Clinical Hemorheology and Microcirculation - Volume 58, issue 1

Authors: Vayá, Amparo | Giménez, Cristina | Sarnago, Ana | Alba, Amparo | Rubio, Olaya | Hernández-Mijares, Antonio | Cámara, Rosa

Article Type: Research Article

Abstract: BACKGROUND: Subclinical hypothyroidism (SCH) has been suggested to be associated with increased cardiovascular risk by different mechanisms. Several cardiovascular risk factors have been analysed, but yielded controversial results. OBJECTIVES: We aimed to analyse whether there are differences in several cardiovascular risk markers, such as lipids, inflammatory parameters: plasma viscosity (PV), fibrinogen and C reactive protein (CRP); homocysteine (Hcy) and red blood cell distribution width (RDW), when comparing SCH and controls. We also analysed which of these parameters predict SCH risk and constitute independent markers. METHODS: We determined PV in a Fresenius capillary plasma viscosimeter, Hcy by a chemiluminiscent enzyme immunoassay, …and biochemical and haematological parameters by conventional laboratory methods in 58 SCH outpatients and 58 controls matched for age and gender. RESULTS: SCH patients did not show statistical differences for glucose, lipids or leucocytes (p > 0.05). However, patients showed a higher prevalence for use of hypolipidaemic drugs, body mass index (BMI), thyroid stimulating hormone (TSH), PV, CRP, fibrinogen, Hcy and RDW (p < 0.05). RDW correlated with inflammation parameters: PV (r = 0.331, p < 0.05), fibrinogen (r = 0.424, p < 0.05), CRP (r = 0.433, p < 0.01) and leucocytes (r = 0.613, p < 0.01). None of the cardiovascular markers correlated with the TSH levels (p > 0.05) In the unadjusted logistic regression analyses, BMI ≥28 kg/m2 , RDW ≥14%, Hcy ≥12 μm/L, fibrinogen ≥400 mg/dL and MCV ≤88 fL increased SCH risk, but only RDW ≥14% and fibrinogen ≥400 mg/dL independently increased this risk in the adjusted logistic regression analyses (OR = 4.68, 95% CI 1.20–18.30 P = 0.026; OR = 3.48, 95% CI 1.08–11.23 P = 0.037). CONCLUSION: SCH patients show a higher cardiovascular risk, characterised by increased PV, fibrinogen, Hcy and RDW. However, only fibrinogen ≥400 mg/dL and RDW ≥14% are independent predictors of SCH. Show more

Keywords: Subclinical hypothyroidism, red blood cell distribution width, homocysteine, fibrinogen, plasma viscosity, cardiovascular risk

DOI: 10.3233/CH-141871

Citation: Clinical Hemorheology and Microcirculation, vol. 58, no. 1, pp. 1-7, 2014

Authors: Klein, S.M. | Prantl, L. | Geis, S. | Eisenmann-Klein, M. | Dolderer, J. | Felthaus, O. | Loibl, M. | Heine, N.

Article Type: Research Article

Abstract: BACKGROUND: Grafting of autologous lipoaspirate for various clinical applications has become a common procedure in clinical practice. With an estimated mortality rate of 10–15 percent, fat embolism is among the most severe complications to be expected after lipofilling therapies. OBJECTIVE: The aim of this study was to determine the level of interstitial pressure after the injection of defined volumes of lipoaspirate into the subcutaneous tissue of female breasts. It was hypothesized, that interstitial pressure levels exceed the physiologic capillary pressure during lipofilling procedures and hence increase the potential risk for fat embolism. Further it was investigated if external tissue expansion …has the potential to significantly reduce interstitial tissue pressure. METHODS: Interstitial pressure was monitored in 36 female patients, that underwent autologous fat injections into the breast. Measurements were conducted with a sensor needle connected to a pressure transducer (LogiCal Pressure Monitoring Kit, Smiths medical int. Ltd., UK). Patients were divided into 4 subcohorts differing in their pre-treatment regimen or local tissue conditions. Pre-treatment consisted of tissue expansion, achieved with the Brava™ (Brava LLC Miami, Fla., USA) vacuum-chamber. RESULTS: The increase in interstitial pressure after injection volumes of 100 ml (p = 0.006), 200 ml (p = 0.000) and between 100 ml and 200 ml (p = 0.004) respectively, were significant in non-mastectomized patients without pre-treatment. Patients pre-treated with Brava™ did not show such statistically significant differences in interstitial pressures before and after the injection of 100 ml and 200 ml of lipoaspirate (p = 0.178). The difference in interstitial pressure in mastectomized patients between 0 ml and 100 ml (p = 0.003), as well as 0 ml and 200 ml (p = 0.028) was significant. The difference in pressures between pre-treated patients and patients without pre-treatment did not differ significantly in the mastectomized patient cohort. CONCLUSION: During lipofilling procedures interstitial pressures are reached that exceed pressure limits defined as hazardous for fat embolism. To date it is unknown what pressure levels need to be considered critical for complications in soft tissue interventions. Further the results indicate higher interstitial pressures for patients that had undergone mastectomy, whereas pre-treatment with external tissue expansion seemed to diminish pressure values. Show more

Keywords: Lipofilling, pressure monitoring, Brava, fat embolism syndrome

DOI: 10.3233/CH-141872

Citation: Clinical Hemorheology and Microcirculation, vol. 58, no. 1, pp. 9-17, 2014

Authors: Wang, Weiwei | Xu, Xun | Li, Zhengdong | Lendlein, Andreas | Ma, Nan

Article Type: Research Article

Abstract: Mesenchymal stem cells (MSCs) are an ideal cell source for tissue engineering and regenerative medicine as they possess self-renewal properties and multilineage differentiation potential. They can be isolated from various tissues and expanded easily through normal cell culture techniques. Genetic modifications of MSCs to further improve their therapeutic efficacy have been widely studied and extensively researched. Compared to viral gene delivery methods, non-viral methods generate less toxicity and immunogenicity and thus represent a promising and effective tool for the genetic engineering of MSCs. In the last decades, various non-viral gene delivery strategies have been developed and some of them have …been applied for MSC transfection. This paper gives an overview of the techniques, influencing factors and potential applications of non-viral methods used for the genetic engineering of MSCs. Show more

Keywords: Mesenchymal stem cells, regenerative medicine, non-viral gene delivery, transfection, lipoplex, polyplex

DOI: 10.3233/CH-141883

Citation: Clinical Hemorheology and Microcirculation, vol. 58, no. 1, pp. 19-48, 2014

Authors: Franke, R.P. | Scharnweber, T. | Fuhrmann, R. | Mrowietz, C. | Wenzel, F. | Krüger, A. | Jung, F.

Article Type: Research Article

Abstract: Different radiographic contrast media (RCM) were shown to induce morphological changes of blood cells (e.g. erythrocytes or thrombocytes) and endothelial cells. The echinocytic shape change of erythrocytes, particularly, affords alterations of the membrane cytoskeleton. The cytoskeleton plays a crucial role for the shape and deformability of the red blood cell. Disruption of the interaction between components of the red blood cell membrane cytoskeleton may cause a loss of structural and functional integrity of the membrane. In this study band4.9 and actin as components of the cytoskeletal junctional complex were examined in human erythrocytes after suspension in autologous plasma or in …plasma RCM mixtures (30% v/v Iodixanol-320 or Iopromide-370) followed by a successive double staining with TRITC-/FITC-coupled monoclonal antibodies. After adding Iopromide-370 to the plasma in practically none of the cells the rounded conformation of the membrane cytoskeleton – as it appeared in cells suspended in autologous plasma – was found. In addition, Iopromide-370 induced thin lines and coarse knob-like structures of band4.9 at the cell periphery while most cell centers were devoid of band4.9, and a box-like arrangement of bands of band4.9. A dissociation between colours red (actin) and green (band4.9) occurred as well. In contrast, erythrocytes suspended in a plasma/Iodixanol-320 mixture showed a membrane cytoskeleton comparable to cells suspended in autologous plasma, Similar results were found with respect to the distribution of actin. This study revealed for the first time RCM-dependent differences in band4.9 activities as possible pathophysiological mechanism for the chemotoxicity of radiographic contrast media. Show more

Keywords: Radiographic contrast media, band4.9 Iodixanol-320, Iopromide-370, actin, chemotoxicity, cytoskeletal junctional complex, erythrocyte

DOI: 10.3233/CH-141894

Citation: Clinical Hemorheology and Microcirculation, vol. 58, no. 1, pp. 49-63, 2014

Authors: Büttner, Petra | Galli, Roberta | Jannasch, Anett | Schnabel, Christian | Waldow, Thomas | Koch, Edmund

Article Type: Research Article

Abstract: Degenerative heart valve disease is a life-threatening disease affecting about 3% of the population over 65 years. Up to date, cardiac surgery with heart valve replacement is the only available therapy. The disease is characterized by degenerative disorganization of the heart valve structure and alterations in the residing cell populations. Causes and mechanisms of disease genesis are still not fully understood and until now pharmacological therapies are not available. Thus there is enormous interest in new technologies that enable a better characterization of structure and composition of diseased valves. Currently most research techniques demand for extensive processing of extracted valve …material. We present a novel approach combining coherent anti-Stokes Raman scattering, endogenous two-photon excited fluorescence and second harmonic generation. Cusp constituents can be examined simultaneously, three-dimensionally and without extensive manipulation of the sample enabling impressive insights into a complex disease. Show more

Keywords: Heart valve disease, aortic valve stenosis, aortic valve insufficiency, fibrosis, coherent anti-Stokes Raman scattering, endogenous two-photon excited fluorescence, second harmonic generation, nonlinear optical microscopy

DOI: 10.3233/CH-141882

Citation: Clinical Hemorheology and Microcirculation, vol. 58, no. 1, pp. 65-75, 2014

Authors: Vahldiek, Janis L. | Lehmann, Kai S. | Poch, Franz | Zurbuchen, Urte | Kreis, Martin E. | Gemeinhardt, Ole | Hamm, B. | Niehues, Stefan M.

Article Type: Research Article

Abstract: Radiofrequency ablation (RFA) has shown to be a reasonable alternative for the treatment of hepatic tumors and metastases although multiple limitations remain. Cooling effects due to larger vessels can prevent complete coverage and may lead to early tumor relapse. This preliminary in vivo pig study combines the use of multipolar RFA with three applicators (six electrodes) and interrupted liver perfusion using Pringle's maneuver to overcome the most serious limitations. Furthermore, immediate detection of incomplete RFA is important to revise ablation. We used contrast enhanced computed tomography (CECT) to evaluate post ablation results in comparison to macroscopic images in healthy pig …liver. We found significantly (p = 0.001) larger ablation zones and no affection by larger vessels with interrupted liver perfusion. This allows effective RFA for larger tumors. Immediate postinterventional CECT provided comparable results (r = 0.985) to macroscopic evaluation. Show more

Keywords: Liver perfusion, animals, catheter ablation, electrodes, imaging, three-dimensional/*methods, multidetector computed tomography, radiography, Interventional/methods, *contrast media

DOI: 10.3233/CH-141886

Citation: Clinical Hemorheology and Microcirculation, vol. 58, no. 1, pp. 77-87, 2014

Authors: Greis, C.

Article Type: Research Article

Abstract: Ultrasound contrast agents have substantially extended the clinical value of ultrasound, allowing the assessment of blood flow and distribution in real-time down to microcapillary level. Selective imaging of contrast agent signals requires a contrast-specific imaging mode on the ultrasound scanner, allowing real-time separation of tissue and contrast agent signals. The creation of a contrast image requires a specific interaction between the insonated ultrasound wave and the contrast agent microbubbles, leading to persistent oscillation of the bubbles. Several technical and procedural parameters have a significant influence on the quality of CEUS images and should be controlled carefully to obtain good image …quality and a reliable diagnosis. Achieving the proper balance between the respective parameters is a matter of technical knowledge and experience. Appropriate training and education should be mandatory for every investigator performing CEUS examinations. Show more

Keywords: Ultrasound, microbubble, contrast, CEUS, mechanical index, gain, frame rate, contrast agent dose, presets, image quality, artefacts

DOI: 10.3233/CH-141873

Citation: Clinical Hemorheology and Microcirculation, vol. 58, no. 1, pp. 89-95, 2014

Authors: Nantais, Jordan | Dumbarton, Tristan C. | Farah, Nizam | Maxan, Alexander | Zhou, Juan | Minor, Samuel | Lehmann, Christian

Article Type: Research Article

Abstract: Methylene blue (MB) has been used with some success as a treatment for the vasoplegia of vasopressor-refractory septic shock. The putative mechanism of action of MB is the inhibition of endothelial nitric oxide within the microvasculature and improved responsiveness to endogenous catecholamines (norepinephrine (NE)). However, to date, no study has demonstrated the microcirculatory effect of methylene blue in septic shock. The objective of this randomized, controlled, animal study was to show, in an experimentally-induced, septic shock model in rats, the effects of MB and NE on global hemodynamics and the microcirculation. Mean arterial pressure (MAP) was drastically reduced following bacterial …endotoxin (lipopolysaccharide, LPS) administration in animals not receiving vasopressors. Only the combination of NE + MB restored MAP to control levels by the end of the three hour experiment. Intravital microscopy of the microcirculation was performed in the terminal ileum in order to examine functional capillary density in intestinal muscle layers and the mucosa, as well as leukocyte activation in venules (rolling, adhesion to the endothelium). Untreated LPS animals showed a significant increase in leukocyte adhesion and a decrease in capillary perfusion in the intestinal microcirculation. In groups receiving NE or NE+MB, we observed a significant decrease in leukocyte adhesion and improved functional capillary density, indicating that microvasculature function was improved. This study suggests that methylene blue may be able to improve hemodynamics while preserving microvascular function in septic shock. Show more

Keywords: Methylene blue, norepinephrine, septic shock, microcirculation, intravital imaging

DOI: 10.3233/CH-141874

Citation: Clinical Hemorheology and Microcirculation, vol. 58, no. 1, pp. 97-105, 2014

Authors: Hiebl, B. | Hopperdietzel, C. | Hünigen, H. | Dietze, K. | Jung, F. | Niehues, S.M.

Article Type: Research Article

Abstract: Implantable long-term central venous port systems (CVPS) are widely used as a permanent means of accessing the vascular system for intravenous delivery of drugs, parenteral nutrition, blood transfusion, and blood sampling. These systems allow easy and repetitive puncture without causing much damage to the vessels. However, the body foreign surface of CVPS induces an inflammatory response with varying intensity (depending on the implant materials) that leads to formation of a fibrous tissue capsule around the implant. This study was designed to investigate the influence of bacterial infection on the tissue reaction induced by implanted CVPS in adult patients. 20 patients …(9 women, 11 men, 58 ± 14 yrs of age) were included in this study. These patients received explantation of a polysulfone based CVPS (ChemoSite™, Covidien, Mansfield, USA) due to port related infections (patients with bacterial infections at the implantation site: group A, 5 men, 1 women) or to other reasons such as termination of treatment, thrombosis, or CVPS dysfunction (patients without bacterial infections, group B, 6 men, 8 women) 299.9 ± 261.2 days after CVPS implantation. A sample of the encapsulating tissue covering the CVPS together with surrounding tissue (at least 1 × 1 cm2 ) was placed in a small container with fixing agent, a buffered neutral 4% formalin solution (pH 7). Histological sections of the samples were prepared for light microscopic analysis after paraffin embedding. Sections of 3 μm were cut and stained with haematoxylin and eosin, Weigert's elastic stain, and Heidenhain's azan stain. There was no difference in thickness, collagen and elastin content, or cell and capillary density of the fibrous capsule between both groups. Due to the wound healing reaction involving angiogenesis and fibroblast activation cell density and number of capillaries in the capsule tissue of all patients showed a positive correlation (r = 0.45, p < 0.05). However, the study demonstrated that at the end of the foreign body reaction the artificial tissue layer which covers the CVPS after implantation due to foreign body reaction shows only low reactivity towards infections. Show more

Keywords: Encapsulation, infection, central venous access system, biomaterial

DOI: 10.3233/CH-141881

Citation: Clinical Hemorheology and Microcirculation, vol. 58, no. 1, pp. 107-113, 2014

Authors: Rjosk-Dendorfer, D. | Reu, S. | Deak, Z. | Hetterich, H. | Kolben, T. | Reiser, M. | Clevert, D.A.

Article Type: Research Article

Abstract: PURPOSE: To evaluate the use of color Doppler sonography and free hand sonoelastography in the assessment of breast fibroadenomas according to their size and shape. MATERIALS AND METHODS: From December 2012 to March 2013 women with 16 solid breast masses, classified as BI-RADS category 3 or 4 were examined with B-mode ultrasound, sonoelastography and color Doppler sonography. Lesions were subdivided according to their shape in round, ovoid or macrolobulated and according to their size (<2.0 cm or ≥2.0 cm). Two readers assessed sonoelastographic findings at 12.5 MHz using the tsukuba elasticity score and results of Doppler sonography using a score …of 0, 1 or 2, depending on the degree of perfusion. RESULTS: Among the 16 examined lesions 3 showed a round shape, 9 were ovoid and in 4 cases a macrolobulated appearance could be seen. No significant differences concerning Tsukuba-score depending on various shapes of fibroadenomas in B-mode sonography could be shown (p = 0.91) and also comparison of Tsukuba-scores and size of masses revealed no significant differences (p = 1.0). Sizes of fibroadenomas ≥2 cm were significantly associated with an increased vascularization of the lesions (p = 0.04) and a macrolobulated appearance in B-mode sonography (p = 0.04). CONCLUSION: The combination of color Doppler sonography and sonoelastography in addition to B-mode sonography leads to an increased accuracy in distinguishing benign from malignant breast masses and to an improvement in characterization of fibroadenomas independent of their shape or size. Show more

Keywords: Fibroadenoma, ultrasound, elastography, doppler sonography

DOI: 10.3233/CH-141884

Citation: Clinical Hemorheology and Microcirculation, vol. 58, no. 1, pp. 115-125, 2014

Authors: Dietze, Kathrin | Slosarek, Ilka | Fuhrmann-Selter, Tania | Hopperdietzel, Carsten | Plendl, Johanna | Kaessmeyer, Sabine

Article Type: Research Article

Abstract: Arterial or venous thromboses are frequent clinical complications with the risk of fatal progression. Recent studies suggest the disruption of angiogenesis in the course of thrombus resolution as the underlying pathomechanism. Very similar to the situation in human patients, equine vessels have been described to be particularly susceptible to thrombosis. In contrast to humans, equine donors are readily available to obtain organs and tissues for isolation of endothelial cells. Objective of this study was to isolate equine endothelial cells and develop an angiogenesis assay from primary cultures. Macrovascular endothelial cells were obtained from jugular veins and carotid arteries of nine …horses, one of which suffered from inflammatory processes. After enzymatic isolation, the cells were incubated in different selective primary media. Phenotypic identification of endothelial cells was accomplished by morphology and positive staining to von Willebrand factor. The reliable, inexpensive, and standardized combination of methods presented here resulted in pure endothelial cultures for angiogenesis assays that can be used in any cell culture laboratory. Inverted phase microscopy and life cell imaging was used to characterize the stages of the angiogenic cascade of the endothelial cells. Life cell imaging gave new insights into the in vitro formation of capillary like structures including exocytosis of microparticles from endothelial cells before integration into the three-dimensional structure. We hypothesize that a specific population of endothelial cells showing a highly active migration pattern in life cell imaging might play a role in the resolution of thrombosis. Show more

Keywords: Equine macrovascular endothelial cells, life cell imaging, angiogenesis assay, microparticles

DOI: 10.3233/CH-141877

Citation: Clinical Hemorheology and Microcirculation, vol. 58, no. 1, pp. 127-146, 2014

Authors: Roch, Toralf | Akymenko, Oksana | Krüger, Anne | Jung, Friedrich | Ma, Nan | Lendlein, Andreas

Article Type: Research Article

Abstract: The polarization behavior of macrophages determines the clinical outcome after implantation of biomaterials. Formation of classically activated macrophages (CAM) may result in cell fusion to form foreign body giant cells, which induce and support uncontrolled inflammatory responses and can cause undesired material degradation. In contrast, polarization into alternatively activated macrophages (AAM) is assumed to support healing processes and implant integration. The expression of matrix metalloproteinases (MMP) by the different macrophage subsets might play a crucial role for inflammatory and wound healing processes and may subsequently influence the implant integration. Therefore, it is of importance to characterize the MMP expression pattern …by the different macrophage subsets. This knowledge could support the design of biomaterials in which specific MMP cleavage sites are incorporated allowing a controlled cell-mediated degradation of the material. However, it needs to be considered that the pure expression levels may not correlate with the enzymatic activity of the MMP, which depends on a variety of different parameters such as additional co-factors. For this reason, the differential MMP expression levels and the overall enzymatic activity of in vitro generated human non-polarized macrophages (M0), CAM, and AAM are analyzed in this study. While MMP-1, MMP-3, and MMP-10 showed the highest expression levels in CAM, MMP-12 was most strongly expressed by AAM. Interestingly, although various MMP were expressed at high levels in CAM, the enzymatic MMP activity was increased in supernatants of AAM cultures. The data presented here illustrate the importance to combine the measurement of MMP expression levels with the analysis of the enzymatic activity. The observed MMP-12 expression in combination with the higher enzymatic activity detected in AAM supernatants might motivate the design of biomaterials, whose structure could be modified by MMP-12 catalyzed reactions leading to interactive polymers. Show more

Keywords: Biomaterials, macrophage polarization, matrix metalloproteinase

DOI: 10.3233/CH-141885

Citation: Clinical Hemorheology and Microcirculation, vol. 58, no. 1, pp. 147-158, 2014

Authors: Braune, S. | Walter, M. | Schulze, F. | Lendlein, A. | Jung, F.

Article Type: Research Article

Abstract: For in vitro studies assessing the interaction of platelets with implant materials, common and standardized protocols for the preparation of platelet rich plasma (PRP) are lacking, which may lead to non-matching results due to the diversity of applied protocols. Particularly, the aging of platelets during prolonged preparation and storage times is discussed to lead to an underestimation of the material thrombogenicity. Here, we study the influence of whole blood- and PRP-storage times on changes in platelet morphology and function. Blood from apparently healthy subjects was collected according to a standardized protocol and examined immediately after blood collection, four hours and …twenty four hours later. The capability of platelets to adhere and form stable aggregates (PFA100, closure time) was examined in sodium citrate anticoagulated whole blood (WB) using the agonists equine type I collagen and epinephrine bitartrate (collagen/epinephrine) as well as equine type I collagen and adenosine-5′-diphosphate (collagen/ADP). Circulating platelets were quantified at each time point. Morphology of platelets and platelet aggregates were visualized microscopically and measured using an electric field multi-channel counting system (CASY). The percentage of activated platelets was assessed by means of P-selectin (CD62P) expression of circulating platelets. Furthermore, platelet factor 4 (PF4) release was measured in platelet poor plasma (PPP) at each time point. Whole blood PFA100 closure times increased after stimulation with collagen/ADP and collagen/epinephrine. Twenty four hours after blood collection, both parameters were prolonged pathologically above the upper limit of the reference range. Numbers of circulating platelets, measured in PRP, decreased after four hours, but no longer after twenty four hours. Mean platelet volumes (MPV) and platelet large cell ratios (P-LCR, 12 fL – 40 fL) decreased over time. Immediately after blood collection, no debris or platelet aggregates could be visualized microscopically. After four hours, first debris and very small aggregates occurred. After 24 hours, platelet aggregates and also debris progressively increased. In accordance to this, the CASY system revealed an increase of platelet aggregates (up to 90 μm diameter) with increasing storage time. The percentage of CD62P positive platelets and PF4 increased significantly with storage time in resting PRP. When soluble ADP was added to stored PRP samples, the number of activatable platelets decreased significantly over storage time. The present study reveals the importance of a consequent standardization in the preparation of WB and PRP. Platelet morphology and function, particularly platelet reactivity to adherent or soluble agonists in their surrounding milieu, changed rapidly outside the vascular system. This knowledge is of crucial interest, particularly in the field of biomaterial development for cardiovascular applications, and may help to define common standards in the in vitro hemocompatibility testing of biomaterials. Show more

Keywords: Platelet, platelet function, platelet rich plasma, whole blood, platelet aging, platelet storage, hemocompatibility, biomaterials

DOI: 10.3233/CH-141876

Citation: Clinical Hemorheology and Microcirculation, vol. 58, no. 1, pp. 159-170, 2014

Authors: Gerk, U. | Krüger, A. | Franke, R.P. | Jung, F.

Article Type: Research Article

Abstract: Radiographic contrast media can lead to drastic changes of the morphology of erythrocytes. The change of the erythrocyte morphology is associated with a decreased deformability possibly resulting from distinctions in the loss of constituents of the membrane cytoskeleton. However, it is unclear whether there is an intravascular hemolysis as a consequence of the disintegration of the erythrocyte membrane. The results of this study showed, that free haemoglobin increased from 16.8 ± 10.0 mg/dl to 21.6 ± 12.6 mg/dl after Iopromide application (p = 0.240), while it slightly decreased from 20.5 ± 10.3 mg/dl to 19.5 ± 12.2 mg/dl after Iodixanol …application (p = 0.547). The slight decrease of free haemoglobin after application of Iodixanol differed significantly compared to the increase of free haemoglobin after Iopromide application (p < 0.05). This different response is thought to give evidence to the assumption that the erythrocyte membrane integrity was compromised leading to the release of free haemoglobin as an indicator of hemolysis as well. Show more

Keywords: Radiographic contrast media, Iopromide, Iodixanol, hemolysis, free haemoglobin

DOI: 10.3233/CH-141879

Citation: Clinical Hemorheology and Microcirculation, vol. 58, no. 1, pp. 171-174, 2014

Authors: Gori, Tommaso | Münzel, Thomas

Article Type: Research Article

Abstract: Coronary artery stenting is associated with endothelial dysfunction, probably caused by the mechanical injury as well as by the tissue reactions triggered by the metal, the polymer and the drug eluted by the stent. The clinical relevance, and the implications, of this dysfunction are still unclear. We revise the iterature on this topic and provide the first evidence regarding endothelial dysfunction after implantation of bioresorbable scaffolds.

Keywords: Endothelium/vascular type/nitric oxide, pathophysiology, coronary circulation

DOI: 10.3233/CH-141880

Citation: Clinical Hemorheology and Microcirculation, vol. 58, no. 1, pp. 175-181, 2014

Authors: Teusch, V.I. | Wohlgemuth, W.A. | Piehler, A.P. | Jung, E.M.

Article Type: Research Article

Abstract: AIM: Aim of our pilot study was the application of a contrast-enhanced color-coded ultrasound perfusion analysis in patients with vascular malformations to quantify microcirculatory alterations. MATERIAL AND METHODS: 28 patients (16 female, 12 male, mean age 24.9 years) with high flow (n = 6) or slow-flow (n = 22) malformations were analyzed before intervention. An experienced examiner performed a color-coded Doppler sonography (CCDS) and a Power Doppler as well as a contrast-enhanced ultrasound after intravenous bolus injection of 1 – 2.4 ml of a second-generation ultrasound contrast medium (SonoVue® , Bracco, Milan). The contrast-enhanced examination was documented as a cine …sequence over 60 s. The quantitative analysis based on color-coded contrast-enhanced ultrasound (CEUS) images included percentage peak enhancement (%peak), time to peak (TTP), area under the curve (AUC), and mean transit time (MTT). RESULTS: No side effects occurred after intravenous contrast injection. The mean %peak in arteriovenous malformations was almost twice as high as in slow-flow-malformations. The area under the curve was 4 times higher in arteriovenous malformations compared to the mean value of other malformations. The mean transit time was 1.4 times higher in high-flow-malformations compared to slow-flow-malformations. There was no difference regarding the time to peak between the different malformation types. The comparison between all vascular malformation and surrounding tissue showed statistically significant differences for all analyzed data (%peak, TTP, AUC, MTT; p < 0.01). High-flow and slow-flow vascular malformations had statistically significant differences in %peak (p < 0.01), AUC analysis (p < 0.01), and MTT (p < 0.05). CONCLUSION: Color-coded perfusion analysis of CEUS seems to be a promising technique for the dynamic assessment of microvasculature in vascular malformations. Show more

Keywords: Contrast-enhanced ultrasound (CEUS), vascular malformations, color-coded perfusion analysis, microcirculation

DOI: 10.3233/CH-141878

Citation: Clinical Hemorheology and Microcirculation, vol. 58, no. 1, pp. 183-193, 2014

Authors: Trottmann, M. | Marcon, J. | D'Anastasi, M. | Karl, A. | Stief, C.G. | Reiser, M. | Clevert, D.A.

Article Type: Research Article

Abstract: PURPOSE: Virtual touch tissue imaging quantification (VTIQ) is a newly developed technique for the sonographic quantification of tissue elasticity. It has been used in the assessment of breast lesions. The purpose of this study was to determine the diagnostic performance of VTIQ in indeterminate testicular lesions. METHODS: Twenty patients with known testicular pathology underwent conventional B-mode sonography with additional VTIQ of the testicular lesions using a Siemens Acuson S2000™ and S3000™ (Siemens Medical Solutions, Mountain View, CA, USA) system. Tissue mechanical properties were analysed in the VTIQ examination. The pathologic diagnosis was established after surgery or in the follow-up examination …for suspected benign lesions. RESULTS: Over 36 months, 22 focal testicular lesions (median lesion size, 18 mm; range, 4–36 mm in 20 patients (median age, 43 years; range, 22–81 years) were examined. Lesions were hyperechoic (n = 1), hypoechoic (n = 14), isoechoic (n = 1), of mixed echogenicity (n = 3) or anechoic (n = 3). Histological examination showed one benign lesion (6.25%) with a mean size of 7 mm and 15 malignant lesions (93.75%) with a mean size of 20 mm. Mean shear wave velocity for normal testicular tissue was 1.17 m/s. No shear wave velocity could be measured in cystic lesions. The rest of the benign lesions showed a mean shear wave velocity of 2.37 m/s. The value of the shear wave velocity in germ cell tumours showed a mean shear wave velocity of 1.94 m/s and for seminoma it showed a mean shear wave velocity of 2.42 m/s. CONCLUSIONS: VTIQ is a reliable new method for measuring qualitative and quantitative stiffness of testicular lesions and tissue. The qualitative shear-wave elastography features were highly reproducible and showed good diagnostic performance in unclear testicular lesions. The VTIQ technique is also useful in assessing small testicular nodules and pseudolesions. Show more

Keywords: Elastography, shear wave imaging, seminoma, germ cell tumour, testis

DOI: 10.3233/CH-141904

Citation: Clinical Hemorheology and Microcirculation, vol. 58, no. 1, pp. 195-209, 2014

Authors: George, S. | Georgi, M. | Roggenbuck, D. | Conrad, K. | Küpper, J.-H.

Article Type: Research Article

Abstract: Many autoimmune diseases are characterized by autoantibodies directed against cell membrane proteins. We were intrigued to develop a strategy for targeting individual cell membrane proteins to various subcellular compartments as a prerequisite for their simultaneous immunofluorescence detection. We first employed GFP and RFP reporters that were equipped with defined intracellular localization signals. Expressing these protein reporters in HEp-2 cells we found by using fluorescence microscopy that protein localization in cytoplasm or at mitochondria can be clearly discriminated from localization at Golgi, ER or lysosomes. We then tested for muscle-specific kinase, a relevant cell membrane autoantigen in Myasthenia gravis, and NMDA …receptor which is relevant for autoimmune encephalitis, whether these autoantigens can be localized to the same intracellular compartments. To this end, we successfully targeted muscle-specific kinase to Golgi apparatus, mitochondria and cytoplasm. We found that its Golgi localization can be clearly distinguished from its natural cell membrane localization. The same we found for Golgi-localized NMDA receptor 1. Interestingly, cell membrane proteins kept at the Golgi system accumulated in higher amounts than their wild-type counterparts. The obtained results are the basis for the further development of multiplex assays for the immunofluorescence diagnostics of Myasthenia gravis and autoimmune encephalitis. Show more

Keywords: AKLIDES, autoimmune diseases, encephalitis, immunofluorescence, intracellular trafficking, multiplex diagnostics, Myasthenia gravis

DOI: 10.3233/CH-141897

Citation: Clinical Hemorheology and Microcirculation, vol. 58, no. 1, pp. 211-228, 2014

Authors: Krüger, Anne | Mayer, Anke | Roch, Toralf | Schulz, Christian | Lendlein, Andreas | Jung, Friedrich

Article Type: Research Article

Abstract: Angiogenically stimulated alternative monocytes (aMO2) could be established as cellular release system accelerating the endothelialization of polymers rendering their surfaces hemocompatibility in a short-term study. However, for their clinical application it is essential that aMO2 do not switch back to the MO1 state sustaining their capability as cellular release system over an extended period of time. We explored whether aMO2 can maintain their differentiation state over 21 days in a mono- and in a co-culture with HUVEC. In comparison, the influence of recombinant VEGF-A165 on the endothelialization of biomaterials was assessed including endothelial cell (HUVEC) density, organisation of the …endothelial cytoskeleton, cytokine secretion profile and release of prostacyclin, thromboxane A2 and matrix metalloproteinases. In mono-culture aMO2 secreted high amounts of VEGF and other growth factors/cytokines. Co-cultured with HUVEC, aMO2 accelerated the formation of a confluent HUVEC monolayer. Furthermore, no pro-inflammatory cytokines were found, neither in aMO2-mono, nor in co-cultures with HUVEC indicating that the majority of the aMO2 remained stable in their aMO2 state during the 21 days of cultivation. In contrast, the addition of recombinant VEGF-A165 instead of the co-culture with aMO2 resulted in the formation of stress fibres, dissociated marginal filament bands, and a detachment of HUVEC. In addition, the profile of bioactive agents of HUVEC (e.g. prostacyclin, thromboxane A2, matrix metalloproteinases, IFN-γ and TNF-α) was influenced by the VEGF-A165 treatment inducing the detachment of HUVEC. In conclusion, in co-culture with HUVEC aMO2 remained stable in their type 2 state over 21 days confirming the suitability of aMO2 as biological release system for the endothelialization of biomaterial surfaces with constant release of angiogenic factors but without secretion of pro-inflammatory cytokines over three weeks. Therefore, this endothelialization approach seems to be appropriate to improve the hemocompatibility of cardiovascular implant materials in vitro, and proved to be superior to the use of recombinant VEGF-A165 . Show more

Keywords: Angiogenically stimulated alternative monocytes, endothelialization, biomaterials, hemocompatible surface

DOI: 10.3233/CH-141875

Citation: Clinical Hemorheology and Microcirculation, vol. 58, no. 1, pp. 229-240, 2014

Authors: Manz, Patrick | Cadeddu, Ron-Patrick | Wilk, Matthias | Fritz, Birgit | Haas, Rainer | Wenzel, Folker

Article Type: Research Article

Abstract: INTRODUCTION: Softeners like phthalate esters are ubiquitous in the environment and have been detected in transfusion bags, though there is only a limited amount of studies on the effect of phthalates on blood cells. This study seeks to determine effects on cell migration of human promyelocytic leukemia cells (HL-60) incubated with di(2-ethylhexyl)phthalate (DEHP) at concentrations found in blood bags. MATERIAL AND METHODS: HL-60 cells were incubated with DEHP concentrations ranging from 0.1 μg/ml to 1000 μg/ml, diluted in DMSO, over 24 h, 48 h, and 72 h. Migration rate was analyzed along an SDF-1α gradient using Transwell migration inserts. RESULTS: …Of the applied concentrations 100 μg/ml, 250 μg/ml, 500 μg/ml, and 1000 μg/ml showed a significant decrease in migration rates relative to DMSO control at all measuring points (p < 0.05), with relative migration rates between 37.87 % for 100 μg/ml and 25.34 % for 1000 μg/ml relative to DMSO after 24 h of stimulation and 19.73 % for 100 μg/ml and 14.69 % for 1000 μg/ml respectively after 72 h of incubation. CONCLUSION: Our results indicate HL-60 to be a suitable in vitro model for examining effects of DEHP on the migration of blood and nucleated cells at concentrations found in blood bags. Show more

Keywords: Phthalates, DEHP, MEHP, HL-60 promyelocytes, vitality, migration, apoptosis

DOI: 10.3233/CH-141903

Citation: Clinical Hemorheology and Microcirculation, vol. 58, no. 1, pp. 241-246, 2014

Authors: Zimmermann, H. | D'Anastasi, M. | Rjosk-Dendorfer, D. | Helck, A. | Meimarakis, G. | Reiser, M. | Clevert, D.A.

Article Type: Research Article

Abstract: PURPOSE: To compare the value of high-resolution contrast-enhanced ultrasound in the detection and characterisation of endoleaks after EVAR. MATERIAL AND METHODS: In this retrospective study, a cohort of 80 patients underwent both standard examination using a curved array 5 MHz transducer and a second examination using a curved array 9 MHz transducer. The examination included B-mode, color Doppler and contrast-enhanced ultrasound using the Philips EPIQ 7 scanner. RESULTS: 80 patients were included in the study. CEUS was used as the preferred examination in determining the presence of an endoleak. The sensitivity and specificity for the detection of endoleak using the …5 MHz transducer was: CEUS (96.8%, 100%), Color Doppler (47.6%, 94.1%), B-mode (15.8%, 94.1%), while for the 9 MHz transducer: CEUS (100%, 100%), Color Doppler (39.7%, 94.1%), B-mode (23.8%, 24.1%). CONCLUSION: High resolution ultrasound (9 MHz) in detection and characterization of endoleaks after EVAR has some minor advantages in comparison to lower resolution ultrasound (5 MHz). Show more

Keywords: Endoleak, EVAR, AAA, CEUS, high resolution ultrasound

DOI: 10.3233/CH-141896

Citation: Clinical Hemorheology and Microcirculation, vol. 58, no. 1, pp. 247-260, 2014

Authors: Klotz, Laura V. | Ingrisch, Michael | Eichhorn, Martin E. | Niemoeller, Olivier | Siedek, Vanessa | Gürkov, Robert | Clevert, Dirk-Andre

Article Type: Research Article

Abstract: PURPOSE: Contrast enhanced ultrasound (CE-US) is a promising imaging modality for non-invasive analysis of functional vascularisation. Lesions of the parotid gland are associated with a vascularisation that differs from normal gland tissue. The aim of this clinical study was to further analyse the perfusion in parotid gland lesions with CE-US. The new quantification software VueBox (Bracco, Italy) was used to assess the perfusion, based on DICOM datasets of CE-US examination. MATERIALS AND METHODS: CE-US measurements were performed by intravenous application of a contrast agent (SonoVue, Bracco, Italy) before surgical tumor resection. From the analysis of a time sequence of 2D …DICOM contrast images, area under time intensity curve (AUC), peak enhancement (PE), wash-in-rate (WiR) and wash-in-perfusion-index (WiPI) were calculated using VueBox. These were correlated with the histological analyses of the tumor tissue. RESULTS: Significant difference of area below intensity time curve (AUC), peak enhancement (PE), wash-in-rate (WiR) and wash-in perfusion index (WiPI) were observed in the malign lesions compared to benign tumors (p < 0,05) and in pleomorphic adenoma compared to cystadenolymphoma (p < 0,05). CONCLUSION: CE-US seems to be a quantitative and independent method for discriminating between malign and benign parotid gland tumors. Show more

Keywords: Parotid gland, contrast enhanced ultrasound, tumor microcirculation, salivary gland, tumor, perfusion, imaging

DOI: 10.3233/CH-141895

Citation: Clinical Hemorheology and Microcirculation, vol. 58, no. 1, pp. 261-269, 2014

Authors: Weinrich, M. | Schindler, P. | Kundt, G. | Klar, E. | Bünger, C.M.

Article Type: Research Article

Abstract: BACKGROUND: For the reduction of cardio- and cerebrovascular events in carotid endarterectomies continuation of antiplatelet medication is recommended perioperatively. As a result, this patient population is at increased risk for postoperative bleeding complications. Intraoperative application of local hemostatic agents might reduce the incidence of bleeding complications. MATERIAL AND METHODS: All 565 patients undergoing carotid endarterectomy between January 2005 and January 2011 were analysed retrospectively. Most patients in the earlier cohort years of the study had no perioperative antiplatelet medication. In contrast antiplatelet medication was usually continued perioperatively in the later cohort years. To reduce the risk of perioperative bleeding local …hemostatic agents were applied increasingly. RESULTS: Revision surgery, due to postoperative bleeding or massive hematoma, was necessary in 20 cases (3.5 %). Overall, 383 carotid endarterectomies (67.8 %) were performed with perioperative antiplatelet medication. Local hemostatic agents were applied in 259 cases (45.8 %) intraoperatively. Initially, operations performed in patients taking antiplatelet medication resulted in an increased need for surgical revision. Following an accelerated practice of using local hemostatic agents, the need for revision surgeries fell. Nevertheless, when patients from all years were analysed together there was no significant benefit from the application of local hemostatic agents. CONCLUSION: Application of local hemostatic agents might have contributed to a reduction of bleeding complications in carotid endarterectomies. However, this could not be shown of statistical significance. Other confounding factors such as different operative techniques or forms of anesthesia might also have influenced this decline. Show more

Keywords: Hemostatic agents, carotid endarterectomy, antiplatelet medication, bleeding complication

DOI: 10.3233/CH-141901

Citation: Clinical Hemorheology and Microcirculation, vol. 58, no. 1, pp. 271-279, 2014

Authors: Ullm, Sandra | Laube, Markus | Bechmann, Nicole | Kniess, Torsten | Pietzsch, Jens

Article Type: Research Article

Abstract: Radiotherapy of various cancers is closely associated with increased cardiovascular morbidity and mortality. Arachidonic acid metabolites are supposed to play a key role in radiation-induced vascular dysfunction. This study was designed to evaluate the effects of novel, antioxidative 2,3-diaryl-substituted indole-based selective cyclooxygenase-2 (COX-2) inhibitors (2,3-diaryl-indole coxibs) on radiation-induced formation of arachidonic acid metabolites via COX-2 and oxidant stress pathways in an organotypical vascular model of rat aortic rings. Acute and subacute effects of X-ray radiation (4 and 10 Gy; 1 and 3 days post irradiation) with or without the presence of 1 μM of the 2,3-diaryl-indole coxib 2-[4-(aminosulfonyl)phenyl]-3-(4-methoxyphenyl)-1H-indole (C1) or …celecoxib as reference compared to sham-irradiated controls were assessed. The following parameters were measured: metabolic activity of the aortic rings; induction and regulation of COX-2 expression; release of prostaglandin E2 and F2α -isoprostane. Irradiation without presence of coxibs resulted in a dose-dependent augmentation of all parameters studied. When aortic rings were exposed to the 2,3-diaryl-indole coxib 1 h before irradiation, metabolic activity was restored and the release of both prostaglandin and isoprostane was inhibited. The latter indicates a direct interaction with oxidant stress pathways. By contrast, celecoxib exhibited only slight effects on the formation of isoprostane. The reduction of radiation-induced vascular dysfunction by antioxidative coxibs may widen the therapeutic window of COX-2 targeted treatment. Show more

Keywords: Aortic ring model, coxibs, prostanoids, normal tissue damage, radiation therapy, adjuvant radioprotective therapy, cardiovascular disease

DOI: 10.3233/CH-141902

Citation: Clinical Hemorheology and Microcirculation, vol. 58, no. 1, pp. 281-295, 2014