Prospective randomized trial to determine the influence of laparoscopic and conventional colorectal resection on intravasal fibrinolytic capacity

Background: Although the pneumoperitoneum decreases venous reflux from the lower extremities, the rate of thromboembolic complcations seems to be lower after laparoscopic than after conventional procedures. Therefore, it has been assumed that laparoscopic surgery better preserves the intravasal fibrinolytic capacity. The aim of this study was to determine the influence of the operative technique on intravasal fibrinolytic capacity in colorectal resection. Methods: Randomized controlled trial conducted in parallel with the multicenter trial LAPKON II comparing the long-term effects of elective laparoscopic (group I) and conventional (group II) resections for colorectal cancer. Blood samples were taken from 30 patients preoperatively, at the beginning and end of surgery as well as 2, 8, and 24 hr postoperatively. Activities and concentrations of tissue plasminogen activator (tPA), plasminogen activator inhibitor type 1 (PAI-1), tPA/PAI complex, fibrinogen, and D-dimers were determined in all specimen with ELISA tests. Area under the curve values (AUC) were calculated for all parameters. Results: Patient characteristics and indication for surgery were not different between both groups. Preoperative values of fibrinolytic parameters were similar in both groups. Postoperatively, tPA activity decreased significantly in both groups, but AUC values for tPA and PAI-1 activity (p = 0.23; p = 0.68); concentration of tPA, PAI-1, and tPA/PAI complex (p = 0.52; p = 0.78; p = 0.95); and concentration of fibrinogen and D-dimers (p = 0.67; p = 0.71) did not differ between the groups. Conclusions: An intravasal fibrinolytic "shutdown" occurs not only after conventional but also after laparoscopic colorectal resection. Both operative techniques had similar effects on postoperative intravasal fibrinolytic capacity. Therefore, the lower incidence of thromboembolic complications after laparoscopic colorectal resections does not seem to be caused by a lesser depression of the intravasal fibrinolytic capacity.     

Fibrinolytic responses of human peritoneal fluid in laparoscopic cholecystectomy: a prospective clinical study

Background The reduction in peritoneal fibrinolysis is believed to be the pathogenetic mechanism of adhesion formation. The general conclusion based on previous clinical and experimental studies is that laparoscopic procedures produce less adhesion formation. The association between this beneficial effect of laparoscopic cholecystectomy and peritoneal fibrinolytic changes is not clear. Therefore, the authors aimed to compare the effects of open and laparoscopic cholecystectomy on peritoneal fibrinolysis. For this purpose, fibrinolytic parameters in peritoneal fluid were investigated 24 h after laparoscopic and open cholecystectomies. Methods In a prospective clinical study, peritoneal fluid was sampled via a drain 24 h after laparoscopic (n = 10) and open (n = 9) cholecystectomies. Activities and concentrations of tissue plasminogen activator (tPA), plasminogen activator inhibitor type 1 (PAI-1), and tPA/PAI-1 complex were determined by enzyme-linked immunosorbent assay (ELISA) kits. Results In peritoneal fluids, tPA and tPA/PAI-1 complex concentrations were higher in the open cholecystectomy group (p = 0.009 and p < 0.001, respectively), but tPA activity and PAI-1 concentrations did not differ between the groups (p = 0.514 and p = 0.716, respectively). Conclusions Fibrinolytic changes in peritoneal fluid have several similarities in open and laparoscopic cholecystectomies with regard to tPA activity and PAI-1 levels. However, higher tPA levels after the open procedure probably are secondary to more intense tissue handling leading to mesothelial release of tPA.     

Hyaluronan levels during laparoscopic versus open colonic resections

Background Plasma hyaluronan binds to fibrinogen, affecting intravascular fibrin polymerization and fibrin clot formation. It has been hypothesized that alterations in fibrin clot formation influence the risk of thromboembolism in those undergoing surgery. The aim of this study is to quantify the intravascular components, especially plasma hyaluronan levels, in laparoscopic and conventional colorectal resections that contribute to thromboembolism formation. Methods Prospective cohort analysis of consecutive patients which were participating in the prospective randomized multi-center trial Lapkon II comparing the long-term effects of laparoscopic and conventional resection for colon cancer. Plasma samples were obtained from 15 patients at the beginning and the end of laparoscopic or conventional colorectal resections. Concentrations and activities of tissue plasminogen activator(t-PA), plasminogen activator inhibitor type 1(PAI-1), t-PA/PAI complex, fibrinogen, d-dimers and hyaluronan were determined by using commercial enzyme-linked immunosorbent assay (ELISA) kits. Results No differences in age, sex and type of resection between the laparoscopic and conventional-surgery groups were observed. Laparoscopic procedures lasted longer (p < 0.05). Concentration and activities of t-PA, PAI-1, t-PA/PAI complex, fibrinogen and d-dimers did not vary between the two groups. Plasma hyaluronan decreased from 28.6 to 17.9 IU/ml (p < 0.05) during laparoscopic compared to conventional procedures. Plasma hyaluronan levels were significantly different at the end of operation between the two groups (p < 0.05) . Conclusions Plasma hyaluronan levels were decreased in patients undergoing laparoscopic colorectal resections, compared to those undergoing conventional procedures. Therefore, interactions between plasma hyaluronan and fibrinogen may be lower, with a sequential decrease in fibrin polymerization, and a possibly reduced risk of deep venous thrombosis.     

腹腔镜结直肠癌手术对腹膜微结构损伤和纤维溶解分子表达的影响

目的探讨腹腔镜结直肠癌根治术对腹膜微结构损伤和腹膜纤维溶解分子表达的影响。方法前瞻性纳入2011年6月至2012年2月问山西省人民医院收治的50例结直肠癌患者．根据患者意愿和医生决策分为腹腔镜组（27例）和开腹组（23例）。分别在光镜和电镜下观察术后腹膜微结构损伤程度：采用ELISA法比较手术前后腹膜组织型纤溶酶原激活物t-PA和纤溶酶原激活物抑制因子1（PAI-1）水平的改变。结果术后标本光镜和电镜观察显示,开腹组肠管浆膜完整性受损、系膜脂肪细胞与间皮细胞覆盖脱失、炎性细胞聚集程度较腹腔镜组严重,两组损伤评分比较的总平均秩次分别为38．22和14．67,差异有统计学意义（P〈0．01）。术后网膜组织t-PA、肠管浆膜组织t-PA及PAI-1水平的差异均无统计学意义（均P〉0．05）;但腹腔镜组网膜组织中PAI-1水平显著低于开腹组（P〈0．05）。结论相对于开腹手术,腹腔镜结直肠癌根治术腹膜损伤程度较轻,对腹膜纤溶功能的影响较小．有利于防止肠粘连的形成。     

Influence of the sampling technique on the measurement of peritoneal fibrinolytic activity.

OBJECTIVE: To establish the influence of the peritoneal sampling technique on the measurement of fibrinolytic capacity. DESIGN: Clinical study. SETTING: University hospital, Germany. SUBJECTS: 40 peritoneal biopsy specimens were taken from 10 patients who were having elective colorectal resections. INTERVENTIONS: Peritoneal biopsy specimens were taken either with a biopsy punch (n = 20) or manually with forceps and scissors (n = 20). MAIN OUTCOME MEASURES: Extent of agreement in fibrinolytic activities between specimens taken with biopsy punch and manually. Major endpoint-peritoneal tissue plasminogen activator (t-PA) activity. Minor endpoints-peritoneal tissue plasminogen activator concentration, and concentration and activity of plasminogen activator inhibitior type 1 (PAT-1). RESULTS: Intra-assay agreement and the extent of agreement between the groups were evaluated by the method of Bland and Altman. Correlation of repeated measurements of t-PA and PAI-1 concentrations and activities from the same sample using the same ELISA kit was high (r = 0.93-0.99, p < 0.01). t-PA activities and concentrations between the groups correlated poorly (r= 0.60 and 0.66, p < 0.01) while no correlation at all was seen for PAI-1 concentration and activity between the groups (r = 0.6 and 0.1, p = 0.2 and 0.9). The mean differences between the groups ranged from -27% to -4.8%. CONCLUSION: The sampling technique considerably affects the measurement of peritoneal fibrinolytic activity.     

血清t-PA和PAI-1水平在腹腔镜手术中的临床意义

目的:探讨腹腔镜与开腹手术中血浆纤维蛋白溶酶的变化。方法:在腹腔镜与开腹手术前、术后8h分别测量血浆中组织型纤溶酶原激活物(Tissue-type p lasm inogen activator,t-PA),纤溶酶原激活物抑制物-1(P lasm inogen activator inh ib itortype-1,PAI-1)浓度。结果:血浆t-PA浓度在腹腔镜手术和传统腹部手术两组病例术后均下降,但在传统腹部手术组下降更显著(P<0.05),而血浆PAI-1浓度在腹腔镜手术和传统腹部手术两组病例术后均升高,但在传统腹部手术组升高更显著(P<0.05)。结论:本次临床研究结果表明:腹腔镜手术组术前、术后血浆中t-PA、PAI-1浓度的变化小于传统腹部手术组,腹腔镜手术组引起腹腔粘连严重程度也低于传统腹部手术组。     

Time to reconsider saline as the ideal rinsing solution during abdominal surgery

BACKGROUND: Peritoneal mesothelial cells lining the peritoneal cavity play a primary role in prevention of formation of peritoneal adhesions, which depends mainly on their fibrinolytic activity. During surgical procedures, the abdominal cavity is in most cases rinsed with normal saline solution, which may modify the fibrinolytic activity of the peritoneal mesothelium and predispose to formation of adhesions. The goal of our experiments was to evaluate how normal saline and other solutions affect the fibrinolytic properties of the peritoneal mesothelial cells. MATERIAL AND METHODS: Experiments were performed on in vitro cultures of human peritoneal mesothelial cells. Mesothelial monolayers were exposed during 6 hours to the following solutions: culture medium (control), .9% NaCl, Hanks solution, Earles solution, new peritoneal dialysis fluid with low glucose degradation products (GDP) concentration (PDF), and peritoneal dialysis fluid with high concentration of GDP (PDF-GDP). Afterwards, morphology of the cells as well as leakage of lactate dehydrogenase (LDH) from their cytosol were evaluated. During the next 24 hours when the cells were cultured in standard medium synthesis of interleukin-6 (IL-6), tissue plasminogen activator (t-PA) and plasminogen activator inhibitor-1 (PAI-1) were studied. RESULTS: Mesothelial monolayers exposed to .9% NaCl or to PDF-GDP showed destruction of their morphology after 6 hours incubation and in case of PDF-GDP release of LDH from the cytosol was increased by 275% versus the control (P<.05). During subsequent culture of all cells in standard medium, the release of IL-6 was decreased in cases of cells pretreated with .9% NaCl (-58%, P<.05) or PDF-GDP (-93%, P<.001). The release of t-PA was also reduced from cells pretreated with .9% NaCl (-71%, P<.01) or with PDF-GDP (-74%, P<.01) but increased after exposure of these cells to PDF (+35%, P<.05). Statistically significant decrease of PAI-1 synthesis was observed in cells preexposed to .9% NaCl (-69%, P<.01) or to PDF-GDP (-82%, P<.05). When changes in the PAI-1/t-PA ratio were calculated, a strong tendency for increase of that value was seen in cells pretreated with .9% NaCl or Earles salts solution but not with PDF. However, in cases of Hanks solution, a significant increase in the PAI-1/t-PA ratio was observed (+104%, P<.01). CONCLUSION: Exposure of the peritoneal mesothelial cells to .9% NaCl, Hanks, Earles salts solution, or PDF-GDP results either in reduction of their viability or in loss of their fibrinolytic activity. Peritoneal dialysis fluid with a low content of glucose degradation products appears to be the optimal solution causing the least damage to mesothelial cells and therefore may be the ideal solution for rinsing the abdominal cavity with low risk of inducing deterioration of the mesothelial cells fibrinolytic activity and formation of adhesions. We postulate therefore that such hypertonic peritoneal dialysis fluids should be used not only during peritoneal dialysis but also for rinsing the abdominal cavity during any surgical procedures.     

Peritoneal fibrinolytic activity in peritonitis

BACKGROUND: Peritonitis may cause a reduction in abdominal fibrinolytic activity. The reduced local fibrinolysis seems to be an important process in the subsequent development of adhesion formation. The aim of the study was to evaluate peritoneal fibrinolytic capacity in inflamed and normal peritoneum. METHODS: Peritoneal biopsy specimens were taken at the beginning of operation from 15 patients with peritonitis and 10 patients who underwent elective operation. Levels of tissue-type plasminogen activator (tPA), urokinase-type plasminogen activator (uPA), plasminogen activator inhibitor (PAI) type 1 (PAI-1) and type 2 (PAI-2), and tPA/PAI complex in tissue extracts were determinated by commercially available enzyme-linked immunosorbent assay kits. RESULTS: tPA was significantly reduced in peritonitis compared with normal peritoneum (P <0.001), whereas it was found that the levels of PAI-1, PAI-2, uPA, and tPA/PAI complex in peritonitis were significantly higher than those in normal controls. CONCLUSIONS: Plasminogen activator activity was significantly reduced in peritoneal biopsy samples from patients with peritonitis compared with those from patients without peritonitis.     

Adenovirus-mediated transfer of the 39 kD receptor-associated protein increases fibrinolytic capacity.

BACKGROUND: The mesothelium has an important role in maintaining an adequate fibrinolytic capacity in the peritoneal cavity and thus in preventing the formation of fibrinous peritoneal adhesions by secreting the fibrinolytic enzyme tissue-type plasminogen activator (t-PA). The fibrinolytic activity of human mesothelial cells (HMCs) is counteracted by rapid uptake of t-PA via the low-density lipoprotein receptor-related protein (LRP). The 39 kD receptor-associated protein (RAP) is an inhibitor of binding of t-PA to LRP, but RAP itself is also rapidly degraded via LRP. METHODS: Adenovirus-mediated RAP gene transfer technology was used to evaluate the effect of prolonged overexpression of RAP on t-PA accumulation in conditioned medium of HMCs under basal and inflammatory conditions. RESULTS: Infection of HMCs with a recombinant adenovirus carrying the RAP cDNA resulted within one day in t-PA levels that were maximally twofold to threefold increased as compared with noninfected or adenovirus-beta-galactosidase-infected cells. Whereas upon prolonged incubation, t-PA levels in the conditioned medium of uninfected cells leveled off because of rapid uptake and degradation via LRP, t-PA concentrations in the medium of adenovirus-RAP-infected cells continued to increase, reaching fivefold control levels after 72 hours. The increased t-PA accumulation persisted for seven days and then slowly returned to control values over the next few weeks. In contrast, the production of a specific inhibitor of t-PA, plasminogen activator inhibitor-1 (PAI-1), was not affected by adenoviral RAP gene transfer. Northern blotting analysis showed that t-PA, PAI-1, and LRP mRNA concentrations were not changed after adenoviral infection, underlining that the elevated t-PA levels are the result of RAP-blocked uptake and degradation of t-PA rather than increased t-PA synthesis. RAP gene transfer also restored diminished fibrinolytic activity of cytokine-treated mesothelial cells. CONCLUSIONS: Adenovirus-mediated transfer of the RAP gene provides an efficient way of transiently increasing the fibrinolytic capacity of mesothelial cells.     

Decreased fibrinolytic capacity in coronary patients by increased plasminogen activator inhibitor activity]

The fibrinolytic capacity of the blood mainly depends on the amount of tissue-plasminogen activator (t-PA) antigen and plasminogen activator inhibitor (PAI). In this study the fibrinolytic response to a venous occlusion test (VOT) was measured in 109 patients with angiographically documented coronary artery disease (CAD) and in 20 healthy volunteers at comparable age (controls). CAD-patients had higher plasma plasminogen activator inhibitor capacity before (24.4 +/- 11.0 vs. 15.4 +/- 5.2 arbitrary units [AU/ml]; p less than 0.0002) and after VOT (19.6 +/- 13.2 vs. 10.9 +/- 5.3 AU/ml; p less than 0.0001) compared with controls. Furthermore they showed significant lower plasma t-PA activity after VOT (3.0 +/- 6.8 vs. 6.6 +/- 10.6 AU/ml; p less than 0.0001). However there were no difference between both groups in plasma t-PA antigen levels after VOT (17.3 +/- 12.1 vs. 18.7 +/- 14.4 ng/ml). In 10% of patients the decrease in fibrinolytic activity resulted from a lower t-PA release ("lower" was defined as mean minus one standard deviation of the control group). 40% showed elevated plasma PAI capacity before VOT ("elevated" was defined as mean plus two standard deviations of the control group). Both caused significantly reduced post occlusion plasma t-PA activity and prolonged Euglobulin clot lysis time (p less than 0.003). A positive correlation was found between PAI capacity and serum triglyceride levels. Reduced fibrinolytic activity in 109 patients with coronary heart disease based either on a decrease in t-PA antigen release or a increased in PAI capacity in comparison with healthy controls. The mechanism of these findings is not yet well-known.(ABSTRACT TRUNCATED AT 250 WORDS)     

Simvastatin increases fibrinolytic activity in human peritoneal mesothelial cells independent of cholesterol lowering

BACKGROUND: The continuous physical and chemical irritation of the peritoneum in peritoneal dialysis patients can result in a nonbacterial serositis with increased fibrin deposition, thus promoting peritoneal fibrosis and adhesion development. By expressing the fibrinolytic enzyme tissue-type plasminogen activator (t-PA) and its specific inhibitor, plasminogen activator inhibitor-1 (PAI-1), human peritoneal mesothelial cells (HMC) play an important role in regulating peritoneal fibrinolysis. METHODS: Cultured HMC were used to examine the effect of a 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitor, simvastatin, on the expression of t-PA and PAI-1. Antigen concentrations in the cell supernatants were measured by ELISA and Northern blot analysis was conducted for mRNA expression. RESULTS: Simvastatin time- and concentration-dependently increased t-PA and decreased PAI-1 synthesis, reaching maximal effects after 48 hours, when simvastatin (1 micromol/L) increased t-PA levels 5.1 +/- 0.1-fold and suppressed PAI-1 levels 2.6 +/- 0.2-fold. This was accompanied by a twofold increase in mesothelial cell-associated t-PA activity. Qualitatively similar results were obtained in cultured human endothelial cells, but the effects were less pronounced and required higher simvastatin concentrations. Northern blot analysis revealed that the action of simvastatin on t-PA and PAI-1 expression in HMC can be explained by parallel changes in t-PA and PAI-1 mRNA. The effects of simvastatin were prevented in the presence of mevalonate and geranylgeraniol, suggesting that the effect of simvastatin on t-PA and PAI-1 synthesis is mediated through geranylgeranyl-modified intermediates. Experiments using specific inhibitors of geranylgeranylated Rho GTPases excluded a role of members of this family of small GTP-binding proteins in simvastatin action in HMC. The effects of simvastatin on t-PA and PAI-1 expression as well as on cell shape were completely mimicked by cytochalasin D, a disrupter of cellular actin filaments, but not by colchicine, a disrupter of microtubules. CONCLUSIONS: In conclusion, the cholesterol-lowering drug simvastatin is an effective stimulator of local peritoneal fibrinolytic activity, as it increases t-PA and decreases PAI-1 production in mesothelial cells by a mechanism involving geranylgeranyl-modified intermediates and actin skeleton perturbation. These results provide a new rationale to prevent peritoneal fibrin deposition and adhesion development in peritoneal dialysis patients.     

Fibrinolytic capacity in hemodialysis patients treated with recombinant human erythropoietin.

A major adverse effect of recombinant human erythropoietin (r-HuEPO) in hemodialyzed patients are thrombotic events. Several reports on platelet function during r-HuEPO treatment have been published but less is known about fibrinolysis. In the present study, the fibrinolytic capacity was studied in 20 patients on maintenance hemodialysis and treated with r-HuEPO. The patients were randomized into two groups and investigated in a crossover design. r-HuEPO was administered intravenously and subcutaneously in each group and was given for 3 months, respectively. Plasma tissue plasminogen activator (t-PA) and released t-PA remained unaffected by r-HuEPO in both groups throughout the study. Tissue plasminogen activator inhibitor (PAI) increased in a cyclic way reaching peak values 4-6 weeks after the start of investigation and again 4-6 weeks after changing therapy. The increase in PAI was significant in the two groups (0.025 > p > 0.01). Tissue plasminogen antigen was low in the uremic patients. The influence of r-HuEPO on this parameter was not investigated. Compensatory changes in plasma levels of factor XII procoagulant activity, activated protein C and of alpha 2-antiplasmin were not observed. Thrombotic events occurred in 4 patients at peak values of PAI. Six patients required an increase in heparin dose simultaneously with the increase in PAI. Thus, r-HuEPO seemed to affect the fibrinolytic capacity of uremic patients.     

Effects of seprafilm on peritoneal fibrinolytic system

BACKGROUND: There is a high incidence of adhesions after ventral hernia repair with polypropylene mesh. The purpose of the present study was to evaluate the efficacy of Seprafilm in the prevention of adhesion formation and effect on peritoneal fibrinolytic activity. METHODS: An incisional hernia model was created in rats. In the experimental group Seprafilm was placed between polypropylene mesh and abdominal organs. On the 14th day adhesions were evaluated and tissue plasminogen activator (tPA), urokinase plasminogen activator (uPA), plasminogen activator inhibitor (PAI) type 1 and 2 were measured in peritoneal biopsy specimens. Results: Adhesions were significantly reduced in the Seprafilm group (P = 0.002). Nevertheless, there were no difference between the two groups in levels of tPA, PAI-1 and PAI-2. However, the levels of uPA were significantly decreased in the Seprafilm group. CONCLUSIONS: The adhesion preventive effect of Seprafilm is not directly related in peritoneal fibrinolytic activity. Instead, the physical properties (barrier, hydroflotation and sliconizing effect) of the membrane are primarily responsible for adhesion prevention.     

Peritoneal response to pneumoperitoneum and laparoscopic surgery

BACKGROUND: It is generally believed that laparoscopic surgery inflicts less trauma to the peritoneum than open surgery. Local peritoneal fibrinolysis is a critical factor in adhesion development. The objective was to investigate fibrinolytic changes in the peritoneum during laparoscopic and open surgery. METHODS: At laparotomy (n = 10) peritoneal biopsies were taken at opening of the abdomen and just before closure. At laparoscopy (n = 12) opening peritoneal biopsies were taken after carbon dioxide insufflation, and closure biopsies just before exsufflation. Tissue concentrations of tissue-type plasminogen activator (tPA), plasminogen activator inhibitor type 1 (PAI-1) and the resulting tPA activity were assayed. RESULTS: Concentrations of tPA in peritoneal tissue declined during operation in both groups, but significantly so only in the laparotomy group (- 53 per cent; P = 0.01). PAI-1 levels were higher in opening biopsies from the laparoscopy group (P = 0.004). There was an increase in PAI-1 concentration during laparotomy, but not during laparoscopy. At the end of the operation, there was no difference between the groups. The resulting tPA activity did not differ between groups at opening or closure. In both groups there was a significant decline during operation (laparotomy: - 59 per cent, P = 0.02; laparoscopy: - 63 per cent, P = 0.01). CONCLUSION: These findings indicate that the peritoneal response to open and laparoscopic surgery is similar. The initial rise in peritoneal PAI-1 concentration during laparoscopy suggests an adverse effect of carbon dioxide insufflation, which might affect peritoneal repair.     

Tissue-type plasminogen activator activity in HIV-associated HUS

Background: In children, haemolytic uraemic syndrome (HUS) is associated with high plasma plasminogen activator inhibitor type 1 (PAI-1), which may contribute to the persistence of renal glomerular and arteriolar thrombi. HUS has been described in HIV-infected patients, but the pathophysiology of HUS in these patients is poorly understood. The aim of the study was to investigate plasma fibrinolytic activity in 18 patients with HIV-associated HUS. Methods: We measured tissue type plasminogen activator (t-PA) and PAI-1 activities in the plasma of 18 HIV-infected patients with biopsy-proven HUS (HIV+/HUS+0 and 48 HIV-infected patients without HUS (HIV+/HUS-). Results: Patients with HUS had a significantly higher serum creatinine, a lower platelet count and an increased incidence of cytomegalovirus (CMV) infection (72% of patients HIV+/HUS+, vs 25% of patients HIV+/HUS-). Unexpectedly, plasma PAI-1 activity was similar in both groups. However, t-PA activity was significantly higher in HUS cases (11.5 vs 4.5 U/ml, P=0.001). Patients with CMV infection, with or without HUS, had significantly increased t-PA level (P=0.01). Multivariate analysis identified high t-PA (RR=9.21) and CMV infection (RR=3.36) as risk factors for HUS. Conclusion: This study provides evidence that HIV-infected patients with HUS have high plasma t-PA activity. PAI-1 plasma activity is not significantly increased, as opposed to non-HIV-associated HUS. Thus, in the setting of HIV infection, HUS cannot be attributed to decreased fibrinolytic activity.     

Laparoscopic-type environment enhances mesothelial cell fibrinolytic activity in vitro via a down-regulation of plasminogen activator inhibitor-1 activity

BACKGROUND: Fewer intraperitoneal adhesions have been observed after laparoscopic surgery compared with conventional techniques. The aim of this study is to assess the effect of the pneumoperitoneum on mesothelial cell fibrinolytic activity by use of an in vitro model. METHODS: Human peritoneal mesothelial cells were seeded onto 24-well plates and incubated in carbon dioxide or helium at 5 mm Hg for 4 hours or standard culture conditions. Supernatant was removed for analysis at 0, 24, 48, and 72 hours after gas incubation and analyzed for plasminogen activator activity, total tissue plasminogen activator (tPA), and total plasminogen activator inhibitor-1 (PAI-1) concentrations by use of an enzyme-linked immunosorbent assay. The effect of different insufflation pressures (0, 7, and 14 mm Hg) was also examined. RESULTS: Enhanced plasminogen activator activity was observed at 48 hours and 72 hours from cells exposed to CO(2) (P<.04 each) and helium (P<.05 each) compared with control. This was associated with a decrease in PAI-1 concentrations at 48 and 72 hours in both the CO(2) and helium groups compared with control (P<.03 each, CO(2) vs control; and P<.04 each, helium vs control). No changes in tPA levels were observed. Changes in insufflation pressures did not affect plasminogen activator activity. CONCLUSIONS: These results suggest that incubation of human mesothelial cells with both CO(2) and helium in the absence of oxygen enhances mesothelial cell fibrinolytic activity because of a reduction in PAI-1 concentrations. These changes may participate in the observed reduction in adhesions after laparoscopic surgery relative to open surgery.     

Secretory activity of the coronary artery endothelial cells in conditions of the peritoneal dialysis

IntroductionEndothelial dysfunction is frequent in patients treated with peritoneal dialysis and may lead to cardiac complications. We evaluated the effect of effluent dialysates and serum on the function of coronary artery endothelial cells (CAEC).MethodsHuman CAEC in in vitro culture were exposed to serum and dialysates from 24 patients treated with continuous ambulatory peritoneal dialysis (CAPD) and secretion of interleukin-6 (IL6), von Willebrand factor (vWF), tissue plasminogen activator (t-PA) and plasminogen activator inhibitor-1 (PAI-1) were measured. Modulation of the secretory activity of CAEC by Sulodexide, mixture of glycosaminoglycans: heparin sulfate and dermatan sulfate, was studied.ResultsSerum from CAPD patients stimulated synthesis of IL6 (+93%), vWF (+18%), and PAI-1 (+20%) and did not change t-PA secretion in CAEC. Dialysates stimulated secretion of IL6 (+89%), vWF (+29%), and PAI-1 (+31%) and did not change t-PA synthesis. Dialysates collected in 12 patients after 6 months more strongly stimulated synthesis of IL6 (+37%) and PAI-1 (+7%). Sulodexide suppressed the secretory activity of CAEC stimulated by the studied sera: IL6 (–38%), vWF (–19%), t-PA (–13%), and PAI-1 (–12%).ConclusionsSerum and the dialysate from CAPD patients induce inflammatory and prothrombotic reaction in coronary arterial endothelial cells. The general pattern of the observed effects for serum and dialysates was similar but the intensity of the effects was not identical. Sulodexide reduced these effects.     

Intraperitoneal hypercoagulation and hypofibrinolysis is present in childhood peritonitis

 An increased rate of obstruction of peritoneal dialysis catheters is observed during peritonitis. Hypercoagulation and hypofibrinolysis may explain this increased occurrence. We studied plasminogen activator inhibitor type 1 antigen (PAI-1), tissue-type plasminogen activator antigen (t-PA), D-dimer (DD), plasmin-α₂-antiplasmin complexes (PAP), and thrombin-antithrombin III complexes (TAT) in 7 children with peritonitis (group A) and 12 children during stable peritoneal dialysis (group B). Albumin, β₂-microglobulin, IgG, and α₂-macroglobulin were measured for baseline transperitoneal protein transport. After a dwell of 6 h with 1.36% Dianeal, dialysate and serum samples were collected. Dialysate to plasma ratios of all proteins were calculated. During peritonitis (group A) TAT was higher: 34.7 versus 22.0 (P=0.01). PAI-1 was increased in group A: 76.5 versus 22.9 (P=0.004). PAP was decreased during peritonitis (group A): 24.9 versus 39.3 (P=0.01). In group A, DD were decreased. 10.8 versus 26.7 (P=0.002). t-PA was similar in both groups (23.7 in group A vs. 27.7 in group B; P=0.26). In both groups TAT, PAI-1, t-PA, PAP, and DD were significantly higher than in baseline transperitoneal transport, suggesting intraperitoneal production. Hypercoagulability and hypofibrinolysis were present during peritonitis compared with the control situation. Received: 9 April 1998 / Revised: 11 August 1998 / Accepted: 12 August 1998