Test-Retest Reliability of Kinematic Assessments for Upper Limb Robotic Rehabilitation.

Robot-measured kinematic variables are increasingly used in neurorehabilitation to characterize motor recovery following stroke. However, few studies have evaluated the reliability of these kinematic variables. This study aimed at evaluating the test-retest reliability of typically-used robot-measured kinematic variables in healthy subjects (HS) and patients with stroke (SP). Sixty-one participants (40 HS, 21 SP) carried out a planar robot-pointing task on two consecutive days. Nine robot-measured kinematic variables were computed: movement time (T), mean velocity (mV), maximal velocity (MV), smoothness error (SE), number of velocity peaks (nP), mean arrest period ratio (MAPR), normalized path length (NPL), root mean square error (RMS) from a straight line and the orthogonal projection of the last point of movement (LP). Intraclass Correlation Coefficients (ICC), percentage of the Standard Error of Measurement (SEM measured as a percentage of the mean value of the variable (%SEM)) and percentage of the Minimum Detectable Difference (MDD measured as a percentage of the mean value of the variable (%MDD)) were used to analyze the test-retest reliability of the kinematic variables. ICC scores for all kinematic variables were above 0.75 in both groups. %SEM values were below 10% except for MAPR (13.4%) in HS and nP, MAPR and RMS in SP (13.0%, 11.7% and 15.2% respectively). %MDD values were higher for RMS in SP (42.1%) and MAPR in HS (37.1%) and lower for LP (1.6% in HS and 8.1% in SP). The nine robot-measured kinematic variables all demonstrated good reliability, with high ICC values (>0.75) and an acceptable level of measurement error (%SEM< 16%). However, 3/9 robot-measured kinematic variables, did not appear to be sufficiently sensitive to change (%MDD>30%) to be considered useful in patients with stroke.

[Relevance of Vascular Trauma in Trauma Care - Impact on Clinical Course and Mortality]. / Bedeutung des Gefäßtraumas für die Schwerverletztenversorgung ­ Einfluss auf Verlauf und Mortalität.

There is a lack of evidence as to the relevance of vascular trauma (VT) in patients with severe injuries. Therefore, we reviewed registry data in the present study in order to systematically objectify the effect of VT in these patients. This study aimed to provide an adequate picture of the relevance of vascular trauma and to identify adverse prognostic factors. In a retrospective analysis of records from the TraumaRegister DGU® (TR-DGU) in two subgroups with moderate and severe VT, we examined the records for differences in terms of morbidity, mortality, follow-up and prognostic parameters compared to patients without VT with the same ISS. From a total of 42,326 patients, 2,961 (7 %) had a VT, and in 2,437 cases a severe VT (AIS ≥ 3) was diagnosed (5.8 %). In addition to a higher incidence of shock and a 2 to 3-fold increase in fluid replacement and erythrocyte transfusion, patients with severe VT had a 60 % higher rate of multiple organ failure, and in-hospital mortality was twice as high (33.8 %). The massively increased early mortality (8.0 vs. 25.2 %) clearly illustrates how severely injured patients are placed at risk by the presence of a relevant VT with a comparable ISS. In our opinion, due to an unexpected poor prognosis in the TR-DGU data for vascular injuries, increased attention is required in the care of severely injured patients. Based on our comprehensive analysis of negative prognostic factors, a further adjustment to the standards of vascular medicine could be advisable. The influence of the level of care provided by the admitting hospital and the relevance of a further hospital transfer to prognosis and clinical outcome is currently being analysed.

[Computed tomography angiography as the basis for optimized therapy planning before endovascular aneurysm repair (EVAR)]. / CT-Angiographie als Grundlage der optimierten Therapieplanung vor endovaskulärer Aneurysmaausschaltung (EVAR).

Computed tomography angiography (CTA) of the aorta is an accepted standard diagnostic procedure for preoperative evaluation and planning of endovascular treatment of abdominal aortic aneurysms (endovascular aortic repair EVAR). The CTA method delivers all relevant anatomical and morphological information on the underlying pathology of the aorta and pelvic axes. Various software solutions are available for multiplanar reconstruction of the CT data for exact measurement of the access routes and landing zones and are essential components of individualized operation planning. The synthesis of all CT-based information allows a safe and exactly targeted release of the stent graft in the aorta. Furthermore, the periprocedural radiation dose can be reduced by a precise preoperative planning of the positions to be irradiated during implantation.

[Emergencies in vascular surgery--a major challenge for the local infrastructure]. / Gefäßchirurgische Notfälle--eine große Herausforderung an die lokale Infrastruktur.

INTRODUCTION: Emergencies in vascular surgery are often life-threatening and require a timely and prompt treatment. Little information is available in the literature about which demands must be made for this on the personnel and infrastructural resources of a hospital. METHODS: All vascular surgical emergency operations of the Surgical University Hospital of Munich - Grosshadern over a period of 2 years were evaluated concerning the emergency category, the leading clinical symptomatology, the genesis, the affected stream area, the intervention time, as well as the need for postoperative intensive medical care. RESULTS: The prevailing procedures were arterial operations (76 %). Ischaemia with 37 % and bleeding with 29 % were the leading clinical symptomatology. Thrombotic events (34 %) showed the most frequent genesis followed by embolism (13 %), stenosis (11 %), aneurysms (10 %) and iatrogenic impairments (10 %). 68 % of the emergencies were treated outside of the daytime working hours. A total of 77 % of the patients needed intensive care treatment or observation after surgery. CONCLUSION: The spectrum and the frequency of emergencies in vascular surgery make high demands on local infrastructure of the hospital and require a fair number of intensive care beds and an adequate and highly trained staff. Only under these conditions can a high quality of treatment be guaranteed for the sometimes life-threatened patients.

microRNA expression signatures and parallels between monocyte subsets and atherosclerotic plaque in humans.

Small non-coding microRNAs (miRNAs) have emerged to play critical roles in cardiovascular biology. Monocytes critically drive atherosclerotic lesion formation, and can be subdivided into a classical and non-classical subset. Here we scrutinised the miRNA signature of human classical and non-classical monocytes, and compared miRNA expression profiles of atherosclerotic plaques from human carotid arteries and healthy arteries. We identified miRNAs to be differentially regulated with a two-fold or higher difference between classical and non-classical monocyte subsets. Moreover, comparing miRNA expression in atherosclerotic plaques compared to healthy arteries, we observed several miRNAs to be aberrantly expressed, with the majority of miRNAs displaying a two-fold or higher increase in plaques and only few miRNAs being decreased. To elucidate similarities in miRNA signatures between monocyte subsets and atherosclerotic plaque, expression of miRNAs highly abundant in monocytes and plaque tissues were compared. Several miRNAs were found in atherosclerotic plaques but not in healthy vessels or either monocyte subset. However, we could identify miRNAs co-expressed in plaque tissue and classical monocytes (miR-99b, miR-152), or non-classical monocytes (miR-422a), or in both monocytes subsets. We thus unravelled candidate miRNAs, which may facilitate our understanding of monocyte recruitment and fate during atherosclerosis, and may serve as therapeutic targets for treating inflammatory vascular diseases.

[Intraoperative neuromonitoring for prevention of neurological complications in aortic surgery]. / Intraoperatives Neuromonitoring zur Prävention neurologischer Komplikationen bei Aortenbogen- und thorakoabdomineller Aortenchirurgie.

AIM: Stroke and paraplegia are devastating complications of thoracic and thoracoabdominal aortic surgery. The aim of this study was to analyse the value of transcranial Doppler ultrasound (TCD), electroencephalogram (EEG) and motor-evoked potentials (MEP) in preventing neurological complications. Moreover, the principles, technology and surgical protocols are described. PATIENTS AND METHODS: In 2009, 22 patients (4 females, 18 males) underwent thoracic or thoracoabdominal open aortic repair. We performed 2 arches with descending aortic replacement, 5 arches with TAAA repair, 2 type II, 9 type III, 3 type IV and one type V TAAA aortic repair. In 6 patients, the neuromonitoring included TCD, EEG and MEPs. In 15 patients only MEP monitoring was necessary. In one patient who was operated on in an emergency setting, neuromonitoring was not performed. The surgical approach was a left thoracotomy in 3 and a left thoracolaparotomy in 19 patients. The surgical protocol included cerebrospinal fluid drainage (n=22), moderate (n=19) or deep hypothermia (n=2), and extracorporeal circulation (n=21) with retrograde aortic perfusion and selective cerebral and/or viscerorenal perfusion. RESULTS: In 21 patients, the neuromonitoring could be established successfully. Using TCD and EEG, a relevant cerebral ischaemia during supraaortic clamping was excluded. With a mean distal arterial pressure of 60 mmHg, the MEPs remained adequate in 15 patients (68.2%). Increasing of the blood pressure restored the MEPs in one patient. In 5 patients (22.7%), a reimplantation of segmental arteries (n=4) or of the left subclavian artery (n=1) re-established spinal cord perfusion, as indicated by restored MEPs. We had no absent MEPs at the end of the procedures. Delayed paraparesis developed in 2 patients with a haemodynamic instability during the postoperative course. Paraplegia was not observed. CONCLUSION: TCD, EEG and MEPs are reliable techniques to unmask cerebral or spinal cord ischaemia during aortic surgery. Immediate operative strategies based on neuromonitoring information prevent neurological complications in aortic surgery.

Early control of distal internal carotid artery during carotid endarterectomy: does it reduce cerebral microemboli?

AIM: According to the results of the large trials on carotid endarterectomy (CEA), this type of surgery is only warranted if perioperative mortality and morbidity are kept considerably low. Less attention has been paid to methods of cerebral protection during CEA, although intraoperative transcranial Doppler (TCD) can visualise intracerebral microemboli (MES) during routine carotid dissection, although MES occur throughout the CEA, only those during dissection are related to neurological outcome. Prevention of MES by means of early control of the distal internal carotid artery dislodging from the carotid artery plaque during dissection is very likely the mechanism behind an eventual benefit from this approach. Hence, the amount of MES might serve as a surrogate parameter for the risk of periprocedural neurological events. So, the aim of the present study was to evaluate whether early control of the distal carotid artery during CEA is capable of reducing the number of MES by means of a prospective randomised trial. METHODS: Twenty-eight patients (29 procedures) could be prospectively included in our study. Before surgery we randomly assigned the patients to two groups: group A (N.=12): CEA by means of early control of the distal internal carotid artery; group B (N.=17): CEA with dissection of the total carotid bifurcation before clamping the arteries. Periprocedurally, we continuously monitored the cerebral blood flow in the ipsilateral middle cerebral artery by means of TCD. Pre- and postoperative morbidity were independently verified by a neurologist <2 days before and not later than five days after the procedure. Values of microembolic signs during dissection were summarised with arithmetic means and standard deviations. For further analysis non parametric Wilcoxon test was performed between both methods. P-values <0.05 were considered as statistically significant. Wilcoxon test was performed to compare both methods concerning clamp- and procedure times. RESULTS: We performed EEA 26 times, in three patients a longitudinal arteriotomy with endarterectomy and patchplasty was performed, in one of these patients a shunt was necessary. In 12 twelve patients MES occurred during the dissection before clamping. Eight of these patients belonged to group B and four patients to group A. The mean number of MES during dissection for group A was 2.4 (SD 4.6; 5-15) and for group B 3.9 (SD 7.1; 2-28). There is no statistically significant difference in the Wilcoxon-test; P=0.4375. There was no patient showing reperfusion syndrom or clinical signs of a new cerebral infarction or any other neurological deficit. There were no other major complications like myocardial infarction or death as well as no minor complications like periphereal nerve lesions, bleeding or wound healing disturbance. CONCLUSION: In this prospective, randomised trial early control of the distal internal carotid artery did not reduce the occurrence of MES during dissection of the carotid bifurcation. Also, the total number of MES throughout the procedure and postoperatively was comparable between both groups. The procedure related times as well as the clinical outcome did not differ significantly. Thus, early control of the distal internal carotid artery has got no advantage but also no disadvantage as compared to the traditional CEA technique. However, a limitation of the study is the small number of patients included.

Motor evoked potentials in thoracoabdominal aortic surgery: PRO.

Paraplegia is one of the most severe complications of the repair of open descending thoracic aortic aneurysms and thoracoabdominal aortic aneurysms. To reduce these complications, a comprehensive strategy for spinal cord protection is mandatory. Motor evoked potentials provide the surgeon with important information about spinal cord integrity throughout the operation. Neuroprotective measures include extracorporeal circulation, cerebrospinal fluid drainage, hypothermia, and selective segmental artery revascularization.

Carotid plaque analysis: comparison of dual-source computed tomography (CT) findings and histopathological correlation.

PURPOSE: Plaque morphology is an important predictor of stroke risk and may also be a predictor of postoperative outcome after carotid endarterectomy (CEA). Thus, the purpose of our study was to evaluate the findings of preoperative dual-source computed tomography (DSCT) of carotid plaque morphology and correlate these findings with histopathological findings. MATERIAL AND METHODS: Thirty patients undergoing CEA due to neurological events and high-grade carotid artery stenosis were evaluated with DSCT for degree of stenosis following the North American Symptomatic Carotid Endarterectomy Trial (NASCET) criteria and for non-invasive plaque morphology prior to CEA. CT protocol was as follows (SOMATOM Definition, Siemens Medical Solutions, Forchheim, Germany): A dual-energy protocol was used with tube A (140 kV, 55 mA) and tube B (80 kV, 230 mA) with 2 x 64 x 0.6-mm collimation, pitch 0.65 and rotation time of 0.33 s. Histopathological work-up was performed on the surgically retrieved tissues. The findings from DSCT and histopathology were compared with respect to image quality and plaque composition (fatty plaque, mixed plaque and calcified plaque), were correlated with histological specimens and classified according to the American Heart Association (AHA) classification of atherosclerotic plaque. Pearson correlation and kappa statistics were performed. RESULTS: The image quality of DSCT was rated as 'excellent' in all the examinations. The mean degree of stenosis was quantified as 82%. The sensitivity of DSCT for the detection of calcification was 100% (standard deviation (SD) 0%, confidence interval (CI): 99-100). While the sensitivity for the detection of mixed plaques was 89% (SD 12%, CI: 79-98), it was 85% (SD 10%, CI: 76-92) for the detection of low-density fatty plaques. The mean degree of agreement was k=0.81. CONCLUSION: DSCT angiography of the carotid arteries is feasible and the evaluation of carotid plaque composition allows non-invasive assessment of different plaque components. This may have an impact on the non-invasive differentiation of vulnerable plaques.

The role of procalcitonin as predictor for neurological deficits after carotid endarterectomy.

AIM: Outcome of carotid endarterectomy (CEA) is defined by mortality rate as well as the neurological outcome due to cerebral ischemia. Thus the aim of this study was to evaluate the role of the acute phase protein procalcitonin (PCT) as a predictor for neurological deficits after carotid endarterectomy. METHODS: Fifty-five patients with high grade stenosis of the internal carotid artery and interdisciplinary consensus for endarterectomy were followed. Neurological examination was performed before and after the procedure to analyze perioperative neurological deficits. Blood samples were obtained before and after CEA and procalcitonin was analyzed in 55 consecutive patients (65.5% symptomatic/34.5% asymptomatic). RESULTS: No perioperative or in-hospital death was observed. Major complications did not occur, two patients suffered from bleeding requiring surgical intervention and one patient had a temporary peripheral facial nerve lesion. Postoperative neurological examination revealed no new deficit, there was no significant change of PCT (level pre- and post-CEA (the mean preoperative PCT was 0.25 ng/mL [SD 0.78, min 0.1, max 4.3]; the mean postoperative PCT was 0.11 ng/mL [SD 0.06, min 0.1, max 0.5]). There was no association found between perioperative neurological deficit and PCT. CONCLUSIONS: The present study demonstrates that there is still not sufficient evidence to recommend PCT measurement as a predictor for perioperative neurological deficit during CEA.

Thoracoabdominal aortic aneurysm repair in patients with marfan syndrome.

OBJECTIVE: We assessed the surgical outcome of descending thoracic aortic aneurysm repair (DTAA) and thoracoabdominal aortic aneurym (TAAA) repair in patients with Marfan syndrome. METHODS: During a six year period, 206 patients underwent DTAA and TAAA repair. In 22 patients, Marfan syndrome was confirmed. The median age was 40 years with a range between 18 and 57 years. The extend of the aneurysms included 6 DTAA (1 with total arch, 2 with distal hemi-arch), 11 type II TAAA (2 with total arch, 3 with distal hemi-arch), 4 type III and one type IV TAAA. All patients suffered from previous type A (n=6) or type B (n=16) aortic dissection and 15 already underwent aortic procedures like Bentall (n=7) and ascending aortic replacement (n=8). All patients were operated on according to the standard protocol with cerebrospinal fluid drainage, distal aortic and selective organ perfusion and monitoring motor evoked potentials. In patients undergoing simultaneous arch replacement (via left thoracotomy), transcranial Doppler and EEG assessed cerebral physiology during antegrade brain perfusion. In four patients circulatory arrest under moderate hypothermia was required. RESULTS: In-hospital mortality did not occur. Major postoperative complications like paraplegia, renal failure, stroke and myocardial infarction were not encountered. Mean pre-operative creatinine level was 125mmol/L, which peaked to a mean maximal level of 130 and returned to 92mmol/L at discharge. Median intubation time was 1.5 days (range 0.33-30 days). Other complications included bleeding requiring surgical intervention (n=1), arrhythmia (n=2), pneumonia (n=2) and respiratory distress syndrome (n=1). At a median follow-up of 38 months all patients were alive. Using CT surveillance, new or false aneurysms were not detected, except in one patient who developed a visceral patch aneurysm six years after open type II repair. CONCLUSION: Surgical repair of descending and thoracoabdominal aortic aneurysms provides excellent short- and mid-term results in patients with Marfan syndrome. In this series, a surgical protocol with cerebrospinal fluid drainage, distal aortic and selective organ perfusion and monitoring motor evoked potentials resulted in low morbidity and absent mortality. These outcomes of open surgery should be considered when discussing endovascular aneurysm repair in Marfan patients.

Differences of venous thromboembolic risks in vascular general and trauma surgery patients.

AIM: Venous thromboembolism (VTE) is a common complication in patients undergoing surgery. The risk for VTE is determined by the combination of individual predisposing factors and features of the specific type of surgery. Although the knowledge about VTE has increased enormously during the last years VTE-prophylaxis is still inadequate. The goals of our study were to assess the correctness of the adjusted pharmacological prophylaxis, and the difference of the VTE-risks in the different surgical departments. METHODS: During a three months period, 451 patients were prospective included. These patients were admitted to the Departments of Vascular and General Surgery and of Traumatology of our hospital. Based on the modified Hertfelder's VTE-risk-assessment model, we scored the patients and categorized them into 4 groups: low, moderate, high and very high risk for VTE. We enrolled every admitted patient taking their medical history and reviewing medical documents. RESULTS: The mean cumulative risk value for VTE-risk was 3.68 (median 3.5, minimum: 0, maximum: 13 and standard deviation: 2.206), whereas 20.2% of our patients had a low, 27.2% middle, 21.7% high and 30.9% very high risk. The patients with vascular procedures had significantly higher mean value (5.03, SD 2.2) than the patients with general operations (3.6, SD 2.2) and those who underwent traumatology (3.06, SD 1,8) (P value <0.001). The majority of patients (n=356), (78.9%) received VTE-prophylaxis with low dose of low molecular weight heparin (LMWH). Of the remaining patients, 40 (8.9%) received therapeutic dose and 55 (12.2%) received none VTE-prophylaxis. CONCLUSION: The VTE-risk for surgical patients remains high, despite all efforts for prophylaxis. The main reason may be that risk-assessment is time consuming and not standardized. We demonstrated that VTE-risk for patients in vascular surgery is significantly higher than the VTE-risk for patients in general and trauma surgery. We also showed that the VTE-risk in some patients was underestimated and prophylaxis was inadequate. Therefore, it is recommended to emphasize more on short risk-assessment, adequate prophylaxis and optimal dosage in order to prevent deep venous thrombosis and embolism disease.

Surgical repair of thoracoabdominal aortic aneurysms.

Morbidity and mortality following thoracoabdominal aortic aneurysm (TAAA) repair are tremendous. Preoperative assessment is essential in detecting cardiac and pulmonary risk factors in order to reduce cardiopulmonary complications. Paraplegia and renal failure are main determinants of postoperative mortality and therefore gained substantial attention during the last decades. Left heart bypass, cerebrospinal fluid (CSF) drainage and epidural cooling have significantly reduced paraplegia rate, however, this dreadful event still occurs in up to 25% of patients undergoing type II repair. Renal failure has been partly prevented by means of retrograde aortic perfusion and cooling but renal failure still remains a significant problem. We have evaluated the effects of protective measures aiming for reduction of paraplegia and renal failure. Monitoring motor evoked potentials (MEPs) is an accurate technique to assess spinal cord integrity during TAAA repair, guiding surgical strategies to prevent paraplegia. Selective volume- and pressure controlled perfusion is a technique to continuously perfuse the kidneys during aortic cross clamping and subsequent circulatory exclusion In patients with atherosclerotic thoracoabdominal aortic aneurysms, blood supply to the spinal cord depends on a highly variable collateral system. In our experience, monitoring MEPs allowed detection of cord ischemia, guiding aggressive surgical strategies to restore spinal cord blood supply and reduce neurologic deficit: overall paraplegia rate was less than 3%. We believe that these protective measures should be included in the surgical protocol of TAAA repair, especially in type II cases. Renal and visceral ischemia can be reduced significantly by continuous perfusion during aortic cross clamping in TAAA repair. Not only sufficient volume flow but also adequate arterial pressure appears to be essential in maintaining renal function.Obviously, endovascular modalities have been successfully applied in TAAA patients, the majority of which as part of hybrid procedures. Technological innovation will eventually cause a shift from open to minimal invasive surgical repair. At present, however, open surgery is considered the gold standard for TAAA repair, especially in (relatively) young patients and patients suffering from Marfan's disease.

[Carotid-surgery in ultrasound-guided anesthesia of the regio colli lateralis]. / Ultraschallgestützte Blockade der Regio colli lateralis zur Karotis-Chirurgie -- eine Fallserie.

OBJECTIVE: Carotid endarterectomy significantly reduces the risk of stroke in patients with symptomatic and asymptomatic carotid artery stenosis. An increasing number of interventions in carotid surgery are performed under regional anesthesia in conscious patients. Carotid endarterectomy in local anesthesia requires block of the cervical nerves C2-C4, which may be accomplished in different ways. The most frequent method of regional anesthesia in carotid surgery is a combined block of profunda and superficial cervical plexus (by using a nerve stimulator). Ultrasound is frequently used in anesthesia for venous access and peripheral nerve block. By ultrasound, it is possible to visualize puncture needle and spread of local anesthetics at the correct position for block of cervical plexus in carotid surgery. The aim of the case reports was to demonstrate the effectiveness of using ultrasound for regional anesthesia in carotid surgery. METHOD: At the level of carotid bifurcation, scalene muscles were visualized by ultrasound (10-MHz-transducer). At this position, a puncture needle was inserted in the beam of ultrasound. Between the anterior scalene and the sternocleidomastoid muscles the local anesthetics were injected (10 ml prilocaine 1 %, 20 ropivacaine 0.375 %). In addition, superficial cervical plexus was performed by subcutaneous injection (10 ml prilocaine 1 %, 40 ropivacaine 0.375 %). Sedation was performed by application of remifentanil (max. 0.02 microg/kg/min), as needed. RESULTS: Ultrasound guided cervical block was performed in 29 patients. In all cases cervical plexus block was complete. A conversion to general anesthesia during operation was not necessary in any case. CONCLUSION: Ultrasound presents an alternative to cervical block using nerve stimulation, especially in patients with anatomical abnormalities. The method proves to be effective and may improve the approach to profundeal cervical plexus.

Impact of inhibition of complement by sCR1 on hepatic microcirculation after warm ischemia.

Recent observations provide evidence that complement is implicated as an important factor in the pathophysiology of ischemia/reperfusion injury (IRI). Here, we assessed the effects of complement inhibition on hepatic microcirculation by in vivo microscopy (IVM) using a rat model of warm hepatic ischemia clamping the left pedicle for 70 min. Ten animals received the physiological complement regulator soluble complement receptor type 1 (sCR1) intravenously 1 min prior to reperfusion. Controls were given an equal amount of Ringer's solution (n = 10). Microvascular perfusion and leukocyte adhesion were studied 30 to 100 min after reperfusion by IVM. Microvascular perfusion in hepatic sinusoids was significantly improved in the sCR1 group (80.6 +/- 0.6% of all observed sinusoids were perfused [sCR1] vs 67.3 +/- 1.2% [controls]). The number of adherent leukocytes was reduced in sinusoids (49.9 +/- 3.4 [sCR1] vs 312.3 +/- 14.2 in controls [adherent leukocytes per square millimeter of liver surface]; P < 0.001) as well as in postsinusoidal venules after sCR1 treatment (230.9 +/- 21.7 [sCR1] vs 1906.5 +/- 93.5 [controls] [adherent leukocytes per square millimeter of endothelial surface]; P < 0.001). Reflecting reduced hepatocyte injury, liver transaminases were decreased significantly upon sCR1 treatment compared to controls. Our results provide further evidence that complement plays a decisive role in warm hepatic IRI. Therefore, we conclude that complement inhibition by sCR1 is effective as a therapeutical approach to reduce microcirculatory disorders after reperfusion following warm organ ischemia.

Complement inhibition by soluble complement receptor type 1 improves microcirculation after rat liver transplantation.

BACKGROUND: Recent observations provide evidence that complement is involved in the pathophysiology of ischemia/reperfusion injury. In this study, we assessed the impact of complement inhibition on hepatic microcirculation and graft function using a rat model of liver transplantation. METHODS: Arterialized orthotopic liver transplantation was performed in Lewis rats after cold preservation (University of Wisconsin solution, 4 degrees C, 24 h). Eight animals received the physiological complement regulator soluble complement receptor type 1 (sCR1) intravenously 1 min before reperfusion. Controls received Ringer's solution (n=8). Microvascular perfusion, leukocyte adhesion, and Kupffer cell phagocytic activity were studied 30-100 min after reperfusion by in vivo microscopy. RESULTS: Microvascular perfusion in hepatic sinusoids was improved in the sCR1 group (87+/-0.7% vs. 50+/-1%; P < 0.001). The number of adherent leukocytes was reduced in sinusoids (68.3+/-4.7 vs. 334.1+/-15.8 [adherent leukocytes per mm < or = liver surface]; P < 0.001) and in postsinusoidal venules after sCR1 treatment (306.6+/-21.8 vs. 931.6+/-55.9 [adherent leukocytes per mm < or = endothelial surface]; P < 0.001). Kupffer cell phagocytic activity was decreased in the sCR1 group compared to controls. Postischemic bile production reflecting hepatocellular function was increased by almost 200% (P = 0.004) after complement inhibition. Plasmatic liver enzyme activity was decreased significantly upon sCR1 treatment, indicating reduced parenchymal cell injury. CONCLUSIONS: Our results provide further evidence that the complement system plays a decisive role in hepatic ischemia/reperfusion injury. We conclude that complement inhibition by sCR1 represents an effective treatment to prevent reperfusion injury in liver transplantation.

Role of glutathione in hepatic bile formation during reperfusion after cold ischemia of the rat liver.

BACKGROUND/AIMS: Liver reperfusion following cold ischemia is frequently associated with diminished bile flow in patients undergoing liver transplantation. Glutathione is a major determinant of bile-acid independent bile flow, and the effects of cold ischemia on biliary glutathione excretion are unknown. METHODS: We examined the effects of cold ischemia (University of Wisconsin solution (4 degrees C), 24 h) with subsequent reperfusion (100 min) on biliary glutathione excretion in a recirculating system. Since glutathione might represent an important antioxidant within the biliary tract and oxidative stress in the biliary tract during reperfusion could contribute to the pathogenesis of bile duct injury after liver transplantation, we also assessed bile duct morphology in reperfused livers of mutant TR- -rats, in whom biliary excretion of glutathione is already impaired. RESULTS: Hepatic bile formation was diminished in reperfused Wistar rat livers after cold ischemia. Biliary glutathione concentrations and output were significantly decreased and correlated with postischemic changes in bile secretion. An increased biliary oxidized glutathione/glutathione ratio, indicating oxidative stress, was detected only immediately after the onset of reperfusion. Basal bile flow rates in TR- -rat livers which were already markedly reduced in control-perfused livers, decreased further during the early but not the later reperfusion period. Reperfusion of both Wistar and TR- -rat livers was not associated with electron microscopic evidence of bile duct damage. CONCLUSIONS: We conclude that impaired biliary excretion of glutathione contributes to decreased bile flow after cold ischemia. The absence of biliary glutathione does not appear to promote ultrastructural evidence of bile duct injury during reperfusion in the isolated perfused rat liver.

Extrahepatic biliary obstruction impairs microvascular perfusion and increases leukocyte adhesion in rat liver.

To determine if disturbances of the liver microcirculation may be of pathophysiological relevance for liver damage during acute biliary obstruction, we studied the effects of bile duct ligation (BDL) on hepatic microhemodynamics and leukocyte adhesion in rat liver in vivo. Male Wistar rats were subjected to BDL for 3 days and 7 days, respectively. Sham-operated controls underwent laparotomy without BDL. After 3 days, intravital fluorescence microscopy (IVM) and hydrogen gas (H2) clearance were performed to study hepatic microvascular perfusion. Furthermore, leukocyte-endothelial cell interactions were assessed by IVM. Intercellular adhesion molecule 1 (ICAM-1) protein expression was studied by Western blot analysis and tissue immunofluorescence after 3 and 7 days, respectively. Analysis of microvascular perfusion by IVM revealed a marked impairment of sinusoidal perfusion after 3 days. Assessment of H2 clearance confirmed that overall hepatic microvascular perfusion was decreased. In addition, increased leukocyte adhesion in sinusoids and venules could be observed. A concomitant increase of ICAM-1 expression in liver tissue was also noted within the first week after BDL. Our results show that BDL is followed by a marked depression of the hepatic microcirculation and increased leukocyte adhesion in vivo within 3 to 7 days. Together, these findings suggest that deficits in microvascular perfusion and increased neutrophil infiltration may represent a potential source of liver injury during acute biliary obstruction.

The rat canalicular conjugate export pump (Mrp2) is down-regulated in intrahepatic and obstructive cholestasis.

BACKGROUND & AIMS: The excretion of various organic anions into bile is mediated by an adenosine triphosphate-dependent conjugate export pump, which has been identified as the canalicular isoform of the multidrug resistance protein (Mrp2). Mrp2 function is impaired in various experimental models of intrahepatic and obstructive cholestasis, but the underlying molecular mechanisms are unclear. The aim of this study was to investigate these molecular mechanisms. METHODS: The effects of endotoxin, ethinylestradiol, and common bile duct ligation (CBDL) on Mrp2 protein, messenger RNA (mRNA) expression, and Mrp2 tissue localization were determined in rat livers by Northern blotting, Western analysis, and tissue immunofluorescence. To assess whether changes were specific for Mrp2, we also examined the expression of canalicular ecto-adenosine triphosphatase (ecto-ATPase) and mdr P-glycoproteins (P-gp). RESULTS: All three cholestatic models resulted in a marked decrease in Mrp2 protein (P < 0.01) and its tissue localization at the canalicular membrane. Mrp2 mRNA levels diminished profoundly after endotoxin (P < 0.0005) and CBDL (P < 0.05), but did not change after ethinylestradiol. In contrast to Mrp2, protein expression of ecto-ATPase and P-gp remained unchanged in endotoxin- and ethinylestradiol-treated animals, whereas P-gp levels increased after CBDL (P < 0.05). CONCLUSIONS: Down-regulation of Mrp2 expression may explain impaired biliary excretion of amphiphilic anionic conjugates in these models of cholestasis.