Cell Wall Epitopes and Endoploidy as Reporters of Embryogenic Potential in Brachypodium Distachyon Callus Culture

Effective regeneration of callus tissue into embryos and then into whole plants is essential for plant biotechnology. The embryonic potential is often low and can further decrease with time in culture, which limits the utilisation of calli for transformation procedures and in vitro propagation. In this study, we show that the loss of embryogenic potential in callus cultures of Brachypodium distachyon is progressive over time. Flow cytometry analyses indicated endoploidy levels increased in 60- and 90-day-old calli with effective loss of the 2C DNA content peak in the latter. Analysis of indolic compounds content revealed a decrease in 60- and 90-day-old calli compared to either freshly isolated explants or 30-day-old calli. Immunohistochemical analysis revealed a decrease in arabinogalactan proteins (AGP) signal with the time of culture, but extensin (EXT) epitopes either increased (JIM12 epitopes) or decreased (JIM11 epitopes). The transcript accumulation levels of AGPs and EXTs confirmed these results, with most of AGP and EXT transcripts gradually decreasing. Some chimeric EXT transcripts significantly increased on the 30th day of culture, perhaps because of an increased embryogenic potential. Selected somatic embryogenesis-related genes and cyclins demonstrated a gradual decrease of transcript accumulation for YUCCA (YUC), AINTEGUMENTA-LIKE (AIL), BABY BOOM (BBM), and CLAVATA (CLV3) genes, as well as for most of the cyclins, starting from the 30th day of culture. Notably, WUSCHEL (WUS) transcript was detectable only on the 30th and 60th day and was not detectable in the zygotic embryos and in 90-day-old calli

LSST Cadence Optimization White Paper in Support of Observations of Unresolved Tidal Stellar Streams in Galaxies beyond the Local Group

Deep observations of faint surface brightness stellar tidal streams in external galaxies with LSST are addressed in this White Paper contribution. We propose using the Wide--Fast--Deep survey that contains several nearby galaxies (at distances where the stars themselves are not resolved, i.e., beyond 20 Mpc). In the context of hierarchical galaxy formation, it is necessary to understand the prevalence and properties of tidal substructure around external galaxies based on integrated (i.e., unresolved) diffuse light. This requires collecting observations on much larger samples of galaxies than the Milky Way and M31. We will compare the observed structures to the predictions of cosmological models of galactic halo formation that inform us about the number and properties of streams around Milky Way-like galaxies. The insight gained from these comparisons will allow us to infer the properties of stream progenitors (masses, dynamics, metallicities, stellar populations). The changes in the host galaxies caused by the interactions with the dissolving companion galaxies will be another focus of our studies. We conclude by discussing synergies with WFIRST and Euclid, and also provide concrete suggestions for how the effects of scattered light could be minimized in LSST images to optimize the search for low surface brightness features, such as faint unresolved stellar tidal streams.Comment: 13 pages, 2 figures, submitted to Call for White Papers on LSST Cadence Optimizatio

Survey of hormone activities in municipal biosolids and animal manures.

ABSTRACT: The potential exists for natural or synthetic hormonal chemicals present in agricultural fertilizers to be transferred to adjacent aquatic environments in order to alter endocrine function in exposed wildlife. Recombinant yeast and mammalian cell line (BG1Luc4E 2 ) assays were used to screen crude organic extracts of municipal biosolids and animal manures for estrogen-, androgen-, and progesterone receptor gene transcription activities. Of the biosolid extracts, those samples that had undergone aerobic digestion had no or minimal estrogen-and no androgen receptor gene transcription activities. In contrast, those biosolid samples that had undergone anaerobic digestion had much higher estrogen-and, for all but one site, androgen receptor gene transcription activities. Extracts prepared from animal manure samples had variable levels of androgen-and estrogen receptor gene transcription activities, which may be related to the type, sex, age, and reproductive status of the animals. The diet and treatment of animals with hormone implants also appeared to be factors influencing hormone activity in animal manure. Progesterone receptor gene transcription activity was observed for only one chicken litter sample. Overall, results of this study suggest that in vitro bioassays can be used to survey and detect hormone activity in municipal biosolids and animal manures. Furthermore, results of these assays can be used to develop 216 practices that will minimize the potential environmental endocrine-disrupting effects of these substances

Routine clinical cardiovascular magnetic resonance in paediatric and adult congenital heart disease: patients, protocols, questions asked and contributions made

<p>Abstract</p> <p>Background</p> <p>Cardiovascular Magnetic Resonance (CMR) of patients with congenital heart disease (CHD) has become routine clinical practice. However, existing CMR protocols focus predominantly on patients with ischemic heart disease, and information is limited on the types of patient with CHD who benefit from CMR investigation, and in what ways. Therefore the aim of this study was to answer the questions: What type of patients were studied by CMR in a centre specializing in paediatric and adult CHD management? What questions were asked, which protocols were used and were the questions successfully answered? To answer these questions, we conducted a cohort study of all 362 patients that received routine clinical CMR during 2007 at the Department of Paediatric Cardiology and Congenital Heart Disease at the Deutsches Herzzentrum München.</p> <p>Results</p> <p>Underlying diagnosis was in 33% Fallot's tetralogy, 17% aortic coarctation, 8% Ebstein's disease, 6% Marfan's disease, 4% single ventricle with Fontan-like circulation, and 32% others. Median age was 26 years (7 days – 75 years). Ventricular volumes were assessed in 67% of the patients; flow in 74%; unknown anatomy only in 9%; specific individual morphology of known anatomy in 83%; myocardial fibrosis in 8%; stress-induced myocardial perfusion defects in 1%. Only in 3% of the cases the question could not be fully answered.</p> <p>Conclusion</p> <p>Contrary to common belief, routine CMR of patients with CHD was not requested to address global anatomical questions so much as to clarify specific questions of morphology and function of known anatomy. The CMR protocols used differed markedly from those widely used in patients with ischemic heart disease.</p

Individual component analysis of the multi-parametric cardiovascular magnetic resonance protocol in the CE-MARC trial

Background: The CE-MARC study assessed the diagnostic performance investigated the use of cardiovascular magnetic resonance (CMR) in patients with suspected coronary artery disease (CAD). The study used a multi-parametric CMR protocol assessing 4 components: i) left ventricular function; ii) myocardial perfusion; iii) viability (late gadolinium enhancement (LGE)) and iv) coronary magnetic resonance angiography (MRA). In this pre-specified CE-MARC sub-study we assessed the diagnostic accuracy of the individual CMR components and their combinations. Methods: All patients from the CE-MARC population (n = 752) were included using data from the original blinded-read. The four individual core components of the CMR protocol was determined separately and then in paired and triplet combinations. Results were then compared to the full multi-parametric protocol. Results: CMR and X-ray angiography results were available in 676 patients. The maximum sensitivity for the detection of significant CAD by CMR was achieved when all four components were used (86.5 %). Specificity of perfusion (91.8 %), function (93.7 %) and LGE (95.8 %) on its own was significantly better than specificity of the multi-parametric protocol (83.4 %) (all P < 0.0001) but with the penalty of decreased sensitivity (86.5 % vs. 76.9 %, 47.4 % and 40.8 % respectively). The full multi-parametric protocol was the optimum to rule-out significant CAD (Likelihood Ratio negative (LR-) 0.16) and the LGE component alone was the best to rue-in CAD (LR+ 9.81). Overall diagnostic accuracy was similar with the full multi-parametric protocol (85.9 %) compared to paired and triplet combinations. The use of coronary MRA within the full multi-parametric protocol had no additional diagnostic benefit compared to the perfusion/function/LGE combination (overall accuracy 84.6 % vs. 84.2 % (P = 0.5316); LR- 0.16 vs. 0.21; LR+ 5.21 vs. 5.77). Conclusions: From this pre-specified sub-analysis of the CE-MARC study, the full multi-parametric protocol had the highest sensitivity and was the optimal approach to rule-out significant CAD. The LGE component alone was the optimal rule-in strategy. Finally the inclusion of coronary MRA provided no additional benefit when compared to the combination of perfusion/function/LGE. Trial registration: Current Controlled Trials ISRCTN77246133