Lifetime Dependent Variation of Stress Hormone Metabolites in Feces of Two Laboratory Mouse Strains

Non-invasive measurement of stress hormone metabolites in feces has become routine practice for the evaluation of distress and pain in animal experiments. Since metabolism and excretion of glucocorticoids may be variable, awareness and adequate consideration of influencing factors are essential for accurate monitoring of adrenocortical activity. Reference values are usually provided by baselines compiled prior to the experiment and by age matched controls. The comparison of stress hormone levels between animals of different ages or between studies looking at hormone levels at the beginning and at the end of a long term study might be biased by age-related effects. In this study we analyzed fecal corticosterone metabolites (FCM) during the lifetime of untreated female mice of the strains C57BL/6NCrl and Crl:CD1. For this purpose feces for each individual mouse were collected every two months over a period of 24 hours, at intervals of four hours, until the age of 26 months. Results of the study revealed that age of the animals had a significant impact on the level and circadian rhythm of stress hormone metabolites. Furthermore, long-term observation of mice revealed a strain specific excretion profile of FCM influenced by strong seasonal variability.     

Global expression analysis of nucleotide binding site-leucine rich repeat-encoding and related genes in Arabidopsis

Background  

Nucleotide binding site-leucine rich repeat (NBS-LRR)-encoding genes comprise the largest class of plant disease resistance genes. The 149 NBS-LRR-encoding genes and the 58 related genes that do not encode LRRs represent approximately 0.8% of all ORFs so far annotated in Arabidopsis ecotype Col-0. Despite their prevalence in the genome and functional importance, there was little information regarding expression of these genes.     

Characterization of dentin matrix protein 1 gene in crocodilia.

Several clones containing DMP1 cDNA were isolated from a caiman tooth library by screening with a platypus DMP1 probe. The caiman DMP1 shows little amino acid sequence similarity to mammalian DMP1s for much of its length. A few highly conserved regions can, however, be identified that correspond to the slowly evolving parts of the corresponding mammalian genes. Southern blot analysis using probes comprising either conserved regions or longer segments of the gene indicates that only a single DMP1 locus exists. In coding regions, exon-intron boundaries and reading frames are shared by caiman and mammalian genes with the exception of exons 1 and 5, which are longer in the caiman. The repetitive sequence of the last exon is shared by mammals and caiman as are the high Ser content and acidity due to a high proportion of Asp and Glu residues. The conserved mammalian cell-attachment signal Arg-Gly-Asp is absent in the caiman DMP1. In contrast to the amelogenin gene, the DMP1 gene appears to evolve rapidly in vertebrates.     

TatA and TatB generate a hydrophobic mismatch important for the function and assembly of the Tat translocon in Escherichia coli

The twin-arginine translocation (Tat) system serves to translocate folded proteins across energy-transducing membranes in bacteria, archaea, plastids, and some mitochondria. In Escherichia coli, TatA, TatB, and TatC constitute functional translocons. TatA and TatB both possess an N-terminal transmembrane helix (TMH) followed by an amphipathic helix. The TMHs of TatA and TatB generate a hydrophobic mismatch with the membrane, as the helices comprise only 12 consecutive hydrophobic residues; however, the purpose of this mismatch is unclear. Here, we shortened or extended this stretch of hydrophobic residues in either TatA, TatB, or both and analyzed effects on translocon function and assembly. We found the WT length helices functioned best, but some variation was clearly tolerated. Defects in function were exacerbated by simultaneous mutations in TatA and TatB, indicating partial compensation of mutations in each by the other. Furthermore, length variation in TatB destabilized TatBC-containing complexes, revealing that the 12-residue-length is important but not essential for this interaction and translocon assembly. To also address potential effects of helix length on TatA interactions, we characterized these interactions by molecular dynamics simulations, after having characterized the TatA assemblies by metal-tagging transmission electron microscopy. In these simulations, we found that interacting short TMHs of larger TatA assemblies were thinning the membrane and—together with laterally-aligned tilted amphipathic helices—generated a deep V-shaped membrane groove. We propose the 12 consecutive hydrophobic residues may thus serve to destabilize the membrane during Tat transport, and their conservation could represent a delicate compromise between functionality and minimization of proton leakage.     

Response of moist-air receptor on antenna of the stick insect,Carausius morosus,to step changes in temperature

Summary The moist-air sensory cell in the antennal mound-shaped sensillum of Carausius responds to changes in relative humidity brought about by changing either the temperature (T) of the air or the partial pressure of water vapor (Pw) (Figs. 1, 5). When changes in either parameter cause relative humidity (Hr) to rise between roughly 5% and 55%, the same Hr-changes elicit very close to the same responses, no matter how Hr is changed (Figs. 2, 3). Even the resolving power for upward Hr-steps produced by lowering T is very close to that produced by raising Pw: 7.4% vs. 6.3%. Thus upward transients in impulse frequency (F) may be read off directly as quantitative rises in Hr. Whether the rise in Hr as signalled by the moist-air cell is the result of a rise in Pw or of a drop in T could well be indicated by the simultaneous reactions of the cold and dry-air cells (Figs. 1, 5). The moist-air cell also reacts to downward step changes in Hr, but very differently, depending on how they are brought about. Upward steps in T have a much larger effect on F than corresponding downward steps in Pw (Fig. 4). This result demonstrates that changes in relative humidity do not suffice to explain the changes in the activity of the moist-air cell. The receptor may be better construed as bimodal, reacting to changes in T and Pw with independent sensitivities and well matched to relative humidity when changes in either lead to increases in this parameter. The moistair cell's responses to T-steps at Pw = 0 (Figs. 1, 5) offer some support for this interpretation.Abbreviations F impulse frequency in impulses/s (imp/s) - Hr relative humidity in % - Ps saturation pressure of water vapor in torr - Pw partial pressure of water vapor in torr - r correlation coefficient - T temperature in °C     

Cold-receptor cells supply both cold- and warm-responsive projection neurons in the antennal lobe of the cockroach

The temperature receptor cells on the cockroach antennae are all excited by rapid cooling. In the antennal lobe, however, cold- as well as warm-responsive neurons occur. They are excited either by rapid step-like cooling or rapid step-like warming. Responses to such temperature transients do not show, however, whether antennal lobe neurons convey information on slowly changing temperatures typical of temperature gradients used for orientation. In contrast slow temperature changes permit an analysis of the effects of both instantaneous temperature and its rate of change. We compared the effect of slow temperature oscillations on the responses of antennal cold-receptors cells and cold- and warm-responsive projection neurons. In all cases the discharge rates were modulated by the temperature oscillations. They displayed a double dependence on instantaneous temperature and its rate of change. Information about cooling and warming, first contained in the output of a single cold-receptor cell diverges to form the parallel pathways of cold- and warm-responsive projection neurons, thereby in particular improving the detection of fluctuations in temperature.     

Immunohistochemical Assessment of Phosphorylated mTORC1-Pathway Proteins in Human Brain Tumors

BackgroundCurrent pathological diagnostics include the analysis of (epi-)genetic alterations as well as oncogenic pathways. Deregulated mammalian target of rapamycin complex 1 (mTORC1) signaling has been implicated in a variety of cancers including malignant gliomas and is considered a promising target in cancer treatment. Monitoring of mTORC1 activity before and during inhibitor therapy is essential. The aim of our study is to provide a recommendation and report on pitfalls in the use of phospho-specific antibodies against mTORC1-targets phospho-RPS6 (Ser235/236; Ser240/244) and phospho-4EBP1 (Thr37/46) in formalin fixed, paraffin embedded material.ConclusionHere we provide a recommendation for phospho-specific immunohistochemistry for patient-orientated therapy decisions and monitoring treatment response.     

Genotype × environment interaction in the allometry of body,genitalia and signal traits in Enchenopa treehoppers (Hemiptera: Membracidae)

Developmental plasticity may promote divergence by exposing genetic variation to selection in novel ways in new environments. We tested for this effect in the static allometry (i.e. scaling on body size) of traits in advertisement signals, body and genitalia. We used a member of the Enchenopa binotata species complex of treehoppers – a clade of plant‐feeding insects in which speciation is associated with colonization of novel environments involving marked divergence in signals, subtle divergence in body size and shape, and no apparent divergence in genitalia. We found no change in mean allometric slopes across environments, but substantial genetic variation and genotype × environment interaction (G × E) in allometry. The allometry of signal traits showed the most genetic variation and G × E, and that of genitalia showed the weakest G × E. Our findings suggest that colonizing novel environments may have stronger diversifying consequences for signal allometry than for genitalia allometry. © 2011 The Linnean Society of London, Biological Journal of the Linnean Society, 2012, 105 , 187–196.