Comparative Efficiency and Driving Range of Light- and Heavy-Duty Vehicles Powered with Biomass Energy Stored in Liquid Fuels or Batteries

This study addresses the question, When using cellulosic biomass for vehicular transportation, which field-to-wheels pathway is more efficient: that using biofuels or that using bioelectricity? In considering the question, the level of assumed technological maturity significantly affects the comparison, as does the intended transportation application. Results from the analysis indicate that for light-duty vehicles, over ranges typical in the United States today (e.g., 560-820 miles), field-to-wheels performance is similar, with some scenarios showing biofuel to be more efficient, and others indicating the two pathways to be essentially the same. Over the current range of heavy-duty vehicles, the field-to-wheels efficiency is higher for biofuels than for electrically powered vehicles. Accounting for technological advances and range, there is little basis to expect mature bioelectricity-powered vehicles to have greater field-to-wheels efficiency (e.g., kilometers per gigajoule biomass or per hectare) compared with mature biofuel-powered vehicles

Acceptable Risk

Perhaps the topic of acceptable risk never had a sexier and more succinct introduction than the one Edward Norton, playing an automobile company executive, gave it in Fight Club: “Take the number of vehicles in the field (A), multiply it by the probable rate of failure (B), and multiply the result by the average out of court settlement (C). A*B*C=X. If X is less than the cost of the recall, we don’t do one.” Of course, this dystopic scene also gets to the heart of the issue in another way: acceptable risk deals with mathematical calculations about the value of life, injury, and emotional wreckage, making calculation a difficult matter ethically, politically, and economically. This entry will explore the history of this idea, focusing on its development alongside statistics into its wide importance today

Computational Design and Characterization of a Temperature-Sensitive Plasmid Replicon for Gram Positive Thermophiles

Temperature-sensitive (Ts) plasmids are useful tools for genetic engineering, but there are currently none compatible with the gram positive, thermophilic, obligate anaerobe, Clostridium thermocellum. Traditional mutagenesis techniques yield Ts mutants at a low frequency, and therefore requires the development of high-throughput screening protocols, which are also not available for this organism. Recently there has been progress in the development of computer algorithms which can predict Ts mutations. Most plasmids currently used for genetic modification of C. thermocellum are based on the replicon of plasmid pNW33N, which replicates using the RepB replication protein. To address this problem, we set out to create a Ts plasmid by mutating the gene coding for the RepB replication protein using an algorithm designed by Varadarajan et al. (1996) for predicting Ts mutants based on the amino-acid sequence of the protein

Kinetics and Relative Importance of Phosphorolytic and Hydrolytic Cleavage of Cellodextrins and Cellobiose in Cell Extracts of Clostridium thermocellum

Rates of phosphorolytic cleavage of -glucan substrates were determined for cell extracts from Clostridium thermocellum ATCC 27405 and were compared to rates of hydrolytic cleavage. Reactions with cellopentaose and cellobiose were evaluated for both cellulose (Avicel)- and cellobiose-grown cultures, with more limited data also obtained for cellotetraose. To measure the reaction rate in the chain-shortening direction at elevated temper- atures, an assay protocol was developed featuring discrete sampling at 60°C followed by subsequent analysis of reaction products (glucose and glucose-1-phosphate) at 35°C. Calculated rates of phosphorolytic cleavage for cell extract from Avicel-grown cells exceeded rates of hydrolytic cleavage by \u3e20-fold for both cellobiose and cellopentaose over a 10-fold range of -glucan concentrations (0.5 to 5 mM) and for cellotetraose at a single concentration (2 mM). Rates of phosphorolytic cleavage of -glucosidic bonds measured in cell extracts were similar to rates observed in growing cultures. Comparisons of Vmax values indicated that cellobiose- and cellodextrin-phosphorylating activities are synthesized during growth on both cellobiose and Avicel but are subject to some degree of metabolic control. The apparent Km for phosphorolytic cleavage was lower for cellopentaose (mean value for Avicel- and cellobiose-grown cells, 0.61 mM) than for cellobiose (mean value, 3.3 mM)

Accept me for who I am! : a critical ethnographic study of a participatory research project with people labeled mentally retarded.

This dissertation is a critical ethnographic study of a participatory research project in which a group of eight adults labeled mentally retarded, with the assistance of two nondisabled adults, created and performed a musical theater production called Special. Special was produced as part of a participatory research process in which group members also interviewed friends, advocates of disabled people, and former residents of a local institution for people labeled retarded, in order to find out how ex-residents were treated once they were placed in community living situations. The information from these interviews, as well as accounts from group members\u27 own lives, comprised the content of Special. This study consists of two main parts--an interpretive section (Chapters 5 and 6), including emic and etic interpretations of group members\u27 experiences, and a critical section (Chapter 7), in which an internalized oppression framework is invoked to examine group members\u27 experiences. Three main findings of the study were: (1) that group members expressed a chronic problem orientation; (2) that group members exhibited a justice orientation; and (3) that group members were largely motivated by the drive to visibility, or the need to be seen, understood and accepted for who they really are. Another major finding of the study was that group members\u27 drive to visibility was not only a major motivation for doing the play, but was also a key to understanding much of their behavior--that when they felt visible, they acted up, or became positive and productive, and that when they felt invisible, they acted out, or became destructive, and even violent, evidence of internalized oppression in group members. Group members\u27 drive to visibility, coupled with their resistance to an identity of mental impairment, raises two important questions regarding the issue of social identity with people labeled retarded: (1) Are there reasons to believe that people labeled retarded can feel a sense of pride in who they are, both as individuals and as members of a social group? (2) If people labeled retarded cannot feel a sense of pride, what are their prospects of overcoming internalized oppression, and of working with one another as a group with an identity, a purpose, and a right to have power like all other groups

Identification of the [FeFe]-Hydrogenase Responsible for Hydrogen Generation in Thermoanaerobacterium saccharolyticum and Demonstration of Increased Ethanol Yield via Hydrogenase Knockout

Three putative hydrogenase enzyme systems in Thermoanaerobacterium saccharolyticum were investigated at the genetic, mRNA, enzymatic, and phenotypic levels. A four-gene operon containing two [FeFe]-hydrogenase genes, provisionally termed hfs (hydrogenase-Fe-S), was found to be the main enzymatic catalyst of hydrogen production. hfsB, perhaps the most interesting gene of the operon, contains an [FeFe]-hydrogenase and a PAS sensory domain and has several conserved homologues among clostridial saccharolytic, cellulolytic, and pathogenic bacteria. A second hydrogenase gene cluster, hyd, exhibited methyl viologen-linked hydrogenase enzymatic activity, but hyd gene knockouts did not influence the hydrogen yield of cultures grown in closed-system batch fermentations. This result, combined with the observation that hydB contains NAD(P)+ and FMN binding sites, suggests that the hyd genes are specific to the transfer of electrons from NAD(P)H to hydrogen ions. A third gene cluster, a putative [NiFe]-hydrogenase with homology to the ech genes, did not exhibit hydrogenase activity under any of the conditions tested. Deletion of the hfs and hydA genes resulted in a loss of detectable methyl viologen-linked hydrogenase activity. Strains with a deletion of the hfs genes exhibited a 95% reduction in hydrogen and acetic acid production. A strain with hfs and ldh deletions exhibited an increased ethanol yield from consumed carbohydrates and represents a new strategy for engineering increased ethanol yields in T. saccharolyticum

Natural Competence in Thermoanaerobacter and Thermoanaerobacterium Species

Low-G+C thermophilic obligate anaerobes in the class Clostridia are considered among the bacteria most resistant to genetic engineering due to the difficulty of introducing foreign DNA, thus limiting the ability to study and exploit their native hydrolytic and fermentative capabilities. Here, we report evidence of natural genetic competence in 13 Thermoanaerobacter and Thermoanaerobacterium strains previously believed to be difficult to transform or genetically recalcitrant. In Thermoanaerobacterium saccharolyticum JW/SL-YS485, natural competence- mediated DNA incorporation occurs during the exponential growth phase with both replicating plasmid and homologous recombination-based integration, and circular or linear DNA. In T. saccharolyticum, disruptions of genes similar to comEA, comEC, and a type IV pilus (T4P) gene operon result in strains unable to incorporate further DNA, suggesting that natural competence occurs via a conserved Gram-positive mechanism. The relative ease of employing natural competence for gene transfer should foster genetic engineering in these industrially relevant organisms, and understanding the mechanisms underlying natural competence may be useful in increasing the applicability of genetic tools to difficult-to-transform organisms

Formation and Characterization of Non-Growth States in Clostridium Thermocellum: Spores and L-Forms

Clostridium thermocellum is an anaerobic thermophilic bacterium that exhibits high levels of cellulose solublization and produces ethanol as an end product of its metabolism. Using cellulosic biomass as a feedstock for fuel production is an attractive prospect, however, growth arrest can negatively impact ethanol production by fermentative microorganisms such as C. thermocellum. Understanding conditions that lead to non-growth states in C. thermocellum can positively influence process design and culturing conditions in order to optimize ethanol production in an industrial setting