A Thesis Submitted to the Faculty of Medicine University of the Witwatersrand, Johannesburg for the Degree of Doctor of Philosophy, / The cell type exclusively dealt with in this thesis is the human blood neutrophil, which is also referred to in the text as polymorphonuclear leukocyte (PMN). The experimental work in this thesis has been accomplished using one immunological and a number of biochemical investigative techniques. The former is the Boyden technique (Boyden, 1962) for the quantitative assessment of leucocyte motility. / IT2018

Thesis (Ph. D.)--University of Virginia, 1998. / Includes bibliographical references (p. 105-112). Also available online through Digital Dissertations.

Thesis (Ph. D.)--University of Washington, 2006. / Vita. Includes bibliographical references (leaves 121-137).

This document only includes an excerpt of the corresponding thesis or dissertation. To request a digital scan of the full text, please contact the Ruth Lilly Medical Library's Interlibrary Loan Department (rlmlill@iu.edu).

Tsoi Sze Man. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2005. / Includes bibliographical references (leaves 82-104). / Text in English; abstracts in English and Chinese. / Tsoi Sze Man. / Chapter Chapter 1 --- Introduction --- p.1 / Chapter 1.1 --- Overview of study --- p.1 / Chapter 1.2 --- Cerebellum --- p.2 / Chapter 1.2.1 --- General Structure of cerebellum --- p.3 / Chapter 1.2.2 --- Cell types of cerebellar cortex --- p.4 / Chapter 1.2.2.1 --- Basket cells --- p.5 / Chapter 1.2.2.2 --- Stellate cells --- p.6 / Chapter 1.2.2.3 --- Purkinje cells --- p.6 / Chapter 1.2.2.4 --- Granule cells --- p.7 / Chapter 1.2.2.5 --- Golgi cells --- p.8 / Chapter 1.2.2.6 --- Unipolar brush cells --- p.9 / Chapter 1.2.2.7 --- Deep cerebellar nuclear neurons --- p.11 / Chapter 1.2.3 --- The neuronal circuitry of the cerebellum --- p.12 / Chapter 1.2.4 --- Cerebellar function --- p.14 / Chapter 1.3 --- Cyclic nucleotide-gated (CNG) channels --- p.16 / Chapter 1.3.1 --- Molecular characterization of CNG channels --- p.16 / Chapter 1.3.2 --- Functional properties of CNG channels --- p.19 / Chapter 1.3.3 --- Expression of CNG channels in brain --- p.21 / Chapter 1.3.4 --- CNG channel and neuronal plasticity --- p.23 / Chapter 1.4 --- Hyperpolarization-activated cyclic nucleotide-gated (HCN) channels --- p.26 / Chapter 1.4.1 --- Molecular characterization of HCN channels --- p.27 / Chapter 1.4.2 --- Functional properties of HCN channels and Ih current --- p.29 / Chapter 1.4.3 --- Modulation by cyclic nucleotides --- p.31 / Chapter 1.4.4 --- Expression of HCN channels in brain --- p.33 / Chapter 1.4.5 --- Physiological roles of Ih current in central nervous system --- p.35 / Chapter 1.5 --- Aims of study --- p.38 / Chapter Chapter 2 --- Material and Methods --- p.39 / Chapter 2.1 --- Immunohistochemistry Experiments --- p.39 / Chapter 2.1.1 --- Animal preparation --- p.39 / Chapter 2.1.2 --- Tissue preparation --- p.39 / Chapter 2.1.3 --- Primary and secondary antibodies --- p.40 / Chapter 2.1.4 --- Immunofluroescence staining --- p.41 / Chapter 2.1.5 --- Confocal laser scanning microscopy and data processing --- p.41 / Chapter 2.2 --- Whole cell patch clamp recordings --- p.42 / Chapter 2.2.1 --- Brain slice preparation and identification of the cerebellar Purkinje neurons --- p.42 / Chapter 2.2.2 --- Whole cell voltage- and current-clamp recordings --- p.43 / Chapter 2.2.3 --- Drug solutions and delivery --- p.44 / Chapter 2.2.4 --- Statistical analysis --- p.45 / Chapter Chapter 3 --- Expression of Various Cyclic Nucleotide-Gated (CNG) Channel Subunits in Rat Cerebellum --- p.46 / Chapter 3.1 --- Introduction --- p.46 / Chapter 3.2 --- Results --- p.46 / Chapter 3.2.1 --- Immunoreactivity of CNGA1 in cerebellum --- p.46 / Chapter 3.2.2 --- Immunoreactivity of CNGA2 in cerebellum --- p.47 / Chapter 3.2.3 --- Immunoreactivity of CNGA3 in cerebellum --- p.47 / Chapter 3.3 --- Discussion --- p.48 / Chapter Chapter 4 --- Expression of Various Hyperpolarization-Activated Cyclic Nucleotide-Gated (HCN) Channel Subunits in Rat Cerebellum --- p.53 / Chapter 4.1 --- Introduction --- p.53 / Chapter 4.2 --- Results --- p.53 / Chapter 4.2.1 --- Immunoreactivity of HCN 1 in cerebellum --- p.53 / Chapter 4.2.2 --- Immunoreactivity of HCN2 in cerebellum --- p.55 / Chapter 4.2.3 --- Immunoreactivity of HCN3 in cerebellum --- p.55 / Chapter 4.2.4 --- Immunoreactivity of HCN4 in cerebellum --- p.55 / Chapter 4.3 --- Discussion --- p.55 / Chapter Chapter 5 --- Electrophysiological Recordings of Cyclic Nucleotide-Gated Ionic Conductance in Rat Cerebellar Purkinje Neurons --- p.59 / Chapter 5.1 --- Introduction --- p.59 / Chapter 5.2 --- Results --- p.59 / Chapter 5.2.1 --- Effect of cyclic nucleotides on the membrane potential of cerebellar Purkinje neurons --- p.59 / Chapter 5.2.2 --- Ionic conductance of the cyclic nucleotide-induced inward current --- p.61 / Chapter 5.2.3 --- The mechanism of the cyclic nucleotide-induced inward current --- p.61 / Chapter 5.2.3.1 --- Site of action --- p.62 / Chapter 5.2.3.2 --- Involvement of CNG channels and HCN channels --- p.63 / Chapter 5.2.3.3 --- Involvement of protein kinase A (PKA) and protein kinase G (PKG) --- p.65 / Chapter 5.2.3.4 --- Involvement of inwardly rectifying potassium (Kir) channels and transient receptor potential (TRP) channels --- p.65 / Chapter 5.2.4 --- Effect of cyclic nucleotides on Ih current in Purkinje neurons --- p.67 / Chapter 5.3 --- Discussion --- p.68 / Chapter Chapter 6 --- Concluding remarks References --- p.78 / References --- p.82

Thesis (Ph. D.)--University of Washington, 1998. / Vita. Includes bibliographical references (leaves [114]-128).

Thesis (Ph. D.)--University of Washington, 1999. / Vita. Includes bibliographical references (leaves 120-138).

cAMP-elevating agents such as adenosine and epinephrine (after binding to beta-adrenergic receptor) contribute to local vascular dilation and some of these dilations are endothelium-dependent. Previous intracellular Ca 2+ imaging studies in mouse microvessel endothelial cells reported that addition of adenosine or epinephrine induced a Ca2+ influx which is blocked by CNG channel blockers such as L-cis-diltiazem or LY83583. Inside-out patch clamp studies confirmed the existence of a cAMP-activated current in endothelial cells, strongly suggesting a functional role of CNG, in particular CNGA2, channels in endothelial cells. The current study went further to show that similar Ca2+ influx in response to adenosine or epinephrine occurred in endothelial cells in freshly isolated mouse aortic strips and was again blocked by L-cis-diltiazem. By measuring the isometric force developed in mouse aortic strips, we showed that CNGA2 channel-mediated Ca2+ influx in endothelial cells contributed to the endothelium-dependent vascular dilatation in response to adenosine and epinephrine. / In conclusion, cyclic nucleotides playa vital role in the regulation of intracellular Ca2+ concentration in vascular cells and Jurket T cells. / In Jurkat T cells, cyclic nucleotides regulated Ca2+ mobilization in a different way. Fluorescence-imaging studies showed that cGMP inhibited store-operated Ca2+ influx and histamine-induced Ca 2+ rise in Jurkat T cells through activation of PKG. / Thromboxane A2 (TxA2)-induced smooth muscle contraction has been implicated in cardiovascular, renal and respiratory diseases. This contraction can partly be attributed to TxA2-induced Ca2+ influx, which activates the Ca2+-calmodulin-MLCK pathway. This study aims to identify the channels that mediate TxA2-induced Ca2+ influx in vascular smooth muscle cells. Application of U-46619, a thromboxane A2 mimic, resulted in a constriction in endothelium-denuded small mesenteric artery segments. The constriction relied on the presence of extracellular Ca2+, because removal of extracellular Ca2+ abolished the constriction. This constriction was partially inhibited by a L-type Ca2+ channel inhibitor nifedipine (0.5-1 muM). The remaining component was inhibited by L-cis-diltiazem, a selective inhibitor for CNG channels, in a dose-dependent manner, Another CNG channel blocker LY83583 [6-(phenylamino)-5,8-quinolinedione] had similar effect. In primary cultured smooth muscle cells derived from rat aorta, application of U46619 (100 nM) induced a rise in cytosolic Ca2+, which was inhibited by L-cis-diltiazem. Immunoblot experiments confirmed the presence Of CNGA2 protein in vascular smooth muscle cells, These data suggest a functional role of CNG channels in U-46619-induced Ca 2+ influx and contraction of smooth muscle cells. / Leung, Yuk Ki. / "August 2010." / Adviser: Yao Xiaoxiang. / Source: Dissertation Abstracts International, Volume: 73-04, Section: B, page: . / Thesis (Ph.D.)--Chinese University of Hong Kong, 2011. / Includes bibliographical references (leaves 116-132). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Electronic reproduction. [Ann Arbor, MI] : ProQuest Information and Learning, [201-] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstract also in Chinese.

Des évidences expérimentales récentes indiquent que les ARN changent de structures au fil du temps, parfois très rapidement, et que ces changements sont nécessaires à leurs activités biochimiques. La structure de ces ARN est donc dynamique. Ces mêmes évidences notent également que les structures clés impliquées sont prédites par le logiciel de prédiction de structure secondaire MC-Fold. En comparant les prédictions de structures du logiciel MC-Fold, nous avons constaté un lien clair entre les structures presque optimales (en termes de stabilité prédites par ce logiciel) et les variations d’activités biochimiques conséquentes à des changements ponctuels dans la séquence. Nous avons comparé les séquences d’ARN du point de vue de leurs structures dynamiques afin d’investiguer la similarité de leurs fonctions biologiques. Ceci a nécessité une accélération notable du logiciel MC-Fold. L’approche algorithmique est décrite au chapitre 1. Au chapitre 2 nous classons les impacts de légères variations de séquences des microARN sur la fonction naturelle de ceux-ci. Au chapitre 3 nous identifions des fenêtres dans de longs ARN dont les structures dynamiques occupent possiblement des rôles dans les désordres du spectre autistique et dans la polarisation des œufs de certains batraciens (Xenopus spp.). / Recent experimental evidence indicates that RNA structure changes, sometimes very rapidly and that these changes are both required for biochemical activity and captured by the secondary structure prediction software MC-Fold. RNA structure is thus dynamic. We compared RNA sequences from the point of view of their structural dynamics so as to investigate how similar their biochemical activities were by computing a signature from the output of the structure prediction software MC-Fold. This required us to accelerate considerably the software MC-Fold. The algorithmic approach to this acceleration is described in chapter 1. In chapter 2, point mutations that disrupt the biochemical activity of microRNA are explained in terms of changes in RNA dynamics. Finally, in chapter 3 we identify dynamic structure windows in long RNA with potentially significant roles in autism spectrum disorders and separately in Xenopus ssp. (species of frogs) egg polarisation.