Novel Benzo[a]phenoxazinium Chlorides Functionalized with Sulfonamide Groups as NIR Fluorescent Probes for Vacuole, Endoplasmic Reticulum, and Plasma Membrane Staining.

The demand for new fluorophores for different biological target imaging is increasing. Benzo[a]phenoxazine derivatives are fluorochromophores that show promising optical properties for bioimaging, namely fluorescent emission at the NIR of the visible region, where biological samples have minimal fluorescence emission. In this study, six new benzo[a]phenoxazinium chlorides possessing sulfonamide groups at 5-amino-positions were synthesized and their optical and biological properties were tested. Compared with previous probes evaluated using fluorescence microscopy, using different S. cerevisiae strains, these probes, with sulfonamide groups, stained the vacuole membrane and/or the perinuclear membrane of the endoplasmic reticulum with great specificity, with some fluorochromophores capable of even staining the plasma membrane. Thus, the addition of a sulfonamide group to the benzo[a]phenoxazinium core increases their specificity and attributes for the fluorescent labeling of cell applications and fractions, highlighting them as quite valid alternatives to commercially available dyes.

IL-15Rα membrane anchorage in either cis or trans is required for stabilization of IL-15 and optimal signaling.

Interleukin (IL)-15 plays an important role in the communication between immune cells. It delivers its signal through different modes involving three receptor chains: IL-15Rα, IL-2Rß and IL-2Rγc. The combination of the different chains result in the formation of IL-15Rα/IL-2Rß/γc trimeric or IL-2Rß/γc dimeric receptors. In this study, we have investigated the role of the IL-15Rα chain in stabilizing the cytokine in the IL-2Rß/γc dimeric receptor. By analyzing the key amino acid residues of IL-15 facing IL-2Rß, we provide evidence of differential interfaces in the presence or in the absence of membrane-anchored IL-15Rα. Moreover, we found that the anchorage of IL-15Rα to the cell surface regardless its mode of presentation - i.e. cis or trans - is crucial for complete signaling. These observations show how the cells can finely modulate the intensity of cytokine signaling through the quality and the level of expression of the receptor chains.

Mechanistic and Structural Insights on the IL-15 System through Molecular Dynamics Simulations.

Interleukin 15 (IL-15), a four-helix bundle cytokine, is involved in a plethora of different cellular functions and, particularly, plays a key role in the development and activation of immune responses. IL-15 forms receptor complexes by binding with IL-2Rß- and common γ(γc)-signaling subunits, which are shared with other members of the cytokines family (IL-2 for IL-2Rß- and all other γc- cytokines for γc). The specificity of IL-15 is brought by the non-signaling α-subunit, IL-15Rα. Here we present the results of molecular dynamics simulations carried out on four relevant forms of IL-15: its monomer, IL-15 interacting individually with IL-15Rα (IL-15/IL-15Rα), with IL-2Rß/γc subunits (IL-15/IL-2Rß/γc) or with its three receptors simultaneously (IL-15/IL-15Rα/IL-2Rß/γc). Through the analyses of the various trajectories, new insights on the structural features of the interfaces are highlighted, according to the considered form. The comparison of the results with the experimental data, available from X-ray crystallography, allows, in particular, the rationalization of the importance of IL-15 key residues (e.g. Asp8, Lys10, Glu64). Furthermore, the pivotal role of water molecules in the stabilization of the various protein-protein interfaces and their H-bonds networks are underlined for each of the considered complexes.

Insights into the reaction mechanism of 3-O-sulfotransferase through QM/MM calculations.

3-O-Sulfotransferase (3-OST) is one of the enzymes involved in heparan sulfate (HS) biosynthesis. HSs are polysaccharides with variable patterns of sulfation and acetylation that serve as entry receptors for herpes simplex virus type 1 (HSV-1). 3-OST is responsible for the transfer of a sulfate group from 3'-phosphoadenosine-5'-phosphosulfate (PAPS) to glucosamine units of HS. In this work, the catalytic mechanism of 3-OST was studied with atomic detail, using computational methods. We investigated the protonation state of key residues using the H++ web-based pKa prediction tool and molecular dynamics (MD) simulations and estimated the most relevant protonation state of the catalytic residues during catalysis. Catalytic histidine (His186) is predominantly protonated, while catalytic aspartate and glutamate (Asp189 and Glu184) are predominantly deprotonated. Subsequently, to study the catalytic mechanism, we applied a QM/MM method at the ONIOM(B3LYP/6-31G(d):ff94) level, starting from three geometries extracted from the 3, 6 and 8 ns point on the MD simulation. The results show that the reaction mechanism of 3-OST occurs by a single elementary step, consisting of an associative SN2 transfer of the sulfate group from PAPS to the HS glucosamine units, with the transfer of a proton from glucosamine to the catalytic Glu184. The activation free energies for this reaction were determined at the ONIOM(M06-2X-D3/6-311++G(2d,2p):ff94//B3LYP/6-31G(d):ff94) level of theory. Despite the free energy differences among the three conformations (10.2, 20.9 and 16.1 kcal mol(-1)), our results are consistent with the upper limit determined experimentally for the full cycle (20.4 kcal mol(-1)). The data obtained in this study will be useful for further studies on the inhibition of this enzyme, which is a useful target for drugs that block HSV-1 viral infections.

The Potential of Electrospun Membranes in the Treatment of Textile Wastewater: A Review.

Water security and industrial wastewater treatment are significant global concerns. One of the main issues with environmental contamination has been the discharge of dye wastewater from the textile and dye industries, contributing to an ever-growing problem with water pollution, poisoning water supplies, and harming the ecosystem. The traditional approach to wastewater treatment has been found to be inefficient, and biosorption techniques and mechanisms have been proven to be a successful replacement for conventional methods. Recent developments have led to the recognition of fibrous materials as an environmentally friendly option with broad application in several industries, including wastewater treatment. This review explores the potential of fibrous materials produced by the electrospinning technique as adsorbents for wastewater treatment, while at the same time, for the removal of adsorbates such as oil, dyes, heavy metals, and other substances, as reported in the literature. Textile wastewater filtering structures, produced by electrospinning, are summarized and the use of synthetic and natural polymers for this purpose is discussed. The limitations of electrospun textile wastewater filtering structures are also mentioned. Electrospun nanofibrous membranes appear to be a very promising route to filter textile wastewater and therefore contribute to water reuse and to reducing the contamination of water courses.

Amino-Alcohol Organic-Inorganic Hybrid Sol-Gel Materials Based on an Epoxy Bicyclic Silane: Synthesis and Characterization.

Organic-inorganic hybrids (OIHs) are a type of material that can be obtained using the sol-gel process and has the advantages of organic and inorganic moieties in a single material. Polyetheramines have been widely used in the preparation of this type of material, particularly in combination with epoxy-based alkoxysilanes. Nevertheless, epoxyciclohexylethyltrimethoxysilane (ECHETMS) is a promising alkoxysilane with an epoxy terminal group that is quite unexplored. In this work, four novel OIH materials were synthesized using the sol-gel method. The OIHs were based on Jeffamines® of different molecular weights (D-230, D-400, ED-600, and ED-900), together with ECHETMS. The materials were characterized using multinuclear solid state NMR, FTIR, BET, UV/Vis spectroscopy, EIS, and TGA. The influence of the Jeffamine molecular weight and the suitability of these materials to act as a supporting matrix for heteroaromatic probes were assessed and discussed. The materials show interesting properties in order to be applied in a wide range of sensing applications.

Hybrid Sol-Gel Matrices Doped with Colorimetric/Fluorimetric Imidazole Derivatives.

Organic-inorganic hybrids (OIH) are materials that can be easily synthesized by the sol-gel method and combine the advantages of organic and inorganic moieties within a single polymeric matrix. Imidazole derivatives are versatile organic compounds that can change their optical properties with the variation of pH due to the protonation or deprotonation of the nitrogen atoms. This work reports the preparation of different OIHs doped with different contents of two imidazole compounds (3a,b). The obtained materials were characterized structurally by FTIR, and the dielectric properties were studied by electrochemical impedance spectroscopy. The optical properties were studied by UV-Vis absorption and fluorescence spectroscopies. The FTIR analysis showed that the presence of the imidazole does not change the structural properties of the matrices. The normalized resistance values obtained for the doped matrices ranged between 8.57 and 9.32 Ω cm2, all being higher than the undoped matrix. The σ ranged between 9.49 and 10.28 S cm-1, being all higher than the pure OIH samples. Compound 3a showed a maximum absorption peak at 390 nm, which is present in the OIH spectra, proving the presence of the compound. In the case of compound 3b, a maximum absorption wavelength at 412 nm was found, and the compound peak was not clear, which may indicate that an interaction between the compound and the matrix occurred. A synergetic effect between the intrinsic emission of the matrix and the fluorescence of 3a is found on the OIH-doped matrices.

Organic-inorganic hybrid sol-gel materials doped with a fluorescent triarylimidazole derivative.

The development of sensors for pH monitoring is of extreme importance in the monitoring of concrete and reinforced concrete structures. Imidazole derivatives are promising probes for pH sensing due to the amphoteric nature of their heterocyclic ring, which can be protonated/deprotonated upon pH changes. In this work, a triarylimidazole was synthesised and used as a dopant in an organic-inorganic hybrid (OIH) sol-gel matrix to obtain a pH-sensitive membrane for further application in optical fibre sensors (OFS). The triarylimidazole probe shows fluorimetric response in pH between 9 and 13, which is the desired range for monitoring carbonation of concrete. This degradation process lowers the highly alkaline pH of concrete (12.5-13) to values below 9, which creates favourable conditions for corrosion of concrete reinforcement. The OIH membranes used were based on Jeffamine THF170 and 3-glycidoxypropytrimethoxysilane precursors, which had already been shown to be suitable and resistant in contact with cement-based materials. The OIHs were doped with three different contents of the triarylimidazole and the structural, dielectric, thermal and optical properties of the pure and doped OIH materials were evaluated. The structural analysis showed that the presence of the triarylimidazole did not change the structural properties of the OIH material. Electrochemical impedance spectroscopy showed that in the doped samples the conductivity increased with the imidazole concentration. The ε r obtained for the doped samples ranged approximately from 11 to 19 and for the pure matrices was 8. Thermal analysis showed that these materials are stable up to 350 °C and that the presence of the probe did not change that feature. The optical properties showed that the prepared OIH materials have promising properties to be used as pH sensitive fluorimetric probes.

Optical Fiber Sensors for Biocide Monitoring: Examples, Transduction Materials, and Prospects.

Antifouling biocides are toxic to the marine environment impacting negatively on the aquatic ecosystems. These biocides, namely, tributyltin (TBT) and Cu(I) compounds, are used to avoid biofouling; however, their toxicity turns TBT and Cu(I) monitoring an important health issue. Current monitoring methods are expensive and time-consuming. This review provides an overview of the actual state of the art of antifouling paints' biocides, including their impact and toxicity, as well as the reported methods for TBT and Cu(I) detection over the past decade. The principles of optical fiber sensors (OFS) applications, with focus on environmental applications, and the use of organic chemosensors in this type of sensors are debated. The multiplexing ability of OFS and their application on aquatic environments are also discussed.

PDMS Based Hybrid Sol-Gel Materials for Sensing Applications in Alkaline Environments: Synthesis and Characterization.

Nowadays, concrete degradation is a major problem in the civil engineering field. Concrete carbonation, one of the main sources of structures' degradation, causes concrete's pH to decrease; hence, enabling the necessary conditions for corrosion reinforcement. An accurate, non-destructive sensor able to monitor the pH decrease resistant to concrete conditions is envisaged by many researchers. Optical fibre sensors (OFS) are generally used for concrete applications due to their high sensitivity and resistance to external interferences. Organic-inorganic hybrid (OIH) films, for potential functionalization of OFS to be applied in concrete structures, were developed. Polydimethylsiloxane (PDMS) based sol-gel materials were synthesized by the formation of an amino alcohol precursor followed by hydrolysis and condensation. Different ratios between PDMS and (3-aminopropyl)triethoxysilane (3-APTES) were studied. The synthesized OIH films were characterized by Fourier-transformed infrared spectroscopy (FTIR), UV-Vis spectroscopy, electrochemical impedance spectroscopy (EIS) and thermogravimetric analysis (TGA). The OIH films were doped with phenolphthalein (Phph), a pH indicator, and were characterized by UV-Vis and EIS. FTIR characterization showed that the reaction between both precursors, the hydrolysis and the condensation reactions occurred successfully. UV-Vis characterization confirmed the presence of Phph embedded in the OIH matrices. Dielectric and thermal properties of the materials showed promising properties for application in contact with a high alkaline environment.

Metabolic remodeling associated with subchronic doxorubicin cardiomyopathy.

Doxorubicin (Adriamycin) is a potent and broad-spectrum antineoplastic agent, the clinical utility of which is restricted by a cumulative and progressive cardiomyopathy that develops with repeated dosing. Fundamental to the cardiac failure is an interference with mitochondrial respiration and inhibition of oxidative phosphorylation. Global gene expression arrays in cardiac tissue indicate that inhibition of mitochondrial oxidative phosphorylation by doxorubicin (DOX) is accompanied by a decreased expression of genes related to aerobic fatty acid oxidation and a corresponding increase in expression of genes involved in anaerobic glycolysis, possibly as an alternate source for ATP production. The aim of this investigation was to determine whether this is also manifest at the metabonomic level as a switch in metabolic flux in cardiac tissue, and whether this can be averted by co-administering the cardioprotective drug, dexrazoxane (DZR). (13)C-isotopomer analysis of isolated perfused hearts from male Sprague-Dawley rats receiving 6 weekly s.c. injections of 2mg/kg DOX demonstrated a shift from the preferential oxidation of fatty acids to enhanced oxidation of glucose and lactate plus pyruvate, indicative of a compensatory shift towards increased pyruvate dehydrogenase activity. Substrate-selective isotopomer analysis combined with western blots indicate an inhibition of long-chain fatty acid oxidation and not MCAD activity or fatty acyl-carnitine transport. Co-administering DZR averted many treatment-related changes in cardiac substrate metabolism, consistent with DZR being an effective cardioprotective agent against DOX-induced cardiomyopathy. This switch in substrate metabolism resembles that described for other models of cardiac failure; accordingly, this change in metabolic flux may represent a general compensatory response of cardiac tissue to imbalances in bioenergetic demand and supply, and not a characteristic unique to DOX-induced cardiac failure itself.