properties/bioactivity may be predicted by QSPR/QSAR (quantitative structure-property/activity relationship) studies. The system structure is determined by the binding type (temporal/permanent; electrostatic/covalent) and is reflected in its local electronic (and/or magnetic) properties. Such nano-systems play the role of molecular devices, important in nano-medicine. In the present article, the behavior of polyethylenimine (PEI) macromolecules (linear LPEI and branched BPEI, respectively) with respect to the glucose oxidase enzyme GOx is described in terms of their (interacting) energy, geometry and topology, in an attempt to find the best shape and size of PEIs to be useful for a chosen (nanochemistry) purpose.

To compare the efficacy of two quantitative methods for discrimination between benign and malignant focal liver lesions (FLLs): apparent diffusion coefficient (ADC) values and T2 relaxation times.Seventy-three patients with 215 confirmed FLLs (115 benign, 100 malignant) underwent 1.5-T MRI with respiratory-triggered single-shot SE DWI (b = 50, 400, 800) and dual-echo T2TSE (TR = 3,000 ms; TE1 = 84 ms; TE2 = 228 ms). ADC values and T2 relaxation times of FLLs were calculated. Sensitivity, specificity and accuracy of both techniques in diagnosing malignancy were assessed.The mean ADC value of malignant tumours (1.07 × 10(-3) mm(2)/s) was significantly lower (P 0.05) than that of benign lesions (1.86 × 10(-3) mm(2)/s ); however, with the use of the optimal cut-off value of 1.25 × 10(-3) mm(2)/s, 20 false positive (FP) and 20 false negative (FN) diagnoses of malignancy were noted, generating 79 % sensitivity, 82.6 % specificity and 80.9 % accuracy. The mean T2 relaxation time of malignant tumours (64.4 ms) was significantly lower (P 0.05) than that of benign lesions (476.1 ms). At the threshold of 107 ms 22 FP and 1 FN diagnoses were noted; the sensitivity was 99 %, specificity 80.9 % and accuracy 89.3 %.Quantitative analysis of T2 relaxation times yielded significantly higher sensitivity and accuracy in diagnosing malignant liver tumour than ADC values.• Diffusion-weighted magnetic resonance imaging is increasingly used for liver lesions. • But ADC values demonstrated only moderate accuracy for differentiation of liver lesions. • T2 relaxation times yielded higher accuracy in diagnosing malignant liver tumours. • Both ADC and T2 values overlapped between focal nodular hyperplasia and malignant lesions. • Nevertheless T2 liver mapping could be valuable for evaluating focal liver lesions.

Glucose oxidase (GOx) is an enzyme produced by Aspergillus, Penicillium and other fungi species. It catalyzes the oxidation of β-d-glucose (by the molecular oxygen or other molecules, like quinones, in a higher oxidation state) to form d-glucono-1,5-lactone, which hydrolyses spontaneously to produce gluconic acid. A coproduct of this enzymatic reaction is hydrogen peroxide (H2O2). GOx has found several commercial applications in chemical and pharmaceutical industries including novel biosensors that use the immobilized enzyme on different nanomaterials and/or polymers such as polyethylenimine (PEI). The problem of GOx immobilization on PEI is retaining the enzyme native activity despite its immobilization onto the polymer surface. Therefore, the molecular dynamic (MD) study of the PEI ligand (C14N8_07_B22) and the GOx enzyme (3QVR) was performed to examine the final complex PEI-GOx stabilization and the affinity of the PEI ligand to the docking sites of the GOx enzyme. The docking procedure showed two places/regions of major interaction of the protein with the polymer PEI: (LIG1) of −5.8 kcal/mol and (LIG2) of −4.5 kcal/mol located inside the enzyme and on its surface, respectively. The values of enthalpy for the PEI-enzyme complex, located inside of the protein (LIG1) and on its surface (LIG2) were computed. Docking also discovered domains of the GOx protein that exhibit no interactions with the ligand or have even repulsive characteristics. The structural data clearly indicate some differences in the ligand PEI behavior bound at the two places/regions of glucose oxidase.