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by Vincent, Wai-ip Law. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1998. / Includes bibliographical references (leaves 128-134). / Abstract also in Chinese. / Chapter Chapter I. --- Literature Review / Chapter I.1. --- "Structure, functions and general physiology of equine sweat gland" / Chapter I.1.1. --- Ultrastructure of equine sweat gland --- p.1 / Chapter I.1.2. --- Functions and physiology of equine sweat gland --- p.3 / Chapter I.1.3. --- Experimental studies on equine sweat gland by functional approaches --- p.4 / Chapter I.1.4. --- Hormonal and neuronal regulation of sweat secretion in equidaes --- p.5 / Chapter I.1.5. --- Possible role(s) of extracellular ATP in equine sweat gland epithelia --- p.6 / Chapter I.1.6. --- Measurement of electrogenic anion secretion by short-circuit current (Isc) technique --- p.8 / Chapter I.2. --- Classification of purinergic receptors and its existence in biological systems / Chapter I.2.1. --- Functional classification of purinergic receptors --- p.13 / Chapter I.2.2. --- Basic structure of G-protein coupled P2Y receptors --- p.17 / Chapter I.2.3. --- Physiological function and significance of purinergic receptors --- p.20 / Chapter I.3. --- Objectives of study --- p.22 / Chapter Chapter II. --- Methods and Materials / Chapter II.l. --- Culture technique of the equine epithelial cells --- p.23 / Chapter II.2. --- Conventional short-circuit current (Isc) measurement technique / Chapter II.2.1. --- Introduction --- p.25 / Chapter II.2.2. --- Preparation of permeable supports and electrodes --- p.25 / Chapter II.2.3. --- Experimental set up and measurement of Isc --- p.28 / Chapter II.2.4. --- Measurement of Isc during experiment --- p.30 / Chapter II.3. --- Measurement of intracellular free calcium ( [Ca2+ ]ii) by microspectrofluorimetry / Chapter II.3.1. --- Preparation of cells --- p.31 / Chapter II.3.2. --- The set up and procedures for experiment --- p.31 / Chapter II.4. --- Simultaneous measurement of changes in [Ca2+ ]i and Isc / Chapter II.4.1. --- Experimental set up and manipulation --- p.35 / Chapter II.4.2. --- Other preparations before experiment --- p.37 / Chapter II.5. --- Material and solutions used for experiment / Chapter II.5.1. --- Culture media and enzyme --- p.40 / Chapter II.5.2. --- Chemicals and Drugs --- p.40 / Chapter II.5.3. --- Preparation of solution for experiments --- p.42 / Chapter II.6. --- Statistical analysis --- p.44 / Chapter Chapter III. --- Results / Chapter III.l. --- Effects of nucleotides on transepithelial ion transport / Chapter III.1.1. --- Basic electrophysiological properties of cultured equine sweat gland epithelia --- p.45 / Chapter III. 1.2. --- Short-circuit current (Isc) induced by nucleotides --- p.45 / Chapter III. 1.3. --- Identification of ion species responsible for the change in Isc --- p.50 / Chapter III. 1.4. --- Effects of chloride channels blockers on the UTP-induced Isc --- p.51 / Chapter III.2. --- Signal transduction mechanisms of P2Y-nucleotide receptors / Chapter III.2.1. --- The involvement of Gi-proteins --- p.56 / Chapter III.2.2. --- Effect of BAPTA on the increases in Isc induced by nucleotides --- p.58 / Chapter III.2.3. --- Study of P2Y-receptor mediated increase in [Ca2+]i --- p.62 / Chapter III.3. --- Characterization of the P2Y subtype(s) by cross desensitization experiments / Chapter III.3.1. --- Autologous desensitization experiments --- p.70 / Chapter III.3.2. --- Classical cross desensitization experiments --- p.70 / Chapter III.3.3. --- Characterization of the ATP-insensitive P2Y-receptor --- p.80 / Chapter III.3.4. --- Interaction between ATP and bradykinin --- p.87 / Chapter III.4. --- Simultaneous measurement of [Ca2+]i and Issc / Chapter III.4.1. --- Effect ofUDP and ADP --- p.89 / Chapter III.4.2. --- Correlation of Isc and [Ca2+]i --- p.92 / Chapter III.4.3. --- Cross desensitization experiments --- p.97 / Chapter III.5. --- Evidence of a [Ca2+]i-independent Isc-component induced by nucleotides / Chapter III.5.1. --- The time course of the ΔRf and ΔISC --- p.102 / Chapter III.5.2. --- Effect of ionomycin on the ΔISC and ΔRf induced by nucleotides --- p.110 / Chapter III.5.3. --- Effect of thapsigargin on the ΔISC and ΔRf induced by nucleotides --- p.110 / Chapter III.5.4. --- Effect of thapsigargin in nominal Ca2+-free solution --- p.115 / Chapter Chapter IV. --- Discussion / Chapter IV. 1. --- Role of extracellular nucleotides in epithelial tissues --- p.119 / Chapter IV.2. --- Characterization of an ATP-insensitive P2Y-nucleotide receptor --- p.120 / Chapter IV.3. --- Expression of the novel ATP-insensitive receptor on a functionally polarized epithelia --- p.122 / Chapter IV.4. --- Involvement of a [Ca2+]i -independent Isc induced by nucleotides --- p.124 / Chapter Chapter V. --- References --- p.128

Thesis (Ph.D.)--University of Wollongong, 2007. / Typescript. Includes bibliographical references.

M.S.

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

Submitted by Rebeka Godeiro (rebeka_carvalho@hotmail.com) on 2018-05-21T11:54:00Z No. of bitstreams: 1 NÁGELA MARIA HENRIQUE MASCARENHAS - DISSERTAÇÃO ZOOTECNIA 2018.pdf: 1661338 bytes, checksum: bfa1f2a8ffad9f48af7f40dedf2de4e9 (MD5) / Made available in DSpace on 2018-05-21T11:54:00Z (GMT). No. of bitstreams: 1 NÁGELA MARIA HENRIQUE MASCARENHAS - DISSERTAÇÃO ZOOTECNIA 2018.pdf: 1661338 bytes, checksum: bfa1f2a8ffad9f48af7f40dedf2de4e9 (MD5) Previous issue date: 2018-02-23 / Objetivou-se avaliar o efeito de duas épocas do ano, sobre a adaptabilidade de caprinos e ovinos criados em sistema extensivo no semiárido paraibano, através de parâmetros fisiológicos, testes de adaptabilidade, características estruturais do pelame, taxa de sudação e estruturas do tegumento. Foram realizados dois experimentos consecutivos. No primeiro experimento utilizou-se 24 animais, sendo 12 ovinos Santa Inês e 12 caprinos Moxotó. Avaliados oito vezes durante os meses de julho a outubro de 2016. Foram mensurados: temperatura retal (TR), frequência respiratória (FR) e temperatura superficial (TS), o índice de tolerância ao calor (ITC) e o coeficiente de tolerância ao calor (CTC). O ITC revelou que os animais são adaptados as condições climáticas. O CTC das fêmeas mostrou-se mais elevados (P<0,05) que dos machos. De maneira geral, não houve diferença significativa entre as espécies de caprinos e ovinos para a temperatura retal e a frequência respiratória. A análise de variância revelou interação significativa (P<0,05) da espécie e dentro das épocas, sobre a TS. No segundo experimento foi utilizado um delineamento inteiramente casualizado, num esquema fatorial de 2x2 (2 espécies, 2 sexos) 2x2 (2 espécies e 2 sexos) com 5 repetições. Utilizou-se 20 animais, 10 ovinos Santa Inês e 10 caprinos Moxotó. Os animais foram mantidos em sistema extensivo, avaliados apenas uma vez durante o mês de novembro, da época quente do ano de 2016. As características avaliadas foram: epessura da capa de pelo (ECP), número (NP), comprimento (CP) e peso (PS) dos pelos, a densidade númerica (DN) e volumétrica (DV). A capacidade da taxa de sudação (TS) dos animais, além de estruturas do tegumento como: folículo piloso, glândulas sudoríparas (GSu) e espessura da epiderme (EEp). O sexo dos animais não influenciou (P>0,05) em nenhuma das variavéis avaliadas. com relação as caracteristicas do pelame, o CP dos caprinos foi mais elevado (P<0,05) que os ovinos, porém o PS e a DV dos ovinos foram mais elevadas (P<0,05). Os ovinos apresentaram número de GSu e uma EEp mais elevados (P<0,05) que os caprinos. A TS não apresntou diferenças estatisticas (P>0,05). Ambas espécies estudadas demonstram estar fisiologicamente bem adaptadas e com características estrurais do pelame e esturutras do tegumento favoráveis às condições climáticas do Semiárido. / The objective of this study was to evaluate the effect of two seasons of the year on the adaptability of goats and sheep raised in an extensive system in the Paraíba semi-arid region, through physiological parameters, adaptability tests, structural characteristics of the pelage, sweating rate and integument structures. Two consecutive experiments were performed. In the first experiment 24 animals were used, 12 Santa Inês sheep and 12 Moxotó goats. Evaluated eight times during the months of July to October 2016. Rectal temperature (TR), respiratory rate (FR) and surface temperature (TS), the heat tolerance index (ITC) and the heat tolerance coefficient (CTC). The ITC has revealed that the animals are adapted to the climatic conditions. Females CTC showed higher (P <0.05) than males. In general, there was no significant difference between goat and sheep species for rectal temperature and respiratory rate. The analysis of variance revealed significant interaction (P <0.05) of the species and within the seasons, on TS. In the second experiment, a completely randomized design was used in a factorial scheme of 2x2 (2 species, 2 sexes) 2x2 (2 species and 2 sexes) with 5 replicates. Twenty animals, 10 Santa Inês sheep and 10 Moxotó goats were used. The animals were kept in an extensive system, evaluated only once during the month of November, during the hot season of 2016. The characteristics evaluated were: cape layer thickness (ECP), number (NP), length (CP) and weight (PS) of the hairs, the numerical (DN) and volumetric (DV) density. The capacity of the sweating rate (TS) of the animals, as well as structures of the integument such as: hair follicle, sweat glands (GSu) and epidermis thickness (EEp). The sex of the animals did not influence (P> 0.05) in any of the evaluated variables. (P <0.05) than sheep, but PS and BV of sheep were higher (P <0.05). The sheep had a higher GSu number and a higher EPS (P <0.05) than the goats. TS had no statistical differences (P> 0.05). Both species have been shown to be physiologically well adapted and with the characteristics of the fur and sturgeons of the tegument favorable to the climatic conditions of the Semi - arid.

Orientadores: José Dirceu Ribeiro, Francisco Ubaldo Vieira Junior / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Ciências Médicas / Made available in DSpace on 2018-08-26T00:51:01Z (GMT). No. of bitstreams: 1 SouzaGomez_CarlaCristina_M.pdf: 3648408 bytes, checksum: 8fc98c01df413aea0740fb6bb16723a1 (MD5) Previous issue date: 2014 / Resumo: Introdução: O teste do suor clássico (TSC) é aceito como padrão-ouro para o diagnóstico da fibrose cística (FC). Objetivo: Comparara estimulação e peso do suor produzido, os efeitos colaterais associados ao uso das correntes, contínua pulsada (CCP) e contínua constante (CCC) e determinar o tempo ideal para a estimulação e para a coleta de suor em indivíduos com e sem FC. Método: Estudo de intervenção prospectivo de corte transversal. Experimento 1(braço direito): CCC e CCP. Tempo de estimulação (TE) de 10min e o de coleta do suor de 30min. Correntes de 0,5; 0,75; 1,0; e 1,5mA e frequências de 0; 200; 1000; e 5000Hz. Experimento 2 (braço esquerdo): Corrente de 1,0mA; TE: 5 e 10min e coleta de 15 e 30min com frequências de 0; 200; 1000; e 5000Hz. Ambos os experimentos foram testados com densidade de corrente (DC) de 0,07 a 0,21mA/cm2. Experimento 3: Avaliar a CCP e a CCC como métodos diagnósticos para a FC comparando com diagnósticos estabelecidos por estudos na biópsia retal e sequenciamento do gene CFTR(do inglês, Cystic Fibrosis Transmembrane Condutance Regulator). Resultados: Participaram do estudo48 sujeitos (79,16% do sexo feminino), com média de 29,54±8,87 anos de idade. Não houve diferença estatística entre a interação da frequência e da corrente no peso do suor (p=0,75). Houve associação do peso do suor com a frequência de estímulo (p=0,0088) e corrente utilizada para a obtenção de sudorese (p=0,0025). A produção de suor foi maior no tempo de 10min de estimulação (p=0,0023). A coleta do suor foi maior no tempo de 30min (p=0,0019). A impedância da pele não foi influenciada pelo TE e de coleta do suor (p>0,05). A frequência da corrente utilizada mostrou associação inversa com a impedância da pele (p<0,0001). A temperatura da pele mensurada antes da estimulação foi maior que a temperatura após a estimulação (p=0,0001). No experimento 3 (29 indivíduos)a CCP mostrou melhor índice kappa comparada a CCC (0.92versus 0.52, respectivamente). Conclusão: A realização do TSC tanto com CCC quanto CCP utilizando DC de 0,14 a 0,21mA/cm2 mostrou eficácia nas etapas de estimulação e coleta de suor, sem efeitos colaterais. O tempo ideal para a estimulação e para a coleta de suor foi, respectivamente, 10 e 30min / Abstract: Background: The classic sweat test (CST) is still accepted as the goldstandard method for cystic fibrosis (CF) diagnosis. Objective: To compare the production and volume of sweat, the side-effects caused by direct pulsed current (DPC) and direct constant current (DCC) and to determine the stimulation time for stimulation and sweat for collection in CF and non-CF individuals. Method: Prospective study of cross-sectional intervention. Experiment 1 (right arm): DPC and DCC. Stimulation time (ST) of 10min and sweat collection every 30min. Currents of 0.5; 0.75; 1.0; and 1.5mA and frequencies of 0; 200; 1000; and 5000Hz. Experiment 2 (left arm): current of 1mA, ST: 5 and 10min and collection at 15 and 30min interval with frequencies of 0; 200; 1000; and 5000Hz. Both experiments were tested with current density (CD) ranging from 0.07 to 0.21mA/cm2. Experiment 3: To assess CF diagnosis by DPC and DCC methods by comparison with the established by rectal biopsy diagnosis studies and sequencing of the CFTR (Cystic Fribrosis Transmembrane Condutance Regulator) gene. Results: 48 subjects (79.16% female) with mean average of 29.54 ± 8.87 years old participated in this study. There was no statistical differences between the interaction of frequency and current in sweat weight (p=0.7488). An association was found between sweat weight with the frequency of stimulation (p=0.0088) and the current used for sweating (p=0.0025). The sweat production was higher for the 10min stimulation interval (p=0.0023). The best time interval for sweat collection was 30min (p=0.0019). The skin impedance was not influenced by ST and sweat collection time (p>0.05). The frequency of the current used was inversely associated with skin impedance (p<0.0001). The skin temperature measured before the stimulation was higher than after stimulation (p=0.0001). In experiment 3 (29 subjects), the DPC showed better kappa index compared to DCC (0.92 versus 0.52, respectively). Conclusion: ST performance with both DCC and DPC using a CD of 0.14 to 0.21mA/cm2 showed efficacy in both of stimulation and sweat collection steps, without side-effects. The optimal time for stimulation and sweat collection were, respectively, 10 and 30min / Mestrado / Saude da Criança e do Adolescente / Mestra em Ciências