Gabriele Nöldge-Schomburg

Impact of inspired substance concentrations on the results of breath analysis in mechanically ventilated patients

Abstract A well-defined relationship has to exist between substance concentrations in blood and in breath if blood-borne volatile organic compounds (VOCs) are to be used as breath markers of disease or health. In this study, the impact of inspired substances on this relationship was investigated systematically. VOCs were determined in inspired and expired air and in arterial and mixed venous blood of 46 mechanically ventilated patients by means of SPME, GC/MS. Mean inspired concentrations were 25% of expired concentrations for pentane, 7.5% for acetone, 0.7% for isoprene and 0.4% for isoflurane. Only if inspired concentrations were <5% did substance disappearance rates from blood and exhalation rates correlate well. Exhaled substance concentrations depended on venous and inspired concentrations. Patients with sepsis had higher n-pentane and lower acetone concentrations in mixed venous blood than patients without sepsis (2.27 (0.37–8.70) versus 0.65 (0.33–1.48) nmol L−1 and 69 (22–99) versus 18 (6.7–56) µmol L−1). n-Pentane and acetone concentrations in breath showed no differences between the patient groups, regardless whether or not expired concentrations were corrected for inspired concentrations. In mechanically ventilated patients, concentration profiles of volatile substances in breath may considerably deviate from profiles in blood depending on the relative amount of inspired concentrations. A simple correction for inspired substance concentrations was not possible. Hence, substances having inspired concentrations >5% of expired concentrations should not be used as breath markers in these patients without knowledge of concentrations in blood and breath.

The effects of desflurane on splanchnic hemodynamics and oxygenation in the anesthetized pig

This study was designed to investigate the effects of desflurane on systemic and splanchnic hemodynamics, O2 delivery and O2 uptake, tissue oxygenation (as monitored by surface PO2 electrodes), and hepatic oxygen-dependent intermediary metabolism (hepatic lactate uptake, intestinal lactate production, ketone-body ratio) in the pig. We studied 11 anesthetized (i.e., ketamine, flunitrazepam, vecuronium) and ventilated domestic pigs (17-23 kg). After instrumentation, desflurane was administered randomly at 0.5 minimum alveolar anesthetic concentration (MAC) (4.2 vol %) and 1.0 MAC (8.3 vol %). Desflurane caused dose-dependent decreases in heart rate, mean arterial blood pressure, and cardiac output. Hepatic arterial blood flow was not affected at 0.5 MAC but decreased at 1.0 MAC. In contrast, portal and superior mesenteric arterial blood flow decreased at 0.5 MAC but did not show any further significant decrease at 1.0 MAC. Total hepatic blood flow decreased dose-dependently. Although O2 deliveries of whole body, liver, and small intestine were markedly reduced at both concentrations, respective O2 uptakes did not change significantly. The decreases in O2 deliveries were reflected by moderate disturbances in hepatic and small intestinal surface PO2. No evidence for severe tissue hypoxia could be detected. Desflurane had no adverse effects on hepatic and small intestinal metabolic function. These data indicate that hepatic and small intestinal O2 reserve capacity is impaired by desflurane. (Anesth Analg 1997;84:271-7)

Different effects of early endotoxaemia on hepatic and small intestinal oxygenation in pigs

ObjectiveStudy on simultaneous O2 supply/uptake relationships in liver and gut during endotoxaemia, to determine whether signs of dysoxia develop uniformly in the splanchnic region.DesignAnimal study to assess the early effects of endotoxaemia on oxygenation of both liver and small intestine.InterventionsEight anaesthetized pigs received a continuous portal venous infusion of lipopolysaccharide (0.5 μg·kg−1·h−1) for 6 h. Systemic, pulmonary and splanchnic haemodynamics as well as systemic and splanchnic O2 supply/uptake relationships were determined.ResultsThere was a multiphasic haemodynamic response pattern characterized by an early (within the 1st h) and a subsequent more prolonged phase (between the 2nd and 6th h) of decreases and recovery of hepatic arterial, portal venous and superior mesenteric arterial blood flows (electromagnetic flow probes) and splanchnic O2 deliveries. Unrelated to perfusion pressure and O2 delivery, there were early and sustained decreases in ileal mucosal surface partial pressure of oxygen (PO2) (multiwire PO2 electrode) and pH (tonometry). This was not reflected by ileal serosal surface PO2, O2 uptake and arteriomesenteric venous pH and partial pressure of carbon dioxide (PCO2) gradients. There was little evidence of concomitant hepatic dysoxia as evaluated by surface PO2.ConclusionsThe study demonstrates early and sustained regional (mucosa) intestinal hypoxia with little evidence of simultaneous hepatic dysoxia during initial endotoxaemia.

The neuronal guidance protein netrin-1 reduces alveolar inflammation in a porcine model of acute lung injury

IntroductionAcute lung injury (ALI) is an inflammatory disorder of pulmonary or extrapulmonary origin. We have previously demonstrated that netrin-1 dampens murine ALI, and in an attempt to advance this finding into future clinical practice we evaluated whether netrin-1 would reduce alveolar inflammation during porcine ALI.MethodsThis was a controlled in vivo experimental study in pigs. We induced ALI through lipoploysaccharide (LPS) infusion (50 μg/kg) for 2 hours. Following this, we exposed animals to either vehicle, intravenous netrin-1 (netrin-1 i.v.) or inhaled netrin-1 (netrin-1 inh.). Serum samples and bronchoalveolar lavage (BAL) were obtained to determine levels of tumor necrosis factor-α (TNF-α), interleukin (IL)-1β, interleukin-6 and interleukin-8 at baseline and 6 hours following treatment. Myeloperoxidase activity (MPO) and protein levels were determined in the BAL, and tissue samples were obtained for histological evaluation. Finally, animals were scanned with spiral CT.ResultsFollowing LPS infusion, animals developed acute pulmonary injury. Serum levels of TNF-α and IL-6 were significantly reduced in the netrin-1 i.v. group. BAL demonstrated significantly reduced cytokine levels 6 hours post-netrin-1 treatment (TNF-α: vehicle 633 ± 172 pg/ml, netrin-1 i.v. 84 ± 5 pg/ml, netrin-1 inh. 168 ± 74 pg/ml; both P < 0.05). MPO activity and protein content were significantly reduced in BAL samples from netrin-1-treated animals. Histological sections confirmed reduced inflammatory changes in the netrin-1-treated animals. Computed tomography corroborated reduced pulmonary damage in both netrin-1-treated groups.ConclusionsWe conclude that treatment with the endogenous anti-inflammatory protein netrin-1 reduces pulmonary inflammation during the initial stages of ALI and should be pursued as a future therapeutic option.

Toll-Like Receptor 2–Blocking Antibodies Promote Angiogenesis and Induce ERK1/2 and AKT Signaling via CXCR4 in Endothelial Cells

Objective—Toll-like receptor 2 (TLR2) inhibition by function blocking antibodies (ABs) is associated with enhanced preservation of endothelial cell function during vascular disease. In the present study, we investigated the capacity of TLR2-blocking ABs to modulate the angiogenic response of endothelial cells in vitro and in vivo. Approach and Results—Incubation of endothelial cells with mono- or polyclonal anti-TLR2 ABs resulted in increased tube formation, sprouting, and migration of endothelial cells compared with controls. In a mouse model of hindlimb ischemia, using TLR2-deficient or anti-TLR2 AB–treated wild-type mice resulted in increased new capillary formation and enhanced reperfusion. The effects of anti-TLR2 ABs were similar to those exerted by stromal cell–derived factor-1, and we show that anti-TLR2 ABs yet not TLR2 ligands lead to comparable activation of extracellular signal–regulated kinase1/2 and AKT but not p38 mitogen-activated protein kinase as activation of the CXCR4 canonical signal transduction pathways by stromal cell–derived factor-1. Immunoprecipitation of TLR2 revealed that anti-TLR2 ABs initiate an association of TLR2 with CXCR4 and mitogen-activated protein kinase activation. The proangiogenic properties of anti-TLR2 ABs were abolished by both G-protein inhibition and CXCR4 knockdown in endothelial cells. Conclusions—Our results provide evidence for a proangiogenic effect of TLR2-blocking ABs on endothelial cells in vitro and in vivo. They identify a novel molecular mechanism linking TLR2 to angiogenic processes that is independent from the activation of inflammatory cascades and further support the concept of a beneficial effect of TLR2 inhibition for endothelial cell function in vascular disease.

Extracorporeal cell therapy with granulocytes in a pig model of Gram-positive sepsis.

Objectives:Granulocyte transfusions have been used to treat immune cell dysfunction in sepsis. As granulocyte transfusions can trigger tissue injury via local effects of neutrophils, we hypothesized that extracorporeal treatment of plasma using granulocytes would prove beneficial while having less side effects. Design:Prospective controlled three-armed animal study. Setting:Research laboratory. Subjects:Twenty-one female immature pigs (7.5–12 kg, 7–9 weeks old). Interventions:Three groups of spontaneously breathing, sedated pigs (n = 7 each) received an intravenous lethal dose of live Staphylococcus aureus over 1 hour. Although group I had no specific treatment (control), group II and III were subsequently treated for 4 hours with an extracorporeal device containing either no cells (sham control, group II) or human cell line-derived granulocytic cells (group III). Survival time and physiologic, biochemical, and hematologic parameters were monitored for 7 days. Measurements and Main Results:All animals of group I died during the observation time (mean survival time: 70 hours). In group II, two of seven and in group III, six of seven animals survived the observation time (mean survival: 75 and 168 hours, respectively). Survival differences were significant between group I and III (p < 0.001) and between group II and III (p < 0.05) but not between group I and II (p = 0.43). Furthermore, group differences in bacterial blood concentrations, differential blood count, blood gases, lactate, and interleukins were observed. The extracorporeal cell treatment was well tolerated by the animals. Conclusions:Extracorporeal therapy with granulocytic cells significantly improved survival in a pig model of sepsis. Further studies with this approach are encouraged.

Extracorporeal cell therapy of septic shock patients with donor granulocytes: a pilot study

IntroductionNeutrophil granulocytes are the first defense line in bacterial infections. However, granulocytes are also responsible for severe local tissue impairment. In order to use donor granulocytes, but at the same time to avoid local side effects, we developed an extracorporeal immune support system. This first-in-man study investigated whether an extracorporeal plasma treatment with a granulocyte bioreactor is tolerable in patients with septic shock. A further intention was to find suitable efficacy end-points for subsequent controlled trials.MethodsThe trial was conducted as a prospective uncontrolled clinical phase I/II study with 28-day follow-up at three university hospital intensive care units. Ten consecutive patients (five men, five women, mean age 60.3 ± 13.9 standard deviation (SD) years) with septic shock with mean ICU entrance scores of Acute Physiology and Chronic Health Evaluation (APACHE) II of 29.9 ± 7.2 and of Simplified Acute Physiology Score (SAPS) II of 66.2 ± 19.5 were treated twice within 72 hours for a mean of 342 ± 64 minutes/treatment with an extracorporeal bioreactor containing 1.41 ± 0.43 × 10E10 granulocytes from healthy donors. On average, 9.8 ± 2.3 liters separated plasma were treated by the therapeutic donor cells. Patients were followed up for 28 days.ResultsTolerance and technical safety during treatment, single organ functions pre/post treatment, and hospital survival were monitored. The extracorporeal treatments were well tolerated. During the treatments, the bacterial endotoxin concentration showed significant reduction. Furthermore, noradrenaline dosage could be significantly reduced while mean arterial pressure was stable. Also, C-reactive protein, procalcitonin, and human leukocyte antigen DR (HLA-DR) showed significant improvement. Four patients died in the hospital on days 6, 9, 18 and 40. Six patients could be discharged.ConclusionsThe extracorporeal treatment with donor granulocytes appeared to be well tolerated and showed promising efficacy results, encouraging further studies.Trial registrationClinicalTrials.gov Identifier: NCT00818597

Design and implementation of a control system reflecting the level of analgesia during general anesthesia.

Abstract Introduction: Measuring and ensuring an adequate level of analgesia in patients are of increasing interest in the area of automated drug delivery during general anesthesia. Therefore, the aim of this investigation was to develop a control system that may reflect the intraoperative analgesia value. Our hypothesis was that a feedback controller could be applied in clinical practice safely and at an adequate quality of analgesia. The purpose of this study was to evaluate the practical feasibility of such a system in a clinical setting. Methods: The control system for the level of analgesia described in this paper relies on a parameter combination of heart rate variability (HRV), heart rate (HR), and blood pressure (mean arterial pressure, MAP), which serve as input variables for an expert system. For this fuzzy system, the experience of the participating anesthesiologists was translated into a set of fuzzy rules. In a pilot trial, the control system for automated titration of remifentanil, a short-acting opioid, was tested combined with a closed-loop propofol infusion system for hypnosis. Ten adult patients (4 women, 6 men), aged 22–52 years (median, 45 years; range, 29–49 years), with an American Society of Anesthesiologists physical status class I or II and who were scheduled for elective trauma surgery in a supine position were enrolled in this prospective trial. The precision of the system was calculated using internationally defined performance parameters. Results: There was no human intervention necessary during the computer-controlled administration of propofol and remifentanil, and operating conditions were satisfactory in all patients. All patients assessed the quality of anesthesia as “good” to “very good”. Median performance error, median absolute performance error, and wobble for HR and MAP during maintenance of anesthesia were -8.98 (5.32), 10.08 (4.17), and 2.68 (1.29) and -4.51 (12.73), 13.63 (2.27), and 3.90 (2.08) [mean (SD)], respectively. Conclusion: The control system, reflecting the level of analgesia during general anesthesia designed and evaluated in this study, allows for a clinically practical, nearly fully automated infusion of an opioid during medium-length surgical procedures with acceptable technical requirements and an adequate precision.

Awake Extracorporeal Membrane Oxygenation (ECMO) as Bridge to Recovery After Left Main Coronary Artery Occlusion: A Promising Concept of Haemodynamic Support in Cardiogenic Shock

Cardiogenic shock following acute myocardial infarction is associated with high mortality rate. Different management concepts including fluid management, inotropic support, intra aortic balloon counterpulsation (IABP) and extracorporeal membrane oxygenation (ECMO) mainly in mechanically ventilated patients have been used as cornerstones of management. However, success rates have been disappointing. Few reports suggested that ECMO when performed under circumvention of mechanical ventilation, may offer some survival benefits. We herein present our experience with the use of veno-arterial ECMO as bridge to recovery in an awake and spontaneously breathing patient after left main coronary artery occlusion complicated by cardiogenic shock.

Impact of Toll-Like Receptor 2 Deficiency on Survival and Neurological Function after Cardiac Arrest: A Murine Model of Cardiopulmonary Resuscitation

Background Cardiac arrest (CA) followed by cardiopulmonary resuscitation (CPR) is associated with poor survival rate and neurofunctional outcome. Toll-like receptor 2 (TLR2) plays an important role in conditions of sterile inflammation such as reperfusion injury. Recent data demonstrated beneficial effects of the administration of TLR2-blocking antibodies in ischemia/reperfusion injury. In this study we investigated the role of TLR2 for survival and neurofunctional outcome after CA/CPR in mice. Methods Female TLR2-deficient (TLR2-/-) and wild type (WT) mice were subjected to CA for eight min induced by intravenous injection of potassium chloride and CPR by external chest compression. Upon the beginning of CPR, n = 15 WT mice received 5 µg/g T2.5 TLR2 inhibiting antibody intravenously while n = 30 TLR2-/- and n = 31 WT controls were subjected to injection of normal saline. Survival and neurological outcome were evaluated during a 28-day follow up period. Basic neurological function, balance, coordination and overall motor function as well as spatial learning and memory were investigated, respectively. In a separate set of experiments, six mice per group were analysed for cytokine and corticosterone serum levels eight hours after CA/CPR. Results TLR2 deficiency and treatment with a TLR2 blocking antibody were associated with increased survival (77% and 80% vs. 51% of WT control; both P < 0.05). Neurofunctional performance was less compromised in TLR2-/- and antibody treated mice. Compared to WT and antibody treated mice, TLR2-/- mice exhibited reduced IL-6 (both P < 0.05) but not IL-1β levels and increased corticosterone plasma concentrations (both P < 0.05). Conclusion Deficiency or functional blockade of TLR2 is associated with increased survival and improved neurofunctional outcome in a mouse model of CA/CPR. Thus, TLR2 inhibition could provide a novel therapeutic approach for reducing mortality and morbidity after cardiac arrest and cardiopulmonary resuscitation.