Mutational analysis of the major soybean UreF paralogue involved in urease activation

The soybean genome duplicated ∼14 and 45 million years ago and has many paralogous genes, including those in urease activation (emplacement of Ni and CO2 in the active site). Activation requires the UreD and UreF proteins, each encoded by two paralogues. UreG, a third essential activation protein, is encoded by the single-copy Eu3, and eu3 mutants lack activity of both urease isozymes. eu2 has the same urease-negative phenotype, consistent with Eu2 being a single-copy gene, possibly encoding a Ni carrier. Unexpectedly, two eu2 alleles co-segregated with missense mutations in the chromosome 2 UreF paralogue (Ch02UreF), suggesting lack of expression/function of Ch14UreF. However, Ch02UreF and Ch14UreF transcripts accumulate at the same level. Further, it had been shown that expression of the Ch14UreF ORF complemented a fungal ureF mutant. A third, nonsense (Q2*) allelic mutant, eu2-c, exhibited 5- to 10-fold more residual urease activity than missense eu2-a or eu2-b, though eu2-c should lack all Ch02UreF protein. It is hypothesized that low-level activation by Ch14UreF is ‘spoiled’ by the altered missense Ch02UreF proteins (‘epistatic dominant-negative’). In agreement with active ‘spoiling’ by eu2-b-encoded Ch02UreF (G31D), eu2-b/eu2-c heterozygotes had less than half the urease activity of eu2-c/eu2-c siblings. Ch02UreF (G31D) could spoil activation by Chr14UreF because of higher affinity for the activation complex, or because Ch02UreF (G31D) is more abundant than Ch14UreF. Here, the latter is favoured, consistent with a reported in-frame AUG in the 5' leader of Chr14UreF transcript. Translational inhibition could represent a form of ‘functional divergence’ of duplicated genes

Experimental observation of optical rotation generated in vacuum by a magnetic field

We report the experimental observation of a light polarization rotation in vacuum in the presence of a transverse magnetic field. Assuming that data distribution is Gaussian, the average measured rotation is (3.9+/-0.5)e-12 rad/pass, at 5 T with 44000 passes through a 1m long magnet, with lambda = 1064 nm. The relevance of this result in terms of the existence of a light, neutral, spin-zero particle is discussed.Comment: 11 pages, 4 figures, submitted to Physical Review Letters Comment to version 2: minor changes to abstract and final discussion. Added 2 references Comment to version 3: corrected minor typographical errors, eliminated the distinction between scalar and pseudoscalar in the particle interpretation of the resul

Dynamical response of the "GGG" rotor to test the Equivalence Principle: theory, simulation and experiment. Part I: the normal modes

Recent theoretical work suggests that violation of the Equivalence Principle might be revealed in a measurement of the fractional differential acceleration $\eta$ between two test bodies -of different composition, falling in the gravitational field of a source mass- if the measurement is made to the level of $\eta\simeq 10^{-13}$ or better. This being within the reach of ground based experiments, gives them a new impetus. However, while slowly rotating torsion balances in ground laboratories are close to reaching this level, only an experiment performed in low orbit around the Earth is likely to provide a much better accuracy. We report on the progress made with the "Galileo Galilei on the Ground" (GGG) experiment, which aims to compete with torsion balances using an instrument design also capable of being converted into a much higher sensitivity space test. In the present and following paper (Part I and Part II), we demonstrate that the dynamical response of the GGG differential accelerometer set into supercritical rotation -in particular its normal modes (Part I) and rejection of common mode effects (Part II)- can be predicted by means of a simple but effective model that embodies all the relevant physics. Analytical solutions are obtained under special limits, which provide the theoretical understanding. A simulation environment is set up, obtaining quantitative agreement with the available experimental data on the frequencies of the normal modes, and on the whirling behavior. This is a needed and reliable tool for controlling and separating perturbative effects from the expected signal, as well as for planning the optimization of the apparatus.Comment: Accepted for publication by "Review of Scientific Instruments" on Jan 16, 2006. 16 2-column pages, 9 figure

EXPERIMENTAL VALIDATION OF A HIGH ACCURACY TEST OF THE EQUIVALENCE PRINCIPLE WITH THE SMALL SATELLITE "GALILEO GALILEI"

The small satellite "Galileo Galilei" (GG) has been designed to test the equivalence principle (EP) to 10-17 with a total mass at launch of 250 kg. The key instrument is a differential accelerometer made up of weakly coupled coaxial, concentric test cylinders rapidly spinning around the symmetry axis and sensitive in the plane perpendicular to it, lying at a small inclination from the orbit plane. The whole spacecraft spins around the same symmetry axis so as to be passively stabilized. The test masses are large (10 kg each, to reduce thermal noise), their coupling is very weak (for high sensitivity to differential effects), and rotation is fast (for high frequency modulation of the signal). A 1 g version of the accelerometer ("Galileo Galilei on the Ground" — GGG) has been built to the full scale — except for coupling, which cannot be as weak as in the absence of weight, and a motor to maintain rotation (not needed in space due to angular momentum conservation). GGG has proved: (i) high Q; (ii) auto-centering and long term stability; (iii) a sensitivity to EP testing which is close to the target sensitivity of the GG experiment provided that the physical properties of the experiment in space are going to be fully exploited

SerpinB3 as hepatic marker of post-resective shear stress

Post-resective liver failure is a frequent complication of liver surgery and it is due to portal hyperperfusion of the remnant liver and to arterial vasoconstriction, as buffer response of the hepatic artery. In this context, splenectomy allows a reduction of portal flow and increases the survival chance in preclinical models. SerpinB3 is over-expressed in the liver in oxidative stress conditions, as a mechanism of cell defense to provide survival by apoptosis inhibition and cell proliferation. In this study, the expression of SerpinB3 was assessed as predictor of liver damage in in vivo models of major hepatic resection with or without splenectomy. Wistar male rats were divided into 4 groups: group A received 30% hepatic resection, group B > 60% resection, group C > 60% resection with splenectomy and group D sham-operated. Before and after surgery liver function tests, echo Doppler ultrasound and gene expression were assessed. Transaminase values and ammonium were significantly higher in groups that underwent major hepatic resection. Echo Doppler ultrasound showed the highest portal flow and resistance of the hepatic artery in the group with > 60% hepatectomy without splenectomy, while the association of splenectomy determined no increase in portal flow and hepatic artery resistance. Only the group of rats without splenectomy showed higher shear-stress conditions, reflected by higher levels of HO-1, Nox1 and of Serpinb3, the latter associated with an increase of IL-6. In conclusion, splenectomy controls inflammation and oxidative damage, preventing the expression of Serpinb3. Therefore, SerpinB3 can be considered as a marker of post-resective shear stress

Optical production and detection of dark matter candidates

The PVLAS collaboration is at present running, at the Laboratori Nazionali di Legnaro of I.N.F.N., Padova, Italy, a very sensitive optical ellipsometer capable of measuring the small rotations or ellipticities which can be acquired by a linearly polarized laser beam propagating in vacuum through a transverse magnetic feld (vacuum magnetic birefringence). The apparatus will also be able to set new limits on mass and coupling constant of light scalar/pseudoscalar particles coupling to two photons by both producing and detecting the hypothetical particles. The axion, introduced to explain parity conservation in strong interactions, is an example of this class of particles, all of which are considered possible dark matter candidates. The PVLAS apparatus consists of a very high finesse (> 140000), 6.4 m long, Fabry-Perot cavity immersed in an intense dipolar magnetic field (~6.5 T). A linearly polarized laser beam is frequency locked to the cavity and analysed, using a heterodyne technique, for rotation and/or ellipticity acquired within the magnetic field.Comment: presented at "Frontier Detectors for Frontier Physics - 8th Pisa Meeting on Advanced Detectors - May 21-27, 2000" to appear in: Nucl.Instr. and Meth.

New PVLAS results and limits on magnetically induced optical rotation and ellipticity in vacuum

IIn 2006 the PVLAS collaboration reported the observation of an optical rotation generated in vacuum by a magnetic field. To further check against possible instrumental artifacts several upgrades to the PVLAS apparatus have been made during the last year. Two data taking runs, at the wavelength of 1064 nm, have been performed in the new configuration with magnetic field strengths of 2.3 T and 5 T. The 2.3 T field value was chosen in order to avoid stray fields. The new observations do not show the presence of a rotation signal down to the levels of $1.2\cdot 10^{-8}$ rad at 5 T and $1.0\cdot 10^{-8}$ rad at 2.3 T (at 95% c.l.) with 45000 passes in the magnetic field zone. In the same conditions no ellipticity signal was detected down to $1.4\cdot 10^{-8}$ at 2.3 T (at 95% c.l.), whereas at 5 T a signal is still present. The physical nature of this ellipticity as due to an effect depending on $B^2$ can be excluded by the measurement at 2.3 T. These new results completely exclude the previously published magnetically induced vacuum dichroism results, indicating that they were instrumental artifacts. These new results therefore also exclude the particle interpretation of the previous PVLAS results as due to a spin zero boson. The background ellipticity at 2.3 T can be used to determine a new limit on the total photon-photon scattering cross section of $\sigma_{\gamma\gamma} < 4.5 \cdot10^{-34}$ barn at 95% c.l..Comment: 25 pages, 7 figures Main changes rel. to v.2: minor changes to abstract, replaced Figures 4,5,6, corrected typographical errors. Paper submitted to Physical Review

Limits on Low Energy Photon-Photon Scattering from an Experiment on Magnetic Vacuum Birefringence

Experimental bounds on induced vacuum magnetic birefringence can be used to improve present photon-photon scattering limits in the electronvolt energy range. Measurements with the PVLAS apparatus (E. Zavattini {\it et al.}, Phys. Rev. D {\bf77} (2008) 032006) at both $\lambda = 1064$ nm and 532 nm lead to bounds on the parameter {\it A$_{e}$}, describing non linear effects in QED, of $A_{e}^{(1064)} < 6.6\cdot10^{-21}$ T$^{-2}$ @ 1064 nm and $A_{e}^{(532)} < 6.3\cdot10^{-21}$ T$^{-2}$ @ 532 nm, respectively, at 95% confidence level, compared to the predicted value of $A_{e}=1.32\cdot10^{-24}$ T$^{-2}$. The total photon-photon scattering cross section may also be expressed in terms of $A_e$, setting bounds for unpolarized light of $\sigma_{\gamma\gamma}^{(1064)} < 4.6\cdot10^{-62}$ m$^{2}$ and $\sigma_{\gamma\gamma}^{(532)} < 2.7\cdot10^{-60}$ m$^{2}$. Compared to the expected QED scattering cross section these results are a factor of $\simeq2\cdot10^{7}$ higher and represent an improvement of a factor about 500 on previous bounds based on ellipticity measurements and of a factor of about $10^{10}$ on bounds based on direct stimulated scattering measurements

Signal Processing in the PVLAS Experiment

Nonlinear interactions of light with light are well known in quantum electronics, and it is quite common to generate harmonic or subharmonic beams from a primary laser with photonic crystals. One suprising result of quantum electrodynamics is that because of the quantum fluctuations of charged fields, the same can happen in vacuum. The virtual charged particle pairs can be polarized by an external field and vacuum can thus become birefringent: the PVLAS experiment was originally meant to explore this strange quantum regime with optical methods. Since its inception PVLAS has found a new, additional goal: in fact vacuum can become a dichroic medium if we assume that it is filled with light neutral particles that couple to two photons, and thus PVLAS can search for exotic particles as well. PVLAS implements a complex signal processing scheme: here we describe the double data acquisition chain and the data analysis methods used to process the experimental data.Comment: presented by E. Milotti to the WSEAS-ISCGAV '05 Conference, Malta, 15-17/9/200