Angiopoietin-1/Tie-2 signal after focal traumatic brain injury is potentiated by BQ788, an ETB receptor antagonist, in the mouse cerebrum: Involvement in recovery of blood-brain barrier function.

Angiopoietin-1, an angiogenic factor, stabilizes brain microvessels through Tie-2 receptor tyrosine kinase. In traumatic brain injury, blood-brain barrier (BBB) disruption is an aggravating factor that induces brain edema and neuroinflammation. We previously showed that BQ788, an endothelin ETB receptor antagonist, promoted recovery of BBB function after lateral fluid percussion injury (FPI) in mice. To clarify the mechanisms underlying BBB recovery mediated by BQ788, we examined the involvements of the angiopoietin-1/Tie-2 signal. When angiopoietin-1 production and Tie-2 phosphorylation were assayed by quantitative reverse transcription polymerase chain reaction and western blotting, increased angiopoietin-1 production and Tie-2 phosphorylation were observed in 7-10 days after FPI in the mouse cerebrum, whereas no significant effects were obtained at 5 days. When BQ788 (15 nmol/day, i.c.v.) were administered in 2-5 days after FPI, increased angiopoietin-1 production and Tie-2 phosphorylation were observed. Immunohistochemical observations showed that brain microvessels and astrocytes contained angiopoietin-1 after FPI, and brain microvessels also contained phosphorylated Tie-2. Treatment with endothelin-1 (100 nM) decreased angiopoietin-1 production in cultured astrocytes and the effect was inhibited by BQ788 (1 µM). Five days after FPI, increased extravasation of Evans blue dye accompanied by reduction in claudin-5, occludin, and zonula occludens-1 proteins were observed in mouse cerebrum while these effects of FPI were reduced by BQ788 and exogenous angiopoietin-1 (1 µg/day, i.c.v.). The effects of BQ788 were inhibited by co-administration of a Tie-2 kinase inhibitor (40 nmol/day, i.c.v.). These results suggest that BQ788 administration after traumatic brain injury promotes recovery of BBB function through activation of the angiopoietin-1/Tie-2 signal.

Statistical Modeling of Within-Laboratory Precision Using a Hierarchical Bayesian Approach.

BACKGROUND: Reproducibility has been well-studied in the field of food analysis; the relative standard deviation is said to follow the Horwitz curve with certain exceptions. However, little systematic research has been done on predicting repeatability or intermediate precision. OBJECTIVE: We developed a regression method to estimate within-laboratory standard deviations using hierarchical Bayesian modeling and analyzing duplicate measurement data obtained from actual laboratory tests. METHODS: The Hamiltonian Monte Carlo method was employed and implemented using R with Stan. The basic structure of the statistical model was assumed to be a chi-squared distribution, the fixed effect of the predictor was assumed to be a nonlinear function with a constant term and a concentration-dependent term, and the random effects were assumed to follow a lognormal distribution as a hierarchical prior. RESULTS: By analyzing over 300 instances, we obtained regression results that fit well with the assumed model, except for moisture, which was a method-defined analyte. The developed method applies to a wide variety of analytes measured using general principles, including spectroscopy, GC, and HPLC. Although the estimated precisions were within the HorRat(r) criteria, some cases using high-sensitivity detectors, such as mass spectrometers, showed standard deviations below that range. CONCLUSION: We propose utilizing the within-laboratory precision predicted by the model established in this study for internal quality control and measurement uncertainty estimation without considering the sample matrices. HIGHLIGHTS: Performing statistical modeling on data from double analysis, which is conducted as a part of internal quality controls, will simplify the estimation of the precision that fits each analytical system in a laboratory.

[Analytical method of volatile substances in styrene polymers by head-space gas chromatography using o-dichlorobenzene].

A quantitative analytical method for 8 volatile substances (such as styrene) in styrene polymers that are not soluble in tetrahydrofuran (THF) and dimethylformamide was developed. The sample was chopped finely and a 0.1 g portion was weighed in a head-space vial. To this was added 2 mL of o-dichlorobenzene (DCB) including 50 µg/mL of internal standard. The vial was sealed and heated at 140℃ for 1 hour and then 1 mL of the head-space gas was injected into a gas chromatograph using an automatic sampler. Samples of syndiotactic polystyrene, styrene block copolymer and modified polyphenylenether were dissolved or dispersed in DCB. Separation and accuracy of the method were satisfactory. Recoveries were 95-113% at the spiked concentration of 300 µg/g. The method was confirmed to be suitable for general styrene polymers that are soluble in THF. Equivalent results were obtained with this method and the method in "Specifications and Standards for Food, Food Additives, etc. " for general purpose polystyrene, high impact polystyrene, styrene/acrylonitrile resin and acrylonitrile/butadiene/styrene resin.

Exploitation or cooperation? Evolution of a host (ciliate)-benefiting alga in a long-term experimental microcosm culture.

Controversy persists as to whether the acquisition of beneficial metabolic functions via endosymbiosis can occur suddenly on an evolutionary time scale. In this study, an early stage of endosymbiotic associations, which evolved from previously unassociated auto (photo)- and heterotrophic unicellular organisms was analyzed using an experimental ecosystem model, called CET microcosm. This ecosystem model was composed of a green alga (Micractinium sp.; formerly described as Chlorella vulgaris), a bacterium (Escherichia coli), and a ciliate (Tetrahymena thermophila). Our previous study using a CET microcosm that was cultured 3-5 years revealed that fitness of the ciliate increased by harboring algal cells within its own cells. This fact suggested three possibilities: (i) the ciliate evolved the ability to exploit intracellular algal cells ("exploiter ciliate hypothesis"), (ii) the alga evolved the ability to benefit the host ciliate by providing photosynthates ("cooperator alga hypothesis"), and (iii) a combination of (i) and (ii). To test these hypotheses, two-by-two co-cultures were conducted between the ancestral or derived ciliate and the ancestral or derived alga. The experimental results demonstrated that a cooperative alga evolved in the microcosm, although the possibility remains that an exploitative genotype of the ciliate might also exist in the population as a polymorphism. Remarkably, an algal isolate prolonged the longevity of not only the isolated ciliate, but also the ancestral ciliate. This result suggests that once a cooperative algal genotype evolves in a local population, it can then be transmitted to other individuals of the prospective host species and spread rapidly beyond the local range due to its positive effect on the host fitness. Such transmission suggests the possibility of a sudden acquisition of beneficial autotrophic function by the pre-associated host.

Thiosulfate oxidation by a moderately thermophilic hydrogen-oxidizing bacterium, Hydrogenophilus thermoluteolus.

The moderately thermophilic Betaproteobacterium, Hydrogenophilus thermoluteolus, not only oxidizes hydrogen, the principal electron donor for growth, but also sulfur compounds including thiosulfate, a process enabled by sox genes. A periplasmic extract of H. thermoluteolus showed significant thiosulfate oxidation activity. Ten genes apparently involved in thiosulfate oxidation (soxEFCDYZAXBH) were found on a 9.7-kb DNA fragment of the H. thermoluteolus chromosome. The proteins SoxAX, which represent c-type cytochromes, were co-purified from the cells of H. thermoluteolus; they enhanced the thiosulfate oxidation activity of the periplasmic extract when added to the latter.