Male-Specific Association between a γ-Secretase Polymorphism and Premature Coronary Atherosclerosis

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Vascular damage in testicular cancer patients: A study on endothelial activation by bleomycin and cisplatin in vitro

Following treatment with bleomycin- and cisplatin-containing chemotherapy, testicular cancer patients frequently develop vascular complications, which may result from damage to endothelial cells. Understanding bleomycin- and cisplatin-induced endothelial alterations may help to develop strategies to prevent or reduce vascular toxicity. The effects of bleomycin and cisplatin on proliferation and apoptosis of the human dermal microvascular endothelial cell line HMEC-1 were determined. In addition, modulation of drug-induced cytotoxicity by the free radical scavenger amifostine, the low molecular weight heparin dalteparin, the iron-chelator dexrazoxane, the HMG-CoA reductase inhibitor rosuvastatin and the PPAR agonist troglitazone was tested. Furthermore, the effects of bleomycin and cisplatin oil endothelial activation measured by the expression of the intercellular adhesion rnolecule-1 (ICAM-1) and on two main proteins involved in fibrinolysis, tissue-type plasminogen activator (tPA) and plasminogen activator inhibitor type I (PAI-1), were measured. Decreased endothelial cell survival induced by bleomycin and cisplatin coincided with the induction of apoptosis. Only troglitazone was able to protect the endothelial cells from both bleomycin- and cisplatin-induced cytotoxicity. At high concentrations, amifostine and dexrazoxane also protected HMEC-1 from drug-induced cytotoxicity. However, due to the required high (toxic) concentrations of both modulators no absolute cell survival benefit could be achieved. Both bleomycin and cisplatin induced up-regulation of ICAM-1, tPA and PAI-1. Summarizing, bleomycin and cisplatin induce alterations in the function of endothelial cells regarding proliferation, inflammation and fibrinolysis in vitro. Strategies aimed at these functions should be developed in order to ameliorate or prevent cytostatic agent-induced vascular damage

Genotype and allele frequencies for the <i>APH1B</i> Phe217Leu variation in a Dutch case-control study on premature coronary atherosclerosis.

<p>Standard Chi-square tests were applied to evaluate the association with premature coronary atherosclerosis. OR = odds ratio; *<i>p</i><0.05.</p

γ-Secretase cleavage activity of human wild-type APH1B (Phe217) and human mutant APH1B (Leu217) stably transfected into <i>Aph1abc^−/−</i> mouse embryonic fibroblast cells.

<p>The levels of the C-terminal fragments (CTFs) of N-cadherin, APP and syndecan 3 were analysed in cells stably transfected with human APH1B Phe217 (WT) or human APH1B Leu217 (MUT). In the MUT cells, the CTF levels of N-cadherin and APP were slightly increased (1.1-fold and 1.2-fold, respectively), but the difference failed to reach significance. The levels of syndecan-3-CTF were significantly increased (1.6-fold; p = 0.044), indicating a reduced γ-secretase cleavage activity. Bars represent quantifications of CTF signals normalized to full-length protein levels in eight stably transfected cell lines with the average level in WT cell lines set to 100%. *: <i>p</i><0.05.</p

Clinical and biochemical characteristics of the premature coronary atherosclerosis patients with and without the <i>APH1B</i> 217Leu allele.

<p>Values are given as mean levels±SD or as percentages. N, number of individuals tested; BMI, body mass index; LDL, low density lipoprotein; HDL, high density lipoprotein; Apo, apolipoprotein. *<i>p</i><0.05.</p

Alignment of vertebrate and invertebrate amino acid sequences of the region within APH1 surrounding residue 217.

<p>A black background indicates identical amino acid residues; a grey background indicates a conservative amino acid change. Abbreviations: Hs, <i>Homo sapiens</i>; Pt, <i>Pan troglodytes</i>; Rn, <i>Rattus norvegicus</i>; Mm, <i>Mus musculus</i>; Xt, <i>Xenopus tropicalis</i>; Dr, <i>Danio rerio</i>; Dm, <i>Drosophila melanogaster</i>; Ce, <i>Caenorhabditis elegans</i>; At, <i>Arabidopsis thaliana</i>. Sequences were aligned using Vector NTI (9.0) with default parameters.</p

Future of Dutch NGS-Based Newborn Screening:Exploring the Technical Possibilities and Assessment of a Variant Classification Strategy

In this study, we compare next-generation sequencing (NGS) approaches (targeted panel (tNGS), whole exome sequencing (WES), and whole genome sequencing (WGS)) for application in newborn screening (NBS). DNA was extracted from dried blood spots (DBS) from 50 patients with genetically confirmed inherited metabolic disorders (IMDs) and 50 control samples. One hundred IMD-related genes were analyzed. Two data-filtering strategies were applied: one to detect only (likely) pathogenic ((L)P) variants, and one to detect (L)P variants in combination with variants of unknown significance (VUS). The variants were filtered and interpreted, defining true/false positives (TP/FP) and true/false negatives (TN/FN). The variant filtering strategies were assessed in a background cohort (BC) of 4833 individuals. Reliable results were obtained within 5 days. TP results (47 patient samples) for tNGS, WES, and WGS results were 33, 31, and 30, respectively, using the (L)P filtering, and 40, 40, and 38, respectively, when including VUS. FN results were 11, 13, and 14, respectively, excluding VUS, and 4, 4, and 6, when including VUS. The remaining FN were mainly samples with a homozygous VUS. All controls were TN. Three BC individuals showed a homozygous (L)P variant, all related to a variable, mild phenotype. The use of NGS-based workflows in NBS seems promising, although more knowledge of data handling, automated variant interpretation, and costs is needed before implementation.</p