A study on the coordinative versatility of new N,S-donor macrocyclic ligands: XAFS, and Cu2+ complexation thermodynamics in solution

We investigated, both in the solid state and in aqueous solution, the coordination environment and stability behavior of four macrocyclic ligands (three N(2)S(2) and one N(3)S(2)) and of the corresponding Cu(II) complexes. The structural characterization in the solid state of the copper derivatives was performed by X-Ray Absorption Spectroscopy. Copper is found to be 4-fold coordinated with a CuN(2)S(2) environment with different Cu-S distances depending on the size of the macrocyclic ring. The EXAFS technique has indicated that nitrogen and sulfur atoms are more preferable to oxygen atoms as donor systems, without the evidence of coordination of the carboxylic moieties to copper in the first shell. The joint EXAFS and XANES study of the copper(II) complex with the N(3)S(2) ligand confirms the 4-fold coordination with an additional, long Cu-N interaction. The Cu(2+) complexation constants for one ligand were determined in aqueous solution. The results indicate that the species [CuL], although isolated in the solid state, is not the most abundant at the pH of blood serum. Instead, at pH 7.4 the protonated [Cu(HL)](+) species was found to be the most relevant. The behaviour of the copper complexes in the presence of the strong copper chelating bioagent human serum albumin was also examined in order to gain information on the stability of these compounds in biological fluids.     

Macroscopic dynamics as reporter of mesoscopic organization: the Belousov-Zhabotinsky reaction in aqueous layers of DPPC lamellar phases

The propagation of traveling chemical waves in the excitable Belousov-Zhabotinsky (BZ) system when performed in the presence of 1,2-dipalmitoyl-sn-glycero-3-phosphatidyl choline (DPPC) bilayers responds sensitively to the phospholipid content. The characteristic features of wave propagation, such as spiral pitch, rotation period, and size of the spiral core region, show two regions of different behavior, one below and the other above a DPPC content of 12.5% (w/w) thus suggesting a transition in the organization of the lipid domains at a DPPC content of ～12.5% (w/w). This transition is supported by small-angle X-ray scattering data, which show pronounced changes in the coherence lengths of the lyotropic smectic domains. Thus, the dynamics of the chemical system occurring at a macroscopic length scale reflects the organization of the water/lipid domains which extend over mesoscopic lengths. These findings indicate that in the BZ/DPPC system, there is an interaction between processes that occurs at length scales differing by as much as 3 orders of magnitude.     

An integrated approach for the systematic evaluation of polymeric nanoparticles in healthy and diseased organisms

Innovative strategies that utilize nanoparticles (NPs) for a better delivery of drugs and to improve their therapeutic efficacy have been widely studied in many clinical fields, including oncology. To develop safe and reliable devices able to reach their therapeutic target, a hierarchical characterization of NP interactions with biological fluids, cells, and whole organisms is fundamental. Unfortunately, this aspect is often neglected and the development of standardized characterization methods would be of fundamental help to better elucidate the potentials of nanomaterials, even before the loading of the drugs. Here, we propose a multimodal in vitro/in vivo/ex vivo platform aimed at evaluating these interactions for the selection of the most promising NPs among a wide series of materials. To set the system, we used non-degradable fluorescent poly(methyl-methacrylate) NPs of different sizes (50, 100, and 200 nm) and surface charges (positive and negative). First we studied NP stability in biological fluids. Then, we evaluated NP interaction with two cell lines of triple-negative breast cancer (TNBC), 4T1, and MDA-MB231.1833, respectively. We found that NPs internalize in TNBC cells depending on their physico-chemical properties without toxic effects. Finally, we studied NP biodistribution in terms of tissue migration and progressive clearance in breast-cancer bearing mice. The use of highly stable poly(methyl-methacrylate) NPs enabled us to track them for a long time in cells and animals. The application of this platform to other nanomaterials could provide innovative suggestions for the development of a systematic method of characterization to select the most reliable nanodrug candidates for biomedical applications.     

Saffron and Its Major Ingredients’ Effect on Colon Cancer Cells with Mismatch Repair Deficiency and Microsatellite Instability

Background: Colorectal cancer (CRC) is one of the most common cancers worldwide. One of its subtypes is associated with defective mismatch repair (dMMR) genes. Saffron has many potentially protective roles against colon malignancy. However, these roles in the context of dMMR tumors have not been explored. In this study, we aimed to investigate the effects of saffron and its constituents in CRC cell lines with dMMR. Methods: Saffron crude extracts and specific compounds (safranal and crocin) were used in the human colorectal cancer cell lines HCT116, HCT116+3 (inserted MLH1), HCT116+5 (inserted MSH3), and HCT116+3+5 (inserted MLH1 and MSH3). CDC25b, p-H2AX, TPDP1, and GAPDH were analyzed by Western blot. Proliferation and cytotoxicity were analyzed by MTT. The scratch wound assay was also performed. Results: Saffron crude extracts restricted (up to 70%) the proliferation in colon cells with deficient MMR (HCT116) compared to proficient MMR. The wound healing assay indicates that deficient MMR cells are doing better (up to 90%) than proficient MMR cells when treated with saffron. CDC25b and TDP1 downregulated (up to 20-fold) in proficient MMR cells compared to deficient MMR cells, while p.H2AX was significantly upregulated in both cell types, particularly at >10 mg/mL saffron in a concentration-dependent manner. The reduction in cellular proliferation was accompanied with upregulation of caspase 3 and 7. The major active saffron compounds, safranal and crocin reproduced most of the saffron crude extracts’ effects. Conclusions: Saffron’s anti-proliferative effect is significant in cells with deficient MMR. This novel effect may have therapeutic implications and benefits for MSI CRC patients who are generally not recommended for the 5-fluorouracil-based treatment.     

Influence of lipid composition on the thermotropic behavior and size distribution of mixed cationic liposomes

Cationic liposomes are studied mainly as nonviral nucleic acid delivery systems and to a lesser extent as carriers/adjuvants of vaccines and as low-molecular-weight drug carriers. It is well established that the performance and the biological activity of liposomes in general are strongly related to their physicochemical properties. We investigated the thermotropic behavior and the size distribution of mixed cationic liposomes formulated with different percentages of 1,2 dimyristoyl-sn-glycero-3-phosphatidylcholine and one of four cationic amphiphiles characterized by a pyrrolidinium headgroup with the aim of achieving a better understanding of how the molecular structure of the cationic amphiphile and its mole percentage affect the physicochemical properties of the liposomes. Multilamellar vesicles and large unilamellar vesicles were studied by differential scanning calorimetry and turbidity, respectively, to characterize the thermotropic behavior and lipid phase, whereas dynamic light scattering was used to determine size distribution. This study shows that subtle modifications in the cationic amphiphile's molecular structure and in liposome composition may have dramatic effects on the organization of the liposome bilayer and hence on the morphological and physicochemical features of the liposomes, thus being highly relevant to the biological features investigated previously.     

Multivalent calixarene-based C-fucosyl derivative: a new Pseudomonas aeruginosa biofilm inhibitor

The first example of multivalent conjugate in which four α-l-C-fucosyl units are clustered by means of a calix[4]arene platform was designed as a new potential Pseudomonas aeruginosa biofilm inhibitor. The anti-biofilm activity of the synthesized compound (6) against PAO1 strain was assayed and it was found to be dose-dependent. The presence of the fucose cluster improves the biofilm inhibitor efficiency as proven by the lower inhibitor activity of the analogous glycyl-calix[4]arene derivative (3) lacking in the fucose moieties.     

PANI-CSA: An Easy Method to Avoid ITO Photolithography in PLED Manufacturing

In order to optimize polymer light emitting diode (PLED) performances, devices with holes injected through an Indium Tin Oxide (ITO) / Polyaniline (PANI) electrode into the polymer are much more efficient than devices fabricated with the anode made only by ITO. We demonstrated that by using doped PANI as hole injection layer in a polymer light emitting diode the manufacturing process can become simpler. Indeed, the pattern of conductive layer can be produced without ITO photolithography by UV exposition. As hole transporter layer, Poly(N-vinylcarbazole) (PVK) was spin coated over the doped PANI layer and a layer of tris (8-hydroxy) quinoline aluminum (Alq₃) was then thermally evaporated so as to form the electron transport layer. To complete the device structure, Aluminum contacts were deposited onto the organic layers by vacuum evaporation at low pressure. The layers were characterized by X-ray small-angle diffraction, IR Raman and UV-Vis spectroscopies. Devices without PANI and with PANI as HIL were studied.