scholarly journals CHEMICAL, CLINICAL AND IMMUNOLOGICAL STUDIES ON THE PRODUCTS OF HUMAN PLASMA FRACTIONATION. XXXIII. THE COAGULATION DEFECT IN HEMOPHILIA: THE EFFECT IN VITRO AND IN VIVO ON THE COAGULATION TIME IN HEMOPHILIA OF A PROTHROMBIN AND FIBRINOGEN-FREE NORMAL PLASMA AND ITS DERIVED PROTEIN FRACTIONS 123

1946 ◽  
Vol 25 (6) ◽  
pp. 876-879 ◽  
Author(s):  
Jessica H. Lewis ◽  
J. P. Soulier ◽  
F. H. L. Taylor
Keyword(s):  
Human Plasma ◽  
Protein Fractions ◽  
Coagulation Time ◽  
Coagulation Defect ◽  
Normal Plasma

COMPARISON OF THE EFFECT OF FACTOR VII PREPARED FROM HUMAN PLASMA (pVIIa) AND RECOMBINANT Vila (rVIIa) IN VITRO AND IN RABBITS

1987 ◽  
Author(s):  
U Hender ◽  
T Lund-Hansen ◽  
D Winther
Keyword(s):  
Human Plasma ◽  
Factor Vii ◽  
Normal Plasma ◽  
Vitro Effect ◽  
Higher Doses

Purified human FVIIa has been shown to induce haemostasis in pat. with haemoph. A compl. with antibodies against VIII:C (Hedner & Kisiel 1983). The in vitro effect of addition of pVIIa or rVIIa was studied by adding 0, 44, or 180 u pVIIa/rVIIa/ml to haemoph. A or haemoph. B plasma. The APTT shortened from 61 s (mean of 5 determin.) without any Vila added to 38 s after addition of 44 u pVIIa to haemoph. A plasma, and to 33 s after addition of 180 u/ml. In haemoph. B plasma the corresponding APTTs were 64 s, 35 s (44 u pVIIa/ml Vila) and 31 s (180 u pVIIa/ ml). Similar results were obtained when rVIIa was added to the same plasmas. In haemoph. A plasma APTT was shortened from 66 S to 52 s (50 u rVIIa/ml) and to41s(159u rVIIa/ml), and in haemoph. B plasma 71 s,44 s,and 37 s.Also in plasma from pat. with acquired antibodies against VIII:C a shortening of APTT was found both of pVIIa (60 s, 37 s, and 32 s) and rVIIs (65 s, 38 s, and 33 s). In normal plasma the APTT only shortened a few seconds after addition of the same amounts of pVIIa/rVIIa per ml (30 s, 27 s, 25 s). The doses required to normalize the APTT in vitro exceed substantially the doses used in vivo so far. The pVIIa and rVIIa were therefore also given i.v. to rabbits.Plasma samples were drawn before inj., 15 min, 2 hrs,4 hrs, 8 hrs, and 24 hrs after and platelets, APTT, fib.gen, ATIII, α2M, α2AP, ethanol gel. test, FVII,Hb, Hkr were followed. Doses of 500 u/kgb.w.,1000 or 5000 u/kg b.w. were given. No signs of a gen.activ.of the coag. were observed. The FVII level in plasm rose adequately. In conclusion higher doses of Vila than used clin. so far may be needed to achieve full haemost. effect in haemoph. pat. or pat. with acq. antibodies against VIII:C. Such doses also seem to be safe. rVIIa was as active as pVIIa and as safe in rabbits.

Potentially Thrombogenic Materials in Factor IX Concentrates

1975 ◽  
Vol 33 (03) ◽  
pp. 617-631 ◽  
Author(s):  
H. S Kingdon ◽  
R. L Lundblad ◽  
J. J Veltkamp ◽  
D. L Aronson
Keyword(s):  
Human Plasma ◽  
Antithrombin Iii ◽  
Factor Ix ◽  
In Vitro Assays ◽  
Substantial Agreement ◽  
Coefficient Of Correlation

SummaryFactor IX concentrates manufactured from human plasma and intended for therapeutic infusion in man have been suspected for some time of being potentially thrombogenic. In the current studies, assays were carried out in vitro and in vivo for potentially thrombogenic materials. It was possible to rank the various materials tested according to the amount of thrombogenic material detected. For concentrates not containing heparin, there was substantial agreement between the in vivo and in vitro assays, with a coefficient of correlation of 0.77. There was no correlation between the assays for thrombogenicity and the antithrombin III content. We conclude that many presently available concentrates of Factor IX contain substantial amounts of potentially thrombogenic enzymes, and that this fact must be considered in arriving at the decision whether or not to use them therapeutically.

Presence and Possible Origin of ɛ(γ-Glutamyl)Lysine Isodipeptide in Human Plasma

1992 ◽  
Vol 67 (01) ◽  
pp. 060-062 ◽  
Author(s):  
J Harsfalvi ◽  
E Tarcsa ◽  
M Udvardy ◽  
G Zajka ◽  
T Szarvas ◽  
...  
Keyword(s):  
Human Plasma ◽  
Fibrinolytic Therapy ◽  
Normal Human Plasma ◽  
Normal Human ◽  
Hplc Technique ◽  
Significant Change

Summaryɛ(γ-glutamyl)lysine isodipeptide has been detected in normal human plasma by a sensitive HPLC technique in a concentration of 1.9-3.6 μmol/1. Incubation of in vitro clotted plasma at 37° C for 12 h resulted in an increased amount of isodipeptide, and there was no further significant change when streptokinase was also present. Increased in vivo isodipeptide concentrations were also observed in hypercoagulable states and during fibrinolytic therapy.

scholarly journals Ketorolac-dextran conjugates: Synthesis, in vitro and in vivo evaluation

2007 ◽  
Vol 57 (4) ◽  
pp. 441-450 ◽  
Author(s):  
Savita Vyas ◽  
Piyush Trivedi ◽  
Subhash Chaturvedi
Keyword(s):  
Human Plasma ◽  
Parent Drug ◽  
Aqueous Solubility ◽  
Spectrophotometric Analysis ◽  
In Vivo Evaluation ◽  
Gastrointestinal Side Effects ◽  
Molecular Masses ◽  
Anti Inflammatory

Ketorolac-dextran conjugates: Synthesis,in vitroandin vivoevaluationKetorolac is a non-steroidal anti-inflammatory drug. Dextran conjugates of ketorolac (KD) were synthesized and characterized to improve ketorolac aqueous solubility and reduce gastrointestinal side effects. An N-acylimidazole derivative of ketorolac (KAI) was condensed with a model carrier polymer, dextran of different molecular masses (40000, 60000, 110000 and 200000). IR spectral data confirmed formation of ester bonding. Ketorolac contents were evaluated by UV-spectrophotometric analysis. The molecular mass was determined by measuring viscosity using the Mark-Howink-Sakurada equation. Invitrohydrolysis studies were performed in aqueous buffers (pH 1.2, 7.4, 9) and in 80% (V/V) human plasma (pH 7.4). At pH 9, a higher rate of ketorolac release from KD was observed as compared to aqueous buffer of pH 7.4 and 80% human plasma (pH 7.4), following first-order kinetics.In vivobiological screening in mice and rats indicated that conjugates retained analgesic and anti-inflammatory activities with significantly reduced ulcerogenicity compared to the parent drug.

scholarly journals ELISA detection of MPO-DNA complexes in human plasma is error-prone and yields limited information on neutrophil extracellular traps formed in vivo

PLoS ONE ◽  
2021 ◽  
Vol 16 (4) ◽  
pp. e0250265
Author(s):  
Hubert Hayden ◽  
Nahla Ibrahim ◽  
Johannes Klopf ◽  
Branislav Zagrapan ◽  
Lisa-Marie Mauracher ◽  
...  
Keyword(s):  
Human Plasma ◽  
Dynamic Range ◽  
Neutrophil Extracellular Traps ◽  
Plasma Samples ◽  
Dna Complexes ◽  
High Background ◽  
Isotype Control ◽  
Extracellular Traps

Over the past years, neutrophil extracellular traps (NETs) were shown to contribute to states of acute and chronic inflammatory disease. They are composed of expelled chromatin and decorated by neutrophil-derived proteins. Therefore, the analysis of DNA complexes with myeloperoxidase (MPO) by ELISA has become an attractive tool to measure NET formation in in vitro and in vivo samples. When we used a published MPO-DNA ELISA protocol and included an isotype control for the anti-MPO coating antibody, we observed high assay specificity for in vitro prepared NET samples, whereas the specificity for in vivo plasma samples was low. In addition, the assay failed to detect in vitro generated MPO-DNA complexes when spiked into plasma. Therefore, we set out to improve the specificity of the MPO-DNA ELISA for plasma samples. We found that the use of Fab fragments or immunoglobulins from different species or reversal of the antibody pair led to either a high background or a low dynamic range of detection that did not improve the specificity for plasma samples. Also, the use of higher plasma dilutions or pre-clearing of plasma immunoglobulins were ineffective. Finally, we found that a commercial reagent designed to block human anti-mouse antibodies and multivalent substances increased the detection window between the MPO antibody and isotype control for highly diluted plasma. We applied this modified ELISA protocol to analyze MPO-DNA complexes in human blood samples of acute and chronic inflammatory conditions. While markers of neutrophil activation and NET formation such as MPO, elastase and citrullinated histone H3 correlated significantly, we observed no correlation with the levels of MPO-DNA complexes. Therefore, we conclude that ELISA measurements of MPO-DNA complexes in human plasma are highly questionable regarding specificity of NET detection. In general, plasma analyses by ELISA should more frequently include isotype controls for antibodies to demonstrate target specificity.

scholarly journals Gene expression–based discovery of atovaquone as a STAT3 inhibitor and anticancer agent

Blood ◽  
2016 ◽  
Vol 128 (14) ◽  
pp. 1845-1853 ◽  
Author(s):  
Michael Xiang ◽  
Haesook Kim ◽  
Vincent T. Ho ◽  
Sarah R. Walker ◽  
Michal Bar-Natan ◽  
...  
Keyword(s):  
Gene Expression ◽  
Retrospective Study ◽  
Human Plasma ◽  
Patient Outcomes ◽  
Anticancer Agent ◽  
Plasma Concentrations ◽  
Anticancer Efficacy ◽  
Key Points

Key PointsThe FDA-approved drug atovaquone is a novel, clinically available inhibitor of STAT3 at standard human plasma concentrations. Atovaquone shows anticancer efficacy in vitro, in vivo, and in a retrospective study of AML patient outcomes after atovaquone treatment.

scholarly journals Activation of human factor VII during clotting in vitro

Blood ◽  
1985 ◽  
Vol 65 (1) ◽  
pp. 218-226 ◽  
Author(s):  
LV Rao ◽  
SP Bajaj ◽  
SI Rapaport
Keyword(s):  
Tissue Factor ◽  
Glass Tube ◽  
Factor Vii ◽  
Clotting Factor ◽  
Activate Factor ◽  
Factor Xii ◽  
Factor X ◽  
Normal Plasma

Abstract We have studied factor VII activation by measuring the ratio of factor VII clotting to coupled amidolytic activity (VIIc/VIIam) and cleavage of 125I-factor VII. In purified systems, a low concentration of Xa or a higher concentration of IXa rapidly activated 125I-factor VII, yielding a VIIc/VIIam ratio of 25 and similar gel profiles of heavy and light chain peaks of VIIa. On further incubation, VIIa activity diminished and a third 125I-peak appeared. When normal blood containing added 125I- factor VII was clotted in a glass tube, the VIIc/VIIam ratio rose fivefold, and 20% of the 125I-factor VII was cleaved. Clotting normal plasma in an activated partial thromboplastin time (APTT) system yielded a VIIc/VIIam ratio of 25 and over 90% cleavage of 125I-factor VII. Clotting factor XII-deficient plasma preincubated with antibodies to factor X in an APTT system with added XIa yielded a VIIc/VIIam ratio of 19 and about 60% cleavage, which indicates that IXa, at a concentration achievable in plasma, can effectively activate factor VII. Clotting normal plasma with undiluted tissue factor yielded a VIIc/VIIam ratio of 15 to 20 and 60% cleavage of 125I-factor VII, whereas clotting plasma with diluted tissue factor activated factor VII only minimally. We conclude that both Xa and IXa can function as significant activators of factor VII in in vitro clotting mixtures but believe that only small amounts of factor VII may be activated in vivo during hemostasis.

DETECTION OF INACTIVATED α2-MACROGLOBULIN (α2M) IN HUMAN PLASMA USING MONOCLONAL ANTIBODIES

1987 ◽  
Author(s):  
J Abbink ◽  
J Nuijens ◽  
C Huijbregts ◽  
E Hack
Keyword(s):  
Monoclonal Antibodies ◽  
Longitudinal Studies ◽  
Human Plasma ◽  
Dextran Sulfate ◽  
Optimal Conditions ◽  
Α2 Macroglobulin ◽  
In Vitro Activation

Monoclonal antibodies (mAbs) were raised against human a2M. Five mAbs that bound to α2M in ELISA were further analyzed by a radioimmunoassay (RIA) for their reaction with three types of α2M: native α2M, chemically inactivated α2M (iα2M) (methylamine treated), and proteolytically iα2M. One mAb reacted with all forms of α2M, while four mAbs bound both forms of ia2M but not native α2M. One of these latter mAbs (Ml) was used to develop a RIA (the Ml-assay) for the detection of iα2M in plasma: Ml coupled to Sepharose is incubated with the plasma to be tested, and bound iα2M is detected by a subsequent incubation with polyclonal 125I-anti-α2M antibodies. As little as 5 ng of iα2M can be detected with this assay in the presence of an excess of native α2M. This assay was then applied to measure inactivation of α2M in vitro and in vivo. In vitro activation of the contact system in plasma by dextran sulfate results in the inactivation of ca 10% of α2M. When blood from normal donors was collected under optimal conditions, about 0.5% of the total α2M content appeared to be iα2M. Longitudinal studies in patients (a.o. with septicaemie, during cardiopulmunary bypass) revealed that increased levels of iα2M occurred sporadically. The Ml-assay appears to be useful to monitor the role of α2M in human diseases.

scholarly journals Plasma Transfusions in Hemophilia

Blood ◽  
1952 ◽  
Vol 7 (7) ◽  
pp. 710-720 ◽  
Author(s):  
S. VAN CREVELD ◽  
M. M. P. PAULSSEN
Keyword(s):  
Two Stage ◽  
Coagulation Time ◽  
Prothrombin Activity ◽  
Lasting Effect ◽  
One Stage ◽  
The One ◽  
Rapid Appearance

Abstract Transfusions of heparinized plasma have a greater and more lasting effect on the coagulation time of hemophiliacs than transfusions of citrated plasma. Both in vitro and in vivo, heparinized plasma causes in hemophiliacs a far greater consumption of prothrombin as determined with the two-stage method than citrated plasma. In using the one-stage method no important differences in prothrombin activity are found after transfusions of heparinized and of citrated plasma respectively. This fact was thought to be connected with the more or less rapid appearance of an accelerator. Its a hemophiliac with a circulating anticoagulant, transfusions of heparinized plasma were unable to shorten the coagulation time to any important degree, nor did these transfusions cause an important decrease of serum prothrombin as determined by the two-stage method.

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