Made available in DSpace on 2017-07-10T14:17:04Z (GMT). No. of bitstreams: 1 Mayaha.pdf: 4416602 bytes, checksum: 8b663982eaff70076dc570733618b841 (MD5) Previous issue date: 2013-08-01 / Bombyx mori multiple nucleopolyedrovirus (BmMNPV) is an entomopathogenic virus of the Baculoviridaefamily, genera Alphabaculovirus, which infects the silkworm (Bombyx Mori) and causes nuclear polyhedrosis disease. This virus is poliorganotrophic and a series of tissues are known as targets; however, literature does not present information in regards to the pylorus, segment of the hindgut, present in the transition with the midgut and responsible for regulating the passage of food towards the ileum. The organ is, thus, of fundamental importance in the ending of the digestive process, affecting the insect s metabolic balance and, the present study sought to analyze its susceptibility and cytopathology in regards to BmMNPV, geographic isolate of Paraná, Brazil. Moreover, due to the complexity of this segment, its general morphology will also be analyzed. In order to do so, hybrid B.mori caterpillars at 5º instar were inoculated experimentally with a viral suspension of BmMNPV. On different day post-inoculation (dpi), from 2º to 9ºdpi, segments of the intestine, containing the pylorus, with its subdivisions (posterior interstitial ring, pyloric cone and pyloric valve), were dissected, following the routine histological processing for paraffin wax blockage and dyes in haematoxylin and eosin, for general morphology analysis, and in modified Azan staining, for cytopathology. Segments also processed to form scanning electronic microscope for analized of morphological details. So the pylorus general morphology the caterpillars of B. mori, was similar to that observed in other insects. The cytopathology showed that the anterior area of the posterior interstitial ring is a secondary target of infection for the BmMNPV, after 5º dpi, and its infectious cycle is similar to that described in literature. In the hypertrophic nucleus one there the formation of viroplasm, place for production of enveloped nucleocapsids or virions, following the formation and development of the polyhedra. Later on, cytolysis occurred, promoting the liberation of polyhedra and the spreading of the disease, mainly in the B. mori creation sheds. The posterior area of the posterior interstitial ring, the pyloric cone and the pyloric valve did not reveal any traces of infection by BmMNPV and, thus, resistance mechanisms involving genetic factors could be acting. The knowledge obtained from this work will contribute in the establishment of the infectious cycle of this important entomopathogenic virus, which may lead to an epizootic that can negatively affect the whole productive chain of silk / Bombyx mori multiple nucleopolyedrovirus (BmMNPV) é um vírus entomopatogênico da família Baculoviridae, gênero Alphabaculovirus, que infecta o bicho-da-seda (Bombyx mori) e causa a doença poliedrose nuclear. Este vírus é poliorganotrófico e uma série de tecidos são conhecidos como alvos; contudo, a literatura não apresenta informações sobre o piloro, segmento do intestino posterior, presente na transição com o médio e responsável pela regulação da passagem do alimento em direção ao ileo. O órgão é assim de fundamental importância na finalização do processo digestório, afetando o equilíbrio metabólico do inseto e, o presente estudo objetivou analisar sua susceptibilidade e citopatologia frente ao BmMNPV, isolado geográfico do Paraná, Brasil. Ainda, devido a complexidade deste segmento, também será analisada sua morfologia geral. Para tanto, lagartas híbridas de B. mori no 5° instar foram inoculadas experimentalmente com uma suspensão viral de BmMNPV. Em diferentes dias pós-inoculação (dpi), do 2° ao 9° dpi, segmentos do intestino, contendo o piloro, com suas subdivisões (anel intersticial posterior, cone pilórico e válvula pilórica), foram dissecados, seguindo o processamento histológico de rotina para emblocamento em parafina e colorações em hematoxina e eosina, na análise da morfologia geral, e em Azan modificado, para a citopatologia. Segmentos também foram processados em microscopia eletrônica de varredura para análise de detalhes morfológicos. Assim, a morfologia geral do piloro das lagartas da B. mori foi semelhante à observada em outros insetos. A citopatologia mostrou que a área anterior do anel intersticial posterior é alvo secundário de infecção pelo BmMNPV, a partir do 5° dpi, e seu ciclo infeccioso é semelhante ao descrito em literatura. No núcleo hipertrófico houve a formação do viroplasma, local de produção dos nucleocapsídeos envelopados ou virions, seguindo a formação e o desenvolvimento dos poliedros. Posteriormente, ocorreu a citólise, promovendo a liberação dos poliedros e disseminação da doença, principalmente nos barracões de criação de B. mori. A área posterior do anel intersticial posterior, o cone pilórico e a válvula pilórica não revelaram quaisquer indícios de infecção pelo BmMNPV e, desta forma, mecanismos de resistência envolvendo fatores genéticos poderiam estar atuando. Os conhecimentos obtidos irão contribuir no estabelecimento do ciclo infeccioso deste importante vírus entomopatogênico, cuja conseqüência de uma epizootia pode afetar negativamente toda a cadeia produtiva da seda

Introdução: A doença intersticial pulmonar é um importante fator prognóstico na esclerose sistêmica (ES). O prognóstico das pneumonias intersticiais não específicas (NSIP) associadas às colagenoses tem sido descrito como melhor do que o da forma idiopática. Levanta-se a hipótese de que o processo de remodelamento e reparo do parênquima pulmonar nessas duas formas da doença sejam diferentes. Objetivos: Comparar os mecanismos de reparo e remodelamento entre a NSIP associada a ES e a NSIP idiopática. Observar o impacto dos mesmos nas provas de função pulmonar e na sobrevida. Métodos: Foram analisadas 40 biópsias de pacientes com o diagnóstico de NSIP (18 biópsias de NSIP associada a ES e 22 na forma idiopática). As informações clínicas e as provas de função pulmonar foram obtidas através da revisão dos prontuários. As lâminas foram revisadas por três patologistas. Foram comparadas as densidades epitelial, vascular, bem como a atividade vascular dos dois grupos utilizando o método de imuno-histoquímica. Para isso foram utilizados os anticorpos anti- citoqueratina 7 (CK-7), anti- proteína-a do surfactante (SP-A), antimarcador de célula endotelial CD-34 (CD34), e anti-molécula da adesão vascular 1 (VCAM-1). Também foi comparado o padrão de remodelamento da matriz septal e vascular, usando os métodos histoquímicos da resorcina (fibras elásticas) e picrosírius (colágeno). Uma análise estatística foi realizada comparando os resultados dos dois grupos, o impacto do remodelamento nas provas de função pulmonar e a influência na sobrevida. Resultados: A densidade das células epiteliais foi menor na NSIP-ES, do que na forma idiopática (p<0,0001). Já os pneumócitos tipo II e as células de Clara encontraram-se diminuídos no grupo idiopático (p=0,02). Uma diminuição na densidade vascular foi encontrada na NSIP-ES quando comparada à forma idiopática (p<0,0001); no entanto, a atividade vascular medida pelo VCAM-1 foi maior no grupo da NSIP-ES (p<0.0001). O conteúdo das fibras elásticas e colágeno septal, bem como o das fibras elásticas na parede vascular, estavam aumentados no grupo da ES quando comparados à forma idiopática (p=0,01; p=0,001 e p<0,0001, respectivamente). Não houve diferença estatística entre o colágeno da parede vascular, no grau de obstrução vascular ou associação entre os parâmetros de remodelamento e reparo do parênquima pulmonar na sobrevida dos dois grupos. Dentre as provas de função pulmonar, a DCO/Hb foi mais afetada no grupo da ES (59% do valor predito na ES e 97% no grupo idiopático). Foi observada uma associação direta entre a densidade vascular e a DCO/Hb (p=0,02). Após o seguimento de 36 meses, não foi observada diferença no prognóstico dos dois grupos. Conclusão: Os processos de remodelamento e reparo do parênquima pulmonar parecem ser diferentes entre os dois grupos. Apesar de o processo fibrótico ser mais intenso na NSIP-ES, isso parece não estar associado a um pior prognóstico, como tem sido descrito na forma idiopática. Como o processo de elastose e a expressão do VCAM-I são mais intensos na ES, isso sugere que o processo inflamatório tem um papel mais importante na patogênese e no processo de remodelamento e reparo da ES do que na forma idiopática. No entanto, outros estudos são necessários para validar a importância desses resultados e sua utilização para fins terapêuticos e de prognóstico / Background: The presence of Interstitial lung disease is a well recognized prognostic factor in systemic sclerosis (SSc). As the prognosis in nonspecific interstitial pneumonia (NSIP) has been described to be better in collagen vascular disorders compared to the idiopathic forms, it is conceivable that the mechanisms of repair and remodeling are different between these two forms of the disease. Objectives: To compare the mechanisms of repair and remodeling between SSc associated nonspecific pneumonia and the idiopathic form, as well as their impact on pulmonary function tests and survival rates. Methods: Biopsies from 18 patients with SSc-associated NSIP and 22 with idiopathic NSIP were analyzed. Clinical data and pulmonary function test results were obtained by retrospective chart review. All H&E slides were reviewed by three pathologists. The epithelial and vascular densities and vascular activity were compared between the two groups by immunohistochemistry with antibodies directed against cytokeratin-7, surfactant protein-a, CD34, and VCAM-1, as well as septal and vascular matrix remodeling using histochemical stains (picrosirius and resorcin). Statistical analyses were performed to compare the results of these various studies with clinical parameters (e.g. pulmonary function tests) and survival between the groups. Results: Epithelial cell density was lower in SSc-NSIP when compared with idiopathic-NSIP (p<0.0001). Type II pneumocytes and Clara cells were reduced in idiopathic NSIP (p=0.02). A decrease in microvessel density was found in SSc-NSIP compared to idiopathic-NSIP (p<0.0001). The vascular activity measured by VCAM-1 expression was higher in NSIP-SSc when compared to the idiopathic group (p<0.0001). A direct association between vascular density and DLCO/HB was found (p=0.02). Among pulmonary function tests the DLCO/HB was affected to a greater extent in the SSc group (59% of the predicted value in SSc and 97% in the idiopatic group). The content of septal collagen and elastic fibers, as well as the elastic fibers in the vascular wall, were higher in the SSc group (p=0.01, p=0.001 and p<0.0001, respectively). There were no differences in the collagen content of the vascular wall, vascular grade, or survival between the two groups. There was no difference in the survival rate between the two groups after a follow-up of 36 months. Conclusions: Alterations in the pulmonary epithelium and vasculature seem to differ in the SSc-NSIP when compared to the idiopathic form of the disease. Although the fibrotic process is more intense in the SSc group, it does not seem to affect the prognosis of these patients, contrary to what has been described in idiopatic lung fibrosis. Because the elastotic process and VCAM-1 expression are higher in the SSc group, this might suggest that inflammatory mechanisms affecting the elastic fiber system and vasculature could play a greater role in the pathogenesis and pulmonary remodeling process of SSc-NSIP than in idiopathic-NSIP. Further studies may be required to assess the significance of these findings and explore if they can provide prognostic and/or treatment information

<p>Molecular mechanisms in the development and progression of thyroid carcinomas are still not fully understood. In the present thesis the highly malignant anaplastic thyroid carcinoma (ATC) was used to study regulation of apoptosis and tumor interstitial fluid pressure (IFP).</p><p>Addition of a natural estrogen metabolite, 2-Methoxyestradiol (2-ME), induced a G2/M cell cycle arrest and apoptosis in five out of six human ATC cell lines. Treatment with 2-ME induced DNA-fragmentation as well as activation of caspase-3. Inhibitors of JNK and p38 MAPKs activity decreased the effect of 2-ME suggesting involvement in the induction of apoptosis.</p><p>Solid tumors have an elevated IFP. High IFP forms or reflects a barrier for exchange of molecules between microvessels and surrounding tissue. The mechanisms for the generation of the high IFP were investigated using a specific TGF-β inhibitor in an ATC model in athymic mice. Tumor IFP was lowered in TGF-β inhibitor-treated compared to control mice. Affymetrix microarray analysis showed a decreased expression of macrophage-associated genes in treated tumors. Furthermore, the number and activity of tumor-associated macrophages was reduced after TGF-β inhibition. A decreased protein leakage together with an increased coverage of α-smooth-muscle actin (SMA)-expressing cells indicated vessel normalization. An adjuvant treatment with the TGF-β inhibitor resulted in an increased treatment efficacy of doxorubicin. Thus, TGF-β inhibitor-treatment suggests improved microvessel function which results in a lowering of tumor IFP and increased tumor drug uptake.</p><p>To create a model for specific inactivation of genes in the thyroid, a transgenic mouse with a thyrocyte-specific expression of Cre recombinase was generated. The thyroglobulin promoter together with an inducible Cre recombinase (<i>creER</i><i>T2</i>) was used. Two transgenic founder lines were identified expressing cre mRNA solely in the thyroid. Functional activity of the CreER^T2 protein was demonstrated by using a ROSA26-LacZ reporter mouse.</p>

The correlation between NMR chemical shifts of interstitial atoms and electronic structures of boron- and carbon-centered hexazirconium halide clusters was investigated by density functional theory (DFT) calculation. The influences of bridging halide and terminal ligand variations on electronic structure were examined respectively. Inverse proportionality was found between the chemical shifts and the calculated energy gaps between two Kohn-Sham orbitals of t1u symmetry, which arose from the bonding and antibonding interaction between the zirconium cage bonding orbitals and the interstitial 2p orbitals. Chemical shielding properties of the interstitial atoms were calculated with Gauge Including Atomic Orbital (GIAO) method. Stepwise ligand substitution of terminal chlorides on [(Zr6CCl12)Cl6]4-cluster by tri(n-butyl)-phosphine oxide (Bu3PO) was conducted with the aid of TlPF6. Composition of the reaction mixtures was analyzed by use of both 13C and 31P NMR. A preliminary scheme for synthesis and separation of [(Zr6CCl12)Cl6-x(Bu3PO)x]x-4 (x = 3 ?? 5) mixture based on solubility difference was reevaluated. Three 1,10-phenanthroline based bidentate ligands, namely, 2,9-Bis(diphenyl-phosphinyl)-1,10-phenanthroline, 2,9-Bis(diethoxyphosphoryl)-1,10-phenanthroline, and 2,9-Bis(di-n-butoxyphosphoryl)-1,10-phenantholine, were synthesized for bridge-chelating the hexazirconium clusters. Coordination chemistry of these ligands with the [Zr6BCl12] and [Zr6CCl12] clusters was subject to preliminary investigation.

Molecular mechanisms in the development and progression of thyroid carcinomas are still not fully understood. In the present thesis the highly malignant anaplastic thyroid carcinoma (ATC) was used to study regulation of apoptosis and tumor interstitial fluid pressure (IFP). Addition of a natural estrogen metabolite, 2-Methoxyestradiol (2-ME), induced a G2/M cell cycle arrest and apoptosis in five out of six human ATC cell lines. Treatment with 2-ME induced DNA-fragmentation as well as activation of caspase-3. Inhibitors of JNK and p38 MAPKs activity decreased the effect of 2-ME suggesting involvement in the induction of apoptosis. Solid tumors have an elevated IFP. High IFP forms or reflects a barrier for exchange of molecules between microvessels and surrounding tissue. The mechanisms for the generation of the high IFP were investigated using a specific TGF-β inhibitor in an ATC model in athymic mice. Tumor IFP was lowered in TGF-β inhibitor-treated compared to control mice. Affymetrix microarray analysis showed a decreased expression of macrophage-associated genes in treated tumors. Furthermore, the number and activity of tumor-associated macrophages was reduced after TGF-β inhibition. A decreased protein leakage together with an increased coverage of α-smooth-muscle actin (SMA)-expressing cells indicated vessel normalization. An adjuvant treatment with the TGF-β inhibitor resulted in an increased treatment efficacy of doxorubicin. Thus, TGF-β inhibitor-treatment suggests improved microvessel function which results in a lowering of tumor IFP and increased tumor drug uptake. To create a model for specific inactivation of genes in the thyroid, a transgenic mouse with a thyrocyte-specific expression of Cre recombinase was generated. The thyroglobulin promoter together with an inducible Cre recombinase (creERT2) was used. Two transgenic founder lines were identified expressing cre mRNA solely in the thyroid. Functional activity of the CreERT2 protein was demonstrated by using a ROSA26-LacZ reporter mouse.

The kidney regulates the excretion of water and electrolytes, which maintains homeostasis and enables control of arterial blood pressure. Hyaluronan, a large negatively charged interstitial glucosaminoglycan, is heterogeneously distributed within the kidney, primarily found in the medulla. Medullary hyaluronan content changes depending on the state of body hydration and plays a part in fluid regulation through its water binding and viscoelastic properties. The aim of this thesis was to provide new insight into the regulation of intrarenal hyaluronan during different states of body hydration, during completion of kidney development, and during diabetes mellitus. Dehydration reduces medullary interstitial hyaluronan in parallel with reduced hyaluronan synthase 2 gene expression and increased urinary hyaluronidase activity. Acute hydration results in an increase in medullary hyaluronan, an increase that requires nitric oxide and prostaglandins. Urinary hyaluronidase activity decreases during hydration. The elevation of hyaluronan is important for reducing water permeability of the interstitium i.e. favoring diuresis. Changes in hyaluronan concentration constitute a morphoregulatory pathway that plays a key role in nephrogenesis. The reduction in neonatal hyaluronan depended on an angiotensin II mediated process that does not appear dependent on lymph vessel formation. If angiotensin II is blocked with an ACE inhibitor, hyaluronan accumulates, which results in structural and functional abnormalities in the kidney. Renomedullary hyaluronan is elevated during uncontrolled diabetes, which coincides with induction of hyaluronan synthase 2 mRNA, hyperglycemia, glucosuria, proteinuria and overt diuresis. The levels of hyaluronan are probably at a terminus ad quem as no further response was seen during hydration. The higher interstitial expression of hyaluronan during diabetes may be involved in the progression of diabetic nephropathy. This thesis in physiology provides new mechanistic insights into the regulation of renal hyaluronan during various aspects of fluid handling.

In systemic sclerosis (SSc), interstitial lung disease (ILD) and engagement of the vascular system lead to increased morbidity and mortality. The aim of this thesis was to elucidate, in a consecutively included cohort of SSc (limited and diffuse) patients (n = 33), the T cell cytokine profile driving the disease in ILD and to explore the role of matrix metalloproteinase 9 (MMP-9) and its inhibitor: tissue inhibitor of metalloproteinase 1 (TIMP-1) in the extracellular matrix (ECM) degrading process leading to fibrous scarring and honey combing. Moreover, to characterize the role of nitric oxide (NO) in vascular engagement. Peripheral arterial changes cause Raynaud’s phenomenon and digital ulcers. Nitric oxide (NO) a main inducer of vasodilation is produced by endothelial nitric oxide synthase (eNOS) in response to changes in blood flow or by inflammatory cytokine inducible (i) NOS. In the vascular smooth muscle cell (VSMC) NO activates guanylate cyclase to produce cGMP, causing relaxation. We showed elevated plasma nitrate, a degradation product of NO, and increased urinary excretion of nitrate and cGMP. Plasma nitrate correlated with elevated levels of endothelial adhesion molecules: endothelial (E) selectin and vascular adhesion molecule 1, indicating that the activated endothelium is the site of NO synthesis by iNOS. Endothelial staining for E-selectin and the finding of iNOS and eNOS in SSc skin biopsies supported this notion. In SSc increased vascular stiffness may limit the NO vasodilatory effects. We found normal endothelium-dependent (i.e. flow mediated (FMD%)) and endothelium-independent (i.e. nitroglycerin-induced (NTG%)) vasodilation in the brachial artery. Radial arterial wall stiffness measured as maximum increase in pulse pressure (dP/dtmax) was increased. FMD% and especially NTG% correlated negatively and dP/dtmax positively to measures of endothelial inflammation: plasma- nitrate and adhesion molecule levels. Thus inflammatory vascular wall changes may interfere with dilation as may the presence of nitrate tolerance. We found elevated alveolar MMP-9 in both its pro- and active form in ILD. The levels correlated to decline in lung capacity, pointing at a causal relation. We suggest that neutrophils secrete MMP-9, which may degrade collagen IV, (the main constituent of basal membranes), collagen V, gelatins, proteoglycans and elastin. MMP-9 activity is partly regulated by the binding of pro- and active form to TIMP-1. Alveolar TIMP-1, which even stimulates fibroblast ECM synthesis, was increased independent of ILD. The inflammatory process in ILD is orchestrated by activated T helper (h) lymphocytes. We found a mixed Th1/Th2 reaction in SSc alveolar T cells expressing messenger for interferon gamma (Th1), IL-6 and IL-10 (both Th2). No particular cytokine mRNA profile distinguished alveolar T cells in ILD. Neutrophils invaded the bronchial epithelium, which seemed otherwise inert as levels of inflammatory cytokine sensitive transcription factors and their nuclear translocation tended to be low. The neutrophil recruitment pathway is uncertain as chemoattractants and endothelial adhesion molecules were normally expressed. In conclusion, MMP-9 probably causes degradation of lung tissue in ILD and may represent a future therapeutic target. Alveolar T cells show a mixed Th1/Th2 cytokine profile independent of ILD. Neutrophils invade the bronchial epithelium. Activated endothelium produces increased amounts of NO and adhesion molecules and the level of activation influences brachial arterial FMD% and NTG% and radial arterial compliance. Nitrate tolerance may be present.

The aortic valve maintains unidirectional blood flow between the left ventricle and the systemic circulation. When diseased, the valve is replaced either by a mechanical or a bioprosthetic heart valve, that carry issues such as thrombogenesis, long term structural failure, and calcification, necessitating the development of more structurally and biologically sufficient long-term replacements. Tissue engineering provides a possible avenue for development, combining cells, scaffolds, and biochemical factors to regenerate tissue. The overall goal of this dissertation was to create a foundation for the rational design of a tissue engineered aortic valve. The novel approach taken in this thesis research was to view each of the three leaflets as a laminate structure. The first three aims consider the leaflet as a laminate structure comprising of layers of collagen, elastin, and glycosaminoglycans (GAGs). In the first aim, the effect of GAGs on the tensile properties and stress relaxation in the leaflet was investigated, by removing GAGs through increasing amounts of hyaluronidase. A decrease in GAGs led to significantly higher elastic moduli, maximum stresses, and hysteresis in the leaflet. In the second aim, the 3D elastic fiber network of the leaflet was characterized using immunohistochemistry and scanning electron microscopy. This structure was found to have regionally varying thicknesses and patterns. In the third aim, a novel hydrogel-fiber composite design was proposed to match the anisotropy of the leaflet. This composite composed of aligned electrospun poly(ε-caprolactone) (PCL) within a poly(ethylene glycol) diacrylate (PEGDA) matrix. Surface modification and embedding of the PCL did not significantly alter the anisotropy or strength of the underlying PCL scaffold, providing the basis for an anisotropic, biocompatible scaffold. In the last aim, a novel co-culture model was designed using magnetic levitation as a layered structure of valvular endothelial cells and interstitial cells. This technique was used to create co-culture models within hours, while maintaining cell phenotype and function, and inducing extracellular matrix formation, as shown by immunohistochemical stains and their gene expression profiling. The overall result of this dissertation is a clearer understanding of the layered structure-function relationship of the aortic valve, and its application towards heart valve tissue engineering.

The correlation between NMR chemical shifts of interstitial atoms and electronic structures of boron- and carbon-centered hexazirconium halide clusters was investigated by density functional theory (DFT) calculation. The influences of bridging halide and terminal ligand variations on electronic structure were examined respectively. Inverse proportionality was found between the chemical shifts and the calculated energy gaps between two Kohn-Sham orbitals of t1u symmetry, which arose from the bonding and antibonding interaction between the zirconium cage bonding orbitals and the interstitial 2p orbitals. Chemical shielding properties of the interstitial atoms were calculated with Gauge Including Atomic Orbital (GIAO) method. Stepwise ligand substitution of terminal chlorides on [(Zr6CCl12)Cl6]4-cluster by tri(n-butyl)-phosphine oxide (Bu3PO) was conducted with the aid of TlPF6. Composition of the reaction mixtures was analyzed by use of both 13C and 31P NMR. A preliminary scheme for synthesis and separation of [(Zr6CCl12)Cl6-x(Bu3PO)x]x-4 (x = 3 ?? 5) mixture based on solubility difference was reevaluated. Three 1,10-phenanthroline based bidentate ligands, namely, 2,9-Bis(diphenyl-phosphinyl)-1,10-phenanthroline, 2,9-Bis(diethoxyphosphoryl)-1,10-phenanthroline, and 2,9-Bis(di-n-butoxyphosphoryl)-1,10-phenantholine, were synthesized for bridge-chelating the hexazirconium clusters. Coordination chemistry of these ligands with the [Zr6BCl12] and [Zr6CCl12] clusters was subject to preliminary investigation.

一向強調使用者自主性的網路媒體，卻也出現插播式廣告(Interstitial)這種強迫閱聽眾接收的形式，這種情形引發了研究者的興趣。相較於傳統媒體，已經賦予閱聽人部分自主權的情形底下，廣告主又想藉由形式的操弄強迫閱聽人接收，這可能引起的負面態度相對傳統媒體來說可能更為嚴重。因此，本研究首先欲探討插播式廣告的使用是否有其他優點，例如強迫接收的形式，會吸引更多注意的緣故而留下較深刻的印象，而讓廣告主在可能導致閱聽人反感的風險下仍願意使用。其二，插播式廣告的使用是否必然引起閱聽人產生被迫收看的感受，以及察覺到廣告主意欲操弄的意圖，進而降低說服效果。換言之，插播式廣告能否藉由提供閱聽人娛樂價值，因而避免負面態度的產生。 結果發現，相對橫幅廣告來說，插播式廣告的確能吸引較多的注意，但並不會留下較深刻的記憶。其次，插播式廣告是否會引起操弄意圖推論、強迫性感受，進而降低說服效果，關鍵在於如果廣告能夠提供娛樂價值，那麼閱聽人並不會做出操弄意圖推論，也不會感覺到自己是被迫收看廣告的，當然不會進一步降低廣告態度。反之，若不提供其他價值，則會導致閱聽人做出操弄意圖推論，以及產生強迫性感受，因而進一步降低廣告態度。 最後，我們以一個交換的觀點來貫穿整篇文章，決定閱聽人是否產生操弄意圖推論或強迫性感受，關鍵在於閱聽人付出以及得到了多少價值，如果能讓閱聽人感受到自己所獲得的價值多過所付出的代價，那麼閱聽眾是會原諒行銷者的操弄。 / Comparing with other forms of media, the internet empowers audience with more freedom to choose what to see. However, it also emerged a new type of advertisement called, interstitial, which is commonly known as pop up. This kind of internet advertising forces audience to see advertisement whether they want or not. This phenomenon interests me to see what kind of effect this it will create for viewers., Since the audience once had the privilege to choose what they wanted to see, one of which is now seemed to be taken away from them, will this create negative feelings of the audience? Firstly I would like to know if there’re any advantages for advertisers to use this kind of internet advertising under the risk of disfavor. I postulates that interstitial may draw more attention and leave more impression than a banner does because of its format. Secondly, interstitial will not necessarily cause intrusiveness and inferences of manipulative intent (IMI), if the advertisement can provide other form values such as information or entertainment. The result supports my hypothesis partially. Firstly, interstitial does draw more attention than a banner. However, except for making people more impressed, no other difference has been found. Secondly, if interstitial contains entertaining value, it will not make audience feel intruded, IMI. ,and hostile attitude toward that advertisement. Whereas it will cause intrusiveness and IMI if the interstitial advertisement forces audience to see it without having any rewards. In this case it will reduce the advertising attitude in the audience. Finally, we can use an ‘exchange of ideas’ to sum up the main idea of this paper. The audience will devote their efforts and time to processing the given message, if the interstitial can reward the audience with more than what they pay for. They will also forgive the advertisers for manipulating them. The key to decide whether the audience feels intruded or knows of IMI depends on how much the audience give and receive.