An analysis of state crime: negotiating multiple insecurities around the U.S. Mexico Border

This research looks into forms of state crime taking place around the U.S.-Mexico border. On the Mexican side of the border violent corruption and criminal activities stemming from state actors complicity with drug trafficking organisations has produced widespread violence and human casualty while forcing many to cross the border legally or illegally in fear for their lives. Upon their arrival on the U.S. side of the border, these individuals are treated as criminal suspects. They are held in immigration detention facilities, interrogated and categorised as inadmissible ‘economic migrants’ or ‘drug offenders’ only to be denied asylum status and deported to dangerous and violent zones in Mexico. These individuals have been persecuted and victimised by the state during the 2007-2012 counter narcotic operations on one side of the border while criminalised and punished by a categorizing anti-immigration regime on the other side of the border. This thesis examines this border crisis as injurious actions against border residents have been executed by the states under legal and illegal formats in violation of criminal law and human rights conventions. The ethnographic research uses data to develop a nuanced understanding of individuals’ experiences of state victimisation on both sides of the border. In contributing to state crime scholarship it presents a multidimensional theoretical lens by using organised crime theoretical models and critical criminology concepts to explain the role of the state in producing multiple insecurities that exclude citizens and non-citizens through criminalisation processes.

Spliceosomal small nuclear ribonucleoprotein particle trafficking and maturation in the nuclear compartment

We show for the first time that upon injection into the cytoplasm of the oocyte, fluorescein-labeled spliceosomal snRNAs, in the context of functional snRNPs, are targeted to elongating pre-mRNAs. This finding presents us with a novel assay with which to dissect the mechanism by which snRNPs are targeted to nascent pre-mRNA transcripts. Two critical advantages offered by this system are immediately evident. First, it allows us to investigate the mechanisms employed to recruit snRNPs as it actually transpires within the realm of the cell nucleus. Second, it allows a genome-wide analysis of snRNP recruitment to nascent transcripts, and, hence, the conclusions drawn from these studies do not depend on the sequence of any particular promoter or pre-mRNA. Indeed, it is with this assay that we have stumbled upon a most unanticipated discovery: Contrary to the current paradigm, the co-transcriptional recruitment of splicing snRNPs to nascent transcripts is not contingent on their role in splicing in vivo. Based on these and other data, we have constructed a two-step recruitment-loading model wherein snRNPs are first recruited to pre-mRNA transcripts and only then loaded directly onto cis-acting sequences on nascent pre-mRNA. While conducting studies on snRNP trafficking, a new discovery was made. We found that the lampbrush chromosomes could be visualized by light microscopy in vivo, and that these chromosomes have an architecture that is identical with those in formaldehyde treated nuclear spread preparations. Importantly, we now have the first system with which we can examine the dynamic interactions of macromolecules with specific RNA polymerase II transcriptional units in the live nucleus.

Regulatory crosstalk by protein kinases on CFTR trafficking and activity

Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) is a member of the ATP binding cassette (ABC) transporter superfamily that functions as a cAMP-activated chloride ion channel in fluid-transporting epithelia. There is abundant evidence that CFTR activity (i.e., channel opening and closing) is regulated by protein kinases and phosphatases via phosphorylation and dephosphorylation. Here, we review recent evidence for the role of protein kinases in regulation of CFTR delivery to and retention in the plasma membrane. We review this information in a broader context of regulation of other transporters by protein kinases because the overall functional output of transporters involves the integrated control of both their number at the plasma membrane and their specific activity. While many details of the regulation of intracellular distribution of CFTR and other transporters remain to be elucidated, we hope that this review will motivate research providing new insights into how protein kinases control membrane transport to impact health and disease.

La protección a las víctimas de trata de personas en Brasil

La trata de personas es una acción ilegal presente en todomundo, incluso en Brasil. El Protocolo de las Naciones Unidascontra la Trata de Personas fue ratificado por Brasil en 2004. Apesar de esto, aún la evolución normativa del país no ha sidosignificativa. Este artículo busca analizar la protección normativaexistente y futura a las víctimas, especialmente extranjeras, detrata de personas en Brasil. Así, inicialmente se presenta lo que secomprende como trata de personas, con base en el Protocolo delas Naciones Unidas para prevenir, reprimir y sancionar la trata depersonas, especialmente mujeres y niños, y son apuntadas algunasconsideraciones sobre la realidad de la trata de personas en elmundo y en Brasil. En la secuencia son presentadas las medidasde protección a las víctimas, y previstas en el Protocolo de lasNaciones Unidas. Por último, son enumeradas y analizadas lasacciones normativas existentes y los proyectos de ley que puedencambiar esta realidad. La conclusión del estudio indica que Brasilaún tiene una débil protección para las víctimas extranjeras detrata de personas, pero hay una perspectiva optimista de mejoríacon los proyectos de ley que tramitan en el Congreso Nacional.

Tissue and cell's trafficking: a byproduct of transplant activity's trash. The future is now. A Public Health problem

Transplantation is one of the most beautiful achievements for humanity in the last century and became the last hope to many patients. As other beautiful achievements, it has been used by criminals. The future of transplantation will be focused on tissue and cells transplantation. Trafficking of human beings to organ removal and trafficking of human organs are an early stage of trafficking on tissues and cells comparable with slaves trafficking in the 17th and 18th century. As 400 years ago, the motive for the crime is development, economy and profit. Transplant surgery is the modern “cotton gin” to this new commerce. Poverty exploitation, unprotected people, are always the victims. Even so, there are some differences since then. The paying buyers are the patients themselves and the “cotton” transplanted is not so harmless. Unsafe tissues and cells inappropriately collected and allocated can be so dangerous to the recipient and his family, that the dreamed transplant/implant becomes a nightmare. Beyond the trafficking crime, there is a most dangerous associated crime that is the crime of spreading dangerous infectious diseases.

More Than A Movement: Unpacking Contemporary Anti-Human Trafficking Efforts in Washington, D.C.

Trafficking in persons has attracted seemingly boundless attention over the last two decades and the work aimed at fighting it is best understood when this cause is contextualized against the backdrop of other social forces—economic, social, and cultural—shaping contemporary nonprofit activities. This project argues that the paid and volunteer labor that takes place in metro Washington, D.C., to combat trafficking in persons can be understood as both a movement and an industry. In addition to arguing that anti-trafficking work is part of a nonprofit industrial complex that situates activist and advocacy work firmly inside state and economic institutions, this project is concerned with the ways in which trafficking work and workers conduct their business collectively. As an organizational study, it identifies the key players in the D.C. region focused on this issue and traces their interactions, collaborations, and cooperation. Significantly, this project suggests that despite variations in objectives, methods, priorities, and characterizations of trafficking, thirty organizations in metro D.C. working on this issue “get along” because they are bound by the benign common goal of raising awareness. Awareness, in this context, is best understood as both a cultural anchor facilitating cohesion and as a social currency allowing groups to opt into joint efforts. The dissertation concludes that organizations centralize awareness in their collective activities over more drastic priorities around which consensus would need to be gained. This is a lost opportunity for making sense of the ways that individual bodies—men, women, and children—experience not just trafficking, but the world around them.

Estudo sócio-ecológico sobre tráfico de seres humanos para fins de exploração laboral na região do Alentejo : análise espacial e estatística

O presente relatório, inserido no Mestrado em Gestão do Território, Área de Especialização em Deteção Remota e Sistemas de Informação Geográfica, lecionado pelo Departamento de Geografia e Planeamento Regional da Faculdade de Ciências Sociais e Humanas da Universidade Nova de Lisboa, pretende descrever o trabalho desenvolvido pelo mestrando enquanto estagiário no Observatório do Tráfico de Seres Humanos (OTSH). O relatório está estruturado em três capítulos distintos. No primeiro capítulo é realizada uma abordagem teórica sobre o Tráfico de Seres Humanos e a distinção entre o mesmo com o Auxílio à Imigração Ilegal. Neste, é também feita uma pequena referência à problemática dos novos fluxos de refugiados/migrantes que, no momento da realização do mesmo, constituem uma questão bastante complexa sobretudo ao nível europeu. No segundo capítulo é realizada uma caracterização da área de estudo, assim como a descrição dos dados utilizados e a metodologia aplicada no mesmo. No terceiro capítulo são apresentados os resultados finais do estudo e a cartografia de síntese que sustenta os mesmos. Para a realização deste estudo recorreu-se a uma análise multicritério em SIG para prever a localização de áreas de maior suscetibilidade de ocorrência de novos casos relativos ao crime do tráfico de seres humanos para exploração laboral na agricultura, na região do Alentejo (distritos de Beja, Évora e Portalegre), através do recurso a dados estatísticos disponibilizados tanto pelo OTSH, como por outras entidades. A metodologia apresentada integra um SIG baseado num modelo raster com o Analytical Hierarchy Process (AHP). Através da realização deste estudo, a importância dos SIG como ferramenta no auxílio ao processo de tomada de decisão, pôde ser testada, conjuntamente com o processo metodológico AHP, através dos resultados apresentados. Com um possível desenvolvimento deste modelo analítico, pretende-se que o mesmo seja adaptável a outras regiões e em última instância, outros tipos de exploração e/ou tráfico.

New insights into plant salt acclimation: the roles of vesicle trafficking and reactive oxygen species signalling in mitochondria and the endomembrane system

In this study, we investigated the cellular and molecular mechanisms that regulate salt acclimation. The main objective was to obtain new insights into the molecular mechanisms that control salt acclimation. Therefore, we carried out a multidisciplinary study using proteomic, transcriptomic, subcellular and physiological techniques. We obtained a Nicotiana tabacum BY-2 cell line acclimated to be grown at 258 mM NaCl as a model for this study. The proteomic and transcriptomic data indicate that the molecular response to stress (chaperones, defence proteins, etc.) is highly induced in these salt-acclimated cells. The subcellular results show that salt induces sodium compartmentalization in the cell vacuoles and seems to be mediated by vesicle trafficking in tobacco salt-acclimated cells. Our results demonstrate that abscisic acid (ABA) and proline metabolism are crucial in the cellular signalling of salt acclimation, probably regulating reactive oxygen species (ROS) production in the mitochondria. ROS may act as a retrograde signal, regulating the cell response. The network of endoplasmic reticulum and Golgi apparatus is highly altered in salt-acclimated cells. The molecular and subcellular analysis suggests that the unfolded protein response is induced in salt-acclimated cells. Finally, we propose that this mechanism may mediate cell death in salt-acclimated cells.

Intracellular signaling and trafficking in cancer: Role of Rab5-GTPases in migration and invasion of breast cancer cells

Metastasis is characterized pathologically by uncontrolled cell invasion, proliferation, migration and angiogenesis. Steroid hormones, such as estrogen, and growth factors, which include insulin growth factor I/II (IGF-1/IGF-2) therapy has been associated with most if not all of the features of metastasis. It has been determined that IGF-1 increases cell survival of cancer cells and potentiate the effect of E2 and other ligand growth factors on breast cancer cells. However not much information is available that comprehensively expounds on the roles of insulin growth factor receptor (IGFR) and Rab GTPases may play in breast cancer. The latter, Rab GTPases, are small signaling molecules and critical in the regulation of many cellular processes including cell migration, growth via the endocytic pathway. This research involves the role of Rab GTPases, specifically Rab5 and its guanine exchange factors (GEFs), in the promotion of cancer cell migration and invasion. Two important questions abound: Are IGFR stimulation and downstream effect involved the endocytic pathway in carcinogenesis? What role does Rab5 play in cell migration and invasion of cancer cells? The hypothesis is that growth factor signaling is dependent on Rab5 activity in mediating the aggressiveness of cancer cells. The goal is to demonstrate that IGF-1 signaling is dependent on Rab5 function in breast cancer progression. Here, the results thus far, have shown that while activation of Rab5 may mediate increased cell proliferation, migration and invasion in breast cancer cells, the Rab5 GEF, RIN1 interacts with the IGFR thereby facilitating migration and invasion activities in breast cells. Furthermore, endocytosis of the IGFR in breast cancer cells seems to be caveolin dependent as the data has shown. This taken together, the data shows that IGF-1 signaling in breast cancer cells relies on IGF-1R phosphorylation, caveolae internalization and sequestration to the early endosome RIN1 function and Rab5 activation.^

Introduction of African Swine Fever into the European Union through Illegal Importation of Pork and Pork Products

Transboundary animal diseases can have very severe socio-economic impacts when introduced into new regions. The history of disease incursions into the European Union suggests that initial outbreaks were often initiated by illegal importation of meat and derived products. The European Union would benefit from decision-support tools to evaluate the risk of disease introduction caused by illegal imports in order to inform its surveillance strategy. However, due to the difficulty in quantifying illegal movements of animal products, very few studies of this type have been conducted. Using African swine fever as an example, this work presents a novel risk assessment framework for disease introduction into the European Union through illegal importation of meat and products. It uses a semi-quantitative approach based on factors that likely influence the likelihood of release of contaminated smuggled meat and products, and subsequent exposure of the susceptible population. The results suggest that the European Union is at non-negligible risk of African swine fever.

Functional analysis of Arabidopsis thaliana lesion mimic mutant cfs1 in ESCRT complex-related protein trafficking

The Endosomal Sorting Complex Required for Transport (ESCRT)-complex is composed of four complexes, ESCRT-0-III. They sequentially act on a late endosome to sort mono-ubiquitinated transmembrane proteins into the intralumenal vesicle, forming of a multivesicular body(MVB) that is delivered to vacuole for degradation. In Arabidopsis thaliana, the loss of an ESCRT-I component, elch displays a cytokinesis defect; while a dominant negative expression of an ESCRT-III component results in cell death due to vacuolar loss. In this work, the function of a plant-specific ELCH-interactor, CELL DEATH RELATED FYVE/SYLF DOMAIN CONTAINING 1 (CFS1) and its influences on the ESCRT-complex function are investigated. CFS1 is a phosphatidylinositol-3-phosphate- and actin-binding protein. The cfs1 mutants mimic lesions in the first eldest leaf that propagate to the next eldest one. Genetic analyses have demonstrated that cell death in cfs1 does not require a functional ESCRT-I component; nevertheless, the loss of CFS1 alleviates elchcytokinesis defect, suggesting its influence on the ESCRT-I function. Further analyses reveal that cfs1 accumulates autophagosomes throughout its lifespan due to a decrease in autophagosome degradation, suggesting that as the plant ages, the cumulated autophagosomes falsely trigger effectors-triggered immunity that executes cell death in cfs1. As the ESCRT-complex has been demonstrated to be involved in the delivery of autophagosomes to vacuole and CFS1 homolog, CFS2 reportedly interacts with ATG8, it can be postulated from the results of this work that CFS1 alone or together with CFS2 function in sequestering mature autophagosomes onto MVBs. At the MVBs, the ESCRT-complex then mediates the fusion of autophagosome and MVB for subsequent delivery to vacuole.

La migración infantil indocumentada de Centroamérica, un desafío a la reformulación de la política migratoria de Estados Unidos. Estudio de caso de Honduras (2010-2014)

El siguiente estudio de caso analiza la influencia de la llegada de la migración infantil indocumentada centroamericana en la reformulación de la política migratoria de Estados Unidos en el período 2010-2014. Enfocándose en el caso de Honduras para dar a conocer y analizar las causas que crean las dinámicas migratorias por parte de los menores, este trabajo analiza como la llegada de estas poblaciones genera ciertos efectos en el proceso de toma de decisión de las políticas internas de los Estados Unidos. Por un lado, para resaltar las características del fenómeno migratorio, se utilizan las teorías de redes sociales y la teoría push and pull. Por otro, mediante los conceptos de Sensibilidad y Vulnerabilidad expuestos en la teoría de la Interdependencia Compleja de las Relaciones Internacionales, como también el concepto de Seguridad Societal propuesto por Barry Buzan se estudia el nivel de influencia del fenómeno infantil en el gobierno norteamericano.

Francisco Pizarro' s banner of arms: an analytical work contributing to latin America's history

The study of textiles is an open area of scientific research, which for its variety of material components and physical chemical diversity of conditions, makes a field of interest for scientific studies in the cultural heritage field. Archaeological/historical textiles offer the possibility to carry out studies on organic materials such as fibers, adhesion elements, dyes, paper, etc., as well as on inorganic compounds for instance metals, alloys, precious stones and other added ornamentation. That variety of composition, allow to use a combination of analytical techniques to solve the questions coming from the object in an archaeometric research. One kind of textile object that provides a valuable cultural information because of its linguistic representation employed by its carrier societies, are the flags/banners/emblems, objects made with a nonverbal communication purpose. As long as depending on the use and/or purpose of each object, varies both the materials/techniques used in its production and its iconography (style, color, emblem, shape), its study gives the possibility to extract information through their materials and manufacturing techniques about a temporal-spatial frame, a particular event or a specific character. The flags/banners have been used since the eleventh century as representative objects of power, hierarchy, social or military organization, or as communicative media. The use of these objects has been spread throughout the world, possibly due to its easy interpretation and/or appropriation by different societies, making it part of their own culture. The flags as symbols of territorial control, using emblems that represent a family, order or army, were introduced to the New World (America) with the arrival of the European conquerors at the end of the fifteenth century. Flags/banners representing the Royal dominion over conquered territories, the Catholic Church and conquistadors’ armies were the first to arrive. One of those flags that have endured over time, that have an invaluable cultural meaning for both American and Iberian societies, is the so-called Francisco Pizarro’s Banner of Arms. It is a textile object with metal threads decoration over a Royal emblem. According to historical sources, this object was used by Francisco Pizarro in 1532 on the conquest process of Peru, after received the permission by King Charles V to on behalf of him, to conquer the lands of the New World today known as Peru. After Pizarro’s control of the Inca territory, it is believed that Pizarro left his banner on top of the Inca’s Sun’s Temple as symbol of his rule. Centuries later, in the America libertarian campaigns, General Sucre, military at charge of the independence army in Peru, reports have found what he considered the Pizarro’s Banner, sending it to Bogotá as a symbol of victory, being kept since that time until today by the National Museum of Colombia. Due to historical discrepancies in the different movements of the so-called Pizarro’s Banner of Arms, its real meaning has been under discussion and because of the passage of time its physical condition has suffer deterioration. That is because its scientific study is now an interesting case study to respond to both historical and conservation questions of it. Through a collaboration with the National Museum of Colombia, a set of 25 samples of so-called Pizarro’s Banner of Arms were collected, covering the various components and areas from the object of study. These samples were subjected to analytical studies for physical and chemical characterization. Microscopic observation, VSEM-EDS analysis, Raman spectroscopy, chromatographic analysis (HPLC-MS, GCMS) and radiocarbon dating were done. Similarly, was sought through a direct in situ physical inspection to the object and through a research into historical sources, adequate information to solve the object’s problems. Results obtained allowed to identify as silk the textile used in the elaboration of the Banner’s fabric, as well as the use of natural dyes for dyeing the fibers used on the emblem: use of cochineal and brazil wood as a source of red, luteolin plant-based for yellow color, indigotine plant-based for blue, and a mixture of yellow and blue dyes for green were identified. Similarly, the use of animal glue in the manufacturing process and the use of rag paper was evident. The metal threads study from the Banner give a confirmation to a silver core wire gilded with a thin gold sheet, being flattened and entwined with silk threads for their use. Finally, using the radiocarbon results, it was possible to postulate with huge accuracy that the Banner date manufacture was between the XV-XVI century and subject to restoration processes with addition of textiles in modern times. Together with, was evident that the state of degradation of the fabric is due to natural degradation in the silk fibers, having that its color has faded and its mechanical properties decreased, leading to loss of rigidity and disappearance of the physical structure. Similarly, it was clear the original colors of the emblem and highlight problems of detachment of paper due to crystallization of the adhesive. In the same way, was found that the metal threads suffer corrosion by sulfur and detachment of its crystals. Finally, combining the analytical results and the historical sources data found from the so-called Francisco Pizarro’s Banner of Arms, allows to postulate that its manufacture process was done in Europe employing precious materials to obtain a long-life object with a deep message for its viewers. Also, the data obtained helps to support the possible idea that the object was employed by Francisco Pizarro in the Peru conquest process. However, by the symbols present in the object, its elaboration date and materials, this object its clearly unique in its kind, and the most important, by its linguistic message, does not represent to Francisco Pizarro or his army, meanwhile, represents the Spanish crown. Therefore, instead to be labeled as Francisco Pizarro’s Banner of Arms, it should be called the Colonial Royal Banner of Charles V in the New World; RESUMEN: El estudio de textiles es un área abierta de investigación científica, la cual por su variedad de componentes materiales y la diversidad de condiciones físico-químicas presentes en estos objetos, lo hace un campo de interés para estudios científicos en el patrimonio cultural. Los textiles arqueológicos/históricos brindan la posibilidad de realizar estudios en materiales orgánicos como fibras, elementos de adhesión, tinturas, papel, etc., e inorgánicos como metales, aleaciones, piedras preciosas y demás materiales decorativos añadidos. Por su variedad de composición, es posible emplear diversas técnicas analíticas para resolver aquellas preguntas propias del objeto en una investigación arqueométrica. Uno de los objetos textiles que brinda gran información cultural debido a su representación lingüística empleada por las sociedades portadoras, son las banderas/estandartes/emblemas. Donde varía dependiendo de su uso y/o propósito, los materiales empleados en su elaboración, al igual que su iconografía (estilo, color, emblema, forma). El estudio de estos objetos construidos con un propósito de comunicación no verbal, da la posibilidad de extraer información a través de sus materiales y técnicas de elaboración sobre un rango temporal-espacial, un evento determinado en la historia o incluso a un personaje en específico. Las banderas han sido empleadas desde el siglo XI como objetos representativos de poder, jerarquía, organización social o militar, o como medio de comunicación. El uso de estos objetos se ha extendido a lo largo del mundo posiblemente debido a su fácil interpretación y/o apropiación por distintas sociedades, haciéndolo parte de su cultura. Las banderas como símbolos de control territorial, empleando símbolos que representan a una familia, orden o armada fueron introducidas a el Nuevo Mundo (América) con la llegada de los conquistadores europeos al final del siglo XV. Las banderas/estandartes que representaban el dominio Real sobre territorios dominados, la iglesia católica y las banderas de ejércitos y/o conquistadores fueron las primeras en llegar al nuevo mundo. Una de aquellas banderas que ha soportado el paso del tiempo, teniendo un gran valor cultural tanto para las sociedades americanas como para las ibéricas, es el denominado Estandarte de armas de Francisco Pizarro. Siendo un objeto textil con decoración en hilos metálicos sobre un emblema Real. De acuerdo a fuentes históricas, este objeto fue usado por Francisco Pizarro en 1532 en el proceso de conquista del Perú, quien recibe por parte del Rey Carlos V el poder para que, en su nombre, Pizarro pueda conquistar las tierras del nuevo mundo hoy conocidas como Perú. Luego del dominio de Pizarro sobre el territorio Inca, se cree que Pizarro dejó su estandarte en la cima del templo Inca del sol como símbolo de su control. Siglos más tarde, en las campañas libertarias de América, el General Sucre, militar encargado de la armada independentista en Perú, reporta haber encontrado lo que él considera como el estandarte de Pizarro, enviándolo a Bogotá como muestra de victoria, siendo custodiada desde ese momento por el Museo Nacional de Colombia hasta la actualidad. Debido a discrepancias históricas, el verdadero significado del llamado estandarte de Pizarro ha sido objeto de discusión y debido del pasar del tiempo su estado de conservación se ha deteriorado. Dejando de este modo, un caso de estudio interesante para que por medio de estudios científicos al objeto se pueda dar respuesta a preguntas tanto históricas como de conservación del mismo. De este modo, por medio de una colaboración con el Museo Nacional de Colombia, se obtuvo un juego de 25 muestras del llamado Estandarte de armas de Francisco Pizarro, abarcando los diferentes componentes y áreas del objeto de estudio. Dichas muestras fueron sometidas a estudios analíticos para su caracterización físico-química. Análisis de observación al microscopio, análisis VSEM-EDS, espectroscopia Raman, análisis cromatográficos (HPLC-MS, GC-MS) y datación por radiocarbono catorce fueron realizados. Del mismo modo, por medio de una inspección física al objeto in situ y una profunda investigación en fuentes históricas del mismo, se buscó la información adecuada para resolver sus problemáticas. Los resultados obtenidos permitieron identificar como seda el textil empleado en la elaboración del estandarte, así como el uso de colorantes naturales para teñir las fibras en el emblema: uso de cochinilla y palo de Brasil como fuente del color rojo, plantas a base de luteolin para el color amarillo, plantas a base de indigotina para el color azul y mezcla de colorantes amarillos y azules para el color verde fueron identificadas. Del mismo modo se evidencio el uso de adhesivos animales y el uso de papel de trapos en el proceso de manufactura. El estudio de los hilos metálicos, permitió evidenciar el uso de alambres con núcleos de plata con un fino recubrimiento de oro en su exterior, siendo aplanados y entrelazados con hilos de seda para su uso. Finalmente usando la datación por radiocarbono, fue posible conocer con alta precisión que el estandarte fue elaborado entre los siglos XV-XVI y sufrió procesos de restauración con añadidura de textiles en tiempos modernos. Junto a lo anterior, es posible postular que el estado de degradación de la tela es debido a degradación natural en las fibras de seda, teniendo así que su color se ha desvanecido y sus propiedades mecánicas disminuidas, conllevando a perdida de rigidez y desaparición de la estructura. Del mismo modo se pudo conocer los colores originales del emblema y evidenciar problemas de desprendimiento del papel debido a cristalización del adhesivo. Asimismo, se comprobó que los hilos metálicos presentan corrosión por azufre y desprendimiento de sus cristales. Finalmente, combinando los resultados analíticos y la información de fuentes históricas encontradas del llamado Estandarte de armas de Francisco Pizarro, se puede postular que su elaboración fue realizada en Europa, usando materiales preciosos para obtener un objeto de larga vida con un profundo mensaje para sus observadores. También, los datos obtenidos ayudan a dar soporte la posible idea de que este objeto fue usado por Francisco Pizarro en el proceso de conquista del Perú. Sin embargo, debido a los símbolos presentes en el objeto, fecha y materiales de elaboración, este objeto es claramente único en su tipo, y lo más importante, por su mensaje lingüístico, este no representa a Francisco Pizarro o su armada, al contrario, representa a la Corona de España. Por ende, en vez de denominarse como Estandarte de armas de Francisco Pizarro, este objeto debería nombrarse como el Estandarte Real de la Colonia de Carlos V en el Nuevo Mundo.

Membrane associated transporter protein gene (SLC45A2) and the genetic basis of normal human pigmentation variation

This work is concerned with the genetic basis of normal human pigmentation variation. Specifically, the role of polymorphisms within the solute carrier family 45 member 2 (SLC45A2 or membrane associated transporter protein; MATP) gene were investigated with respect to variation in hair, skin and eye colour ― both between and within populations. SLC45A2 is an important regulator of melanin production and mutations in the gene underly the most recently identified form of oculocutaneous albinism. There is evidence to suggest that non-synonymous polymorphisms in SLC45A2 are associated with normal pigmentation variation between populations. Therefore, the underlying hypothesis of this thesis is that polymorphisms in SLC45A2 will alter the function or regulation of the protein, thereby altering the important role it plays in melanogenesis and providing a mechanism for normal pigmentation variation. In order to investigate the role that SLC45A2 polymorphisms play in human pigmentation variation, a DNA database was established which collected pigmentation phenotypic information and blood samples of more than 700 individuals. This database was used as the foundation for two association studies outlined in this thesis, the first of which involved genotyping two previously-described non-synonymous polymorphisms, p.Glu272Lys and p.Phe374Leu, in four different population groups. For both polymorphisms, allele frequencies were significantly different between population groups and the 272Lys and 374Leu alleles were strongly associated with black hair, brown eyes and olive skin colour in Caucasians. This was the first report to show that SLC45A2 polymorphisms were associated with normal human intra-population pigmentation variation. The second association study involved genotyping several SLC45A2 promoter polymorphisms to determine if they also played a role in pigmentation variation. Firstly, the transcription start site (TSS), and hence putative proximal promoter region, was identified using 5' RNA ligase mediated rapid amplification of cDNA ends (RLM-RACE). Two alternate TSSs were identified and the putative promoter region was screened for novel polymorphisms using denaturing high performance liquid chromatography (dHPLC). A novel duplication (c.–1176_–1174dupAAT) was identified along with other previously described single nucleotide polymorphisms (c.–1721C>G and c.–1169G>A). Strong linkage disequilibrium ensured that all three polymorphisms were associated with skin colour such that the –1721G, +dup and –1169A alleles were associated with olive skin in Caucasians. No linkage disequilibrium was observed between the promoter and coding region polymorphisms, suggesting independent effects. The association analyses were complemented with functional data, showing that the –1721G, +dup and –1169A alleles significantly decreased SLC45A2 transcriptional activity. Based on in silico bioinformatic analysis that showed these alleles remove a microphthalmia-associated transcription factor (MITF) binding site, and that MITF is a known regulator of SLC45A2 (Baxter and Pavan, 2002; Du and Fisher, 2002), it was postulated that SLC45A2 promoter polymorphisms could contribute to the regulation of pigmentation by altering MITF binding affinity. Further characterisation of the SLC45A2 promoter was carried out using luciferase reporter assays to determine the transcriptional activity of different regions of the promoter. Five constructs were designed of increasing length and their promoter activity evaluated. Constitutive promoter activity was observed within the first ~200 bp and promoter activity increased as the construct size increased. The functional impact of the –1721G, +dup and –1169A alleles, which removed a MITF consensus binding site, were assessed using electrophoretic mobility shift assays (EMSA) and expression analysis of genotyped melanoblast and melanocyte cell lines. EMSA results confirmed that the promoter polymorphisms affected DNA-protein binding. Interestingly, however, the protein/s involved were not MITF, or at least MITF was not the protein directly binding to the DNA. In an effort to more thoroughly characterise the functional consequences of SLC45A2 promoter polymorphisms, the mRNA expression levels of SLC45A2 and MITF were determined in melanocyte/melanoblast cell lines. Based on SLC45A2’s role in processing and trafficking TYRP1 from the trans-Golgi network to stage 2 melanosmes, the mRNA expression of TYRP1 was also investigated. Expression results suggested a coordinated expression of pigmentation genes. This thesis has substantially contributed to the field of pigmentation by showing that SLC45A2 polymorphisms not only show allele frequency differences between population groups, but also contribute to normal pigmentation variation within a Caucasian population. In addition, promoter polymorphisms have been shown to have functional consequences for SLC45A2 transcription and the expression of other pigmentation genes. Combined, the data presented in this work supports the notion that SLC45A2 is an important contributor to normal pigmentation variation and should be the target of further research to elucidate its role in determining pigmentation phenotypes. Understanding SLC45A2’s function may lead to the development of therapeutic interventions for oculocutaneous albinism and other disorders of pigmentation. It may also help in our understanding of skin cancer susceptibility and evolutionary adaptation to different UV environments, and contribute to the forensic application of pigmentation phenotype prediction.