Gene Expression Analysis Reveals Inhibition of Radiation-Induced TGFβ-Signaling by Hyperbaric Oxygen Therapy in Mouse Salivary Glands

A side effect of radiation therapy in the head and neck region is injury to surrounding healthy tissues such as irreversible impaired function of the salivary glands. Hyperbaric oxygen therapy (HBOT) is clinically used to treat radiation-induced damage but its mechanism of action is largely unknown. In this study, we investigated the molecular pathways that are affected by HBOT in mouse salivary glands two weeks after radiation therapy by microarray analysis. Interestingly, HBOT led to significant attenuation of the radiation-induced expression of a set of genes and upstream regulators that are involved in processes such as fibrosis and tissue regeneration. Our data suggest that the TGFβ-pathway, which is involved in radiation-induced fibrosis and chronic loss of function after radiation therapy, is affected by HBOT. On the longer term, HBOT reduced the expression of the fibrosis-associated factor α-smooth muscle actin in irradiated salivary glands. This study highlights the potential of HBOT to inhibit the TGFβ-pathway in irradiated salivary glands and to restrain consequential radiation induced tissue injury.     

Molecular phylogeny of the Valvatacea (Asteroidea: Echinodermata)

The Valvatacea is one the most ecologically important, taxonomically diverse, and widespread groups of post‐Palaeozoic (i.e. modern) Asteroidea. Classification within the group has been historically problematic. We present a comprehensively sampled, three‐gene (12S, 16S, early‐stage histone H3) molecular phylogenetic analysis of the Valvatacea. We include five of the six families within the Paxillosida, the monotypic Notomyotida, and 13 of the 16 families of the living Valvatida. The Solasteridae is removed from the Velatida (Spinulosacea) and joins the Ganeriidae and the Leilasteridae as members of the clade containing the Asterinidae. The Poraniidae is supported as the sister group to the large cluster of Valvatacea. Asteropseids and poraniids are phylogenetically distant, contrary to morphological evidence. Several goniasterid‐like ophidiasterids, such as Fromia and Neoferdina are supported as derived goniasterids rather than as Ophidiasteridae. The Benthopectinidae (Notomyotida) are supported as members of the Paxillosida as are two members of the Pseudarchasterinae that have traditionally been considered members of the Goniasteridae. Our data suggest that Antarctic valvataceans may be derived from sister taxa in adjacent regions. © 2011 The Linnean Society of London, Zoological Journal of the Linnean Society, 2011, 161 , 769–788.     

Factors Influencing the in vitro Culture of the Rumen Ciliate Ophryoscolex purkynei Stein

SYNOPSIS. A method for the in vitro culture of O. purkynei was developed which permitted the continuous culture of this protozoon in clone culture for a. period of 32 months. The shortest division time was 24 hr. The concentration of cells varied between 700 and 1000/ml in the routine procedure. Variations in spination which occurred in the clone culture suggested that this characteristic was of doubtful taxonomic importance.
Ground wheat and alfalfa served as substrates but soluble sugars did not. Green plant material appeared to be necessary for continued growth of the protozoa. Ingestion of a large streptococcus was demonstrated by vital staining of a mixed population of bacteria with tetrazolium prior to incubation with the protozoan suspension in the presence of wheat.
O. purkynei can tolerate exposure to variations in osmotic pressure, temperature, and oxygen which are consonant with its transfer in nature by grooming or ingestion of contaminated food or drink.     

Delayed setting of the photoperiodic response in recombinant clones of the aphid species Sitobion avenae

1. To explore the possible causes of apparent changes in reproductive mode from obligate to cyclical parthenogenesis over time in recombinant clones of the aphid Sitobion avenae Fabricius, all F1 progenies from various crosses were tested for several consecutive years for sexual morph production, after several weeks' exposure to a short photoperiod. 2. Variable proportions of the F1 progenies from selfing and outcrossing holocyclic clones did not produce mating females when induction was attempted in the year of hatching, but only after further induction, the following year or after. This ‘delayed setting of the photoperiodic response’ (DSPR) was much stronger in recombinants from crosses involving only clones from oceanic regions than in those involving one clone from a region with a continental climate. 3. F1 progenies resulting from crosses between one holocyclic and one intermediate clone did not show DSPR. It appeared again in the F2. 4. DSPR preferentially affected the latest hatched clones in a given progeny. 5. This phenomenon is neither an experimental artefact nor as a result of clone contamination. It appears to be because of a genetically controlled quantitative trait affecting the length of the ‘interval timer’, and may represent an adaptation of holocyclic aphid clones from oceanic regions to unpredictable winter climates.     

Development of a monoclonal antibody against recombinant neuroendocrine 7B2 protein

Mouse monoclonal antibody MON-100 was raised against the neuroendocrine protein 7B2 using bacterially produced hybrid proteins. In Western blot analysis, MON-100 reacted with 3 different 7B2 hybrid proteins and not with the respective carrier proteins. Furthermore, MON-100 was reactive with recombinant 7B2 cleaved from a hybrid protein. In an immunohistochemical study, MON-100 exhibited strong reactivity with the intermediate lobe of the Xenopus pituitary gland, a tissue previously shown to contain 7B2 mRNA. Using MON-100, immunoprecipitation analysis of newly synthesized proteins produced by in vitro incubated Xenopus neurointermediate lobes revealed the biosynthesis of a single protein of Mr 24 kDa, the expected size of the 7B2 protein. It appears, therefore, that the anti-7B2 monoclonal antibody MON-100 can be successfully used for Western blot analysis and immunohistochemical analysis as well as for immunoprecipitation experiments.     

Metabolic origin of δ15N values in nitrogenous compounds from Brassica napus L. leaves

Nitrogen isotope composition (δ¹⁵N) in plant organic matter is currently used as a natural tracer of nitrogen acquisition efficiency. However, the δ¹⁵N value of whole leaf material does not properly reflect the way in which N is assimilated because isotope fractionations along metabolic reactions may cause substantial differences among leaf compounds. In other words, any change in metabolic composition or allocation pattern may cause undesirable variability in leaf δ¹⁵N. Here, we investigated the δ¹⁵N in different leaf fractions and individual metabolites from rapeseed (Brassica napus) leaves. We show that there were substantial differences in δ¹⁵N between nitrogenous compounds (up to 30‰) and the content in (¹⁵N enriched) nitrate had a clear influence on leaf δ¹⁵N. Using a simple steady‐state model of day metabolism, we suggest that the δ¹⁵N value in major amino acids was mostly explained by isotope fractionation associated with isotope effects on enzyme‐catalysed reactions in primary nitrogen metabolism. δ¹⁵N values were further influenced by light versus dark conditions and the probable occurrence of alternative biosynthetic pathways. We conclude that both biochemical pathways (that fractionate between isotopes) and nitrogen sources (used for amino acid production) should be considered when interpreting the δ¹⁵N value of leaf nitrogenous compounds.