Biomedical subjects

Peter Biberthaler

Publications and source records attributed to Peter Biberthaler.

At least 19 recordsLinked to original sources

PubMed2006

Measurement of S-100B for risk classification of victims sustaining minor head injury--first pilot study in Brazil.

BACKGROUND: Release of the neuronal protein S-100B into the circulation has been suggested as a specific indication of neuronal damage. The hypothesis that S-100B is a useful and cost-effective screening tool for the management of minor head injuries was tested. METHODS: Fifty consecutive patients sustaining isolated minor head injury were prospectively evaluated in the emergency room of a Brazilian hospital by routine cranial computed tomography scan. Venous blood samples (processed to serum) were assayed for S-100B using a newly developed immunoassay test kit. Twenty-one normal healthy individuals served as negative controls. Data are presented as median and 25 to 75 percentiles. RESULTS: Patients reached the emergency room an average of 45 minutes (range: 30-62 minutes) after minor head injury. Six of 50 patients (12%) showed relevant posttraumatic lesions in the initial cranial computed tomography scan and were counted as positive. The median systemic concentration of S-100B in those patients was 0.75 microg/L (range: 0.66-6.5 microg/L), which was significantly different (U-test, P < .05) from the median concentration of 0.26 microg/L (range: 0.12-0.65 microg/L), of patients without posttraumatic lesions as counted by the cranial computed tomography. A sensitivity of 100%, a specificity of 20%, a positive predictive value of 15%, and a negative predictive value of 100% was calculated for the detection of patients suffering from intracranial lesions. CONCLUSIONS: Protein S-100B had a very high sensitivity and negative predictive value and could have an important role in ruling out the need for cranial computed tomography scan after minor head injury. This appears to be of substantial clinical relevance, particularly in countries where trauma incidence is high and medical resources are limited, such as in Brazil.

Biomarkers↗

PubMed2006

Rapid measurement of S-100B serum protein levels by Elecsys S100 immunoassay in patients undergoing carotid artery stenting or endarterectomy.

OBJECTIVES: This study was designed to apply the rapid Elecsys S100 immunoassay for real-time measurement of S100 protein serum levels indicating acute brain damage in patients undergoing carotid artery stenting (CAS) or endarterectomy (CEA). DESIGN AND METHODS: Data of 14 CAS patients were compared to those of 43 CEA and 14 control patients undergoing coronary angiography (CA). S100 serum levels were measured by the full-automatic Elecsys S100 immunoassay and compared to those obtained by the well-established LIA-mat S100 system. RESULTS: In contrast to CAS and CA patients, median S100 serum levels of CEA patients significantly increased to 0.24 ng/mL before declamping, but subsequently returned to baseline. Three CEA patients with neurological deficits showed sustained elevated S100 levels 6 h after extubation. Absolute S100 values were not significantly different between the two methods. Bland-Altman plot analyses displayed a good agreement, mostly indicating slightly smaller values applying the Elecsys S100 system. CONCLUSIONS: The Elecsys S100 system appears to be suitable for rapid real-time detection of neurological deficits in patients undergoing CAS and CEA. Persistent elevations of Elecsys S100 levels during CEA were associated with prolonged neurological disorders, whereas transient increases seem to represent impaired blood-brain barrier integrity without neurological deficits.

Aged↗

PubMed2006

Analysis of NF-kappaB nuclear translocation in PMN-neutrophils of major trauma patients in the early post-traumatic period: a pilot study.

BACKGROUND: Post-traumatic inflammation is connected to polymorphonuclear neutrophil (PMN)-dysfunction characterized by reduced nuclear translocation of NF-kappaB during the post-traumatic period. However, the dynamic of NF-kappaB translocation in PMN of major trauma patients remains unclear. Hence, the aim of this pilot study was to analyze NF-kappaB in PMN from multiply injured patients immediately after trauma. PATIENTS AND METHODS: Blood samples of major trauma patients (ISS > 16) were drawn on admission within 90 minutes after trauma and at 6, 12, 24, 48, and 72 hours after trauma. Neutrophilic NF-kappaB-translocation was analyzed by EMSA and quantified by densitometry as (arbitrary units). In addition, PMN of healthy volunteers were analyzed either in their native state (-control) or after LPS stimulation (+control). RESULTS: Twelve patients (NISS: 34 +/- 10 [mean +/- SEM]) were enrolled. NF-kappaB translocation was significantly increased in trauma patients on admission and after 6 hours. Interestingly, a second activity peak was present after 24 hours. In patients who later died, NF-kappaB activity was significantly elevated initially, to be rapidly diminished after 6 hours, while it increased in the survivors group. After 24 hours NF-kappaB activity increased significantly in the survivors group, to become reduced in both groups at a later time. CONCLUSIONS: Within this pilot study, the dynamic of NF-kappaB translocation in PMN of multiply injured patients immediately after trauma was analyzed for the first time. Enabled by closely matched sequential blood sampling strictly standardized to the traumatic event, an essential biphasic increase of neutrophilic signal transduction could be investigated in the very early post-traumatic period, which preceded the downregulation of the innate immune system.

Adolescent↗

PubMed2006

Intrathecal and systemic concentration of NT-proBNP in patients with severe traumatic brain injury.

Outcome of patients suffering from traumatic brain injury (TBI) depends on the development of secondary brain damage. In this context, recent studies underlined the role of the natriuretic peptides- atrial natriuretic peptide and brain natriuretic peptide (BNP)-in aneurysmatic subarachnoidal hemorrhage (SAH). Especially BNP correlates with intracranial pressure and clinical outcome after SAH. Since its role in TBI remains unclear, the intracranial and systemic concentrations of N-terminal (NT)-proBNP were analyzed in patients suffering from severe TBI. We measured NT-proBNP levels in cerebrospinal fluid (CSF) and serum of 14 patients suffering from severe TBI (GCS 15 mm Hg (n=6), the serum (800+/-150 pg/mL) and CSF levels (55+/-9 pg/mL) of NT-proBNP were significantly increased after 24 h, as compared to patients with ICP 15 mm Hg. Further studies are currently performed to elucidate the physiologic role of NT-proBNP in TBI.

Adult↗

PubMed2006

Serum S-100B concentration provides additional information fot the indication of computed tomography in patients after minor head injury: a prospective multicenter study.

Ninety percent of patients with minor head injury (MHI) who have cranial computed tomography (CCT) under the present clinical decision rules have normal scans. Serum concentrations of the astroglial protein S-100B were recently found to provide useful information, but these studies were too small to provide a statistically safe basis for changing the present rule. We have investigated whether S-100B concentrations in patients with MHI can provide additional information to improve indication of the need for an initial CCT scan. One thousand three hundred nine patients with MHI were enrolled in this prospective, multicenter study. All had a CCT scan to confirm diagnosis in accordance with the present clinical decision rules. S-100B was measured in serum samples obtained upon admission. Data were analyzed using contingency table and receiver operating characteristic curve and compared with those for healthy donors (n = 540) and with those for patients with moderate to severe head injury (n = 55). Of the 1309 patients studied, 93 exhibited trauma-relevant intracerebral lesions on the CCT scan (CCT+). With a cutoff limit of 0.10-microg/L S-100B (95th percentile of values in healthy volunteers), CCT+ patients were identified with a sensitivity level of 99% (95% confidence interval, 96% - 100%) and a specificity level of 30% (95% confidence interval, 29% - 31%). Adding the measurement of S-100B concentration to the clinical decision rules for a CCT scan in patients with MHI could allow a 30% reduction in scans. A prospective study of the clinical value of S-100B measurement in such patients is now under way.

Adult↗

PubMed2006

Significance of Elecsys S100 immunoassay for real-time assessment of traumatic brain damage in multiple trauma patients.

BACKGROUND: The neuroprotein S100 released into the circulation has been suggested as a reliable marker for primary brain damage. However, safe identification of relevant traumatic brain injury (TBI) may possibly be hampered by S100 release from peripheral tissue. The objective of this study was to measure early S100 levels using the Elecsys S100 immunoassay for real-time assessment of severe TBI in multiple trauma. METHODS: Consecutively admitted multiple trauma patients (injury severity score >or=16 points) were stratified according to the results of the initial cerebral computed tomography (CCT) examination. S100 serum levels were determined at admission and at 6, 12, 24, 48 and 72 h after trauma. Data were correlated to creatine phosphokinase (CK) and lactate dehydrogenase (LDH) serum levels. Using receiver operating characteristic (ROC) analysis, the discriminating power of S100 measurement was calculated for the detection of CCT+ findings. RESULTS: Median S100 levels of CCT+ patients (n=9; 37 years) decreased from 3.30 microg/L at admission to 0.41 microg/L 72 h after trauma. They revealed no significant differences to CCT- patients (n=18; 44 years), but remained elevated compared to controls. Median CK and LDH levels correlated with the corresponding S100 levels during the first 24 h after trauma. ROC analysis displayed a maximum area under the curve of only 0.653 at 12 h after trauma. No significant difference was calculated for the differentiation between CCT+ and CCT- patients. CONCLUSIONS: Measurements of S100 serum levels using the Elecsys S100 immunoassay are not reliable for the real-time detection of severe TBI in multiple trauma patients. Due to soft tissue trauma or bone fractures, S100 is mainly released from peripheral sources such as adipocytes or skeletal muscle cells.

Adult↗

PubMed2005

An enzyme-linked immunosorbent assay for human cathepsin X, a potential new inflammatory marker.

The human lysosomal cysteine-type carboxypeptidase cathepsin X is mainly present in monocytes and macrophages and may be released into the circulation due to constitutive and/or regulated secretion by (activated) immune cells. To define its potential diagnostic value as an inflammatory marker, we have developed a highly sensitive and specific sandwich-type immunoassay (ELISA) for cathepsin X permitting both intra- and extracellular detection and quantification. The dynamic range of the cathepsin X ELISA was determined to be 100 (detection limit) to 8000 pg/ml. Reproducibility of both within and between runs yielded coefficients of variation (CVs) of 2.7-3.5% and 6.3-7.3%, respectively. Cross-reactivity with other members (cathepsin B, L) of the thiol-dependent cathepsin family was not observed. The ELISA was used to quantify cathepsin X in leukocytes as well as in plasma of healthy volunteers and patients with multiple trauma. During the first 72 h after trauma, plasma levels of cathepsin X increased significantly, particularly in patients who died during the posttraumatic period. In comparison to the well-known inflammation marker neutrophil elastase, cathepsin X levels predicted survival with a higher significance in the later posttraumatic phase. In conclusion, this report provides the first evidence of cathepsin X immunoreactivity not only in cell lysates but also in plasma samples. We suggest that the newly developed highly reproducible ELISA will be of great value for further evaluation of this protease as a diagnostic and/or prognostic marker in inflammatory diseases.

Animals↗

PubMed2005

Genome-wide monocytic mRNA expression in polytrauma patients for identification of clinical outcome.

Immune activation in multiple trauma is closely linked to the development of multiple organ dysfunction and failure, and consequently, has a profound influence on patient outcome. Although peripheral blood monocytes play a critical role in this immune response, the biological significance of changes in genome-wide expression immediately after traumatic injury have not been explored previously. Thirteen patients presenting with multiple blunt trauma were studied. Peripheral blood monocytes were obtained within 90 min and at 6, 12, 24, 48, and 72 h after trauma. Apparent genome-wide expression was determined with Affymetrix U133A microarrays. Supervised analysis identified 698 probe sets that were differentially expressed in the 13 trauma subjects (P < 0.001) over the 72-h study period. An additional 763 probe sets were differentially expressed in patients who died (n = 3) compared with those who survived (n = 10). The ability of these probe sets to function as a classifier of survival was significantly demonstrated with six prediction models. Using pathway analysis, a network of proinflammatory genes and intracellular signaling pathways leading to c-JUN activation were consistently overexpressed in patients who died. Genome-wide mRNA expression patterns in circulating peripheral blood monocytes from multiple-injured patients can discriminate clinical outcome. The pattern of gene expression in patients who died suggests that in these individuals, there is a reprioritization of gene expression consistent with an early activation of selected genes involved in the initiation and propagation of a proinflammatory response.

Adult↗

PubMed2005

Effect of stress doses of hydrocortisone on S-100B vs. interleukin-8 and polymorphonuclear elastase levels in human septic shock.

Stress doses of hydrocortisone are known to have immunomodulatory effects in patients with hyperdynamic septic shock. The prognosis correlates with the presence and severity of septic encephalopathy. However, neurological evaluation is influenced by the use of analgesia sedation during artificial ventilation. The objective of this study was to demonstrate the effect of stress doses of hydrocortisone during the initial phase of human septic shock on the serum values of the neurospecific protein S-100B in comparison to the inflammation markers interleukin (IL)-8 in serum and polymorphonuclear (PMN) elastase in plasma. A total of 24 consecutive patients, who met the American College of Chest Physicians/Society of Critical Care Medicine criteria for septic shock, were enrolled in this prospective, randomized, double-blind, single-center trial. The severity of illness at recruitment was graded using the Acute Physiology and Chronic Health Evaluation II and the Simplified Acute Physiology Score II scoring systems. Multi-organ dysfunction syndrome was described by the Sepsis-related Organ Failure Assessment (SOFA) score. All patients were prospectively randomized to receive either stress doses of hydrocortisone or placebo. Hydrocortisone was started in 12 patients with a loading dose of 100 mg and followed by a continuous infusion of 0.18 mg/kg/h for 6 days. Median S-100B serum levels of the hydrocortisone group decreased from 0.32 ng/mL at study entry to 0.07 ng/mL 6 days later without significant differences compared to the placebo group. Initial IL-8 serum levels were significantly higher in the hydrocortisone group up to 12 h after study entry, and significantly decreased from 715 to 17 pg/mL at the end of the observation period. Median PMN elastase plasma levels were not affected by hydrocortisone infusion. Patients with initial S-100B serum levels > 0.50 ng/mL revealed significantly higher SOFA scores up to 30 h, IL-8 serum levels up to 12 h, and PMN elastase plasma levels up to 36 h after study entry than those patients with < or = 0.50 ng/mL. These effects were independent of the amount of fluid correction for hemodilution. Starting S-100B, IL-8 and PMN elastase values of the hydrocortisone group were within the ranges already known in patients with out-of-hospital cardiac arrest or severe traumatic brain injury. Stress doses of hydrocortisone resulted in a significant reduction in IL-8 serum, but not in S-100B serum and PMN elastase plasma concentrations in patients with hyperdynamic septic shock. For the first time, a similar extent of S-100B increase in serum of septic patients at the time of diagnosis was shown as reported for cardiac arrest or severe traumatic brain injury.

Adult↗

PubMed2004

Susceptibility of automated external defibrillators to train overhead lines and metro third rails.

INTRODUCTION: Immediate accessibility to automated external defibrillators (AED) is recommended for highly frequented public areas. In train terminals and metro stations electromagnetic interference (EMI) is present. In preparation for a public access defibrillation (PAD) programme in this environment possible effects on AED safety and accuracy were studied. METHODS: In typical public transportation settings 11 different AED models were bench tested for their sensitivity and specificity of ECG analysis with shockable and nonshockable rhythms provided by an ECG simulator. The devices were exposed to the electromagnetic interference of a rail system operating with 15 kV alternating current (ac) with a frequency of 16 2/3 Hz and a subway system powered with 750 V direct current (dc). AED cables were setup parallel and perpendicular to the tracks, the tests were carried out at 3 m distance from the rails in an empty station and with incoming trains. RESULTS: A total of 5280 tests were recorded, each device was tested a total of 480 times. Fifteen kilovolts 16 2/3 Hz ac interfered more than 750 V dc with the tachyarrhythmia detection systems (P < 0.0001). An AED setup with electrode cables perpendicular to track and power line reduced interference (P < 0.0001), while incoming trains had no significant effect on ECG analysis (P = 0.19). Depending on the AED model, sensitivity ranged from 60 to 100% and specificity from 54 to 100%, representing a positive likelihood-ratio from 1.3 to 241 and a negative likelihood-ratio from 0.7 to 0.0. In the public transportation setting tested, four AED models were unsuitable for automated defibrillation as these devices demonstrated an unacceptable performance in respect of accuracy and safety. In the train setting two devices performed with an accuracy of 57 and 65%. One AED recommended shocks for sinus rhythm at normal frequency. In the metro setting one AED did not advise shocks for ventricular tachycardia. CONCLUSION: Shock advisory systems of some AED models are susceptible to electromagnetic interference, especially in terminals with 15 kV 16 2/3 Hz ac power supplies. Interference is minimized, if patient position is parallel and electrode cables are perpendicular to overhead line. The choice of AED model for train or metro stations depends on its lack of susceptibility to typical electromagnetic interference.

Defibrillators↗

PubMed2004

Effect of digital cellular phones on tachyarrhythmia analysis of automated external defibrillators.

OBJECTIVES: Emergency services personnel, family members, laypersons or patients often carry and use mobile phones on sites of emergencies. As there are reported effects on implanted pacemakers and cardioverter defibrillators, the influence of digital cellular phones on automated external defibrillators was studied. METHODS: Twelve automated external defibrillator models were bench tested for their correct decision to or not to advise a shock, while being exposed to electromagnetic interference from a handheld cellular phone with 2 W or a portable cellular phone with 8 W transmitting power. The phones were programmed by a special subscriber identity module card to maximum output power with a carrier frequency of 906.2 MHz. The tests were conducted with a burst frequency of 217 Hz in speech mode and 2-8 Hz in discontinuous transmitting exchange mode. The sensitivity and specificity of electrocardiogram analysis systems were tested, with shockable and non-shockable rhythms provided by an electrocardiogram simulator and on two human subjects with normal sinus rhythm. RESULTS: A total of 8640 tests were recorded, each automated external defibrillator was tested a total of 720 times. The automated external defibrillators demonstrated a sensitivity of 100% and a specificity of 100%, representing a positive likelihood ratio of 8641 and a negative likelihood ratio of 0.000. In this setting all automated external defibrillators analysed correctly even under worst-case testing conditions, and performed excellently without any single failure. In some devices, voice prompts were distorted beyond comprehension, as the coil of the automated external defibrillator speaker received the pulsed signals. CONCLUSION: Shock advisory systems of automated external defibrillators are not susceptible to electromagnetic interference of 900 MHz cellular phones. Voice prompts, however, could be distorted by the operation of nearby digital mobile phones. During automated external defibrillator training this issue needs to be addressed.

Cell Phone↗

PubMed2004

5-Aminoisoquinolinone, a novel inhibitor of poly(adenosine disphosphate-ribose) polymerase, reduces microvascular liver injury but not mortality rate after hepatic ischemia-reperfusion.

OBJECTIVE: The aim of this study was to investigate the impact of the novel, potent, water-soluble inhibitor of poly(adenosine diphosphate-ribose) polymerase (PARP) 5-aminoisoquinolinone (5-AIQ) on hepatic microcirculation, hepatocellular injury, and survival in a murine model of hepatic ischemia-reperfusion. DESIGN: Randomized animal study. SETTING: Research laboratory. SUBJECTS: C57BL6 mice were subjected to warm either partial (90 mins) or total (75 mins) ischemia of the liver. INTERVENTIONS: Either PARP inhibitor 5-AIQ (3 mg/kg) or vehicle was administered to mice intravenously immediately before the start of reperfusion. Sham-operated animals served as controls. MEASUREMENTS AND MAIN RESULTS: As shown by intravital fluorescence microscopy after 30-60 mins of reperfusion, ischemia-reperfusion significantly enhanced platelet- and leukocyte-endothelial cell interactions in hepatic microvessels and impaired sinusoidal perfusion. Hepatocellular injury was characterized by an increase in the number of necrotic and apoptotic cells, dramatic elevation of aspartate aminotransferase/alanine aminotransferase serum activity, and lipid peroxidation in liver tissue. 5-AIQ treatment attenuated ischemia-reperfusion-induced increases in the numbers of adherent platelets and leukocytes as well as of necrotic and apoptotic cells and ameliorated perfusion failure. Furthermore, PARP inhibition prevented the increase in aspartate aminotransferase activity after ischemia-reperfusion but did not affect postischemic alanine aminotransferase release. However, no protective impact of 5-AIQ on postischemic oxidative stress was observed. Although PARP inhibition did not alter the survival percentage after ischemia-reperfusion (22% in both groups), this approach prolonged survival from 1 to 24 hrs (ischemia-reperfusion + vehicle) up to 48-72 hrs in the treated group. CONCLUSIONS: PARP inhibition with 5-AIQ during hepatic ischemia-reperfusion attenuates microvascular injury and reduces the extent of necrotic/apoptotic cell damage but does not protect from oxidative injury and does not improve postoperative survival rate.

Animals↗

PubMed2004

Initial posttraumatic translocation of NF-kappaB and TNF-alpha mRNA expression in peripheral blood monocytes of trauma patients with multiple injuries: a pilot study.

Post-traumatic inflammation is connected to monocyte dysfunction characterized by reduced NF-kappaB translocation during the first post-traumatic days. Because the exact dynamic of monocytic NF-kappaB translocation in patients directly after trauma remains unclear, the aim of this pilot study was to measure the intranuclear presence of NF-kappaB in monocytes from patients with multiple injuries initially after the trauma and during the early post-traumatic period and to compare these results with downstream-placed mRNA expression alteration of TNF-alpha, as well as with clinical data. Eleven patients were enrolled with an Injury Severity Score of 16 to 66 points, and blood samples were drawn on admission within 90 min and at 6, 12, 24, 48, and 72 h after trauma. NF-kappaB translocation of monocytic nuclear protein was analyzed by electrophoretic mobility shift assay and was quantified by densitometry as arbitrary units. In addition, monocytes of healthy volunteers were analyzed either native (-, control) or after LPS stimulation (+, control). For determination of downstream mRNA encoding for TNF-alpha, quantitative reverse transcriptase-PCR was performed. For both parameters, the negative control values were set as baseline (=1) and results from positive controls and patients were given as a relative alteration ratio without unit. Initial post-traumatic NF-kappaB translocation was significantly increased in trauma patients on admission (88 +/- 37) and 6 h after trauma (59 +/- 28) compared with the baseline level. In contrast, TNF-alpha mRNA was not increased on admission (1.7 +/- 0.9) and decreased even below baseline after 12 h. The substantial information of our study arises from the analysis of the dynamic of NF-kappaB translocation of monocytes. Enabled by closely matched sequential blood sampling strictly standardized to the traumatic event, an essential increase of monocytic signal transduction and transcription could be elucidated in the very early post-traumatic period, which precedes the down-regulation of the innate immune system.

Adolescent↗

PubMed2003

Initial RNA expression in human monocytes after multiple injury: a screening pilot study on potentially trauma-sensitive factors by using the microarray-technique.

BACKGROUND: Pathological affection of the immune system is one of the initiating mechanisms for the induction of multiple organ failure (MOF) in patients suffering from multiple injuries. Potential responsible intracellular mechanisms such as initial monocyte mRNA expression of specific mediators remain poorly studied, so far. Hence, we applied the microarray technique for screening of a wide variety of genes in circulating monocytes of multiple injured patients and compare the molecular results to the clinical course of the patients (MOF-score). METHODS: In our prospective pilot study 6 patients were enclosed presenting with blunt multiple injuries (Injury Severity Score 16 to 57 points). Monocytes were isolated out of sequentially drawn samples (6, 12, 24 and 48 hours after trauma) using magnetic cell sorting (CD14) and a human microarray system was used (Atlas stress 1.2, Clontech, 1176 genes). Alterations in the sequential samples were identified by calculating ratios to baseline levels on admission and cluster analysis was performed (Spotfire Decision). RESULTS: Only 86 (ca.5%) genes displayed an obvious signal. The house-keeping genes clustered well together in all patients in contrast to a substantial inter-individual variability of the other signal giving genes. No mediator burst of the classical pro- or anti-inflammatory cascade were detected. CONCLUSION: We demonstrate for the first time a screening analysis of mRNA expression patterns in circulating monocytes of multiple injured patients indicating that only very few genes appeared to be influenced by the traumatic event. So far, no correlation to the severity of trauma or MOF could be detected.

Adult↗

PubMed2003

Impact of intraischemic temperature on oxidative stress during hepatic reperfusion.

This study was designed to investigate the influence of intraischemic liver temperature on oxidative stress during postischemic normothermic reperfusion. In C57BL/6 mice, partial hepatic ischemia was induced for 90 min and intraischemic organ temperature adjusted to 4 degrees C, 15 degrees C, 26 degrees C, 32 degrees C, and 37 degrees C. As detected by electron spin-resonance spectroscopy, plasma/blood concentrations of hydroxyl and ascorbyl radicals were significantly increased in all groups after ischemia/reperfusion independent of the intraischemic temperature. In tissue, however, postischemic lipid peroxidation was attenuated after organ cooling down to 32 degrees C-26 degrees C and not detectable after ischemia at 15 degrees C-4 degrees C. mRNA expression of superoxide dismutase-1 and heme oxygenase-1, measured during reperfusion, was significantly elevated in the group at 37 degrees C as compared to the hypothermic groups at 4 degrees C-32 degrees C. The reduction of radical generation was associated with a prevention of adenosine monophosphate hydrolysis during ischemia in the hypothermic groups. In conclusion, ischemia-reperfusion-induced oxidative stress in the liver tissue is non-linearly-dependent on intraischemic temperature, whereas the plasma/blood concentration of radicals is not affected by organ cooling. Oxidative stress is reduced through mild hypothermia at 32 degrees C-26 degrees C and inhibited completely at 15 degrees C. Reduction of initial intracellular radical generation and prevention of secondary oxidant-induced tissue injury are possible mechanisms of this protection.

Adenosine Monophosphate↗

PubMed2003

Platelet-endothelial cell interactions during hepatic ischemia-reperfusion in vivo: a systematic analysis.

Platelets have been implicated in the pathophysiology of ischemia-reperfusion (I/R) injury. This study was designed to analyze platelet-endothelial cell interactions after I/R of the liver in vivo in dependence of the duration of ischemia and reperfusion. In C57BL/6 mice, a warm (37 degrees C) lobar hepatic ischemia was induced for 30, 60, or 90 min. Platelets were isolated from syngeneic donors, labeled ex vivo by rhodamine-6G, and infused intraarterially. Platelet-endothelial cell interactions and sinusoidal perfusion were analyzed using intravital fluorescence microscopy after 20-240 min of reperfusion. Hepatic I/R induced platelet rolling and adhesion in terminal arterioles and postsinusoidal venules as well as platelet accumulation in sinusoids already after 20 min of reperfusion. The number of platelets interacting with the endothelium was significantly increased as ischemia time was prolonged from 30 min up to 60 and 90 min. Postischemic platelet adhesion was associated with an increase in thrombin activity and a loss of platelets from the systemic circulation. Moreover, I/R-induced platelet adhesion was linearly correlated with an impairment of sinusoidal perfusion. The prolongation of reperfusion time up to 4 h did not further enhance platelet-endothelial cell interactions. These in vivo results indicate that hepatic I/R induces platelet-endothelial cell interactions in arterioles, sinusoids, and venules already during early reperfusion. The extent of these interactions is dependent on the duration of ischemia, but not on reperfusion time. Adherent platelets seem to participate in the development of sinusoidal perfusion failure in the postischemic liver.

Animals↗

PubMed2003

Microcirculation associated with degenerative rotator cuff lesions. In vivo assessment with orthogonal polarization spectral imaging during arthroscopy of the shoulder.

BACKGROUND: Diminished vascular supply is associated with degenerative rotator cuff lesions. Orthogonal polarization spectral imaging allows noninvasive assessment of microcirculation without application of fluorescent contrast medium. The aim of our study was to visualize and quantify in vivo the microcirculation of the rotator cuff during arthroscopic surgery and to compare the results with the number of microvessels identified in vitro by immunostaining of biopsy specimens taken from the scanned areas. METHODS: Eleven patients with clinical signs of a degenerative rotator cuff lesion were studied. Prior to arthroscopic subacromial decompression, the superficial part of the supraspinatus tendon at the edge of the lesion as well as the unaffected tendon insertion was examined. Microvascular parameters established for the description of tissue perfusion with use of conventional intravital fluorescence microscopy (functional capillary density and capillary diameter) were assessed in vivo. Biopsy specimens were taken from the scanned areas, and the microvessels were localized by immunostaining for the endothelial surface marker CD31. RESULTS: In the region of the unaffected tendon insertion, the mean baseline functional capillary density (and standard deviation) was 106 +/- 13 cm/cm(2) and the mean capillary diameter was 10 +/- 0.7 microm. In contrast, at the edge of the lesion, the functional capillary density was significantly reduced to 20 +/- 14 cm/cm(2), whereas the diameter of the vessels that were present did not differ. The total number of vessels stained in vitro was also significantly reduced at the edge of the lesion compared with the number of vessels in the tendon insertion zone. CONCLUSIONS: Quantitative in vivo analysis of human microcirculation during arthroscopy demonstrated that the functional capillary density at the edge of a degenerative rotator cuff lesion was significantly reduced compared with that in the control tissue. The capacity to assess microcirculatory flow in vivo may help to identify specific repair strategies based on knowledge of individual perfusion patterns.

Arthroscopy↗

PubMed2002

Platelet adhesion mediated by fibrinogen-intercelllular adhesion molecule-1 binding induces tissue injury in the postischemic liver in vivo.

BACKGROUND: Platelets are thought to be involved in the induction of hepatic ischemia-reperfusion (I/R) injury. The mechanisms of platelet adhesion in the hepatic microvasculature and the role of platelets in the pathogenesis of I/R-induced liver damage in vivo remain unclear. METHODS: In C57BL/6 mice, platelet- and leukocyte-endothelial cell interactions were quantitatively analyzed using intravital fluorescence microscopy in sham-operated animals, after warm lobar hepatic I/R (90/20 min) in wild-type and intercellular adhesion molecule (ICAM)-1-deficient mice, and after I/R in wild-type mice treated with an antifibrinogen antibody. Fibrinogen deposition on the endothelium was detected by intravital microscopy and by immunostaining. Reperfusion injury was assessed by measurement of liver enzyme and caspase-3 activities and of lipid peroxidation. RESULTS: Hepatic I/R induced fibrinogen deposition on hepatic endothelium, followed by a dramatic increase in the number of firmly adherent platelets in the liver microvasculature. Simultaneously, the number of adherent leukocytes in postsinusoidal venules and the aspartate aminotransferase/alanine aminotransferase and caspase-3 activities were elevated. Although ICAM-1 deficiency attenuated postischemic adherence of both platelets and leukocytes, the application of an antifibrinogen antibody selectively reduced the number of adherent platelets but did not influence leukocyte adhesion. The selective blockade of platelet adherence significantly prevented the postischemic increase in liver enzyme and caspase-3 activities. Furthermore, sinusoidal perfusion failure and lipid peroxidation were attenuated in the treated group. CONCLUSIONS: These in vivo data show that platelet adhesion mediated through fibrinogen deposition on ICAM-1 expressed on the endothelium of postischemic hepatic microvessels induces microvascular injury and hepatocellular apoptosis after I/R of the liver during early reperfusion.

Animals↗