The vitamin D metabolite 1,25-dihydroxyvitamin D3 [1α,25(OH)2D3] plays an important role in the regulation of musculoskeletal growth and differentiation. 1α,25(OH)2D3 mediates its effects on cells, including chondrocytes and osteoblasts, through the classical nuclear 1α,25(OH)2D3 receptor. Additionally, recent evidence indicates that several cellular responses to 1α,25(OH)2D3 are mediated via a rapid, calcium-dependent membrane-mediated pathway. These actions of 1α,25(OH)2D3 can be blocked by antibodies to protein-disulfide isomerase family A, member 3 (Pdia3), indicating that it is part of the receptor complex; however, the pathway which is activated by this receptor is not fully understood. The overall goal of this thesis was to examine the roles of phospholipase A2 activating protein (PLAA) and calcium calmodulin-dependent kinase II (CaMKII) in 1α,25(OH)2D3 rapid membrane-mediated signaling. We further investigated the interaction between two pathways regulating growth plate cartilage and endochondral bone formation, 1α,25(OH)2D3 and Wnt5a, at the receptor complex level. Results indicated that PLAA was required for mediating 1α,25(OH)2D3 signal from Pdia3. Furthermore, CaM and CaMKII were identified as mediators of 1α,25(OH)2D3-stimulated PLAA-dependent activation of cPLA2 and PKCα, and downstream biological effects. Wnt5a and 1α,25(OH)2D3 are important regulators of endochondral bone formation. This study demonstrated that 1α,25(OH)2D3 and Wnt5a mediate their effects via similar receptor components in osteoblasts and chondrocytes suggesting that these pathways may interact.

Universiẗat, Diss., 2000--Düsseldorf.

Das Secosterid Vitamin D3 wird durch die Nahrung aufgenommen oder im Organismus synthetisiert, wobei eine Reaktion in der Haut durch einen photochemischen Prozess katalysiert wird.Durch zwei Hydroxylierungsschritte in Leber und Niere wird Vitamin D3 über 25(OH) Vitamin D3 zum aktiven 1,25(OH)2 Vitamin D3-Hormon. 1,25(OH)2 Vitamin D3 hat eine wichtige Funktion im Knochenstoffwechsel, es reguliert die Ca2+-Resorption im Dünndarm. Die 1,25(OH)2 Vitamin D3-Synthese in der Niere wird durch Parathormon (PTH) kontrolliert. Ist die Serum Ca2+-Konzentration niedrig, wird PTH ausgeschüttet und die 1a-Hydroxylase, das 25(OH) Vitamin D3-aktivierende Enzym, stimuliert. Das Prinzip der (Seco)steroid-Aktivierung und -Inaktivierung in glandulären Organen, wie Leber und Niere mit anschließender Freisetzung der aktiven Hormone und Transport zu den jeweiligen Zielgeweben gilt heute nicht mehr uneingeschränkt. Auch Einzelzellen sind in der Lage Steroid-modifizierende Enzyme, die Hydroxylasen und Dehydrogenasen, zu exprimieren. Monozytäre Zellen exprimieren das 1,25(OH)2 Vitamin D3-aktivierende und das -inaktivierende Enzym, die 1a-Hydroxylase und die 24-Hydroxylase. Sie sind somit in der Lage, 1,25(OH)2 Vitamin D3 zu sezernieren, welches parakrin auf Nachbarzellen wirken kann. In diesem Zusammenhang wurde die Expression und Regulation der 1a-Hydroxylase in peripheren Blutmonozyten (PBM) und monozytären THP1-Zellen untersucht. Durch Supplementation der Zellen mit dem Substrat 25(OH) Vitamin D3 konnte die Produktion an aktivem 1,25(OH)2 Vitamin D3-Hormon in PBM signifikant gesteigert werden. In PBM konnte im Gegensatz zum systemischen Ca2+-Stoffwechsel nur ein geringer Einfluss auf die 1a-Hydroxylase-Aktivität beobachtet werden. Durch RT-PCR-Amplifikation konnte eine Expression des PTH Rezeptors Typ 1 (PTHR1) in PBM und Dendritischen Zellen nachgewiesen werden. Ein weiterer Ligand des PTHR1 ist PTH related Protein (PTHrP), ein Faktor der die Tumorhyperkalzämie propagiert. Durch Markierungsexperimente mit fluoreszenz-markiertem PTHrP konnte gezeigt werden, dass PTHrP an die Zellmembran von PBM und Dendritischen Zellen bindet und in den Zellkern von Dendritischen Zellen transportiert wird. Im Rahmen dieser Arbeit wurde die Expression 1,25(OH)2 Vitamin D3-responsive Gene in Monozyten/Makrophagen untersucht. Die Expression der 24-Hydroxylase wird innerhalb der Differenzierung von myeloischen THP1-Zellen zu Makrophagen- bzw. Osteoklasten-ähnlichen Zellen transient induziert. Als weiteres 1,25(OH)2 Vitamin D3-responsives Gen wurde die Expression von Osteopontin (OPN) untersucht. OPN ist ein vor allem in Knochen vorkommendes Matrixprotein, das wesentlich an der Zelladhäsion beteiligt ist. OPN wird in THP1-Zellen im Zuge der Differenzierung zunehmend exprimiert. Durch immunhistochemische Untersuchungen konnte OPN in Granulomen von Morbus Crohn- und Leberschnitten detektiert werden. Es spielt hier eine wesentliche Rolle bei der Granulomentstehung. Die Thioredoxin Reduktase 1 (TR1) ist ein Selenoenzym, welches maßgeblich an der Reduktion von Disulfidbindungen in Proteinen beteiligt ist. Es moduliert Protein/Protein- und Protein/DNA-Interaktionen wie die Bindung der Transkriptionsfaktoren AP1 und NFkB an DNA-responsive Elemente. Die Expression der TR1 wird in THP1-Zellen im Rahmen der Differenzierung induziert und ist in differenzierten Zellen maximal. Aktivitätsmessungen deckten sich mit dieser Beobachtung. In peripheren Blutmonozyten steigt die TR-Aktivität alleine durch Adhäsion der Zellen an das Kulturgefäß und nach Behandlung mit 1,25(OH)2 Vitamin D3. Die Untersuchungen der vorliegenden Arbeit zeigten eine Abhängigkeit der TR-Aktivität vom Differenzierungsgrad der Zellen und der Supplementation des Mediums mit dem Spurenelement Selen. Die Expression weiterer Selenoproteine in monozytären Zellen wurde nachgewiesen. So konnten durch 75Selenit-Markierungsexperimente neun Selenoproteine in THP1-Zellen detektiert werden, von denen fünf sezerniert werden. Ein weiteres, in monozytären Zellen charakterisiertes Selenoprotein ist die zelluläre Glutathionperoxidase. Ihre Aktivität konnte in Selenit-supplementierten Zellen um das 70fache gesteigert werden. Die Kultivierung monozytärer Zellen unter Selenit-Supplementation beeinflusst die Funktion dieser Zellen wesentlich. So konnte beobachtet werden, dass die Anzahl an phagozytierenden, zu Makrophagen differenzierten THP1-Zellen nach Selenit-Supplementation abnahm, während die Phagozytoserate der einzelnen Zellen anstieg. Die erzielten Ergebnisse zeigen, dass monozytäre Zellen mit Komponenten des 1,25(OH)2 Vitamin D3 Stoffwechsels ausgestattet sind und aktives 1,25(OH)2 Vitamin D3-Hormon produzieren, sezernieren und inaktivieren können. Die lokale Kontrolle der 1,25(OH)2 Vitamin D3 Stoffwechsels ausgestattet sind und aktives 1,25(OH)2 Vitamin D3-responsiver Gene, wie die Expression des Selenoproteins TR1, das einen direkten Einfluss auf den Redoxstatus und den Abbau reaktiver Sauerstoffverbindungen in diesen und Nachbarzellen ausübt. / The secosteroid 1,25(OH)2 vitamin D3 is either taken up by our daily diet or it is formed by a photochemical prosess in the skin. In liver and kidney vitamin D3 is hydroxylated in two steps to 25(OH) vitamin D3 and the active hormone 1,25(OH)2 vitamin D3. 1,25(OH)2 vitamin D3 plays an important role in bone metabolism. It is a key regulatorof the resorption of Ca2+ in the intestine. In the kidney 1,25(OH)2 vitamin D3 synthesis is controlled by parathyroid hormone (PTH). When the concentration of serum Ca2+ is low, PTH is secreted and 1a-hydroxylase, the 25(OH) vitamin D3 activating enzyme is induced in kidney. The picture of (seco)steroid activation and inactivation in glandular organs, like the liver and kidney, and the release and transport of the activated hormone to the target tissues has been modified recently. Single cells are also able to express steroid-modifying enzymes like hydroxylases and dehydrogenases. Monocytes express the 1,25(OH)2 vitamin D3-activating and the inactivating enzyme, i.e. the 1a-hydroxylase and the 24-hydroxylase. Thus they are able to build and secrete 1,25(OH)2 vitamin D3 which can act on neighbouring cells in a paracrine way. In this context the expression and regulation of the 1a-hydroxylase in peripheral blood monocytes (PBM) and THP1 cells was investigated. By supplementation of cells with the substrate 25(OH) vitamin D3 the production of active 1,25(OH)2 vitamin D3 hormone could be enhanced significantly in PBM. In PBM only a slight influence of PTH on 1a-hydroxylase activity could be observed, in contrast to the regulation in systemic Ca2+-metabolism. An expression of PTH receptor type 1 (PTHR1) could be verified by RT-PCR from whole RNA isolated from PBM and dendritic cells. A further ligand of PTHR1 is PTH related protein (PTHrP), a factor which propagates the humoral hypercalcemia of malignancy. Labeling experiments with a fluorescently marked PTHrP showed clustered membrane staining of PBM and dendritic cells and a transport to the nucleus of dendritic cells. The expression of 1,25(OH)2 vitamin D3-responsive genes in monocytes/macrophages was investigated. 24-hydroxylase is induced transiently during the differentiation of myeloid THP1 cells to macrophages and osteoclast-like cells, respectively. Next, the expression of osteopontin (OPN), a further 1,25(OH)2 vitamin D3 responsive gene was studied. OPN is a matrix protein that is mainly found in bone, it carries a RGD-motive in its aminoacid sequence which can bind to integrins and is involved in cell adhesion. The expression of OPN is increased during differentiation of THP1 cells. By immunohistochemistry OPN could be detected in Crohn's disease and liver granulomas where it also plays an important role in granuloma formation. The thioredoxin reductase 1 (TR1) is a selenoenzyme that is mainly involved in the reduction of disulfide bonds of proteins. It modulates protein/protein and protein/DNA interactions like the binding of the transkription factors AP1 and NFkB to DNA-responsive elements. The expression of TR1 mRNA is induced during differentiation and is maximal in differentiated cells. Activity measurments parallel these observations. In PBM TR-activity is increased by the event of adhesion of cells to the culture dish and after treatment with 1,25(OH)2 vitamin D3. A dependence of TR-activity on the degree of differentiation of cells and the supplementation of the medium with the trace element selenium was observed. The expression of further selenoproteins in monocytic cells was investigated. In THP1 cells nine selenoproteins could be detected By labeling experiments with 75selenite. Five were found as secreted proteins in the culture supernatant. In monocytes cellular glutathione peroxidase (cGPx) is a well characterized selenoprotein. Activity could be increased 70fold by selenit supplementation. Under selenite supplementation the number of differentiated THP1 cells capable of phagocytosis was diminished while the rate of phagocytosis of single cells was enhanced. Taken together, the experiments clearly indicate that monocytic cells are equipped with the components of 1,25(OH)2 vitamin D3 metabolism and thus are capable of 1,25(OH)2 vitamin D3 hormone synthesis, secretion and turnover. Moreover, local control of 1,25(OH)2 vitamin D3 synthesis and inactivation directly regulates the expression of 1,25(OH)2 vitamin D3-responsive genes like the selenoprotein TR and thus even impacts on the cellular redox-status and defense against reactive oxygen species in these and neighbouring cells.

Express parcel carriers offer a wide range of guaranteed delivery times in order to separate customers who value quick delivery from those that are less time but more price sensitive. Such segmentation, however, adds a whole new layer of complexity to the task of optimizing the logistics operations. While many sophisticated models have been developed to assist network planners in minimizing costs, few approaches account for the interplay between service pricing, customer decisions and the associated restrictions in the distribution process. This paper attempts to fill this research gap by introducing a heuristic solution approach that simultaneously determines the ideal set of services, the associated pricing and the fulfillment plan in order to maximize profit. By integrating revenue management techniques into vehicle routing and eet planning, we derive a new type of formulation called service selection, pricing and fulfillment problem (SSPFP). It combines a multi-product pricing problem with a cycle-based service network design formulation. In order derive good-quality solutions for realistically-sized instances we use an asynchronous parallel genetic algorithm and follow the intuition that small changes to prices and customer assignments cause minor changes in the distribution process. We thus base every new solution on the most similar already evaluated fulfillment plan. This adapted initial solution is then iteratively improved by a newly-developed route-pattern exchange heuristic. The performance of the developed algorithm is demonstrated on a number of randomly created test instances and is compared to the solutions of a commercial MIP-solver. / Series: Schriftenreihe des Instituts für Transportwirtschaft und Logistik - Supply Chain Management

Faraday waves arise on the surface of a liquid in a container that is undergoing vertical periodic oscillations. We investigate two-dimensional Faraday waves in a long rectangular container, both theoretically and experimentally. Hysteresis occurs when both finite amplitude solutions and the flat surface solution are available. We derive a nonlinear model of a standing wave, extending the Lagrangian method of Miles (1976). The model is used to investigate hysteresis. It is found necessary to retain cubic damping, cubic forcing and the fifth-order conservative term in order to achieve agreement with experiments. The fifth-order conservative term was omitted from all previous studies of Faraday waves. Stable limit cycles are found to arise from this single-mode equation. We examine the structure of this new solution in detail, both analytically and numerically. We describe local bifurcations using a multiple time scales analysis and global bifurcations using Melnikov's method. The coefficients of linear and cubic damping are derived for a standing wave in a rectangular container by considering energy dissipation in the main body of the fluid (due to potential flow and streaming) and in boundary layers at the sidewalls and at the surface. Surface contamination, due to the presence of a thin viscoelastic surface film, creates a boundary layer at the surface which causes enhanced dissipation comparable to, or greater than, that caused by the boundary layers at the walls of the container. Three-mode interaction equations are used to model intermittency and complex modulations which are found to arise from a sideband instability mechanism similar to that of Eckhaus (1963) and Benjamin & Feir (1967). The role of cubic and fifth-order nonlinear terms on this instability mechanism is examined. Theoretical results are found to compare quite favourably with experimental data.

This paper presents the first cross-country empirical evidence on the determinants of participation in Fairtrade and the impact of the export of Fairtrade certified products on agricultural growth in low- and middle-income countries. Using the number of certified producer organizations per country in 2006-2010 as a proxy for Fairtrade exports, estimation results indicate a small but significantly positive effect on the growth rate of per capita value added in agriculture that is largest in upper middle income countries. Given the particularly poverty-reducing effect of agricultural growth, we find empirical evidence that Fairtrade certification is indeed able to deliver its core values, but misses to target the very poor. (author's abstract) / Series: Department of Economics Working Paper Series

Zugl.: Berlin, Freie Universiẗat, Diss., 2009.

Thesis (M.S.C.I.T.)--Regis University, Denver, Colo., 2006. / Title from PDF title page (viewed on May 25, 2006). Includes bibliographical references.

Submitted by CARINA GUIDI PINTO null (carina_guidi@hotmail.com) on 2016-02-26T12:56:51Z No. of bitstreams: 1 Dissertação Carina Guidi Pinto.pdf: 2764188 bytes, checksum: 30bbdd1a9fc0615c8dae9f321f8a83a1 (MD5) / Approved for entry into archive by Ana Paula Grisoto (grisotoana@reitoria.unesp.br) on 2016-02-26T18:59:28Z (GMT) No. of bitstreams: 1 pinto_cg_me_bot.pdf: 2764188 bytes, checksum: 30bbdd1a9fc0615c8dae9f321f8a83a1 (MD5) / Made available in DSpace on 2016-02-26T18:59:28Z (GMT). No. of bitstreams: 1 pinto_cg_me_bot.pdf: 2764188 bytes, checksum: 30bbdd1a9fc0615c8dae9f321f8a83a1 (MD5) Previous issue date: 2016-01-22 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Pró-Reitoria de Pesquisa (PROPe UNESP) / A ingestão de etanol compromete a estrutura do cérebro, apresentar efeitos bifásicos sobre a atividade motora, agindo como um estimulante ou depressor dependendo da dose ou a duração de utilização. Ele interfere na absorção e metabolismo de vitamina D3, o que se correlaciona com alguns distúrbios neurológicos e neuropsiquiátricos. Há relatos sobre a associação de etanol com alterações ósseas, incluindo baixos níveis de vitamina D3. Com base nisso, o objetivo deste estudo foi avaliar os efeitos em testes de comportamento da administração isolada de vitamina D3 ou a sua administração em associação com etanol, durante alcoolismo crônico. A fim de conseguir isso, foram utilizados dois grupos experimentais: ratos Wistar machos (n = 20) e ratos UChB linhagem machos (n = 20) (bebedores voluntários de etanol). Ambos os grupos foram divididos em dois subgrupos: Vitamina D3 - 12.5μg / kg / dia (500 UI) de colecalciferol (WV, n = 10, e UV, n = 10), e de controle (CC, n = 10, e UC, n = 10), durante um período de 75 dias. O peso corporal análises e testes de comportamento (reflexo de sobressalto acústico e campo aberto) foram realizados em 90 e 165 dias de idade. Além disso, os níveis de plasma de corticosterona foram medidos a 165 dias, sem diferença estatística entre os grupos experimentais. O grupo Wistar apresentou valores mais baixos ASR no momento final (Controle e completada), enquanto as percentagens PPI foram maiores no grupo inicial. No grupo UChB houve nenhuma diferença em percentagens PPI com o pré-estímulos utilizados. Quando as respostas ASR foram comparados entre os grupos (Wistar e UChB), o grupo UChB apresentaram menores amplitudes de ASR quando comparados com os animais do grupo de ratos Wistar. Em relação à PPI percentagens dos três estímulos pré utilizados no grupo UChB, as percentagens foram maiores em comparação com os animais do grupo de ratos Wistar, os valores ASR foi menor no grupo UChB. A presença de fezes e urina foi semelhante tanto nos momentos iniciais e finais, no teste de campo aberto, para todos os grupos experimentais. Considerando-se os outros parâmetros de campo aberto, diminuição da locomoção foi observado no grupo UChB no subgrupo vitamina D3, bem como no grupo de controlo final, em comparação com o controlo inicial. Quando os grupos Wistar e UChB foram comparados, este último apresentado taxas mais elevadas na locomoção. Os resultados obtidos estão em consonância com o efeito sedativo / depressivo de etanol sobre o sistema nervoso central, reduzindo a ASR de amplitude, com um aumento provável ao longo dos circuitos inibitórios que medeiam a PPI, a determinação do aumento na percentagem de inibição no grupo UChB, bem como um aumento na locomoção apresentada pelo grupo UChB no teste de campo aberto, caracterizada por uma diminuição no efeito aversivo ao novo ambiente. Além disso ressaltamos que a dose de vitamina D3 usado não teve efeito sobre o parâmetro de comportamento analisados, diferenças observadas neste grupo parecendo estar relacionada a se acostumar com a sonda gástrica. Este estudo descreve pela primeira vez na literatura as diferenças nas respostas ao teste acústico de sobressalto reflexo entre ratos Wistar e UChB, este último geneticamente predispostos a ingestão de etanol, uma linhagem adequada para ser usada em experiências compreendendo análises de comportamento em face da ingestão de etanol. / Ethanol intake compromises brain structure, presenting biphasic effects over motor activity, acting as a stimulant or depressor depending on the dose or duration of use. It interferes in vitamin D3 absorption and metabolism, what correlates to some neurologic and neuropsychiatric disorders. There are reports on the association of ethanol with bone alterations, including low levels of vitamin D3. Based on that, the objective of this study was to evaluate the effects in behavior tests of isolated administration of vitamin D3 or its administration in association with ethanol, during chronic alcoholism. In order to achieve that, two experimental groups were used: male Wistar rats (n=20), and UChB lineage male rats (n=20) (volunteer ethanol drinkers). Both groups were divided in two subgroups: Vitamin D3 – 12.5µg/kg/day (500 UI) of cholecalciferol (WV, n=10, and UV, n=10), and Control (WC, n=10, and UC, n=10), for a period of 75 days. Body weight analyses and behavior tests (acoustic startle reflex and open field) were conducted at 90 and 165 days of age. In addition to that, corticosterone plasma levels were measured at 165 days, with no statistical difference between the experimental groups. The Wistar group presented lower ASR values in the final moment (Control and supplemented), while the PPI percentages were higher in the initial group. In the UChB group there was no difference in PPI percentages with the pre-stimuli used. When the ASR responses were compared between groups (Wistar and UChB), the UChB group presented lower ASR amplitudes as compared to the animals of the Wistar group. In relation to PPI percentages in the three pre stimuli used in the UChB group, the percentages were higher as compared to the animals in the Wistar group, the ASR values been smaller in the UChB group. The presence of feces and urine was similar both in the initial and final moments, in the open field test, for all the experimental groups. Considering the other parameters of open field, decreased locomotion was noted in the UChB group in the vitamin D3 subgroup, as well as in the final control group as compared to the initial control. When the Wistar and UChB groups were compared, the latter presented higher rates in locomotion. The results obtained are consonant to the depressive/sedative effect of ethanol over the CNS, reducing ASR amplitude, with a likely enhancement over the inhibitory circuits which mediate PPI, determining the increase in inhibition percentage in the UChB group, as well as an increase in locomotion presented by the UChB group in the open field test, characterized by a decrease in the aversive effect to the new environment. In addition we emphasize that the dose of vitamin D3 used did not have effect over the behavior parameter analyzed, differences observed in this group seeming to be related to getting used to the gavage. This study describes for the first time in literature the differences in responses to the acoustic startle reflex test between Wistar and UChB rats, the latter genetically predisposed to ethanol intake, a lineage adequate to be used in experiments comprising analyses of behavior in face of ethanol intake. / PROPe/RENOVE: 0252/010/14

Submitted by Nayara Passos (nayara.passos@ufpe.br) on 2015-03-05T13:22:25Z No. of bitstreams: 2 Dissertação Vanessa Lira.pdf: 2499393 bytes, checksum: f5562e19dfa4f6d45f4c03183fb1b153 (MD5) license_rdf: 1232 bytes, checksum: 66e71c371cc565284e70f40736c94386 (MD5) / Made available in DSpace on 2015-03-05T13:22:25Z (GMT). No. of bitstreams: 2 Dissertação Vanessa Lira.pdf: 2499393 bytes, checksum: f5562e19dfa4f6d45f4c03183fb1b153 (MD5) license_rdf: 1232 bytes, checksum: 66e71c371cc565284e70f40736c94386 (MD5) Previous issue date: 2013 / CAPES / Pratylenchus coffeae, conhecida popularmente no Nordeste por nematoide das lesões do inhame-dacosta, apresenta diversidade morfológica e de virulência entre populações de diferentes localidades. Como consequência, têm sido descritas novas espécies, a exemplo de P. pseudocoffeae e P. jaehni, anteriormente consideradas P. coffeae. Outra questão de relevo é o difícil controle populacional do nematoide no campo. O objetivo da pesquisa foi inicialmente caracterizar taxonomicamente dez populações do nematoide das lesões do inhame da costa, coletadas em diferentes regiões produtoras do Estado de Pernambuco. Para tal, foram utilizadas análises morfométricas e moleculares. Os resultados mostraram que as dez populações eram similares entre si e que possuíam mais de 70% de similaridade com os dados da literatura referentes à P. coffeae. Quanto a análise molecular, todos os espécimes analisados apresentaram 100% de homologia para P. coffeae, considerando-se as sequências do GenBank. Com isto, confirmou-se a identidade do nematoide das lesões do inhame-da-costa no Nordeste como sendo P. coffeae. O segundo objetivo foi identificar reações de hospedabilidade, resistência, tolerância, intolerância e de susceptibilidade em três leguminosas e em cinco gramíneas, todas de valor comercial, em relação ao parasitismo de P. coffeae. Os resultados mostraram que as variedades leguminosas feijão macassar IPA- 206 e IPA- 207 e as gramíneas milho São José e o milho doce como maus-hospedeiros-resistentes e o sorgo granífero como não-hospedeiro. Todos os genótipos podem ser utilizados em planos de rotação de culturas para controle de P. coffeae.