Dependence of fibroblast autofluorescence properties on normal and transformed conditions. Role of the metabolic activity

The dependence of autofluorescence properties on the metabolic and functional engagement and on the transformation condition was studied on single cells. Normal Galliera rat fibroblasts at low subculture passage (cell strain), at high subculture passage (stabilized cell line), and transformed cell line derived from a rat sarcoma were used as a cell model. The study was performed by microspectrofluorometric and fluorescence imaging technique. The autofluorescence properties of cells were studied by excitation at two wavelengths, namely 366 nm and 436 nm, that are known to favor the emission of different fluorophores. Spectral shape analysis indicated that under excitation at 366 nm autofluorescence is ascribable mainly to coenzyme molecules, particularly to reduced pyridine nucleotides, while under excitation at 436 nm, flavin and lipopigment emission is favored. The energetic metabolic engagement of the different cell lines was analyzed in terms both of parameters related to anaerobic-aerobic pathways (biochemical assay) and of mitochondrial features (supravital cytometry). The results showed that the cell strain and the stabilized and transformed cell lines can be distinguished from one another on the basis of both overall fluorescence intensity and the relative contributions of spectral components. These findings indicated a relationship between autofluorescence properties and energetic metabolism engagement of the cells that, in turn, is dependent on the proliferative activity and the transformed condition of the cells. In that it is a direct expression of the energetic metabolic engagement, autofluorescence can be assumed as an intrinsic parameter of the cell biological condition, suitable for diagnostic purposes.     

Enzyme-assisted Cell Photosensitization: A Proposal for an Efficient Approach to Tumor Therapy and Diagnosis. The Rose Bengal Fluorogenic Substrate

Abstract— Rose bengal, a xanthene derivative among the most efficient producer of singlet oxygen, was submitted to a chemical modification consisting in the introduction of an acetate group into the aromatic ring fluorophore structure. The acetate group acts as a quencher, thus inactivating both fluorescence and photosensitization properties of the molecule. In the modified structure, rose bengal acts as a fluorogenic substrate giving rise to the cellular reaction termed fluorochromasia. The acetate group is recognized by a carboxylic esterase activity that splits it. Removal of the quencher group results in restoring the native structure of photosensitizer inside the cells. The intracellular turnover of rose bengal acetate was studied in rat glioma-derived cultured cells, in terms of the balance of the processes of influx and enzyme hydrolysis of the fl0075 orogenic substrate, and of the efflux of the fluorescent product. A large intracellular accumulation of photosensitizer is obtained when treatments are performed with the fluorogenic substrate, even at the drug concentration at which rose bengal does not enter the cells. The intracellular localization allows rose bengal to exert a more effective photosensitization effect. Provided that the quencher group is selected according to the metabolic properties of the tumor cells, the use of fluorogenic substrates as photosensitizer precursors could improve fluorescence diagnosis and the photodynamic therapy of tumors, exploiting the biological properties that distinguish pathological from normal conditions.     

Non-steroidal antiinflammatory agents. VI. Synthesis of 10-oxo-5H-pyrrolo[1,2-b]isoquinoline-3-acetic Acid,a conformationally restricted analogue of tolmetin

The synthesis of 10-oxo-5H-pyrrolo[1,2-b]isoquinoline-3-acetic acid 4 , a tricyclic analogue of tolmetin which might show significant analgesic-antiinflammatory activity, has been accomplished in four steps. Alkylation of ethyl 2-pyrrolylglyoxylate ( 15 ) with 2-cyanobenzyl bromide afforded derivative 16 , which was transformed by the Huang-Minion reaction into the dicarboxylic acid 6 . Cyclization of this key intermediate to 17 , followed by alkaline hydrolysis, afforded 10-oxo-5H-pyrrolo[1,2–6]isoquinoline-3-acetic acid ( 4 ). Various attempted syntheses of 6 are also discussed.     

v-Triazolines—VI: Rearrangement of triazolines from 2-substituted-1-aryl-and 1-heteroaryl-1-amino-ethylenes and tosylazide

Reaction of tosylazide with 2-substituted, 1-aryl- or 1-heteroaryl-1-amino-ethylenes affords, via unstable triazolines, a zwitterionic intermediate which can lead (i) through nitrogen loss and rearrangement to amidine (2) and (ii) through C₄-C₅ cleavage to the formation of a diazo compound and amidine (3).Some aspects of the two mechanistic pathways are discussed.     

Electron spin resonance study of the radiation-induced polymerization of N-vinylcarbazole

When crystals of N-vinylcarbazole are γ-irradiated at 77°K., the ESR spectrum observed before warming consists of three peaks attributed to a radical–cation with the unpaired spin associated mainly with the nitrogen atom. Above 90°K. polymerization occurs, initiated by the cation, and the spectrum changes to that of an alkyl type of radical, ?N? ?H? CH₂, trapped in the polymer. Single crystals were used for a detailed analysis of the nuclear hyperfine parameters of the observed radicals.     

Determination of 4-nonylphenol and 4-nonylphenol ethoxylates in river sediments by microwave assisted solvent extraction

The efficiency of Microwave Assisted Solvent Extraction (MASE) for the simultaneous determination of nonylphenol (NP) and nonylphenol ethoxylates (NPE) in river sediments has been evaluated. An optimisation study was carried out in order to identify the variables having the greatest influence on the extraction efficiency. The comparison between the solvents (methanol and acetone-hexane 1:1) shows that the more polar solvent (i.e. methanol) allows a more effective extraction of NP and NPE from sediments. Analytical results show that there is not a considerable improvement by doubling extraction time, while the increase of solvent volume is significant. The comparative study with Soxhlet and pressurised liquid extraction (PLE) methods demonstrates that Microwave Assisted Solvent Extraction is a suitable alternative extraction method for the 4-NP and 4-NPE determination in river sediments, showing an accuracy and precision comparable to those obtained with PLE and better than those obtained with Soxhlet.     

Impacting the length of wormlike micelles using mixed surfactant systems

The effect of adding an alcohol ethoxylate nonionic surfactant (C(18)E(18)) to aqueous solutions of a cationic surfactant, erucyl bis(hydroxyethyl) methylammonium chloride (EHAC,CH(3)(CH(2))(7)(CH)(2)(CH(2))(12)N(+)-(CH(2)CH(2)OH)(2)CH(3)Cl(-)), was studied using small-angle neutron scattering (SANS), steady-state rheology, and cryo-transmission electron microscopy (Cryo-TEM). This cationic surfactant has the ability to self-assemble into giant wormlike micelles in the presence of an electrolyte, such as KCl. In salt-free solutions, the mixture of the two surfactants gave rise to spherical micelles. The scattering curves obtained were fitted with a polydisperse core-shell model combined with a Hayter Penfold potential. The inner and outer radii were found to be dependent on the surfactant ratio. In the presence of KCl, mixed wormlike micelles were formed. However, further addition of C(18)E(18) promoted the breaking of the micellar worms with the appearance of a structure peak in the scattering curves. In addition, it was found that the low shear viscosity is decreased upon addition of the alcohol ethoxylate nonionic surfactant. These findings are in good qualitative agreement with the Cryo-TEM images. The results show that the addition of the nonionic surfactant to the system is a method of controlling the worm length.     

Giant micellar worms under shear: a rheological study using SANS

Flow-SANS experiments were performed on viscoelastic aqueous solutions of erucyl bis(hydroxyethyl) methylammonium chloride in the presence of potassium chloride. This cationic surfactant has the ability to form very long and flexible wormlike micelles upon addition of salt. The effects of the key-parameters-shear rate, temperature, surfactant and salt concentration-on the ability of the micelles to align in the flow-field were investigated. The scattering data were analyzed in terms of an anisotropy factor (Af). It was found that the wormlike micelles aligned in the direction of the applied shear rate and that the anisotropy factor increased with shear rate. In addition, an increase in temperature caused a decrease of the anisotropy factor (Af) due to the formation of shorter worms. Furthermore, the branching of the micelles at high ionic strength caused the anisotropy factor to decrease in comparison with the values obtained from linear wormlike micelles, hence revealing that the formation of 3-way junctions restricts the alignment of the micelles in the shear-flow. Furthermore, the total surfactant concentration was found to affect the shear-induced patterns significantly, and different behaviors were observed depending on the ionic strength.     

Synthesis of arylsulfonyl substituted pyrroles

The cycloaddition reaction of symmetrically and unsymmetrically substituted munchnones with arylsulfonyl alkynes has been studied. The reaction affords pyrrole derivatives whose structures were assigned on the basis of spectroscopic data. The distribution of regioisomers observed in the case of unsymmetrically substituted munchnones is discussed.     

Sur l'evolution du rapport signal-sur-bruit dans la deconvolution quand le bruit est fonction du signal,en particulier pour des recepteurs “parfaits‘

The influence on the signal-to-noise ratio in deconvolutions carried out from recordings of a perfect detector is discussed with emphasis given to the following remark: the transformation of the mean square of the noise of elementary zones is linear as well as that of the signal itself.