Alterations of alkaline phosphatase activity during adaptation of Escherichia coli to phosphite and hypophosphite

When Escherichia coli cells were grown in media containing either phosphite or hypophosphite as the sole source of phosphorus, they responded to this situation primarily in the same way as phosphatelimited cultures: The activity of alkaline phosphatase increased drastically, which under natural conditions would enable the cells to compklensatae for the shortage increased drastically, which under natural conditions would enable the cells to compensate for the shortage of phosphate. Subsequent transfers, however, resulted in a quite different response: While the phosphatase activity of phosphate-limited cells stays at a high derepressed level, its increase was followed by a gradual decline in organisms grown on phosphite or hypophosphite. After eight to ten transfers on these P-compounds, phosphatase activity was back to its initial, repressed, low level, indicating that the cells were fully adapted to these substrates. Adaptation to either PO ₃ ^3- or PO ₂ ^3- was completely abolished if the cells were again grown with PO ₄ ^3- as P-source, whereafter the entire process of adaptation had to be repeated. The observed adaptation pattern, reflected by the alterations of phosphatase activity, was qualitatively equal with PO ₃ ^3- and PO ₂ ^3- , but quantitatively different, because the response to hypophosphite gave much higher values than the increase obtained with phosphite.Phosphite-adapted cells are not simultaneously adapted to hypophosphite, but their response to the latter was less intense than observed after direct transfers from PO ₄ ^3- to PO ₂ ^3- . Adaptation to hypophosphite, however, led simultaneously to phosphite adaptation, so that these cells can utilize both P-compounds as a substitute for phosphate.     

Dietary inorganic nitrate mobilizes circulating angiogenic cells

Nitric oxide (NO) was implicated in the regulation of mobilization and function of circulating angiogenic cells (CACs). The supposedly inert anion nitrate, abundant in vegetables, can be stepwise reduced in vivo to form nitrite, and consecutively NO, representing an alternative to endogenous NO formation by NO synthases. This study investigated whether inorganic dietary nitrate influences mobilization of CACs. In a randomized double-blind fashion, healthy volunteers ingested 150 ml water with 0.15 mmol/kg (12.7 mg/kg) of sodium nitrate, an amount corresponding to 100-300 g of a nitrate-rich vegetable, or water alone as control. Mobilization of CACs was determined by the number of CD34(+)/KDR(+) and CD133(+)/KDR(+) cells using flow cytometry and the mobilization markers stem cell factor (SCF) and stromal cell-derived factor-1a (SDF-1α) were determined in plasma via ELISA. Nitrite and nitrate were measured using high-performance liquid chromatography and reductive gas-phase chemiluminescence, respectively. NOS-dependent vasodilation was measured as flow-mediated vasodilation. Further mechanistic studies were performed in mice after intravenous application of nitrite together with an NO scavenger to identify the role of nitrite and NO in CAC mobilization. Nitrate ingestion led to a rise in plasma nitrite together with an acute increase in CD34(+)/KDR(+) and CD133(+)/KDR(+)-CACs along with increased NOS-dependent vasodilation. This was paralleled by an increase in SCF and SDF-1α and the maximal increase in plasma nitrite correlated with CD133(+)/KDR(+)-CACs (r=0.73, P=0.016). In mice, nitrate given per gavage and direct intravenous injection of nitrite led to CAC mobilization, which was abolished by the NO scavenger cPTIO, suggesting that nitrite mediated its effect via formation of NO. Dietary inorganic nitrate acutely mobilizes CACs via serial reduction to nitrite and NO. The nitrate-nitrite-NO pathway could offer a novel nutritional approach for regulation of vascular regenerative processes.     

EVOLUTION OF MALE COLORATION DURING A POST‐PLEISTOCENE RADIATION OF BAHAMAS MOSQUITOFISH (GAMBUSIA HUBBSI)

Sexual signal evolution can be complex because multiple factors influence the production, transmission, and reception of sexual signals, as well as receivers’ responses to them. To grasp the relative importance of these factors in generating signal diversity, we must simultaneously investigate multiple selective agents and signaling traits within a natural system. We use the model system of the radiation of Bahamas mosquitofish (Gambusia hubbsi) inhabiting blue holes to test the effects of resource availability, male body size and other life‐history traits, key aspects of the transmission environment, sex ratio, and predation risk on variation in multiple male color traits. Consistent with previous work examining other traits in this system, several color traits have repeatedly diverged between predation regimes, exhibiting greater elaboration in the absence of predators. However, other factors proved influential as well, with variation in resource levels, body size, relative testes size, and background water color being especially important for several color traits. For one prominent signaling trait, orange dorsal fins, we further confirmed a genetic basis underlying population differences using a laboratory common‐garden experiment. We illustrate a promising approach for gaining a detailed understanding of the many contributing factors in the evolution of multivariate sexual signals.     

Wheat Germ Agglutinin Staining as a Suitable Method for Detection and Quantification of Fibrosis in Cardiac Tissue after Myocardial Infarction

The quantification of fibrotic tissue is an important task in the analysis of cardiac remodeling. The use of established fibrosis staining techniques is limited on frozen cardiac tissue sections due to a reduced color contrast compared to paraffin embedded sections. We therefore used FITC-labeled wheat germ agglutinin (WGA), which marks fibrotic tissue in comparable quality as the established picrosirius red (SR) staining, for the staining of post myocardial infarction scar tissue. The fibrosis amount was quantified in a histogram-based approach using the non-commercial image processing program ImageJ. Our results clearly demonstrate that WGA-FITC is a suitable marker for cardiac fibrosis in frozen tissue sections. In combination with the histogram-based analysis, this new quantification approach is i) easy and fast to perform; ii) suitable for raw frozen tissue sections; and iii) allows the use of additional antibodies in co-immunostaining.Key words: Cardiac fibrosis, myocardial infarction, wheat germ agglutinin, fibrosis quantification, histogram-based analysis     

Siliciumstoffwechsel bei Mikroorganismen

Summary Soil bacteria which have been used in earlier experiments to demonstrate an active uptake of silicon, loose phosphate during silicon uptake when cultured in P-free medium. This could be shown by comparable determinations of the phosphate and silicon concentration of the cells. Under the conditions given in our experiments the exchange of Si for P lies in the range of 1:2. By addition of rising P-concentrations to media with constant concentration of Si, it was shown that about 100 P/ml will completely inhibit the uptake of silicon within 24 hours. Increasing concentrations of phosphate going along with decreasing concentrations of silicate showed to cause a linear decrease of Si-uptake intensity within the first 24 hours in the range of 20–100 P/ml. Above these concentrations (and the proportion of Si/P=1:4) silicon uptake is completely inhibited independent of phosphate concentrations. About 10% of the silicon incorporated can be extracted from the cells with ethanol in the form of instable, easily hydrolysable complexes. The entire silicon of the cells is completely exchanged against phosphate when silicon containing cells are cultured in Si-free phosphate medium, whereas cells adapted to silicon will not extrude the silicon taken up before, when incubated in a medium containing both elements. References to the possible synthesis of organic silicon compounds resulting from these experiments are discussed.