Wong Hau Yan Connie. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2000. / Includes bibliographical references (leaves 86-95). / Abstracts in English and Chinese. / Abstract --- p.ii / Acknowledgements --- p.ix / Contents --- p.x / List of Figures --- p.xiii / List of Tables --- p.xv / Abbreviations --- p.xvi / Chapter Chapter 1: --- Introduction / Chapter 1.1 --- Role of extracellular nucleotides in equine sweat gland epithelial cells --- p.1 / Chapter 1.2 --- Subdivision of P1 and P2 purinoceptor --- p.4 / Chapter 1.3 --- General properties of P2 purinoceptor --- p.5 / Chapter 1.3.1 --- P2X purinoceptor family --- p.5 / Chapter 1.3.2 --- P2Y purinoceptor family --- p.8 / Chapter 1.4 --- The diversity of P2Y purinoceptor --- p.10 / Chapter 1.4.1 --- P2Y1 receptor --- p.10 / Chapter 1.4.2 --- P2Y2 receptor --- p.10 / Chapter 1.4.3 --- P2Y4 receptor --- p.10 / Chapter 1.4.4 --- P2Y6 receptor --- p.10 / Chapter 1.4.5 --- P2Y11 receptor --- p.11 / Chapter 1.5 --- The importance of calcium --- p.13 / Chapter 1.6 --- General aspects of calcium signaling --- p.14 / Chapter 1.7 --- Calcium release from the intracellular calcium stores --- p.15 / Chapter 1.7.1 --- Metabolism of inositol phosphates --- p.15 / Chapter 1.7.2 --- Ca2+ release from the internal calcium store --- p.15 / Chapter 1.8 --- Store-operated calcium channels (SOCC) or Capacitative calcium entry (CCE) --- p.18 / Chapter 1.8.1 --- The nature of the signal for CCE --- p.18 / Chapter 1.8.1.1 --- Conformational coupling --- p.18 / Chapter 1.8.1.2 --- Diffusible messenger --- p.21 / Chapter 1.9 --- Mechanism of intracellular calcium measurement --- p.25 / Chapter 1.10 --- Background of E92/3 cell line --- p.28 / Chapter Chapter 2: --- Materials and methods --- p.29 / Chapter 2.1 --- Cell culture --- p.29 / Chapter 2.2 --- Preparation of the simultaneous measurement --- p.31 / Chapter 2.2.1 --- Cell seeding --- p.31 / Chapter 2.2.2 --- Dye loading --- p.33 / Chapter 2.3 --- The setup of simultaneous measurement --- p.36 / Chapter 2.4 --- Statistical analysis --- p.40 / Chapter Chapter 3: --- Results --- p.41 / Chapter 3.1 --- Major domain of Ca2+ influx is from the basolateral side --- p.41 / Chapter 3.1.1 --- Effect of store depletion by apical ATP --- p.41 / Chapter 3.1.2 --- Effect of store depletion by basolateral ATP --- p.43 / Chapter 3.1.3 --- Effect of store depletion by thapsigargin --- p.47 / Chapter 3.2 --- Differential effect of apical and basolateral nucleotides on [Ca2+]i and Isc --- p.51 / Chapter 3.2.1 --- Basolateral ATP activates an increase in [Ca2+]i but not Isc --- p.51 / Chapter 3.2.2 --- Apical and basolateral ATP activated distinct but partially overlapped internal Ca2+ pool --- p.51 / Chapter 3.2.3 --- "Dose-dependent effect of apical or basolateral ATP, UDP and UTP on [Ca2+]i i and Isc" --- p.54 / Chapter 3.3 --- P2Y receptors subtypes on the basolateral membrane --- p.60 / Chapter 3.3.1 --- "Possible involvement of P2X, P2Y1 and P2Y11 purinoceptors on the basolateral membrane" --- p.60 / Chapter 3.3.2 --- "Cross-desensitization of experiments of UTP, ATP and UDP" --- p.60 / Chapter 3.4 --- The ATP-activated Ca2+ pool and thapsigargin-activated Ca2+ pool are partially overlapped --- p.68 / Chapter 3.5 --- Anion secretion activated by Ca2+ -independent pathway --- p.74 / Chapter Chapter 4: --- Discussion --- p.76 / Chapter 4.1 --- The major membrane for the CCE is from the basolateral side --- p.76 / Chapter 4.2 --- Basolateral P2Y receptors --- p.80 / Chapter 4.3 --- Differential effects of apical and basolateral ATP --- p.82 / Chapter 4.3.1 --- Apical and basolateral ATP release Ca2+ from different pools --- p.83 / Chapter 4.3.2 --- Ca2+ -independent mechanism --- p.83 / Chapter 4.3.3 --- Other potential signaling molecules --- p.84 / Chapter Chapter 5: --- Reference --- p.86

CoordenaÃÃo de AperfeiÃoamento de NÃvel Superior / Embora se tenha muitos estudos sobre estresse salino, pouco se conhece sobre os mecanismos individuais de sÃdio (Na+) e cloro (Cl-) na fisiologia de plantas expostas à salinidade. Baseado nisso, objetivou-se, atravÃs deste estudo, avaliar a extensÃo com que os Ãons Na+ e Cl- sÃo acumulados em diferentes partes jovens de feijÃo-caupi [Vigna unguiculata (L.) Walp.] e as suas possÃveis interferÃncias sobre as medidas de transpiraÃÃo. Para comprovar a hipÃtese que os Ãons Na+ e Cl- apresentam modelos diferentes de distribuiÃÃo nas partes vegetativas de feijÃo-caupi e que esse processo nÃo à afetado pela temperatura, foi organizada uma seqÃÃncia metodolÃgica envolvendo trÃs experimentos, onde o experimento I consistiu em avaliar a distribuiÃÃo de Na+ e Cl- em partes jovens de feijÃo-caupi submetido a concentraÃÃes crescentes de NaCl (0; 25; 50; 75 e 100 mM) durante 3 dias, onde ocorreu um maior acÃmulo de cloreto em todas as partes estudadas com exceÃÃo do caule que teve acÃmulo similar de ambos os Ãons. Baseado nessa distribuiÃÃo, utilizou-se a concentraÃÃo de 50mM, pois essa concentraÃÃo nÃo superaria a capacidade de armazenamento e, assim, seria possÃvel, nesse nÃvel, uma recuperaÃÃo (foi mantida durante 6 dias em NaCl e, a partir do 4 dia, as amostras foram divididas em dois lotes). Um lote permanecia com NaCl e o outro trocava a soluÃÃo sem NaCl e assim verificar se essa estratÃgia a planta seria capaz de recuperase. Os resultados corroboram com os obtidos no experimento I com exceÃÃo da primeira folha trifoliolada que acumulou mais Na que Cl. A literatura relata, abundantemente, que nas Ãpocas de verÃo (perÃodo do experimento I), principalmente no semi-Ãrido, as plantas acumulam mais Ãons, onde a temperatura à uma medida de fator climÃtico que pode interferir na distribuiÃÃo de Ãons e esse efeito està ligado à transpiraÃÃo. No experimento III, foi avaliado o efeito combinado de estresse salino e de temperaturas elevadas, com concentraÃÃes de 0 e 100 mM de NaCl e exposiÃÃo a temperaturas de 27; 32; 37 e 42ÂC, separadamente, por um fotoperÃodo de 12 horas. Os resultados corroboram com os obtidos, anteriormente, em que o sÃdio concentrou-se mais nas raÃzes e caules das plantas quando tratadas nas duas Ãltimas temperaturas. Em relaÃÃo ao Ãon cloreto, foi verificado um comportamento similar aos experimentos anteriores, pois este se acumulou, em maior proporÃÃo, nas folhas sob temperaturas de 32 a 42ÂC. Notadamente, o acÃmulo de cloreto està associado à quantidade de Ãgua absorvida durante a aplicaÃÃo do estresse. Em relaÃÃo à transpiraÃÃo, os resultados tambÃm corroboram com os anteriores. Dessa forma, conclui-se que, independente da concentraÃÃo, tempo, recuperaÃÃo e temperatura, o sÃdio à mais concentrado na raiz e caule, enquanto o cloreto concentra-se nas folhas. / Although it has many studies on salinity, little is known about the mechanisms of individual Na+ and Cl- in the physiology of plants exposed to salinity. Based on that meant to use this study to evaluate the extent to which the ions Na+ and Cl- are accumulated in different parts of young cowpea [Vigna unguiculata (L.) Walp.] and the possible interference on the steps of perspiration. To prove the hypothesis that the ions present different models of distribution in vegetative parts of cowpea and this process is not affected by temperature. It organized a string surrounding methodological three experiments. Where will I study was to evaluate the distribution of Na+ and Cl- young in parts of cowpea subjected to increasing concentrations of sodium chloride (0, 25, 50, 75 and 100 mM) for 3 days. In what was a greater accumulation of sodium chloride that in all parts studied, except the stem which accumulation in the equivalent proportions. Based on this distribution used with a concentration of 50 mM because it would not exceed the storage capacity and it was possible that a recovery level (it was held for 6 days in NaCl and from the 4th day the samples were divided into two lots. A lot remains NaCl and other exchanges with the solution without NaCl and thus verify that this strategy the plant would be able to recover it. The results corroborated with those obtained in the experiment I except the 1st trifoliate leaves that accumulated more in that Cl. The literature reports that plenty of times in the summer (period of the experiment I), particularly in semi-arid plants accumulate more ions. Where the temperature is a measure of climatic factors that may interfere with the distribution of ions and whether this effect is linked to transpiration. And then we performed the experiment III to assess the combined effect of salinity and high temperatures, in concentrations from 0 to 100 mM NaCl. Exposed to temperatures of 27, 32, 37 and 42 C, separately, during a photoperiod of 12 hours. The results corroborate with those obtained previously in the sodium has focused more on the roots and stems when treated in the last two temperatures. Regarding the chloride ion, was found a behavior similar to previous experiments, as they accumulated in greater proportion in the leaves at temperatures of 32 to 42 C. Notably the accumulation of chloride, is associated withthe amount of water absorbed during the implementation of stress. Regarding transpiration also corroborate the previous results. Thus it appears that regardless of concentration, time, temperature recovery and the ash is more concentrated in the root and stem while chloride is concentrated in the leaves.

INTRODUÇÃO: apesar da identificação de mais de 1500 mutações para o gene CFTR (cystic fibrosis transmembrane conductance regulator), o teste do suor ainda é o teste diagnóstico para Fibrose Cística (FC). O teste quantitativo de iontoforese por pilocarpina (TQIP) é o padrão-ouro para coleta do suor e análise do cloro, mas está sujeito a erros se não for realizado por técnicos qualificados. Embora a técnica de coleta do suor pelo sistema macroduct® e análise pela condutividade seja simples e tenha boa correlação com os níveis de cloro em estudos prévios, a mesma ainda é considerada como um teste de triagem para FC. O melhor ponto de corte para confirmar ou afastar a FC pelo método da condutividade deve ser ainda estabelecido. OBJETIVOS: comparar os valores de cloro no suor obtidos pelo teste quantitativo da iontoforese pela pilocarpina (teste clássico) com os valores de condutividade do suor obtido pelo sistema de coleta por macroduct® (teste simplificado) em pacientes com e sem FC e em uma amostra aleatória de pacientes em investigação para FC. O custo e o tempo despendidos na execução de cada teste foram também analisados na fase inicial do estudo. MÉTODOS: o teste do suor, pelas duas técnicas, foi realizado simultaneamente em pacientes com e sem FC e posteriormente em pacientes em investigação diagnóstica da doença no período de fevereiro/2006 a outubro/2008. Os pontos de corte para a condutividade para excluir ou diagnosticar FC foram < 75 e ? 90 mmol/L, respectivamente, e para o teste clássico cloro ? 60 e > 60 mmol/L. Na fase inicial da pesquisa (casos com e sem FC) foram utilizadas tabelas de contingência para os cálculos de sensibilidade (S), especificidade (E), valor preditivo positivo (VPP) e negativo (VPN), além do teste exato de Fisher para avaliar a associação entre os testes e a presença ou ausência de FC. Na amostra aleatória de pacientes usou-se a curva ROC também para os cálculos de S, E, VPP e VPN e também para calcular a área sob a curva entre os testes, e, em ambas as fases da pesquisa, para avaliar sua acurácia. Os respectivos intervalos de confiança de 95% (IC95%) também foram analisados. Para avaliação da concordância entre os testes, na amostra aleatória de pacientes, utilizou-se o coeficiente de kappa e o teste de McNemar. Aplicou-se o teste de Wilcoxon para se comparar os tempos na execução de cada teste, sendo considerados significativos quando p<0,05. RESULTADOS: 52 pacientes com FC (29M/23F; 1,5 a 18,2 anos) realizaram o teste do suor pelas duas técnicas, apresentando valores medianos de cloro e condutividade no suor de 114 e 122 mmol/L, respectivamente. A condutividade foi ? 95 mmol/L em todos os pacientes, conferindo ao teste 100% de sensibilidade (IC95%: 93,1 a 100%). Cinquenta pacientes sem FC (24M/26F; 0,5 a 12,5 anos) apresentaram valores medianos de cloro e condutividade no suor de 15,5 e 30 mmol/L, respectivamente. Em todos os casos a condutividade foi < 70 mmol/L, conferindo ao teste 100% de especificidade (IC95%: 92,9 a 100%). Foram então realizados 918 testes nos pacientes em investigação para FC, mas, em 180, as amostras foram inadequadas. Dos 738 testes realizados pelas duas técnicas, em 714 pacientes se afastou a FC, encontrando-se mediana de cloro de 11 mmol/L (variação: 3 a 137 mmol/L) e de condutividade de 25 mmol/L (variação: 14 a 138 mmol/L). Foram confirmados 24 pacientes com FC, encontrando-se uma mediana de cloro de 87 mmol/L (variação: 54 a 132 mmol/L) e de condutividade de 103 mmol/L (variação: 50 a 126 mmol/L). Pela curva ROC, com valores de condutividade > 90 mmol/L, obteve-se S= 83,3%, E= 99,7%, VPP= 90,9% e VPN= 99,4% para o diagnóstico de FC. Com valores de condutividade < 75 mmol/L praticamente se pôde excluir o diagnóstico de FC (VPN=99,7%; IC95%:99,0-100%). Houve excelente concordância entre o teste clássico e o simplificado, tanto pelo valor de kappa (0,934; IC95% 0,86 a 1,009), quanto pelo teste de McNemar (p=1,0000). O tempo despendido na execução dos testes foi significativamente menor com o teste simplificado (p<0,0001) e o custo do método simplificado foi discretamente inferior. CONCLUSÕES: o teste da condutividade do suor, seja em pacientes com diagnóstico previamente conhecido (com ou sem FC) ou quando realizado aleatoriamente, mostrou resultados superponíveis ao teste clássico e foi capaz de diferenciar pacientes com e sem FC. O teste simplificado apresentou alta sensibilidade e especificidade e houve excelente concordância entre os testes. O tempo de execução foi mais rápido e o custo inferior ao teste clássico. / INTRODUCTION: despite the identification of over 1500 CFTR (cystic fibrosis transmembrane conductance regulator) gene mutations, the sweat test is still the diagnostic test for cystic fibrosis (CF). The quantitative pilocarpine iontophoresis test (QPIT) is the gold-standard method for collection of sweat and chloride analyses, but is subjected to errors if not performed by qualified technicians. Although the technique using the macroduct system for sweat collection and the conductivity analysis is simpler and has good correlation with chloride levels in previous studies, it is still considered a screening test for CF. The best cut-off point of sweat conductivity to confirm or rule out CF must yet be established. OBJECTIVES: to compare the sweat chloride values obtained by the quantitative pilocarpine iontophoresis test (classic test) with sweat conductivity analysis obtained by the macroduct (simplified test) in patients with a confirmed CF diagnosis, in patients without CF and in a random sample of patients being investigated for CF. The cost and time spent to perform each test were also analysed in the initial phase of the study. METHODS: both techniques of sweat test were simultaneously performed initially in patients with CF, afterwards in patients in whom CF had been ruled out and finally in patients referred for a sweat test between February 2006 and October 2008. The cut-off values for sweat conductivity to exclude or diagnose CF were = 90 mmol/L and for the QPIT were sweat chloride ? 60 e > 60 mmol/L, respectively. Contingency tables were used in the initial phase of the study (cases with or without CF) for calculation of sensitivity (Se), specificity (Sp), positive (PPV) and negative predictive value (NPV) and Fisher\'s exact test was used to assess the association between the tests and the presence or absence of CF. ROC curve was used in the random sample of patients also for calculation of Se, Sp, PPV and NPV and also to calculate the area under the curve between both tests in both phases of the study to assess their accuracy. The respective 95% confidence intervals (95%CI) were also analysed. Kappa coefficient and McNemar tests were used for evaluation of agreement between the tests in the random sample of patients. Wilcoxon test was used to compare the time spent to perform each test, with the significant difference set at p < 0.05. RESULTS: in 52 CF patients (29M/23F, age range 1.5 to 18.2y) the median value of sweat Cl and conductivity were 114 and 122 mmol/L, respectively. All patients had sweat conductivity values above 95 mmol/L (100% sensitivity; 95%CI: 93.1 to 100%). In 50 patients without CF (24M/26F, age range 6m to 12.5y) the median value of sweat Cl and conductivity were 15.5 and 30 mmol/L, respectively. All patients had conductivity values bellow 70 mmol/L (100% specificity; 95%CI: 92.9 to 100%). Nine hundred and eighteen tests were then performed in patients being investigated for CF but 180 had inadequate samples. Of the 738 tests performed with both techniques in 714 CF was ruled out, with median values of sweat Cl of 11 mmol/L (range: 3 to 137 mmol/L) and of conductivity of 25 mmol/L (range: 14 to 138 mmol/L). Twenty four patients had a diagnosis of CF presenting a median sweat Cl of 87 mmol/L (range: 54 to 132 mmol/L) and a median conductivity value of 103 mmol/L (range: 50 to 126 mmol/L). The ROC curve showed that with a conductivity value > 90 mmol/L sensitivity of 83.3%, specificity of 99.7%, PPV of 90.9% and NPV of 99.4% was obtained to diagnose CF. The best conductivity cut-off value to exclude CF was < 75 mmol/L (NPV=99.7%; IC95%:99.0-100%). Good agreement were observed between the tests (kappa: 0.934; IC95% 0.86 a 1.009; McNemar test: p=1.0000). The time spent to perform the tests was significantly lower with the simplified test (p<0.0001) and the cost was slightly lower with the conductivity test. CONCLUSIONS: sweat conductivity performed in patients with a known CF or non-CF diagnosis or randomly applied in subjects referred for a sweat test showed similar results as the classic test and could differentiate patients with or without CF. Conductivity test had a high sensitivity and specificity and good agreement was observed between the techniques. The time spent to perform the tests was lower with the simplified test, as well as the cost.

Secretoglobins is a newly described polypeptide family that has gained a lot of interest in human cancer and inflammation research. Although the first secretoglobin polypeptide was discovered more than 30 years ago, their physiological function is still not known. The aim of this thesis was to study the expression of secretoglobins in normal and neoplastic human cells and tissues, and to clarify their possible involvement in human cancer. We established sensitive and specific quantitative real-time RT-PCR assays for uteroglobin, lipophilins A, B, C, mammaglobin, HIN-1, and UGRP1, and developed specific antibodies for lipophilin B and mammaglobin. By using quantitative real-time RT-PCR, immunohistochemistry, Western blotting, and in situ hybridization, we studied secretoglobin expression in normal and neoplastic cells and tissues. In normal tissues, real-time RT-PCR analysis showed high expression of mammaglobin in skin. The mammaglobin expression in skin tissue was further confirmed by in situ hybridization and immunohistochemistry, and the expression was shown to be localized to the coiled gland cells of the eccrine sweat glands and the apocrine sweat glands. In addition, we showed by using Western blotting, that mammaglobin was secreted into perspiration from the eccrine sweat glands. In pituitary gland, immunohistochemical analysis showed that lipophilin B was expressed by approximately half of the cells in the anterior pituitary. By using quantitative real-time RT-PCR it was shown that both lipophilins B and C mRNA were expressed in the pituitary gland, therefore we suggested that lipophilins B and C form heterodimers in human pituitary. In neoplastic tissues, real-time RT-PCR analysis showed dysregulated secretoglobin expression in lung tumors, with down-regulation of uteroglobin and frequent up-regulation of lipophilins A, B, C, and mammaglobin. Immunohistochemical analyses showed down-regulation of mammaglobin in cylindromas versus non-neoplastic eccrine sweat glands and of lipophilin B in pituitary adenomas versus non-neoplastic anterior pituitary. The majority of investigated cell lines showed low, or most often, lack of secretoglobin expression. Nevertheless, it has been shown that mammaglobin is over-expressed in human breast carcinomas. However, ectopic over-expression of mammaglobin and/or lipophilin B had no appreciable effect on cell proliferation rates of Hs578T breast carcinoma cells in vitro. This does not exclude the possibility that secretoglobins could confer some advantage to tumor cells in vivo, but, it indicates that the reported over-expression of mammaglobin is an epiphenomenon not causally involved in breast carcinogenesis. In summary, our major findings were that mammaglobin was expressed and secreted by the sweat glands of the skin and lipophilin B was expressed by the anterior pituitary gland; and, that expression of mammaglobin and lipophilin B were down-regulated in tumors derived from the same tissues, i.e, in cylindromas and pituitary adenomas, respectively. Furthermore, ectopic over-expression of mammaglobin and lipophilin B in breast carcinoma cells had no appreciable effect on cell proliferation rates in vitro.

The cystic fibrosis transmembrane conductance regulator (CFTR), a cAMP activated chloride channel in the apical membrane of epithelial cells, is defective in patients with cystic fibrosis (CF). Research efforts are focused on chloride channel function in order to find a cure for the disease. Genistein increased chloride transport in normal and delF508-CFTR cultured airway epithelial cells without cAMP stimulation. Prior pretreatment with phenylbutyrate did not affect the rate of the genistein-stimulated chloride efflux in these cells. S-nitrosoglutathione is an endogenous bronchodilator, present in decreased amounts in the lungs of CF patients. We studied the effect of GSNO on chloride (Cl-) transport in primary nasal epithelial cells from CF patients homozygous for the delF508-CFTR mutation, as well as in two CF cell lines, using a fluorescent Cl- indicator and X-ray microanalysis. GSNO increased chloride efflux in the CF cell lines and in primary nasal epithelial cells from CF patients. This effect was partly mediated by CFTR. If the cells were exposed to GSNO in the presence of L-cysteine, Cl- transport was enhanced after 5 min, but not after 4 h. GSNO may be a candidate for pharmacological treatment of CF patients. Chloride transport properties of cultured NCL-SG3 sweat gland cells were investigated. The CFTR protein was neither functional nor expressed in these cells. Ca2+-activated chloride conductance was confirmed and the putative Ca2+-activated chloride channel (CaCC) was further characterized in term of its pharmacological sensitivity. Corticosteroids, the primary treatment for asthma, cause necrosis/apoptosis of airway epithelial cells. It was investigated whether a newer generation of drugs used in asthma, leukotriene receptor antagonists, had similar effects. Both montelukast and dexamethasone, but not beclomethasone or budesonide induced apoptosis/necrosis in superficial airway epithelial cells. Montelukast and corticosteroids also caused decreased expression of intercellular adhesion molecule -1 (ICAM-1) in epithelial but not endothelial cells.

abstract: Fingerprints have been widely used as a practical method of biometrics authentication or identification with a significant level of security. However, several spoofing methods have been used in the last few years to bypass fingerprint scanners, thus compromising data security. The most common attacks occur by the use of fake fingerprint during image capturing. Imposters can build a fake fingerprint from a latent fingerprint left on items such as glasses, doorknobs, glossy paper, etc. Current mobile fingerprint scanning technology is incapable of differentiating real from artificial fingers made from gelatin molds and other materials. In this work, the adequacy of terahertz imaging was studied as an alternative fingerprint scanning technique that will enhance biometrics security by identifying superficial skin traits. Terahertz waves (0.1 – 10 THz) are a non-ionizing radiation with significant penetration depth in several non-metallic materials. Several finger skin features, such as valley depth and sweat ducts, can possibly be imaged by employing the necessary imaging topology. As such, two imaging approaches 1) using quasi-optical components and 2) using near-field probing were investigated. The numerical study is accomplished using a commercial Finite Element Method tool (ANSYS, HFSS) and several laboratory experiments are conducted to evaluate the imaging performance of the topologies. The study has shown that terahertz waves can provide high spatial resolution images of the skin undulations (valleys and ridges) and under certain conditions identify the sweat duct pattern. / Dissertation/Thesis / Masters Thesis Electrical Engineering 2017

Os alcaloides ß-carbolinas, Harmina e Harmalina, e o potente alucinógeno N, N-dimetiltriptamina são os principais componentes da Ayahuasca, uma bebida típica, utilizada na forma de chá e amplamente empregada em rituais religiosos na América do Sul. Embora seu uso ocorra desde períodos pré-colombianos ainda são poucas as informações sobre suas possíveis aplicações clínicas, assim como seus efeitos toxicológicos. Nos últimos anos, o uso da Ayahuasca se difundiu para vários países: Estados Unidos, Alemanha, Inglaterra, França e Espanha, com isso houve o aumento do interesse em estudos científicos acerca do chá. Em decorrência do aumento no consumo do chá, é importante o desenvolvimento e aplicação de métodos analíticos validados na rotina laboratorial e a utilização de uma matriz biológica que seja de fácil coleta e possibilite um número representativo de amostras. No presente trabalho, o suor foi utilizado como uma amostra biológica não convencional, pois possui algumas vantagens em relação às amostras convencionais como, por exemplo, o sangue; possui uma coleta não invasiva, indolor e sem constrangimentos para o voluntário, fácil aquisição de um registro cumulativo da exposição de substâncias, e janela de detecção mais longa, tornando-se uma ferramenta útil, pois permite a realização de uma coleta com maior frequência e sem atrapalhar o ritual religioso do voluntário. Embora vários métodos já tenham sido publicados para determinação de diferentes substâncias utilizando o suor como matriz biológica, tais como: ecstasy, cocaína, anfetamina, codeína, MDMA e ?9-Tetrahidrocannabinol, até a presente data, não foram encontrados métodos descritos na literatura utilizando o suor como matriz biológica para identificação de DMT, HRM e HRL em voluntários usuários da bebida. O método desenvolvido apresentou linearidade de 50 a 1500 ng/adesivo e o limite de detecção encontrado foi de 15 ng/adesivo para Harmina, 15 ng/adesivo para Harmalina e 10 ng/adesivo para o DMT. O ceficiente de variação intra-ensaio variou de 3,92% a 9,06% e o inter-ensaio variou entre 3,34% a 9,85%; a exatidão variou entre 87,5% a 105%. A recuperação encontrada foi superior a 70%. / The ß carbolines alkaloids, and Harmina Harmalina, and the potent hallucinogen N, N - dimethyltryptamine are the main components of Ayahuasca, a typical drink used in tea form widely used in religious rituals in South America. Although its use occurs from preoperative - Colombians are still little information about their possible clinical applications, as well as their toxicological effects. In recent years, the use of Ayahuasca has spread to many countries: United States, Germany, England, France and Spain, it was the increased interest in scientific studies about tea. Due to the increase in the consumption of tea, it is important the development and application of validated analytical methods for routine monitoring and the use of a biological matrix that is easily collected and allow a representative number of samples. In the present work , sweat was used as a biological sample unconventional because it has some advantages as compared to conventional samples, such as blood; have a non-invasive, painless and without constraints for voluntary, easy acquisition of a cumulative record of exposure of substances, and longer detection window, making it a useful tool collects as it allows the realization of a collection more frequently and without disturbing the religious ritual of voluntary. Although several methods have been published for the determination of different substances using sweat as a biological matrix, such as ecstasy, cocaine, amphetamine, codeine, MDMA and ?9 - Tetrahidrocannabinol, to date, no methods have been described in the literature using the sweat as a biological matrix for identification of DMT, HRL and HRM in volunteer users of the drink. The developed method was linear 50-1500 ng / patch and the limit of detection was 15 ng / patch Harmina, 15 ng / patch Harmalina and 10 ng / patch for DMT. The ceficiente intra -assay variation ranged from 3.92 % to 9.06 % and inter -assay ranged from 3.34 % to 9.85%; the accuracy ranged from 87.5 % to 105 %. The recovery was found to exceed 70%.

Title of work: Electrolyte Artist: Judit Fritz Konstfack, Ädellab   ABSTRACT How much of the ocean have I filtered while in tears? Many times, while growing up on an island have I laid eyes on the enormous surrounding body of water and wondered – is the sorrow still in there? In small portions the human body and the bodies of nature exposes each other as part of their own content. Offering a glimpse into a constant material flow. Diffusing the boundaries between object and subject. A material flow that makes it impossible to tell what anything really is, because it has already evolved. My tears taste like the ocean. So does my sweat. I Find comfort in knowing I constantly take part in something that is beyond my human comprehension. That I don’t need to fully understand why I am here, because it is obvious that I, and everyone else play a part. We just don’t know the whole scene. Though a post humanistic angle I have investigated the close relation between matter, material, the human body and the bodies of nature, to find a connection point which decreases the hierarchy between object and subject. By crystallizing matter derived from human body fluids and building a machinery that portrays the process I invite the perceiver to a closeup of their part in a never-ending system. The agency of this project is to raise questions rather than finding answers, as I believe the human is in need of reevaluating their way of thinking when it comes to material and resources. I have tried to shift focus from the human perspective and self-centeredness into a set where we participate rather than being the perceiver. Alchemy was used as both a method and as an art historical reference, as the alchemical mindset is to learn through the act of making. This has been blended with modern days technology and imagination to build a body of work that balances between science and art. While being heavily based on theory and research, the body of work is a translation in which I allowed myself to freely portray how I imagine the travelling of matter is systemized. I created a working place where studies of the unseen is concentrated and visualized. Where the many sorrows and the hard work of human beings crystallize into solids, becoming nature again. Or was it ever not? And vice versa. Throughout the project, even though separated from everything that I connect to being human, the crystals made from pure matter derived from sweat and tears could not be separated from the knowledge of their origin. Even as objects, I see them as subjects. A conclusion that shed light upon the relativeness between bodies. A conclusion that brings life into what is often seen as dead, simultaneously showing the complex machinery of an emotional being.

The theoretical part of the thesis is focused on surface treatments, their evaluation from the point of view visual properties, plasma-enhanced chemical vapor deposition and analysis of thin layers prepared by plasma polymerization. The experimental part of the thesis deals with surface treatment of stainless steel AISI 304. Thin layers of pp-HMDSO (plasma-polymer) and of DLC are deposited on the steel´s surface for the purpose of preparing transparent layer, which protect steel´s surface against of making finger prints visible. As an objective method is chosen an immersion test in artificial sweat solution according to standard ČSN EN ISO 105-E04. Evaluation of the test is measured by a gloss-meter and by a colorimeter. Prepared layers are identificated by FTIR method.

In hierdie studie ondersoek ek die ek-spreker in Breyten Breytenbach se debuutbundel. Om hierdie doel te verwesenlik, het ek 'n verteenwoordigende aantal gedigte uit die drie afdelings van die bundel in die ondersoek betrek en die ek-spreker in die gedigte nagegaan. Daar bestaan verskil van mening deur die kritici oor die "ek" in hierdie bundel. In Hoofstuk 1 wys ek hierop deur kort aanhalings. F.I.J. van Rensburg en R. Schutte beskou Die ysterkoei moet sweet as ekshibisionisties; N.P. van Wyk Louw meen daar is 'n bietjie te veel "ek-Breyten-rig-heid" in hierdie verse; A.P. Grovè oordeel dat die ek in hierdie gedigte hom soms opdring aan die leser en dit kan die gevaar inhou dat die gedig as afsonderlike struktuur aangetas kan word; Andrè P. Brink en Rob. Antonissen is van mening dat dit 'n ek-bundel sonder ekkerigheid is; D.J. Opperman meen dit is poësie wat 'n voorliefde het vir selfportrettering en selfspot, maar die verse is soms gesog en ekshibisionisties. In Hoofstuk 2 gaan ek literêr-teoreties in op die ek-spreker in die gedig. Ek probeer veral - deur bekende kritici en teoretici se uitsprake te beredeneer - antwoorde vind op o.m. die volgende vrae: Watter rol speel die ek in die gedig? Is daar 'n verskil tussen die poëtiese ek en die persoonlike ek in die gedig? Dit is ou vrae in die literatuur wat op voorbeelde af al bevredigend beantwoord is deur verskillende kritici, maar Breytenbach se poësie dwing mens om jouself opnuut rekenskap hiervan te gee - soos die verskillende oordele in Hoofstuk 1 gegee, bewys. Vanaf Hoofstuk 3 ontleed ek die bundel sistematies, dit wil sê afdeling vir afdeling en in sy samehang, met besondere aandag aan die rol van die ek-spreker in die gedig. In hierdie hoofstuk konsentreer ek slegs op die eerste gedig van die bundel, "Bedreiging van die Siekes". Dit is 'n heel ongewone gedig vir Afrikaans, 'n voorstellingsgedig waarin twee markante ek-sprekers hulle verskyning maak: 'n spottende seremoniemeester wat "Die maer man met die groen trui"- "Breyten Breytenbach" - aan 'n gehoor voorstel. Laasgenoemde figuur dra dan, vol angs en vrees, 'n gedig voor, 'n gedig wat sy problematiek openbaar: die feit dat hy vasgevang sit in 'n wêreld wat hom bedreig. Hierdie wêreld word in sterk visuele beelde voorgestel. Dis 'n surrealistiese wêreld waarin die reën wat oor alles uitsak die strate in bloed verander. Die ek-Breytenbach van die gedig sien profeties sy eie onafwendbare begrafnis: hierin móét sy gehoor 'n aandeel hê, want hy beveel hulle om hom eers te wond en dan in die grond te "plant". Intussen sal hy - in die gedig uitgewys as 'n sieke - bly praat oor die saam- en naasbestaan van uiter-stes: dood en ontbinding, en "leeubekkies" en "prewelende monnike". In die slotstrofe lewer die seremoniemeester weer spottend kommentaar op die Breytenbach-van-die-gedig: hy meen hy is skadeloos en die gehoor moet hom genadig wees. Hierdie gedig kan gesien word as opdraggedig van die bundel, in besonder van die eerste afdeling, die maer man met die groen trui - 'n titel ontleen aan hierdie gedig: veral deur die besondere procédé van selfspot en die deurlopende motiewe van angs, bedreiging, dood, verrotting én groei, ensovoort, wat telkens deur die gedigte sirkuleer. Hierdie gedig is ook net 'n "voorbeeld" - in ander "voorbeelde" word hierdie motiewe sterker uitgewerk en/of gewysig. In Hoofstuk 4 bespreek ek gedigte uit genoemde eerste afdeling. Die maer man met die groen trui - duidelik 'n bepaalde masker of karikatuur van die ware Breytenbach - het in hierdie afdeling telkens 'n wisselende aard. Hy tree op as digter, malle, sieke, vreesvolle mens, eensame, reisiger, digter-kind, ensomeer. Selde kry ons in die gedig 'n direkte oop belydenis. Breytenbach skep verskillende gestaltes van Die maer man met die groen trui in hom word dikwels houdinge en gesindhede geprojekteer wat die moderne mens in die algemeen raak. Ook in die tweede afdeling, Kopatlas - waarop ek in Hoofstuk 5 ingaan - draai die gedig nog (alhoewel in mindere mate) om ek en wêreld. Ons kry nou veral verbeeldingsreise. Zen laat hom veral sterk geld. Die openingsgedig, "nirvana", waarin Gautama Boeddha sy ek opeet, werk deur in die gedigte van hierdie afdeling. Dit vind 'n mooi hoogtepunt in "Blomme vir Boeddha", waarin die ek verdwyn en een word met al die elemente van die gedig. Hierdie eenwording in die gedig van ek en wêreld geld ook vir die derde afdeling, die mond is te geheim om pyn nie te voel nie, 'n reeks liefdesverse. (Hoofstuk 6) Veral in die liefde, in die eenwording tussen man en vrou, bestaan die moontlikheid dat hiërargieë opgehef kan word. Selfs hier het die spreker 'n wisselende aard: dit wissel van 'n nederige dankbare mens oor die gawe van die liefde en die kosbaarheid van die geliefde, van die wonderlike eenwording tussen man en vrou, van die besondere spreker wat die ongenaakbare wêreld omtower - deur sy woord - tot 'n sprokieslandskap waarin hulle gelukkig woon, tot by die 'groot figuur' ('n god) van "verwoesting, die wrak", die skepper van sy geliefde en sy wêreld. In die meerderheid liefdesverse skuil die elegiese en verwordende, tog is in die liefde die wonder van metamorfose moontlik - 'n belangrike kenmerk van hierdie poësie. Dit is uit hierdie studie duidelik dat die ek-spreker in Die ysterkoei moet sweet 'n wisselende aard het. Alhoewel die spil van die gedig in die meerderheid gedigte draai om ek en wêreld, is dit oor die algemeen nie 'n ekkerige, selfopdringerige poësie nie. Dit is digwerk wat dikwels 'n korrektief van bv. selfspot op die ek bied en binne 'n Zen-sfeer trag om ek en wêreld te versoen. / ENGLISH : In this study I examine the I-speaker in Breyten Breytenbach's first volume of poetry. In order to do this a representative number of poems were selected from the three sections in this book and the role and function of the I-speaker in these were traced. Critics differ as to how the I manifests itself in these poems. In the first chapter this is pointed out by briefly quoting a few of these differences, e.g.: According to F.I.J. van Rensburg and R. Schutte Die ysterkoei moet sweet (The iron cow must sweat) is exhibisionistic; N.P. van Wyk Louw is of the opinion that the poet is somewhat too much involved with himself (" 'n bietjie te veel ek-Breyten-rig-heid"); A.P. Grové judges that the I in some of these poems enforces itself upon the reader with the danger of impairing the ontic status of the work of art; André P. Brink and Rob. Antonissen consider these poems as being poetry of the I without an egotistic tone; for D.J. Opperman this is poetry with a preference for selfportrayal and mockery of the self; however, he finds some of the poems forced and exhibisionistic. In Chapter 2 I deal theoretically with the I-speaker in the poem. By referring to and commenting on various critics I try to arrive at answers on among others the following questions: What rôle does the I fulfil in the poem? Is there a difference between the poetic I and the personal I? These questions have been satisfactorily answered in theoretical writings, but Breytenbach's poetry forces one to answer these questions anew. From Chapter 3 onwards I analysed the selected poems. The first poem "Bedreiging van die Siekes" (Threatening of the Ill) is dealt with at length. This is a most unusual poem for Afrikaans. In this introductory poem there are two conspicious I-speakers: a jeering master of ceremonies who introduces to an audience ,"Die maer man met die groen trui" (The thin man with the green sweater) - "Breyten Breytenbach ". This Breytenbach then recites a poem which is revealing of his anxiety and fear: he is entangled in a world which threatens him. This world is portrayed through strong visual images - a surrealistic world in which the streets are transformed into blood by the rain. This speaker prophetically describes his inevitable burial in which his audience shall play a part, since he demands them to wound him first and then plant him in the ground. But he - an invalid - will continue talking about the coexistence and togetherness of extremes: death and decomposition on the one hand, "leeubekkies" (snapdragons) and "prewelende monnike" (muttering monks) on the other. In the last stanza the master of ceremonies again comments jeeringly on the Breytenbach in the poem: He is harmless, have mercy on him. This poem can be read as a dedication to the remainder of poems in the volume, and in particular to those in the first section, Die maer man met die groen trui (the thin man with the green sweater) - a title taken from the poem: particularly through the precédé of selfmockery and the continual recurrence of the motives of anxiety, threat, death, decomposition and rebirth, etc. This poem is merely an "example" - in other "examples" the same motives are more thoroughly elaborated on and/or altered. In Chapter 4 I discuss certain poems taken from the first section. The thin man with the green sweater - a mask or caricature of the real Breytenbach - takes on a changing rôle: as a poet, madman, invalid, a terrified man, one who is lonely, a traveller, a poet-child and so forth. Seldom is there a direct open confession. Breytenbach creates differing portrayals of the thin man with the green sweater; he is one with fixed likes and dislikes, and attitudes which resemble modern man in general. Also in the second section Kopatlas (mind atlas) - which is dealt with in Chapter 5 - the poems revolve around the I and the world. We find for the most part imaginary journeys. Zen asserts itself strongly. In the first poem, "nirvana", Gautama Buddha devours his I. This process dominates the remainder of the poems in this section. It culminates in "Blomme vir Boeddha" (flowers for buddha) when the I disappears and is assimilated in all the elements of the poem. This unification of the I with the world also occurs in the third section, die mond is te geheim om pyn nie te voel nie (the mouth is too secret not to feel pain) - a series of love poems. (Chapter 6) In love, in the unification between man and woman lies the possibility of hierarchies being uplifted. Even here the speaker has a changing nature: it varies between being modest, thankful for the gift of love and the beloved; the wonder of man and woman becoming one; he who with the gift of the word transforms this unkind world into a fairyland in which they live happily; to the 'important speaker' (a god) in "verwoesting, die wrak" (destruction, the wreck), the creater of his beloved and his world. In most of the love poems we find the elegiac; however, in love the wonder of metamorphosis is possible - an important characteristic of this poetry. An analysis of the poems shows the I-speaker to be of a changing nature. Although the majority of the poems focus on the I and the world, in general they do not become egotistic and self-assertive. Rather it is poetry in which we find correctives towards the I and which, within Zen, aims at reconciling the I with the world. / Dissertation (MA)--University of Pretoria, 1977. / Afrikaans / unrestricted