Abstract 28: Serial in Vivo Imaging of Thrombus Inflammation Predicts Venous Thrombus Resolution

2012 ◽  
Vol 32 (suppl_1) ◽  
Author(s):  
Chase W Kessinger ◽  
Jason R McCarthy ◽  
Charles P Lin ◽  
Peter K Henke ◽  
Farouc A Jaffer
Keyword(s):  
Fluorescence Microscopy ◽  
In Vivo Imaging ◽  
Macrophage Infiltration ◽  
Intravital Fluorescence Microscopy ◽  
Targeted Nanoparticles ◽  
Macrophage Activity ◽  
Vein Thrombosis ◽  
Deep Vein ◽  
In Vivo Assessment

Introduction: Inflammation following deep vein thrombosis (DVT) critically modulates thrombus resolution. However it is unknown whether the degree of inflammation in vivo predicts the magnitude of thrombus resolution. Here, we utilize serial multichannel intravital fluorescence microscopy (IVFM) to spatially map and quantify macrophage infiltration in thrombus, and then to determine the degree of thrombus resolution in vivo . Methods: C57Bl/6 mice (n=5) underwent topical ferric chloride injury to induce femoral venous thrombosis. On day 3, mice were i.v. injected with macrophage-targeted nanoparticles (CLIO-AF555, ex/em 555/565nm). On day 4, survival IVFM of thrombus macrophages was performed. In addition, venography was concomitantly performed (pre-injection of FITC-dextran 45 minutes before IVFM, MW 2000 kD, 490/520nm) to quantify DVT area and length in vivo at day 4 and 6. IVFM image analysis was performed using ImageJ. Mid-luminal z-stacks (40 μm thickness) were analyzed. Thrombus ROIs were outlined using FITC-dextran generated angiograms, and used calculating macrophage target-to-background ratios (TBR) and thrombus area and length. Contralateral sham femoral veins were imaged as controls. Imaging was followed by histopathology and fluorescence microscopy of the cryosectioned tissue. Results: At day 4, the baseline femoral DVT area and length were 0.22±0.08mm 2 and 1.29±0.16mm (mean±sd). The day 4 macrophage activity was noted to be heterogeneous with peripheral infiltration of the thrombus, and occasionally with some deeper penetration into the thrombus interior. At day 6, the thrombus area and length were reduced compared to day 4 (p<0.005 for both). The day 4 thrombus macrophage TBR significantly predicted the degree of thrombus reduction (correlation of initial macrophage TBR and Δthrombus area, r=0.97, p=0.001). Conclusion: This serial in vivo assessment of murine DVT resolution demonstrates that degree of macrophage infiltration at day 4 predicts the magnitude of thrombus resolution at day 6. Molecular imaging of DVT inflammation in vivo could provide new insights into DVT resolution and the development of the post-thrombotic syndrome, and also in evaluating inflammatory-modulating therapies for improved DVT resolution.

In Vivo Porcine Aged Deep Vein Thrombosis Model for Testing Ultrasound-based Thrombolysis Techniques

2021 ◽  
Author(s):  
Greyson E. Stocker ◽  
Jiaqi Shi ◽  
Kimberly Ives ◽  
Adam D. Maxwell ◽  
Paul A. Dayton ◽  
...  
Keyword(s):  
Deep Vein Thrombosis ◽  
Vein Thrombosis ◽  
Thrombosis Model ◽  
Deep Vein

Increase of Thrombin-Antithrombin III (TAT) Complex Plasma Levels in Thromboembolic Diseases during Thrombolysis

1987 ◽  
Author(s):  
R Seitz ◽  
G Pratorius ◽  
R Blanke ◽  
B B Strauer
Keyword(s):  
Thrombolytic Therapy ◽  
Plasma Levels ◽  
Vein Thrombosis ◽  
Complex Plasma ◽  
In Vitro Effects ◽  
Immuno Assay ◽  
Enzyme Immuno Assay ◽  
Deep Vein

Recently an enzyme immuno assay of thrombin-antithrombinlll complex (TAT) plasma levels was developed by PELZER et al. (Thromb. Haemost. 54:24,1985). This test appears to be useful in the detection of intravasal thrombin generation, since all of 17 patients (pts.) with pulmonary embolism and 15 of 16 pts. with deep vein thrombosis (DVT) showed elevated values above 3 ng/ml.In 9 pts. with acute myocardial infarction (AMI) the TAT levels increased significantly (p 0.001) 3 to 6 hours after thrombolytic therapy with 1.5 million units streptokinase (SK) over 30 minutes. A concomitant increase of fibrinopeptide A (FPA) levels (p=0.048) was observed. In contrast, 8 AMI pts. treated with heparin showed an insignificant increase of TAT and FPA. In 7 DVT pts. the TAT levels rose significantly (p 0.001) within 6 hours after start of urokinase (UK) infusion, while the FPA levels were enhanced prior to treatment and showed no further increase.In order to assess the in vitro effects of SK and UK on TAT levels, clots obtained by recalcification of citrated plasma were incubated in heparin (2 units/ml) plasma. An increase of TAT occurred after addition of SK or UK, which was less pronounced when the clots were rinsed extensivly or squeezed before incubation. When SK or UK were added to plasma in the absence of a clot, still a small increase of TAT occurred which was absent in saline controls.The data suggest that SK and UK action is associated with the generation of TAT complexes. In vivo, thrombin or thromboplastic material might be released by enhanced "wash out" from the recanalized coronary artery or from the reperfused in-farcted myocardium. Thrombin might also be released from binding sites on fibrin clots or fibrinogen. It is conceivable that these findings contribute to the understanding of reocclusion of infarct vessels after thrombolytic therapy. This points to the importance of careful anticoagulation in patients receiving thrombolytic therapy.

Phase I Study of an Immunomodulatory Thalidomide Analog, CC-4047, in Relapsed or Refractory Multiple Myeloma

2004 ◽  
Vol 22 (16) ◽  
pp. 3269-3276 ◽  
Author(s):  
S.A. Schey ◽  
P. Fields ◽  
J.B. Bartlett ◽  
I.A. Clarke ◽  
G. Ashan ◽  
...  
Keyword(s):  
T Cell ◽  
Primary Melanoma ◽  
Maximum Tolerated Dose ◽  
T Cell Subsets ◽  
Serious Adverse Event ◽  
Vein Thrombosis ◽  
Monocytes And Macrophages ◽  
Clinical Responses ◽  
Deep Vein

Purpose To assess the safety, efficacy, and immunomodulatory effects of CC-4047 (Actimid; Celgene, San Diego, CA) in patients with relapsed or refractory myeloma. Patients and Methods Twenty-four relapsed or refractory patients were treated with a dose-escalating regimen of oral CC-4047. Clinical responses and adverse effects were identified, and peripheral T-cell subsets, serum cytokines, and proangiogenic factors were evaluated. Results CC-4047 was tolerated with no serious nonhematologic adverse events. All patients were eligible for analysis. Toxicity criteria during the initial 4 weeks of study were used to define the maximum-tolerated dose (MTD). During this period, one patient withdrew with a deep vein thrombosis (DVT) probably caused by an undiagnosed primary melanoma with lymphadenopathy in the groin, one patient withdrew because of progressive disease (PD), and three patients discontinued with neutropenia. Nineteen of 24 patients continued on treatment beyond 4 weeks to PD or development of a serious adverse event. Three further patients developed a DVT at 4, 9, and 11 months. Treatment resulted in a greater than 25% reduction in paraprotein in 67% of patients, 13 patients (54%) experienced a greater than 50% reduction in paraprotein, and four (17%) of 24 patients entered complete remission. The MTD was 2 mg/d. All patients showed increased CD45RO expression on CD4+ and CD8+ cells, with a concomitant decrease in CD45RA+ cells. CC-4047 treatment was associated with significantly increased serum interleukin (IL)-2 receptor and IL-12 levels, which is consistent with activation of T cells and monocytes and macrophages. Conclusion This study demonstrates the safety and efficacy of CC-4047. The MTD of CC-4047 orally was 2 mg/d. This is the first report demonstrating in vivo T-cell costimulation by this class of compound, supporting a potential role for CC-4047 as an immunostimulatory adjuvant treatment.

In vivo imaging of hepatic excretory function in the rat by fluorescence microscopy

2012 ◽  
Vol 5 (7) ◽  
pp. 571-581 ◽  
Author(s):  
Peter Recknagel ◽  
Ralf A. Claus ◽  
Ute Neugebauer ◽  
Michael Bauer ◽  
Falk A. Gonnert
Keyword(s):  
Fluorescence Microscopy ◽  
In Vivo Imaging ◽  
Excretory Function

scholarly journals A general method to fine-tune fluorophores for live-cell and in vivo imaging

2017 ◽  
Author(s):  
Jonathan B. Grimm ◽  
Anand K. Muthusamy ◽  
Yajie Liang ◽  
Timothy A. Brown ◽  
William C. Lemon ◽  
...  
Keyword(s):  
Fluorescence Microscopy ◽  
In Vivo Imaging ◽  
Chemical Properties ◽  
Live Cell ◽  
Live Cells ◽  
Fine Tune ◽  
General Method ◽  
And Chemical Properties

ABSTRACTPushing the frontier of fluorescence microscopy requires the design of enhanced fluorophores with finely tuned properties. We recently discovered that incorporation of four-membered azetidine rings into classic fluorophore structures elicits substantial increases in brightness and photostability, resulting in the ‘Janelia Fluor’ (JF) series of dyes. Here, we refine and extend this strategy, showing that incorporation of 3-substituted azetidine groups allows rational tuning of the spectral and chemical properties with unprecedented precision. This strategy yields a palette of new fluorescent and fluorogenic labels with excitation ranging from blue to the far-red with utility in live cells, tissue, and animals.

scholarly journals Advanced fluorescence microscopy for in vivo imaging of neuronal activity

Optica ◽  
2019 ◽  
Vol 6 (6) ◽  
pp. 758 ◽  
Author(s):  
Giuseppe Sancataldo ◽  
Ludovico Silvestri ◽  
Anna Letizia Allegra Mascaro ◽  
Leonardo Sacconi ◽  
Francesco Saverio Pavone
Keyword(s):  
Fluorescence Microscopy ◽  
Neuronal Activity ◽  
In Vivo Imaging

A Comparative Study of Prothrombinase and Thrombin Inhibitors in a Novel Rabbit Model of Non-Occlusive Deep Vein Thrombosis

1994 ◽  
Vol 71 (03) ◽  
pp. 357-362 ◽  
Author(s):  
Stan Hollenbach ◽  
Uma Sinha ◽  
Pei-Hua Lin ◽  
Kathy Needham ◽  
Lisa Frey ◽  
...  
Keyword(s):  
Deep Vein Thrombosis ◽  
Ex Vivo ◽  
Rabbit Model ◽  
Thrombin Inhibitors ◽  
Constant Infusion ◽  
Chloromethyl Ketone ◽  
Vein Thrombosis ◽  
Prothrombinase Complex ◽  
Deep Vein

SummaryA quantitative and non-occlusive deep vein thrombosis model was developed in rabbits. We used this model to test the antithrombotic activity of the prothrombinase complex inhibitors factor rXai and its chemical analog glutamyl-glycyl-arginyl chloromethyl ketone inactivated human factor Xa (EGR-Xai), along with the thrombin inhibitors D-phenylalanyl-prolyl-arginyl chloromethyl ketone (PPACK) and heparin. Dose dependent effects of the inhibitors during constant infusion were monitored. Measurements included thrombus weights, hemostatic parameters and both cuticle and ear bleeding times. In this model, factor rXai and EGR-Xai had comparable in-vivo efficacy, and showed 80%-93% inhibition at plasma levels of 6.5 nM (rXai) and 8 nM (EGR-Xai). Effects on ex-vivo clotting times varied among the inhibitors. At 80-100% thrombus inhibition, factor rXai and EGR-Xai had no statistically significant effect, while PPACK extended thrombin clotting time (TCT) times 2.3-fold, and heparin prolonged both activated partial thromboplastin time (APTT), prothrombin time (PT) and TCT ex-vivo clotting times 6.9-, 1.2-, and 7-fold respectively. At these dosages, cuticle and ear bleeding times were prolonged for all inhibitors and showed increases of 177%-389% (cuticle) and 45%-129% (ear). Our results demonstrate that direct inhibition of prothrombinase complex assembly is effective in arresting venous thrombosis.

Abstract 52: Statin Therapy Accelerates Venous Thrombus Resolution: Assessment In Stasis And Chemical Injury Induced Thrombosis

2013 ◽  
Vol 33 (suppl_1) ◽  
Author(s):  
Chase W Kessinger ◽  
Jin Won Kim ◽  
Brian Thompson ◽  
Martin Sillesen ◽  
Tetsuya Hara ◽  
...  
Keyword(s):  
Statin Therapy ◽  
Platelet Activation ◽  
Femoral Vein ◽  
Plasminogen Activator Inhibitor ◽  
Plasminogen Activator Inhibitor 1 ◽  
Chemical Injury ◽  
Vein Thrombosis ◽  
Activity Measurements ◽  
Deep Vein

Introduction In deep vein thrombosis (DVT), both the induced inflammation response and fibrinolytic capacity are critical in resolving experimental DVT. Unresolved, obstructing thrombi can lead to the burdensome and costly sequelae of the post-thrombotic syndrome. Recently, statins have shown to reduce the incidence of DVT (JUPITER trial). In this study, we investigated the in vivo effects of statin therapy on resolution of already formed DVT using serial intravital microscopy (IVM) imaging and thrombus mass measurements. Methods DVT was induced in C57Bl/6J male mice (N=190) by topical ferric chloride of the femoral vein (chemical injury) or ligation of the IVC (stasis). One day after DVT induction, mice started treatment of atorvastatin (1.14mg/kg), mevastatin (10 mg/kg) or PBS by once daily oral gavage. Serial IVM was preformed at days 2, 4 and 6 utilizing FITC-dextran (ex/em 490/520nm), CLIO-AF555 (ex/em 555/565nm), and MMPSense680 (ex/em 680/700nm) imaging agents for thrombus architecture, macrophage content and MMP activity measurements. Thrombus masses were collected at days 4, 7, 10 in stasis DVT mice and combination statin/enoxaparin (10mg/kg/d, s.c.) were also compared to statin therapy. Platelet activation and coagulation were analyzed using aggregometry and thromboelastography on whole-blood. Results We found that statin therapy reduced DVT burden and inflammation in both chemical- and stasis-induced established DVT up to 10 days compared to control (p<0.05). Serial IVM imaging of statin animals showed an increase in DVT resolution from day 2 to 4 (Δlength -21± 6.0 vs. control -7.1± 5.6%, p<0.01), and decreased inflammation compared to controls (p<0.001). Inflammation was also found to predict the extent of DVT resolution(r=0.84). Statin therapy also reduced thrombus plasminogen activator inhibitor-1, tissue factor, and Mac-3 levels, as well as reducing platelet activation and clot strength(p<0.05). Conclusion Statin therapy accelerates DVT resolution despite reducing thrombus related inflammation. The net reduction of thrombus burden tracked with statin-mediated favorable effects on fibrinolysis, coagulation, and platelet function. These results support statins as a translatable therapy to improve the resolution of DVT.

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