3D ultrastructural organization of whole Chlamydomonas reinhardtii cells studied by nanoscale soft x-ray tomography

The complex architecture of their structural elements and compartments is a hallmark of eukaryotic cells. The creation of high resolution models of whole cells has been limited by the relatively low resolution of conventional light microscopes and the requirement for ultrathin sections in transmission electron microscopy. We used soft x-ray tomography to study the 3D ultrastructural organization of whole cells of the unicellular green alga Chlamydomonas reinhardtii at unprecedented spatial resolution. Intact frozen hydrated cells were imaged using the natural x-ray absorption contrast of the sample without any staining. We applied different fiducial-based and fiducial-less alignment procedures for the 3D reconstructions. The reconstructed 3D volumes of the cells show features down to 30 nm in size. The whole cell tomograms reveal ultrastructural details such as nuclear envelope membranes, thylakoids, basal apparatus, and flagellar microtubule doublets. In addition, the x-ray tomograms provide quantitative data from the cell architecture. Therefore, nanoscale soft x-ray tomography is a new valuable tool for numerous qualitative and quantitative applications in plant cell biology

Course and Outcome of Bacteremia Due to Staphylococcus Aureus: Evaluation of Different Clinical Case Definitions

In a retrospective survey of patients hospitalized in the University Hospital of Basel, Switzerland, the course and outcome of 281 cases of true bacteremia due to Staphylococcus aureus over a 7-year period were analyzed. The main purpose was to evaluate different case definitions. In 78% of cases the source of bacteremia was obvious; vascular access sites (27%) and wounds (10%) were the most common sources. Metastasizing foci were more common in cases of primary vs. secondary bacteremia (P <.001). The incidence of endocarditis was higher in cases in which no portal of entry was defined (P <.03). The overall mortality rate was high at 34% partly because of inappropriate initial antibiotic therapy. With the introduction of an infectious disease service at the hospital, the fraction of misjudged results of blood culture diminished 2.5-fold. Among the differently defined cases, the mortality rate was significantly higher for cases of complicated vs. uncomplicated bacteremia (P <.01), for cases of primary vs. secondary bacteremia (P = .05), and for patients with endocarditis or other secondary foci (P <.001). Since only one methicillin-resistant strain was isolated, multiresistant staphylococci were not a problem in the hospital. Different case definitions allowed the detection of patients at increased risk for complications and death. In the treatment of sepsis with no evident focus, initial antimicrobial therapy should include the use of agents with antistaphylococcal activity

Partitioning carbon fluxes in a Mediterranean oak forest to disentangle changes in ecosystem sink strenght during drought

Net carbon flux partitioning was used to disentangle abiotic and biotic drivers of all important component fluxes influencing the overall sink strength of a Mediterranean ecosystem during a rapid spring to summer transition. Between May and June 2006 we analyzed how seasonal drought affected ecosystem assimilation and respiration fluxes in an evergreen oak woodland and attributed variations in the component fluxes (trees, understory, soil microorganisms and roots) to observations at the ecosystem scale. We observed a two thirds decrease in both ecosystem carbon assimilation and respiration (Reco) within only 15 days time. The impact of decreasing Reco on the ecosystem carbon balance was smaller than the impact of decreasing primary productivity. Flux partitioning of GPP and Reco into their component fluxes from trees, understory, soil microorganisms and roots showed that declining ecosystem sink strength was due to a large drought and temperatureinduced decrease in understory carbon uptake (from 56% to 21%). Hence, the shallow-rooted annuals mainly composing the understory have a surprisingly large impact on the source/ sink behavior of this open evergreen oak woodland during spring to summer transition and the timing of the onset of drought might have a large effect on the annual carbon budget. In response to seasonal drought Reco was increasingly dominated by respiration of heterotrophic soil microorganisms, while the root flux was found to be of minor importance. Soil respiration flux decreased with drought but its contribution to total daily CO2-exchange increased by 11.5%. This partitioning approach disentangled changes in respiratory and photosynthetic ecosystem fluxes that were not apparent from the eddy-covariance or the soil respiration data alone. By the novel combination of understory vs. overstory carbon flux partitioning with soil respiration data fromtrenched and control plots, we gained a detailed understanding of factors controlling net carbon exchange of Mediterranean ecosystem

Evaluation of non-chemical seed treatment methods for the control of Alternaria dauci and A. radicina on carrot seeds

The current study was initiated to evaluate the efficacy of physical methods (hot water, aerated steam, electron treatment) and agents of natural origin (resistance inducers, plant derived products, micro-organisms) as seed treatments of carrots for control of Alternaria dauci and A. radicina. Control of both Alternaria species by seed treatment with the resistance inducers was generally poor. Results were also not satisfactory with most of the formulated commercial micro-organism preparations. Based on the average of five field trials, one of these, BA 2552 (Pseudomonas chlororaphis), provided a low but significant increase in plant stand. Among the experimental micro-organisms, the best results were obtained with Pseudomonas sp. strain MF 416 and Clonostachys rosea strain IK726. A similar level of efficacy was provided by seed treatment with an emulsion (1%) of thyme oil in water. Good and consistent control was generally achieved with the physical methods aerated steam, hot water and electron treatment. Aerated steam treatment was, apart from the thiram-containing chemical standard, the best single treatment, and its performance may at least partially be due to extensive pre-testing, resulting in dosages optimally adapted to the respective seed lot. In some of the experiments the effect of the hot water treatment, which was tested at a fixed, not specifically adapted dosage, was significantly improved when combined with a Pseudomonas sp. MF 416 or C. rosea IK726 treatment. The results are discussed in relation to the outcome of experiments in which the same seed treatment methods and agents were tested in other seed-borne vegetable pathosystems

Paired EMI-HIMU hotspots in the South Atlantic-Starting plume heads trigger compositionally distinct secondary plumes?

Age-progressive volcanism is generally accepted as the surface expression of deep-rooted mantle plumes, which are enigmatically linked with the African and Pacific large low-shear velocity provinces (LLSVPs). We present geochemical and geochronological data collected from the oldest portions of the age-progressive enriched mantle one (EMI)-type Tristan-Gough track. They are part of a 30- to 40-million year younger age-progressive hotspot track with St. Helena HIMU (high time-integrated U-238/Pb-204) composition, which is also observed at the EMI-type Shona hotspot track in the southernmost Atlantic. Whereas the primary EMI-type hotspots overlie the margin of the African LLSVP, the HIMU-type hotspots are located above a central portion of the African LLSVP, reflecting a large-scale geochemical zonation. We propose that extraction of large volumes of EMI-type mantle from the margin of the LLSVP by primary plume heads triggered upwelling of HIMU material from a more internal domain of the LLSVP, forming secondary plumes

A progressive increase in cardiovascular risk assessed by coronary angiography in non-diabetic patients at sub-diabetic glucose levels

<p>Abstract</p> <p>Objective</p> <p>Diabetes mellitus type 2 (DM2) is a risk factor for coronary heart disease (CHD). While there is a clear correlation of fasting blood glucose (FBG) and 2 h post-challenge blood glucose values (2h-BG) with microvascular complications, the risk for CHD conferred by glucose dysregulation antecedent to DM2 is less clear. Therefore, we investigated associations of FBG and 2h-BG values with the prevalence of CHD assessed by coronary angiography as the most sensitive diagnostic tool.</p> <p>Research Design and Methods</p> <p>Coronary angiography was performed in 1394 patients without known DM. Capillary blood glucose was analyzed before and 2 h after an oral glucose tolerance test. Associations between FBG as well as 2h-BG levels and the risk for CHD were assessed by logistic regression analysis.</p> <p>Results</p> <p>1064 (75%) of patients were diagnosed with CHD. 204 (15%) were diagnosed with so far unknown DM2, 274 (20%) with isolated impaired fasting glucose (IFG), 188 (13%) with isolated impaired glucose tolerance (IGT) and 282 (20%) with both, IGT and IFG. We found a continuous increase in the risk for CHD with fasting and post-challenge blood glucose values even in the subdiabetic range. This correlation did however not suggest clear cut-off values. The increase in risk for CHD reached statistical significance at FBG levels of > 120 mg/dl (Odds Ratio of 2.7 [1.3-5.6] and 2h-BG levels > 140 mg/dl (141-160 mg/dl OR 1.8 [1.1-2.9], which was however lost after adjusting for age, sex and BMI.</p> <p>Conclusions</p> <p>In our study population we found a continuous increased risk for CHD at fasting and 2h-BG levels in the sub-diabetic glucose range, but no clear cut-off values for cardiovascular risk.</p

Discovery of Sexual Dimorphisms in Metabolic and Genetic Biomarkers

Metabolomic profiling and the integration of whole-genome genetic association data has proven to be a powerful tool to comprehensively explore gene regulatory networks and to investigate the effects of genetic variation at the molecular level. Serum metabolite concentrations allow a direct readout of biological processes, and association of specific metabolomic signatures with complex diseases such as Alzheimer's disease and cardiovascular and metabolic disorders has been shown. There are well-known correlations between sex and the incidence, prevalence, age of onset, symptoms, and severity of a disease, as well as the reaction to drugs. However, most of the studies published so far did not consider the role of sexual dimorphism and did not analyse their data stratified by gender. This study investigated sex-specific differences of serum metabolite concentrations and their underlying genetic determination. For discovery and replication we used more than 3,300 independent individuals from KORA F3 and F4 with metabolite measurements of 131 metabolites, including amino acids, phosphatidylcholines, sphingomyelins, acylcarnitines, and C6-sugars. A linear regression approach revealed significant concentration differences between males and females for 102 out of 131 metabolites (p-values<3.8 x 10(-4); Bonferroni-corrected threshold). Sex-specific genome-wide association studies (GWAS) showed genome-wide significant differences in beta-estimates for SNPs in the CPS1 locus (carbamoyl-phosphate synthase 1, significance level: p<3.8 x 10(-10); Bonferroni-corrected threshold) for glycine. We showed that the metabolite profiles of males and females are significantly different and, furthermore, that specific genetic variants in metabolism-related genes depict sexual dimorphism. Our study provides new important insights into sex-specific differences of cell regulatory processes and underscores that studies should consider sex-specific effects in design and interpretation

The bright blue side of the night sky: Spectroscopic survey of bright and hot (pre-) white dwarfs

We report on the spectroscopic confirmation of 68 new bright ($G=13.5-17.2$ mag) and blue (pre-)white dwarfs (WDs). This finding has allowed us to almost double the number of the hottest ($T_{\mathrm{eff}} \geq 60$kK) known WDs brighter than $G=16$ mag. We increased the number of known ultra-high excitation (UHE) WDs by 20%, found one unambiguous close binary system consisting of one DA WD with an irradiated low-mass companion, one DAO, and one DOA WD that are likely in their transformation phase of becoming pure DA WDs, one rare, naked O(H) star, two DA and two DAO WDs with $T_{\mathrm{eff}}$ possibly in excess of 100kK, three new DOZ WDs, and three of our targets are central stars of (possible) planetary nebulae. Using non-local thermodynamic equilibrium models, we derived the atmospheric parameters of these stars and by fitting their spectral energy distribution we derived their radii, luminosities, and gravity masses. In addition, we derived their masses in the Kiel and Hertzsprung-Russell diagram (HRD). We find that Kiel, HRD, and gravity mass agree only in half of the cases. This is not unexpected and we attribute this to the neglect of metal opacities, possibly stratified atmospheres, as well as possible uncertainties of the parallax zero point determination. Furthermore, we carried out a search for photometric variability in our targets using archival data, finding that 26% of our targets are variable. This includes 15 new variable stars, with only one of them being clearly an irradiation effect system. Strikingly, the majority of the variable stars exhibit non-sinusoidal light-curve shapes, which are unlikely explained in terms of close binary systems. We propose that a significant fraction of all (not just UHE) WDs develop spots when entering the WD cooling phase. We suggest that this could be related to the on-set of weak magnetic fields and possibly diffusion.Comment: Accepted for publication in A&

Iron Status and Analysis of Efficacy and Safety of Ferric Carboxymaltose Treatment in Patients with Inflammatory Bowel Disease

Background and Aims:We analyzed iron deficiency and the therapeutic response following intravenous ferric carboxymaltose in a large single-center inflammatory bowel disease (IBD) cohort. Methods: 250 IBD patients were retrospectively analyzed for iron deficiency and iron deficiency anemia. A subgroup was analyzed regarding efficacy and side effects of iron supplementation with ferric carboxymaltose. Results: In the cohort (n = 250), 54.4% of the patients had serum iron levels 60 mu g/dl, 61.6% had ferritin >100 ng/ml, and 90.7% reached Hb >12/13 g/dl at follow-up (p < 0.0001 for all parameters vs. pretreatment values). The most frequent adverse event was a transient increase of liver enzymes with male gender as risk factor (p = 0.008, OR 8.62, 95% CI 1.74-41.66). Conclusions: Iron deficiency and anemia are frequent in IBD patients. Treatment with ferric carboxymaltose is efficious, safe and well tolerated in iron-deficient IBD patients. Copyright (C) 2011 S. Karger AG, Base

Single molecule MATAC-seq reveals key determinants of DNA replication origin efficiency

Stochastic origin activation gives rise to significant cell-to-cell variability in the pattern of genome replication. The molecular basis for heterogeneity in efficiency and timing of individual origins is a long-standing question. Here, we developed Methylation Accessibility of TArgeted Chromatin domain Sequencing (MATAC-Seq) to determine single-molecule chromatin accessibility of four specific genomic loci. MATAC-Seq relies on preferential modification of accessible DNA by methyltransferases combined with Nanopore-Sequencing for direct readout of methylated DNA-bases. Applying MATAC-Seq to selected early-efficient and late-inefficient yeast replication origins revealed large heterogeneity of chromatin states. Disruption of INO80 or ISW2 chromatin remodeling complexes leads to changes at individual nucleosomal positions that correlate with changes in their replication efficiency. We found a chromatin state with an accessible nucleosome-free region in combination with well-positioned +1 and +2 nucleosomes as a strong predictor for efficient origin activation. Thus, MATAC-Seq identifies the large spectrum of alternative chromatin states that co-exist on a given locus previously masked in population-based experiments and provides a mechanistic basis for origin activation heterogeneity during eukaryotic DNA replication. Consequently, our single-molecule chromatin accessibility assay will be ideal to define single-molecule heterogeneity across many fundamental biological processes such as transcription, replication, or DNA repair in vitro and ex vivo