Functionalized carbon nanotube nanoelectrodes for biomolecular recognition

The objective of this research is to develop nanoscale ultrasensitive transducers for detection of biological species at molecular level using carbon nanotubes as nanoelectrodes. Rapid detection of ultra low concentration or even single DNA molecules are essential for medical diagnosis and treatment, pharmaceutical applications, gene sequencing as well as forensic analysis. Here the use of functionalized single walled carbon nanotubes (SWNT) as nanoscale detection platform for rapid detection of single DNA molecules is demonstrated. The detection principle is based on obtaining electrical signal from a single amine terminated DNA molecule which is covalently bridged between two ends of an SWNT separated by a nanoscale gap. The synthesis, fabrication, chemical functionalization of nanoelectrodes and DNA attachment were optimized to perform reliable electrical characterization these molecules. Using this detection system fundamental study on charge transport in DNA molecule of both genomic and non genomic sequences is performed. We measured an electrical signal of about 30 pA through a hybridized DNA molecule of 80 base pair in length which encodes a portion of sequence of H5N1 gene of avian Influenza A virus. Due the dynamic nature of the DNA molecules the local environment such as ion concentration, pH and temperature significantly influence its physical properties. We observed a decrease in DNA conductance of about 33% in high vacuum conditions. The counterion variation was analyzed by changing the buffer from sodium acetate to tris(hydroxymethyl) aminomethane, which resulted in a two orders of magnitude increase in the conductivity of the DNA. The fabrication of large array of identical SWNT nanoelectrodes was achieved by using ultralong SWNTs. Using these nanoelectrode array we have investigated the sequence dependent charge transport in DNA. A systematic study performed on PolyG - PolyC sequence with varying number of intervening PolyA - PolyT pairs showed a decrease in electrical signal from 180 pA (PolyG - PolyC) to 30 pA with increasing number of the PolyA - PolyT pairs. This work also led to the development of ultrasensitive nanoelectrodes based on enzyme functionalized vertically aligned high density multiwalled CNTs for electrochemical detection of cholesterol. The nanoelectrodes exhibited selectively detection of cholesterol in the presence of common interferents found in human blood.

Assessment of the occurrence and potential risks of antibiotics and their metabolites in South Florida waters using liquid chromatography tandem mass spectrometry

An automated on-line SPE-LC-MS/MS method was developed for the quantitation of multiple classes of antibiotics in environmental waters. High sensitivity in the low ng/L range was accomplished by using large volume injections with 10-mL of sample. Positive confirmation of analytes was achieved using two selected reaction monitoring (SRM) transitions per antibiotic and quantitation was performed using an internal standard approach. Samples were extracted using online solid phase extraction, then using column switching technique; extracted samples were immediately passed through liquid chromatography and analyzed by tandem mass spectrometry. The total run time per each sample was 20 min. The statistically calculated method detection limits for various environmental samples were between 1.2 and 63 ng/L. Furthermore, the method was validated in terms of precision, accuracy and linearity. ^ The developed analytical methodology was used to measure the occurrence of antibiotics in reclaimed waters (n=56), surface waters (n=53), ground waters (n=8) and drinking waters (n=54) collected from different parts of South Florida. In reclaimed waters, the most frequently detected antibiotics were nalidixic acid, erythromycin, clarithromycin, azithromycin trimethoprim, sulfamethoxazole and ofloxacin (19.3-604.9 ng/L). Detection of antibiotics in reclaimed waters indicates that they can't be completely removed by conventional wastewater treatment process. Furthermore, the average mass loads of antibiotics released into the local environment through reclaimed water were estimated as 0.248 Kg/day. Among the surface waters samples, Miami River (reaching up to 580 ng/L) and Black Creek canal (up to 124 ng/L) showed highest concentrations of antibiotics. No traces of antibiotics were found in ground waters. On the other hand, erythromycin (monitored as anhydro erythromycin) was detected in 82% of the drinking water samples (n.d-66 ng/L). The developed approach is suitable for both research and monitoring applications.^ Major metabolites of antibiotics in reclaimed wates were identified and quantified using high resolution benchtop Q-Exactive orbitrap mass spectrometer. A phase I metabolite of erythromycin was tentatively identified in full scan based on accurate mass measurement. Using extracted ion chromatogram (XIC), high resolution data-dependent MS/MS spectra and metabolic profiling software the metabolite was identified as desmethyl anhydro erythromycin with molecular formula C36H63NO12 and m/z 702.4423. The molar concentration of the metabolite to erythromycin was in the order of 13 %. To my knowledge, this is the first known report on this metabolite in reclaimed water. Another compound acetyl-sulfamethoxazole, a phase II metabolite of sulfamethoxazole was also identified in reclaimed water and mole fraction of the metabolite represent 36 %, of the cumulative sulfamethoxazole concentration. The results were illustrating the importance to include metabolites also in the routine analysis to obtain a mass balance for better understanding of the occurrence, fate and distribution of antibiotics in the environment. ^ Finally, all the antibiotics detected in reclaimed and surface waters were investigated to assess the potential risk to the aquatic organisms. The surface water antibiotic concentrations that represented the real time exposure conditions revealed that the macrolide antibiotics, erythromycin, clarithromycin and tylosin along with quinolone antibiotic, ciprofloxacin were suspected to induce high toxicity to aquatic biota. Preliminary results showing that, among the antibiotic groups tested, macrolides posed the highest ecological threat, and therefore, they may need to be further evaluated with, long-term exposure studies considering bioaccumulation factors and more number of species selected. Overall, the occurrence of antibiotics in aquatic environment is posing an ecological health concern.^

Fiber Scaffolds of Poly (glycerol-dodecanedioate) and its Derivative via Electrospinning for Neural Tissue Engineering

Peripheral nerves have demonstrated the ability to bridge gaps of up to 6 mm. Peripheral Nerve System injury sites beyond this range need autograft or allograft surgery. Central Nerve System cells do not allow spontaneous regeneration due to the intrinsic environmental inhibition. Although stem cell therapy seems to be a promising approach towards nerve repair, it is essential to use the distinct three-dimensional architecture of a cell scaffold with proper biomolecule embedding in order to ensure that the local environment can be controlled well enough for growth and survival. Many approaches have been developed for the fabrication of 3D scaffolds, and more recently, fiber-based scaffolds produced via the electrospinning have been garnering increasing interest, as it offers the opportunity for control over fiber composition, as well as fiber mesh porosity using a relatively simple experimental setup. All these attributes make electrospun fibers a new class of promising scaffolds for neural tissue engineering. Therefore, the purpose of this doctoral study is to investigate the use of the novel material PGD and its derivative PGDF for obtaining fiber scaffolds using the electrospinning. The performance of these scaffolds, combined with neural lineage cells derived from ESCs, was evaluated by the dissolvability test, Raman spectroscopy, cell viability assay, real time PCR, Immunocytochemistry, extracellular electrophysiology, etc. The newly designed collector makes it possible to easily obtain fibers with adequate length and integrity. The utilization of a solvent like ethanol and water for electrospinning of fibrous scaffolds provides a potentially less toxic and more biocompatible fabrication method. Cell viability testing demonstrated that the addition of gelatin leads to significant improvement of cell proliferation on the scaffolds. Both real time PCR and Immunocytochemistry analysis indicated that motor neuron differentiation was achieved through the high motor neuron gene expression using the metabolites approach. The addition of Fumaric acid into fiber scaffolds further promoted the differentiation. Based on the results, this newly fabricated electrospun fiber scaffold, combined with neural lineage cells, provides a potential alternate strategy for nerve injury repair.

A Cultural Historical Study of Land Use in Dade County, Florida

Humans have used the land within the area currently defined as Dade County, Florida since around 11,000 B.P., but did not significantly alter the local environment until less than one hundred years ago. These recent changes greatly affected many critical ecological factors, thereby reducing the sustainability of many types of life, including humans. This study explains how land use evolved from earlier sustainable systems for satisfying human needs into the current menacing patterns. This is done by examining the environmental, technological, social, and cognitive contexts of land use through time. Changes in all these areas have followed a general trend leading to increasing intensity of land use and environmental change driven by population growth and technological innovation.

Fiber scaffolds of poly (glycerol-dodecanedioate) and its derivative via electrospinning for neural tissue engineering

Peripheral nerves have demonstrated the ability to bridge gaps of up to 6 mm. Peripheral Nerve System injury sites beyond this range need autograft or allograft surgery. Central Nerve System cells do not allow spontaneous regeneration due to the intrinsic environmental inhibition. Although stem cell therapy seems to be a promising approach towards nerve repair, it is essential to use the distinct three-dimensional architecture of a cell scaffold with proper biomolecule embedding in order to ensure that the local environment can be controlled well enough for growth and survival. Many approaches have been developed for the fabrication of 3D scaffolds, and more recently, fiber-based scaffolds produced via the electrospinning have been garnering increasing interest, as it offers the opportunity for control over fiber composition, as well as fiber mesh porosity using a relatively simple experimental setup. All these attributes make electrospun fibers a new class of promising scaffolds for neural tissue engineering. Therefore, the purpose of this doctoral study is to investigate the use of the novel material PGD and its derivative PGDF for obtaining fiber scaffolds using the electrospinning. The performance of these scaffolds, combined with neural lineage cells derived from ESCs, was evaluated by the dissolvability test, Raman spectroscopy, cell viability assay, real time PCR, Immunocytochemistry, extracellular electrophysiology, etc. The newly designed collector makes it possible to easily obtain fibers with adequate length and integrity. The utilization of a solvent like ethanol and water for electrospinning of fibrous scaffolds provides a potentially less toxic and more biocompatible fabrication method. Cell viability testing demonstrated that the addition of gelatin leads to significant improvement of cell proliferation on the scaffolds. Both real time PCR and Immunocytochemistry analysis indicated that motor neuron differentiation was achieved through the high motor neuron gene expression using the metabolites approach. The addition of Fumaric acid into fiber scaffolds further promoted the differentiation. Based on the results, this newly fabricated electrospun fiber scaffold, combined with neural lineage cells, provides a potential alternate strategy for nerve injury repair.^

Assessment of the Occurrence and Potential Risks of Antibiotics and their Metabolites in South Florida Waters Using Liquid Chromatography Tandem Mass Spectrometry

An automated on-line SPE-LC-MS/MS method was developed for the quantitation of multiple classes of antibiotics in environmental waters. High sensitivity in the low ng/L range was accomplished by using large volume injections with 10-mL of sample. Positive confirmation of analytes was achieved using two selected reaction monitoring (SRM) transitions per antibiotic and quantitation was performed using an internal standard approach. Samples were extracted using online solid phase extraction, then using column switching technique; extracted samples were immediately passed through liquid chromatography and analyzed by tandem mass spectrometry. The total run time per each sample was 20 min. The statistically calculated method detection limits for various environmental samples were between 1.2 and 63 ng/L. Furthermore, the method was validated in terms of precision, accuracy and linearity. The developed analytical methodology was used to measure the occurrence of antibiotics in reclaimed waters (n=56), surface waters (n=53), ground waters (n=8) and drinking waters (n=54) collected from different parts of South Florida. In reclaimed waters, the most frequently detected antibiotics were nalidixic acid, erythromycin, clarithromycin, azithromycin trimethoprim, sulfamethoxazole and ofloxacin (19.3-604.9 ng/L). Detection of antibiotics in reclaimed waters indicates that they can’t be completely removed by conventional wastewater treatment process. Furthermore, the average mass loads of antibiotics released into the local environment through reclaimed water were estimated as 0.248 Kg/day. Among the surface waters samples, Miami River (reaching up to 580 ng/L) and Black Creek canal (up to 124 ng/L) showed highest concentrations of antibiotics. No traces of antibiotics were found in ground waters. On the other hand, erythromycin (monitored as anhydro erythromycin) was detected in 82% of the drinking water samples (n.d-66 ng/L). The developed approach is suitable for both research and monitoring applications. Major metabolites of antibiotics in reclaimed wates were identified and quantified using high resolution benchtop Q-Exactive orbitrap mass spectrometer. A phase I metabolite of erythromycin was tentatively identified in full scan based on accurate mass measurement. Using extracted ion chromatogram (XIC), high resolution data-dependent MS/MS spectra and metabolic profiling software the metabolite was identified as desmethyl anhydro erythromycin with molecular formula C36H63NO12 and m/z 702.4423. The molar concentration of the metabolite to erythromycin was in the order of 13 %. To my knowledge, this is the first known report on this metabolite in reclaimed water. Another compound acetyl-sulfamethoxazole, a phase II metabolite of sulfamethoxazole was also identified in reclaimed water and mole fraction of the metabolite represent 36 %, of the cumulative sulfamethoxazole concentration. The results were illustrating the importance to include metabolites also in the routine analysis to obtain a mass balance for better understanding of the occurrence, fate and distribution of antibiotics in the environment. Finally, all the antibiotics detected in reclaimed and surface waters were investigated to assess the potential risk to the aquatic organisms. The surface water antibiotic concentrations that represented the real time exposure conditions revealed that the macrolide antibiotics, erythromycin, clarithromycin and tylosin along with quinolone antibiotic, ciprofloxacin were suspected to induce high toxicity to aquatic biota. Preliminary results showing that, among the antibiotic groups tested, macrolides posed the highest ecological threat, and therefore, they may need to be further evaluated with, long-term exposure studies considering bioaccumulation factors and more number of species selected. Overall, the occurrence of antibiotics in aquatic environment is posing an ecological health concern.

A longitudinal study of Project Trio: A dropout prevention program in a large urban environment

The subject of dropout prevention/reduction is deservedly receiving attention as a problem that, if not resolved, could threaten our national future.^ This study investigates a small segment of the overall dropout problem, which has apparently unique features of program design and population selection. The evidence presented here should add to the knowledge bank of this complicated problem.^ Project Trio was one of a number of dropout prevention programs and activities which were conducted in Dade County school years 1984-85 and 1985-86, and it is here investigated longitudinally through the end of the 1987-88 school year. It involved 17 junior and senior high schools, and 27 programs, 10 the first year and 17 the second, with over 1,000 total students, who had been selected by the schools from a list of the "at risk" students provided by the district, and were divided approximately evenly into the classical research design of an experimental group and the control group, which following standard procedure was to take the regular school curriculum. No school had more than 25 students in either group.^ Each school modified the basic design of the project to accommodate the individual school characteristics and the perceived needs of their students; however all schools projects were to include some form of academic enhancement, counseling and career awareness study.^ The conclusion of this study was that the control group had a significantly lower dropout rate than the experimental group. Though impossible to make a certain determination of the reasons for this unexpected result, it appears from evidence presented that one cause may have been inadequate administration at the local level.^ This study was also a longitudinal investigation of the "at risk" population as a whole for the three and four year period, to determine if academic factors were present in records may be used to identify dropout proneness.^ A significant correlation was found between dropping out and various measures including scores on the Quality of School Life Instrument, attendance, grade point averages, mathematics grades, and overage in grade, important identifiers in selection for dropout prevention programs. ^

Development of an odor management plan for industrial stationary sources in an urban environment: A case study for Broward County, Florida

Objectionable odors remain at the top of air pollution complaints in urban areas such as Broward County that is subject to increasing residential and industrial developments. The odor complaints in Broward County escalated by 150 percent for the 2001 to 2004 period although the population increased by only 6 percent. It is estimated that in 2010 the population will increase to 2.5 million. Relying solely on enforcing the local odor ordinance is evidently not sufficient to manage the escalating odor complaint trends. An alternate approach similar to odor management plans (OMPs) that are successful in managing major malodor sources such as animal farms is required. ^ This study aims to develop and determine the feasibility of implementing a comprehensive odor management plan (COMP) for the entire Broward County. Unlike existing OMPs for single sources where the receptors (i.e. the complainants) are located beyond the boundary of the source, the COMP addresses a complex model of multiple sources and receptors coexisting within the boundary of the entire county. Each receptor is potentially subjected to malodor emissions from multiple sources within the county. Also, the quantity and quality of the source/receptor variables are continuously changing. ^ The results of this study show that it is feasible to develop a COMP that adopts a systematic procedure to: (1) Generate maps of existing odor complaint areas and malodor sources, (2) Identify potential odor sources (target sources) responsible for existing odor complaints, (3) Identify possible odor control strategies for target sources, (4) Determine the criteria for implementing odor control strategies, (5) Develop an odor complaint response protocol, and (6) Conduct odor impact analyses for new sources to prevent future odor related issues. Geographic Information System (GIS) is used to identify existing complaint areas. A COMP software that incorporates existing United States Environmental Protection Agency (EPA) air dispersion software is developed to determine the target sources, predict the likelihood of new complaints, and conduct odor impact analysis. The odor response protocol requires pre-planning field investigations and conducting surveys to optimize the local agency available resources while protecting the citizen's welfare, as required by the Clean Air Act. ^

Attitudes towards the marine environment and implications for marine resource management in Seward, Alaska

This research, conducted in 2006-2008, examines the ways in which various groups involved with the marine resources of Seward, Alaska construct attitudes towards the environment. Participant observation and semi-structured interviews are used to assess how commercial halibut fishers, tour boat operators, local residents and government officials understand the marine environment based on their previous experiences. This study also explores how ideologies relate to the current practices of each group. Two theories orient the analyses: The first, cultural modeling provided a theoretical and methodological framework for pursuing a more comprehensive analysis of resource management. The second, Theory of Reasoned Action (Ajzen and Fishbein 1980), guided the analysis of the ways in which each participant’s ideology towards the marine environment relates to their practice. Aside from contributing to a better understanding of a coastal community’s ideologies and practices, this dissertation sought to better understand the role of ecological ideologies and behaviors in fisheries management. The research illustrates certain domains where ideologies and practices concerning Pacific halibut and the marine environment differ among commercial fishers, government, and management officials, tour boat operators and residents of Seward, AK. These differences offer insights into how future collaborative efforts between government officials, managers and local marine resource users might better incorporate local ideology into management, and provide ecological information to local marine resource users in culturally appropriate ways.

Historical analysis of two contemporary issues at the International Community School of Abidjan, Cote d'Ivoire: Local -hired and overseas -hired teacher relationships. Community perceptions of the U.S. Embassy at ICSA

This study was a qualitative investigation to ascertain and describe two of the current issues at the International Community School of Abidjan, examine their historical bases, and analyze their impact on the school environment.^ Two issues emerged during the inquiry phase of this study: (1) the relationship between local-hired and overseas-hired teachers in light of the January 1994 devaluation which polarized the staff by negating a four-year salary scale that established equity, (2) the school community's wide variance in the perceived power that the U.S. Embassy has on school operations based on its role as ICSA's founding sponsor.^ A multiple studies approach was used in gathering data. An extensive examination of the school's archives was used to reconstruct an historical overview of ICSA. An initial questionnaire was distributed to teachers and administrators at an educational conference to determine the scope of the 1994 devaluation of the West and Central African CFA and its impact on school personnel in West African American-sponsored overseas schools (ASOS). Personal interviews were conducted with the school staff, administration, school board members, and relevant historical participants to determine the principal issues at ICSA at that time. The researcher, an overseas-hired teacher, also used participant observations to collect data. Findings based on these sources were used to analyze the two issues from an historical perspective and to form conclusions.^ Findings in this study pertaining to the events induced by the French and African governments' decision to implement a currency devaluation in January 1994 were presented in ex post-facto chronological narrative form to describe the events which transpired, describe the perception of school personnel involved in these events, examine the final resolution and interpret these events within a historical framework for analysis.^ The topic of the U.S. Embassy and its role at ICSA emerged inductively from open-ended personal interviews conducted over the course of a year. Contradictory perspectives were examined and researched for accuracy and cause. The results of this inquiry presented the U.S. Embassy role at ICSA from a two-sided perspective, examined the historical role of the Embassy, and presented means by which the role and responsibility of the U.S. Embassy could best be communicated to the school community.^ The final chapter provides specific actions for mediation of problems stemming from these issues, implications for administrators and teachers currently involved in overseas schools or considering the possibility, and suggestions for future inquiries.^ Examination of a two-tier salary scale for local-hired and overseas-hired teachers generated the following recommendations: movement towards a single salary scale when feasible, clearly stated personnel policies and full disclosure of benefits, a uniform certification standard, professional development programs and awareness of the impact of this issue on staff morale.^ Divergent perceptions and attitudes toward the role of the U.S. Embassy produced these recommendations: a view towards limiting the number of Americans on ASOS school boards, open school board meetings, selection of Embassy Administrative Officers who can educate school communities on the exact role of the Embassy, educating parents through the outreach activities that communicate American educational philosophy and involve all segments of the international community, and a firm effort on the part of the ASOS to establish the school's autonomy from special interests. ^

Cultural perceptions of space and place in the South Florida *environment

Public opinion polls in the United States reveal that a great majority of Americans are aware and show concern about ecological issues and the need to preserve natural areas. In South Florida, natural resources have been subjected to enormous strain as the pressure to accommodate a growing population has led to rapid development. Suburbs have been built on areas that were once natural wetlands and farmlands, and the impact today shows a landscape where natural places have all but disappeared. This dissertation examines the intersection between the perceptions that individuals living in the South Florida region have with respect to the natural environment and local ecological problems with where their relationship to nature takes place. ^ The research is based upon both quantitative and qualitative data. The principal methodology used in this research is the ethnographic method, which employed the data gathering techniques of in-depth interviewing and participant observation. The objective of the qualitative portion of the study was to determine how people perceive and relate to their immediate environment. The quantitative portion of the study employed telephone survey data from the FIU/Florida Poll 2000. Data collected through this survey provided the basis to statistically test responses to the research questions. ^ The findings show that people in South Florida have a general idea of the relationship between the human population and the environment but very little knowledge of how they individually affect each other. The experience of private places and public spaces in everyday life permits people to compartmentalize cultural values and understandings of the natural world in separate cognitive schemas. The appreciation of the natural world has almost no connection to their personal sense of obligation to preserve the environment. That obligation is only felt in their home space even though the South Florida environment overall struggles desperately with water shortages, land encroachment, and a rapidly expanding human population whose activities continuously aggravate an already delicate natural balance. ^

The Influence of Servicescape and Local Food Attributes on Pleasure and Revisit Intention in an Upscale-Casual Dining Restaurant

The current study looks at the relationship between servicescape, emotional product involvement, perceived quality of local foods, the positive emotion of pleasure, and revisit intention in an upscale buffet style restaurant on a university campus in the Southeastern U.S. Test results show positive relationships between all of the constructs in the proposed conceptual model. The study also gives practitioners and academics insights into practices that can help to market the use of local foods through the restaurant environment in order to engage emotionally involved customers. This marketing can illicit pleasurable feelings and increase perceived product quality of local foods with the purpose of getting customers to revisit the restaurant. Suggestions for further research on the subject are proposed.

Attitudes towards the Marine Environment and Implications for Marine Resource Management in Seward, Alaska

This research, conducted in 2006-2008, examines the ways in which various groups involved with the marine resources of Seward, Alaska construct attitudes towards the environment. Participant observation and semi-structured interviews are used to assess how commercial halibut fishers, tour boat operators, local residents and government officials understand the marine environment based on their previous experiences. This study also explores how ideologies relate to the current practices of each group. Two theories orient the analyses: The first, cultural modeling provided a theoretical and methodological framework for pursuing a more comprehensive analysis of resource management. The second, Theory of Reasoned Action (Ajzen and Fishbein 1980), guided the analysis of the ways in which each participant’s ideology towards the marine environment relates to their practice. Aside from contributing to a better understanding of a coastal community’s ideologies and practices, this dissertation sought to better understand the role of ecological ideologies and behaviors in fisheries management. The research illustrates certain domains where ideologies and practices concerning Pacific halibut and the marine environment differ among commercial fishers, government, and management officials, tour boat operators and residents of Seward, AK. These differences offer insights into how future collaborative efforts between government officials, managers and local marine resource users might better incorporate local ideology into management, and provide ecological information to local marine resource users in culturally appropriate ways.

Analysis of Land Use Change as a Method of Predicting Water Demands in an Urbanizing Environment: Redland, Miami-Dade County, Florida

The fluctuation in water demand in the Redland community of Miami-Dade County was examined using land use data from 2001 and 2011 and water estimation techniques provided by local and state agencies. The data was converted to 30 m mosaicked raster grids that indicated land use change, and associated water demand measured in gallons per day per acre. The results indicate that, first, despite an increase in population, water demand decreased overall in Redland from 2001 to 2011. Second, conversion of agricultural lands to residential lands actually caused a decrease in water demand in most cases while acquisition of farmland by public agencies also caused a sharp decline. Third, conversion of row crops and groves to nurseries was substantial and resulted in a significant increase in water demand in all such areas converted. Finally, estimating water demand based on land use, rather than population, is a more accurate approach.