Tesis sobre el tema "Laser capture microdissection"
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Stuart, Charles A., William L. Stone, Mary E. A. Howell, Marianne F. Brannon, H. Kenton Hall, Andrew L. Gibson y Michael H. Stone. "Myosin Content of Individual Human Muscle Fibers Isolated by Laser Capture Microdissection". Digital Commons @ East Tennessee State University, 2015. https://dc.etsu.edu/etsu-works/4642.
Texto completoMajithia, Haritika. "Determining cell-specific gene expression in two soybean mutants using laser capture microdissection". Thesis, McGill University, 2013. http://digitool.Library.McGill.CA:80/R/?func=dbin-jump-full&object_id=119666.
Texto completoSoya, Glycine max, (L.) Merr., est généralement couvert de trichomes et possède trois folioles par feuille composée. Deux cultivars de soya mutant, un sans trichomes, cv. Glabrous, et un avec cinq folioles par feuille composée, cv. 5-LF, ont été comparés avec un cultivar sauvage pour étudier la différence dans l'expression des gènes. Comme les trichomes se développent et se différencient depuis l'épiderme et comme le sort des feuilles (qu'elle devienne composée ou simple) se décide au niveau du méristème, l'expression des gènes des cellules spécifiques de l'épidermes a été comparée au méristème dans les trois cultivars via un instrument de microdissection au laser ainsi qu'à l'aide de séquençage d'ARN à haute capacité. Les résultats indiquent qu'environ 200 gènes distincts dans les deux tissues (méristème et épiderme) ont été exprimés différemment dans chacun des trois cultivars. Le méristème avait une expression plus élevée de domaines de liaison à l'ADN spécifiques de séquence alors que l'épiderme avait une plus forte expression de gènes liés à la défense des plantes.
Signour, Thomas. "Extraction de signatures de bactéries par microspectroscopie Raman et chimiométrie : application à l’étude de la composition biologique des aérosols dans l’environnement". Thesis, Lille 1, 2017. http://www.theses.fr/2017LIL10152/document.
Texto completoFor several years, the study and the control of the quality of the air are at the heart of all the concerns. In 2012, the DGA (Direction Générale de l’Armement) employs the ASTRID program (Accompagnement Spécifique des Travaux de Recherches et d’Innovation Défense), to accompany the dual civil and military research work. This thesis is part of this approach and proposes the feasibility study, by Raman microspectroscopy, of the concept of rapid detection and identification of microorganisms present in an air sample, with a resolution at the species level. For this, we construct a chemometric model for the classification of micro-organisms representative of the natural biodiversity. Such a model is built by acquiring, without a priori i) the Raman spectra of these microorganisms after biocollection; and ii) the genomic sequences encoding the 16S RNAs of these same microorganisms. The research presented in this thesis therefore presents the different studies carried out during the development of a new protocol allowing the analysis of bacteria from natural environmental aerosols. We demonstrate the need to optimize the acquisition of Raman spectra on bacteria and the statistical processing of spectral data that allows the development of classification models with high recognition rates
Beasley, Brooke, Aubrey Sciara, Tiffani Carrasco, Gregory Dr Ordway y Michelle Dr Chandley. "Laser Capture Microdissection Analysis of Inflammatory-Related Alterations in Postmortem Brain Tissue of Autism Spectrum Disorder". Digital Commons @ East Tennessee State University, 2019. https://dc.etsu.edu/asrf/2019/schedule/34.
Texto completoTan, Jing [Verfasser] y Thomas [Akademischer Betreuer] Klopstock. "Laser capture microdissection of single muscle fibers for mitochondrial proteomic investigations / Jing Tan ; Betreuer: Thomas Klopstock". München : Universitätsbibliothek der Ludwig-Maximilians-Universität, 2019. http://d-nb.info/1205664874/34.
Texto completoOrdway, Gregory A., Attila Szebeni, Michelle M. Duffourc y Katalin Szebeni. "Laser Capture Microdissection and RT- PCR Analyses of Specific Cell Types in Locus Coeruleus From Postmortem Human Brain". Digital Commons @ East Tennessee State University, 2007. https://dc.etsu.edu/etsu-works/8624.
Texto completoOrdway, Gregory A., Attila Szebeni, Craig A. Stockmeier, Michelle M. Duffourc y Katalin Szebeni. "Glial Deficits in the Noradrenergic Locus Coeruleus in Major Depression Revealed by Laser Capture Microdissection and Quantitative PCR". Digital Commons @ East Tennessee State University, 2008. https://dc.etsu.edu/etsu-works/8625.
Texto completoBartel, Jan [Verfasser]. "Laser Capture Microdissection in Paraffin eingebetteter Gewebe als Werkzeug zur Bestimmung des Sialylierungsstatus von ausgewählten Zellpopulationen / Jan Bartel". Gießen : Universitätsbibliothek, 2017. http://d-nb.info/1141574675/34.
Texto completoJumper, Natalie. "Application of a site-specific in situ approach to keloid disease research". Thesis, University of Manchester, 2016. https://www.research.manchester.ac.uk/portal/en/theses/application-of-a-sitespecific-in-situ-approach-to-keloid-disease-research(f0a9bcae-93f0-4335-8839-afa5747f40d6).html.
Texto completoOrdway, Gregory A., Attila Szebeni, Michelle M. Duffourc, Sophie Dessus-Babus y Katalin Szebeni. "Gene Expression Analyses of Neurons, Astrocytes, and Oligodendrocytes Isolated by Laser Capture Microdissection From Human Brain: Detrimental Effects of Laboratory Humidity". Digital Commons @ East Tennessee State University, 2009. https://dc.etsu.edu/etsu-works/8606.
Texto completoMoncada, Benavides Camilo Andres. "EFFECT OF NICOTINE ON LUNG S-ADENOSYLMETHIONINE AND PNEUMOCYSTIS PNEUMONIA DEVELOPMENT". Diss., Temple University Libraries, 2012. http://cdm16002.contentdm.oclc.org/cdm/ref/collection/p245801coll10/id/206623.
Texto completoPh.D.
Infection with "Pneumocystis" causes a ≥ 99% depletion of plasma S-adenosylmethionine (AdoMet) levels in both "Pneumocystis" pneumonia (PcP) animal models and patients. AdoMet is a critical cellular metabolic intermediate, with a pivotal role as methyl donor in a myriad of biochemical processes and necessary for the synthesis of the essential polyamines spermidine and spermine. In the target tissue of "Pneumocystis", the lung, levels of AdoMet were previously shown to be depleted experimentally using nicotine. Here we show that chronic administration of nicotine in an animal model of PcP resulted in decreased lung AdoMet content. Since "Pneumocystis" is dependent on this metabolite, PcP burden was also relived. We hypothesized that the underlying mechanism behind nicotine-induced AdoMet depletion was an increased consumption of AdoMet through the polyamine pathway where the increased activity of N-1-spermidine/spermine acetyl transferase raises the catabolic / anabolic cycling of polyamines, a process that utilizes AdoMet. In a critical test of our hypothesis, we found that blockage of polyamine metabolism via inhibition of the polyamine biosynthetic enzyme ornithine decarboxylase (ODC) hinders the effect of nicotine on lung AdoMet levels. Further support is provided by metabolite analyses showing nicotine to cause a strong diversion of AdoMet toward polyamine synthesis and away from methylation reactions; these shifts are also reversed by inhibition of ODC. Because the nicotine effect on "Pneumocystis" is so striking, we considered the possibility of tissue specificity. Using laser capture microdissection (LCM), we collected samples of lung alveolar regions (site of infection) and respiratory epithelium for controls. We found nicotine to cause increased ODC activity in alveolar regions but not airway epithelium; we conclude that tissue specificity likely contributes to the effect of nicotine on "Pneumocystis" pneumonia. Our studies demonstrate the feasibility of pharmacological manipulation of the polyamine pathway in order to reduce AdoMet levels in the lung and prompted the assessment of compounds alternative to nicotine with the potential to achieve a comparable effect. In vitro evaluation of the polyamine analog DENSPM along with putrescine in type II alveolar cell lines, indicates that although such a combination has the potential to induce polyamine flux, an apparent competition for the same polyamine transport system impairs simultaneous uptake of both compounds at effective concentrations. In conclusion, we showed that chronic nicotine administration causes reduction of AdoMet levels in rat lung following 21 days of treatment, by a mechanism involving the induction of polyamine flux, which is responsible of increased AdoMet utilization for polyamine biosynthesis. According to LCM-based analysis, this effect seems to be confined to the alveolar regions of the lung.
Temple University--Theses
Thwe, Le Myo. "Biomarker Analysis in Paediatric Tumours Diagnosed within A Single Institution". Thesis, The University of Sydney, 2016. http://hdl.handle.net/2123/16297.
Texto completoOrdway, Gregory A., Attila Szebeni, Michelle J. Chandley, Craig A. Stockmeier, Michelle M. Duffourc y Katalin Szebeni. "Abnormal Gene Expression in Noradrenergic Neurons and Surrounding Glia in Brains of Depressed Suicide Victims Revealed by Laser Capture Microdissection and qPCR". Digital Commons @ East Tennessee State University, 2009. https://dc.etsu.edu/etsu-works/8628.
Texto completoMolina, Mariana. "Abordagem para análise proteômica de neurônios contendo neuromelanina na substância negra, isolados por microdissecção a laser". Universidade de São Paulo, 2015. http://www.teses.usp.br/teses/disponiveis/5/5160/tde-22012016-104704/.
Texto completoCurrently the worldwide proportion of people aged 60 years and over is growing faster than any other age group. This strikingly epidemiological transition results in an increase of aging related diseases, including Parkinson\'s disease (PD). Although many brain areas exhibit the neuropathological signs of Parkinson\'s disease, the degeneration of neuromelanin containing cells in the substantia nigra is considered a hallmark feature, representing cardinal diagnostic criteria for PD. However, why certain brain regions -- such as the substantia nigra -- are vulnerable in some neurodegenerative diseases, while some neighboring morphologically indistinguishable neurons remain preserved, is still unclear. Molecular analysis of specific neuronal populations can lead us to a better understanding about the physiological role played by these neurons and mechanisms involved in disease\'s processes. In a post-genomic era, proteomic analyses are of great scientific interest since they allow a better understanding of the biological processes. The laser capture microdissection technique has also became an important tool in biological research, being increasingly used for acquisition of pure populations of cells from histological sections, preventing the dissection of areas outside the target tissue. The combination of these methods can significantly contribute to understand the pathophysiological role of the containing neuromelanin neurons of the substantia nigra. However, for an effective application of both techniques, laser capture microdissection and proteomic analysis, it is essential the application of an efficient protocol. In this context, this study aims to establish a protocol for laser microdissection of containing neuromelanin neurons in cognitively normal individuals for subsequent proteomic analyses. We selected cases from the Brain Bank of the Brazilian Aging Brain Study. A collaboration with the Medical Proteome Center, University of Bochum, Germany took part during the development of our proposal. Our protocol was developed based on previous published protocols and optimized according the intended aims. We analyzed anatomical planes for neuronal collection, freezing methods, thickness of tissue for microdissection sections, solution for tissue collection during laser microdissection and the proteolytic digestion methods. Through our comparative tests, we have achieved the desired results and validated them by mass spectrometry analyses. Consequently, we were also able to exclude technical factors that could possibly preclude one effective proteome analysis
Fischer, Anthony John. "Augmenting antiviral host defense in the respiratory epithelium". Diss., University of Iowa, 2009. https://ir.uiowa.edu/etd/500.
Texto completoOrdway, Gregory A. "Abnormal Gene Expression in Noradrenergic Neurons and Surrounding Glia in Brains of Depressed Suicide Victims Revealed by Laser Capture Microdissection and qPCR". Digital Commons @ East Tennessee State University, 2009. https://dc.etsu.edu/etsu-works/8666.
Texto completoOwens, Misty. "BDNF-Related Gene Expression of Laser Capture Microdissected Glutamate Neurons from the Anterior Cingulate Cortex in Mouse Models of Autism Spectrum Disorder". Digital Commons @ East Tennessee State University, 2020. https://dc.etsu.edu/etd/3805.
Texto completoOwens, Misty. "BDNF-Related Gene Expression of Laser Capture Microdissected Glutamate Neurons from the Anterior Cingulate Cortex in Mouse Models of Autism Spectrum Disorder". Digital Commons @ East Tennessee State University, 2008. https://dc.etsu.edu/etd/3805.
Texto completoCrawford, Jessica D. "Cellular-based Brain Pathology in the Anterior Cingulate Cortex of Males with Autism Spectrum Disorder". Digital Commons @ East Tennessee State University, 2014. https://dc.etsu.edu/etd/2443.
Texto completoStuart, Charles A. y Michael H. Stone. "Reply to "Letter to the Editor: Comments on Stuart Et Al. (2016): 'myosin Content of Individual Human Muscle Fibers Isolated by Laser Capture Microdissection'"". Digital Commons @ East Tennessee State University, 2016. https://dc.etsu.edu/etsu-works/4675.
Texto completoMacedo, Carolina Carneiro Soares 1989. "Análise da expressão diferencial de proteínas em ilhas neoplásicas grandes e pequenas por espectrometria de massas baseada em proteômica e sua relação com o prognóstico". [s.n.], 2015. http://repositorio.unicamp.br/jspui/handle/REPOSIP/289257.
Texto completoDissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Odontologia de Piracicaba
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Resumo: O carcinoma espinocelular oral (CEC) é o tipo de neoplasia maligna mais comum em cabeça e pescoço, com alta prevalência e morbidade. O tratamento é baseado em sistemas de classificação pouco precisos e o prognóstico é ruim em muitos casos. Diferentes padrões histológicos já foram descritos na tentativa de melhor compreender o curso da doença, predizer prognóstico e auxiliar no tratamento. As diferentes áreas do tumor apresentam características morfológicas e moleculares distintas resultando em comportamentos clínicos específicos, e estudos recentes apontam a região de invasão tumoral (do inglês invasive front tumor) como área de interesse para análises de perfil molecular e identificação de possíveis marcadores de prognóstico. O padrão de invasão neoplásico tem relação com a agressividade tumoral e a presença de ilhas no fronte invasivo já foi descrita como pior padrão. Assim, o objetivo desta dissertação foi comparar a composição diferencial de proteínas totais de ilhas neoplásicas grandes e pequenas do fronte e do interior do tumor, e correlacionar essas proteínas com o prognóstico. A proteômica foi associada à microdissecção a laser (ML), consideradas juntas como ferramentas de alta robustez para identificação de proteínas em tecidos neoplásicos em regiões específicas. Vinte peças cirúrgicas de CEC oral de língua fixadas em parafina foram submetidas a ML para obtenção das amostras compostas pelas seguintes regiões do tecido: 1) ilhas neoplásicas grandes da região frontal, 2) ilhas neoplásicas pequenas da região frontal, 3) ilhas neoplásicas grandes do interior do tumor e 4) ilhas neoplásicas pequenas do interior do tumor, seguida pela extração de proteínas e análise por cromatografia líquida acoplada à espectrometria de massas (LC-MS/MS). A anotação funcional das proteínas e a correlação aos dados clínico-patológicos dos pacientes foram realizadas. Foram identificadas 1906 proteínas totais, sendo 1480 proteínas comuns entre as quatros regiões estudadas. Duas proteínas foram exclusivas nas ilhas grandes do fronte e sete nas ilhas grandes do interior. Oitenta e cinco proteínas foram diferencialmente expressas entre a região do fronte e o interior tumoral, e destas, 57 foram relacionadas a dados clínico-patológicos importantes para o prognóstico. Os processos biológicos, como desenvolvimento da epiderme, adesão celular, apoptose, ciclo celular, degradação da matriz extracelular e expressão gênica, evidenciados entre as proteínas diferencialmente expressas confirmam as mudanças moleculares associadas à progressão neoplásica. A combinação de ML, MS e bioinformática foi capaz de identificar um painel de proteínas que podem auxiliar a desvendar o curso do CEC oral, predizendo agressividade e prognóstico. Em acréscimo, essa abordagem pode ajudar ainda na compreensão das diferenças e dos mecanismos de sinalização entre diferentes áreas do tecido
Abstract: Oral squamous cell carcinoma (SCC) is the most common type of malignant tumor in head and neck, with high prevalence and morbidity. Treatment is based on inaccurate classification systems and the prognosis is poor in many cases. Different histological patterns have been described in an attempt to better understand the course of the disease, predict prognosis and assist in treatment. Different areas of the tumor have different morphological and molecular characteristics resulting in specific clinical behaviors, and recent studies point to the region of tumor invasion as an area of interest for molecular profile analysis and identification of possible prognostic markers. The pattern of neoplastic invasion is related to tumor aggressiveness and the presence of islands in the invasive front has been described as worst invasion pattern. The objective of this work was to compare the protein differential expression of large and small islands neoplastic from the front and the inner tumor, and to correlate these proteins with prognosis. Proteomics was associated with laser microdissection (ML), and together they are considered robustness tools to identify proteins in neoplastic tissues in specific regions of interest. Twenty surgical specimens of oral tongue SCC fixed in paraffin were subjected to ML to obtain sample composed by the following regions of tissue: 1) large neoplastic frontal islands, 2) small neoplastic islands of the frontal region, 3) large neoplastic islands inside the tumor and 4) small islands within the neoplastic tumor, followed by extraction and analysis of proteins by liquid chromatography coupled to mass spectrometry (LC-MS/MS). The functional annotation of proteins and correlation with clinicopathological data from patients were performed. A total of 1906 proteins were identified, with 1480 common proteins between the four regions studied. Two proteins were exclusives in the large islands of the forehead and seven in large islands in the interior. Eighty-five proteins were differentially expressed between the front region and inner tumor, and of these, 57 were related to clinical and pathological data. The biological processes such as development of the epidermis, cell adhesion, apoptosis, cell cycle, disassembly of the extracellular matrix and gene expression, evidenced among the differentially expressed proteins confirmed the molecular changes associated with neoplastic progression. The combination of ML, MS, and bioinformatics was able to identify a panel of proteins that may help to unravel the course of oral SCC, predicting aggressiveness and prognosis. In addition, this approach may also help in understanding the differences and signaling mechanisms between different areas of the tumor tissue
Mestrado
Patologia
Mestra em Estomatopatologia
Li, Su-Chen. "Small Intestinal Neuroendocrine Tumor Analyses : Somatostatin Analog Effects and MicroRNA Profiling". Doctoral thesis, Uppsala universitet, Endokrin Onkologi, 2014. http://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-233207.
Texto completoKollbeck, Sophie L. G. [Verfasser], Jens [Akademischer Betreuer] Habermann y Frank [Gutachter] Köster. "Vergleichende Proteomanalyse zwischen Colitis Ulcerosa assoziierten und sporadischen kolorektalen Karzinomen nach Laser Capture Mikrodissektion : Comparative proteomic analysis of Ulcerative Colitis associated and Sporadic Colorectal Cancer after Laser Capture Microdissection / Sophie L. G. Kollbeck ; Gutachter: Frank Köster ; Akademischer Betreuer: Jens Habermann". Lübeck : Zentrale Hochschulbibliothek Lübeck, 2020. http://d-nb.info/1207574155/34.
Texto completoFigueiredo, Joana Maria Serra de Oliveira Duarte. "The role of microRNAs in amyotrophic lateral sclerosis". Master's thesis, Faculdade de Ciências e Tecnologia, 2011. http://hdl.handle.net/10362/7991.
Texto completoMicroRNAs (miRNAs) are emerging as a primary mediator of gene regulation in many different cell types. There is increasing evidence that specific subsets of miRNA play a prominent role in the nervous system, both in development and in specific neurodegenerative diseases. This study aims to elucidate the role of microRNA in selective motor neuron death that is the hallmark of amyotrophic Lateral sclerosis (ALS). Pre-symptomatic time-point was chosen since the levels of miRNAs are highly likely to be altered as a secondary consequence of cell injury and death in ALS. Laser capture microdissection (LCM) was used to study miRNA profiles in motor neurons of spinal cord tissue from SOD1G93A mice, the best characterized model of ALS. In preliminary work, using miRNA specific chips we have identified 2 miRNAs which are dramatically upregulated before disease onset. In this study, high RNA quality was achieved from laser captured cells, which consist in a major advance towards obtaining meaningful results of these miRNAs expression in downstream applications. Despite LCM technology has become increasingly sophisticated; rapidly obtaining enough amount of starting material for downstream applications is still extremely challenging. The combination of this optimized technique with microarrays, followed by RT-qPCR may provide insights into potential contribution of microRNAs to progression of neurodegeneration of motor neurons in ALS.
Majer, Anna. "Temporal deregulation of genes and microRNAs in neurons during prion-induced neurodegeneration". Virology Journal, 2010. http://hdl.handle.net/1993/30718.
Texto completoOctober 2015
Almeida, Marcia Rodrigues de. "Bases moleuculares da recalcitrância ao enraizamento adventício em Eucalyptus globulus Labill". reponame:Biblioteca Digital de Teses e Dissertações da UFRGS, 2015. http://hdl.handle.net/10183/131932.
Texto completoEucalyptus is one of the most planted tree species in the world today, mainly due to its use as raw material for paper, cellulose and wood industries. Eucalyptus globulus and its hybrids have low lignin contents and are of great interest to industry, as this feature facilitates and reduces costs of the cellulose extraction process. However, this species is recalcitrant to adventitious rooting, making vegetative propagation by cuttings difficult. Aiming at a better understanding of the molecular mechanisms involved in rooting recalcitrance in E. globulus, this study analyzed changes in morphological, anatomical and molecular patterns during adventitious rooting in this species. Exogenous auxin exposure reversed the recalcitrant phenotype in E. globulus, significantly increasing rooting percentage. The vascular cambium was identified as a region of auxin accumulation and also the site from where adventitious roots originated. Gene expression analysis in cambium cells indicated that TOPLESS and IAA12, auxin signaling repressors, and ARR1, involved in cytokinin signaling pathway, appear to act as negative regulators of adventitious rooting. The high expression of those genes in control plants was significantly decreased by exogenous auxin treatment. Comparatively, in an easy-to-root species, E. grandis, the expression of these genes was significantlylower in both treatment conditions, and the concentration of endogenous indole- 3-acetic acid in control plants was higher. Analysis of the protein pattern during rooting in E. globulus plants treated or not with exogenous auxin allowed the identification of proteins involved in diverse biological processes, mainly oxidative stress and energy metabolism. Interesting differences were identified when comparing different rooting conditions or phases. Several proteins were clearly associated with the respective plant phenotype in each situation, particularly considering control plants. These results represent relevant advances in the knowledge about adventitious rooting in woody plants and can be used as tools in the design of strategies aiming at improving adventitious rooting in recalcitrant genotypes of industrial value.
Consentino, Laurent. "Mécanismes d'acquisition du fer de l'hôte chez Bacillus cereus : rôle du couple bacillibactine-FeuA et expression des gènes impliqués dans l'homéostasie du fer in vivo durant l’infection intestinale chez l’insecte". Thesis, Université Paris-Saclay (ComUE), 2019. http://www.theses.fr/2019SACLA018/document.
Texto completoIron acquisition is essential for most living organisms, including many pathogenic bacteria. However, free iron is toxic: it is bound into storage or transport proteins (e.g. ferritin, hemoproteins…) and iron homeostasis is tightly regulated. To scavenge iron from these sources, bacteria possess several systems to acquire the bound iron, by surface proteins or siderophores. Bacillus cereus is a sporeforming Gram-positive bacterium, opportunistic human pathogen, 2nd cause of food-borne disease in France. It has been demonstrated that the B. cereus surface protein IlsA and the siderophore bacillibactin (BB) are involved in iron acquisition from ferritin and that these two molecules are important for infection of the insect model G. mellonella. My thesis project focused on two parts: first the study of the BB-Fe3+ complex import into the cell by the siderophore binding protein FeuA highlights the central role of both BB and FeuA. The deletion of the genes encoding for these two molecules limits iron acquisition by B. cereus from ferritin, heme, hemoglobin and inorganic iron in vitro. On the other hand, the virulence phenotype during intra-haemocelic infection of G. mellonella is similar to the Wild-type strain. These results suggest a possible feedback on the expression of virulence factor genes when B. cereus is unable to synthetize both BB and FeuA, and therefore are under high stress. The second part of my work focused on the expression of genes involved in iron homeostasis in vivo, during gut infection of germ-free larvae of G. mellonella. We chose to perform a microgenomic approach, using laser-capture microdissection to get small samples in targeted areas, and then analysing the expression of chosen genes by RT-qPCR and ddPCR at two time points post ingestion The results show that : i) the colonisation of G. mellonella gut is impacted when B. cereus is deprived of both BB and FeuA ; ii) ilsA is expressed during gut infection ; iii) iron homeostasis is involved in adaptation and pathogenicity from the early step of infection of the insect gut ; iv) only weak gene expression modulation occured between the two timepoints This work gives new fundamental knowledge about B. cereus iron homeostasis, and highlights the use of new techniques regarding the in situ study of host-pathogen interactions
Solimena, Michele, Anke M. Schulte, Lorella Marselli, Florian Ehehalt, Daniela Richter, Manuela Kleeberg, Hassan Mziaut et al. "Systems biology of the IMIDIA biobank from organ donors and pancreatectomised patients defines a novel transcriptomic signature of islets from individuals with type 2 diabetes". Springer, 2017. https://tud.qucosa.de/id/qucosa%3A33350.
Texto completoDreja, Tanja S. "Microarray-basierte Expressionsanalysen des weißen Fettgewebes der NZO-Maus sowie der Langerhansschen Inseln der NZL-Maus : zwei Modelle für das metabolische Syndrom". Phd thesis, Universität Potsdam, 2009. http://opus.kobv.de/ubp/volltexte/2009/3237/.
Texto completoOverweight and obesity cause insulin resistance and increase the risk of developing type 2 diabetes and cardiovascular diseases. Both, obesity and susceptibility to diabetes, are to a major part genetically predisposed. The relevant genes, their interaction with the environment – especially with food components – and the pathomechanisms causing insulin resistance and diabetes are not fully known yet. In the present study the development and progression of obesity and type 2 diabetes should be investigated by the means of gene expression analyses of the white adipose tissue (WAT) and the islets of Langerhans to identify underlying pathomechanisms and new causative candidate genes. For this purpose diet intervention studies on NZO- and related NZL-mice – two polygenic mouse models for the human metabolic syndrome – were performed. A carbohydrate containing high fat-diet (HF: 14.6 % fat) caused early obesity, insulin resistance and type 2 diabetes in both mouse models. A fat reduced standard chow (SD: 3.3 % fat) which strongly delayed the onset of obesity and diabetes, and a diabetes protective carbohydrate free high fat-diet (CHF: 30.2 % fat) served as control diets. Using microarray technology genome wide expression profiles of the WAT were generated. Pancreatic islets were isolated by the means of laser capture microdissection (LCM) and expression profiles of them were created, too. Differentially expressed genes were validated by quantitative real time PCR. The HF-diet reduced the expression of nuclear genes of the oxidative phosphorylation and lipogenic enzymes in the WAT of the NZO-mouse. This suggests an inadequate storage and utilization of fat in these animals. This is specific for the obese NZO-model and wasn’t observed for the lean SJL-mouse, indicating a role in the development of insulin resistance. Additionally, there was proof that the enlargement of the WAT triggers a retarded infiltration of macrophages into the WAT and there a local immune response. Moreover, the LCM technique was established and used for the isolation of highly enriched RNA from islets of Langerhans. For the first time the influence of carbohydrates in a high fat-diet on the expression profile of pancreatic islets was investigated by the use of genome wide expression analyses at an early time point at the onset of diabetes. Contrary to the WAT the diabetogenic HF-diet in islets cells increased the expression of both nuclear genes coding for the oxidative phosphorylation and genes associated with cell proliferation. Furthermore 37 already annotated genes correlated with diabetes progression were identified. The peptide hormone cholecystokinin (Cck: 11.8-fold enriched by the HF-diet) is one of the most up-regulated genes. The strong enrichment of Cck-mRNA in islets suggests a previously unknown function of the hormone in the regulation of the islet cell proliferation. The transcription factor ChREBP (Mlxipl: 3.8-fold reduced by the HF-diet) is one of the most down-regulated genes in the islets of Langerhans. Moreover, ChREBP, which has been already identified as a glucose regulated transcription factor for lipogenic enzymes in the liver but not in islets of Langerhans, was detected for the first time in islet cells, using immunohistochemistry. This points to an until now unknown regulatory function of ChREBP in the glucosesensor mechanism of the islet cells. Correlation of the differentially expressed genes associated with diabetes progression with gene variants from human genome wide association studies for type 2 diabetes (WTCCC, Broad-DGI-T2D-study) made the identification of 24 new diabetes candidate genes possible. The results of the genome wide expression analyses, which were done for the first time on a polygenic mouse-model, corroborated previous results for monogenic mouse-models for obesity and diabetes (e.g. ob/ob- and db/db-mice), however also demonstrated differences in some instances. Especially the results concerning the oxidative phosphorylation could be relevant for the comprehension of the pathogenesis of the polygenic human metabolic syndrome.
Maniero, Carmela. "A functional study on novel genes involved in regulating aldosterone secretion in normal human zona glomerulosa and in aldosterone-producing adenomas". Thesis, University of Cambridge, 2017. https://www.repository.cam.ac.uk/handle/1810/270313.
Texto completoScheel, Tobias. "Die B-Zell-Antwort im Synovialgewebe von Patienten mit Rheumatoider Arthritis". Doctoral thesis, Humboldt-Universität zu Berlin, Mathematisch-Naturwissenschaftliche Fakultät I, 2009. http://dx.doi.org/10.18452/16026.
Texto completoAlthough B cells have an important impact on the pathogenesis of Rheumatoid arthritis (RA) still surprisingly little is known about their activation and differentiation within the inflamed synovial tissue (ST). A hallmark of RA is the presence of auto-antibodies (auto-Ab). However, still little is known about the frequency of self reactive synovial lymphocytes and it is unclear to which extent the inflamed ST contributes to auto-Ab production. These thesis deals with the characterization of the synovial B cell response and the specificity of synovial B lymphocytes. B and plasma cells (PC) from RA patients were isolated either by Laser Capture Microdissection or by FACS and their immunoglobulin(Ig)-repertoire was determined. The analysis of the VH-genes revealed that both naïve and memory B cells can immigrate the ST. A comparison of VDJ-rearrangements of B cells and PC showed that in ST without ectopic germinal centres mainly memory B cells become activated, expand clonally and differentiate into PC. During this process B cells can switch their Ig-class but do only hypermutate slightly. To determine the specificity of synovial B lymphocytes, recombinant Ab from synovial B cells and PC were generated. The polyreactivity assay showed that particularly naïve B cells were polyreactive. In contrast, the frequency of autoreactive memory B cells and PC was much higher than that of naïve B cells. In addition, Ab specific for bacterial antigens (especially for periodontal bacterias) and for the autoantigen MCV were identified. The affinity measurement of the MCV-specific autoantibody revealed that auto-Ab secreted in the ST can exhibit very high affinities. The data presented here show that B cells seem to play an important role in the maintenance and possibly the development of RA.
Oliveira, Gabriela Pintar de. "Análise da participação das células neuronais e não-neuronais na Esclerose Lateral Amiotrófica em camundongos transgênicos para SOD1 humana utilizando técnicas de microdissecção a laser e PCR em tempo real". Universidade de São Paulo, 2014. http://www.teses.usp.br/teses/disponiveis/5/5138/tde-06062014-101803/.
Texto completoAmyotrophic Lateral Sclerosis (ALS) is an adult onset motor neuron neurodegenerative disease that leads to the progressive loss of muscular functions. It is a fast progression disorder (2 to 5 years) culminating in death by respiratory failure. Recent findings suggest that non neuronal cell types, especially astrocytes and microglia, might contribute to the neuronal death. The transgenic mouse SOD1G93A, carring human mutant SOD1, was used in this study. Behavioral studies pointed to the onset of the clinical symptoms occurring at 90 days in the animal model, thus, allowing the selection of the pre-symptomatic ages of 40 and 80 days to the molecular studies. Gene expression analysis of transgenic mice and their non-transgenic littermates at those ages was performed by using a microarray platform containing the whole mouse genome and has detected 492 and 1105 differentially expressed genes at 40 days and 80 days old mice, respectively. These results were validated by quantitative PCR (qPCR). Bioinformatic analysis of the results identified 17 and 11 over-represented molecular pathways at 40 days and 80 days, respectively. Of these, endocytosis, glutamatergic synapse, ubiquitin-mediated proteolysis, chemokine signaling pathway, oxidative phosphorylation, antigen processing and presentation and also tight junction were common to both ages. Furthermore, glutamatergic synapse and fagosome were suggested as potentially more important at 40 and 80 days, respectively. Specific transcripts were analyzed on enriched samples of cells (astrocytes, microglia and motor neuron) obtained by laser microdissection from the ventral horn of mouse spinal cord. The transcripts Cxcr4, Slc1a2 and Ube2i were evaluated by qPCR in enriched samples of astrocytes of the 40 days old mice, and Cxcr4 and Slc17a6 were analyzed in motor neuron samples at this age. Cxcr4 has been found decreased in astrocytes from transgenic mice and increased in the motor neurons of these animals. Slc1a2, Ube2i and Slc17a6 have increased in the cell type in which they were evaluated in the transgenic mice. Tap2 and Tuba1a were evaluated at microglia enriched samples of 80 days old mice and were found to be increased. Finally, Akt1 has decreased in enriched samples of motor neurons from 80 days old mice. The results suggest that glutamatergic signaling might play essential role in early stages of the disease (40 days), while in phases closer to the appearance of the symptoms (80 days), the neuroimmunomodulation takes place. Thus, this study points to new perspectives for ALS study
Rasiah, Krishan Kumar St Vincent's UNSW. "The identification of novel biomarkers in the development and progression of early prostate cancer". Awarded by:University of New South Wales. St Vincent's, 2006. http://handle.unsw.edu.au/1959.4/24187.
Texto completoOeschger, Franziska M. "Subplate populations in normal and pathological cortical development". Thesis, University of Oxford, 2011. http://ora.ox.ac.uk/objects/uuid:686d99bd-36e0-47f2-9680-9874f413d1bb.
Texto completoIyer, Chitra C. "The Role of Muscle and Nerve in Spinal Muscular Atrophy". The Ohio State University, 2016. http://rave.ohiolink.edu/etdc/view?acc_num=osu1451568269.
Texto completoChen, Wen-Chi y 陳文奇. "Bacterium Isolation Using Laser Capture Microdissection". Thesis, 2008. http://ndltd.ncl.edu.tw/handle/yn2bvr.
Texto completo國立臺北科技大學
光電工程系研究所
96
Bacterial cultures are very important study issues in microbiology. Human can’t observe bacterial cultures by naked eyes, thus, the way to study bacterial cultures is totally unique. In traditional, to study unknown bacterial cultures must purify the target bacterium from complex sample conditions. In nature, bacterial cultures are mixed and coexisted. There are among of 99% of the sea bacterial cultures can’t be isolated by culture. In this study, we use laser capture microdissection to isolated a sea bacterial culture, named acinetobacter venetianus which already had been well purified by culture, and to ensure that LCM system can be used in bacterial cultures isolation. we set up LCM system on a headstand microscope, and isolate bacterial cultures under microscope view. Before isolation, we dye bacterial samples easier to be observed. We find out bacterium has been Gram’s dyed can’t be observed on translated film after isolaed in microscope view. Thus, we add DAPI on Gram’s dyed sample and redo experience again. In this study, we discover that bacterial cultures about 1μm in size can be isolated and observed under the condition that the distance between laser probe and film less than 1.0μm and laser light must irradiate translate film more than 400ms
Tseng, Yu-Wei y 曾昱瑋. "The Study of Multi-Layer Transfer Film for Laser Capture Microdissection". Thesis, 2008. http://ndltd.ncl.edu.tw/handle/q6yw4a.
Texto completo國立臺北科技大學
光電工程系研究所
96
The purpose of the study is to provide the development of multi-layer transfer film for laser capture microdissection. Before developing single material of the transfer film thickness of 30 μm, it has been successfully applied to the LCM system. In the study, we hope could reduce the film’s thickness for LCM system, and uses support layer, multi-layer optical film and uses EVA solution of two melt index values to develop new hot-melt film for the usage of LCM system. By using EVA, polyester film, TiO2 and SiO2 as hot-melt film, the support layers, and major multi-layer optical materials. By way of mixing different MI value EVA solution, substrate preparation, and spin coating etc., we made various hot-melt film with different thickness. In this study, hot-melt film’s appraisal which used AFM, SEM, and LCM system, and found the best surface roughness, hot-melt depth, and different thickness and size of the melting point to provide LCM system in different sections of the target application.
Lin, Chia-Ching y 林佳慶. "Construction of a Electric Moving Stage for Laser Capture Microdissection System". Thesis, 2008. http://ndltd.ncl.edu.tw/handle/gj984x.
Texto completo國立臺北科技大學
光電工程系研究所
96
We construct a new Laser Capture Microdissection (LCM) system by Using Electric Moving Stage. It spends a lot of time in positioning when the new LCM system with near-field fiber probe is integrated with microscopy. For overcoming the shortcoming, we designed a precision stage for moving the sample to adjust the target to be in the center of the scope. The fiber probe was set over the view center, and allowed to move only up and down. The designed stage includes a stepping motor and piezo actuators for moving fine in X-Y directions. We build a component of Human Machine Interface on electric moving stage control system and LCM system by Visual Basic. Then, we actually use electric moving stage on LCM experiments. By statistical analysis, we obtained the average displacement value of the stage is 3.265 μm and Relative standard deviation(RSD) is 0.00025. We successfully melted the EVA transfer film into figure by using the 70 nm fiber probe. The target can be moved to the center of scope in one minute by this new stage, then the process of laser capture be completed more quickly and higher precision control.
Al-Sowaimel, Lujain Fawzi. "Regional podocyte isolation using laser capture microdissection for molecular profiling of glomerular disease". Thesis, 2014. https://hdl.handle.net/2144/15219.
Texto completoShen, Chuan-Yu y 沈傳宇. "The Study of Transfer Film for Micron and Nano Laser Capture Microdissection System". Thesis, 2006. http://ndltd.ncl.edu.tw/handle/5a47m5.
Texto completo國立臺北科技大學
光電工程系所
94
The purpose of the study is to provide the innovation of transfer film for Micron and Nano Laser Capture Microdissection (LCM) system. The principle of LCM comes from placing a transfer film on the top of dissected tissue. As laser beam pierce on the transfer film, the penetrated spot then converts and absorbs laser energy into heat to adhere the target cell after melting. Then the target cell will be identified after transfer film is removed from the dissected tissue. In order to make it flat and convenient to use, transfer film is coated on the bottom of a cap. But the cap may cause negative impacts such as power loss after the absorption and reflection of laser beam. Therefore, this study is aiming to find the new type EVA cap for the usage of traditional and probe type Laser Capture Microdissection. The melting point of the transfer film is 88℃, which may damage cells easily. We use local-made EVA, which melting point is 65℃, as major material and add near-infrared absorbing dyes to enhance the laser energy absorption. By combining different concentration of EVA solution, substrate preparation, and spin-coating etc., we made various transfer films with different concentration and thickness. In this study, we designed a hollow cap, which made the fiber probe exert close melting, and the optimal conditions of making the thermoplastic film adhere on the cap were also studied.
Yen, Chi-Fu y 顏畿府. "To Construct a Laser Capture Microdissection System in Nano-Scale by Using Near-Field Fiber Probe". Thesis, 2005. http://ndltd.ncl.edu.tw/handle/3jeqp5.
Texto completo國立臺北科技大學
光電工程系所
93
Laser Capture Microdissection(LCM) is a technique that permits rapid and reliable procurement of pure population of cells from tissue sections. In LCM, the resolution of capture is directly related to the spot size on Ethyl Vinyl Acetate(EVA) transfer film. Microscope objective is used to focus laser beam and the resolution is 5μm in conventional system. In this paper, we try to update the resolution of capture by replacing objective with a near-field fiber probe. By using our new construct of LCM system, we successfully melted the EVA transfer film into spots, whose diameter ranged from 400nm to 1.4μm and the maximum droopy depth is 282.5nm. We also found the droopy depth will increase 9.53nm as laser pulsing time increases each 20ms. In the condition of laser pulsing time is 100ms and the distance between fiber probe and EVA transfer film is 1μm, we successfully captured monolayer of gold particles into spots and the minimal spot size is 400nm. These results prove that we have completed a new LCM system with higher resolution in this paper.
Xu, Baogang Jonathan. "Combining laser capture microdissection and MALDI mass spectrometry for tissue protein profiling methodology development and clinical applications /". Diss., 2005. http://etd.library.vanderbilt.edu/ETD-db/available/etd-03092005-132210/.
Texto completoChang, Chao-Yuah y 張昭元. "Selective isolation of hepatocytes by laser capture microdissection for molecular analysis in a rat model of sepsis". Thesis, 2004. http://ndltd.ncl.edu.tw/handle/69726388215659043218.
Texto completo高雄醫學大學
醫學研究所碩士班
92
It is known that proinflammatory cytokines: TNF-α, IL-1, IL-6 and adhesion molecules: P-selectin and E-selectin play roles in multiple organ dysfunction in sepsis. Carbon monoxide ( CO ) participates in regulation of cardiovascular physiology. CO is synthesized by heme-oxygenase ( HO ): inducible HO-1 or constitutive HO-2. To better understand gene expression correlated with patho-physiological functions of liver in sepsis, rats were subjected to cecal ligation and puncture ( CLP ) to induce sepsis. Rats were randomized into 4 groups of 6 animals each: CLP 9h ( early sepsis ), CLP 18h ( late sepsis ), sham 9h and sham 18h. Rat livers were excised and processed for frozen sections. To compare with conventional homogenization from liver slice, the techniques of laser capture microdissection ( LCM ) were used to isolate cells from liver sections. RNA was extracted from both liver slice and isolated specific cells by LCM. Gene expression of TNFα, IL-1β, IL-6, P-selectin, E-selectin, HO-1 and HO-2 was detected by using two-step techniques of reverse transcriptase-polymerase chain reaction ( RT-PCR ) . In situ hybridization ( ISH ) was also employed to examine gene expression in liver. In groups of LCM combined with RT-PCR, the results showed that HO-1 gene expression in early sepsis appeared significantly higher than that in both late sepsis and control ( sham 9h ). HO-1 gene expression in late sepsis appeared the same as control ( sham 18h ). IL-6 gene expression in early and late sepsis appeared significantly higher than that in sham control. In groups of tissue homogenization combined with RT-PCR, the results showed that IL-6 gene expression in early and late sepsis appeared significantly higher than that in sham control. No obvious change of HO-1, TNFα and HO-2 mRNA expression was detected among 4 groups of rats. Unlike the expression of the HO-1, IL-6, TNFα and HO-2, mRNA expression of P-selectin, E-selectin and IL-1was not detectable in any of the samples from either LCM or tissue homogenization. The results of ISH showed that gene expression of IL-6 appeared in both early and late sepsis but not in sham control. The results of ISH also showed that gene expression of HO-1 appeared in not only Kupper cells but hepatocytes in early sepsis. These studies indicate that detection sensitivity of LCM combined with RT-PCR is higher than tissue homogenization combined with RT-PCR. These studies also indicate that when rats were subjected to CLP to induce sepsis, IL-6 gene expression appeared significantly higher in early and late sepsis than sham control. HO-1 gene expression elevated in early sepsis and declined in late sepsis.
Williams, Rachel, Irene Castellano-Pelicena, Aaiad H. A. Al-Rikabi, Stephen K. Sikkink, Richard Baker, Kirsten Riches-Suman y M. Julie Thornton. "Laser capture microdissection on surgical tissues to identify aberrant gene expression in impaired wound healing in type 2 diabetes". 2020. http://hdl.handle.net/10454/18481.
Texto completoThe global prevalence Type 2 diabetes mellitus (T2DM) is escalating at a rapid rate. Patients with T2DM suffer from a multitude of complications and one of these is impaired wound healing. This can lead to the development of non-healing sores or foot ulcers and ultimately to amputation. In healthy individuals, wound healing follows a controlled and overlapping sequence of events encompassing inflammation, proliferation, and remodelling. In T2DM, one or more of these steps becomes dysfunctional. Current models to study impaired wound healing in T2DM include in vitro scratch wound assays, skin equivalents, or animal models to examine molecular mechanisms underpinning wound healing and/or potential therapeutic options. However, these do not fully recapitulate the complex wound healing process in T2DM patients, and ex vivo human skin tests are problematic due to the ethics of taking punch biopsies from patients where it is known they will heal poorly. Here, a technique is described whereby expression profiles of the specific cells involved in the (dys)functional wound healing response in T2DM patients can be examined using surplus tissue discarded following amputation or elective cosmetic surgery. In this protocol samples of donated skin are collected, wounded, cultured ex vivo in the air liquid interface, fixed at different time points and sectioned. Specific cell types involved in wound healing (e.g., epidermal keratinocytes, dermal fibroblasts (papillary and reticular), the vasculature) are isolated using laser capture microdissection and differences in gene expression analyzed by sequencing or microarray, with genes of interest further validated by qPCR. This protocol can be used to identify inherent differences in gene expression between both poorly healing and intact skin, in patients with or without diabetes, using tissue ordinarily discarded following surgery. It will yield greater understanding of the molecular mechanisms contributing to T2DM chronic wounds and lower limb loss.
European Commission 7th Framework Programme for Research and Technical Development - Marie Curie Innovative Training Networks (ITN), Grant agreement no 607886. Aveda, Hair Innovation & Technology, USA
McGrice, Hayley Ann. "Molecular characterisation of primary wool follicle initiation in Merino sheep". 2010. http://hdl.handle.net/2440/60986.
Texto completohttp://proxy.library.adelaide.edu.au/login?url= http://library.adelaide.edu.au/cgi-bin/Pwebrecon.cgi?BBID=1523639
Thesis (Ph.D.) -- University of Adelaide, School of Agriculture, Food and Wine, 2010
Braig, Ute Christine [Verfasser]. "Untersuchung genetischer Aberrationen am duktal invasiven Mammakarzinom mittels laser capture microdissection (LCM) und comparativer genomischer Hybridisierung : Etablierung einer neuen Methode / vorgelegt von Ute Christine Braig". 2003. http://d-nb.info/966347552/34.
Texto completoCoelho, Mafalda Maria Pereira. "Avaliação do conhecimento atual dos profissionais de saúde oral quanto ao diagnóstico do cancro oral". Master's thesis, 2016. http://hdl.handle.net/10284/5390.
Texto completoIntroduction: Currently oral cancer is the sixth most common cancer worldwide. Its high rate of morbidity and consequent mortality, is related with the detection of this disease already in advanced stages of malignancy. In this context, it ends up leading to increased complications in treatment that becomes more aggressive according to the degree of malignancy. Therefore early diagnosis along with suitable treatment is the fundamental key to control the disease. Due to this reality, health professionals have a key role in early detection of potential malignant or malignant lesions making it a priority goal in health. To evaluate this kind of lesions, the definitive histopathologic diagnosis provided by conventional biopsy, still remains the crucial method of choice for health professionals, although new non-invasive techniques have emerged as auxiliary diagnosis methods. Objective: This paper aims to obtain a detailed knowledge of oral cancer, including its main particularities and specificities in order to obtain an early detection and consequently a favorable prognosis. It has also have been conducted a study that collected information on the general knowledge of portuguese dentists and stomatologists regarding oral cancer, the use of auxiliary diagnostic tools nowadays available, as well as data collection concerning the clinical performance protocol when suspected potentially malignant or malignant lesions. Materials and Methods: This study was based on the keywords and outlined specific inclusion and exclusion criteria, in order to raise and discuss as much information about this great association between oncology and dentistry, which is oral cancer. In order to meet the stated goals, it was associated with this monography a quantitative study of a descriptive and cross-sectional type. Data collection was conducted through a survey involving 324 dental professionals. The survey, with a total of twenty-one questions, aimed to collect information on the general knowledge of dentists regarding oral cancer. After data collection, these were processed and analyzed using computer programs such as Microsoft Office Excel (2010) and the Statistical Package for Social Sciences (SPSS IBM © © Statistics) vs. 22.0 for Windows. Results: The selected sample has demonstrated the necessary and basic knowledge when approached about the subject, consciously being alert if suspected a potentially malignant or malignant lesion. About 78% of respondents prefer to refer the patient when faced with a suspected lesion. Regarding the individuals that perform biopsy, 66.7% prefer the excisional technique. 96% approaches other colleagues when suspects of a malignant lesion. Most professionals (88.9%) reported ever used other auxiliary methods in addition to biopsy. Of the 11.1% who admit to having used an auxiliary diagnostic tool, 58.8% indicates the toluidine blue as the preferred method. 27.2% of professionals do not consider the biopsy as a fundamental element. Discussion and Conclusion: Through the research made, it was found that it is essential to have knowledge of signs and symptoms and provide screening in routine consults aiming for an early detection of potentially malignant or malignant lesions. By general consensus of those surveyed, it was found that the clinical history, clinical examination and biopsy are the most effective methods and commonly used to reduce morbidity and mortality associated with this disease. Still, 27.2% of the respondents do not consider the biopsy as a fundamental element, which is concerning. However it was corroborated that the majority of respondents although theoretically able, prefer to refer instead of performing biopsy.