Altered expression of the ToxR-regulated porins OmpU and OmpT diminishes Vibrio cholerae bile resistance, virulence factor expression, and intestinal colonization

The transmembrane transcriptional activators ToxR and TcpP modulate expression of Vibrio cholerae virulence factors by exerting control over toxT, which encodes the cytoplasmic transcriptional activator of the ctx, tcp, and acf virulence genes. However, ToxR, independently of TcpP and ToxT, activates and represses transcription of the genes encoding two outer-membrane porins, OmpU and OmpT. To determine the role of ToxR-dependent porin regulation in V. cholerae pathogenesis, the ToxR-activated ompU promoter was used to drive ompT transcription in a strain lacking OmpU. Likewise, the ToxR-repressed ompT promoter was used to drive ompU transcription in a strain lacking both ToxR and OmpT. This strategy allowed the generation of a toxR+ strain that expresses OmpT in place of OmpU, and a toxR− strain that expresses OmpU in place of OmpT. Growth rates in the presence of bile salts and other anionic detergents were retarded for the toxR+ V. cholerae expressing OmpT in place of OmpU, but increased in toxR− V. cholerae expressing OmpU in place of OmpT. Additionally, the toxR+ V. cholerae expressing OmpT in place of OmpU expressed less cholera toxin and toxin-coregulated pilus, and this effect was shown to be caused by reduced toxT transcription in this strain. Finally, the toxR+ V. cholerae expressing OmpT in place of OmpU was ≈100-fold reduced in its ability to colonize the infant-mouse intestine. Our results indicate that ToxR-dependent modulation of the outer membrane porins OmpU and OmpT is critical for V. cholerae bile resistance, virulence factor expression, and intestinal colonization.

A nonnatural transcriptional coactivator

In eukaryotes, sequence-specific DNA-binding proteins activate gene expression by recruiting the transcriptional apparatus and chromatin remodeling proteins to the promoter through protein-protein contacts. In many instances, the connection between DNA-binding proteins and the transcriptional apparatus is established through the intermediacy of adapter proteins known as coactivators. Here we describe synthetic molecules with low molecular weight that act as transcriptional coactivators. We demonstrate that a completely nonnatural activation domain in one such molecule is capable of stimulating transcription in vitro and in vivo. The present strategy provides a means of gaining external control over gene activation through intervention using small molecules.

Mendel-GFDb and Mendel-ESTS: databases of plant gene families and ESTs annotated with gene family numbers and gene family names

There is no control over the information provided with sequences when they are deposited in the sequence databases. Consequently mistakes can seed the incorrect annotation of other sequences. Grouping genes into families and applying controlled annotation overcomes the problems of incorrect annotation associated with individual sequences. Two databases (http://www.mendel.ac.uk) were created to apply controlled annotation to plant genes and plant ESTs: Mendel-GFDb is a database of plant protein (gene) families based on gapped-BLAST analysis of all sequences in the SWISS-PROT family of databases. Sequences are aligned (ClustalW) and identical and similar residues shaded. The families are visually curated to ensure that one or more criteria, for example overall relatedness and/or domain similarity relate all sequences within a family. Sequence families are assigned a ‘Gene Family Number’ and a unified description is developed which best describes the family and its members. If authority exists the gene family is assigned a ‘Gene Family Name’. This information is placed in Mendel-GFDb. Mendel-ESTS is primarily a database of plant ESTs, which have been compared to Mendel-GFDb, completely sequenced genomes and domain databases. This approach associated ESTs with individual sequences and the controlled annotation of gene families and protein domains; the information being placed in Mendel-ESTS. The controlled annotation applied to genes and ESTs provides a basis from which a plant transcription database can be developed.

Forebrain mechanisms of nociception and pain: Analysis through imaging

Pain is a unified experience composed of interacting discriminative, affective-motivational, and cognitive components, each of which is mediated and modulated through forebrain mechanisms acting at spinal, brainstem, and cerebral levels. The size of the human forebrain in relation to the spinal cord gives anatomical emphasis to forebrain control over nociceptive processing. Human forebrain pathology can cause pain without the activation of nociceptors. Functional imaging of the normal human brain with positron emission tomography (PET) shows synaptically induced increases in regional cerebral blood flow (rCBF) in several regions specifically during pain. We have examined the variables of gender, type of noxious stimulus, and the origin of nociceptive input as potential determinants of the pattern and intensity of rCBF responses. The structures most consistently activated across genders and during contact heat pain, cold pain, cutaneous laser pain or intramuscular pain were the contralateral insula and anterior cingulate cortex, the bilateral thalamus and premotor cortex, and the cerebellar vermis. These regions are commonly activated in PET studies of pain conducted by other investigators, and the intensity of the brain rCBF response correlates parametrically with perceived pain intensity. To complement the human studies, we developed an animal model for investigating stimulus-induced rCBF responses in the rat. In accord with behavioral measures and the results of human PET, there is a progressive and selective activation of somatosensory and limbic system structures in the brain and brainstem following the subcutaneous injection of formalin. The animal model and human PET studies should be mutually reinforcing and thus facilitate progress in understanding forebrain mechanisms of normal and pathological pain.

Event-related brain potentials in the study of visual selective attention

Event-related brain potentials (ERPs) provide high-resolution measures of the time course of neuronal activity patterns associated with perceptual and cognitive processes. New techniques for ERP source analysis and comparisons with data from blood-flow neuroimaging studies enable improved localization of cortical activity during visual selective attention. ERP modulations during spatial attention point toward a mechanism of gain control over information flow in extrastriate visual cortical pathways, starting about 80 ms after stimulus onset. Paying attention to nonspatial features such as color, motion, or shape is manifested by qualitatively different ERP patterns in multiple cortical areas that begin with latencies of 100–150 ms. The processing of nonspatial features seems to be contingent upon the prior selection of location, consistent with early selection theories of attention and with the hypothesis that spatial attention is “special.”

Novel polymers: Molecular to nanoscale order in three dimensions

The assembly of polymer chains in solution is a powerful method that is leading to the preparation of interesting and unique macromolecular-based synthetic nanostructures. Specific control over the intramolecular and intermolecular physical interactions dictates either the folding of single chains or the aggregation and ordering of multiple chains. This control is provided through the selective placement of functional groups along the polymer backbone and the relative strengths of their attractive and repulsive interactions.

Efeitos da probabilidade e proporção de reforço sobre o comportamento de escolha em ratos Wistar (Rattus norvegicus)

A metacognição é processo conceituado como o julgamento que um organismo (humano ou não humano) faz sobre seu próprio saber ou não saber. Há relatos de pesquisas sobre esse processo com seres humanos e diversas espécies de não humanos. Poucos estudos, porém, discutem a ocorrência de metacognição em ratos, e os resultados são controversos, em função de questionamentos sobre os procedimentos experimentais empregados. Este estudo teve o objetivo de investigar o efeito da manipulação de diferentes proporções de reforço produzidas em duas alternativas, sendo uma probabilística e outra com reforçamento contínuo, sobre o desempenho de ratos em uma tarefa de discriminação de diferentes durações de estímulos sonoros. O procedimento empregado é uma adaptação do utilizado por Foote e Crystal (2007), que investigou a ocorrência de metacognição em ratos. Foram utilizados cinco ratos machos, da linhagem Wistar (Rattus norvegicus) mantidos a 80% de seu peso ad libitum. O aparato utilizado foi um labirinto em Ey. O procedimento consistiu de quatro fases: 1) Treino exploratório no braço em Y, no qual o animal foi exposto a alternativas que continham seis pelotas de ração; 2) Treino de discriminação de estímulos sonoros, no qual foram treinadas duas discriminações condicionais com duas durações de estímulo sonoro, uma curta (2s), e uma longa (8s), cada uma correlacionada com a escolha de uma das portas do braço em Y; 3) Treino exploratório no braço em I, no qual os animais foram expostos a uma alternativa livre, que continha três pelotas de ração; e 4) Fase de Teste, na qual foram apresentadas diferentes durações de som (2.00, 2.44, 2.97, 3.62, 4.42, 5.38, 6.56 e 8.00s), a partir das quais o animal poderia escolher entre o braço em Y (fazer o teste), e receber seis pelotas de ração caso escolhesse a porta correta (correlacionada à duração curta ou longa), ou escolher a alternativa de recusa do teste, produzindo, com certeza, a quantidade de ração estabelecida pela condição em vigor. Foi analisada a porcentagem de escolhas realizadas pelos animais nos braços Y e I em cada condição, assim como a relação entre a porcentagem de acertos e erros nos testes e recusa, para cada duração de som. Todos os sujeitos atingiram o critério de aprendizagem estabelecido na fase de treino. Na fase de testes, observou-se que o som deixou de exercer controle sobre a resposta de escolha de todos os animais. À medida que a proporção de reforço variou na alternativa de recusa, os animais alteraram o padrão de escolha, de propensão para aversão ao risco, de acordo com a condição em vigor. A escolha por uma alternativa não se mostrou sob controle da acurácia dos animais em discriminar as durações dos estímulos apresentados, mas sim da proporção e probabilidade do reforço em cada alternativa. Discute-se a necessidade de se recorrer ao conceito de metacognição para descrever o desempenho dos animais em tarefas como a empregada no presente estudo.

Investigação de aquífero fraturado para entendimento de fluxo e transporte de contaminantes clorados: estudo de caso em Valinhos, SP

Casos de contaminação de aquíferos fraturados são bastante complexos, tendo em vista a heterogeneidade das redes de fraturas, e no geral, sua investigação demanda a utilização de técnicas pouco usuais, como por exemplo o imageamento acústico e a perfilagem de velocidade de fluxo de água. Na área de estudo, localizada em Valinhos/SP, o uso inadequado de solventes organoclorados no passado ocasionou a contaminação do aquífero raso em duas áreas, e o aparecimento de concentrações no aquifero profundo levaram a condução do atual trabalho, que teve como principal objetivo a elaboração de um modelo conceitual de fluxo de água e transporte de contaminantes no aquífero cristalino. Previamente à investigação do aquífero fraturado, foi realizada uma análise de trabalhos existentes, incluindo a interpretação de lineamentos, levantamentos geológicos além de perfilagens geofísicas de superfície. Em cada área investigada, foi realizada a perfuração de um poço profundo e aplicadas as técnicas de perfilagens de raios gama, cáliper, flowmeter, imageamento acústico, além da filmagem do poço e realização de ensaios hidráulicos nos dois pontos perfurados. Para caracterização química do aquífero fraturado, foram realizadas coletas de água subterrânea em intervalos selecionados com a utilização de obturadores pneumáticos. As cargas hidráulicas medidas durante a amostragem também auxiliaram no entendimento da direção do fluxo de água. O aquífero cristalino é formado por rochas gnáissicas e se encontra bastante fraturado e intemperizado, principalmente na porção superficial da rocha (até aproximadamente 65,0 m) onde as maiores velocidades de fluxo de água também foram observadas. A rocha sã possui uma menor densidade de fraturas e predominância de minerais mais claros. As fraturas de baixo a médio angulo de mergulho (Grupo 1) são as mais frequentes em ambas as perfurações e possuem direção principal N-S a NE-SW. São observadas, no geral, exercendo grande influência sobre o fluxo de água, principalmente na porção alterada do gnaisse. Fraturas com ângulo elevado de mergulho, classificadas como Grupo 2 (paralelas à foliação) e Grupo 3 (direção NW à W), são também observadas ao longo de toda a perfuração estabelecendo a conexão hidráulica entre as fraturas do Grupo 1. Em menor proporção, são ainda verificadas fraturas com ângulos de mergulho >40 ° pertencente aos Grupos 4 (NE-SW), 5 (E-W), 6 (NW-SE) e 7 (E-W). O fluxo de água subterrânea se mostrou descendente na porção superior da rocha alterada e ascendente na porção mais profunda, possivelmente direcionando a água subterrânea para a região de transição da rocha mais alterada para a rocha sã (entre 61 a 65 m de profundidade). Apesar do fluxo ascendente em profundidade, o bombeamento de poços tubulares existentes no entorno ao longo dos anos, favoreceu a migração dos contaminantes para porções mais profundas. Os contaminantes observados no poço tubular P6 possuem maior semelhança com os contaminantes observados na Área 2, e ambos estão localizados entre lineamentos NW-SE, indicando uma possível influência dos lineamentos no controle sobre o fluxo de água. No entanto, para entendimento do transporte dos contaminantes em área, é necessário um adensamento da rede de monitoramento, levando em consideração a heterogeneidade do meio e as incertezas relacionadas à extrapolação dos dados para áreas não investigadas.

A Fair Day's Wages: Liberty, Legality, and Liability Among Denver's Day Laborers

Day laborers occupy an essential position in Denver’s booming construction industry. Day laborers make up a highly flexible, highly effective workforce able to respond to market changes. For day laborers, informal day-labor gathering points provide increased control over working hours and employee-employer relationships when compared to traditional wage labor. Still, recent legislation and policies around irregular migration has forced large numbers of workers who may have benefited from the stability of full-time regular employment into the informal sector. The day laborers’ flexibility also exposes them to employers constantly inventing ways to deny them the wages and benefits they are owed. Despite changes in Colorado law in attempts to strengthen workers’ recourse against their employers, and despite social and individual tactics day laborers employ to mitigate their vulnerability, systematic structural, symbolic, and everyday violence continue to advantage employers.

When Wells Run Dry: Water and Tourism Along the Western Coast of Nicaragua

This dissertation uses a political ecology approach to examine the relationship between tourism development and groundwater in southwest Nicaragua. Tourism in Nicaragua is a booming industry bolstered by ‘unspoiled’ natural beauty, low crime rates, and government incentives. This growth has led to increased infrastructure, revenue, and employment opportunities for many local communities along the Pacific coast. Not surprisingly, it has also brought concomitant issues of deeper poverty, widening gaps between rich and poor, and competition over natural resources. Adequate provisions of freshwater are necessary to sustain the production and reproduction of tourism; however, it remains uncertain if groundwater supplies can keep pace with demand. The objective of this research is to assess water supply availability amidst tourism development in the Playa Gigante area. It addresses the questions: 1) are local groundwater supplies sufficient to sustain the demand for freshwater imposed by increased tourism development? and 2) is there a power relationship between tourism development and control over local freshwater that would prove inequitable to local populations? Integrating the findings of groundwater monitoring, geological mapping, and ethnographic and survey research from a representative stretch of Pacific coastline, this dissertation shows that diminishing recharge and increased groundwater consumption is creating conflict between stakeholders with various levels of knowledge, power, and access. Although national laws are structured to protect the environment and ensure equitable access to groundwater, the current scramble to secure water has powerful implications on social relations and power structures associated with tourism development. This dissertation concludes that marginalization due to environmental degradation is attributable to the nexus of a political promotion of tourism, poorly enforced state water policies, insufficient water research, and climate change. Greater technical attention to hydrological dynamics and collaboration amongst stakeholders are necessary for equitable access to groundwater, environmental sustainability, and profitability of tourism.

Restaurant Health and Hygiene: A Model Program with Theory-Based Guidelines for Implementation

Poor hygienic practices and illness of restaurant employees are major contributors to the contamination of food and the occurrence of food-borne illness in the United States, costing the food industry and society billions of dollars each year. Risk factors associated with this problem include lack of proper handwashing; food handlers reporting to work sick; poor personal hygiene; and bare hand contact with ready-to-eat foods. However, traditional efforts to control these causes of food-borne illness by public health authorities have had limited impact, and have revealed the need for comprehensive and innovative programs that provide active managerial control over employee health and hygiene in restaurant establishments. Further, the introduction and eventual adoption by the food industry of such programs can be facilitated through the use of behavior-change theory. This Capstone Project develops a model program to assist restaurant owners and operators in exerting active control over health and hygiene in their establishments and provides theory-based recommendations for the introduction of the program to the food industry.

Stochastic choice analysis of tourism destinations

The analysis of tourist destination choice, defined by intra-country administrative units and by product types “coastal/inland and village/city”, permits the characterisation of tourist flow behaviour, which is fundamental for public planning and business management. In this study, we analyse the determinant factors of tourist destination choice, proposing various research hypotheses relative to the impact of destination attributes and the personal characteristics of tourists. The methodology applied estimates Nested and Random Coefficients Multinomial Logit Models, which allow control over possible correlations among different destinations. The empirical application is realised in Spain on a sample of 3,781 individuals and allows us to conclude that prices, distance to the destination and personal motivations are determinants in destination choice.

Open-access operating algorithms for commercial videokeratographer and improvement of corneal sampling

We present an algorithm to process images of reflected Placido rings captured by a commercial videokeratoscope. Raw data are obtained with no Cartesian-to-polar-coordinate conversion, thus avoiding interpolation and associated numerical artifacts. The method provides a characteristic equation for the device and is able to process around 6 times more corneal data than the commercial software. Our proposal allows complete control over the whole process from the capture of corneal images until the computation of curvature radii.

Intelligent Decision-Making System Frameworks for a Digital Plan of Work: A Theoretical Investigation for the Construction Industry

The construction industry is characterised by fragmentation and suffers from lack of collaboration, often adopting adversarial working practices to achieve deliverables. For the UK Government and construction industry, BIM is a game changer aiming to rectify this fragmentation and promote collaboration. However it has become clear that there is an essential need to have better controls and definitions of both data deliverables and data classification. Traditional methods and techniques for collating and inputting data have shown to be time consuming and provide little to improve or add value to the overall task of improving deliverables. Hence arose the need in the industry to develop a Digital Plan of Work (DPoW) toolkit that would aid the decision making process, providing the required control over the project workflows and data deliverables, and enabling better collaboration through transparency of need and delivery. The specification for the existing Digital Plan of Work (DPoW) was to be, an industry standard method of describing geometric, requirements and data deliveries at key stages of the project cycle, with the addition of a structured and standardised information classification system. However surveys and interviews conducted within this research indicate that the current DPoW resembles a digitised version of the pre-existing plans of work and does not push towards the data enriched decision-making abilities that advancements in technology now offer. A Digital Framework is not simply the digitisation of current or historic standard methods and procedures, it is a new intelligent driven digital system that uses new tools, processes, procedures and work flows to eradicate waste and increase efficiency. In addition to reporting on conducted surveys above, this research paper will present a theoretical investigation into usage of Intelligent Decision Support Systems within a digital plan of work framework. Furthermore this paper will present findings on the suitability to utilise advancements in intelligent decision-making system frameworks and Artificial Intelligence for a UK BIM Framework. This should form the foundations of decision-making for projects implemented at BIM level 2. The gap identified in this paper is that the current digital toolkit does not incorporate the intelligent characteristics available in other industries through advancements in technology and collation of vast amounts of data that a digital plan of work framework could have access to and begin to develop, learn and adapt for decision-making through the live interaction of project stakeholders.

Política y criminalidad en el «nuevo Estado» franquista. La criminalización del «enemigo» en el Derecho penal de posguerra

La consolidación del «nuevo Estado» franquista conllevó la elaboración de una doctrina y una legislación penal que permitiera reprimir a los enemigos políticos de la dictadura, así como conseguir el control de la sociedad. Este estudio analiza la criminalización de la desviación social durante la posguerra. La criminalización es la selección de un grupo de personas, a las que el poder del Estado somete mediante la coacción punitiva de sus acciones. En este proceso de «construcción social» del delito, destacó la categorización del «otro» como un enemigo peligroso y dañino moral y socialmente, también en la justicia común en España.