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The aim of the study was to evaluate the effects of long-term confinement on stress-permissive neuroendocrine and immune responses in humans. Two groups of four male subjects were confined 240 days ( group 240) or 110 days ( group 110) in two space modules of 100 or 200 m3, respectively. During confinement, none of the volunteers developed psychic stress as could be examined and verified by a current stress test. However, in g roup 240 but not in group 110, the diurnal rhythm of cortisol secretion was slightly depressed and the urine excretion of norepinephrine significantly increased. The innate part of the immune system became activated as seen by a rise in the number of circulating granulocytes and the enhanced expression of β2-integrins. In contrast, the ratio of T-helper to T-suppressor cells decreased. All these effects, observed during confinement, were even more pronounced in both groups when values of endocrinological and immunological parameters were compared between before and 1 wk after the end of the confinement period. Hence, return to normal life exerts pronounced effects to a much higher degree, irrespective of how long or under which conditions individuals were confined. Because the delayed-type hypersensitivity skin reaction against recall antigens remained unaffected, it is to be presumed that confinement appears to induce distinct sympathoadrenergic activation and immunological changes but no clinically relevant immunosuppression.

We used venous congestion strain gauge plethysmography (VCP) to measure the changes in fluid filtration capacity ( K f), isovolumetric venous pressure (Pvi), and blood flow in six volunteers before, on the 118th day (D118) of head-down tilt (HDT), and 2 days after remobilization (Post). We hypothesized that 120 days of HDT cause significant micro- and macrovascular changes. We observed a significant increase in K f from 3.6 ± 0.4 × 10−3 to 5.7 ± 0.9 × 10−3ml · min−1 · 100 ml−1 · mmHg−1 (+51.4%; P < 0.003), which returned to pretilt values (4.0 + 0.4 × 10−3ml · min−1 · 100 ml−1 · mmHg−1) after remobilization. Similarly, Pvi increased from 13.4 ± 2.1 mmHg to 28.9 ± 2.8 mmHg (+105.8%; P < 0.001) at D118 and was not significantly different at Post (12.4 ± 2.6 mmHg). Blood flow decreased significantly from 2.3 ± 0.3 to 1.3 ± 0.2 ml · min−1 · 100 ml tissue−1 at D118 and was found elevated to 3.4 ± 0.7 ml · min−1 · 100 ml tissue−1at Post. We believe that the increased K f is caused by a higher microvascular water permeability. Because this may result in edema formation, it could contribute to the alterations in fluid homeostasis after exposure to microgravity.

Because 6° head-down tilt (HDT) is an established method to mimic low gravity on earth, the aim of the present study was to determine the effects of 120-day HDT on psychic stress and peripheral blood immune cells in six healthy male volunteers. Psychological state was assessed by a current stress test, and cortisol was measured in saliva. During HDT, all volunteers developed psychic stress, and the diurnal rhythm of cortisol secretion was significantly altered. In addition, urine excretion of dopamine and norepinephrine increased. The innate part of the immune response was activated, as evidenced by the increase in the expression of β2-integrins on polymorphonuclear leukocytes and a rise in the number of circulating natural killer (NK) cell lymphocytes. The ratio of T-helper to T-cytotoxic and T-suppressor cells decreased, whereas no changes in T and B lymphocytes were observed. Plasma levels of interleukin-6 increased significantly and returned to basal levels after the end of the HDT period. Thus 6° HDT appears to be a valid model to induce psychic stress and neuroendocrine-related changes in the immune system, changes that might also be encountered by astronauts and cosmonauts during long-duration spaceflights.

Sixteen patients (1-8 years) scheduled for major general surgery were chosen for the study. They were divided into two groups according to the replacement solution used for haemodilution (HD); whether 6% middle molecular weight hydroxyethyl starch (HES) or 6% dextran 60 (DEX). After induction of general anaesthesia and pulmonary artery catheterization, a precalculated amount of autologous blood was withdrawn while the patient's autologous blood was simultaneously replaced by either HES or DEX. Autologous blood was retransfused at a minimum haematocrit (Hct.) of 17% or at the end of surgery. The following parameters were measured and/or calculated before and after HD, every 20 min intraoperatively and hourly for 6 h postoperatively: heart rate (HR), mean arterial pressure (MAP), Cardiac index (CI), Hct., arterial and mixed venous oxygen content (CaO2, CvO2) and arterio-venous difference of oxygen content (avDO2), oxygen delivery index (DO2I), oxygen consumption index (VO2I). The cardiovascular system remained stable. There was no significant difference as regards SvO2, despite a significant decrease in CaO2 to 10.8 and 10.0 ml.dl-1 (median values) due to reduction of haemoglobin concentration in the HES and DEX groups respectively. In spite of the low hct. values during surgery DO2I remained in normal range (median value 602 and 710 ml.min-1.m-2) in HEX and DEX group respectively. There was no significant change in VO2I after haemodilution (median value 212 and 243 ml.min-1.m-2) in either group. No statistically significant difference was noticed between either groups regarding: CaO2, CvO2, DO2I, VO2I, and no side effects of the colloids were observed. Isovolaemic haemodilution (Hct. approximately 17%) is well tolerated by young children undergoing major elective surgery; global tissue oxygenation was preserved throughout the procedure and both solutions used for haemodilution were equally effective.

A largely positive perioperative fluid balance during both elective and emergency abdominal aortic aneurysm repair (AAA) may put patients at risk of developing left ventricular failure and may thus contribute to morbidity. In the present paper we report on a prospective study using hyperosmotic-hyperonocotic solutions (HHS) infused during clamping of the aorta, for the prevention of declamping shock, and the associated reduction in perioperative fluid requirements. The major aim of this paper was to determine the efficacy of an HHS infusion when given over 20 minutes and to detect possible adverse effects of HHS. For perioperative fluid replacement 12 patients received crystalloid solutions with HHS [250 ml of 7.2% NaCl combined with either 6% Dextran (n = 3), 6% Hydroxyethylstarch (HES, n = 4) or 10% HES (n = 5)]. In 16 controls, crystalloids with 1000 ml of HES 10% were infused. Patients were invasively monitored and hemodynamic parameters frequently assessed during the operation, which were statistically analyzed in relation to the start of the fluid loading during clamping of the aorta. One patient showed an anaphylactoid reaction to HES, otherwise no side effects of HHS were observed during infusion (no hypotension, no pathological EKG changes). Plasma sodium and chloride concentration as well as osmolality rose resulting in an osmotic gradient and a desired intravascular volume expansion. Prior to declamping pulmonary capillary wedge pressure had increased to the desired value of > 13 mmHg and < 18 mmHg. Oxygen delivery was significantly elevated upon HHS and remained so post declamping, whereas no change was observed in controls. During clamping systemic vascular resistance was significantly decreased, but was unchanged in controls. The perioperative fluid balance of patients receiving HHS was 2471.0 +/- 948.6 ml, which was significantly less than + 3386.7 +/- 1247.9 ml of controls (P < 0.01). We suggest that HHS opens new perspectives in perioperative fluid management of both elective and emergency AAA repair, since hemodynamic parameters are improved and the overall fluid balance is less positive, thus decreasing the likelihood of edema formation. Moreover, the previously described positive microcirculatory effects of HHS may be particular beneficial in some high-risk patients.

In order to investigate laser Doppler (LD) flux motion in healthy subjects and patients with peripheral arterial occlusive disease (PAOD), spectrum analysis of LD signals is needed. Autoregressive analysis (AR) is presented as an alternative method of power spectrum estimation. This procedure is compared to the commonly used fast Fourier transform algorithm (FFT) by describing the analytical power of both spectra in the analysis of flux motion waves. LD signals were recorded from the forefoot of 8 healthy volunteers and 11 patients with different degrees of PAOD. The flux, concentration of moving blood cells and velocity signal was digitized and stored for off-line analysis. Special software was designed to calculate AR and FFT spectra of the LD signals and to compare the suitability of both methods for the spectral analysis of LD recordings. Additionally, three-dimensional spectrum diagrams were calculated to demonstrate time-dependent flux changes during standardized provocation maneuvers. AR facilitates the determination of frequency and amplitude of flux motion waves as compared to the FFT. Low frequency-large amplitude waves (LF waves) were detected in both groups. High frequency-small amplitude waves (HF waves), which predominantly appear in severe ischemia, were observed in 7 of the 11 patients and in 2 of the 8 controls. The spectra revealed pulsatile waves in all healthy controls, but only in 1 of the 11 patients. AR modelling allows a reliable description of important flux motion components and has considerable advantages in spectral estimation of LD signals as compared to the FFT.

Nitric oxide (NO) is a pathophysiological mediator with unique signal transducing properties. Signaling mechanisms are categorized as cGMP-dependent or cGMP-independent. Multiple interactions of NO with oxygen, superoxide, and transition metals determine the biological activity. Cyclic GMP-independent responses of NO account for the antimicrobial, the cytostatic, and the cytotoxic capacity of NO. Cytotoxicity is not only directed to harmful cells but also affects the NO-producing cell in a self-destructing loop. For macrophages and pancreatic beta-cells (RINm5F), we established NO-mediated apoptotic cell death. Endogenously generated or exogenously applied NO causes DNA cleavage after endonuclease activation. NO-mediated accumulation of the tumor suppressor p53 precedes apoptotic cell death.

Background/Aims: Degradation of adenine nucleotides to adenosine has been suggested to play a critical role in ischemic preconditioning (IPC). Thus, we questioned in patients undergoing partial hepatectomy whether (i) IPC will increase plasma purine catabolites and whether (ii) formation of purines in response to vascular clamping (Pringle maneuver) can be attenuated by prior IPC. Methods: 75 patients were randomly assigned to three groups: group I underwent hepatectomy without vascular clamping; group II was subjected to the Pringle maneuver during resection, and group III was preconditioned (10 min ischemia and 10 min reperfusion) prior to the Pringle maneuver for resection. Central, portal venous and arterial plasma concentrations of adenosine, inosine, hypoxanthine and xanthine were determined by high-performance liquid chromatography. Results: Duration of the Pringle maneuver did not differ between patients with or without IPC. Surgery without vascular clamping had only a minor effect on plasma purine concentrations. After IPC, plasma concentrations of purines transiently increased. After the Pringle maneuver alone, purine plasma concentrations were most increased. This strong rise in plasma purines caused by the Pringle maneuver, however, was significantly attenuated by IPC. When portal venous minus arterial concentration difference was calculated for inosine or hypoxanthine, the respective differences became positive in patients subjected to the Pringle maneuver and were completely prevented by preconditioning. Conclusion: These data demonstrate that (i) IPC increases formation of adenosine, and that (ii) the unwanted degradation of adenine nucleotides to purines caused by the Pringle maneuver can be attenuated by IPC. Because IPC also induces a decrease of portal venous minus arterial purine plasma concentration differences, IPC might possibly decrease disturbances in the energy metabolism in the intestine as well.

In the pulmonary circulation, endotoxemia induces leukocyte/endothelium interaction in pulmonary arterioles and venules. Thus, leukocytes may contribute to the pathogenesis of acute lung injury. In order to investigate, whether this interaction can be inhibited by early blockade of the adhesion cascade, we studied the effect of the humanized anti-L-selectin antibody HuDreg 200 on leukocyte kinetics in pulmonary arterioles and venules following i.v.-infusion of endotoxin. In endotoxemia HuDreg 200 reduces sticking of leukocytes in both pulmonary arterioles and venules. Thus, we were able to demonstrate that early blockade of the adhesion cascade effectively prevents leukocyte accumulation in pulmonary microvessels in endotoxemia. Therefore, HuDreg 200 may exhibit protective effects on the manifestation of leukocyte-mediated acute lung injury.

Adjunct diagnostic techniques might help surgeons to accurately analyse the depth of a burn. However, despite all technical innovations an ideal device for such an application has not been established for routine use as yet. OPS imaging implemented into the CYTOSCAN A/R is a new, recently introduced technique which allows to obtain high contrast images of the microcirculation without the necessity for fluorescent dyes. The aim of the study was to validate OPS imaging as a tool to study microcirculation in skin after burn injury. OPS imaging was applied by no-touch technique and capillary blood flow was videotaped. Subsequent measurements of the microcirculation were performed at the identical site of the burn. Quantitative analysis of the microcirculation was performed off-line using CapImage. OPS imaging produces high quality images of the microcirculation in a burn wound. Data is given as the number of perfused capillaries per observation area (functional capillary density; FCD) [n/cm2]. OPS imaging allows for direct in vivo visualization and quantification of the microcirculation in burned skin. Our preliminary results of the use of OPS imaging in assessing the microcirculation in burns appear promising, and we hope that this novel technique will allow to improve the knowledge of the dynamics of the microcirculation in the pathophysiology of thermal injury.

The concept of small-volume resuscitation (SVR) using hypertonic solutions encompasses the rapid infusion of a small dose (4 ml per kg body weight, i.e. approximately 250 ml in an adult patient) of 7.2-7.5% NaCl/colloid solution. Originally, SVR was aimed for initial therapy of severe hypovolemia and shock associated with trauma.The present review focuses on the findings concerning the working mechanisms responsible for the rapid onset of the circulatory effect, the impact of the colloid component on microcirculatory resuscitation, and describes the indications for its application in the preclinical scenario as well as perioperatively and in intensive care medicine.With respect to the actual data base of clinical trials SVR seems to be superior to conventional volume therapy with regard to faster normalization of microvascular perfusion during shock phases and early resumption of organ function. Particularly patients with head trauma in association with systemic hypotension appear to benefit. Besides, potential indications for this concept include cardiac and cardiovascular surgery (attenuation of reperfusion injury during declamping phase) and burn injury. The review also describes disadvantages and potential adverse effects of SVR:Small-volume resuscitation by means of hypertonic NaCl/colloid solutions stands for one of the most innovative concepts for primary resuscitation from trauma and shock established in the past decade. Today the spectrum of potential indications involves not only prehospital trauma care, but also perioperative and intensive care therapy.

Hypertonic saline prevents vascular adherence of neutrophils and ameliorates ischemic tissue injury. We hypothesized that hypertonic saline attenuates N-formyl-methionyl-leucyl-phenylalanine (fMLP)-stimulated expression of adhesion molecules on human polymorphonuclear leukocytes (PMNLs). fMLP-stimulated up-regulation of beta2-integrins was diminished by hypertonic saline but not by hypertonic choline chloride-, mannitol-, or sucrose-modified Hanks' buffered salt solution. Shedding of L-selectin was decreased by hypertonic saline and choline chloride but not by hypertonic mannitol or sucrose. When the effects of hypertonic sodium chloride- and choline chloride-modified media were compared, neither solution affected fMLP-receptor binding but both equally inhibited fMLP-stimulated increase in intracellular calcium, ionophore A23187, and phorbol myristate acetate (PMA)-stimulated numerical up-regulation of beta2-integrins. Analysis of mitogen-activated protein (MAP) kinases p38 and p44/42 for phosphorylation revealed that hypertonic solutions did not differ in preventing fMLP-stimulated increases in phospho-p38 and phospho-p44/42. Resting PMNLs shrunk by hypertonic saline increased their volume during incubation and further during chemotactic stimulation. Addition of amiloride further enhanced inhibition of up-regulation of beta2-integrins. No fMLP-stimulated volume changes occurred in PMNLs exposed to hypertonic choline chloride, resulting in significant cell shrinkage. Results suggest a sodium-specific inhibitory effect on up-regulation of beta2-integrins of fMLP-stimulated PMNLs, which is unlikely to be caused by alterations of fMLP receptor binding, decrease in cytosolic calcium, attenuation of calcium or protein kinase C-dependent pathways, suppression of p38- or p44/42 MAP kinase-dependent pathways, or cellular ability to increase or decrease volumes.

We report on the use of OPS imaging to visualize the microcirculation in a burn wound. This novel technique produces high quality images of the microcirculation and quantitative analysis of functional capillary density (FCD) was possible. FCD measured at day 3 following the injury was 11.2 +/- 4.6 (n/mm superset2; mean +/- SEM). During the initial phase of healing microcirculatory changes were characterized by a moderate but steady increase of FCD, which showed marked increase beginning from day 12 following the burn (16.6 +/- 6.9). Maximal FCD measured at day 23 (48.2 +/- 19.7) decreased from this point in time to finally reach 25.2 +/- 10.3 n/mm superset2 at the end of observation. OPS imaging allows for direct in vivo visualization and quantification of the microcirculation in burned skin. Our results of the use of OPS imaging in assessing the microcirculation in a burn wound appear promising, and we hope that this novel technique will allow to improve the knowledge of the dynamics of the microcirculation in the pathophysiology of thermal injury.

Exposure of human mammary carcinoma cell line MCF-7 to TNF-alpha leads to apoptotic cell death within 24 h. In search for apoptosis-preventing signals, we identified glucocorticoids as potent death-preventing compounds. Ten nM dexamethasone provided a significant protective effect whereas 100 nM dexamethasone roughly blocked 80 - 90% of TNF-alpha-induced apoptosis. Surprisingly, dexamethasone exerted a protective effect even when supplied several hours after TNF-alpha. This points to a powerful inhibition of even advanced apoptotic processes by dexamethasone. To further pinpoint the anti-apoptotic glucocorticoid action, we investigated the expression levels of several members of the inhibitors of apoptosis (IAPs) family of proteins in response to TNF-alpha and dexamethasone. IAP proteins directly block caspase protease activities including caspase-3, caspase-7, and caspase-9. Exposure of MCF-7 cells to TNF caused an extensive downregulation of cIAP1, cIAP2, and XIAP protein levels. The decline of the IAP protein levels temporally paralleled the appearance of apoptotic DNA fragments which started 12 - 14 h following TNF-alpha addition and maximal effects were seen within 24 h. Coincubation of cells with TNF-alpha and dexamethasone potently blocked cIAP1, cIAP2, and XIAP downregulation. TNF-alpha-mediated IAP protein downregulation was not affected by proteasome inhibitors like lactacystin, ALLN or ALLM, whereas it was blocked by the broad-spectrum caspase inhibitor Z-VAD-fmk which also prevented TNF-alpha-induced apoptotic cell death. These data suggest that inhibition of IAP downregulation mediated by a caspase proteolytic activity constitutes the anti-apoptotic action of glucocorticoids in MCF-7 carcinoma cells.

Perfusion of isolated organs is a common experimental approach. However, the surfaces of the perfusion system might alter the components of the blood and thereby negatively affect organ function. The aim of this study was to minimize the influence of the perfusion system on the blood components and to evaluate the system. Pressure and flow in the perfusion system consisting of a roller-pump, reservoir, oxygenator, hemo-filter and bubble-trap with a total tubing length of 4.5 m are controlled by a computer software (DASYLAB, Datalog, Moenchengladbach, Germany) via a transducer connected to the system. The organ to be perfused is positioned under a microscope (Orthoplan, Leica, Bensheim, Germany), allowing the investigation of microcirculatory parameters. The images raised are recorded on video tapes. To evaluate the system it was perfused with human blood (Hct 28 to 30%) for 90 min. Heparin (n = 6) or citrate (n = 6) served as anti-coagulants. The disappearance of cells from the blood was determined at time points 0, 1, 5, 10, 15, 20, 30, 45, 60, 75 and 90 min by means of a cell counter (AC T8, Coulter Beckmann, Krefeld, Germany). Cell activation was assessed by analysis of the expression of L- and P-selectin and CD11b. The activation of the complement system was examined by measuring the serum levels of the complement factors C3c and C4. There was no significant loss or activation of the blood cells at any of the above given time points. The serum levels of the complement factors remained within the physiological range and showed no changes throughout the whole experiments. Thus, the perfusion system does not have a negative influence on the blood and its individual components, and is therefore a reliable tool for perfusion experiments.

Apoptosis is a physiological mechanism for the removal of unwanted or damaged cells. Apoptotic cells are rarely seen in living tissues, however, because of their rapid and efficient removal by phagocytosis. Phagocytotic cells such as macrophages or dendritic cells recognize apoptotic cells by specific changes of cell surface markers, which usually are not present on normal cells. One such event is the exposure of phosphatidylserine, which moves from the plasma membrane inner leaflet to the outer leaflet in preapoptotic cells. An unresolved problem, however, was the nature of the phosphatidylserine receptor on the phagocytotic cells. In a recent issue of Nature, Fadok et al. have reported the cloning of a phosphatidylserine receptor using an antibody raised against activated macrophages. Antibody treatment of these macrophages blocks this capacity to engulf.

Fluorocarbon emulsions have been proposed as temporary artificial oxygen carriers. The aim of the present study is to compare the effectiveness of perflubron emulsion with the effectiveness of autologous blood or colloid infusion for reversal of physiologic transfusion triggers.A multinational, multicenter, randomized, controlled, single-blind, parallel group study was performed in 147 orthopedic patients. Patients underwent acute normovolemic hemodilution with colloid to a target hemoglobin of 9 g/dl with an inspiratory oxygen fraction (FIO2) of 0.40. Patients were then randomized into one of four treatment groups after having reached any of the protocol-defined transfusion triggers including tachycardia (heart rate125% of posthemodilution rate or110 bpm), hypotension (mean arterial pressure75% of posthemodilution level oror = 60 mmHg), elevated cardiac output (150% of posthemodilution level) or decreased mixed venous oxygen partial pressure (PVO2;38 mmHg). Treatments in the four groups were 450 ml autologous blood harvested during acute normovolemic hemodilution given at FO2 = 0.40; 450 ml colloid at FIO2 = 1.0; 0.9 g/kg perflubron emulsion with colloid (total = 450 ml) at FIO2 = 1.0; and 1.8 g/kg perflubron emulsion with colloid (total = 450 ml) at FIO2 = 1.0. The primary endpoint was duration of transfusion-trigger reversal. A secondary end-point was percentage of transfusion-trigger reversal.Perflubron emulsion was well tolerated with no serious adverse event attributed to drug treatment. Duration of reversal was longest in the 1.8 g/kg perflubron group (median, 80 min; 95% confidence interval, 60-100 min; P = 0.014 vs. autologous blood, P0.001 vs. colloid) followed by the 0.9 g/kg perflubron group (median, 59 min; 95% confidence interval, 40-90 min), the autologous blood group (median, 55 min; 95% confidence interval, 30-70 min) and the colloid group (median, 30 min; 95% confidence interval, 27-60 min). Percentage of reversal was also highest in the 1.8 g/kg perflubron group (97%; P0.001 vs. autologous blood; P = 0.014 vs. colloid), followed by 0.9 g/kg perflubron (82%), colloid (76%), and autologous blood (60%).Perflubron emulsion (1.8 g/kg) combined with 100% oxygen ventilation is more effective than autologous blood or colloid infusion in reversing physiologic transfusion triggers.

Orthostatic dysregulation is a frequent phenomenon in pilots experiencing extreme G forces and after prolonged exposures to microgravity in cosmonauts. We used non-invasive venous congestion plethysmography (VCP) to study microcirculatory changes and the ability to activate peripheral and centrally mediated protective mechanisms during various degrees of tilt, which we used as an orthostatic challenge.The study, which was approved by the local ethical committee of the Institute of Biomedical Problems, was performed on six healthy 20 - 26 years old male volunteers. We applied 6 - 8 cumulative small venous congestion pressure steps (8 mmHg) to the thigh and determined the fluid filtration capacity (Kf), the linear relationship between cuff pressure (Pcuff) and measured fluid filtration (Jv). We then measured the fluid filtration (Jv) response to varying cumulative degrees of tilt, starting at 0 degrees followed by head down -8 degrees -15 degrees, -30 degrees -15 degrees, -8 degrees, 0 degrees, and then head up 15 degrees, 30 degrees, 70 degrees, 30 degrees, 15 degrees and 0 degrees. Each tilt stage was sustained for 15 minutes. The change in hydrostatic load, at the level of the strain gauge, was determined by measuring the difference in vertical height between the right atrium and mid calf at the level of the gauge. Limb arterial blood flow was measured and lung function tests were performed before and after the small cumulative pressure step protocol, as well as at the end of each tilt step.No significant changes in blood pressure (BP) and heart rate (HR) occurred during the cumulative pressure step protocol. However, an increase in HR was observed during the tilt, but only reached significance at 30 degrees and 70 degrees. The mean Kf value measured with small cumulative pressure steps was 3.25 +/- 0.5 (10(-3) ml.100 ml tissue(-1) mmHg(-1) = KfU), which was significantly (p0.005) higher than the value obtained using tilt to increase the hydrostatic load (0.98 +/- 0.2 KfU). One subject had an unchanged Kf value and experienced vaso - vagal syncope following the imposition of 70 degrees tilt, with a heart rate35 / min and a systolic blood pressure of 60 mmHg. The measurement of blood flow in the limb showed no significant change during the tilt procedure. The lung function measurements revealed, that only expiratory reserve volume (ERV) did significantly increase during 35 degrees and 70 degrees head up tilt. All other parameters were unchanged. We suggest that the differences in these Kf values can be explained by the activation of both central and peripheral mediated changes in pre- capillary resistance in response to the tilt. We suspect, that the vaso - vagal syncope, experienced by one subject, reflected his inability to activate these protective reflex mechanisms, a situation that could be exacerbated by sub-clinical venous insufficiency. We believe that these results show that VCP is a useful tool for the determination of intolerance to orthostatic stress.

There is no mammalian life without oxygen. At the same time, oxygen can be toxic. This paper focuses on extrapulmonary manifestations of oxygen toxicity. Hyperbaric and normobaric applications of oxygen are relevant to the anesthesiologist and carry different risks of oxygen toxicity. Normobaric O2 is tolerated for relatively long periods and has no acute sequelae. Chronic exposure to normobaric O2 will increase the concentration of oxygen radicals in the tissues and produce organ damage. This has to be weighed against the risk of withholding possibly life-saving oxygen therapy. High FiO2 can be used to increase plasma dissolved oxygen to offset low hemoglobin based oxygen transport capacity of blood during acute pre- or intraoperative normovolemic hemodilution with the aim of reducing allogeneic transfusion requirements. Administration of hyperbaric oxygen can be followed by a spectrum of neurologic disturbances leading to frank convulsions. The mechanism leading to cerebral seizure activity is not fully understood but is possibly linked to oxygen radical production and the L-Arginine-NO system.

The expected explosion of costs in transfusion medicine due to the shortfall of healthy donors and the more frequent treatment of transfusion-associated complications (chronic hepatitis, cirrhosis, wound infection, tumor recurrence) increases the socio-economic importance of the development of safe and effective synthetic oxygen carriers as an alternative to the transfusion of homologous red blood cells. Currently two types of artificial oxygen carriers are experimentally and clinically investigated for their capacity to ensure adequate tissue oxygenation in the case of severe anemia. In addition to their oxygen transport capacity solutions based on free human or bovine hemoglobin provide vasoconstrictor properties. Their hyperoncotic properties make them particularly attractive for the treatment of severe hemorrhagic shock. Perfluorocarbon (PFC) emulsions allow an increase of the physically dissolved portion of arterial oxygen content. Due to their particulate nature (emulsion droplets) PFC may only be infused in low doses. Otherwise there is risk of overload and malfunction of phagocytic cells of the reticulo-endothelial system. In the case of an intraoperative blood-loss in preoperatively hemodiluted patients, bolus infusion of PFC represents an effective means to avoid immediate retransfusion of autologous blood and allows for further, extreme hemodilution without risking tissue hypoxia.

Patients undergoing major vascular surgery frequently require a substantial intraoperative fluid replacement to assure hemodynamic stability, which is in excess of the expected fluid requirements due to starving, blood and insensible losses. This leads to a positive fluid balance which can not be readily explained.We have used venous congestion plethysmography (VCP) a non-invasive method for measurement of microvascular parameters in limbs to investigate the changes in microvascular permeability (FFK) and the balance of Starling forces of patients undergoing surgery for unilateral femoral artery reconstruction (FEM) under epidural anaesthesia or abdominal aortic aneurysm repair (AAA) under general anaesthesia. The control group consisted of patients scheduled for inguinal hernia repair or hand surgery under general anaesthesia. All patients were measured 24 hours pre-operatively, immediately after the induction of anaesthesia or completion of epidural anaesthesia and on the 1st, 5th and 10th postoperative day. The perioperative patient management followed a standard protocol and all patients with vascular disease were invasively monitored using indwelling arterial lines and central venous catheters. Continuous infusion of Ringers lactate and 6% Dextran 60 was sustained during the induction period. Each patient gave informed consent.Preoperatively we found no significant difference in the mean FFK-values of controls (4.1 +/- 0.4, ml.min-1 100 ml tissue-1 mmHg-1 x 10(-3) = FFKU), the AAA (3.6 +/- 0.3 FFKU) and FEM (4.2 +/- 0.3 FFKU). After induction of anaesthesia the mean FFK value in the controls fell to 3.5 +/- 0.5 FFKU (p = 0.07), whereas in the AAA patients we observed a significant increase to 4.7 +/- 0.2 FFKU (p0.005) and after epidural anaesthesia in FEM to 5.5 +/- 0.4 FFKU (p0.001) respectively. Those post anaesthetic FFK values where significantly higher in FEM and AAA than in the controls (p0.02). In AAA we found a significant positive correlation between the increase in FFK and the intraoperative fluid balance (r2 = 0.69, p0.01). No such correlation was found in controls and FEM. The postoperative values of FFK where unchanged in the control group, whereas a further increase was seen in both patient groups with vascular disease, with a maximum in AAA on the 1st postoperative day (to 5.4 +/- 0.4 FFKU mean both legs) and the 5th postoperative day in FEM (to 7.3 +/- 1.7 non-ischemic leg, 7.1 +/- 1.2 ischemic leg FFKU). In both groups normal FFK values where found on the 10th day after the operation.The data presented suggests an increase in extravascular fluid loss in patients undergoing vascular surgery, which becomes evident after the induction of general anaesthesia or completion of epidural anaesthesia. The positive correlation with the intraoperative fluid requirements may partially explain the often reported large intraoperative fluid requirements of patients undergoing AAA repair. The fact that the maximum change in fluid filtration capacity is found postoperatively may be explained by the additional effect of an ischemia/reperfusion injury in response to both the clamping an declamping of the artery and the increase in arterial blood flow to the limb due to the successful reconstruction of the blood vessel.

After ischemia-reperfusion, polymorphonuclear leukocytes (PMNLs) become activated by inflammatory mediators, adhere to the vascular endothelium via the interaction of specific adhesion molecules, and cause tissue injury by the release of cytotoxic oxygen radicals and enzymes. Results obtained in animal experiments suggest a key role for PMNLs in ischemia-reperfusion injury of transplanted livers; therefore, we studied the expression of adhesion molecules on circulating PMNLs (beta2-integrins [CD18] and L-selectin [CD62L]) in 20 patients undergoing orthotopic liver transplantation (study group). To determine the effects of surgical trauma to the liver in the absence of ischemia and reperfusion, the expression of PMNL adhesion molecules was measured in 10 patients scheduled for elective partial liver resection without hepatic vascular exclusion (control group). Patients were classified as responders or nonresponders based on changes in the expression of adhesion molecules elicited by reperfusion. In the control group, all patients remained nonresponders, showing that surgical trauma of the liver alone does not cause activation of circulating PMNLs. In contrast, 8 of 20 patients in the study group were classified as responders. In responders, postoperative serum liver enzyme activities were significantly higher than in nonresponders, indicating that activation of PMNL was associated with damage to hepatocellular integrity. Because expression of adhesion molecules was already changed during surgery, monitoring of the expression of beta2-integrins and L-selectin on circulating PMNLs during orthotopic liver transplantation might be useful in prediction of early graft dysfunction.

An increased microvascular water permeability has been reported after ischemia/reperfusion both in animal models and in human studies. We studied the changes in fluid filtration capacity (FFK) after ischemia/reperfusion due to tourniquet in patients undergoing arthroscopy of the knee.Healthy male volunteers (n = 24, mean age 46.9 +/- 3.5) were studied prior to, 1 and 6 hours after arthroscopy of the knee, during which a tourniquet was applied to the thigh. FFK, isovolumetric venous pressure (Pvi) and arterial blood flow in the limb was measured in both legs (tourniquet leg and control leg) using computer assisted venous congestion plethysmography. Venous blood samples were obtained from a cubital vein prior to and from the femoral vein 2 mins after deflation of the tourniquet cuff. 12 patients received preoperatively an infusion of 6% Dextran (D) and 12 patients 500 ml of electrolyte solution (VE) an.The mean duration of the tourniquet was (D) 56.0 +/- 6.9 min and (VE) 53.9 +/- 4.2 min which resulted in a significant increase in venous lactate concentration from (D) 1.4 +/- 0.1 mmol.l-1 to 2.7 +/- 0.3 mmol.l-1 and (VE) 1.3 +/- 0.1 mmol.l-1 to 2.7 +/- 0.3 mmol.l-1 (p0.001). A significant decrease in pH from (D) 7.39 +/- 0.01 to 7.32 +/- 0.01 (p0.001) and from (VE) 7.39 +/- 0.01 to 7.32 +/- 0.01 (p0.001) was also seen. Preoperatively no significant differences in the FFK values of the tourniquet leg (D = 5.3 (4.8-10.7) ml. x 10(-3) min-1.100 ml tissue-1 mmHg-1 = FFKU) and the control leg (5.2 (4.7-8.6 FFKU)) were observed. The maximum FFK value in D was seen 1 hour after ischemia/reperfusion in both, the tourniquet leg (7.5 (4.6-14.2 FFKU) and the control leg (7.8 (5.5-9.4 FFKU). In VE however the maximal FFK value were measured 6 hours after ischemia/reperfusion with an increase in the tourniquet leg from 5.2 (4.2-6.2 FFKU) to 8.1 (3.7-10.4 FFKU) and the control leg from 6.1 (3.6-7.0 FFKU) to 7.3 (6.1-8.3 FFKU) (Median (Range). One-way ANOVA). There were no significant differences in the FFK values between D and VE except for a lower Pvi in VE at the third measurement. No significant changes in the arterial blood flow were seen perioperatively as well as between the operated and non-operated leg.A tourniquet of1 hour does impair tissue oxygenation as indicated by the increase in lactate and the decrease in pH. The duration of the tourniquet was however to short to have caused sufficient microvascular damage result in a more pronounced increase in fluid filtration capacity.

The microcirculation is the organ that provides the direct link between blood and tissue, and thereby between the whole organism and the single cell. Modern microcirculation research in experimental surgery is characterized by the use of high-resolution video fluorescence microscopy and quantitative computer-assisted image analysis coupled with the techniques of molecular biology and transgenic or knockout gene technology. These advances should improve knowledge about surgically relevant physiologic and pathophysiologic phenomena at the interface between blood and tissues and help us to understand the initial molecular mechanisms leading to organ dysfunction following inflammation, ischemia-reperfusion, and transplantation.

1. Photoplethysmography is a widely used non invasive technique for the measurement of peripheral oxygen saturation. A more detailed analysis of the volume pulse (VP) can indicate alterations in peripheral vascular tone, due to sympathetic stimulation, stress, pain and temperature. - 2. In six healthy male volunteers we investigated changes in the VP resulting from vasoconstriction and vasodilatation induced by varies degrees of tilt. Subjects were subjected 0 degrees tilt followed by head down -8 degrees -15 degrees, -30 degrees -15 degrees, -8 degrees, 0 degrees, 15 degrees, 30 degrees , 70 degrees, 30 degrees, 15 degrees and 0 degrees. Each tilt stage was sustained for 15 minutes. Both VP - and haemodynamic changes were continuously recorded 30s before and then for 210 s after the imposition of each tilt step. We used a new computer driven soft and hardware for the analysis of the VP. 3. The VP signal was obtained with a sensor emitting 840 nm (NIR) and 640 nm (RED) light into finger tip with a sample rate of 128 Hz. All data was normalised to the initial mean value obtained at 0 degrees tilt. The signal strength parameters amplitude, and area under the curve and the first derivative of the amplitude (flux) as well as time discrete parameters, time of first maximum (Tmax), dicrote wave (Td), volume pulse decrease (Tdec) and fundamental arterial oscillation Tag = Td - Tmax were measured. 4. HR increased significantly during 30 degrees and 70 degrees tilt, but no change in the other hemodynamic parameters was observed. Amplitude, area under the curve and flux of both the Red and the NIR signal increased following head down tilt. A significant decrease of those parameters was found during foot down tilt. 5. No significant changes were found in the time discrete values, neither within each tilt step nor when compared to the initial mean value at 0 degrees tilt. 6. This study reveals that signal strength related parameters such as area under the curve, amplitude and flux reflect changes in vascular tone. Time discrete parameters however did not depict these changes and appear unsuitable for data analysis when using this specific hardware applied in the current study.

We report the case of a 22-year-old woman who underwent two-step scoliosis surgery without allogeneic transfusion, although the intraoperative blood loss (3500 ml) during the first procedure was higher than the calculated blood volume (3250 ml). Preoperatively the patient had donated four units of autologous blood. Intraoperatively blood-saving methods were combined. During the first operation acute normovolemic hemodilution (target hemoglobin 9.0 g/dl) was applied and during the second operation controlled hypotension (systolic blood pressure 80 mmHg). Intraoperative auto-transfusion was used in both procedures. During the first operation severe normovolemic anemia (minimal hemoglobin 3.5 g/dl) was accepted while the patient was ventilated with FiO2 1.0. The hemoglobin concentration was 8.6 g/dl after the first procedure and had increased to 11.6 g/dl 4 weeks after the second procedure. No severe complications occurred during the postoperative phase. This case report shows that also in surgical procedures with extreme blood loss any allogenic transfusion can be avoided by the combination of blood-saving methods, acceptance of low intraoperative transfusion trigger and ventilation with 100% oxygen.

For ethical and socio-economical reasons (cost-explosion in transfusion-medicine, patient's individual destiny), development and consistent application of allogeneic transfusion sparing techniques in surgery is a challenge to anesthesiologists, surgeons and blood-bankers. The combination of different techniques, i.e. autologous predonation, hemodilution, choice of anesthetic regimen, deliberate hypotension, application of antifibrinolytic agents and autotransfusion of intraoperatively saved blood allow for avoidance of allogeneic blood transfusion even in patients presenting important intraoperative hemorrhage. The present article summarizes (1) risks associated with transfusion of allogeneic blood, (2) actually applied pre- and intraoperative techniques to reduce transfusion of allogeneic blood and (3) new concepts (administration of erythropoietin, hyperoxic ventilation and administration of artificial oxygen carries) to further increase the efficacy of autologous predonation and preoperative normovolemic hemodilution.

Regional organ blood flow (RBF) is spatially heterogeneous. Relative dispersion (standard deviation S.D./mean) is often used to assess heterogeneity of RBF. Relative dispersion is a global measure of heterogeneity and is strongly influenced by the tissue sample size making comparisons between research groups inappropriate. Spatial correlation (SC) of blood flow is, on the other hand, averaged local self similarity. Both parameters change oppositely secondary to interventions. Fractal dimension ( D ) is a scale-independent measure of spatial heterogeneity and thus facilitates comparison of data. Programs for calculation of SC and D have not been published. We present two portable computer programs written in C++ for calculating SC and D . The programs were validated with six computer generated data sets of known heterogeneity. The results were in agreement with data from the literature: we conclude that the programs accurately calculate spatial correlation and fractal dimension of 1-, 2-, or 3-dimensional perfusion matrices.

Uptake of naked functional DNA into mammalian cells can be achieved by a number of physical methods. However, for most of these techniques possibilities for therapeutic in vivo applications – especially to solid organs – are often limited. In this report, we describe shock wave permeabilization as a new physical gene transfer method, which can be easily applied, provides great flexibility in the size and sequence of the DNA molecules to be delivered, and which should exhibit an advantageous security profile in vivo. Upon exposure to lithotripter-generated shock waves eukaryotic cells display a temporary increase in membrane permeability. This effect was shown to be caused by cavitation resulting in the transient generation of cell pores which allows the direct transfer of naked plasmid DNA. Shockwave transfection of a variety of cell lines was demonstrated. Since shock waves can be well focused within particular body regions, future applications of extracorporally generated shock waves to tissues simultaneously perfused with DNA solutions might open up the possibility of achieving a regionally enhanced in vivo gene transfer.

Ischemic diseases of heart and brain are the primary causes of mortality in industrialized nations. The ischemic injury with the consecutive reperfusion is responsible for the disturbance of microcirculation with ensuing tissue damage and organ dysfunction. Recent evidence suggests that oxygen-derived free radicals and activated polymorphonuclear leukocytes produced in ischemic tissue are instrumental in the development of ischemic cell injury. In pancreas, ischemia/ reperfusion is proposed as a potentially damaging factor accounting in part for the pathogenesis of acute pancreatitis. Apart from ischemia/reperfusion injury, the kallikrein-kinin system mediates acute inflammation associated with enhanced capillary permeability and accumulation of polymorphonuclear leukocytes, cardinal features of ischemia/reperfusion injury also in acute pancreatitis. Therefore, it seems reasonable to use bradykinin-antagonists to influence postischemic reperfusion injury of the pancreas. In the following, we describe the pathophysiology of ischemia/reperfusion injury with special reference to the pancreatic microcirculation and morphological changes as observed in a model of complete and reversible ischemia. Furthermore, we will discuss the effects of two bradykinin-antagonists (HOE 140 and CP-0597) on functional integrity of the pancreas after ischemia/ reperfusion.

Venous congestion plethysmography enables noninvasive assessment of microvascular filtration capacity (Kf) in limbs. However, increases in fluid filtration might alter the balance of Starling forces: for example, progressive increases in interstitial fluid pressure (Pi) would reduce net fluid flux, thus underestimating Kf. Furthermore, elevation of cuff pressure to values close to diastolic blood pressure, as used in the protocol, may be itself impair tissue perfusion with unknown effects on the microvascular parameters investigated.Pi was measured in healthy volunteers (n = 14) with a modified "Wick in needle" technique during small (8 mm Hg) cumulative increases in venous pressure (0-95 mm Hg). Changes in the hemoglobin (Hb) concentration, oxygenated hemoglobin (HbO2) concentration and oxidized cytochrome aa3 concentration were assessed in the calf using noninvasive near-infrared spectroscopy. Skin red blood cell flux close to the strain gauge was evaluated by laser Doppler fluxmetry.Pi at control was -0.89 +/- 0.8 mm Hg and during elevation of venous pressure remained constant until a cuff pressure of 30 mm Hg was reached. It rose thereafter to 1.57 +/- 1.3 mm Hg (mean +/- SD). Skin red cell flux was significantly reduced when cuff pressure exceeded 30 mm Hg and following cuff deflation, evidence of reactive hyperemia was obtained. Hb concentration increased significantly as a result of venous pressure elevation. No change in either HbO2 or cytochrome aa3 concentration was observed as long as cuff pressure remained under diastolic blood pressure.The small increase in Pi together with an absence of impaired tissue oxygenation during the venous congestion plethysmography protocol described by Gamble et al. supports the contention that this protocol enables accurate assessment of filtration capacity.

Adhesion molecules on polymorphonuclear leukocytes (PMNL) play an important role in nonspecific defense mechanisms directed at invading microorganisms. When local infection, however, cannot be controlled, a systemic inflammatory response syndrome (SIRS) ensues which may progress to septic shock and multiple organ failure, these being major determinants of the patient's outcome. In the present study, the expression of beta 2-integrins and L-selectin on blood PMNL was measured on subsequent days in patients with sepsis (n = 17) and in healthy volunteers (n = 15). beta 2-Integrins and L-selectin molecules were detected by flow cytometry, using the monoclonal antibodies IB4 (anti-CD18) and Dreg200 (anti-CD62L), respectively. Adhesion molecules were determined at baseline immediately after blood collection and also 45 min after incubation of cells in vitro at body temperature to allow for spontaneous regulation. In addition, PMNL were activated by receptor-dependent and receptor-independent stimuli to characterize stimulus-specific adhesion molecule expression. In parallel with the measurement of adhesion molecules, severity of sepsis was assessed by the Elebute score. The results demonstrate significant differences in the basal, spontaneous and stimulus-induced expression of adhesion molecules between healthy volunteers, survivors (n = 11) and nonsurvivors (n = 6). Moreover, when survivors and nonsurvivors with severe sepsis (Elebute score12) were compared, basal expressions of both beta 2-integrins and L-selectin were significantly lower in patients who did not survive. Thus, measurement of adhesion molecules on circulating PMNL may be useful to identify septic patients at high risk for lethal outcome.

Trauma and associated major blood losses in Germany represent the leading cause of mortality in patients up to 45 years of age. The endpoints of prehospital fluid resuscitation in traumatic-hemorrhagic shock are the restitution of intravascular volume und cardiac preload, in order to increase cardiac output and thus provide adequate oxygen delivery to the tissues. The key therapeutic factor to prevent the development of multiple organ failure complicating trauma and shock, however, is the normalization not only of macrohemodynamics (systemic blood pressure. cardiac output), but the restitution of the disturbed microvascular perfusion. In case of major blood loss and exsanguination this can not be achieved by crystalloids and synthetic colloids used in conventional volume therapy. A new concept consists of i.v. bolus infusion of a small volume (4 ml/kg body weight) of a hyperosmolar (7.2-7.5%) NaCl/colloid solution, which is termed "Small-volume Resuscitation". Recently presented data from a cohort analysis of 8 preclinical studies show an increase in survival rate by about 5% when compared to standard of care. In addition, artificial oxygen carrying solutions are currently investigated. by which-through an increase of O2-blood content-oxygen delivery to the tissues might be augmented.

Nitric oxide (NO) generation initiates apoptotic cell death in different experimental systems. In RAW 264.7 macrophages the appearance of typical apoptotic markers is linked to inducible NO synthase induction. Mechanistically, accumulation of tumour suppressor p53 precedes apoptotic DNA fragmentation. With the use of S-nitroglutathione (GSNO) we correlated a dose-dependent p53 up-regulation to DNA fragmentation measured after 4 h and 8 h, respectively. Our studies revealed a linear correlation between the potency of five different NO donors with respect to apoptosis induction and p53 accumulation. Furthermore, we probed for NO-induced apoptosis after stable transfection of RAW 264.7 macrophages with plasmids encoding p53 antisense RNA. Clones with down-regulated p53 levels in response to GSNO exhibited a marked reduction in DNA fragmentation. Expression of the inducible NO synthase in response to lipopolysaccharide and interferon-γcaused apoptosis in RAW 264.7 macrophages and neomycin-vector controls within 24 h. In contrast, p53 antisense RNA-expressing clones appeared highly resistant towards endogenous NO, although inducible NO synthase induction with concomitant nitrite production remained unchanged. For RAW 264.7 macrophages our results established a functional role of the tumour suppressor p53 during NO-induced apoptotic cell death. However, p53 antisense experiments and the use of the p53-negative cell line U937 substantiated p53-independent signalling pathways operative during NO-mediated apoptosis.

A Roundtable of Experts in Surgery Blood Management was held 7 to 9 April 1995, in Vienna, where all the available published and unpublished epoetin alfa clinical data was thoroughly discussed and where the following Consensus Statement was developed to give the clinician some practical guidelines for the administration of epoetin alfa in elective surgery patients.Consensus Statement: For elective surgery patients it is important to facilitate, via blood conservation methods, the elimination or reduction of exposure to allogeneic blood transfusion and its associated risks. Adjunctive use of epoetin alfa therapy in specific patients is one method to accomplish this goal. Patients with small body size, predicted low blood volume ( < 5 1), and/or a hematocrit too low to be enrolled in or to complete their prescribed ABD (autologous blood donation) program especially benefit from epoetin alfa therapy. Generally, epoetin alfa therapy is well tolerated, with an acceptable risk-to-benefit ratio.

This experimental study compared the effects of laparoscopic (n = 31) and open (n = 32) cholecystectomy on gastric intramucosal pH (pHj). For this purpose, pHj was measured tonometrically before induction of anaesthesia, at 30-min intervals during surgery, and 1, 2 and 4 h after operation in otherwise healthy patients undergoing elective cholecystectomy. Additionally, perioperative arterial pH (pHa), arterial carbon dioxide tension (PaCO2), intramucosal carbon dioxide tension, arterial bicarbonate concentration, end-tidal carbon dioxide pressure (PECO2), levels of serum lactate, lactate dehydrogenase (LDH) and γ-glutamyl transferase (GGT), haematocrit and arterial blood pressure were recorded. In the two groups no significantly different changes occurred in pHi, pHa, serum lactate concentration or haematocrit at any of the observation times. PECO2 and PaCO2 were significantly raised during the laparoscopic procedure, whereas levels of LDH and GGT and arterial blood pressure rose during and after open cholecystectomy. In spite of the increased intra-abdominal pressure and the peritoneal carbon dioxide absorption related to the creation of a pneumoperitoneum, no decrease in pHi was detectable during laparoscopic cholecystectomy.