Thesis (doctoral)--Lund University, 1997. / Added t.p. with thesis statement inserted.

Thesis (doctoral)--Lund University, 1997. / Added t.p. with thesis statement inserted.

No description available.

1. The influence of catechol-O-methyltransferase inhibitor U-0521 on isotonic contraction of isolated rat vas deferens was examined to determine optimal concentration and nonspecific effects. 2. Maximum responses to (-)-epinephrine were increased at 0.4 μM and 1 μM concentrations of U-0521. Epinephrine responses were progressively decreased in the presence of higher concentrations (10 μM, 30 μM and 100 μM) Of U-0521. 3. The response to the nonadrenergic agonist neurokinin A was similarly depressed in the presence of 100 μM U-0521. 4. U-0521 not only inhibits COMT, at concentrations above 1 μM it nonspecifically depresses contraction of the rat vas deferens by both adrenergic and nonadrenergic agonists.

by Chi-shing Sum. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1995. / Includes bibliographical references (leaves 84-96). / ACKNOWLEDGEMENT --- p.i / ABSTRACT --- p.ii / LIST OF ABBREVIATIONS --- p.iv / TABLE OF CONTENTS --- p.v / Chapter CHAPTER 1 --- p.1 / Chapter 1.1 --- Biochemistry of Renin-Angiotensin System --- p.1 / Chapter 1.2 --- Physiological Roles of Angiotensin --- p.5 / Chapter 1.3 --- Biochemistry of Angiotensin Receptors --- p.6 / Chapter 1.4 --- Tissue Renin-Angiotensin System --- p.13 / Kidney --- p.13 / Blood vessels --- p.14 / Heart --- p.15 / Brain --- p.16 / Testes --- p.17 / Chapter 1.5 --- Structure and Function of Vas Deferens --- p.18 / Chapter 1.6 --- Aim of Study --- p.21 / Chapter CHAPTER 2 --- p.22 / Chapter 2.1 --- introduction --- p.22 / Chapter 2.2 --- Materials --- p.23 / Chapter 2.3 --- methods --- p.23 / Chapter 2.3.1 --- Preparation of isolated epididymal rat vas deferens --- p.23 / Chapter 2.3.2 --- Concentration-responses to angiotensins --- p.24 / Chapter 2.3.3 --- Effects of angiotensin II in the presence of protease inhibitors --- p.24 / Chapter 2.3.4 --- Effect of losartan and CGP 42112 --- p.24 / Chapter 2.3.5 --- Schild analysis --- p.25 / Chapter 2.3.6 --- Interaction of angiotensin II with exogenous noradrenaline --- p.25 / Chapter 2.3.7 --- Statistical analysis --- p.25 / Chapter 2.4 --- results --- p.25 / Chapter 2.4.1 --- Effect of angiotensin on epididymal rat vas deferens --- p.25 / Chapter 2.4.2 --- Concentration-responses to angiotensins in epididymal rat vas deferens --- p.27 / Chapter 2.4.3 --- Effect of angiotensin II in the presence of protease inhibitors --- p.27 / Chapter 2.4.4 --- Effect of losartan ami CGP 42112 --- p.27 / Chapter 2.4.5 --- Schild analysis --- p.36 / Chapter 2.4.6 --- Interaction of angiotensin II with exogenous noradrenaline --- p.36 / Chapter 2.5 --- Discussion --- p.36 / Chapter CHAPTER 3 --- p.39 / Chapter 3.1 --- Introduction --- p.39 / Chapter 3.2 --- Materials --- p.39 / Chapter 3.3 --- Methods --- p.40 / Chapter 3.3.1 --- Preparation of isolated prostatic rat vas deferens --- p.40 / Chapter 3.3.2 --- Concentration-responses to angiotensins --- p.40 / Chapter 3.3.3 --- Effects of angiotensin II in the presence of protease inhibitors --- p.41 / Chapter 3.3.4 --- Effect oflosartan and CGP 42112 --- p.41 / Chapter 3.3.5 --- Schild analysis --- p.41 / Chapter 3.3.6 --- Interaction of angiotensin II with exogenous noradrenaline --- p.42 / Chapter 3.3.7 --- Concentration-response to angiotensin II after reserpine treatment --- p.42 / Chapter 3.3.8 --- Concentration-response to angiotensin II after desensitization of P2-purinoceptors --- p.42 / Chapter 3.3.9 --- Statistical analysis --- p.42 / Chapter 3.4 --- Results --- p.43 / Chapter 3.4.1 --- Effect of angiotensin on prostatic rat vas deferens --- p.43 / Chapter 3.4.2 --- Concentration-responses to angiotensins in prostatic rat vas deferens --- p.43 / Chapter 3.4.3 --- Effect of angiotensin II in the presence of protease inhibitors --- p.43 / Chapter 3.4.4 --- Effect of losartan and CGP 42112 --- p.49 / Chapter 3.4.5 --- Schild analysis --- p.49 / Chapter 3.4.6 --- Interaction of angiotensin II with exogenous noradrenaline --- p.49 / Chapter 3.4.7 --- Concentration-response to angiotensin II afier reserpine treatment --- p.54 / Chapter 3.4.8 --- Concentration-response to angiotensin II after desensitization of P2-purinoceptors --- p.54 / Chapter 3.5 --- Discussion --- p.63 / Chapter CHAPTER 4 --- p.66 / Chapter 4.1 --- introduction --- p.66 / Chapter 4.2 --- Materials and Methods --- p.66 / Chapter 4.2.1 --- Preparation of polyclonal angiotensin II antiserum --- p.66 / Chapter 4.2.1.1 --- Preparation of peptide conjugate --- p.66 / Chapter 4.2.1.2 --- Protein determination --- p.67 / Chapter 4.2.1.3 --- Immunization of rabbit with peptide conjugate --- p.67 / Chapter 4.2.1.4 --- Collecting rabbit serum --- p.68 / Chapter 4.2.2 --- Characterization of BSA-Ang II and Thy-Ang II antisera --- p.68 / Chapter 4.2.2.1 --- Slot blotting --- p.68 / Chapter 4.2.2.2 --- Enzyme-linked immunosorbent assay (ELISA) --- p.69 / Chapter 4.3 --- RESULT --- p.69 / Chapter 4.3.1 --- Preparation of polyclonal angiotensin II antiserum --- p.69 / Chapter 4.3.2 --- Characterization of BSA-Ang II and Thy-Ang II antisera --- p.70 / Chapter 4.3.2.1 --- Slot blotting --- p.70 / Chapter 4.3.2.2 --- Enzyme-linked immunosorbent assay (ELISA) --- p.70 / Chapter 4.3 --- discussion --- p.76 / Chapter CHAPTER 5 --- p.78 / Chapter 5. 1 --- General Discussions --- p.78 / REFERENCES --- p.84 / APPENDIX --- p.97 / Published Abstract and Paper --- p.97

Relapse to drug taking is a major factor contributing to the low success rate of opioid addiction treatment programmes. Recently, studies have revealed a buprenorphine/naltrexone combination had successfully increased the treatment retention rate (compared to naltrexone alone) among heroin addicts (with history of cocaine abuse) who had undergone detoxification. However, buprenorphine and naltrexone could not be administered as a single formulation due to their different bioavailability, which could create compliance issues. Therefore, in this project, we aimed to synthesise a series of ligands each having the pharmacological profile of the buprenorphine/naltrexone combination (partial agonist (ORL-1 receptors), antagonist (u- and x-opioid receptors)). Based on the group's previous work, this profile can be achieved within the orvinols series. Compound BU127, a buprenorphine analogue with phenyl substituent (C20) is very close to the desired profile. Therefore, in order to optimize BU127's profile, we designed and synthesised a series of aromatic analogues, including analogues with a small group attached to the aromatic system to increase the ORL-1 receptor efficacy, while retaining the low efficacy / antagonist activity at the u-opioid receptor and antagonist activity at x-opioid receptor. However, [35S]GTPyS screening has shown a sudden increase of x-opioid receptor efficacy with these modifications. The related compound BU10119, having a Cv-methyl, met the desired profile at all targeted receptors in the [35S]GTPyS screen. A few analogues were selected for further evaluation in functional assays in the isolated tissue preparations (rat vas deferens (for the ORL-1 and u-opioid receptors) and mouse vas deferens (for the K-opioid receptor)) to estimate their binding affinity (Ks) and potency (pA2) of the compounds relative to buprenorphine, using Schild analysis and Schild equation. Of the analogues synthesised, only compounds BU127 and BU1 0119 have met the desired profile at the targeted receptors (competitive reversible at the ORL-1 and u-opioid receptors) and having binding affinity at each receptor similar to buprenorphine (ORL-1, ~- and K-opioid receptors). Based on these results, at this point, the optimum features of buprenorphine analogues in order to achieve the targeted profiles are having a small group at Cy and a 6-membered aromatic substituent at C . 20 Without any substituent group attached to the aromatic ring.

No description available.

Cocaine- and amphetamine-regulated transcript (CART) is a novel family of peptides, of which CART peptide fragments 55-102 and 62-102 are reported to be the endogenous, physiologically active peptides. Immunohistochemical studies with an antiserum directed against the CART peptide fragment 55-102 revealed CART-like immunoreactive (CART-LI) nerve fibers in the rat epididymis. The number was highest in the cauda epididymis and became progressively fewer toward the caput epididymis; the vas deferens exhibited an abundance of CART-LI fibers. Injection of the retrograde tracer Fluorogold (Fluorochrome, Inc., Englewood, CO) to the junction between the vas deferens and cauda epididymis labeled a large number of neurons in the major pelvic ganglion, some of which were CART-positive. Double-labeling the ganglion sections with tyrosine hydroxylase (TH) and CART antisera revealed that CART-LI and TH-LI were expressed in two distinct populations of ganglion cells. Some of the TH-LI cells in the ganglia, however, were covered with web-like CART-LI endings. The effects of CART peptide 55-102, referred to herein as CART, on anion secretion in the form of short circuit currents (Isc) were assessed in cultured epithelia. The CART (1 to 5 μM) applied to the basolateral or apical side of the cultured epithelia caused no significant responses on Isc, whereas lys-bradykinin (1 μM) produced a large Isc response in the same preparations. Our results show that CART-LI is present in a population of rat pelvic ganglion cells, which may give rise to CART-LI nerve fibers as observed in the vas deferens and the epididymis. The biological function of CART in the rat epididymis is not known, but it apparently is not involved in ion secretion across the epithelium.

Doctor of Philosophy / Department of Anatomy and Physiology / Bruce Schultz / Transforming growth factor β1 (TGF-β1) is a cytokine that reportedly affects the severity of cystic fibrosis lung disease. The goal of this project was to define the effect of TGF-β1 on vas deferens, an organ that is universally affected in male cystic fibrosis patients. In the first study, experiments were conducted using freshly isolated porcine vas deferens epithelial cells. Primary porcine vas deferens epithelial cells exposed to TGF-β1 exhibited a significantly reduced basal transepithelial electrical resistance (Rte). TGF-β1-induced reduction in Rte was prevented by SB431542, a TGF-β receptor I inhibitor, indicating that the effect of TGF-β1 requires the activation of TGF-β receptor I. Western blot and immunohistochemistry results showed the expression of TGF-β receptor I in native vas deferens epithelia, indicating that the impaired barrier function and anion secretion that were observed in cultured vas deferens cells can likely be observed in the native context. Immunohistochemical outcomes showed that TGF-β1 exposure led to loss of organization of tight junction proteins occludin and claudin-7. These outcomes suggest that TGF-β1 impairs the barrier integrity of epithelial cells lining the vas deferens. In a parallel study that employed PVD9902 cells that are derived from porcine vas deferens, TGF-β1 exposure significantly reduced anion secretion stimulated by forskolin, forskolin/IBMX, and 8-pCPT-cAMP, suggesting that TGF-β1 affects downstream targets of the cAMP signaling pathway. Real-time RT-PCR and western blot analysis showed that TGF-β1 exposure reduced both the mRNA and the protein abundance of cystic fibrosis transmembrane conductance regulator (CFTR). Pharmacological studies showed that the inhibitory effect of TGF-β1 on forskolin-stimulated anion secretion was abrogated by SB431542 and attenuated by SB203580, a p38 mitogen-activated protein kinase (MAPK) inhibitor. These outcomes suggest that TGF-β1, via the activation of TGF-β receptor I and p38 MAPK signaling, reduces CFTR expression, and thus impairs CFTR-mediated anion secretion. Outcomes from these studies suggest that, in epithelial cells lining the vas deferens, TGF-β1 exposure leads to an impaired physical barrier and/or reduced anion secretion, which is expected to modify the composition and the maintenance of the luminal environment and thus, is expected to reduce male fertility.

Características morfológicas, histomorfométricas, ultraestruturais e tridimensionais do ducto deferente de codorna doméstica (Coturnix coturnix) da variedade italiana foram estudadas e descritas. Para tanto, foram utilizadas 30 codornas sexualmente maduras, procedentes de um criatório da Fazenda Edgardia de Botucatu, Estado de São Paulo, Brasil. Os ductos deferentes foram coletados, observados e documentados ao longo do ano nas estações de outono, inverno, primavera e verão, e estudados por métodos de macro-mesoscopia, microscopia de luz, microscopias eletrônicas de transmissão (MET) e de varredura (MEV). Cada ducto deferente foi reduzido em três segmentos: proximal ou adepididimal; médio ou justa-renal, e distal ou adcloacal. Os estudos ultraestruturais e tridimensionais se concentraram no segmento médio do ducto. Diferenças marcantes na morfologia foram observadas, incluindo características estruturais e histomorfométricas, sobre os valores médios de altura epitelial, diâmetros tubular e luminal dos ductos deferentes, mensurados durante as quatro estações do ano, sendo mais marcantes as observações feitas no outono, a fase quiescente, e na primavera, a fase ativa, do ciclo testicular anual. Nas fases intermediárias do ciclo, compreendendo os meses de inverno (fase recrudescente), e de verão (fase regressiva), os reflexos da cinética testicular sobre a estrutura do ducto deferente se fizeram notar principalmente nas características subcelulares das células principais do epitélio de revestimento. As observações de MEV confirmaram a variabilidade morfológica do ducto deferente nas estações marcantes do ano (outono e primavera) em termos das fases do ciclo testicular anual. Estes estudos permitiram concluir que esta espécie de ave mostra um padrão circanual no ciclo reprodutivo, influenciando as características morfológicas dos ductos excretores do testículo / The morphological, histomorphometric, ultrastructural and three-dimensional features of the vas deferens of quail (Coturnix coturnix) of the Italian variety were studied and described. For this study were utilized 30 adult sexually active quails taken from the experimental Edgardia Farm of the UNESP, Botucatu, São Paulo, Brazil. The vas deferens was collected, observed and documented along the year in the autumn, winter, spring and summer seasons, and studied by macro-mesoscopy, light microscopy, transmission electron microscopy (TEM) and scanning electron microscopy (SEM). Each vas deferens was reduced in three segments: proximal (adepididymal); middle (just the median lobe of the kidney); and distal (adcloacal). The ultrastructural and three-dimensional studies were concentred in the middle segment of the duct. It was verified marked morphologic differences in the vas deferens, including structural and histomorphometric features concerning the middle values of the epithelial height, tubular and luminal diameters measured during the four seasons of the year. The observations made in the autumn the quiescent phase and in the spring the active phase of the annual testicular cycle were more evident. In the intermediaries phases of the cycle, i.e., the winter (recrudescent phase) and summer (regressive phase), the influence of the testicular kinetic on the vas deferens structure was noted mainly in the subcellular characteristics of principal cells of the epithelium lining. The SEM observations confirmed the morphologic variability of the vas deferens in the main seasons of the year (autumn and spring) concerning to the annual testicular cycle phases. These studies allowed to conclude that this avian species showed a circunannual pattern of the reproductive cycle, influencing the morphological features of the excurrent ducts of the testis