High-confidence glycosome proteome for procyclic form <em>Trypanosoma brucei</em> by epitope-tag organelle enrichment and SILAC proteomics

The glycosome of the pathogenic African trypanosome Trypanosoma brucei is a specialized peroxisome that contains most of the enzymes of glycolysis and several other metabolic and catabolic pathways. The contents and transporters of this membrane-bounded organelle are of considerable interest as potential drug targets. Here we use epitope tagging, magnetic bead enrichment, and SILAC quantitative proteomics to determine a high-confidence glycosome proteome for the procyclic life cycle stage of the parasite using isotope ratios to discriminate glycosomal from mitochondrial and other contaminating proteins. The data confirm the presence of several previously demonstrated and suggested pathways in the organelle and identify previously unanticipated activities, such as protein phosphatases. The implications of the findings are discussed

Three-dimensional visualization software assists learning in students with diverse spatial intelligence in medical education

This study evaluated effect of mental rotation (MR) training on learning outcomes and explored effectiveness of teaching via three-dimensional (3D) software among medical students with diverse spatial intelligence. Data from n = 67 student volunteers were included. A preliminary test was conducted to obtain baseline level of MR competency and was utilized to assign participants to two experimental conditions, i.e., trained group (n = 25) and untrained group (n = 42). Data on the effectiveness of training were collected to measure participants\u27 speed and accuracy in performing various MR activities. Six weeks later, a large class format (LCF) session was conducted for all students using 3D software. The usefulness of technology-assisted learning at the LCF was evaluated via a pre- and post-test. Students\u27 feedback regarding MR training and use of 3D software was acquired through questionnaires. MR scores of the trainees improved from 25.9±4.6 points to 28.1±4.4 (P = 0.011) while time taken to complete the tasks reduced from 20.9±3.9 to 12.2±4.4 minutes. Males scored higher than females in all components (P = 0.016). Further, higher pre- and post-test scores were observed in trained (9.0±1.9 and 12.3±1.6) versus untrained group (7.8±1.8; 10.8±1.8). Although mixed-design analysis of variance suggested significant difference in their test scores (P \u3c 0.001), both groups reported similar trend in improvement by means of 3D software (P = 0.54). Ninety-seven percent of students reported technology-assisted learning as an effective means of instruction and found use of 3D software superior to plastic models. Software based on 3D technologies could be adopted as an effective teaching pedagogy to support learning across students with diverse levels of mental rotation abilities

Enteric dysbiosis and fecal calprotectin expression in premature infants.

BackgroundPremature infants often develop enteric dysbiosis with a preponderance of Gammaproteobacteria, which has been related to adverse clinical outcomes. We investigated the relationship between increasing fecal Gammaproteobacteria and mucosal inflammation, measured by fecal calprotectin (FC).MethodsStool samples were collected from very-low-birth weight (VLBW) infants at ≤2, 3, and 4 weeks' postnatal age. Fecal microbiome was surveyed using polymerase chain reaction amplification of the V4 region of 16S ribosomal RNA, and FC was measured by enzyme immunoassay.ResultsWe enrolled 45 VLBW infants (gestation 27.9 ± 2.2 weeks, birth weight 1126 ± 208 g) and obtained stool samples at 9.9 ± 3, 20.7 ± 4.1, and 29.4 ± 4.9 days. FC was positively correlated with the genus Klebsiella (r = 0.207, p = 0.034) and its dominant amplicon sequence variant (r = 0.290, p = 0.003), but not with the relative abundance of total Gammaproteobacteria. Klebsiella colonized the gut in two distinct patterns: some infants started with low Klebsiella abundance and gained these bacteria over time, whereas others began with very high Klebsiella abundance.ConclusionIn premature infants, FC correlated with relative abundance of a specific pathobiont, Klebsiella, and not with that of the class Gammaproteobacteria. These findings indicate a need to define dysbiosis at genera or higher levels of resolution

The Great Observatories Origins Deep Survey. VLT/FORS2 Spectroscopy in the GOODS-South Field: Part III

Aims. We present the full data set of the spectroscopic campaign of the ESO/GOODS program in the GOODS-South field, obtained with the FORS2 spectrograph at the ESO/VLT. Method. Objects were selected as candidates for VLT/FORS2 observations primarily based on the expectation that the detection and measurement of their spectral features would benefit from the high throughput and spectral resolution of FORS2. The reliability of the redshift estimates is assessed using the redshift-magnitude and color-redshift diagrams, and comparing the results with public data. Results. Including the third part of the spectroscopic campaign (12 masks) to the previous work (26 masks, Vanzella et al. 2005, 2006), 1715 spectra of 1225 individual targets have been analyzed. The actual spectroscopic catalog provides 887 redshift determinations. The typical redshift uncertainty is estimated to be sigma(z) ~ 0.001. Galaxies have been selected adopting different color criteria and using photometric redshifts. The resulting redshift distribution typically spans two domains: from z=0.5 to 2 and z=3.5 to 6.3. The reduced spectra and the derived redshifts are released to the community through the ESO web page http://www.eso.org/science/goods/Comment: 11 pages, 11 figures; accepted for publication in Astronomy & Astrophysics. Data are available at http://www.eso.org/science/goods

Routine Antenatal Anti-D Prophylaxis in Women Who Are Rh(D) Negative: Meta-Analyses Adjusted for Differences in Study Design and Quality

Background: To estimate the effectiveness of routine antenatal anti-D prophylaxis for preventing sensitisation in pregnant Rhesus negative women, and to explore whether this depends on the treatment regimen adopted. Methods: Ten studies identified in a previous systematic literature search were included. Potential sources of bias were systematically identified using bias checklists, and their impact and uncertainty were quantified using expert opinion. Study results were adjusted for biases and combined, first in a random-effects meta-analysis and then in a random-effects metaregression analysis. Results: In a conventional meta-analysis, the pooled odds ratio for sensitisation was estimated as 0.25 (95 % CI 0.18, 0.36), comparing routine antenatal anti-D prophylaxis to control, with some heterogeneity (I 2 = 19%). However, this naïve analysis ignores substantial differences in study quality and design. After adjusting for these, the pooled odds ratio for sensitisation was estimated as 0.31 (95 % CI 0.17, 0.56), with no evidence of heterogeneity (I 2 = 0%). A meta-regression analysis wa

Novel Lipophilic Acetohydroxamic Acid Derivatives Based on Conformationally Constrained Spiro Carbocyclic 2,6-Diketopiperazine Scaffolds with Potent Trypanocidal Activity

We describe novel acetohydroxamic acid derivatives with potent activity against cultured bloodstream-form Trypanosoma brucei and selectivity indices of >1000. These analogues were derived from conformationally constrained, lipophilic, spiro carbocyclic 2,6-diketopiperazine (2,6-DKP) scaffolds by attaching acetohydroxamic acid moieties to the imidic nitrogen. Optimal activity was achieved by placing benzyl groups adjacent to the basic nitrogen of the 2,6-DKP core. S-Enantiomer 7d was the most active derivative against T. brucei (IC(50) = 6.8 nM) and T. cruzi (IC(50) = 0.21 μM)

Women's attitude towards prenatal screening for red blood cell antibodies, other than RhD

Background: Since July 1998 all Dutch women (± 200,000/y) are screened for red cell antibodies, other than anti-RhesusD (RhD) in the first trimester of pregnancy, to facilitate timely treatment of pregnancies at risk for hemolytic disease of the fetus and newborn (HDFN). Evidence for benefits, consequences and costs of screening for non-RhD antibodies is still under discussion. The screening program was evaluated in a nation-wide study. As a part of this evaluation study we investigated, according to the sixth criterium of Wilson and Jüngner, the acceptance by pregnant women of the screening program for non-RhD antibodies. Methods: Controlled longitudinal survey, including a prenatal and a postnatal measurement by structured questionnaires. Main outcome measures: information satisfaction, anxiety during the screening process (a.o. STAI state inventory and specific questionnaire modules), overall attitude on the screening program. Univariate analysis was followed by standard multivariate analysis to identify significant predictors of the outcome measures. Participants: 233 pregnant women, distributed over five groups, according to the screening result. Results: Satisfaction about the provided information was moderate in all groups. All screen- positive groups desired more supportive information. Anxiety increased in screen- positives during the screening process, but decreased to basic levels postnatally. All groups showed a strongly positive balance between perceived utility and burden of the

Genomic and Proteomic Studies on the Mode of Action of Oxaboroles against the African Trypanosome

SCYX-7158, an oxaborole, is currently in Phase I clinical trials for the treatment of human African trypanosomiasis. Here we investigate possible modes of action against Trypanosoma brucei using orthogonal chemo-proteomic and genomic approaches. SILAC-based proteomic studies using an oxaborole analogue immobilised onto a resin was used either in competition with a soluble oxaborole or an immobilised inactive control to identify thirteen proteins common to both strategies. Cell-cycle analysis of cells incubated with sub-lethal concentrations of an oxaborole identified a subtle but significant accumulation of G2 and >G2 cells. Given the possibility of compromised DNA fidelity, we investigated long-term exposure of T. brucei to oxaboroles by generating resistant cell lines in vitro. Resistance proved more difficult to generate than for drugs currently used in the field, and in one of our three cell lines was unstable. Whole-genome sequencing of the resistant cell lines revealed single nucleotide polymorphisms in 66 genes and several large-scale genomic aberrations. The absence of a simple consistent mechanism among resistant cell lines and the diverse list of binding partners from the proteomic studies suggest a degree of polypharmacology that should reduce the risk of resistance to this compound class emerging in the field. The combined genetic and chemical biology approaches have provided lists of candidates to be investigated for more detailed information on the mode of action of this promising new drug clas