Transformation of COUPY fluorophores into a novel class of visible light-cleavable photolabile protecting groups

Although photolabile protecting groups (PPGs) have found widespread applications in several fields of chemistry, biology and materials science, there is a growing interest in expanding the photochemical toolbox to overcome some of the limitations of classical caging groups. In this work, the synthesis of a new class of visible-light-sensitive PPGs based on low-molecular weight COUPY fluorophores with several attractive properties, including long-wavelength absorption, is reported. Besides being stable to spontaneous hydrolysis in the dark, COUPY-based PPGs can be efficiently photoactivated with yellow (560 nm) and red light (620 nm) under physicological-like conditions, thereby offering the possibility of unmasking functional groups from COUPY photocages under irradiation conditions in which other PPGs remain stable. Additionally, COUPY photocages exhibit excellent cellular uptake and accumulate selectively in mitochondria, opening the door to delivering caged analogues of biologically active compounds into this organell

Mitochondria-Targeted COUPY Photocages: Synthesis and Visible-Light Photoactivation in Living Cells

Releasing bioactive molecules in specific subcellular locations from the corresponding caged precursors offers great potential in photopharmacology, especially when using biologically compatible visible light. By taking advantage of the intrinsic preference of COUPY coumarins for mitochondria and their long wavelength absorption in the visible region, we have synthesized and fully characterized a series of COUPY-caged model compounds to investigate how the structure of the coumarin caging group affects the rate and efficiency of the photolysis process. Uncaging studies using yellow (560 nm) and red light (620 nm) in phosphate-buffered saline medium have demonstrated that the incorporation of a methyl group in a position adjacent to the photocleavable bond is particularly important to fine-tune the photochemical properties of the caging group. Additionally, the use of a COUPY-caged version of the protonophore 2,4-dinitrophenol allowed us to confirm by confocal microscopy that photoactivation can occur within mitochondria of living HeLa cells upon irradiation with low doses of yellow light. The new photolabile protecting groups presented here complement the photochemical toolbox in therapeutic applications since they will facilitate the delivery of photocages of biologically active compounds into mitochondria

Thermoresponsive zwitterionic poly(phosphobetaine) microgels: Effect of macro-RAFT chain length and cross-linker molecular weight on their antifouling properties

Adsorption of proteins on biological surfaces is a detrimental phenomenon that reduces the work-life of the implants in various biomedical applications. Here, we synthesized a new class of thermoresponsive zwitterionic poly(phosphobetaine) (PMPC) microgel with excellent surface antifouling property by macro-RAFT mediated thiol-epoxy click reaction. End-group modified zwitterionic PMPC homopolymers with well-defined molecular weight and narrow dispersity were grafted onto poly(N-vinylcaprolactam-co-glycidyl methacrylate) (PVG) copolymer backbone followed by addition of a cross-linker, leading to microgel formation. While no upper critical solution temperature (UCST) was found in poly(N-vinylcaprolactam-co-glycidyl methacrylate-g-2-methacryloyloxyethyl phosphorylcholine) (PVGP) graft copolymers, the corresponding microgels exhibited both UCST and lower critical solution temperature (LCST) transitions, related to the swelling and collapse of PMPC and poly (N-vinylcaprolactam) (PVCL) components respectively. An increase in the molecular chain length of the PMPC increased the shifting of UCST and LCST of the microgels to higher temperatures, due to the ability of zwitterionic groups to coordinate a large number of water molecules. The effect of the variation in the molecular weights of amphiphilic poly(ethylene glycol) diamine (PEG-NH2) cross-linker was also reflected in both temperature and salt responsiveness of the microgels. The efficacy of the microgels as potential antifouling materials was further studied by fluorescence microscopy and XPS analysis on microgel coatings treated with FITC-BSA solution and pure BSA solution respectively. Lower protein adsorption was observed for microgels grafted with higher molecular chain length of PMPC, whereas, the microgels synthesized using higher molecular weight PEG-NH2 diamine cross-linker displayed greater protein adsorption on their surfaces

Efektivitas Ekstrak Batang Arcangelisia flava Merr. sebagai Afrodisiaka pada Mencit Jantan dan Betina

This study aimed to analyze the effect of the stem extract of the yellow root plant (Arcangelisia flava Merr.) from Enggano Island on the introduction and locomotor activity of male and female mice (Mus musculus L.). The method used is an experiment with a completely randomized design. The treatment variations were extracted at a dose of 150 mg/kgBW, 200 mg/kgBW and 250 mg/kgBW, Neo Hormoviton capsules at a 200 mg/kgBW dose as a positive control, and without extract or strong drug as a negative control. The results showed that the administration of the extract caused an increase in the introduction activity, but the activity decreased with the increase in the dose of the given extract. Meanwhile, an increase in locomotor activity occurred at a dose of 250 mg/kgBW, while the doses of 150 mg/kgBW and 200 mg/kgBW tended to cause a decrease. Duncan's further test showed that a dose of 250 mg/kgBW increased locomotor activity which was more significant in female mice but not significantly in male mice. In conclusion, the extract at a dose of 250 mg/kgBW can increase locomotor activity, higher than Neo Hormoviton capsules at 200 mg/kgBW in both male and female mice. Keywords: Aphrodisiac, Sexual Activity, Arcangelisia flava, Fitnes

High photostability in non-conventional coumarins with far-red/NIR emission through azetidinyl substitution

Replacement of electron-donating N,N-dialkyl groups with three or four-membered cyclic amines (e.g., aziridine and azetidine, respectively) has been described as a promising approach to improve some of the drawbacks of conventional fluorophores, including low fluorescent quantum yields (F) in polar solvents. In this work we have explored the influence of azetidinyl substitution on non-conventional coumarin-based COUPY dyes. Two azetidine-containing scaffolds were first synthesized in four linear synthetic steps and easily transformed into far-red/NIR-emitting fluorophores through N-alkylation of the pyridine moiety. Azetidine introduction in COUPY dyes resulted in enlarged Stokes' shifts with respect the N,N-dialkylamino-containing parent dyes, but the F were not significantly modified in aqueous media, which is in contrast with previously reported observations in other fluorophores. However, azetidinyl substitution led to an unprecedented improvement in the photostability of COUPY dyes and high cell permeability was retained since the fluorophores accumulated selectively in mitochondria and nucleoli of HeLa cells. Overall, our results provide valuable insights for the design and optimization of novel fluorophores operating in the far-red/NIR region, since we have demonstrated that three important parameters (Stokes' shifts, F and photostability) cannot be always simultaneously addressed by simply replacing a N,N-dialkylamino group with azetidine, at least in non-conventional coumarin-based fluorophores

Assessing amphiphilic ABAB Zn(II) phthalocyanines with enhanced photosensitization abilities in in vitro photodynamic therapy studies against cancer

We have previously demonstrated that singlet oxygen photosensitization abilities of Zn(II) phthalocyanines (Zn(II)Pcs) are enhanced through α-functionalization with bulky fluorinated substituents (i.e., bis(trifluoromethyl)phenyl units) at facing positions of ABAB Zn(II)Pcs, where A and B refer to differently functionalized isoindoles. In this work, we have prepared the Zn(II)Pc ABAB 1 endowed with hydrophilic triethylene glycol monomethyl ether (i.e., at the A isoindoles) to provide solubility in aqueous media, together with its A3B and A4 counterparts, and compared their ability to behave as photosensitizers for photodynamic therapy. All photophysical data, aggregation studies and preliminary in vitro biological assays in cell cultures of SCC-13 (squamous cell carcinoma) and HeLa (cervical cancer cells), have proved ABAB 1 as the best photosensitizer of the seriesThis research was funded by MINECO, Spain (CTQ2017-85393-P and CTQ2016-78454-C2-1-R) , Instituto de Salud Carlos III and Feder Funds (FIS PI18/00708) and ERANET/MINECO EuroNanoMe

Filamentous sieve element proteins are able to limit phloem mass flow, but not phytoplasma spread

In Fabaceae, dispersion of forisomes\u2014highly ordered aggregates of sieve element proteins\u2014in response to phytoplasma infection was proposed to limit phloem mass flow and, hence, prevent pathogen spread. In this study, the involvement of filamentous sieve element proteins in the containment of phytoplasmas was investigated in non-Fabaceae plants. Healthy and infected Arabidopsis plants lacking one or two genes related to sieve element filament formation\u2014AtSEOR1 (At3g01680), AtSEOR2 (At3g01670), and AtPP2-A1 (At4g19840)\u2014were analysed. TEM images revealed that phytoplasma infection induces phloem protein filament formation in both the wild-type and mutant lines. This result suggests that, in contrast to previous hypotheses, sieve element filaments can be produced independently of AtSEOR1 and AtSEOR2 genes. Filament presence was accompanied by a compensatory overexpression of sieve element protein genes in infected mutant lines in comparison with wild-type lines. No correlation was found between phloem mass flow limitation and phytoplasma titre, which suggests that sieve element proteins are involved in defence mechanisms other than mechanical limitation of the pathogen

Tongue stretching: technique and clinical proposal.

Abstract Objectives The tongue is an organ with multiple functions, from sucking to phonation, from swallowing to postural control and equilibrium. An incorrect position or mechanics of the tongue can causes sucking problems in the newborn or atypical swallowing in the adult, with repercussions on the position of the head and neck, up to influencing upright posture and other problems. Tongue dysfunctions are quite frequent (10–15%) in the population. For the manual therapist, this frequency indicates one to two subjects every 30 patients. Exercises have been proposed to improve the tone and strength of the swallowing muscles but the results are not so clear in the literature. The aim of this study is to describe and provide a tongue muscle normalization technique that helps the manual therapist in the treatment of problems related to it. Methods The literature has been investigated through pubmed, Google scholar of the last 10 years, the keywords used and combined with the Boolean operators AND and OR, are: "tongue, tongue habits, tongue diseases, taste disorder, neck pain, posture, postural balance, atypical swallowing, muscle stretching exercise, tissue expansion, soft tissue therapy, osteopathic manipulative treatment". Results and Conclusions The technique is possible to be executed even in a sitting position, in the case the patient is unable to assume a supine position, the subject should provides immediate feedback that allows the therapist to understand if the technique has been correctly executed. The simplicity of execution and application of the technique makes it a possible and immediate therapeutic tool in the clinical setting

Redesigning the coumarin scaffold into small bright fluorophores with far-red to NIR emission and large Stokes' shifts useful for cell imaging

Among the palette of previously described fluorescent organic molecules, coumarins are ideal candidates for developing cellular and molecular imaging tools due to their high cell permeability and minimal perturbation of living systems. However, blue-to-cyan fluorescence emission is usually difficultin in vivo applications due to the inherent toxicity and poor tissue penetration of short visible light wavelengths. Here, we introduce a new family of coumarin-based fluorophores, nicknamed COUPY, with promising photophysical properties, including emission in the far-red/near-infrared (NIR) region, large Stokes shifts, high photostability, and excellent brightness. COUPY fluorophores were efficiently synthesized in only three linear synthetic steps from commercially available precursors, with the N-alkylation of a pyridine moiety being the key step at the end of the synthetic route, as it allows for the tuning of the photophysical properties of the resulting dye. Owing to their low molecular weights, COUPY dyes show excellent cell permeability and accumulate selectively in nucleoli and/or mitochondria of HeLa cells, as their far-red/NIR fluorescence emission is easily detected at a concentration as low as 0.5 μ M after an incubation of only 20 min. We anticipate that these coumarin scaffolds will open a way to the development of novel coumarin-based far-red to NIR emitting fluorophores with potential applications for organelle imaging and biomolecule labeling

Cellular distribution of the histamine H3 receptor in the basal ganglia : functional modulation of dopamine and glutamate neurotransmission

This is the author's version of a work that was accepted for publication in Basal ganglia. Changes resulting from the publishing process, such as peer review, editing, corrections, structural formatting, and other quality control mechanisms may not be reflected in this document. Changes may have been made to this work since it was submitted for publication. A definitive version was subsequently published in Vol. 3 Núm. 2 (Jul. 2013)Altres ajuts: Red_de_Trastornos_Adictivos/RD06/0001/0015Histamine H3 receptors (H3R) are widely expressed in the brain where they participate in sleep-wake cycle and cognition among other functions. Despite their high expression in some regions of the basal ganglia, their functional role in this forebrain neural network remains unclear. The present findings provide in situ hybridization and immunohistochemical evidence for H3R expression in several neuronal populations of the rat basal ganglia but not in astrocytes (glial fibrillary acidic protein immunoreactive cells). We demonstrate the presence of H3R mRNA and protein in dopaminergic neurons (tyrosine hydroxylase positive) of the ventral tegmental area and substantia nigra. In the dorsal and ventral (nucleus accumbens) striatal complex we show H3R immunoreactivity in cholinergic (choline acetyltransferase immunoreactive) and GABAergic neurons (substance P, proenkephalin or dopamine D1 receptor positive) as well as in corticostriatal terminals (VGLUT1-immunoreactive). Double-labelling experiments in the medial prefrontal cortex show that H3R is expressed in D1R-positive interneurons and VGLUT1-positive corticostriatal output neurons. Our functional experiments confirm that H3R ligands modulate dopamine synthesis and the probability of glutamate release in the striatum from cortico-striatal afferents. The presence of H3R in such different neuronal populations and its involvement in the control of striatal dopaminergic and glutamatergic transmission ascribes a complex role to H3R in the function of the basal ganglia neural network