Plasma PAI-1 Research Articles

Abstract 1367: Anti-angiogenesis promotes venous thromboembolism through inducing PAI-1 in a mouse xenograft model of human lung carcinoma

Abstract Thromboembolism is a major source of morbidity and mortality in patients with cancer. Increased incidence of venous thromboembolism (VTE) was associated with anti-vascular endothelial growth factor (VEGF) treatment in cancer. However, the mechanism of VTE initiation remains elusive. In this study, we addressed the effect of bevacizumab, a humanized monoclonal antibody against VEGF-A on either an inferior vena cava stenosis or a saphenous vein model using a xenograft mouse model. We found that treatment of bevacizumab accelerated thrombosis in a saphenous vein model with an increased thrombus weight in an inferior vena cava stenosis model. Administration of bevacizumab inhibited the growth of lung tumor by suppressing angiogenesis. Both intrathrombotic PAI-1 expression and plasma PAI-1 level was significantly increased by the treatment of bevacizumab. Furthermore, pharmacologic inhibition of PAI-1 with PAI-039 blocked the bevacizumab-induced venous thrombosis, but showed no change in tumor growth. Collectively, these findings indicate that PAI-1 plays a role in VET with anti-angiogenesis therapy, and have efficacy as a therapeutic strategy to prevent VET. Citation Format: Ni Chen, Meiping Ren, Rong Li, Xin Deng, Yongjie Li, Kai Yan, Lamei Xiao, Yan Yang, Liqun Wang, Mao Luo, Jianbo Wu. Anti-angiogenesis promotes venous thromboembolism through inducing PAI-1 in a mouse xenograft model of human lung carcinoma. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 1367. doi:10.1158/1538-7445.AM2015-1367

Evaluating PAI-1 as a biomarker for stress in diving: human serum total PAI-1 is unaltered after 2 h dry exposures to 280 kPa hyperbaric air

Plasminogen activator inhibitor (PAI-1) is induced in the vasculature and secreted into the vascular lumen in response to inflammation and oxidative stress. We have previously reported a fivefold increase in plasma PAI-1 from rats exposed to 708 kPa hyperbaric air. In the current study we assess the potential of human serum total PAI-1 as a biomarker for stress in compressed air diving. Eleven recreational divers, nine males and two females, completed four 2 h hyperbaric air exposures to 280 kPa in a pressure chamber over a period of 2 weeks. The air pressure corresponds to a diving depth of 18 m in water. Serum was collected before the study and again 3 h 30 min after completion of each hyperbaric exposure. All samples were taken in the afternoon to minimize the contribution of circadian variation. The analysis revealed no change in serum total PAI-1 after hyperbaric exposures within the group of divers (P = 0.064), but significant interindividual differences persisted throughout the study (P < 0.0005). A case of decompression sickness after the third round of hyperbaric exposure did not affect PAI-1. In conclusion, compressed air exposure to 280 kPa does not affect serum total PAI-1, and significant interindividual variation in PAI-1 levels may limit its usefulness as a biomarker. This does, however, not give a complete answer regarding PAI-1 in physiologically stressful dives. Further studies with different exposures and timing are needed for that.

Effect of Grape Consumption on Biomarkers of Endothelial and Cardiovascular Health

Polyphenols have been associated with cardio‐protective benefits, however causal mechanisms remain elusive. This randomized, double‐blind, crossover study examines acute and longer‐term effects of grape consumption on biomarkers of endothelial and cardiovascular health. It was hypothesized that polyphenol‐rich grape consumption would protect against diet‐induced elevations and long‐term impairment in biomarkers of endothelial dysfunction during the acute postprandial period and after 4 weeks respectively. Twenty obese (BMI 37.0±9.9) but otherwise healthy adults were provided with isocaloric sachets of freeze‐dried whole grape powder (grape) (1 60g sachet/day=297mg polyphenol) and placebo (1 60g sachet/day=0mg polyphenol) for daily consumption, each treatment for 4 weeks separated by 2 week washout. To examine acute effects, postprandial blood samples were collected at 0, 1, 3 and 5 hours following a high fat high carbohydrate (HFHC) meal challenge and grape or placebo consumption. Longer‐term effects were examined by fasted blood samples collected on the last day of each 4‐week intervention. HDL increased (p&lt;0.05) after 4 week grape consumption, but no other significant changes in lipids or blood glucose were noted between groups. Data analyzed by repeated measures, 2‐way ANOVA, showed that PAI‐1 and MCP‐1 were significantly elevated in plasma after HFHC challenge (p&lt;0.05). However, no significant differences in plasma sICAM, sVCAM, ET‐1, PAI‐1 or MCP‐1 were detected between grape and placebo treatments after HFHC challenge or 4‐week intervention. Grape consumption has some beneficial effects on markers of cardiovascular health.Supported by California Table Grape Commission and USDA CRIS CA‐S*‐NTR‐6316‐H

Effects of a High-Fat Diet on Spontaneous Metastasis of Lewis Lung Carcinoma in Plasminogen Activator Inhibitor-1 Deficient and Wild-Type Mice

This study investigated the effects of a high-fat diet on spontaneous metastasis of Lewis lung carcinoma (LLC) in plasminogen activator inhibitor-1 deficient (PAI-1−/−) and wild-type mice. The high-fat diet increased the number of pulmonary metastases by 60% (p<0.01), tumor cross-sectional area by 82% (p<0.05) and tumor volume by 130% (p<0.05) compared to the AIN93G diet. Deficiency in PAI-1 reduced the number of metastases by 35% (p<0.01) compared to wild-type mice. In mice fed the high-fat diet, PAI-1 deficiency reduced tumor cross-sectional area by 52% (p<0.05) and tumor volume by 61% (p<0.05) compared to their wild-type counterparts; however, PAI-1 deficiency affected neither area nor volume in mice fed the AIN93G diet. Adipose and plasma concentrations of PAI-1 were significantly higher in high-fat fed wild-type mice than in their AIN93G-fed counterparts. Adipose and plasma PAI-1 were not detectable in PAI-1−/− mice regardless of the diet. Mice deficient in PAI-1 showed significantly greater plasma concentrations of monocyte chemotactic protein-1, tumor necrosis factor-α, leptin, vascular endothelial growth factor, tissue inhibitor of metalloproteinase-1 and insulin compared to wild-type mice, indicating a compensatory overproduction of inflammatory cytokines, angiogenic factors and insulin in the absence of PAI-1. We conclude that PAI-1 produced by the host, including that by adipose tissue, promotes high-fat enhanced metastasis of LLC.

Profibrinolytic effect of the epigenetic modifier valproic acid in man.

AimsThe aim of the study was to test if pharmacological intervention by valproic acid (VPA) treatment can modulate the fibrinolytic system in man, by means of increased acute release capacity of tissue plasminogen activator (t-PA) as well as an altered t-PA/Plasminogen activator inhibitor -1 (PAI-1) balance. Recent data from in vitro research demonstrate that the fibrinolytic system is epigenetically regulated mainly by histone deacetylase (HDAC) inhibitors. HDAC inhibitors, including VPA markedly upregulate t-PA gene expression in vitro.Methods and ResultsThe trial had a cross-over design where healthy men (n = 10), were treated with VPA (Ergenyl Retard) 500 mg depot tablets twice daily for 2 weeks. Capacity for stimulated t-PA release was assessed in the perfused-forearm model using intra-brachial Substance P infusion and venous occlusion plethysmography. Each subject was investigated twice, untreated and after VPA treatment, with 5 weeks wash-out in-between. VPA treatment resulted in considerably decreased levels of circulating PAI-1 antigen from 22.2 (4.6) to 10.8 (2.1) ng/ml (p<0.05). It slightly decreased the levels of circulating venous t-PA antigen (p<0.05), and the t-PA:PAI-1 antigen ratio increased (p<0.01). Substance P infusion resulted in an increase in forearm blood flow (FBF) on both occasions (p<0.0001 for both). The acute t-PA release in response to Substance P was not affected by VPA (p = ns).ConclusionValproic acid treatment lowers plasma PAI-1 antigen levels and changes the fibrinolytic balance measured as t-PA/PAI-1 ratio in a profibrinolytic direction. This may in part explain the reduction in incidence of myocardial infarctions by VPA treatment observed in recent pharmacoepidemiological studies.Trial RegistrationThe EU Clinical Trials Register 2009-011723-31

GW25-e0783 Association between Plasma PAI-1 Levels and Clock Genes Polymorphisms in Primary Hypertensions

GW25-e0783 Association between Plasma PAI-1 Levels and Clock Genes Polymorphisms in Primary Hypertensions

Abstract 245: Consumption of a high-fat diet abrogates inhibitory effects of selenium on spontaneous metastasis of Lewis lung carcinoma in mice

Abstract The present study investigated the effects of dietary supplementation with selenium (Se) on spontaneous metastasis of Lewis lung carcinoma (LLC) in mice fed a high-fat diet using a 2x2 design. Three-week-old male C57BL/6 mice were fed the AIN93G diet or that diet modified with 45% calories from fat supplemented with or without 2.5 mg Se/4029 kcal as methylseleninic acid for 6 weeks, at which time they were injected subcutaneously with 2.5x105 LLC cells. The resulting primary tumor was removed surgically 10 days later, and the experiment was terminated after an additional 10 days. Consumption of the high-fat diet significantly increased the body fat mass, regardless of Se supplementation, compared to the AIN93G diet (p&amp;lt;0.01). The high-fat diet increased lung metastases by 17% compared to the AIN93G diet (p&amp;lt;0.01). Selenium supplementation reduced the metastases by 11% compared to non-supplemented controls (p&amp;lt;0.05); however, the reduction was greater in AIN93G-fed mice (18%) than in mice fed the high-fat diet (5%). Supplemental Se reduced plasma concentrations of proteolytic proteases (uPA, p&amp;lt;0.01; MMP-9, p&amp;lt;0.05) and angiogenic factors (VEGF, p&amp;lt;0.01; TIMP-1, p&amp;lt;0.01) compared to non-supplemented controls. The high-fat diet significantly increased plasma concentrations of adipokines PAI-1, MCP-1, TNF-α and leptin regardless of the level of dietary Se; however, supplemental Se reduced plasma PAI-1 (p≤0.05) and MCP-1 (p≤0.05) in AIN93G-fed mice. These results demonstrate that consumption of a high-fat diet abrogated the anti-metastatic effects of Se, and that such an abrogation may involve adipose-derived inflammatory cytokines. Citation Format: Lin Yan. Consumption of a high-fat diet abrogates inhibitory effects of selenium on spontaneous metastasis of Lewis lung carcinoma in mice. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 245. doi:10.1158/1538-7445.AM2014-245

Effects of Saskatoon berry powder on monocyte adhesion to vascular wall of leptin receptor-deficient diabetic mice

Hypothesis:Atherosclerotic cardiovascular complications are the leading cause of death in diabetic patients. Monocyte adhesion is an early event for atherogenesis. Previous studies demonstrated that dark-skin berries had cardiovascular protective effects. We hypothesize that Saskatoon berry (SB) powder may reduce monocyte adhesion in leptin receptor-deficient (db/db) diabetic mice. MethodsWild-type and db/db mice were fed with chow or supplemented with SB powder. Anthocyanins in SB powder were identified using mass spectrometry. Mouse monocytes were incubated with mouse aorta. Monocyte adhesion was counted under microscopy. Inflammatory or metabolic markers in blood or tissue were analyzed using immunological or biochemical methods. ResultsSB powder significantly reduced monocyte adhesion to aorta from diabetic db/db mice compared to regular chow. The increased monocyte adhesion to aorta was normalized in db/db mice treated with ≥5% of SB powder for 4weeks. Increased contents of Nicotinamide adenine dinucleotide phosphate oxidase (NADPH) oxidase-4, heat shock factor-1, monocyte chemotactic protein (MCP)-1, intracellular adhesion molecule (ICAM)-1, P-selectin, tumor necrosis factor-α, plasminogen activator inhibitor (PAI)-1 and urokinase plasminogen activator in aorta or heart apex, elevated plasma PAI-1 and MCP-1 were detected in db/db mice on chow compared to wild-type mice on the same diet; 5% SB powder inhibited the increases of inflammatory, fibrinolytic or stress regulators in aorta or heart apex of db/db mice. Monocyte adhesion positively correlated with blood glucose, cholesterol, body weight, heart MCP-1, PAI-1 or ICAM-1. ConclusionThe findings suggest that SB powder attenuated monocyte adhesion to aorta of db/db mice, which was potentially mediated through inhibiting the inflammatory, stress and/or fibrinolyic regulators.

Factors Associated with Early Platelet Activation in Obese Children

To investigate the factors associated with platelet activation in obese children. Cross-sectional study. Department of Pediatrics of Regional Hospital N∘ 1 of Mexican Institute of Social Security in Morelia, Michoacán, Mexico. 79 obese and 64 non-obese children between the ages of 5 and 10 years. Obese children (body mass index [BMI] >85 in growth curves for Centers for Disease Control/National Center for Health Statistics), and the control group of 64 non-obese children (percentile <85), % body fat, platelet activation was assessed by sP-selectin. Other measures were leptin, uric acid (UA), von Willebrand Factor (vWF), plasminogen activator inhibitor (PAI-1), lipid profile, and glucose. Obese children displayed higher plasma sP-selectin, leptin, PAI-1, and vWF than non-obese children. In the univariate logistic regression analysis, leptin, vWF, UA, and high density lipoprotein (HDL), but not with PAI-1, were factors associated with platelet activation. By stepwise linear regression analysis adjusted by sex and age, the best predictor variables for platelet activation were leptin (β:0.381; t:4.665; P=0.0001), vWF (β:0.211; t:2.926; P=0.004), UA (β:0.166; t:2.146; P=0.034), and HDL (β:-0.215; t:-2.819; P=0.006). Obese children have a higher risk of developing early platelet activation. Factors associated with platelet activation were Leptin, vWF, UA, and HDL. Further studies involving larger numbers of patients over a longer duration are needed to understand the possible molecular mechanism underlying the association between leptin, vWF, and UA and endothelial activation and/or endothelial damage/dysfunction in obese children and its influence in cardiovascular disease in adults.

Changes of plasminogen activator inhibitor-1 and D-dimer during continuous blood purification and related factors

To observe the changes of plasminogen activator inhibitor-1 and D-dimer during continuous blood purification (CBP) and related factors. Sixteen patients who were diagnosed with multiple organ dysfunction syndrome (MODS) were randomly divided into 2 groups: 8 patients received standard continuous blood purification with heparin anticoagulation, and the other 8 received CBP without anticoagulation. Ten normal blood samples were collected from healthy volunteers as controls. All patients underwent CBP for 8 h. Blood was taken from those patients at 0, 15, 60, 120 and 480 min during the CBP. Plasma plasminogen activator inhibitor-1, D-dimer and serum TNF-α and IL-1β were measured by ELISA. Plasma levels of PAI-1 and D-dimer were increased significantly compared with those in the control group (P<0.05). Plasma level of PAI-1 was reduced (P<0.05) and D-dimer was increased (P<0.05) after the CBP. The level of plasma PAI-1 in the heparin group was significant reduced compared with the group of CBP without anticoagulation (P<0.05). There was negative correlation between the level of PAI-1 and the dosage of heparin used during a CBP session in the heparin group (r=-0.746, P<0.001). The level of PAI-1 and D-dimer is higher in patients with MODS than that in the normal controls. After the CBP treatment, there is significant decrease in PAI-1 and increase in D-dimer in both groups. Heparin used during CBP can reduce PAI-1 which intensifies its function of anticoagulation.

Abstract P1-04-02: Correlation between blood markers of hemostasis and circulating tumor cells (CTCs) in early breast cancer patients

Abstract Background: Cancer is a risk factor for venous thromboembolism (VTE) and plasma D-dimer (DD) and tissue factor (TF) are established blood markers associated with VTE. CTCs are an independent predictor of survival in early and metastatic breast cancer (BC) patients (pts) and CTCs are associated with the risk of VTE in metastatic BC pts. In this study, we hypothesized correlation between plasma DD and TF with presence of CTCs. Moreover, we hypothesized, that activation of urokinase plasminogen activator system (uPA) could be involved in CTCs released into peripheral blood (PB) as well as coagulation activation. Methods: This prospective study included 119 early BC patients treated by primary surgery followed by systemic therapies when indicated, at the NCI in Slovakia from March to December 2012. Isolated peripheral blood mononuclear cells (PBMC) were depleted of hematopoietic cells (CD45+) using RossetteSepTM negative selection kit. RNA extracted from CD45-depleted PBMC was interrogated for expression of EMT-inducing transcription factors (TWIST1, SNAIL1, SLUG, ZEB1) and epithelial (CK19) gene transcripts by qRT-PCR. Expressions of gene transcripts in CD45- PBMC of pts and healthy donors (HD) were compared. Patient samples with higher epithelial and/or mesenchymal gene transcripts than those of HD (n = 60) were considered as CTC positive. Plasma DD, TF, uPA (urokinase-type plasminogen activator) and PAI-1 (plasminogen activator inhibitor) levels were detected by ELISA, while expressions of TF and uPA system in surgical specimens were evaluated by immunohistochemistry. Results: CTCs were detected in 27.7% pts. The majority of CTCs exhibited either epithelial differentiation (13.5% pts) or mesenchymal phenotype (EMT markers in 15.1% of pts). A small fraction of CTCs showed co-expression of both markers (0.8%). Pts with any CTCs in PB had significantly higher mean ± SEM plasma DD levels (ng/mL) than those of pts without CTCs (632.4 ± 75.9 vs. 365.4 ± 47.0, p = 0.00002). This association was observed for both, epithelial CTCs (688.4 ± 110.4 vs. 400.7 ± 43.5 (p = 0.026) and mesenchymal CTCs (573.1 ± 105.8 vs. 415.6 ± 44.7, p = 0.0007). There was no association between plasma TF levels or breast tumor TF expression and CTCs. Patients with any CTCs had higher mean ± SEM plasma uPA (ng/mL) (374.6 ± 32.6 vs. 307.1 ± 20.3, p = 0.02) and PAI-1 (pg/mL) levels (5.3 ± 0.7 vs. 4.4 ± 0.4, p = 0.28), than those of patients without CTCs, however, there was no correlation between plasma uPA system activation and DD or TF levels. In multivariate analysis CTCs, patients age and tumor grade were independent factors associated with plasma DD levels. Conclusion: This prospective study showed for the first time a positive association between plasma D-dimer levels and CTCs indicating a potential role for activation of the coagulation cascade in the early metastatic process. We hypothesize that CTCs could be directly involved in coagulation activation in breast cancer patients, or alternatively an increased CTCs count could be a marker of more aggressive disease and increased risk of VTE. Future studies will need to address the prognostic implications of these observations with the potential for therapeutic interventions for the metastatic process. Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P1-04-02.

Functionally stable plasminogen activator inhibitor-1 in a family with cardiovascular disease and vitiligo

Vitiligo is a common skin condition with a complex pathophysiology characterized by the lack of pigmentation due to melanocyte degeneration. In this study, we investigated PAI-1 antigen (Ag) and activity levels in a 34 year old male with extensive vascular disease, alopecia areata and vitiligo. Fasting PAI-1 Ag and activity levels were measured at 9 a.m. in the subject and family members. Both PAI-1 Ag (67 ± 38 vs. 18.6 ± 6.5 ng/ml, P < 0.001) and specific activity (15.8 ± 10.0 vs. 7.6 ± 6.0 IU/pmol, P < 0.04) levels of PAI-1 were moderately elevated in subjects compared to the controls. PAI-1 kinetic studies demonstrated a markedly enhanced stability of plasma PAI-1 activity in the family members. Specific activity at 16 h was significantly higher than expected activity levels (0.078 ± 0.072 vs. 0.001 ± 0.001 IU/ng/ml, P < 0.001). While the exact mechanism of increased stability of PAI-1 activity in vitiligo is not known, it is likely due to post-translational modifications or increased binding affinity for a stabilizing cofactor. In conclusion, enhanced stability of PAI-1 may contribute to the pathophysiology of vascular disease and associated melanocyte degeneration. Systemic or local treatment with PAI-1 inhibitors may offer a potential treatment alternative to the near orphan status for vitiligo drug development.

Peptidergic regulation of plasminogen activator inhibitor‐1 gene expression in vivo

The mechanisms by which PAI-1 biosynthesis is altered during stress have not been fully elucidated. Studies suggest a major role for neuro-peptidergic modulation of the stress response by PACAP (pituitary adenylate cyclase-activating polypeptide), a member of the VIP/secretin/glucagon family. We tested the hypothesis that PACAP regulates PAI-1 biosynthesis during stress in vivo. PAI-1 gene expression was monitored by RT-PCR in adrenal glands harvested from C57BL/6J mice that were unstressed, or subjected to restraint stress for 2h, or treated with PACAP. PAI-1 mRNA expression was markedly increased in adrenals from stressed mice. Restraint stress resulted in much smaller increments in adrenal tPA mRNA, suggesting that local adrenal tPA/PAI-1 biosynthetic balance is markedly altered by stress. The observed increases in PAI-1mRNA during stress were substantially blunted (55±4%, P<0.001) by pretreatment with the specific PACAP receptor antagonist, PACAP6-38, compared with pretreatment with vehicle. Administration of the agonist PACAP1-38 alone resulted in a dose-dependent increase in tissue PAI-1 mRNA. PACAP1-38 administration also resulted in substantial increases in plasma PAI-1 antigen and active PAI-1 concentrations that were significantly greater in male mice than in female mice. We conclude that adrenal PAI-1 mRNA expression is markedly increased by stress, and that the PACAP peptidergic signaling pathway plays a major role in mediating the stress-induced increase in PAI-1 biosynthesis.

Effects of Parecoxib on The Prevention of Postoperative Peritoneal Adhesions in Rats

ABSTRACTBackground: No systemic preventive therapy has been successful in inhibiting the development of postoperative peritoneal adhesions (PPAs). Objective: The aim of this study was to evaluate the potential effects of 5 day administration of parecoxib, on PPA prevention and on suture or wound healing in rats. Methods: In a model of PPAs induced by peritoneal electrical burn, 30 rats were randomized into 3 groups according to parecoxib administration route (control; intraperitoneal (IP); intramuscular (IM)). Plasma and peritoneal levels of PAI-1 and tPA were measured at T0, after 90 min of surgery (T90), and on postoperative day 10 (D10). In a cecum resection model, 20 rats were randomized into two groups (control and IP parecoxib), and abdominal wound healing and suture leakage were assessed at D10. In both models, PPAs were evaluated quantitatively and qualitatively on D10. Results: Administration of parecoxib significantly decreased the quantity (p < .05) and the severity (p < .01) of PPAs in both models. In addition, parecoxib administration did not cause healing defects or infectious complications in the two models. In the peritoneal burn model, IP or IM parecoxib administration inhibited the increase of postoperative plasma and peritoneum PAI-1 levels, an increase that was observed in the control group (p < .01). No anastomosis leakage could be demonstrated in both groups in the cecum resection model. Conclusion: This study showed that, in these rat models, parecoxib might reduce PPA formation. Confirmation of the safety of parecoxib on intestinal anastomoses is required and should be investigated in further animal models.

GW24-e2512 A study on plasma PAI-1 and promoter-844 G- A,-675 4G/SG polymorphism of CHD

ObjectivesTo investigate the concentration of plasma plasminogen activator inhibitor-1 and the promoter-844G-A substitution,-675 4G/5G polymorphism in patients with CHD and control.Methods293 patients with CHD (UAP group andAMIgroup)and 173 human in...

Short-term effects of low-dose estrogen/drospirenone vs low-dose estrogen/dydrogesterone on glycemic fluctuations in postmenopausal women with metabolic syndrome

This study aims to compare the effects of low-dose emidrate estradiol/drospirenone (E2/DRSP) vs low-dose emidrate estradiol/dydrogesterone (E2/DG) combination on the mean amplitude of glycemic excursions (MAGE) value in postmenopausal women affected by metabolic syndrome (MS). One hundred sixty postmenopausal women were recruited to receive a treatment with oral doses of E2/1 mg plus drospirenone/2 mg (E2/DRSP group) or oral dose of E2/1 mg plus dydrogesterone/5 mg (E2/DG group) for 6 months. At enrollment and after 6 months, anthropometric, metabolic, and inflammatory parameters have been assessed. MAGE, evaluated during 48-h continuous subcutaneous glucose monitoring (CSGM), allowed us to assess daily glucose fluctuations at baseline and after 6 months. After hormone therapy, both groups showed a significant decline in fasting plasma glucose levels (p < 0.05), while only E2/DRSP group showed a statistically significant decline in waist circumferences, post-prandial glycemia, LDL, plasma triglycerides, MAGE, HOMA index, and plasma IL-6 (p < 0.05) levels. In the whole population (n = 160), after 6 months of indicated therapy, changes in fasting plasma glucose and PAI-1 levels correlated with the changes in MAGE values, while only in E2/DRSP group that MAGE reduction was positively associated with a stronger decrease in waist circumferences, triglycerides, and TNF-α plasma levels. The independent effect of hormone therapy (HT) on reduction in MAGE value has been tested in three different multiple linear regression models. HT resulted to be associated with MAGE, independent of other confounding variables. Although both groups had a decline in fasting plasma glucose, only drospirenone treatment revealed positive effects on glycemic excursions and insulin sensitivity, induced favorable changes in lipid profile, and showed an improvement of inflammatory indices in postmenopausal women with MS.

Abstract 150: Carotid Artery Atherosclerosis: Biomarkers of Plaque Instability

Objective To identify intraplaque and plasma biomarkers of carotid atherosclerotic plaque instability. We hypothesized that comparison of plaques and plasma from symptomatic and asymptomatic patients would show differences in proteolytic and inflammatory proteins, identifying them as potential markers of an unstable plaque. Methods 42 symptomatic and asymptomatic patients (22 women, 20 men, 52-89 years) presenting for carotid endarterectomy had blood samples drawn and carotid plaques processed for protein extraction. The intraplaque expression of proteolytic enzymes (uPA, tPA, PAI-1 and MMP-2) and inflammatory markers (TF, TSP-1, and TNF-α) were evaluated by Western blot analysis. Plasma analyte concentrations were assayed with LUMINEX CVD1 and Cytokine panels using LUMINEX technology. Results Intraplaque expression of tPA and uPA was 5.9x and 4.5x higher (n=15, p&lt; .05) respectively in the symptomatic group when compared to the asymptomatic group. The symptomatic group also demonstrated a 4.3x increase in active MMP-2 and a 3.6x increase in PAI-1 expression. Investigation of inflammatory proteins demonstrated 2.7-, 4.7-, and 3.3-fold increases in the expression of TNF- α, TSP-1, and TF, respectively. Plasma levels of MPO, MMP-9, IL8, MCP-1 and TNFα were significantly elevated in both the symptomatic and asymptomatic groups when compared to healthy controls (n=7, p&lt;0.05). While there was no change in ICAM-1 levels between asymptomatic patients and healthy controls, symptomatic patients demonstrated a 35% decrease (n=7, p&lt;0.05) when compared to controls. Also, plasma PAI-1 levels in asymptomatic patients were 55% lower than in symptomatic patients and 75% lower than in healthy controls. Conclusion Elevated expression of certain inflammatory markers and proteolytic enzymes in carotid plaques was demonstrated in symptomatic patients. Marked differences were also observed in ICAM-1 and PAI-1 expression in the plasma of symptomatic patients relative to asymptomatic patients and healthy controls. This leaves promising avenues for further inquiry into the roles of these proteins in atherosclerotic disease and their potential as clinical biomarkers of an unstable atherosclerotic plaque.

TPA Alu (I/D) Polymorphism Associates With Bacterial Osteomyelitis

Coagulation and fibrinolysis are important in infections and systemic inflammatory response syndrome. Polymorphisms in plasminogen activator inhibitor-1 (PAI-1, SERPINE1) and tissue plasminogen activator (tPA, PLAT), such as PAI-1 (-675 4G/5G deletion/insertion) and tPA (Alu insertion/deletion [I/D]), are associated with strokes, myocardial infarctions, bacterial infections and septic shock severity, and trauma. Osteomyelitis is a mostly posttraumatic, Staphylococcal bone infection. tPA Alu (I/D) (rs4646972) and PAI-1 (4G/5G) (rs1799889) polymorphisms were studied by DNA amplification with polymerase chain reaction in 261 patients with osteomyelitis and in 299 matched blood donors. Plasma PAI-1/tPA complex was assessed by enzyme-linked immuosorbent assay. II homozygotes (37.9% vs 19.1%) and I allele carriers (56.3% vs 46.3%) for the tPA Alu (I/D) polymorphism were significantly more frequent in osteomyelitis patients compared to controls (P < .001). II genotype carrier osteomyelitis patients had lower PAI-1/tPA complex levels compared to those with the D allele (P ≤ .04). There was no association between these genotypes and chronicity of osteomyelitis, post-traumatic etiology, or with a specific bacterial etiology. PAI-1 (4G/4G) homozygotes were not significantly different between osteomyelitis patients and controls (P = .1). We report for the first time to our knowledge an association between the tPA Alu (I/D) polymorphism and susceptibility to bacterial osteomyelitis, perhaps by fibrinolysis dysfunction.

Plasminogen activator inhibitor-1 in children with central obesity: Effect on left ventricular function

Aim The aim of the study was to investigate the Plasminogen activator inhibitor-1 (PAI-1) expression in obese children and to clarify its role with respect to left ventricular (LV) function. Patients and methods This study included 69 obese children and adolescents, 40 lean healthy controls. Children were considered obese according to body mass index (BMI) percentile for age and sex curves of growth for our population. Each subject’s fat distribution was assessed by measuring waist hip ratio (WHR). Obese children with peripheral fat distribution were excluded. Exclusion criteria included hypertension, endocrine, cardiovascular, renal, insulin dependent or independent diabetes mellitus and smoking habits. Laboratory investigations included measurement of plasma PAI-1 antigen. Determination of total serum cholesterol, low density lipoprotein, high density lipoprotein, triglycerides, blood glucose and fasting serum insulin. Echocardiography study was obtained by two dimensionally guided M mode. Results BMI and WHR were significantly higher in obese compared to lean children (P 0.05). Plasma PAI-1 were significantly higher in obese compared to controls (P = 0.03). A significant direct correlation was revealed between PAI-1 in comparison to WHR, fasting insulin and LVM/H. Plasma PAI -1 and WHR were independent predictors of LVM/H. Conclusions Obese children with central fat distribution showed an increase in plasma PAI-1 antigen. Also PAI-1 contributes directly to the complication of obesity including type 2 diabetes and cardiovascular disease.

The role of plasminogen activator inhibitor-1 in gastric mucosal protection

Gastric mucosal health is maintained in response to potentially damaging luminal factors. Aspirin and nonsteroidal anti-inflammatory drugs (NSAIDs) disrupt protective mechanisms leading to bleeding and ulceration. The plasminogen activator system has been implicated in fibrinolysis following gastric ulceration, and an inhibitor of this system, plasminogen activator inhibitor (PAI)-1, is expressed in gastric epithelial cells. In Helicobacter pylori-negative patients with normal gastric histology taking aspirin or NSAIDs, we found elevated gastric PAI-1 mRNA abundance compared with controls; the increase in patients on aspirin was independent of whether they were also taking proton pump inhibitors. In the same patients, aspirin tended to lower urokinase plasminogen activator mRNA. Immunohistochemistry indicated PAI-1 localization to epithelial cells. In a model system using MKN45 or AGS-GR cells transfected with a PAI-1 promoter-luciferase reporter construct, we found no evidence for upregulation of PAI-1 expression by indomethacin, and, in fact, cyclooxygenase products such as PGE2 and PGI2 weakly stimulated expression. Increased gastric PAI-1 mRNA was also found in mice following gavage with ethanol or indomethacin, but plasma PAI-1 was unaffected. In PAI-1−/− mice, gastric hemorrhagic lesions in response to ethanol or indomethacin were increased compared with C57BL/6 mice. In contrast, in PAI-1-H/Kβ mice in which PAI-1 is overexpressed in parietal cells, there were decreased lesions in response to ethanol and indomethacin. Thus, PAI-1 expression is increased in gastric epithelial cells in response to mucosal irritants such as aspirin and NSAIDs probably via an indirect mechanism, and PAI-1 acts as a local autoregulator to minimize mucosal damage.