This study presents the development of an advanced drug delivery system based on the modification of surface properties of mesoporous silica for colorectal cancer therapy. Glycine-modified mesoporous silica nanoparticles (MCM-Gly) were obtained in a unique two-step synthesis approach, based on the condensation reaction between (3-aminopropyl)triethoxysilane and glycine without the use of coupling reagents. The functionalized nanoparticles were coated with fucoidan, a natural polysaccharide, using two strategies that yielded 12.4% and 33.9% fucoidan content. The resulting nanoparticles showed an adequate porosity (similar to 300 m(2)/g) to enable their use as carrier for a cytostatic agent, irinotecan. Release studies demonstrated pH-dependent kinetics with complete drug release in phosphate buffer solution pH 5.5, which mimicked the tumor environment. Cytotoxicity evaluation on Caco-2 colon adenocarcinoma cells revealed enhanced anticancer efficacy resulting from synergistic effects between irinotecan and fucoidan. The fucoidan coating of MCM-Gly maintained the biocompatibility of the silica carrier in HS27 healthy cells, while confirming the preferential uptake of the coated nanoplatforms by cancer cells in hyperspectral images. These biocompatible nanoplatforms offer a promising strategy for cancer therapy with reduced systemic toxicity.

In this paper, we present an experimental technique to measure trapping forces on individual eukaryotic cells by combining optical tweezers and dielectrophoresis. A custom 3D-printed microfluidic chip was developed to integrate these forces, allowing for direct measurement. The tweezers trapping force was determined by applying a ramped dielectrophoretic force on a trapped cell and recording the voltage at which the cell escapes. This cost-effective, robust method circumvents limitations of traditional force measurement techniques, and it senses intrinsic optical properties variations between cell types. We validated this method on two different cell lines (NIH3T3 and Caco-2) showing its applicability on large inhomogeneous cells. The effectiveness of our system in measuring forces on single cells could offer new insights into their biomechanical properties and interactions.

An integrated platform for single-cell biophysical characterization is presented. Combining dielectrophoresis (DEP) and optical tweezers (OT) within a single experiment, this approach enables the extraction of both electric properties and optical trap stiffness from individual, living cells in suspension without the need for external calibration beads, labels, or adherence to surfaces. Unlike traditional population-based DEP methods, which average over large cell ensembles and obscure cellular heterogeneity, the presented single-cell approach provides precise DEP spectra and allows direct computation of electric parameters such as membrane conductivity, permittivity, and cytoplasmic conductivity. The method is compatible with structurally and optically complex particles, such as living cells, overcoming the limitations of calibration procedures designed for spherical, homogeneous particles. It supports repeated testing of the same cell, facilitating dynamic studies of cellular responses to chemical or physical perturbations. OT stiffness measurements are performed directly on nonadherent cells that otherwise would be excluded from surface-based assays. The system includes an open-source software for data acquisition, automated image-based analysis, and OT and DEP forces computing. It is compatible with various electrode geometries, making it broadly adaptable to different experimental designs. Overall, this platform offers a robust, label-free method for high-resolution, single-cell electric and optic profiling, expanding the capabilities of DEP and OT in fundamental research, diagnostics, and bioengineering applications.

The synergistic piezoelectric and osteoconductive properties of barium titanate (BT) and hydroxyapatite (HA) could stir the development of a new generation of synthetic bone graft substitutes, with capability for rapid and safe osseointegration. The research focused on two concurrent approaches for coupling the BT and HA materials: (i) conventional sintering of BT-HA powder mixtures; and (ii) functionalization of pre-sintered BT with HA coatings using magnetron sputtering (MS). Irrespective of the BT/HA ratios ranging from 95/5 to 80/20 wt.%, nanocrystalline or highly-crystallized nature of the powders, sub-micron- or micron-sized particle dimensions, and sintering temperature, it was observed that the BT-HA reactivity cannot be prevented above 800 °C. At higher temperatures in the range of 1000 – 1300 °C, HA undergoes decomposition and extensively reacts with BT, leading to the formation of several secondary phases such as CaTiO3, Ba2Ca(PO4)2, BaCa6(PO4)4O, BaCa(PO3)4, and β-Ca2P2O7. As a consequence, the cytocompatibility assessed in fibroblast and osteoblast cell cultures, as well as the piezoelectric response, were significantly altered.Applying HA coatings by MS to the sintered BT ceramics successfully preserved their piezoelectric properties, while also providing an unaltered cytocompatible and osteogenic-prone surface. The HA coatings were fully crystallized at post-deposition annealing temperatures of 550 and 700 °C, achieving crystalline qualities comparable to HA powders sintered at 1100 and 1200 °C, respectively. No reactivity events between BT and HA were observed. Partial reactivity was only noticeable upon annealing at 1000 °C. Therefore, it is suggested that the HA coating of BT is effective in seamlessly coupling the piezoelectric and osteogenic properties of the two constituents without compromise.

Age-related macular degeneration (AMD) is a prevalent ocular pathology affecting mostly the elderly population. AMD is characterized by a progressive retinal pigment epithelial (RPE) cell degeneration, mainly caused by an impaired antioxidative defense. One of the AMD therapeutic procedures involves injecting healthy RPE cells into the subretinal space, necessitating pure, healthy RPE cell suspensions. This study aims to electrically characterize RPE cells to demonstrate a possibility using simulations to separate healthy RPE cells from a mixture of healthy/oxidized cells by dielectrophoresis. BPEI-1 rat RPE cells were exposed to hydrogen peroxide to create an in-vitro AMD cellular model. Cell viability was evaluated using various methods, including microscopic imaging, impedance-based real-time cell analysis, and the MTS assay. Healthy and oxidized cells were characterized by recording their dielectrophoretic spectra, and electric cell parameters (crossover frequency, membrane conductivity and permittivity, and cytoplasm conductivity) were computed. A COMSOL simulation was performed on a theoretical microfluidic-based dielectrophoretic separation chip using these parameters. Increasing the hydrogen peroxide concentration shifted the first crossover frequency toward lower values, and the cell membrane permittivity progressively increased. These changes were attributed to progressive membrane peroxidation, as they were diminished when measured on cells treated with the antioxidant N-acetylcysteine. The changes in the crossover frequency were sufficient for the efficient separation of healthy cells, as demonstrated by simulations. The study demonstrates that dielectrophoresis can be used to separate healthy RPE cells from oxidized ones based on their electrical properties. This method could be a viable approach for obtaining pure, healthy RPE cell suspensions for AMD therapeutic procedures.

The therapeutic use of mesenchymal stem cells (MSCs) becomes more and more important due to their potential for cell replacement procedures as well as due to their immunomodulatory properties. However, protocols for MSCs differentiation can be lengthy and may result in incomplete or asynchronous differentiation. To ensure homogeneous populations for therapeutic purposes, it is crucial to develop protocols for separation of the different cell types after differentiation. In this article we show that, when MSCs start to differentiate towards adipogenic or osteogenic progenies, their dielectrophoretic behavior changes. The values of cell electric parameters which can be obtained by dielectrophoretic measurements (membrane permittivity, conductivity, and cytoplasm conductivity) change before the morphological features of differentiation become microscopically visible. We further demonstrate, by simulation, that these electric modifications make possible to separate cells in their early stages of differentiation by using the dielectrophoretic separation technique. A label free method which allows obtaining cultures of homogenously differentiated cells is thus offered.

Backgound: Type 2 diabetes mellitus (T2DM) is a major cardiovascular risk factor. Nitric oxide (NO) is one of the many molecules that regulate vascular tone, and red blood cells (RBCs) are known to play an important role in adjusting cardiac function through NO export from RBCs. Our study prospectively investigated the L-arginine (L-arg)–nitric oxide (NO) metabolic pathway in the erythrocytes and plasma of subjects with T2DM. Methods: RBCs and plasma were collected from patients with T2DM (n = 10), at first clinical onset (baseline) and after five years of disease evolution (follow-up). L-arg content was assayed by competitive enzyme-linked immunoassay. Arginase activity and nitrate/nitrite levels were measured using spectrophotometry. Results: When compared to baseline, L-arg content decreased in RBCs and remained similar in the plasma; NO production decreased in RBCs and the plasma; and arginase activity was lower in RBCs and increased in plasma. Conclusions: The L-arg/NO metabolic pathway decreases in the RBCs of patients with T2DM five years after the first clinical onset. The persistent decrease in RBCs’ arginase activity fails to compensate for the sustained decrease in RBCs’ NO production in the diabetic environment. This pilot study indicates that the NO-RBC pool is depleted during the progression of the disease in the same cohort of T2DM patients.

Dielectrophoretic (DEP) cell separation, which utilizes electric fields to selectively manipulate and separate cells based on their electrical properties, has emerged as a cutting-edge label-free technique. DEP separation techniques rely on differences in the electrical and morphological properties of cells, which can be obtained by a thorough analysis of DEP spectra. This article presents a novel platform, named OpenDEP, for acquiring and processing DEP spectra of suspended cells. The platform consists of lab-on-a-chip and open-source software that enables the determination of DEP spectra and electric parameters. The performance of OpenDEP was validated by comparing the results obtained using this platform with the results obtained using a commercially available device, 3DEP from DEPtech. The lab-on-a-chip design features two indium tin oxide-coated slides with a specific geometry, forming a chamber where cells are exposed to an inhomogeneous alternating electric field with different frequencies, and microscopic images of cell distributions are acquired. A custom-built software written in the Python programing language was developed to convert the acquired images into DEP spectra, allowing for the estimation of membrane and cytoplasm conductivities and permittivities. The platform was validated using two cell lines, DC3F and NIH 3T3. The OpenDEP platform offers several advantages, including easy manufacturing, statistically robust computations due to large cell population analysis, and a closed environment for sterile work. Furthermore, continuous observation using any microscope allows for integration with other techniques.

Mouse fibroblasts were exposed to buffers containing various concentrations of DMSO and dielectrophoretic spectra were acquired. Based on these spectra, electric parameters of the cell (cytosol conductivity, membrane conductivity and membrane permittivity) were computed. A decrease of all electrical parameters was observed with the increase of the DMSO concentration. Results were analyzed in terms of DMSO perturbation of the structured water layers adjacent to the membrane and of lipid packing.

Chronic nandrolone decanoate exposure is associated with a modified redox status in the favor of oxidants. Paraoxonase and myeloperoxidase activities are considered sensitive biomarkers of oxidative stress and inflammation, being also predictors for coronary artery disease. The aim of our study was to investigate the impact of chronic nandrolone administration on paraoxonase and myeloperoxidase activities in Wistar rats. 16 Wistar rats were divided into two groups (n=8 for each of them): nandrolone decanoate (A) treated group and control group (C). After 12 weeks of nandrolone decanoate administration we analyzed several plasma oxidative stress parameters: TROLOX Equivalent Antioxidant Activity, paraoxonase and myeloperoxidase activities, total thiols, and advanced oxidation protein products. Our results showed a significant decreased total antioxidant activity in nandrolone treated group compared with controls, while paraoxonase activity and total thiols levels were significantly increased. Myeloperoxidase activity and advanced oxidation protein products were not significantly different between the two studied groups. In conclusion decreased plasma total antioxidant activity suggested increased oxidative stress induced by chronic high doses of nandrolone. Despite oxidative stress presence high doses of nandrolone induced also increased plasma paraoxonase activity.

Electropermeabilization of the cell membrane is a technique used to facilitate penetration of impermeant molecules into cells. Although there are studies regarding the mechanism of processes occurring after electropermeabilization, the relationship between electropermeabilization and associated phenomena (e.g. generation of reactive oxygen species, endocytosis, lipid peroxidation, etc.) is yet to be elucidated. This work aimed to get information on the changes in the packing of the bilayer lipids and their peroxidation induced by application of electroporation pulses. We used a specially designed system of electrodes which allowed performing electropermeabilization of cells in suspension simultaneously with time-dependent measurements of fluorescence and temperature. The kinetics of membrane packing and production of reactive oxygen species were studied using various conductivity buffers (0.01, 0.04 and 0.14 S/m) and different number of 1 kV/cm bipolar pulses (1-50). Two categories of effects were observed: a thermal effect, consisting in an increased bilayer disorder (a deeper penetration of water into the hydrophobic core), and a nonthermal effect, leading to a higher degree of lipids packing, the latter being attributed to a peroxidation process. An analysis of the permeabilization conditions in which one of these two processes predominates was performed. (c) 2020 Elsevier B.V. All rights reserved.

The present study concerns the in vitro oxidative stress responses of non-malignant murine cells exposed to surfactant-tailored ZnO nanoparticles (NPs) with distinct morphologies and different levels of manganese doping. Two series of Mn-doped ZnO NPs were obtained by coprecipitation synthesis method, in the presence of either polyvinylpyrrolidone (PVP) or sodium hexametaphosphate (SHMTP). The samples were investigated by powder X-ray Diffraction, Transmission Electron Microscopy, Fourier-Transform Infrared and Electron Paramagnetic Resonance spectroscopic methods, and N 2 adsorption–desorption analysis. The observed surfactant-dependent effects concerned: i) particle size and morphology; ii) Mn-doping level; iii) specific surface area and porosity. The relationship between the surfactant dependent characteristics of the Mn-doped ZnO NPs and their in vitro toxicity was assessed by studying the cell viability, intracellular reactive oxygen species (ROS) generation, and DNA fragmentation in NIH3T3 fibroblast cells. The results indicated a positive correlation between the specific surface area and the magnitude of the induced toxicological effects and suggested that Mn-doping exerted a protective effect on cells by diminishing the pro-oxidative action associated with the increase in the specific BET area. The obtained results support the possibility to modulate the in vitro toxicity of ZnO nanomaterials by surfactant-controlled Mn-doping.

The electroporation of cells is nowadays used for a large variety of purposes, from basic research to cancer therapy and food processing. Understanding molecular mechanisms of the main processes involved in electroporation is thus of significant interest. In the present work, we propose an experimental system to record in real time the evolution of any cell parameter which can be evaluated by fluorescence (before, during and after application of the electroporation pulses to cells in suspension). The system is based on the design of adequate electroporation electrodes, compatible with a standard spectrofluorometer cuvette housing. The electric field intensity generated when pulses are applied was carefully characterized for different geometries of the electrodes, to choose a construction ensuring the greatest homogeneity of the field in combination with the best possible illumination of the sample. As an example of the method’s application, we present here generalized polarization kinetics for a varying number of electroporation pulses applied to a cell suspension; the general polarization parameter is strongly correlated to water presence in the hydrophobic membrane core. The system may be used for many other fluorescence measurements useful for the characterization of the electroporation process.

Common and durum wheat are widelly used for many purposes and the prices are significantly different for each wheat type, thus efficient methods for detection of accidental or intentional admixtures of common wheat to durum wheat products, and vice-versa, are required. The aim of this study is to identify some characteristics of the Triticum aestivum genetic material witch will be useful in developing methods for determination of the quality and authenticity of this species. Melting, DNA methylation and RAPD analysis can be used to discriminate between wheat species/varieties.

Supraphysiological administration of anabolic androgenic steroids has been linked to increased blood pressure. The widely distributed amino acid taurine seems to be an effective depressor agent in drug-induced hypertension. The purpose of this study was to assess the impact of chronic high dose administration of nandrolone decanoate (DECA) and taurine on blood pressure in rats and to verify the potentially involved mechanisms. The study was conducted in 4 groups of 8 adult male Wistar rats, aged 14 weeks, treated for 12 weeks with: DECA (A group); vehicle (C group); taurine (T group), or with both drugs (AT group). Systolic blood pressure (SBP) was measured at the beginning of the study (SBP1), 2 (SBP2) and 3 months (SBP3) later. Plasma angiotensin-converting enzyme (ACE) activity and plasma end products of nitric oxide metabolism (NOx) were also determined. SBP3 and SBP2 were significantly increased compared to SBP1 only in the A group (P<0.002 for both). SBP2, SBP3 and ACE activity showed a statistically significant increase in the A vs C (P<0.005), andvs AT groups (P<0.05), while NOx was significantly decreased in the A and AT groups vs controls (P=0.01). ACE activity was strongly correlated with SBP3 in the A group (r=0.71, P=0.04). These findings suggest that oral supplementation of taurine may prevent the increase in SBP induced by DECA, an effect potentially mediated by angiotensin-converting enzyme.

Recent studies have unveiled the unique roles of extracellular vesicles (EVs) in various cellular processes including protein degradation, transport, and intercellular communication. However, the EVs of Chinese hamster ovary (CHO) cells, the workhorse of biologics manufacturing, have not been well-characterized despite their significant roles in protein production. Herein, we successfully isolated CHO EVs from CHO fed-batch cultures and identified their messenger RNA (mRNA) and micro RNA (miRNA) contents through next-generation sequencing. We found that mRNAs corresponding to oxidative phosphorylation were highly enriched in microvesicles (large EVs) but absent in exosomes (small EVs). We also found that both large EVs and small EVs had enriched mRNA species corresponding to key signaling pathways for cell proliferation, survival, and growth, including the TGFβ and PI3K/Akt pathways. In addition, the enrichment of miR-196a-5p in both small EVs and large EVs suggests an anti-apoptotic and proliferative function for EVs through intercellular communication. The identification of these mRNAs and miRNAs associated with cell growth and survival sheds light on the potential role of extracellular vesicles in the context of biologics manufacturing and may help further optimize CHO biologics production.

Consumers are increasingly concerned about the health aspects of food preparation. There is data to support that foods treated with microwaves (MW) develop harmful substances. Vegetable oils are susceptible to lipid peroxidation due to the chemical structure, but also contain antioxidant compounds. Peroxidation products can have harmful effects for human health. The aim of the study was to evaluate the variation of total antioxidant capacity and antioxidant degradation in relation to lipid peroxides formation during conventional and microwave oils heating. Sunflower, corn, soybean, palm and a mixed oil (sunflower, grape, flaxseed and rice oil) were purchased from the local market. To simulate home cooking, different exposure times were tested 5, 10 and 15 min. For all samples, and for each exposure time, Trolox Equivalent Antioxidant Capacity (TEAC), lipid peroxides as thiobarbituric reactive substances (TBARS), conjugated dienes, vitamin E as α-tocopherol, flavonoids and total polyphenols were determined. After 15 minutes heating the best retention for vitamin E was observed for the palm oil (MW 12 mM/l), and corn oil (convection 17 mM/l). The highest total antioxidant capacity after heating belonged to mixed & palm oil (MW 4.6 eq. TROLOX/l) and soybean oil (convection 7.5 mM eq. TROLOX/l). Highest levels of TBARS belonged to the mixed oil (convection 54 uM/l) and soybean oils (MW 83 uM/l). Highest conjugated dienes levels were observed for soybean oil (MW 17 mM/l) and sunflower oil (convection 22 mM/l). The best retention for polyphenols was found in the mixed and sunflower oils (MW and convection 0.80.9 mg/ml. The retention of flavonoid was very small in all oils and the highest amount was found in corn and soybean oil (0.04 mg/ml). During the heating of edible vegetable oils commonly used, total antioxidant capacity along with the amount of vitamin E, flavonoids and polyphenols decreases, while the amount of lipid peroxidation products increases and these processes are more intense when using microwave heating than when using convection heating.

Individual phenolic compounds, total phenolic content and antioxidant potential were assessed in kernels, oils and bagasse pellets (residues of oil pressing) of different walnut cultivars. Twenty-seven phenolic compounds were detected in kernels and pellets conducting high-performance liquid chromatography–tandem mass spectrometry. The main polyphenolic subclass comprised hydrolysable tannins, which accounted approximately 60.80% (kernels) and 61.66% (pellets) of the total phenolics identified (TPI). Walnut oil was poor in phenolics and contained only six different compounds but due to their low content (from 0.15 to 1.44 μg g− 1) just two compounds have been identified. Glansreginin A and glansreginin B were detected in all analyzed walnut products. A comparison of average amount of total phenolic content revealed that walnut oil contains as much as 154 fold less phenolics (0.05 mg GAE g− 1 FW) compared to kernels (7.7 mg GAE g− 1 FW) or pellets (7.9 mg GAE g− 1 FW).

Estrogens role in schizophrenia patients is a subject, which has gained an increased attention from the medical community. Estrogens have been shown to inhibit dopamine actions, improve neuronal regeneration, and overall, have a protective role in the pathology of schizophrenia. The adjunctive estrogen therapy for men is currently under debate. Antipsychotic medication is known to influence the hypothalamo-hypophyseal - gonadal axis by inducing variable degrees of hyperprolactinemia. Several studies have found that some of the atypical antipsychotics lower cortisol levels in patients and also in healthy controls. We have investigated the effects of clozapine and risperidone on estradiol levels in men with schizophrenia. We have also evaluated the levels of prolactin and cortisol, taking into account the possible influence of antipsychotic drugs on both these hormones. Both prolactin and cortisol also have the potential to regulate sexual hormones biosynthesis. Our study found decreased estradiol levels in men with schizophrenia treated with clozapine and risperidone, while prolactin levels were increased only in the risperidone treated group. Cortisol levels are not statistically significant different between groups.

Objective In the current study, we investigated the impact of oxidant overload on  plasma total antioxidant capacity (TEAC) and antioxidant gap (GAP) in seven groups of children with different types of mitochondrial dysfunction and 20 control subjects. Methods 101 children with suspicion of mitochondrial disease presented at Psichiatry Hospital Dr. Alexandru Obregia and 20 children  presented at Alfred Rusescu childrenu0027s hospital have been enrolled in the study. Eight groups have been considered: group 1- with global development disorder (n=25), group 2 – with epileptic encephalopathy (n=22), group 3- with myoclonic epilepsy (n=9), group 4 - with cerebellar syndrome (n=10), and group 5 – with extrapyramidal syndrome (n=11),  group 6 – with demyelinating disease (n=13), group 7– with cranial nerves defects (n=11) and group 8– controls (n=20). Concentrations of albumin, uric acid, TEAC and GAP have been measured in  plasma  samples. Conclusion Significaly increased TEAC and GAP levels (pu003c0.001) were observed in controls compared with all groups of children with suspicion of mitochondrial disease. No significant changes were observed in TEAC and GAP levels between the  groups of children with suspicion of mitochondrial disease. Our results are sustaining increased oxidative stress in children with suspicion of mitochondrial disease.