Efficiency of transient transformation in tobacco protoplasts is independent of plasmid amount

An optimized protocol for the transient transformation of tobacco protoplasts mediated by polyethylene\u2013glycol (PEG) is here described. As expected, the quantitative b\u2013glucuronidase (Gus) activity driven by pCaMVGus was dependent on the amount of plasmid used. Nevertheless, we demonstrate by an immunodetection method that transformation efficiency did not depend on the amount of plasmid used but on the limitation imposed by cell competence. In fact, we obtained the same percentage of transformed cells (about 60%) using a wide range of plasmid concentrations (0.1\u201310 mg per test). Finally, we show that, when we used two plasmid types in a mixture at a concentration ranging from 0.1 to 10 mg for each, all transformed cells expressed proteins encoded by both plasmids. Transient expression and co-transformation experiments are routinely used methods and, probably, the major results from this work were assumed by many researchers in this field, but our data experimentally support this assumption

The product of the rice myb7 unspliced mRNA dimerizes with the maize leucine-zipper opaque2 and stimulates its activity in transient expression assay

myb7 mRNA is present in rice in spliced and unspliced forms, splicing being enhanced by anoxia. The protein (Mybleu) encoded by the unspliced mRNA is composed of an incomplete Myb domain followed by a leucine zipper; however, it lacks canonical sequences for DNA binding, transcriptional activation, and nuclear localization. We show here that in transiently transformed tobacco protoplasts, Mybleu is able to enhance the transcriptional activity of the maize leucine zipper Opaque2 on its target b32 promoter. The Mybleu transactivation effect is strictly dependent on the presence of Opaque2 and is driven by Mybleu-Opaque2 heterodimers. Mybleu is located in the nucleus, both in rice and in transformed tobacco protoplasts. In rice, the protein is expressed in regions corresponding to undifferentiated cells of roots and coleoptiles. Therefore, myb7 mRNA encodes, depending on its splicing, two transcription factors belonging to separate classes. One of them, Myblue, has novel structural characteristics, suggesting the existence of new mechanisms acting in the activation of transcription. -------------------------------------------------------------------------------

Overexpression of Osmyb4 enhances compatible solute accumulation and increases stress tolerance of arabidopsis thaliana

In this paper, we report the metabolic and molecular changes in response to cold and drought induced in Osmyb4 transgenic Arabidopsis thaliana compared with the wildtype (WT). The rice Osmyb4 gene codes for a transcription factor (Myb4) induced by cold treatment and, in Arabidopsis transgenic plants, improves cold and freezing tolerance [Vannini C, Locatelli F, Bracale M, Magnani E, Marsoni M, Osnato M, Mattana M, Baldoni E, Coraggio I (2004) Plant J 37: 115-127]. Here, we report the ability of Myb4 to induce also drought tolerance in Arabidopsis transgenic plants. By the use of nuclear magnetic resonance (NMR) and enzymatic assays, we showed that several compatible solutes (glucose, fructose, sucrose, proline, glycine betaine and sinapoyl malate) accumulate in higher amount in Osmyb4-overexpressing plants with respect to the WT, both under normal and stress conditions. Considering proline, we also found that in transgenic plants the levels of the mRNAs coding for \u3941- pyrroline-5-carboxylate synthase (EC not assigned) and for \u3941- pyrroline-5-carboxylate dehydrogenase (EC 1.5.1.12) were higher and lower, respectively. The constitutive activation of several stress-inducible pathways and different kinetics in the accumulation of several metabolites, in Myb4 transgenic plants, may represent an advantage to prepare plants to face the stress condition. Moreover, these results taken together suggest that Myb4 integrates the activation of multiple components of stress response

Nuclear Structure Calculations and Modern Nucleon-Nucleon Potentials

We study ground-state properties of the doubly magic nuclei 4He, 16O, and 40Ca employing the Goldstone expansion and using as input four different high-quality nucleon-nucleon (NN) potentials. The short-range repulsion of these potentials is renormalized by constructing a smooth low-momentum potential V-low-k. This is used directly in a Hartree-Fock approach and corrections up to third order in the Goldstone expansion are evaluated. Comparison of the results shows that they are only slightly dependent on the choice of the NN potential.Comment: 5 pages, submitted to Physical Review

Computational Nuclear Physics and Post Hartree-Fock Methods

We present a computational approach to infinite nuclear matter employing Hartree-Fock theory, many-body perturbation theory and coupled cluster theory. These lectures are closely linked with those of chapters 9, 10 and 11 and serve as input for the correlation functions employed in Monte Carlo calculations in chapter 9, the in-medium similarity renormalization group theory of dense fermionic systems of chapter 10 and the Green's function approach in chapter 11. We provide extensive code examples and benchmark calculations, allowing thereby an eventual reader to start writing her/his own codes. We start with an object-oriented serial code and end with discussions on strategies for porting the code to present and planned high-performance computing facilities.Comment: 82 pages, to appear in Lecture Notes in Physics (Springer), "An advanced course in computational nuclear physics: Bridging the scales from quarks to neutron stars", M. Hjorth-Jensen, M. P. Lombardo, U. van Kolck, Editor

Effective interactions and operators in no-core shell model

Solutions to the nuclear many-body problem rely on effective interactions, and in general effective operators, to take into account effects not included in calculations. These include effects due to the truncation to finite model spaces where a numerical calculation is tractable, as well as physical terms not included in the description in the first place. In the no-core shell model (NCSM) framework, we discuss two approaches to the effective interactions based on (i) unitary transformations and (ii) effective field theory (EFT) principles. Starting from a given Hamiltonian, the unitary transformation approach is designed to take into account effects induced by the truncation to finite model spaces in which a numerical calculation is performed. This approach was widely applied to the description of nuclear properties of light nuclei; we review the theory and present representative results. In the EFT approach, a Hamiltonian is always constructed in a truncated model space according to the symmetries of the underlying theory, making use of power counting to limit the number of interactions included in the calculations. Hence, physical terms not explicitly included in the calculation are treated on the same footing with the truncation to a finite model space. In this approach, we review results for both nuclear and trapped atomic systems, for which the effective theories are formally similar, albeit describing different underlying physics. Finally, the application of the EFT method of constructing effective interactions to Gamow shell model is briefly discussed.Comment: 56 pages, 26 figure

Clinical features of patients with type 2 diabetes with and without Covid-19: a case control study (CoViDiab I)

Aims: To evaluate whether subjects with diabetes hospitalized for Coronavirus disease-19 (Covid-19) represent a subgroup of patients with high-risk clinical features compared to patients with diabetes without Covid-19. Methods: In this case-control study 79 patients with type 2 diabetes out of 354 adults hospitalized for Covid-19 and 158 controls with type 2 diabetes but without Covid-19, matched for age and gender, were enrolled. Medical history and concomitant therapies were retrieved from medical charts and compared between cases and controls, controlling for confounders. Results: Fully-adjusted multivariate logistic regression model showed that previous CVD history did not differ between patients with and without Covid-19 (odds ratio 1.40, 95% confidence interval [CI]: 0.59–3.32, p = 0.45). A higher prevalence of chronic obstructive pulmonary disease (COPD) (OR 3.72, 95%CI: 1.42–9.72, p = 0.007) and of chronic kidney disease (CKD) (OR 3.08, 95%CI: 1.18–8.06, p = 0.022) and a lower prevalence of ever smokers (OR 0.30, 95%CI: 0.13–0.67, p = 0.003), of users of lipid lowering agents (OR 0.26, 95%CI: 0.12–0.54, p < 0.001), and of anti-hypertensive drugs (OR 0.39, 95%CI: 0.16–0.93, p = 0.033) were found among cases. Conclusions: CVD prevalence does not differ between people with diabetes with and without Covid-19 requiring hospitalization. An increased prevalence of COPD and of CKD in Covid-19 patients with type 2 diabetes is suggested. These findings aid to clarify the relationship between underlying conditions and SARS-CoV-2 infection in the high-risk group of patients with diabetes

Gamow Shell Model Description of Weakly Bound Nuclei and Unbound Nuclear States

We present the study of weakly bound, neutron-rich nuclei using the nuclear shell model employing the complex Berggren ensemble representing the bound single-particle states, unbound Gamow states, and the non-resonant continuum. In the proposed Gamow Shell Model, the Hamiltonian consists of a one-body finite depth (Woods-Saxon) potential and a residual two-body interaction. We discuss the basic ingredients of the Gamow Shell Model. The formalism is illustrated by calculations involving {\it several} valence neutrons outside the double-magic core: $^{6-10}$He and $^{18-22}$O.Comment: 19 pages, 20 encapsulated PostScript figure

Many-body perturbation calculation of spherical nuclei with a separable monopole interaction: I. Finite nuclei

We present calculations of ground state properties of spherical, doubly closed-shell nuclei from $^{16}$O to $^{208}$Pb employing the techniques of many-body perturbation theory using a separable density dependent monopole interaction. The model gives results in Hartree-Fock order which are of similar quality to other effective density-dependent interactions. In addition, second and third order perturbation corrections to the binding energy are calculated and are found to contribute small, but non-negligible corrections beyond the mean-field result. The perturbation series converges quickly, suggesting that this method may be used to calculate fully correlated wavefunctions with only second or third order perturbation theory. We discuss the quality of the results and suggest possible methods of improvement.Comment: 20 Pages, 11 figure