Elusive π-helical peptide foldamers spotted by chiroptical studies.

A series of oligomers containing alternate l-Ala and pGlu (pyroglutamic acid) both in the L and D form have been prepared and conformationally investigated by X-ray, NMR, UV/ECD, IR/VCD and molecular modelling. X-ray diffraction analysis was possible for the shortest oligomers LL-1 and LD-1. Molecular dynamics simulations of the oligomers demonstrated that the energy landscapes of the LL-series are broad. In contrast, the energy landscapes of the LD-series are characterized by well-defined minima corresponding to specific conformational structures. A single well-defined minimum exists in the energy landscape of the largest oligomer LD-8, corresponding to a precise conformation, characterized by i + 5 →i N-HO[double bond, length as m-dash]C hydrogen bonds, typical of a π-helix. ECD and VCD spectra were measured to identify the chiroptical profiles of the oligomers. The most striking element in the ECD spectra of the LD-series is their exceptionally strong intensity, which confirms that these polypeptides attain a high degree of helical order. VCD spectra for the LD-series are well reproduced by frequency calculations when π-helix folds are employed as input structures, suggesting that a symmetrical VCD couplet around 1720 cm-1 can be taken as the VCD signature of π-helices.

Tuning Mechanical Properties of Pseudopeptide Supramolecular Hydrogels by Graphene Doping.

Supramolecular hydrogels, obtained from small organic molecules, may be advantageous over polymeric ones for several applications, because these materials have some peculiar properties that differentiate them from the traditional polymeric hydrogels, such as elasticity, thixotropy, self-healing propensity, and biocompatibility. We report here the preparation of strong supramolecular pseudopeptide-based hydrogels that owe their strength to the introduction of graphene in the gelling mixture. These materials proved to be strong, stable, thermoreversible and elastic. The concentration of the gelator, the degree of graphene doping, and the nature of the trigger are crucial to get hydrogels with the desired properties, where a high storage modulus coexists with a good thixotropic behavior. Finally, NIH-3T3 cells were used to evaluate the cell response to the presence of the most promising hydrogels. The hydrogels biocompatibility remains good, if a small degree of graphene doping is introduced.

Oxazolidinone-containing pseudopeptides: Supramolecular materials, fibers, crystals, and gels.

The formation of fibers through self-assembly is of particular interest, as fibrous proteins (such as collagen, keratin, actin, and so on) are involved in intra- and extracellular functions. To understand aggregation phenomena, oligopeptides may be designed and prepared either to mimic or to interfere with these processes. In this article, we will demonstrate that the introduction of the 4-methyl-5-carboxy-oxazolidin-2-one (Oxd) moiety inside a peptide chain favors the formation of fiber-like materials organized either in ß-sheets or in supramolecular helices, provided that it is combined with other factors, like π-stacking interactions and intermolecular NH•••OC bonds. The presence of the Oxd moiety is essential for the material formation: when Oxd is replaced with Pro, only liquids or amorphous solids are obtained. Remarkably, some of these molecules are low-molecular-weight gelators, as they induce the formation of both organogel and hydrogels that have been used for several applications.

Synthesis and structure analysis of ferrocene-containing pseudopeptides.

Ferrocene with its aromaticity and facile redox properties is an attractive moiety to be incorporated into functional moieties. Medicinal applications of ferrocene are well known and ferrocene itself shows cytotoxic and antianemic properties. In this article, we will describe the synthesis and the structure analysis of two pseudopeptides containing a ferrocene moiety as N-terminal group. After purification, Fc-l-Phe-d-Oxd-OBn [l-Phel-phenylalanine; d-Oxd(4R,5S)-4-Methyl-5-carboxy-oxazolidin-2-one] appears as bright brown solid that spontaneously forms brown needles. The X-ray diffraction of the crystals shows the presence of strong π interactions between the ferrocenyl moiety and the phenyl rings, while no NH•••OC hydrogen bonds are formed. This result is confirmed by FT-IR and 1 H NMR analysis. In contrast, both FT-IR and 1 H NMR analysis suggest that Fc-(l-Phe-d-Oxd)2 -OBn forms a turn conformation stabilized by intramolecular NH•••OC hydrogen bonds in solution. Chiroptical spectroscopies (ECD and VCD) substantially confirmed the absence of a well-defined folded structure. The presence of the Fc moiety is responsible for specific ECD signals, one of which displayed pronounced temperature dependence and is directly related with the helicity assumed by the Fc core. Solid-state ECD spectra were recorded and rationalized on the basis of the X-ray geometry and quantum-mechanical calculations.

The effect of l-DOPA hydroxyl groups on the formation of supramolecular hydrogels.

Fmoc-l-DOPA-d-Oxd-OH was prepared starting from commercially available l-DOPA. Its gelation ability was tested by comparison with Fmoc-l-Tyr-d-Oxd-OH and Fmoc-l-Phe-d-Oxd-OH using ten different triggers. Among them, only GdL, CaCl2 and ZnCl2 form strong hydrogels with the three gelators. The analysis of the aerogels obtained by freeze drying the hydrogels show that the three gelators always induce the formation of dense networks, which strongly depend on the nature of the gelator. Rheological analysis of these samples demonstrates that stronger gels were obtained using the l-Tyr containing gelator, while the l-DOPA containing hydrogels were characterized by a storage modulus approximately one order of magnitude lower. Finally, the l-Phe containing gelators show a different trend with respect to the other samples depending on the trigger used. All the hydrogels show a thixotropic behaviour at the molecular level. These results indicate that hydrogel formation is sensitive to both the number of the hydroxyl moieties on the aromatic rings and trigger used.

Pseudopeptide-Based Hydrogels Trapping Methylene Blue and Eosin†Y.

We present herein the preparation of four different hydrogels based on the pseudopeptide gelator Fmoc-l-Phe-d-Oxd-OH (Fmoc=fluorenylmethyloxycarbonyl), either by changing the gelator concentration or adding graphene oxide (GO) to the water solution. The hydrogels have been analysed by rheological studies that demonstrated that pure hydrogels are slightly stronger compared to GO-loaded hydrogels. Then the hydrogels efficiency to trap the cationic methylene blue (MB) and anionic eosin Y (EY) dyes has been analyzed. MB is efficiently trapped by both the pure hydrogel and the GO-loaded hydrogel through π-π interactions and electrostatic interactions. In contrast, the removal of the anionic EY is achieved in less satisfactory yields, due to the unfavourable electrostatic interactions between the dye, the gelator and GO.

New azo-decorated N-pyrrolidinylthiazoles: synthesis, properties and an unexpected remote substituent effect transmission.

New 5-azo substituted thiazole derivatives have been obtained, under mild conditions and in good yields, by the reaction between 2-N-pyrrolidinylthiazole and a series of aryldiazonium salts bearing, mainly in the para position, groups with different electronic effects. The NMR spectra of the products show broad signals of the methylene groups in the alpha position to the pyrrolidinyl nitrogen, suggesting a hindered rotation around the C2-N bond, of which a double bond character was also evidenced by X-ray diffraction analyses. The free energies of activation for the rotational processes have been obtained from (1)H NMR experiments and computer simulations at different temperatures and provided good correlation with the σ constants of the substituents on the 'remote' benzene ring. This represents an unexpected and peculiar result since the restricted rotation around the C2-N bond was shown to be influenced by a substituent situated very far away. 2,4-Di-N-pyrrolidinylthiazole showed a much greater reactivity than 2-N-pyrrolidinylthiazole with aryldiazonium salts.

Hydrogelation Induced by Fmoc-Protected Peptidomimetics.

Four new low molecular weight hydrogelators (LMWGs) have been prepared in multigram scale and their attitude to form hydrogels has been tested. The gelation trigger is pH variation. The resulting gels have been characterized with several techniques: measurement of the melting points (T(gel)), transparency, gelation time, and viscoelastic properties, together with ECD analysis. Among them, Fmoc-L-Tyr-D-Oxd-OH 1 is an excellent gelator that leads to the preparation of strong, transparent, and viscoelastic gels, by pH variation. UV-visible analyses have demonstrated that the gels obtained with the LMWG 1 possess high transparency, with a transmittance up to 25.6% at a wavelength of 600 nm. Results of the amplitude sweep experiments showed that the elastic response component (G') was approximately an order of magnitude larger than the viscous component, indicating an elastic rather than viscous attitude of the gels, confirmed by the frequency independence of G' and G″ values, in the range from 0.1 to 100 rad·s(-1). The thermal behavior of gel obtained from Fmoc-L-Tyr-D-Oxd-OH 1 was characterized performing an "ad hoc" rheological temperature sweep experiment, that indicated that G' remained almost constant from 23 °C up to about 65 °C while G″ increased in the same temperature range. At higher temperatures, both G' and G″ values started to slightly decrease without displaying a crossover point.

Health System Response to the 2023 Floods in Emilia-Romagna, Italy: A Field Report.

In May 2023, the Italian region Emilia-Romagna was hit by intense rainfall, which caused extensive floods in densely populated areas. On May 4, 2023, a 12-month state of emergency was declared in the region with the activation of response and recovery plans. This field report provides an overview of the health response to the floods, paying particular attention to the measures put in place to ensure care for displaced populations and raising interesting points of discussion regarding the role of the health system during extreme weather events (EWEs). The considerations that emerge from this report underline the need for a primary care approach to disasters, especially when these occur in areas with a high prevalence of elderly resident population, and underscore the importance of integration of different levels of care.

Theoretical study of the absorption spectrum of a photoisomerizable iron complex.

We report the time-dependent density functional theory study of the absorption spectrum of an iron complex exhibiting photoisomerization properties. The role of the exchange-correlation functionals and, in particular, the effects of the inclusion of long-range corrections have been considered. The vertical transitions have been analyzed in terms of natural transition orbitals and have shown that the spectrum is dominated by ligand-to-metal charge-transfer transitions, an occurrence that could be promising for future applications of these complexes.

Low Molecular Weight Gelators Based on Functionalized l-Dopa Promote Organogels Formation.

We prepared the small pseudopeptide Lau-l-Dopa(OBn)2-d-Oxd-OBn (Lau = lauric acid; l-Dopa = l-3,4-dihydroxyphenylalanine; d-Oxd = (4R,5S)-4-methyl-5-carboxyl-oxazolidin-2-one; Bn = benzyl) through a number of coupling reactions between lauric acid, protected l-Dopa and d-Oxd with an excellent overall yield. The ability of the product to form supramolecular organogels has been tested with different organic solvents of increasing polarity and compared with the results obtained with the small pseudopeptide Fmoc-l-Dopa(OBn)2-d-Oxd-OBn. The mechanical and rheological properties of the organogels demonstrated solvent-dependent properties, with a storage modulus of 82 kPa for the ethanol organogel. Finally, to have a preliminary test of the organogels' ability to adsorb pollutants, we treated a sample of the ethanol organogel with an aqueous solution of Rhodamine B (RhB) for 24 h. The water solution slowly lost its pink color, which became trapped in the organogel.

Biocompatible and Light-Penetrating Hydrogels for Water Decontamination.

Solar light-activated photocatalyst nanoparticles (NPs) are promising environment-friendly low cost tools for water decontamination, but their dispersion in the environment must be minimized. Here, we propose the incorporation of TiO2-NPs (also in combination with graphene platelets) into highly biocompatible hydrogels as a promising approach for the production of photoactive materials for water treatment. We also propose a convenient fluorescence-based method to investigate the hydrogel photocatalytic activity in real time with a conventional fluorimeter. Kinetics analysis of the degradation profile of a target fluorescent model pollutant demonstrates that fast degradation occurs in the matrix bulk. Fluorescence anisotropy proved that small pollutant molecules diffuse freely in the hydrogel. Rheological and scanning electron microscopy characterization showed that the TiO2-NP incorporation does not significantly alter the hydrogel mechanical and morphological properties.

Peptide-Based Physical Gels Endowed with Thixotropic Behaviour.

Thixotropy is one of the oldest documented rheological phenomenon in colloid science and may be defined as an increase of viscosity in a state of rest and a decrease of viscosity when submitted to a constant shearing stress. This behavior has been exploited in recent years to prepare injectable hydrogels for application in drug delivery systems. Thixotropic hydrogels may be profitably used in the field of regenerative medicine, which promotes tissue healing after injuries and diseases, as the molten hydrogel may be injected by syringe and then self-adapts in the space inside the injection site and recovers the solid form. We will focus our attention on the preparation, properties, and some applications of biocompatible thixotropic hydrogels.

Thixotropic Peptide-Based Physical Hydrogels Applied to Three-Dimensional Cell Culture.

Pseudopeptides containing the d-Oxd or the d-pGlu [Oxd = (4R,5S)-4-methyl-5-carboxyl-oxazolidin-2-one, pGlu = pyroglutamic acid] moiety and selected amino acids were used as low-molecular-weight gelators to prepare strong and thixotropic hydrogels at physiological pH. The addition of calcium chloride to the gelator solutions induces the formation of insoluble salts that get organized in fibers at a pH close to the physiological one. Physical characterization of hydrogels was carried out by morphologic evaluation and rheological measurements and demonstrated that the analyzed hydrogels are thixotropic, as they have the capability to recover their gel-like behavior. As these hydrogels are easily injectable and may be used for regenerative medicine, they were biologically assessed by cell seeding and viability tests. Human gingival fibroblasts were embedded in 2% hydrogels; all of the hydrogels allow the growth of encapsulated cells with a very good viability. The gelator toxicity may be correlated with their tendency to self-assemble and is totally absent when the hydrogel is formed.

Form Matters: Stable Helical Foldamers Preferentially Target Human Monocytes and Granulocytes.

Some hybrid foldamers of various length, all containing the (4R,5S)-4-carboxy-5-methyloxazolidin-2-one (d-Oxd) moiety alternating with an l-amino acid (l-Val, l-Lys, or l-Ala), were prepared in order to study their preferred conformations and to evaluate their biological activity. Surprisingly, only the longer oligomers containing l-Ala fold into well-established helices, whereas all the other oligomers give partially unfolded turn structures. Nevertheless, they all show good biocompatibility, with no detrimental effects up to 64 µm. After equipping some selected foldamers with the fluorescent tag rhodamine B, a quantitative analysis was performed by dose- and time-response fluorescence-activated cell sorting (FACS) assays with human HeLa cells and primary blood lymphocytes, granulocytes, and monocytes. Among the cell types analyzed, the oligomers associated with monocytes and granulocytes with greatest efficacy, still visible after 24 h incubation. This effect is even more pronounced for foldamers that are able to form stable helices.

Factors Affecting the Stabilization of Polyproline II Helices in a Hydrophobic Environment.

Several parameters have a critical importance for the stabilization of either polyproline I (PPI) or polyproline II (PPII) helices in a hydrophobic environment. Among them, it was found out that the concentration is crucial as polyprolines at 3 mM concentration stably fold in PPII helices, that are organized in aggregates stable even after several days and are detectable by dynamic light scattering analysis. In more diluted concentration the same molecules stably fold in PPI helices, and no aggregates are found. In contrast, the introduction of a (4S,5R)-4-carboxy-5-methyloxazolidin-2-one (L-Oxd) moiety always inhibits the formation of the PPI helix, regardless of the L-Oxd position and the solution concentration.

Study on the interaction between gliadins and a coumarin as molecular model system of the gliadins-anthocyanidins complexes.

To clarify the conformational changes of gliadins (Glia) upon complexation with anthocyanidins (in particular cyanidin, Cya), the interaction of Glia with a coumarin derivative (3-ethoxycarbonylcoumarin, 3-EcC), having a benzocondensed structure similar to that of Cya, has been investigated by NMR, IR, and Raman spectroscopy under acidic and neutral conditions. Raman spectra showed that both molecules produce a similar effect on the Glia structure, i.e. an increase in the α-helix conformation and a decrease in ß-sheet and ß-turns content. In the presence of both molecules, this effect is more marked; the spectroscopic results showed that both Cya and 3-EcC interact with Glia and 3-EcC favors the complex formation with Glia. The results obtained in this study provide new insights into anthocyanidins-Glia interactions and may have relevance to human health, in the field of the attempts to modify gluten proteins to decrease allergen immunoreactivity.

On the antibacterial activity of roots of Capparis spinosa L.

A decoction of Capparis spinosa L. roots, widely used in the traditional folk medicine of southern Italy, was prepared and submitted to antibacterial activity tests, which showed an interesting bacteriostatic activity on the growth of Deinococcus radiophilus. Heterocyclic compounds were also recovered from the chloroformic extract of the roots.