O típico sabor de chocolate é único, obtido somente de sementes fermentadas, secas e torradas de cacau, não podendo ser sintetizado artificialmente. O desenvolvimento desse sabor é influenciado pela constituição genética das sementes, processamento pós-colheita e manufatura. Proteínas dos cotilédones são potencialmente precursores do sabor e aroma de chocolate. O presente trabalho teve como objetivo analisar diferenças qualitativas e quantitativas nas proteínas de sementes de três genótipos de Theobroma cacao após a colheita e durante a fermentação, de forma a correlacionar estes resultados com diferenças na qualidade (sabor e aroma) obtidas por análise sensorial. Um dos desafios foi o isolamento de proteínas das sementes, evitando o alto teor de polifenóis e polissacarídeos que interferem na separação das proteínas e na análise do proteoma. A metodologia de extração composta por filtração em Miracloth, solubilização e precipitação em ácido ticloroacético (TCA) apresentou géis de maior resolução e repetibilidade, tendo sido escolhida como metodologia de extração protéica para estudo das alterações no proteoma das sementes de cacau durante a fermentação. Foi necessária também a utilização de kit comercial de purificação de proteínas e utilização de método de coloração com nitrato de prata para garantir géis com resolução dos spots e repetibilidade satisfatórias. Os spots foram isolados e após digestão tríptica, submetidos ao sequenciamento por cromatografia líquida associado ao espectrômetro de massas. Os espectros foram analisados pelo programa MASCOT MS/MS Ion Search, utilizando bancos de dados do NCBI. Análises dos mapas 2-D mostraram variação no número de spots entre as variedades. Ao final da fermentação, as proteínas ainda presentes nas sementes das variedades SIAL 70 e Catongo eram ácidas, e o processo de degradação foi caracterizado pelo desaparecimento de quase todas as proteínas neutras ou básicas e também de algumas proteínas ácidas; as proteínas com massa molar acima de 35 kDa também foram todas degradadas. Na variedade CCN 51, não ocorreu o mesmo perfil degradativo, havendo proteínas até pI 6,5 e massa molar acima de 100 kDa. Dos cem spots submetidos ao sequenciamento, 89 foram identificados. As proteínas 21kDa e vicilina foram as proteínas mais abundantes nos cotilédones. Correlacionando os resultados da análise sensorial e a proteômica concluiu-se que existe correlação tanto quantitativa como qualitativa das proteínas dos cotilédones de cacau e possivelmente com as proteínas precursoras de sabor de chocolate / Typical chocolate flavor is unique, only obtained from fermented, dried and roasted cocoa seeds, and can not be synthesized artificially. The flavor development is influenced by the genetic constitution, post-harvest processing and manufactures. Cotyledons proteins are believed to be the precursors of the chocolate flavor. The aim of the present work was to analyze qualitative and quantitative protein differences in seeds of three cocoa genotypes after harvesting and during the fermentation and to correlate these results with differences in quality (flavor and aroma) obtained by sensorial evaluation. One of the challenges was the isolation of proteins from cocoa seeds, avoiding the high content of polyphenols and polysaccharides which disturb protein separation and proteome analysis. The methodology of extraction by filtration in Miracloth, solubilization and precipitation in trichloroacetic acid showed the highest gel resolution and reprodutivity, and, thus, was chosen to be used in the analyses of the proteome of cocoa seeds during the fermentation. It was also necessary to use commercial kit for protein purification and a silver-based staining method with nitrate to guarantee gels with spots resolution and satisfactory reproducibility. Proteins were excised from de gels and after tryptic digestion, MS analysis was conducted by on line chromatografhy using a Cap-LC coupled to a mass spectrometer. The spectra were processed using MASCOT MS/MS Ion Search, and the sequences searched against NCBI databases. The 2-DE maps analysis of cocoa seeds showed significant variation of the spots number among the genotypes. At the end of fermentation, proteins still present in the Sial 70 and Catongo genotypes were acid and the degradation process was characterized by the disappearance of almost all the neutral or basic proteins and also some acid proteins. The genotype CCN 51 did not show the same degradation profile. Of the spots submitted to the mass spectrometer, 89 were identified. The 21kDa protein and vicilin were the most abundant proteins in the cocoa cotyledons. Correlating sensorial analysis and the proteomic results we could observe the existence of quantitative as qualitative correlation of proteins from cocoa cotyledons and possibly with the precursors proteins of chocolate flavor

A proteína prion celular (PrPC) consiste em uma glicoproteína de membrana que atua como receptora para diversas moléculas, desencadeando sinais intracelulares. Ao interagir com a co-chaperona STIP1, PrPC promove a autorrenovação e proliferação de células-tronco/progenitoras neurais (NSPCs) durante a fase embrionária. De fato, PrPC tem se destacado por sua participação na neurogênese embrionária e adulta, porém o papel de sua interação com a proteína STIP1 na neurogênese adulta permanece obscuro. Deste modo, o presente trabalho adotou abordagens in vitro para avaliação do complexo PrPC-STIP1 em processos celulares que culminam na neurogênese adulta. Para isso, culturas primárias de NSPCs de camundongos deficientes (Prnp-/-) e tipo-selvagens (Prnp+/+) para PrPC foram realizadas, e a cultura foi devidamente padronizada e caracterizada. Através de ensaios de autorrenovação, proliferação e migração celular sugere-se que PrPC promove estes eventos celulares independentemente de STIP1, e que possivelmente a proteína laminina seja um alvo crítico para migração via PrPC. / Cellular prion protein (PrPC) consists in a membrane glycoprotein that acts as a receptor to several molecules, triggering intracellular signals. By interacting with co-chaperone STIP1, PrPC promotes self-renewal and proliferation of neural stem/progenitor cells (NSPCs) during embryonic stage. Indeed, PrPC has excelled for its participation in embryonic and adult neurogenesis, but the role of its interaction with STIP1 protein in adult neurogenesis remains unclear. Thus, herein it was adopted in vitro approaches in order to evaluate the PrPC-STIP1 complex on cellular processes that culminate in adult neurogenesis. In order to assess that, NSPC primary cultures of PrPC deficient (Prnp-/-) and wild-type (Prnp+/+) mice were performed, and the culture was properly standardized and characterized. Through self-renewal, proliferation and cell migration assays, it was suggested that PrPC promotes these cellular events regardless of STIP1, and possibly the laminin protein is a critical target for migration via PrPC.

Made available in DSpace on 2014-06-11T19:22:15Z (GMT). No. of bitstreams: 0 Previous issue date: 2008-06-18Bitstream added on 2014-06-13T18:07:56Z : No. of bitstreams: 1 barberis_lrm_me_botfca.pdf: 692407 bytes, checksum: e9fd31bfed979387901d4ea3d7ea99a0 (MD5) / Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) / O presente trabalho teve como objetivo desenvolver tecnologias para o uso de plantas como unidades de produção ou diretamente como fontes de agentes fotossensibilizantes do Ácido 5-Aminolevulinato (5-ALA), Protoporfirina IX (Proto IX) e precursores destes compostos em plantas. Os experimentos foram conduzidos no Núcleo de Pesquisas Avançadas em Matologia (NUPAM) – FCA – UNESP - Botucatu/SP, durante o ano de 2007. Foram realizados ensaios para seleção do melhor conjunto de condições para o acúmulo de protoporfirina IX e seus precursores em alface, milho e cana-de-açúcar, cujas variáveis analisadas foram: 1) seleção de comprimentos de onda; 2) duração dos períodos com e sem luz; 3) variedades (11 genótipos); 4) aplicação de antioxidantes (vitaminas C e E); 5) compostos que interferem na síntese de Proto IX (oxyfluorfen, carfentrazone e ácido levulênico); 6) adição de precursores da síntese de Proto IX (glutamato); 7) substituição da atmosfera com supressão do Oxigênio. Os resultados foram analisados sobre a dispersão das médias dos tratamentos em relação às parcelas testemunhas para os tratamentos e espécies estudadas. Utilizou-se a técnica de HPLC-MS para identificação e quantificação dos compostos. A partir dos resultados dos ensaios realizados foram selecionados três experimentos: I) Acúmulo de Proto IX e 5-ALA a partir de inibidores da protoporfirinogênio oxidase (Protox) e fontes seletivas de luz (Escuro, Sombrite 75%, Claro+Vitamina C+E, Claro, Filtro Azul, Filtro vermelho, Filtro Amarelo, Filtro Verde, Testemunha) em plantas de alface, pulverizadas com Oxyfluorfen + Glutamato monossódico + Vitamina C e E, portanto 9 tratamentos com 4 repetições. II) Seleção de genótipos de cana-de-açúcar (Saccharum officinarum) para acúmulo de Proto IX com uso de herbicidas inibidores da Protox (Oxyfluorfen e Carfentrazone)... / The present work had as objective to develop technologies for the use of plants about units of production or directly as sources of agents photosensitizers, Acid 5- Aminolevulinate (5-ALA), Protoporphyrin IX (Proto IX) and precursors this compounds in plants. The experiments were conducted at the Center for Advanced Research in Weed Plant (NUPAM) – FCA – UNESP - Botucatu/SP, during the year 2007. Tests were conducted to select the best set of conditions for the accumulation of protoporphyrin IX and precursors in lettuce, corn and sugar cane, whose variables were analyzed: 1) selection of a wavelength, 2) duration of periods with and without light, 3) varieties (11 genotypes), 4) application of antioxidants to reduce the action of Proto IX (vitamins C and E), 5) compounds that interfere with the synthesis of Proto IX (oxyfluorfen, carfentrazone and levulênic acid), 6) addition of precursors for synthesis of Proto IX (glutamate), 7) Replacement of the atmosphere with suppression of the Oxygen. The results were analyzed on the dispersion of the averages of the treatments in relation to the portions witness for the treatments and studied species. It was used technique of HPLC-MS for identification and quantification of compounds. From the results of tests conducted three experiments were selected: I) Accumulation of Proto IX and 5- ALA from protoporphyrinogen oxidase (Protox) inhibitor selective and sources of light (Dark, Shade 75%, Light + Vitamin C + E, Light, Blue filter, Red filter, Yellow filter, Green filter, Witness) in lettuce plants, sprayed with Monosodium Glutamate + Oxyfluorfen + Vitamin C and E, so 9 treatments with 4 repetitions. II) Selection of genotypes of sugarcane for accumulation of Proto IX with the use of herbicides Protox inhibitors (Oxyfluorfen and Carfentrazone), precursors of Proto IX (Glutamate and Levulenic Acid) and antioxidants (vitamin C and E) in 8 genotypes... (Complete abstract click electronic access below)

Ocean-continent convergence and subsequent continental collision are responsible for continental growth, mountain building, and severe tectonic events including volcanic eruptions and earthquake activity. They are also key driving forces behind the extensive thermal and compositional heterogeneities at crustal and mantle depths. Active subduction along the Calabrian Arc in southern Italy and the Hellenic Arc are examples of such collisional tectonics. The first part of this thesis examines the subduction related deformations within the crust beneath the southern Apennines. By modeling regional surface wave recordings of the largest temporary deployment in the southern Apennines, a lower-crustal/upper-mantle low-velocity volume extending down to 50 km beneath the mountain chain is identified. The magnitude (~ 0.4 km/s slower) and anisotropic nature (~ 10%) of the anomaly suggest the presence of hot and partially molten emplacement that may extend into the upper-crust towards Mt. Vulture, a once active volcano. Since the Apulian basement units are deformed during the compressional and consequent extensional events, our observations favor the ``thick-skin'' tectonic growth model for the region. In the deeper mantle, active processes are thermodynamically imprinted on the depth and strength of the phase transitions. This thesis examines more than 15000 SS precursors and provides the present-day reflectivity structure and topography associated with these phase transitions. Through case studies I present ample evidence for both slab penetration into the lower mantle (beneath the Hellenic Arc, Kurile Island and South America) and slab stagnation at the bottom of the Mantle Transition Zone (beneath the Tyrrhenian Sea and eastern China). Key findings include (1) thermal anomalies (~ 200 K) at the base of the MTZ, which represent the deep source for Cenozoic European Rift Zone, Mount Etna and Mount Cameroon volcanism, (2) significant depressions (by 20-40 km) at the bottom of the Mantle Transition Zone beneath subducting slabs, (3) a strong 520-km reflector near subducting slabs, (4) a weak and elevated (15-25 km) 410-km reflector within active deformation zones, (5) strong lower mantle reflectors (~ 900 km) while slabs penetrate into the lower mantle, and (6) consistency between the topography of a 300-km reflector and an exothermic phase transformation. / Geophysics

Murine neural precursor cells have been a well studied model for neural cell fate determination and stem cell function both in vivo and in primary culture. However, factors such as cell number, the presence of multiple cell populations and of niche intrinsic factors made it difficult to dissect the mechanisms regulating cortical development. To overcome this issue, we have developed a culture system where mouse embryonic stem cells (ES) are differentiated to cortical radial precursors through retinoic acid treatment of embryoid bodies. One day after plating in neural differentiation conditions, ~70% of cells in the culture are cortical radial precursors (RPs) as indicated by the definitive cortical marker Emx1, and over 8 days in culture, these RPs differentiate to pyramidal glutamatergic neurons of the cortex mimicking in vivo development. Astrocyte differentiation can be observed later as the culture progresses, which again mimics the typical timed genesis of cells in the cortex. The stem cell properties and cell fate of these RPs can be manipulated with growth factors in culture as they are in vivo. In particular, FGF2 promotes proliferation and survival, while ciliary neurotrophic factor (CNTF) induces precocious astrocyte formation. Thus, our ES-derived cortical RP cultures can serve as an alternate and complementary in vitro model to examine neural precursor biology during early development.

Murine neural precursor cells have been a well studied model for neural cell fate determination and stem cell function both in vivo and in primary culture. However, factors such as cell number, the presence of multiple cell populations and of niche intrinsic factors made it difficult to dissect the mechanisms regulating cortical development. To overcome this issue, we have developed a culture system where mouse embryonic stem cells (ES) are differentiated to cortical radial precursors through retinoic acid treatment of embryoid bodies. One day after plating in neural differentiation conditions, ~70% of cells in the culture are cortical radial precursors (RPs) as indicated by the definitive cortical marker Emx1, and over 8 days in culture, these RPs differentiate to pyramidal glutamatergic neurons of the cortex mimicking in vivo development. Astrocyte differentiation can be observed later as the culture progresses, which again mimics the typical timed genesis of cells in the cortex. The stem cell properties and cell fate of these RPs can be manipulated with growth factors in culture as they are in vivo. In particular, FGF2 promotes proliferation and survival, while ciliary neurotrophic factor (CNTF) induces precocious astrocyte formation. Thus, our ES-derived cortical RP cultures can serve as an alternate and complementary in vitro model to examine neural precursor biology during early development.

El objetivo de este trabajo ha sido el estudio de dos factores genéticos implicados en la diferenciación de las neuronas GABAérgicas de proyección estriatales: Nolz1 e Ikaros. Además se ha estudiado el potencial de estos factores de transcripción en la diferenciación de células madre neurales in vitro hacia un fenotipo estriatal para entender mejor los mecanismos que regulan la aparición de este tipo de neuronas.Hemos demostrado que Nolz1 es un gen que interviene en la especificación de los precursores neurales de la SVZ de la eminencia ganglionar lateral (LGE), los llamados progenitores basales. Las células Nolz1 positivas provienen de precursores positivos para Gsh2, y su expresión contribuye a la aparición de la señalización de ácido retinoico en el núcleo estriado durante el desarrollo. Nolz1, además, regula la vía de Notch en precursores neurales i promueve la aparición de precursores oligodendrogliales mediante un mecanismo no autónomo en la LGE. Por otra parte, demostramos que la isoforma de Ikaros Ik1 es esencial para la correcta generación de las neuronas encefalinérgicas estriatales a partir de la diferenciación de precursores neurales Dlx2 positivos de la LGE. Mostramos que Ik1 es necessario para sacar de ciclo los precursores que formarán el núcleo estriado mediante el incremento de los niveles del inhibidor de ciclinas p21. En consonancia con esta observación, la falta de Ikaros durante el desarrollo, afecta la neurogénesis tardía en el estriado. Esto se traduce en que el animal deficiente de Ikaros presenta un núcleo estriado más pequeño debido a una disminución en el número de neuronas de proyección encefalinérgicas de la matriz. Así, Nolz1 e Ikaros intervienen en diferentes momentos del desarrollo estriatal, siendo el papel de Nolz1 anterior y centrado en el control de precursores neurales, y el de Ikaros algo posterior, centrado en la diferenciación de neuronas estriatales de proyección encefalinégicas / The main goal of this work has been the study of two genetic factors implied in the differentiation of the striatal GABAergic projection neurons: Nolz1 and Ikaros. Moreover, we have studied the potential of these transcription factors to direct the differentiation of neural stem cells in vitro towards a striatal phenotype, in order to understand some aspects of the generation of this kind of neurons.We have demonstrated that the action of Nolz1 is involved in the specification of the neural precursors which reside in the SVZ ol the lateral ganglionic eminencie (LGE), the so-called basal progenitors. The cells positive for Nolz1 come from Gsh2 positive neural precursors, and its expression contributes to the inititation of a source of retinoic acid signalling within the striatum during development. Nolz1, in addition, regulates the Notch pathway in neural precursors an promotes the appearance of oligodendrocyte precursors in the LGE by a non-autonomous mechanism. On the other hand, we demostrate that the Ikaros isoform Ik1 is essential for the correct generation of the striatal enkephalinergic neurons from the differentiation of Dlx2 positive neural precursors of the LGE. We show that Ik1 is necessary for the cell-cycle exit of the neural precursors which will populate the striatum by the increase of the cyclin inhibitor p21. According with this observation, Ikaros absence during development specifically affects late striatal neurogenesis.This is translated in the adult Ikaros deficient animal in a reduced striatum due to a reduced number of matrix enkephalinergic projection neurons. Thus, Nolz1 and Ikaros exhert its actions during different times in striatal development, being Nolz1 role earlier and focused in the control of neural precursors, and Ikaros role occurs later, focused in the differentiation of the enkephalinergic projection neurons.

Este estudio ha tenido como objetivo principal la comprensión del proceso de aprendizaje de las ciencias experimentales en la educación infantil con niños de 5-6 años de edad. El proceso de aprendizaje se ha estudiado considerando las teorías del cambio conceptual y la construcción de modelos científicos precursores (MCP) en un contexto dialógico de instrucción. Para tal efecto, el estudio abordó dos estudios de casos: el fenómeno de flotación y hundimiento de los cuerpos y la caracterización de seres vivos. El MCP de flotación se caracterizó considerando propiedades relevantes de los objetos relacionadas con la densidad (peso, tamaño, hueco o lleno). EL MCP de ser vivo se caracterizó en base a propiedades biológicas tales como utilización de energía (nutrición, movimiento, excresión, respiración), crecimiento y desarrollo, y reproducción. Los estudios exploratorios se llevaron a cabo con un grupo de 25 niños en tres fases: pre-test, proceso de instrucción y post-test. El cambio conceptual se evaluó considerando las diferencias entre los pre-tests y los post-test en base a los modelos de Chi et al. (1994) y Thagard (1992), y la perspectiva contextual de Printich et al., (1993).Por otra parte, se analizaron las interacciones verbales para caracterizar los tipos de conversaciones que tuvieron lugar entre los niños y la docente durante el proceso de instrucción, y su incidencia en la promoción del cambio conceptual. Los resultados mostraron que los niños presentaron diferentes patrones de cambio conceptual dando lugar a diferentes patrones de aprendizaje y éstos, a su vez, a la construcción de los modelos científicos precursores de flotación y ser vivo propuestos. En el caso de flotación los cambios fueron principalmente epistemológicos en los cuales la mayoría de los niños enriquecieron su comprensión del fenómeno permitiéndoles construir el modelo precursor formulado. En el caso de seres vivos los cambios fueron epistemológicos y ontológicos con un enriquecimiento y una reestructuración de su comprensión, y de esta forma, también la mayoría de ellos construyeron el modelo precursor de ser vivo. Por otra parte, las interacciones verbales entre la docente y los niños mostraron evidencia de los diversos recursos que utilizaron los niños para enriquecer y reestructurar su conocimiento y que la mediación de la docente fue fundamental en este proceso. La perspectiva didáctica basada en la visión socioconstructivista del aprendizaje, la valoración del aprendizaje como cambio conceptual y la estrategia de instrucción basada en la modelización, mostró ser adecuada para comprender los procesos de aprendizaje en el aula. / This study had as main objective the understanding of experimental science learning in early childhood education with children aged 5-6 years old. The learning process has been studied considering the theories of conceptual change and the construction of scientific models precursors (MCPs) in a dialogic instruction. To that end, the study dealt with two case studies: the phenomenon of floating and sinking and the characterization of living things. MCP flotation considered relevant properties of objects related to the density (weight, size, hollow or full), and the MCP of living was based on biological properties such as utilization of energy (nutrition, movement, excretion, respiration), growth and development, and reproduction. Exploratory studies were conducted with a group of 25 children in three phases: pre-test, instructional process and post-test. Conceptual change was evaluated considering the differences between the pre-tests and post-test based on the model Chi et al. (1994) and Thagard (1992), and the contextual perspective of Printich et al. (1993). Moreover, verbal interactions were analyzed to characterize the types of conversations that occurred among children and teacher during the instructional process, and its impact on the promotion of conceptual change.Results showed that children had different patterns of conceptual change leading to different patterns of learning and, in turn, the construction of the scientific precursor models proposed. In the case of flotation, changes were mostly epistemological in which the majority of children enriched their understanding of the phenomenon by allowing the construction of the precursor model formulated. In the case of living things changes were epistemological and ontological leading children enrich and restructure their understanding, and so most of them also built the forerunner model of living thing. Moreover, the verbal interactions between teachers and children showed evidence of various resources used by children to enhance and restructure their knowledge and that the mediation of the teacher was crucial in this process. The educational perspective based on the socio-constructivist perspective, assessment of learning as conceptual change and strategy-based instruction in modelling, showed to be adequate for understanding the processes of learning in the classroom.

<p>Development requires a delicate balance of proliferation and differentiation. Too little proliferation can result in dysfunctional tissues, while prolonged or heightened proliferation can result in tumor formation. This is clearly seen with the granule neuron precursors (GNPs) of the cerebellum. Too little proliferation of these cells during development results in ataxia, whereas too much proliferation results in the cerebellar tumor medulloblastoma. While these cells are known to proliferate in response to Shh, it is not clear what controls the differentiation of these cells in vivo.</p><p> Previous work from our lab has identified basic fibroblast growth factor (bFGF) as a candidate differentiation factor for these cells. In this thesis, I characterize some of the cellular and molecular mechanisms involved in FGF-mediated inhibition (FMI) of Shh-induced GNP proliferation. In addition, I employ FGFR knockouts and a bFGF gain-of-function mouse to determine whether FGF signaling is necessary and/or sufficient for differentiation of GNPs during cerebellar development. Finally, the question of whether bFGF can be effective as a therapeutic agent for in vivo tumor treatment is tested in a transplant model.</p><p> These experiments indicate that FGF signaling is neither necessary nor sufficient for GNP differentiation during cerebellar development. However, transplanted tumors are potently inhibited by bFGF treatment. Furthermore, FMI is shown to occur around the level of Gli2 processing in the Shh pathway, implying that such a treatment has promise to be widely effective in treatment of Shh-dependent medulloblastomas.</p> / Dissertation

Combustor blowout is a very serious concern in modern land-based and aircraft engine combustors. The ability to sense blowout precursors can provide significant payoffs in engine reliability and life. The objective of this work is to characterize the blowout phenomenon and develop a sensing methodology which can detect and assess the proximity of a combustor to blowout by monitoring its acoustic signature, thus providing early warning before the actual blowout of the combustor. The first part of the work examines the blowout phenomenon in a piloted jet burner. As blowout was approached, the flame detached from one side of the burner and showed increased flame tip fluctuations, resulting in an increase in low frequency acoustics. Work was then focused on swirling combustion systems. Close to blowout, localized extinction/re-ignition events were observed, which manifested as bursts in the acoustic signal. These events increased in number and duration as the combustor approached blowout, resulting an increase in low frequency acoustics. A variety of spectral, wavelet and thresholding based approaches were developed to detect precursors to blowout. The third part of the study focused on a bluff body burner. It characterized the underlying flame dynamics near blowout in greater detail and related it to the observed acoustic emissions. Vorticity was found to play a significant role in the flame dynamics. The flame passed through two distinct stages prior to blowout. The first was associated with momentary strain levels that exceed the flames extinction strain rate, leading to flame holes. The second was due to large scale alteration of the fluid dynamics in the bluff body wake, leading to violent flapping of the flame front and even larger straining of the flame. This led to low frequency acoustic oscillations, of the order of von Karman vortex shedding. This manifested as an abrupt increase in combustion noise spectra at 40-100 Hz very close to blowout. Finally, work was also done to improve the robustness of lean blowout detection by developing integration techniques that combined data from acoustic and optical sensors.