Biomedical Engineering and Bioengineering Commons^™

Soil Respiration Measurements Reveal High Retention Of Organic Carbon From Corn Residue Derived High-Lignin Fermentation Byproduct Enabling Sustainable Lignocellulosic Biofuel Production, Michelle Sun Wang

Dartmouth College Master’s Theses

While 2G biofuel production can utilize non-edible, lignocellulosic feedstocks such as agricultural residues to produce liquid fuel, harvesting crop residues is unsustainable without careful management of the soil underneath. By harvesting a fraction of the crop residues left in the field after harvest, soil health can diminish and critically, the soil organic carbon (SOC) stored in agricultural fields can decrease. Currently, in the most popular 2G process models published, the issue of soil degradation remains unresolved with residue harvest strategies receiving considerable attention in the literature and other SOC management strategies receiving far less. Specifically, the strategy of returning the …

Ethanol-Induced Impacts On Nervous System Development And Dopamine Transporter Characterization In Schmidtea Mediterranea Planaria, Brennen Covely

Theses and Dissertations

Fetal alcohol spectrum disorders (FASD) are diagnosed in 2-5% of newborns, but the biology underlying FASD is poorly understood and challenging to study with existing model organisms. A new model for FASD research is Schmidtea mediterranea (Smed) planaria. Smed have a remarkable ability to regenerate their central nervous system (CNS), and possess a well-studied, simple genome. Previous studies have shown that ethanol exposure delays this regeneration, yet this relationship is not fully understood. Here, we show that alcohol exposure affects Smed in a dose-dependent manner, eliciting characteristic withdrawal-like behaviors and impacting cognitive function. Interestingly, prior exposure does not alter subsequent …

Design Of An Undergraduate Laboratory Experiment Utilizing A Stirred-Tank, Jacketed Bioreactor, Brennen Middleton

Honors Theses

Bioreactors are utilized in many industries, such as the food, alternative fuel, and pharmaceutical industries, to design and manufacture products. Unlike the similar chemical batch reactors utilized in many chemical processing facilities, bioreactor utilization requires engineers to consider a wider range of operating conditions and parameters. This results in a more complex reaction system and controls network. Due to this, it is imperative for chemical and biomedical engineering students to not only understand the theory surrounding these reactor systems, but also understand how to properly design and perform operating procedures with these systems. Thus, it was determined to create an …

Lipid Droplet Membrane Proteome Remodeling Parallels Ethanol-Induced Hepatic Steatosis And Its Resolution, Carol A. Casey, Terrence M. Donohue Jr., Jacy L. Kubik, Vikas Kumar, Michael J. Naldrett, Nicholas T. Woods, Cole P. Frisbie, Mark A. Mcniven, Paul G. Thomes

Nebraska Center for Biotechnology: Faculty and Staff Publications

Abstract Lipid droplets (LDs) are composed of neutral lipids enclosed in a phospholipid monolayer, which harbors membrane-associated proteins that regulate LD functions. Despite the crucial role of LDs in lipid metabolism, remodeling of LD protein composition in disease contexts, such as steatosis, remains poorly understood. We hypothesized that chronic ethanol consumption, subsequent abstinence from ethanol, or fasting differentially affects the LD membrane proteome content and that these changes influence how LDs interact with other intracellular organelles. Here, male Wistar rats were pair-fed liquid control or ethanol diets for 6 weeks, and then, randomly chosen animals from both groups were either …

High Gravity Fermentation Of Sugarcane Bagasse Hydrolysate By Saccharomyces Pastorianus To Produce Economically Distillable Ethanol Concentrations: Necessity Of Medium Components Examined, Sarah W. Harcum, Thomas P. Caldwell

Open Access Publishing Fund

A major economic obstacle in lignocellulosic ethanol production is the low sugar concentrations in the hydrolysate and subsequent fermentation to economically distillable ethanol concentrations. We have previously demonstrated a two-stage fermentation process that recycles xylose with xylose isomerase to increase ethanol productivity, where the low sugar concentrations in the hydrolysate limit the final ethanol concentrations. In this study, three approaches are combined to increase ethanol concentrations. First, the medium-additive requirements were investigated to reduce ethanol dilution. Second, methods to increase the sugar concentrations in the sugarcane bagasse hydrolysate were undertaken. Third, the two-stage fermentation process was recharacterized with high gravity …

Developing A Control System To Better Understand The Effects Of Pyruvate Decarboxylase Activity On Clostridium Thermocellum Metabolism, Nicholas Cervenka

ENGS 88 Honors Thesis (AB Students)

In order for cellulosic biofuels from Clostridium thermocellum to be commercially viable, the ethanol yield and titer of the microbe must be increased. To accomplish this, it has been suggested to introduce the Pyruvate Decarboxylase (PDC) enzyme into C. thermocellum. In order to demonstrate effects on ethanol production by PDC prior to genetic modification, a cell free system (CFS) has been developed. A purified enzyme system was developed with the CFS to function as a control. Using the purified enzyme system, PDC from Saccharomyces cerevisiae was demonstrated to be a good candidate for further testing in the CFS.

Storage Of Round And Square Switchgrass Bales: Effect Of Storage Time And Fungal Inoculation On Saccharification Efficiency, Noaa Frederick

Graduate Theses and Dissertations

To produce fermentable sugars from lignocellulosic biomass feedstock, severe pretreatment conditions are needed (either high acid concentration, temperature, or retention times). High severities can produce toxic byproducts which inhibit enzymatic hydrolysis or fermentation. In order to reduce pretreatment severities (and thus increase enzyme and fermentation efficiency), the white-rot fungus Pleurotus ostreastus was seeded into square and round bales of Kanlow switchgrass (Panicum virgastum L.) and left in the field over a period of 9 month. The laccase producing fungus is believed to selectively degrade lignin, a common plant structural polymer, which can function as an enzymatic inhibitor. Samples were taken …

Coordinated Dynamic Gene Expression Changes In The Central Nucleus Of The Amygdala During Alcohol Withdrawal., Kate Freeman, Mary M. Staehle, Rajanikanth Vadigepalli, Gregory E Gonye, Babatunde A Ogunnaike, Jan B Hoek, James S Schwaber

Faculty Scholarship for the College of Science & Mathematics

BACKGROUND: Chronic alcohol use causes widespread changes in the cellular biology of the amygdala's central nucleus (CeA), a GABAergic center that integrates autonomic physiology with the emotional aspects of motivation and learning. While alcohol-induced neurochemical changes play a role in dependence and drinking behavior, little is known about the CeA's dynamic changes during withdrawal, a period of emotional and physiologic disturbance.

METHODS: We used a qRT-PCR platform to measure 139 transcripts in 92 rat CeA samples from control (N = 33), chronically alcohol exposed (N = 26), and withdrawn rats (t = 4, 8, 18, 32, and 48 hours; N …

Mutant Alcohol Dehydrogenase Leads To Improved Ethanol Tolerance In Clostridium Thermocellum, Steven D. Brown, Adam M. Guss, Tatiana V. Karpinets, Jerry M. Parks

Dartmouth Scholarship

Clostridium thermocellum is a thermophilic, obligately anaerobic, Gram-positive bacterium that is a candidate microorganism for converting cellulosic biomass into ethanol through consolidated bioprocessing. Ethanol intolerance is an important metric in terms of process economics, and tolerance has often been described as a complex and likely multigenic trait for which complex gene interactions come into play. Here, we resequence the genome of an ethanol-tolerant mutant, show that the tolerant phenotype is primarily due to a mutated bifunctional acetaldehyde-CoA/alcohol dehydrogenase gene (adhE), hypothesize based on structural analysis that cofactor specificity may be affected, and confirm this hypothesis using enzyme assays. …

Life Cycle Assessment Of Greenhouse Gas Emissions From Ethanol And Biopolymers, Adam J. Liska, Xiao Xue Fang

Adam Liska Papers

Conclusions

• Regulatory LCA is not likely to be used for non‐fuel chemicals alone in the near future

• Significant GHG emission credits for corn‐ethanol can be obtained by using only roughly 6‐9% of initial starch for production of biopolymers based on previous LCA theory

• Pay close attention to values in calculating credits per kg—these have to stand up in litigation to ensure the credit

• Credits are proportional to the mass of polymer produced

• Many theoretical issues are uncertain and credits will only be determined in conjunction with EPA

• Indirect emissions are uncertain and are a …

Ethanol Yields And Cell Wall Properties In Divergently Bred Switchgrass Genotypes, Gautam Sarath, Bruce S. Dien, Aaron J. Saathoff, Kenneth P. Vogel, Robert B. Mitchell, Han Chen

Biological Systems Engineering: Papers and Publications

Genetic modification of herbaceous plant cell walls to increase biofuels yields is a primary bioenergy research goal. Using two switchgrass populations developed by divergent breeding for ruminant digestibility, the contributions of several wall-related factors to ethanol yields was evaluated. Field grown low lignin plants significantly out yielded high lignin plants for conversion to ethanol by 39.1% and extraction of xylans by 12%. However, across all plants analyzed, greater than 50% of the variation in ethanol yields was attributable to changes in tissue and cell wall architecture, and responses of stem biomass to diluteacid pretreatment. Although lignin levels were lower in …

Evaluation Of Ethanol And Water Introduction Via Fumigation On Efficiency And Emissions Of A Compression Ignition Engine Using An Atomization Technique, Grant S. Janousek

Department of Agricultural and Biological Systems Engineering: Dissertations, Theses, and Student Research

Performance of a diesel engine, equipped for ethanol and water fumigation, was studied. The method implemented allowed for non-destructive introduction of liquids in advance of the turbocharger. Engine torque, speed, emission components, diesel and ethanol fuel rates were recorded and analyzed for each mixture of inputs. Based on the results of the study, thermal efficiency was not significantly different from the baseline diesel performance when using several ethanol and water mixtures. On the other hand, ethanol fumigation caused a significant reduction in NOx emissions and an increase in HC and CO emissions. No significant changes in CO2 or O2 occurred.

Integration Of Microfluidics With Surface Plasmon Resonance, Scott B. Fratzke

Master's Theses

This thesis successfully integrates laminate microfluidic devices with an analytic Surface Plasmon Resonance (SPR) instrument. Integration was accomplished at low-cost using materials such as polydimethylsiloxane (PDMS), Poly(methyl methacrylate) (PMMA), Tygon tubing, and a 3-way stopcock. The main components of this thesis are the design and fabrication of the low-cost, in-house fluidics that can integrate with upstream microfluidics and the validation of the in-house fluidics using the Biosensing Instruments BI-2000 SPR instrument. The low-cost fluidics was designed and fabricated “in-house” using a novel investment casting technique that required the use of laser cutting technology to make a master cast, and candle …

Switchgrass For Bioethanol And Other Value-Added Applications: A Review, Deepak R. Keshwani, Jay J. Cheng

Biological Systems Engineering: Papers and Publications

Switchgrass is a promising feedstock for value-added applications due to its high productivity, potentially low requirements for agricultural inputs and positive environmental impacts. The objective of this paper is to review published research on the conversion of switchgrass into bioethanol and other value-added products. Environmental benefits associated with switchgrass include the potential for carbon sequestration, nutrient recovery from runoff, soil remediation and provision of habitats for grassland birds. Pretreatment of switchgrass is required to improve the yields of fermentable sugars. Based on the type of pretreatment, glucose yields range from 70% to 90%, and xylose yields range from 70% to …

Antiproliferation Properties Of Grain Sorghum Dry Distiller’S Grain Lipids In Caco-2 Cells, Richard Zbasnik, Timothy P. Carr, Curtis L. Weller, Keum Taek Hwang, Lijun Wang, Susan L. Cuppett, Vicki Schlegel

Biological Systems Engineering: Papers and Publications

Antiproliferative properties of lipids extracted from grain sorghum (GS) dry distiller’s grain (DDG) were analyzed to determine the feasibility of developing GS coproducts as a source for human health dietary ingredients. The lipid extract of GSDDG was delivered to human colon carcinoma (Caco-2) cells by solubilizing 0−1000 μg/mL of GS-DDG lipids in 100 μg/mL increments with micelles. A significant reduction in cell viability (25−50%) resulted at treatment levels of 400−1000 μg/mL GS-DDG lipids (p < 0.05). Alternatively, total protein levels of cells treated with 400, 500, and 600 μg/mL of GS-DDG lipid were not significantly different from the control, indicating cell growth during the treatment period. Total cell counts for the control were not significantly different from the GS-DDG lipid treated cells, but dead cell counts increased by 10% for the latter sample with a concomitant increase of the intercellular protein lactate dehydrogenase leakage (30−40%) in the medium. Preliminary analysis by the fluorescence-activated cell method (FACs) demonstrated that nonviable cells were in either the early apoptotic, late apoptotic, or necrotic stage post-treatment with 400, 500, and 600 μg/mL GS-DDG lipids. Physiochemical characterization of the GS-DDG lipids used for the antiproliferation study showed the presence of vitamin E (predominantly γ-tocopherol), triacylglycerides (predominantly linoleic acid), policosanols, aldehydes, and sterols (predominantly campesterol and stigmasterol), each of which or as synergistic/additive group of constituents may be responsible for the antiproliferative effect.

Metabolic Engineering Of A Thermophilic Bacterium To Produce Ethanol At High Yield, A. Joe Shaw, Kara K. Podkaminer, Sunil G. Desai, John S. Bardsley, Stephen R. Rogers, Philip G. Thorne, David A. Hogsett, Lee R. Lynd

Dartmouth Scholarship

We report engineering Thermoanaerobacterium saccharolyticum, a thermophilic anaerobic bacterium that ferments xylan and biomass-derived sugars, to produce ethanol at high yield. Knockout of genes involved in organic acid formation (acetate kinase, phosphate acetyltransferase, and L-lactate dehydrogenase) resulted in a strain able to produce ethanol as the only detectable organic product and substantial changes in electron flow relative to the wild type. Ethanol formation in the engineered strain (ALK2) utilizes pyruvate:ferredoxin oxidoreductase with electrons transferred from ferredoxin to NAD(P), a pathway different from that in previously described microbes with a homoethanol fermentation. The homoethanologenic phenotype was stable for >150 generations …

Adenosine Type 1 (A ) Receptors Mediate Protection Against Myocardial 1 Infarction Produced By Chronic, Intermittent Ingestion Of Ethanol In Dogs, Franz Kehl, John G. Krolikowski, John F. Ladisa, Judy R. Kersten, David C. Warltier, Paul S. Pagel

Biomedical Engineering Faculty Research and Publications

Background: Chronic consumption of small amounts of ethanol protects myocardium from ischemic injury. We tested the hypothesis that adenosine type 1 (A₁) receptors mediate these beneficial effects.

Methods: Dogs (n=37) were fed with ethanol (1.5 g/kg) or water mixed with dry food twice per day for 12 weeks, fasted overnight before experimentation, and instrumented for measurement of hemodynamics. Dogs received intravenous drug vehicle (50% polyethylene glycol in 0.1 N sodium hydroxide and 0.9% saline over 15 min) or the selective A₁ receptor antagonist 8-cyclopentyl-1,3-dipropylxanthine (DPCPX, 0.8 mg/kg over 15 min) and were subjected …