Host Responses to Intestinal Microbial Antigens in Gluten-Sensitive Mice

Background and Aims

Excessive uptake of commensal bacterial antigens through a permeable intestinal barrier may influence host responses to specific antigen in a genetically predisposed host. The aim of this study was to investigate whether intestinal barrier dysfunction induced by indomethacin treatment affects the host response to intestinal microbiota in gluten-sensitized HLA-DQ8/HCD4 mice.

Methodology/Principal Findings

HLA-DQ8/HCD4 mice were sensitized with gluten, and gavaged with indomethacin plus gluten. Intestinal permeability was assessed by Ussing chamber; epithelial cell (EC) ultra-structure by electron microscopy; RNA expression of genes coding for junctional proteins by Q-real-time PCR; immune response by in-vitro antigen-specific T-cell proliferation and cytokine analysis by cytometric bead array; intestinal microbiota by fluorescence in situ hybridization and analysis of systemic antibodies against intestinal microbiota by surface staining of live bacteria with serum followed by FACS analysis. Indomethacin led to a more pronounced increase in intestinal permeability in gluten-sensitized mice. These changes were accompanied by severe EC damage, decreased E-cadherin RNA level, elevated IFN-γ in splenocyte culture supernatant, and production of significant IgM antibody against intestinal microbiota.

Conclusion

Indomethacin potentiates barrier dysfunction and EC injury induced by gluten, affects systemic IFN-γ production and the host response to intestinal microbiota antigens in HLA-DQ8/HCD4 mice. The results suggest that environmental factors that alter the intestinal barrier may predispose individuals to an increased susceptibility to gluten through a bystander immune activation to intestinal microbiota.     

High-performance liquid chromatographic method for the determination of mangiferin, likviritin and dihydroquercetin in rat plasma and urine

The use of reversed-phase high-performance liquid chromatography for the determination of the biologically active plant phenolic compounds mangiferin, likviritin and dihydroquercetin is described. Perchloric acid (35%) was used for deproteinization in the case of mangiferin and likviritin, and acidified methanol for dihydroquercetin. Detection was performed at 254, 275 and 290 nm for mangiferin, likviritin and dihydroquercetin in plasma, and 365, 312 and 290 nm in urine, respectively. The limit of detection was 0.2 μg/ml for plasma and 0.5 μg/ml for urine.     

Mathematical models for mixing in deep jet bioreactors: analysis

A mathematical model for single and multi step deep-jet bioreactors is presented. A stagewise approach based on macroscopic mechanistic model which divides the reactor into compartments with good quality of mixing and plug flow regions (macromixer), was used. For the mathematical representation of this model a system of differential equations, describing the concentration of tracer in structural elements based on mass balance, and the Runge-Kutta-Fehlberg numerical method of integration, was applied. The mixing time in a 300 dm³ tank was determined by conductivity method with NaCl as tracer.List of Symbols V _g dm³ total volume of liquid - V ₁; V ₆ dm³ volumes of ideally mixed compartments in the vessel - V ₂; V ₇ dm³ volumes of macromixer in the inner circulation flows - V ₃; V ₉ dm³ volumes of liquid phase in the pump - V ₄; V ₈ dm³ volumes of liquid phase in the pipe between the vessel and the pump - V ₅; V ₁₀ dm³ volumes of liquid phase in pipes between the pump and the air input system, including falling jet - F _E; F _E,1; F _E,2 dm³/s the inner volumetric circulation flow rates accross the macromixers - F _E,3; F _E,4 dm³/s exchanges volumetric flow rates between two ideally mixed compartments in the vessel - F _cir; F _1,cir; F _2,cir dm³/s external volumetric circulation flow rates (pumping capacity) - t _A s time interval of puls application - t _AA s time point of impuls application related to the free chosen point of simulation - t _end s end time of simulation - F _qu g²/dm⁶ sum of quadratic error - C _*,* kg/m³ concentration of the tracer in the indicated compartment - C ₀ kg/m³ concentration of the tracer before the injection - C _t kg/m³ concentration of the tracer at the indicated time - C kg/m³ theoretical concentration of full mixed tracer - i index of an arbitrary tank - C _sim kg/m³ calculated concentration of the tracer by numerical integration method     

Accelerated volatilization rates of selenium from different soils

Selenium (Se), an element found naturally in a variety of soils, can accumulate in drainage water of lands under intensive irrigation, even reaching levels that are toxic to mammals and birds. Volatilization of Se by soil microorganisms into dimethylselenide (DMSe) can be enhanced by certain soil amendments and, thus, be used as a soil remediation process. In an 8-wk laboratory study, five soils from California and one from Germany were spiked with⁷⁵SeO₃ ^2- (22.3 mg/kg Se). Two amino acids (DL-homocysteine and L-methionine), a carbohydrate (pectin), and a protein (zein) were tested as soil amendments. Gaseous⁷⁵Se emissions were trapped with activated carbon and measured in a gamma counter. Depending on soil type, the cumulative volatilization from the control flasks varied between 1.2% and 9.0% of applied⁷⁵Se. Both zein and L-methionine strongly increased volatilization (max. 43% of⁷⁵Se applied), whereas DL-homocysteine had a much smaller stimulating effect. Pectin showed a moderate effect, but enhanced Se volatilization rates were sustained much longer when compared to the zein amendment. Volatilization rates of Se followed a simple first-order reaction. Gaseous Se emission in the soils treated with L-methionine yielded an S-shaped curve, which fit a growth-modified first-order rate model. Although zein and L-methionine were the most favorable treatments enhancing Se volatilization, all six soils responded differently to the soil amendments.     

ROS up-regulation mediates Ras-induced changes of cell morphology and motility

Expression of activated Ras causes an increase in intracellular content of reactive oxygen species (ROS). To determine the role of ROS up-regulation in mediation of Ras-induced morphological transformation and increased cell motility, we studied the effects of hydrogen peroxide and antioxidant NAC on morphology of REF52 rat fibroblasts and their ability to migrate into the wound in vitro. Treatment with low dosages of hydrogen peroxide leading to 1.5- to 2-fold increase in intracellular ROS levels induced changes of cell shape, actin cytoskeleton organization, cell adhesions and migration resembling those in Ras-transformed cells. On the other hand, treatment with NAC attenuating ROS up-regulation in cells with conditional or constitutive expression of activated Ras led to partial reversion of morphological transformation and decreased cell motility. The effect of ROS on cell morphology and motility probably results from modulation of activity of Rac1, Rho, and cofilin proteins playing a key role in regulation of actin dynamics. The obtained data are consistent with the idea that ROS up-regulation mediates two key events in Ras-induced morphological transformation and cell motility: it is responsible for Rac1 activation and is necessary (though insufficient) for realization of Ras-induced cofilin dephosphorylation.     

Daily and circadian rhythms of locomotor activity in the American lobster, Homarus americanus

It is widely accepted that American lobsters, Homarus americanus (Milne-Edwards), are nocturnally active. However, the degree to which this rhythm is expressed by different individuals and the underlying causes of lobster activity rhythms, are poorly understood. In order to address these issues we recorded daily patterns of lobster locomotion using two novel techniques. In the first, reed switch assemblies were used to monitor the distance traveled by freely moving lobsters (n=43), each fitted with a small magnet, as they walked around a 1 m diameter racetrack. The advantages of this technique included: (1) lobsters were freely moving; (2) the system could be deployed in laboratory tanks or in the field and; (3) actual distances moved were measured, not just relative activity. The second technique involved placing individual lobsters (n=10) into custom-designed running wheels. This allowed for continuous monitoring of locomotor activity for extended durations (>45 days) under normal light/dark (L/D) cycles, as well as in constant darkness (D/D) and constant light (L/L).Under ambient light conditions lobsters in the racetracks moved an average of 60.1±6.5 m/day in flow-through seawater tanks. Overall, lobsters were significantly more active at night, moving 4.1±0.4 m/h in the dark vs. 1.0±0.2 m/h in the light. However, many of the lobsters moved as much during the day as during the night.Lobsters in the running wheels moved an average of 36.6±11.7 m/day and 80% expressed clear daily rhythms of activity, with a mean periodicity of 24.0±0.1 h under L/D conditions. Under D/D conditions 90% of the animals expressed free-running circadian rhythms with a mean periodicity of 24.2±0.3 h, indicating that this species possesses endogenous rhythmicity. While the running wheel results show that the nocturnal pattern of locomotor activity for this species is strongly influenced by an endogenous circadian clock, the results from the racetracks show that there is remarkable variability in the extent to which they express this pattern under natural conditions.     

Response of sorghum to a water gradient and potassium variable

Summary A field study utilizing a modified line-source sprinkler system was used to investigate the hydrologic balance of a sorghum crop subjected to water deficit stress. An additional objective was to evaluate the evapotranspiration-yield relationship and to determine if the water use efficiency could be improved through the addition of increased amounts of potassium. Sixteen lysimeters packed with a Greenfield sandy loam and planted to sorghum (Sorghum bicolor) were placed under a moisture gradient irrigation system. Four levels of potassium were used in the experiment (0, 100, 200 or 300 Kg/ha). The results indicated that when the leaching fraction was set at 0.15 near the source, the predicted leaching fraction would go to zero at 10 meters from the source. Although yield was related to evapotranspiration, a better fit to the yield data was obtained when a parameter that included an availability and deficit term was used. The relationship was not linear but instead was described by an exponential fit. This parameter was also highly sensitive to leaf water potential and diffusion resistance. Finally, water use efficiency increased with yield but showed no correlation with potassium application.     

Phylogenetic and biogeographical inferences for Pancratium (Amaryllidaceae), with an emphasis on the Mediterranean species based on plastid sequence data

The phylogenetics and biogeography of Pancratium (Amaryllidaceae) were investigated, with a focus on the Mediterranean and adjacent areas, with the aim of contributing new information towards a better understanding of the evolutionary history of the genus and the taxonomic placement of P. linosae and P. hirtum. To address these questions, we sequenced four plastid DNA markers: the ndhF and rbcL genes, the trnL^(UAA)–trnF^(GAA) intergenic spacer and the trnL^(UAA) intron, analysing them using parsimony, likelihood and Bayesian approaches. The results show that the relationships among the majority of the species are resolved; however, the relationships of one of the major clades of the genus are unresolved compared with the others. The phylogenetic and the dispersal–vicariance analyses show that Pancratium appears as a well‐structured group with interesting patterns of speciation. Notably, P. arabicum and P. linosae fall within the P. maritimum complex. In addition, P. hirtum is identical, in terms of plastid DNA sequences, to the P. trianthum accessions. The results provide new insights and help to formulate new working hypotheses for evolutionary biology of the genus. © 2012 The Linnean Society of London, Botanical Journal of the Linnean Society, 2012, 170 , 12–28.