Modulating the G-Quadruplex and Duplex DNA Binding by Controlling the Charge of Fluorescent Molecules.

Two fluorescent and non-toxic spirobifluorene molecules bearing either positive (Spiro-NMe3) or negative (Spiro-SO3) charged moieties attached to the same aromatic structure have been investigated as binders for DNA. The novel Spiro-NMe3 containing four alkylammonium substituents interacts with G-quadruplex (G4) DNA structures and shows preference for G4s over duplex by means of FRET melting and fluorescence experiments. The interaction is governed by the charged substituents of the ligands as deduced from the lower binding of the sulfonate analogue (Spiro-SO3). On the contrary, Spiro-SO3 exhibits higher binding affinity to duplex DNA structure than to G4. Both molecules show a moderate quenching of the fluorescence upon DNA binding. The confocal microscopy evaluation shows the internalization of both molecules in HeLa cells and their lysosomal accumulation.

Cationic Solution and Solid-State Emitters - Robust Imaging Agents for Cells, Bacteria, and Protists.

A library of eight different cationic emitters with emission properties in solution and in solid-state (solution and solid-state emitters - SSSE) is presented. These compounds, bearing either ammonium or pyridinium groups, have been investigated regarding their photophysical properties as well as their potential application in biological imaging. Besides high quantum yields as well as a high degree of stability during the imaging process, it was additionally revealed that a broad range of biological targets can be addressed, such as different bacterial strains, human cells as well as protists. The reported SSSE approach employing the mentioned robust emitters for biological imaging, will contribute to a rapid and facile way to design and apply affordable emitters with outstanding properties. Additionally, these emitters will overcome the drawbacks of classical luminophores and agents featuring well-known aggregation-induced emission (AIE) or aggregation-caused quenching (ACQ) properties.

Regioselectivity Control in Spirobifluorene Nitration under Mild Conditions: Explaining the Crivello's Reagent Mechanism.

The regioselective nitration of 9,9'-spirobifluorene under mild conditions is reported for the first time by operating under Menke's and Crivello's conditions. The optimized protocol allows obtaining 2-nitro and 2,2'-dinitro-9,9'-spirobifluorene in yields of 79 and 95% and, for the first time, 2,2',7-trinitro-9,9'-spirobifluorene with 66% yield. Besides, the role of dinitrate salt in Crivello's protocol has been now clarified, which opens novel scenarios in the preparation of functional materials.

Deoxyestrone-based lipofection agents with solution- and solid-state emission properties.

In this contribution, three deoxyestrone-based emissive lipofection agents are reported. Because of a centrally incorporated terephthalonitrile motif, these ligands can be classified as solution and solid-state emitters (SSSEs). With the attachment of tobramycin, these amphiphilic structures are able to form lipoplexes, mediating gene transfection of HeLa and HEK 293T cells.

Biofortification of cow milk through dietary supplementation with sunflower oil: fatty acid profile, atherogenicity, and thrombogenic index.

The present study aimed to assess the effects of replacing the starchy ingredients of concentrate by increasing the levels of sunflower oil on the production, composition, fatty acid profile, and evaluate the atherogenicity and thrombogenic index of Jersey cow's milk. Eight Jersey cows were arranged in a double Latin square and distributed in treatments consisting of supplementation with increasing levels of sunflower oil replacing the corn grain and wheat bran of concentrate, including the following: T0 (control diet), without sunflower oil and with 38 g ether extract (EE)/kg dry matter (DM); T1 = 65 g EE/kg DM; T2 = 86 g EE/kg DM; and T3 = 110 g EE/kg DM. The daily milk production was measured, and the corrected milk production was calculated. Milk samples were analyzed by infrared spectroscopy to determine fat, protein, lactose, and total solids, whereas the lipid profile was assessed by gas chromatography. Milk production, energy-corrected milk production, fat content, daily fat production, lactose, and total solids were not affected by the treatments. Protein, lactose, and total solids concentrations decreased. Short-, medium-, and odd-chain fatty acids decreased with an increase in sunflower oil levels. Conversely, linear increases in long-chain, monounsaturated, and polyunsaturated fatty acid concentrations were observed. There were significant increases in stearic and elaidic acids and conjugated linoleic acid isomers, especially in vaccenic and rumenic acids. There was a positive effect on the milk atherogenicity, thrombogenicity, and nutraceutical indices. Dietary supplementation with sunflower oil changes the milk FA profile, decreases the atherogenicity and the thrombogenicity indices, and improve the nutraceutical index up to the addition of 86 g EE/kg DM de sunflower oil in the diets of Jersey cows.

Phosphorescence Induction by Host-Guest Complexation with Cyclodextrins - The Role of Regioisomerism and Affinity.

We present an in-depth investigation of cyclodextrin complexes with guest compounds featuring complexation-induced room temperature phosphorescence (RTP) in aqueous solution. Very interestingly, only the complexed regioisomers bearing lateral substituents on meta-position show RTP, whereas the stronger host-guest systems with para-substituted dyes show no RTP features. The reported systems were investigated regarding their complexation behavior in water using isothermal titration calorimetry and mass spectrometry. In the case of γ-CD very strong 1 : 1 inclusion complexes (Ka up to 5.13×105  M-1 ) were unexpectedly observed. It was found that not only a strong binding to the cyclodextrin cavity is needed to restrict motion, inducing the emission, but also the conformation inside the cavity plays a pivotal role - as supported by an extensive NMR study and MD simulations.

Bypassing the statistical limit of singlet generation in sensitized upconversion using fluorinated conjugated systems.

The photon upconversion based on triplet-triplet annihilation (TTA) is a mechanism that converts the absorbed low-energy electromagnetic radiation into higher energy photons also at extremely low excitation intensities, but its use in actual technologies is still hindered by the limited availability of efficient annihilator moieties. We present here the results obtained by the synthesis and application of two new fluorinated chromophores based on phenazine and acridine structures, respectively. Both compounds show upconverted emission demonstrating their ability as TTA annihilator. More interesting, the acridine-based chromophore shows an excellent TTA yield that overcomes the one of some of best model systems. By correlating the experimental data and the quantum mechanical modeling of the investigated compound, we propose an alternative efficient pathway for the generation of the upconverted emissive states involving the peculiar high-energy triplet levels of the dye, thus suggesting a new development strategy for TTA annihilators based on the fine tuning of their high-energy excited states properties.

Mapping the Regioisomeric Space and Visible Color Range of Purely Organic Dual Emitters with Ultralong Phosphorescence Components: From Violet to Red Towards Pure White Light.

We mapped the entire visible range of the electromagnetic spectrum and achieved white light emission (CIE: 0.31, 0.34) by combining the intrinsic ns-fluorescence with ultralong ms-phosphorescence from purely organic dual emitters. We realized small molecular materials showing high photoluminescence quantum yields (ΦL ) in the solid state at room temperature, achieved by active exploration of the regioisomeric substitution space. Chromophore stacking-supported stabilization of triplet excitons with assistance from enhanced intersystem crossing channels in the crystalline state played the primary role for the ultra-long phosphorescence. This strategy covers the entire visible spectrum, based on organic phosphorescent emitters with versatile regioisomeric substitution patterns, and provides a single molecular source of white light with long lifetime (up to 163.5 ms) for the phosphorescent component, and high overall photoluminescence quantum yields (up to ΦL =20 %).

From Blue to Green: Fine-Tuning of Photoluminescence and Electrochemiluminescence in Bifunctional Organic Dyes.

We describe the synthesis, computational analysis, photophysics, electrochemistry and electrochemiluminescence (ECL) of a series of compounds formed of two triphenylamines linked by a fluorene or spirobifluorene bridge. The phenylamine moieties were modified at the para-position of the two external rings by electron-withdrawing or electron-donating substituents. These modifications allowed for fine-tuning of the photoluminescence (PL) and ECL emission from blue to green, with an overall wavelength span of 73 (PL) and 67 (ECL) nm, respectively. For all compounds, we observed a very high PL quantum yield (79-89%) and formation of stable radical ions. The ECL properties were investigated by direct annihilation of the electrogenerated radical anion and radical cation. The radical-ion annihilation process is very efficient and causes an intense greenish-blue ECL emission, easily observable even by naked eye, with quantum yield higher than the standard 9,10-diphenylanthracene. The ECL spectra show one single band that almost matches the PL band. Because the energy of the annihilation reaction is higher than that required to form the singlet excited state, the S-route is considered the favored pathway followed by the ECL process in these molecules. All these features point to this type of molecular system as promising for ECL applications.

Fluorescence quenching in ß-cyclodextrin vesicles: membrane confinement and host-guest interactions.

Fluorescent ß-cyclodextrin vesicles (ß-CDV) that display host cavities available for host-guest interactions at the vesicle surface were prepared by incorporation of the hydrophobic spirobifluorene-based dye 1 into the membrane of unilamellar vesicles. Fluorescence quenching of dye 1 was observed in the presence of different quenchers. Methyl viologen 2 does not quench dye 1 because it does not bind to ß-CDV. 4-Nitrophenol 3 and 4-nitrophenol covalently connected to adamantane 4 quench the fluorescence of dye 1 in neutral solution, but by different mechanisms according to lifetime measurements. The quenching efficiency of 3 is pH dependent due to the presence of the phenolate form. Competition experiments with excess host and guest showed that 3 is likely to diffuse in and out of the membrane, while 4 forms an inclusion complex with ß-CDV leading to close contact and efficient quenching. Our findings confirm that this dynamic supramolecular system is a versatile model to investigate quenching and recognition processes in bilayer membranes.

Efficient luminescence from fluorene- and spirobifluorene-based lanthanide complexes upon near-visible irradiation.

We describe herein the synthesis and photophysical characterization of new lanthanide complexes that consist of a (9,9-dimethylfluoren-2-yl)-2-oxoethyl or a (9,9'-spirobifluoren-2-yl)-2-oxoethyl unit as the antenna, covalently linked to a 1,4,7,10-tetraazacyclododecane-1,4,7-triacetic acid (DO3A) unit as the Ln(3+) (Gd(3+), Eu(3+), Sm(3+), Tb(3+), Dy(3+)) coordination site. We were able to translate the spectroscopic properties of the innovative bipartite ligands into the formation of highly luminescent europium complexes that exhibit efficient emission (ϕ(se)>0.1) upon sensitization in the near-visible region, that is, with an excitation wavelength above 350 nm. The luminescence of the Eu(3+) complexes is clearly detectable at concentrations as low as 10 pM. Furthermore, the structural organization of these bipartite ligands makes the complexes highly soluble in aqueous solutions and chemically stable over time.

Prevalence of urologic sequelae and bladder and bowel dysfunctions in patients with congenital Zika syndrome: A multicenter evaluation of the Zika virus bladder and bowel sequelae assistance network.

INTRODUCTION: Neurogenic bladder was first confirmed as a urological sequela of Congenital Zika Syndrome (CZS) in 2018. Further clinical-epidemiological evidence also confirmed neurogenic bowel dysfunction and cryptorchidism. To strengthen the care for these children, the Congenital Zika Virus Bladder and Bowel Sequelae Network (RASZ in Brazilian) was created, including six integrated centers in Brazil. This article represents the initial outcome of the efforts by RASZ. OBJECTIVE: To evaluate the prevalence of bladder and bowel dysfunction, cryptorchidism and other urological sequelae related to CZS in cohorts attended in six Brazilian states. STUDY DESIGN: Observational, prospective, multicenter study including children with CZS assisted in one of six RASZ collaborative centers between June 2016 and February 2023. Data were collected from patient's first assessment using the same protocols for urological and bowel evaluation. Categorical variables were analyzed by frequency of occurrence and numerical variables by mean, median, and standard deviation. The study was approved by the Research Ethics Committees of each center, all parents/caregivers provided written informed consent. RESULTS: The study included 414 children aged 2 months to 7 years (mean 2.77 years, SD 1.73), 227 (54.8 %) were male and 140 (33,8 %) referred urological and bowel symptoms on arrival. Prevalence of both urological and bowel sequelae was 66.7 %, 51 % of children aged 4 years and older had urinary incontinence (UI). UTI was confirmed in 23.4 % (two presented toxemia) and among males, 18.1 % had cryptorchidism. Renal ultrasonography, performed in 186 children, was abnormal in 25 (13.4 %), 7 had hydronephrosis. Among the 287 children who performed urodynamics, 283 (98.6 %) were altered: 232 had a lower bladder capacity, 144 a maximum bladder pressure of ≥40 cm H2O, and 127 did not satisfactorily empty their bladder. DISCUSSION: A higher prevalence of NLUTD, neurogenic bowel and cryptorchidism was confirmed in children with CZS. Early diagnosis and appropriate treatment, including a multidisciplinary approach, may reduce the risk of UTIs, UI and kidney damage. A limitation of the study was the inability of children to complete the protocol, specifically urodynamic evaluation, and ultrasonography. In both exams, the percentage of abnormal cases was higher than that expected in the normal population. CONCLUSION: A 66,7 % prevalence of combined urological sequelae and bladder-bowel dysfunction related to CZS was confirmed in patients evaluated in six Brazilian cohorts. The most frequent changes were related to NLUTD, neurogenic bowel, and cryptorchidism. Prevalence may be underestimated due to access restrictions to diagnostic tests.

Efficient greenish blue electrochemiluminescence from fluorene and spirobifluorene derivatives.

The spectroscopic and electrochemical behavior as well as electrogenerated chemiluminescence (ECL) of a series of donor-π-donor derivatives bearing triphenylamine groups as donor connected to a fluorene, 2,7-bis-(4-(N,N-diphenylamino)phen-1-yl)-9,9'-dimethylfluorene (1), or spirobifluorene core, 2,7-bis-(4-(N,N-diphenylamino)phen-1-yl)-9,9'-spirobifluorene (2) and 2,2',7,7'-tetrakis(4-(N,N-diphenylamino)phen-1-yl)-9,9'-spirobifluorene (3), were investigated. Besides a high photoluminescence (PL) quantum yield in solution (between 81 and 87%), an efficient radical ions annihilation process induces intense greenish blue ECL emission that could be seen with the naked eye. Only the tetrasubstituted spirobifluorene derivative (compound 3) shows weak ECL obtained by a direct annihilation mechanism. Because the energy of the annihilation reaction is higher than the energy required to form the singlet excited state, the S-route could be considered the pathway followed by the ECL process in these molecules. The ECL emissions recorded by direct ion-ion annihilation show two bands compared to the single structureless PL band. The ECL spectra obtained by a coreactant approach using benzoylperoxide as a coreagent show no differences relative to that produced by annihilation, except for an increasing of ECL intensity for all compounds.

Recent Advances in Injectable Hydrogels for Controlled and Local Drug Delivery.

Injectable hydrogels have received considerable interest in the biomedical field due to their potential applications in minimally invasive local drug delivery, more precise implantation, and site-specific drug delivery into poorly reachable tissue sites and into interface tissues, where wound healing takes a long time. Injectable hydrogels, such as in situ forming and/or shear-thinning hydrogels, can be generated using chemically and/or physically crosslinked hydrogels. Yet, for controlled and local drug delivery applications, the ideal injectable hydrogel should be able to provide controlled and sustained release of drug molecules to the target site when needed and should limit nonspecific drug molecule distribution in healthy tissues. Thus, such hydrogels should sense the environmental changes that arise in disease states and be able to release the optimal amount of drug over the necessary time period to the target region. To address this, researchers have designed stimuli-responsive injectable hydrogels. Stimuli-responsive hydrogels change their shape or volume when they sense environmental stimuli, e.g., pH, temperature, light, electrical signals, or enzymatic changes, and deliver an optimal concentration of drugs to the target site without affecting healthy tissues.

Use of plant extracts and essential oils in the control of bovine mastitis.

Bovine mastitis is the most important disease affecting dairy herds worldwide, causing direct impacts on farms' profitability and food safety issues. The prevention and treatment of this pathology is especially done through antimicrobials, but the increasing antimicrobial resistance of pathogens to this disease may affect the efficiency of conventional drugs. Besides, antimicrobials residues in milk and the environment are a potential threat to human health. Thereby, the use of plant extracts and essential oils may become promising alternatives for the control of bovine mastitis. Antimicrobial properties present in several plants are well described and plant extracts and essential oils are often considered safe to animals, humans and environment. This review summarizes the current problems encountered in the conventional treatment of mastitis, the possibilities of the use of plant extracts and essential oils as alternative agents for the control of these pathogens and the limitations found in the use of these plant derivatives. Finally, the perspectives to the use of plant extracts and essential oils for the treatment of bovine mastitis are presented.

Deformable Polyurethane Joints and Fibre Grids for Resilient Seismic Performance of Reinforced Concrete Frames with Orthoblock Brick Infills.

The behaviour of reinforced concrete frames with masonry wall infills is influenced a lot by the stiffness and strength difference between the frame and the infill, causing early detrimental damage to the infill or to the critical concrete columns. The paper reports the results from shake table seismic tests on a full-scale reinforced concrete (RC) frame building with modified hollow clay block (orthoblock brick) infill walls, within INMASPOL SERA Horizon 2020 project. The building received innovative resilient protection using Polyurethane Flexible Joints (PUFJs) made of polyurethane resin (PU), applied at the frame-infill interface in different schemes. Further, PUs were used for bonding of glass fibre grids to the weak masonry substrate to form Fibre Reinforced Polyurethanes (FRPUs) as an emergency repair intervention. The test results showed enhancement in the in-plane and out-of-plane infill performance under seismic excitations. The results confirmed remarkable delay of significant infill damages at very high RC frame inter-story drifts as a consequence of the use of PUFJs. Further, the PUFJ protection enabled the resilient repair of the infill even after very high inter-story drift of the structure up to 3.7%. The applied glass FRPU system efficiently protected the damaged infills against collapse under out-of-plane excitation while they restored large part of their in-plane stiffness.

Self-assembled multilayer surfaces of highly fluorescent spirobifluorene-based dye for label-free protein recognition.

The preparation of smart surfaces for protein detection is a challenging field of research. With the aim to achieve label-free detection in the solid state, we report on the organic surface functionalization for protein recognition without the need of previous chemical modification of the fluorophore. Layer-by-layer deposition of polyelectrolyte poly(vinyl benzyl tetramethylammonium) chloride (p(VBTMA)Cl) and a tetrasulfonate water-soluble low molecular weight fluorophore (1) based on spirobifluorene leads to modified glass and quartz substrates with outstanding photophysical properties in response to bovine serum albumin (BSA). The absorbance, photoluminescence as well as the fluorescence lifetimes were recorded for all surfaces. The surface structure and height of the different number of bilayers polymer/fluorophore were characterized by atomic force microscopy and ellipsometry. The results show linear trends in the absorption, fluorescence and height of the multilayer with increasing number of functionalization steps. Upon incubation with BSA the multilayer shows an increase in fluorescence up to 3-fold, which is also detectable with the naked eye. In conclusion, we report an easy, fast and biocompatible approach for the construction of protein sensors by self-assembly.

A highly fluorescent water soluble spirobifluorene dye with a large Stokes shift: synthesis, characterization and bio-applications.

The first water-soluble spirobifluorene derivative has been synthesized, which exhibits high fluorescence quantum yield and a large Stokes shift (>100 nm). Proteins induce changes in the emission color, allowing to reach the nanomolar detection limit. Cellular uptake and cytotoxicity studies in living cells revealed its biocompatibility, indicating potential application for live cell imaging.

Solid state electrochemiluminescence from homogeneous and patterned monolayers of bifunctional spirobifluorene.

Electrochemiluminescence (ECL) generated by a monolayer of a spirobifluorene derivative covalently bound onto an indium tin oxide (ITO) substrate is reported for the first time. Our approach allows the efficient preparation homogeneous and patterned substrates through micromolding in capillaries (MIMIC), and opens novel scenarios for multicolour ECL applications.

Author Correction: The Staphylococcus aureus extracellular matrix protein (Emp) has a fibrous structure and binds to different extracellular matrices.

A correction to this article has been published and is linked from the HTML and PDF versions of this paper. The error has been fixed in the paper.