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PubMed2004

Biomaterial-associated thrombosis: roles of coagulation factors, complement, platelets and leukocytes.

Our failure to produce truly non-thrombogenic materials may reflect a failure to fully understand the mechanisms of biomaterial-associated thrombosis. The community has focused on minimizing coagulation or minimizing platelet adhesion and activation. We have infrequently considered the interactions between the two although we are generally familiar with these interactions. However, we have rarely considered in the context of biomaterial-associated thrombosis the other major players in blood: complement and leukocytes. Biomaterials are known agonists of complement and leukocyte activation, but this is frequently studied only in the context of inflammation. For us, thrombosis is a special case of inflammation. Here we summarize current perspectives on all four of these components in thrombosis and with biomaterials and cardiovascular devices. We also briefly highlight a few features of biomaterial-associated thrombosis that are not often considered in the biomaterials literature: The importance of tissue factor and the extrinsic coagulation system. Complement activation as a prelude to platelet activation and its role in thrombosis. The role of leukocytes in thrombin formation. The differing time scales of these contributions.

Biocompatible Materials↗
PubMed2005

Thrombectomy in HIV related peripheral arterial thrombosis: a preliminary report.

OBJECTIVES: To evaluate the limb salvage rate in patients undergoing thrombectomy for HIV related peripheral arterial thrombosis. METHODS: A prospective review of patients with HIV related peripheral arterial thrombosis managed at the Durban Metropolitan Vascular Service in Kwazulu-Natal South Africa over a 5-year (1998-2003) period. All patients underwent thrombectomy. RESULTS: Twenty-two patients (20 males) with HIV related peripheral arterial thrombosis (two upper limbs, 20 lower limbs) were included in the study. The median age was 36 years (24-46 years). All patients had rest pain and seven patients had gangrene (digital five, forefoot two). Seven patients had a short history of claudication before development of rest pain. Mean duration of symptoms was 30 days (1-120 days) with 10 patients presenting within 24 h. Most patients had a good general state of health and only three had clinical evidence of HIV infection. Ten patients who presented with threatened limbs from acute arterial occlusion were subjected to thrombectomy without any diagnostic investigations. Twelve patients who did not present with critical limb ischaemia had duplex ultrasonography, which showed arterial occlusion by a thrombus with normal proximal arteries. The striking features were the normal proximal vessels and absence of distal run-off. The search for an underlying cause, echocardiography in seven patients and coagulation screening in 10 patients, was always negative. All patients were subjected to thrombectomy with an on-table angiogram and received systemic heparin intra-operatively and deltaparin post-operatively. In 20 patients, duplex ultrasonography confirmed re-thrombosis within 48 h. Two patients remained with patent arteries. Most (16/20) patients who re-thrombosed required a major amputation and thrombectomy did not alter the level of amputation. The other four patients with re-thrombosis had symptomatic relief. Three patients died within 30 days of thrombectomy, all of HIV related complications. Overall the limb salvage rate was 6/22 (27%). CONCLUSION: Limb salvage rate following thrombectomy in HIV related peripheral arterial thrombosis is very low. Duplex ultrasonography appears to be an adequate radiological diagnostic investigation for these patients.

Adult↗
PubMedDate not supplied

Right portal vein thrombosis after splenectomy for trauma.

Portal vein thrombosis may complicate splenectomy in patients with hemolytic anemia and myeloproliferative disease, whereas the frequency of portal vein thrombosis in case of trauma is not defined. A case of right portal vein thrombosis after splenectomy for trauma is reported in this paper. Hematologic workup did not reveal an underlying platelet or coagulation disorder. The patient was promptly anti-coagulated with complete recanalization of the portal vein. We conclude that mild symptoms, like abdominal pain and fever, after splenectomy should be investigated with a color Doppler ultrasonography to confirm or rule out a diagnosis of portal thrombosis and to anti-coagulate the patient with thrombosis, thus preventing bowel infarction and secondary portal hypertension. Routine postoperative color Doppler might also be justified in all postsplenectomy patients (without hematologic diseases) for early detection of a portal vein thrombosis.

Abdominal Injuries↗
PubMed2004

Fetal thrombotic vasculopathy in the placenta: a thrombophilic connection between pregnancy complications and neonatal thrombosis?

OBJECTIVE: Fetal thrombotic vasculopathy (FTV) has been related to pregnancy complications and neonatal thrombosis separately. We assessed whether a relationship existed in our population of women with neonates who were admitted to our Neonatal Intensive Care Unit (NICU). In addition, the presence of thrombophilic factors in children and parents was investigated. METHODS: Two groups were detected by a search of the departmental databases. Group A was a cohort of 5000 neonates admitted to our NICU (1992-2002). Infants who developed thrombotic complications were selected. Group B was a cohort of placentae from our institution (2000, n = 141). Those with a diagnosis of FTV were selected. Case-notes and laboratory results were obtained through the hospital information system. RESULTS: Of Group A, thrombosis was reported in 55 children. Of these, 20 matching placentae were available. Eight placentae showed FTV (40 per cent). Of the eight corresponding pregnancies, seven were complicated by pre-eclampsia and/or intra uterine growth restriction (IUGR). Of the 12 placentae without FTV, five of the pregnancies had pre-eclampsia and/or IUGR (odds ratio for relation FTV-Complications: 9.8, 95 per cent CI = 0.9-107). In Group B, nine placentae showed FTV (6.4 per cent). Of these nine, six of the pregnancies were complicated by pre-eclampsia and/or IUGR. None of the neonates developed thrombosis. CONCLUSION: Pre-eclampsia and/or IUGR as well as neonatal thrombosis are both associated with fetal thrombotic vasculopathy in the placenta. However, in our selected-tertiary centre-population, FTV did not predict neonatal thrombosis. The thrombophilic investigations of parents and children were incomplete. A standard approach for evaluating parents at risk for FTV and evaluating neonates at risk for thrombosis should be developed.

Adult↗
PubMed2005

Umbilical arterial thrombosis with vascular wall necrosis: clinicopathologic findings of 11 cases.

Thrombosis of umbilical cord vessels is associated with a high perinatal mortality. Most cases are venous thrombi, arterial thrombi are uncommon. To clarify clinical and pathological features of this entity, we reviewed 11 cases with umbilical artery thrombosis. The gestational ages were 33-40 (mean 36.8) weeks. Three cases (38%) were associated with severe intrauterine growth retardation and two cases (25%) led to intrauterine fetal deaths. All cases had completely occlusive thrombi of one umbilical artery. The arteries with thrombosis demonstrated partial necrosis of vascular wall. The pathogenesis is unclear, but nine cases (82%) had cord abnormalities (long cord, peripheral cord insertion, short cord with twist, funisitis). This paper is the first report of clinical and pathological features of umbilical artery thrombosis. The results of this study confirm the clinical importance and unique histological findings of umbilical artery thrombosis. The recognition of umbilical artery thrombosis is necessary to establish the diagnosis and treatment of this condition.

Adult↗
PubMed2003

Fish oil inhibits photochemically induced thrombosis in the guinea pig in a dose dependent manner.

The dose-response effect of dietary fish oil was investigated in the photochemically induced thrombosis model in guinea pigs. In this arterial thrombosis model thrombus formation was evaluated by determination of different occlusion parameters (percentage of occlusion, area under the blood flow curve, time to first occlusion, spontaneous reflow). Sixty guinea pigs (7 weeks old) were randomly assigned to and fed a 40 energy % diet containing increasing amounts (0, 5.5, 17 and 36 energy %) of fish oil for four weeks. Arterial thrombosis was induced in the femoral artery by free radical damage and subsequent thrombus formation. Increasing fish oil concentrations in the diet were associated with a linear decrease (p<0.001) in the percentage of occlusion (calculated as a decrease in blood flow) and a linear increase in area under the blood flow curve/begin flow (p<0.001). The time to thrombus formation was not significantly prolonged in any group. However the frequency of animals in which complete occlusion of the femoral artery was not obtained during the thrombosis induction and subsequent observation period was higher in the groups receiving the two highest doses of fish oil. Spontaneous reflow correlated positively (p<0.013) with increasing dietary fish oil content. In conclusion, our data indicates that dietary fish oil inhibits photochemically induced thrombosis in this animal model of arterial thrombosis in a dose dependent manner.

Animal Feed↗
PubMed2006

An unfavorable combination of Factor V Leiden with age, weight, and blood group causes high risk of pregnancy-associated venous thrombosis: a population-based nested case-control study.

INTRODUCTION: Hereditary and acquired risk factors increase the risk for thrombosis among pregnant women. Few risk estimates are, however, well established. The aim of the present study was to assess risk for pregnancy-associated venous thrombosis of factor V Leiden (FVL), FII G20210A, FV A4070G, MTHFR C677T, TFPI C536T, PROC T38853G, FXIII V34L, blood group, age, and body mass index (BMI), and their interactions and public health impact. MATERIALS AND METHODS: Study design is a population-based nested case-control study of 100,000 consecutive pregnancies in Finland. Cases and controls were identified by combining national registers. Thirty four cases with objectively diagnosed venous thrombosis and 641 controls were studied. RESULTS: FVL (OR 11.6, 95% CI 3.6-33.6), age >35 vs. <25 (OR 6.3, 95% CI 1.7-23.1), and BMI >30 vs. <25 (OR 5.6, 95% CI 2.3-13.9) were associated with thrombosis. Overall absolute risk of a FVL carrier was 1 in 314. FVL interacted with age, BMI, and blood group. Population attributable risk proportion was 19% for FVL, 23% for age >35, 33% for BMI >25, and 35% for non-O blood group. Unexpectedly, the prevalence of FVL increased with age in controls. CONCLUSIONS: FVL appeared as a strong risk factor for pregnancy-associated venous thrombosis. Especially in elderly overweight mothers, FVL may cause a substantial thrombosis risk. Further studies are needed to confirm the increased prevalence of FVL in elderly mothers with normal pregnancies.

Adolescent↗
PubMed2006

The prevalence of antibodies to the platelet factor 4 -heparin complex and association with access thrombosis in patients on chronic hemodialysis.

INTRODUCTION: Heparin-induced thrombocytopenia is a serious complication that can lead to thrombocytopenia, venous and arterial thrombosis. Patients with this disorder develop antibodies to the platelet factor 4-heparin (PF4-H) complex. Hemodialysis patients are repeatedly exposed to heparin and are at risk for developing PF4-H antibodies. We sought to determine the prevalence of PF4-H antibodies in a large cohort of patients on chronic hemodialysis and to evaluate the relationship between PF4-H antibodies and hemodialysis vascular access thrombosis in a case-control study. MATERIAL AND METHODS: Pre-dialysis blood samples were drawn on 419 patients; 107 cases with access thrombosis and 312 controls that never had access thrombosis. All samples were screened for PF4-H antibodies using an ELISA assay (GTI PF4 Enhanced, GTI Diagnostics). All positive and indeterminate samples were then tested using an IgG-specific PF4-H ELISA assay and a platelet serotonin-release assay. RESULTS: Antibodies to PF4-H were positive in 54 (12.9%) patients using the screening ELISA assay. Nine (2.1%) patients had IgG-specific PF4-H antibodies. None of the patient's had a positive platelet serotonin-release assay. No relationship between hemodialysis access thrombosis and PF4-H antibodies was noted using the screening ELISA assay (unadjusted odds ratio 0.63; 95% CI 0.30-1.30; P = 0.21), the IgG-specific ELISA assay (unadjusted odds ratio 0.83; 95% CI 0.17-4.06; P = 0.82) or indeterminate platelet serotonin-release assay results (unadjusted odds ratio 0.97;95% CI 0.10-9.44;P = 0.98). CONCLUSIONS: Hemodialysis with repeated exposure to unfractionated heparin was associated with a moderately elevated prevalence of PF4-H antibodies. However, our results do not support a relationship between PF4-H antibodies and hemodialysis vascular access thrombosis.

Adult↗
PubMed2001

Comparison of four strategies for diagnosing deep vein thrombosis: a cost-effectiveness analysis.

PURPOSE: Four strategies for the diagnosis of deep vein thrombosis have been validated recently. The strategies use various combinations of assessment of a patient's clinical probability of having deep venous thrombosis, serial lower limb venous compression ultrasonography, and measurement of plasma D-dimer levels. We compared the cost-effectiveness of these diagnostic strategies. MATERIALS AND METHODS: We performed a formal cost-effectiveness analysis using a decision-analysis model. Outcomes considered were costs per patient, 3-month quality-adjusted survival, number of lives saved per 1,000 patients, and incremental costs per quality-adjusted life-year (QALY) gained. RESULTS: Under baseline conditions, with a 24% prevalence of deep vein thrombosis in tested patients, the effectiveness of all strategies was similar (4.6 to 4.8 lives saved per 1,000 patients managed). The most expensive strategy was serial ultrasound (repeat ultrasound on day 7 in all patients with a normal initial ultrasound) at a cost-effectiveness of $10,716 per additional QALY. Performing a repeat ultrasound only in patients with an elevated D-dimer level (serial ultrasound with D-dimer) was somewhat less expensive at $10,281 per additional QALY. Taking clinical probability into account by repeating ultrasound only in patients with an intermediate clinical probability of deep vein thrombosis (risk-based serial ultrasound) yielded further savings and cost $10,090 per additional QALY. The least expensive and most cost-effective option was to perform D-dimer as the initial test, followed by a single ultrasound if the D-dimer level was abnormal, and by phlebography in patients with a normal ultrasound and a high clinical probability of deep vein thrombosis (D-dimer with risk-based single ultrasound) at $8,897 per additional QALY. This strategy allowed a 17% reduction in incremental costs compared with the most expensive algorithm and reduced resource consumption (70 ultrasound procedures per 100 patients managed vs 130 to 170 with the other diagnostic strategies). CONCLUSIONS: Combining clinical probability and D-dimer with a single ultrasound in the diagnostic workup of patients with possible deep vein thrombosis is highly cost-effective, allowing a reduction in costs and resource use without any substantial increase in mortality. Serial ultrasonography is less cost-effective.

Algorithms↗
PubMed2002

Usefulness of a semiquantitative D-dimer test for the exclusion of deep venous thrombosis in outpatients.

PURPOSE: The D-dimer test is used commonly in diagnostic strategies to reduce the need for ultrasonography in patients suspected of having deep venous thrombosis. We studied several clinical and laboratory variables that might limit the accuracy of a semiquantitative D-dimer test. SUBJECTS AND METHODS: In this retrospective cohort study, 704 outpatients suspected of having deep venous thrombosis underwent a semiquantitative D-dimer test and ultrasonography. The performance of the D-dimer test was calculated in patients using anticoagulants (n =61), patients with previous thrombosis (n =127), and patients with malignancy (n =47), including 39 patients with more than one of these characteristics. The 508 remaining patients were considered to be the reference group. RESULTS: A total of 254 patients (36%) had evidence of deep venous thrombosis. The D-dimer test had a sensitivity of 99% (174/176; 95% confidence interval [CI]: 96% to 100%) and a negative predictive value of 98% (98/100; 95% CI: 93% to 100%) in the reference group. The sensitivity of the D-dimer test in patients using oral anticoagulants was 75% (6/8; 95% CI: 35% to 97%; P =0.01 compared with the reference group). Test sensitivity was 96% (51/53; 95% CI: 87% to 100%) in patients with previous thrombosis, and 100% (29/29; 95% CI: 88% to 100%) in patients with cancer. However, 553 (79%) of all patients, including 43 of the cancer patients (91%), had an abnormal D-dimer test. CONCLUSION: The semiquantitative D-dimer test in this study has a high sensitivity and negative predictive value in the exclusion of deep venous thrombosis, except perhaps among patients using oral anticoagulants. D-dimer tests in patients with cancer and in patients over 70 years old may not be worthwhile, because the tests are usually positive.

Age Distribution↗
PubMed2002

Practical diagnostic management of patients with clinically suspected deep vein thrombosis by clinical probability test, compression ultrasonography, and D-dimer test.

PURPOSE: To evaluate a new noninvasive diagnostic strategy for ruling out deep vein thrombosis consisting of either a combination of low clinical probability and normal ultrasonography or a combination of moderate-to-high clinical probability, normal ultrasonography, and a normal D-dimer test. SUBJECTS AND METHODS: We studied 811 patients with clinically suspected deep vein thrombosis using a diagnostic management strategy that combined clinical probability, ultrasonography, and measurement of D-dimers. The primary endpoint was venous thromboembolism occurring during a 3-month follow-up. RESULTS: Of the 280 patients (35%) with a low clinical probability, 30 (11%) had an abnormal initial ultrasonography and were treated. Of the other 250 untreated patients with low clinical probability and a normal ultrasonography, 5 (2%; 95% confidence interval [CI]: 1% to 5%) developed a nonfatal venous thromboembolism during follow-up. Of the 531 patients (65%) with a moderate-to-high clinical probability, 300 (56%) had an abnormal ultrasonography. Of the remaining 231 patients with a normal ultrasonography, 148 had a normal D-dimer test; none of these patients developed deep vein thrombosis during follow-up (0%; 95% CI: 0% to 3%). Of the 83 patients with an abnormal D-dimer test, 77 underwent repeat ultrasonography about 1 week later; none of the 64 patients with a second normal ultrasound developed symptomatic deep vein thrombosis during follow-up (0%; 95% CI: 0% to 6%). CONCLUSIONS: This management strategy, which combines clinical probability, ultrasonography, and D-dimer measurements, is practical and safe in ruling out deep vein thrombosis in patients with clinically suspected thrombosis and reduces the need for repeat ultrasonography.

Adult↗
PubMed1992

Pregnancy loss and thrombosis with protein C deficiency.

OBJECTIVES: Protein C inhibits coagulation and promotes fibrinolysis. This study investigates the association between protein C deficiency and pregnancy loss, thrombosis in pregnancy, and thrombosis with oral contraception. STUDY DESIGN: Fifteen protein C--deficient patients and 37 controls from a single kindred were studied. An obstetric history was obtained by telephone. Data were analyzed by logistic regression, Fisher's exact test, and Student t test. RESULTS: Protein C--deficient women experienced a 33% pregnancy loss versus 19% in the controls (not significant). Thromboembolism during pregnancy in protein C--deficient women was 33% (45% in those not receiving prophylactic anticoagulation) versus 5% in controls (odds ratio 7.37, p = 0.026). Five of 12 protein C--deficient women using oral contraception developed thrombosis versus 0 of 33 controls. The risk of thrombosis for protein C--deficient women using oral contraception is increased (p < 0.001). CONCLUSIONS: Perinatal outcome is not statistically different with protein C deficiency. Protein C deficiency increases the risk of thrombosis during pregnancy and with oral contraception. Prophylactic heparin is suggested during pregnancy for protein C--deficient women with personal or family histories of thrombosis. Oral contraception is not advised.

Adult↗
PubMed1999

The preoperative prevalence of deep vein thrombosis in patients with femoral neck fractures and delayed operation.

Out of 61 consecutive patients admitted for femoral neck fracture 21 patients had a delay to operation for more than 48 h from the time of injury. We studied these patients prospectively for the presence of deep-vein thrombosis (DVT). 13 (62%) had venographic evidence of thrombosis. All occurred in the broken limb. Five patients had bilateral thrombosis. The delay alone seems to be the major risk factor for thrombosis irrespective of age, fracture type, premorbid mobility and coexisting illness. The prevalence of preoperative DVT 48 h after injury approaches the reported postoperative incidence of DVT, which suggests that DVT will occur in a high proportion of patients regardless of treatment and prophylaxis. We recommend that those patients, in whom operation is delayed, should be routinely investigated for the presence of thrombosis preoperatively and a prophylactic vena cava filter should be considered when major deep vein thrombosis occurred.

Aged↗
PubMed2002

Pediatric thrombosis.

Although thrombosis is less frequent in children than in adults, it represents a significant source of morbidity and mortality. Multiple factors. both genetic and acquired. contribute to the development of thrombosis in chiidren. Thrombosis in a child warrants investigation of potential underlying prothrombotic conditions. The risk of thrombosis in children with heterozygous deficiencies is not clearly defined, but it appears that children who are heterozygous for more than one risk factor or who have a combination of inherited and acquired defects are at higher risk for thrombosis. Treatment of thrombosis primarily involves a rapidly acting anticoaguiant such as heparin or LMWH to prevent extension, and long-term anticoagulation with warfarin may be instituted to prevent recurrence. Thrombolytic therapy with recombinant tissue plasminogen activator also appears to be safe and effective in children. Prospective and multicenter studies are still needed to clarify the contribution of specific prothrombotic disorders to childhood TE so that evidence-based treatment recommendations can be made.

Child↗
PubMed2003

Size effect of intratracheally instilled particles on pulmonary inflammation and vascular thrombosis.

Particulate air pollution is associated with cardiorespiratory effects and ultrafine particles (UFPs, diameter < 100 nm) are believed to play an important role. We studied the acute (1 h) effect of intratracheally instilled unmodified (60 nm), negatively charged carboxylate-modified (60 nm), or positively charged amine-modified (60 or 400 nm) polystyrene particles on bronchoalveolar lavage (BAL) indices and on peripheral thrombosis in hamster. The latter was assessed by measuring the extent of photochemically induced thrombosis in a femoral vein via transillumination. Unmodified and negative UFPs did not modify thrombosis and BAL indices. Positive UFPs increased thrombosis at 500 microg per animal (+ 341 +/- 96%) and at 50 microg per animal (+ 533 +/- 122%), but not at 5 microg per animal. Neutrophils, lactate dehydrogenase, and histamine were increased in BAL at all these doses but protein concentration was increased only at 500 microg per animal. Positive 400-nm particles (500 microg per animal) did not affect thrombosis, although they led to a neutrophil influx and an increase in BAL proteins and histamine. Using the Platelet Function Analyser (PFA-100), the platelets of hamsters were activated by the in vitro addition of positive UFPs and 400-nm particles to blood. We conclude that intratracheally administered positive ultrafine and 400-nm particles induce pulmonary inflammation within 1 h. Positive UFPs, but not the 400-nm particles enhance thrombosis. Hence, particle-induced lung inflammation and thrombogenesis can be partially uncoupled.

Air Pollutants↗
PubMed2000

Microvascular consequences of thrombosis in small venules: an in vivo microscopic study using a novel model in the ear of the hairless mouse.

Little is known of the development of chronic microvascular alterations following small vessel thrombosis, which is probably due to the lack of appropriate experimental models. Herein we report the first results on thrombosis-associated long-term changes of microvascular permeability and vessel tortuosity and diameter and blood perfusion using the ear of the hairless mouse, and intravital fluorescence microscopy. Thrombosis was induced photochemically in small venules (diameter: 75 to 100 microm) using light/dye exposure (fluorescein isothiocyanate-dextran 150,000), and the microcirculation compromised by the blockade of blood drainage was analyzed before and 30 minutes after induction of thrombosis as well as repeatedly over a 28-day observation period. Thrombus formation resulted in a marked increase (p<0.05) of microvascular permeability (0.85+/-0.11) when compared with baseline values (0.46+/-0.04). Permeability remained elevated (p<0.05) at days 1 (0.67+/-0.07), 3 (0.58+/-0.02), and 7 (0.60+/-0.06), but returned to normal after 28 days (0.43+/-0.03). Tortuosity, diameter, and red blood cell velocity of venular segments, located upstream of thrombus formation, were found unchanged during the entire 28-day observation period. This was probably due to the fact that blood flow from the thrombosis-affected tissue was frequently drained into nonaffected tissue via preexisting "through-fare" channels, serving as venulo-venular collaterals. In accordance, in 10 to 20% of these venular segments the direction of blood perfusion was found changed, while those changes were only rarely observed in venular vessel segments of the nonthrombotic contralateral ears. We conclude that thrombosis in small cutaneous venules is primarily characterized by an increased vascular permeability, reflecting an inflammatory response, similar to what is known from thrombophlebitis in patients. The model presented herein may be a versatile tool to study pathogenesis of chronic microcirculatory derangements in microthrombosis and their prevention by novel therapeutic strategies.

Animals↗
PubMed2001

A novel platelet-rich arterial thrombosis model in rabbits. Simple, reproducible, and dynamic real-time measurement by using double-opposing inverted-sutures model.

Though numerous animal thrombosis models have been introduced, an easy, reliable, and reproducible arterial thrombosis model remains a continuing challenge prior to a thrombolytic study. In an effort to evaluate the efficiency of various recombinant thrombolytic agents with specific affinity to activated platelets in vivo, we developed a novel double-opposing inverted-sutures model to create a platelet-rich thrombus in the femoral artery of rabbits. The arteriotomy was done semicircumferentially, and variously sized microsurgical sutures were introduced intraluminally in a double-opposing inverted manner. The animals were divided into three groups according to the double-opposing inverted-sutures used: Group 1 with 10-0 nylon (n=6), Group 2 with 9-0 nylon (n=6), and Group 3 with 8-0 nylon (n=22). The superficial epigastric branch was cannulated with a thin polyethylene (PE) tube for intraarterial administration of the studied thrombolytic agent. The blood flow was continuously measured with a real-time ultrasonic flow meter. Within 2 h of installation of the sutures, there was no thrombus formation in either Group 1 or 2. In Group 3, the thrombosis rate was 91% (20 of 22) under a steady baseline flow (with an average of 12.23+/-2.40 ml/min). It was highly statistically significant with a P-value of .0000743 using Fisher's Exact Test. The averaged time to thrombosis was 21.8+/-9.8 min. The ultrasonic flow meter to record the dynamic real-time measurement of blood flow was a guideline for thrombus formation or dissolution, which was correlated with the morphological findings of stenotic status of the vessel detected by the Doppler sonography. The components of the thrombus were proven to be platelet-rich predominant by histological examination via hematoxylin and eosin (H&E) stain and transmission electron microscopy (TEM). To confirm that the double-opposing inverted-sutures model would be useful for a study of thrombolytic agents, we evaluated the effects of recombinant tissue-type plasminogen activator (rt-PA) and streptokinase-human plasminogen (SK-HPlg). The average time to thrombolysis post rt-PA infusion was 16.0+/-8.2 min and that of SK-HPlg was 79.6+/-23.1, which were similar to the previous reports. In conclusion, the novel double-opposing inverted-sutures (8-0 nylon) model provides a simple, reliable, and reproducible platelet-rich arterial thrombosis model with noninvasive and dynamic real-time measurement. It may be applied in assessing the efficiency of the recombinant thrombolytic agents and offers many advantages of an arterial platelet-rich in vivo thrombosis model.

Animals↗
PubMed2001

Antithrombin deficiency in Brazilian patients with venous thrombosis: molecular characterization of a single splice site mutation, an insertion and a de novo point mutation.

The prevalence of antithrombin (AT) deficiency in 342 unselected Brazilian patients with venous thrombosis was 1.16%, which increased to 3% when only patients under the age of 50 or with a familial history of thrombosis were considered. In two patients, a clinical (contraceptive use) or genetic risk factor (factor V Leiden and C677T in the methylene tetrahydrofolate reductase gene [MTHFR]) was identified and corroborated the hypothesis that an interaction of factors accounted for the appearance of thrombosis. However, no risk factor other than AT deficiency was identified in one patient with an important clinical and family history of spontaneous thrombosis. Three mutations were identified in these patients: a G-->A transition in intron 5 at position +1 (5'-->3'), three base insertions corresponding to arginine at position 5383 in exon 3A, and a G-->A transition at 13328, corresponding to an Ala404Thr de novo mutation. The polymorphisms in the genes coding for coagulation factors XII and XIII and fibrinogen normally associated with an increased risk for venous thrombosis were not related to thrombosis in these patients. This is the first study in South America to assess the prevalence of AT deficiency and to report the molecular characterization of the mutations involved.

Adult↗