Heparin Fractions Research Articles

2.5 kDa and 5.0 kDa heparin fragments specifically inhibit microvessel sprouting and network formation in VEGF165-mediated mammalian angiogenesis.

Tumour growth is angiogenesis dependent. Thrombosis and thromboembolism are very common in cancer patients. These patients are often treated with heparin as an anti-coagulant. Many tumour angiogens, including VEGF165, and endogenous anti-angiogenesis factors bind heparin tightly. Using the non-surgical mesenteric-window angiogenesis assay, we studied in detail the systemic effect of heparin fractions with a mean MW of 2.5, 5.0 and 16.4 kDa on the microvessel sprouting and network formation in angiogenesis mediated by VEGF165 in rats. The microvessel network was assessed objectively in terms of the number and lengths of segments (the distance between two successive branching points), the number of branching points, the degree of tortuosity, the index of interconnecting loop formation, the index of intersection, as well as the number and lengths of sprouts. Compared with the saline control, the 2.5 kDa fraction significantly shortened the microvessel sprouts and the microvessel segments but increased the microvessel tortuosity in statistical terms; the 5.0 kDa fraction statistically significantly shortened the sprouts, decreased the number of segments and the number of microvessel branching points; whereas the 16.4 kDa fraction statistically significantly elongated the longest segments. Moreover, statistically significant differences were found between the three heparin fractions in terms of microvessel tortuosity (2.5 vs. 16.4 kDa), index of loop formation (5.0 vs. 2.5 + 16.4 kDa) and index of intersection (5.0 vs. 16.4 kDa). These findings demonstrate that heparin fragments size-specifically inhibit microvessel sprouting and network formation in VEGF165-mediated angiogenesis. As VEGF165 is a potent angiogen in human tumours, we suggest that heparin enriched in 2.5 kDa species and 5.0 kDa species especially should be exploited as a combined anti-coagulant and specific adjuvant anti-angiogenic agent in cancer patients who require anti-coagulant therapy.

Does Perioperative Antithrombotic Therapy Increase the Likelihood of a Postoperative Coagulopathy After Cardiac Surgery?

Washington University School of MedicineAssistant Professor of AnethesiologyDepartment of Anesthesiologyskubasn@notes.wustl.eduAssociate Professorof Anesthesiology and PathologyWashington University School of MedicineSt. Louis, Missouri 63110In Reply:—We agree with Drs. Lewis and Collard that measurement of anti-Xa activity and platelet function would have provided important information. Nevertheless, in the presence of normal ACT, no detectable heparin by heparin concentration assay (Hepcon, Medtronic Blood Management, Parker, CO), no identifiable surgical source, and a borderline platelet count, possible explanations for this patients’ excessive and protracted chest tube output that was unresponsive to hemostatic blood product transfusion, include either the persistent anticoagulant properties of low molecular weight heparin i.e., anti-Xa activity, or a platelet function abnormality that may be related to either of these agents, or a possible interaction between low molecular weight heparins and tirofiban. Previous anecdotal reports of increased bleeding when patients had received either low molecular weight heparin preparations 1,2or platelet inhibitors 3–5support our findings.Furthermore, we are uncertain of the potential beneficial effects of hemofiltration, because we have previously shown that hemofiltration (using one specific filter) during cardiopulmonary bypass failed to remove the lower molecular weight fraction of unfractionated heparin (no anti-Xa activity in ultrafiltrate). 6Factors related to the low molecular weight heparin molecule, such as, its linear shape, electrostatic charge, or binding to antithrombin III may account for these findings. Therefore, we would recommend that in patients receiving either enoxaparin or tirofiban, postponement of elective cardiac surgery should be considered for a period of time well over the five half-lives of either medication. Another alternative would be to switch patients from low molecular weight heparin to unfractionated heparin as soon as an operation is proposed. Further studies are needed to evaluate the efficacy of hemofiltration or plasma exchange filters with respect to removal of these agents.

Inhibition of allergic late airway responses by inhaled heparin-derived oligosaccharides.

Inhaled heparin has been shown to inhibit allergic bronchoconstriction in sheep that develop only acute responses to antigen (acute responders) but was ineffective in sheep that develop both acute and late airway responses (LAR) (dual responders). Because the antiallergic activity of heparin is molecular-weight dependent, we hypothesized that heparin-derived oligosaccharides (<2, 500) with potential anti-inflammatory activity may attenuate the LAR in the dual-responder sheep. Specific lung resistance was measured in 24 dual-responder sheep before and serially for 8 h after challenge with Ascaris suum antigen for demonstration of early airway response (EAR) and LAR, without and after treatment with inhaled medium-, low-, and ultralow-molecular-weight (ULMW) heparins and "non-anticoagulant" fractions (NAF) of heparin. Airway responsiveness was estimated before and 24 h postantigen as the cumulative provocating dose of carbachol that increased specific lung resistance by 400%. Only ULMW heparins caused a dose-dependent inhibition of antigen-induced EAR and LAR and postantigen airway hyperresponsiveness (AHR), whereas low- and medium-molecular-weight heparins were ineffective. The effects of ULMW heparin and ULMW NAF-heparin were comparable and inhibited the LAR and AHR even when administered "after" the antigen challenge. The ULMW NAF-heparin failed to inhibit the bronchoconstrictor response to histamine, carbachol, and leukotriene D(4), excluding a direct effect on airway smooth muscle. In six sheep, segmental antigen challenge caused a marked increase in bronchoalveolar lavage histamine, which was not prevented by inhaled ULMW NAF-heparin. The results of this study in the dual-responder sheep demonstrate that 1) the antiallergic activity of inhaled "fractionated" heparins is molecular-weight dependent, 2) only ULMW heparins inhibit the antigen-induced EAR and LAR and postantigen AHR, and 3) the antiallergic activity is mediated by nonanticoagulant fractions and resides in the ULMW chains of <2,500.

Development of Tinzaparin: A Heparinase-Digested Low-Molecular-Weight Heparin

Although heparin and its properties had been discovered in the early 1920s, the different characteristics associated with different molecular-weight fractions of heparin were only recognized in the late 1970s. Tinzaparin is a low-molecular-weight heparin (LMWH) produced by heparinase digestion of heparin. Preclinical research on tinzaparin established that there were no differences in the antithrombotic activity compared with heparin. Clinical studies evaluating tinzaparin vs. standard heparin for thromboprophylaxis of deep-vein thrombosis in general and orthopedic surgery found that tinzaparin was as effective as standard heparin. Tinzaparin was also evaluated vs. standard heparin in the treatment of acute proximal vein thrombosis; time-to-event curves suggested that this LMWH could be more effective than standard heparin.

Requirement for Anticoagulant Heparan Sulfate in the Fibroblast Growth Factor Receptor Complex

A divalent cation-dependent association between heparin or heparan sulfate and the ectodomain of the fibroblast growth factor (FGF) receptor kinase (FGFR) restricts FGF-independent trans-phosphorylation between self-associated FGFR and determines specificity for and mediates binding of activating FGF. Here we show that only the fraction of commercial heparin or rat liver heparan sulfate which binds to immobilized antithrombin formed an FGF-binding binary complex with the ectodomain of the FGFR kinase. Conversely, only the fraction of heparin that binds to immobilized FGFR inhibited Factor Xa in the presence of antithrombin. Only the antithrombin-bound fraction of heparin competed with (3)H-heparin bound to FGFR in absence of FGF, whereas both antithrombin-bound and unretained fractions competed with radiolabeled heparin bound independently to FGF-1 and FGF-2. The antithrombin-bound fraction of heparin was required to support the heparin-dependent stimulation of DNA synthesis of endothelial cells by FGF-1. The requirement for divalent cations and the antithrombin-binding motif distinguish the role of heparan sulfate as an integral subunit of the FGFR complex from the wider range of effects of heparan sulfates and homologues on FGF signaling through FGFR-independent interactions with FGF.

Thrombin-induced platelet activation is inhibited by high- and low-molecular-weight heparin.

Thrombin binds to platelet glycoprotein Ib (Gp Ib), and this interaction contributes to platelet activation. Thrombin ligation to Gp Ib was recently shown to be inhibited by heparin, thus raising the hypothesis, investigated in this article, that heparin might inhibit thrombin-induced platelet activation. Aggregation of gel-filtered platelets by 1 nmol/L thrombin was reduced by both high-molecular-weight (MW) (14 500-Da) and low-MW (4500-Da) heparin, with IC50 values of 1.65+/-0.26 and 5.13+/-0.8 micromol/L, respectively. Homogeneous-MW fractions (16 000- to 13 000-Da range) were used to evaluate the heparin effect on intracytoplasmic calcium release by thrombin. Calcium mobilization by 1 nmol/L thrombin was reduced as a function of heparin concentration, and the inhibitory effect was correlated to the MW of heparin fractions (IC50 values were 1.9+/-0.39, 6.07+/-0.83, and 14. 8+/-0.43 micromol/L for 16 000-, 9000-, and 3000-Da heparin, respectively). Platelet aggregation and calcium mobilization by ADP and by the thrombin receptor-activating peptide were not affected by heparin. The activation of Gp Ib-depleted platelets by alpha-thrombin was not inhibited by heparin. Moreover, platelet stimulation by heparin binding site phosphopyridoxylated thrombin, which has a severe impairment of Gp Ib ligation, was not affected by heparin. Finally, heparin did not interfere with the hydrolysis by thrombin of the protease-activated receptor 1. These results demonstrated that heparin, by inhibiting the thrombin-Gp Ib interaction, is able to interfere with thrombin-induced platelet activation. The extent of the inhibitory effect is directly related to the MW of heparin fractions.

Effects of heparin on vascular dysfunction in diabetic rats.

1. Treatment with heparin has beneficial effects in diabetic nephropathy. The occurrence of increased urinary albumin excretion in diabetic patients reflects general vascular dysfunction, including increased transcapillary permeability of macromolecules. The aim of the present study was to evaluate the effects of heparin on vascular dysfunction in diabetic rats. 2. Male Sprague-Dawley rats were used in two studies. Diabetes was induced by 65 mg/kg, i.v., streptozotocin. In one study, diabetic rats were dosed subcutaneously with different heparin fractions for 8 months and the transcapillary escape rate of albumin (TERalb) was measured in anaesthetized animals. In the other study, heparin was given for 6 weeks, followed by tissue albumin clearance measurements in awake rats. Normal and diabetic rats receiving saline served as controls. 3. Blood glucose did not differ among the diabetic groups and ranged from 22 to 26 mmol/L. The mean (+/- SD) TERalb was increased by diabetes compared with values in normal rats (17.5 +/- 3 vs 14.1 +/- 3.3%/h, respectively). Neither unfractionated nor low molecular weight heparin significantly affected this increase. [131I]-Albumin clearance was significantly increased in diabetic rats in the eye, skin and skeletal muscle tissues compared with normal rats (0.17-0.40 vs 0.1-0.23 microL plasma/g per min). Low molecular weight heparin treatment did not affect the increased organ albumin clearance. 4. In conclusion, heparin treatment does not affect diabetes-induced vascular dysfunction as expressed by increased TERalb and clearance of albumin in rats.

LMW-HEPARIN/-HEPARINOID ANTI-XA ACTIVITY OBTAINED BY HMT (CELLITE) AND ACT (CELLITE, KAOLIN) PROCEDURES

S187 The new heparin management test (HMT, dry chemistry) has been proved a reliable point-of-care system for monitoring unfractioned heparins during cardiopulmonary bypass (CPB) [1]. The HMT has been also shown to correlate well with the anti-Xa activity [1], which is regarded as a reference for the determination of plasma heparin levels. However, chromogen-measured anti-Xa activity does not only take into account the high-molecular-weight fractions of heparin, but also depends on the low-molecular-weight (LMW) fraction. The goal of the present study was to assess whether the optical HMT method reveals linearity with LMWH-anti-Xa activity, and to compare the results to the established mechanical ACT (activated clotting time) procedures (surface activator cellite or kaolin). METHODS: With approval of the local ethics comittee and written informed consent, 5 patients scheduled for elective CPB participated. Defined LMWH anti-Xa activities were achieved by spiking whole blood samples of each patient with 0.25 U/mL (range: 0 - 3 U/mL) of the heparinoid orgaran (NV Organon, OSS). Clotting times (sec) were obtained simultaneously with the cellite and kaolin ACT (Hemochron; ITC, Edison, NJ, USA) (spiked whole blood; n = 24), and the new cellite HMT method (Cardiovascular Diagnostics Inc., Raleigh, NC, USA) (citrated whole blood; n = 12). All samples were analyzed at 13 set points immediately after spiking (0-30 U/mL; duplicates, n = 72). RESULTS: The ACT and HMT values (sec) at defined concentrations of orgaran (U/mL) are shown below (mean values, U-test). (Figure 1)Figure 1CONCLUSION: Recently, in contrast to the ACT, the new cellite HMT has been shown to be independent of potential errors (e.g., due to aprotinin) [1]. Presently, the HMT additionally correlated well with LMWH-anti-Xa activity over a wide range, whereas the cellite ACT appears to be rather uninfluenced by this heparin fraction (i.e., not suitable). The molecular mechanism of this finding, however, remains a matter of speculation. Further studies will have to prove whether the HMT may develop routine on-line monitoring for high levels of both LMW-heparins and - heparinoids (e.g., orgaran), too, as occuring in invasive cardiology, hemodialysis, and CPB (including patients with heparin-induced thrombocytopenia type II).

O9 The anti-Xa activities of high and low affinity fractions of heparin

O9 The anti-Xa activities of high and low affinity fractions of heparin

Case studies in therapeutics: warfarin resistance and inefficacy in a man with recurrent thromboembolism, and anticoagulant‐associated priapism

Case studies in therapeutics: warfarin resistance and inefficacy in a man with recurrent thromboembolism, and anticoagulant‐associated priapism

Improved microsurgical anastomotic patency with low molecular weight heparin.

Blood flow to a free flap may be impaired by thrombotic occlusion at the anastomosis or by microemboli occluding microvessels. The purpose of this study was to test the hypothesis that unfractionated heparin (UFH) or low molecular weight fractions of heparin (LMWH) could improve both the patency of microvascular anastomoses and microcirculatory perfusion. Sixty-six rats underwent orthotopic elevation of 3- x 10-cm epigastric free flaps. Animals received a single injection of either vehicle, UFH or LMWH, prior to microvascular clamp application and pedicle division. Anastomotic patency and tissue survival area were assessed on postoperative day 7. Anastomotic patency was significantly improved in both the UFH and LMWH groups. Total tissue survival area in those flaps with anastomotic patency was significantly improved in the UFH and the LMWH groups. Although both UFH and LMWH significantly elevated activated partial thromboplastin times (APTT) and anti activated clotting factor X (anti-Xa) activity over controls, UFH had its greatest effect on APTT, and LMWH on anti-factor Xa activity. Hematomas developed only in the UFH group. Thus, although both UFH and LMWH improved microcirculatory perfusion, as indicated by increased flap survival, only LMWH improved anastomotic patency while minimizing hemorrhage.

A fraction of high molecular weight heparin has high affinity for vitronectin

A fraction of high molecular weight heparin has high affinity for vitronectin

Suppression of Endothelin-1 Production by Heparin Disaccharides. † 1652

Endothelin-1 (ET-1), the most potent vasoconstrictor peptide in nature, is involved in systemic and pulmonary hypertension and in closure of the ductus arteriosus in neonates. Heparin, a heterogeneous glycosaminoglycan, suppresses the production of ET-1 in human unbilical vein cultured endothelial cells(HUVEC). We have shown that the most negatively charged fractions of heparin have the highest ET-1-suppressive activity. This effect of heparin is independent of size, down to 2,400 daltons (octasaccharide).

Inhibition of antigen-induced airway hyperresponsiveness by ultralow molecular-weight heparin.

Unfractionated heparin (UF-heparin) has been shown to prevent antigen-induced airway hyperresponsiveness (AHR), but it is ineffective when administered after the antigen challenge. We hypothesized that the failure of UF-heparin to modify postantigen AHR might depend on molecular weight. We therefore studied the effects of UF-heparin and three low-molecular-weight heparin fractions (medium-molecular-weight heparin [MMWH]; low-molecular-weight heparin [LMWH]; and ultralow-molecular-weight heparin [ULMWH]) on antigen-induced AHR and histamine release in bronchoalveolar lavage fluid (BALF). Specific lung resistance (SRL) was measured in 20 allergic sheep before, immediately after, and up to 2 h after challenge with Ascaris suum antigen. Airway responsiveness was expressed as the cumulative provocative dose of carbachol, in breath units, that increased SRL by 400% (PD400). PD400 was determined before and 2 h after antigen, both without and after treatment with aerosolized UF-heparin (1,000 U/kg) and various heparin fractions (0.04 mg/kg to 5 mg/kg) administered after the antigen challenge. Inhaled UF-heparin (n = 4), MMWH (n = 4), and LMWH (n = 6) failed to modify postantigen AHR when administered after the challenge. Only ULMWH (n = 6) inhibited postantigen AHR in a dose-dependent manner (percent protection ranged from 31% to 139%). In eight additional sheep, histamine in BALF was measured with a radioimmunoassay (RIA) before and after the segmental antigen challenge, without and after pretreatment with inhaled UF-heparin, LMWH, or ULMWH. Inhaled UF-heparin and LMWH inhibited antigen-induced histamine release as measured in BALF by 81% and 75%, respectively; whereas ULMWH was ineffective in this respect. We conclude that: (1) modification of antigen-induced AHR by fractionated heparins is molecular-weight dependent; and (2) only ULMWH attenuates AHR when administered after antigen challenge, via an unknown mast-cell-independent action.

Inhibition of allergic airway responses by inhaled low-molecular-weight heparins: molecular-weight dependence.

Inhaled heparin prevents antigen-induced bronchoconstriction and inhibits anti-immunoglobulin E-mediated mast cell degranulation. We hypothesized that the antiallergic action of heparin may be molecular weight dependent. Therefore, we studied the effects of three different low-molecular-weight fractions of heparin [medium-, low-, and ultralow-molecular-weight heparin (MMWH, LMWH, ULMWH, respectively)] on the antigen-induced acute bronchoconstrictor response (ABR) and airway hyperresponsiveness (AHR) in allergic sheep. Specific lung resistance was measured in 22 sheep before and after airway challenge with Ascaris suum antigen, without and after pretreatment with inhaled fractionated heparins at doses of 0.31-5.0 mg/kg. Airway responsiveness was estimated before and 2 h postantigen as the cumulative provocating dose of carbachol in breath units that increased specific lung resistance by 400%. All fractionated heparins caused a dose-dependent inhibition of ABR and AHR. ULMWH was the most effective fraction, with the inhibitory dose causing 50% protection (ID50) against ABR of 0.5 mg/kg, whereas ID50 values of LMWH and MMWH were 1.25 and 1.8 mg/kg, respectively. ULMWH was also the most effective fraction in attenuating AHR; the ID50 values for ULMWH, LMWH, and MMWH were 0.5, 2.5, and 4.7 mg/kg, respectively. These data suggest that 1) fractionated low-molecular-weight heparins attenuate antigen-induced ABR and AHR; 2) there is an inverse relationship between the antiallergic activity of heparin fractions and molecular weight; and 3) ULMWH is the most effective fraction preventing allergic bronchoconstriction and airway hyperresponsiveness.

Review of pharmacodynamics, pharmacokinetics, and therapeutic properties of sulodexide.

Due to various side effects of heparin, such as heparin-induced thrombocytopenia type I or type II, alternative anticoagulants are in clinical development to optimize the anticoagulant regimes in patients requiring low or high anticoagulation dosages. Sulodexide is a highly purified preparation containing a fast-moving heparin fraction as well as dermatansulfate. The pharmacological effects of sulodexide differ substantially from unfractionated heparin and are mainly characterized by a prolonged half-life and reduced effect on global coagulation and bleeding parameters. The lipolytic activity of sulodexide is increased in comparison to unfractionated heparin. Clinical studies demonstrate the safety and efficacy of sulodexide. Specially, oral administration leads to fibrinolytic activities in contrast to oral administration of other glycosaminoglycans. Thus, sulodexide releases tissue plasminogen activator and decreases fibrinogen levels as well as HDL and total cholesterol levels and blood viscosity. Clinical efficacy is demonstrated in peripheral arterial disease, cardiovascular events, in postphlebitic syndrome and on albuminuria in nephropathy.

Influence of Molecular Weight, Protein Core and Charge of Native Heparin Fractions on Pulmonary Artery Smooth Muscle Cell Proliferation

Heparin macromolecules have been shown to inhibit cultured pulmonary artery smooth muscle cell proliferation in vitro and prevent hypoxic vascular remodeling in vivo. In an attempt to understand the structural determinants of heparin's antiproliferative properties, we have fractionated an antiproliferative preparation of commercial heparin into low and high molecular weight fractions. Then the high molecular weight heparin fraction was further fractionated on a DEAE-cellulose column by charge density eluting with 0 - 1 M NaCl linear gradient. The heparin protein peptides were both removed and isolated. These heparin fractions were assayed for antiproliferative effects on cultured bovine pulmonary artery smooth muscle cells. No appreciable differences were found among high and low molecular weight heparin fractions The core peptides showed no antiproliferative activity. However, higher charge density fraction was less antiproliferative.

Structural and functional modifications of bovine trypsin by heparins: Influence of heparin molecular mass and structure

Heparins with different structures, charge density and molecular mass were evaluated for their capacity to induce structural and functional alterations of bovine trypsin in a low ionic strength buffer (20 mM Tris-HCl pH 7.4). Unfractionated heparin, and slow and fast moving heparin species increased the fluorescence peak emission of trypsin to the same extent (about +40.0%), whilst partially desulfated and re-N-acetylated heparin with a charge density of 1.47 modified the fluorescence at 330 nm by about +27% and natural heparan sulfate with a sulfate-to-carboxyl ratio <1 by about +13%. Heparin fractions with narrow polydispersity and the same charge density (produced by chemical depolymerization in the presence of free radicals and further gel-permeation chromatography) having molecular mass lower than about 6000 interact with trypsin to a less extent, even though fractions with molecular mass of about 4500 and 3600 partially retain this property. No modification of fluorescence peak emission of trypsin with heparin was appreciable when the ionic strength of the buffer was increased to 0.3 mM NaCl. An altered ability to reduce cytochrome c was observed for heparins of different charge density; fragments with molecular mass lower than approximately 4000 were also unable to produce superoxide. Trypsin was degraded into fragments by heparin and derivatives after 3 h incubation at 37°C. After electrophoresis in polyacrylamide-gels the trypsin bands disappeared and fragments with lower molecular mass were more evident. This effect depended on the molecular mass of heparin, and was more evident for unfractionated heparin and for a heparin fraction with a molecular mass of 7820. The esterolytic activity of trypsin was inhibited to the same extent by heparin derivatives of various structure and charge density while activity undermet minor changes in the presence of heparin fractions of M r lower than 4000.

Low molecular weight heparin: biochemistry, pharmacology, perioperative prophylaxis regimens, and guidelines for regional anesthetic management.

Low molecular weight heparin: biochemistry, pharmacology, perioperative prophylaxis regimens, and guidelines for regional anesthetic management.

Inhibition of antigen-induced acute bronchoconstriction, airway hyperresponsiveness, and mast cell degranulation by a nonanticoagulant heparin: comparison with a low molecular weight heparin.

Inhaled heparin prevents antigen-induced bronchoconstriction and inhibits anti-IgE-mediated mast-cell degranulation. We hypothesized that the antiallergic action of heparin may be dependent on molecular weight and related to its nonanticoagulant properties. Therefore, in the present investigation we studied the effects of a nonanticoagulant fraction of heparin (LA-heparin) on antigen-induced bronchoconstriction, airway hyperresponsiveness (AHR), and mast-cell degranulation, and compared its antiallergic activity with that of a low molecular weight heparin (LMW-heparin, fragmin). Specific lung resistance (SRL) was measured in 15 sheep before, immediately after, and serially for as long as 2 h after airway challenge with Ascaris suum antigen, without and after pretreatment with inhaled fractionated heparins at doses of 2.5 and 5 mg/kg. Airway responsiveness was estimated before, and 2 h after antigen as the cumulative provocating dose (PD400) of carbachol in breath units, which increased SRL by 400% (one breath unit was defined as one breath of 1% carbachol solution). LA-heparin caused a dose-dependent inhibition of antigen-induced bronchoconstriction, and a 5-mg/kg nebulized dose caused a 67% inhibition of allergic bronchoconstriction, whereas a 2.5-mg/kg dose was ineffective (20% inhibition). Inhaled fragmin was more potent than LA-heparin, as shown by 84% (2.5 mg/kg) and 82% (5 mg/kg) inhibition of allergic bronchoconstriction. Fragmin (5 mg/kg) also attenuated the postantigen AHR, whereas LA-heparin was ineffective. In vitro, preincubation with both LA-heparin and fragmin inhibited the anti-IgE-induced degranulation of rat peritoneal mast-cells in a dose-dependent fashion. LA-heparin was fourfold more potent than fragmin, with IC50 of 80 and 320 microg/ml, respectively. These data suggest that: (1) fractionated heparins attenuate antigen-induced acute bronchoconstriction, (2) nonanticoagulant fractions mediate the antiallergic activity of inhaled heparin, and (3) antiallergic activity of nonanticoagulant heparin and LMW-heparin may be related to prevention of mast-cell degranulation.