bluuebunny/crossref_metadata_2025 huggingface dataset:
Polyphenols of leaves of Apium graveolens inhibit in vitro protein glycation and protect RINm5F cells against methylglyoxal-induced cytotoxicity
Authors: Perez Gutierrez Rosa Martha , Muñiz-Ramirez Alethia , Campoy Abraham Heriberto Garcia , Flores Jose Maria Mota , Flores Sergio Odin
Published: December 2019

Background: The health benefits of edible plants have been widely investigated and disseminated. However, only polyphenols have been found to have sufficient therapeutic potential to be considered in clinical trials. Fewer manuscripts have other applications such as prospective health benefits and disease treatment. Other components of edible plants are responsible for a range of other benefits including antimalarial, burns, flu, cancer, inflammation, diabetes, glycation, antimicrobial, prevention of neurodegeneration, analgesic, antimigraine activity, sedative activities, etc. Accordingly, the public needs to be informed of the potential edible plants have to act on different targets and maintain better control over diabetes compared to commercial drugs which can be toxic, have side effects, do not have the capacity to maintain blood glucose at normal levels, and do not protect the patient from the complications of diabetes over time. Consequently, edible plants, such as Apium graveolen, which have therapeutic targets on AGEs formation, are potentially a better alternative treatment for diabetes.Methods: The leaves of celery were extracted with methanol (CM). Polyphenols contents in CM were investigated by liquid chromatography-electrospray ionization mass. The ability of the compounds to inhibit formation of AGEs was evaluated in vitro models using formation of AGE fluorescence intensity, level of fructosamine, Nε-(carboxymethyl)lysine (CML), methylglyoxal (MG)-derived protein, and formation of amyloid cross β structure. Protein-oxidation was determined by thiol group and protein carbonyl content. Inhibition of MG-derived AGEs and MG-trapping ability were also measured. Additionally, insulin production was determined in methylglyoxal-treated pancreatic RINm5F cells assay. Results: Apigenin, kaempferol, apiin, rutin, caffeic acid, ferulic acid, chlorogenic acid, coumaroylquinic acid, and p-coumaric acid were the major polyphenols contained in CM. In all the model tests CM displayed potent AGE inhibitory activity, suggesting that CM delayed the three stages of glycation. Accordingly, the mechanisms of action of celery involving dicarbonyl trapping and breaking the crosslink structure in the AGEs formed may contribute to the protection of pancreatic RINm5F cells against MG conditions.Conclusion: These findings indicate that CM have an excellent anti-glycation effect which may be beneficial for future development of antiglycating agents for the treatment of diabetes.Keywords: Apium graveolens, anti-glycation, polyphenols methylglyoxal, insulin, pancreatic cells
Anti-obesity and haematological effects of Malaysia Hibiscus sabdariffa L. aqueous extract on obese Sprague Dawley rats
Authors: Omar Maizatul Hasyima , Shamsahal Norsyafawati , Muhammad Hussin , Wan Ahmad Wan Amir Nizam , Wasiman Mohd Isa
Published: December 2019

Background: Hibiscus sabdariffa L. (Hs; Malvaceae) is commonly known as roselle or red sorrel in English or karkadè in Arabic. It is a tropical plant native to India and Malaysia. Roselle extracts have been widely reported to have beneficial health effects. The aim of this study was to determine the effective dose of Hs aqueous extract, which is a possible reducing agent in diet-induced obese rats, and monitoring any toxicological effect.Methods: Male Sprague Dawley rats (n= 24) aged 6-7 weeks and weighing 210 ± 3.5 g (mean ± S.E.M) were used.  The rats were fed a high fat diet (HFD) for 8 consecutive weeks to induce obesity. The animals were then randomly assigned to one of five groups based on the concentration of Hs aqueous extract. The normal group (NG) received a normal diet (commercial chow) throughout the study. The obese group (ObG) included obese rats administered with tap water ad libitum. Groups 1 (150 mg/kg), 2 (200 mg/kg), 3 (250 mg/kg), and 4 (300 mg/kg) were obese rats continuously fed with HFD in combination with the Hs extract for 10 weeks. The effect of Hs on some haematological and blood biochemical parameters were also evaluated.Results:  Hs aqueous extract is a rich source of anthocyanins. The main compounds detected are delphinidin-3-O-sambubioside and delphinidin-3-O-sambubioside. The extract also possesses high antioxidant properties which may be caused by these anthocyanins. The anti-obesity effect of Hs aqueous extract was demonstrated by the significant reduction in the weight gain and abdominal weight (p < 0.05) between treated and non-treated groups, which was dose-dependent. Overall, our study demonstrated that oral administration of Hs at doses of 150, 200, 250, and 300 mg/kg for ten weeks did not cause any toxicity effect within the obese rats.Conclusions: The results of the study implied Hs aqueous extract at 300 mg/kg is the dose which can the most weight reduction effect with no severe haematological and biochemical changes in all experimental animals.Keywords: Hibiscus sabdariffa, obese rats, roselle, aqueous extract, and body weight
T lymphocytes alterations are associated with oxidized LDL, troponin T, white blood cells and C-reactive protein during acute myocardial infarction
Authors: Napoleão Patrícia , Selas Mafalda , Freixo Cláudia , Carmo Miguel Mota , Viegas-Crespo Ana Maria , Ferreira Rui Cruz , Pinheiro Teresa
Published: December 2019

BACKGROUND: Auto-immune responses are associated with oxidized LDL (ox-LDL) release, a key factor in plaque destabilization. Data on the relationship between ox-LDL and T lymphocytes in human populations remains scarce. T cells also react with other molecules from the lesion and/or damage the myocardium. OBJECTIVE: The objective of the present study was to examine the relationship between circulating T lymphocytes, ox-LDL, markers of myocardial necrosis (cTnT), myocardial dysfunction (N-terminal pro-brain natriuretic peptide – NT-proBNP) and inflammation (C-reactive protein – CRP) in the setting of acute myocardial infarction. METHODS: A longitudinal study of 55 patients with ST-elevation myocardial infarction (STEMI) were evaluated at three time points: admission, 2 and 40 days following admission, together with 30 patients with stable angina (SA) and 56 subjects without coronary artery disease serving as controls (CTR). RESULTS: STEMI patients had maximal ox-LDL values and minimal levels of CD3+ T lymphocytes at admission, which was normalized during the recovery period. The increasing trend of CD3+ T cells was positively associated with an ox-LDL decline over time. CRP and cTnT longitudinal variations were negatively associated with the CD3+ T-cell increasing trend. These associations were not found in SA patients or controls. CONCLUSIONS: The associations found between CD3+ T lymphocytes, ox-LDL and cTnT suggest a specificity of the immune response in AMI towards arterial and myocardial inflammation and remodelling.
Hemorheological changes in ischemia-reperfusion: An overview on our experimental surgical data
Authors: Nemeth Norbert , Furka Istvan , Miko Iren
Published: December 2019

Blood vessel occlusions of various origin, depending on the duration and extension, result in tissue damage, causing ischemic or ischemia-reperfusion injuries. Necessary surgical clamping of vessels in vascular-, gastrointestinal or parenchymal organ surgery, flap preparation-transplantation in reconstructive surgery, as well as traumatological vascular occlusions, all present special aspects. Ischemia and reperfusion have effects on hemorheological state by numerous ways: besides the local metabolic and micro-environmental changes, by hemodynamic alterations, free-radical and inflammatory pathways, acute phase reactions and coagulation changes. These processes may be harmful for red blood cells, impairing their deformability and influencing their aggregation behavior. However, there are still many unsolved or non-completely answered questions on relation of hemorheology and ischemia-reperfusion. How do various organ (liver, kidney, small intestine) or limb ischemic-reperfusionic processes of different duration and temperature affect the hemorheological factors? What is the expected magnitude and dynamics of these alterations? Where is the border of irreversibility? How can hemorheological investigations be applied to experimental models using laboratory animals in respect of inter-species differences? This paper gives a summary on some of our research data on organ/tissue ischemia-reperfusion, hemorheology and microcirculation, related to surgical research and experimental microsurgery.
Predictive value of brachial reactive hyperemia and flow-mediated dilation in stable coronary artery disease
Authors: Ostad Mir Abolfazl , Wild Philipp S. , Schnorbus Boris , Schulz Andreas , Munzel Thomas , Warnholtz Ascan
Published: December 2019

BACKGROUND: The purpose of this study was to determine the predictive value of a single measurement of reactive hyperemia (RH) and brachial flow-mediated dilation (FMD) in patients with established stable coronary artery disease (CAD). METHODS: RH and brachial artery FMD were ultrasonographically measured in 325 patients with stable CAD. Patients were followed for cerebro-cardiovascular events. The median follow-up was 3.7 years (range 0.01–5.7 years). RESULTS: Sixty-seven patients (20.6%) had an cerebro-cardiovascular event. Patients with subsequent events had lower FMD (4.9 ± 3.3% versus 6.3 ± 3.5%, p = 0.003), higher brachial artery resting diameter (5.1 ± 0.7 mm versus 4.8 ± 0.7 mm, p = 0.002) and lower NMD (11.2 ± 5.1% versus 12.8 ± 5.4%, p = 0.02), while the mean hyperemic flow velocity and shear stress did not differ from patients without cerebro-cardiovascular events. Cox proportional hazard model adjusted for sex, age, BMI, and traditional cardiovascular risk factors revealed a hazard ratio of 0.84 for lower FMD (p = 0.01). CONCLUSIONS: We conclude that single spot measurements of peak RH do not provide long-term prognostic information, but evaluation of conduit artery FMD predicts long-term cerebro-cardiovascular events in patients with stable CAD. The prognostic value of FMD is incremental to traditional cardiovascular risk factors and may therefore be of clinical importance.
Indirect viscosimetric method is less accurate than ektacytometry for the measurement of red blood cell deformability
Authors: Vent-Schmidt Jens , Waltz Xavier , Pichon Aurélien , Hardy-Dessources Marie-Dominique , Romana Marc , Connes Philippe
Published: December 2019

The aim of this study was to test the accuracy of viscosimetric method to estimate the red blood cell (RBC) deformability properties. Thirty-three subjects were enrolled in this study: 6 healthy subjects (AA), 11 patients with sickle cell-hemoglobin C disease (SC) and 16 patients with sickle cell anemia (SS). Two methods were used to assess RBC deformability: 1) indirect viscosimetric method and 2) ektacytometry. The indirect viscosimetric method was based on the Dintenfass equation where blood viscosity, plasma viscosity and hematocrit are measured and used to calculate an index of RBC rigidity (Tk index). The RBC deformability/rigidity of the three groups was compared using the two methods. Tk index was not different between SS and SC patients and the two groups had higher values than AA group. When ektacytometry was used, RBC deformability was lower in SS and SC groups compared to the AA group and SS and SC patients were different. Although the two measures of RBC deformability were correlated, the association was not very high. Bland and Altman analysis demonstrated a 3.25 bias suggesting a slight difference between the two methods. In addition, the limit of agreement represented 28% (>15%) of the mean values of RBC deformability, showing no interchangeability between the two methods. In conclusion, measuring RBC deformability by indirect viscosimetry is less accurate than by ektacytometry, which is considered the gold standard.
Effect of radiographic contrast media (Iodixanol, Iopromide) on the spectrin/actin-network of the membranous cytoskeleton of erythrocytes
Authors: Franke R.P. , Scharnweber T. , Fuhrmann R. , Mrowietz C. , Jung F.
Published: December 2019

Red blood cells demonstrate a unique ability for repeated large deformation. Under the influence of a variety of agents, shapes other than the discocyte – e.g. stomatocytes or echinocytes - can be observed. Some radiographic agents induce shape changes from discocytic to echinocytic cells. Especially the echinocyte formation is associated with a rigidification of the cells bearing the risk of a hindered capillary passage of the echinocytes. The mechanisms leading to the formation of echinocytes are not well understood assuming that the membrane cytoskeleton is a key player. That is why this examination was focused on the participation of components of the membrane cytoskeleton in the formation of echinocytes and the protrusions accompanying the formation of echinocytes. Two radiographic contrast media approved for intra-arterial application were used to study echinocyte formation (Iodixanol320; Iopromide370). In the in vitro study serious changes in the membrane cytoskeleton were only found in those erythrocytes incubated in plasma supplemented with Iopromide370 (30%v/v). The shape of the spectrin net was completely altered; from the more homogeneous distribution - typical of cells in autologous plasma and also of cells in plasma supplemented with Iodixanol320 – to a distribution of spectrin concentrated in the membrane-near regions with the appearance of spectrin-actin co-localization. Co-localized spectrin with actin was also found around the membranous roots of protrusions which resemble exocytotic processes. In central parts of the cells there was a pronounced dissociation of spectrin and actin; green coloured condensed spectrin bundles originating from the cell membrane reached up to the root of the protrusions. Separate from this there were also fine long actin fibres passing through the whole cell. The incubation of erythrocytes in plasma supplemented with Iopromide370 induced rounded bubble-like protrusions from the cell membrane containing almost completely long bundles of actin fibres. The examination confirmed earlier studies showing that some radiographic contrast media are able to induce echinocyte formation. Furthermore, subcellular mechanisms were revealed explaining the different effects of Iodixanol in comparison to Iopromide.
Non-invasive pressure measurement of the great saphenous vein in healthy controls and patients with venous insufficiency
Authors: Koster M. , Amann-Vesti B.R. , Husmann M. , Jacomella V. , Meier T.O. , Jeanneret C. , Hafner J. , Thalhammer C.
Published: December 2019

OBJECTIVES: Venous pressure measurement using an intravenous catheter is the sole method for the diagnosis of venous hypertension in patients with chronic venous insufficiency. A noninvasive tool to quantify increased venous pressure is essential for studying venous pathophysiology. Aim of the study was to investigate the value of controlled compression ultrasound (CCU) for noninvasive assessment of venous pressure (VP) of the great saphenous vein (GSV) in healthy persons and patients with venous insufficiency to quantify venous hypertension. METHODS: An optimal visible part of the GSV directly above the ankle was marked on the skin and compressed under ultrasound control and pressure needed for complete compression of the vein was recorded using a pressure manometer with a translucent silicone membrane. Complete insufficiency of the GSV (Hach IV) was documented by duplex ultrasound by an independent investigator before start of the study. VP measurement was performed while normal breathing, deep inspiration and expiration and during a standardized Valsalva maneuver. RESULTS: Twenty controls and 19 patients with complete insufficiency of the GSV were included. Valsalva maneuver induced a slight increase in VP in controls (20.1 ± 4.5 vs 25.1 ± 6.6 mbar) but a significant higher increase in patients from 26 to 37 mbar (IQR 18.5–28.0 vs 31.5–43.0; p < 0.001). CONCLUSION: Noninvasive venous pressure measurement of the great saphenous vein using CCU is feasible and documents an increased pressure during Valsalva maneuver in Hach IV patients compared to healthy controls.
Use of contrast-enhanced ultrasound to monitor rabbit renal ischemia-reperfusion injury and correlations between time-intensity curve parameters and renal ICAM-1 expression
Authors: Li Mingxing , Luo Zhijian , Chen Xiaomei , Xuan Jiqing , Ye Fan , Liu Hui , Chen Kewen
Published: December 2019

BACKGROUND: Renal ischemia-reperfusion injury (IRI) is common in renal transplantation, shock, and nephrolithotomy. OBJECTIVE: To quantitatively assess rabbit renal IRI with contrast-enhanced ultrasound (CEUS) and investigate associations between renal cortical time-intensity curve (TIC) parameters and intracellular adhesion molecule-1 (ICAM-1) expression. METHODS: Eighteen white rabbits were randomly assigned into control and IRI groups (n = 9 each), and CEUS examination was performed 24 h after modeling. TIC parameters including arrival time (AT), time to peak intensity (TTP), changes in peak intensity, area under the curve (AUC), and slope of the ascending TIC, and correlations between these parameters and ICAM-1 expression were evaluated with linear correlation analyses. RESULTS: AT, TTP, and AUC were significantly increased in the IRI group (P < 0.05), and the slope of the ascending TIC was significantly lower than that in the control group (P < 0.05). ICAM-1 content in the renal cortex was significantly increased in the IRI group (P < 0.05). In addition, the slope of the ascending TIC negatively correlated with renal ICAM-1 expression (r = −0.923, P < 0.01), whereas AT and TTP positively correlated with ICAM-1 expression (r = 0.697 and 0.892, both P < 0.01). CONCLUSIONS: TIC parameters including the slope of the ascending TIC, AT, and TTP closely correlated with ICAM-1 in the renal cortex, and it is considered that the TIC can be used to quantitatively monitoring renal cortex blood perfusion and CEUS can be used to indirectly evaluate the degree of inflammatory reaction associated with renal IRI. CEUS may be a useful non-invasive method to identify inflammation caused by renal IRI-associated diseases.
Alpha thalassemia protects sickle cell anemia patients from macro-albuminuria through its effects on red blood cell rheological properties
Authors: Lamarre Yann , Romana Marc , Lemonne Nathalie , Hardy-Dessources Marie-Dominique , Tarer Vanessa , Mougenel Danielle , Waltz Xavier , Tressières Benoît , Lalanne-Mistrih Marie-Laure , Etienne-Julan Maryse , Connes Philippe
Published: December 2019

While chronic hemolysis has been suspected to be involved in the development of glomerulopathy in patients with sickle cell anemia (SCA), no study focused on the implications of blood rheology. Ninety-six adults with SCA at steady state were included in the present cross-sectional study. Three categories were defined: normo-albuminuria (NORMO, n = 41), micro-albuminuria (MICRO, n = 23) and macro-albuminuria (MACRO, n = 32). Blood was sampled to measure hematological and hemorheological parameters, and genomic DNA extraction was performed to detect the presence of α-thalassemia. The prevalence of α-thalassemia was lower in the MACRO group compared with the two other groups. Anemia was more severe in the MACRO compared with the NORMO group leading the former group to exhibit decreased blood viscosity. Red blood cell deformability was lower and red blood cell aggregates strength was greater in the MACRO compared to the two other groups, and this was directly attributed to the lower frequency of α-thalassemia in the former group. Our results show the protective role of α-thalassemia against the development of sickle cell glomerulopathy, and strongly suggest that this protection is mediated through the decrease of anemia, the increase of RBC deformability and the lowering of the RBC aggregates strength.
An effect of glycoprotein IIb/IIIa inhibitors on the kinetics of red blood cells aggregation
Authors: Sokolova Irina A. , Muravyov Alexei V. , Khokhlova Maria D. , Rikova Sofya Yu. , Lyubin Evgeny V. , Gafarova Marina A. , Skryabina Maria N. , Fedyanin Angrey A. , Kryukova Darya V. , Shahnazarov Alexander A.
Published: December 2019

The reversible aggregation of red blood cells (RBCs) continues to be of the basic science and clinical interest. Recently it has been reported about a specific binding between fibrinogen and unknown erythrocyte glycoprotein receptors. The aim of this study was to investigate whether the red blood cell aggregation (RBCA) include the cell-cell interaction using the membrane receptors that bind such ligands as fibrinogen or fibronectin. To test this hypothesis the RBCs were incubated with monafram - the drug of the monoclonal antibodies against glycoprotein (GP) IIb/IIIa, with the GPIIb-IIIa receptor antagonist tirofiban, epifibatide and with the fibrinogen inhibiting peptide. It has been found that the RBC incubation with monafram resulted in a marked RBCA decrease mainly in persons with high level of aggregation. Another research session has shown that RBC incubation with fibronectin was accompanied by a significant RBCA rise. The monafram addition to red cell incubation medium resulted in a significant RBCA lowering. The cell incubation with tirofiban and epifibatide issued in RBCA decrease. The similar results were obtained when RBCs were incubated with the fibrinogen inhibiting peptide. Although monafram, tirofiban, eptifibatide and the fibrinogen inhibiting peptide were related to fibrinogen function they didn't inhibit RBCA completely. Therefore, under moderate and low red blood cell aggregation the cell binding is probably related to nonspecific mode. It seems evident that the specific and nonspecific modes of red blood cell aggregate formation could co-exist. Additional theoretical and experimental investigations in this area are needed.
Heparinized blood better preserves cellular energy charge and vascular functions of intraoperatively stored saphenous vein grafts in comparison to isotonic sodium-chloride-solution
Authors: Wilbring Manuel , Ebner Annette , Schoenemann Katrin , Knaut Michael , Tugtekin Sems Malte , Zatschler Birgit , Waldow Thomas , Alexiou Konstantin , Matschke Klaus , Deussen Andreas
Published: December 2019

OBJECTIVE: Several studies have addressed the optimal storage conditions for vascular grafts during surgery. The results remain contradictionary. This may be attributed to the fact, that the various vascular beds have a different sensitivity to storage. We analyzed the impact of storage in isotonic saline solution (NaCl) or heparinized blood the vascular functions of human saphenous vein grafts. Special care was taken to choose storage conditions which are relevant for intraoperative storage of a saphenous vein graft in a setting of coronary artery bypass grafting with vein and internal mammary artery as grafts. METHODS: Intraoperatively isolated V. saphena-segments (n = 36) were stored in NaCl or heparinized blood for approximately 30 minutes at room temperature. Subsequently, the segments were examined in a Mulvany-myograph. Following preconstriction with norepinephrine, concentration-relaxation curves were assessed for bradykinin and sodium-nitroprusside to assess developed vessel-wall tension as well as endothelium- and smooth-muscle-cell dependent vasorelaxation. The availability of adenosintriphosphate (energy charge) was determined based on liquid chromatography measurements of nucleotide tissue levels. RESULTS: Mean storage time was 27.4 ± 2.4 min in NaCl- and 26.3 ± 2.7 min in blood-group, respectively. After this period, receptor-dependent and-independent maximum of developed vessel wall tension was significantly reduced in NaCl-group (p = 0.05 and p = 0.045, respectively). Furthermore, the energy charge was significantly (p = 0.046) better preserved after blood storage (74 ± 1%) in comparison to NaCl-group (68 ± 2%). Endothelium-induced vasodilatation in response to bradykinin reached only 12.3 ± 2.5% in NaCl-group, but 19.3 ± 5.2% in blood-group (p = 0.033). Alike, EC50-concentration of bradykinin for half-maximal relaxation was significantly lower in blood- than in NaCl-group (log EC50 −7.08 ± 0.3 and −5.91 ± 0.4; respectively; p = 0.046). Endothelium-independent smooth muscle relaxation in response to sodium-nitroprusside was not different between both groups. CONCLUSION: Heparinized blood better preserves vascular contractile and endothelial functions of the saphenous vein graft. Storage in NaCl rapidly compromises vascular functions and impaires cellular energy. NaCl should no longer be recommended for intraoperative storage of harvested V. saphena grafts.
Influence of fibre diameter and orientation of electrospun copolyetheresterurethanes on smooth muscle and endothelial cell behaviour
Authors: Rüder Constantin , Sauter Tilman , Kratz Karl , Haase Tobias , Peter Jan , Jung Friedrich , Lendlein Andreas , Zohlnhöfer Dietlind
Published: December 2019

Polymers exhibiting cell-selective effects represent an extensive research field with high relevance for biomedical applications e.g. in the cardiovascular field supporting re-endothelialization while suppressing smooth muscle cell overgrowth. Such an endothelial cell-selective effect could be recently demonstrated for a copolyetheresterurethane (PDC) containing biodegradable poly(p-dioxanone) and poly(ε-caprolactone) segments, which selectively enhanced the adhesion of human umbilical vein endothelial cells (HUVEC) while suppressing the attachment of smooth muscle cells (SMC). In this study we investigated the influence of the fibre orientation (random and aligned) and fibre diameter (2 μm and 500 nm) of electrospun PDC scaffolds on the adhesion, proliferation and apoptosis of HUVEC and SMC. Adhesion, viability and proliferation of HUVEC was diminished when the fibre diameter was reduced to a submicron scale, while the orientation of the microfibres did only slightly influence the cellular behaviour. In contrast, a submicron fibre diameter improved SMC viability. In conclusion, PDC scaffolds with micron-sized single fibres could be promising candidate materials for cell-selective stent coatings.
Cardiometabolic and traditional cardiovascular risk factors and their potential impact on macrovascular and microvascular function: Preliminary data
Authors: Gayda Mathieu , Juneau Martin , Tardif Jean-Claude , Harel François , Levesque Sylvie , Nigam Anil
Published: December 2019

We studied the relationship between cardiovascular status (CV) and risk factor numbers, macrovascular (flow mediated dilatation: FMD) and microvascular function using near-infrared spectroscopy (NIRS) in adults with different CV status. Seventy adults with different CV status (27 controls, 18 metabolic syndrome (MetS) and 25 coronary heart disease (CHD) patients) underwent a 5-min forearm arterial occlusion in supine position. High-resolution ultrasound examination of the brachial artery was performed during 1 minute at rest and 45 to 120 seconds after cuff release. Oxy, de-oxy and total hemoglobin signals (O2Hb, HHb and tHb) were measured continuously with NIRS on brachio-radialis muscle. FMD was reduced in CHD patients (P < 0.05) compared to controls. Max. amplitude of O2Hb and Hmax of tHb were reduced (P < 0.05) in MetS patients vs. controls. Post-deflation area under the curve (A.U.C) of O2Hb was lower in CHD (P < 0.01) patients vs. controls and MetS patients. Independent predictors of microvascular function (A.U.C of O2Hb) were abdominal obesity and LDL-cholesterol whereas macrovascular function (FMD) was predicted by CV status. Only A.U.C of O2Hb related to CV risk numbers whereas FMD was not. Although macro and microvascular function were impaired in MetS and CHD patients, microvascular function was more strongly related to CV risk factors.
Erythrocyte deformability and nitric oxide mobilization under pannexin-1 and PKC dependence
Authors: Silva-Herdade A.S. , Freitas T. , Almeida J. Pedro , Saldanha C.
Published: December 2019

The erythrocyte adenosine triphosphate (ATP) is utilised for protein phosphorylation and exported through the pannexin 1 hemichannel (Px1) in the microcirculation. The physiological stimuli for ATP release are dependent of blood shear rate level and of the tissue oxygen content. The deoxygenated and oxygenated states of haemoglobin are respectively bound and unbound to N terminal domain of the protein band 3 of the erythrocyte membrane in dependence of its degree of phosphorylation. The protein tyrosine kinase (PTK) and protein tyrosine phosphatase (PTP) contribute to the phosphorylation degree of band 3 and are modulated by protein kinase C (PKC). Chelerythrine (Che) is a competitive inhibitor of ATP for PKC and a negative modulator of erythrocyte deformability. The aim of this study was to assess the mobilization of nitric oxide (NO) in erythrocyte in absence and presence of Che and Px1 inhibitor (carbenoxolone). Erythrocyte deformability was evaluated in presence of carbenoxolone (Carb). Regarding the effects observed in the erythrocyte by presence of Che or Carb, the values of efflux of NO and the concentration of nitrosogluthatione are similar and with no changes in relation to their absence. Px1inhibition by Carb 10 μM ameliorates the erythrocyte deformability at a shear force of 0.6 and 1.2 Pa. The PKC inhibitor shows similar effects to the Carb on the mobilization of nitric oxide in erythrocyte. The blockage of ATP release by Carb from erythrocytes suggests a possible benefit to develop in ischemia reperfusion or in inflammatory response where will be needed to rescue the excess of NO present and ameliorate the red blood cell deformability at low shear rates.
Subclinical hypothyroidism and cardiovascular risk
Authors: Vayá Amparo , Giménez Cristina , Sarnago Ana , Alba Amparo , Rubio Olaya , Hernández-Mijares Antonio , Cámara Rosa
Published: December 2019

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.
Heart valve stenosis in laser spotlights: Insights into a complex disease
Authors: Büttner Petra , Galli Roberta , Jannasch Anett , Schnabel Christian , Waldow Thomas , Koch Edmund
Published: December 2019

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.
Genetic engineering of mesenchymal stem cells by non-viral gene delivery
Authors: Wang Weiwei , Xu Xun , Li Zhengdong , Lendlein Andreas , Ma Nan
Published: December 2019

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.
Expression pattern analysis and activity determination of matrix metalloproteinase derived from human macrophage subsets
Authors: Roch Toralf , Akymenko Oksana , Krüger Anne , Jung Friedrich , Ma Nan , Lendlein Andreas
Published: December 2019

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.
Radiographic contrast media alterate the localization of actin/band4.9 in the membrane cytoskeleton of human erythrocytes
Authors: Franke R.P. , Scharnweber T. , Fuhrmann R. , Mrowietz C. , Wenzel F. , Krüger A. , Jung F.
Published: December 2019

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.
A strategy for cell-based multiplex diagnostics of Myasthenia gravis and autoimmune encephalitis by modifying the subcellular localization of cell membrane autoantigens
Authors: George S. , Georgi M. , Roggenbuck D. , Conrad K. , Küpper J.-H.
Published: December 2019

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.
Influence of local hemostatic and antiplatelet agents on the incidence of bleeding complications in carotid endarterectomies
Authors: Weinrich M. , Schindler P. , Kundt G. , Klar E. , Bünger C.M.
Published: December 2019

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.
The role of VTIQ as a new tissue strain analytics measurement technique in testicular lesions
Authors: Trottmann M. , Marcon J. , D'Anastasi M. , Karl A. , Stief C.G. , Reiser M. , Clevert D.A.
Published: December 2019

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.
Increased red blood cell aggregation in patients with Gaucher disease is non-inflammatory
Authors: Adar T. , Ben-Ami R. , Elstein D. , Zimran P. , Berliner S. , Yedgar S. , Barshtein G.
Published: December 2019

Red blood cell (RBC) aggregation is enhanced in the presence of ongoing inflammation, because of plasma protein effects, especially fibrinogen. Large RBC aggregates, in addition to being a marker of systemic inflammation, may hinder tissue perfusion and oxygenation. Gaucher disease, the most common lysosomal storage disorder, evinces many of the hallmarks of chronic inflammation. Manifestations of Gaucher disease which may be related to microvascular occlusion include avascular necrosis (AVN), bone crisis, and pulmonary hypertension. This study aims to determine whether increased RBC aggregation in non-splenectomized patients with Gaucher disease is due to Gaucher-related inflammation. The Cell Flow Properties Analyzer (CFA) monitors blood under conditions of different shear stress by creating varying pressure gradients. Blood from non-splenectomized patients with Gaucher disease showed only a slight correlation between aggregation parameters and fibrinogen levels, whereas blood from non-splenectomized patients treated with enzyme replacement therapy (ERT) showed marked correlation between aggregation parameters and fibrinogen, as in the control group. These results underscore the hypothesis that RBC aggregation in Gaucher disease is increased by (at least) two mechanisms: a fibrinogen-mediated inflammatory process and another non-inflammatory process that may be induced by elevated glucocerebroside levels in the RBC and/or inhibited by elevated plasma cerebroside levels.
Microcirculation and perfusion with contrast enhanced ultrasound (CEUS) in Crohn's disease: First results with linear contrast harmonic imaging (CHI)
Authors: Schreyer A.G. , Finkenzeller T. , Gössmann H. , Daneschnejad M. , Müller-Wille R. , Schacherer D. , Zuber-Jerger I. , Strauch U. , Feuerbach S. , Jung E.M.
Published: December 2019

Aim: To evaluate a newly introduced high resolution linear transducer for vascularization and mural perfusion assessment using contrast harmonic imaging (CHI) with quantitative time intensity curve analysis (TIC) in patients with active Crohn's disease (CD).

Material and methods: We prospectively evaluated 14 consecutive patients (7 women, 7 males, age range 19–42 years, median 28 years) with histologically proven CD having an acute episode of the disease applying contrast enhanced MRI and high resolution ultrasound. For the ultrasound we used a newly introduced high resolution linear multi-frequency transducer (6–9 MHz, Logiq 9, GE). Ultrasound was performed by an experienced radiologist applying color coded Doppler sonography (CCDS), power Doppler (PD) and contrast enhanced CHI using the ‘true agent detection mode’. Additionally, 5 healthy volunteers were examined by ultrasound applying CCDS, PD and CHI. After the injection of 2.4 ml ultrasound contrast agent (SonoVue) the dynamic CHI cine sequences were recorded as digital raw data for 60 seconds. Therefore we were able to perform a quantitative perfusion analysis using TIC retrospectively. CCDS, PD and CHI with TIC were compared and analyzed.

Results: In all 14 patients MRI showed inflammatory changes in the terminal or pre-terminal ileum. Using PD and CCDS enlarged vessels surrounding the bowel wall were visualized in all patients. PD as well as CCDS diagnosed just in 9 of 14 patients augmented mural vessels. Having CHI with TIC increased mural contrast enhancement was diagnosed in all 14 patients. Patients with CD showed a maximum enhancement 36 s after injection with 9 dB (range 5.9–13.2 dB), while healthy volunteers reached the maximum level of 2.8 dB (range 2–3.8 dB) after 23 s (p<0.05).

Conclusion: Using high resolution linear transducer mural perfusion changes in active Crohn's disease can be appreciated applying CHI with TIC. This technique could be an effective dynamic imaging modality for diagnosis and especially follow-up examination to monitor treatment in CD.