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Clostridioides difficile infections have become a major challenge in medical facilities. The bacterium is capable of spore formation allowing the survival of antibiotic treatment. Therefore, research on the physiology of C. difficile is important for the development of alternative treatment strategies. In this study, we investigated eight putative flavodoxins of C. difficile 630. Flavodoxins are small electron transfer proteins of specifically low potential. The unusually high number of flavodoxins in C. difficile suggests that they are expressed under different conditions. We determined high transcription levels for several flavodoxins during the exponential growth phase, especially for floX . Since flavodoxins are capable of replacing ferredoxins under iron deficiency conditions in other bacteria, we also examined their expression in C. difficile under low iron and no iron levels. In particular, the amount of fldX increased with decreasing iron concentration and thus could possibly replace ferredoxins. Moreover, we demonstrated that fldX is increasingly expressed under different oxidative stress conditions and thus may play an important role in the oxidative stress response. While increased fldX expression was detectable at both RNA and protein level, CD2825 showed increased expression only at mRNA level under H 2 O 2 stress with sufficient iron availability and may indicate hydroxyl radical-dependent transcription. Although the exact function of the individual flavodoxins in C. difficile needs to be further investigated, the present study shows that flavodoxins could play an important role in several physiological processes and under infection-relevant conditions., Importance: The gram-positive, anaerobic, and spore-forming bacterium Clostridioides difficile has become a vast problem in human health care facilities. The antibiotic-associated infection with this intestinal pathogen causes serious and recurrent inflammation of the intestinal epithelium, in many cases with a severe course. To come up with novel targeted therapies against C. difficile infections, a more detailed knowledge on the pathogen's physiology is mandatory. Eight putative flavodoxins, an extraordinarily high copy number of this type of small electron transfer proteins, are annotated for C. difficile . Flavodoxins are known to be essential electron carriers in other bacteria, for instance, during infection-relevant conditions such as iron limitation and oxidative stress. This work is a first and comprehensive overview on characteristics and expression profiles of the putative flavodoxins in the pathogen C. difficile ., Competing Interests: The authors declare no conflict of interest.

The anaerobic bacterium Clostridioides difficile represents one of the most problematic pathogens, especially in hospitals. Dysbiosis has been proven to largely reduce colonization resistance against this intestinal pathogen. The beneficial effect of the microbiota is closely associated with the metabolic activity of intestinal microbes such as the ability to transform primary bile acids into secondary ones. However, the basis and the molecular action of bile acids (BAs) on the pathogen are not well understood. We stressed the pathogen with the four most abundant human bile acids: cholic acid (CA), chenodeoxycholic acid (CDCA), deoxycholic acid (DCA) and lithocholic acid (LCA). Thin layer chromatography (TLC), confocal laser scanning microscopy (CLSM), and electron microscopy (EM) were employed to track the enrichment and destination of bile acids in the bacterial cell. TLC not only revealed a strong accumulation of LCA in C. difficile , but also indicated changes in the composition of membrane lipids in BA-treated cells. Furthermore, morphological changes induced by BAs were determined, most pronounced in the virtually complete loss of flagella in LCA-stressed cells and a flagella reduction after DCA and CDCA challenge. Quantification of both, protein and RNA of the main flagella component FliC proved the decrease in flagella to originate from a change in gene expression on transcriptional level. Notably, the loss of flagella provoked by LCA did not reduce adhesion ability of C. difficile to Caco-2 cells. Most remarkably, extracellular toxin A levels in the presence of BAs showed a similar pattern as flagella expression. That is, CA did not affect toxin expression, whereas lower secretion of toxin A was determined in cells stressed with LCA, DCA or CDCA. In summary, the various BAs were shown to differentially modify virulence determinants, such as flagella expression, host cell adhesion and toxin synthesis. Our results indicate differences of BAs in cellular localization and impact on membrane composition, which could be a reason of their diverse effects. This study is a starting point in the elucidation of the molecular mechanisms underlying the differences in BA action, which in turn can be vital regarding the outcome of a C. difficile infection., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Metzendorf, Lange, Lainer, Schlüter, Dittmann, Paul, Troitzsch and Sievers.)

The human pathogen Clostridioides difficile has evolved into the leading cause of nosocomial diarrhea. The bacterium is capable of spore formation, which even allows survival of antibiotic treatment. Although C. difficile features an anaerobic lifestyle, we determined a remarkably high oxygen tolerance of the laboratory reference strain 630Δ erm A mutation of a single nucleotide (single nucleotide polymorphism [SNP]) in the DNA sequence (A to G) of the gene encoding the regulatory protein PerR results in an amino acid substitution (Thr to Ala) in one of the helices of the helix-turn-helix DNA binding domain of this transcriptional repressor in C. difficile 630Δ erm PerR is a sensor protein for hydrogen peroxide and controls the expression of genes involved in the oxidative stress response. We show that PerR of C. difficile 630Δ erm has lost its ability to bind the promoter region of PerR-controlled genes. This results in a constitutive derepression of genes encoding oxidative stress proteins such as a rubrerythrin ( rbr1 ) whose mRNA abundance under anaerobic conditions was increased by a factor of about 7 compared to its parental strain C. difficile 630. Rubrerythrin repression in strain 630Δ erm could be restored by the introduction of PerR from strain 630. The permanent oxidative stress response of C. difficile 630Δ erm observed here should be considered in physiological and pathophysiological investigations based on this widely used model strain. IMPORTANCE The intestinal pathogen Clostridioides difficile is one of the major challenges in medical facilities nowadays. In order to better combat the bacterium, detailed knowledge of its physiology is mandatory. C. difficile strain 630Δ erm was generated in a laboratory from the patient-isolated strain C. difficile 630 and represents a reference strain for many researchers in the field, serving as the basis for the construction of insertional gene knockout mutants. In our work, we demonstrate that this strain is characterized by an uncontrolled oxidative stress response as a result of a single-base-pair substitution in the sequence of a transcriptional regulator. C. difficile researchers working with model strain 630Δ erm should be aware of this permanent stress response., (Copyright © 2021 Troitzsch et al.)

Background: Recent publications have raised concerns of reduced susceptibilities of clinical bacterial isolates towards biocides. This study presents a comparative investigation of the susceptibility of livestock-associated Methicillin-resistant Staphylococcus aureus (LA-MRSA), hospital-acquired MRSA (HA-MRSA) and community-aquired MRSA (CA-MRSA) to the commonly used antiseptics chlorhexidine (CHX), octenidine (OCT), polyhexanide (PHMB), PVP-iodine (PVP-I) and triclosan (TCX) based on internationally accepted standards., Methods: In total, 28 (18 LA-, 5 HA- and 5 CA) genetically characterized MRSA strains representing a broad spectrum of hosts, clonal complexes and spa-types, as well as the reference methicillin-sensitive Staphylococcus aureus (MSSA) strain ATCC 6538, were selected. Minimal inhibitory concentration (MIC) and minimal microbicidal concentration (MBC) were determined in accordance with DIN 58940-7, 58940-8 and DIN EN ISO 20776-1. The microbicidal efficacy was determined in accordance with DIN EN 1040., Results: Results from the MIC/MBC and quantitative suspension tests revealed differences between antiseptic substances but not between epidemiological groups of MRSA strains. OCT and PHMB were the most active substances with a minimal MIC of 1 mg/L, followed by CHX (2 mg/L), TCX (32 mg/L) and finally PVP-I (1024 mg/L). The MSSA reference strain showed a tendency to a higher susceptibility compared to the MRSA strains., Conclusions: This investigation of the susceptibility of a range of LA-, HA- and CA-MRSA strains using standardized conditions gave no indication that LA-MRSA strains are less susceptible to commonly used antiseptics compared to HA- and CA-MRSA strains., Competing Interests: Competing interestsPeter Pfaff is an employee of BBraun AG GmbH (Melsungen, Germany). The antiseptic compounds CHX and PHMB are part of some of the products of BBraun AG GmbH. The other authors declare that they have no competing interests. None of the authors holds stock or options in BBraun AG GmbH.

Clostridioides difficile is an intestinal human pathogen that uses the opportunity of a depleted microbiota to cause an infection. It is known, that the composition of the intestinal bile acid cocktail has a great impact on the susceptibility toward a C. difficile infection. However, the specific response of growing C. difficile cells to diverse bile acids on the molecular level has not been described yet. In this study, we recorded proteome signatures of shock and long-term (LT) stress with the four main bile acids cholic acid (CA), chenodeoxycholic acid (CDCA), deoxycholic acid (DCA), and lithocholic acid (LCA). A general overlapping response to all tested bile acids could be determined particularly in shock experiments which appears plausible in the light of their common steroid structure. However, during LT stress several proteins showed an altered abundance in the presence of only a single or a few of the bile acids indicating the existence of specific adaptation mechanisms. Our results point at a differential induction of the groEL and dnaKJgrpE chaperone systems, both belonging to the class I heat shock genes. Additionally, central metabolic pathways involving butyrate fermentation and the reductive Stickland fermentation of leucine were effected, although CA caused a proteome signature different from the other three bile acids. Furthermore, quantitative proteomics revealed a loss of flagellar proteins in LT stress with LCA. The absence of flagella could be substantiated by electron microscopy which also indicated less flagellated cells in the presence of DCA and CDCA and no influence on flagella formation by CA. Our data break down the bile acid stress response of C. difficile into a general and a specific adaptation. The latter cannot simply be divided into a response to primary and secondary bile acids, but rather reflects a complex and variable adaptation process enabling C. difficile to survive and to cause an infection in the intestinal tract.

Clostridioides difficile is the major pathogen causing diarrhea following antibiotic treatment. It is considered to be a strictly anaerobic bacterium, however, previous studies have shown a certain and strain-dependent oxygen tolerance. In this study, the model strain C. difficile 630Δerm was shifted to micro-aerobiosis and was found to stay growing to the same extent as anaerobically growing cells with only few changes in the metabolite pattern. However, an extensive change in gene expression was determined by RNA-Seq. The most striking adaptation strategies involve a change in the reductive fermentation pathways of the amino acids proline, glycine and leucine. But also a far-reaching restructuring in the carbohydrate metabolism was detected with changes in the phosphotransferase system (PTS) facilitated uptake of sugars and a repression of enzymes of glycolysis and butyrate fermentation. Furthermore, a temporary induction in the synthesis of cofactor riboflavin was detected possibly due to an increased demand for flavin mononucleotid (FMN) and flavin adenine dinucleotide (FAD) in redox reactions. However, biosynthesis of the cofactors thiamin pyrophosphate and cobalamin were repressed deducing oxidation-prone enzymes and intermediates in these pathways. Micro-aerobically shocked cells were characterized by an increased demand for cysteine and a thiol redox proteomics approach revealed a dramatic increase in the oxidative state of cysteine in more than 800 peptides after 15 min of micro-aerobic shock. This provides not only a catalogue of oxidation-prone cysteine residues in the C. difficile proteome but also puts the amino acid cysteine into a key position in the oxidative stress response. Our study suggests that tolerance of C. difficile towards O 2 is based on a complex and far-reaching adjustment of global gene expression which leads to only a slight change in phenotype., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Objective: The postoperative systemic inflammatory response after cardiopulmonary bypass (CPB) is still an undesirable side-effect after cardiac surgery. It is most likely caused by blood contact with foreign surfaces and by the surgical trauma itself. However, the recirculation of activated shed mediastinal blood is another main cause of blood cell activation and cytokine release. Minimal invasive extracorporeal circulation (MiECC) comprises a completely closed circuit, coated surfaces and the separation of suction blood. We hypothesized that MiECC, with separated cell saved blood, would induce less of a systemic inflammatory response than MiECC with no cell-saver. The aim of this study was, therefore, to investigate the impact of cell washing shed blood from the operating field versus direct return to the ECC on the biomarkers for systemic inflammation., Material and Methods: In the study, patients with MiECC and cell-saver were compared with the control group, patients with MiECC and direct re-transfusion of the drawn blood shed from the surgical field., Results: High amounts of TNF-α (+ 120% compared to serum blood) were found in the shed blood itself, but a significant reduction was demonstrated with the use of a cell-saver (TNF-α ng/l post-ECC 10 min: 9.5±3.5 vs. 19.7±14.5, p<0.0001). The values for procalcitonin were not significantly increased in the control group (6h: 1.07±3.4 vs. 2.15±9.55, p=0.19) and lower for C-reactive protein (CRP) (24h: 147.1±64.0 vs.134.4±52.4 p=0.28)., Conclusion: The use of a cell-saver and the processing of shed blood as an integral part of MiECC significantly reduces the systemic cytokine load. We, therefore, recommend the integration of cell-saving devices in MiECC to reduce the perioperative inflammatory response.

Purpose: This study addresses the effect of short myocardial ischemia on inhibitory effect of ATP for mitochondrial cytochrome c oxidase (CytOx) activity in myocardium and subsequent hemodynamic alterations. The activity of CytOx is inhibited by ATP (primary substrate control). This additional mechanism was proposed to be switched off at higher mitochondrial membrane potential values in case of stress. The ATP-dependent allosteric enzyme inhibition (second respiratory control) is suggested to reduce the formation of reactive oxygen species and thus is pivotal for cytoprotection. This report addresses the possible involvement of this mechanism in case of myocardial preconditioning., Methods: Rat hearts were perfused in a Langendorff system (n = 5 each group). The first two groups underwent short recurrent ischemic periods (three times 5 min) and subsequent high or low reperfusion for 40 min. Besides four control groups, hearts were exposed to an ischemia of 15 min and high flow reperfused for 30 min, in addition. Hemodynamic data were evaluated in parallel. Mitochondria were separated for the polarographic respiration measurements in the presence of ADP or ATP, respectively. Phosphorylation patterns of the CytOx subunits were studied by immunoblotting with P-Ser, P-Thr, and P-Tyr antibodies., Results: Short recurrent episodes of ischemia result in an ATP-dependent inhibition of CytOx. Electrophoretic analysis and blotting techniques reveal different phosphorylation patterns of the enzyme. Frequent short-lasting ischemic impacts and subsequent increased coronary flow seem to be essential for this effect., Conclusion: The procedure of preconditioning is likely to be dependent on the mechanism of ATP-dependent inhibition of CytOx activity.

We hypothesized that pretreatment with single-dose cyclosporine (CsA) prevents alterations and improves tissue oxygen and mitochondrial cytochrome oxidase redox (CytOx) state in skeletal muscle ischemia and reperfusion-reoxygenation (I/R). Latissimus dorsi muscle was prepared and mobilized in New Zealand white rabbits. Ischemia was induced for 4 h, followed by 2 h of reperfusion. The animals were randomized to receive a 60-mg/kg intravenous bolus of CsA (CsA group, n = 10) or physiologic saline (control, n = 10) at 10 min before ischemia onset. Muscle tissue oxygen tension (PtO(2)) and mitochondrial CytOx were measured during I/R simultaneously. High-energy phosphate (HEP) levels were determined using high-field (31)P magnetic resonance spectroscopy. Mitochondrial viability index and wet-to-dry ratio were used to assess the tissue viability between groups. Decreases in tissue oxygen levels and CytOx were slower during ischemia in the CsA group in comparison to control group, also the loss of phosphocreatine and adenosine triphosphate depletion. After ischemia, recovery of tissue oxygen, mitochondrial CytOx, and HEP was delayed in controls. Tissue PtO2 in the CsA group (P < 0.05) was significantly higher compared with that in the control group after I/R. Mitochondrial CytOx was also improved in the CsA group (P < 0.01 vs. control). Muscle HEP levels (phosphocreatine, adenosine triphosphate) were significantly preserved in the CsA group versus the control group (P < 0.01, P < 0.05). Mitochondrial viability index and wet-to-dry ratio confirmed significantly preserved tissue and lower edema formation in the CsA group. The pretreatment with single-dose CsA prevents alterations and improves tissue oxygenation and mitochondrial oxidation in skeletal muscle I/R.

Background: In this study, the microvascular tissue oxygenation and oxidative muscle metabolism during graded hypoxia and reoxygenation were examined in a rabbit model by near-infrared (NIR) spectroscopy and correlated with high-energy phosphates measured by (31)P nuclear magnetic resonance (NMR) spectroscopy., Materials and Methods: Graded hypoxia was performed in a New Zealand rabbit model (n = 20, 2.0 ± 0.4 kg) by a stepwise reduction of the fraction of inspired oxygen (FiO(2)) from 0.3 to 0.05 (intervention group versus control group). Recovery and reoxygenation were achieved using FiO(2) of 0.3. A noninvasive NIR spectroscopy sensor and NMR probe was positioned on the surface of the prepared pedicled latissimus dorsi muscle. Microvascular tissue oxygenation (oxyhemoglobin, HbO(2); deoxyhemoglobin, HHb) and redox state of cytochrome oxidase (CytOx) were measured by NIR spectroscopy and correlated with standard values of oxidative muscle metabolism (phosphocreatine, PCr; adenosine triphosphate, ATP) measured by time-resolved (31)P NMR spectroscopy (4.7T)., Results: Significant correlation was found between PCr and HbO(2) (r = 0.85, P < 0.001) and HHb (r = -0.75, P < 0.001). β-ATP levels correlated significantly with CytOx (r = 0.87, P < 0.001)., Conclusions: The data suggest that changes in high-energy phosphates (PCr- and ATP-levels) correlate closely with microvascular tissue oxygenation (HbO(2), HHb, CytOx) measured by NIR spectroscopy., (Copyright © 2012 Elsevier Inc. All rights reserved.)

Background: There is currently no efficient and reliable clinical means of assessing the degree of ischemia- and reperfusion-associated damage in microvascular transplants. The objective was to study correlation of tissue oxygen tension measurements with tissue oxygen saturation, cytochrome oxidase redox state, and tissue viability., Materials and Methods: Latissimus dorsi muscle was dissected and mobilized in New Zealand white rabbits (n = 30, 2.5 ± 0.5 kg). Muscles were exposed to warm ischemia in two groups with either 4 or 6 h, followed by reperfusion. Tissue PO(2) was measured with a miniature intramuscular oxygen sensor (Licox microprobe; Integra Neurosciences, Ratingen, Germany) all along with tissue hemoglobin saturation (rSO(2)) and cytochrome oxidase aa3 redox state (CytOx) by in vivo near-infrared spectroscopy. Linear correlation was performed between tissue PO(2) and rSO(2), CytOx and tissue viability., Results: After ischemia and reperfusion, tissue PO(2) and CytOx recovery was significantly decreased in both groups compared with control (4 h: P < 0.05; 6 h: P < 0.01). Significant correlations between changes in tissue PO(2) and rSO(2) (r = 0.92; P < 0.01), CytOx (r = 0.90; P < 0.01), wet-to-dry ratio (r = -0.97; P < 0.01), and mitochondrial viability index (r = 0.97; P < 0.01) were found., Conclusions: Tissue oxygen tension measured with microprobes correlated closely with tissue oxygenation, cellular oxygen utilization, and the extent of ischemia reperfusion injury., (Copyright © 2011 Elsevier Inc. All rights reserved.)

Simultaneous measurements of extracellular pH, potassium (K(+)), and calcium (Ca(2+)) activity might be indicative of myocardium vitality or ischemia. Ten consecutive patients undergoing elective coronary artery bypass grafting were studied. Epicardial extracellular pH, potassium, and calcium were measured by a miniaturized disposable multi-sensor probe. Blood gases and electrolytes were derived with measurements of arterial and mixed venous blood samples at intervals during surgery. The mean epicardial baseline levels for pH in all patients were 8.04+/-0.22 arbitrary units (AU) for the right ventricle (RV) and 8.03+/-0.21 AU for the left ventricle (LV); for Ca(2+) 0.23+/-0.07 mmol/l (RV) and 0.20+/-0.10 mmol/l (LV); and for K(+) 4.54+/-1.51 mmol/l (RV) and 4.38+/-0.57 mmol/l (LV). Before ischemia, epicardial pH was moderately (p<0.05), and K(+), and Ca(2+) were closely correlated (p<0.001) with blood values. During reperfusion, epicardial measurements were weakly correlated (p<0.001) with blood values for pH, venous K(+) and Ca(2+), but moderately correlated with arterial K(+) and Ca(2+) (p<0.01). The measurements indicated intraoperative episodes of ischemia and reperfusion with reproducible trends of extracellular pH, K(+), and Ca(2+), which results in electrolyte patterns applicable for detecting inadequate myocardial protection during cardiac surgery in patients.

Effective myocardial oxygen supply should not be compromised during cardiac surgery as it is essential to avoid circulatory and cardiac dysfunction. Local measurement of myocardial oxygen partial pressure (pO2) was therefore introduced into the operative monitoring of myocardial ischemia. The aim of the present study was to assess whether myocardial oxygen partial pressure correlates with the content of high energy phosphates (HEPs). Seven male rabbits were examined in parallel with measurement of myocardial pO2 by an implanted Clark electrode and 31phosphorus-NMR spectroscopy. The ventilatory management established hyperoxygenation followed by systemic hypoxia with hypercapnia for 20 min. Additionally, analysis of end-expiratory gas composition in combination with blood gas analysis was performed simultaneously, and hemodynamic parameter was recorded. Under hypoxic conditions the cardiovascular system was severely compromised, whereas the myocardial pO2 was only moderately impaired (pO2M 45.0+/-16.0 mm Hg). Immediately before cardiac arrest, low values of arterial and venous pO2 were found (17.6+/-6.0 and 12.9+/-6.1 mm Hg). In contrast to near normal myocardial pO2, HEP content in the myocardium was considerably reduced and inorganic phosphorus was increased. Artificial ventilation leading to systemic hypoxia and eventually circulatory arrest resulted in almost normal myocardial pO2 but severely compromised HEP content. This somewhat unexpected finding requires further clarification, but is in accordance with findings reported previously where regulatory mechanisms have been shown to play a role in the pathophysiology of severe hypoxic conditions such as those for cellular oxygen delivery and demand, P/O coupling and finally control of HEP production facilitating the interaction between respiratory chain and myoglobin oxygen transport.

Background: The capacity of heat stress induction to improve myocardial tolerance against ischemia is well known. We investigated cardiac energy metabolism after hsp 72(+)/73(+) induction in isolated perfused neonatal rabbit hearts subjected to prolonged cold cardioplegic ischemia., Methods: Hearts from neonatal rabbits were excised, isolated perfused and arrested by 2-h cold cardioplegic ischemia. Rectal temperature of eight neonatal rabbits was raised to 42.0 to 42.5 degrees C for heat shock protein expression in a whole body water bath for 15 min before the onset of arrest. Another set of eight rabbits without hyperthermia pretreatment served as control. Recovery of left ventricle function was assessed by aortic flow, cardiac output, and max dP/dt. Status of high-energy phosphates was measured by (31)phosphorus nuclear magnetic resonance-spectroscopy., Results: Immunoblot analysis revealed clear hsp 72+/73+ induction after a brief period of systemic hyperthermia. Heat stress pretreatment resulted in a better recovery of left ventricular function (aortic flow and cardiac output improvement P < 0.05, max dP/dt P < 0.01) than in controls at 60 min after reperfusion. During ischemia and reperfusion, myocardial energy metabolism was better preserved in hearts after hsp induction as a consequence of increased gamma-, alpha-, and beta-ATP as well as phosphocreatine-values over controls. The ischemia-induced pH-decrease was attenuated., Conclusion: These data contribute to the evidence of heat stress mediated beneficial effects on functional myocardial recovery and improved cardiac energy metabolism after prolonged cold cardioplegic ischemia. More importantly, the attenuation of ischemic pH reduction and better restoration suggest an involvement of mitochondrial membrane potential alterations.

Conflicting results have been reported with regard to the neuroprotective effects of steroid treatment with cardiopulmonary bypass (CPB) and deep hypothermic circulatory arrest (DHCA). We evaluated the mode and severity of neuronal cell injury in neonatal piglets after prolonged DHCA and the possible neuroprotective effect of systemic pretreatment (>6 h before surgery) with large-dose methylprednisolone (MP). Nineteen neonatal piglets (age, <10 days; weight, 2.1 +/- 0.5 kg) were randomly assigned to 2 groups: 7 animals were pretreated with large-dose systemic MP (30 mg/kg) 24 h before surgery, and 12 animals without pharmacological pretreatment (saline) served as control groups. All animals were connected to full-flow CPB with cooling to 15 degrees C and 120 min of DHCA. After rewarming to 38.5 degrees C with CPB, animals were weaned from CPB and survived 6 h before they were killed, and the brain was prepared for light and electron microscopy, immunohistochemistry, and TUNEL-staining. Quantitative histological studies were performed in hippocampus, cortex, cerebellum, and caudate nucleus. Systemic pretreatment with large-dose MP lead to persistent hyperglycemia but no significant changes of cerebral perfusion. Necrotic and apoptotic neuronal cell death were detected in all analyzed brain regions after 120 min of DHCA. In comparison to the control group, large-dose pretreatment with systemic MP lead to an increase of necrotic neuronal cell death and induced significant neuronal apoptosis in the dentate gyrus of the hippocampus (P = 0.001). In conclusion, systemic pretreatment with large-dose MP fails to attenuate neuronal cell injury after prolonged DHCA and induces regional neuronal apoptosis in the dentate gyrus.

Background: Recent studies have shown a relationship between alterations in tissue oxygen metabolism and cellular changes following ischemia and reperfusion, such as energy store depletion and intracellular acidosis. The aim of this study was to evaluate the relationship between tissue energy metabolism and intramuscular tissue oxygen tension in the mobilized latissimus dorsi muscle., Material and Methods: The latissimus dorsi muscle was raised in New Zealand white rabbits (n = 10, 2.5 +/- 0.5 kg). During 4 h of ischemia and 2 h of reperfusion, the intramuscular tissue oxygen tension (Licox PO2-microcatheter probe) and the status of phosphorylated muscle energy metabolites were measured using a high-field 31P-NMR spectrometer. Linear correlation was performed between 31P-NMR data and tissue oxygen tension readings., Results: The tissue oxygen tension (PO2) values correlated significantly with phosphocreatine (PCr) (r = 0.96, P < 0.001), beta-adenosin triphosphate (beta-ATP) (r = 0.64, P <0.01), and intracellular pH (r = 0.82, P <0.001)., Conclusions: On the basis of these findings, we conclude that the data provided by tissue oxygen tension measurement offer a real time minimally invasive estimate of muscle oxidative metabolism during ischemia and reperfusion.

Myocardial hypoxia is accompanied by increased epicardial potassium activity. In this experimental study in rabbits (n = 13), with ion-selective probes, we correlated our findings with an exact evaluation of myocardial oxygen tension, hemodynamic data and arterial and venous blood gas analysis. The epicardial potassium activity had good correlation with these parameters (pm = -0.95, pco = -0.93 and pa = -0.72, pv = -0.96, respectively). Therefore, the clinical use of these electrodes can be recommended for continuous monitoring in intensive care units.

Percutaneous sclerotherapy of orbital low-flow vascular malformations requires precise procedural guidance. For the treatment of a patient with an orbital venolymphatic malformation, we sought to optimize guidance by combining navigation assistance for needle placement with intralesional contrast medium injection for assessment of venous drainage. By using a surgical navigation system (Vector Vision, BrainLAB, Munich, Germany), multiplanar target lesion visualization was performed after fusion of CT and MR imaging data, which allowed precise puncture planning.

Background: Recent findings indicate that molecular chaperones actively participate in myocardial cytoprotection. Moreover, ischemic tolerance can be induced in humans by brief ischemic events. Therefore, we investigated patients with severe angina attacks before coronary artery bypass grafting. We focused on appearance of anti-hsp70 antibodies as an immunologic response to heat shock protein induction by ischemia followed up by hemodynamic measurements perioperatively. We correlated these clinical findings with the presence of antibodies against hsp70 and the antioxidative capacity of patients' sera., Methods: Thirty-five consecutive patients with coronary artery disease scheduled for coronary artery bypass grafting were included. Seventeen patients had severe angina, and 18 patients suffered from chronic stable angina preoperatively. In the patients' sera, antibodies against hsp70 were detected by enzyme-linked immunosorbent assay, and antioxidative capacity was detected using the chromogen assay. Cardiac output and pulmonary capillary wedge pressure were measured using a thermodilution catheter. We also evaluated C-reactive protein and creatine kinase MB isoenzyme, and performed a conventional leukocyte count., Results: The sera of the 17 patients with severe angina attacks before surgery contained antibodies against hsp70 and a low antioxidative capacity. The interval between a severe angina attack and anti-hsp70 antibody titer are inversely correlated. These patients had better cardiac output and lower pulmonary capillary wedge pressure values after surgery., Conclusions: Severe angina before cardiac surgery coincided with an improved outcome as measured by hemodynamic variables as compared with chronic stable angina. This finding correlated significantly with a low antioxidative capacity and the presence of antibodies against hsp70. These pathophysiologic mechanisms might therefore play a role in myocardial protection.

Ossifying fibroma is a benign fibro-osseous tumor commonly affecting the craniofacial bones. It is considered to be a locally aggressive and quickly expansible bone lesion. Because of its aggressive nature and high recurrence rate, early detection and complete surgical removal are essential. Usually, these lesions are excised extensively by craniectomy, and bone loss is reconstructed by cranioplasty using acrylic resin or titanium implants. Alternatively, in the management of skull-ossifying fibroma, an image-guided technique using surgical navigation may provide precise information about localization, enabling complete removal, thereby operating with minimal exposure and within narrow resection borders and avoiding significant bone deformity. A 39-year-old male patient with a history of renal cell carcinoma was admitted to our hospital because a radionuclide scintigraphic bone scan revealed increased uptake in a small area located at the left lateral skull bone. The high-resolution computed tomography scan showed that the lesion was located inside the diploe, destroying the inner table of the calvarium. The patient underwent minimally invasive bone lesion removal using an interactive image-guided approach. Complete resection of the neoplastic lesion was achieved. The histopathological examination revealed an ossifying fibroma. The postoperative course was uneventful, and the patient was discharged 3 days after intervention. To date, there has been no evidence of local recurrence. Interactive multimodal planning and intraoperative image guidance offer an interesting approach for biopsy and minimally invasive removal of small ossifying fibroma lesions of the skull, especially in less accessible locations.

Objectives/hypothesis: Image-data-based surgical navigation is used as a helpful device in the operating room to localize critical structures with a high degree of accuracy. It also enables physicians to plan therapeutic performance. Because it relies on preoperatively acquired computed tomography (CT) or magnetic resonance imaging (MRI) data, there is restricted access for navigation of surgical instruments in areas that show motion uncorrelated with radiologic data. Thus, in the case of moveable structures, for example the lower jaw, navigational procedures could not yet be applied., Study Design: We introduce a new technique using individualized intermaxillary splints that fix the mandible in a reproducible aboccluded position at the time of image-data acquisition and surgery., Methods: Different manufacturing processes were investigated. The feasibility of uni- and bilateral intermaxillary splints was studied under clinical conditions in four patients during different procedures in the mandibular and oropharyngeal regions., Results: The manufacturing of the splints showed was easily performed in a short time. With bilateral fixation, there was a high anatomic target precision of 1.6 to 2.3 mm., Conclusions: The use of bilateral intermaxillary splints that fix the patient's mandible in a reproducible aboccluded position permits an image-data-based navigated surgical approach to the oropharyngeal and mandibular regions.

Laser-induced interstitial thermal therapy (LITT) is a proven minimally invasive surgical technique for the treatment of haemangiomas and vascular malformations, and various tumours. The intra-lesion application of thermal energy destroys regional tissue. The percutaneous placement of the laser fibre for photocoagulation is done without the benefit of direct visual control. Image-data-based LITT was performed in patients (five procedures) with extensive venous malformations in the maxillofacial area. The system comprised a specially developed Nd:YAG laser fibre introducer set used in conjunction with fused high-resolution computed tomography, and magnetic resonance image-data--based surgical navigation. In all cases, follow-up examination clearly showed a diminishment in tumour volume, and all patients reported significant subjective improvement. The results suggest that navigation-guided LITT can be performed safely, preserving vital structures from collateral thermal damage.

Introduction: Recent innovations in laser scanning technology provide a potentially useful tool for three-dimensional surface registration for image-guided surgery. The purpose of this study is to evaluate the clinical reliability of this technique in oral and maxillofacial surgical procedures using image-guided navigation., Methods: In an experimental step, a stable anthropomorphic skull model with prelabeled markers was scanned and registered with laser surface scanning (z-touch, BrainLAB) and marker- based algorithms. The registration protocol was then repeated 25-times. Root mean square error (RMSE) and target difference values were compared for their suitability for this application. Twelve patients with different indications for oral and maxillofacial surgery were planned for image-guided surgery using a passive infrared surgical navigation system (VectorVision, BrainLAB). Preoperative computed tomography (CT) scans were carried out with newest 16-line multisclice CT-scanner (Siemens Somatom Sensation 16). The new markerless laser surface scanning technique was applied in all intraoperative patient registrations. Registration error was noted. The clinical application accuracy was determined for anatomical landmark localization deviation., Results: In the experimental protocol a mean registration error (RMSE) or target difference of 1.3 (0.14) or 2.08 (0.49) mm for laser scanning and 0.38 (0.01) or 0.99 (0.15) mm for marker registration was found. The differences for RMSE and target localization were statistically significant (p < 0.005). Furthermore, a strong correlation between RMSE and target difference was found for laser scanning (r = 0.96) and marker registration (r = 0.95). During various clinical procedures involving oral and maxillofacial surgery, the overall error of the registration procedure determined as RMSE was 1.21 (0.34) mm. Intraoperatively, the mean clinical application accuracy was found to be 1.8 (0.5) mm., Conclusion: Three-dimensional laser surface scanning technique may be a interesting and useful approach to register the patient for image-guided procedures, particularly during oral and craniomaxillofacial surgery.

Objective: Brain injury and altered psychomotor development in infants, children and adults after cardiac surgery using cardiopulmonary bypass (CPB) and deep hypothermic circulatory arrest (DHCA) is still a matter of concern. Early diagnosis and identification of brain injury that has occurred or is ongoing by measurement of biochemical markers in serum may have diagnostic and prognostic value. The aim of the experimental studies in an animal model was therefore to investigate the release patterns of astroglial and neuronal markers in serum and to determine the morphological and immunohistochemical changes in the brain of animals undergoing similar perfusion conditions of CPB and a period of DHCA., Methods: Fourteen New Zealand rabbits, (weight, 3.1 +/- 0.25 kg) were anesthetized, intubated and mechanically ventilated. Four animals were sham operated and served as controls. After median sternotomy the animals were connected to CPB by cannulation of the aorta and right atrium. Full flow CPB (200-250 ml/kg/min) was initiated to achieve homogeneous systemic cooling. Circulatory arrest of 60 minutes was induced when rectal and nasopharyngeal temperature of 14 degrees C was achieved. After rewarmed reperfusion and establishment of stable cardiac ejection the animals were weaned from CPB and monitored for 6 hours. Then the animals were killed, the brain was immediately removed and cut in standardized sections. These were fixated, embedded in paraffin and stained for further quantitative histological studies. In the brain astrocyte reactivity for S-100B was assessed immunocytochemically (DPC Immustain Los Angeles, USA). Monoclonal mouse anti-human neurospecific enolase (NSE) antibody was used for the localization of NSE in the fixed and paraffin embedded brain (NSE-DAKO, H14). The concentrations of S-100B protein and neurospecific enolase (NSE) in the serum were analyzed using a commercially available immunoluminometric assay (LIA-mat, Sangtec 100, Byk-Sangtec). Immunospecific monoclonal anti-parvalbumin antibody was used for the detection of parvalbumin in the brain. Serum concentrations of parvalbumin were analyzed using a newly developed ELISA method., Results: In all experimental animals a significant increase of the serum concentration of the astroglial protein S-100B was found immediately after reperfusion and the termination of CPB. In contrast the serum levels of the neuronal proteins parvalbumin and NSE were not increased, but rather decreased. Light microscopy and electron microscopy revealed perivascular astrocytic swelling and minor neuronal cell injury. In comparison to the sham operated animals, increased immunohistochemical staining of S-100B was found. This increased reactivity of S100B antibody was found in the astrocytic processes with immediate connection to the perivascular space and around the perivascular oedema. The immunocytochemical stainings for NSE and parvalbumin in the neuronal cells was not different from that of sham-operated animals and indicated well preserved neurons.

Early detection of myocardial ischaemia is a central problem in cardiological and cardiosurgical intensive care. A new approach is the use of ion-selective electrodes implanted directly on the myocardium, enabling detection of increased potassium activity as an indication of general hypoxia. After a comprehensive study of the electrode parameters, an animal experiment was carried out, in which it was found that respiration-induced hypoxia resulted in an increase in epicardial potassium activity (p < 0.01). Blood gas analysis performed simultaneously revealed reduced arterial pO2, but no acidosis. Haemodynamic data evidenced hypoxic depression of circulatory parameters. Histological examinations of the myocardium beneath the electrodes revealed typical lymphocytic infiltration. Electron microscopy demonstrated crystolysis in the mitochondria as an early sign of hypoxia, thus confirming the sensitivity of these electrodes. This underscores the potential of ion-selective electrodes for the detection of myocardial ischaemia, and they should now be investigated in the clinical setting.

Advances in intra-operative imaging and the development of new minimally invasive techniques are having an ever greater impact on modern surgery. Mobile CT scanners in the operating room is a new technique that permits image-guided surgery, and helps minimize postoperative complications. We report on our initial experience with intraoperative CT scanning during surgery on patients suffering lateral midface trauma. A mobile CT unit, the Tomoscan M (Philips, Utrecht, Netherlands) set up in the operating room, was evaluated in 6 patients with zygomatic bone fractures. The patients were placed on the CT scanner table, which is detachable from the gantry. The unit is powered by batteries charged from an ordinary ring mains supply via a conventional plug. The CT images obtained were of good quality in all cases. No technical problems were observed during surgery. Using repeat CT scans, the procedure also permits accurate intraoperative monitoring of the anatomical repositioning of the bone fragments, and accurate implantation. No intraoperative or early postoperative complications were observed. This new technical aid ensures highly accurate reduction of the bone fragments, and minimizes the need for reoperation. High-quality intraoperative imaging with surgical navigation increase surgical outcome, and expand the spectrum of minimally invasive surgery.

Background: This study was undertaken to investigate the physiological effects of cardiopulmonary bypass (CPB) and deep hypothermic circulatory arrest (DHCA) on cerebral oxygen metabolism estimated by near-infrared spectroscopy (NIRS)., Methods: Ten newborn piglets (2.1 to 2.6 kg) were monitored with right frontal NIRS; the right jugular bulb was cannulated for intermittent sampling of jugular venous blood. All animals underwent CPB, cooling to a core temperature below 15 degrees C, 60 minutes of DHCA followed by subsequent reperfusion and rewarming. Continuously recorded NIRS data and intermittent jugular venous blood values were compared., Results: NIRS performance was examined over the jugular venous oxygen saturation (SjvO2) range of 40 to 98 %, a linear correlation was found between SjvO2 and NIRS-derived regional cerebral oxygen saturation (rSO2) (r = 0.91, p < 0.001). A correlation was observed between the cellular oxidation NIRS-parameter cytochrome oxidase aa3 (CytOx) slope during the DHCA period in relation to rectal and nasopharyngeal temperature immediately before the onset of DHCA (r = 0.75 and 0.85, p < 0.001)., Conclusions: This study suggests that NIRS-measured hemoglobin oxygenation parameters may reflect functional changes in cerebral hemodynamics and brain tissue oxygenation, while CytOx values represent related effects on intracellular oxidative metabolism.

The purpose of this prospective clinical study was to evaluate new resorbable implants for bone fixation. The plates and screws are made of poly (D, L)lactide (PDLLA). Bioresorbable osteosynthesis and fixation (ResorbX) has been applied in 22 patients. Indications for operations were craniofacial trauma (malar, orbital-floor or frontal bone fractures) or orthognathic procedures (Le Fort-I-osteotomies) as well as the surgical correction in case of craniofacial syndromes. In the initial follow up, the first patients showed clinically and radiologically uneventful fixation and healing of the bone. There were no implant material related complications. Overall, the advantages of PDLLA-implants appear to be their ease of use, radiolucency and resorption, although further experience is needed to determine the longterm benefits of biodegradable implants.

The use of minimally invasive procedures in maxillofacial surgery will require new technologies involving surgical navigation and techniques. The aim of our studies is to improve the efficacy of image-guided navigation in combination with endoscopically assisted techniques for minimally invasive craniomaxillofacial procedures. Prospective evaluation was made of all patients who underwent surgical procedures using image-guided navigation. The most common type of operations performed were endoscopically assisted interventions within the paranasal sinuses, fracture treatment, the resection of bone lesions and further miscellaneous interventions. Our experience to date suggest that image-data based techniques are eminently applicable, providing a feasible alternative to conventional surgical treatment.

Background: Total circulatory arrest in deep hypothermia, which is used in corrective surgery of complex cardiovascular malformations, has been said to cause brain injury. Near-infrared spectroscopy (NIRS) is a new non-invasive method that potentially monitors changes in cerebral oxygenation and tissue oxygen utilisation. The aim of this experimental study in rabbits was to evaluate the change in intravascular and intracellular oxygenation patterns during cooling, deep hypothermic circulatory arrest and rewarming using a commercially available NIRS-cerebral oximeter., Methods: Ten New Zealand White male rabbits (weight, 3.1+/-0.25 kg BW) were included in this study. All animals underwent cardiopulmonary bypass (CPB), cooling to a rectal temperature below 15 degrees C, 60 min of deep hypothermic circulatory arrest (DHCA) followed by reperfusion and rewarming. Cerebral oxyhaemoglobin (HbO2), deoxyhaemoglobin (HHb) and cytochrome oxidase aa3 (CytOxaa3) concentrations were continuously measured during the entire procedure using the Cerebral RedOx Monitor 2020 (Criticon cerebral redox monitor 2020, Johnson & Johnson Medical). Total haemoglobin concentration (tHb) and regional cerebral oxygen saturation (rSO2) values were calculated by integrated algorithm., Results: In all animals an initial increase of oxygenated haemoglobin (HbO2, rSO2) and a fall in deoxygenated haemoglobin (HHb) were found during cooling on bypass. A slight decrease in CytOxaa3 signal was observed in response to initial cooling. Variation in intravascular haemoglobin oxygenation parameters (HbO2, HHb) were related to haemodynamic changes associated with fluid loading, initiation and termination of CPB, bypass flow rate and cooling and rewarming. When the pump flow was stopped all NIRS parameters, except the HHb value, decreased precipitously during the DHCA-period (P<0.01). After reperfusion and rewarming, all haemoglobin oxygen saturation readings returned nearly to pre-CPB levels (P=0.09), but the CytOxaa3 was still significantly lower than the pre-CPB levels (P<0.05)., Conclusion: The change in the NIRS-derived haemoglobin oxygenation parameters may reflect physiological changes in systemic and cerebral haemodynamics. CytOxaa3 values may represent related effects on cellular oxygenation. Thus, continuous, real-time NIRS-monitoring may identify critical periods with inadequate brain tissue oxygenation, particularly during DHCA. The neurological implications of the observed changes in NIRS oxygenation parameters, however, require further quantitative morphological evaluation of the brain in animals surviving a longer reperfusion and observation period.

Pulsewave velocity analysis is an option for the noninvasive determination of cardiac output and the evaluation of additional haemodynamic parameters. An algorithm we developed ourselves has been established as a method for the measurement of cardiac output in rabbits. The effectiveness of this program was investigated by monitoring controlled hypoxia-induced alterations in the circulatory system. Calculated values were compared with direct measurements of cardiac output with a Doppler flow probe placed in the ascending aorta. Within the physiological framework of blood gas analysis, a good correlation was found between the two methods. In the case of hypoxia-induced depression, however, the two methods showed diverging results, presumably due to arrhythmia-induced wave reflections within the arterial vascular tree. This makes an extension of the algorithm necessary to take account for these case.

The aim of this study was to evaluate the relationship between the kinetic patterns of the protein S-100beta, an astroglial cell marker, and its immunohistochemical expression in the brain in rabbits that underwent cardiopulmonary bypass with deep hypothermic circulatory arrest. Fourteen New Zealand rabbits (weight, 3.1+/-0.25 kg) were anaesthetised, intubated and mechanically ventilated. Four animals were not connected to the cardiopulmonary bypass and served as controls. Ten animals were perfused according to a uniform protocol. After systemic cooling, deep hypothermic circulatory arrest was induced for 60 minutes. After reperfusion and rewarming, the animals were weaned from bypass and sacrificed. In the brain, astrocyte reactivity for S-100beta was evaluated immunocytochemically (DPC Immustain) and the serum concentrations of S-100beta were analysed using a commercially available immunoluminometric kit (Byk-Sangtec, Dietzenbach, Germany). In all experimental animals a significant increase of the serum concentration of the protein S-100beta was found immediately after reperfusion and the termination of cardiopulmonary bypass. In comparison with the control animals, increased staining of S-100beta was found in the astroglial cells and swollen astrocytic end-feet in the perivascular regions. There were fewer signs of neuronal cell injury of neurones in the hippocampus structure. In conclusion, astrocytic activation and S-100beta overexpression seems to precede the neurodegeneration following deep hypothermic circulatory arrest. The marked perivascular cell swelling may support the assumption of reperfusion injury of the astroglial cell complex that forms the blood-brain barrier, which may be indicative of the source of the released S-100beta into the blood stream.

Using a dual channel near infrared (NIR) in vivo optical spectroscopy (INVOS) system (INVOS 3100A, Somanetics Corp. Troy, MI, USA) we investigated the relationship between jugular venous oxygen saturation (SjvO2) and regional cerebral oxygen saturation (rSO2) in 30 infants and children (mean age 4.5 years) with congenital heart disease undergoing cardiac catheterisation. The NIRS-SomaSensor (emitter and dual receiver probe) was applied at a standardised right fronto-temporal location (over the right frontal cortex) on the infant's head and covered with an adhesive flexible bandage. Using NIR light (730 and 810 nm) and two source-detector spacings (3 and 4 cm from the transmitter), percentage values of rSO2 were calculated from detected haemoglobin saturations. Simultaneously, jugular venous oxygen saturation (SjvO2) monitoring was performed via a catheter placed in the right internal jugular vein with its tip positioned in the jugular bulb, as verified by fluoroscopy. To compare the reliability of NIRS measurement characteristics, jugular venous blood was analysed for SjvO2 as a reference measure of global cerebral oxygenation, by co-oximetry (OSM3-Hemoximeter, Radiometer Copenhagen, Denmark). Other measured variables included pulse oximetry, arterial blood pressure, and venous and arterial oxygen saturations. Over a jugular venous oxygen saturation range of 31-83%, a significant positive linear correlation was found between rSO2 (NIRS measurement) and SjvO2 (jugular bulb oximetry) (r = 0.93, p < 0.001). No significant correlation was observed between rSO2 values and arterial blood saturation or pulse oximetry. The quantitative correlation between rSO2 (haemoglobin oxygenation in a small hemi-elliptical area of the brain) and reference SjvO2 measurement (method for monitoring global cerebral oxygenation) suggests that NIRS measurement with subtraction algorithm should identify predominantly intracranial saturation in the pediatric age group, and will tend to reflect global oxygenation under physiological conditions. Transcranial oximetry using dual receiving channel NIRS offers a noninvasive, real-time, reliable and practicable means of monitoring cerebral haemoglobin oxygenation changes infants and children with cyanotic and noncyanotic congenital heart disease.

Objective: Recent observations have been shown that the induction and accumulation of heat shock proteins (HSPs) by short exposure to nonlethal whole-body hyperthermia with normothermic recovery are closely associated with transient resistance to subsequent ischemia-reperfusion challanges. Here, this study was performed to investigate whether a shortly heat shock pretreatment affects the left ventricular (LV) function after cold cardioplegic ischemia in reperfused neonatal rabbit hearts., Methods: Hearts from neonatal New Zealand White rabbits were isolated perfused (working heart preparation) and exposed to 2 h of cold cardioplegic ischemia followed by reperfusion for 60 min. To induce the heat shock response neonatal rabbits (n=5, HT-group) were subjected to whole-body hyperthermia at 42.0-42.5 degrees C for 15 min, followed by a normothermic recovery period of 60 min, before harvesting and the onset of global hypothermic cardioplegic arrest. Another set of hearts (n=5, control group) without a heat treatment underwent a similar perfusion and ischemia protocol served as control. The postischemic recovery was assessed by measuring several parameters of LV function. LV biopsies from all control and heat treated animals were taken before ischemia and at the end of reperfusion to examine myocardial HSP levels by Western blot analysis., Results: At 60 min of reperfusion the HT-group showed significant better recovery of ventricular function such as LV developed pressure (DP) (74.6+/-10 vs. 52.1+/-8.5%, P<0.05), LV positive dP/dt (910+/-170 vs. 530+/-58 mmHg/s, P<0.01) and LV end-diastolic pressure (LVEDP) (8+/-2 vs. 18.4+/-5 mmHg, P<0.05) than control. Myocardial oxygen consumption (MVO(2)) was significantly higher in the HT-group compared with control (0.054+/-0.006 vs. 0.041+/-0.002 ml/g per min, P<0.05). Significant postreperfusion lower level in lactate production was observed in the HT-group (0.83+/-0.11 vs. 1.67+/-0.8 mmol/l, P<0.05). Also, the recovery of hemodynamic parameters such as aortic flow, coronary flow and cardiac output was significantly superior (P<0.05) in the HT-group. Furthermore, high expression of HSP72(+)/73(+) were detected in the myocardial tissue samples of heat-treated rabbits by immunoblotting, appearing even at 60 min of normothermic recovery after heat stress., Conclusions: These data in the immature rabbit heart indicate that previous shortly heat treatment with high level expression of heat shock proteins (HSP72(+)/73(+)) before hypothermic cardioplegic ischemia provides transient tolerance against myocardial injury and could be an improvement for the postischemic functional recovery of neonatal hearts.

Currently, no reliable minimally invasive method of measuring cardiac output continuously in neonates and children undergoing cardiac surgery is available. An extravascular Doppler probe was used to measure cardiac output in 15 New Zealand White rabbits (average weight 3.5 kg, range 2.5-4.5 kg). The results obtained were compared with cardiac outputs determined using the aortic thermodilution principle. The mean cardiac outputs measured with the extravascular Doppler probe was 0.37 +/- 0.01 l/min as compared with 0.39 +/- 0.01 l/min with aortic thermodilution. Regression analysis revealed a close correlation (r = 0.973) between the two techniques. The extravascular Doppler techniques is an option for continuous and reliable cardiac output measurement in small animals used in surgical experiments (open chest models) and in neonates or children during surgical repair of complicated congenital heart conditions.

Early detection of myocardial ischaemia following cardiac surgery remains a central problem of intensive care medicine. On the basis of the observation in the epicardium that an elevated potassium activity accompanies myocardial ischaemia, a miniaturized electrode system for the measurement of epicardial potassium concentration was constructed and tested. Different types of electrode were implanted in two groups of rabbits. The electrode parameters were satisfactory (slope, screening) and it proved possible to record interference-free myocardial ion potentials, which with regard to the ionophores used is selective for potassium ion activity in the solid phase electrode. The system thus meets the prerequisites for further investigations into its suitability for early diagnosis of ischaemia.

Introduction: Ischemic periods very often occur a dysfunction of the heart. In this series the intraoperative changes of potassium-activity and pH- value on the heart undergoing cardioplegic induced cardiac arrest were studied. Furthermore, the sensitivity of the system in diagnostic fields of ischemic periods and reperfusion damages were investigated., Methods: In eight patients (7 male and 1 female pts.) undergoing elective cardiosurgical procedures (aortocoronary bypass grafts) the ion activity of potassium and hydrogen ions were monitored on the heart, intraoperatively. By ion-selective electrodes the superficial cardiac ion activities were evaluated on definitive time periods on the left (LV) and right ventricle (RV)., Results: Preliminary results suggest: In the reperfusion-period different pH-values result on the ventricles. In the end of reperfusion the pH-values return to the starting points. Though the evaluated parameters are too alcaline in comparison to physiological intervals. The time course of epicardial potassium-concentrations is not different between the RV and LV. Initially, only low concentrations were found during perfusion and reperfusion, in the last interval of reperfusion period an increase of potassium on both ventricles were evaluated. In the end of the operations the superficial ion concentrations approximate to the values evaluated at the starting points., Conclusion: These results suggests, that the dynamics of ion-exchange-processes in the myocardium can be monitored with the help of ion-selective probes. This new technique opens new strategies in the qualitative check-up of myocardial protective procedures during cardioplegic cardiac arrest.

In this study, the possibility of total intravenous anaesthesia with the anaesthetic agents etomidat/fentanyl, ketamine/xylazine and maintenance with halothane, nitrous oxide and oxygen was used in rabbits for long-term experimental surgery with median sternotomy. The clinical course, cardiovascular and respiratory parameters undergoing long-term anaesthesia for experimental cardiac surgery in rabbits were evaluated.

The study was done in order to investigate the central nervous activities in rabbits (n = 10) during experimental surgery in ketamine and ketamine/midazolam-anaesthesia. The combination of a benzodiazepine with ketamine was thought to be beneficial to reduce the usual excitatory nervous effects after ket amine-anaesthesia. The results of the measured EEG-power spectra and somatosensory-evoked potentials were presented. Additionally, the central effects were correlated with mean arterial blood pressure and the results of blood gas analyzes.