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Andreas Deussen

Publications and source records attributed to Andreas Deussen.

15 recordsLinked to original sources
PubMed2007

The effects of lipid infusion on myocardial function and bioenergetics in l-bupivacaine toxicity in the isolated rat heart.

BACKGROUND: It is unclear whether improved metabolism or a "lipid sink" effect of lipid infusion is responsible for the positive effects in local anesthetic-induced myocardial depression. METHODS: We used an isolated rat heart, constant-pressure perfused, nonrecirculating Langendorff preparation and exposed hearts to 5 mug/mL l-bupivacaine and 9 microL/mL lipid emulsion. Hearts were freeze-clamped and energy was charge measured by HPLC. In a second experiment the effects of pacing hearts was evaluated. The effects of lipid addition on local anesthetic concentrations in Krebs-Henseleit buffer and human plasma were examined by using a mass spectrometer. RESULTS: With spontaneously beating hearts l-bupivacaine led to a significant decrease in heart rate (to 74% +/- 7% of baseline), +dP/dt (69% +/- 7%), systolic pressure (78% +/- 6%), coronary flow (61% +/- 8%), and to an increase in PR (177% +/- 52%) and QRS intervals (166% +/- 36%). Lipid infusion exerted a positive inotropic effect, significantly augmenting +dP/dt and systolic pressure back to 94% +/- 11% and 102% +/- 16% of baseline in l-bupivacaine-treated hearts. Heart rate, coronary flow, PR, and QRS intervals remained unchanged after lipid intervention. Lipid infusion in paced hearts had a significant effect on +dP/dt, systolic pressure, and Mvo2. Neither l-bupivacaine nor lipids had an effect on energy charge. A lipid concentration of 500 muL/mL plasma was necessary to effect changes in the plasma concentration of local anesthetics. CONCLUSION: Lipid application in l-bupivacaine-induced cardiac depression had a significant positive inotropic effect, which we would attribute to a direct inotropic effect. However, in an isolated heart model, indirect, local anesthetic plasma-binding effect of lipids cannot be excluded.

Animals↗
PubMed2006

Vardenafil increases coronary flow response to hypercapnic acidosis in isolated guinea pig heart.

The hypothesis was tested that vardenafil, a PDE5 inhibitor, specifically enhances coronary vasodilation during acidosis. In isolated constant pressure perfused guinea pig hearts, infusion of vardenafil (<or= 1 microM) increased coronary flow concentration-dependently 34 % above baseline. In parallel, cGMP release increased (0.44 +/- 0.094 vs. 0.14 +/- 0.017 pmol/min x g at 0.5 microM vardenafil vs. baseline). Flow increases occurred in the absence of changes in heart function (LVP, heart rate, dP/dt(max), heart rate - pressure product). Infusion of the NO synthase blocker L-NMMA (100 microM) caused a rightward shift of the dose-response curve of vardenafil. To test whether vardenafil treatment may enhance metabolic coronary vasodilation, arterial pCO(2) was raised from 38 to 61 mmHg, which resulted in a steady state flow increase of 18.8 +/- 4.5%. Infusion of vardenafil, given at a threshold flow enhancing concentration, doubled the coronary flow response during hypercapnic acidosis to 38.4 +/- 4.2 % (p=0.004). This flow amplification during acidosis was not shared by the K(ATP) channel opener cromakalim, indicating a specific effect of vardenafil on flow control during myocardial acidosis. We conclude that vardenafil specifically relaxes coronary resistance vessels through NO/cGMP-dependent pathways and increases the coronary flow response toward hypercapnic acidosis. This finding further supports the importance of the NO-cGMP axis in mediation of this flow response.

Acidosis↗
PubMed2006

Adenosine stimulates connexin 43 expression and gap junctional communication in pituitary folliculostellate cells.

Adenosine is known to stimulate interleukin (IL)-6 and vascular endothelial growth factor (VEGF) secretion from pituitary TtT/GF folliculostellate [corrected] (FS) cells indicating that it is an important paracrine regulator of anterior pituitary function. This study demonstrates that rodent anterior pituitary cell lines produce extracellular adenosine that is able to increase intercellular gap junction communication in FS cells. Ecto-5'-nucleotidase (CD73), the enzyme that generates adenosine from AMP, was demonstrated by immunocytochemistry in approximately 20% of anterior pituitary cells, and some of these cells colocalized with prolactin and growth hormone. CD73 mRNA and protein were detected in GH3 and MMQ (somatotroph-lactotroph lineages) and TtT/GF cells, and enzyme activity was demonstrated by the conversion of exogenously added fluorescent ethenoAMP to ethenoadenosine. Adenosine production, as measured by HPLC, was detected in GH3 (1 microM/h) and MMQ (3 microM/h) but not in TtT/GF cells. Adenosine (EC50: 0.5 microM) and NECA (universal adenosine receptor agonist; EC50 0.1 microM) stimulated connexin 43 (Cx43) mRNA and protein expression within 1-2 h in TtT/GF cells. Adenosine and NECA also stimulated gap junctional intercellular communication (as assessed by transmission of Alexa Fluor 488) by 6- to 8-fold in comparison with untreated TtT/GF cells. In cocultures of MMQ and TtT/GF cells, Cx43 expression in TtT/GF cells increased in proportion to the number of MMQ cells plated out. These data suggest that adenosine, formed locally in the anterior pituitary gland can stimulate gap junction communication in FS cells.

5'-Nucleotidase↗
PubMed2005

Adenosine concentration in the porcine coronary artery wall and A2A receptor involvement in hypoxia-induced vasodilatation.

We tested whether hypoxia-induced coronary artery dilatation could be mediated by an increase in adenosine concentration within the coronary artery wall or by an increase in adenosine sensitivity. Porcine left anterior descendent coronary arteries, precontracted with prostaglandin F(2alpha) (10(-5) M), were mounted in a pressure myograph and microdialysis catheters were inserted into the tunica media. Dialysate adenosine concentrations were analysed by HPLC. Glucose, lactate and pyruvate were measured by an automated spectrophotometric kinetic enzymatic analyser. The exchange fraction of [(14)C]adenosine over the microdialysis membrane increased from 0.32 +/- 0.02 to 0.46 +/- 0.02 (n = 4, P < 0.01) during the study period. At baseline, interstitial adenosine was in the region of 10 nM which is significantly less than previously found myocardial concentrations. Hypoxia (P(O(2)) 30 mmHg for 60 min, n = 5) increased coronary diameters by 20.0 +/- 2.6% (versus continuous oxygenation -3.1 +/- 2.4%, n = 6, P < 0.001) but interstitial adenosine concentration fell. Blockade of adenosine deaminase (with erythro-9-(2-hydroxy-3-nonyl-)-adenine, 5 microM), adenosine kinase (with iodotubericidine, 10 microM) and adenosine transport (with n-nitrobenzylthioinosine, 1 microM) increased interstitial adenosine but the increase was unrelated to hypoxia or diameter. A coronary dilatation similar to that during hypoxia could be obtained with 30 microM of adenosine in the organ bath and the resulting interstitial adenosine concentrations (n = 5) were 20 times higher than the adenosine concentration measured during hypoxia. Adenosine concentration-response experiments showed vasodilatation to be more pronounced during hypoxia (n = 9) than during normoxia (n = 9, P < 0.001) and the A(2A) receptor antagonist ZM241385 (20 nM, n = 5), attenuated hypoxia-induced vasodilatation while the selective A(2B) receptor antagonist MRS1754 (20 nM, n = 4), had no effect. The lactate/pyruvate ratio was significantly increased in hypoxic arteries but did not correlate with adenosine concentration. We conclude that hypoxia-induced coronary artery dilatation is not mediated by increased adenosine produced within the artery wall but might be facilitated by increased adenosine sensitivity at the A(2A) receptor level.

Acetamides↗
PubMed2005

Human osteoblast precursors produce extracellular adenosine, which modulates their secretion of IL-6 and osteoprotegerin.

UNLABELLED: We showed that human osteoprogenitor cells produced adenosine and expressed ecto-5'-nucleotidase and all four adenosine receptor subtypes. Adenosine stimulated IL-6 but inhibited osteoprotegerin secretion, suggesting that adenosine is a newly described regulator of progenitor cell function. INTRODUCTION: Maintaining skeletal homeostasis relies on there being a balance between bone formation and resorption; an imbalance between these processes can lead to diseases such as osteoporosis and rheumatoid arthritis. Recent reports showed that locally produced ATP, acting through P2 receptors, has pronounced effects on bone formation. However, ATP can be enzymatically cleaved to adenosine that has little or no activity at P2 receptors but mediates its action through the P1 family of receptors. We studied whether adenosine may also have an important role in controlling bone cell differentiation and function. MATERIALS AND METHODS: Extracellular adenosine levels were analyzed by high-performance liquid chromatography in HCC1 and bone marrow stromal (BMS) cells. Ecto-5'-nucleotidase (CD73) expression and activity was determined by RT-PCR, immunocytochemistry, and the cleavage of etheno-AMP to ethenoadenosine. Adenosine receptor expression and activity were determined by RT-PCR and cAMP measurements. The effects of adenosine receptor agonists on IL-6, osteoprotegerin (OPG), and RANKL expression were determined by ELISA and QRT-PCR. RESULTS: HCC1 and BMS cells produce adenosine and express CD73 and all four adenosine receptor subtypes. The A2b receptor was shown to be functionally dominant in HCC1 cells, as determined by cAMP production and in its stimulation of IL-6 secretion. Adenosine receptor agonism also inhibited OPG secretion and OPG but not RANKL mRNA expression. CONCLUSIONS: Our findings show that HCC1 and primary BMS cells produce adenosine, express CD73 and all four adenosine receptor subtypes. In HCC1 cells, adenosine has a potent stimulatory action on IL-6 secretion but an inhibitory action on OPG expression. These data show for the first time that adenosine may be an important regulator of progenitor cell differentiation and hence an important local contributor to the regulation of bone formation and resorption.

5'-Nucleotidase↗
PubMed2005

Modulation of ecto-5'-nucleotidase by phospholipids in human umbilical vein endothelial cells (HUVEC).

Ecto-5'-nucleotidase, the major enzyme controlling extracellular adenosine production, can be activated by phospholipids, e.g. lysophosphatidylcholine (LPC). This study examined the structural requirements of phospholipids to evoke this enzyme activation and figured out two new activators of ecto-5'-nucleotidase: platelet activating factor (PAF) and sphingosylphosphorylcholine (SPC). Potential signal transduction pathways including an involvement of protein kinase C and PAF-receptor were evaluated on the model of human umbilical vein endothelial cells (HUVEC). Cells were pre-incubated with 10 microM of various phospholipids including lysophosphatidylcholine, beta-arachidonyl-gamma-palmityl-alpha-phosphatidylcholine, beta,gamma-dipalmityl-alpha-phosphatidyl-choline, beta,gamma-dipalmityl-alpha-phosphatidylethanolamine, beta,gamma-dipalmityl-alpha-phosphatidylserine, gamma-acyl-beta-lyso-alpha-phosphatidylethanolamine, beta-acetyl-gamma-O-hexadecyl-alpha-phosphatidylcholine (platelet activating factor), lysophosphatidylic acid, sphingosine-1-phosphate and sphingosylphosphorylcholine. In the cell supernatant the extracellular dephosphorylation rate of the fluorescent AMP-analogue 1,N6-etheno-5'AMP to 1,N6-etheno-adenosine was measured by HPLC. Out of these ten structurally related phospholipids only lysophosphatidylcholine, sphingosylphosphatidylcholine and platelet activating factor dose-dependently increased the activity of ecto-5'-nucleotidase. Pharmacological blocking experiments revealed that neither the activation of PAF-receptor nor of protein kinase C were important for mediating the activation of ecto-5'-nucleotidase. Thus, using information on the known molecular structures of tested phospholipids, a phosphatidylcholine residue in alpha-position and a short chain length fatty acid esterified in beta-position seem essential for activation of ecto-5'-nucleotidase by glycerophospholipids. Since all tested phospholipids have similar fatty acid chain lengths and residues in alpha-position, they should act similarly on membrane fluidity. It is concluded that the observed effects are not based on changes in membrane fluidity by the added phospholipids, but rather involve a yet to be determined phospholipid-receptor.

5'-Nucleotidase↗
PubMed2005

External compression increases forearm perfusion.

Application of compression stockings to the lower extremities is a widely used therapeutic intervention to improve venous return, but there is little information about the effects of compression on local arterial perfusion. Therefore, we tested the hypothesis that a positive external pressure increases forearm perfusion. The relation of increasing external pressure induced by standardized compression to the arterial inflow and arterial flow reserve of the forearm was critically evaluated in a group of healthy young men (n = 9). Flow was measured with venous occlusion plethysmography after a 10-min application of six different stockings with compression pressure increasing from 13 to 23 mmHg. During compression, the arterial inflow increased significantly from 3.7 +/- 0.85 to 8.8 +/- 2.01 ml.min(-1).100 ml tissue(-1) (P < 0.001) and the arterial flow reserve increased from 17.7 +/- 4.7 to 28.3 +/- 7.0 ml.min(-1).100 ml tissue(-1). The flow increase was persistent after 3 h of constant application of external pressure and also during simultaneous low-intensity hand grip. Similar results obtained with occlusion plethysmography were seen with MRI. During the interventions, forearm temperature was unchanged, and the volunteers reported no discomfort. In conclusion, 1) arterial perfusion of the human forearm increases more than twofold during application of external compression over a pressure range of 13-23 mmHg, and 2) the result is interpreted as an autoregulatory response following the decrease of the vascular transmural pressure gradient.

Adult↗
PubMed2005

Effects of homocysteine on vascular and tissue adenosine: a stake in homocysteine pathogenicity?

Homocysteine may have deleterious effects on the cardiovascular system. It has been hypothesized that these effects may be brought about by a decrease in the adenosine concentration via the S-adenosylhomocysteine hydrolase reaction. A requirement for this causal relationship is proof of a reduction in vascular adenosine concentration during conditions of elevated homocysteine concentrations. In the present communication we summarize published data obtained during systematic variation of the arterial homocysteine concentration. Most of the results reported show that an increase in homocysteine concentration to 100 microM is associated with a 20-50% decrease in vascular adenosine concentration and an increase in tissue S-adenosylhomocysteine level. Homocysteine effects on the adenosine concentration seem to be more pronounced under conditions of impaired oxygenation. Further experiments, in particular on organs and tissue that release high amounts of homocysteine, i.e., the liver, are warranted to study the potential effects of homocysteine on vascular and tissue adenosine concentrations and consequent effects on organ function. The evidence obtained may be relevant for future assessment of risk indicators in conjunction with homocysteine pathogenicity, which might potentially be extended to measurements of adenosine or S-adenosylhomocysteine levels.

Adenosine↗
PubMed2004

Intact nitric oxide production is obligatory for the sustained flow response during hypercapnic acidosis in guinea pig heart.

OBJECTIVE: The mechanisms underlying hypercapnic coronary dilation remain unsettled. This study tests the hypothesis that flow dependent NO production is obligatory for the hypercapnic flow response. METHODS/RESULTS: In isolated, constant pressure (CP) perfused guinea pig hearts a step change of arterial pCO(2) from 38.6 to 61.4 mm Hg induced a bi-phasic flow response with an early transient (maximum 60 s) and a consecutive persisting flow rise (121.6+/-6.6 (S.D.) % after 10 min). In contrast, when perfused with constant flow (CF), perfusion pressure only transiently (2 min) fell by 7.4+/-4.8 % following the step change of arterial pCO(2). In CP perfused hearts L-NAME (100 micromol/l) specifically abolished the delayed flow rise during hypercapnic acidosis (102.37+/-2.9% after 10 min), whereas the inhibitor had no effect on perfusion pressure response in CF perfused hearts. Under CP perfusion arterial hypercapnia resulted in a transient rise of coronary cGMP release (from 0.69+/-0.35 to 1.12+/-0.68 pmol/ml), which was abolished after L-NAME. Surprisingly, the K(+)ATP channel blocker glibenclamide did not have any significant effect on the hypercapnic flow response but largely blunted reactive hyperemia after a 20 s flow stop. CONCLUSIONS: The delayed steady state hypercapnic flow response in guinea pig heart requires intact NO production. The absence of a persisting decrease in coronary resistance under CF perfusion points to an important role of shear stress dependent NO production.

Acidosis↗
PubMed2004

Na(+)-D-glucose cotransporter in muscle capillaries increases glucose permeability.

By immunohistochemistry, we demonstrated the localization of the Na(+)-D-glucose cotransporter SGLT1 in capillaries of rat heart and skeletal muscle, but not in capillaries of small intestine and submandibular gland. mRNA of SGLT1 was identified in skeletal muscle and primary cultured coronary endothelial cells. The functional relevance of SGLT1 for glucose transport across capillary walls in muscle was tested by measuring the extraction of D-glucose from the perfusate during non-recirculating perfusion of isolated rat hindlimbs. In this model, D-glucose extraction from the perfusate is increased by insulin which accelerates D-glucose uptake into myocytes by increasing the concentration of glucose transporter GLUT4 in the plasma membrane. The insulin-induced increase of D-glucose extraction from the perfusate was abolished after blocking SGLT1 with the specific inhibitor phlorizin. The data show that SGLT1 in capillaries of skeletal muscle is required for the action of insulin on D-glucose supply of myocytes.

Animals↗
PubMed2003

Modulation of adenine nucleotide concentrations in human plasma by erythrocytes and endothelial cells.

The regulation of plasma concentrations of adenine nucleotides is unsettled. We tested the possibility of extracellular adenosine triphosphate (ATP) production from adenosine diphosphate (ADP) at physiological low concentrations by erythrocytes and endothelial cells. Filtered erythrocytes and human umbilical vein endothelial cells (HUVEC) were incubated for 15 to 120 s with ADP (10 microM), supplemented with 3H-ADP (2.85 nM) or 14C-ADP (54.6 nM). Enzymatic conversion of ADP to ATP was detected by recovery of the radioactive label in the ATP fraction. ATP was measured in the supernatant using high performance liquid chromatography (HPLC) separation, scintillation techniques, and luminometry. Using etheno (epsilon)-labeled ADP (10 microM), the extracellular localization of the conversion was further corroborated. Following ADP application in plasma, no radioactivity was detected in the ATP fraction. However, in erythrocyte suspensions, 12.9% and 9.7% of the label were recovered in the ATP fraction after application of 3H- and 14C-ADP, respectively. Between 15 and 120 s after 3H-ADP application, the 3H-ATP fraction was found to be stable at around 10%. For the range of ADP concentrations studied (10-40 microM), no saturation of ATP production was achieved. The extracellular localization of conversion was supported by the recovery of the epsilon -label in the epsilon -ATP fraction. In contrast, on HUVEC a conversion of epsilon -ADP to epsilon -ATP was not observed. In conclusion, on erythrocytes there is rapid enzymatic conversion of extracellular ADP to ATP which may play a significant role in adjusting adenine nucleotide concentrations in human plasma. In endothelial cells, extracellular conversion of ADP to ATP is of quantitatively minor importance, if it contributes at all.

Adenine Nucleotides↗
PubMed2003

Liquid chromatography method for the analysis of adenosine compounds.

A newly available chromatography column packing material that employs hybrid particle technology was used to improve the analysis of adenosine compounds. Using a TBAS buffer/acetonitrile gradient this material permits separation of etheno-adenosine compounds in less than 4 min with excellent resolution and sensitivity (50 fmol). Variability of compound quantification is small (coefficients of variation 0.23+/-0.14% for 50 pmol and 1.70+/-0.53% for 0.5 pmol). The new method is well suited for the analysis of adenosine compounds in small biological samples and permits a high sample throughput in autosampler setups with high precision and reproducibility.

Adenosine↗
PubMed2003

Direct biochemical evidence for eNOS stimulation by bradykinin in the human forearm vasculature.

OBJECTIVE: Although it has been shown recently that acetylcholine (ACh)-induced vasodilation of forearm resistance vessels is predominantly mediated by nitric oxide, direct biochemical evidence for eNOS stimulation by bradykinin (BK) in the human arterial circulation is still lacking. Therefore, the present study was designed to test the hypothesis that in the human forearm vasculature eNOS stimulation significantly contributes to BK-induced vasodilation. METHODS: BK was infused in the presence and absence of the NOS inhibitor L-NMMA (8 micromol/min) into the brachial artery of 16 healthy volunteers and the effects compared to muscarinergic eNOS stimulation following acetylcholine infusion. Forearm blood flow (FBF) was measured by venous occlusion plethysmography, and plasma nitrite (NO(2)(-)), which represents a sensitive and specific marker of regional eNOS activity, was determined in the antecubital vein and brachial artery by flow injection analysis. Nitric oxide production was calculated as product of the veno-arterial difference of NO(2)(-) concentration times FBF. RESULTS: Kininergic (BK: 20, 60, 200 ng/min) as well as muscarinergic (ACh: 1, 3, 10 microg/min) stimulation resulted in a dose-dependent increase in FBF and NO(2)(-) in each individual. The relationship between FBF and NO production upon BK infusion was comparable to that obtained with ACh (r = 0.98; n = 64, p < 0.01). Moreover, NOS inhibition reduced both flow responses and NO production (BK: 54 and 75 %; ACh: 57 and 72 %) to a similar extent. CONCLUSIONS: These data provide direct biochemical evidence for the involvement of eNOS in bradykinin-induced vasodilation of forearm resistance vessels in humans.

Acetylcholine↗
PubMed2003

Tracer adenosine: a novel myocardial flow marker.

UNLABELLED: Local myocardial blood flow measurements are of great importance in experimental and clinical settings. However, a lack of ideal markers is evident. Adenosine is suggested to be a potential candidate because of its high uptake and rapid intracellular sequestration. We specifically tested the hypothesis that the local deposition density of labeled adenosine within the heart reflects local myocardial blood flow. METHODS: Tracer microspheres, the recognized standard for local blood flow measurements, were injected and compared with simultaneously injected labeled adenosine ((3)H/(14)C) in tracer concentration into the left atrium of anesthetized Beagle dogs (n = 7). Myocardial deposition densities were assessed through beta-scintillation and gamma-counting measurements in samples (100-128 per heart) of an average wet mass of 487 +/- 54 mg. To challenge local myocardial blood flow distribution, alprostadil was infused into the left circumflex artery in 3 experiments. In 2 other experiments, erythro-9-hydroxy-nonyl-adenine (EHNA) was infused to inhibit degradation of injected adenosine to inosine. RESULTS: Tracer adenosine and microspheres did not exert significant local or systemic hemodynamic effects. Both were almost completely extracted from blood within 2 min and locally retained in the tissue. Deposition densities of tracer microspheres and labeled adenosine correlated closely in each experiment, independently of the respective protocol (control, EHNA, or alprostadil), over a wide range of local myocardial blood flows (0.23-12.9 mL min(-1) g(-1)). The mean correlation coefficient (n = 293) was r = 0.93 (r(2) = 0.86; P < 0.0001), indicating that the deposition density of (3)H-adenosine could explain local blood flow as measured with the tracer microsphere technique with 86% probability. CONCLUSION: Adenosine appears to be a reliable marker of local blood flow in dog myocardium.

Adenine↗