No description available.

The objective of this study was to investigate the effects of melamine and ether contents on the curing and performance properties of UF resins as binders for wood composites. Various UF and UMF resins were synthesized with three different synthesis procedures. These resins were examined by 13C NMR, rheometer, and other methods and evaluated as particleboard binders. Three-layer particleboards were prepared with the resins catalyzed with various catalysts and levels, applied in face and core layers. The board test results were compared. Only about half of added melamine had reacted with formaldehyde. UMF resins were found to be catalyzed with stronger catalysts at suitable levels depending on melamine levels and on which layer of particleboard the UMF resins are to be applied. Even catalyzed with a stronger catalyst, the curing rates of UMF resins were still slower, and storage stabilities were shorter than UF resins, but the pot lives were longer, and internal bond strength and water resistance were higher. Moreover, resins synthesized with procedures 2 and 3 showed obviously longer storage times, longer pot lives, and longer gel times, and the particleboards bonded with these resins showed significant improvements in internal bond strength and water absorption values but the formaldehyde contents increased. The increased formaldehyde content test values indicated that linear methylene-ether groups in UF resins decompose in the hot-pressing of boards to emit formaldehyde, most of which is not captured back into the UF resin matrix. Uron-type methylene-ether groups decompose in the hot-pressing of boards to participate in the curing process and enhance the bonding of boards, but it could also emit extra formaldehyde which may not be effectively captured by UF resins but more effectively by UMF resins if the amount of melamine is high enough because of the increased reactive capacities of melamine. The results of this research offered a new hypothesis that the linear methyleneether bonds in UF resins might be a major contributor of the high free formaldehyde contents of particleboards. Decreasing the linear methylene-ether groups contents might effectively bring down the formaldehyde content of boards.

The response of loblolly pine (Pinus taeda L.) to fertilization and weed control at stand establishment, using various formulations of conventional and controlled release N fertilizers was evaluated in a greenhouse study and at two field trials in the Virginia Piedmont, in 2002 and 2003. The greenhouse study evaluated five fertilizer treatments; 1) check with no fertilizer; 2) granular ammonium nitrate (10-10-10 + micro-nutrients) applied to the soil surface; 3) granular methylene urea (40-0-0) applied to the soil surface; 4) methylene urea (20-10-5) in tablet form applied in the planting hole; and 5) isobutylidenediurea (IBDU) (9-9-4) in tablet form applied in the planting hole. Equal amounts of N and P were applied. Fertilization significantly increased seedling root collar diameter and volume at the end of the first growing season in the greenhouse study. Differences in diameter and volume were still significantly different late in the second growing season, however at the last measurement the differences were no longer significant. An analysis of transformed growth curves for 2003 indicated that the ammonium nitrate treated seedlings had a significantly steeper slope than all other treatments. In field trials, at an old-field site and a cut-over site, the same fertilizer products were tested, except granular diammonium phosphate (18-46-0) was used substituted for the ammonium nitrate. Higher rates of N and P were used in the field trials. Complete weed control increased seedling volume by over 700 % after two growing seasons at the reforested old-field site, however fertilizer effects were not significant. At the reforested cut-over site an interaction between weed control and fertilizer treatments was observed. The MU and DAP granular, and the IBDU tablet treatments each had significantly greater seedling volume than the check and the other tablet controlled release fertilizer. IBDU tablets appear to have high fertilizer efficiency, due to the slow release nature and are safe to place in close proximity to the seedling root system. / Master of Science

O aumento da tolerância aos fungicidas convencionais, observado em diversos gêneros de fungos de interesse, e a preocupação crescente com o meio ambiente têm estimulado o desenvolvimento de novas alternativas para o controle de fitopatógenos. Dentre as novas estratégias antimicrobianas que têm sido avaliadas, o tratamento fotodinâmico antimicrobiano (TFA) é uma das mais promissoras. O TFA baseia-se no uso de um fotossensibilizador (FS) que se acumula seletivamente nas células do microrganismo alvo. A ativação do FS pela luz, na presença de oxigênio molecular, induz à formação de espécies reativas de oxigênio, como o oxigênio singlete, que provocam a morte do microrganismo alvo, com poucos danos ao hospedeiro. Apesar de muito promissor, o TFA ainda não foi explorado para o controle de fitopatógenos. No presente estudo, foi avaliada, in planta, a eficácia do TFA com o fotossensibilizador azul de metileno (MB) e radiação solar em conídios do fungo fitopatogênico Colletotrichum acutatum. Os experimentos foram conduzidos com MB de diferentes fabricantes, em duas concentrações (25 e 50 ?M) e em diferentes formulações, ao longo das diferentes estações do ano, em dias com e sem nebulosidade e em diferentes temperaturas. O TFA com MB e radiação solar foi extremamente eficaz, matando aproximadamente 100% dos conídios em todas as condições avaliadas. Os efeitos da aplicação do MB na planta hospedeira Citrus sinensis também foram avaliados. O FS foi aplicado repetidas vezes em folhas e em flores de mudas de laranjeira. Nenhum dano foi observado nas folhas ou nas pétalas após os tratamentos com o MB. Pela primeira vez, foi feito o estudo mecanístico dos efeitos do TFA com MB em conídios de C. acutatum. Foram avaliados os efeitos do TFA na permeabilidade da membrana plasmática dos conídios e na formação de espécies reativas de oxigênio tardias, por meio da utilização das sondas fluorescentes iodeto de propídio (PI) e dihidroetídio (DHE), respectivamente. O TFA com MB danificou a membrana plasmática, reduzindo a sua permeabilidade seletiva e induziu a formação de espécies reativas de oxigênio tardias nos conídios. Também foi conduzido um estudo de longa duração, no qual foi avaliada a tolerância dos conídios de C. acutatum ao TFA com o novo azul de metileno, após cada um de 45 ciclos sucessivos de TFA, seguido pela seleção dos sobreviventes. Foram observados aumentos gradativos na tolerância ao TFA a partir do 21° ciclo de seleção. Além do aumento na tolerância, também foi observado um aumento da pigmentação dos conídios, provavelmente decorrente da seleção de mutantes que acumulam mais carotenóides. / The increasing tolerance to conventional fungicides observed in several genera of pathogenic fungi and the growing concern about the environment have stimulated the development of new alternatives to control of plant pathogens. Among the new antimicrobial strategies that have been evaluated, antimicrobial photodynamic treatment (APDT) is one of the most promising. APDT is based on the use of a photosensitizer (PS) that selectively accumulates in the target microbial cells. Activation of the PS by light in the presence of molecular oxygen induces the formation of reactive oxygen species, such as singlet oxygen, which cause damages to the target microorganism without significant harm to the host. Although very promising, APDT has not yet been used to control plant-pathogenic fungi. In the present study, the effectiveness of the APDT with the photosensitizer methylene blue (MB) and solar radiation on conidia of the plant-pathogenic fungus Colletotricum acutatum was evaluated in planta. The experiments were conducted with MB from different brands, in two concentrations (25 and 50 ?M) and different formulations, throughout the different seasons of the year, on cloudy and sunny days and at different temperatures. APDT with MB and solar radiation was extremely effective, killing around 100% of conidia in all conditions evaluated. The effects of the MB on the host plant Citrus sinensis were also evaluated. The PS was applied repeatedly on the leaves and flowers of the plants. No damage was observed on the leaves or the petals after treatment with MB. For the first time the mechanistic study evaluating the effects of APDT on C. acutatum conidia was carried out. The effects of APDT on the permeability of the conidial plasma membrane and on the production of secondary reactive oxygen species were evaluated by using the fluorescent probes propidium iodide (PI) and dihydroethidium (DHE), respectively. APDT with MB damaged the plasma membrane, reducing its selective permeability and induced the formation of secondary reactive oxygen species in conidia. It was also conducted a long term study which assessed the tolerance of conidia of C. acutatum to APDT, after each of 45 successive cycles of APDT with new methylene blue followed by the selection of the survivors. A gradual increase in APDT tolerance was observed from the 21st cycle. Besides the increase in tolerance to APDT, it was also observed an increase in the pigmentation of conidia probably due to the selection of mutants that accumulate more carotenoids.

Significant developments have been made in the field of C–H activation. However, various disadvantages, mainly low reactivity and selectivity, limit their usage in large-scale synthesis. It is crucial to understand the mechanisms and the nature of the transient species involved in the C–H activation paths to develop effective catalytic routes for homogeneous C–H functionalization reactions. Computational techniques are employed in this study to throw light on these processes. Chapter 1 briefly introduces C–H activation and functionalization reactions. After classifying the reactions on the basis of mechanisms, computational studies on the mechanisms of C–H activation reactions are described. The challenges involved in the discovery of efficient homogeneous C–H functionalization catalysts and progress made in the field are discussed. The insights provided to overcome the problems associated with the catalytic C–H functionalization reactions in a few examples are highlighted. In Chapter 2, DFT model studies are carried out to estimate the affinity and selectivity of 16-electron half-sandwich d6-metal fragments (η5–C5H5)Re(CO)2 and (η6–C6H6)W(CO)2 for binding with alkane C–H bonds. Different C–H binding sites of pentane, at the M06 level of theory have been evaluated. The effects of ancillary ligand variations on the metal–pentane binding strength are studied by substituting different ligands such as N-heterocyclic carbene (NHC), PF3 and NO+ for one of the carbonyl ligands. Isomers of the metal-pentane C–H σ-complexes studied in this chapter are shown in Scheme 1. Binding energies of the terminal methyl C–H bonds (C1 and C5) are significantly lower than those of the methylene C–H bonds (C2, C3 and C4) in all the cases. The metal–pentane binding interactions of the rhenium complexes are significantly stronger than those of the corresponding tungsten analogs. The PF3 complexes have slightly greater binding energies compared to the CO complexes, in both Re(I) and W(0) analogs. These results are in conformity with the experimental results. The electron-deficient nitrosyl complexes have the highest binding energies. These results illustrate that by proper tuning of the electronic factors of the transition-metal fragments with different ancillary ligands, the alkane C–H binding affinity can be controlled. Energy decomposition analyses (EDA) are carried out to determine the nature of the interaction between the metal fragments and pentane C–H bonds. Scheme 1. Formation of pentane C–H σ-complexes Chapter 3 addresses the energetics of various intramolecular site-exchange (chain walking) processes and C–H oxidative addition reactions (Scheme 2) of the pentane C–H σ-complexes studied in Chapter 2. Four possible site-exchange processes such as 1,2-, 1,3-, 1,4- and 1,5-migration processes are studied using DFT/M06 level of theory. η2-(H,H)···M type transition states are located for these migrations (Scheme 2). The 1,3-migration is the most favorable process. Two different pentyl hydride isomers, as shown in Scheme 2, are obtained for oxidative addition of methyl and methylene C–H bonds of pentane for all systems, at same level of theory. Oxidative insertion of metal into the methyl C–H bonds is more favorable than insertion into the methylene C–H bonds for all complexes. The activation energies of all site-exchange and C–H oxidative addition processes of the Re(I) complexes are significantly greater than those of the corresponding W(0) complexes. For all these processes, the activation barriers of the electron-deficient NO+ complexes are the greatest among all ligand systems studied, in both Re(I) and W(0) systems. These results are consistent with the experimental results and suggest that the experimentally observed pentyl hydride isomer [(η5–C5H5)Re(CO)(PF3)H(C5H11)] might be Isomer B and not Isomer A (Scheme 2). The C–H oxidative addition reactions are less favorable than dynamic site-exchange processes in all complexes. These results imply that the metal fragments migrate along the pentane chain more easily than insert into the pentane C–H bonds. Scheme 2. Alkane chain walking and C–H oxidative addition reactions Chapter 4 deals with the mechanisms and energetics of a unique metal migration process of an olefin complex that proceeds via olefinic (C–H)···Metal interaction. Migration of the Re(I) fragment from one π face of the olefin to the opposite π face in [(η5–C5H5)Re(NO)(PPh3)(PhCH═CH2)]+ has been documented experimentally by Gladysz and coworkers. The experimental results provide evidences for an intramolecular mechanism for this process (i.e., without styrene dissociation from Re(I)) and based on kinetic isotope effects (KIE), the involvement of a trans C–H bond is indicated. Either oxidative addition or a vinylic (C–H)···Re interaction could account for the experimentally observed kinetic isotope effect. In this study, the free energy of activation for the migration of Re from one enantioface of the olefin to the other through various pathways is computed using DFT calculations at the B3LYP and M06 levels. Two pathways, one that involves migration of Re through a trans (C–H)···Re interaction and another that involves oxidative addition of Re into the trans C–H bond, are identified as possible paths (Scheme 3) at the B3LYP level. Surprisingly, at the M06 level, DFT computes a lower energy path for the conducted tour mechanism that is not consistent with the experimental KIE. But the computed energy profiles for the reaction are consistent with the experiment when computations are carried out at the B3LYP level. Scheme 3. Mechanisms of olefin π face exchange reaction In Chapter 5, the mechanistic studies of C–H metathesis of d6 half-sandwich complex [(η5–C5Me5)Ru(CH3)(CO)(C6H6)] are discussed. A 1-step mechanism that proceeds via a four-center transition state and a 2-step Oxidative Addition and Reductive Coupling mechanism (OA/RC) are identified as possible mechanisms (Scheme 4) using DFT/M06 level of theory. The 1-step mechanism is more favorable than the 2-step mechanism. As in the oxidative addition intermediate, metal–hydrogen bond is observed in the four-center transition state of the 1-step mechanism. This mechanism is referred to as Oxidative Hydrogen Migration (OHM) rather than σ-Bond Metathesis (σ-BM) which proceeds via a transition state without M−H bonding. The effects of metal (M = Fe(II), Ru(II) or Os(II)) and ancillary ligand (L = H–, NHC, CO or NO+) variations on the mechanisms and energetics of the model Cp complex [(η5–C5H5)M(CH3)(L)(C6H6)] are also studied (Scheme 4). Scheme 4. Oxidative hydrogen migration vs Oxidative addition/reductive coupling Increase in the electron-density on the metal center, using electron-donating ligands such as H−, favors the formation of the oxidative species (intermediate or transition state) and reduces the activation barriers of the C–H metathesis reaction. Similarly, the electron-withdrawing NO+ ligand, which reduces the electron density on the metal center, increases the activation energies of the C–H metathesis reaction or disfavors the formation of the oxidative species. Factor affecting the choice of the mechanism of the C–H metathesis reaction is found to be the net charge transfer between the two fragments [(η5–C5H5)M(CH3)(L)] and benzene in [(η5–C5H5)M(CH3)(L)(C6H6)]. The computational studies reported in this thesis provide valuable insight into the mechanisms and energetics of C–H binding, activation and fluxional processes of the (C–H)···Metal σ alkane and alkene complexes. These studies will be helpful in solving problems associated with the C–H activation reactions. Reference Thenraj, M.; Samuelson, A. G. Organometallics 2013, 32, 7141. (For structural formula and figures pl see the abstract pdf file.)

M.Tech. (Department of Chemistry, Faculty of Applied and Computer Sciences), Vaal University of Technology. / Several industries utilize species of synthetic dyes that are found in their wastewater, which is passed out in the environment. Methylene blue is one of the organic dyes that causes water pollution. It causes damage to the aquatic eco-system and health problems to human beings. It is non-biodegradable due to its chemical nature. Advanced oxidation processes (AOP’s) have been developed for the degradation of these dyes, however, some of these methods are limited due to their high cost and low efficiency. Among these methods, Fenton catalysis has been proven to be an effective method due to its low cost, high efficiency, and re-usability. Iron oxide nanoparticles have been mainly used in Fenton process however they are also limitated due to the forming of secondary pollutants, due to catalysts recovery difficulties, hence they require supporting materials. In this work, iron oxide-based catalyst supported on silica nanofibers were fabricated via electrospinning of silica sol incorporated with iron oxide, using three different routes, (a) Method 1 - wetness incipient impregnation, (b) Method 2 - direct addition of iron precursor to the silica sol and (c) Method 3 - incorporation of iron oxide nanoparticles into silica sol. The effect of iron oxide concentration loadings (1 wt%, 2 wt% and 5 wt %) was studied. Increase in iron content resulted in agglomeration of nanoparticles as embedded in the fibers as evident from their SEM images in method 3.1. The SEM results showed diameters from method 1, 2 and 3 ranging from the distribution ranges of 276 – 288 nm, 243 – 265 nm and 188 nm, respectively. EDS showed the presences of Si, P, Fe, O and P. XRD showed a crystalline phase of magnetite (9 nm) and goethite (32 nm) method 1 and 3, with vibrational modes at 3300 cm-1, 1100 cm-1, 950 cm-1 and 580 cm-1 ascribed to O-H, Si-O-Si, Si-O and Fe-O on the FTIR spectra, it showed both the presence of silica and iron oxide. The degradation of methylene blue was monitored by UV-Vis spectroscopy, the Fenton catalytic activity of the iron-oxide supported on silica nanofibers showed higher catalytic activity compared to the unsupported iron-oxide nanoparticles. The catalyst prepared by wetness incipient impregnation (method 1) had a degradation efficiency of 69.1%, the direct addition of iron precursor to the silica sol (method 2) had 75.2% and incorporation of iron oxide nanoparticles magnetite and goethite with the silica sol had 53.7% and 34.7%, respectively. The catalyst prepared by the direct addition of iron precursor in the sol (method 2) showed a high catalytic activity compared to the other catalyst prepared by other methods. Unsupported Iron oxide nanoparticles had a higher degree of leaching of 1.28 ppm magnetite, and 1.68 ppm goethite, compared to the supported iron oxide in method 1 and method 3. The catalyst incorporated with goethite showed a high degree of leaching, 3.95 ppm and 1.33 ppm. The catalyst with high catalytic activity showed a lower degree of leaching with 0.05 ppm.

Neste estudo foi proposto um modelo de formação de biofilme fúngico formado por Candida albicans em dois diferentes substratos associado ou não à glicose para o estudo dos efeitos da terapia fotodinâmica (PDT) utilizando o azul de metileno (AM) como fotossensibilizador, avaliando os efeitos de dois comprimentos de onda distintos incluindo a investigação na captação do AM quando sensibilizados previamente com a glicose. Foi avaliada a susceptibilidade da levedura em suspensão de células crescidas por 24 h (início da fase estacionária) com e sem a adição prévia da glicose. Foram investigadas duas linhagens de C. albicans para a padronização do biofilme, ATCC 90028 e ATCC 10231. Com os biofilmes formados, avaliamos a captação de AM para determinarmos a mais eficiente concentração e tempo de pré-irradiação (PIT). Por último, avaliamos sua sensibilidade à PDT em dois comprimentos de onda distintos (λ = 645 nm e λ = 660 nm) em função do tempo de irradiação (potência = 16 mW, taxa de fluência = 127,3 mW/cm2). A linhagem de célula que possibilitou a formação de biofilme foi a ATCC 10231 em discos de hidrogel. A menor concentração que possibilitou uma melhor captação do AM foi de 500 μM com um PIT de 30 min e em contato prévio com a glicose 50 mM por 90 min. O comprimento de onda mais eficiente, que promoveu redução em leveduras e biofilmes foi o de λ = 660 nm, reduzindo melhor no estudo de leveduras quando sensibilizados com glicose. Nos biofilmes, a redução foi iniciada mais precocemente sem a adição de glicose (com 6 min de irradiação e fluência = 46 J/cm2), porém, em 12 min (fluência = 92 J/cm2) o grupo com glicose passou a ter maior eficiência. / In this study we propose a fungal biofilm model of Candida albicans developed on different substrates for photodynamic effect of photodynamic therapy (PDT) study, using the methylene blue (MB) as photosensitizer, evaluating the effects on two wavelengths distinct, including investigating the difference in uptake of MB when sensitized with glucose. Susceptibility was evaluated in the yeast cell suspension with or without the prior addition of glucose to cells grown for 24 h (early stationary phase). Two strains of C. albicans were investigated for the standardization of the biofilm, ATCC 90028 and ATCC 10231. In biofilms, we evaluated the uptake of MB including cell suspension to determine the most efficient concentration and pre-irradiation time (PIT). Finally, we assess its sensitivity to PDT in two distinct wavelengths (λ = 645 nm and λ = 660 nm) as a function of irradiation time (power = 16 mW, fluence rate = 127,3 mW/cm2). The ATCC strain that allowed the biofilm formation was ATCC 10231 on hydrogel disks. The lowest concentration that enabled better uptake of MB was 500 μM with a PIT 30 min and prior contact with 50 mM glucose f or 90 min. The most effective wavelength, which promoted a reduction in biofilms and yeasts was λ = 660 nm, reducing yeast best when primed with glucose and reduction in biofilms started earlier without the addition of glucose (6 min irradiation, fluence = 46 J/cm2), but at 12 min (fluence = 46 J/cm2) the glucose group now has greater efficiency.

As lesões da periimplantite se não tratadas propriamente podem levar à reabsorção óssea e conseqüente perda do implante. Os implantes com superfície rugosa apresentam melhor resposta do tecido ósseo e melhor ancoragem, entretanto facilitam a formação de biofilme sobre sua superfície, dificultando o tratamento da periimplantite. Diversos métodos de limpeza da superfície do implante têm sido descritos, porém podem danificar sua superfície ou gerar resistência a longo prazo. A terapia fotodinâmica (PDT) surge como uma opção viável e de baixo custo para o tratamento da periimplantite. O objetivo deste estudo in vitro foi analisar a descontaminação bacteriana da superfície de implantes por meio da terapia fotodinâmica. 60 implantes dentários (TiUnite Nobel) foram igualmente divididos (n=10) em 4 grupos e 2 subgrupos. Em G1 não houve descontaminação (controle negativo), enquanto em G2 realizou-se a descontaminação pelo método tradicional com gluconato de clorexidina a 0,12% (controle positivo). G3 (PDT laser+corante) e G4 (sem corante) foram divididos em dois subgrupos cada, sendo que em G3a e G4a utilizou-se a PDT por 3 minutos, G3b e G4b por 5 minutos. Após 5 minutos em contato com o corante azul de metileno a 0,01% (G3), os implantes foram irradiados (G3 e G4) com laser diodo de baixa potência (AsGaAl, 660 nm, 30 mW) por 3 ou 5 minutos (7,2 e 12 J). Após diluições seriadas, os meios de cultura foram mantidos em atmosfera de anaerobiose por uma semana, e então realizada a contagem das unidades formadoras de colônias para cada grupo. Os testes estatísticos de Kruskal-Wallis e Mann-Whitney mostraram haver diferença significante (p<0,001) entre G1 e os demais grupos, bem como G4 comparado com G2 e G3. Melhores resultados de descontaminação foram obtidos para G2 e G3, sem diferença estatisticamente significante entre si. Os resultados obtidos neste estudo sugerem que a terapia fotodinâmica pode ser considerada um método eficaz para redução bacteriana em superfícies de implantes e que somente a irradiação laser, sem a associação do corante, foi menos eficiente (p<0,001) que a PDT. / If periimplantitis lesions are not properly treated, they can lead to bone resorption and loss of the implant. Implants with rough surface present better results and anchorage, however, they can facilitate biofilm formation on their surface, making the treatment of periimplantitis more difficult. Several surface debridement methods have been reported, however, some of them can injury the implant surface or create bacterial resistance in a long term followup. The photodynamic therapy (PDT) is a viable and low-cost option for treatment of periimplantitis. The aim of this in vitro study was to analyze the decontamination of implant surfaces by means of photodynamic therapy. 60 dental implants (TiUnite Nobel) were equally distributed (n=10) into 4 groups and 2 subgroups. At G1 group there was no decontamination (negative control), while in G2 the decontamination was made through the conventional method with chlorhexidine gluconate 0.12% (positive control). G3 (PDT laser+dye) and G4 (without dye) were divided in two subgroups each; while in G3a and G4a PDT was carried out for 3 minutes, G3b and G4b, was for 5 minutes. After five minutes in contact with methylene blue 0,01% dye (G3), the implants were irradiated (G3 and G4) with a diode low-power laser (AsGaAl, 660 nm, 30 mW) for 3 or 5 minutes (7.2 and 12J). After the dilutions, the culture media were kept in anaerobiosis atmosphere for one week, and then the counting of bacteria colonies was done for each group. The statistical tests of Kruskal-Wallis and Mann-Whitney showed significant difference (p<0,001) between G1 and other groups, as well as G4 compared with G2 and G3. Better results of decontamination were obtained in G2 and G3, with no statistical significance between them. The results of this study suggest that the photodynamic therapy can be considered an efficient method of bacteria reduction of implants surfaces, and only the laser irradiation, without dye association, was less efficient (p<0,001) than PDT.

No presente trabalho são apresentadas a síntese e a caracterização estrutural, fotofísica, fotoquímica e fotobiológica de nanopartículas contendo os fotossensibilizadores (FS) Azul de Metileno (AM) e Tionina. AM e Tionina foram incorporados nas nanopartículas sil-AM e sil-Tio pelo processo sol-gel. Nas nanopartículas Cab-Tio, Tionina foi ligada à superfície de sílica CabOsil® através de ligação covalente com reagentes bifuncionais. Todas as nanopartículas mostraram-se esféricas e com de diâmetro médio na faixa de 30 a 60nm. A imobilização dos FS induziu a agregação destes em extensões diferentes para cada tipo de nanopartícula. Foi observado que a maior presença de dímeros de FS leva à menor eficiência de geração de 1O2. Constatou-se que as nanopartículas sofrem pouca influência do meio, uma vez que os FS a elas ligadas não sofreram redução química por NADPH, nem supressão do estado tripleto por íons ascorbato e a supressão de fluorescência por íon brometo foi diminuída. Foi testado também o efeito do recobrimento destas nanopartículas com lipídios dioleilfosfatidil colina (DOPC) e fosfatidilglicerol (PG) e com Polietileno glicol (PEG). A adsorção das nanopartículas sobre membranas miméticas foi reduzida após os recobrimentos, resultado que foi explicado pelas interações de carga superficial (potencial zeta) e pela força de hidratação. As nanopartículas sil-AM e Cab-Tio apresentaram fototoxicidades in vitro, 38% e 20% maiores que os respectivos FS livres. A modificação das nanopartículas de sil-AM com lipídios e com PEG diminuiu a fototoxicidade das mesmas e no caso do recobrimento com lipídios levou ao aumento da toxicidade no escuro. Imagens de microscopia confocal mostraram que as nanopartículas com e sem recobrimento de lipídios entram em células B16. No caso das nanopartículas recobertas, observou-se um perfil de distribuição difuso por todo o citoplasma e no caso de nanopartículas sem recobrimento, estas encontraram-se em poucas regiões vacuolares do citoplasma. O perfil de distribuição homogênea por todo o citoplasma no caso de nanopartículas recobertas com lipídios pode ser o responsável pelo aumento de toxicidade no escuro. Concluiu-se que a ligação dos FS em nanopartículas com diferentes graus de agregação pode ser uma estratégia para obtenção de sistemas com capacidade modulada de geração de 1O2 e com reduzida susceptibilidade às composições do meio. As atividades fototóxicas das nanopartículas contra células B16 mostraram que estas podem ser úteis em Terapia Fotodinâmica de Câncer / In this work we present the synthesis and the characterization (structural, photophysical, photochemical and photobiological) of nanoparticles with incorporated photosensitizers (PS) Methylene Blue (MB) and Thionin. MB and Thionin were incorporated in sil-MB and sil-Th nanoparticles through sol-gel process. In the case of Cab-Th nanoparticles Thionin was linked to the surface of CabOsil® nanoparticles through cross-linking reactions. All nanoparticles were spherical and presented average diameter in the range of 30 to 60nm. Different extension of PS aggregation was observed in each nanoparticle. It was characterized that the higher the proportion of dimers to monomers the smaller the efficiency of singlet oxygen (1O2) generation. It was shown that nanoparticles can protect PS from external interferences, since NADPH did not reduce them, neither were their triplet state quenched by ascorbate ions. Besides, fluorescence quenching by bromide ions was reduced compared to free PS. The effect of covering the nanoparticles with lipids, i.e., di-oleil phosphatidylcholine (DOPC) and phosphatidylglycerol (PG), and with Polyethylene glycol was also tested. The nanoparticle adsorption over membrane mimics was reduced, which was explained by the interaction among surface charges (zeta potential) and by hydration forces. Sil-MB and Cab-Th nanoparticles presented in vitro phototoxicity 38% and 20% higher than the respective free PS. It was observed that the nanoparticle coating with lipids and with PEG reduced their photoxicity. Nanoparticles coated with lipids showed higher toxicity in the dark. Confocal fluorescence images of B16 cells showed that nanoparticles with or without lipid coating enter the cells. In the case of lipid-coated nanoparticles a diffuse distribution profile was observed and in the case of nanoparticles without coating, they concentrated in specific vacuolar regions of the cytoplasm. The homogeneous cytoplasmic distribution profile of lipid-coated nanoparticles can explain the increased toxicity in the dark. It has been concluded that immobilization of PS with different aggregation degrees is a strategy to obtain systems in which the modulated efficiency of 1O2 generation is not affected by the external medium. Finally, based on the observed in vitro phototoxicity activity against B16 cells, these systems can be useful in Photodynamic Therapy of Cancer

Introdução: O transplante de pulmão é uma opção terapêutica bem estabelecida para o tratamento de pneumopatias em estágio final. Uma complicação frequente relacionada aos transplantes é a lesão de isquemia e reperfusão (IR), estando o estresse oxidativo envolvido no processo. O azul de metileno (AM) é um inibidor da produção de espécies reativas de oxigênio, atuando como receptor alternativo de elétrons da xantina oxidase. Objetivo: Avaliar a eficácia do AM como inibidor da lesão de IR em transplante pulmonar de ratos. Métodos: Quarenta ratos fêmea Sprague Dawley (300g - 350g) foram divididas em quatro grupos de dez animais. Os animais foram submetidos a transplante pulmonar unilateral esquerdo. Os enxertos foram expostos a 3 ou 6 horas de isquemia fria seguido de 2 horas de reperfusão. Nos animais do grupo controle, 2 mL de solução salina foram injetados na cavidade peritoneal e, nos animais do grupo experimento, 2 mL de AM a 1% foram injetados da mesma forma. Resultados: A dosagem da PaO2 foi significativamente superior no grupo AM entre os animais submetidos a isquemia de 3 horas (AM = 150,2 ± 50,1 vs Salina = 102,6 ± 40,4 mmHg; p = 0,028), assim como a dosagem do óxido nítrico exalado foi significativamente inferior no mesmo grupo (AM = 3,2 ± 2,0 vs Salina = 5,2 ± 2,3 ppb; p = 0,05). O infiltrado neutrofílico foi menor nos animais do grupo AM submetidos a 6 horas de isquemia no lavado broncoalveolar (LBA); (AM = 11,8 ± 7,4 vs Salina = 30 ± 19,2 X 104/mL; p = 0,023). A análise histopatológica mostrou menor formação de edema perivascular no grupo AM submetido a 6 horas de isquemia (AM = 35,2 ± 7,65 vs Salina = 44,8 ± 6,39%; p = 0,001) e perialveolar no grupo AM submetido a 3 horas de isquemia (AM = 18,4 ± 14,2 vs Salina = 28,1 ± 18,2%; p = 0,041) e menor infiltrado neutrofílico perialveolar na comparação entre os grupos submetidos a 6 horas de isquemia (AM = 2,8 ± 2,2 vs Salina = 5,1 ± 3,1%; p = 0,046). Os níveis de IL-6 no LBA foram inferiores no grupo AM em ambos os tempos de isquemia (3 h - AM = 122,4 ± 24,9 vs Salina = 175.6 ± 50.3 pg/mL; p = 0.008; 6 h - AM = 142 ± 38,7 vs Salina = 351,3 ± 80,7 pg/mL; p = 0,002); Os níveis de TNF-? foram menores no grupo AM submetido a 6 horas de isquemia (AM = 189,5 ± 93,3 vs Salina = 342,9 ± 130,4 pg/mL, p = 0.007). A dosagem do ácido úrico foi significativamente maior no grupo AM em ambos os tempos de isquemia (3 h - AM = 4,7 ± 0,9 vs Salina = 2,7 ± 0,7 mg/dL; p = 0,003; 6 h - AM = 5,3 ± 2,4 vs Salina = 2,3 ± 0,9 mg/dL; p < 0,001). Não houve diferença entre os grupos em relação a apoptose. Conclusão: O AM demonstrou ser uma droga eficaz na prevenção da lesão de isquemia e reperfusão no transplante pulmonar de ratos / Introduction: Lung transplantation (LTx) has become an established therapeutic option for end-stage pulmonary disease. Ischemia reperfusion injury (IR) is a major cause of organ dysfunction after LTx and oxidative stress is involved in this process. Methylene blue (MB) is an inhibitor of reactive oxygen species production. Objective:To investigate the effects of MB on the IR in a rodent model of LTx. Methods: Forty female Sprague Dawley rats (300g - 350g) were divided into four groups (n = 10) according to treatment (saline solution or MB) and graft cold ischemic time (3 or 6 hours). Animals underwent a left-sided unilateral lung transplantation. Recipients received 2mL of intraperitoneal saline or MB 1%. After 2 hours of reperfusion, animals were killed and blood gas, exhaled nitric oxide, cell count and cytokines leves in bronchoalveolar lavage (BAL) as well as histopathology, serum uric acid and apoptosis were evaluated. Results: PaO2 was significantly higher in MB group undergoing 3 hour ischemic time (MB =150.2 ± 50.1 vs Saline = 102.6 ± 40.4 mmHg; p = 0.028). Exhaled nitric oxide values showed differences only between groups with 3 hour of ischemia (MB = 3.2 ± 2.0 vs Saline = 5.2 ± 2.3 ppb; p = 0.05). Neutrophils in BAL were different between groups subjected to 6 hours of ischemia (MB = 11.8 ± 7.4 vs Saline = 30.0 ± 19.2 x 104/mL; p = 0.023). IL-6 levels in BAL were lower in MB group in both ischemic time (3 h - MB = 122.4 ± 24.9 vs Saline = 175.6 ± 50.3 pg/mL; p = 0.008; 6 h - MB = 142 ± 38.7 vs Saline = 351.3 ± 80.7 pg/mL; p = 0.002); TNF-? levels were also lower in MB group undergoing 6 hour of ischemia (MB = 189.5 ± 93.3 vs Saline = 342.9 ± 130.4 pg/mL; p = 0.007). The number of neutrophils in lung parenchyma were reduced in MB group (6 h - MB = 2.8 ± 2.2 vs Saline = 5.1 ± 3.1%; p = 0.046) and also decreased edema in perivascular (6 h - MB = 35.2 ± 7.65 vs Saline = 44.8 ± 6.39%, p = 0.001) and perialveolar tissues (3 h - MB = 18.4 ± 14.2 vs Saline = 28.1 ± 18.2% p = 0,041) were observed. Uric acid levels were higher in MB group in both ischemic time (3 h - MB = 4.7 ± 0.9 vs Saline = 2.7 ± 0.7 mg/dL; p = 0.003; 6 h - MB = 5.3 ± 2.4 vs Saline = 2.3 ± 0.9 mg/dL; p < 0.001).There were no difference in the expression of apoptosis. Conclusion: MB was able to prevent ischemia-reperfusion injury in this lung transplantation model and represent a new option for further studies