Molecular cloning and transgenic expression of a synthetic human erythropoietin gene in tobacco

Erythropoietin (EPO) is a hormone belonging to a group of hematopoietic growth factors that control the proliferation and differentiation of bone marrow cells. It induces the production of erythrocytes, thereby increasing the amount of circulating hemoglobin and oxygen. Previous attempts to transgenically express human EPO in plants failed to succeed because the plants exhibited abnormal morphology and infertility. In the present work, we describe the generation of fertile transgenic tobacco plants able to express a synthetic version of human EPO. A 582-bp fragment of the human EPO gene was synthesized using a PCR-based method and ligated into pCR-Blunt. After sequencing, the human EPO fragment was transferred to pWUbi.tm1 and the expression cassette was then transferred to the binary vector pWBVec4a. After Agrobacterium-mediated transformation of Nicotiana tabacum SR1 plants, integration of the transgene into T₀ and T₁ plant genomes was confirmed by PCR. The human EPO gene was found to be expressed in tobacco leaves at the mRNA and protein levels. Self-crossing allowed us to obtain T₁ plants exhibiting Mendelian segregation of the transgene. None of the plants presented any kind of malformation or deformity.     

Thermodynamic, dynamic, structural, and excess entropy anomalies for core-softened potentials

Using molecular dynamic simulations, we study three families of continuous core-softened potentials consisting of two length scales: a shoulder scale and an attractive scale. All the families have the same slope between the two length scales but exhibit different potential energy gap between them. For each family three shoulder depths are analyzed. We show that all these systems exhibit a liquid-liquid phase transition between a high density liquid phase and a low density liquid phase ending at a critical point. The critical temperature is the same for all cases suggesting that the critical temperature is only dependent on the slope between the two scales. The critical pressure decreases with the decrease of the potential energy gap between the two scales suggesting that the pressure is responsible for forming the high density liquid. We also show, using the radial distribution function and the excess entropy analysis, that the density, the diffusion, and the structural anomalies are present if particles move from the attractive scale to the shoulder scale with the increase of the temperature indicating that the anomalous behavior depends only in what happens up to the second coordination shell.     

A lightning initiation mechanism: application to a thunderstorm electrification model

The use of numerical models has greatly increased our understanding of the electrical and microphysical process within electrified clouds. We use the University of Washington, 1.5-dimensional thunderstorm model to examine the effects of including a runaway electron based lightning initiation mechanism. We find that this mechanism can significantly alter the electrification history of modeled storms and produce vertical electric field profiles that are very similar to those of observed storms. To cite this article: R. Solomon et al., C. R. Physique 3 (2002) 1325–1333.     

Paradoxical vocal fold movement: A case report

We present a case of one patient with respiratory stridor that was resolved by speech therapy. Paradoxical vocal fold movements were observed by flexible fiberoptic videolaryngoscopy in this patient during episodes of wheezing and dyspnea. Otherwise, normal vocal fold movement was observed in normal conditions of breathing (out of the crisis) and during phonation. Many different terms have been used to describe this entity in the literature, and it is crucial that clinicians recognize the subtle signs of functional stridor. Paradoxical vocal fold motion has to be considered as an important cause of respiratory disease. Its recognition and treatment are discussed in this report.     

Measurement of vibratioin-vidration pumped population distributions in nitric oxide

A direct-current glow discharge is used to excite flowing mixtures of NO/He and NO/N₂. A strongly vibration-vibration pumped vibrational population distribution over the states NO(X ²Π, υ = 1) to NO(X²Π, υ = 15) is measured by infrared emission spectroscopy. Measured population distributions are consistent with results calculated from earlier measurements of V-V and V-T,R rates.     

Functionalized higher acenes: hexacene and heptacene

We have extended our functionalization strategy for pentacene to the higher acenes hexacene and heptacene. Provided a large enough alkyne substituent is used, these large aromatic rods are both stable and soluble and can be characterized spectroscopically as well as by single-crystal X-ray diffraction.     

Addressing the role of working memory in mathematical word-problem solving when designing intervention for struggling learners

ZDM – Mathematics Education - The focus of this article is the well documented association between low working memory capacity and difficulty with mathematical word-problem solving. We begin...     

Complete assignment of the 1H and 13C NMR spectra of garciniaphenone and keto-enol equilibrium statements for prenylated benzophenones

This article reports the structural elucidation by IR, UV and MS spectroscopic data along with 1H and 13C NMR chemical shift assignments of two benzophenones isolated from the fruit pericarp of Garcinia brasiliensis Mart. (Clusiaceae): garciniaphenone, (1R,5S,7S)-3-benzoyl-4-hydroxy-6,6-dimethyl-5,7-di(3-methyl-2-butenyl)bicyclo[3.3.1]non-3-ene-2,9-dione, a novel triprenylated benzophenone; and 7-epi-clusianone, a tetraprenylated benzophenone that has already been extracted from another species of the same family. Furthermore, the keto-enol tautomeric equilibrium at solution-state was described for these compounds by 1D and 2D NMR spectral methods and one attempt to rationalize the different ratios between the noted tautomers was based on stereochemical features.     

Fluorescence properties and photophysics of the sulfoindocyanine Cy3 linked covalently to DNA

The sulfoindocyanine Cy3 is one of the most commonly used fluorescent dyes in the investigation of the structure and dynamics of nucleic acids by means of fluorescence methods. In this work, we report the fluorescence and photophysical properties of Cy3 attached covalently to single-stranded and duplex DNA. Steady-state and time-resolved fluorescence techniques were used to determine fluorescence quantum yields, emission lifetimes, and fluorescence anisotropy decays. The existence of a transient photoisomer was investigated by means of transient absorption techniques. The fluorescence quantum yield of Cy3 is highest when attached to the 5' terminus of single-stranded DNA (Cy3-5' ssDNA), and decreases by a factor of 2.4 when the complementary strand is annealed to form duplex DNA (Cy3-5' dsDNA). Substantial differences were also observed between the 5'-modified strands and strands modified through an internal amino-modified deoxy uridine. The fluorescence decay of Cy3 became multiexponential upon conjugation to DNA. The longest lifetime was observed for Cy3-5' ssDNA, where about 50% of the decay is dominated by a 2.0-ns lifetime. This value is more than 10 times larger than the fluorescence lifetime of the free dye in solution. These observations are interpreted in terms of a model where the molecule undergoes a trans-cis isomerization reaction from the first excited state. We observed that the activation energy for photoisomerization depends strongly on the microenvironment in which the dye is located. The unusually high activation energy measured for Cy3-5' ssDNA is an indication of dye-ssDNA interactions. In fact, the time-resolved fluorescence anisotropy decay of this sample is dominated by a 2.5-ns rotational correlation time, which evidences the lack of rotational freedom of the dye around the linker that separates it from the terminal 5' phosphate. The remarkable variations in the photophysical properties of Cy3-DNA constructs demonstrate that caution should be used when Cy3 is used in studies employing DNA conjugates.