The Effects of Brominated Flame Retardants on Thyroid Hormone Homeostasis in Human Placenta Tissues and Cell Culture

Polybrominated diphenyl ethers (PBDEs) are a class of brominated flame retardants (BFRs) that have been heavily used in consumer products such as furniture foams, plastics, and textiles since the mid-1970’s. BFRs are added to products in order to meet state flammability standards intended to increase indoor safety in the event of a fire. The three commercial PBDE mixtures, Penta-, Octa-, and DecaBDE, have all been banned in the United States, however, limited use of DecaBDE is still permitted. PBDEs were phased out of production and added to the Stockholm Convention due to concerns over their environmental persistence and toxicity. Human exposure to PBDEs occurs primarily through the inadvertent ingestion of contaminated house dust, as well as though dietary sources. Despite the phase-out and discontinued use of PBDEs, human exposure to this class of chemicals is likely to continue for decades due to the continued use of treated products and existing environmental reservoirs of PBDEs. Extensive research over the years has shown that PBDEs disrupt thyroid hormone (TH) levels and neurodevelopmental endpoints in rodent and fish models. Additionally, there is growing epidemiological evidence linking PBDE exposure in humans to altered TH homeostasis and neurodevelopmental impairments in children. Due to the importance of THs throughout gestation, there is a great need to understand the effects of BFRs on the developing fetus. Specifically, the placenta plays a critical role in the transport, metabolism, and delivery of THs to the fetal compartment during pregnancy and is a likely target for BFR bioaccumulation and endocrine disruption. The central hypothesis of this dissertation research is that BFRs disrupt the activity of TH sulfotransferase (SULT) enzymes, thereby altering TH concentrations in the placenta.

In the first aim of this dissertation research, the concentrations of PBDEs and 2,4,6-TBP were measured in a cohort of 102 placenta tissue samples from an ongoing pregnancy cohort in Durham, NC. Methods were developed for the extraction and analysis of the BFR analytes. It was found that 2,4,6-TBP was significantly correlated with all PBDE analytes, indicating that 2,4,6-TBP may share common product applications with PBDEs or that 2,4,6-TBP is a metabolite of PBDE compounds. Additionally, this was the first study to measure 2,4,6-TBP in human placenta tissues.

In the second aim of this dissertation research, the placenta tissue concentrations of THs, as well as the endogenous activity of deiodinase (DI) and TH SULT enzymes were quantified using the same cohort of 102 placenta tissue samples. Enzyme activity was detected in all samples and this was the first study to measure TH DI and SULT activity in human placenta tissues. Enzyme activities and TH concentrations were compared with BFR concentrations measured in Aim 1. There were few statistically significant associations observed for the combined data, however, upon stratifying the data set based on infant sex, additional significant associations were observed. For example, among males, those with the highest concentrations of BDE-99 in placenta had T3 levels 0.80 times those with the lowest concentration of BDE-99 (95% confidence interval (CI): 0.59, 1.07). Whereas females with the highest concentrations of BDE-99 in placenta had T3 levels 1.50 times those with the lowest concentration of BDE-99 (95% CI: 1.10, 2.04). Additionally, all BFR analyte concentrations were higher in the placenta of males versus females and they were significantly higher for 2,4,6-TBP and BDE-209. 3,3’-T2 SULT activity was significantly higher in female placenta tissues, while type 3 DI activity was significantly higher in male placenta tissues. This research is the first to show sex-specific differences in the bioaccumulation of BFRs in human placenta tissue, as well as differences in TH concentrations and endogenous DI and SULT activity. The underlying mechanisms of these observed sex differences warrant further investigation.

In the third aim of this dissertation research, the effects of BFRs were examined in a human choriocarcinoma placenta cell line, BeWo. Michaelis-Menten parameters and inhibition curves were calculated for 2,4,6-TBP, 3-OH BDE-47, and 6-OH BDE-47. 2,4,6-TBP was shown to be the most potent inhibitor of 3,3’-T2 SULT activity with a calculated IC50 value of 11.6 nM. It was also shown that 2,4,6-TBP and 3-OH BDE-47 exhibit mixed inhibition of 3,3’-T2 sulfation in BeWo cell homogenates. Next, a series of cell culture exposure experiments were performed using 1, 6, 12, and 24 hour exposure durations. Once again, 2,4,6-TBP was shown to be the most potent inhibitor of basal 3,3’-T2 SULT activity by significantly decreasing activity at the high and medium dose (1 M and 0.5 M, respectively) at all measured time points. Interestingly, BDE-99 was also shown to inhibit basal 3,3’-T2 SULT activity in BeWo cells following the 24 hour exposure, despite exhibiting no inhibitory effects in the BeWo cell homogenate experiments. This indicates that BDE-99 must act through a pathway other than direct enzyme inhibition. Following exposures, the TH concentrations in the cell culture growth media and mRNA expression of TH-related genes were also examined. There was no observed effect of BFR treatment on these endpoints. Future work should focus on determining the downstream biological effects of TH SULT disruption in placental cells, as well as the underlying mechanisms of action responsible for reductions in basal TH SULT activity following BFR exposure.

This was one of the first studies to measure BFRs in a cohort of placenta tissue samples from the United States and the first study to measure THs, DI activity, and SULT activity in human placenta tissues. This research provides a novel contribution to our growing understanding of the effects of BFRs on TH homeostasis within the human placenta, and provides further evidence for sex-specific differences within this important organ. Future research should continue to investigate the effects of environmental contaminants on TH homeostasis within the placenta, as this represents the most critical and vulnerable stage of human development.

"On our way": identities and representations of African women seeking asylum in Ireland

This thesis provides the first explicit Postcolonial study of asylum in the Irish context that integrates Black Feminist analyses of intersectional identity with Postcolonial Feminist theories of representation. African women seeking asylum in the Republic of Ireland were key political instruments used by the state to re-draw racial lines. The study examines how, for a group of African women “On their Way” through asylum, identity and representation work hand in hand to force identities, subaltern spaces and bodies to occupy them. Rich biographical data is gathered through mixed art and drama methods over two intensive participatory research projects conducted in a small Irish city. Data analysis critically examines the poetics (practices that signify) and politics (the powers that govern these practices) and affective economies of global and local NGO visual representations, exposing how they consume, fragment, and appropriate African women’s identities and bodies. Though hypervisible, the women themselves “cannot speak”. The women in the study reported feeling “tired” and “used”. Asking “What work are they doing as they do asylum?” the study finds that black female identities and bodies are forced to perform political, cultural, emotional and material labour on their way through this context of Irish asylum. The author argues that Postcolonial Asylum is a performative encounter that re-scripts colonial race/class/gender discourse through a humanitarian alibi to naturalize European/white supremacy, reinscribe patriarchal power and justify racialised incarceration of bodies seeking asylum in the North. This study takes an interdisciplinary approach that centralizes Black and Postcolonial Feminist theory and innovates Participatory Art-Based Action methodology. Black and Postcolonial feminisms can recognize, theorize and replenish black female political and intellectual agency. Participatory Action research, if grounded in Black feminist epistemology and ethics, can allow participants to “speak back” to what is already said about them in spaces of convivial self-representation.

Advanced Placement and American Education: A Foucauldian Analysis of the Advanced Placement Program of the College Board

Advanced Placement is a series of courses and tests designed to determine mastery over introductory college material. It has become part of the American educational system. The changing conception of AP was examined using critical theory to determine what led to a view of continual success. The study utilized David Armstrong’s variation of Michel Foucault’s critical theory to construct an analytical framework. Black and Ubbes’ data gathering techniques and Braun and Clark’s data analysis were utilized as the analytical framework. Data included 1135 documents: 641 journal articles, 421 newspaper articles and 82 government documents. The study revealed three historical ruptures correlated to three themes containing subthemes. The first rupture was the Sputnik launch in 1958. Its correlated theme was AP leading to school reform with subthemes of AP as reform for able students and AP’s gaining of acceptance from secondary schools and higher education. The second rupture was the Nation at Risk report published in 1983. Its correlated theme was AP’s shift in emphasis from the exam to the course with the subthemes of AP as a course, a shift in AP’s target population, using AP courses to promote equity, and AP courses modifying curricula. The passage of the No Child Left Behind Act of 2001 was the third rupture. Its correlated theme was AP as a means to narrow the achievement gap with the subthemes of AP as a college preparatory program and the shifting of AP to an open access program. The themes revealed a perception that progressively integrated the program into American education. The AP program changed emphasis from tests to curriculum, and is seen as the nation’s premier academic program to promote reform and prepare students for college. It has become a major source of income for the College Board. In effect, AP has become an agent of privatization, spurring other private entities into competition for government funding. The change and growth of the program over the past 57 years resulted in a deep integration into American education. As such the program remains an intrinsic part of the system and continues to evolve within American education.

"Per aurem intrat Christus in Mariam". Aproximación iconográfica a la "conceptio per aurem" en la pintura italiana del Trecento desde fuentes patrísticas y teológicas

La tesis mariológica de la conceptio per aurem, según la cual la Virgen María habría concebido a Jesucristo por el oído en el momento de escuchar del ángel el mensaje celestial anunciándole que, sin perder su virginidad, sería madre del Hijo de Dios encarnado, ha merecido hasta ahora muy pocos estudios académicos rigurosamente fundados en fuentes primarias. De hecho, en la literatura especializada son muy escasas las referencias a tal teoría y, cuando algún estudioso la evoca, casi siempre se contenta con aludir a ella, sin aportar pruebas documentales. Sin embargo, tal como lo revelan las nueve pinturas italianas aquí analizadas, esa teoría fue ilustrada mediante sutiles metáforas visuales en muchas obras pictóricas medievales, las cuales se inspiraron en una sólida tradición literaria. Además una pléyade de Padres de la Iglesia y teólogos medievales testimonia, mediante afirmaciones explícitas, que semejante teoría gozó de notable aceptación entre los maestros del pensamiento cristiano. Basándose en numerosos textos patrísticos y teológicos, este artículo intenta dos objetivos esenciales: exponer, ante todo, las distintas formulaciones teóricas propuestas por esos pensadores; y además, tratar de poner en luz los significados dogmáticos que subyacen bajo esa sorprendente tesis.

Ethnicity and the prostate cancer experience: a qualitative metasynthesis

Ethnicity and the prostate cancer experience: a qualitative metasynthesis

Objectives: To summarise black and minority ethnic (BME) patients' and partners
experiences of prostate cancer (PCa) by examining the findings of existing qualitative studies
Methods:
We undertook a systematic metasynthesis of qualitative studies using a modified version of
Noblit and Hare's 'meta-ethnography' approach, with a 2000-2015 search of seven databases.
Results: Thirteen studies of men from US and UK BME groups were included. We explored
constructs with BME-specific features. Healthcare provider relationships, formation of a
spiritual alliance with God (which enhanced the participants’ feeling of empowerment and
ability to cope with the cancer) and living on for others (generally to increase cancer
awareness), often connected to spiritual regrowth, were the three constructs most commonly
reported. A magnified effect from erectile dysfunction was also common. Initially this
affected men’s disclosure to others about their cancer and their sexual problems, but
eventually men responded by shifting their conceptualisations of masculinity to sustain self
and social identities. There was also evidence of inequality resulting from financial
constraints and adversity that necessitated resilience in coping.
Conclusions: The prostate cancer experience of BME men and their partners is affected by a
complex intersection of ethnicity with other factors. Healthcare services should acknowledge
this. If providers recognise the men’s felt masculinities, social identities and spiritual beliefs
and their shifting nature, services could be improved, with community as well as individual
benefits. More studies are needed in diverse ethnic groups

Análise comparativa e teste empírico da validade dos modelos CAPM tradicional e condicional: o caso das ações da Petrobrás

ALVES, Janaína da Silva. Análise comparativa e teste empírico da validade dos modelos CAPM tradicional e condicional: o caso das ações da Petrobrás. Revista Ciências Administrativas, Fotaleza, v. 13, n. 1, p.147-157, ago. 2007.

La funzione di Möbius di un insieme parzialmente ordinato

Nel 1837 il matematico A.F. Möbius definì la funzione aritmetica mu(n) che vale 0 se n è divisibile per il quadrato di un numero primo, (-1)^k se n è il prodotto di k primi distinti e \mu(1)=1. Essa ricopre un ruolo di fondamentale importanza per quanto riguarda la distribuzione dei numeri primi, nonché per la sua duttilità nella risoluzione di diversi problemi di conteggio grazie alla formula di inversione di Möbius, che può essere pensata come un analogo formale del teorema fondamentale del calcolo integrale. Una sorprendente varietà di problemi di calcolo combinatorio si rivelano essere nient'altro che casi particolari di un problema più generale che riguarda la possibilità di invertire una somma fatta sugli elementi di un insieme parzialmente ordinato. L'obiettivo di questo elaborato è quello di illustrare come sia possibile generalizzare il concetto di funzione aritmetica estendendolo a quello di funzione di un'algebra di incidenza. Le algebre di incidenza hanno catturato l'interesse di svariati matematici a partire dagli anni '60 del secolo scorso, e si svilupparono come ambiente naturale nel quale generalizzare la formula di inversione di Mobius. La funzione di Möbius della teoria dei numeri, definita originariamente sull'insieme dei numeri interi positivi ordinato per divisibilità, può quindi essere definita su generici insiemi parzialmente ordinati.

Funzioni BV e disuguaglianze isoperimetriche

All'interno della mia tesi verrà introdotta la teoria delle funzioni in R^{n} a variazione limitata (BV), seguendo le presentazioni di Lawrence C.Evans e Ronald F.Gariepy nel libro Measure Theory and Fine Properties of Functions e di Enrico Giusti nell'opera Minimal Surfaces and Functions of Bounded Variation. Le funzioni BV sono funzioni le cui derivate prime deboli sono misure di Radon, ossia misure di Borel regolari finite sui compatti. In particolare verranno anche analizzati gli insiemi E che hanno perimetro finito, ossia tali che la funzione indicatrice dell’insieme E sia una funzione BV. Nello specifico, nel primo capitolo verranno date le definizioni di funzioni BV e insiemi di perimetro finito, sia in una versione globale che in una locale, verrà enunciato un primo importante teorema per le funzioni BV e verrà analizzata la relazione tra funzioni di Sobolev e funzioni BV. Nel secondo capitolo, invece, verranno analizzate la semicontinuità inferiore, l'approssimazione con funzioni lisce e la compattezza di funzioni BV, mentre nel terzo capitolo verranno elencati alcuni risultati sulle funzioni BV riguardanti la Traccia, l'Estensione e la formula di Coarea. Infine, nel quarto ed ultimo capitolo, verranno studiate le disuguaglianze di Sobolev e Poincaré e le disuguaglianze isoperimetriche per funzioni BV.

Post-translational lysine-acetylation of Ran and its regulation by Sirtuin deacetylases

Through recent advances in high-throughput mass spectrometry it has become evident that post-translational N-(epsilon)-lysine-acetylation is a modification found on thousands of proteins of all cellular compartments and all essential physiological processes. Many aspects in the biology of lysine-acetylation are poorly understood, including its regulation by lysine-acetyltransferases and lysine-deacetylases (KDACs). Here, the role of this modification was investigated for the small GTP-binding protein Ran, which, inter alia, is essential for the regulation of nucleocytoplasmic transport. To this end, site-specifically acetylated Ran was produced in E. coli by genetic code expansion. For five previously identified sites, Ran acetylation was tested regarding its impact on the intrinsic GTP hydrolysis rate, the assembly of export complexes (modeled in vitro with the export receptor CRM1 and the export substrate Spn1) and the interaction of Ran with its GTPase activation protein RanGAP and RanBP1. Overall, mild effects of Ran acetylation were observed for intrinsic and RanGAP-stimulated GTP hydrolysis rates. The interaction of active Ran with RanBP1 was negatively influenced by Ran acetylation at K159. Moreover, CRM1 bound to Ran acetylated at K37, K99 or K159 interacted more strongly with Spn1. Thus, lysine-acetylation interferes with essential aspects of Ran function. An in vitro screen was performed to identify potential Ran KDACs. The NAD+-dependent KDACs of the Sirtuin class showed activity towards two acetylation sites of Ran, K37 and K71. The specificity of Sirtuins was further analyzed based on an additional Ran acetylation site, K38. Since deacetylation of RanAcK38 was much slower compared to RanAcK37, di-acetylated RanAcK37/38 was tested next. The deacetylation rate of di-acetylated Ran was comparable to that of RanAcK37. Deacetylation experiments under single turnover conditions revealed that deacetylation occurs first at the K38 site in the di-acetylated RanAcK37/38 background. The ability of Sirtuins to deacetylate two adjacent AcKs was further investigated based on two proteins, which had previously been found to be di-acetylated and targeted by Sirtuins, namely the tumor suppressor protein p53 and phosphoenolpyruvate carboxykinase 1 (PEPCK1). p53 was readily deacetylated at two di-acetylation sites (K372/372 and K381/382), whereas PEPCK1 was not deacetylated in vitro. Taken together, these results have important implications for the substrate specificity of Sirtuins.

Análise comparativa e teste empírico da validade dos modelos CAPM tradicional e condicional: o caso das ações da Petrobrás

ALVES, Janaína da Silva. Análise comparativa e teste empírico da validade dos modelos CAPM tradicional e condicional: o caso das ações da Petrobrás. Revista Ciências Administrativas, Fotaleza, v. 13, n. 1, p.147-157, ago. 2007.

The analysis and detection of new psychoactive substances in biological matrices

New psychoactive substances (NPSs) have appeared on the recreational drug market at an unprecedented rate in recent years. Many are not new drugs but failed products of the pharmaceutical industry. The speed and variety of drugs entering the market poses a new complex challenge for the forensic toxicology community. The detection of these substances in biological matrices can be difficult as the exact compounds of interest may not be known. Many NPS are sold under the same brand name and therefore users themselves may not know what substances they have ingested. The majority of analytical methods for the detection of NPSs tend to focus on a specific class of compounds rather than a wide variety. In response to this, a robust and sensitive method was developed for the analysis of various NPS by solid phase extraction (SPE) with gas chromatography mass spectrometry (GCMS). Sample preparation and derivatisation were optimised testing a range of SPE cartridges and derivatising agents, as well as derivatisation incubation time and temperature. The final gas chromatography mass spectrometry method was validated in accordance with SWGTOX 2013 guidelines over a wide concentration range for both blood and urine for 23 and 25 analytes respectively. This included the validation of 8 NBOMe compounds in blood and 10 NBOMe compounds in urine. This GC-MS method was then applied to 8 authentic samples with concentrations compared to those originally identified by NMS laboratories. The rapid influx of NPSs has resulted in the re-analysis of samples and thus, the stability of these substances is crucial information. The stability of mephedrone was investigated, examining the effect that storage temperatures and preservatives had on analyte stability daily for 1 week and then weekly for 10 weeks. Several laboratories identified NPSs use through the cross-reactivity of these substances with existing screening protocols such as ELISA. The application of Immunalysis ketamine, methamphetamine and amphetamine ELISA kits for the detection of NPS was evaluated. The aim of this work was to determine if any cross-reactivity from NPS substances was observed, and to determine whether these existing kits would identify NPS use within biological samples. The cross- reactivity of methoxetamine, 3-MeO-PCE and 3-MeO-PCP for different commercially point of care test (POCT) was also assessed for urine. One of the newest groups of compounds to appear on the NPS market is the NBOMe series. These drugs pose a serious threat to public health due to their high potency, with fatalities already reported in the literature. These compounds are falsely marketed as LSD which increases the chance of adverse effects due to the potency differences between these 2 substances. A liquid chromatography tandem mass spectrometry (LC-MS/MS) method was validated in accordance with SWGTOX 2013 guidelines for the detection for 25B, 25C and 25I-NBOMe in urine and hair. Long-Evans rats were administered 25B-, 25C- and 25I-NBOMe at doses ranging from 30-300 µg/kg over a period of 10 days. Tail flick tests were then carried out on the rats in order to determine whether any analgesic effects were observed as a result of dosing. Rats were also shaved prior to their first dose and reshaved after the 10-day period. Hair was separated by colour (black and white) and analysed using the validated LC-MS/MS method, assessing the impact hair colour has on the incorporation of these drugs. Urine was collected from the rats, analysed using the validated LC-MS/MS method and screened for potential metabolites using both LC-MS/MS and quadrupole time of flight (QToF) instrumentation.

[Map of proposed extensions of Vermont and New Jersey avenues N.W. between Florida and Georgia avenues, Washington D.C.].

Cadastral map showing lots and buildings in path of proposed avenue extensions.

[Map of part of S.E. Washington D.C. now occupied by the Navy Yard and adjacent squares].

Covers area bounded by Anacostia River waterfront, 10th, L, and 5th streets S.E.

Empirical validation of CAPM

This paper pretends to show empirical evidence of the CAPM model of Sharpe-Lintner (1964) for Colombia from 2003 to 2010, whose validation is carried out using the method of Black, Jensen and Scholes (1972) but introducing certain methodological econometric type changes associated to the requirements imposed by the used sample -- Specifically, we found no empirical evidence to reject the CAPM for the Colombian economyin the period under analysis