Identification of infectious agents in onychomycoses by PCR-terminal restriction fragment length polymorphism.

A fast and reliable assay for the identification of dermatophyte fungi and nondermatophyte fungi (NDF) in onychomycosis is essential, since NDF are especially difficult to cure using standard treatment. Diagnosis is usually based on both direct microscopic examination of nail scrapings and macroscopic and microscopic identification of the infectious fungus in culture assays. In the last decade, PCR assays have been developed for the direct detection of fungi in nail samples. In this study, we describe a PCR-terminal restriction fragment length polymorphism (TRFLP) assay to directly and routinely identify the infecting fungi in nails. Fungal DNA was easily extracted using a commercial kit after dissolving nail fragments in an Na(2)S solution. Trichophyton spp., as well as 12 NDF, could be unambiguously identified by the specific restriction fragment size of 5'-end-labeled amplified 28S DNA. This assay enables the distinction of different fungal infectious agents and their identification in mixed infections. Infectious agents could be identified in 74% (162/219) of cases in which the culture results were negative. The PCR-TRFLP assay described here is simple and reliable. Furthermore, it has the possibility to be automated and thus routinely applied to the rapid diagnosis of a large number of clinical specimens in dermatology laboratories.

Synthesis of a radioiodinated photoreactive MAGE-1 peptide derivative and photoaffinity labeling of cell-associated human leukocyte antigen-A1 molecules.

The synthesis of a photoreactive derivative of the human leukocyte antigen-A1 (HLA-A1)-restricted MAGE-1 peptide 161-169 (EADPTGHSY) is described. Using conventional automated solid-phase peptide synthesis, a photoreactive derivative of this peptide was synthesized by replacing histidine-167 with photo-reactive N-beta-4-azidosalicyloyl-L-2,3-diaminopropionic acid. The C-terminal tyrosine was incorporated as phosphotyrosine. This peptide derivative was radioiodinated in the presence of chloramine T. This iodination took place selectively at the photoreactive group, because the phosphate ester prevented tyrosine iodination. Following dephosphorylation with alkaline phosphatase and chromatographic purification, the radiolabeled peptide derivative was incubated with cells expressing HLA-A1 or other HLA molecules. Photoactivation resulted in efficient photoaffinity labeling of HLA-A1. Other HLA molecules or other cellular components were not detectably labeled. This labeling was inhibited by HLA-A1 but not by HLA-A2-binding peptides. This synthesis is generally applicable and can also be adapted to the synthesis of well-defined radiolabeled nonphotoreactive peptide derivatives.

Monitoramento Ambiental da Estrada São Domingos – Assomada, Ilha de Santiago, Cabo Verde, com Ênfase na Compartimentação Geomorfológica

Este trabalho tem por objetivo a aferição dos impactos ambientais ocorridos ao longo da estrada São Domingos-Assomada, na ilha de Santiago na República de Cabo Verde, África Ocidental no período de 2003 a 2009, com ênfase sobre os compartimentos geomorfológicos e seus materiais estruturadores, bem como estabelecer medidas de minimização dos referidos impactos. O estudo ainda procurou mostrar que o monitoramento ambiental das obras da rodovia tem uma série de vantagens porque pode identificar tendências nocivas sobre as variáveis ambientais e sociais antes que seja tarde demais para minimizar ou prevenir seus impactos. Ao longo da pesquisa foram destacadas as principais atividades que originaram impactos ambientais causados pelas obras tanto na fase de reabilitação como na fase de exploração da via. Buscou-se ainda mostrar que a crescente intervenção antrópica ao longo da estrada tem repercutido cada vez mais intensamente sobre a dinâmica do relevo e nos processos de erosão do solo associados às práticas inadequadas de uso da terra. Os impactos ambientais ocorridos durante a exploração da via resultaram, sobretudo, na degradação dos compartimentos geomorfológicos, por estes se constituírem no principal suporte das demais organizações espaciais. Metodologicamente este trabalho abrangeu uma pesquisa aplicada de cunho descritivo e exploratório visando melhor identificar as alterações nos sistemas ambientais, decorrentes da implantação da estrada e das atividades antrópicas realizadas no seu entorno. A fim de estimar a repercussão das intervenções junto à população imediatamente afetada foram aplicadas entrevistas voltadas à avaliação da percepção dos riscos ambientais. A coleta de dados geomorfológicos em campo foi realizada mediante a observação in loco das condições ambientais e sua posterior cartografação em gabinete. Os resultados dessas foram cotejados com o levantamento da literatura especializada a fim de permitir o enquadramento coerente do grau dos agravos ambientais analisados. Os resultados do estudo apontaram que a degradação dos compartimentos geomorfológicos e suas formações superficiais não ocorrem unicamente em função da estrada, mas também pelas práticas tradicionais de uso da terra. Ainda foi possível constatar que no caso da estrada São Domingos – Assomada, as obras de recuperação não levam em conta à dinâmica do relevo, ou seja, não consideram os parâmetros morfogenéticos. Dessa forma, tais obras essenciais de infra-estrutura acabam, muitas vezes, durando pouco tempo, desperdiçando recursos financeiros que são escassos nos países em desenvolvimento.

Analysis of acute brain slices by electron microscopy: A correlative light-electron microscopy workflow based on Tokuyasu cryo-sectioning.

Acute brain slices are slices of brain tissue that are kept vital in vitro for further recordings and analyses. This tool is of major importance in neurobiology and allows the study of brain cells such as microglia, astrocytes, neurons and their inter/intracellular communications via ion channels or transporters. In combination with light/fluorescence microscopies, acute brain slices enable the ex vivo analysis of specific cells or groups of cells inside the slice, e.g. astrocytes. To bridge ex vivo knowledge of a cell with its ultrastructure, we developed a correlative microscopy approach for acute brain slices. The workflow begins with sampling of the tissue and precise trimming of a region of interest, which contains GFP-tagged astrocytes that can be visualised by fluorescence microscopy of ultrathin sections. The astrocytes and their surroundings are then analysed by high resolution scanning transmission electron microscopy (STEM). An important aspect of this workflow is the modification of a commercial cryo-ultramicrotome to observe the fluorescent GFP signal during the trimming process. It ensured that sections contained at least one GFP astrocyte. After cryo-sectioning, a map of the GFP-expressing astrocytes is established and transferred to correlation software installed on a focused ion beam scanning electron microscope equipped with a STEM detector. Next, the areas displaying fluorescence are selected for high resolution STEM imaging. An overview area (e.g. a whole mesh of the grid) is imaged with an automated tiling and stitching process. In the final stitched image, the local organisation of the brain tissue can be surveyed or areas of interest can be magnified to observe fine details, e.g. vesicles or gold labels on specific proteins. The robustness of this workflow is contingent on the quality of sample preparation, based on Tokuyasu's protocol. This method results in a reasonable compromise between preservation of morphology and maintenance of antigenicity. Finally, an important feature of this approach is that the fluorescence of the GFP signal is preserved throughout the entire preparation process until the last step before electron microscopy.

Atlas-based segmentation of pathological MR brain images using a model of lesion growth.

We propose a method for brain atlas deformation in the presence of large space-occupying tumors, based on an a priori model of lesion growth that assumes radial expansion of the lesion from its starting point. Our approach involves three steps. First, an affine registration brings the atlas and the patient into global correspondence. Then, the seeding of a synthetic tumor into the brain atlas provides a template for the lesion. The last step is the deformation of the seeded atlas, combining a method derived from optical flow principles and a model of lesion growth. Results show that a good registration is performed and that the method can be applied to automatic segmentation of structures and substructures in brains with gross deformation, with important medical applications in neurosurgery, radiosurgery, and radiotherapy.

Brain structure in movement disorders: a neuroimaging perspective.

Purpose of review: An overview of recent advances in structural neuroimaging and their impact on movement disorders research is presented. Recent findings: Novel developments in computational neuroanatomy and improvements in magnetic resonance image quality have brought further insight into the pathophysiology of movement disorders. Sophisticated automated techniques allow for sensitive and reliable in-vivo differentiation of phenotype/genotype related traits and their interaction even at presymptomatic stages of disease. Summary: Voxel-based morphometry consistently demonstrates well defined patterns of brain structure changes in movement disorders. Advanced stages of idiopathic Parkinson's disease are characterized by grey matter volume decreases in basal ganglia. Depending on the presence of cognitive impairment, volume changes are reported in widespread cortical and limbic areas. Atypical Parkinsonian syndromes still pose a challenge for accurate morphometry-based classification, especially in early stages of disease progression. Essential tremor has been mainly associated with thalamic and cerebellar changes. Studies on preclinical Huntington's disease show progressive loss of tissue in the caudate and cortical thinning related to distinct motor and cognitive phenotypes. Basal ganglia volume in primary dystonia reveals an interaction between genotype and phenotype such that brain structure changes are modulated by the presence of symptoms under the influence of genetic factors. Tics in Tourette's syndrome correlate with brain structure changes in limbic, motor and associative fronto-striato-parietal circuits. Computational neuroanatomy provides useful tools for in-vivo assessment of brain structure in movement disorders, allowing for accurate classification in early clinical stages as well as for monitoring therapy effects and/or disease progression.

AWSum-Combining Classification with Knowledge Aquisition

Many classifiers achieve high levels of accuracy but have limited applicability in real world situations because they do not lead to a greater understanding or insight into the^way features influence the classification. In areas such as health informatics a classifier that clearly identifies the influences on classification can be used to direct research and formulate interventions. This research investigates the practical applications of Automated Weighted Sum, (AWSum), a classifier that provides accuracy comparable to other techniques whilst providing insight into the data. This is achieved by calculating a weight for each feature value that represents its influence on the class value. The merits of this approach in classification and insight are evaluated on a Cystic Fibrosis and Diabetes datasets with positive results.

Dynamic social representations of the 2009 H1N1 pandemic: Shifting patterns of sense-making and blame

We investigate dynamics of public perceptions of the 2009 H1N1 influenza pandemic to understand changing patterns of sense-making and blame regarding the outbreak of emerging infectious diseases. We draw on social representation theory combined with a dramaturgical perspective to identify changes in how various collectives are depicted over the course of the pandemic, according to three roles: heroes, villains and victims. Quantitative results based on content analysis of three cross-sectional waves of interviews show a shift from mentions of distant collectives (e.g., far-flung countries) at Wave 1 to local collectives (e.g., risk groups) as the pandemic became of more immediate concern (Wave 2) and declined (Wave 3). Semi-automated content analysis of media coverage shows similar results. Thematic analyses of the discourse associated with collectives revealed that many were consistently perceived as heroes, villains and victims.

Model-based Segmentation and Fusion of 3D Computed Tomography and 3D Ultrasound of the Eye for Radiotherapy Planning

Computed Tomography (CT) represents the standard imaging modality for tumor volume delineation for radiotherapy treatment planning of retinoblastoma despite some inherent limitations. CT scan is very useful in providing information on physical density for dose calculation and morphological volumetric information but presents a low sensitivity in assessing the tumor viability. On the other hand, 3D ultrasound (US) allows a highly accurate definition of the tumor volume thanks to its high spatial resolution but it is not currently integrated in the treatment planning but used only for diagnosis and follow-up. Our ultimate goal is an automatic segmentation of gross tumor volume (GTV) in the 3D US, the segmentation of the organs at risk (OAR) in the CT and the registration of both modalities. In this paper, we present some preliminary results in this direction. We present 3D active contour-based segmentation of the eye ball and the lens in CT images; the presented approach incorporates the prior knowledge of the anatomy by using a 3D geometrical eye model. The automated segmentation results are validated by comparing with manual segmentations. Then, we present two approaches for the fusion of 3D CT and US images: (i) landmark-based transformation, and (ii) object-based transformation that makes use of eye ball contour information on CT and US images.

Automation in clinical bacteriology: what system to choose?

With increased activity and reduced financial and human resources, there is a need for automation in clinical bacteriology. Initial processing of clinical samples includes repetitive and fastidious steps. These tasks are suitable for automation, and several instruments are now available on the market, including the WASP (Copan), Previ-Isola (BioMerieux), Innova (Becton-Dickinson) and Inoqula (KIESTRA) systems. These new instruments allow efficient and accurate inoculation of samples, including four main steps: (i) selecting the appropriate Petri dish; (ii) inoculating the sample; (iii) spreading the inoculum on agar plates to obtain, upon incubation, well-separated bacterial colonies; and (iv) accurate labelling and sorting of each inoculated media. The challenge for clinical bacteriologists is to determine what is the ideal automated system for their own laboratory. Indeed, different solutions will be preferred, according to the number and variety of samples, and to the types of sample that will be processed with the automated system. The final choice is troublesome, because audits proposed by industrials risk being biased towards the solution proposed by their company, and because these automated systems may not be easily tested on site prior to the final decision, owing to the complexity of computer connections between the laboratory information system and the instrument. This article thus summarizes the main parameters that need to be taken into account for choosing the optimal system, and provides some clues to help clinical bacteriologists to make their choice.

Sistema de almacén y dispensación robotizada de productos farmacéuticos

Actualmente, las necesidades de mejora en gestión de stocks y la mayor disponibilidad de sistemas automáticos, están haciendo que muchas empresas inviertan en técnicas modernas para almacenamiento y manipulación de productos. Esta inquietud también ha llegado a las farmacias, que de forma lenta pero firme se van apuntando a su robotización. Uno de los principales problemas a los que se enfrentan las farmacias es la pérdida de tiempo en la gestión y búsqueda de medicamentos, provocando situaciones negativas como las esperas, la falta de tiempo para una atención más personalizada y como consecuencia, la pérdida de clientes. Este inconveniente y la necesidad de mejora en la gestión de los stocks han hecho que aparezcan los Sistemas de dispensación automática de productos farmacéuticos. El dispensador automático facilita el trabajo del farmacéutico al automatizar la búsqueda de la medicina requerida, aumentando la dedicación al cliente y reduciendo los tiempos no productivos y las colas. El presente estudio desarrolla un sistema de dispensación automático de fármacos aplicado a farmacias con una rotación de medicamentos media/ baja, valorando tanto su viabilidad técnica como económica. El almacén propuesto es de tipo caótico con sistema de carga, almacenamiento y descarga completamente automáticos. La mayoría de diseños y conceptos expuestos en este trabajo son de desarrollo propio del autor con el único objetivo de la búsqueda de nuevas soluciones para conseguir un sistema de almacenamiento efectivo y de máximo rendimiento.

Automatització d'un procés de fabricació de peces plàstiques

RESUM L'automatització s'utilitza des de fa molts anys, tot i que va començar a agafar la definició que coneixem actualment al voltant dels anys seixanta i setanta, moment en què es comencen a comercialitzar els primers PLC. A partir d'aquí, el seu creixement ha estat exponencial. En aquest sentit, la tecnologia ha anat avançant i ha augmentat els components que la forma, per això a hores d'ara encara no sabem fins on podrà arribar i què aconseguirà. Per a la indústria tot això ha significat l'automatització de processos que fins ara utilitzaven molt mà d'obra, reduint-la dràsticament. Una de les indústries que més s'ha beneficiat de tots aquests avenços ha estat la de l'automoció, concretament les seves grans línies de producció, automatitzades a uns nivells que fins fa poc temps eren impensables. Aquest projecte forma part d'aquesta indústria, no directament per a la construcció de l'automòbil, sinó indirectament, ja que l'empresa per a la qual s'ha fet l'automatització fabrica peces plàstiques per a automòbils. Concretament, unes peces amb uns injerts metàl•lics conductors que es munten a tots els vehicles i s'utilitzen per accionar els neteja vidres dels cotxes. Aquest fet implica que la fabricació i el disseny de la peça sigui curosament vigilat i controlat per al client final, amb uns controls de qualitat extremadament exigents. El funcionament del procés de fabricació es fa a partir d'unes peces de plàstic produïdes per una injectora que es fan passar per unes estacions automatitzades, cada una de les quals fa una acció concreta per aconseguir el muntatge final.

Realització d'una aplicació de gestió de xarxes per a la xarxa sensefils de Manresa GuifiBages

En els últims anys, la popularitat de les xarxes sensefils (WIFI) ha anat en augment a un ritme incansable. Des de petits aparells instal•lats a les cases amb aquesta tecnologia com a complement dels routers d’accés a internet instal•lats per diverses companyies, fins a empreses fent petits desplegaments per comunicar entre si les seves seus. Al marge d’aquests escenaris, s’ha produït un fenomen social d’acolliment d’aquesta tecnologia a nivell mundial, en forma del que coneixem com a xarxes ciutadanes / xarxes lliures / xarxes socials. Aquestes xarxes han estat possibles gràcies a diverses raons que han fet assequible a col•lectius de persones, tant els aparells com els coneixements necessaris per dur a terme aquestes actuacions. Dintre d’aquest marc, al Bages, concretament a Manresa, es va començar a desenvolupar una d’aquestes xarxes. Les decisions d’aquesta xarxa d’utilitzar exclusivament hardware i software de codi obert, i determinats aspectes tècnics de la xarxa, ha comportat que la xarxa fos incompatible amb algunes de les aplicacions de gestió de xarxes existents desenvolupades per comunicats com gufi.net a Osona. És per això que per garantir el creixement, la supervivència i l’èxit d’aquesta xarxa en el temps, és indispensable poder comptar amb una eina de gestió que s’adigui a les característiques de GuifiBages. L’objectiu principal d’aquest treball és dotar a la xarxa GuifiBages de les eines necessàries per poder gestionar tota la informació referent a l’estructura de la seva xarxa, tant per facilitar l’accés a nous usuaris sense molts coneixements tècnics, com per facilitar nous desplegaments / reparacions / modificacions de la xarxa d’una manera automàtica. Com a conclusió d’aquest treball, podem afirmar que les avantatges que proporciones tecnologies com Plone, faciliten enormement la creació d’aplicacions de gestió de continguts en entorn web. Alhora, l’ús de noves tècniques de programació com AJAX o recursos com els que ofereix Google, permeten desenvolupar aplicacions web que no tenen res a envejar al software tradicional. D’altra banda, voldríem destacar l’ús exclusiu de programari lliure tant en els paquets de software necessaris pel desenvolupament, com en el sistema operatiu i programes dels ordinadors on s’ha dut a terme, demostrant que es poden desenvolupar sistemes de qualitat sense dependre de programari privatiu.

A surface-based approach to quantify local cortical gyrification.

The high complexity of cortical convolutions in humans is very challenging both for engineers to measure and compare it, and for biologists and physicians to understand it. In this paper, we propose a surface-based method for the quantification of cortical gyrification. Our method uses accurate 3-D cortical reconstruction and computes local measurements of gyrification at thousands of points over the whole cortical surface. The potential of our method to identify and localize precisely gyral abnormalities is illustrated by a clinical study on a group of children affected by 22q11 Deletion Syndrome, compared to control individuals.

Gestión centralizada de software de una red sanitaria en Clientes Windows

El proyecto que se presenta trata del estudio y desarrollo para la implementación de un sistema de gestión centralizada de software de una red Sanitaria en Clientes Windows. Este proyecto surge de la necesidad, en una empresa concertada dado el crecimiento de la red, de poder administrar los host de una manera más automatizada y más eficiente.