Laribacter hongkongensis: a potential cause of infectious diarrhea

In this study, we describe the isolation of Laribacter hongkongensis, a recently described genus and species of bacterium, in pure culture on charcoal cefoperazone deoxycholate agar from the stool of six patients with diarrhea. Three patients were residents of Hong Kong, and three of Switzerland. In none of the stool samples obtained from these six patients was Salmonella, Shigella, enterohemorrhagic Escherichia coli, Vibrio, Aeromonas, Plesiomonas, or Campylobacter recovered. Rotavirus antigen detection, electron microscopic examination for viruses, and microscopic examinations for ova and cysts were all negative for the stool samples obtained from the three patients in Hong Kong. Enterotoxigenic E. coli was recovered from one of the patients in Hong Kong. Unlike L. hongkongensis type strain HKU1, all the six strains were motile with bipolar flagellae. Sequencing of the 16S ribosomal RNA genes of the six strains showed that they all had sequences with only 0-2 base differences to that of the type strain. Pulsed field gel electrophoresis of the SpeI digested genomic DNA of the six isolates and that of the type strain revealed that the seven isolates were genotypically unrelated strains. More extensive epidemiologic studies should be carried out to ascertain the causative association between L. hongkongensis and diarrhea and to define the reservoir and modes of transmission of L. hongkongensis.

Sex differences in the prevalence and clinical outcomes of subclinical peripheral artery disease in the Health, Aging, and Body Composition (Health ABC) study

The objective of the study was to determine if there are sex-based differences in the prevalence and clinical outcomes of subclinical peripheral artery disease (PAD). We evaluated the sex-specific associations of ankle-brachial index (ABI) with clinical cardiovascular disease outcomes in 2797 participants without prevalent clinical PAD and with a baseline ABI measurement in the Health, Aging, and Body Composition study. The mean age was 74 years, 40% were black, and 52% were women. Median follow-up was 9.37 years. Women had a similar prevalence of ABI < 0.9 (12% women versus 11% men; P = 0.44), but a higher prevalence of ABI 0.9-1.0 (15% versus 10%, respectively; P < 0.001). In a fully adjusted model, ABI < 0.9 was significantly associated with higher coronary heart disease (CHD) mortality, incident clinical PAD and incident myocardial infarction in both women and men. ABI < 0.9 was significantly associated with incident stroke only in women. ABI 0.9-1.0 was significantly associated with CHD death in both women (hazard ratio 4.84, 1.53-15.31) and men (3.49, 1.39-8.72). However, ABI 0.9-1.0 was significantly associated with incident clinical PAD (3.33, 1.44-7.70) and incident stroke (2.45, 1.38-4.35) only in women. Subclinical PAD was strongly associated with adverse CV events in both women and men, but women had a higher prevalence of subclinical PAD.

An improved simple rat model for global cerebral ischaemia by induced cardiac arrest

OBJECTIVES Cerebral hypoxic-ischaemic injury following cardiac arrest is a devastating disease affecting thousands of patients each year. There is a complex interaction between post-resuscitation injury after whole-body ischaemia-reperfusion and cerebral damage which cannot be explored in in vitro systems only; there is a need for animal models. In this study, we describe and evaluate the feasibility and efficiency of our simple rodent cardiac arrest model. METHODS Ten wistar rats were subjected to 9 and 10 minutes of cardiac arrest. Cardiac arrest was introduced with a mixture of the short-acting beta-blocking drug esmolol and potassium chloride. RESULTS All animals could be resuscitated within 1 minute, and survived until day 5.General health score and neurobehavioural testing indicated substantial impairment after cardiac arrest, without differences between groups. Histological examination of the hippocampus CA1 segment, the most vulnerable segment of the cerebrum, demonstrated extensive damage in the cresyl violet staining, as well as in the Fluoro-Jade B staining and in the Iba-1 staining, indicating recruitment of microglia after the hypoxic-ischaemic event. Again, there were no differences between the 9- and 10-minute cardiac arrest groups. DISCUSSION We were able to establish a simple and reproducible 9- and 10-minute rodent cardiac arrest models with a well-defined no-flow-time. Extensive damage can be found in the hippocampus CA1 segment. The lack of difference between 9- and 10-minute cardiac arrest time in the neuropsychological, the open field test and the histological evaluations is mainly due to the small sample size.

Early brain injury linearly correlates with reduction in cerebral perfusion pressure during the hyperacute phase of subarachnoid hemorrhage

BACKGROUND It is unclear how complex pathophysiological mechanisms that result in early brain injury (EBI) after subarachnoid hemorrhage (SAH) are triggered. We investigate how peak intracranial pressure (ICP), amount of subarachnoid blood, and hyperacute depletion of cerebral perfusion pressure (CPP) correlate to the onset of EBI following experimental SAH. METHODS An entire spectrum of various degrees of SAH severities measured as peak ICP was generated and controlled using the blood shunt SAH model in rabbits. Standard cardiovascular monitoring, ICP, CPP, and bilateral regional cerebral blood flow (rCBF) were continuously measured. Cells with DNA damage and neurodegeneration were detected using terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) and Fluoro-jade B (FJB). RESULTS rCBF was significantly correlated to reduction in CPP during the initial 15 min after SAH in a linear regression pattern (r (2) = 0.68, p < 0.001). FJB- and TUNEL-labeled cells were linearly correlated to reduction in CPP during the first 3 min of hemorrhage in the hippocampal regions (FJB: r (2) = 0.50, p < 0.01; TUNEL: r (2) = 0.35, p < 0.05), as well as in the basal cortex (TUNEL: r (2) = 0.58, p < 0.01). EBI occurred in animals with severe (relative CPP depletion >0.4) and moderate (relative CPP depletion >0.25 but <0.4) SAH. Neuronal cell death was equally detected in vulnerable and more resistant brain regions. CONCLUSIONS The degree of EBI in terms of neuronal cell degeneration in both the hippocampal regions and the basal cortex linearly correlates with reduced CPP during hyperacute SAH. Temporary CPP reduction, however, is not solely responsible for EBI but potentially triggers processes that eventually result in early brain damage.

An integrated molecular biology and computational approach to CYP1A1 expression in human brain

The cytochromes P450 comprise a superfamily of heme-containing mono-oxygenases. These enzymes metabolize numerous xenobiotics, but also play a role in metabolism of endogenous compounds. The P450 1A1 enzyme generally metabolizes polycyclic aromatic hydrocarbons, and its expression can be induced by aryl hydrocarbon receptor (AhR) activation. CYP1A1 is an exception to the generality that the majority of CYPs demonstrate highest expression in liver; CYP1Al is present in numerous extrahepatic tissues, including brain. This P450 has been observed in two forms, wildtype (WT) and brain variant (BV), arising from alternatively spliced mRNA transcripts. The CYP1A1 BV mRNA presented an exon deletion and was detected in human brain but not liver tissue of the same individuals. ^ Quantitative PCR analyses were performed to determine CYP1A1 WT and BV transcript expression levels in normal, bipolar disorder or schizophrenic groups. In our samples, we show that CYP1A1 BV mRNA, when present, is found alongside the full-length form. Furthermore, we demonstrate a significant decrease in expression of CYP1A1 in patients with bipolar disorder or schizophrenia. The expression level was not influenced by post-mortem interval, tissue pH, age, tobacco use, or lifetime antipsychotic medication load. ^ There is no indication of increased brain CYP1A1 expression in normal smokers versus non-smokers in these samples. We observed slightly increased CYP1A1 expression only in bipolar and schizophrenic smokers versus non-smokers. This may be indicative of complex interactions between neuronal chemical environments and AhR-mediated CYP1A1 induction in brain. ^ Structural homology modeling demonstrated that P450 1A1 BV has several alterations to positions/orientations of substrate recognition site residues compared to the WT isoform. Automated substrate docking was employed to investigate the potential binding of neurological signaling molecules and neurotropic drugs, as well as to differentiate specificities of the two P450 1A1 isoforms. We consistently observed that the BV isoform produced energetically favorable substrate dockings in orientations not observed for the same substrate in the WT isoform. These results demonstrated that structural differences, namely an expanded substrate access channel and active site, confer greater capacity for unique compound docking positions suggesting a metabolic profile distinct from the wildtype form for these test compounds. ^

Stability and potential application of spliced nuclear pre -mRNA introns

A major portion of this thesis work was dedicated to study the nature and significance of spliced introns. The initial work was focused on studying the IVS1$\sb{\rm C\beta 1}$ intron from a T-cell receptor (TCR)-$\beta$ gene. Compared to an intron lariat control from adenovirus pre-mRNA that was spliced in vitro, IVS1$\sb{\rm C\beta 1}$ was debranched less efficiently by HeLa S100 extracts, although IVS1$\sb{\rm C\beta 1}$ also used the consensus branchpoint in vivo. Subcellular-fractionation analysis showed that most IVS1$\sb{\rm C\beta 1}$ lariats cofractionated with pre-mRNA in the nucleus, consistent with the possibility that intron degradation releases splicing factors which will be available for further rounds of splicing. The half-life of IVS1$\sb{\rm C\beta 1}$ from the endogenous TCR-$\beta$ gene was measured using the general transcription inhibitor actinomycin D to be about $\sim$15 min, which was similar to that of unstable mRNAs such as c-myc mRNA.^ The general transcription inhibitor DRB was also used for intron stability analysis. Unexpectedly, DRB decreased intron and pre-mRNA levels only initially, it later increased the levels of intron-containing RNAs. Inhibition of transcription initiation appeared to be the major early effect (the reduction phase); whereas enhanced premature transcription termination was dominant later (the induction phase).^ Having established the procedures for studying in vivo spliced introns, this approach was applied to study the mechanism of nonsense-mediated downregulation (NMD), a phenomena in which premature termination codons (PTCs) decrease the levels of mRNAs. In this study, the novel intron-oriented approach was applied to study the mechanism of NMD. The levels of spliced introns immediately upstream and downstream of a PTC-bearing exon in a TCR-$\beta$ gene were identified and analyzed along with their pre-mRNA. Although PTC reduced the mRNA levels by 4 to 9 fold, the steady-state levels of spliced introns and the pre-mRNA-to-intron ratios were not significantly altered, indicating that the PTC did not significantly inhibit TCR-$\beta$ RNA splicing. Consistent with this conclusion, the half-lives of the PTC$\sp+$ and PTC$\sp-$ pre-mRNA were similar. The protein synthesis inhibitor cyclohexmide (CHX) upregulated the levels of the PTC$\sp+$ mRNA over 10 fold without affecting the levels of the spliced introns, suggesting that the reversal effect of CHX was through stabilization, not production. These results indicated that inhibition of splicing could not be the major mechanism for the NMD pathway of the TCR-$\beta$ gene, instead, suggesting that mRNA destabilization may be more important. (Abstract shortened by UMI.) ^

Quantitative evaluation of artifact removal in real magnetoencephalogram signals with blind source separation

The magnetoencephalogram (MEG) is contaminated with undesired signals, which are called artifacts. Some of the most important ones are the cardiac and the ocular artifacts (CA and OA, respectively), and the power line noise (PLN). Blind source separation (BSS) has been used to reduce the influence of the artifacts in the data. There is a plethora of BSS-based artifact removal approaches, but few comparative analyses. In this study, MEG background activity from 26 subjects was processed with five widespread BSS (AMUSE, SOBI, JADE, extended Infomax, and FastICA) and one constrained BSS (cBSS) techniques. Then, the ability of several combinations of BSS algorithm, epoch length, and artifact detection metric to automatically reduce the CA, OA, and PLN were quantified with objective criteria. The results pinpointed to cBSS as a very suitable approach to remove the CA. Additionally, a combination of AMUSE or SOBI and artifact detection metrics based on entropy or power criteria decreased the OA. Finally, the PLN was reduced by means of a spectral metric. These findings confirm the utility of BSS to help in the artifact removal for MEG background activity.

Multi-microphone recordings (EigenMike32) inside a WFS system (Iosono) through Ambisonics techniques using Matlab

El audio multicanal ha avanzado a pasos agigantados en los últimos años, y no solo en las técnicas de reproducción, sino que en las de capitación también. Por eso en este proyecto se encuentran ambas cosas: un array microfónico, EigenMike32 de MH Acoustics, y un sistema de reproducción con tecnología Wave Field Synthesis, instalado Iosono en la Jade Höchscule Oldenburg. Para enlazar estos dos puntos de la cadena de audio se proponen dos tipos distintos de codificación: la reproducción de la toma horizontal del EigenMike32; y el 3er orden de Ambisonics (High Order Ambisonics, HOA), una técnica de codificación basada en Armónicos Esféricos mediante la cual se simula el campo acústico en vez de simular las distintas fuentes. Ambas se desarrollaron en el entorno Matlab y apoyadas por la colección de scripts de Isophonics llamada Spatial Audio Matlab Toolbox. Para probar éstas se llevaron a cabo una serie de test en los que se las comparó con las grabaciones realizadas a la vez con un Dummy Head, a la que se supone el método más aproximado a nuestro modo de escucha. Estas pruebas incluían otras grabaciones hechas con un Doble MS de Schoeps que se explican en el proyecto “Sally”. La forma de realizar éstas fue, una batería de 4 audios repetida 4 veces para cada una de las situaciones garbadas (una conversación, una clase, una calle y un comedor universitario). Los resultados fueron inesperados, ya que la codificación del tercer orden de HOA quedo por debajo de la valoración Buena, posiblemente debido a la introducción de material hecho para un array tridimensional dentro de uno de 2 dimensiones. Por el otro lado, la codificación que consistía en extraer los micrófonos del plano horizontal se mantuvo en el nivel de Buena en todas las situaciones. Se concluye que HOA debe seguir siendo probado con mayores conocimientos sobre Armónicos Esféricos; mientras que el otro codificador, mucho más sencillo, puede ser usado para situaciones sin mucha complejidad en cuanto a espacialidad. In the last years the multichannel audio has increased in leaps and bounds and not only in the playback techniques, but also in the recording ones. That is the reason of both things being in this project: a microphone array, EigenMike32 from MH Acoustics; and a playback system with Wave Field Synthesis technology, installed by Iosono in Jade Höchscule Oldenburg. To link these two points of the audio chain, 2 different kinds of codification are proposed: the reproduction of the EigenMike32´s horizontal take, and the Ambisonics´ third order (High Order Ambisonics, HOA), a codification technique based in Spherical Harmonics through which the acoustic field is simulated instead of the different sound sources. Both have been developed inside Matlab´s environment and supported by the Isophonics´ scripts collection called Spatial Audio Matlab Toolbox. To test these, a serial of tests were made in which they were compared with recordings made at the time by a Dummy Head, which is supposed to be the closest method to our hearing way. These tests included other recording and codifications made by a Double MS (DMS) from Schoeps which are explained in the project named “3D audio rendering through Ambisonics techniques: from multi-microphone recordings (DMS Schoeps) to a WFS system, through Matlab”. The way to perform the tests was, a collection made of 4 audios repeated 4 times for each recorded situation (a chat, a class, a street and college canteen or Mensa). The results were unexpected, because the HOA´s third order stood under the Well valuation, possibly caused by introducing material made for a tridimensional array inside one made only by 2 dimensions. On the other hand, the codification that consisted of extracting the horizontal plane microphones kept the Well valuation in all the situations. It is concluded that HOA should keep being tested with larger knowledge about Spherical Harmonics; while the other coder, quite simpler, can be used for situations without a lot of complexity with regards to spatiality.

Behaviour Driven Development for Multi-Agent Systems

This paper presents a testing methodology to apply Behaviour Driven Development (BDD) techniques while developing Multi-Agent Systems (MAS), so called BEhavioural Agent Simple Testing (BEAST) methodology. It is supported by the developed open source framework (BEAST Tool) which automatically generates test cases skeletons from BDD scenarios specifications. The developed framework allows testing MASs based on JADE or JADEX platforms and offers a set of configurable Mock Agents which allow the execution of tests while the system is under development. BEAST tool has been validated in the development of a MAS for fault diagnosis in FTTH (Fiber To The Home) networks.

Estudio preliminar acerca del uso de protocolos y actos comunicativos FIPA en el sistema COMPUTAPLEX

Este trabajo corresponde con la implementación de componentes software dentro de la Plataforma COMPUTAPLEX, la cual tiene como objetivo facilitar a los investigadores la realización de tareas del proceso experimental de ingeniería de software. Uno de los aportes a esta plataforma tecnológica corresponde con el desarrolló de los componentes necesarios para la recuperación de datos experimentales disponibles en diversas fuentes de datos, para ello se hizo uso de un mecanismo capaz de unificar la extracción de información de MySQL, ficheros excel y ficheros SPSS. Con ello diferentes grupos de investigación asociados pueden compartir y tener acceso a repositorios experimentales que se mantienen tanto de manera local como externa. Por otra parte, se ha realizado un estudio de la tecnología de agentes en la que se describe sus definiciones, lenguajes de comunicación, especificación FIPA, JADE como implementación FIPA y parser XML. Además para este trabajo se ha definido e implementado una ontología de comunicación entre agentes, la misma que fue diseñada en la herramienta Protégé. En lo que se refiere al desarrollo de componentes se hizo uso de una amplía variedad de tecnologías que incluye lenguaje de programación Java, framework JADE para el desarrollo de agentes, librería JENA para manejo de ontologías, librería SAXParser para lectura de archivos XML y patrón de diseño Factory. Finalmente se describe la metodología de trabajo utilizada en el proyecto, la cual por medio de la realización de varios ciclos iterativos permitió obtener prototipos que poco a poco fueron cubriendo las necesidades del producto software.----ABSTRACT---- This work relates to the implementation of software components within the platform Computaplex, which aims to enable researchers to conduct experimental software engineering process tasks. One of the contributions to this platform technology corresponds to the development of components which are necessary for the recovery of experimental data available in different data sources, to archive this goal a mechanism able to unify the extraction of information from MySQL, Excel and SPSS files was made. Therefore, associated research groups can share and access experimental repositories that remain both locally and externally. Moreover, it has been conducted a study of agent technology in its definition is described, languages communication, FIPA, JADE and FIPA implementation and XML parser. In addition to this work, it has been defined and implemented an ontology for communication between agents, the same as was designed in the Protégé tool. In what refers to the development of components, a wide range of technologies have been made which includes Java programming language, framework JADE for agent development, JENA library for handling ontologies, SAXParser for reading XML files and Factory design pattern. Finally, describing the work methodology used in this project, which through the implementation of several iterative cycles allowed to obtain prototypes were gradually meeting the needs of the software product.

Tidal and spatial variations of DI13C and aquatic chemistry in a temperate tidal basin during winter time

Here, the pelagic carbonate system and the ?13C signature of dissolved inorganic carbonate (DIC) were investigated in a tidal basin of the southern North Sea, the Jade Bay, with respect to tidal cycles and a transect towards the North Sea in winter time (January and November, 2010). Physical parameters, major and trace elements, and nutrient concentrations were considered, too. Primary production and pelagic organic matter respiration were negligible during winter time. Both, the compositional variations on the transects as well as during the tidal cycles indicate the mixing of North Sea with fresh water. The combined spatial co-variations of different parameters indicate an introduction of fresh water that was enriched in DI12C, metabolites (e.g., ammonia), protons, and dissolved redox-sensitive elements (e.g., Mn2+). During the January campaign, the discharge via the flood gates was limited due to ice cover of the hinterland drainage ditches, allowing for an observation of tidal variations without significant mixing contributions from surface water discharges. Considering a binary mixing model with North Sea and fresh water as end-members, the extrapolated fresh water end-member composition for this campaign is estimated to contain about 3.8 mmol/kg DIC , and enhanced concentrations of NH4+, Mn2+, and protons compared to North Sea water. The fast temporal response of dissolved geochemical tracers on tidal variations in the Jade Bay indicates a continuous supply of a fresh water component. The measured composition of fresh waters entering the Jade Bay via flood gates (end of October, 2010) did not match the values estimated by the binary mixing model. Therefore, the overall fresh water component likely is a mixture between sources originating from flood gates and (in January) dominating submarine groundwater discharge entering the Jade Bay. This model is consistent with the results obtained during the November campaign, when a more important contribution from flood gates is expected and a more variable fresh water end-member is estimated. The co-variations of the concentrations and the stable carbon isotope composition of DIC are applied to evaluate possible superimposed sink-source-transformation processes in the coastal waters and a general co-variation scheme is suggested.