HbA 1c is the stable glucose adduct to N-terminal group of β-chain 0 . The measurement in human blood most important for long-term control glycaemic state diabetic patients. Because there was no internationally agreed reference method IFCC Working Group on Standardization developed a which here described. In first step haemoglobin cleaved into peptides by enzyme endoproteinase Glu-C, and second glycated non-glycated hexapeptides obtained are separated quantified HPLC electrospray ionisation...

A reference method that specifically measures hemoglobin (Hb) A1c is an essential part of the system for international standardization Hb A1c/glycohemoglobin. We have developed a new quantification, based on specific N-terminal residue beta-chains. Enzymatic cleavage intact molecule with endoproteinase Glu-C has been optimized to obtain beta-N-terminal hexapeptides and A0. These peptides separated by reversed-phase HPLC quantitated electrospray ionization-mass spectrometry (method A) or...

Methylation of 3'-terminal nucleotides miRNA/miRNA* is part miRNAs biogenesis in plants but not found animals. In Arabidopsis thaliana this reaction carried out by a multidomain AdoMet-dependent 2'-O-methyltransferase HEN1. Using deletion and structure-guided mutational analysis, we show that the double-stranded RNA-binding domains R(1) R(2) HEN1 make significant uneven contributions to substrate RNA binding, map residues each domain responsible for function. GST pull-down assays yeast...

The complex nature of crop genomes has long prohibited the efficient isolation agronomically relevant genes. However, recent advances in next-generation sequencing technologies provide new ways to accelerate fine-mapping and gene crops. We used RNA allelic six-rowed spike3 (vrs3) mutants with altered spikelet development for identification functional analysis barley (Hordeum vulgare). Variant calling two vrs3 revealed that VRS3 encodes a putative histone Lys demethylase conserved zinc finger...

Abstract We prepared a candidate primary reference material for the forthcoming international standardisation of β-N-terminal glycated hemoglobin A measurements. It consists well-defined mixtures purified and non-glycated A. First, were isolated, to homogeneity, characterised. The techniques used cation exchange affinity chromatography purification, high performance liquid chromatography, capillary isoelectric focusing, electrospray ionisation mass spectrometry, peptide mapping...

DNA damage repair is an essential cellular mechanism that maintains genome stability. Here, we show the nonmethylable cytidine analog zebularine induces a response in Arabidopsis thaliana, independent of changes methylation. In contrast to genotoxic agents induce cell cycle stage-independent manner, specifically during strand synthesis replication. The signaling this mediated by additive activity ATAXIA TELANGIECTASIA MUTATED AND RAD3-RELATED and kinases, which cause postreplicative arrest...

Summary Repetitive DNA sequences and some genes are epigenetically repressed by transcriptional gene silencing (TGS). When genetic mutants not available or problematic to use, TGS can be suppressed chemical inhibitors. However, informed use of epigenetic inhibitors is partially hampered the absence any systematic comparison. In addition, there emerging evidence that cause genomic instability, but nature this damage its repair remain unclear. To bridge these gaps, we compared effects...

Biosynthesis of the capsular K5 polysaccharide Escherichia coli, which has structure 4)-beta GlcA-1,4-alpha GlcNAc-(1, was studied with membrane preparations from an E. coli wild-type strain and a recombinant K-12 expressing capsule. Polymerization occurs at inner face cytoplasmic without participation lipid-linked oligosaccharides. The serological specificity in vitro product determined K5-specific monoclonal antibody antigen-binding assay. polysaccharide, as obtained membranes after...

In plants, a particular class of short interfering (si)RNAs can serve as signal to induce cytosine methylation at homologous genomic regions. If the targeted DNA has promoter function, this RNA-directed (RdDM) result in transcriptional gene silencing (TGS). (RdTGS) transgenes provides versatile system for study epigenetic regulation. We used transcription nopaline synthase (ProNOS)-inverted repeat (IR) provide RNA that triggers de novo and TGS ProNOS copy trans. Utilizing ProNOS-NPTII...

The activity of the cytoplasmic CMP-2-keto-3-deoxyoctulosonic acid synthetase (CMP-KDO synthetase), which is low in Escherichia coli rough strains such as E. K-12 and uncapsulated O111, was significantly elevated encapsulated O10:K5 O18:K5. This enzyme even higher an clone expressing K5 capsule. following findings suggest a correlation between CMP-KDO biosynthesis capsular polysaccharide. (i) Expression polysaccharide were observed with bacteria grown at 37 degrees C but not cells 20 or...

The temperature-regulated expression of capsular group II polysaccharides Escherichia coli (B. Jann and K. Jann, (1990) Curr. Top. Microbiol. Immunol. 150: 19–42) depends on an elevated concentration CMP-KDO, as evidenced by increased activity CMP-KDO synthetase. increase in synthetase is observed only cytoplasmic fractions bacteria which had been grown at 37°C but not after growth 18°C. thus parallels the (membrane-associated) system synthesizing capsules E. coli. No such dependence capsule...

The temperature-regulated expression of capsular group II polysaccharides Escherichia coli (B. Jann and K. Jann, (1990) Curr. Top. Microbiol. Immunol. 150: 19–42) depends on an elevated concentration CMP-KDO, as evidenced by increased activity CMP-KDO synthetase. increase in synthetase is observed only cytoplasmic fractions bacteria which had been grown at 37°C but not after growth 18°C. thus parallels the (membrane-associated) system synthesizing capsules E. coli. No such dependence capsule...

Summary In plants, 24 nucleotide short interfering RNA s serve as a signal to direct cytosine methylation at homologous DNA regions in the nucleus. If targeted has promoter function, this ‐directed may result transcriptional gene silencing. genetic screen for factors involved silencing of Pro NOS – NPTII reporter transgene Arabidopsis thaliana , we captured alleles DOMAINS REARRANGED METHYLTRANSFERASE 2 encoding methyltransferase that is mainly responsible de novo context methylation....

Abstract DNA–protein cross-links (DPCs) are highly toxic DNA lesions consisting of proteins covalently attached to chromosomal DNA. Unrepaired DPCs physically block replication and transcription. Three DPC repair pathways have been identified in Arabidopsis (Arabidopsis thaliana) date: the endonucleolytic cleavage by structure-specific endonuclease MUS81; proteolytic degradation crosslinked protein metalloprotease WSS1A; cross-link phosphodiester bonds tyrosyl phosphodiesterases TDP1 TDP2....

Abstract Loss of genome stability leads to reduced fitness, fertility and a high mutation rate. Therefore, the is guarded by pathways monitoring its integrity neutralizing DNA lesions. To analyze mechanism damage induction cytidine analog zebularine, we performed forward-directed suppressor genetic screen in background Arabidopsis thaliana zebularine-hypersensitive structural maintenance chromosomes 6b (smc6b) mutant. We show that smc6b hypersensitivity was suppressed mutations EQUILIBRATIVE...

Summary Repetitive sequences are ubiquitous components of all eukaryotic genomes. They contribute to genome evolution and the regulation gene transcription. However, uncontrolled activity repetitive can negatively affect functions stability. Therefore, DNA s embedded in a highly repressive heterochromatic environment plant cell nuclei. Here, we analyzed sequence, composition epigenetic makeup peculiar non‐pericentromeric segments Australian crucifer Ballantinia antipoda . By combination high...

Modified gravitational wave propagation is a smoking gun of modifications gravity at cosmological scales, and can be the most promising observable for testing such theories. The observation waves (GW) in recent years has allowed us to start probing this effect, here we briefly review two ways it. We will show that, already with current network detectors, it possible reach an interesting accuracy estimation $\Xi_0$ parameter (that characterizes modified propagation, $\Xi_{0, {\rm GR}} = 1$)...

In plants, a particular class of small non-coding RNAs, short interfering can serve as signal to induce cytosine methylation at homologous genomic DNA regions in the nucleus. If targeted have promoter function, this RNA-directed (RdDM) result transcriptional gene silencing (TGS). transgenes provides versatile system for study epigenetic regulation plants. our experimental setup Arabidopsis thaliana, transcription promoter-inverted repeat RNA that triggers de novo and TGS nopaline synthase...

Modified gravitational wave (GW) propagation is a generic phenomenon in modified gravity. It affects the reconstruction of redshift coalescing binaries from luminosity distance measured by GW detectors, and therefore actual masses component compact stars observed (`detector-frame') masses. We show that, thanks to narrowness mass distribution binary neutron stars, this effect can provide clear signature gravity, particularly for redshifts explored third generation detectors such as Einstein...