Exploring Saccharomyces cerevisiae’s responses to acetic acid and other inhibitors found in lignocellulosic hydrolysates

The limited tolerance of the budding yeast Saccharomyces cerevisiae to lignocellulosic hydrolysate inhibitors is a key challenge to its use in biorefinery cell factories. Considerable resources have been invested in the isolation of yeast strains with better tolerance towards the inhibitors released during lignocellulose hydrolysis, such as acetic acid. The goal of this thesis was twofold: characterize the transcriptional response of S. cerevisiae to wheat straw hydrolysate and explore the role of essential S. cerevisiae genes in acetic and formic acid tolerance, using a new biosensor and competitive growth assays.The transcriptomes of one laboratory strain, two industrial strains, and two wild-type isolates grown in wheat straw hydrolysate were profiled. Despite similar growth, the isolates showed different expression of genes encoding proteins involved in oxidative stress response, lipid accumulation, and transport, suggesting different genetic strategies for tolerance. The new acetic acid biosensor was based on two transcription factors, Haa1 from S. cerevisiae and BM3R1 from Bacillus megaterium. Biosensor and competitive growth were used in parallel to screen a S. cerevisiae CRISPR interference strain library. While fluorescence-activated cell sorting led to the isolation of cells with higher acetic acid retention and sensitivity, competitive growth assays allowed the identification of cells with higher acid tolerance. The results confirmed the role in acid stress response of genes involved in glycogen accumulation, chromatin modification, and mitochondrial or proteasomal functions. Two novel targets for improving tolerance were also identified: PAP1 and HIP1. Altogether, this thesis provides mechanistic insight into the stress response to lignocellulosic hydrolysates or weak acid inhibitors that limit yeast-mediated conversion of lignocellulosic biomass into biochemicals. Additionally, it offers new tools for the identification of strains with altered acetic acid tolerance

Intercultural Victorians: the challenge of modern South Korean Protestant mission

The modern South Korean Protestant Church, widely seen as the second largest sender of Christian missionaries in the world, has reached an important point in its development as a global missionary power. While there has been rapid growth in missionary numbers and some encouraging results, critics of South Korea mission (including those within and outside of the movement) are more aware than ever of the problems facing Korean missionaries on the field in areas like cultural adaptation and building effective relationships. This thesis proposes that South Korean mission can be analysed through two elements that are clearly seen through written and oral sources: the "Victorian" and the "Intercultural."On the "Victorian" side, clear similarities connect Korean mission and that of the West, especially the United States, in the late nineteenth and early twentieth centuries. This resemblance can be seen in several facets. Imbued with a high sense of purpose and ambition, Korean missionaries are unencumbered by historical baggage. They also face some of the same difficulties in areas like cultural adaptation and the use of money. Moreover, some view Korean missionaries as heirs to the "Golden Age" of mission who can complete the task of evangelisation.By "Intercultural," I am referring both to the way that the word epitomises a sense of giving and taking within mission and the fact that it is currently the most fashionable of the terms used for cultural adaptation within mission. Furthermore, the term ties into the hope of Korean mission to act as an intermediary between Western and non-Western cultures. Aware of their problems with cultural adaptation, Korean missionaries are attempting to embrace the newest ideas about culture in mission, particularly through training. However, much remains to be done before the desired deeper relationships and reciprocity that interculturation suggests are brought about within Korean mission. The challenge for the Korean Church is to integrate the intercultural ideals of modern mission while maintaining the enthusiasm and purpose that has drive Korean mission forward

Sunbelt Dreams and Altered States: A Social and Cultural History of Florida, 1950-2000

Florida\u27s Big Bang represents an astonishing and complex story, a state swelling from 2.7 million inhabitannts in 1950 to 16 million only fifty years later. Demographically, Florida\u27s tranformation is nothing short of revolutionary. On the eve of World War II Florida\u27s population of l.9 million ranked twenty-seventh nationally and last in the South, trailing even lowly South Carolina and Arkansas. America\u27s twentieth most populous state in 1950, Florida has vaulted to America\u27s fourth largest in 2000, and stands poised to overtake New York. Like shifting tectonic plates, the post-World War II decade witnessed one of the great population shift in history. Million of Americans residing in the North and Midwest migrated to the South and West. Millions of emigrants from the Caribbean, Latin America, and Asia flocked to California, Texas, and Florida. Between 1970 and 1990, as America\u27s population grew by 21 percent, the South surged by 40 percent, while Florida soared by 76 percent. In the last half century, while California and Texas tripled their populations, Florida advanced six-fold.

GI Joe Meets Jim Crow: Racial Violence and Reform in World War II Florida

Where were you on December 7, 1941, and what did you experience on that memorable day? If you were Master Sergeant Warren Bryant, stationed at Tampa’s MacDill Field, you were reminded of your place in American society. Bryant explained the coming of war: “When the Japanese bombed Pearl Harbor . . . all of the whites at MacDill Field were running around with loaded guns. We [blacks] had no guns and no idea of what was going on, so you can imagine what was running through our minds until we learned of the Japanese attack. Even with this knowledge it was of no comfort to be practically penned in our area with armed patty boys all over everywhere. We trusted them just about as much as a coiled rattlesnake.“1 “GI Joe Meets Jim Crow” examines the tensions and violence that erupted on and off military camps in Florida during World War II and their role in the development of the civil rights movement

Neuroanatomical spread of amyloid β and tau in Alzheimer's disease: implications for primary prevention.

With recent advances in our understanding of the continuous pathophysiological changes that begin many years prior to symptom onset, it is now apparent that Alzheimer's disease cannot be adequately described by discrete clinical stages, but should also incorporate the continuum of biological changes that precede and underlie the clinical representation of the disease. By jointly considering longitudinal changes of all available biomarkers and clinical assessments, variation within individuals can be integrated into a single continuous measure of disease progression and used to identify the earliest pathophysiological changes. Disease time, a measure of disease severity, was estimated using a Bayesian latent time joint mixed-effects model applied to an array of imaging, biomarker and neuropsychological data. Trajectories of regional amyloid β and tau PET uptake were estimated as a function of disease time. Regions with early signs of elevated amyloid β uptake were used to form an early, focal composite and compared to a commonly used global composite, in a separate validation sample. Disease time was estimated in 279 participants (183 cognitively unimpaired individuals, 61 mild cognitive impairment and 35 Alzheimer's disease dementia patients) with available amyloid β and tau PET data. Amyloid β PET uptake levels in the posterior cingulate and precuneus start high and immediately increase with small increases of disease time. Early elevation in tau PET uptake was found in the inferior temporal lobe, amygdala, banks of the superior temporal sulcus, entorhinal cortex, middle temporal lobe, inferior parietal lobe and the fusiform gyrus. In a separate validation sample of 188 cognitively unimpaired individuals, the early, focal amyloid β PET composite showed a 120% increase in the accumulation rate of amyloid β compared to the global composite (P < 0.001), resulting in a 60% increase in the power to detect a treatment effect in a primary prevention trial design. Ordering participants on a continuous disease time scale facilitates the inspection of the earliest signs of amyloid β and tau pathology. To detect early changes in amyloid β pathology, focusing on the earliest sites of amyloid β accumulation results in more powerful and efficient study designs in early Alzheimer's disease. Targeted composites could be used to re-examine the thresholds for amyloid β-related study inclusion, especially as the field shifts to focus on primary and secondary prevention. Clinical trials of anti-amyloid β treatments may benefit from the use of focal composites when estimating drug effects on amyloid β and tau changes in populations with minimal amyloid β and tau pathology and limited expected short-term accumulation

Development of an Haa1-based biosensor for acetic acid sensing in Saccharomyces cerevisiae

Acetic acid is one of the main inhibitors of lignocellulosic hydrolysates and acetic acid tolerance is crucial for the development of robust cell factories for conversion of biomass. As a precursor of acetyl-coenzyme A, it also plays an important role in central carbon metabolism. Thus, monitoring acetic acid levels is a crucial aspect when cultivating yeast. Transcription factor-based biosensors represent useful tools to follow metabolite concentrations. Here, we present the development of an acetic acid biosensor based on the Saccharomyces cerevisiae transcription factor Haa1 that upon binding to acetic acid relocates to the nucleus. In the biosensor, a synthetic transcription factor consisting of Haa1 and BM3R1 from Bacillus megaterium was used to control expression of a reporter gene under a promoter containing BM3R1 binding sites. The biosensor did not drive expression under a promoter containing Haa1 binding sites and responded to acetic acid over a linear range spanning from 10 to 60 mM. To validate its applicability, the biosensor was integrated into acetic acid-producing strains. A direct correlation between biosensor output and acetic acid production was detected. The developed biosensor enables high-throughput screening of strains producing acetic acid and could also be used to investigate acetic acid-tolerant strain libraries

LA CONVOCAZIONE DELLE SEDI PATRIARCALI D’ORIENTE ALL’VIII CONCILIO ECUMENICO (869-70): ANTIOCHIA E GERUSALEMME

L’ottavo concilio ecumenico, tenutosi nell’869-870, rappresentò un momento di svolta (seppur temporanea) nel decennale confronto tra le Chiese di Roma e Costantinopoli. Al contempo esso fu una occasione unica per il nuovo imperatore Basilio I di rivedere i rapporti tra l’impero ed il papato e ricostituire l’unità perduta dalla Chiesa bizantina. Parte integrante di questo suo progetto furono anche i restanti troni patriarcali d’Oriente, sottoposti al califfato abbaside. Essi furono direttamente coinvolti dall’imperatore nel proprio progetto di pacificazione ecclesiastica. Questo contributo ha analizzato le reazioni delle due Sedi che, per prime, inviarono i propri rappresentati al concilio: Antiochia e Gerusalemme. The eighth ecumenical council, held in 869-870, represented a turning point (even though temporary) in the decade-long confrontation between the Churches of Rome and Constantinople. At the same time the council was a unique opportunity for the new emperor Basil I to revise the relationship between the empire and the papacy and to reestablish the unity lost by the byzantine Church. A completing part of his project was also the remaining patriarchal thrones of the East, subjected to abbasid caliphate. They were directly involved by the emperor in his church-pacification project. This contribution analyzed the reactions of the two Seats that first sent their representatives to the council: Antioch and Jerusalem

René Girard e il ruolo degli oracoli nell'Edipo Re di Sofocle

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