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Investigating the Role of the Las and Rhl Quorum Sensing Systems in the Pathogenesis of Pseudomonas aeruginosa

Description

Pseudomonas aeruginosa is an opportunistic bacterial pathogen commonly associated with increased morbidity and mortality in cystic fibrosis (CF) patients. To adapt to the CF lung environment, P. aeruginosa undergoes multiple genetic changes as it moves from an acute to a

Pseudomonas aeruginosa is an opportunistic bacterial pathogen commonly associated with increased morbidity and mortality in cystic fibrosis (CF) patients. To adapt to the CF lung environment, P. aeruginosa undergoes multiple genetic changes as it moves from an acute to a chronic infection. The resultant phenotypes have been associated with chronic infection and can provide important information to track the patient’s individualized disease progression. This study examines the link between the accumulation of QS genetic mutations and phenotypic expression in P. aeruginosa laboratory strains and clinical isolates. We utilized several plate-based and colorimetric assays to quantify the production of pyocyanin, rhamnolipids, and protease from paired clinical early- and late-stage chronic infection isolates across 16 patients. Exoproduct production of each isolate was compared to the mean production of pooled isolates to classify high producing (QS-sufficient) and low producing (QS-deficient) isolates. We found that over time P. aeruginosa isolates exhibit a reduction in QS-related phenotypes during chronic infections. Future research of the QS regulatory networks will identify whether reversion of genotype will result in corresponding phenotypic changes in QS-deficient chronic infection isolates.

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2020-05

A Reflection on Translating a Thesis on Soil Pollution in Jiangsu

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This project took thesis written in Mandarin researching heavy metal pollution in the Jiangsu region of province and translated it to English. Then the reflection process was discussed, considering the translation challenges between Mandarin and English and how the scientific nature of the piece played into that process.

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2020-12

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Computational Modeling and Experimentation of Pervaporation Membrane Processes for Brackish Water Recovery

Description

Fresh water is essential to the human population and is an integral component in global economics for its multiple uses, and population growth/development cause concern for the possible exhaustion of the limited supply of freshwater. A combined computational and experimental

Fresh water is essential to the human population and is an integral component in global economics for its multiple uses, and population growth/development cause concern for the possible exhaustion of the limited supply of freshwater. A combined computational and experimental approach to observe and evaluate pervaporation membrane performance for brackish water recovery was done to assess its efficiency and practicality for real world application. Results from modeling conveyed accuracy to reported parameter values from literature as well as strong dependence of performance on input parameters such as temperature. Experimentation results showed improved performance in flux by 34%-42% with radiative effect and then additional performance improvement (9%-33%) with the photothermal effect from carbon black application. Future work will include improvements to the model to include scaling propensity and energy consumption as well as continued experimentation to assess quality of pervaporation in water recovery.

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2018-05

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The History, Evolution, and Development of Wedding Cakes

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A key component to American weddings is the wedding cake and the number of customs associated with it. The History, Evolution, and Development of Wedding Cakes is a creative project that hopes to answer why the wedding cake is such

A key component to American weddings is the wedding cake and the number of customs associated with it. The History, Evolution, and Development of Wedding Cakes is a creative project that hopes to answer why the wedding cake is such an iconic item and what it represents. A historical study details the evolution from its origin to present day while a comparison of wedding cakes (or wedding cake analogs) from other cultures explores their role(s) across the globe. It was found that while little regard is given to the symbolism of the wedding cake today, its presence persists in some shape or form as it continues to evolve. Cultural variations were recreated to reflect personal interpretations of contemporary designs in addition to an original design of a contemporary-rustic American wedding cake.

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2018-05

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Identification of aromatic-inducible promoters and heterologous biosensors as tuning elements for styrene production in E. coli

Description

One of the primary bottlenecks to chemical production in biological organisms is the toxicity of the chemical. Overexpression of efflux pumps has been shown to increase tolerance to aromatic compounds such as styrene and styrene oxide. Tight control of pum

One of the primary bottlenecks to chemical production in biological organisms is the toxicity of the chemical. Overexpression of efflux pumps has been shown to increase tolerance to aromatic compounds such as styrene and styrene oxide. Tight control of pump expression is necessary to maximize titers and prevent excessive strain on the cells. This study aimed to identify aromatic-sensitive native promoters and heterologous biosensors for construction of closed-loop control of efflux pump expression in E. coli. Using a promoter library constructed by Zaslaver et al., activation was measured through GFP output. Promoters were evaluated for their sensitivity to the addition of one of four aromatic compounds, their "leaking" of signal, and their induction threshold. Out of 43 targeted promoters, 4 promoters (cmr, mdtG, yahN, yajR) for styrene oxide, 2 promoters (mdtG, yahN) for styrene, 0 promoters for 2-phenylethanol, and 1 promoter for phenol (pheP) were identified as ideal control elements in aromatic bioproduction. In addition, a series of three biosensors (NahR, XylS, DmpR) known to be inducible by other aromatics were screened against styrene oxide, 2-phenylethanol, and phenol. The targeted application of these biosensors is aromatic-induced activation of linked efflux pumps. All three biosensors responded strongly in the presence of styrene oxide and 2-phenylethanol, with minor activation in the presence of phenol. Bioproduction of aromatics continues to gain traction in the biotechnology industry, and the continued discovery of aromatic-inducible elements will be essential to effective pathway control.

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2018-05

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Mass Transfer Kinetics of Novel Asymmetric Hollow-fiber Membranes

Description

This report investigates the mass-transfer kinetics of gas diffusion through an asymmetrical hollow-fiber membrane developed for the membrane biofilm reactor (MBfR) when it is used to microbiologically convert syngas (a mixture of H2, CO2, and CO) to organic products.

This report investigates the mass-transfer kinetics of gas diffusion through an asymmetrical hollow-fiber membrane developed for the membrane biofilm reactor (MBfR) when it is used to microbiologically convert syngas (a mixture of H2, CO2, and CO) to organic products. The asymmetric Matrimid® membrane had superior diffusion fluxes compared to commercially available symmetric, three-layer composite and polypropylene single-layer membranes. The Matrimid® asymmetric membrane had a H2 gas-gas diffusion flux between 960- and 1600-fold greater than that of the composite membrane and between 32,000- and 46,800-fold greater than that of the single-layer membrane. Gas-gas diffusion experiments across the Matrimid® membrane also demonstrated plasticization behavior for pure CO2 and H2 gas feeds. In particular, a 10 psia increase in inlet pressure resulted in a 12-fold increase in permeance for H2 and a 16-fold increase for CO2. Plasticization was minimal for symmetric composite and single-layer membranes. Thus, diffusion fluxes were much higher for the asymmetric membrane than for the symmetric composite and single-layer membranes, and this supports the promise of the asymmetric membrane as a high-efficiency means to deliver syngas to biofilms able to convert the syngas to organic products. Gas-liquid diffusion was much slower than gas-gas diffusion, and this supports the benefit of using the MBfR approach over fermentation reactors that rely on sparging syngas.

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2018-05

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New Diagnostic Methods for Detecting Microvillus Inclusion Disease

Description

Microvillus Inclusion disease is a fatal disease found in the Navajo population caused by a single nucleotide polymorphism. It is characterized by intractable diarrhea and is often fatal early in life.1 The current method of diagnosis is sending duodenal biopsies

Microvillus Inclusion disease is a fatal disease found in the Navajo population caused by a single nucleotide polymorphism. It is characterized by intractable diarrhea and is often fatal early in life.1 The current method of diagnosis is sending duodenal biopsies for histopathological examination and confirmatory testing through genomic sequencing. The purpose of this experiment was to create a more simple and cost-effective diagnostic method for detecting Microvillus Inclusion disease. Three methods were explored (RFLP2, ARMS3,4, and Tentacle Probes5,6) and two methods were tested to determine their ability and their efficiency in detecting the SNP that causes the disease.2 Tests using the RFLP2 method and synthetic DNA resulted in 9% false-positive rate and 11% false-negative rate in a blind trial for detecting both target (mutation present) and non-target (mutation absent) DNA when gel analyzing software was used to compare Rf values after gel electrophoresis. Using the ARMS method3, a nine-sample randomized test was run that ended up with 22% false-positive rate and 19% false-negative rate from a blind trial when using a gel analyzing software to determine presence of the SNP by band intensity. Disclaimer: No DNA from human patients was used in this study. Only synthetic DNA used.

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2018-05

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Methods to Enhance Overexpression of Membrane Protein in Escherichia coli

Description

Membrane proteins (MPs) are an important aspect of cell survival that ensure structural integrity, signaling, and transportation of molecules. Since 2015, over 450 MPs have been studied to find their functionalities and structure. Sufficient amounts of correctly folded MPs are

Membrane proteins (MPs) are an important aspect of cell survival that ensure structural integrity, signaling, and transportation of molecules. Since 2015, over 450 MPs have been studied to find their functionalities and structure. Sufficient amounts of correctly folded MPs are needed to accurately study them through crystallography and other structural study methods. Use of recombinant technology is needed to overexpress MPs as natural abundance of MP is often too slow to provide the necessary amounts. However, an increase in toxicity and decrease in generation time deter the overexpression of MPs. The following report discusses two methods of enhancing overexpression in Escherchia coli, the use of T7 RNA polymerase (T7RNAP) and the reprogramming of chaperon pathways, that combats toxicity and promotes cell growth. Overall, both methods are proven to work effectively to overexpress MPs by regulating transcription rate of mRNA (T7RNAP) or folding and transporting of polypeptides to inner membrane (chaperon pathway). To further study the effectiveness of the two methods, they will need to be compared at the same conditions. In addition, a combination of two methods should also be studied to find out if the combination would have a great impact on the overexpression of the MPs.

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2018-05

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Conductive Heat Transfer in Rotary Drums

Description

In industrial applications, rotary drums are poorly understood and preform suboptimally when used to process particulates. In order to better understand how these drums work, a statistical experiment was designed to measure the effects of the fill level and rotation

In industrial applications, rotary drums are poorly understood and preform suboptimally when used to process particulates. In order to better understand how these drums work, a statistical experiment was designed to measure the effects of the fill level and rotation rate on the final temperature of the particle bed. A steel rotary drum was set up to be headed by three external heat guns, simulating the conditions under which standard rotary drums are operated. By measuring the bed temperature at steady state, and recording the combination of factors in each run, a regression analysis was run to determine the factor's effects. Fill level was seen to have a small positive effect, rotation rate was seen to have a small negative effect, and the interaction of the two was shown to have a large positive effect. This led the team to conclude that the flow profile of the bed may be the most important factor in heat transfer, and that further research should be done to isolate and study the effect of the flow profile.

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2018-05

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An Examination of the Impact of Support Design on 316 Stainless Steel Supports

Description

The removal of support material from metal 3D printed objects is a laborious necessity for the post-processing of powder bed fusion printing (PBF). Supports are typically mechanically removed by machining techniques. Sacrificial supports are necessary in PBF printing to relieve

The removal of support material from metal 3D printed objects is a laborious necessity for the post-processing of powder bed fusion printing (PBF). Supports are typically mechanically removed by machining techniques. Sacrificial supports are necessary in PBF printing to relieve thermal stresses and support overhanging parts often resulting in the inclusion of supports in regions of the part that are not easily accessed by mechanical removal methods. Recent innovations in PBF support removal include dissolvable metal supports through an electrochemical etching process. Dissolvable PBF supports have the potential to significantly reduce the costs and time associated with traditional support removal. However, the speed and effectiveness of this approach is inhibited by numerous factors such as support geometry and metal powder entrapment within supports. To fully realize this innovative approach, it is necessary to model and understand the design parameters necessary to optimize support structures applicable to an electrochemical etching process. The objective of this study was to evaluate the impact of block additive manufacturing support parameters on key process outcomes of the dissolution of 316 stainless steel support structures. The parameters investigated included hatch spacing and perforation, and the outcomes of interests included time required for completion, surface roughness, and effectiveness of the etching process. Electrical current was also evaluated as an indicator of process completion. Analysis of the electrical current throughout the etching process showed that the dissolution is diffusion limited to varying degrees, and is dependent on support structure parameters. Activation and passivation behavior was observed during current leveling, and appeared to be more pronounced in non-perforated samples with less dense hatch spacing. The correlation between electrical current and completion of the etching process was unclear, as the support structures became mechanically removable well before the current leveled. The etching process was shown to improve surface finish on unsupported surfaces, but support was shown to negatively impact surface finish. Tighter hatch spacing was shown to correlate to larger variation in surface finish, due to ridges left behind by the support structures. In future studies, it is recommended current be more closely correlated to process completion and more roughness data be collected to identify a trend between hatch spacing and surface roughness.

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2018-05