Nutritional control of mRNA isoform expression during developmental arrest and recovery in C. elegans.

Nutrient availability profoundly influences gene expression. Many animal genes encode multiple transcript isoforms, yet the effect of nutrient availability on transcript isoform expression has not been studied in genome-wide fashion. When Caenorhabditis elegans larvae hatch without food, they arrest development in the first larval stage (L1 arrest). Starved larvae can survive L1 arrest for weeks, but growth and post-embryonic development are rapidly initiated in response to feeding. We used RNA-seq to characterize the transcriptome during L1 arrest and over time after feeding. Twenty-seven percent of detectable protein-coding genes were differentially expressed during recovery from L1 arrest, with the majority of changes initiating within the first hour, demonstrating widespread, acute effects of nutrient availability on gene expression. We used two independent approaches to track expression of individual exons and mRNA isoforms, and we connected changes in expression to functional consequences by mining a variety of databases. These two approaches identified an overlapping set of genes with alternative isoform expression, and they converged on common functional patterns. Genes affecting mRNA splicing and translation are regulated by alternative isoform expression, revealing post-transcriptional consequences of nutrient availability on gene regulation. We also found that phosphorylation sites are often alternatively expressed, revealing a common mode by which alternative isoform expression modifies protein function and signal transduction. Our results detail rich changes in C. elegans gene expression as larvae initiate growth and post-embryonic development, and they provide an excellent resource for ongoing investigation of transcriptional regulation and developmental physiology.

Later Life Consequences of Subteratogenic Exposure to a Complex PAH Mixture in the Atlantic Killifish (Fundulus heteroclitus)

Subteratogenic and other low-level chronic exposures to toxicant mixtures are an understudied threat to environmental and human health. It is especially important to understand the effects of these exposures for contaminants, such as polycyclic aromatic hydrocarbons (PAHs) a large group of more than 100 individual compounds, which are important environmental (including aquatic) contaminants. Aquatic sediments constitute a major sink for hydrophobic pollutants, and studies show PAHs can persist in sediments over time. Furthermore, estuarine systems (namely breeding grounds) are of particular concern, as they are highly impacted by a wide variety of pollutants, and estuarine fishes are often exposed to some of the highest levels of contaminants of any vertebrate taxon. Acute embryonic exposure to PAHs results in cardiac teratogenesis in fish, and early life exposure to certain individual PAHs and PAH mixtures cause heart alterations with decreased swimming capacity in adult fish. Consequently, the heart and cardiorespiratory system are thought to be targets of PAH mixture exposure. While many studies have investigated acute, teratogenic PAH exposures, few studies have longitudinally examined the impacts of subtle, subteratogenic PAH mixture exposures, which are arguably more broadly applicable to environmental contamination scenarios. The goal of this dissertation was to highlight the later-life consequences of early-life exposure to subteratogenic concentrations of a complex, environmentally relevant PAH mixture.

A unique population of Fundulus heteroclitus (the Atlantic killifish or mummichog, hereafter referred to as killifish), has adapted to creosote-based polycyclic aromatic hydrocarbons (PAHs) found at the Atlantic Wood Industries (AW) Superfund site in the southern branch of the Elizabeth River, VA, USA. This killifish population survives in a site heavily contaminated with a mixture of PAHs from former creosote operations. They have developed resistance to the acute toxicity and teratogenic effects caused by the mixture of PAHs in sediment from the site. The primary goal of this dissertation was to compare and contrast later-life outcomes of early-life, subteratogenic PAH mixture exposure in both the Atlantic Wood killifish (AW) and a naïve reference population of killifish from King’s Creek (KC; a relatively uncontaminated tributary of the Severn River, VA). Killifish from both populations were exposed to subteratogenic concentrations of a complex PAH-sediment extract, Elizabeth River Sediment Extract (ERSE), made by collecting sediment from the AW site. Fish were reared over a 5-month period in the laboratory, during which they were examined for a variety of molecular, physiological and behavioral responses.

The central aims of my dissertation were to determine alterations to embryonic gene expression, larval swimming activity, adult behavior, heart structure, enzyme activity, and swimming/cardiorespiratory performance following subteratogenic exposure to ERSE. I hypothesized that subteratogenic exposure to ERSE would impair cardiac ontogenic processes in a way that would be detectable via gene expression in embryos, and that the misregulation of cardiac genes would help to explain activity changes, behavioral deficits, and later-life swimming deficiencies. I also hypothesized that fish heart structure would be altered. In addition, I hypothesized that the AW killifish population would be resistant to developmental exposures and perform normally in later life challenges. To investigate these hypotheses, a series of experiments were carried out in PAH-adapted killifish from Elizabeth River and in reference killifish. As an ancillary project to the primary aims of the dissertation, I examined the toxicity of weaker aryl hydrocarbon receptor (AHR) agonists in combination with fluoranthene (FL), an inhibitor of cytochrome P4501A1 (CYP1A1). This side project was conducted in both Danio rerio (zebrafish) and the KC and AW killifish.

Embryonic gene expression was measured in both killifish populations over an ERSE dose response with multiple time points (12, 24, 48, and 144 hours post exposure). Genes known to play critical roles in cardiac structure/development, cardiac function, and angiogenesis were elevated, indicating cardiac damage and activation of cardiovascular repair mechanisms. These data helped to inform later-life swimming performance and cardiac histology studies. Behavior was assessed during light and dark cycles in larvae of both populations following developmental exposure to ERSE. While KC killifish showed activity differences following exposure, AW killifish showed no significant changes even at concentrations that would cause overt cardiac toxicity in KC killifish. Juvenile behavior experiments demonstrated hyperactivity following ERSE exposure in KC killifish, but no significant behavioral changes in AW killifish. Adult swimming performance via prolonged critical swimming capacity (Ucrit) demonstrated performance costs in the AW killifish. Furthermore, swimming performance decline was observed in KC killifish following exposure to increasing dilutions of ERSE. Lastly, cardiac histology suggested that early-life exposure to ERSE could result in cardiac structural alteration and extravasation of blood into the pericardial cavity.

Responses to AHR agonists resulted in a ranking of relative potency for agonists, and determined which agonists, when combined with FL, caused cardiac teratogenesis. These experiments showed interesting species differences for zebrafish and killifish. To probe mechanisms responsible for cardiotoxicity, a CYP1A-morpholino and a AHR2-morpholino were used to mimic FL effects or attempt to rescue cardiac deformities respectively. Findings suggested that the cardiac toxicity elicited by weak agonist + FL exposure was likely driven by AHR-independent mechanisms. These studies stand in contrast to previous research from our lab showing that moderate AHR agonist + FL caused cardiac toxicity that can be partially rescued by AHR-morpholino knockdown.

My findings will form better characterization of mechanisms of PAH toxicity, and advance our understanding of how subteratogenic mixtures of PAHs exert their toxic action in naïve killifish. Furthermore, these studies will provide a framework for investigating how subteratogenic exposures to PAH mixtures can impact aquatic organismal health and performance. Most importantly, these experiments have the potential to help inform risk assessment in fish, mammals, and potentially humans. Ultimately, this research will help protect populations exposed to subtle PAH-contamination.

Global value chains in a post-Washington Consensus world

Contemporary globalization has been marked by significant shifts in the organization and governance of global industries. In the 1970s and 1980s, one such shift was characterized by the emergence of buyer-driven and producer-driven commodity chains. In the early 2000s, a more differentiated typology of governance structures was introduced, which focused on new types of coordination in global value chains (GVCs). Today the organization of the global economy is entering another phase, with transformations that are reshaping the governance structures of both GVCs and global capitalism at various levels: (1) the end of the Washington Consensus and the rise of contending centers of economic and political power; (2) a combination of geographic consolidation and value chain concentration in the global supply base, which, in some cases, is shifting bargaining power from lead firms in GVCs to large suppliers in developing economies; (3) new patterns of strategic coordination among value chain actors; (4) a shift in the end markets of many GVCs accelerated by the economic crisis of 2008-09, which is redefining regional geographies of investment and trade; and (5) a diffusion of the GVC approach to major international donor agencies, which is prompting a reformulation of established development paradigms. © 2013 © 2013 Taylor & Francis.

Post-1945 Danish Piano Trio Literature: A Recording Dissertation

This recording dissertation surveys post-1945 literature written for piano trio (violin, violoncello and piano) by ten Danish composers. The literature was first considered for inclusion by searching a database provided by the Danish Music Information Center (www.mic.dk). Scores were rented from the publisher Edition Wilhelm Hansen AS, or purchased from the publisher Samfundet til Udgivelse af Dansk Musik. An additional score published by Viking Musikforlag was used as well. The music was then studied and evaluated for selection. During the selection process, the following criteria were considered: 1) quality of the compositions; 2) recognition of the composers at the national or international level; 3) whether the compositions had been previously recorded; and 4) variety of compositional styles. The selected works are written by Niels Viggo Bentzon, Vagn Holmboe, Anders Koppel, Herman D. Koppel, Bent Lorentzen, Anders Nordentoft, Per Norgard, Michael Nyvang, Karl Aage Rasmussen, and Poul Rovsing Olsen. The selected compositions were practiced, rehearsed, and performed under direct supervision of the composers and other expert musicians. In order to better understand the compositional style of each composer, relevant books, articles, and recordings were researched and studied. This recording dissertation is supported by a written document. A subjective preference for program balance was exercised to determine the order of recorded works. The written document is divided into chapters defined by composer, following the order of the recorded document, which include the composers' biographies and notes referring to the recorded compositions. The recording took place at the Manzius Gaarden, Birkerod, Denmark during three sessions: July 31-August 2, 2002, March 2 and 3, 2003, and June 2-4, 2003. The music for this dissertation was recorded by the members of the Jalina Trio; Line Fredens, violin, Janne Fredens, cello and Natsuki Fukasawa, piano. Aksel Trige, a well-respected recording engineer, was engaged for the recording and editing. Additionally, a Hamburg Steinway concert grand piano was rented and a Joseph Guarnerius filius Andreas Cremona violin (1706) was provided by the Augustinus Fonden, Denmark. The cellist used her own instrument, Vuillaume of Paris (c. 1850). The expense of this recording was partially paid by generous grants from the Augustinus Fonden, the Solist Foreningen af 1921, and the Dansk Musikerforbunds Kollective Rettighedsmidler. The compositions selected for this recording dissertation are assumed to be previously unrecorded, with the exception of Poul Rovsing Olsen's Trio II.

Sea Urchin Body Plan Development and Evolution: An Integrative Transcriptomic Approach

My dissertation work integrates comparative transcriptomics and functional analyses to investigate gene expression changes underlying two significant aspects of sea urchin evolution and development: the dramatic developmental changes associated with an ecologically significant shift in life history strategy and the development of the unusual radial body plan of adult sea urchins.

In Chapter 2, I investigate evolutionary changes in gene expression underlying the switch from feeding (planktotrophic) to nonfeeding (lecithotrophic) development in sea urchins. In order to identify these changes, I used Illumina RNA-seq to measure expression dynamics across 7 developmental stages in three sea urchin species: the lecithotroph Heliocidaris erythrogramma, the closely related planktotroph Heliocidaris tuberculata, and an outgroup planktotroph Lytechinus variegatus. My analyses draw on a well-characterized developmental gene regulatory network (GRN) in sea urchins to understand how the ancestral planktotrophic developmental program was altered during the evolution of lecithotrophic development. My results suggest that changes in gene expression profiles occurred more frequently across the transcriptome during the evolution of lecithotrophy than during the persistence of planktotrophy. These changes were even more pronounced within the GRN than across the transcriptome as a whole, and occurred in each network territory (skeletogenic, endomesoderm and ectoderm). I found evidence for both conservation and divergence of regulatory interactions in the network, as well as significant changes in the expression of genes with known roles in larval skeletogenesis, which is dramatically altered in lecithotrophs. I further explored network dynamics between species using coexpression analyses, which allowed me to identify novel players likely involved in sea urchin neurogenesis and endoderm patterning.

In Chapter 3, I investigate developmental changes in gene expression underlying radial body plan development and metamorphosis in H. erythrogramma. Using Illumina RNA-seq, I measured gene expression profiles across larval, metamorphic, and post-metamorphic life cycle phases. My results present a high-resolution view of gene expression dynamics during the complex transition from pre- to post-metamorphic development and suggest that distinct sets of regulatory and effector proteins are used during different life history phases.

Collectively, my investigations provide an important foundation for future, empirical studies to investigate the functional role of gene expression change in the evolution of developmental differences between species and also for the generation of the unusual radial body plan of sea urchins.

MiR-215 Is Induced Post-transcriptionally via HIF-Drosha Complex and Mediates Glioma-Initiating Cell Adaptation to Hypoxia by Targeting KDM1B.

The hypoxic tumor microenvironment serves as a niche for maintaining the glioma-initiating cells (GICs) that are critical for glioblastoma (GBM) occurrence and recurrence. Here, we report that hypoxia-induced miR-215 is vital for reprograming GICs to fit the hypoxic microenvironment via suppressing the expression of an epigenetic regulator KDM1B and modulating activities of multiple pathways. Interestingly, biogenesis of miR-215 and several miRNAs is accelerated post-transcriptionally by hypoxia-inducible factors (HIFs) through HIF-Drosha interaction. Moreover, miR-215 expression correlates inversely with KDM1B while correlating positively with HIF1α and GBM progression in patients. These findings reveal a direct role of HIF in regulating miRNA biogenesis and consequently activating the miR-215-KDM1B-mediated signaling required for GIC adaptation to hypoxia.

Toward understanding the essence of post-translational modifications for the Mycobacterium tuberculosis immunoproteome

CD4⁺ T cells are prominent effector cells in controlling Mycobacterium tuberculosis (Mtb) infection but may also contribute to immunopathology. Studies probing the CD4⁺ T cell response from individuals latently infected with Mtb or patients with active tuberculosis using either small or proteome-wide antigen screens so far revealed a multi-antigenic, yet mostly invariable repertoire of immunogenic Mtb proteins. Recent developments in mass spectrometry-based proteomics have highlighted the occurrence of numerous types of post-translational modifications (PTMs) in proteomes of prokaryotes, including Mtb. The well-known PTMs in Mtb are glycosylation, lipidation, or phosphorylation, known regulators of protein function or compartmentalization. Other PTMs include methylation, acetylation, and pupylation, involved in protein stability. While all PTMs add variability to the Mtb proteome, relatively little is understood about their role in the anti-Mtb immune responses. Here,we reviewMtb protein PTMs and methods to assess their role in protective immunity against Mtb. © 2014 van Els, Corbière, Smits, vanGaans-van den Brink, Poelen, Mascart, Meiring and Locht.

Factores ambientales pre - dispersión y post - dispersión de semillas que modulan la germinación y emergencia de Digitaria sanguinalis (L.) Scop. en el cultivo de soja

El éxito de Digitaria sanguinalis en los cultivos estivales de la Argentina se debe en gran medida al establecimiento de varias cohortes a lo largo del ciclo del cultivo, lo que le permite escapar a los controles químicos. El establecimiento de las plántulas depende de factores ambientales pre-dispersión (competitivos y no competitivos), a través de los efectos maternos, y post-dispersión (no competitivos). A su vez, los efectos predispersión competitivos también afectan la fecundidad de las plantas. Por lo tanto, comprender los efectos de las interacciones dentro del sistema cultivo-maleza sería de utilidad para diseñar estrategias de manejo más efectivas de la maleza. Este trabajo tuvo como objetivo general i) determinar los efectos pre-dispersión que genera el cultivo de soja sobre el crecimiento, la estructura, la fecundidad y la dormición de semillas de biotipos locales de D. sanguinalis, y ii) los efectos post-dispersión que genera el ambiente en el que se encuentran las semillas condicionando el establecimiento de las plántulas en la campaña siguiente. Para cumplir con estos objetivos se realizaron experimentos i) en cámara evaluando los efectos de la temperatura, luz y humedad y el rol de las cubiertas en la imposición de la dormición, ii) en parcelas a campo combinando la presencia del cultivo con distintas distancias entre surcos, los grupos de madurez y los "gaps", iii) en macetas a campo evaluando los efectos del sombreo, la fertilidad edáfica y la luz roja y azul y iv) en parcelas a campo combinando la cobertura del suelo (rastrojo), el tiempo de permanencia de las semillas en el suelo, la presencia del cultivo de soja y el nivel de dormición de las semillas al momento de su dispersión. Temperaturas frescas (5ºC a 20ºC) y humedad, seguidas de temperaturas alternadas (20/30ºC) con luz fueron las mejores condiciones para la salida y la terminación de la dormición de las semillas, respectivamente. Dicha dormición está determinada por las cubiertas de las semillas, principalmente por la lemma y, aparentemente por el efecto de inhibidores presentes en ella. El cultivo de soja modificó el ambiente en el que crecen las plantas de la maleza al alterar la radiación incidente, la relación R-RL, la temperatura y la humedad relativa reduciendo la biomasa, la altura, el número de vástagos, la fecundidad de las plantas y el nivel de dormición de las semillas de D. sanguinalis. El nivel de dormición fue afectado por la temperatura máxima, la alternancia de temperatura y la radiación incidente en estadios reproductivos de la maleza, en cambio la fertilidad edáfica, la luz azul y rojo lejano no tuvieron ningún efecto. Por otra parte, el rastrojo de soja o de maíz, no modificó la salida de la dormición pero retrasó la terminación de la misma por la disminución en la alternancia de las temperaturas, retrasando la germinación y la emergencia de la maleza en la campaña siguiente. Semillas con distinto nivel de dormición en el momento de dispersión, tuvieron distintos valores de emergencia en la campaña siguiente cuando la temperatura no fue lo suficientemente baja como para reducir los niveles de dormición de toda la población. La presencia de un cultivo de soja junto con la presencia de rastrojo de soja o maíz, redujeron la emergencia de las plántulas a campo debido, en parte, al efecto de la menor alternancia de temperaturas. Los resultados de esta tesis permiten determinar cómo la estructura del cultivo de soja modifica el establecimiento de plántulas de D. sanguinalis en la campaña siguiente, a través de su efecto sobre el ambiente pre y postdispersión, afectando la salida y la terminación de la dormición y por lo tanto la emergencia de la maleza.

El estrés térmico post - floración y sus efectos sobre el peso y la calidad de los granos en maíces de diferente destino de producción

En maíz, los golpes de calor (días con temperaturas menor a 35 grados C) ocurridos tempranamente durante el período post-floración pueden afectar negativamente el peso individual de los granos, a traves de limitaciones del crecimiento del cultivo. El conocimiento que existe respecto a las bases eco-fisiológicas de la respuesta de la calidad de los granos a la incidencia de este estres durante el llenado efectivo es prácticamente nulo. El objetivo de este trabajo fue analizar en condiciones a campo el impacto de los golpes de calor durante las etapas tempranas y tardías del llenado efectivo sobre el peso, la calidad general y específica de la industria de los granos en híbridos de maíz de diferente destino de producción (colorado duro o flint, pisingallo y semi-dentad). Los golpes de calor, especialmente aquellos ocurridos durante las etapas tempranas del llenado, redujeron el crecimiento de los cultivos por caídas de la eficiencia en el uso de la radiacion interceptada. Esto produjo un desbalance entre la disponibilidad de asimilados y el requerimiento de estos por parte de los granos que se tradujo en una interrupción prematura del llenado. Consecuentemente, se obtuvieron granos mas livianos con concentraciones mas bajas de aceite y mas altas de proteína. Estas respuestas fueron acompanadas de alteraciones en la composición específica de las proteínas de reserva, incrementándose la abundancia relativa de glutelinas, y ƒÀ- y ƒÁ- zeinas, en detrimento de aquella de ƒ¿-zeinas. Asimismo, los golpes de calor provocaron efectos negativos sobre los diversos parámetros de calidad industrial de los granos, asociados principalmente a disminuciones en la abundancia relativa de ƒ¿-zeínas. Para la mayoría de los rasgos mencionados, los híbridos semi-dentados tuvieron una respuesta mas pronunciada al estrés que los híbridos colorado duro y pisingallo, aparentando ser sensibles a temperaturas del aire incluso inferiores a 35 grados C.

Contenido de nitrógeno en grano de trigo y cebada cervecera ante condiciones contrastantes de fuente - destino en post - antesis

Trigo y cebada son cultivos con un alto grado de difusión a nivel mundial. Ambos son importantes como especies cultivables debido a la utilidad que poseen sus granos. El principal destino de trigo es la industria harinera y el de cebada la industria maltera. Estas industrias requieren que el grano posea un porcentaje de proteína determinado. El objetivo general del trabajo fue analizar en forma comparada para trigo y cebada las variaciones en el porcentaje de nitrógeno en el grano ante modificaciones en la relación fuente - destino en post-antesis. Los factores considerados en el análisis fueron (i) un cultivar de trigo y cebada, (ii) dos niveles contrastantes de disponibilidades de nitrógeno en suelo a siembra (57 kg N ha-1, N0; 130 kg N ha-1, N1), (iii) variaciones en la relación fuente - destino (F:D) a lo largo del período de llenado (7, 14, 21 y 28 días desde antesis), dadas por (iii.1) sin modificación en la relación fuente - destino (control), (iii.2) incremento en la disponibilidad de recursos por grano mediante desespiguillado (tratamiento DE), (iii.3) disminución parcial en el tamaño de la fuente foliar actual (defoliación de todas las hojas verdes del cultivo con excepción de la hoja bandera, tratamiento HB), y (iii.4) supresión total en el tamaño de la fuente foliar actual (defoliación de todas las hojas verdes, tratamiento SH). El desespiguillado en trigo aumentó el porcentaje de nitrógeno en grano (PNG) independientemente del momento en el cual se realizó dicho tratamiento, y con mayor efecto en la situación NO que N1. En cebada, solo el tratamiento DE 7 generó incrementos en el PNG respecto al control. Por consiguiente, la limitación por nitrógeno post-antesis fue mayor en trigo que en cebada y se evidenció hasta estados intermedio - avanzados del período de llenado. Los tratamientos de defoliaciones parciales o totales no presentaron diferencias en PNG con respecto a la situación control en ningún momento del periodo de llenado en el que se realizó el tratamiento en ninguno de los cultivares. La respuesta del PNG a los cambios F:D de granos provenientes del vástago principal fue semejante a la de los granos de los macollos.

El estrés térmico post - floración y sus efectos sobre el peso y la calidad de los granos en maíces de diferente destino de producción

En maíz, los golpes de calor (días con temperaturas menor a 35 grados C) ocurridos tempranamente durante el período post-floración pueden afectar negativamente el peso individual de los granos, a traves de limitaciones del crecimiento del cultivo. El conocimiento que existe respecto a las bases eco-fisiológicas de la respuesta de la calidad de los granos a la incidencia de este estres durante el llenado efectivo es prácticamente nulo. El objetivo de este trabajo fue analizar en condiciones a campo el impacto de los golpes de calor durante las etapas tempranas y tardías del llenado efectivo sobre el peso, la calidad general y específica de la industria de los granos en híbridos de maíz de diferente destino de producción (colorado duro o flint, pisingallo y semi-dentad). Los golpes de calor, especialmente aquellos ocurridos durante las etapas tempranas del llenado, redujeron el crecimiento de los cultivos por caídas de la eficiencia en el uso de la radiacion interceptada. Esto produjo un desbalance entre la disponibilidad de asimilados y el requerimiento de estos por parte de los granos que se tradujo en una interrupción prematura del llenado. Consecuentemente, se obtuvieron granos mas livianos con concentraciones mas bajas de aceite y mas altas de proteína. Estas respuestas fueron acompanadas de alteraciones en la composición específica de las proteínas de reserva, incrementándose la abundancia relativa de glutelinas, y ƒÀ- y ƒÁ- zeinas, en detrimento de aquella de ƒ¿-zeinas. Asimismo, los golpes de calor provocaron efectos negativos sobre los diversos parámetros de calidad industrial de los granos, asociados principalmente a disminuciones en la abundancia relativa de ƒ¿-zeínas. Para la mayoría de los rasgos mencionados, los híbridos semi-dentados tuvieron una respuesta mas pronunciada al estrés que los híbridos colorado duro y pisingallo, aparentando ser sensibles a temperaturas del aire incluso inferiores a 35 grados C.

Distributed digital post-production

It is now possible to use powerful general purpose computer architectures to support post-production of both video and multimedia projects. By devising a suitable portable software architecture and using high-speed networking in an appropriate manner, a system has been constructed where editors are no longer tied to a specific location. New types of production, such as multi-threaded interactive video, are supported. Editors may also work remotely where very high speed network connection is not currently provided. An object-oriented database is used for the comprehensive cataloging of material and to support automatic audio/video object migration and replication. Copyright © 1997 by the Society of Motion Picture and Television Engineers, Inc.

A computational study of the characteristics of aircraft post-crash fires

Full-scale furnished cabin fires have been studied experimentally for the purpose of characterising the post-crash cabin fire environment by the US Federal Aviation Administration for many years. In this paper the Computational Fluid Dynamics fire field model SMARTFIRE is used to simulate one of these fires conducted in the C-133 test facility in order to provide further validation of the computational approach and the SMARTFIRE software. The experiment involves exposing the interior cabin materials to an external fuel fire, opening only one exit at the far end of the cabin (the same side as the rupture) for ventilation, and noting the subsequent spread of the external fire to the cabin interior and the onset of flashover at approximately 210 seconds. Through this analysis, the software is shown to be in good agreement with the experimental data, producing reasonable agreement with the fire dynamics prior to flashover and producing a reasonable prediction of the flashover time i.e. 225 seconds. The paper then proceeds to utilize the model to examine the impact on flashover time of the extent of cabin furnishings and cabin ventilation provided by available exits