scholarly journals Estimation of corneal thickness ex vivo and in vivo at instillation of photosensitizer solutions in different modifications of UV corneal crosslinking

2021 ◽  
Vol 25 (2) ◽  
pp. 96-105
Author(s):  
M. M. Bikbov ◽  
A. R. Khalimov ◽  
V. K. Surkova ◽  
G. M. Kazakbaeva
Keyword(s):  
Uv Irradiation ◽  
Ex Vivo ◽  
Corneal Thickness ◽  
Radiation Energy ◽  
Rational Approach ◽  
Initial State ◽  
Corneal Crosslinking ◽  
First Case ◽  
Intraoperative Control

Objectives: to evaluate the effectiveness of photosensitizer solutions in the experiment in different modifications of standard UV corneal crosslinking. Materials and Methods. Experiments were performed on 32 enucleated porcine eyes ex vivo and 12 rabbits (24 eyes) in vivo, divided into 4 groups depending on instillations of the photosensitizer solutions: Dextralink, Ribolink, Hypolink and Riboflavin. Evaluation of corneal saturation was performed using two modifications of standard UV CXL: in the first case, instillation of the photosensitizer solution was performed during the entire crosslinking procedure (30 min - saturation and 30 min - ultraviolet irradiation); in the second case, instillations lasted only for the first 30 minutes, the precorneal riboflavin film was removed, and instillation of solution was not performed. Results and Discussion. Instillation of photosensitizer solutions within 60 minutes showed that Dextralink significantly reduced the thickness of the cornea by about 24 % ex vivo and 21 % in vivo, while Hypolink, on the contrary, caused its increase by 9 % ex vivo and 23 % in vivo, respectively. The use of Ribolink and Riboflavin did not change the linear parameters of the cornea during the entire follow-up period. The results of the 2nd series of studies showed that effects of solutions on the cornea during their instillation for 30 minutes were generally preserved during the entire observation period up to 60 minutes. In the group where Dextralink and Hypolink solutions were used, there was a slight tendency to normalization of the initial parameters of the cornea. Conclusion. Safe and effective implementation of UV crosslinking of the cornea is possible on the basis of a rational approach to the performing the stages of stroma saturation with photosensitizer solutions, depending on the initial state of the cornea: Dextralink is recommended for a thickness of more than 450 m, Ribolink - at 400-450 m, Hypolink - from 350 to 400 m. Halting of instillations on the saturated stroma during UV irradiation is advisable to be accompanied by intraoperative control of its thickness. When performing a standard crosslinking technique, where instillations of photosensitizer accompany the stage of UV irradiation of the cornea, it is necessary to take into account the presence of a precorneal film that can absorb some of the radiation energy. The peculiarity of this crosslinking technique can be considered as a potential way to protect intraocular tissues from excessive exposure to UV radiation.

scholarly journals Identification of lymphatic pathway involved in the spread of bladder cancer: Evidence obtained from fluorescence navigation with intraoperatively injected indocyanine green

2013 ◽  
Vol 7 (5-6) ◽  
pp. 322 ◽  
Author(s):  
Shogo Inoue ◽  
Hiroaki Shiina ◽  
Yozo Mitsui ◽  
Hiroaki Yasumoto ◽  
Akio Matsubara ◽  
...  
Keyword(s):  
Indocyanine Green ◽  
Radical Cystectomy ◽  
Fluorescence Intensity ◽  
Ex Vivo ◽  
Lymphatic Vessels ◽  
Rational Approach ◽  
Lymphatic Channel ◽  
Internal Iliac ◽  
Lymphatic Pathway

Introduction: We identify lymphatic vessels draining from the bladderby using fluorescence navigation (FN) system.Methods: In total, 12 candidates for radical cystectomy and pelviclymph node dissection (PLND) were included in this study. Afteran indocyanine green (ICG) solution was injected into the bladderduring radical cystectomy, lymphatic vessels draining from thebladder were analyzed using a FN system. PLND was based onthe lymphatic mapping created from the FN measurements (in vivoprobing) in the external iliac, obturator and internal iliac regions;after PLND, the fluorescence of the removed lymph nodes (LNs)was analyzed on the bench (ex vivo probing).Results: There were no patients with complications associated withthe intravesical ICG injection. A lymphatic pathway along inferiorvesical vessels to internal iliac LNs was clearly illustrated in 7 cases.Under in-vivo probing, the fluorescence intensity of internal iliacnodes was greater than that of external iliac or obturator nodes.Under ex-vivo probing, the fluorescence intensity of internal iliacand obturator nodes was greater than that of external iliac nodes.Conclusions: Using an FN system after injecting ICG during a radicalcystectomy operation is a safe and rational approach to detectingthe lymphatic channel draining from the bladder.

scholarly journals Transepithelial corneal cross-linking assisted by two continuous cycles of iontophoresis for progressive keratoconus in adults: retrospective 5-year analysis

2020 ◽  
Vol 259 (1) ◽  
pp. 239-246
Author(s):  
Huping Wu ◽  
Shunrong Luo ◽  
Xie Fang ◽  
Xumin Shang ◽  
Zhiwen Xie ◽  
...  
Keyword(s):  
Corneal Thickness ◽  
Anterior Segment ◽  
Long Term Effects ◽  
Corneal Crosslinking ◽  
Endothelial Cell Count ◽  
Corneal Confocal Microscopy ◽  
Corneal Tomography

Abstract Purpose The aim of this study is to compare the long-term effects of transepithelial corneal crosslinking with two continuous cycles of iontophoresis (EI-CXL) and conventional corneal crosslinking (C-CXL) in adults with progressive keratoconus. Methods A retrospective analysis was conducted in adults who underwent C-CXL or EI-CXL between 2013 and 2015. Visual acuity, corneal tomography, anterior segment optical coherence tomography, in vivo corneal confocal microscopy (IVCM), and endothelial cell count (ECC) were performed preoperatively and 5 years postoperatively. Results Sixty-eight patients with a mean age of (24.3 ± 3.8) years were included, 34 for each group. After CXL, UCVA or BCVA remained stable, while the spherical diopter, cylinder diopter, spherical equivalent, and Kmax significantly decreased at 1, 2, and 3 years in both groups than baseline (P < 0.05). No significant differences were found in any refractive or tomographic parameters as well as the minimal corneal thickness between groups during follow-up. At 5 years, Kmax was slightly higher in EI-CXL group (58.16 ± 6.28) than that of C-CXL group (57.46 ± 4.98). At 3 and 5 years, the minimal corneal thickness in C-CXL group was still significantly lower than baseline (P < 0.05). IVCM demonstrated the demarcation zone at a mean depth of (302.0 ± 41.7) μm after C-CXL, and at (251.2 ± 28.1) μm after EI-CXL (P < 0.001). Keratocyte repopulation was detectable at all follow-up timepoint in both groups. Postoperative complications including progression were recorded in 6 patients (11.7%) after C-CXL and 3 patients (8.8%) after EI-CXL. ECC remained stable in both groups. Conclusion EI-CXL showed approximate efficacy with C-CXL in stabilizing progressive keratoconus in adults. EI-CXL has the potential to be a preferable transepithelial protocol.

Aptamer Mediated Inhibition of Protein S

Blood ◽  
2016 ◽  
Vol 128 (22) ◽  
pp. 4946-4946
Author(s):  
William E. Plautz ◽  
Juliana Layzer ◽  
Bruce A. Sullenger ◽  
Rinku Majumder
Keyword(s):  
Conflicts Of Interest ◽  
Ex Vivo ◽  
Coagulation Factor ◽  
Protein S ◽  
Factor Ix ◽  
Hemophilia B ◽  
Factor X ◽  
Initial State ◽  
Gla Domain

Abstract Protein S (PS) is an anticoagulant that acts as a direct allosteric inhibitor of the activated form of factor IX (FIXa); FIXa is a coagulant, in both the presence and absence of its cofactor, factor VIIIa. Inhibition of FIXa by PS results in a retarded rate of activation of factor X; FX plays an important role as the initial protease common to the two pathways of blood coagulation. Factor IX (FIX) deficiencies present in the form of varying severities of hemophilia B; additionally, elevated levels of PS result in lengthened clotting times. We intend to increase the efficacy and functional lifetime of natural and infused FIXa in hemophilia B patients by creating a high affinity aptamer that selectively inhibits PS in its interaction with FIXa. Although inhibitory aptamers that target pro-coagulant proteins have been produced in a facile manner, the hemostasis field has yet to direct its sights towards a therapeutically viable aptamer that inhibits a naturally occurring anticoagulant. Through the use of previously selected, and experimentally successful, aptamer pools targeting either Gla-domain or plasma proteomes, we were able to use SELEX to select novel PS ligands. After two rounds of aptamer selection, using the focused pools as an initial state, we have found by nitrocellulose binding assays a strong enrichment from the focused Gla domain proteome, whereas the plasma proteome showed a weaker response. Following development of an inhibitory PS aptamer, we will perform fluorescence binding and substrate cleavage studies to obtain kinetic constants, and ex vivo studies, including aPTT and thrombin generation assays, to demonstrate aptamer efficacy in a physiological system. Ultimately, we plan to test the aptamer in vivo, to demonstrate its use as a possible adjunct therapy to current treatments of hemophilia B. Disclosures No relevant conflicts of interest to declare.

The Role of Polyphenols in the Modulation of Plasma Non-Enzymatic Antioxidant Capacity (NEAC)

2012 ◽  
Vol 82 (3) ◽  
pp. 228-232 ◽  
Author(s):  
Mauro Serafini ◽  
Giuseppa Morabito
Keyword(s):  
Antioxidant Capacity ◽  
Dietary Intervention ◽  
Ex Vivo ◽  
The Body ◽  
Dietary Polyphenols ◽  
Enzymatic Antioxidant ◽  
Endogenous Antioxidant

Dietary polyphenols have been shown to scavenge free radicals, modulating cellular redox transcription factors in different in vitro and ex vivo models. Dietary intervention studies have shown that consumption of plant foods modulates plasma Non-Enzymatic Antioxidant Capacity (NEAC), a biomarker of the endogenous antioxidant network, in human subjects. However, the identification of the molecules responsible for this effect are yet to be obtained and evidences of an antioxidant in vivo action of polyphenols are conflicting. There is a clear discrepancy between polyphenols (PP) concentration in body fluids and the extent of increase of plasma NEAC. The low degree of absorption and the extensive metabolism of PP within the body have raised questions about their contribution to the endogenous antioxidant network. This work will discuss the role of polyphenols from galenic preparation, food extracts, and selected dietary sources as modulators of plasma NEAC in humans.

Antithrombotic Effects and Bleeding Time Prolongation with Synthetic Platelet GPIIb/llla Inhibitors in Animal Models of Platelet-Mediated Thrombosis

1994 ◽  
Vol 71 (01) ◽  
pp. 095-102 ◽  
Author(s):  
Désiré Collen ◽  
Hua Rong Lu ◽  
Jean-Marie Stassen ◽  
Ingrid Vreys ◽  
Tsunehiro Yasuda ◽  
...  
Keyword(s):  
Platelet Aggregation ◽  
Bleeding Time ◽  
Bolus Injection ◽  
Cell Injury ◽  
Ex Vivo ◽  
Thrombus Formation ◽  
Femoral Vein ◽  
Thrombotic Occlusion ◽  
Antithrombotic Effects

SummaryCyclic Arg-Gly-Asp (RGD) containing synthetic peptides such as L-cysteine, N-(mercaptoacetyl)-D-tyrosyl-L-arginylglycyl-L-a-aspartyl-cyclic (1→5)-sulfide, 5-oxide (G4120) and acetyl-L-cysteinyl-L-asparaginyl-L-prolyl-L-arginyl-glycyl-L-α-aspartyl-[0-methyltyrosyl]-L-arginyl-L-cysteinamide, cyclic 1→9-sulfide (TP9201) bind with high affinity to the platelet GPIIb/IIIa receptor.The relationship between antithrombotic effect, ex vivo platelet aggregation and bleeding time prolongation with both agents was studied in hamsters with a standardized femoral vein endothelial cell injury predisposing to platelet-rich mural thrombosis, and in dogs with a carotid arterial eversion graft inserted in the femoral artery. Intravenous administration of G4120 in hamsters inhibited in vivo thrombus formation with a 50% inhibitory bolus dose (ID50) of approximately 20 μg/kg, ex vivo ADP-induccd platelet aggregation with ID50 of 10 μg/kg, and bolus injection of 1 mg/kg prolonged the bleeding time from 38 ± 9 to 1,100 ± 330 s. Administration of TP9201 in hamsters inhibited in vivo thrombus formation with ID50 of 30 μg/kg, ex vivo platelet aggregation with an ID50 of 50 μg/kg and bolus injection of 1 mg/kg did not prolong the template bleeding time. In the dog eversion graft model, infusion of 100 μg/kg of G4120 over 60 min did not fully inhibit platelet-mediated thrombotic occlusion but was associated with inhibition of ADP-induccd ex vivo platelet aggregation and with prolongation of the template bleeding time from 1.3 ± 0.4 to 12 ± 2 min. Infusion of 300 μg/kg of TP9201 over 60 min completely prevented thrombotic occlusion, inhibited ex vivo platelet aggregation, but was not associated with prolongation of the template bleeding time.TP9201, unlike G4120, inhibits in vivo platelet-mediated thrombus formation without associated prolongation of the template bleeding time.

Significance of Platelet β-Adrenoceptors for Platelet Responses In Vivo and In Vitro

1992 ◽  
Vol 68 (06) ◽  
pp. 687-693 ◽  
Author(s):  
P T Larsson ◽  
N H Wallén ◽  
A Martinsson ◽  
N Egberg ◽  
P Hjemdahl
Keyword(s):  
Ex Vivo ◽  
High Dose ◽  
Platelet Aggregability ◽  
Adrenoceptor Agonist ◽  
Dose Infusion ◽  
Von Willebrand ◽  
Willebrand Factor ◽  
Factor Antigen

SummaryThe significance of platelet β-adrenoceptors for platelet responses to adrenergic stimuli in vivo and in vitro was studied in healthy volunteers. Low dose infusion of the β-adrenoceptor agonist isoprenaline decreased platelet aggregability in vivo as measured by ex vivo filtragometry. Infusion of adrenaline, a mixed α- and β-adrenoceptor agonist, increased platelet aggregability in vivo markedly, as measured by ex vivo filtragometry and plasma β-thromboglobulin levels. Adrenaline levels were 3–4 nM in venous plasma during infusion. Both adrenaline and high dose isoprenaline elevated plasma von Willebrand factor antigen levels β-Blockade by propranolol did not alter our measures of platelet aggregability at rest or during adrenaline infusions, but inhibited adrenaline-induced increases in vWf:ag. In a model using filtragometry to assess platelet aggregability in whole blood in vitro, propranolol enhanced the proaggregatory actions of 5 nM, but not of 10 nM adrenaline. The present data suggest that β-adrenoceptor stimulation can inhibit platelet function in vivo but that effects of adrenaline at high physiological concentrations are dominated by an α-adrenoceptor mediated proaggregatory action.

In Vivo Effect of Suloctidil as an Antiplatelet Agent

1979 ◽  
Vol 41 (03) ◽  
pp. 465-474 ◽  
Author(s):  
Marcia R Stelzer ◽  
Thomas S Burns ◽  
Robert N Saunders
Keyword(s):  
Ex Vivo ◽  
Antiplatelet Agent ◽  
Ratio Method ◽  
Platelet Aggregate ◽  
Permanent Effect ◽  
Platelet Aggregates ◽  
5 Ht Uptake ◽  
High Doses

SummaryThe relationship between the effects of suloctidil in vivo as an antiplatelet agent and in vitro as a modifier of platelet serotonin (5-HT) parameters was investigated. Suloctidil was found to be effective in reducing platelet aggregates formation in the retired breeder rat as determined using the platelet aggregate ratio method (PAR) with an ED50 of 16.1 mg/kg 24 hours post administration. In contrast to the hypothesis that 5-HT depletion is involved in the anti-aggregatory mechanism of suloctidil, no correlation was found between platelet 5- HT content and this antiplatelet activity. Reduction of platelet 5-HT content required multiple injections of high doses (100 mg/kg/day) of suloctidil. Suloctidil administration for 8 days at 100 mg/kg/day, which lowered platelet 5-HT content by 50%, resulted in no permanent effect on ex vivo platelet 5-HT uptake or thrombin-induced release, nor alteration in the plasma 5-HT level. However, these platelets exhibited a short-lived, significant increase in percent leakage of 5-HT after 30 minutes of incubation. Therefore, suloctidil treatment at high doses may with time result in platelet 5-HT depletion, however this effect is probably not related to the primary anti-aggregatory activity of the drug.

Studies on the Haemostatic Defect in a Complicated Syndrome

1995 ◽  
Vol 74 (05) ◽  
pp. 1244-1251 ◽  
Author(s):  
H Stormorken ◽  
H Holmsen ◽  
R Sund ◽  
K S Sakariassen ◽  
T Hovig ◽  
...  
Keyword(s):  
Ex Vivo ◽  
Bleeding Tendency ◽  
Clot Retraction ◽  
Abnormal Finding ◽  
Shear Rates ◽  
Perfusion Chamber ◽  
Coagulant Activity ◽  
5 Ht Uptake ◽  
And Storage

SummaryThe Stormorken syndrome is a multifacetted syndrome including a bleeding tendency. No deviations were found in the coagulation- or fibrinolytic systems. Platelet number was low normal, and size abnormal, whereas EM findings were unremarkable. Survival time was half normal. Clot retraction was initially rapid, but clearly decreased, whereas prothrombin consumption was also initially rapid, but complete. Membrane GP’s were normal, so was AA metabolism, PI-cycle, granule storage and secretion, and c-AMP function, whereas 5-HT uptake and storage was decreased. Optical platelet aggregation was low normal with all physiological agonists. The only clearly abnormal finding was that coagulant activity was present on non stimulated platelets at the same level as kaolin-stimulated normal platelets. This indicated a platelet abnormality which should lead to a thrombogenic, not to a haemorrhagic trait. This paradox may have its origin in rheology, because when challenged with in vivo shear rates in an ex vivo perfusion chamber, platelet cohesion was abnormally low. Further studies to better delineate the membrane abnormality are underway.

A Comparative Ex Vivo Study of Plasminogen Activators and Proteases for Fibrinolytic Activity and Side Effects in Rabbits

1977 ◽  
Vol 37 (01) ◽  
pp. 154-161 ◽  
Author(s):  
B. A Janik ◽  
S. E Papaioannou
Keyword(s):  
Side Effects ◽  
Effective Dose ◽  
Fibrinolytic Activity ◽  
Ex Vivo ◽  
Plasminogen Activators ◽  
Assay System ◽  
Coagulation Parameters ◽  
Ex Vivo Assay

SummaryUrokinase, streptokinase, Brinase, trypsin, and SN 687, a bacterial exoprotease, have been evaluated in an ex vivo assay system. These enzymes were injected into rabbits and the fibrinolytic activity as well as other coagulation parameters were measured by in vitro techniques. Dose-response correlations have been made using the euglobulin lysis time as a measure of fibrinolytic activity and the 50% effective dose has been determined for each enzyme. Loading doses, equal to four times the 50% effective dose, were administered to monitor potential toxicity revealing that Brinase, trypsin, and SN 687 were very toxic at this concentration.Having established the 50% effective dose for each enzyme, further testing was conducted where relevant fibrinolytic and coagulation parameters were measured for up to two days following a 50% effective dose bolus injection of each enzyme. Our results have demonstrated that urokinase and streptokinase are plasminogen activators specifically activating the rabbit fibrinolytic system while Brinase, trypsin and SN 687 increase the general proteolytic activity in vivo.The advantages of this ex vivo assay system for evaluating relative fibrinolytic potencies and side effects for plasminogen activators and fibrinolytic proteases have been discussed.

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