Genetic control of rhizosheath formation in pearl millet.

The rhizosheath, the layer of soil that adheres strongly to roots, influences water and nutrients acquisition. Pearl millet is a cereal crop that plays a major role for food security in arid regions of sub-Saharan Africa and India. We previously showed that root-adhering soil mass is a heritable trait in pearl millet and that it correlates with changes in rhizosphere microbiota structure and functions. Here, we studied the correlation between root-adhering soil mass and root hair development, root architecture, and symbiosis with arbuscular mycorrhizal fungi and we analysed the genetic control of this trait using genome wide association (GWAS) combined with bulk segregant analysis and gene expression studies. Root-adhering soil mass was weakly correlated only to root hairs traits in pearl millet. Twelve QTLs for rhizosheath formation were identified by GWAS. Bulk segregant analysis on a biparental population validated five of these QTLs. Combining genetics with a comparison of global gene expression in the root tip of contrasted inbred lines revealed candidate genes that might control rhizosheath formation in pearl millet. Our study indicates that rhizosheath formation is under complex genetic control in pearl millet and suggests that it is mainly regulated by root exudation.

Value of PET/CT 3D visualization of head and neck squamous cell carcinoma extended to mandible.

PURPOSE: To study an original 3D visualization of head and neck squamous cell carcinoma extending to the mandible by using [18F]-NaF PET/CT and [18F]-FDG PET/CT imaging along with a new innovative FDG and NaF image analysis using dedicated software. The main interest of the 3D evaluation is to have a better visualization of bone extension in such cancers and that could also avoid unsatisfying surgical treatment later on. PATIENTS AND METHODS: A prospective study was carried out from November 2016 to September 2017. Twenty patients with head and neck squamous cell carcinoma extending to the mandible (stage 4 in the UICC classification) underwent [18F]-NaF and [18F]-FDG PET/CT. We compared the delineation of 3D quantification obtained with [18F]-NaF and [18F]-FDG PET/CT. In order to carry out this comparison, a method of visualisation and quantification of PET images was developed. This new approach was based on a process of quantification of radioactive activity within the mandibular bone that objectively defined the significant limits of this activity on PET images and on a 3D visualization. Furthermore, the spatial limits obtained by analysis of the PET/CT 3D images were compared to those obtained by histopathological examination of mandibular resection which confirmed intraosseous extension to the mandible. RESULTS: The [18F]-NaF PET/CT imaging confirmed the mandibular extension in 85% of cases and was not shown in [18F]-FDG PET/CT imaging. The [18F]-NaF PET/CT was significantly more accurate than [18F]-FDG PET/CT in 3D assessment of intraosseous extension of head and neck squamous cell carcinoma. This new 3D information shows the importance in the imaging approach of cancers. All cases of mandibular extension suspected on [18F]-NaF PET/CT imaging were confirmed based on histopathological results as a reference. CONCLUSIONS: The [18F]-NaF PET/CT 3D visualization should be included in the pre-treatment workups of head and neck cancers. With the use of a dedicated software which enables objective delineation of radioactive activity within the bone, it gives a very encouraging results. The [18F]-FDG PET/CT appears insufficient to confirm mandibular extension. This new 3D simulation management is expected to avoid under treatment of patients with intraosseous mandibular extension of head and neck cancers. However, there is also a need for a further study that will compare the interest of PET/CT and PET/MRI in this indication.

Differences in meristem size and expression of branching genes are associated with variation in panicle phenotype in wild and domesticated African rice.

BACKGROUND: The African rice Oryza glaberrima was domesticated from its wild relative Oryza barthii about 3000 years ago. During the domestication process, panicle complexity changed from a panicle with low complexity in O. barthii, to a highly branched panicle carrying more seeds in O. glaberrima. To understand the basis of this differential panicle development between the two species, we conducted morphological and molecular analyses of early panicle development. RESULTS: Using X-ray tomography, we analyzed the morphological basis of early developmental stages of panicle development. We uncovered evidence for a wider rachis meristem in O. glaberrima than in O. barthii. At the molecular level, spatial and temporal expression profiles of orthologs of O. sativa genes related to meristem activity and meristem fate control were obtained using in situ hybridization and qRT-PCR. Despite highly conserved spatial expression patterns between O. glaberrima and O. barthii, differences in the expression levels of these early acting genes were detected. CONCLUSION: The higher complexity of the O. glaberrima panicle compared to that of its wild relative O. barthii is associated with a wider rachis meristem and a modification of expression of branching-related genes. Our study indicates that the expression of genes in the miR156/miR529/SPL and TAW1 pathways, along with that of their target genes, is altered from the unbranched stage of development. This suggests that differences in panicle complexity between the two African rice species result from early alterations to gene expression during reproductive development.

miR2118-triggered phased siRNAs are differentially expressed during the panicle development of wild and domesticated African rice species.

BACKGROUND: Rice exhibits a wide range of panicle structures. To explain these variations, much emphasis has been placed on changes in transcriptional regulation, but no large-scale study has yet reported on changes in small RNA regulation in the various rice species. To evaluate this aspect, we performed deep sequencing and expression profiling of small RNAs from two closely related species with contrasting panicle development: the cultivated African rice Oryza glaberrima and its wild relative Oryza barthii. RESULTS: Our RNA-seq analysis revealed a dramatic difference between the two species in the 21 nucleotide small RNA population, corresponding mainly to miR2118-triggered phased siRNAs. A detailed expression profiling during the panicle development of O. glaberrima and O. barthii using qRT-PCRs and in situ hybridization, confirmed a delayed expression of the phased siRNAs as well as their lncRNA precursors and regulators (miR2118 and MEL1 gene) in O. glaberrima compared to O. barthii. We provide evidence that the 21-nt phasiRNA pathway in rice is associated with male-gametogenesis but is initiated in spikelet meristems. CONCLUSION: Differential expression of the miR2118-triggered 21-nt phasiRNA pathway between the two African rice species reflects differential rates of determinate fate acquisition of panicle meristems between the two species.

Normalization of stress and delayed thallium-201 myocardial SPECT: where is the normal reference area?

We describe an unusual uptake pattern in a thallium SPECT study performed after dipyridamole infusion in a patient with a documented history of prior inferior infarction and recent typical chest pain. The stress study exhibited maximum uptake in the inferior wall. The delayed study showed an inferior defect more consistent with the notion of inferior necrosis, with a maximum uptake in the anterior wall. The authors propose a pathophysiologic interpretation consistent with coronary angiography findings, based on the assumption of coronary steal suggested by the occurrence of chest pain at the end of the dipyridamole infusion. The problem of selecting myocardial normal reference area(s) necessary to normalization prior to quantitative comparison stress and delayed studies is discussed.

Synthesis, characterization and biodistribution of new 99mTc Oxo and nitrido complexes of unsaturated tetradentate (N2S2)ligands.

Three unsaturated Schiff base tetradentate (N2S2 or N2SO) ligands were synthesized and characterized. Oxo and nitrido 99m-technetium complexes were obtained with these ligands. The nitrido complexes were formed using a new easy method available as a kit. When injected into rats and mice, these lipophilic complexes were able to cross the blood-brain barrier but brain perfusion imaging could not be performed due to the insufficient uptake and retention time.

[A simulation method for studying scintillation camera collimators]. / Une méthode d'étude par simulation des collimateurs des caméras à scintillation.

This paper presents a computer simulation of photon interaction with collimator septa, which allows the point spread function of scintillation camera collimators to be calculated. The method simulates photon attenuation along their propagation direction in a determinist way. Using this simulation, the spatial resolution, geometric efficiency and penetration index of collimators may be easily assessed. Results obtained with this method are presented and compared with those obtained from standard formulae. We show the usefulness of the simulation which precisely accounts for effects of septum penetration. Measurements performed on two collimators with 131I and 99Tcm point sources provide results consistent with those obtained from the simulation method. In conclusion we show that this method is an accurate tool to assist conception of collimators for nuclear medicine.

[Simulation study of the influence of collimator defects on the uniformity of scintigraphic images]. / Etude en simulation de l'influence des défauts des collimateurs sur l'uniformité des images scintigraphiques.

The mechanical tolerances in building collimators for scintillation cameras are studied. A simulation method has been used to quantify the effects of defects in hole inclination and hole diameter on the uniformity of planar and tomographic images. The calculation takes into account the geometry of the hexagonal hole collimator, the camera intrinsic resolution, the object size, the pixel size, the effect of low-pass filtering, as well as the type, size and position of the defect. For instance, a 0.03 mm diameter defect on several holes located in the central region of a very high resolution collimator can result in a 12% uniformity artefact in tomographic imaging of an 18 cm diameter cylinder, using a 3.45 mm resolution camera, 4.5 mm pixel size, and Hamming filtering with a Nyquist frequency cut-off. A 0.17 degree inclination defect of a few holes can result in the same uniformity artefact. These results show that the building of a collimator has to be very precise.

[Characterization of the response of hexagonal parallel-hole collimators of scintillation cameras]. / Caractérisation de la réponse des collimateurs à trous parallèles hexagonaux des caméras à scintillation.

This paper presents a formulation of the frequency response of hexagonal parallel-hole collimator scintillation cameras. To describe this response, we propose an equation determined semi-empirically from a great number of simulations. The utility of the equation is that it enables the simple calculation of the response from collimator characteristics by taking into account the collimator's hexagonal structure. Because the equation does not assume translation invariance, the results can be directly compared with experimental measurements obtained with a point source. It is particularly interesting for collimators with large holes, like the medium-resolution ones used for high-energy radiation. Quality control and physical performance measurements are thus facilitated for this kind of collimator. Also, we present a new parameter that gives a quantitative assessment of the importance of partition penetration. This parameter is measured directly from the collimator frequency response. It has been studied by simulation, taking into account gamma photon attenuation in collimator partitions. The experimental measurements that have been made are in accord with the proposed equations.

Light and developmental regulation of the Anp-controlled anthocyanin phenotype of bean pods.

In the presence of the dominant allele of the Anp gene, bean pods present a purple-mottled phenotype. The purple pigmentation is variable from cell to cell in the pod epidermal layer and develops as a random mosaic. Three anthocyanidins, delphinidin, petunidin and malvidin, are involved in this purple pigmentation. Anthocyanins accumulated in vacuoles; anthocyanoplasts and cristal bodies were also observed occasionally. A developmental switch is a prerequisite for anthocyanin accumulation in the pods. This does not occur before day 4 after pollination and is controlled by light in competent pods. mRNAs for PAL, CHS, CHI, DFR and UFGT are induced in the pods, indicating that the general anthocyanin biosynthetic pathway is well conserved at both the biochemical and molecular levels in this species. mRNA steady-state level studies of PAL and CHS suggest that the light regulation occurs at the transcriptional level.

Plant bZIP G-box binding factors. Modular structure and activation mechanisms.

In this review we sum-up the knowledge about bZIP G-box binding factors (GBFs), which possess an N-terminal, proline-rich domain. The GBF has been one of the most extensively studied transcription factor family. Based on protein sequence homology with yeast and animal basic leucine-zipper (bZIP) transcription factors, bioinformatic studies have identified their main structural domains (proline-rich, basic and leucine-zipper), which have been further functionally characterized by in vitro and in vivo experiments. Recent reports have led to the discovery of other GBF-specific short amino-acid sequences that may take part in the regulation of gene expression by post-transcriptional modifications or interaction with other proteins such as bZIP enhancing factors or plant 14-3-3-like proteins. We identified a GBF region, called the 'multifunctional mosaic region', that may be implicated in cytoplasmic retention, translocation to the nucleus and regulation of transcription. We also identified many conserved protein motifs that suggest a modular structure for GBFs. At the whole plant level, GBFs have been shown to be involved in developmental and physiological processes in response to major cues such as light or hormones. Nevertheless, it remains difficult to assign a physiological role to a particular GBF protein modular structure. Finally, bringing together these different aspects of GBF studies we propose a model describing the puzzling transduction pathway involving GBFs from cytoplasmic events of signal transduction to the regulation of gene expression in the nucleus.

Flow Cytometric Fluorescence Anisotropy of Lipophilic Probes in Epidermal and Mesophyll Protoplasts from Water-Stressed Lupinus albus L.

The blue emission anisotropy, r, of two lipophilic probes, diphenylhexatriene (DPH) and its trimethyl-ammonium derivative (TMA-DPH), has been measured in foliar Lupinus albus L. protoplasts for the first time by flow cytometry. Distinctive values have been obtained for protoplasts of epidermal and mesophyll origin, identified by their intensities of chlorophyll fluorescence. Fluorescence microscopy confirmed that TMA-DPH remained in the plasma membrane while DPH penetrated into intracellular lipid domains. Typical emission anisotropy values at 22 degrees C for mesophyll and epidermal protoplasts, respectively, were 0.225 and 0.312 with TMA-DPH, and 0.083 and 0.104 with DPH. This indicates that epidermal cells-and notably their plasma membranes (TMA-DPH)-have higher lipid microviscosity and/or more ordered lipid structure. Two lupin genotypes characterized as resistant or susceptible to drought were analyzed with or without 9 days of water stress shown to increase ion leakage from foliar discs. Water stress greatly increased the apparent fluidity, and more so in the susceptible genotype; the effect was more pronounced in the chlorophyll-containing mesophyll cells than in the epidermal cells.

Feasibility of first-pass radionuclide angiocardiography with a 10-mCi technetium bolus using a single-crystal digital gamma camera: implications for technetium-sestamibi single-day protocols.

To test the feasibility of rest first-pass radionuclide angiocardiography (FPRNA) using a 370-MBq (10-mCi) bolus and a single-crystal gamma camera, 40 patients underwent both FPRNA and equilibrium radionuclide angiocardiography (ERNA). Ejection fraction (EF) and regional wall motion (RWM) were assessed by three observers. The interobserver reproducibility was good: the intraclass correlation coefficient was 0.97 for both techniques. The correlation coefficient between FPRNA and ERNA EFs ranged between 0.90 and 0.92. FPRN EFs were significantly higher (P < 0.003) by less than 5 percentage points, this difference having no clinical implications for patient classification. The study provides arguments as to why this difference may be explained self-attenuation rather than counts statistics problems. Both techniques were concordant for RWM analysis. We conclude that FPRNA with 10 mCi is a reliable tool to assess rest left ventricular function, which makes it possible to perform simultaneously myocardial perfusion and function assessment in a single-day protocol using a single-crystal gamma camera.

Catharanthus roseus G-box binding factors 1 and 2 act as repressors of strictosidine synthase gene expression in cell cultures.

The enzyme encoded by the strictosidine synthase (Str) gene catalyses a key step in the biosynthesis of therapeutically valuable terpenoid indole alkaloids. In Catharanthus roseus the Str gene was shown to be regulated by a wide variety of signals including auxin, methyl jasmonate and fungal elicitors in cell suspension cultures and by tissue-specific control in plant organs. The Str promoter contains a functional G-box (CACGTG) cis-regulatory sequence. In order to understand better the mechanisms involved in the regulation of Str gene expression, we isolated the C. roseus cDNAs encoding G-box binding factors Crgbf1 and Crgbf2. The binding specificity of their protein products CrGBF1 and CrGBF2 was analysed by competitive electrophoresis mobility and saturation binding assays. CrGBF1 had a high binding specificity for class I G-boxes including the Str G-box. CrGBF1 showed a lower affinity for class II G-boxes and for the G-box-like element (AACGTG) found in the tryptophan decarboxylase (Tdc) gene which encodes another enzyme involved in TIA biosynthesis. CrGBF2 showed a high affinity for all types of G-boxes tested and to a lesser extent for the Tdc G-box-like element. Transient bombardment experiments demonstrated that both CrGBF1 and CrGBF2 can act in vivo as transcriptional repressors of the Str promoter via direct interaction with the G-box. These data indicate that GBFs may play functional role in the regulation of expression of the terpenoid indole alkaloid biosynthetic gene Str.

Cloning of a cDNA encoding a developmentally regulated 22 kDa polypeptide from tobacco leaf plasma membrane.

A polypeptide doublet (P18-P19, ca 22 kDa, pI 4.5) has been shown to accumulate in tobacco leaf plasma membrane in a development-dependent way, under constant environmental conditions. P18 and P19 were purified by 2D-PAGE and microsequenced. Microsequences revealed only small differences between the two polypeptides. A PCR-based cloning strategy identified a cDNA displaying a 591 bp ORF. The encoded polypeptide contained P19 specific microsequences. It was expressed in E. coli and a specific rabbit antiserum was raised. Western-blots confirmed its identification as P19. The accumulation pattern of hybridizable mRNA around the floral induction period was similar to that of P18 and P19. Searching of databases revealed no significant hits except unidentified plant ESTs. P18 and P19 are proposed as the first example of plant-specific and developmentally regulated plasma membrane proteins.