Indiana University-Purdue University Indianapolis (IUPUI) / The aim of this current study was to determine if the exposure of human dental pulp stem cells (HDPSC) to the DynaMatrix membrane will result in an increased production of angiogenic cytokines that are critical for pulp/root regeneration. Angiogenesis cytokine arrays have been established as a viable method for assessing expression of cytokines.20 HDPSC were chosen as they are expected to be found in the apical papilla and the infected immature root canal system of teeth that current regenerative endodontic techniques are designed to treat.

Cai, Ganhui. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2013. / Includes bibliographical references (leaves 160-192). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstract also in Chinese.

Chronic treatment with enalapril and valsartan significantly improved endothelium-dependent relaxations of aortas from ovariectomized rats. The present results clearly point to that chronic treatment with enalapril or valsartan reduced expression and function of RAS and associated oxidative stress, thereby augmented NO bioavailability and improved endothelium-dependent relaxations. These results provided novel evidence supporting a potential application of ACEI and ARB in the treatment of endothelial dysfunction-associated vascular complications in postmenopausal women. / Functional studies showed that acetylcholine-induced relaxations in isolated aortas were impaired in a time-dependent manner, from the 4th-week to the 12th-week after ovariectomy. The impaired relaxations were partially restored by acute treatment with losartan [angiotensin II type 1 receptor (AT1R) blocker] and apocynin [NAD(P)H oxidase inhibitor]. The present results demonstrate that estrogen deficiency blunted endothelium-dependent relaxations due to impaired the NO bioavailability, which is closely associated with the reduced eNOS activity and elevated RAS expression and associated NAD(P)H oxidase-mediated oxidative stress in the vascular wall. / The present study shows that chronic consumption of cranberry juice restored the endothelium-dependent relaxations in aortas from ovariectomized rats. In ovariectomized rats, the phenylephrine-induced a higher active vascular tension; which was prevented by chronic consumption of cranberry juice. The present data also shows that cranberry juice administration significantly reduces the elevated serum levels of total cholesterol, triglyceride, high density lipoprotein (HDL) cholesterol, non-HDL (nHDL) cholesterol, and nHDL/HDL. The active ingredients in the cranberry juice organic extract accounting for the vascular benefit remain to be further examined even though the extract causes endothelial NO-dependent relaxations in normal rat aortas and contains several bioactive compounds, some of which may protect the vascular function. This study provides the first line of evidence concerning a significant vascular benefit of chronic consumption of cranberry juice during estrogen deficiency. (Abstract shortened by UMI.) / The present study used ovariectomized female rats that mimic the "equivalent" state of menopause in human and investigated whether dysregulation of RAS components contribute to endothelial dysfunction and whether chronic treatment with ACEI (enalapril) or ARB (valsartan) could restore endothelial function in ovariectomized rats. / The second objective of the present study was to investigate whether or not consumption of cranberry juice, a popular drink in Western countries, could restore endothelial function during estrogen deficiency and to elucidate the cellular mechanisms underlying the improved endothelial function. / Yung, Lai Ming. / Adviser: Huana Yu. / Source: Dissertation Abstracts International, Volume: 70-06, Section: B, page: 3252. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2008. / Includes bibliographical references (leaves 148-168). / 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, [200-] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstracts in English and Chinese. / School code: 1307.

PrPc é uma glicoproteína de 35 KDa, bastante conservada entre as espécies e essencial no processo de transmissão e patogênese de várias doenças neurodegenerativas como a encefalopatia espongiforme bovina e a doença de Creutzfeldt-Jacob (PRUSINER, 1991). Embora sua função fisiológica ainda seja desconhecida, sabe-se que a patogênese das doenças priônicas requer a sua expressão e é freqüentemente acompanhada do acúmulo no cérebro de uma isoforma anormal de PrPc, designada PrPsc (GABIZON e cols., 1997). Interessado nos possíveis papéis fisiológicos da proteína PrPc, nosso grupo tem se dedicado a estudar as interações que PrPc realiza com outras moléculas. Identificamos e caracterizamos duas interações nas quais PrPc está envolvido: com uma proteína ligante de 66 KDa, recém-identificada como sendo a proteína STI1 (ZANATA e cols., 2002) e com a principal proteína não colagênica da matriz extracelular, a laminina (GRANER e cols., 2000). No presente trabalho, procuramos investigar as vias de sinalização deflagradas por cada uma dessas interações, como também o seu papel nos mecanismos de morte celular programada e memória. Os resultados apresentados nesse trabalho demonstram que a interação PrPc-p66 desencadeia uma resposta neuroprotetora na camada neuroblástica da retina de roedores em desenvolvimento via cAMP/PKA. Além disso, verificamos que a interação PrPc-laminina desempenha um importante papel na formação da memória de curta duração através da ativação da via cAMP-PKA-MAPK, e na memória de longa duração ativando somente a via cAMP/PKA. / PrPc is an extremely conserved 35 KDa glycoprotein which seems to be essential during the transmission and pathogenesis of several neurodegenerative diseases like bovine spongiform encephalopathy or Creutzfeldt-Jacob disease (PRUSINER, 1991). Although the physiological function of this protein remains unclear, it is well established that prion diseases require PrPc expression and are often characterized by deposition of an abnormal PrPc isoform, named PrPsc (GABIZON et. al., 1997). Interested in the normal fuction of PrPc, our group has been dedicated to study the interations that PrPc could entertain with other molecules. We have identified and characterized two interactions in which PrPc is involved: with a 66 KDa ligand protein, recently identified as the STI1 protein (ZANATA et. al., 2002) and with laminin (GRANER et. al., 2000). In this work, we have investigated the signaling pathways triggered by these interactions, as well as their relevance in programmed cell death and memory formation mechanisms. We show in this work that PrPc-p66 interaction transduces neuroprotective signals through a cAMP/PKA-dependent pathway in the neuroblastic layer of rodents\' retina. Moreover, we demonstrated that PrPc-laminin interaction has an important role for short-term memory formation through the activation of cAMP-PKA-MAPK pathways and for long-term memory with the activation of the cAMP/PKA pathway only.

PrPc é uma glicoproteína de 35 KDa, bastante conservada entre as espécies e essencial no processo de transmissão e patogênese de várias doenças neurodegenerativas como a encefalopatia espongiforme bovina e a doença de Creutzfeldt-Jacob (PRUSINER, 1991). Embora sua função fisiológica ainda seja desconhecida, sabe-se que a patogênese das doenças priônicas requer a sua expressão e é freqüentemente acompanhada do acúmulo no cérebro de uma isoforma anormal de PrPc, designada PrPsc (GABIZON e cols., 1997). Interessado nos possíveis papéis fisiológicos da proteína PrPc, nosso grupo tem se dedicado a estudar as interações que PrPc realiza com outras moléculas. Identificamos e caracterizamos duas interações nas quais PrPc está envolvido: com uma proteína ligante de 66 KDa, recém-identificada como sendo a proteína STI1 (ZANATA e cols., 2002) e com a principal proteína não colagênica da matriz extracelular, a laminina (GRANER e cols., 2000). No presente trabalho, procuramos investigar as vias de sinalização deflagradas por cada uma dessas interações, como também o seu papel nos mecanismos de morte celular programada e memória. Os resultados apresentados nesse trabalho demonstram que a interação PrPc-p66 desencadeia uma resposta neuroprotetora na camada neuroblástica da retina de roedores em desenvolvimento via cAMP/PKA. Além disso, verificamos que a interação PrPc-laminina desempenha um importante papel na formação da memória de curta duração através da ativação da via cAMP-PKA-MAPK, e na memória de longa duração ativando somente a via cAMP/PKA. / PrPc is an extremely conserved 35 KDa glycoprotein which seems to be essential during the transmission and pathogenesis of several neurodegenerative diseases like bovine spongiform encephalopathy or Creutzfeldt-Jacob disease (PRUSINER, 1991). Although the physiological function of this protein remains unclear, it is well established that prion diseases require PrPc expression and are often characterized by deposition of an abnormal PrPc isoform, named PrPsc (GABIZON et. al., 1997). Interested in the normal fuction of PrPc, our group has been dedicated to study the interations that PrPc could entertain with other molecules. We have identified and characterized two interactions in which PrPc is involved: with a 66 KDa ligand protein, recently identified as the STI1 protein (ZANATA et. al., 2002) and with laminin (GRANER et. al., 2000). In this work, we have investigated the signaling pathways triggered by these interactions, as well as their relevance in programmed cell death and memory formation mechanisms. We show in this work that PrPc-p66 interaction transduces neuroprotective signals through a cAMP/PKA-dependent pathway in the neuroblastic layer of rodents\' retina. Moreover, we demonstrated that PrPc-laminin interaction has an important role for short-term memory formation through the activation of cAMP-PKA-MAPK pathways and for long-term memory with the activation of the cAMP/PKA pathway only.

Indiana University-Purdue University Indianapolis (IUPUI) / Literature has demonstrated the ability of human adipose tissue to generate large amounts of lactate. However, it is not understood why adipose tissue produces lactate, how the production of lactate is regulated, and what potential benefit this has to the adipocyte or the organism. We first characterized a human model of adipogenic differentiation with minimal donor to donor variability to assess metabolic changes associated with mature adipocytes compared to their precursors. Indeed, similar to what was observed in human clinical studies, the differentiated adipocytes demonstrated increased lactate production. However, the differentiated adipocytes compared to their precursors (preadipocytes or ASCs) demonstrate an aerobic glycolysis-like (also called Warburg effect-like) increase in glycolysis characterized by a 5.2 fold increase in lactate production in normoxic conditions (atmospheric oxygen tension). Remarkably, this increase in lactate occurred even though the differentiated adipocytes simultaneously demonstrate an increase in oxidative capacity. This low fraction of oxidative capacity coupled with increased lactate production indicated regulation of oxidative rates most likely at the point of pyruvate conversion to either acetyl-CoA (oxidative metabolism) or lactate (glycolytic metabolism). To investigate the potential regulation of this metabolic phenotype, PDK isoform expression was assessed and we found PDK 1 and 4 transcript and protein elevated in the differentiated cells. Non-selective pharmacologic inhibition of the PDKs resulted in decreased lactate production, supporting a regulatory role for PDK in modulation of the observed Warburg effect. PDK inhibition also resulted in increased ROS production in the adipocytes after several hours of treatment and a decrease in cell viability when PDK inhibition was carried out over 36 hours. The resulting loss in viability could be rescued by antioxidant (Tempol) treatment, indicating the decrease in viability was ROS mediated. Similar to what is seen in cancer cells, our data demonstrate that differentiation of human adipocytes is accompanied by a PDK-dependent increase in glycolytic metabolism (Warburg effect) that not only leads to lactate production, but also seems to protect the cells from increased and detrimental generation of ROS.

No description available.

Kattaya-Annappa-Seema. / Thesis submitted in: December 2006. / "2+" and "+" in the title are superscripts. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2007. / Includes bibliographical references (leaves 162-188). / Abstracts in English and Chinese. / Abstract --- p.i / Acknowledgements --- p.viii / Abstracts published based on work in this thesis --- p.ix / Table of contents --- p.x / Chapter Chapter 1: --- Introduction / Chapter 1.1 --- Pulmonary hypertension / Chapter 1.1.1 --- Pulmonary circulation and its functions --- p.1 / Chapter 1.1.2 --- Pulmonary vascular diseases and symptoms --- p.3 / Chapter 1.2 --- Muscarinic Receptor functions --- p.5 / Chapter 1.3 --- Acetylcholine (ACh) and its function --- p.7 / Chapter 1.4 --- ACh receptors in pulmonary vascular bed --- p.11 / Chapter 1.5 --- Potassium channel classification and functions --- p.12 / Chapter 1.5.1 --- "Importance of High-conductance, Ca2+ activated potassium channel (BKca) in vascular smooth muscle functions" --- p.15 / Chapter 1.5.2 --- Modulation of BKca channel by various cations --- p.18 / Chapter 1.6 --- Calcium signaling and homeostasis --- p.20 / Chapter 1.7 --- Role of sodium in hypertension --- p.22 / Chapter 1.8 --- Na+-H+ exchanger (NHE) functions --- p.25 / Chapter 1.9 --- Na+-Ca2+ exchanger (NCX) in vascular smooth muscle cells --- p.29 / Chapter 1.10 --- Spontaneously hypertensive rat (SHR) / Chapter 1.10.1 --- Hypertension in SHR --- p.32 / Chapter 1.10.2 --- BKca in smooth muscle vasculature of SHR --- p.33 / OBJECTIVES OF THE STUDY --- p.34 / Chapter Chapter 2: --- Material and methods / Chapter 2.1 --- Material / Chapter 2.1.1 --- Solutions and Drugs --- p.35 / Chapter 2.1.2 --- Chemicals and Enzymes --- p.39 / Chapter 2.2 --- Methods / Chapter 2.2.1 --- Isolation of single pulmonary arterial smooth muscle cells --- p.40 / Chapter 2.2.2 --- Electrophysiological measurement --- p.42 / Chapter 2.2.3 --- Data analysis --- p.44 / Chapter Chapter 3: --- Receptor-mediated activation of BKca Channel / Chapter 3.1 --- BKCa activation by ACh/ Carbachol (CCh) --- p.45 / Chapter 3.2 --- Role of extracellular sodium ([Na+]o）on BKca activation --- p.49 / Chapter 3.3 --- Receptor-mediated activation of BKca in a [Na+]o-containing solution --- p.51 / Chapter 3.4 --- Receptor-mediated activation of BKca in a [Na+]o-free solution --- p.55 / Chapter Chapter 4: --- Non-receptor mediated activation of BKCa Channel / Chapter 4.1 --- Effect of different concentrations of sodium nitroprusside (SNP) on BKCa activation --- p.60 / Chapter 4.2 --- Effect of SNP on BKca activation in a [Na+]o-containing and [Na+]o-free solutions --- p.62 / Chapter Chapter 5: --- Role of NHE in modulating activation of BKCa Channel / Chapter 5.1 --- Effect of Monensin on BKca activation / Chapter 5.1.1 --- Effect of monensin on CCh-mediated activation of BKca in a [Na+]o-containing solution --- p.70 / Chapter 5.1.2 --- Effect of monensin on CCh-mediated activation of BKca in a [Na+]o-free solution --- p.74 / Chapter 5.1.3 --- Effect of monensin on SNP- mediated activation of BKca in [Na+]o-containing and [Na+]o-free solutions --- p.78 / Chapter 5.2 --- Effect of 5-(N-ethyl-N-isopropyI) amiloride (EIPA) on BKCa activation / Chapter 5.2.1 --- Effect of EIPA on CCh-mediated activation of BKca in a [Na+]o-containing solution --- p.85 / Chapter 5.2.2 --- Effect of EIPA on CCh-mediated activation of BKca in a [Na+]。-free solution --- p.89 / Chapter 5.2.3 --- Effect of EIPA on SNP-mediated activation of BKCa in [Na+]o-containing and [Na+]o-free solutions --- p.93 / Chapter Chapter 6: --- Role of NCX in modulating activation of BKCa Channel / Chapter 6.1 --- Effect of KB-R7943 on CCh-mediated activation of BKCa in a [Na+]o-containing solution --- p.100 / Chapter 6.2 --- Effect of KB-R7943 on CCh-mediated activation of BKCa in a [Na+]o-free solution --- p.104 / Chapter 6.3 --- Effect of KB-R7943 on SNP-mediated activation of BKca in [Na+]o-containing and [Na+]o-free solutions --- p.109 / Chapter Chapter 7: --- Effect of intracellular sodium ([Na+]i) on BKCa channel activation / Chapter 7.1 --- Effect of CCh on BKCa channel activation with elevated [Na+]i pipette solution --- p.117 / Chapter 7.2 --- Effect of SNP on BKca channel activation with elevated [Na+]j pipette solution --- p.130 / Chapter Chapter 8: --- Discussion / Chapter 8.1 --- Modulatory effect of ACh and SNP --- p.138 / Chapter 8.2 --- Role of ion exchangers: NHE and NCX in modulating BKca channel function --- p.144 / Chapter 8.3 --- Modulatory effect of elevated [Na+]i on BKca activation --- p.153 / CONCLUSION --- p.161 / References --- p.162

Células mononucleares de sangue de cordão umbilical (SCU) e sangue periférico mobilizado (SPM) com G-CSF, foram cultivadas in vitro com citocinas, na presença ou não de estroma de medula óssea. Os objetivos foram avaliar a capacidade proliferativa de células progenitoras, a ocorrência de apoptose e expressão de integrina. Nas culturas sem estroma, a celularidade aumentou 5 vezes (SCU) e não se alterou nas de SPM. O total de células CD34+ caiu em ambas culturas. Com estroma, o total de células nucleadas aumentou 7 vezes (SCU) e 2,3 vezes (SPM). O total de células CD34+ permaneceu o mesmo. A apoptose foi menor nas culturas de SCU. A expressão de integrina caiu, na população de células CD34+ e de CD45+ / Mononuclear cells from umbilical cord blood (UCB) and G-CSF mobilized peripheral blood (MPB), were cultured in vitro, in the presence of cytokines, with or without bone marrow stroma. The aims were to evaluate the proliferative response of progenitor cells, occurrence of apoptosis and expression of adhesion molecule. In cultures without stroma, cellularity increased 5-fold for UCB, but has not changed for MPB. The number of CD34+ cells has dropped in both culture. With stroma, total nucleated cells had a 7-fold increse (UCB) and a 2,3-fold (MBP), however, CD34+ cells number has not changed. Apoptosis was lower in UCB culture. The expression of integrin decreased, in the CD34+ and CD45+ population

Células mononucleares de sangue de cordão umbilical (SCU) e sangue periférico mobilizado (SPM) com G-CSF, foram cultivadas in vitro com citocinas, na presença ou não de estroma de medula óssea. Os objetivos foram avaliar a capacidade proliferativa de células progenitoras, a ocorrência de apoptose e expressão de integrina. Nas culturas sem estroma, a celularidade aumentou 5 vezes (SCU) e não se alterou nas de SPM. O total de células CD34+ caiu em ambas culturas. Com estroma, o total de células nucleadas aumentou 7 vezes (SCU) e 2,3 vezes (SPM). O total de células CD34+ permaneceu o mesmo. A apoptose foi menor nas culturas de SCU. A expressão de integrina caiu, na população de células CD34+ e de CD45+ / Mononuclear cells from umbilical cord blood (UCB) and G-CSF mobilized peripheral blood (MPB), were cultured in vitro, in the presence of cytokines, with or without bone marrow stroma. The aims were to evaluate the proliferative response of progenitor cells, occurrence of apoptosis and expression of adhesion molecule. In cultures without stroma, cellularity increased 5-fold for UCB, but has not changed for MPB. The number of CD34+ cells has dropped in both culture. With stroma, total nucleated cells had a 7-fold increse (UCB) and a 2,3-fold (MBP), however, CD34+ cells number has not changed. Apoptosis was lower in UCB culture. The expression of integrin decreased, in the CD34+ and CD45+ population