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Saelee, Netnapa, Chadanat Noonin, Benjamas Nupan, Kingkamon Junkunlo, Amornrat Phongdara, Xionghui Lin, Kenneth Söderhäll e Irene Söderhäll. "beta-Thymosins and Hemocyte Homeostasis in a Crustacean". Uppsala universitet, Jämförande fysiologi, 2013. http://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-200355.
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Thymosin proteins are well known for their actin-binding activity. Thymosin beta 4 (T beta 4) has been associated with biological activities in tissue repair and cell migration via interaction with ATP-synthase in vertebrates, while the information of similar thymosin functions in invertebrates is limited. We have shown previously that ATP-synthase is present on the surface of crayfish hematopoietic tissue (HPT) cells, and that astakine 1 (Ast1, an invertebrate cytokine) was found to interact with this beta-subunit of ATP synthase. Here, we identified five different beta-thymosins from Pacifastacus leniusculus, designated Pl-beta-thymosin1-5. The two dominant isoforms in brain, HPT and hemocytes, Pl-beta-thymosin1 and 2, were chosen for functional studies. Both isoforms could bind to the b-subunit of ATP-synthase, and Pl-beta-thymosin1, but not Pl-beta-thymosin2, significantly increased extracellular ATP formation. Moreover, Pl-beta-thymosin1 stimulated HPT cell migration in vitro and Ast1 blocked this effect. Pl-beta-thymosin2 increased the circulating hemocyte number at an early stage after injection. Additionally, in vivo injection of Pl-beta-thymosin1 resulted in significant reduction of reactive oxygen species (ROS) production in crayfish HPT whereas Pl-beta-thymosin2 had a similar but transient effect. Both Pl-beta-thymosins induced the expression of Ast1 and superoxide dismutase (SOD) transcripts, while silencing of endogenous Pl-beta-thymosin 1 and 2 by RNAi resulted in significant reduction of the Ast1 and SOD transcripts. The diverse effects exhibited by Pl-beta-thymosin1 and Pl-beta-thymosin2 indicates that these proteins are involved in a complex interaction that regulates the hematopoietic stem cell proliferation and differentiation.
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Noonin, Chadanat. "Melanization and Hemocyte Homeostasis in the Freshwater Crayfish, Pacifastacus leniusculus". Doctoral thesis, Uppsala universitet, Jämförande fysiologi, 2013. http://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-209209.
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Blood cells or hemocytes play important roles in immunity. They are a major source of many immune-related molecules such as antibodies in adaptive immunity of vertebrates and prophenoloxidase (proPO) in invertebrates. In the crayfish Pacifastacus leniusculus, the proPO-system has been reported to be an important component of immune responses against microorganisms. In this study, several mutant strains of Aeromonas hydrophila were used to reveal that LPS (lipopolysaccharide) is an important factor for the pathogenicity of A. hydrophila, strongly inducing the proPO system and melanization. This proPO activating system is a multistep process, which has to be tightly controlled to avoid the harmful side effects of toxic intermediates. Many regulating factors have been reported to fine-tune the proPO-system. In this study, the cleavage of caspase-1-like activity was shown to be a novel negative regulator of PO activity in crayfish. Moreover, the fragments obtained by cleavage of proPO by the proPO-activating enzyme and caspase-1-like protein increased bacterial clearance. Thus, the peptides generated also have important biological functions. In addition to being a source of immune proteins, hemocytes also participate in phagocytosis, encapsulation, and nodulation. An infection normally causes a reduction of hemocyte numbers. Consequently, hemocyte homeostasis is important for maintaining appropriate hemocyte numbers in the circulation of the animal. This study shows that the reactive oxygen species level in the anterior proliferation center of crayfish hematopoietic tissue (HPT), together with cell proliferation, was increased during infection. Pl-β-thymosins were proposed to be involved in hemocyte homeostasis by increasing stem cell migration and thus increasing the circulating hemocyte number. Crayfish hemocyte numbers, as well astakine (Ast1 and Ast2) expression in hemocytes and HPT, were previously shown to be under circadian regulation. Here, we show that Ast1, Ast2, and proPO exhibit rhythmic expression in the crayfish brain similarly to their orthologs, prokineticin 1, prokineticin 2 and tyrosinase, respectively, in the zebrafish brain. Tyrosinase expression was detected in zebrafish brain cells while PO-positive cells were identified as hemocytes that had infiltrated into the crayfish brain. Therefore, this information suggests a close relationship between crayfish hemocytes and the crayfish brain as well as vertebrate neurons.
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Comber, Kate. "Investigation into the molecular mechanisms governing Drosophila embryonic hemocyte migration in vivo". Thesis, University of Bath, 2014. https://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.606669.
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Accumulating evidence highlights the importance of studying the migration of cells within the context of their natural environment as manipulating the substrate on which a cell is migrating can have a dramatic impact on the mode/mechanisms employed by cells during migration. Central to this phenomenon is the requirement of adhesion to the ECM in order to gain traction during migration. Integrins constitute the main family of cell receptors involved in mediating cell-ECM interactions during motility. Whilst traditionally two-dimensional cell culture studies have placed emphasis on the importance of these receptors for spreading and migration, it has become evident that within more confined environments these receptors, at least for some cell types, are less crucial. In this research we utilise Drosophila embryonic hemocytes as an in vivo model for cell migration. We show that whilst hemocytes migrate within confined environments in vivo, these cells depend on integrins for powering both developmental and inflammatory migrations. Given the close association between these receptors and the actin cytoskeleton we were surprised to discover that removal of the main β integrin subunit, Myospheroid, did not affect cell spreading in vivo and had only a small impact on lamellipodial structure and dynamics. Furthermore we discovered that, in contrast to other cell types previously analysed, removal of this integrin subunit in hemocytes was not accompanied by an increase in the rate of actin retrograde flow within the protrusions, which we believe could reflect abrogation of a positive feedback between Rho, ROCK and Myosin II contraction. Instead, we discover a key role for integrins in regulating the microtubule cytoskeleton, in the maintenance of a polarised microtubule bundle, termed a ‘microtubule-arm’. Although the molecular mechanisms by which this stabilisation is coordinated have yet to be identified, this provides important insight into the co-regulation of adhesion and microtubule cytoskeleton important for the migratory behaviour of these cells. Cell migration reflects the complex and integrated regulation of the actin cytoskeleton by diverse families of actin regulatory proteins. Using hemocytes as a model system, we also explore the regulatory interactions between two main actin regulatory proteins, Diaphanous and Enabled, in vivo. Whilst the function of these proteins in the formation of filopodial protrusions is overlapping, recent research has highlighted the ability of these proteins to regulate the activity of one another. We find that co-expression of Enabled in hemocytes is able to rescue the morphological and migratory defects resulting from overexpression of active Diaphanous. Thus, data here presents Enabled as a negative regulator of Diaphanous, which may play an important role in the migration of hemocytes in vivo.
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Luce-Fedrow, Alison. "Drosophila melanogaster as a model for studying Ehrlichia chaffeensis infections". Diss., Kansas State University, 2010. http://hdl.handle.net/2097/11969.
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Doctor of PhilosophyDepartment of Biology
Stephen Keith Chapes
Ehrlichia chaffeensis is an obligate, intracellular bacterium that causes human monocytic ehrlichiosis (HME). The bacteria are vectored by the Lone Star tick (Amblyomma americanum), which is found primarily in the Midwestern and Southeastern United States E. chaffeensis was first reported in 1986 and HME was designated a nationally reportable disease by the United States Centers for Disease Control in 1999. Ehrlichia grows in several mammalian cell lines, but NO consensus model for pathogenesis exists for arthropods or vertebrates. Moreover, the host genes required for intracellular growth of this bacteria are unknown. We first established that the bacteria could infect and replicate both in vitro and in vivo in Drosophila melanogaster S2 cells and adult flies, respectively. We performed microarrays on S2 cells, comparing host gene expression between permissive or non-permissive conditions for E. chaffeensis growth. A total of 210 permissive, exclusive and 83 non-permissive, exclusive genes were up-regulated greater than 1.5-fold above uninfected cells. We screened flies mutant for genes identified in our microarrays for their ability to support Ehrlichia replication. Five mutant stocks were resistant to infection with Ehrlichia (genes CG6479, separation anxiety, CG3044, CG6364, and CG6543). qRT-PCR confirmed that bacterial load was decreased in mutant flies compared to wild-type controls. In particular, gene CG6364 is predicted to have uridine kinase activity. Thus, the in vivo mutation of this gene putatively disrupts the nucleotide salvage pathway, causing a decrease in bacterial replication. To further test the function of gene CG6364 in bacterial replication, we obtained cyclopentenyl cytosine (CPEC) from the National Cancer Institute. CPEC is a cytidine triphosphate (CTP) inhibitor known to deplete CTP pools in various cancers and to exhibit antiviral activity. Consequently, it inhibits de novo nucleotide synthesis, but doesn’t affect the nucleotide salvage pathway. When S2 cells were treated with CPEC and infected with Ehrlichia, an increase in bacterial replication was confirmed by qRT-PCR. Furthermore, addition of cytosine to S2 cells also resulted in increased bacterial replication. Therefore the nucleotide salvage pathway through cytidine appears necessary for bacterial replication. Our approach has successfully identified host genes that contribute to the pathogenicity of E. chaffeensis in Drosophila.
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Arteaga, Blanco Luis Andres. "Differential cellular immune response of hemocyte of Galleria mellonella larvae against Actinobacillus pleuropneumoniae strains". Universidade Federal de Viçosa, 2016. http://www.locus.ufv.br/handle/123456789/9267.
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Os insetos respondem à infecção através da montagem de reações imunes do tipo celular e humoral. Os reguladores primários dessas respostas são células chamadas hemócitos, os quais medeiam importantes respostas celulares, incluindo a fagocitose, encapsulamento, nodulação, e também segregam fatores humorais, tais como opsoninas, fatores de melanização e peptídeos antimicrobianos. Os hemócitos circulam ao longo da hemocele (cavidade corporal do inseto) pelo fluxo rápido da hemolinfa (sangue), além disso, partes desses hemócitos também existem como células sésseis que estão associados aos tecidos. As larvas de Galleria mellonella são uma alternativa viável para os modelos tradicionais dos mamíferos para estudar a eficácia de drogas antimicrobianas e a patogênese de microrganismos in vivo. No entanto, apesar da sua importância como um modelo de infecção, aspectos biológicos sobre as células do sistema imunológico, tais como a densidade e dinâmica dos hemócitos das larvas são mal compreendidos. No presente trabalho, investigamos a resposta imune celular dos hemócitos circulantes das larvas de G. mellonella contra diferentes cepas da bactéria Gram-negativa Actinobacillus pleuropneumoniae: baixa virulência (780), alta virulência (1022), e cepas de referência do sorotipo 8 (R8). Os hemócitos foram classificados com base no seu tamanho, morfologia, coloração e seus papeis na resposta imune, incluindo cinco tipos: prohemócitos, plasmatócitos, granulócitos, esferulócitos, e oenocitóides. Contagem total de hemócitos, contagem diferencial de hemócitos, atividade dos fagolisossomos, resposta autofágica, viabilidade celular, e a ativação da caspase-3 (como indicador de apoptose) foram determinados em hemócitos circulantes provenientes de larvas desafiadas e controle. Demostramos pela primeira vez no modelo de G. mellonella que os plasmatócitos e granulócitos ativam suas respostas autofágicas através da formação dos autofagossomos após o contato com A. pleuropneumoniae. Além disso, nossos dados demonstram que a imunidade celular do presente modelo de infecção muda dependendo do grau de virulência das cepas bacterianas.
Insects respond to infection by mounting cellular and humoral immune reactions. The primary regulators of these immune responses are cells called hemocytes, which mediate important cellular immune responses including phagocytosis, encapsulation, nodulation and also secrete immune factors such as opsonins, melanization factors and antimicrobial peptides. Hemocytes circulate through the hemocoel (body cavity) by the swift flow of hemolymph (blood), and part of these hemocytes population are sessile and are attached to tissues. Larvae of Galleria mellonella is a widely used factitious host as a viable alternative to traditional mammalian models to study the efficacy of antimicrobial drugs and the microbial pathogenesis in vivo. However, despite their importance as an infection model, biological aspects about the immune cells, such as density and hemocyte dynamic of larvae are poorly understood. In the present study, we investigated the cellular immune response of hemocytes from G. mellonella larvae against three strains of the gram-negative bacterium Actinobacillus pleuropneumoniae: low virulent (780), high virulent (1022), and the serotype 8 reference strain (R8). Five types of larval hemocytes, prohemocytes, plasmatocytes, granulocytes, oenocytoids, and spherulocytes, were distinguished according to size, morphology, detection by molecular probes, dye-staining properties, and their role in the immune response. Total hemocyte count, differential hemocyte count, lysosome activity, autophagic response, cell viability, and caspase-3 activation were determined in circulating hemocytes of naïve and infected larvae. Granulocytes and plasmatocytes were the major hemocyte types involved in the cellular defense against A. pleuropneumoniae; these hemocytes activated phagolysosome activities associated with an autophagic response against the bacteria. Moreover, our results showed that apoptosis in circulating hemocytes after exposure to virulent bacterial strains was related to an excessive autophagic cell death response induced by stress and subsequent caspase-3 activation.
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Krzemien, Joanna. "Control of larval hematopoiesis in Drosophila ; microenvironment, precursors and cell lineage". Toulouse 3, 2008. http://www.theses.fr/2008TOU30206.
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L'hématopoiése larvaire de la drosophile a lieu au sein d'un organe spécialisé, la glande de la lymphe (LG) qui produit des plasmatocytes spécialisés dans la phagocytose et des cellules à cristaux nécessaires à la mélanisation des corps étrangers (4). La LG est aussi à l'origine des lamellocytes nécessaires à l'encapsulation de gros corps étrangers et qui se différencient en réponse à des challenges immuns particuliers tels que le parasitisme par des hyménoptères. En 2004, notre laboratoire a montré que Collier (Col), l'orthologue du facteur de transcription mammifère Early B-Cell Factor est exprimé et requis dans un petit groupe de cellules spécialisées de la LG, le PSC, pour la différenciation des lamellocytes en réponse au parasitisme. Outre le PSC, la LG est organisée en deux zones distinctes : une zone médullaire (MZ) contenant les cellules précurseurs et une zone corticale (CZ) formée des cellules différenciées ayant quitté la MZ. Au cours de la première partie de ma thèse, j'ai étudié le contrôle de l'hématopoièse larvaire et montré que le PSC régulait de manière cellulaire non autonome, l'activité de la signalisation JAK/STAT dans les cellules précurseurs, empêchant ainsi leur différenciation prématurée et préservant leur capacité à se différencier en lamellocytes. Le rôle clef du PSC dans le maintien d'un pool de progéniteurs l'apparente à la niche hématopoiétique des vertébrés, un micro-environnement cellulaire requis pour le maintien de cellules souches tout au long de la vie adulte. Ceci posait la question de l'existence de cellules souches hématopoiètiques chez la drosophile, une question que j'ai abordée dans la deuxième partie de ma thèse. L'utilisation de marqueurs de cellules souches, de marqueurs de division asymétrique, ou la recherche de cellules quiescentes ne m'ont pas permis d'identifier des cellule souches dans la LG. J'ai alors mis en oeuvre des expériences de lignage cellulaire et montré que les pro-hémocytes acquièrent un destin plasmatocyte ou cellule à cristaux dés le premier stade larvaire, avant une phase de prolifération intense. .The Drosophila larval hematopoietic organ, the lymph gland (LG), disperses at metamorphosis, releasing two types of hemocytes: plasmatocytes involved in phagocytosis and crystal cells necessary for encapsulation which differentiate in response to specific immune challenges, such as parasitization by wasps. Collier (Col), the Drosophila ortholog of mammalian Early B - Cell Factor, is expressed and required in a small group of cells of the LG, the PSC and in the differentiation of lamellocytes. In addition to the PSC, the LG is organised in two zones: a medullary zone (MZ) containing prohemocytes and a cortical zone (CZ) containing differentiating cells. I showed that the PSC controls the balance between the pool of prohemocytes and differentiating hemocytes. PSC cells act, in a non-cell autonomous manner, to maintain JAK/STAT signaling in prohemocytes, preventing their premature differentiation and preserving the multipotent character necessary for lamellocyte differentiation. The PSC acts a micro-environment for Drosophila hematopoietic precursors which is reminiscent of the HSC (Hematopoietic Stem Cell) niche of vertebrates. To see if there exist HSC in the LG, I looked for stem cell markers and I could't get evidence for the presence of bona fide stem cells in the LG. Using clonal analyses, I determined that hemocyte precursors become committed to either plasmatocytes or crystal cells durind the L1 larval stage, followed by a phase of intense proliferation. Finally, I obtained evidence that lamellocytes and crystal cells share a common progenitor
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Hall, Jonathon Michael. "Temporal changes in the fatty acid composition and fluidity of gill and hemocyte membranes during thermal acclimation of the sea scallop, Placopecten magellanicus". Thesis, National Library of Canada = Bibliothèque nationale du Canada, 1999. http://www.collectionscanada.ca/obj/s4/f2/dsk2/ftp01/MQ54894.pdf.
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Liu, Haipeng. "Functional Studies of Some Immune Relevant Genes in a Crustacean". Doctoral thesis, Uppsala universitet, Jämförande fysiologi, 2008. http://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-9194.
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The freshwater crayfish, Pacifastacus leniusculus, mounts a strong innate immune response against microbes such as viruses and bacteria. In this thesis, a novel RNA interference (RNAi) method mediated with histone H2A was developed and applied in crayfish hematopoietic tissue cell cultures for gene functional studies. Further, the interactions between host (crayfish) and pathogens (white spot syndrome virus and Aeromonas hydrophila, respectively) were studied using RNAi technology in live animals. An antilipopolysaccharide factor isolated from viral challenged crayfish by suppression subtractive hybridization was shown to interfere with the propagation of white spot syndrome virus both in vivo and in vitro in crayfish, suggesting an important role of this factor in antiviral defense. Besides, RNAi of phenoloxidase, a critical immune effector involved in melanization, revealed that phenoloxidase activity is necessary for crayfish immune defense against a highly pathogenic bacterial infection in crayfish. In addition, RNAi was also employed to study a marker protein gene involved in hemocyte maturation in crayfish. Taken together, these studies may provide more insights into the immune responses against pathogen invasion as well as hemocyte ontogenesis in crustaceans.
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Hart, Courtney. "THE EFFECTS OF 4-NONYLPHENOL ON THE IMMUNE RESPONSE OF THE PACIFIC OYSTER, CRASSOSTREA GIGAS, FOLLOWING BACTERIAL INFECTION (VIBRIO CAMPBELLII)". DigitalCommons@CalPoly, 2016. https://digitalcommons.calpoly.edu/theses/1609.
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Endocrine disrupting chemicals (EDCs) are compounds that can interfere with hormone signaling pathways and are now recognized as pervasive in estuarine and marine waters. One prevalent EDC in California’s coastal waters is the xenoestrogen 4-nonylphenol (4-NP), which has been shown to impair reproduction, development, growth, and in some cases immune function of marine invertebrates. To further investigate effects of 4-NP on marine invertebrate immune function we measured total hemocyte counts (THC), relative transcript abundance of immune-relevant genes, and lysozyme activity in Pacific oysters (Crassostrea gigas) following bacterial infection. To quantify these effects we exposed oysters to dissolved phase 4-NP at high (100 μg l-1), low (2 μg l-1), or control (100 μl ethanol) concentrations for 7 days, and then experimentally infected (via injection into the adductor muscle) the oysters with the marine bacterium Vibrio campbellii. 4-NP significantly altered the effects of bacterial infection had on THC. Oysters exposed to both high and low 4-NP did not experience a bacteria-induced increase in THC, as seen in control oysters. We also determined that V. campbellii infection induced differential expression of a subset of immune-related genes tested (Cg-bigdef2, Cg-bpi1, Cg-lys1, Cg-timp) in some, but not all, tissues; 4-NP exposure altered expression patterns in two of these genes (Cg-bpi1 and Cg-tgase). Exposure to 4-NP alone also caused differential expression in some genes (Cg-bpi1, Cg-galectin1, Cg-clec2). Lastly, low levels of 4-NP significantly increased lysozyme activity 24 h post-infection. These results suggest that exposure to 4-NP can alter both cellular and humoral immune responses to bacterial infection in C. gigas.
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BORELLO, ALESSIO. "Vibrio interactions with bivalve hemocytes and analysis of the Crassostrea gigas microbiota". Doctoral thesis, Università degli studi di Genova, 2021. http://hdl.handle.net/11567/1047238.
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My PhD project aimed at investigating the molecular mechanisms at the basis of the interaction between Vibrio bacteria and shellfish in the bivalve models Crassotrea gigas and Mytilus galloprovincialis and to study the composition and dynamics of bivalve microbiota.
Previous studies suggested that persistence of entrapped bacteria inside bivalve tissues depends, at least in part, on their capacity to survive to the hemolymph bactericidal activity, that is exerted by both hemocytes and serum soluble factors. In the first part of my PhD work, hemocytes of M. galloprovincialis were challenged with different pathogenic Vibrio strains (V. aestuarianus 01/032, V. aestuarianus 02/041, V. tasmaniensis LGP32, V. harveyi VH2, V. tapetis CECT 4600 and V. coralliilyticus ATCC BAA 450) in the presence or in the absence of the extrapallial protein present in M. galloprovincialis serum (MgEP), and of the whole hemolymph serum. In addition, C. gigas hemocytes were exposed to the bivalve pathogens V. aestuarianus 01/032 and V. aestuarianus 02/041 under the same conditions to better understand molecular basis of bacteria-hemolymph interactions in oysters. We observed that MgEP promotes D- mannose sensitive adhesion to and killing by hemocytes of the bivalve pathogens V. aestuarianus 01/032, V. aestuarianus 02/041, V. tasmaniensis LGP32 and V. coralliilyticus ATCC BAA 450. In addition, in the presence of M. galloprovincialis EP protein (MgEP), C. gigas haemocytes killed V. aestuarianus 01/032 and V. aestuarianus 02/041 almost as efficiently as mussel phagocytes. These findings suggest that the different sensitivity of Vibrio strains to the antibacterial activity of oyster (susceptible to Vibrio infection) and mussel (resistant to Vibrio infection) haemolymph might partly depend on the fact that C. gigas serum lacks MgEP-like opsonins. These results may have important implications for improving bivalve depuration strategies and prevent diseases affecting bivalve production worldwide.
In the second part of my thesis work, I studied the microbial communities associated to contrasting C. gigas samples collected during mortality episodes in different European sites. Real-time PCR targeting oyster pathogens (e.g. Ostreid herpesvirus 1 [OshV-1] and V. aestuarianus) and 16SrRNA gene-based microbial profiling were applied on a large number of C. gigas samples (n=525 and n=101 for qPCR and 16SrRNA gene profiling analysis, respectively) to extensively investigate the patterns and dynamics of oyster microbiota during mortality events. Comparative analysis of contrasting (e.g. infected vs not infected) C. gigas samples conducted using these methods revealed that oyster experiencing mortality outbreaks displayed signs of microbiota disruption associated with the presence of previously undetected potential pathogenic microbial species mostly belonging to genus Vibrio and Arcobacter. This represents to our knowledge, the largest study conducted so far to determine the composition and dynamics of farmed oyster microbiota.
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Piñera, Angelica Vivas. "The maturation of the immune system and the effects of crowding and light stress during development on the immune function of the adult house cricket Acheta domesticus". Miami University / OhioLINK, 2012. http://rave.ohiolink.edu/etdc/view?acc_num=miami1344570343.
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Bergum, Signe. "Combined Effects of Titanium Dioxide Nanoparticles (TiO2NPs) and Benzo(a)pyrene (B(a)P) on Hemocyte Cells and NADPH Cytochrome C Reductase Activity in Blue Mussels (Mytilus edulis)". Thesis, Norges teknisk-naturvitenskapelige universitet, Institutt for biologi, 2013. http://urn.kb.se/resolve?urn=urn:nbn:no:ntnu:diva-23625.
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Nanotechnology is currently one of the fastest growing industries. Titanium dioxide nanoparticles (TiO2NPs) are estimated to be one of the main produced engineered nanoparticles (ENPs) in the world today, and production is expected to increase. This might lead to enhanced emissions of ENPs to the environment. ENPs can be toxic to organisms. In addition, ENPs are seen to interact with pollutants and are thus assumed to alter the uptake of pollutants into organisms. Soil and sediments are suggested to be the main sink for ENPs together with other pollutants like polycyclic aromatic hydrocarbons (PAHs). As a consequence, benthic organisms are vulnerable to toxic effects of ENPs and for increased uptake of other pollutants by facilitated transportation of ENPs into the organisms. In the present study, blue mussels (Mytilus edulis) were exposed to titanium dioxide nanoparticles (TiO2NPs) (0.2 mg/L and 2.0 mg/L) and benzo(a)pyrene (B(a)P) (20 µg/L) singly and in combination with each other, for 96 hours. Cell damage on hemocytes and effects on the NADPH cytochrome c reductase activity in the digestive gland were examined with micronucleus assay, and by investigating the NADPH cytochrome c reductase activity with a spectrophotometric method. Protein concentrations in the mussels were determined by the Bradford method for normalizing the NADPH cytochrome c reductase activity.In this study, both B(a)P and TiO2NP increased the micronucleus (MN) frequency in hemocyte cells, both alone and in combination. A slight increase in MN frequency were detected from B(a)P exposed mussels to TiO2NP exposed mussels, and B(a)P+TiO2NP showed to have highest genotoxic effect in hemocyte cells. This suggests that both compounds have damaged the cell nucleus. No effects were detected for NADPH cytochrome c reductase activity normalized with the protein concentrations. However, protein concentrations increased from B(a)P exposed mussels to B(a)P+TiO2NP exposed mussels and the same was found for NADPH cytochrome c reductase activity without normalization for protein concentration. The findings of this study showed that TiO2NPs have genotoxic effects on mussel hemocytes, and that the toxic effect of a combined exposure of B(a)P and TiO2NP was higher than the effects of the single components. TiO2NP did not have toxic effects on the NADPH cytochrome c reductase activity. However, in combination with B(a)P, TiO2NP might adsorb B(a)P and transport the contaminant into mussels, and released B(a)P, which can have increased the NADPH cytochrome c reductase activity in the digestive gland of mussels. Further research is needed for a better understanding of TiO2NP toxicity on aquatic organisms, especially to elucidate interactions between ENPs and pollutants.
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Levin, David Michael. "An integrin required for the encapsulation immune response in the tobacco hornworm, Manduca sexta L. (Lepidoptera: Sphingidae)". Diss., Manhattan, Kan. : Kansas State University, 2007. http://hdl.handle.net/2097/412.
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Moreau, Pierrick. "Étude des interactions entre infection à ostreid herpesvirus 1, immunité, autophagie et pesticides chez l’huître creuse, Crassostrea gigas". Thesis, La Rochelle, 2014. http://www.theses.fr/2014LAROS028/document.
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Le travail de thèse s’inscrit dans la problématique très actuelle des mortalités massives de naissain et de juvéniles d’huîtres creuses, C. gigas, et des questionnements autour de l’implication des pesticides dans ce phénomène. La première partie de la thèse a été consacrée à l’étude des effets de pesticides sur les capacités hémocytaires de l’huître creuse (in vitro et in vivo). L’effet immuno-modulateur des xénobiotiques sélectionnés (seul ou en mélange) a été exploré principalement au travers de la cytométrie en flux. La deuxième partie de la thèse a concerné l’étude des effets d’un mélange de pesticides sélectionnés sur le virus OsHV-1 et l’infection qu’il induit chez l’huître creuse. Les effets des pesticides sur le virus lui-même ont été évalués. Les expériences réalisées n'ont pas permis de mettre en évidence, dans les conditions testées, de dégradation des particules virales en présence des polluants. D’autre part, leurs effets sur les huîtres elles-mêmes ont été explorés après traitement des animaux avec un mélange de pesticides lors d’essais de pathologie expérimentale. Il a ainsi pu être montré que des huîtres préalablement contaminées par un mélange de pesticides à des concentrations retrouvées dans l’environnement semblaient être plus sensibles à l’infection virale. La troisième partie de la thèse a concerné l’étude de l’autophagie chez l’huître creuse, C. gigas. La publication du génome complet de cette espèce en 2012 a ouvert de nouvelles possibilités pour étudier l'immunité innée. L’étude de l’autophagie réalisée pour la première fois chez l’huître creuse a consisté lors d’une première étape en la recherche in silico de gènes impliqués dans cette voie et des protéines correspondantes par western blotting. Puis, le rôle de ce processus important dans l’immunité innée a été exploré au travers d’essais de reproduction d’infections en présence ou non de modulateurs de l’autophagie. Les résultats obtenus semblent montrer que l’autophagie soit un processus important pouvant être impliqué dans les capacités de défense de l’huître creuse vis-à-vis d’infections virales et bactériennesThe thesis work is part of the very current issue on mass mortality outbreaks affecting Pacific oyster, C. gigas, spat and juveniles and questions about the involvement of pesticides in this phenomenon. The first part of this thesis was devoted to study the effects of pesticides on hemocyte parameters in the Pacific oyster (in vitro and in vivo). The immunomodulator effect of selected pesticides (alone or in mixture) has been explored principally through flow cytometry. The second part concerned the study of the effects of a pesticides mixture on OsHV-1 itself. No direct effects have been reported on the viral particles in presence of the pollutants. Moreover, pesticide effects on Pacific oysters were also explored through experimental pathology assays after treatment of animals with a polluant mixture. Results showed that pesticide treated oysters appeared more susceptible to the viral infection in experimental conditions. The third part concerned the study of autophagy in the Pacific oyster, C. gigas. The publication of the complete genome in 2012 has opened up new possibilities for the study of innate immunity in this species. The study of autophagy for the first time in oysters consisted in a first step in the in silico search for genes involved in this pathway and the corresponding proteins by Western blotting. Then, the role of this important process in innate immunity has been explored through reproduction infections tests with or without modulators of autophagy. Results showed that autophagy appeared as involved in defence mechanisms against viral and bacterial infection in Pacific oysters
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Allen, Sarah Kathryn. "Flow Cytometric Analysis of Crayfish Hemocytes". Digital Commons @ East Tennessee State University, 2011. https://dc.etsu.edu/etd/1309.
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Crayfish exhibit innate immune responses via hemocytes and their products. There are 3 hemocyte populations: hyaline cells, granular cells, and semigranular cells. Hemocytes from laboratory housed, untreated crayfish (normal crayfish) have been quantified on the basis of cell type, cell size, and cell granularity using Flow Cytometry. These data present the first overall picture of normal hemocytes from Red Swamp Crayfish with regard to cell type, cell size, and cell granularity and will serve as a baseline for all future studies in our lab. Experiments using crayfish injected with Pseudomonas aeruginosa, Staphylococcus aureus, or crayfish saline alone showed significant and consistent changes in cell type in cells from crayfish injected with bacteria with a decrease in hyaline cells and an increase in granular cells. This effect was greater in crayfish injected with Gram - bacteria. In addition, crayfish injected with Pseudomonas aeruginosa showed a significant difference in Granular cell size with a shift to larger cells and a significant decrease in granularity in the Granular cell population. Cells from crayfish treated with Staphylococcus aureus did not show these changes.
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Delmas, Jean-Claude. "Adaptation parasitaire de paecilomyces fumosoroseus (wize) brown et smith a l'insecte pieris brassicae l. (lep. Pieridae) et consequences hematologiques de l'infection". Paris 7, 1988. http://www.theses.fr/1988PA077048.
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Analyse comparative du comportement de 2 souches de p. F. De pathogenicite differente. L'etude clef du pouvoir entomopathogene a lieu au niveau de la reconnaissance de la cuticule par l'apex du tube germinatif. Il existe des sites preferentiels tegumentaires d'infection. Le genre pathogene est reconnu par les hemocytes, mais l'activite cytotoxique du champignon empeche la formation de granulone. La contamination de blessures hemorragiques par l'isolat non pathogene entraine une cicatrisation par les hemocytes qui peut bloquer l'infection fongique. L'analyse de l'ultrastructure des hemocytes des insectes sains conduit a identifier divers types cellulaires de p. B. Et a elaborer une hypothese sur le role des inclusions cytoplasmiques chez les hemocytes granuleux
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Rush, Craig Michael. "Crosstalk between the Jak-Stat and Wingless pathways is mediated by Mad in Drosophila melanogaster larval hematopoiesis". Ohio University Art and Sciences Honors Theses / OhioLINK, 2013. http://rave.ohiolink.edu/etdc/view?acc_num=ouashonors1367508013.
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Tucker, Philippa. "A genetic dissection of actin regulation in Drosophila hemocytes". Thesis, University of Bath, 2011. https://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.557798.
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Cell migration is essential for embryonic development, it occurs in adult organisms during processes like wound healing and its misregulation contributes to pathological conditions such as metastasis. Despite this, most studies of cell migration have been undertaken in vitro. Ena/VASP proteins, believed to be actin anti-capping proteins, have been studied extensively in fibroblasts in vitro, and using Drosophila macrophages (hemocytes) within the developing embryo, the role of the Drosophila homologue of Mena, Ena, is investigated in vivo. Consistent with data from fibroblasts in vitro, Ena localised to regions of actin dynamics within migratory hemocytes, where this protein stimulated lamellipodial dynamics and positively regulated filopodial number and length. However, whilst overexpression of Ena/VASP proteins in fibroblasts reduced migration speeds, Ena overexpression in hemocytes dramatically increased migration speeds in three different assays. This positive regulation of migration speed closely resembled the increased motility of breast cancer cells that overexpress Mena and evidence presented here, suggests that this key difference may be explained by spatial constraints that are imposed upon cells within three dimensional environments. Indeed, such constraints prevented ruffling, a more detrimental form of retraction, in hemocytes in vivo. Furthermore, fibroblasts overexpressing Mena in vitro form membrane ruffles more frequently. Therefore Ena/VASP proteins drive migration by enhancing lamellipodial protrusion, but in certain environments these protrusions are lost as ruffles slowing migration. The method by which Ena regulates lamellipodial protrusion and migration speeds was then investigated: Ena increased Fascin-mediated actin bundling and the number of Fascin rich-actin bundles that coalesced. Analysis of individual actin bundles revealed that coalescence increased protrusion rate and that both protrusion rate and coalescence, increased cell migration speeds. This suggests that Ena facilitates an increase in cell migration by promoting the coalescence of Fascin bundles, and positions Ena as a key regulator of migration speeds in vivo.
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Jönsson, Anders, e Thomas Nilsson. "HemoCue Cognatus - Ett Intelligent Analysinstrument". Thesis, Halmstad University, School of Business and Engineering (SET), 2008. http://urn.kb.se/resolve?urn=urn:nbn:se:hh:diva-1610.
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HemoCue AB is a multinational corporation targeted on manufacturing point-of-care analysis
instruments for medical treatment. Their main product, the DM 201, is outdated and has big
issues gaining new market shares. For example, no medical treatment center in Sweden uses
DM 201 frequently. The project of developing a new instrument to replace it was assigned
two students at the development engineering program at Halmstad University. The project
also included integration of the new platform for DM 202 to HemoCue AB:s remaining
products.
The project began by defining the different technical areas concerned. By identifying six big
technical areas and reaching a technical height within these areas the project group was able
to combine the new knowledge and assemble them to a satisfying result. The project group
developed a new instrument with a 50 % decreased volume compared to the DM 201. The
components are carefully selected to match the technical progress in the medical treatment
business over the past years. The original project also resulted in a side project containing a
new cuvette unit which is decreased by 75 % compared with today’s unit. This new cuvette
unit also performs more accurate analysis. When the goals where reached for DM 202, the
project group started to integrate useful components into the remaining product range. To
make this possible the group had to redesign all the remaining instruments both internal and
external.
The superior work of the project group has resulted in a completely new product range for
HemoCue AB. Consisting of intelligent, powerful instrument that will be very attractive on
the market.
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Lundberg, Torgny. "Betydelsen för DNA-metylering för differentiering av hemocyter i sötvattenskräftan Pacifastacus leniusculus". Thesis, Uppsala universitet, Institutionen för biologisk grundutbildning, 2021. http://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-433078.
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Reis, L?gia Garcia. "Efeito da salinidade, densidade de estocagem e da infec??o hipodermal e necrose hematopoi?tica (IHHN) na imunidade do camar?o Litopenaeus vannamei cultivados em fazendas do Rio Grande do Norte". Universidade Federal do Rio Grande do Norte, 2008. http://repositorio.ufrn.br:8080/jspui/handle/123456789/13044.
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Previous issue date: 2008-08-29The main problem faced by the shrimp industry are the infectious diseases. The hypodermal and hematopoietic necrosis infection (IHHN) is one of the major cause of disease in the cultured shrimp, Litopenaeus vannamei. Environmental changes involving water quality, oxygen concentration, salinity, temperature, stocking density, presence of pathogens, among others, triggering a stressing condition for the cultured shrimp, weakening them and allowing the outbreak of diseases. The stress on the animal leads to a change in the molecules immune response components, which can be used as indicators of shrimp health. Thus, the objective of the present study was to evaluate the effect of salinity, stocking density and IHHNV infection on the L. vannamei shrimp. The immune parameters used to check the shrimp health were the total hemocytes counts (THC), the agglutinating activity (AA) and the clotting time (CT) of the serum of shrimp. These parameters were analyzed in healthy and IHHNV-infected shrimp, grown in low (0-0.5 ), medium (19-24 ) and high (> 38 ) salinity, and extensive (7-12 cam.m-2), semi-intensive (15-25 cam.m-2) and intensive (33-45 cam.m -2) stocking density. The IHHNV infection rate was significantly higher in low salinity (P<0.005) and intensive density (P<0.005), both stressful conditions for L. vannamei. Low salinity significantly increased THC (P<0.05) and decreased and CT (P<0.05) in healthy and infected shrimp, but AA (P<0.05) significantly decreased in healthy shrimp at medium salinity. Culture intensification did not affect the THC, AA and CT of healthy and infected shrimp (P>0.05). The IHHNV infection did not affect any immune parameters of shrimp cultured at different salinities and stocking densities. It is necessary to emphasize that this study was conducted in shrimp grown in ponds, where several environmental factors are acting simultaneously. Thus, further studies are needed about the influence of other environmental factors on the immune parameters of shrimp cultured in pond
O principal problema enfrentado pela ind?stria do camar?o s?o as enfermidades de origem infecciosa. A Infec??o Hipodermal e Necrose Hematopoi?tica (IHHN) ? uma das principais causas de doen?as no camar?o de cultivo Litopenaeus vannamei. Altera??es do ambiente de cultivo envolvendo qualidade da ?gua, concentra??o de oxig?nio, salinidade, temperatura, densidade de estocagem, presen?a de pat?genos, entre outros, desencadeiam uma situa??o de estresse nos camar?es cultivados, debilitando-os e permitindo a instala??o de enfermidades. O estresse desencadeia no animal a altera??o de mol?culas componentes da resposta imune, que podem ser usadas como indicadores de sa?de do camar?o. Desta forma, o objetivo do presente trabalho foi avaliar o efeito da salinidade, densidade de estocagem e infec??o pelo IHHNV na imunidade do camar?o de cultivo L. vannamei. Os par?metros imunes utilizados para monitorar as condi??es de sa?de dos camar?es foram o n?mero total de hem?citos (THC), a atividade aglutinante (AA) e o tempo de coagula??o (TC) dos soros dos camar?es. Estes par?metros foram analisados em camar?es saud?veis e infectados pelo IHHNV, cultivados em baixa (0-0,5 ), m?dia (19-24 ) e alta (>38 ) salinidades e em extensiva (7-12 cam.m-2 ), semi-intensiva (15-25 cam.m-2) e intensiva (33-45 cam.m-2) densidades de estocagem. A taxa de infec??o pelo IHHNV foi significativamente maior em baixa salinidade (P<0,005) e na densidade intensiva (P<0,005), ambas condi??es estressantes para o L. vannamei. Baixa salinidade significativamente aumentou THC (P<0,05) e diminuiu TC (P<0,05) de camar?es saud?veis e infectados, mas AA (P<0,05) de camar?es saud?veis diminui significativamente na m?dia salinidade. A intensifica??o do cultivo n?o afetou THC, AA e TC de camar?es saud?veis e infectados (P>0,05). A infec??o pelo IHHNV n?o afetou nenhum dos par?metros imunes dos camar?es cultivados nas diferentes salinidades e densidades de estocagem. ? necess?rio enfatizar que o presente estudo foi realizado em camar?es cultivados em viveiros, onde v?rios fatores ambientais est?o atuando simultaneamente. Desta forma, s?o necess?rios estudos adicionais sobre a influ?ncia de outros fatores ambientais nos par?metros imunes de camar?es nas condi??es de cultivo
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Wong, Christine. "Regulation of inflammation genetic control of the transcription of Spätzle in Drosophila hemocytes". Thesis, King's College London (University of London), 2013. https://kclpure.kcl.ac.uk/portal/en/theses/regulation-of-inflammation-genetic-control-of-the-transcription-of-spatzle-in-drosophila-hemocytes(a29adfcc-f504-4a7a-8a1c-db74fe1a20df).html.
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The Drosophila cytokine Spätzle (Spz) triggers the Toll signalling cascade in flies upon infection with Gram-‐positive bacteria and fungi. It is an analogue to the mammalian IL-‐1 and is produced by the fat body as a precursor protein (Spz-‐ precursor). Active Spz is produced from its precursor by a tightly regulated cascade of serine proteases. However, the transcriptional regulation of Spz in Drosophila in response to bacterial or fungal infection is still poorly deciphered. Therefore, the aim of the project was to interrogate the regulation of Spz transcription, with specific interest in hemocytes, which are equivalent to mammalian monocytes/macrophages. For this purpose, we have generated and characterised a transgenic Spz-‐neGFP reporter that allows the visualisation of Spz transcription in individual cells and tissues during different stages of development. We have identified populations of cells that transcribe the Spz gene in the steady state throughout development. We have infected the transgenic reporter flies with the fungus Candida albicans and examined the transcription of Spz after immune challenge. In future, these strains and results will help develop an in vivo genome wide assay, employing Drosophila RNAi libraries, in order to gain more knowledge about the transcriptional regulation of Spz and to identify novel regulatory genes and pathways for Spz transcription.
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Price, Regan R. "Clic Modulates Filopodia Formation Downstream of Cdc42 and its Effectors in Drosophila Hemocytes". Ohio University Honors Tutorial College / OhioLINK, 2012. http://rave.ohiolink.edu/etdc/view?acc_num=ouhonors1338571019.
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BRAUN, EGLES ANNE. "Etude des hemocytes de drosophile et de leur role dans la reponse immunitaire". Université Louis Pasteur (Strasbourg) (1971-2008), 1998. http://www.theses.fr/1998STR13047.
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L'objectif de ce travail de these etait d'etudier les cellules sanguines (ou hemocytes) de drosophile et leur role au cours de la reponse immunitaire. Dans un premier temps, nous avons recherche de nouveaux outils pour l'etude des hemocytes au stade larvaire. Pour ceci, nous avons utilise la technique de l'enhancer trap. Cette technique se base sur l'insertion dans le genome, d'un transposon contenant un gene rapporteur place sous le controle d'un promoteur minimal et ubiquitaire. Le transposon s'insere frequemment dans les regions regulatrices des genes et se place ainsi sous leur controle. Dans ce cas, l'expression du transposon reflete celle du gene hote et peut etre suivie grace au gene rapporteur. De plus, une telle insertion genere souvent une mutation. Nous avons crible 1341 lignees enhancer trap de drosophile en recherchant d'une part, une expression du gene rapporteur dans les hemocytes larvaires, et d'autre part, des phenotypes en relation avec les cellules sanguines. Nous avons ainsi identifie plusieurs lignees correspondant a des marqueurs d'hemocytes larvaires. Nous avons egalement isole un nouveau mutant caracterise par l'absence d'hemocytes larvaires et par une mort cellulaire importante dans l'organe hematopoietiques ; ce mutant a ete appele domino. Dans un second temps, grace a domino, nous avons etudie le role des hemocytes dans la reponse immunitaire chez la larve de drosophile. Nous avons montre que : (i) les hemocytes ne sont pas necessaires pour l'induction de la synthese des peptides antimicrobiens en reponse a une blessure septique ; (ii) en absence d'hemocytes, les reactions de melanisation sont affectees, mais pas abolies. Des experiences de survies larvaires suite a une blessure ou a une infection revelent l'importance des cellules sanguines dans les processus de defense de l'hote. Enfin, la troisieme partie de cette these a consiste en une etude fonctionnelle et moleculaire de domino. Nos resultats indiquent que domino est implique dans la proliferation cellulaire, qu'il code pour deux transcrits de 10 et 11 kb et que la proteine correspondante contient un domaine atpase adn-dependant, et appartient de ce fait a la famille des proteines swi2/snf2.
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Tincu, John Andrew. "Acid soluble peptides from the hemocytes of the ascidians : characterization and antimicrobial activity /". Diss., Connect to a 24 p. preview or request complete full text in PDF format. Access restricted to UC campuses, 2003. http://wwwlib.umi.com/cr/ucsd/fullcit?p3118434.
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Barrett, Andrew Paul. "Investigating the roles of CNS-patterning molecules and integrins during midline migration of Drosophila hemocytes". Thesis, University of Kent, 2007. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.445716.
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Feldt, Olivia. "Evaluation of underfill-function in HemoCue Monitor, a POCT-instrument". Thesis, Uppsala University, Department of Medical Biochemistry and Microbiology, 2006. http://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-6991.
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Objective: The aim of this study was to evaluate a new underfill-function in a POCT-instrument from HemoCue AB (Ängelholm, Sweden). The instrument is in use today among diabetes patients for self-monitoring blood glucose (SMBG). The new function is supposed to guarantee that measuring only will be performed on a sufficient sample volume to assure that the correct glucose value is received.
Methods and results: Blood samples (whole blood) from 12 patients were analysed with the instrument. Measuring were performed using different volumes in the cuvette. Full cuvette, 3µL, 2µL, 1µL and a measuring on an empty cuvette. The instrument performed measurements on all volumes added to the cuvette except for the empty cuvette. The less sample volume that was used the lower glucose values were reported by the instrument.
Conclusions: The new under fill-function did not work satisfactory. If such function would be more reliable it would be beneficial for the patient controlling hers/his bloodglucose provided that the testing procedure is being correctly done. This is very important because the results are often used to treat the patient.
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Araújo, Helena Rocha Côrrea de. "Ultra-estrutura dos hemócitos de Aedes aegypti (Linnaeus, 1762) (Diptera, Culicidae)". Centro de Pesquisas Aggeu Magalhães, 2009. http://beta.arca.fiocruz.br/handle/icict/14543.
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Previous issue date: 2009Made available in DSpace on 2016-07-05T22:16:55Z (GMT). No. of bitstreams: 3 985.pdf.txt: 141267 bytes, checksum: a925e4eaf511d9977eb6efbe2cc45af4 (MD5) license.txt: 1748 bytes, checksum: 8a4605be74aa9ea9d79846c1fba20a33 (MD5) 985.pdf: 1940501 bytes, checksum: 018fcc4969f375effb75a13c7d6c7974 (MD5) Previous issue date: 2009
Fundação Oswaldo Cruz. Centro de Pesquisas Aggeu Magalhães. Recife, PE, Brasil
O sucesso dos insetos em explorar diversos ambientes é devido, em grande parte, à habilidade em se defender contra patógenos e parasitas. Nos insetos, os principais mecanismos de defesa são desempenhados pelos hemócitos. A classificação dos tipos de hemócitos presentes na hemolinfa ainda é bastante controversa. A biodiversidade desses organismos tem proporcionado modelos importantes para o estudo de suas estratégias antimicrobianas, as quais podem fornecer informações relevantes para o combate a diversas doenças, bem como para o estudo da imunologia geral. O objetivo do presente estudo foi caracterizar a resposta imune celular de Aedes aegypti sob o ponto de vista morfológico, funcional e ultra-estrutural através da microscopia óptica e eletrônica de transmissão. Em nossos estudos identificamos seis tipos morfológicos de hemócitos circulantes na hemolinfa de Ae. aegypti através da microscopia de luz, eletrônica de transmissão e contraste de interferência diferencial, são eles: prohemócitos, adipohemócitos, granulócitos, plasmatócitos, oenocitóides e trombocitóides. Os prohemócitos foram as menores células encontradas na hemolinfa. Sua principal característica é a presença de um citoplasma ocupando uma pequena área em torno do núcleo. Os adipohemócitos foram os mais abundantes tipos celulares presentes e exibiam grandes inclusões lipídicas preenchendo praticamente todo o citoplasma. Os granulócitos possuem um citoplasma contendo diversos grânulos elétron-densos. Os plasmatócitos exibiram morfologia bastante polimórfica e diversos filopódios e pseudópodes. Os oenocitóides possuem citoplasma homogêneo com poucas organelas. Os trombocitóides são raros e possuem características similares aos oenocitóides com organelas pouco desenvolvidas. Os hemócitos responsáveis pela resposta imune contra partículas de látex conjugadas a FITC foram identificados através da microscopia laser confocal, assim como as lectinas que marcam os hemócitos. Os granulócitos foram as únicas células envolvidas na fagocitose de alvos abióticos in vitro e in vivo. As lectinas BSI, ConA, HPA, LCA, PNA, UEA e WGA marcaram os hemócitos com variações na intensidade. A WFA e LPL não marcaram hemócitos de Ae. aegypti (AU).
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Dobrescu, Gelu. "Mannose and Lipopolysaccharide Receptors on the Surface of Granular Hemocytes from the Crayfish Procambarus clarkii". [Johnson City, Tenn. : East Tennessee State University], 2002. http://etd-submit.etsu.edu/etd/theses/available/etd-0307102-133116/unrestricted/dobrescug032602.pdf.
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Pyriohou, Anastasis. "Regeneration of the neural complex in the ascidian Ciona intestinalis with particular reference to the role of hemocytes". Thesis, Royal Holloway, University of London, 1999. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.298362.
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Edgren, Beatrice. "Effekt på leukocyter efter ett långvarigt fysiskt arbete : En studie gjord under Vasaloppet". Thesis, Linnéuniversitetet, Institutionen för idrottsvetenskap (ID), 2015. http://urn.kb.se/resolve?urn=urn:nbn:se:lnu:diva-43804.
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BAKGRUND: Leukocyter ingår i kroppens immunförsvar vilket skyddar oss mot b.la. angrepp av bakterier och virus. Vid högintensivt fysiskt arbete så hamnar vi i ett s.k. ”open window” där vi är som mest mottagliga för sjukdomar. Tidigare studier har visat att vid långvarig, högintensivt fysiskt arbete ökar leukocytantalet, vilket leder till att kroppen sätter upp ett försvar mot angrepp.SYFTE: Studiens syfte var att undersöka effekten på leukocyter och dess undergrupper, om de ökade efter ett långvarigt högintensivt fysiskt arbete, i detta fall Vasaloppet. METOD: Mätningar har gjorts via kapillärt blodprov, de gjordes innan start och efter målgång och analyserades sedan i Hemocue WBC Diff. RESULTAT: Totala leukocytantalet ökade med 6,8, neutrofila granulocyterna ökade med 6,7, monocyterna ökade med 0,2 och de basofila granulocyterna ökade med 0,03. Lymfocyterna minskade med -0,1 och de eosinofila granulocyterna minskade med -0,55. KONKLUSION: Långvarigt, ansträngande, fysiskt arbete, leder till en ökning av leukocyter och vissa undergrupper. Detta innebär att immunförsvaret påverkas och man hamnar i det så kallade ”open window”. Till framtida studier skulle det vara intressant att se hur immunförsvaret påverkas av träning under en längre period.
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Goins, Kimberly R. "Host Defense Mechanisms in the Crayfish: the Effect of Injection with Live or Killed Bacteria". [Johnson City, Tenn. : East Tennessee State University], 2003. http://etd-submit.etsu.edu/etd/theses/available/etd-0328103-141532/unrestricted/Goins04162003f.pdf.
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Thesis (M.S.)--East Tennessee State University, 2003.Title from electronic submission form. ETSU ETD database URN: etd-0328103-141532. Includes bibliographical references. Also available via Internet at the UMI web site.
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Evariste, Lauris. "Caractérisations structurale et fonctionnelle des populations hémocytaires de la moule zébrée (Dreissena sp.) en vue de leur utilisation en évaluation du risque écotoxicologique". Thesis, Reims, 2016. http://www.theses.fr/2016REIMS016/document.
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L’extension des activités humaines est responsable du rejet de molécules et de perturbations climatiques pouvant affecter la physiologie des organismes aquatiques. La moule zébrée possède des caractéristiques biologiques faisant d’elle une espèce intéressante en surveillance environnementale. Chez cet organisme, les hémocytes constituent une cible privilégiée pour la mise en place d’une approche multi-biomarqueurs. En effet, ces cellules à fonctionnalités multiples sont impliquées dans les grandes fonctions physiologiques de l’espèce et la régulation de l’homéostasie des individus. L’objectif de ce travail est de développer les outils analytiques permettant d’étudier les réponses hémocytaires de la moule zébrée. Les expérimentations menées ont permis de caractériser la structure des populations hémocytaires ainsi que leurs fonctionnalités propres en lien avec le processus de phagocytose. L’utilisation de ces biomarqueurs dans divers contextes indique une forte adaptabilité de l’espèce aux conditions environnementales. Les résultats montrent l’intérêt d’analyser les activités hémocytaires à l’échelle des sous populations comparativement à l’approche globale ne tenant pas compte de la diversité cellulaire. Il a été observé que certains facteurs comme le statut reproducteur ou l’espèce échantillonnée (D. polymorpha vs D. bugensis) constituent des facteurs de confusion importants. Il ressort également un positionnement fort du test de phagocytose en tant que marqueur de sensibilité aux contaminants. Ce travail constitue un ensemble de données voué à être utilisé dans des contextes multiples aussi bien en écotoxicologie qu’en écophysiologieExtension of human activities is responsible of molecule releases and climate changes that may affect physiology of aquatic organisms. The zebra mussel has biological traits making it an interesting species for environmental monitoring. In this organism, hemocyte cells constitute an interesting target to develop a multi-biomarker approach. These cells possess multiple functionalities and are involved in all major physiological functions of the species and in homeostasis regulation. The objective of this work was to develop analytical tools to study hemocyte responses of zebra mussels. Experiments allowed characterizing structure of hemocyte populations and their functionalities linked with phagocytosis process. Use of these biomarkers in various contexts indicated an important adaptation capacity of the species to environmental conditions. Results highlighted interest to analyze hemocyte activities at sub-population scale comparatively to global approach that does not consider hemocyte diversity. It was demonstrated that factor such as reproductive status or sampled species (D. polymorpha vs D. bugensis) constitute important confounding factors. Studies also demonstrated a strong positioning of phagocytosis assay as a sensitive marker to contaminants. This work constitutes a data set destined to be used in multiple contexts such as ecotoxicology or ecophysiology
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Carlsson, Nathalie. "Metodjämförelse mellan Hemocue WBC Diff, Advia 2120 och manuell mikroskopi avseende differentialräkning av leukocyter". Thesis, Linnéuniversitetet, Institutionen för naturvetenskap, NV, 2011. http://urn.kb.se/resolve?urn=urn:nbn:se:lnu:diva-13990.
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Patientnära instrument kan i vissa vårdsituationer ge snabba analysresultat på ett kostnadseffektivt sätt. En viktig patientnära analys är differentialräkning av leukocyter, som visar fördelning och utmognad av dessa blodceller. Störningar i bildningen av leukocyter sker vid infektioner och hematologiska sjukdomar. Med manuell mikroskopi räknas leukocyter i ett blodutstryk färgat med May-Grünwald Giemsa. En kvantitativ och kvalitativ bedömning utförs av alla blodceller. Advia 2120 utför differentialräkning av leukocyter genom cytokemiska reaktioner och ljusspridning i olika vinklar vid belysning av celler. Celler identifieras baserat på kärnkonfiguration, granulering och cellstorlek.Hemocue White Blood Cell (WBC) Diff har en charged coupled device (CCD) kamera som scannar celler i en mikrokyvett. Genom beräkning av optisk densitet från pixlars gråskalor och jämförelse med referensceller identifieras celler. Syftet med studien var att jämföra analysresultat gällande differentialräkning av leukocyter i venöst blod mellan Hemocue WBC Diff och Advia 2120 samt i kapillärt blod mellan Hemocue WBC Diff och manuell mikroskopi. Provmaterialet bestod av 62 venösa prover från patienter och 20 kapillära prover från friska vuxna personer. Resultatet vid jämförelse mellan Hemocue WBC Diff och Advia 2120 visade en korrelation på 0,99 vid beräkning av totala antalet leukocyter, neutrofila granulocyter och lymfocyter. En större spridning av resultat sågs vid beräkning av monocyter, eosinofila- och basofila granulocyter. Resultatet vid jämförelse mellan Hemocue WBC Diff och manuell mikroskopi visade en korrelation på 0,95 för neutrofila granulocyter och 0,84 för lymfocyter. För de andra celltyperna var korrelationen dålig.Hemocue WBC Diff fungerar bra vid beräkning av leukocyter på friska personer. Instrumentet är dock under utveckling och förbättringar på vissa parametrar kan behövas.
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Andersson, Sebastian. "Lipemi-interferens vid mätning av Hb på Sysmex XN-10 och HemoCue Hb 201+". Thesis, Högskolan Kristianstad, Fakulteten för naturvetenskap, 2019. http://urn.kb.se/resolve?urn=urn:nbn:se:hkr:diva-19225.
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Anemi kan uppstå till följd av förlust av erytrocyter eller försämrad produktion av nya erytrocyter. För att upptäcka och följa upp patienter med anemi är det viktigt att korrekt kunna mäta hemoglobinkoncentrationen (Hb) i blodet. En vanlig metod för att mäta Hb-koncentration är fotometri i kombination med en kemisk omvandling. Liksom alla mätningar med ljus är dessa känsliga för turbiditet i provet. Lipemi är en vanlig källa till turbiditet som kan uppstå till exempel som följd av en fettrik måltid, diabetes mellitus, lever- eller njursjukdomar, alkoholism och vissa läkemedel. Olika instrumenttillverkare har olika metoder för att motverka interferensen av lipemi. Sysmex hematologi-instrument XN-10 använder en fettlösande bärarvätska i sin fotometriska kanal (HGB) och HemoCue mäter vid en andra våglängd som ska kompensera för turbiditet. Sysmex XN-10 har också en optisk kanal (HGB-O) som är till för att räkna retikulocyter genom att mäta deras nukleinsyra- samt Hb-innehåll men ger då också ett beräknat värde på Hb-koncentrationen i hela provet. Syftet med denna studie var att jämföra HGB och den HGB-O för bestämning av Hb-koncentrationen i helblod. Båda kanalerna jämfördes även med HemoCue Hb 201+ vid bestämning av Hbkoncentrationen i svårt lipemiska prover. Hb-mätning på plasma från motsvarande prover utfördes också för att undersöka om värdet motsvarade Hb-höjningen i de lipemiska proverna. Prover analyserade med både HGB och HGB-O på Sysmex XN-10 på klinisk kemi vid Skånes universitetssjukhus i Lund under november månad 2018 (n = 392) jämfördes med hjälp av Spearmans rangkorrelationskoefficient. Lipemi simulerades med fettemulsionen Intralipid i totalt 32 prover. Färdiganalyserade patientprover från föregående dag delades i ett lipemiskt prov med Intralipidtillsats och ett nollprov med tillsats av NaCl-lösning i en motsvarande volym. Differenserna mellan de lipemiska- och nollprovernas Hb-värden testades för signifikans med icke-parametrisk Wilcoxons teckenrangtest. Kruskal-Wallis samt Dunns's tester användes för att visa på signifikanta skillnader mellan de tre metoderna. Signifikansnivån sattes vid p < 0,05. Resultaten visade god korrelation mellan HGB - och HGB-O Hb-värden med ett Spearman korrelationsvärde på 0,982. Jämförelsen av metoderna vid lipemi visade signifikant skillnad mellan nollprov och lipemiskt prov för HGB- (p < 0,001) men inte HGB-O (p = 0,11) på XN-10. HemoCue Hb 201+ visade också signifikant skillnad (p < 0,001) vid lipemi men med lägre median-värde än HGB och mindre spridning än HGB-O. HGB-O:s median-värde tydde på minst lipemipåverkan men spridningen av differenserna var stor. Spridningen av HGB-O resultaten kan bero på hemolys då endast intracellulärt Hb mäts i denna kanal. Resultaten i denna studie tyder på att HemoCue-metoden är den mest pålitliga vid Hb-mätning av lipemiska prover och därmed det lämpligaste komplementet till HGBmetodenAnemia can arise from either loss of erythrocytes or impaired production of new erythrocytes. In order to discover and evaluate the treatment of anemic patients, correct Hb measurements are important. A common method to measure Hb concentration is photometry in combination with chemical conversion of the Hb. Like all light-dependent methods this suffers from a vulnerability to turbidity that scatters light. Lipemia is a common cause of turbidity caused by e.g. recent intake of high fat foods, diabetes mellitus, liver or kidney disease, alcoholism and some drugs. Manufacturers of Hb analyzers use different methods to counter the influence of interference from lipemia on measurements. Sysmex XN-10 analyzers use a fat dissolving sheath fluid in its photometric channel (HGB) and HemoCue measures absorbance at a second wavelength to compensate for turbidity. Sysmex XN-10 also has an optic channel (HGB-O) for counting reticulocytes by measuring their nucleic acid and Hb content. At the same time this channel measures Hb equivalents of erythrocytes and gives a calculated value of Hb content in the entire sample. The aim of this study was to compare the photometric and the optical channels for measuring Hb concentration in whole blood. Both the Sysmex XN-10 channels were compared with HemoCue Hb 201+ when measuring Hb concentrations in lipemic samples. Plasma Hb concentration was determined for the corresponding samples in order to investigate correlation between elevation in Hb concentration with and without simulated lipemia and in the plasma after centrifugation. Samples analyzed at Skånes University Hospital in Lund during the month of November 2018 (n = 392) using both HGB and HGB-O on XN-10 were compared using Spearman's signed correlations coefficient. Lipemia was simulated by using the fat emulsion Intralipid in a total of 32 samples. Samples collected and analyzed on the previous day was used for the study. Each sample was split into one part with added Intralipid to form a lipemic sample and one part with NaCl-solution of the same volume as Intralipid in the lipemic sample. The differences between lipemic and non lipemic samples was tested for significance by the non-parametric Wilcoxons signed ranks test for each of the methods. Significance between the three methods was tested by using Kruskal-Wallis and Dunn's tests. Level of significance was set to p < 0.05. The results showed good correlation between earlier test run on both HGB and HGB-O with a Spearman correlation score of 0,982. A significant difference was found between lipemic and non lipemic samples with the photometric method (p < 0,001) but not the optical method (p = 0,11) on XN10. HemoCue Hb 201+ also showed a significant difference (p < 0,001) between lipemic and non lipemic samples but a lower median than HGB and less deviation than HGB-O. The median of HGB-O indicated that it was influenced the least by lipemia of the three methods but had the greatest deviation of the differences. The greater deviation of HGB-O values may have been caused by hemolysis since the method measures intra cellular Hb. HemoCue shows according to this study the slightest deviation of the three methods and a less heightened median value compared to HGB which confirms the methods suitability as complement to HGB when dealing with lipemic samples.
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Fukuzawa, Aline Harumi. "Envolvimento dos hemócitos na resposta imune da aranha caranguejeira Acanthoscurria gomesiana". Universidade de São Paulo, 2007. http://www.teses.usp.br/teses/disponiveis/42/42135/tde-30012008-115755/.
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Os invertebrados impedem o estabelecimento de uma infecção através de uma resposta imune eficiente. Esta resposta envolve reações celulares e humorais. Existem poucos trabalhos sobre a imunidade das aranhas. O principal objetivo deste estudo foi verificar o papel dos hemócitos e dos peptídeos antimicrobianos na resposta imune da aranha Acanthoscurria gomesiana. Inicialmente, a localização relativa da gomesina e da acanthoscurrina foi determinada, mostrando que 58% dos hemócitos armazenam os dois peptídeos antimicrobianos. Além disso, foi verificado que a gomesina é direcionada aos grânulos dos hemócitos da forma de pró-peptídeo. Observou-se ainda, que após um desafio, os hemócitos migram para o sítio de infecção, onde deve secretar componentes da cascata de coagulação e peptídeos antimicrobianos. Além disso, os resultados mostraram que a fagocitose não é o principal mecanismo ativado após uma infecção. Assim sendo, as principais respostas imunes da aranha são através da coagulação e secreção dos peptídeos antimicrobianos.Invertebrates avoid the infection establishment through an efficient immune response. This response consists in cellular and humoral reactions. Few data are available concerning the spider\'s immunity. The main aim of this study was to determine the role of hemocytes and antimicrobial peptides on the immunity of the spider Acanthoscurria gomesiana. Initially, the localization of gomesin and acanthoscurrin was determined, showing that 58% of hemocytes store both antimicrobial peptides. Moreover, our results show that gomesin is addressed to the hemocyte granules as a pro-peptide. We also demonstrate, by in vivo and in vitro experiments, that hemocytes migrate to the site of microbial infection. Once at the site of infection, hemocytes might secrete components of coagulation cascade and antimicrobial peptides. Besides, our results suggest that phagocytosis is not the major defense mechanism activated after microbial challenge. Therefore the main reactions involved in the spider defense might be through the coagulation and antimicrobial peptides secretion.
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Gustavsson, Frida. "Validation study : HemoCue Hb 201 + as a tool in comparative physiological field studies on avian blood". Thesis, Linköpings universitet, Biologi, 2015. http://urn.kb.se/resolve?urn=urn:nbn:se:liu:diva-120106.
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Haemoglobin concentration is becoming a widely popular parameter to use to assess physiological condition within a broad range of species. Assessments of large populations would preferable be done in field to receive quick results and avoid confounding factors associated with transport of blood. A validation study is here performed to see how well the point-of-care device HemoCue Hb 201 + can assess haemoglobin concentration on avian blood. Nucleated erythrocytes have previously been pointed out as something that makes it problematic to apply HemoCue Hb 201 +, designed for human blood, on avian blood. Here it is shown that HemoCue Hb 201 + accurately can estimate haemoglobin concentration for chicken-, tinamou-, and ostrich blood. However, manipulation of ostrich cells, to yield a larger mean corspuscular volume, results in HemoCue Hb 201 + overestimating haemoglobin concentration. A large mean corpuscular volume could therefore be something that impair accuracy in values retrieved with HemoCue Hb 201 +. This study shows that HemoCue Hb 201 + seems possible to apply on avian blood to some extent, but highlights the importance of validation studies when applying this device on new species.
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Diao, Yupu, Anrui Lu, Bing Yang, Wenli Hu, Qing Peng, Qing-Zhi Ling, Brenda T. Beerntsen, Kenneth Söderhäll e Erjun Ling. "Existence of Prophenoloxidase in Wing Discs : A Source of Plasma Prophenoloxidase in the Silkworm, Bombyx mori". Uppsala universitet, Jämförande fysiologi, 2012. http://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-184482.
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In insects, hemocytes are considered as the only source of plasma prophenoloxidase (PPO). PPO also exists in the hemocytes of the hematopoietic organ that is connected to the wing disc of Bombyx mori. It is unknown whether there are other cells or tissues that can produce PPO and release it into the hemolymph besides circulating hemocytes. In this study, we use the silkworm as a model to explore this possibility. Through tissue staining and biochemical assays, we found that wing discs contain PPO that can be released into the culture medium in vitro. An in situ assay showed that some cells in the cavity of wing discs have PPO1 and PPO2 mRNA. We conclude that the hematopoietic organ may wrongly release hemocytes into wing discs since they are connected through many tubes as repost in previous paper. In wing discs, the infiltrating hemocytes produce and release PPO probably through cell lysis and the PPO is later transported into hemolymph. Therefore, this might be another source of plasma PPO in the silkworm: some infiltrated hemocytes sourced from the hematopoietic organ release PPO via wing discs.
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Silva, Daniella Gonçalves da. "Efeito do fungo Trichoderma harzianum e do zinco em colônias de Atta sexdens". Universidade Federal do Tocantins, 2016. http://hdl.handle.net/11612/367.
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As formigas-cortadeiras do gênero Atta são apontadas como os principais herbívoros da região Neotropical, sendo capazes de causar grandes danos à agricultura, às pastagens e à silvicultura em especial. No controle químico dessa praga, a tática mais usual e efetiva tem sido o uso de iscas granuladas tóxicas. Todavia, têm-se procurado métodos alternativos para o controle de cortadeiras, sobretudo por pressão de agências certificadoras de manejo florestal como o FSC (Forest Standarship Council). Recentemente, isolados de Trichoderma spp. começaram a ser testados no controle de formigas-cortadeiras em razão das suas propriedades antagonísticas ao fungo simbionte por elas cultivados. Além disso, destacam-se substâncias que têm o potencial de inibir a resposta imune inata dos insetos. Por exemplo, alguns elementos químicos como, cádmio e zinco. O presente trabalho objetivou o preparo de uma formulação com iscas granuladas e encapsuladas do fungo Trichoderma harzianum. Para encapsulação do fungo, utilizou-se uma mistura de alginato de sódio, farelo de trigo, suco concentrado de laranja e micélio triturado do antagonista. Esta mistura foi gotejada em solução (0,25 M) de CaCl2, o que permitiu a formação de grânulos esféricos de diâmetro regular. Paralelamente testou-se iscas contendo sulfato de zinco ZnSO4 (0,25 g/L), produzidas a partir da mistura de alginato, farelo de trigo e suco concentrado de laranja. Após o fornecimento das iscas fez-se a contagem total de hemócitos das operárias a fim de verificar alterações da sua resposta imune. Não ocorreu declínio na quantidade de hemócitos. Apesar das iscas não terem promovido a morte das colônias, elas apresentaram boa aceitação pelas operárias e promoveram a redução do volume do fungo simbionte. O cloreto e o sulfato de zinco foram empregados nas concentrações de 0,15; 0,25; 0,5; 1,5; 2;5 e 5;0 g/L em placas de Petri em meio BDA para o teste de desenvolvimento dos fungos simbionte e antagonista, e os resultados mostraram inibição no crescimento nas doses máximas tanto em Leucoagaricus gongylophorus como em Trichoderma harzianum. As operárias foram imersas em soluções de sulfato de zinco com as mesmas concentrações daquelas empregadas no teste de inibição dos fungos. Após o tempo de 24 e 48 horas fez-se a contagem total de hemócitos e verificou-se um decréscimo dos mesmos em altas concentrações. Conclui-se que as iscas contendo T. harzianum e sulfato de zinco apresentaram boa aceitação por parte das colônias, elas não promoveram a morte das colônias, no entanto, reduziram o volume do fungo simbionte. Altas doses de cloreto e sulfato de zinco inibem o desenvolvimento do fungo antagonista e do fungo simbionte e elevadas concentrações zinco e o maior tempo de exposição das operárias ao mesmo afetam o seu sistema imune.The leaf-cutting ants of the genus Atta are cited as the main herbivores of the Neotropical region, being capable of causing major damage to agriculture, pasture and forestry in particular. In the chemical control of this plague, the most common and effective tactic has been the use of toxic granular baits. However, there have been alternative methods for control of cutting, especially by pressure certifying agencies forest management as the FSC (Forest Standarship Council). Recently, Trichoderma spp. They began to be tested in the control of leaf-cutting ants because of their antagonistic properties to the symbiotic fungus cultivated by them. Furthermore, they highlight substances that have the potential of inhibiting the innate immune response of the insects. For example, some chemical elements such as cadmium and zinc. This study aimed to the preparation of a formulation with granulated baits and encapsulated fungus Trichoderma harzianum. For encapsulation fungus, a mixture of sodium alginate was used wheat bran, concentrated orange juice and triturated antagonist mycelium. This mixture was dripped into solution (0,25 M) CaCl2, which allowed the formation of spherical granules of regular diameter. Parallel tested for baits containing zinc sulfate ZnSO4 (0,25 g/L) produced from the mixture of alginate, wheat bran and concentrated orange juice. After the supply of baits made up the total count of hemocytes of the workers in order to verify changes in their immune response. There was no decline in the amount of hemocytes. Despite the baits have not promoted the death of the colonies, they had good acceptance by workers and promoted the reduction of the symbiont fungus volume. Chloride and zinc sulfate were used in concentrations of 0.15; 0.25; 0.5; 1.5; 2, 5 and 5; 0 g /L in Petri dishes on PDA medium for the development and test of antagonist symbiont fungi, and the results showed growth inhibition in both maximal doses Leucoagaricus gongylophorus as Trichoderma harzianum. The ants were dipped in zinc sulfate solutions with the same concentrations of those employed in the fungal inhibition assay. After time 24 and 48 hours we made the total hemocytes count and there was a decrease in high concentrations thereof. We conclude that the baits containing T. harzianum and zinc sulfate showed good acceptance by the colonies, they did not promote the death of the colonies, however, reduced the volume of the symbiont fungus. High doses of zinc chloride and sulfate inhibit the development of the antagonist fungus and symbiont fungus and high concentrations of zinc and the longer exposure time of workers at the same affect your immune system.
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Risberg, Josefin. "Abbott Precision Xceed Pro har större mätosäkerhet än HemoCue Glucose 201+ : Utvärdering av en blodglukosmätare för patientnära analyser". Thesis, Umeå universitet, Biomedicinsk laboratorievetenskap, 2011. http://urn.kb.se/resolve?urn=urn:nbn:se:umu:diva-58645.
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Pereira, Lourivaldo dos Santos. "Resposta imune do carrapato bovino Boophilus microplus: investigação da produção de espécies reativas de oxigênio pelos hemócitos". Universidade de São Paulo, 2000. http://www.teses.usp.br/teses/disponiveis/42/42135/tde-28032001-105453/.
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Neste estudo avaliamos a ocorrência de fagocitose por parte de alguns tipos celulares presentes na hemolinfa do carrapato bovino B. microplus e a produção de espécies reativas de oxigênio durante a resposta imune. As técnicas empregadas para avaliação da produção de espécies reativas de oxigênio foram luminescência amplificada por luminol, oxidação de fenol vermelho, microscopia de fluorescência e fluorimetria com o corante dihidrorrodamina 123 (DHR). Observamos um aumento da luminescência amplificada por luminol quando hemócitos foram incubados na presença de bactérias Micrococcus luteus ou zimosam ou PMA. Esta luminescência foi inibida por superóxido dismutase (SOD) e por catalase (CAT), enzimas antioxidantes que removem superóxido e peróxido de hidrogênio, respectivamente. LPS não elicitou aumento da luminescência dos hemócitos em relação ao controle. Através da oxidação de fenol vermelho em reação inibida por CAT, verificamos aumento nos níveis de H2O2 produzido pelos hemócitos quando estimulados com PMA e Micrococcus luteus, enquanto não houve aumento quando o estímulo foi LPS, corroborando os resultados da luminescência. Usando microscopia de fluorescência para avaliar a produção de ERO pelos hemócitos, encontramos que cerca de 25% dos hemócitos fluorescem com maior intensidade quando estimulados com zimosam, sendo esta fluorescência inibida por CAT. Através de fluorimetria usando DHR observamos um aumento na intensidade de fluorescência dos hemócitos estimulados com PMA em reação inibida por cisteína, substância redutora que remove peróxido de hidrogênio e peroxinitrito. Nosso conjunto de resultados permitem concluir que os hemócitos do carrapato bovino produzem espécies reativas de oxigênio durante a resposta imune, semelhantemente ao que ocorre em vertebrados e em invertebrados como moluscos, crustáceos e insetos. Este é o primeiro trabalho mostrando produção de ERO pelos hemócitos de aracnídeos.The phagocytic activity and the reactive oxygen species (ROS) production during immune response of some hemocytes of the cattle tick Boophilus microplus were evaluated in this study. The ROS production was evaluated by luminol amplified luminescence, phenol red oxidation, dyhydrorhodamine (DHR) fluorescence microscopy and fluorimetry. The luminol-amplified luminescence increased when hemocytes were incubated with bacteria (Micrococcus luteus) or zymosam or phorbol 12-miristate 13 acetate (PMA). The superoxide dismutase (SOD) and catalase (CAT), antioxidant enzymes that removes superoxide and hydrogen peroxide, respectively, inhibited this luminescence. Lipopolysaccharide (LPS) did not elicit luminescence of hemocytes in relation to controls. The catalase-inhibittable phenol red oxidation assay also showed an increase in the level of hydrogen peroxide produced by hemocytes stimulated with PMA or Micrococcus luteus. LPS did not stimulate the hemocytes, similarly to the observed by luminescence assay's. We also evaluated ROS production by fluorescence microscopy and we found approximately 25% more fluorescent hemocytes when zymosam was used. This fluorescence was inhibited by catalase. In DHR fluorimetry assay we observed an increase in the intensity of fluorescence in PMA stimulated hemocytes. This fluorescence was inhibited by cystein, a reducing agent that removes hydrogen peroxide and peroxinitrite. We conclude that hemocytes of the tick, like other invertebrate such as mollusks, crustaceans and insects and vertebrate, produce reactive oxygen species during the immune response. This is the first report of reactive oxygen species production by arachnid hemocytes.
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Bruno, Trezia Ieda Ballerini. "Infecção de Biomphalaria glabrata com Angiostrongylus costaricensis : desenvolvimento larval e resposta hemocitaria". [s.n.], 2005. http://repositorio.unicamp.br/jspui/handle/REPOSIP/314681.
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Orientador: Eliana Maria Zanotti-MagalhãesTese (doutorado) - Universidade Estadual de Campinas, Instituto de Biologia
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Resumo: Experimentalmente, Biomphalaria glabrata pode ser utilizada como hospedeiro intermediário do nematódeo Angiostrongylus costaricensis, responsável pela angiostrongilíase abdominal. Esta zoonose, descrita no Brasil principalmente nos estados sulinos, pode acometer acidentalmente o homem, sendo que a infecção ocorre através da ingestão de moluscos parasitados ou água e alimentos contaminados com larvas de 3° estágio, eliminadas no muco dos moluscos hospedeiros. O objetivo deste trabalho foi estudar o desenvolvimento dos estágios larvais e o comportamento dos hemócitos na hemolinfa de B. glabrata infectada.Um total de 168 moluscos foi infectado com 120 larvas LI de A. costaricensis extraídas das fezes de camundongos Swiss albinos previamente infectados via oral sob tubagem esofágica com 6 larvas L3. Larvas de A. costaricensis foram recuperadas de 45 moluscos B. glabrata após 15, 22 e 29 dias de exposição ao parasita, através do método de Baermann, utilizando tecidos digeridos dos moluscos com solução de pepsina e ácido clorídrico. Constatou-se maior recuperação de larvas de A. costaricensis dos moluscos aos 29 dias de infecção. Para o estudo do desenvolvimento de Ao costaricensis, 60 moluscos infectados foram destinados a recuperação larval durante 30 dias consecutivos. Foi observada a mudança larval de LI para L2 aos 13 dias de infecção e L2 para L3 aos 18 dias de infecção. Hemolinfa de 45 moluscos infectados e não infectados com A. costaricensis foi coletada para verificação da resposta hemocitária durante 4 semanas. Os hemócitos foram distinguidos em hialinócitos e granulócitos. Enquanto nos moluscos não infectados predominaram os hialinócitos, naqueles infectados os granulócitos foram mais evidentes, principalmente entre o 18° ao 25° dia de infecção. Foi confirmada a ocorrência tanto da infecção percutânea como por via oral. Os locais mais parasitados foram: região cefalopodal, a preferida pelo nematódeo, seguida do intestino, rim e pulmão. Todas as larvas encontradas estavam viáveis e rodeadas por reação do tipo granulomatosa, independentes de sua localização
Abstract: Biomphalaria glabrata can be experimentally used as an intermediate host of the nematode Angiostrongylus costaricensis, responsible for abdominal angiostrongyliasis. This zoonosis, found in Brazil mainly in the southem states, can accidentally infect man through the ingestion of parasitized mollusks or contaminated water and food containing third-stage larvae, eliminated in the mucous secretion of the mollusks. The objective of this work was to study the development of larval stages and the behavior of hemocytes in the hemolymph of infected B. glabrata. A total of 168 mollusks were infected with 120 LI larvae of A. costaricensis, extracted ftom excrement of albino Swiss mice previously infected via the oral route by esophageal tube with 6 L3 larvae. The A. costaricensis larvae had been recovered from 45 B. glabrata mollusks at 15, 22 and 29 days after exposure to the parasite, by means of the method of Baermann, using molluscan tissues digested with pepsin and hydrochloric acid solution. A larger recovery of A. costaricensis larvae from the mollusk was found at 29 days after infection. For the study of the development of A. costaricensis, 60 infected mollusks were allocated for larval recovery during a period of 30 consecutive days. It was observed that there was a larval stage change, from L1 to L2, at the 13th day after infection and from L2 to L3 on the 18th day after infection.The hemolymph of 45 mollusks, both infected and not infected with A. costaricensis, was collected for verification of the hemocyte response during 4 weeks. The hemocytes were differentiated into hyalinocytes and granulocytes. While in the non infected mollusks the hyalinocytes had predominated, in those infected granulocytes were more evident, mainly between the 18th and the 25th day after infection. The occurence of infection, both via percutaneous and via oral routes, was confirmed. The most parasitized sites were the cephalopodan mass, preferred by the nematodes, folIowed by the intestines, kidneys and lungs. AlI the larvae found were viable and surrounded by reaction of the granulomatous type, independent of their situation
Doutorado
Mestre em Parasitologia
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GIAMBERINI, LAURE Pihan Jean-Claude. "ETUDE DES MECANISMES DE TRANSPORT ET DE DETOXICATION DES METAUX LOURDS CHEZ LA MOULE D'EAU DOUCE DREISSENA POLYMORPHA. ROLE DES HEMOCYTES ET DES ORGANES DU SYSTEME EXCRETEUR (HISTOLOGIE, ULTRASTRUCTURE, MICROANALYSE) /". [S.l.] : [s.n.], 1993. ftp://ftp.scd.univ-metz.fr/pub/Theses/1993/Giamberini.Laure.SMZ9321_1.pdf.
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CustÃdio, Richeyla Kelly de Assis. "Perfil ClÃnico e Laboratorial dos Pacientes com Leucemia Linfoide CrÃnica Atendidos no ServiÃo de Hematologia e Hemoterapiado HUWC-HEMOCE". Universidade Federal do CearÃ, 2009. http://www.teses.ufc.br/tde_busca/arquivo.php?codArquivo=3804.
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FundaÃÃo de Amparo à Pesquisa do Estado do CearÃA leucemia linfocÃtica crÃnica (LLC) à a leucemia mais freqÃente nos paÃses ocidentais, ocorrendo em cerca de 30% de todas as leucemias do adulto, acometendo principalmente a populaÃÃo acima de 65 anos de idade. à caracterizada pela proliferaÃÃo clonal de linfÃcitos tipo B CD5 positivos. Do ponto de vista epidemiolÃgico, raramente à vista entre os orientais e a sua incidÃncia nÃo sofre influÃncia de agentes ambientais como substÃncias tÃxicas ou radiaÃÃes ionizantes. Os pacientes com LLC podem apresentar evoluÃÃo clÃnica variÃvel desde indolente a forma mais agressiva e rapidamente fatal. Na Ãltima dÃcada, houve um grande avanÃo no conhecimento da fisiopatologia desta doenÃa e fatores de prognÃstico de maior impacto foram estabelecidos e, do ponto de vista terapÃutico, novos medicamentos com maior eficÃcia surgiram. Desta forma à interessante conhecer a real situaÃÃo desta doenÃa com caracterÃsticas epidemiolÃgicas, clÃnicas e de prognÃstico muito peculiares. Nesse trabalho traÃamos o perfil (demogrÃfico, clÃnico e laboratorial) dos pacientes com LLC atendidos no serviÃo de Hematologia e Hemoterapia do Cearà (HEMOCE) do HUWC, correlacionando a expressÃo dos marcadores de prognÃstico (CD38 e ZAP-70) com algumas variÃveis e com o tratamento farmacolÃgico. Foram utilizadas amostras de sangue de 55 pacientes com LLC confirmados atravÃs de imunofenotipagem, em tratamento ou nÃo. Os marcadores CD38 e ZAP-70 foram analisados por citometria de fluxo e a dosagem de β2-microglobulina obtida atravÃs de mÃtodo imunoenzimÃtico (ELISA). Os dados obtidos foram analisados atravÃs de programa estatÃstico. Observamos um leve predomÃnio de pacientes do sexo masculino (1,4:1), 78% acima de 65 anos de idade e 60% eram procedentes da capital. Quanto ao estadiamento, 50,9% pacientes foram classificados como de baixo risco, segundo classificaÃÃo de Rai. Quanto a evoluÃÃo clÃnica, mais de 30% dos pacientes nÃo apresentavam sinais clÃnicos, nem no momento do diagnÃstico nem no final do estudo. Em relaÃÃo aos marcadores, 22,2% dos pacientes apresentavam expressÃo do ZAP-70 e 24,13% do CD38. NÃo houve diferenÃa estatÃsticamente significante entre os marcadores ZAP-70 e CD38 e as variÃveis hematolÃgicas (Hb, Ht e Plaquetas). Houve correlaÃÃo entre o estadiamento clÃnico de Rai e o tratamento farmacolÃgico e com os marcadores de prognÃstico (ZAP-70 e CD38). O perfil foi caracterizado por pacientes em estÃgios clÃnicos, segundo Rai, de baixo risco, com ausÃncia da expressÃo dos marcadores de prognÃstico (ZAP-70 e CD38) e com ausÃncia de tratamento especÃfico.
Chronic Lymphocytic Leukemia (CLL) is the most frequent leukemia on western countries, responding for 30% off all leukemia cases, reaching, specially, the under 65 years old population. It is characterized by accumulation of neoplasic mature B lymphocytes positives CD5. Concerning epidemiology, it is barely seen among eastern people, and its incidence is not influenced by environmental agents, such as toxic substances or ionizing radiation. CLL-B patients may present a variable clinical evolution, which goes from indolent to aggressive and rapidly fast cases. On last decade, there have been great advance on biology knowledge of this disease, and greater impact prognostic factors have been established. In relation to treatment, new medication, with better efficacy, appeared. Therefore, it is interesting to know the real situation of this disease, which has singular epidemiological, clinical and prognostic characteristics. In this work we trace the demographical, clinical and laboratorial profile of patients with CLL-B, at a state hemocenter (HEMOCE). We correlate prognostic markers expression (CD38 e ZAP-70) with some variable and with pharmacological treatment. We used 55 blood samples from patients CLL-B confirmed by their immunophenotypes; these patients may or may not be in treatment. CD38 and ZAP-70 were analyzed by flow cytometry and β2-microglobulin dosage, obtained through ELISA. Results were treated by statistic program. We see a discrete prevalence of male patients (1,4:1), 78% are under 65 years old and 60% live at CearÃâs capital. In relation to Rai staging system, 50,9% were classified as low risk patients. Concerning clinical evolution, more than 30% of the patients did not show clinical signs, neither at diagnose moment nor at the end of this study. In relation to the markers, 22,2% of the patients show ZAP-70 expression, and 24,13% show CD38 expression. There was no significant statistic difference between ZAP-70 and CD38 markers and hematological variable (Hb, Ht and platelets). There was correlation between Raiâs staging and pharmacological treatment and with prognostic markers. CONCLUSION: The profile was characterized by low risk patients, according Raiâs staging, it lacks prognostic markers expression and it also lacks specific treatment.
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Custódio, Richeyla Kelly de Assis. "Perfil clínico e laboratorial dos pacientes com leucemia linfoide crônica atendidos no serviço de hematologia e hemoterapiado HUWC-HEMOCE". reponame:Repositório Institucional da UFC, 2009. http://www.repositorio.ufc.br/handle/riufc/4266.
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CUSTÓDIO, Richeyla Kelly de Assis. Perfil clínico e laboratorial dos pacientes com leucemia linfóide crônica atendidos no serviço de hematologia e hemoterapia do HUWC-HEMOCE. 2009. 79 f. Dissertação (Mestrado em Ciências Farmacêuticas) - Universidade Federal do Ceará. Faculdade de Farmácia, Odontologia e Enfermagem, Fortaleza, 2009.Submitted by denise santos (denise.santos@ufc.br) on 2013-01-04T12:27:57Z No. of bitstreams: 1 2009_dis_rkacustódio.pdf: 1909686 bytes, checksum: b1765670a26a4948113ea73dde11465a (MD5)
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Chronic Lymphocytic Leukemia (CLL) is the most frequent leukemia on western countries, responding for 30% off all leukemia cases, reaching, specially, the under 65 years old population. It is characterized by accumulation of neoplasic mature B lymphocytes positives CD5. Concerning epidemiology, it is barely seen among eastern people, and its incidence is not influenced by environmental agents, such as toxic substances or ionizing radiation. CLL-B patients may present a variable clinical evolution, which goes from indolent to aggressive and rapidly fast cases. On last decade, there have been great advance on biology knowledge of this disease, and greater impact prognostic factors have been established. In relation to treatment, new medication, with better efficacy, appeared. Therefore, it is interesting to know the real situation of this disease, which has singular epidemiological, clinical and prognostic characteristics. In this work we trace the demographical, clinical and laboratorial profile of patients with CLL-B, at a state hemocenter (HEMOCE). We correlate prognostic markers expression (CD38 e ZAP-70) with some variable and with pharmacological treatment. We used 55 blood samples from patients CLL-B confirmed by their immunophenotypes; these patients may or may not be in treatment. CD38 and ZAP-70 were analyzed by flow cytometry and β2-microglobulin dosage, obtained through ELISA. Results were treated by statistic program. We see a discrete prevalence of male patients (1,4:1), 78% are under 65 years old and 60% live at Ceará’s capital. In relation to Rai staging system, 50,9% were classified as low risk patients. Concerning clinical evolution, more than 30% of the patients did not show clinical signs, neither at diagnose moment nor at the end of this study. In relation to the markers, 22,2% of the patients show ZAP-70 expression, and 24,13% show CD38 expression. There was no significant statistic difference between ZAP-70 and CD38 markers and hematological variable (Hb, Ht and platelets). There was correlation between Rai’s staging and pharmacological treatment and with prognostic markers. CONCLUSION: The profile was characterized by low risk patients, according Rai’s staging, it lacks prognostic markers expression and it also lacks specific treatment.
A leucemia linfocítica crônica (LLC) é a leucemia mais freqüente nos países ocidentais, ocorrendo em cerca de 30% de todas as leucemias do adulto, acometendo principalmente a população acima de 65 anos de idade. É caracterizada pela proliferação clonal de linfócitos tipo B CD5 positivos. Do ponto de vista epidemiológico, raramente é vista entre os orientais e a sua incidência não sofre influência de agentes ambientais como substâncias tóxicas ou radiações ionizantes. Os pacientes com LLC podem apresentar evolução clínica variável desde indolente a forma mais agressiva e rapidamente fatal. Na última década, houve um grande avanço no conhecimento da fisiopatologia desta doença e fatores de prognóstico de maior impacto foram estabelecidos e, do ponto de vista terapêutico, novos medicamentos com maior eficácia surgiram. Desta forma é interessante conhecer a real situação desta doença com características epidemiológicas, clínicas e de prognóstico muito peculiares. Nesse trabalho traçamos o perfil (demográfico, clínico e laboratorial) dos pacientes com LLC atendidos no serviço de Hematologia e Hemoterapia do Ceará (HEMOCE) do HUWC, correlacionando a expressão dos marcadores de prognóstico (CD38 e ZAP-70) com algumas variáveis e com o tratamento farmacológico. Foram utilizadas amostras de sangue de 55 pacientes com LLC confirmados através de imunofenotipagem, em tratamento ou não. Os marcadores CD38 e ZAP-70 foram analisados por citometria de fluxo e a dosagem de β2-microglobulina obtida através de método imunoenzimático (ELISA). Os dados obtidos foram analisados através de programa estatístico. Observamos um leve predomínio de pacientes do sexo masculino (1,4:1), 78% acima de 65 anos de idade e 60% eram procedentes da capital. Quanto ao estadiamento, 50,9% pacientes foram classificados como de baixo risco, segundo classificação de Rai. Quanto a evolução clínica, mais de 30% dos pacientes não apresentavam sinais clínicos, nem no momento do diagnóstico nem no final do estudo. Em relação aos marcadores, 22,2% dos pacientes apresentavam expressão do ZAP-70 e 24,13% do CD38. Não houve diferença estatísticamente significante entre os marcadores ZAP-70 e CD38 e as variáveis hematológicas (Hb, Ht e Plaquetas). Houve correlação entre o estadiamento clínico de Rai e o tratamento farmacológico e com os marcadores de prognóstico (ZAP-70 e CD38). O perfil foi caracterizado por pacientes em estágios clínicos, segundo Rai, de baixo risco, com ausência da expressão dos marcadores de prognóstico (ZAP-70 e CD38) e com ausência de tratamento específico.
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Eliautout, Remi. "Etude de la réponse immunitaire de la cicadelle Circulifer haematoceps au cours de l'infection par Spiroplasma citri". Thesis, Bordeaux, 2014. http://www.theses.fr/2014BORD0260/document.
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Spiroplasma citri est une bactérie phytopathogène transmise par la cicadelle Circuliferhaematoceps. L'absence de symptômes malgré la multiplication de S. citri dans l'hémolymphe, suggèreque le système immunitaire joue un rôle important dans la tolérance de la cicadelle vis-à-vis duspiroplasme.Le but de cette thèse a donc été d'étudier la réponse immunitaire de C. haematoceps aucours de l'infection par S. citri.Notre étude sur le système immunitaire de la cicadelle a montré la présence dans le plasma d'uneactivité antibactérienne et d'une activité phénoloxidase. Parmi les principaux types d'hémocytes unephagocytose des bactéries par les granulocytes et les plasmatocytes a été observée. Les gènessusceptibles d'être impliqués dans ces processus ont été recherchés par une approche par hybridationsoustractive. De manière étonnante, aucun gènes codant des récepteurs ni d'effecteurs connus del'immunité n'ont été identifiés. En revanche certains gènes (23 en tout) codent des protéines ayantpotentiellement un rôle immunitaire. Six de ces 23 gènes ont été retenus pour suivre leur expressionau temps précoce d'une infection bactérienne. Les résultats ont montré que les gènes codantI'Hexamérine, la DDBPl et la Thiorédoxine peroxydase étaient surexprimés lors de l'infection par 5.citri. Une approche fonctionnelle d'interférence par ARN a montré d'une part que I'Hexamérine étaitimpliquée dans l'activité phénoloxidase et d'autre part qu'elle jouait un rôle important dans la surviede C. haematococeps au cours de l'infection par 5. citri. En parallèle, le suivi de l'activité phénoloxidaseet de la phagocytose au cours de l'infection a montré que 5. citri était capable de s'adapter à laréponse immunitaire de l'insecte et d'y échapper. Ces résultats rejoignent ceux obtenus chez ladrosophile concernant S. poulsoniiSpirop/asma citri is phytopathogenic bacteria transmitted by the leafhopper Circuliferhaematoceps. The absence of symptoms despite the multiplication of S. citri in the hemolymph,suggests that the immune system plays an important role in the tolerance of the leafhopper towardsthe spiroplasma infection. The purpose of this thesis was to study the immune response of C.haematoceps during the infection by 5. citri.The characterization of the immune system of the leafhopper showed that an antibacterial activity anda phenoloxidase activity were present in the plasma. The main types of hemocytes were identified.Among them, granulocytes and plasmatocytes are capable to phagocyte bacteria. The genes involvedin these immune processes were searched using subtractive hybridization method. lnterestingly, noneof the genes known to encode receptors or effectors of the immune system were identified. On theother hand 23 putative immune genes were identified. Six of these genes were retained to follow theirexpression in the early time of a bacterial infection. The results showed that the genes encodingHexamerin, DDBPl and Thioredoxin peroxidase were up-regulated during the infection by 5. citri. Afunctional approach by gene silencing showed that Hexamerin was involved in the phenoloxidaseactivity and played an important role in the survival of C. haematoceps during the infection by S. citri.Finally, the follow-up of the phenoloxidase activity and phagocytosis by hemocytes showed anadaptation and an evasion of S. citri from the immune response of the insect, according to the resultsobtained for 5. pou/sonii-infected drosophila.Keywords : 5piroplasma citri, phenoloxidase, phagocytosis, hemocytes, gene silencing, Hexamerine,subtractive hybridization
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Jiravanichpaisal, Pikul. "White Spot Syndrome Virus Interaction with a Freshwater Crayfish". Doctoral thesis, Uppsala : Acta Universitatis Upsaliensis : Univ.-bibl. [distributör], 2005. http://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-5776.
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Alhabshi, Manaf. "Validation of a haemoglobin measuring method for determination of blood loss at oral and maxillofacial surgical treatment". Thesis, Malmö högskola, Odontologiska fakulteten (OD), 2016. http://urn.kb.se/resolve?urn=urn:nbn:se:mau:diva-19941.
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ABSTRACTBackground: Determination of blood loss can be a crucial factor at surgical procedures, especially when the amount of blood is small and mixed with other fluids. The existing methods to measure this are still not supported with evidence enough.Aim: To validate the accuracy of the HemoCue® system (HemoCue, Ängelholm, Sweden) at estimation of blood loss in a setup where blood is mixed with saline and saliva.Materials and methods: The haemoglobin concentration of defibrinated horse blood was measured using the haematology analysers Hemocue® 201+ and Hemocue Plasma/Low Hb Photometer. Series of non-diluted blood (control) and blood diluted with saline and saliva (test) were conducted to mimic a clinical situation at oral and maxillofacial surgical treatment. Following each dilution, a new measurement of the haemoglobin concentration was performed using the appropriate haematology analyser to measure the blood loss.Results: There were no statistically significant differences regarding the measured Hb concentration in relation to the degree of dilution. The accuracy of measured blood volume in the diluted and non-diluted blood (control) was within a level of ± 11,5%.Conclusion: The measurement of haemoglobin concentrations in a mixture of blood, saline, and saliva, proved to be accurate when compared to non-diluted blood. It is suggested that the HemoCue® system can be applied in the field of oral and maxillofacial surgery to improve the accuracy of the blood loss measurement.
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Machado, RosÃngela Pinheiro GonÃalves. "Perfil clÃnico dos pacientes com LLC-B do AmbulatÃrio do Hospital UniversitÃrio Walter CantÃdio/HEMOCE: corelaÃÃo com marcadores biolÃgicos de prognÃstico". Universidade Federal do CearÃ, 2009. http://www.teses.ufc.br/tde_busca/arquivo.php?codArquivo=4594.
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IntroduÃÃo: A Leucemia LinfocÃtica CrÃnica (LLC) à uma neoplasia caracterizada pela proliferaÃÃo clonal de linfÃcitos de aspecto maduro. Apresenta curso clÃnico e prognÃstico heterogÃneo. Rai e Binet criaram sistemas de prognÃsticos clÃnicos que classificam a LLC em baixo, intermediÃrio e alto risco. Surgiram os marcadores biolÃgicos de prognÃstico que aumentaram o poder preditivo na LLC. Objetivo:Caracterizar os marcadores clÃnicos e biolÃgicos de pognÃstico dos pacientes com LLC do ambulatÃrio do Hospital UniversitÃrio Walter CantÃdio (HUWC)/Centro de Hematologia e Hemoterapia do CearÃ/HEMOCE. Metodologia: Trata-se de um estudo retrospectivo, transversal e observacional com 43 pacientes portadores de LLC, recrutados de forma randomisada, no perÃodo de agosto de 2007 a Junho de 2009. Foram coletados dos pacientes dados nos prontuÃrios, entrevista e trÃs amostras com 5,0 ml de sangue venoso perifÃrico em Ãcido etilenodiaminotetraacÃtico (EDTA) para o hemograma, por metodologia automatizada no equipamento CellDynÂ, modelo 3.500; dosagem da Ã2-microglobulina (Ã2-M) sÃrica pelo teste quantitativo automatizado no aparelho MINI- VIDAS (BioMÃrieuxÂ) e imunofenotipagem em citÃmetro de fluxo Beckman Coulter EPICS XL-MCL (Coulter). Em seguida, foi coletado pela punÃÃo da medula Ãssea o aspirado para o mielograma e 4 a 5 ml, em 2 ml de heparina, para a avaliaÃÃo citogenÃtica por bandaâG. A anÃlise dos dados foi realizada utilizando os programas estatÃsticos Biostat 4.0 e GraphPad Prism (versÃo 5.0), o teste Coeficiente de Phi e o teste Coeficiente de Contingencia C. Os testes de Fisher e Qui-quadrado com Ãndice de significÃncia α = 5%. Kaplan-Meier para funÃÃo de sobrevivÃncia e o teste log rank. Os resultados foram gerados usando o software livre R, versÃo 2.7. Resultados: Os pacientes (74,42%) tinham idade acima de 60 anos; 58,14% homens e 41,86% mulheres; a maioria (32,56%) trabalhava na agricultura; pardos (74,42%), procedentes da capital (53,49%); histÃria familiar de LLC desconhecida (46,51%); sintomÃticos ao diagnÃstico (53,49%); com comorbidades (hipertenÃÃo arterial e Diabetes Melitus) (51,16%); estÃgio 0 (34,89%), I e II (51,16%), III e IV (13,95%) de Rai; A (44,19%), B (44,19%) e C (11,62%) de Binet ; tempo de duplicaÃÃo linfocitÃra (TDL) ausente (81,40%); biÃpsia da medula Ãssea com padrÃo nÃo difuso (57,14%); a desidrogenase lÃctica (LDH) normal (83,72%), avaliados ao diagnÃstico. Os exames obtidos durante a evoluÃÃo dos pacientes revelaram um perfil imunofenotÃpico clÃssico de LLC- B, com expressÃo de CD5+, CD19+, CD23+ e imunoglobulina de superfÃcie de baixa expressÃo; a maioria com Zap-70 negativa (77,50%); expressÃo de CD38 negativa (73,81%); Ã2-M aumentada (55,81%); cariÃtipo normal (44,4%) e alteraÃÃes genÃticas em 11, 11% pela citogenÃtica clÃssica. As curvas de sobrevidas dos pacientes com Zap-70 e CD38 negativos apresentaram maior tempo de sobrevida livre de tratamento. ConclusÃo:Os pacientes avaliados eram idosos, com tendÃndia ao diagnostico tardio decorrente do contexto socioeconÃmico; apresentaram LLC indolente, pelos critÃrios de estadiamentos clÃssicos (Rai, Binet, TDL, padrÃo da histologia da medula Ãssea, LDH) e biolÃgicos (as expressÃes da Zap-70 e CD38), exceÃÃo à Ã2-M, porÃm, sem significÃncia. Aqueles com Zap-70 e CD38 negativos apresentaram maior sobrevida livre de tratamento. Pacientes do sexo masculino apresentaram evoluÃÃo e prognÃstico semelhantes ao feminino. O tratamento prevalente foi clorambucil associado à prednisona e nÃo levou os pacientes à remissÃo clÃnica ou hematolÃgica. Os marcadores de prognÃsticos tenderam à correlaÃÃo na identificaÃÃo dos pacientes dentro dos subgrupos de riscos.Introduction: Chronic lymphocytic leukemia (CLL) is a neoplasm characterized by clonal proliferation of lymphocytes of mature appearance. Clinically and prognostic heterogeneous. Rai and Binet established clinical prognostic systems that classify LLC in low, intermediate and high risk. Soon, the biological markers of prognosis that increased the predictive power of the LLC. Objective: To characterize the clinical and biological markers of pognÃstico of patients with CLL the outpatient department of a university hospital (HUWC) / Center for Hematology and Hemotherapy Cearà / HEMOCE). Methodology: This is a retrospective, cross-sectional and observational 43 patients LLC, recruited so randomisation, from August 2007 to June 2009. We collected patient data from medical records, interview and three samples with 5.0 ml of peripheral venous blood in ethylenediaminetetraacetic acid (EDTA) for blood, for automated methodology CellDyn  equipment, model 3500, measurement of Ã2-microglobulin (Ã2 - M) serum by automated quantitative test on the device MINI-VIDAS (BioMÃrieux Â) and immunophenotyping on flow cytometry Beckman Coulter  EPICS XL-MCL (Coulter). Then collect the puncture of bone marrow aspirate and bone marrow examination for 4 to 5 ml in 2 ml of heparin for cytogenetic evaluation by Banda - G. Data analysis was performed using the statistical programs Biostat 4.0 and GraphPad Prism (version 5.00), the Phi coefficient test and the test coefficient Contingency C. The Fisher and chi-square test with significance level α = 5%. Kaplan-Meier survival function and log rank test. The results were generated using the free software R, version 2.7. Results: The patients (74.42%) were aged over 60 years, 58.14% 41.86% men and women, the majority (32.56%) worked in agriculture; brown (74.42%), coming the capital (53.49%), family history of unknown LLC (46.51%), symptomatic at diagnosis (53.49%), with comorbidity (arterial hypertension and Diabetes Mellitus) (51.16%), stage 0 ( 34.89%), I and II (51.16%), III and IV (13.95%) Rai, A (44.19%), B (44.19%) and C (11.62%) of Binet, lymphocyte doubling time (SRT) absent (81.40%), bone marrow biopsy with non-diffuse pattern (57.14%), lactate dehydrogenase (LDH) normal (83.72%), valued at diagnosis. The tests obtained during the course of the patients showed an immunophenotypic profile of classic B-CLL with expression of CD5 +, CD19 +, CD23 + surface immunoglobulin and low-expression, most with Zap-70 negative (77.50%); expression CD38 negative (73.81%), beta-2 microglobulin increased (55.81%), normal karyotype (44.4%) and genetic alterations in 11, 11% by classical cytogenetics. Survival curves of patients with Zap-70 negative and CD38 showed longer survival free of treatment. Conclusion: The patients studied were elderly, to encourage improve with late diagnosis due to the socioeconomic context, LLC indolent presented by classical staging criteria (Rai, Binet, TDL, standard bone marrow histology, LDH) and biological (the expression of Zap -70 and CD38), except for beta-2 microglobulin, but without statistical significance. Those with Zap-70 and CD38 negative had higher survival free of treatment. Male patients showed progress and prognosis similar to female. The prevalent treatment was associated with chlorambucil prednisone and did not lead patients to clinical remission or hematologic. The prognostic markers of the correlation tended to identify patients within the subgroups of risk.
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Wu, Pin-Yi, e 吳品逸. "Separation of Penaeus monodon Hemocyte--Development and Application of Monoclonal Antibodies against Hemocyte Subpopulation". Thesis, 1996. http://ndltd.ncl.edu.tw/handle/51355751861814774011.
Texto completo da fonteResumo:
碩士東吳大學
微生物學系
85
In shrimp, hemocytes constitute mainly the first line of defence against the pathgenic infections. In order to realize the defence functions of hemocytes, they must be separated into the different subpopulations based on the cell surface markers. in order to get the monoclonal antibodies (Mabs)against the granular cells (GC), hemocyte from tiger shrimp (Penaeus monodon) were separated by a negative selection with concanalin A (Con A)-linked bead. It could rise up the Gc content from 31% to 78% in the total hemocytes. FourMabs, which were disignated as Z5E10 (IgG2a), Z6A5 (IgG1), Z6A6 (IgG1), andZ6H8 (IgG2b), were produced via the Balb/c mouse immunized with the hemocytes with a rich GC content. Their reactivities to hemocytes were assayed with the enzymeimmuno staining. The molecular mass of the corresponding epitopes weredetermined with either the Western-blotting or the immunoprecipitation. A 14kDa protein, present in both GC and semigranular cells (SGC), could be recognized by Mab Z5E10, no matter was in a reducing or a non- reducing forms. The molecular mass of the non-reducing epitopes, present in SGC, were largerthan 205 kDa which was determined by the recognition of the other there Mabs. Epitope recognized by Mab Z6A6 was show to be a glycoprotein proven with themethod of periodate oxidation, however, those epitopes recognized by the other three Mabs were protein in nature. These results imply that four Mabs could recognize different epitopes of the hemocytes. Besides, observing the indirect immunofluorescence with the confocal microscope, antigens thatdistributed on the hemocytes could cross-link to the capping or ring forms once they were recognized by the four Mab. Following the negative selection with each of the four Mab-Link beads, the concentration of hyaline cells increased in hemocyte suspension. These results suggested that antigens recognized by the four Mabs were surface location.