Hyperpigmentation induced by topical 5-aminolaevulinic acid plus visible light

Photodynamic therapy (PDT) with 5-aminolaevulinic acid (ALA) is an alternative tool for the treatment of superficial non-melanoma skin cancers. Recently ALA-PDT has been employed with encouraging results also for warts, condylomata and psoriasis. In this study the effects of topical ALA plus irradiation with visible light on intact human skin have been evaluated. Five skin areas (A, B, C, D, and E) on the inner upper part of the arms of five healthy volunteers (skin types III and IV) were treated with (A) ALA 20% in base cream without irradiation, (B) only the vehicle (base cream) without ALA, (C, D and E) ALA cream at the concentrations of 5, 10 and 20%, respectively; all treatments were applied with an occlusive dressing. Four hours after ALA or vehicle application areas B, C, D and E were irradiated with a fixed dose of 40 J/cm(2). ALA penetration through the intact skin was evaluated by in vivo fluorescence determination. The effects on healthy skin were evaluated by clinical and chromometric examinations, light microscopy, immunohistochemistry and electron microscopy. RESULTS: (1) in vivo fluorescence demonstrated that ALA is able to penetrate through the intact skin, when applied with occlusive dressing and induces a classical fluorescence peak due to Protoporphyrin IX (PpIX) formation, which is the active photosensitiser. (2) Skin areas receiving ALA plus irradiation showed erythema and swelling just after the irradiative session and hyperpigmentation 48-72 h later. (3) Colourimetric data confirmed significant skin colour changes: values a* (representing the erythematous changes) increased only on the skin areas where ALA+irradiation were applied and during the 48 h after irradiation, thereafter a* began to decrease; values L* (pigmentation) increased during the 2 weeks following treatment. (4) Histopathological, immunohistochemical (S100, HMB-45) and electron microscopic findings showed an absolute increment of the number of melanocytes, which appeared clearly activated. In conclusion the application of ALA cream followed by irradiation is able to induce a pigmentation response in healthy human skin, at least in skin types III and IV. This melanocytic activation could have a potential for the treatment of skin disorders characterised by hypopigmentation.     

Capillary electrophoresis–electrospray mass spectrometry and HR–ICP–MS for the detection and quantification of 10B-boronophenylalanine (10B–BPA) used in boron neutron capture therapy

Boron neutron capture therapy (BNCT) is a bimodal radiotherapeutic treatment based on the irradiation of neoplastic tissues with neutrons after the tissues have selectively accumulated molecules loaded with nuclides with large neutron capture cross-sections (such boron-10). Boron-10 carriers have been tested to a limited extent, and clinical trials have been conducted on sulfhydryl borane (¹⁰B-BSH) and boronophenylalanine (¹⁰B-BPA). However, precise and accurate measurements of boron-10 concentrations (0.1–100 μg/g) in specimens and samples of limited size (μg scale) are needed in order to be able to biologically characterise new compounds in predictive tissue dosimetry, toxicology and pharmacology studies as well as in clinical investigations. A new approach based on fast separation and detection of ¹⁰B-BPA performed by coupling capillary electrophoresis to electrospray mass spectrometry is reported. This method allows the quantitative analysis and characterisation of ¹⁰B-BPA in a short time with a high separation efficiency. Detection limits of 3 μM for ¹⁰B-BPA and 30 ng/mL for ¹⁰B were obtained with CE–ESI–MS. A quantification limit of 10 μM for ¹⁰B-BPA (100 ng/mL for ¹⁰B) was attained. The total boron-10 concentration was determined by high-resolution inductively coupled mass spectrometry in order to validate the method. Boron-10 isotope measurements were carried out by HR–ICP–MS at medium resolution (R=4000) due to the presence of an isobaric interference at mass 10. Good agreement was obtained between the values from CE–ESI–MS and those from HR–ICP–MS. The method has been successfully used to determine the ¹⁰B-BPA in two lines of cultured cells.     

Fast atom bombardment mass spectrometry of reaction products between C3O2 and stabilized phosphorus ylides

A fast atom bombardment (FAB) mass spectrometric study on the open-chain compound 1,3-bis(cyanomethylenetriphenylphosphorane)propane-1,3-dione and on the cyclic zwitterionic compounds 4-oxy-5-triphenylphosphonium-6-methyl-2-pyrone and 4-oxy-5-triphenylphosphonium-6-phenyl-2-pyrone, obtained by reaction of carbon suboxide, C₃O₂, with stabilized phosphorus ylides, Ph₃PCHX (X?CN, COMe, COPh), is described. The FAB mass spectrometric behaviour of these compounds is compared with that shown by tri-phenylphosphoranilideneketene, Ph₃P ? C ? C ? O, and by 4-hydroxy-6-methyl-2-pyrone, with the aid of metastable ion data and collision spectroscopy.     

Effect of head group size, temperature and counterion specificity on cationic micelles

The critical micelle concentration (cmc) and ionisation degree (α), of micelles of cetyltrimethylammonium bromide (CTABr), cetyltrimethylammonium chloride (CTACl), cetyltripropylammonium bromide (CTPABr) and cetyltripropylammonium chloride (CTPACl) have been measured over a narrow temperature range at 2 degree intervals using electrical conductivity. CTPACl and CTPABr are very soluble in water and were measured in the temperature range 275.15-323.15K. The Krafft temperatures for CTABr and for CTACl are 293.15K and 284.15K, respectively and established a lower temperature limit for our studies on these two surfactants. The cmc vs temperature curves have a smooth minimum near room temperature and α linearly increases with temperature. The changes of cmc and α with temperature are smaller than those associated with the modification of head group size or counterion nature. Using these results, basic thermodynamic quantities associated with the phenomena of micellization have been evaluated. Thermodynamic properties of the surfactant solutions were discussed in terms of temperature dependence of the free energy, enthalpy and entropy of micellization. A close similarity between the effects of change in temperature on protein folding and micellization process appears from the data.     

Using multidimensional projection techniques for reaching a high distinguishing ability in biosensing

Recent advances in the control of molecular engineering architectures have allowed unprecedented ability of molecular recognition in biosensing, with a promising impact for clinical diagnosis and environment control. The availability of large amounts of data from electrical, optical, or electrochemical measurements requires, however, sophisticated data treatment in order to optimize sensing performance. In this study, we show how an information visualization system based on projections, referred to as Projection Explorer (PEx), can be used to achieve high performance for biosensors made with nanostructured films containing immobilized antigens. As a proof of concept, various visualizations were obtained with impedance spectroscopy data from an array of sensors whose electrical response could be specific toward a given antibody (analyte) owing to molecular recognition processes. In addition to discussing the distinct methods for projection and normalization of the data, we demonstrate that an excellent distinction can be made between real samples tested positive for Chagas disease and Leishmaniasis, which could not be achieved with conventional statistical methods. Such high performance probably arose from the possibility of treating the data in the whole frequency range. Through a systematic analysis, it was inferred that Sammon's mapping with standardization to normalize the data gives the best results, where distinction could be made of blood serum samples containing 10(-7) mg/mL of the antibody. The method inherent in PEx and the procedures for analyzing the impedance data are entirely generic and can be extended to optimize any type of sensor or biosensor.     

Sieve-based device for MALDI sample preparation. III. Its power for quantitative measurements

The solid sample inhomogeneity is a weak point of traditional MALDI deposition techniques that reflects negatively on quantitative analysis. The recently developed sieve-based device (SBD) sample deposition method, based on the electrospraying of matrix/analyte solutions through a grounded sieve, allows the homogeneous deposition of microcrystals with dimensions smaller than that of the laser spot. In each microcrystal the matrix/analyte molar ratio can be considered constant. Then, by irradiating different portions of the microcrystal distribution an identical response is obtained. This result suggests the employment of SBD in the development of quantitative procedures. For this aim, mixtures of different proteins of known molarity were analyzed, showing a good relationship between molarity and intensity ratios. This behaviour was also observed in the case of proteins with quite different ionic yields. The power of the developed method for quantitative evaluation was also tested by the measurement of the abundance of IGPP[Oxi]GPP[Oxi]GLMGPP (m/z 1219) present in the collagen-α-5(IV) chain precursor, differently expressed in urines from healthy subjects and diabetic-nephropathic patients, confirming its overexpression in the presence of nephropathy. The data obtained indicate that SBD is a particularly effective method for quantitative analysis also in biological fluids of interest.     

Two step, hydrolytic-solvothermal synthesis of redispersible titania nanocrystals and their gas-sensing properties

A titanium chloromethoxide solution was prepared by reacting TiCl₄ with methanol, followed by water addition. The starting solutions were characterized by Fourier Transform Infrared (FTIR) spectroscopy, evidencing that the in situ generated water results in early hydrolysis of the chloroalkoxide. The solution was reacted with molten dodecylamine at room temperature, obtaining a white slurry of amorphous titania nanoparticles. Stable, redispersible TiO₂ nanocrystals could be prepared by subsequent solvothermal treatment in oleic acid at 250???C. The use of oleic acid was essential for obtaining crystalline structures, while other surfactants prevented crystallization. The nanocrystals were characterized by X-ray Diffraction and Transmission Electron Microscopy, confirming the formation of anatase TiO₂ nanocrystals with a mean size of 3.3?nm. The TiO₂ nanocrystals were used for fabricating gas-sensing devices, which were tested towards ethanol vapors. The initial small size of the nanocrystals, and the limited size growth during the high-temperature sensor operation, result in remarkable sensing performances if compared with bulk titania sensors.     

Synthesis,characterization and antitumor activity of palladium(II) complexes of monoethyl 8-quinolylmethylphosphonate

The complex-forming properties of monoethyl 8-quinolylmethylphosphonate (8-Hmqmp) towards palladium(II) ion have been investigated by reactions of the hydrochloride, 8-Hmqmp · HCl · H₂O, and sodium salt, Na(8-mqmp) · 2H₂O, of this monoester with palladium(II) halide compounds in aqueous solution over a wide pH range. Depending on pH and initial quinolinium and palladium salts, four types of complexes have been formed. Under acidic solution the ion-pair salt complexes [8-H₂mqmp]₂[PdX₄] (1 and 2, pH < 3) and [8-H₂mqmp]₂[Pd₂X₆] (3 and 4, pH ∼ 3), with protonated quinoline ligand as cation and tetrahalopalladate or hexahalodipalladate complex as anion (X = Cl, Br), were isolated. By heating in methanol the chloro complexes 1 and 3 as well as bromo complexes 2 and 4 were converted into the quinolinium salt complexes, [8-H₂mqmp][Pd(8-Hmqmp)X₃], 5 and 6, respectively, containing as anion the quinoliniummethylphosphonatetrihalopalladate complex with palladium bonded at the phosphonic acid moiety. The chelate complex 7, [Pd(8-mqmp)₂], with ligand bonded through the quinoline nitrogen and the deprotonated phosphonic acid oxygen and forming two seven-membered {N,O} chelate rings, was obtained in neutral and basic media. The complexes were identified and characterized by elemental analysis, magnetic and conductance measurements, spectroscopic studies (IR, ¹H NMR, UV–Vis, positive/negative ion FAB MS) and thermal analysis (TG, DTA). As a preliminary screening for their biological activity, complexes were investigated for their ability to inhibit the cancer growth in vitro in the human KB and murine L1210 cell lines. The results obtained were compared with those obtained for the complexes of diethyl 8-quinolylmethylphosphonate (8-dqmp) and monoethyl 2-quinolylmethylphosphonate (2-Hmqmp), and structural factors that determine the complex activity were discussed.