Objective: To assess feasibility and accuracy of oscillometric cardiac output (CO) calculation. Design and method: A prospective comparison of oscillometric CO determinations and reference themodilution CO measurements was performed in 38 intensive care unit patients (June 2015 to June 2016). Thermodilution CO was obtained using transpulmonary technique (PiCCO®, Pulsion Medical Systems, Feldkirchen, Germany), and oscillometric CO was calculated using the Tel-O-GRAPH® (TG) non-invasive blood pressure measurement and hemodynamic assessment device (IEM, Stolberg, Germany). CO was indexed to body surface area and is referred to as cardiac index (CI). Bland Altman analysis was employed to assess differences between oscillometric and reference CI as well as non-invasive (TG) and invasive (PiCCO®) blood pressure (BP) measurements. Results: Two-thirds of the study population was male (68.4%), mean age was 68 years. Almost all patients were mechanically ventilated (95%) at study entry, and vasopressor therapy was required in 76%. Oscillometric CI determination yielded significantly lower results than the reference method (3.8 ± 1.22 l/min*m2 vs. 2.73 ± 0.50 l/min*m2, p < 0.0001) with a bias of 1.08 l/min*m2 and limits of agreement of ± 2.23 l/min*m2 (percentage error 62%). With TG, both measured brachial (117.5 ± 16.0 mmHg) and estimated aortic (107.2 ± 15.6) systolic BP were significantly lower than invasive PiCCO® femoral BP (123.6 ± 17.4 mmHg, p = 0.005 and p < 0.0001) with a bias of 5.9 mmHg and 16.1 mmHg, respectively, and limits of agreement of ± 23.46 mmHg and ± 23.79 mmHg.Conclusions: TG significantly underestimates CO in hemodynamically unstable patients. This finding could in part be explained by the significantly lower BP readings obtained with TG. In addition, Bland-Altman analysis revealed a linear relationship between CO values and divergence of TG CO from reference measurement (figure). A correction factor could potentially alleviate CO underestimation with TG, and thus make this noninvasive easy method of CO determination more reliable

Objective: Cardiovascular disease are the leading cause of death. Changes within the vascular wall as mineralization of vascular smooth muscle cells is one pathogenesis of cardiovascular alterations. Increased oxidative stress and cellular senescence of vascular cells are main factors in the pathogenesis. The cytostatic drug doxorubicin (DOX) induces the production of reactive oxygen species (ROS), activates cell apoptosis mechanisms and promote cellular senescence. The aim of this study is to investigate the effect of DOX on vascular smooth muscle cell mineralization. Design and method: Vascular smooth muscle cells of rats (rVSMC) were used for in vitro experiments and aortic rings of rats for ex vivo experiments. Calcification of cells was induced using a high phosphate medium. Calcium content was quantified photometrical via o-cresol-phthalain method. Gene expression of p53, p21, alkaline phosphate (ALP), osteopontin (OPN) and core binding factor-α (cbfa1) was measured via real-time PCR. ALP activity was quantified using the p-nitro-phenol assay. Results: Gene expression of senescence markers as p21 and p53 dose-dependently increase upon DOX stimulation for 24 h, 48 h and 72 h. Gene expression of ALP, OPN and cbfa1 also increase dose-dependently upon DOX stimulation for 24 to 72 h. ALP enzyme activity is significantly induced by DOX stimulation during 14 days of stimulation. Long-term treatment of rVSMC with DOX for 14 days significantly increased the mineralization of the cells as quantified by the extracellular calcium content. Ex vivo experiments with aortic rings from rats confirmed the findings. Here, DOX stimulation for 14 days induces mineralization that could be found located within the media of the vessel wall by histological staining with Alizarin Red and van Kossa. Conclusions: DOX is a robust inductor of cellular senescence and mineralization of smooth muscle cells. The data let suggest that side effects of cytostatic treatment with DOX may contribute to the high cardiovascular risk of patients by vascular aging and stiffening.

Objective: Central blood pressure (CBP) measurement might precise the individual cardiovascular risk of the patient. We sought to evaluate the accuracy of the CBP calculation with a new oscillometric device Tel-O-GRAPH® (I.E.M., Stolberg, Germany) in different common clinical situations.Design and method: 103 subjects were prospectively included in the study. The performance accuracy of Tel-O-GRAPH® was assessed in comparison to Sphygmocor® (AtCor Medical, West Ryde, NSW, Australia) using Bland-Altman approach. The robustness of the values with Tel-O-GRAPH® was evaluated in supine and seated positions as well as for experienced and inexperienced user. Results: Mean age of the study population was 60 ± 17.9 years. 56.6% were male and mean BMI was 26.5 ± 4.9 kg/sqm. The mean systolic CBP was 126.7 ± 21.9 mmHg measured with Tel-O-GRAPH® and 126.6 ± 22.5 mmHg measured with Sphygmocor®. Good agreement between Tel-O-GRAPH® and Sphygmocor® for CBP could be shown (mean -0.3 ± 6.7 mmHg; Person's r = 0.95; p < 0.0001). The mean difference of CBP with Tel-O-GRAPH® was 1.5 ± 6.8 mmHg (r = 0.9; p < 0.0001) and -1.4 ± 5.0 (r = 0.97; p < 0.0001) mmHg between supine vs. seated position and between experienced vs. inexperienced user respectively. Conclusions: Tel-O-GRAPH® calculates CBP easy and quickly using “one button press” procedure. We observed a high CBP measurement accuracy compared to Sphygmocor®. Given stable brachial blood pressure measured CBP values seem to remain robust independently of body position or operator experience.

Objective: It is a well-known fact that cardiovascular alterations like arteriosclerosis lead to a high cardiovascular morbidity and mortality. The cellular and molecular mechanisms of the mineralization process are delved for by utilizing different experimental settings, e.g., in vitro models with vascular smooth muscle cells, ex vivo models using aortic rings as well as in vivo rat or mice models. The aim of this study was to establish a novel model for vascular calcification studies via ex vivo perfusion of a thoracic aortic tissue. Design and method: The design and conception of the perfusion chamber were done pursuant to the requirements. The thoracic aorta of a Wistar rat was dissected and all aortic branches were coagulated. The aorta was perfused for 14 days at 37°C in a humified atmosphere. Vessel mineralization was detected by quantification of the calcium content (normalized to dry weight) and histological staining (Alizarin Red, von Kossa). Results: The experimental protocol for tissue preparation, buffer and perfusion condition were established. Tissue integrity during perfusion procedure seems to be intact; histo-morphological markers for elastic fibril degradation or necrotic markers are missing. Buffer and perfusion media were tested for induction of non-specific effects. Buffer and control media (w/o perfusion) did not induce vessel mineralization. To induce mineralization a medium containing high phosphate level (known from established ex vivo models) was used. Perfusion of the aortic tissue with this high phosphate medium induced a significant increase of calcium content in the aortic wall compared to perfusion condition with control medium (1.4 ± 0.4 vs. 6.5 ± 1.6 μg/mg; p < 0.05). Histological staining revealed medial located mineralization within the vessel wall. Conclusions: The novel experimental setting provides a novel ex vivo model with solely luminal perfusion, which allows administration of substances in a more physiological manner than well established ex vivo settings for calcification studies. Furthermore, the benefit of easier handling than in vivo models and the advantage of working with tissue make usage for studying signaling pathways and for screening of pharmaceuticals possible.

Objective: There is emerging evidence pointing out the arterial stiffness (AS) as an important independent predictor of cardiovascular events. Carotid-femoral pulse wave velocity (cfPWV) is a best-established surrogate parameter for assessment of AS. Measurement of cfPWV is considered to be gold standard for evaluating AS. Though it has been widely used in clinical and observational studies, its utilisation in the everyday clinical practice is scarce due to technical challenges and procedural pitfalls of the available non-invasive assessment techniques. Emerging oscillometric techniques seem to simplify assessment procedure. We thus aimed to evaluate the accuracy of performance and reproducibility with a new oscillometric device Tel-O-GRAPH® for measurement of PWV. Design and method: 89 patients were prospectively included in the study. Oscillometric calculation of cfPWV was performed with Tel-O-GRAPH® (I.E.M., Stolberg, Germany) and tonometric calculation with Sphygmocor® (AtCor Medical, West Ryde, NSW, Australia). The accuracy assessment was performed according to the requirements of ARTERY Society. Coefficients of variation and intraclass correlation coefficients were used to evaluate the reproducibility. Results: Mean study population age was 48.8 ± 19.1 years. 59.6% of the patients were male and 15.1% were smoker. The mean PWV measured with Tel-O-GRAPH® was 7.8 ± 2.3 m/s, the mean PWV measured with Sphygmocor® was 7.3 ± 1.7 m/s. The mean difference of PWV between devices was 0.49 ± 1.26 m/s (p < 0.0001) and Pearson correlation index was 0.86 (p < 0.0001). The coefficient of variation and intraclass correlation coefficients between single measured PWV values with Tel-O-GRAPH® and Sphygmocor® were 2.38 ± 6.13% vs. 6.3 ± 4.33% (p < 0.05) and 0.99;0.99;0.99 vs. 0,78;0.84;0.71 respectively. Conclusions: We observed that oscillometric calculation of cfPWV with Tel-O-GRAPH® can be done quickly and easy, with very high reproducibility of the values and adequate measurement accuracy compared to Sphygmocor®. Reported data suggest that oscillometry might evolve to a favoured method for assessment of the PWV in the everyday clinical practice and clinical studies due to its easiness of performance with concurrently remained reliability and accuracy.

Endothelial nitric oxide synthase (eNOS) plays a crucial role in vascular homeostasis. Lysophospholipid interaction with sphingosine 1-phosphat (S1P) receptors results in eNOS activation in different cells. In endothelial cells, eNOS activation via S1P1 or S1P3 was shown controversially. The aim of this study is to investigate the meaning of both S1P receptors for eNOS activation in human endothelial cells. Therefore, several S1P1 and S1P3 agonists in combination with antagonists and specific RNAi approach were used. eNOS activation was measured in human umbilical vein endothelial cells (HUVEC) via DAF2-DA-based fluorescence microscopy. For investigation of the signaling pathway, agonists/antagonist studies, RNAi approach, Luminex™ multiplex, and Western Blot were used. In HUVEC, both the S1P1 agonist AUY954 as well as the S1P1,3 agonist FTY720P induced eNOS activation in a time- and dose-dependent manner. Other S1P1 agonists activated eNOS to a lesser extent. The AUY954-induced eNOS activation was blocked by the S1P1 antagonist W146, the combination of W146 and the S1P3 antagonist CAY10444 and the S1P1,3 antagonist VPC23019, but not by CAY10444 indicating the meaning of S1P1 for the AUY954-induced eNOS activation. The FTY720P-induced eNOS activation was blocked only by the combination of W146 and CAY10444 and the combined S1P1,3 antagonist VPC23019, but not by W146 or CAY10444 indicating the importance of both S1P1 and S1P3 for FTY720-induced eNOS activation. These results were confirmed using specific siRNA against S1P1 and S1P3. The S1P1,3 activation results in Akt phosphorylation and subsequent activation of eNOS via phosphorylation at serine1177 and dephosphorylation at threonine495. Beside former investigations with rather unspecific S1P receptor activation these data show potent selective S1P1 activation by using AUY954 and with selective S1P receptor inhibition evidence was provided that both S1P1 and S1P3 lead to downstream activation of eNOS in HUVEC in the same experimental setting. Inhibition or knockdown of one of these receptor subtypes did not abolish the eNOS activation and subsequent NO production.

Abstract Serum amyloid A (SAA) is an apolipoprotein transported within the high density lipoprotein (HDL) in plasma. The SAA plasma levels increase during inflammatory conditions, e.g., in patients with chronic renal failure (CRF). The SAA-dependent reduction in anti-inflammatory condition of HDL and its pro-inflammatory response in smooth muscle cells was shown previously. The aim of this study was to investigate the signaling pathways of SAA in macrophages and therefore its influence on inflammatory vascular disease. THP-1 (human) monocytes were activated via PMA to macrophages and used for monocyte chemoattractant protein-1 (MCP-1) experiments. Murine RAW264.7 monocytes/macrophages were used for nitrite experiments. MCP-1 production was detected by Luminex™ technology. Nitrite production were measured via Griess Assay. Cell viability was determined via MTS assay. SAA accumulates in plasma of patients during conditions of CRF. Whereas there is only a slight increase within CRF stage 1 and 2, the plasma level of SAA further increase during CRF stage 3 to 5. Beside the pro-inflammatory potential of SAA to induce MCP-1 in vascular smooth muscle cells, it also induces MCP-1 secretion in human THP-1 macrophages in a dose-dependent manner. In addition, the production of nitrite was dose-dependently increased in murine RAW264.7 cells. Both, MCP-1 and nitrite production induced by SAA were regulated via TLR and SR-BI receptor activation. The TLR2/4 antagonist oxPAPC and the SR-BI antagonist BLT-1 diminished the SAA-induced MCP-1 and nitrite production. The activation of FPR2 seems not to be involved in the signaling pathway in macrophages after SAA stimulation in that experimental condition. Stimulation with receptor agonists confirmed these findings. The concentration used for agonists/antagonists had no significant influence on cell viability of macrophages. In conclusion, the pro-inflammatory reaction of SAA in macrophages in vitro depends on TLR2/4 and SR-BI activation. The accumulation of SAA plasma levels during CRF may substantially contribute to the increased cardiovascular risk of these patients.

Abstract Purpose: To analyze procedural details, complications and radiation exposure in renal denervation (RDN) using the Medtronic Symplicity® device in the treatment of refractory hypertension. Materials and Methods: Fifty three consecutive patients underwent RDN. The number of ablations per artery, peri-procedural complications, procedure time (PT), fluoroscopy time (FT), dose-area product (DAP) and procedure-related complications were documented. Additionally, the radiation dose was compared between obese (body mass index ≥ 30 kg/m²) and non-obese patients. Results: Bilateral RDN was performed in 50/53 (94 %) cases and with a minimum of 4 ablations per artery in 33/50 (66 %), the mean count being 5.4 (range R: 2 – 13) on the right and 4.3 (R: 1 – 10) on the left. The FT and DAP decreased significantly over the first 12 procedures, reaching a steady state with a median FT of 11.2 min (R: 7.5 – 27) and a median DAP of 4796 cGy × cm² (R: 1076 – 21 371), resulting in an effective dose of 15.7 mSv. The median PT was 57 min (R: 40 – 70). Obese patients had a 3.3-fold higher radiation dose (p < 0.001). We observed one severe spasm and one imminent respiratory depression, both resolved without sequelae. Conclusion: For an experienced interventionalist, RDN has a short learning curve with a low risk profile. The radiation dose does not exceed that of other renal artery interventions, but is explicitly higher in obese patients, who account for a large portion of patients with refractory hypertension.

Jankowski, V.1; Toelle, M.1; Van Der Giet, M.1; Bader, M.2; Zidek, W.1; Jankowski, J.1 Author Information

Jankowski, V.; Toelle, M.; Van Der Giet, M.; Lehmann, K.; Zidek, W.; Jankowski, J. Author Information