Dobivanje koncentriranih i pročišćenih fruktooligosaharida iz korijena biljke Smallanthus sonchifolius pomoću membranske tehnologije

Yacon is a perennial plant originating from the Andean region whose roots have been receiving increased att ention due to their high content of prebiotic fructooligosaccharides (FOS). Apart from many health benefi ts, FOS have interesting characteristics as food ingredients, so are used as sugar substitute, and their extraction from yacon roots may be an alternative to commercially available FOS. This work evaluates membrane technology for concentration and purifi cation of FOS from yacon root extract, combining ultrafiltration (UF) with nanofi ltration (NF), with and without the use of discontinuous diafi ltration (DF). After UF, 63.75 % of the saccharides from the initial feed were recovered in total permeate. DF did not largely infl uence FOS retention during NF (it increased from 68.78 % without DF to 70.48 % with DF), but decreased glucose and fructose retentions, from 40.63 to 31.61 % and 25.64 to 18.69 %, respectively, which was desirable, allowing greater purification of FOS in the retentate. The yield of total saccharides in the final retentate after combined UF and NF processes was 50.89 % and of FOS was 51.85 %, with 19.75 % purity. The results indicate that the combined UF and NF is a promising technique for concentrating yacon saccharides, but more diafi ltration steps are required for the improvement of FOS purity.Biljka Smallanthus sonchifolius je trajnica iz područja Anda, čiji je korijen bogat prebiotičkim fruktooligosaharidima, zbog čega je zanimljiva kao sirovina za prehrambenu industriju. Osim što pozitivno utječu na zdravlje, fruktooligosaharidi se zbog svojih svojstava mogu upotrijebiti kao dodatak hrani, te kao zamjena za šećer. Fruktooligosaharidi dobiveni ekstrakcijom iz korijena južnoameričke biljke Smallanthus sonchifolius mogu se upotrijebiti kao zamjena za komercijalno dostupne fruktooligosaharide. U ovom je radu ispitan postupak dobivanja koncentriranih i pročišćenih fruktooligosaharida iz ekstrakta korijena biljke, i to pomoću ultrafiltracije i nanofiltracije, u kombinaciji s diskontinuiranom dijafiltracijom ili bez nje. Nakon ultrafiltracije, 63,75 % se početne količine saharida zadržalo u permeatu. Diskontinuirana dijafiltracija nije bitno utjecala na udjel fruktooligosaharida u retentatu nakon nanofiltracije (bez dijafiltracije udjel je bio 63,78 %, a sa dijafiltracijom 70,48 %), ali je smanjila udjel glukoze, i to s 40,63 na 31,61 %, i fruktoze s 25,64 na 18,69 %, čime je omogućeno bolje pročišćavanje fruktooligosaharida iz retentata. Prinos ukupnih saharida u retentatu nakon kombiniranog tretmana ultrafiltracijom i nanofiltracijom bio je 50,89 %, a fruktooligosaharida 51,85 %, uz čistoću od 19,75 %. Rezultati pokazuju da je kombinacija ultra- i nanofiltracije prikladna metoda dobivanja koncentriranih saharida iz korijena biljke Smallanthus sonchifolius, ali da su za poboljšanje čistoće fruktooligosaharida potrebni su dodatni koraci u postupku dijafiltracije

An estimate of the total number of true human miRNAs

While the number of human miRNA candidates continuously increases, only a few of them are completely characterized and experimentally validated. Toward determining the total number of true miRNAs, we employed a combined in silico high- and experimental low-throughput validation strategy. We collected 28 866 human small RNA sequencing data sets containing 363.7 billion sequencing reads and excluded falsely annotated and low quality data. Our high-throughput analysis identified 65% of 24 127 mature miRNA candidates as likely false-positives. Using northern blotting, we experimentally validated miRBase entries and novel miRNA candidates. By exogenous overexpression of 108 precursors that encode 205 mature miRNAs, we confirmed 68.5% of the miRBase entries with the confirmation rate going up to 94.4% for the high-confidence entries and 18.3% of the novel miRNA candidates. Analyzing endogenous miRNAs, we verified the expression of 8 miRNAs in 12 different human cell lines. In total, we extrapolated 2300 true human mature miRNAs, 1115 of which are currently annotated in miRBase V22. The experimentally validated miRNAs will contribute to revising targetomes hypothesized by utilizing falsely annotated miRNAs

Epstein-Barr Virus EBER Transcripts Affect miRNA-Mediated Regulation of Specific Targets and Are Processed to Small RNA Species

The oncogenic Epstein-Barr virus (EBV) expresses 44 mature microRNAs and two non-coding EBER RNAs of 167 (EBER1) and 172 (EBER2) nt length. MiRNA profiling of NK/T cell lines and primary cells and Northern blotting of EBV-infected cell lines and primary tumors revealed processing of EBER1 to short 5′-derived RNAs of approximately 23, 52 and 70 nt (EBER123, EBER152, and EBER170) and of EBER2 to 3′ fragments. The biogenesis of these species is independent of Dicer, and EBER123 does not act like a miRNA OPEN ACCESS Non-Coding RNA 2015, 1 171 to target its complementary sequence. EBER1, EBER2 and EBER123 were bound by the lupus antigen (La), a nuclear and cytoplasmic protein that facilitates RNAi. Consistent with this, the EBERs affect regulation of interleukin 1alpha (IL1α) and RAC1 reporters harboring miR target sequences, targets of miR-142-3p. However, the EBERs have no effect upon another target of miR-142-3p, ADCY9, nor on TOMM22, a target of ebv-miR-BART16, indicative of selective modulation of gene expression by the EBERs

Large-scale validation of miRNAs by disease association, evolutionary conservation and pathway activity.

The validation of microRNAs (miRNAs) identified by next generation sequencing involves amplification-free and hybridization-based detection of transcripts as criteria for confirming valid miRNAs. Since respective validation is frequently not performed, miRNA repositories likely still contain a substantial fraction of false positive candidates while true miRNAs are not stored in the repositories yet. Especially if downstream analyses are performed with these candidates (e.g. target or pathway prediction), the results may be misleading. In the present study, we evaluated 558 mature miRNAs from miRBase and 1,709 miRNA candidates from next generation sequencing experiments by amplification-free hybridization and investigated their distributions in patients with various disease conditions. Notably, the most significant miRNAs in diseases are often not contained in the miRBase. However, these candidates are evolutionary highly conserved. From the expression patterns, target gene and pathway analyses and evolutionary conservation analyses, we were able to shed light on the complexity of miRNAs in humans. Our data also highlight that a more thorough validation of miRNAs identified by next generation sequencing is required. The results are available in miRCarta ( https://mircarta.cs.uni-saarland.de )

The Proper Motion of the Globular Cluster NGC 6553 and of Bulge Stars with HST

WFPC2 images obtained with the Hubble Space telescope 4.16 years apart have allowed us to measure the proper motion of the metal rich globular cluster NGC 6553 with respect to the background bulge stars. With a space velocity of (${\Pi}, {\Theta}, W$) = (-3.5, 230, -3) km s$^{-1}$, NGC 6553 follows the mean rotation of both disk and bulge stars at a Galactocentric distance of 2.7 kpc. While the kinematics of the cluster is consistent with either a bulge or a disk membership, the virtual identity of its stellar population with that of the bulge cluster NGC6528 makes its bulge membership more likely. The astrometric accuracy is high enough for providing a measure of the bulge proper motion dispersion and confirming its rotation. A selection of stars based on the proper motions produced an extremely well defined cluster color-magnitude diagram (CMD), essencially free of bulge stars. The improved turnoff definition in the decontaminated CMD confirms an old age for the cluster (~13 Gyr) indicating that the bulge underwent a rapid chemical enrichment while being built up at in the early Universe. An additional interesting feature of the cluster color-magnitude diagram is a significant number of blue stragglers stars, whose membership in the cluster is firmly established from their proper motions.Comment: version with full-page figure

ELF5 suppresses estrogen sensitivity and underpins the acquisition of antiestrogen resistance in luminal breast cancer.

We have previously shown that during pregnancy the E-twenty-six (ETS) transcription factor ELF5 directs the differentiation of mammary progenitor cells toward the estrogen receptor (ER)-negative and milk producing cell lineage, raising the possibility that ELF5 may suppress the estrogen sensitivity of breast cancers. To test this we constructed inducible models of ELF5 expression in ER positive luminal breast cancer cells and interrogated them using transcript profiling and chromatin immunoprecipitation of DNA followed by DNA sequencing (ChIP-Seq). ELF5 suppressed ER and FOXA1 expression and broadly suppressed ER-driven patterns of gene expression including sets of genes distinguishing the luminal molecular subtype. Direct transcriptional targets of ELF5, which included FOXA1, EGFR, and MYC, accurately classified a large cohort of breast cancers into their intrinsic molecular subtypes, predicted ER status with high precision, and defined groups with differential prognosis. Knockdown of ELF5 in basal breast cancer cell lines suppressed basal patterns of gene expression and produced a shift in molecular subtype toward the claudin-low and normal-like groups. Luminal breast cancer cells that acquired resistance to the antiestrogen Tamoxifen showed greatly elevated levels of ELF5 and its transcriptional signature, and became dependent on ELF5 for proliferation, compared to the parental cells. Thus ELF5 provides a key transcriptional determinant of breast cancer molecular subtype by suppression of estrogen sensitivity in luminal breast cancer cells and promotion of basal characteristics in basal breast cancer cells, an action that may be utilised to acquire antiestrogen resistance

Analyse nicht-kodierender RNAs in Epstein-Barr Virus infizierten NK/T-Zelllinien

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Synthesis of Purine and Pyrimidine 3'-Amino-3'-deoxy- and 3'-Amino-2',3'-dideoxyxylonucleosides.

International audienceA general procedure to obtain the 3'-aminoxylonucleosides 13a,b and 17a,b is presented. The synthetic scheme is based on the 5' directed intramolecular nucleophilic substitution at the 3'-activated position of the nucleoside. The approach of the incoming group to this position takes place regio- and stereoselectively from the most hindered face of the nucleoside. The methodology presented is applicable to ribonucleosides and 2'-deoxyribonucleosides, regardless of their nitrogenated base