The phosphoinositide PI(3,5)P2 mediates activation of mammalian but not plant TPC proteins: functional expression of endolysosomal channels in yeast and plant cells

Two-pore channel proteins (TPC) encode intracellular ion channels in both animals and plants. In mammalian cells, the two isoforms (TPC1 and TPC2) localize to the endo-lysosomal compartment, whereas the plant TPC1 protein is targeted to the membrane surrounding the large lytic vacuole. Although it is well established that plant TPC1 channels activate in a voltage- and calcium-dependent manner in vitro, there is still debate on their activation under physiological conditions. Likewise, the mode of animal TPC activation is heavily disputed between two camps favoring as activator either nicotinic acid adenine dinucleotide phosphate (NAADP) or the phosphoinositide PI(3,5)P₂. Here, we investigated TPC current responses to either of these second messengers by whole-vacuole patch-clamp experiments on isolated vacuoles of Arabidopsis thaliana. After expression in mesophyll protoplasts from Arabidopsis tpc1 knock-out plants, we detected the Arabidopsis TPC1-EGFP and human TPC2-EGFP fusion proteins at the membrane of the large central vacuole. Bath (cytosolic) application of either NAADP or PI(3,5)P₂ did not affect the voltage- and calcium-dependent characteristics of AtTPC1-EGFP. By contrast, PI(3,5)P₂ elicited large sodium currents in hTPC2-EGFP-containing vacuoles, while NAADP had no such effect. Analogous results were obtained when PI(3,5)P₂ was applied to hTPC2 expressed in baker’s yeast giant vacuoles. Our results underscore the fundamental differences in the mode of current activation and ion selectivity between animal and plant TPC proteins and corroborate the PI(3,5)P₂-mediated activation and Na⁺ selectivity of mammalian TPC2.     

Multigenic control of Listeria monocytogenes susceptibility in mice

We have used a novel quantitative trait locus model to study the genetics of survival of F2 progeny of susceptible BALB/cByJ and resistant C57BL/6ByJ mice that have been infected with Listeria monocytogenes. This allowed us to map modifiers of L. monocytogenes susceptibility to chromosomes 5 and 13.     

System identification and validation for output prediction of a dynamic hydrologic process

The paper presents an identification procedure for a dynamic model of am hydrologic process. The process involves solute transport in streams subject to aquifer interaction and unsteady flows and the intended use of the model is prediction. Detailed assumptions and results are provided to illustrate the level of comprehensive analysis required to assess model adequacy. The assessment procedure easily generalizes to any dynamic model which is linear-in-the-parameters. As a fundamental tool, instrumental variable algorithms can be adopted which have a number of attractive features. These algorithms make both model-order identification and specification among alternatives a straightforward task. They are known to be consistent estimators in the presence of a wide class of errors. It is seen that they can be made stable and robust in the presence of data outliers. Instrumental variable algorithms can also be used which are asymptotically efficient and provide a covariance matrix of parameter estimates. The paper shows how they aid the quantification of predictive uncertainty and investigates the validity of the underlying assumptions. Further, it illustrates that, when instrumental variable algorithms are used in recursive mode, they can be used not only as an additional tool to access model inadequacy but also as an aid to model improvements.     

Production of antibodies against synthetic angiotensin II in various animal species

Zusammenfassung Bei Ratten und Kaninchen, jedoch nicht bei Hunden und Katzen, konnten durch wiederholte Injektion von Angiotensin-II-Amid-Carbodiimid-Proteinkomplex in Freund'schem Adjuvans zirkulierende Antikörper gegen Angiotensin erzeugt werden. Durch diese Immunisierung wurde die pressorische Wirkung von exogenem Angiotensin aufgehoben bzw. abgeschwächt, während die von Noradrenalin oder Vasopressin unbeeinflusst blieb. Der Reningehalt der Nieren von Ratten und Kaninchen wurde durch die Immunisierung nicht verändert.     

A filament of dark matter between two clusters of galaxies

It is a firm prediction of the concordance cold-dark-matter cosmological model that galaxy clusters occur at the intersection of large-scale structure filaments. The thread-like structure of this 'cosmic web' has been traced by galaxy redshift surveys for decades. More recently, the warm–hot intergalactic medium (a sparse plasma with temperatures of 10(5) kelvin to 10(7) kelvin) residing in low-redshift filaments has been observed in emission and absorption. However, a reliable direct detection of the underlying dark-matter skeleton, which should contain more than half of all matter, has remained elusive, because earlier candidates for such detections were either falsified or suffered from low signal-to-noise ratios and unphysical misalignments of dark and luminous matter. Here we report the detection of a dark-matter filament connecting the two main components of the Abell 222/223 supercluster system from its weak gravitational lensing signal, both in a non-parametric mass reconstruction and in parametric model fits. This filament is coincident with an overdensity of galaxies and diffuse, soft-X-ray emission, and contributes a mass comparable to that of an additional galaxy cluster to the total mass of the supercluster. By combining this result with X-ray observations, we can place an upper limit of 0.09 on the hot gas fraction (the mass of X-ray-emitting gas divided by the total mass) in the filament.     

Investigation on regenerator temperature inhomogeneity in Stirling-type pulse tube cooler

Regenerator is one of the most crucial components to pulse tube cooler (PTC) and thermoacoustic engine. As such regenerator is scaled up to high-power, the thermal and hydrodynamic communication transverse to the acoustic axis gets weaker and weaker. Under this condition, any unsymmetric factor could cause serious instability to the cooler or engine, which degrades their performance. Investigation has been carried out on a high-power two-stage thermal-coupled U-shape Stirling-type PTC. By detailed circumferential temperature measurements along the middle heat exchanger and second stage regen-erator, a kind of temperature inhomogeneity caused by unsymmetric pre-cooling effect of inter-stage thermal bridge was found in the lower part of the regenerator of the PTC. The temperature inhomoge-neity originating from the middle heat exchanger of the second stage regenerator amplified itself in the lower part of the regenerator and then internal streaming formed. The maximal radial temperature dif-ference could reach 30―40 K. Experimental results show that the temperature inhomogeneity intensi-fies with increased pre-cooling power and its direction can be reversed by changing the pre-cooling effect of the first stage PTC to heating effect by using external thermal load. This research shows that it is important to maintain the heating or cooling effects of heat exchangers uniform in high-power re-generative coolers and engines.