Supramolecular Recognition of Cytidine Phosphate in Nucleotides and RNA Sequences.

Supramolecular recognition of nucleotides would enable manipulating crucial biochemical pathways like transcription and translation directly and with high precision. Therefore, it offers great promise in medicinal applications, not least in treating cancer or viral infections. This work presents a universal supramolecular approach to target nucleoside phosphates in nucleotides and RNA. The artificial active site in new receptors simultaneously realizes several binding and sensing mechanisms: encapsulation of a nucleobase via dispersion and hydrogen bonding interactions, recognition of the phosphate residue, and a self-reporting feature-"turn-on" fluorescence. Key to the high selectivity is the conscious separation of phosphate- and nucleobase-binding sites by introducing specific spacers in the receptor structure. We have tuned the spacers to achieve high binding affinity and selectivity for cytidine 5' triphosphate coupled to a record 60-fold fluorescence enhancement. The resulting structures are also the first functional models of poly(rC)-binding protein coordinating specifically to C-rich RNA oligomers, e.g., the 5'-AUCCC(C/U) sequence present in poliovirus type 1 and the human transcriptome. The receptors bind to RNA in human ovarian cells A2780, causing strong cytotoxicity at 800 nM. The performance, self-reporting property, and tunability of our approach open up a promising and unique avenue for sequence-specific RNA binding in cells by using low-molecular-weight artificial receptors.

Tunable Macrocyclic Polyparaphenylene Nanolassos via Copper-Free Click Chemistry.

Deriving diverse compound libraries from a single substrate in high yields remains to be a challenge in cycloparaphenylene chemistry. In here, a strategy for the late-stage functionalization of shape-persistent alkyne-containing cycloparaphenylene has been explored using readily available azides. The copper-free [3+2]azide-alkyne cycloaddition provided high yields (>90 %) in a single reaction step. Systematic variation of the azides from electron-rich to -deficient shines light on how peripheral substitution influences the characteristics of the resulting adducts. We find that among the most affected properties are the molecular shape, the oxidation potential, excited state features, and affinities towards different fullerenes. Joint experimental and theoretical results are presented including calculations with the state-of-the-art, artificial intelligence-enhanced quantum mechanical method 1 (AIQM1).

Well-separated water-soluble carbon dots via gradient chromatography.

Carbon dots (CDs) are strongly fluorescent advanced materials that are promising for applications in bio-imaging, sensors or luminescent displays. One of the most-widely used class of CDs is synthesized via an aqueous, bottom-up technique starting from citric acid (CA) and an amino-precursor. Very high fluorescence quantum yields (QY) are reported for the resulting CDs. The as-synthesized raw suspensions, however, are crude mixtures of many components: bare carbon cores, carbon cores functionalized with fluorophores, freely floating molecular fluorophores, and several other by-products. In this study, we synthesized CDs from CA and amino acid cysteine (Cys) hydrothermally and demonstrate a complete separation of all components by means of two step gradient chromatography. In the first step, the separation was carried out on a normal-pressure preparative silica-gel column to get sufficient amounts of material to investigate structure and optical properties of the collected fractions. This preparative gradient elution method enabled us to separate moderately-fluorescent CDs from freely floating molecular fluorophores, polymeric fluorophores and CDs with built-in fluorophores. Here, we evidenced that amorphous CDs co-exist with crystalline CDs in one and the same suspension and showed that the amount of crystalline CDs increases with the synthesis temperature. In the second step, we turned to high performance liquid chromatography (HPLC) to further improve and optimize the efficiency of purification and automate it. Via HPLC, we were able to well-separate of up to six components. Within this work, we laid the foundation for CD purification with the highest possible purity for aqueous, bottom-up synthesized CDs and quantified the true quantum yield of CDs.

Purification and structural elucidation of carbon dots by column chromatography.

Carbon dots (CDs) are an astonishing class of fluorescent materials with many applications in bioimaging, drug delivery, photovoltaics and photocatalysis due to their outstanding luminescence properties and low toxicity. However, the internal CD structure of bottom-up synthesized CDs is still the subject of considerable debate. Unambiguous analysis of the internal CD composition is hampered by the fact that reaction products usually contain mixtures of several CD fractions as well as molecular intermediate and side products. Therefore, purification and careful separation of the various CD fractions is vital for structural analysis and isolation of pure CDs possessing optimized optical properties. In this study, CD solutions were synthesized from citric acid and cysteine via a one-pot hydrothermal synthesis. A simple column chromatography unit was used to systematically study the influence of the molar precursor ratios and synthesis conditions (temperature, reaction time) on the CD solution composition. By investigating the structural and optical properties of the chromatographically separated fractions, three different fluorescent species could be identified. Freely floating molecular fluorophores left the column first, followed by highly fluorescent CDs with fluorophores bound to the carbon core, finally followed by low-fluorescent carbon particles without fluorophores. We demonstrate that the CD solution composition and the internal structure of the individual fluorescent components can be clarified via chromatographic separation. This information can be further applied to isolate pure CDs with optimized optical properties.

Hierarchical organization of perylene bisimides and polyoxometalates for photo-assisted water oxidation.

The oxygen in Earth's atmosphere is there primarily because of water oxidation performed by photosynthetic organisms using solar light and one specialized protein complex, photosystem II (PSII). High-resolution imaging of the PSII 'core' complex shows the ideal co-localization of multi-chromophore light-harvesting antennas with the functional reaction centre. Man-made systems are still far from replicating the complexity of PSII, as the majority of PSII mimetics have been limited to photocatalytic dyads based on a 1:1 ratio of a light absorber, generally a Ru-polypyridine complex, with a water oxidation catalyst. Here we report the self-assembly of multi-perylene-bisimide chromophores (PBI) shaped to function by interaction with a polyoxometalate water-oxidation catalyst (Ru4POM). The resulting [PBI]5Ru4POM complex shows a robust amphiphilic structure and dynamic aggregation into large two-dimensional paracrystalline domains, a redshifted light-harvesting efficiency of >40% and favourable exciton accumulation, with a peak quantum efficiency using 'green' photons (λ > 500 nm). The modularity of the building blocks and the simplicity of the non-covalent chemistry offer opportunities for innovation in artificial photosynthesis.

The energy-transfer-enabled biocompatible disulfide-ene reaction.

Sulfur-containing molecules participate in many essential biological processes. Of utmost importance is the methylthioether moiety, present in the proteinogenic amino acid methionine and installed in tRNA by radical-S-adenosylmethionine methylthiotransferases. Although the thiol-ene reaction for carbon-sulfur bond formation has found widespread applications in materials or medicinal science, a biocompatible chemo- and regioselective hydrothiolation of unactivated alkenes and alkynes remains elusive. Here, we describe the design of a general chemoselective anti-Markovnikov hydroalkyl/aryl thiolation of alkenes and alkynes-also allowing the biologically important hydromethylthiolation-by triplet-triplet energy transfer activation of disulfides. This fast disulfide-ene reaction shows extraordinary functional group tolerance and biocompatibility. Transient absorption spectroscopy was used to study the sensitization process in detail. The hereby gained mechanistic insights were successfully employed for optimization of the catalytic system. This photosensitized transformation should stimulate bioimaging applications and carbon-sulfur bond-forming late-stage functionalization chemistry, especially in the context of metabolic labelling.

Bulbous gold-carbon nanodot hybrid nanoclusters for cancer therapy.

Assemblies of inorganic nanoparticles and carbon nanodots have emerged as promising candidates for hybrid materials in biomedical applications. In this work, the formation and properties of gold nanoparticles synthesized with the aid of carbon nanodots (CND) as reducing/stabilizing agents was investigated. Through careful modification of the reaction conditions, such as precursor concentrations and temperature, the size and shape of the particles can be controlled. In general, CNDs provide reductive sites at which gold seeds can be formed. As the gold nanoparticles grow, the CNDs form a polar solubilizing shell in the polar aqueous environment, yielding bulbous Au@pCND nanoclusters. In fact, charge-transfer interactions between the pCND shell and the Au core are implicit, as confirmed by steady state and time resolved absorption and fluorescence spectroscopies. Finally, this work demonstrated via in cell internalization tests that the size and the shape of the Au@pCND nanoclusters play a crucial role for the cancer cell toxicity. Also the Au@pCND prove to be good candidates as sensitizers in cancer radio-therapy.

Carbon nanotubes dispersed in aqueous solution by ruthenium(ii) polypyridyl complexes.

Cationic ruthenium(ii) polypyridyl complexes with appended pyrene groups have been synthesized and used to disperse single-walled carbon nanotubes (SWCNT) in aqueous solutions. To this end, planar pyrene groups enable association by means of π-stacking onto carbon nanotubes and, in turn, the attachment of the cationic ruthenium complexes. Importantly, the ionic nature of the ruthenium complexes allows the formation of stable dispersions featuring individualized SWCNTs in water as confirmed in a number of spectroscopic and microscopic assays. In addition, steady-state photoluminescence spectroscopy was used to probe the excited state interactions between the ruthenium complexes and SWCNTs. These studies show that the photoluminescence of both, that is, of the ruthenium complexes and of SWCNTs, are quenched when they interact with each other. Pump-probe transient absorption experiments were performed to shed light onto the nature of the photoluminescence quenching, showing carbon nanotube-based bands with picosecond lifetimes, but no new bands which could be unambigously assigned to photoinduced charge transfer process. Thus, from the spectroscopic data, we conclude that quenching of the photoluminescence of the ruthenium complexes is due to energy transfer to proximal SWCNTs.

Probing Supramolecular Interactions between a Crown Ether Appended Zinc Phthalocyanine and an Ammonium Group Appended to a C60 Derivative.

Self-assembly driven by crown ether complexation of zinc phthalocyanines equipped with one 18-crown-6 moiety and fullerenes bearing an ammonium head group afforded a novel donor-acceptor hybrid. In reference experiments, fullerenes containing a Boc-protected amine functionality have been probed. The circumvention of zinc phthalocyanine aggregation is important for the self-assembly, which required the addition of pyridine. From absorption and fluorescence titration assays, which provided sound and unambiguous evidence for mutual interactions between the electron donor and the electron acceptor within the hybrids, association constants in the order of 8.0×105 m-1 have been derived. The aforementioned is based on 1:1 stoichiometries, which have been independently confirmed by Job's plot measurements. In the excited state, which has been examined by transient absorption experiments, intermolecular charge separation evolves from the photoexcited zinc phthalocyanine to the fullerene subunit and leads to short-lived charge-separated states. Interestingly, photoexcitation of zinc phthalocyanine dimers/aggregates can also be followed by an intermolecular charge separation between vicinal phthalocyanines. These multicomponent supramolecular ensembles have also been shown by in-depth electrospray ionization mass spectrometry (ESI-MS) studies, giving rise to the formation and detection of a variety of non-covalently linked species.

Unveiling the nature of supramolecular crown ether-C60 interactions.

A series of exTTF-(crown ether)2 receptors, designed to host C60, has been prepared. The size of the crown ether and the nature of the heteroatoms have been systematically changed to fine tune the association constants. Electrochemical measurements and transient absorption spectroscopy assisted in corroborating charge transfer in the ground state and in the excited state, leading to the formation of radical ion pairs featuring lifetimes in the range from 12 to 21 ps. To rationalize the nature of the exTTF-(crown ether)2·C60 stabilizing interactions, theoretical calculations have been carried out, suggesting a synergetic interplay of donor-acceptor, π-π, n-π and CH···π interactions, which is the basis for the affinity of our novel receptors towards C60.

Mechanistic aspects of the radiation-chemical reduction of graphene oxide to graphene-like materials.

PURPOSE: The aim of the work was to investigate mechanistic details of the preparation of graphene-like materials (GLM) via reduction of graphene oxide (GO) in aqueous dispersions by electron beam (EB) generated reducing free radicals. MATERIALS AND METHODS: A 10 MeV linear accelerator was employed to irradiate aqueous GO dispersions at ambient temperatures. The kinetics of GO reduction was followed using UV-Vis spectroscopy. The resulting GLM were characterized by X-ray photoelectron spectroscopy (XPS), Transmission electron microscopy (TEM), Raman spectroscopy and conductivity measurements. RESULTS: The reduction of GO could be afforded with high efficiency within minutes at room temperature via the reaction of GO with reducing radicals generated by EB irradiation. The detailed investigation of the reduction mechanism allowed a selection of the best reducing free radicals in terms of both their efficiency and environmental impact of their precursors and final products. CONCLUSIONS: The EB-treatment of aqueous GO dispersions is a highly efficient, environmentally friendly, cost-effective and easily up-scalable method for the preparation of GLM. The efficiency of the new reduction approach is comparable with the best existing methods.

Chromoendoscopy in magnetically guided capsule endoscopy.

BACKGROUND: Diagnosis of intestinal metaplasia and dysplasia via conventional endoscopy is characterized by low interobserver agreement and poor correlation with histopathologic findings. Chromoendoscopy significantly enhances the visibility of mucosa irregularities, like metaplasia and dysplasia mucosa. Magnetically guided capsule endoscopy (MGCE) offers an alternative technology for upper GI examination. We expect the difficulties of diagnosis of neoplasm in conventional endoscopy to transfer to MGCE. Thus, we aim to chart a path for the application of chromoendoscopy on MGCE via an ex-vivo animal study. METHODS: We propose a modified preparation protocol which adds a staining step to the existing MGCE preparation protocol. An optimal staining concentration is quantitatively determined for different stain types and pathologies. To that end 190 pig stomach tissue samples with and without lesion imitations were stained with different dye concentrations. Quantitative visual criteria are introduced to measure the quality of the staining with respect to mucosa and lesion visibility. Thusly determined optimal concentrations are tested in an ex-vivo pig stomach experiment under magnetic guidance of an endoscopic capsule with the modified protocol. RESULTS: We found that the proposed protocol modification does not impact the visibility in the stomach or steerability of the endoscopy capsule. An average optimal staining concentration for the proposed protocol was found at 0.4% for Methylene blue and Indigo carmine. The lesion visibility is improved using the previously obtained optimal dye concentration. CONCLUSIONS: We conclude that chromoendoscopy may be applied in MGCE and improves mucosa and lesion visibility. Systematic evaluation provides important information on appropriate staining concentration. However, further animal and human in-vivo studies are necessary.

Quantum-dot-sensitized solar cells: understanding linker molecules through theory and experiment.

We have investigated the role of linker molecules in quantum-dot-sensitized solar cells (QDSSCs) using density-functional theory (DFT) and experiments. Linkers not only govern the number of attached QDs but also influence charge separation, recombination, and transport. Understanding their behavior is therefore not straightforward. DFT calculations show that mercaptopropionic acid (MPA) and cysteine (Cys) exhibit characteristic binding configurations on TiO(2) surfaces. This information is used to optimize the cell assembly process, yielding Cys-based cells that significantly outperform MPA cells, and reach power conversion efficiencies (PCE) as high as 2.7% under AM 1.5 illumination. Importantly, the structural information from theory also helps understand the cause for this improved performance.

Glycophthalocyanines as photosensitizers for triggering mitotic catastrophe and apoptosis in cancer cells.

Photodynamic therapy (PDT) is a treatment modality for different forms of cancer based on the combination of light, molecular oxygen, and a photosensitizer (PS) compound. When activated by light, the PS generates reactive oxygen species leading to tumor destruction. Phthalocyanines are compounds that have already shown to be efficient PSs for PDT. Several examples of carbohydrate substituted phthalocyanines have been reported, assuming that the presence of carbohydrate moieties could improve their tumor selectivity. This work describes the photoeffects of symmetric and asymmetric phthalocyanines with D-galactose (so-called GPh1, GPh2, and GPh3) on HeLa carcinoma cells and their involvement in cell death. Photophysical properties and in vitro photodynamic activities for the compounds considered revealed that the asymmetric glycophthalocyanine GPh3 is very efficient and selective, producing higher photocytotoxicity on cancer cells than in nonmalignat HaCaT. The cell toxiticy after PDT treatment was dependent upon light exposure level and GPh3 concentration. GPh3 causes cell cycle arrest at the metaphase stage leading to multiple spindle poles, mitotic catastrophe, followed by apoptosis in cancer cells. These effects were partially negated by the pancaspase inhibitor Z-VAD-FMK. Together, these results indicate that GPh3 is an excellent candidate drug for PDT, able to induce selective tumor cell death.

A new exTTF-crown ether platform to associate fullerenes: cooperative n-π and π-π effects.

A new and readily available exTTF-bis(crown ether), 1, efficiently recognizes C60 as well as C70 by means of cooperative π-π and n-π interactions. The geometrical (concave-convex) and electronic (donor-acceptor) complementarity accounts on one hand for remarkable binding strengths, with association constants reaching 10(7) M(-1) in benzonitrile, and on the other hand for lifetimes of the photogenerated radical ion pair state on the order of 45 ps.

Photoinduced energy transfer processes within dyads of metallophthalocyanines compactly fused to a ruthenium(II) polypyridine chromophore.

An unsymmetric, peripherally octasubstituted phthalocyanine (Pc) 1, which contains a combination of dipyrido[3,2-f:2',3'-h] quinoxaline and 3,5-di-tert-butylphenoxy substituents, has been obtained via a statistical condensation reaction of two corresponding phthalonitriles. Synthetic procedures for the selective metalation of the macrocyclic cavity and the periphery of 1 were developed, leading to the preparation of the key precursor metallophthalocyanines 3-5 in good yields. Two different strategies were applied to the synthesis of compact dyads MPc-Ru(II) 6-8 (M = Mg(II), Co(II), Zn(II)). Intramolecular electronic interactions in these dyads were studied by absorption, emission, and transient absorption spectroscopy. Upon photoexcitation, these dyads exhibit efficient intramolecular energy transfer from the Ru(II) chromophore to the MPc moiety.

Electron transfer in Me-blocked heterodimeric alpha,gamma-peptide nanotubular donor-acceptor hybrids.

Bio-inspired cyclopeptidic heterodimers built on beta-sheet-like hydrogen-bonding networks and bearing photoactive and electroactive chromophores on the outer surface have been prepared. Different cross-strand pairwise relationships between the side chains of the cyclic alpha,gamma-peptides afford the heterodimers as three nonequivalent dimeric species. Steady-state and time-resolved spectroscopies clearly show an electron transfer process from pi-extended tetrathiafulvalene, covalently attached to one of the cyclopeptides, to photoexcited [60]fullerene, located on the complementary cyclopeptide. The charge-separated state was stabilized for up to 1 micros before recombining and repopulating the ground state. Our current example shows that cyclopeptidic templates can be successfully used to form light-harvesting/light-converting hybrid ensembles with a distinctive organization of donor and acceptor units able to act as efficient artificial photosystems.

Fullerene polypyridine ligands: synthesis, ruthenium complexes, and electrochemical and photophysical properties.

Fullerene coordination ligands bearing one bipyridine or terpyridine unit were synthesized, and their coordination to ruthenium(II) formed linear rod-like donor-acceptor systems. Steady-state fluorescence of [Ru(bpy)(2)(bpy-C(60))](2+) showed a rapid solvent-dependent, intramolecular quenching of the ruthenium(II) MLCT excited state. Time-resolved flash photolysis in CH(3)CN revealed characteristic transient absorption changes that have been ascribed to the formation of the C(60) triplet state, suggesting that photoexcitation of [Ru(bpy)(2)(bpy-C(60))](2+) results in a rapid intramolecular transduction of triplet excited state energy. The electrochemical studies on both [Ru(bpy)(2)(bpy-C(60))](2+) and [Ru(tpy)(tpy-C(60))](2+) indicated electronic coupling between the metal center and the fullerene core.

A helical peptide receptor for [60]fullerene.

Two terminally blocked nonapeptides, each made up of six Aib residues, a Gly spacer and two L-Tyr residues in positions 2 and 8 (these are substituted in the side chain with either ferrocenoyl or methyl moieties), have been synthesized by solution methods and fully characterized. FT-IR absorption and two-dimensional NMR analyses indicate that a 3(10)-helical conformation is adopted by these rigid peptides in structure-supporting solvents. An X-ray diffraction investigation shows that the bis-L-Tyr(Me) nonapeptide in the crystal state is folded in a regular right-handed 3(10)-helical structure. As five amino acid units separate the two substituted L-Tyr residues in the peptide sequence, the two side chain moieties will-in solution-face each other after two complete turns of the ternary helix. By carrying out a detailed photophysical analysis, we have demonstrated that the electron-rich, hydrophobic and wide cavity generated by the nonapeptide template with two ferrocenoyloxybenzyl walls is able to host [60]fullerene. Further evidence for this superstructure has been provided in the gas phase by a mass spectrometric investigation.