Made available in DSpace on 2014-10-09T12:42:50Z (GMT). No. of bitstreams: 0 / Made available in DSpace on 2014-10-09T14:07:27Z (GMT). No. of bitstreams: 1 06112.pdf: 5630844 bytes, checksum: d96c90e096947d42a20110e42e3cc1e8 (MD5) / Dissertacao(Mestrado) / IPEN/D / Instituto de Pesquisas Energeticas e Nucleares - IPEN/CNEN-SP

Orientador: Marcelo Giannini / Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Odontologia de Piracicaba / Made available in DSpace on 2018-08-13T10:19:14Z (GMT). No. of bitstreams: 1 Oliveira_MarceloTavaresde_D.pdf: 15233943 bytes, checksum: f6125d2132f3fd48d81be1d3cda778aa (MD5) Previous issue date: 2009 / Resumo: O uso de técnicas alternativas para preparo cavitário vem ganhando grande importância na Odontologia moderna. Entre estas tecnologias, o laser tem sido alvo de inúmeros estudos e para a realização de preparos cavitários o laser de Er:YAG é um dos mais indicados. Os objetivos deste estudo foram avaliar os efeitos de diferentes parâmetros de irradiação do substrato dentinário com laser de Er:YAG nos procedimentos restauradores adesivos e suas conseqüências sobre o substrato e sobre a união resina-dentina. Os testes foram realizados em superfícies dentinárias planificadas de terceiros molares humanos hígidos unidos a três diferentes sistemas adesivos (Single Bond 2 - 3M ESPE, Clearfil Protect Bond - Kuraray Medical Inc. e Clearfil Tri-S Bond - Kuraray Medical Inc.). Estas avaliações compreenderam o estudo da permeabilidade dentinária, da resistência de união, da longevidade e qualidade da união, além da análise da ultra-estrutura da interface de união através de microscopia eletrônica de transmissão e microscopia de varredura confocal laser. Os dados de permeabilidade e resistência de união foram analisados e submetidos à análise estatística com ANOVA 1-critério e ANOVA 3-critérios, respectivamente; ao encontrar diferenças significativas, o teste de Tukey foi utilizado. O nível de significância foi fixado em 5%. Os resultados mostraram ocorrer uma redução significativa na permeabilidade dentinária e que o parâmetro de 120mJ/4Hz foi o que promoveu a maior redução. Quanto à resistência de união, este estudo demonstrou que inicialmente, a irradiação com laser de Er:YAG reduz os valores de resistência de união, no entanto após 6 meses os valores são semelhantes tanto para o grupo controle, quanto para os grupos que foram irradiados. Além disso, pôde ser observado, que os sistemas autocondicionantes foram menos susceptíveis aos efeitos da irradiação do substrato, apresentando um comportamento mais uniforme. O sistema autocondicionante de dois passos também se mostrou mais estável ao longo do tempo. As imagens em microscopia eletrônica de transmissão e microscopia de varredura confocal laser, mostraram a formação de camada híbrida para todos os adesivos estudados com diferentes espessuras, quando aplicados sobre a dentina abrasionada. Não foi possível identificar camada híbrida uniforme e bem definida quando os sistemas foram aplicados em dentina irradiada com laser de Er:YAG, independentemente do parâmetro utilizado. Diante disso, os resultados sugerem que a irradiação da dentina com laser de Er:YAG reduz a permeabilidade dentinária, podendo influenciar na união dos adesivos à dentina nos períodos iniciais, entretanto após 6 meses os grupos irradiados foram iguais ao grupo controle. O exame ao microscópio revelou que a formação da camada híbrida é irregular e não-uniforme quando a dentina foi irradiada. / Abstract: The use of alternative techniques for cavity preparation is more common nowadays. Among these technologies, laser is the target of a great number of studies. The objectives of this study were to evaluate the effects of different parameters of dentin irradiation with Er:YAG laser on the adhesive procedures and their consequences over the substrate and resin dentin interface. Procedures were performed over flat dentin surfaces of human third molars bonded to three different adhesive systems ((Single Bond 2 - 3M ESPE, Clearfil Protect Bond - Kuraray Medical Inc. e Clearfil Tri-S Bond - Kuraray Medical Inc.). The evaluation tests were: dentin permeability, microtensile bond strengths, durability and the analysis of the ultra-structure using transmission electron microscope (TEM) and confocal laser scanning microscope (CSLM). Dentin permeability data and microtensile bond strength data were analyzed with one-way ANOVA and three-way ANOVA, respectively. Tukey test was used to point out differences among the tested groups. The significance level was fixed at 5%. Results showed a significative reduction on the dentin permeability and the 120mJ/4Hz parameter promoted the greatest reduction. For the microtensile bond strength test, this study showed that the irradiation with Er:YAG laser reduce the bond strength values at the baseline. After six months all groups became similar. Furthermore, self-etching systems were less affected by the Er:YAG laser irradiation. The two-step self-etching system seemed the more stable over time. TEM and CLSM images showed the hybrid-layer formation for all adhesive tested. The hybrid-layers had different thicknesses when the adhesive systems were bonded to abraded dentin. The observation of a uniform and well defined hybrid-layer was not possible when the adhesive systems were bonded to laser irradiated dentin, regardless of the used parameter. According to the data presented, dentin irradiation with Er:YAG laser reduce the permeability of the substrate, interfering on the bonding strength to dentin at the baseline, however, after six months laser irradiated groups were similar to control groups. Microscope analysis revealed that hybrid layer formation was irregular and non-uniform when dentin was laser irradiated. / Doutorado / Doutor em Materiais Dentários

Os primatas neotropicais, ou primatas do novo mundo, são pertencentes a um grupo diverso e variado de animais, com características anatômicas, comportamentais e taxonômicas próprias, que os diferem dos primatas do velho mundo. Estima-se que representem 31% dos primatas não-humanos do planeta. Têm sofrido com as altas taxas de crescimento populacional e perturbação antrópica resultante do desflorestamento, agricultura e criação de gado. Com o intuito de reverter esta situação e maximizar o potencial reprodutivo das espécies, o desenvolvimento de biotecnologias associadas à reprodução tem sido amplamente justificado nos dias de hoje, por representarem importantes ferramentas reprodutivas, contribuindo para a preservação da biodiversidade e manutenção da diversidade genética. Assim, o objetivo do presente estudo foi padronizar a colheita de sêmen de Alouatta guariba clamitans por eletroejaculação por via retal com posterior análise do sêmen fresco, procedimentos nunca descritos para a espécie, e adaptar o método de preparo de sêmen de Leontopithecus chrysomelas, Alouatta caraya e Alouatta guariba clamitans para microscopia eletrônica de transmissão, permitindo a comparação da ultraestrutura dos espermatozoides das três espécies e apresentando resultados também inéditos na literatura científica. Para este estudo, foram utilizados quatro exemplares de L. chrysomelas, dois exemplares de A. caraya e dois exemplares de A.g. clamitans, machos adultos mantidos em cativeiro. L. chrysomelas tiveram o sêmen colhido por vibroestimulação peniana, e A. caraya e A.g. clamitans foram submetidos à eletroejaculação por via retal. Após a obtenção dos ejaculados, o sêmen foi avaliado quanto ao volume, pH, concentração, motilidade, integridade de membrana plasmática, integridade de membrana acrossomal e morfologia, e posteriormente preparado para microscopia eletrônica de transmissão. A análise dos 12 espermatozoides das três espécies estudadas revelou que todos apresentam os componentes básicos descritos para o espermatozoide humano cabeça, colo e cauda, formada pelas peças intermediária, principal e terminal com variações na forma e tamanho de cada componente. As diferenças mais significativas encontradas entre os espermatozoides comparados foram extensão de acrossomo, comprimento, largura e organização de peça intermediária. O presente estudo atingiu os objetivos propostos, apresentando resultados inéditos na literatura científica / The neotropical primates, also known as primates of the new world, are a diverse and varied group of animals, with unique anatomical and taxonomic characteristics, as well a unique behaviour, that distinguish those from the old world primates. It is estimated that these present 31% of the non-human primate population of the planet. High taxes of populational growth and anthropological disturbance have been undergoing due to deforestation, agriculture and cattle farms. In order to revert this situation and maximize the reproductive potential of the species, the development of biotechnologies associated to the reproduction are widely justified nowadays, for representing important reproductive tools, contributing to the preservation of the biodiversity and maintenance of genetic diversity. Hence, the purpose of this study was to collect Alouatta guariba clamitans semen by rectal electroejaculation then analyze the fresh semen, procedures never before described for the species, and adapt the preparation method of Leontopithecus chrysomelas, Alouatta caraya and Alouatta guariba clamitans semen for electron transmission microscope, allowing the comparison of the spermatozoa ultrastructure from the three species and also presenting new results to the scientific literature. For this study, it was used four specimens of L. chrysomelas, two specimens of A. caraya and two specimens of A.g. clamitans, adult males kept in captivity. L. chrysomelas had the semen taken by penine vibratory stimulation, and A. caraya and A.g. clamitans were submitted to rectal electroejaculation. After the semen was obtained, it was evaluated regarding the volume, pH, concentration, motility, sperm membrane integrity, acrosome integrity and morphology, and then prepared for electron transmission microscopy. The analysis of the spermatozoa of the three studied species revealed that all present the basic components described for the human spermatozoa - head, neck region and tail, formed by middle, principal and terminal pieces - with variations in 14 shape and size of each component. The most significant differences found between the compared spermatozoa were the acrosome extension and middle piece length, thickness and organization. The present study attained the proposed goals, presenting unprecedented results for the scientific literature

Orientador: Léa Rodrigues Simioni / Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Ciências Médicas / Made available in DSpace on 2018-08-19T21:13:03Z (GMT). No. of bitstreams: 1 Ferreira_SandroRostelato_D.pdf: 2167840 bytes, checksum: f1dd1c4722a358caf196b8ef326c3b3f (MD5) Previous issue date: 2012 / Resumo: Rhinella schneideri, conhecido previamente como Bufo paracnemis, é um sapo comum em muitas regiões do Brasil. O veneno destas espécies exerce importante efeito cardiovascular em humanos e animais, mas pouco se sabe sobre sua atividade neuromuscular. Neste trabalho, nós avaliamos a neurotoxicidade do veneno de R. schneideri em preparações neuromusculares de pintainho e camundongo. Através da compressão manual das glândulas parótidas localizadas atrás dos olhos, coletou-se a secreção e então realizada a extração com metanol. O extrato metanólico foi liofilizado e testado em preparações biológicas. Preparações biventer cervicis (BC) de pintainho e nervo frênico diafragma (NFD) de camundongo foram utilizadas para o registro miográfico através de estimulação elétrica indireta para medidas eletrofisiológicas, análise morfológica e microscopia eletrônica de transmissão. Frações ativas do extrato metanólico foram obtidas submetendo-se à coluna de fase reversa Luna PFP (250 x 4,6 mm). O extrato metanólico (50 ?g/ml) causou somente facilitação da neurotransmissão em preparações NFD. Ao contrário, causou bloqueio neuromuscular significativo em preparações BC que foram concentração-dependente (3, 10 and 30 ?g/ml; a 37º C) com tempo para 50% de bloqueio, média ± erro padrão: 84±10, 51±3 e 12±0,8 min com 3, 10 e 30 ?g/ml, respectivamente; n=6-8 cada, precedido por facilitação da neurotransmissão. Não houve inibição significativa das respostas contraturantes à ACh (110 ?M) ou KCl (20 mM) após bloqueio completo em qualquer concentração testada. Em preparações BC incubadas com o extrato metanólico (10 ?g/ml) a 22º C por 70 min não observou-se qualquer alteração das respostas musculares (117±3%; n=5), mas quando a temperatura do banho foi elevada a 37º C, 50% de bloqueio ocorreu após 92±3 min (n=5; p<0.05). A incubação de preparações BC curarizadas (d-Tc, 1 ?g/ml) com o extrato metanólico (10 ?g/ml) resultou em completo e irreversível bloqueio enquanto que as preparações tratadas somente com curare mostraram a reversão completa da resposta contrátil após várias lavagens. Não houve aumento significativo nos níveis de liberação de creatinoquinase (90±21 vs. 80±15 U/l, antes e após 120 min de incubação com o extrato, respectivamente, n=5) além da ausência de alterações na morfologia das fibras musculares ou na porcentagem de danos na fibra (2.4±0.9 vs. 2.3±0.5 %, antes e após 120 min de incubação com o extrato, respectivamente, n=5). O extrato metanólico (50 ?g/ml) aumentou a resposta contrátil mas não alterou o potencial de membrana em repouso (-81±1 mV e -78±1 mV para controle e preparação tratada após 60 min). Registros eletrofisiológicos mostraram que houve um aumento progressive na frequência dos potenciais de placa terminal em miniatura (PPTM) de 34±3,5 (controle) para 88±15 (após 60 min de incubação com o extrato); houve também um aumento nos valores do conteúdo quântico, de 128±13 (controle) para 272±34 e 171±11 após 5 min e 60 min, respectivamente, em preparações tratadas com o extrato metanólico. A microscopia eletrônica de transmissão mostrou que o volume ocupado pelas vesículas sinápticas foi significativamente reduzida (32±5%; p<0.05) após 5 min mas este efeito foi reversível após 60 min de incubação para as preparações tratadas com 50 ?g/ml do extrato metanólico. Não houve dano estrutural distinguível na membrana do terminal nervoso e nas mitocôndrias das preparações tratadas com o extrato, quando comparada com as preparações controle. O pré-tratamento das preparações NFD com ouabaína (1 ?g/ml), um inibidor da bomba de Na+/K+-ATPase, por 5 min antes da incubação com o extrato, preveniu o aumento do conteúdo quântico comparado com preparações controle (118±18, 117±18 e 154±33 para preparações controle-ouabaína e tratadas com ouabaína e incubadas com o extrato por 5 min e 60 min, respectivamente). A cromatografia por HPLC do extrato metanólico resultou em 24 frações, das quais 4 (frações 20, 21, 22 e 24) causaram bloqueio neuromuscular em preparações BC. A fração 20 (3 ?g/ml) foi escolhida por ser 3 vezes mais potente que as demais e causou bloqueio neuromuscular significativo (p<0.05; tempo para 50% de bloqueio: 43±4 min; n=4) precedido por facilitação em preparações BC a 37º C. A fração 20 não inibiu as respostas contraturantes à ACh (110 ?M) ou KCl (20 mM) após completo bloqueio neuromuscular em preparações BC. Em preparações NFD, a fração (15 ?g/ml) aumentou significativamente os valores do conteúdo quântico de 117±18 (controle) para 236±44 após 5 min de incubação (n=4; p<0.05). Estes resultados indicam que o extrato metanólico do veneno de R. schneideri é capaz de interferir com a neurotransmissão por ativar e/ou bloquear a liberação da acetilcolina nos sítios pré-sinápticos, provavelmente envolvendo a bomba de Na+-K+-ATPase, sem causar qualquer dano à musculatura / Abstract: Rhinella schneideri, previously known as Bufo paracnemis, is a common toad in many regions of Brazil. The venom of this species exerts important cardiovascular effects in humans and animals, but little is known of its neuromuscular activity. In this work, we examined the neurotoxicity of R. schneideri venom in chick and mouse neuromuscular preparations. Venom was collected by manual compression of the large parotid glands behind the eyes and then extracted with methanol. The extract was lyophilized prior to testing in biological preparations. Chick biventer cervicis (BC) and mouse phrenic nerve-diaphragm (PND) preparations were mounted for conventional twitch-tension recording in response to indirect stimulation, for electrophysiological measurements, morphological analysis and transmission electronic microscope. Also, an active fraction of this methanolic extract obtained by reverse phase HPLC on a Luna PFP (250 x 4.6 mm) column. The methanolic extract (50 ?g/ml) caused facilitation but no neuromuscular blockade in PND preparations. In contrast, significant (p<0.05) concentration-dependent (3, 10 and 30 ?g/ml) neuromuscular blockade (time for 50% blockade, mean±S.E.M.: 84±10, 51±3 and 12±0.8 min with 3, 10 and 30 ?g/ml, respectively; n=6-8 each) preceded by facilitation was seen in BC preparations at 37oC. There was no inhibition of contractures to exogenous ACh (110 ?M) or KCl (20 mM) after complete blockade by any of the concentrations tested. Incubation of BC preparations with methanolic extract (10 ?g/ml) at 22oC for 70 min did not affect neuromuscular transmission (117±3%; n=5), but when the bath temperature was increased to 37oC, 50% blockade occurred within 92±3 min (n=5; p<0.05). Incubation of curarized (d-Tc, 1 ?g/ml) BC preparations with methanolic extract (10 ?g/ml) resulted in complete, irreversible blockade whereas preparations treated with curare alone showed complete reversion in the twitch-tension after washing. There was no significant increase in creatine kinase levels (90±21 vs. 80±15 U/l, before and after a 120 min incubation with extract, respectively; n=5) and no significant alterations in muscle fiber morphology or in the percentage of damaged fibers (2.4±0.9 vs. 2.3±0.5 % before and after a 120 min incubation with extract, respectively; n=5). The methanolic extract (50 ?g/ml) increased the twitch-tension but did not alter the membrane resting potential (-81±1 mV and -78±1 mV for control and poison-treated preparations after 60 min). Electrophysiological measurements showed that there was a progressive increase in the frequency of miniature end-plate potentials (MEPPs) from 34±3.5 (control) to 88±15 (after a 60 min incubation with extract); there was also an increase in the end-plate potentials (based on the quantal content) from 128±13 (control) to 272±34 and 171±11 after 5 min and 60 min, respectively, in extract-treated preparations. TEM showed that the fractional volume occupied by synaptic vesicles was significantly reduced (32±5%; p<0.05) after a 5 min but this effect was reversible after 60 min of incubation to 50 ?g/ml of methanolic extract. There was no structural damage to the membrane of the terminal boutons and the mitochondria of extract-treated preparations were indistinguishable from those of control preparations. Pretreatment of the preparations with ouabain (1 ?g/ml), a Na+/K+-ATPase pump inhibitor, for 5 min prior to incubation with methanolic extract prevented the increase in quantal content compared to preparations without extract (118±18, 117±18 and 154±33 for ouabain-treated controls and ouabain-treated preparations incubated with venom for 5 min and 60 min, respectively). HPLC of the methanolic extract resulted in 24 fractions, of which four (fractions 20, 21, 22 and 24) produced blockade in BC preparations. Fraction 20 (3 ?g/mL) was chosen because was the most potent of the four fractions and caused significant (p<0.05) neuromuscular blockade (time for 50% blockade: 43±4 min; n=4; mean±SEM) preceded by facilitation in BC preparations at 37oC. Fraction 20 did not inhibit contractures to exogenous ACh (110 ?M) or KCl (20 mM) after complete neuromuscular blockade in BC preparations. In PND preparations, fraction 20 (15 ?g/mL) significantly increased the quantal content value from 117±18 (control) to 236±44 after 5 min (n=4; p<0.05). These results indicate that the methanolic extract of R. schneideri is capable to interfere with the neurotransmission by activing and/or blocking the pre-synaptic acetylcholine release by an activity involving the Na+-K+-ATPase pump, without damaging the muscle membrane / Doutorado / Farmacologia / Doutor em Farmacologia

Magister Scientiae (Medical Bioscience) - MSc(MBS) / Despite the advances in therapies such as chemotherapy and radiotherapy, tumours have been shown to be resistant to the treatments. Gold nanoparticles (AuNPs) have been recognized as effective radiosensitizers of low energy (e.g. 200–500 kV) X-rays, leading to the emission of Auger electrons that cause highly localised ionizing damage to cells. Spherical AuNPs were synthesised via the reduction of the chloroaurate ions by sodium citrate. Characterisation of AuNPs involved UV-visible spectrophotometry, zeta (Z) potential, dynamic light scattering (DLS) and polydispersity index (PDI) measurements for determination of surface plasmon resonance (SPR), surface charge and stability, as well as transmission electron microscopy (TEM) for hydrodynamic core sizes, size distribution width and shape of AuNPs. Both the 5 and 10 nm AuNPs were found to be anionic with λmax absorbance of 525 nm and uniform size distribution. DLS measurement at 38.12 nm and 48.50 nm, respectively for 5 nm and 10 nm AuNPs, points to aggregation of the AuNPs. However, TEM measurements confirmed the core size of the 10 nm AuNPs. Non-malignant Chinese hamster ovary (CHO-K1), brain endothelial (BEnd5), breast (MCF-10A), isolated human lymphocytes and malignant breast (MCF-7) cell lines were treated with 50 μg/ml of AuNPs, and irradiated with either 1, 2 or 4 Gy X-rays or 1 or 2 Gy p(66)/Be neutron radiation. The γ-H2AX foci assay, cytokinesis-block micronucleus assay, MTT assay and fluorescence-activated cell sorting (FACS) was used to determine that amount of double stranded breaks (DSBs) in isolated lymphocytes, the presence and number of micronuclei (MNi) within binucleated cells (BNCs), cell viability and cell cycle progression, respectively. Preliminary experiments that established the reliability of the study regarding the induction of DNA damage after the bombardment of AuNPs by scattered low kV X-rays, were carried out on lymphocytes. Combined treatment (AuNPs and radiation) resulted in more endogenous foci in comparison to lymphocytes that were treated with AuNPs only. The CHO-K1 and MCF-7 cells showed higher MNi frequencies after the combination treatment of AuNPs and radiation compared to the number of MNi in samples exposed to AuNPs and radiation separately. The AuNPs alone influenced the cellular kinetics of all cell types. Interaction indices, which is the enhancement factor of AuNPs in combination with radiation, for AuNPs and 6 MV 2 Gy X-rays of 1.6 to 1.7 and 1.3 to 1.4 have respectively been determined for CHO-K1 and MCF-7 cells, whilst that for the other cell types used in the study were not different from Unity. As expected, the interaction indices between AuNPs and p(66)/Be neutrons was lower than the interaction indices after 2 Gy X-rays, as p(66)/Be neutrons interact only with the nuclei of the AuNP's atoms and the X-ray photons interact with the orbital electrons of the atoms of the AuNPs leading to Auger electron emission. The cell viability assay showed that 50 μg/ml of AuNPs had an inhibitory effect on cellular proliferation, in all four cell linnes whereas the lower concentrations (2.5, 5 and 10 μg/ml) had no effect. Results in this study, revealed an increase in the accumulation of CHO-K1 an MCF-7 cells in the G₂/M phase of the cell cycle after being treated with AuNPs followed by X-ray radiation, suggesting that the cells have possibly been sensitised to the damaging effects of radiation. Further studies are required to quantify internalised AuNPs and to then link the possible concentration differences of the AuNPs to differences in radiation damage effects observed for the different cell types.

The small intestine mucosa is known to be able to adapt itself to several kinds of both physiological and pathological conditions. The adaptive patterns of the structure of the enterocytes, particularly their apical surface (brush border), were studied in three models: (1) in rats, subjected to antrectomy or antral exclusion, combined with gastroduodenostomy and gastrojejunostomy; (2) in rats with alloxan dia­betes; (3) in children with coeliac disease; a) in its active phase; b) after long-term treatment with gluten-free diets; c) after long-term challenge with dietary gluten following treatment; d) after short-term elimination of dietary gluten. Gut mucosa from fasting or fed, normal or sham-operated rats, fasting cats, and short-statured children with no signs of gastrointestinal disease served as controls. - The specimens were prepared for light microscopy (LM), scanning electron microscopy (SEM), and transmission electron microscopy (TEM). Quantitation of structural variables was achieved by means of LM and TEM morphometrical procedures. Differentiation of the rat enterocytes from the base to the crest of the villi was structurally reflected by doubling of their apical cell area, an increase in cell height, and a decrease of both nuclear and mitochondrial volume densities. In mature normal rat enterocytes, high-power SEM showed regularly arranged, nude microvilli in thir apical surfaces, whereas in cat and man the apical surfaces were covered by a thick glycocalyx. - Fasting for 24 hours decreased the total length of the rat small intestine and the height of the enterocytes. Antrectomy and antral exclusion with gastrojejunostomy produced an increase of the apical surfaces of the enterocytes of the seif-emptying duodenal blind loop, whereas no changes occurred after antrectomy with gastroduodeno­stomy. In the jejunum, the apical surface area was reduced both after antrectomy and antral exclusion. In the diabetic rats a slight decrease of the apical surface area, together with an elongation of both the vil­li and the crypts, was observed in the jejunum, whereas no structural changes occurred in the duodenal mucosa. Both in active coeliac disease and after long-term challenge with dietary gluten, SEM analyses showed uniformly destructed villi. The api­cal surfaces of the enterocytes were frequently convex and irregular in size and delineation (the surface of the normal enterocytes was polygo­nal and flat). Ultrastructurally, the apical surfaces were severely damaged with a distortion of the glycocalyx and with marked irregularity of the microvilli. - After gluten elimination, the surface ultrastructu­re of the enterocytes in the coeliac gut mucosa generally showed a rapid, clear-cut restoration despite a remaining severe atrophy of the villi. Successful dietary treatment (after about one year of gluten-free diet) restored the small intestine mucosa to normal as assessed both by LM and low-power SEM. In contrast, high-power SEM often disclosed per­sisting lesions of the enterocytes. Another provocation with gluten for up to 9 days in clinically healed coeliac mucosa did not significantly alter the surface ultrastructure of the enterocytes. / <p>S. 1-52: sammanfattning, s. 53-138: 5 uppsatser</p> / digitalisering@umu

The thesis reports on fabricating alternative solid phase extraction (SPE) sorbents and colorimetric probes based on electrospun nanofibers for alkylphenols (APs). Hydroxyl methylated styrene [poly(co-styrene-CH₃OH)] and 3-oxobutanoate styrene [poly(co-styrene-OCOCH₃COCH₃)] copolymers were synthesized and fabricated into sorbent materials by electro-spinning/spraying. The fabricated morphologies consisting of bead free fibers, beaded fibers and particles were evaluated as SPE sorbents using batch experiments. Electropun fibers proved to be better sorbents as they exhibited extraction efficiency that exceeded 95% compared to 60% for beaded fibers and 40% for particles. In view to reduce sample and solvent volumes, smooth fibers were packed into pipette tips as SPE devices that yielded quantitative recoveries of APs from spiked wastewater samples. Recoveries ranged from 70% to 125% with LOD of 0.008, 0.01 and 0.1 μg mL⁻¹ for 4-tert octylphenol (4-t-OP), 4-octylphenol (4-OP) and 4-nonylphenol (4-NP) respectively, when using high performance liquid chromatography-fluorescence detector (HPLC-FLD). Furthermore, amino functionalised polydiacetylene polymers (PDAs), citrate capped gold (AuNPs) and silver nanoparticles (AgNPs) were evaluated as colorimetric probes for visual detection of APs. In colloidal studies, AuNPs probe showed a colour change from wine red to green upon introduction of analyte. UV-vis spectroscopy revealed the shifting of the surface plasmon resonance (SPR) peak from 525 nm to 729 nm induced by aggregation of AuNPs. For AgNPs probe, a colour change was observed from yellowish green to brown. Transmission electron microscopy (TEM) studies showed growth of AgNPs. A presumed oxidation of the analyte, forming an absorbing compound at 279 nm in both AgNPs and PDAs probes was also observed. For PDAs probe the colour change was from purple to pink. Concentrations as low as 30 μg mL⁻¹ were detectable in all colloidal based probes. Further colorimetric investigations were conducted with electrospun AuNPs-nylon 6 fiber mat. A colour change from purplish red to navy blue at concentrations of 1000 μg mL⁻¹ was observed. Electrospun AgNPs –nylon 6 fiber mat did not show a distinct colour change. High resolution scanning electron microscopy (HRSEM) revealed the analyte inducing the assembly of AuNPs and AgNPs as they covered the surface of the nanofiber mat. Electrospun nanofibers are a platform for analysis and thus tuning their chemistry will lead to sensitive and selective methods

The aim of this thesis is to investigate the methodology of three dimensional chemical imaging of nanoparticles through the use of scanning transmission electron microscope (STEM) – energy dispersive X-ray (EDX) spectroscopy. In this thesis, an absorption correction factor is derived for spherical nanoparticles that can correct X-ray absorption effects. Quantification of EDX spectra of nanoparticles usually neglects X-ray absorption within the nanoparticle but may lead to erroneous results, thus an absorption correction is important for accurate compositional quantification. The absorption correction presented is verified through comparison with experimental data of Au X-ray peaks in spherical Au nanoparticles and is found to agree excellently. This absorption correction allows accurate compositional quantification of large ( > 100 nm) particles with STEM-EDX.Three dimensional chemical mapping is achievable through the use of EDX spectroscopy with electron tomography. Here, the methodology of STEM-EDX tomography is fully explored, with a focus on how to avoid artefacts introduced through detector shadowing and low counts per pixel. A varied-time acquisition scheme is proposed to correct for detector shadowing that is shown to provide a more constant intensity over a series of projections, allowing a higher fidelity reconstruction. The STEM-EDX tomography methodology presented is applied to the study of AgAu nanoparticles synthesized by the galvanic replacement reaction. The elemental distribution as a function of the composition of the as-synthesized nanoparticles is characterised and a reversal in the element segregated to the surface of the nanoparticles is found. The composition at which the reversal takes place is shown to correlate with a peak in the catalytic yield of a three component coupling reaction. It is hypothesized that a continuous Au surface results in the optimum catalytic conditions for the reaction studied, which guides the use of galvanically prepared AgAu nanoparticles as catalysts.

Graphene, a one-atom thick sheet of carbon, is the thinnest, strongest and most electrically conductive material ever discovered. Alongside carbon nanotubes it is part of the group of nanocarbons whose unique properties have sparked huge interest in possible applications, including electronic devices, solar cells and biosensors. Doping of these materials allows for the modification of their optical and electronic properties,which is crucial to realising these applications. Studying the properties of these doped materials at atomic resolution and finding controllable and industrially scalable routes to doping, such as low energy ion implantation, are thus essential if they are to becomethe materials of the future. In this thesis, highly localised optical enhancements in metal doped graphene are studied using energy-filtered transmission electron microscopy in a monochromated and aberration corrected electron microscope. The ideal conditions for imaging the low energy loss region of graphene using EFTEM are discussed and new methods to compensate for image artifacts when using this technique at high resolution are presented. Density functional theory is used to reveal new visible spectrum plasmon excitations in the electron energy loss spectra of boron and nitrogen doped nanocarbons. Atomic resolution scanning transmission electron microscopy and nanoscale electron energy loss spectroscopy are used to investigate controllable and defect-free substitutional doping of suspended graphene films through low energy ion implantation. Computational methods for filtering high angle annular dark field images are shown and software for the automated processing and spectroscopic analysis of these images is developed.

A number of investigators have reported enhancement in oxidation and wear resistant of carbon-carbon composites (CCC) in the presence of protective coating layers. However, application of a surface and subsurface coating system that can preserve its oxidation and wear resistance along with maintaining lubricity at high temperature remains unsolved. To this end, thermodynamically stable protective oxides (ZnO/Al2O3/ZrO2) have been deposited by atomic layer deposition (ALD) to infiltrate porous CCC and graphite foams in order to improve the thermal stability and wear resistance in low and high speed sliding contacts. Characterization of microstructural evolution was achieved by using energy dispersive x-ray spectroscopy (EDS) mapping in scanning electron microscope (SEM) coupled with focused ion beam (FIB), x-ray tomography, high resolution transmission electron microscopy (HRTEM), scanning transmission electron microscopy (STEM) and X-ray diffraction (XRD). Evaluation of the tribological properties of CCC coated with abovementioned ALD thin films were performed by employing low speed pure sliding tribometer and a high speed/frequency reciprocating rig to simulate the fretting wear behavior at ambient temperature and elevated temperatures of 400°C.It was determined with x-ray tomography imaging and EDS mapping that ALD ZnO/Al2O3/ZrO2 nanolaminates and baseline ZrO2 coatings exhibited excellent conformality and pore-filling capabilities down to ~100 μm and 1.5 mm in the porous CCC and graphite foam, respectively, which were dependent on the exposure time of the ALD precursors. XRD and HRTEM determined the crystalline phases of {0002} textured ZnO (wurtzite), amorphous Al2O3, and {101}-tetragonal ZrO2. Significant improvements up to ~65% in the sliding and fretting wear factors were determined for the nanolaminates in comparison to the uncoated CCC. A tribochemical sliding-induced mechanically mixed layer (MML) was found to be responsible for these improvements. HRTEM confirmed the presence of a high density of ZnO shear-induced basal stacking faults inside the wear tracks responsible for intrafilm shear velocity accommodation that mitigated friction and wear.