Gnotobiotic IL-10−/−; NF-κBEGFP Mice Develop Rapid and Severe Colitis Following Campylobacter jejuni Infection

Limited information is available on the molecular mechanisms associated with Campylobacter jejuni (C. jejuni) induced food-borne diarrheal illnesses. In this study, we investigated the function of TLR/NF-κB signaling in C. jejuni induced pathogenesis using gnotobiotic IL-10−/−; NF-κBEGFP mice. In vitro analysis showed that C. jejuni induced IκB phosphorylation, followed by enhanced NF-κB transcriptional activity and increased IL-6, MIP-2α and NOD2 mRNA accumulation in infected-mouse colonic epithelial cells CMT93. Importantly, these events were blocked by molecular delivery of an IκB inhibitor (Ad5IκBAA). NF-κB signalling was also important for C.jejuni-induced cytokine gene expression in bone marrow-derived dendritic cells. Importantly, C. jejuni associated IL-10−/−; NF-κBEGFP mice developed mild (day 5) and severe (day 14) ulcerating colonic inflammation and bloody diarrhea as assessed by colonoscopy and histological analysis. Macroscopic analysis showed elevated EGFP expression indicating NF-κB activation throughout the colon of C. jejuni associated IL-10−/−; NF-κBEGFP mice, while fluorescence microscopy revealed EGFP positive cells to be exclusively located in lamina propria mononuclear cells. Pharmacological NF-κB inhibition using Bay 11-7085 did not ameliorate C. jejuni induced colonic inflammation. Our findings indicate that C. jejuni induces rapid and severe intestinal inflammation in a susceptible host that correlates with enhanced NF-κB activity from lamina propria immune cells

Robust estimation of bacterial cell count from optical density

Optical density (OD) is widely used to estimate the density of cells in liquid culture, but cannot be compared between instruments without a standardized calibration protocol and is challenging to relate to actual cell count. We address this with an interlaboratory study comparing three simple, low-cost, and highly accessible OD calibration protocols across 244 laboratories, applied to eight strains of constitutive GFP-expressing E. coli. Based on our results, we recommend calibrating OD to estimated cell count using serial dilution of silica microspheres, which produces highly precise calibration (95.5% of residuals <1.2-fold), is easily assessed for quality control, also assesses instrument effective linear range, and can be combined with fluorescence calibration to obtain units of Molecules of Equivalent Fluorescein (MEFL) per cell, allowing direct comparison and data fusion with flow cytometry measurements: in our study, fluorescence per cell measurements showed only a 1.07-fold mean difference between plate reader and flow cytometry data

Biological Earth observation with animal sensors

Space-based tracking technology using low-cost miniature tags is now delivering data on fine-scale animal movement at near-global scale. Linked with remotely sensed environmental data, this offers a biological lens on habitat integrity and connectivity for conservation and human health; a global network of animal sentinels of environmen-tal change

Cross-linked Phospholipid Coatings in Micron-Sized Channels for Spatially Discrete Bioassays and Protein Separations

Supported phospholipid bilayers (SPBs) are attractive surface coatings since they are resistant to non-specific protein adsorption, self-assemble, support the function of many integral and membrane bound proteins, and can be functionalized with a variety of available headgroup chemistries. Phospholipid capillary coatings have been reported for applications in capillary electrophoresis (CE), although these coatings suffer from inherent stability problems, which arise from the fragility of the interactions between the bilayer and supportive substrate. Patterned immobilized domains can be functionally diversified, enabling for the screening of multiple analytes in a condensed region. The application of polymerized phospholipids was investigated for applications in CE and preparation of low volume biosensing arrays. A polymerized phospholipid, bis-SorbPC, was used to stabilize the phospholipid bilayer through cross-linking on the surface of fused silica capillaries and compared to other phospholipid capillary coatings for stability and separation quality. Polymerized bis-SorbPC coatings have significantly longer stability against degradation and can be stored for > one year with improved separation efficiency of proteins over bare capillary separations.Three-dimensional fluidic channels were modified with polymerized phospholipid features, which can be chemical "tuned" to introduce specific affinity binding, such as biotin-avidin or antibody-antigen binding, while minimizing nonspecific protein binding to the area of interest. Patterning of bis-SorbPC channels was achieved and the void regions between poly(bis-SorbPC) regions can be filled with other phospholipids to monitor the non-covalent binding (e.g. biotin-avidin, NTA-Ni2+-6xHis-protein) and covalent binding of proteins to DOPE-PEG-pNP and maleimide functionalized lipids, while minimizing non-specific adsorption and protein denaturation. Analytes can be immobilized on the surface via immunobinding events for detection and immunoassay applications. SPBs were used to support membrane proteins within patterned regions of the channel. Functionality can be added to the poly(bis-SorbPC) regions with copolymerization allowing regions of distinct functionality to reside adjacently in the channel. Through multiple polymerization steps, channels with multiple functionalities are demonstrated, with the possibility for higher-throughput multi-functional biosensing channels in the future

Virkning på sporslitasje ved etablering av retningsseparerende tiltak på tofelts riksveger i Statens vegvesen region midt

Den positive trafikksikkerhetseffekten, av tiltakene forsterket midtoppmerking og midtrekkverk, er blitt godt dokumentert og vil framover være aktuelle tiltak i arbeidet mot møteulykkene. Derimot har effekten av tiltakene i form av redusert dekkelevetid ikke fått et like stort fokus. Formålet med denne masteroppgaven har vært å undersøke dekkelevetiden på bakgrunn av konsekvensene, som midtmarkeringen medfører i form av økt kanalisert trafikk og med det økt sporutvikling. I tilknytning til undersøkelsen ble det gjennom en litteraturstudie gjennomgått teorier og tidligere undersøkelser på området. Problemstillingen i oppgaven ble uttrykt med hypotesen om at sporparameternes årlige sporutvikling er like før og etter tiltak med forsterket midtoppmerking eller midtrekkverk. For å besvare problemstillingen ble 18 strekninger fra Region midt undersøkt. Strekningene som ble undersøkt var 2-felts veger, som ikke hadde fått breddeutvidelse i forbindelsen med tiltaket. Oppgavens empiriske del var hovedsakelig spordata, i form av detaljdata, fra Nasjonal VegDataBase (NVDB) og dekkeopplysninger fra dekkeplanleggingsverktøyet PMS 2010. For å finne ut om sporutviklingen ble endret etter tiltak, ble strekningene undersøkt statistisk basert på spordata før og etter tiltak. Hovedtrendene fra undersøkelsen viste at tiltakene påvirket trafikkens plassering i kjørefeltet, ved at sporbredden ble smalere og gav økt sporutvikling. ÅDT og dekkekvaliteten viste seg å ha stor betydning for sporutviklingen. Siden parameterne til strekningene hadde ulike verdier, gav dette en usikkerhet i resultatene, som gjorde at en ikke oppnådde formålet en hadde til hensikt å undersøke. Det er mulig at resultatene ville ha vært mer pålitelige, om man hadde analysert sporendringen direkte på 20 m-verdiene som ligger til grunn. Et større nedslagsfelt med tanke på valg av strekninger, hadde sannsynligvis gitt flere sammenlignbare strekninger og en mer pålitelig konklusjon

Bubble-Point Measurements of <i>n</i>‑Butane + <i>n</i>‑Octane and <i>n</i>‑Butane + <i>n-</i>Nonane Binary Mixtures

Mixtures of small gaseous hydrocarbons with longer chain hydrocarbons are of interest to the natural gas industry as well as other industries in which separations are critical. In particular, binary mixtures of <i>n-</i>nonane are of interest, because <i>n-</i>nonane was recently incorporated into the GERG-2008 equation of state, but there is little experimental vapor–liquid equilibrium (VLE) data available to support the equation. The bubble-point pressures of four compositions of each of the binary mixtures <i>n</i>-butane + <i>n</i>-octane and <i>n</i>-butane + <i>n-</i>nonane were measured over the temperature range of 270 to 370 K. The data and the expanded uncertainty (at a 95 % confidence level, <i>k</i> = 2) of each point are reported. Additionally, the data are compared to existing literature data for the <i>n-</i>butane + <i>n</i>-octane and the GERG-2008 equation for both systems. This is the first report of vapor–liquid equilibrium measurements on <i>n</i>-butane + <i>n-</i>nonane binary mixtures

Bubble-Point Measurements and Modeling of Binary Mixtures of Linear Siloxanes

The bubble-point pressures of three binary mixtures of linear siloxanes have been measured. The binary mixtures consist of hexamethyldisiloxane (MM) which is mixed with either octamethyltrisiloxane (MDM), decamethyltetrasiloxane (MD2M), and dodecamethylpentasiloxane (MD3M). For each mixture, three compositions were measured in which MM was present in approximately 25 mol %, 50 mol %, and 75 mol %. The bubble-point pressures were measured over a temperature range of 270 to 380 K for all mixtures. Large uncertainties are observed for the lower temperatures (below 320 K) due to noncondensable impurities. A detailed analysis is performed to determine the effect of noncondensable gases on the measured bubble-point pressure data. The newly obtained bubble-point pressure data is used to determine new binary interaction parameters for the multicomponent Helmholtz energy model. The data used for the fitting of the binary interaction parameters are weighted by the relative uncertainty; this ensures that data points with high uncertainty contribute less to the final binary interaction parameter. In this work, a description of the experimental apparatus and measurement procedure is given, as well as the measured bubble-point pressure data and newly obtained binary interaction parameters

GCD capacitance

Capacitance obtained from galvanostatic charge discharge curves at varying current rates