A Novel Method for the Rapid Purification of Human and Rat Fibrin(OGEN) Degradation Products in High Yields

1979 ◽  
Author(s):  
W. Nieuwenhuizen ◽  
I. A. M. van Ruijven-Vermeer ◽  
F. Haverkate ◽  
G. Timan

A novel method will be described for the preparation and purification of fibrin(ogen) degradation products in high yields. The high yields are due to two factors. on the one hand an improved preparation method in which the size heterogeneity of the degradation products D is strongly reduced by plasmin digestion at well-controlled calcium concentrations. At calcium concentrations of 2mM exclusively D fragments, M.W.= 93-000 (Dcate) were formed; in the presence of 1OmM EGTA only fragments M.W.= 80.000 (D EGTA) were formed as described. on the other hand a new purification method, which includes Sephadex G-200 filtration to purify the D:E complexes and separation of the D and E fragments by a 16 hrs. preparative isoelectric focussing. The latter step gives a complete separation of D (fragments) (pH = 6.5) and E fragments (at pH = 4.5) without any overlap, thus allowing a nearly 100% recovery in this step. The overall recoveries are around 75% of the theoretical values. These recoveries are superior to those of existing procedures. Moreover the conditions of this purification procedure are very mild and probably do not affect the native configuration of the products. Amino-terminal amino acids of human Dcate, D EGTA and D-dimer are identical i.e. val, asx and ser. in the ratgly, asx and ser were found. E 1% for rat Dcate=17-8 for rat D EGTA=16.2 and for rat D- dimer=l8.3. for the corresponding human fragments, these values were all 20.0 ± 0.2.

1979 ◽  
Author(s):  
W. Nieuwenhuizen ◽  
I.A.M. van Ruijven-Vermeer ◽  
F. Haverkate ◽  
G. Timan

A novel method will be described for the preparation and purification of fibrin(ogen) degradation products in high yields. The high yields are due to two factors. On the one hand an improved preparation method in which the size heterogeneity of the degradation products D is strongly reduced by plasmin digestion at well-control 1ed calcium concentrations. At calcium concentrations of 2mM exclusively D fragments, M.W.= 93.000 (Dcate) were formed; in the presence of 10mM EGTA only fragments M.W.= 80.000 (D EGTA) were formed as described. On the other hand a new purification method, which includes Sephadex G-200 filtration to purify the D:E complexes and separation of the D and E fragments by a 16 hrs. preparative isoelectric focussing. The latter step gives a complete separation of D (fragments) (pH = 6.5) and E fragments (at pH = 4.5) without any overlap, thus allowing a nearly 100% recovery in this step. The overall recoveries are around 75% of the theoretical values. These recoveries are superior to those of existing procedures. Moreover the conditions of this purification procedure are very mild and probably do not affect the native configuration of the products. Amino-terminal amino acids of human Dcate, D EGTA and D-dimer are identical i.e. val, asx and ser. In the ratgly, asx and ser were found. E 1% for rat Dcate=17.8 for rat D EGTA=16.2 and for rat D-dimer=l8.3. For the corresponding human fragments, these values were all 20.0 ± 0.2.


1979 ◽  
Vol 360 (1) ◽  
pp. 633-638 ◽  
Author(s):  
Irina A. M. VAN RUIJVEN-VERMEER ◽  
Willem NIEUWENHUIZEN ◽  
Frits HAVERKATE ◽  
Truus TIMAN

Author(s):  
Ariaki Nagayama

Vinblastine(Vb) or vincristine, alkaloid derived from Vinca rosea is known for its antimitotic activity by regrouping of microtubules into paracrystalline form within the cells. A rapid purification method of vinblastine-induced microtubular paracrystals(PC) has provided us with a fresh and pure microtubular material demonstrating the presence of a labile ATPase associated with the PC. The present report is concerned with the fine structure of purified microtubules of mammalian cultured cells.Confluent monolayer cultures of L cells were incubated for 20hrs with 10-5 M Vb (donated from Shionogi Seiyaku & Co., Osaka, Japan).


2012 ◽  
Vol 58 (2) ◽  
pp. 177-192 ◽  
Author(s):  
Marek Parfieniuk ◽  
Alexander Petrovsky

Near-Perfect Reconstruction Oversampled Nonuniform Cosine-Modulated Filter Banks Based on Frequency Warping and Subband MergingA novel method for designing near-perfect reconstruction oversampled nonuniform cosine-modulated filter banks is proposed, which combines frequency warping and subband merging, and thus offers more flexibility than known techniques. On the one hand, desirable frequency partitionings can be better approximated. On the other hand, at the price of only a small loss in partitioning accuracy, both warping strength and number of channels before merging can be adjusted so as to minimize the computational complexity of a system. In particular, the coefficient of the function behind warping can be constrained to be a negative integer power of two, so that multiplications related to allpass filtering can be replaced with more efficient binary shifts. The main idea is accompanied by some contributions to the theory of warped filter banks. Namely, group delay equalization is thoroughly investigated, and it is shown how to avoid significant aliasing by channel oversampling. Our research revolves around filter banks for perceptual processing of sound, which are required to approximate the psychoacoustic scales well and need not guarantee perfect reconstruction.


Author(s):  
J Ph Guillet ◽  
E Pilon ◽  
Y Shimizu ◽  
M S Zidi

Abstract This article is the first of a series of three presenting an alternative method of computing the one-loop scalar integrals. This novel method enjoys a couple of interesting features as compared with the method closely following ’t Hooft and Veltman adopted previously. It directly proceeds in terms of the quantities driving algebraic reduction methods. It applies to the three-point functions and, in a similar way, to the four-point functions. It also extends to complex masses without much complication. Lastly, it extends to kinematics more general than that of the physical, e.g., collider processes relevant at one loop. This last feature may be useful when considering the application of this method beyond one loop using generalized one-loop integrals as building blocks.


2021 ◽  
Vol 76 (2) ◽  
pp. 85-90
Author(s):  
Abdolkarim Zare ◽  
Manije Dianat

Abstract A highly efficient and green protocol for the synthesis of pyrimido[4,5-b]quinolines has been described. The one-pot multicomponent reaction of dimedone with arylaldehydes and 6-amino-1,3-dimethyluracil in the presence of N,N-diethyl-N-sulfoethanaminium chloride ([Et3N–SO3H][Cl]) as an ionic liquid (IL) catalyst under solvent-free conditions afforded the mentioned compounds in high yields and short reaction times. Our protocol is superior to many of the reported protocols in terms of two or more of these factors: the reaction times, yields, conditions (solvent-free versus usage of organic solvents), temperature and catalyst amount.


FEBS Letters ◽  
1978 ◽  
Vol 93 (2) ◽  
pp. 361-364 ◽  
Author(s):  
Gert Pflugfelder ◽  
Johann Sonnenbichler

1987 ◽  
Author(s):  
P J Gaffney ◽  
L J Creighton ◽  
A Curry ◽  
B MacMahon ◽  
R Thorpe

Monoclonal antibodies (mabs) to crosslinked fibrin degradation products (XL-FDP) having the general formula D/Y[X]nY/D (known as X-oligomer) and D-D (known as D dimer) have been raised in balb/C mice by both a novel mtrasplenic and a conventional subcutaneous route of immunisation and by combinations of both these procedures. Mabs to X-oligomers (NIBn 52 and NIBn 123) obtained by an intrasplenic procedure have been demonstrated to crossreact only with X-oligomer in a 2-site ELISA procedure and not with D dimer or whole fibrinogen and have been shown to be of value m the examination of clinical material obtained from patients with various types of thrombosis and have also been useful in monitoring the efficacy of thrombolytic therapy. The X-oligomer mabs are immunoglobulins of the M class. It was demonstrated that their unique specificity for conformational epitopes on the large X-oligomer fragments does not reside in the IgM structure since alterative immunisation procedures have been used to generate mabs of the IgG class which have the same specificity. Using immunoglobulin class switching in culture rather than during immunisation was suggested by certain cell lines which produced both IgM and IgG specific for X-oligomer. This latter point needs rigorous validation.Immunoglobulin G type mabs to highly purified D dimer were raised by conventional subcutaneous immunisation of balb/C mice. One of these, NIBn-11, was found to crossreact with PVC-immobilised X-oligomer and D dimer but not with fibrinogen. However NIBn-11 did not bind to D dimer in a 2-site ELISA procedure while crossreactmg quite avidly with X-oligomer. This suggests that the D dimer epitope to which NIBn-11 is directed is expressed in some conformations and not m others and that these conformations are always expressed in the complex X-oligomer group of fragments. These mabs, whilst of value in measuring certain unique fibrin fragments m plasma, are useful in the epitope mapping of fibrinogen/fibrin and their plasmm-mediated


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