Preimplantation genetic diagnosis: Analysis of gene expression, nuclear DNA and cytoplasmic DNA

Preimplantation genetic diagnosis (PGD) following in vitro fertilization (IVF) offers couples at risk for transmitting genetic disorders the opportunity to identify affected embryos prior to replacement. In particular, embryo gender determination permits screening for X-linked diseases of unknown etiology. Analysis of embryos can be performed by polymerase chain reaction (PCR) amplification of material obtained by micromanipulation. This approach provides an alternative to the termination of an established pregnancy following chorionic villi sampling or amniocentesis. ^ Lately, the focus of preimplantation diagnosis and intervention has been shifting toward an attempt to correct cytoplasmic deficiencies. Accordingly, it is the aim of this investigation to develop methods to permit the examination of single cells or components thereof for clinical evaluation. In an attempt to lay the groundwork for precise therapeutic intervention for age related aneuploidy, transcripts encoding proteins believed to be involved in the proper segregation of chromosomes during human oocyte maturation were examined and quantified. Following fluorescent rapid cycle RT-PCR analysis it was determined that the concentration of cell cycle checkpoint gene transcripts decreases significantly as maternal age increases. Given the well established link between increasing maternal age and the incidence of aneuploidy, these results suggest that the degradation of these messages in aging oocytes may be involved with inappropriate chromosome separation during meiosis. ^ In order to investigate the cause of embryonic rescue observed following clinical cytoplasmic transfer procedures and with the objective of developing a diagnostic tool, mtDNA concentrations in polar bodies and subcellular components were evaluated. First, the typical concentration of mtDNA in human and mouse oocytes was determined by fluorescent rapid cycle PCR. Some disparity was noted between the copy numbers of individual cytoplasmic samples which may limit the use of the current methodology for the clinical assessment of the corresponding oocyte. ^

Dean of the school board: An oral history of the distinguished career of G. Holmes Braddock

Effective school board leadership is often an ephemeral ideal in today's highly politicized public education arena. However, effective leadership is necessary in order to ensure a fair and equitable education for all students. This dissertation described and explained one school board member's perspective of his career as a lens from which to view and assess public educational policy making in Miami-Dade County. ^ Now retired after thirty-eight years of service, G. Holmes Braddock is the longest serving, contemporary, urban school board member in the country. Spanning nearly four decades, his perspective provides a comprehensive view of urban education both locally and nationally. The significance of his longevity and the impact of his leadership on educational policy-making was the focus of indepth interviews with Mr. Braddock and other key educational “influentials.” From this transcript data, recurring themes were revealed and categorized. Five elements of his perspective, i.e., teacher professionalization; desegregation; athletics; bilingual education; and his comprehensive leadership role, were identified and analyzed, as were five variables of his perspective, i.e., fairness; integrity; honesty; courage; and the situational context. Other secondary source material, such as excerpts from newspaper articles, school board minutes, and items from Mr. Braddock's own personal effects further augmented and triangulated the data. ^ Given that the purpose of this study was to describe and explain Mr. Braddock's perspective of his school board career, the findings can be understood from two different viewpoints. The elements of Mr. Braddock's perspective describe or characterize his career and represent the significant policy issues in which he demonstrated exceptional vision and leadership. However, taken alone, these elements cannot fully explain his distinguished career. Rather, an analysis of the variables of Mr. Braddock's perspective provides an explanation for the effectiveness of his leadership role. Personality traits such as fairness, integrity, honesty and courage and the impact of the situational context were factors that strongly influenced Mr. Braddock's decision-making. Thus, Mr. Braddock's school board career can be holistically understood as the intersection of person, place and time with significant public education policy issues. ^ The results of this study provide a unique and historical perspective of school board politics in Miami-Dade County. From Mr. Braddock's perspective, we are able to view one individual's leadership role over time and its impact on local public education policy. ^

Improved sampling and detection of ignitable liquid residues and explosives by mass spectrometry and ion mobility spectrometry

Fire debris evidence is submitted to crime laboratories to determine if an ignitable liquid (IL) accelerant was used to commit arson. An ignitable liquid residue (ILR) may be difficult to analyze due to interferences, complex matrices, degradation, and low concentrations of analytes. Debris from an explosion and pre-detonated explosive compounds are not trivial to detect and identify due to sampling difficulties, complex matrices, and extremely low amounts (nanogram) of material present. The focus of this research is improving the sampling and detection of ILR and explosives through enhanced sensitivity, selectivity, and field portable instrumentation. Solid Phase MicroExtraction (SPME) enhanced the extraction of ILR by two orders of magnitude over conventional activated charcoal strip (ACS) extraction. Gas chromatography tandem mass spectrometry (GC/MS/MS) improved sensitivity of ILR by one order of magnitude and explosives by two orders of magnitude compared to gas chromatography mass spectrometry (GC/MS). Improvements in sensitivity were attributed to enhanced selectivity. An interface joining SPME to ion mobility spectrometry (IMS) has been constructed and evaluated to improve field detection of hidden explosives. The SPME-IMS interface improved the detection of volatile and semi-volatile explosive compounds and successfully adapted the IMS from a particle sampler into a vapor sampler. ^

The effects of a General Instructional Strategies course on the utilization of effective instructional strategies by teachers

The focus of this study was to explain the extent to which theoretically effective teaching strategies taught in a course on generic instructional strategies are being implemented by teachers in their actual teaching practice. ^ A multivariate causal-comparative (ex-post-facto) design was used to answer the research question. A teacher observation protocol, the General Instructional Strategies Analysis (GISA) was constructed and used to assess the utilization of instructional strategies in the classroom. The data of this study also included open-ended field notes taken during observations. ^ Multivariate Analyses of Variance (MANOVA) was used to compare the teaching strategies (set, effective explanation, hands-on activity, cooperative learning activity, higher order questioning, closure) of the group who had taken a general instructional strategies course (N=36) and the group who had not (N=36). Results showed a statistically significant difference between the two groups. The group who had taken the course implemented these strategies more effectively in almost all categories of effective teaching. Follow-up univariate tests of the dependent variables showed significant differences between the two groups in five of the six areas (hands-on activity being an exception). A second MANOVA compared the two groups on the effective use of attending behaviors (teacher movement/eye contact/body language/physical space, brief verbal acknowledgements/voice inflection/modulation/pitch, use of visuals, prompting/probing, praise/feedback/rewards, wait-time I and II). Results also showed a multivariate difference between the two groups. Follow-up univariate tests on the related dependent variables showed that five of the six were significantly different between the two groups. The group who had taken the course implemented the strategies more effectively. An analysis of the field notes provided further evidence regarding the pervasiveness of these differences between the teaching practices of the two groups. ^ It was concluded that taking a course in general instructional strategies increases the utilization of effective strategies in the classroom by teachers. ^

Evaluation of contact and non-contact trapping efficiencies of human scent chemical profiles and their stabilities under different environmental conditions

Establishing an association between the scent a perpetrator left at a crime scene to the odor of the suspect of that crime is the basis for the use of human scent identification evidence in a court of law. Law enforcement agencies gather evidence through the collection of scent from the objects that a perpetrator may have handled during the execution of the criminal act. The collected scent evidence is consequently presented to the canines for identification line-up procedures with the apprehended suspects. Presently, canine scent identification is admitted as expert witness testimony, however, the accurate behavior of the dogs and the scent collection methods used are often challenged by the court system. The primary focus of this research project entailed an evaluation of contact and non-contact scent collection techniques with an emphasis on the optimization of collection materials of different fiber chemistries to evaluate the chemical odor profiles obtained using varying environment conditions to provide a better scientific understanding of human scent as a discriminative tool in the identification of suspects. The collection of hand odor from female and male subjects through both contact and non-contact sampling approaches yielded new insights into the types of VOCs collected when different materials are utilized, which had never been instrumentally performed. Furthermore, the collected scent mass was shown to be obtained in the highest amounts for both gender hand odor samples on cotton sorbent materials. Compared to non-contact sampling, the contact sampling methods yielded a higher number of volatiles, an enhancement of up to 3 times, as well as a higher scent mass than non-contact methods by more than an order of magnitude. The evaluation of the STU-100 as a non-contact methodology highlighted strong instrumental drawbacks that need to be targeted for enhanced scientific validation of current field practices. These results demonstrated that an individual's human scent components vary considerably depending on the method used to collect scent from the same body region. This study demonstrated the importance of collection medium selection as well as the collection method employed in providing a reproducible human scent sample that can be used to differentiate individuals.

Deprofessionalization of curriculum decision -making

The purpose of this study was to analyze the evolution of Florida state level policy efforts and to assess the responding educational policy development and implementation at the local school district level. The focus of this study was the secondary language arts curriculum in Miami-Dade County Public Schools. ^ Data was collected using document analysis as a source of meaning making out of the language sets proffered by agencies at each level. A matrix was created based on Klein's levels of curriculum decision-making and Functional Process Theory categories of policy formation. The matrix allowed the researcher to code and classify specific information in terms accountability/high-stakes testing; authority; outside influences; and operational/structural organization. ^ Federal policy documents provided a background and impetus for much of what originated at the State level. The State then produced policy directives which were accepted by the District and specific policy directives and guidelines for practice. No evidence was found indicating the involvement of any other agencies in the development, transmission or implementation of the State level initiated policies. ^ After analyzing the evolutionary process, it became clear that state policy directives were never challenged or discussed. Rather, they were accepted as standards to be met and as such, school districts complied. Policy implementation is shown to be a top-down phenomenon. No evidence was found indicating a dialogue between state and local systems, rather the state, as the source of authority, issued specifically worded policy directives and the district complied. Finally, this study recognizes that outside influences play an important role in shaping the education reform policy in the state of Florida. The federal government, through NCLB and other initiatives created a climate which led almost naturally to the creation of the Florida A+ Plan. Similarly, the concern of the business community, always interested in the production of competent workers, continued to support efforts at raising the minimum skill level of Florida high school graduates. ^ Suggestions are made for future research including the examination of local school sites in order to assess the overall nature of the school experience rather than rely upon performance indicators mandated by state policy. ^

Predicting the in-term persistence of community college English-as-a-second-language students

The English-as-a-second-language (ESL) community college student population has increased notably in the past decade, but a decreasing number of these students are completing courses, programs, or degrees (Erisman & Looney, 2008). These students come to college with unique background experiences, and once in college, deal with challenging linguistic, academic, and social integration issues. Though they are not linguistically homogenous, and they do not have a common purpose, ESL students share the common goal of attending community college to learn to speak English (Szelényi & Chang, 2002). Course completion is a primary measure of progress toward that goal, and is therefore an issue of concern for both ESL students and community colleges, which continue to be the access point for language-minority students progressing into higher education (Laden, 2004).^ The purpose of this study was to investigate the factors that predict in-term persistence of community college ESL students. A mixed methods research design consisting of two phases was utilized, and participants in this study were ESL students enrolled in a large community college in south Florida. Phase 1 students completed the Community College ESL Student Questionnaire (CCSEQ), which collected demographic data and data on entry characteristics, academic integration, and social integration. Discriminant and descriptive analyses were used to report the data collected in Phase I. Phase 2 students were a matching cohort of completing and non-completing students who participated in semi-structured interviews at the end of the term. Data collected in the interviews were analyzed thematically, using a constant comparative method as described by Glaser and Strauss (1967).^ Students’ self reported demographic data, background characteristics, goal commitment, and integration factors on the CCSEQ showed no significance between the students who completed the term and the students who did not complete the term. However, several differentiating themes emerged from the interview data, which indicated differences in goal commitment and integration between the two groups. The focus of non-completers on getting good grades rather than completing the course, and the commitment of completers to the goal of finishing the class in order to go forward, both raise questions for future research studies.^

The Caribbean Court of Justice: An investigation of the impact of elites on its creation and present structure as it relates to original jurisdiction

The primary focus of this dissertation is to determine the degree to which political, economic, and socio-cultural elites in Jamaica and Trinidad & Tobago influenced the development of the Caribbean Court of Justice's (CCJ) original jurisdiction. As members of the Caribbean Community (CARICOM), both states replaced their protectionist model with open regionalism at the end of the 1980s. Open regionalism was adopted to make CARICOM member states internationally competitive. Open regionalism was also expected to create a stable regional trade environment. To ensure a stable economic environment, a regional court with original jurisdiction was proposed. A six member Preparatory Committee on the Caribbean Court of Justice (PREPCOM), on which Jamaica and Trinidad & Tobago sat, was formed to draft the Agreement Establishing the Caribbean Court of Justice that would govern how the Court would interpret the Revised Treaty of Chaguaramas (RTC) and enforce judgments. ^ Through the use of qualitative research methods, namely elite interviews, document data, and text analysis, and a focus on three levels of analysis, that is, the international, regional, and domestic, three major conclusions are drawn. First, changes in the international economic environment caused Jamaica and Trinidad & Tobago to support the establishment of a regional court. Second, Jamaica had far greater influence on the final structure of the CCJ than Trinidad & Tobago. Third, it was found that in both states the political elite had the greatest influence on the development and structure of the CCJ. The economic elite followed by the socio-cultural elite were found to have a lesser impact. These findings are significant because they account for the impact of elites and elite behavior on institutions in a much-neglected category of states: the developing world.^

Boundary : an exploration of architecture's power to mark a boundary

Cities are no longer recognizing their boundaries beyond their legal or political implications. This thesis explored the architecture of a boundary in the design of a structure that visually and symbolically marked the current Miami-Dade County development limit along Krome Avenue. This limit ultimately separates the city from the endangered Everglades ecosystem. Through the examination of ancient boundaries such as Medieval and Renaissance Military fortifications, as well as contemporary interventions such as Steven Holl's "Edge of the City Projects", an architectural vocabulary was developed considering the Double Wall, Bastion, Rampart, and Gate principles. However, rather than considering these principles to defend the city from outside forces, the focus of this exploration is their inversion in order to contain the city by strengthening its periphery visually and symbolically. From this exploration, it was concluded that a successful boundary or limit should be visible and significant. Furthermore, it should serve as an informative and reflective landmark.

Deprofessionalization of curriculum decision-making

The purpose of this study was to analyze the evolution of Florida state level policy efforts and to assess the responding educational policy development and implementation at the local school district level. The focus of this study was the secondary language arts curriculum in Miami-Dade County Public Schools. Data was collected using document analysis as a source of meaning making out of the language sets proffered by agencies at each level. A matrix was created based on Klein's levels of curriculum decision-making and Functional Process Theory categories of policy formation. The matrix allowed the researcher to code and classify specific information in terms accountability/high-stakes testing; authority; outside influences; and operational/structural organization. Federal policy documents provided a background and impetus for much of what originated at the State level. The State then produced policy directives which were accepted by the District and specific policy directives and guidelines for practice. No evidence was found indicating the involvement of any other agencies in the development, transmission or implementation of the State level initiated policies. After analyzing the evolutionary process, it became clear that state policy directives were never challenged or discussed. Rather, they were accepted as standards to be met and as such, school districts complied. Policy implementation is shown to be a top-down phenomenon. No evidence was found indicating a dialogue between state and local systems, rather the state, as the source of authority, issued specifically worded policy directives and the district complied. Finally, this study recognizes that outside influences play an important role in shaping the education reform policy in the state of Florida. The federal government, through NCLB and other initiatives created a climate which led almost naturally to the creation of the Florida A+ Plan. Similarly, the concern of the business community, always interested in the production of competent workers, continued to support efforts at raising the minimum skill level of Florida high school graduates. Suggestions are made for future research including the examination of local school sites in order to assess the overall nature of the school experience rather than rely upon performance indicators mandated by state policy.

The international legal thought of Carl Schmitt : towards a critique of the contemporary international order

The aim of the thesis is to develop a critique of current liberal conceptualizations of international order. In order to conduct this critique, this thesis revisits the arguments first put forth by the German legal and political theorist Carl Schmitt. Schmitt conceptualizes a tripartite unity between law, order, and place. This unity, established at the constituent moment of land-appropriation, forms a concrete nomos, which subsequently creates the contours of the legal and political order. The establishment of the concrete order is necessarily the construction of a territorial boundary that designates an inside and an outside of the polity. By speaking of a nomos of the earth, Schmitt globalized this understanding of concrete order by looking at the various historical developments that created a "line" between the concrete applicability of interstate norms and a region where the exceptional situation prevails. The critique presented in this thesis is concerned with the lack of concrete boundary conditions within the current international legal order. It is argued that this lack of a well-defined boundary condition is what results in extreme forms of violence that were traditionally bracketed.

Design and implementation of a relative localization system for ground and aerial robotic teams

The main focus of this thesis is to address the relative localization problem of a heterogenous team which comprises of both ground and micro aerial vehicle robots. This team configuration allows to combine the advantages of increased accessibility and better perspective provided by aerial robots with the higher computational and sensory resources provided by the ground agents, to realize a cooperative multi robotic system suitable for hostile autonomous missions. However, in such a scenario, the strict constraints in flight time, sensor pay load, and computational capability of micro aerial vehicles limits the practical applicability of popular map-based localization schemes for GPS denied navigation. Therefore, the resource limited aerial platforms of this team demand simpler localization means for autonomous navigation. Relative localization is the process of estimating the formation of a robot team using the acquired inter-robot relative measurements. This allows the team members to know their relative formation even without a global localization reference, such as GPS or a map. Thus a typical robot team would benefit from a relative localization service since it would allow the team to implement formation control, collision avoidance, and supervisory control tasks, independent of a global localization service. More importantly, a heterogenous team such as ground robots and computationally constrained aerial vehicles would benefit from a relative localization service since it provides the crucial localization information required for autonomous operation of the weaker agents. This enables less capable robots to assume supportive roles and contribute to the more powerful robots executing the mission. Hence this study proposes a relative localization-based approach for ground and micro aerial vehicle cooperation, and develops inter-robot measurement, filtering, and distributed computing modules, necessary to realize the system. The research study results in three significant contributions. First, the work designs and validates a novel inter-robot relative measurement hardware solution which has accuracy, range, and scalability characteristics, necessary for relative localization. Second, the research work performs an analysis and design of a novel nonlinear filtering method, which allows the implementation of relative localization modules and attitude reference filters on low cost devices with optimal tuning parameters. Third, this work designs and validates a novel distributed relative localization approach, which harnesses the distributed computing capability of the team to minimize communication requirements, achieve consistent estimation, and enable efficient data correspondence within the network. The work validates the complete relative localization-based system through multiple indoor experiments and numerical simulations. The relative localization based navigation concept with its sensing, filtering, and distributed computing methods introduced in this thesis complements system limitations of a ground and micro aerial vehicle team, and also targets hostile environmental conditions. Thus the work constitutes an essential step towards realizing autonomous navigation of heterogenous teams in real world applications.

E-service quality:development of a hierarchical model

Using survey data from 358 online customers, the study finds that the e-service quality construct conforms to the structure of a third-order factor model that links online service quality perceptions to distinct and actionable dimensions, including (1) website design, (2) fulfilment, (3) customer service, and (4) security/privacy. Each dimension is found to consist of several attributes that define the basis of e-service quality perceptions. A comprehensive specification of the construct, which includes attributes not covered in existing scales, is developed. The study contrasts a formative model consisting of 4 dimensions and 16 attributes against a reflective conceptualization. The results of this comparison indicate that studies using an incorrectly specified model overestimate the importance of certain e-service quality attributes. Global fit criteria are also found to support the detection of measurement misspecification. Meta-analytic data from 31,264 online customers are used to show that the developed measurement predicts customer behavior better than widely used scales, such as WebQual and E-S-Qual. The results show that the new measurement enables managers to assess e-service quality more accurately and predict customer behavior more reliably.

From ‘poor parenting’ to micro-management:coalition governance and the sponsorship of arm’s-length bodies in the United Kingdom, 2010–13

The delegation of public tasks to arm’s-length bodies remains a central feature of contemporary reform agendas within both developed and developing countries. The role and capacity of political and administrative principals (i.e. ministers and departments of state) to control the vast network of arm’s-length bodies for which they are formally responsible is therefore a critical issue within and beyond academe. In the run-up to the 2010 General Election in the United Kingdom, the ‘quango conundrum’ emerged as an important theme and all three major parties committed themselves to shift the balance of power back towards ministers and sponsor departments. This article presents the results of the first major research project to track and examine the subsequent reform process. It reveals a stark shift in internal control relationships from the pre-election ‘poor parenting’ model to a far tighter internal situation that is now the focus of complaints by arm’s-length bodies of micro-management. This shift in the balance of power and how it was achieved offers new insights into the interplay between different forms of governance and has significant theoretical and comparative relevance. Points for practitioners: For professionals working in the field of arm’s-length governance, the article offers three key insights. First, that a well-resourced core executive is critical to directing reform given the challenges of implementing reform in a context of austerity. Second, that those implementing reform will also need to take into account the diverse consequences of centrally imposed reform likely to result in different departments with different approaches to arm’s-length governance. Third, that reforming arm’s-length governance can affect the quality of relationships, and those working in the field will need to mitigate these less tangible challenges to ensure success.

Soundscape Ecology of Hawaiian Spinner Dolphin Resting Bays

Sound is a key sensory modality for Hawaiian spinner dolphins. Like many other marine animals, these dolphins rely on sound and their acoustic environment for many aspects of their daily lives, making it is essential to understand soundscape in areas that are critical to their survival. Hawaiian spinner dolphins rest during the day in shallow coastal areas and forage offshore at night. In my dissertation I focus on the soundscape of the bays where Hawaiian spinner dolphins rest taking a soundscape ecology approach. I primarily relied on passive acoustic monitoring using four DSG-Ocean acoustic loggers in four Hawaiian spinner dolphin resting bays on the Kona Coast of Hawai‛i Island. 30-second recordings were made every four minutes in each of the bays for 20 to 27 months between January 8, 2011 and March 30, 2013. I also utilized concomitant vessel-based visual surveys in the four bays to provide context for these recordings. In my first chapter I used the contributions of the dolphins to the soundscape to monitor presence in the bays and found the degree of presence varied greatly from less than 40% to nearly 90% of days monitored with dolphins present. Having established these bays as important to the animals, in my second chapter I explored the many components of their resting bay soundscape and evaluated the influence of natural and human events on the soundscape. I characterized the overall soundscape in each of the four bays, used the tsunami event of March 2011 to approximate a natural soundscape and identified all loud daytime outliers. Overall, sound levels were consistently louder at night and quieter during the daytime due to the sounds from snapping shrimp. In fact, peak Hawaiian spinner dolphin resting time co-occurs with the quietest part of the day. However, I also found that humans drastically alter this daytime soundscape with sound from offshore aquaculture, vessel sound and military mid-frequency active sonar. During one recorded mid-frequency active sonar event in August 2011, sound pressure levels in the 3.15 kHz 1/3rd-octave band were as high as 45.8 dB above median ambient noise levels. Human activity both inside (vessels) and outside (sonar and aquaculture) the bays significantly altered the resting bay soundscape. Inside the bays there are high levels of human activity including vessel-based tourism directly targeting the dolphins. The interactions between humans and dolphins in their resting bays are of concern; therefore, my third chapter aimed to assess the acoustic response of the dolphins to human activity. Using days where acoustic recordings overlapped with visual surveys I found the greatest response in a bay with dolphin-centric activities, not in the bay with the most vessel activity, indicating that it is not the magnitude that elicits a response but the focus of the activity. In my fourth chapter I summarize the key results from my first three chapters to illustrate the power of multiple site design to prioritize action to protect Hawaiian spinner dolphins in their resting bays, a chapter I hope will be useful for managers should they take further action to protect the dolphins.