GTP and Ca2+ Modulate the Inositol 1,4,5-Trisphosphate-Dependent Ca2+ Release in Streptolysin O-Permeabilized Bovine Adrenal Chromaffin Cells

The inositol 1,4,5-trisphosphate (IP3)-induced Ca2+ release was studied using streptolysin O-permeabilized bovine adrenal chromaffin cells. The IP3-induced Ca2+ release was followed by Ca2+ reuptake into intracellular compartments. The IP3-induced Ca2+ release diminished after sequential applications of the same amount of IP3. Addition of 20 μM GTP fully restored the sensitivity to IP3. Guanosine 5'-O-(3-thio)triphosphate (GTPγS) could not replace GTP but prevented the action of GTP. The effects of GTP and GTPγS were reversible. Neither GTP nor GTPγS induced release of Ca2+ in the absence of IP3. The amount of Ca2+ whose release was induced by IP3 depended on the free Ca2+ concentration of the medium. At 0.3 μM free Ca2+, a half-maximal Ca2+ release was elicited with ∼0.1 μM IP3. At 1 μM free Ca2+, no Ca2+ release was observed with 0.1 μM IP3; at this Ca2+ concentration, higher concentrations of IP3 (0.25 μM) were required to evoke Ca2+ release. At 8 μM free Ca2+, even 0.25 μM IP3 failed to induce release of Ca2+ from the store. The IP3-induced Ca2+ release at constant low (0.2 μM) free Ca2+ concentrations correlated directly with the amount of stored Ca2+. Depending on the filling state of the intracellular compartment, 1 mol of IP3 induced release of between 5 and 30 mol of Ca2+

SOAP-based services provided by the European Bioinformatics Institute

SOAP (Simple Object Access Protocol) () based Web Services technology () has gained much attention as an open standard enabling interoperability among applications across heterogeneous architectures and different networks. The European Bioinformatics Institute (EBI) is using this technology to provide robust data retrieval and data analysis mechanisms to the scientific community and to enhance utilization of the biological resources it already provides [N. Harte, V. Silventoinen, E. Quevillon, S. Robinson, K. Kallio, X. Fustero, P. Patel, P. Jokinen and R. Lopez (2004) Nucleic Acids Res., 32, 3–9]. These services are available free to all users from

Functional improvement and maturation of rat and human engineered heart tissue by chronic electrical stimulation

Spontaneously beating engineered heart tissue (EHT) represents an advanced in vitro model for drug testing and disease modeling, but cardiomyocytes in EHTs are less mature and generate lower forces than in the adult heart. We devised a novel pacing system integrated in a setup for videooptical recording of EHT contractile function over time and investigated whether sustained electrical field stimulation improved EHT properties. EHTs were generated from neonatal rat heart cells (rEHT, n=96) or human induced pluripotent stem cell (hiPSC)-derived cardiomyocytes (hEHT, n=19). Pacing with biphasic pulses was initiated on day 4 of culture. REHT continuously paced for 16-18 days at 0.5Hz developed 2.2× higher forces than nonstimulated rEHT. This was reflected by higher cardiomyocyte density in the center of EHTs, increased connexin-43 abundance as investigated by two-photon microscopy and remarkably improved sarcomere ultrastructure including regular M-bands. Further signs of tissue maturation include a rightward shift (to more physiological values) of the Ca(2+)-response curve, increased force response to isoprenaline and decreased spontaneous beating activity. Human EHTs stimulated at 2Hz in the first week and 1.5Hz thereafter developed 1.5× higher forces than nonstimulated hEHT on day 14, an ameliorated muscular network of longitudinally oriented cardiomyocytes and a higher cytoplasm-to-nucleus ratio. Taken together, continuous pacing improved structural and functional properties of rEHTs and hEHTs to an unprecedented level. Electrical stimulation appears to be an important step toward the generation of fully mature EHT

The sizes of mini-voids in the local universe: an argument in favor of a warm dark matter model?

Using high-resolution simulations within the Cold and Warm Dark Matter models we study the evolution of small scale structure in the Local Volume, a sphere of 8 Mpc radius around the Local Group. We compare the observed spectrum of mini-voids in the Local Volume with the spectrum of mini-voids determined from the simulations. We show that the \LWDM model can easily explain both the observed spectrum of mini-voids and the presence of low-mass galaxies observed in the Local Volume, provided that all haloes with circular velocities greater than 20 km/s host galaxies. On the contrary within the LCDM model the distribution of the simulated mini-voids reflects the observed one if haloes with maximal circular velocities larger than 35 km/s host galaxies. This assumption is in contradiction with observations of galaxies with circular velocities as low as 20 km/s in our Local Universe. A potential problem of the LWDM model could be the late formation of the haloes in which the gas can be efficiently photo-evaporated. Thus star formation is suppressed and low-mass haloes might not host any galaxy at all.Comment: 13 pages, 10 figures, version 2, subsection 3.1 added, accepted to MNRA

Narrow genetic base in forest restoration with holm oak (Quercus ilex L.) in Sicily

In order to empirically assess the effect of actual seed sampling strategy on genetic diversity of holm oak (Quercus ilex) forestations in Sicily, we have analysed the genetic composition of two seedling lots (nursery stock and plantation) and their known natural seed origin stand by means of six nuclear microsatellite loci. Significant reduction in genetic diversity and significant difference in genetic composition of the seedling lots compared to the seed origin stand were detected. The female and the total effective number of parents were quantified by means of maternity assignment of seedlings and temporal changes in allele frequencies. Extremely low effective maternity numbers were estimated (Nfe $\approx$ 2-4) and estimates accounting for both seed and pollen donors gave also low values (Ne $\approx$ 35-50). These values can be explained by an inappropriate forestry seed harvest strategy limited to a small number of spatially close trees

A Keck/DEIMOS spectroscopic survey of faint Galactic satellites: searching for the least massive dwarf galaxies

[abridged] We present the results of a spectroscopic survey of the recently discovered faint Milky Way satellites Boo, UMaI, UMaII and Wil1. Using the DEIMOS spectrograph on Keck, we have obtained samples that contain from 15 to 85 probable members of these satellites for which we derive radial velocities precise to a few km/s down to i~21-22. About half of these stars are observed with a high enough S/N to estimate their metallicity to within \pm0.2 dex. From this dataset, we show that UMaII is the only object that does not show a clear radial velocity peak. However, the measured systemic radial velocity (v_r=115\pm5 km/s) is in good agreement with recent simulations in which this object is the progenitor of the recently discovered Orphan Stream. The three other satellites show velocity dispersions that make them highly dark-matter dominated systems. In particular the Willman 1 object is not a globular cluster given its metallicity scatter over -2.0<[Fe/H]<-1.0 and is therefore almost certainly a dwarf galaxy or dwarf galaxy remnant. We measure a radial velocity dispersion of only 4.3_{-1.3}^{+2.3} km/s around a systemic velocity of -12.3\pm2.3 km/s which implies a mass-to-light ratio of ~700 and a total mass of ~5x10^5 Msun for this satellite, making it the least massive satellite galaxy known to date. Such a low mass could mean that the 10^7 Msun limit that had until now never been crossed for Milky Way and Andromeda satellite galaxies may only be an observational limit and that fainter, less massive systems exist within the Local Group. However, more modeling and an extended search for potential extra-tidal stars are required to rule out the possibility that these systems have not been significantly heated by tidal interaction.Comment: 24 pages, 11 figures, MNRAS accepte

HIV-1 DNA predicts disease progression and post-treatment virological control.

In HIV-1 infection, a population of latently infected cells facilitates viral persistence despite antiretroviral therapy (ART). With the aim of identifying individuals in whom ART might induce a period of viraemic control on stopping therapy, we hypothesised that quantification of the pool of latently infected cells in primary HIV-1 infection (PHI) would predict clinical progression and viral replication following ART. We measured HIV-1 DNA in a highly characterised randomised population of individuals with PHI. We explored associations between HIV-1 DNA and immunological and virological markers of clinical progression, including viral rebound in those interrupting therapy. In multivariable analyses, HIV-1 DNA was more predictive of disease progression than plasma viral load and, at treatment interruption, predicted time to plasma virus rebound. HIV-1 DNA may help identify individuals who could safely interrupt ART in future HIV-1 eradication trials