Influence of Uraemia on the Determination of Blood Glycohaemoglobin by HPLC, Electrophoresis and Affinity Chromatography in Diabetic and Non-Diabetic Patients

Peer Reviewe

Mild temperature shock affects transcription of yeast ribosomal protein genes as well as the stability of their mRNAs.

Shifting the temperature of a yeast culture from 23 degrees to 36 degrees C results in a sudden and severe (greater than 85%) decline in the cellular levels of ribosomal protein (rp-)mRNAs. Recovery during continued growth at 36 degrees C occurs within 1 h. The use of hybrid genes carrying different portions of the region upstream of the gene coding for ribosomal protein L25 revealed that this characteristic, coordinate temperature shock phenomenon does not depend on the presence of specific upstream DNA sequences. Analysis of a heterologous gene carrying a synthetic UASrpg (upstream activation site of yeast ribosomal protein genes) provided conclusive evidence that the rp-characteristic, transient heat shock response is not mediated through the UASrpg elements. The addition of the transcription inhibitor 1,10-phenantroline prior to a 23 degrees to 36 degrees C heat shock inhibited the severe decline of the rp-mRNA levels. The latter observation indicates that transcription is required for the rp-gene- specific response to heat shock. A milder temperature shift, from 23 degrees to 30 degrees C, gave rise to a two-fold decrease in mRNA levels for all genes studied, both ribosomal and non-ribosomal. Together, these results indicate that a temperature shift causes a temporary general transcriptional arrest in yeast cells, resulting in an over-all decrease in mRNA levels. In addition, an enhanced nucleolytic break-down of pre-existing rp-mRNAs accounts for the dramatic drop in the steady state amounts of these mRNAs observed upon a 23 degrees----36 degrees C shift. This enhanced breakdown is caused directly or indirectly by a factor whose synthesis is induced by the heat shock treatment

The extended promoter of the gene encoding ribosomal protein S33 in yeast consists of multiple protein binding elements.

At least 4 different, protein binding cis-acting elements are present in the upstream region of the S33-gene. The major protein binding site is situated between positions -148 and -163 relative to the ATG start codon. It binds a trans-acting factor designated SUF (S33 Upstream Factor). When yeast cells are growing on glucose, deletion of this site results in a decrease of transcription of 50%. Using ethanol as a carbon-source, deletion of the SUF-responsive site lowers the transcription as much as 80%. A second protein binding site is found between positions -85 and -105. Only extracts from glucose-grown cells contain a factor that is able to bind to this site in vitro. A third protein binding site was found using a protein extract from ethanol-grown cells. This site, which is located quite close to the transcriptional start site, is probably responsible for the 20% residual transcription when the SUF binding site is removed. Finally, a site far upstream was found, which binds a protein from both glucose-grown and ethanol-grown cells. This site may function as an upstream repression site which is only functional when a non-fermentable carbon-source is used. Taking these findings into account, we present a model for the carbon-source dependent transcription activation of the gene encoding S33

Transcriptional control of yeast ribosomal protein synthesis during carbon-source upshift.

Shifting a yeast culture from an ethanol-based medium to a glucose-based medium causes a coordinate increase of the cellular levels of ribosomal protein mRNAs by about a factor 4 within 30 min. Making use of hybrid genes encompassing different portions of the 5'-flanking region of the L25-gene, we could show that the increase in mRNAs is a transcriptional event, mediated through DNA sequences upstream of the ribosomal protein (rp) genes. Further analysis revealed that sequence elements are involved that many rp-genes have in common and that previously were identified as transcription activation sites (RPG-boxes or UASrpg). Using appropriate deletion mutants of the fusion genes we could demonstrate that a single RPG-box is sufficient for the transcriptional upshift. In addition, both copy genes encoding rp28 which differ considerably in their extent of transcriptional activity, show the upshift effect in a proportional manner. Definite proof for the role of the UASrpg in nutritional regulation was obtained by examining the effect of a synthetic RPG-box on transcription

Objectives and Design of BLEEDS: A Cohort Study to Identify New Risk Factors and Predictors for Major Bleeding during Treatment with Vitamin K Antagonists

<div><p>Background</p><p>Risk scores for patients who are at high risk for major bleeding complications during treatment with vitamin K antagonists (VKAs) do not perform that well. BLEEDS was initiated to search for new biomarkers that predict bleeding in these patients.</p><p>Objectives</p><p>To describe the outline and objectives of BLEEDS and to examine whether the study population is generalizable to other VKA treated populations.</p><p>Methods</p><p>A cohort was created consisting of all patients starting VKA treatment at three Dutch anticoagulation clinics between January-2012 and July-2014. We stored leftover plasma and DNA following analysis of the INR.</p><p>Results</p><p>Of 16,706 eligible patients, 16,570 (99%) were included in BLEEDS and plasma was stored from 13,779 patients (83%). Patients had a mean age of 70 years (SD 14), 8713 were male (53%). The most common VKA indications were atrial fibrillation (10,876 patients, 66%) and venous thrombosis (3920 patients, 24%). 326 Major bleeds occurred during 17,613 years of follow-up (incidence rate 1.85/100 person years, 95%CI 1.66–2.06). The risk for major bleeding was highest in the initial three months of VKA treatment and increased when the international normalized ratio increased. These results and characteristics are in concordance with results from other VKA treated populations.</p><p>Conclusion</p><p>BLEEDS is generalizable to other VKA treated populations and will permit innovative and unbiased research of biomarkers that may predict major bleeding during VKA treatment.</p></div

Incidence rates of bleeding events stratified by time since start of VKA treatment.

<p>Incidence rates of bleeding events stratified by time since start of VKA treatment.</p

Time in therapeutic range per day after starting VKA treatment.

<p>Time in therapeutic range per day after starting VKA treatment.</p

Number of bleeding events stratified by area.

<p>Number of bleeding events stratified by area.</p