Effect of bradykinin analogues on the B1 receptor of rat ileum

The longitudinal muscle of isolated rat ileum is a sensitive bioassay suitable for testing compounds with antagonistic effects on the B(1) receptor. Bradykinin analogues with replacement of proline by alkyl-substituted phenylalanine at position 7 are effective on this receptor as entire molecules and have a stronger antagonistic effect than on the B(2) receptor. A corresponding desArg(9)-compound has a specific effect on the B(1) receptor and a very high antagonistic potency. [LNMPhe(2)]bradykinin as a compound without any replacement at position 7 or 8 shows antagonistic activity as well.     

Malaria and World War II: German malaria experiments 1939-45

The epidemiological and pharmacological fight against malaria and German malaria research during the Nazi dictatorship were completely under the spell of war. The Oberkommando des Heeres (German supreme command of the army) suffered the bitter experience of unexpected high losses caused by malaria especially at the Greek front (Metaxes line) but also in southern Russia and in the Ukraine. Hastily raised anti-malaria units tried to teach soldiers how to use the synthetic malaria drugs (Plasmochine, Atebrine) properly. Overdoses of these drugs were numerous during the first half of the war whereas in the second half it soon became clear that it would not be possible to support the army due to insufficient quantities of plasmochine and atebrine. During both running fights and troop withdrawals at all southern and southeastern fronts there was hardly any malaria prophylaxis or treatment. After war and captivity many soldiers returned home to endure heavy malaria attacks. In German industrial (Bayer, IG-Farben) and military malaria laboratories of the Heeres-Sanit?ts-Akademie (Army Medical Academy) the situation was characterised by a hasty search for proper dosages of anti-malaria drugs, adequate mechanical and chemical prophylaxis (Petroleum, DDT, and other insecticides) as well as an anti-malaria vaccine. Most importantly, large scale research for proper atebrine and plasmochine dosages was conducted in German concentration camps and mental homes. In Dachau Professor Claus Schilling tested synthetic malaria drugs and injected helpless prisoners with high and sometimes lethal doses. Since the 1920s he had been furiously looking for an anti-malaria vaccine in Italian mental homes and from 1939 he continued his experiments in Dachau. Similar experiments were also performed in Buchenwald and in a psychiatric clinic in Thuringia, where Professor Gerhard Rose tested malaria drugs with mentally ill Russian prisoners of war. Schilling was put to death for his criminal research in 1946, Rose was condemned to lifelong imprisonment in 1947, though, not for his malaria research but for his dreadful experiments with epidemic typhus sera which he also had performed in concentration camps and with prisoners of war in Russia.     

Bassoon,a Novel Zinc-finger CAG/Glutamine-repeat Protein Selectively Localized at the Active Zone of Presynaptic Nerve Terminals

The molecular architecture of the cytomatrix of presynaptic nerve terminals is poorly understood. Here we show that Bassoon, a novel protein of >400,000 M _r, is a new component of the presynaptic cytoskeleton. The murine bassoon gene maps to chromosome 9F. A comparison with the corresponding rat cDNA identified 10 exons within its protein-coding region. The Bassoon protein is predicted to contain two double-zinc fingers, several coiled-coil domains, and a stretch of polyglutamines (24 and 11 residues in rat and mouse, respectively). In some human proteins, e.g., Huntingtin, abnormal amplification of such poly-glutamine regions causes late-onset neurodegeneration. Bassoon is highly enriched in synaptic protein preparations. In cultured hippocampal neurons, Bassoon colocalizes with the synaptic vesicle protein synaptophysin and Piccolo, a presynaptic cytomatrix component. At the ultrastructural level, Bassoon is detected in axon terminals of hippocampal neurons where it is highly concentrated in the vicinity of the active zone. Immunogold labeling of synaptosomes revealed that Bassoon is associated with material interspersed between clear synaptic vesicles, and biochemical studies suggest a tight association with cytoskeletal structures. These data indicate that Bassoon is a strong candidate to be involved in cytomatrix organization at the site of neurotransmitter release.     

Somatodendritic serotonin release and re-uptake in mouse embryonic stem cell-derived serotonergic neurons

Serotonergic neurotransmission plays an important role during neural development. Serotonergic dysfunction is observed in various psychiatric disorders and many psychoactive drugs target proteins on serotonergic neurons. Serotonergic neurons are located in the raphé nuclei and densely innervate the whole brain. The low number and the intricate accessibility of these neurons do not allow to culture them and therefore to date it was impossible to study drug-target interactions on bona fide serotonergic neurons. In order to circumvent such problems we have developed a protocol that allows the rapid and efficient generation of serotonergic neurons from mouse embryonic stem cells. Neuronal precursors were obtained by neuronal stem sphere formation in floating culture in the presence of various mitogens. Differentiation into neurons was induced by withdrawal of the mitogens. About 90% of the resulting neurons exhibited a serotonergic phenotype as judged by immunostaining against serotonin, its synthesising enzyme tryptophan hydroxylase 2, the serotonin transporter as well as 5-HT1(A) and 5-HT1(B) autoreceptors. In addition, we found expression of the vesicular monoamine transporter vMAT2 and the presynaptic protein Bassoon, which is involved in organizing the assembly of the presynaptic active zone. Depolarisation-induced calcium influx was visualised by Fluo-4, and accompanying exocytotic events by FM dye staining. Proteins involved in 5-HT release and re-uptake as well as depolarisation evoked exocytosis were evenly co-distributed on neurites and cell bodies suggesting that ES cell-derived serotonergic neurons also exhibit somatodendritic release comparable to serotonergic neurons in the raphé nuclei.     

Identification of NoxD/Pro41 as the homologue of the p22phox NADPH oxidase subunit in fungi

NADPH oxidases (Nox) are membrane complexes that produce O₂^?. Researches in mammals, plants and fungi highlight the involvement of Nox‐generated ROS in cell proliferation, differentiation and defense. In mammals, the core enzyme gp91^phox/Nox2 is associated with p22^phox forming the flavocytochrome b₅₅₈ ready for activation by a cytosolic complex. Intriguingly, no homologue of the p22^phox gene has been found in fungal genomes, questioning how the flavoenzyme forms. Using whole genome sequencing combined with phylogenetic analysis and structural studies, we identify the fungal p22^phox homologue as being mutated in the Podospora anserina mutant IDC⁵⁰⁹. Functional studies show that the fungal p22^phox, PaNoxD, acts along PaNox1, but not PaNox2, a second fungal gp91^phox homologue. Finally, cytological analysis of functional tagged versions of PaNox1, PaNoxD and PaNoxR shows clear co‐localization of PaNoxD and PaNox1 and unravel a dynamic assembly of the complex in the endoplasmic reticulum and in the vacuolar system.     

Correlating SHAPE signatures with three-dimensional RNA structures

Selective 2'-hydroxyl acylation analyzed by primer extension (SHAPE) is a facile technique for quantitative analysis of RNA secondary structure. In general, low SHAPE signal values indicate Watson-Crick base-pairing, and high values indicate positions that are single-stranded within the RNA structure. However, the relationship of SHAPE signals to structural properties such as non-Watson-Crick base-pairing or stacking has thus far not been thoroughly investigated. Here, we present results of SHAPE experiments performed on several RNAs with published three-dimensional structures. This strategy allows us to analyze the results in terms of correlations between chemical reactivities and structural properties of the respective nucleotide, such as different types of base-pairing, stacking, and phosphate-backbone interactions. We find that the RNA SHAPE signal is strongly correlated with cis-Watson-Crick/Watson-Crick base-pairing and is to a remarkable degree not dependent on other structural properties with the exception of stacking. We subsequently generated probabilistic models that estimate the likelihood that a residue with a given SHAPE score participates in base-pairing. We show that several models that take SHAPE scores of adjacent residues into account perform better in predicting base-pairing compared with individual SHAPE scores. This underscores the context sensitivity of SHAPE and provides a framework for an improved interpretation of the response of RNA to chemical modification.     

Inhibition of histone deacetylases in inflammatory bowel diseases

This review, comprised of our own data and that of others, provides a summary overview of histone deacetylase (HDAC) inhibition on intestinal inflammation as well as inflammation-mediated carcinogenesis. Experimental colitis in mice represents an excellent in vivo model to define the specific cell populations and target tissues modulated by inhibitors of HDAC. Oral administration of either suberyolanilide hydroxamic acid (SAHA) or ITF2357 results in an amelioration in these models, as indicated by a significantly reduced colitis disease score and histological score. This effect was paralleled by suppression of proinflammatory cytokines at the site of inflammation as well as specific changes in the composition of cells within the lamina propria. In addition, tumor number and size was significantly reduced in two models of inflammation-driven tumorigenesis, namely interleukin (IL)-10-deficient mice and the azoxymethane-dextran sulfate sodium (DSS) model, respectively. The mechanisms affected by HDAC inhibition, contributing to this antiinflammatory and antiproliferative potency will be discussed in detail. Furthermore, with regard to the relevance in human inflammatory bowel disease, the doses of ITF2357 considered safe in humans and the corresponding serum concentrations are consistent with the efficacious dosing used in our in vivo as well as in vitro experiments. Thus, the data strongly suggest that HDAC inhibitors could serve as a therapeutic option in inflammatory bowel disease.     

The presence of a paternal grandmother lengthens interbirth interval following the birth of a granddaughter in Krummhörn (18th and 19th centuries)

Because only daughters inherit the paternal X-chromosome, an asymmetry in adaptive investment decisions has been suggested for certain patrilineal kin. Namely, paternal grandmothers (PGMs) may favor a granddaughter over a grandson, because (within the limits of paternity uncertainty) the former definitely carries one of their X-chromosomes, while the latter definitely does not. Here, we test the hypothesis that the PGMs' sex-specific favoritism influences reproductive scheduling. Using family-reconstitution data, we analyzed interbirth intervals (IBIs) in the historical population from the Krummhörn (Ostfriesland, Germany). In order to account for potentially timevarying effects on IBIs we applied (and combined) both the additive hazards regression of Aalen and the Cox proportional hazards model. We found that the presence of the PGM but not that of the maternal grandmother (MGM), correlates with the IBI following the birth of a grandchild as a function of the grandchild's sex. Specifically, in the presence of a PGM, the IBIs following the birth of a granddaughter are longer than in her absence. However, contrary to predictions from theoretical life history framework, model estimates for a PGM's effect on a mother's IBI did not significantly vary over time This study supports the hypothesis that PGM behavior differs according to her grandchild's sex. Further research should now explore the biological mechanism underlying this phenomenon.