Einfluss von Inflammation und Diabetes auf die Pathophysiologie des duktalen Adenokarzinoms des Pankreas

Verglichen mit den normoglykämischen Kontrolltieren entwickelten die diabetischen Tiere Karzinome mit einem signifikant höheren Tumorgewicht. Dabei korrelierte das höhere Tumorgewicht der diabetischen Tiere mit einer erhöhten Proliferation der Karzinomzellen, während der Diabetes nur einen geringen Einfluss auf die Apoptose und Nekrose in den Karzinomen hatte. Die chronische Pankreatitis hatte keinen wesentlichen Einfluss auf die Tumorgröße, die Proliferation oder den Zelltod, führte jedoch zu einer gesteigerten Lipaseaktivität im Plasma und zu einer Atrophie des Pankreasparenchyms

Einfluss von Inflammation und Diabetes auf die Pathophysiologie des duktalen Adenokarzinoms des Pankreas

Verglichen mit den normoglykämischen Kontrolltieren entwickelten die diabetischen Tiere Karzinome mit einem signifikant höheren Tumorgewicht. Dabei korrelierte das höhere Tumorgewicht der diabetischen Tiere mit einer erhöhten Proliferation der Karzinomzellen, während der Diabetes nur einen geringen Einfluss auf die Apoptose und Nekrose in den Karzinomen hatte. Die chronische Pankreatitis hatte keinen wesentlichen Einfluss auf die Tumorgröße, die Proliferation oder den Zelltod, führte jedoch zu einer gesteigerten Lipaseaktivität im Plasma und zu einer Atrophie des Pankreasparenchyms

Selection-Independent Generation of Gene Knockout Mouse Embryonic Stem Cells Using Zinc-Finger Nucleases

Gene knockout in murine embryonic stem cells (ESCs) has been an invaluable tool to study gene function in vitro or to generate animal models with altered phenotypes. Gene targeting using standard techniques, however, is rather inefficient and typically does not exceed frequencies of 10−6. In consequence, the usage of complex positive/negative selection strategies to isolate targeted clones has been necessary. Here, we present a rapid single-step approach to generate a gene knockout in mouse ESCs using engineered zinc-finger nucleases (ZFNs). Upon transient expression of ZFNs, the target gene is cleaved by the designer nucleases and then repaired by non-homologous end-joining, an error-prone DNA repair process that introduces insertions/deletions at the break site and therefore leads to functional null mutations. To explore and quantify the potential of ZFNs to generate a gene knockout in pluripotent stem cells, we generated a mouse ESC line containing an X-chromosomally integrated EGFP marker gene. Applying optimized conditions, the EGFP locus was disrupted in up to 8% of ESCs after transfection of the ZFN expression vectors, thus obviating the need of selection markers to identify targeted cells, which may impede or complicate downstream applications. Both activity and ZFN-associated cytotoxicity was dependent on vector dose and the architecture of the nuclease domain. Importantly, teratoma formation assays of selected ESC clones confirmed that ZFN-treated ESCs maintained pluripotency. In conclusion, the described ZFN-based approach represents a fast strategy for generating gene knockouts in ESCs in a selection-independent fashion that should be easily transferrable to other pluripotent stem cells

Heart failure after pressure overload in autosomal-dominant desminopathies: Lessons from heterozygous DES-p.R349P knock-in mice

Background Mutations in the human desmin gene (DES) cause autosomal-dominant and -recessive cardiomyopathies, leading to heart failure, arrhythmias, and AV blocks. We analyzed the effects of vascular pressure overload in a patient-mimicking p.R349P desmin knock-in mouse model that harbors the orthologue of the frequent human DES missense mutation p. R350P. Methods and results Transverse aortic constriction (TAC) was performed on heterozygous (HET) DES-p.R349P mice and wild-type (WT) littermates. Echocardiography demonstrated reduced left ventricular ejection fraction in HET-TAC (WT-sham: 69.5 ± 2.9%, HET-sham: 64.5 ± 4.7%, WTTAC: 63.5 ± 4.9%, HET-TAC: 55.7 ± 5.4%; p<0.01). Cardiac output was significantly reduced in HET-TAC (WT sham: 13088 ± 2385 μl/min, HET sham: 10391 ± 1349μl/min, WT-TAC: 8097 ± 1903μl/min, HET-TAC: 5793 ± 2517μl/min; p<0.01). Incidence and duration of AV blocks as well as the probability to induce ventricular tachycardias was highest in HET-TAC. We observed reduced mtDNA copy numbers in HET-TAC (WT-sham: 12546 ± 406, HET-sham: 13526 ± 781, WT-TAC: 11155 ± 3315, HET-TAC: 8649 ± 1582; p = 0.025), but no mtDNA deletions. The activity of respiratory chain complexes I and IV showed the greatest reductions in HET-TAC. Conclusion Pressure overload in HET mice aggravated the clinical phenotype of cardiomyopathy and resulted in mitochondrial dysfunction. Preventive avoidance of pressure overload/arterial hypertension in desminopathy patients might represent a crucial therapeutic measure

Rapid and automated risk stratification by determination of the aortic stiffness in healthy subjects and subjects with cardiovascular disease.

BackgroundAortic stiffness is an independent predictor of cardiovascular morbidity and mortality; thus, simple, rapid and preferably automated techniques are indispensable for pursuing a global risk stratification approach. We present an oscillometric technique for determination of the carotid-femoral pulse wave velocity (cfPWV), including the diagnostic accuracy, sensitivity and specificity, with emphasis on the training curve and procedural duration.MethodsIn a single-centre crossover study, we evaluated subjects free of known cardiovascular disease (CVD), subjects with CVD and a subgroup of subjects with peripheral artery disease (PAD) in terms of ankle-brachial index (ABI) and PWV measurements determined by oscillometry compared to tonometry. Pearson's correlation analysis was used to assess the relationship of the PWV measurements determined by both methods. Moreover, the time and cost of the examinations were compared.ResultsA total of 176 study subjects underwent assessments to obtain oscillometric and tonometric PWV measurements. The CVD-free subjects (n = 59) were younger (60.4±15.6 vs. 67.5±12.9 years, p = 0.003) than the subjects with CVD (n = 117). The PWV measurements showed significant correlations in CVD-free subjects (r = 0.797, pConclusionUsing a simple and rapid automated oscillometric method, we achieved good diagnostic accuracy for the determination of aortic stiffness through the PWV in both subjects with and without CVD. This method might be helpful in daily practice in terms of saving time and reducing procedural complexity for screening for cardiovascular morbidities and vascular damage in cases of atherosclerosis

Brief report - Telomere length is a poor biomarker to predict 1-year mortality or cardiovascular comorbidity in patients with transcatheter aortic valve replacement.

BackgroundTranscatheter aortic valve replacement (TAVR) is a therapeutic option for patients with aortic valve stenosis at increased surgical risk. Telomeres are an established marker for cellular senescence and have served to evaluate cardiovascular diseases including severe aortic valve stenosis. In our study, we hypothesized that telomere length may be a predictor for outcome and associated with comorbidities in patients with TAVR.Methods and resultsWe analyzed leucocyte telomere length from 155 patients who underwent TAVR and correlated the results with 1-year mortality and severe comorbidities. The cohort was subdivided into 3 groups according to telomere length. Although a trend for a positive correlation of telomere length with a lower EuroSCORE could be found, telomere length was not associated with survival, aortic valve opening area or cardiovascular comorbidities (peripheral, coronary or cerebrovascular disease). Interestingly, long telomeres were significantly correlated to a reduced left ventricular ejection fraction (LVEF).ConclusionIn elderly patients with severe aortic valve stenosis, leucocyte telomere length did not predict post-procedural survival. The correlation between long telomere length and reduced LVEF in these patients deserves further attention

Brief report – Telomere length is a poor biomarker to predict 1-year mortality or cardiovascular comorbidity in patients with transcatheter aortic valve replacement

Background: Transcatheter aortic valve replacement (TAVR) is a therapeutic option for patients with aortic valve stenosis at increased surgical risk. Telomeres are an established marker for cellular senescence and have served to evaluate cardiovascular diseases including severe aortic valve stenosis. In our study, we hypothesized that telomere length may be a predictor for outcome and associated with comorbidities in patients with TAVR. Methods and results: We analyzed leucocyte telomere length from 155 patients who underwent TAVR and correlated the results with 1-year mortality and severe comorbidities. The cohort was subdivided into 3 groups according to telomere length. Although a trend for a positive correlation of telomere length with a lower EuroSCORE could be found, telomere length was not associated with survival, aortic valve opening area or cardiovascular comorbidities (peripheral, coronary or cerebrovascular disease). Interestingly, long telomeres were significantly correlated to a reduced left ventricular ejection fraction (LVEF). Conclusion: In elderly patients with severe aortic valve stenosis, leucocyte telomere length did not predict post-procedural survival. The correlation between long telomere length and reduced LVEF in these patients deserves further attention

MOESM1 of Analysis of Axin2 expression and function in murine models for pancreatic cancer

Additional file 1. In the Supplemental Material Section results from beta-galactosidase stainings, blood glucose concentrations, body weights, tumor weights, as well as number of leucocytes, lymphocytes and monocytes plus ganulocytes are presented

Assessment of metaphase chromosomes and pluripotency.

<p>(<b>A–C</b>) Chromosome analysis. Metaphase spreads of BK4-G3.16 cells were stained with Giemsa. A representative picture of ∼20 analyzed metaphase spreads per clone is shown. (<b>D–L</b>) Assessment of pluripotency. ZFN-treated BK4-G3.16 cells were injected subcutaneously into immunodeficient mice and teratomas removed after 4–8 weeks. Histological analysis using hematoxylin/eosin staining revealed tissues derived from ectoderm (D, G, J), mesoderm (E, H, K) and endoderm (F, I, L). Scale bars = 100 µm for (D–G) and 50 µm for (H–L).</p