Structure of the exon junction core complex with a trapped DEAD-box ATPase bound to RNA

In higher eukaryotes, a multiprotein exon junction complex is deposited on spliced messenger RNAs. The complex is organized around a stable core, which serves as a binding platform for numerous factors that influence messenger RNA function. Here, we present the crystal structure of a tetrameric exon junction core complex containing the DEAD-box adenosine triphosphatase (ATPase) eukaryotic initiation factor 4AIII (eIF4AIII) bound to an ATP analog, MAGOH, Y14, a fragment of MLN51, and a polyuracil mRNA mimic. eIF4AIII interacts with the phosphate-ribose backbone of six consecutive nucleotides and prevents part of the bound RNA from being double stranded. The MAGOH and Y14 subunits lock eIF4AIII in a prehydrolysis state, and activation of the ATPase probably requires only modest conformational changes in eIF4AIII motif I.     

Ontogeny of T cells in the pig fetus

Mononuclear cells isolated from thymus, spleen and cord blood of pig fetuses ranging in age from 48 to 112 days were examined for the presence of sheep red blood cell rosette-forming cells (SRBC-RFC). After an initial increase from 77 % (mean) at 48 days of gestation to 88 % at 60 days, the proportion of SRBC-RFC in thymus remained constant throughout the gestational period. In spleen and cord blood, the proportion of SRBC-RFC increased with age, from occasional rosette-forming cells at 48 days of gestation to 21 % and 30 %, respectively, at 112 days. The demonstrated development of SRBC-RFC in the thymus, spleen and cord blood is considered to reflect the ontogeny of T cells in these fetal pig tissues.     

Accurate Dereplication of Bioactive Secondary Metabolites from Marine-Derived Fungi by UHPLC-DAD-QTOFMS and a MS/HRMS Library

In drug discovery, reliable and fast dereplication of known compounds is essential for identification of novel bioactive compounds. Here, we show an integrated approach using ultra-high performance liquid chromatography-diode array detection-quadrupole time of flight mass spectrometry (UHPLC-DAD-QTOFMS) providing both accurate mass full-scan mass spectrometry (MS) and tandem high resolution MS (MS/HRMS) data. The methodology was demonstrated on compounds from bioactive marine-derived strains of Aspergillus, Penicillium, and Emericellopsis, including small polyketides, non-ribosomal peptides, terpenes, and meroterpenoids. The MS/HRMS data were then searched against an in-house MS/HRMS library of ~1300 compounds for unambiguous identification. The full scan MS data was used for dereplication of compounds not in the MS/HRMS library, combined with ultraviolet/visual (UV/Vis) and MS/HRMS data for faster exclusion of database search results. This led to the identification of four novel isomers of the known anticancer compound, asperphenamate. Except for very low intensity peaks, no false negatives were found using the MS/HRMS approach, which proved to be robust against poor data quality caused by system overload or loss of lock-mass. Only for small polyketides, like patulin, were both retention time and UV/Vis spectra necessary for unambiguous identification. For the ophiobolin family with many structurally similar analogues partly co-eluting, the peaks could be assigned correctly by combining MS/HRMS data and m/z of the [M + Na]⁺ ions.     

Incidence, appearance and development of potato mop-top furovirus-induced spraing in potato cultivars and the influence on yield, distribution in Denmark and detection of the virus in tubers by ELISA

Summary 23 potato cultivars were grown in fields infested with potato mop-top virus and spraing was assessed on tubers at harvest and after storage. Large differences in spraing susceptibility were found between cultivars. Compared with other countries, spraing in Denmark is characterised by a high incidence at harvest, a low increase during storage and a very low proportion of superficial spraing. Decrease in spraing during storage was recorded in two cultivars. DASELISA tests for PMTV in tubers revealted a high reliability. Mapping of PMTV in important Danish potato growing areas showed that the virus is widespread. Occurrence of spraing did not influence total yield or dry matter content. Soil acidity did not influence incidence of spraing but it was more common on coarse-grained soil than on finer sandy soils. PMTV in viruliferous resting spores ofSpongospora subterranea was inactivated by heating to 90°C for 15 minutes.     

The degree of starch phosphorylation as influenced by phosphate deprivation of potato (Solanum tuberosum L.) plants

Summary The levels of starch phosphorylation in potato tubers were investigated in relation to fertilization with phosphorus (P) of field-grown and greenhouse-grown plants. The field-grown plants received 0, 15 or 30 kg P ha⁻¹. Starch from plants grown without P-fertilizer contained 15.6 nmol P (mg starch)⁻¹ whereas starch from plants grown with 30 kg P ha⁻¹ contained 20.6 nmol P (mg starch)⁻¹. The greenhouse-grown plants were cultured in inert media and received nutrient solutions containing 0, 3, 12 or 24 mg P l⁻¹, respectively. Plants grown with no P-fertilizer produced tuber starch with phosphorylation levels reduced to approximately 30% of the level found in plants grown with an ample supply of P. Thus, the level of starch phosphorylation can be modified by limiting the P-supply through the root system. Application of foliar P-fertilizer to the greenhouse-grown plants had no significant effect on the phosphorylation of the tuber starch.     

The Effect of Low-Dose Marine n-3 Fatty Acids on Plasma Levels of sCD36 in Overweight Subjects: A Randomized,Double-Blind,Placebo-Controlled Trial

CD36 is a scavenger receptor involved in lipid uptake and inflammation. Recently, non-cell-bound CD36 (sCD36) was identified in plasma and suggested to be a marker of lipid accumulation in the vessel wall. Marine n-3 polyunsaturated fatty acids (PUFA) may have cardioprotective effects. This study evaluated the effect of marine n-3 PUFA on sCD36 levels in overweight subjects. Fifty overweight subjects were randomized to 1.1 g of n-3 PUFA or 2 g of olive oil daily for six weeks. Neutrophils were isolated at baseline and after six weeks of treatment while an adipose tissue biopsy was obtained at baseline. The content of n-3 PUFA in adipose tissue and neutrophils was analyzed by gas chromatography, while plasma levels of sCD36 were determined using an enzyme-linked immunosorbent assay (ELISA). After six weeks of supplement plasma sCD36 did not differ between supplements (P = 0.18). There was no significant correlation between plasma sCD36 levels and n-3 PUFA in neutrophils at baseline (r = −0.02, P = 0.88), after six weeks supplement (r = −0.03, P = 0.85) or in adipose tissue (r = 0.14, P = 0.34). This study therefore does not provide evidence for a cardioprotective effect of n-3 PUFA acting through a CD36-dependent mechanism.