Identification of Virulence Factors Involved in a Murine Model of Severe Achromobacter xylosoxidans Infection.

Achromobacter xylosoxidans (Ax) is an opportunistic pathogen and causative agent of numerous infections particularly in immunocompromised individuals with increasing prevalence in cystic fibrosis (CF). To date, investigations have focused on the clinical epidemiology and genomic comparisons of Ax isolates, yet little is known about disease pathology or the role that specific virulence factors play in tissue invasion or damage. Here, we model an acute Ax lung infection in immunocompetent C57BL/6 mice and immunocompromised CF mice, revealing a link between in vitro cytotoxicity and disease in an intact host. Mice were intratracheally challenged with sublethal doses of a cytotoxic (GN050) or invasive (GN008) strain of Ax. Bacterial burden, immune cell populations, and inflammatory markers in bronchoalveolar lavage fluid and lung homogenates were measured at different time points to assess disease severity. CF mice had a similar but delayed immune response toward both Ax strains compared to C57BL/6J mice. GN050 caused more severe disease and higher mortality which correlated with greater bacterial burden and increased proinflammatory responses in both mouse models. In agreement with the cytotoxicity of GN050 toward macrophages in vitro, mice challenged with GN050 had fewer macrophages. Mutants with transposon insertions in predicted virulence factors of GN050 showed that disease severity depended on the type III secretion system, Vi capsule, antisigma-E factor, and partially on the ArtA adhesin. The development of an acute infection model provides an essential tool to better understand the infectivity of diverse Ax isolates and enable improved identification of virulence factors important to bacterial persistence and disease.

Protein Based Amorphous Solid Dispersion: a Case Study Investigating Different Whey Proteins at High Drug Loading.

PURPOSE: Whey protein isolate (WPI) has previously been shown to be a promising new excipient for the development of amorphous solid dispersions (ASD) at a high drug loading of 50% (w/w). Whilst WPI is a protein mixture, comprising mainly the three proteins ß-lactoglobulin (BLG), α-lactalbumin (ALA), casein glycomacropeptides (CGMP), the individual contributions of these three proteins to the overall performance of whey protein based ASDs has still not been investigated. In addition, the limitations of the technology at even higher drug loadings (i.e., more than 50%) have not yet been explored. In this study, BLG, ALA, CGMP and WPI were each prepared as ASDs with the two poorly water-soluble drugs (Compound A and Compound B) at 50%, 60% and 70% drug loadings. METHODS: Solid state characterization, dissolution rate and physical stability of the obtained samples were analyzed. RESULTS: All the obtained samples were amorphous and showed faster dissolution rates compared to the respective pure crystalline drugs. However, the BLG based formulations-at least for Compound A-were outperforming the other ASDs in terms of stability, dissolution enhancement and solubility increase. CONCLUSION: Overall, the study confirmed that the investigated whey proteins showed their potential in developing ASDs even at high drug loadings of up to 70%.

Accessory mitral valve tissue: a differential diagnosis of an obstructive mass on the left ventricular outflow tract.

Accessory mitral valve tissue is a rare congenital cardiac anomaly that is typically discovered incidentally during echocardiographic evaluation prompted by an asymptomatic murmur. This pathology has characteristic echocardiographic elements and is usually associated with other CHD. The decision to perform surgical resection depends on factors such as the degree of obstruction, presence of symptoms, presence of other CHDs, and risk of thrombosis. The researchers hereby present a case of an asymptomatic paediatric patient with accessory mitral valve tissue that produced left ventricular outflow tract obstruction.

Modulating the spectroscopy and dynamics of a proton-transfer dye by functionalizing with phenyl groups.

Molecules undergoing excited-state proton transfer (ESPT) reactions are among the most interesting systems from spectroscopic and photophysical viewpoints. These molecules can be further functionalized with electron donating or accepting groups, inducing intramolecular charge transfer (ICT) events, which might be coupled to the ESIPT ones, conferring them with different spectroscopic and photophysical properties, which can be essential to implement the related materials in many key scientific and technological fields. Here, we report new benzimidazole derivatives that are functionalized with a phenyl group, 2-(5,10-diphenyl-1H-phenanthro[9,10-d]imidazol-2-yl)phenol (DP-HPPI), and its methylated equivalent, 2-(2-methoxyphenyl)-5,10-diphenyl-1H-phenanthro[9,10-d]imidazole (DP-MPPI). The results prove that these molecules in solutions undergo an ultrafast ICT (400-700 fs) reaction. Additionally, DP-HPPI also undergoes a reversible ESIPT process in dichloromethane. However, this is precluded in acetonitrile due to the involvement of intermolecular H-bonds in this solvent. These results provide key insights into the development of proton-transfer materials with bespoke spectral and photodynamical properties.

Timing of surgical repair of the stented aortic arch and coarctation in neonates.

We report a neonate with aortic arch hypoplasia and coarctation, in whom patency of the arterial duct could not be re-established and who was too ill to undergo primary surgical correction safely. This patient was treated in two stages: 1) angioplasty and stenting, 2) surgical correction. The safe period for surgical correction may be 2-3 weeks after angioplasty and stenting.

Inhibition of S.†aureus Infection of Human Umbilical Vein Endothelial Cells (HUVECs) by Trehalose- and Glucose-Functionalized Gold Nanoparticles.

Microbial adhesion to host cells represents the initial step in the infection process. Several methods have been explored to inhibit microbial adhesion including the use of glycopolymers based on mannose, galactose, sialic acid and glucose. These sugar receptors are, however, abundant in the body, and are not unique to bacteria. Trehalose, in contrast, is a unique disaccharide that is widely expressed by microbes. This carbohydrate has not yet been explored as an anti-adhesive agent. Herein, gold nanoparticles (AuNPs) coated with trehalose-based polymers were prepared and compared to glucose-functionalized AuNPs and examined for their ability to prevent binding to endothelial cells. Acting as anti-adhesive agents, trehalose-functionalized NPs decreased the binding of S. aureus to HUVECs, while outperforming the control NPs. Microscopy revealed that trehalose-coated NPs bound strongly to S. aureus compared to the controls. In conclusion, nanoparticles based on trehalose could be a non-toxic alternative to inhibit S. aureus infection.

Femto- to Millisecond Time-Resolved Photodynamics of a Double-Functionalized Push-Pull Organic Linker: Potential Candidate for Optoelectronically Active MOFs.

The design of improved organic linkers for the further engineering of smarter metal-organic framework (MOF) materials has become a paramount task for a wide number of material scientists. In this report, a luminescent double-functionalized push-pull (electron donor-acceptor) archetype organic molecule, dimethyl 4-amino-8-cyanonaphthalene-2,6-dicarboxylate (Me2CANADC), has been synthesized and characterized. The optical steady-state properties of Me2CANADC are strongly influenced by the surrounding environment as a direct consequence of its strong charge transfer (CT) character. The relaxation from its first electronically excited singlet state follows a double pathway: (1) on one side deactivating from its local excited (LE) state in the sub-picosecond or picosecond time domain, and (2) on the other side undergoing an ultrafast intramolecular charge transfer (ICT) reaction that is slowing down in viscous solvents. The deactivation to the ground state of these species with CT character is the origin of the Me2CANADC luminescence, and they present solvent-dependent lifetime values ranging from 8 to 18 ns. The slow photodynamics of Me2CANADC unveils the coexistence of a non-emissive triplet excited state and the formation of a long-lived charge separated state (2 µs). These observations highlight the promising optical properties of Me2CANADC linker, opening a window for the design of new functional MOFs with huge potential to be applied in the fields of luminescent sensing and optoelectronics.

Efficient Rare-Earth-Based Coordination Polymers as Green Photocatalysts for the Synthesis of Imines at Room Temperature.

Five new rare-earth coordination polymers (CPs) were designed in order to offer a remarkable platform that contains light-harvesting antennas and catalytic active centers to achieve solar-energy conversion as green alternatives in the synthesis of imines. These five new spirobifluorene-containing Ln-CPs, named [Er3(Hsfdc)3(sfdc)3(H2O)]· xH2O (RPF-30-Er), [Ln(Hsfdc)(sfdc)(EtOH)]·S (RPF-31-Ln, where Ln = La, Nd, and Sm and S = H2O or EtOH), and [Ho(Hsfdc)(sfdc)(H2O)] (RPF-32-Ho) (RPF = rare-earth polymeric framework and H2sfdc = 9,9'-spirobi[9 H-fluorene]-2,2'-dicarboxylic acid), have been solvothermally synthesized, and their structural features can be described as follows: (i) RPF-30-Er shows a 3D framework in which the inorganic trimers (secondary building units) are cross-linked by Hsfdc- and sfdc2- linkers displaying a pcu topology. (ii) The isostructural RPF-31-Ln series of materials, together with RPF-32-Ho, exhibit a 1D network of chains growing along the a axis with a ribbon-of-rings topology type. The photocatalytic activity of the RPF- n materials was tested in the oxidative coupling of amines using molecular oxygen and air as oxidizing agents under warm light. Among the materials investigated, RPF-31-Nd was chosen to further investigate the approach in the selectivity of different amine derivates.

Experimental and theoretical insights into the influence of electronic density on proton-transfer reactions.

We report on the excited-state behavior of proton-transfer phenanthroimidazole derivatives, such as HPPI and NMHPPI, in solutions using steady-state and femto- to nanosecond time-resolved fluorescence spectroscopies. Experimental observations are supported by theoretical calculations (TDDFT). In dichloromethane (DCM) and acetonitrile (ACN), two different paths are found for the excited-state intramolecular proton-transfer (ESIPT) reactions following two different channels. A fast and direct channel (ESIPT1) in 1-2.5 ps and a slower one (ESIPT2) in 12-15 ps, the latter being more influenced by the solvent viscosity (30 ps for HPPI and 20 ps for NMHPPI in triacetin (TAC) solutions). The slowing down of the ESIPT2 reaction is explained in terms of an intramolecular charge transfer (ICT) reaction coupled to a twisting motion to reach a more suitable conformation of the involved parts in the proton-transfer motion. The absence of OH/OD exchange effects in the ultrafast and slow proton-transfer dynamics suggests that the ESIPT reactions, which involve intramolecular and solvent coordinates, do not occur via tunneling. These results reveal new insights into the photobehavior of proton-transfer dyes, which might help in designing photosensors or lighting devices.

Influenza and respiratory syncytial virus in infants study (IRIS) of hospitalized and non-ill infants aged <1 year in four countries: study design and methods.

BACKGROUND: This multi-country prospective study of infants aged <1 year aims to assess the frequency of influenza virus and respiratory syncytial virus (RSV) infections associated with hospitalizations, to describe clinical features and antibody response to infection, and to examine predictors of very severe disease requiring intensive care. METHODS/DESIGN: We are enrolling a hospital-based cohort and a sample of non-ill infants in four countries (Albania, Jordan, Nicaragua, and the Philippines) using a common protocol. We are currently starting year 2 of a 2- to 3-year study and will enroll approximately 3,000 infants hospitalized for any acute illness (respiratory or non-respiratory) during periods of local influenza and/or RSV circulation. After informed consent and within 24 h of admission, we collect blood and respiratory specimens and conduct an interview to assess socio-demographic characteristics, medical history, and symptoms of acute illness (onset ≤10 days). Vital signs, interventions, and medications are documented daily through medical record abstraction. A follow-up health assessment and collection of convalescent blood occurs 3-5 weeks after enrollment. Influenza and RSV infection is confirmed by singleplex real time reverse transcriptase polymerase chain reaction (rRT-PCR) assays. Serologic conversion will be assessed comparing acute and convalescent sera using hemagglutination inhibition assay for influenza antibodies and enzyme-linked immunosorbent assay (ELISA) for RSV. Concurrent with hospital-based enrollment, respiratory specimens are also being collected (and tested by rRT-PCR) from approximately 1,400 non-ill infants aged <1 year during routine medical or preventive care. DISCUSSION: The Influenza and RSV in Infants Study (IRIS) promises to expand our knowledge of the frequency, clinical features, and antibody profiles of serious influenza and RSV disease among infants aged <1 year, quantify the proportion of infections that may be missed by traditional surveillance, and inform decisions about the potential value of existing and new vaccines and other prevention and treatment strategies.

The application of crystallization in the presence of additives to enable drug-in-capsule technology.

Crystallization in the presence of additives such as surfactants, polymers or impurities has been widely investigated in the pharmaceutical and chemical industries in order to change the crystal habit or to obtain a more desirable polymorph, affect crystal growth and influence dissolution. However, in this study, we investigated the concept of crystallization in the presence of surfactants in order to incorporate into the crystal lattice, a small amount (less than 1% w/w) of surfactant, sodium lauryl sulfate (SLS). The resulting crystals were further compared to crystals coated with SLS using a washing procedure; in order to assess whether either procedure generates improvements in the apparent solubility and dissolution of a poorly soluble drug so it can be filled directly into a capsule without the need of a complex formulation process.

Competitive Excimer Formation and Energy Transfer in Zr-Based Heterolinker Metal-Organic Frameworks.

The spectroscopy and dynamics of a series of Zr-based MOFs in dichloromethane suspension are reported. These Zr-NADC MOFs were constructed by using different mixtures of 2,6-naphthalenedicarboxylate (NDC) and 4-amino-2,6-naphthalenedicarboxylate (NADC) as organic linkers. The fraction of NADC relative to NDC in these heterolinker MOFs ranges from 2 to 35 %. The results indicate two competitive photoprocesses: NDC excimer formation and an energy transfer (ET) from excited NDC linkers to NADC linkers. Increasing the fraction of NADC linkers in the Zr-NADC nanostructure decreases the mean time constant of NDC excimer formation, while the NADC emission intensity experiences a drop at the highest fraction of this linker in the MOF. The first observation is explained by an increase in the energy-transfer probability between the two linkers, and the second by emission quenching in the NADC linkers due to ultrafast charge transfer assisted by the amino group. Femtosecond time-resolved emission studies showed that the ET process (recorded as decaying and rising components) from excited NDC to NADC takes place in 1.2 ps. Direct excitation of the NADC linkers (at 410 nm) shows a decaying, but not rising, component of 250-480 fs, which could reflect the formation of a nonemissive charge-separation state. The results show that by using MOFs having heterolinkers it is possible to trigger and tune excimer formation and ET processes.

Identification of new KLF1 and LU alleles during the resolution of Lutheran typing discrepancies.

BACKGROUND: The Lu(b) antigen is expressed on red blood cells (RBCs) of the majority of individuals in all populations. Its absence in transfused patients may lead to alloantibody production and mild-to-moderate transfusion reactions, and in pregnancies to mild hemolytic disease of the fetus and newborn. This report describes the results of discrepancy resolution between apparent LU*A/LU*B or LU*B/LU*B genotypes and apparent Lu(b-) or Lu(b+ weak) phenotypes in one Asian and 10 Caucasian blood donors. STUDY DESIGN AND METHODS: Whole blood samples were analyzed by molecular methods to resolve discrepancies between Lu(b-) phenotypes detected by serology and Lu(b+) phenotypes predicted by genotyping. RBC agglutination assays were performed with commercial and patient antisera by tube or gel column methods. Genotyping was performed on commercial arrays that target the LU*A/LU*B polymorphism at Position c.230. The discrepancies were resolved by sequencing of genomic DNA and in some cases by sequencing of cloned DNA fragments. RESULTS: Eleven new alleles with coding sequence variants were identified, seven in the KLF1 gene, which are presumed to act dominantly to silence LU expression, and four in the LU gene itself. The alleles are KLF1*114delC, KLF1*298T, KLF1*304C,484insC, KLF1*304C,1000del2, KLF1*621G, KLF1*948delC, KLF1*1040A,1045delT, LU*B(559T,711T,714T), LU*B(611A,638T), LU*B(1049del2ins3), and LU*B(1306T,1340T,1671T,1742T). CONCLUSION: Besides confirming common phenotypes and detecting rare antigen-negative phenotypes, the use of molecular methods in blood donor typing can prompt the identification of new alleles through discrepancy resolution.

Identification of six new RHCE variant alleles in individuals of diverse racial origin.

BACKGROUND: The introduction of molecular methods into routine blood typing is prompting the identification of new blood group alleles. Discrepancies between the results of genotyping and serology or chance events uncovered during genotyping prompted additional investigations, which revealed six new RHCE variant alleles. STUDY DESIGN AND METHODS: Samples from eight blood donors, two patients (one prenatal), and a patient's relative, all of diverse racial origin, were analyzed by standard serology methods, targeted genotyping arrays, DNA sequencing, and allele-specific polymerase chain reaction. RESULTS: Six new RHCE alleles were identified, namely, RHCE*cE84A, RHCE*ce202G, RHCE*ce307T, RHCE*Ce377G, RHCE*ce697G,712G,733G,744C, and RHCE*Ce733G. CONCLUSION: While implementation of new assays in commercial genotyping platforms to detect the polymorphisms reported here may not be justified given their apparent rarity, software interpretative algorithms may benefit from the identification of new alleles for a more accurate determination of genotypes and prediction of phenotypes.

Vehicle Systems and Excipients Used in Minipig Drug Development Studies.

Minipigs have been used for dermal drug development studies for decades, and they are currently more frequently considered as the second nonrodent species for pivotal nonclinical studies, in lieu of the dog or nonhuman primate, for compounds delivered via standard systemic routes of administration. Little is known about the tolerability of different excipients in minipigs; sharing knowledge of excipient tolerability and compositions previously used in nonclinical studies may avoid testing of inadequate formulations, thereby contributing to reduced animal usage. This article reviews vehicles employed in the Göttingen(®)minipig based on the combined experience from a number of pharmaceutical companies and contract research organizations. The review includes vehicles tolerated for single or multiple dosing by the Göttingen minipig, some of which are not appropriate for administration to other common nonrodent species (e.g., dogs). By presenting these data for dermal, oral, subcutaneous, and intravenous routes of administration, studies to qualify these vehicles in minipigs can be minimized or avoided. Additionally, investigators may more frequently consider using the minipig in place of higher species if the tolerability of a vehicle in the minipig is known.

Spectral and dynamical properties of a Zr-based MOF.

We report on the spectra and dynamics of a Zr-naphthalene dicarboxylic acid (Zr-NDC) MOF in different diluted solvent suspensions and in a concentrated tetrahydrofuran (THF) one. In a diluted diethyl ether (DE) suspension, we observed intraparticle excimer formation between neighboring naphthalene organic linkers, leading to a red-shifted broad band in the emission spectrum and to a dynamics composed of three components τ1 = 650 ps, τ2 = 3.7 ns and τ3 = 13.9 ns, assigned to the excimer photoproduction, monomer and excimer lifetimes, respectively. Furthermore, both absorption and emission spectra show a blue shift in more polar solvents characterized by the solvent polarity function f(ε,n). We also observed changes in the excimer formation time (490-840 ps) probably due to a variation in the MOF structural fluctuation induced by solvent filling. The global fluorescence quantum yield of these suspensions is around 0.30 ± 0.05. At higher concentrations of the MOF particles, we observed the absorption and emission signals of aggregates having an intercrystal excimer formation in ∼ 5 ps in a THF suspension, ∼ 100 times shorter than that observed in a diluted one. Our results give the spectral and dynamical properties of a Zr-NDC MOF in solvent suspensions, opening the way to further studies of these kinds of MOFs interacting with fluorescent dyes for possible photonic applications.

Photochemistry of Zr-based MOFs: ligand-to-cluster charge transfer, energy transfer and excimer formation, what else is there?

Understanding of the photocatalytic behaviour of Zr-based MOFs is fundamental for the improved design of new and more efficient photocatalysts. The present work describes steady-state and photodynamical studies on the behavior of two MOFs: Zr-2,6-naphthalene dicarboxylate (Zr-NDC) and Zr-4-amino-2,6-naphthalene dicarboxylate (Zr-NADC, 65% NDC/35% NADC) in dichloromethane (DCM) and N,N-dimethylformamide (DMF) suspensions. In the DMF suspension, the Zr-NDC MOF exhibits excimer formation in 280 ps due to the interactions between neighboring linkers. Using the femtosecond transient absorption (fs-TA) technique we have found that the Zr-NADC MOF exhibits a ligand-to-cluster charge transfer (LCCT) event in ∼170 and <100 fs in DMF and DCM, respectively. The Zr-NDC MOF shows similar LCCT in <100 fs in both solvents. The Zr-NADC MOF in DMF suspension shows an energy transfer (ET) from the excited NDC linkers to the NADC ones in 1.1 ps. Flash photolysis experiments demonstrate dominant radiative electron-hole (e--h+) recombination from two different trap states in 310 ns and 1.3 µs, and 32 ns and 1.0 µs in the photoexcited Zr-NADC and Zr-NDC MOFs in DMF, respectively. Excitation of the NDC linkers in the Zr-NADC and Zr-NDC MOFs in DCM allows for the characterization of the dynamics associated with the charge-separated state and the non-radiative recombination from the trap states (45 ns and 0.3 µs). Direct excitation of the NADC linkers (410 nm) in the Zr-NADC MOF in DCM suspension produces a radiative (e--h+) recombination in 3.5 µs. Further experiments in the presence of electron donor (N,N,N',N'-tetramethyl-p-phenylenediamine, TMPD) and acceptor (methyl viologen, MV2+) molecules corroborate the formation of charge separated states in these MOFs. These findings are relevant for understanding the photocatalytic and photonic behaviours of these MOFs.

From intra- to inter-molecular hydrogen bonds with the surroundings: steady-state and time-resolved behaviours.

We report on the photodynamics of 2-(2'-hydroxyphenyl)benzoxazole (HBO), compared to its amino derivatives, 6-amino-2-(2'-hydroxypheny)benzoxazole (6A-HBO) and 5-amino-2-(2'-hydroxypheny)benzoxazole (5A-HBO) in N,N-dimethylformamide (DMF) solutions. HBO at S0 shows a reversible deprotonation reaction leading to the production of anionic forms. However, for 6A-HBO and 5A-HBO, DMF containing KOH is necessary to produce the anions. Excited HBO in DMF exhibits intra- as well as inter-molecular proton transfer (ESIPT and ESPT) reactions. With excitation at 330 nm, we observed the open-enol, anti-enol and keto forms with different emission and lifetimes (620 ps, 1.5 ns, and 74 ps, respectively), while with the excitation at 433 nm, only the anionic species emission was detected (3.7 ns). Contrary to HBO, 6A-HBO and 5A-HBO do not exhibit any proton transfer process, and only the emissions of the open-enol charge-transferred forms (open-ECT) were observed, which are comparable to those of their methylated derivatives (6A-MBO and 5A-MBO). Femtosecond studies of 6A-MBO and 6A-HBO in DMF indicate that an intramolecular charge-transfer (ICT) reaction (∼80 fs) and solvent relaxation process (2 ps) take place at S1. Remarkably, the photoinduced breaking of the intramolecular hydrogen bond of 6A-HBO and the formation of an intermolecular hydrogen bond with DMF molecules occurs in 80 ps, while for 5A-HBO, this process occurs in less than 10 ps. In this study, we have demonstrated that the presence and position of the amino group in the HBO framework change both the S0 and S1 behaviours of the intramolecular H-bonds; a result which might be useful for the design and better understanding of supramolecular systems based on intra- and intermolecular H-bonds.

An abnormally slow proton transfer reaction in a simple HBO derivative due to ultrafast intramolecular-charge transfer events.

We report on the steady-state, picosecond and femtosecond time-resolved studies of a charge and proton transfer dye 6-amino-2-(2'-hydroxyphenyl)benzoxazole (6A-HBO) and its methylated derivative 6-amino-2-(2'-methoxyphenyl)benzoxazole (6A-MBO), in different solvents. With femtosecond resolution and comparison with the photobehaviour of 6A-MBO, we demonstrate for 6A-HBO in solution, the photoproduction of an intramolecular charge-transfer (ICT) process at S1 taking place in ∼140 fs or shorter, followed by solvent relaxation in the charge transferred species. The generated structure (syn-enol charge transfer conformer) experiences an excited-state intramolecular proton-transfer (ESIPT) reaction to produce a keto-type tautomer. This subsequent proton motion occurs in 1.2 ps (n-heptane), 14 ps (DCM) and 35 ps (MeOH). In MeOH, it is assisted by the solvent molecules and occurs through tunneling for which we got a large kinetic isotope effect (KIE) of about 13. For the 6A-DBO (deuterated sample in CD3OD) the global proton-transfer reaction takes place in 200 ps, showing a remarkable slow KIE regime. The slow ESIPT reaction in DCM (14 ps), not through tunnelling as it is not sensitive to OH/OD exchange, has however to overcome an energy barrier using intramolecular as well as solvent coordinates. The rich ESIPT dynamics of 6A-HBO in the used solutions is governed by an ICT reaction, triggered by the amino group, and it is solvent dependent. Thus, the charge injection to a 6A-HBO molecular frame makes the ICT species more stable, and the phenol group less acidic, slowing down the subsequent ESIPT reaction. Our findings bring new insights into the coupling between ICT and ESIPT reactions on the potential-energy surfaces of several barriers.

Direct observation of breaking of the intramolecular H-bond, and slowing down of the proton motion and tuning its mechanism in an HBO derivative.

We report on spectroscopic and photodynamical behaviours of 5-amino-2-(2'-hydroxyphenyl)benzoxazole (5A-HBO) in different solutions. The dye undergoes an ultrafast ICT reaction (<50 fs) (comparable to that observed for its methylated derivative, 5A-MBO), in agreement with the results of TD-DFT theoretical calculations (gas phase). Depending on the used solvent, the ICT reaction can be followed by a reversible/irreversible excited-state intramolecular proton transfer (ESIPT) reaction or by breaking of the intramolecular hydrogen bond (IHB). 5A-HBO in n-heptane solution exhibits an irreversible and slow (20 ps) ESIPT reaction, while that of the parent compound, HBO, takes place in less than 150 fs. Compared to excited HBO behaviour, theoretical calculations on 5A-HBO suggest a higher energy barrier (∼4 kcal mol(-1)) between the relaxed enol and keto tautomers, in addition to a less stabilization of the latter, which is in agreement with experiments in n-heptane. On the other hand, in dichloromethane, after the ICT reaction a subsequent and reversible proton motion occurs in an extraordinary slower regime (ns-time scale). No isotopic effect (OH/OD exchange) was observed in this solvent reflecting that the reversible ESIPT reaction evolves along the IHB and solvent coordinates. Using tetrahydrofurane and acetonitrile, we observed a breaking of the IHB due to specific intermolecular interactions with solvent molecules. This leads to the formation of open-enol forms, which undergo an ICT reaction as it occurs in 5A-MBO. These results bring new findings in the coupled ICT and ESIPT reactions. The photobehaviour of this new dye remarkably changes with the solvent nature, opening up the window for further research and possible applications in sensing polarity or H-bonding of media similar to that of the biological ones.