A Simple Rapid Method for Isolating Soluble Fibrin Complexes from Fibrinogen by Treatment with Thrombin and t-AMCHA

1979 ◽  
Vol 42 (05) ◽  
pp. 1388-1397
Author(s):  
Shigeru Hayashi ◽  
Kaneo Yamada
Keyword(s):  
Carboxylic Acid ◽  
Rapid Method ◽  
Recovery Rate ◽  
Gel Filtration ◽  
The Other ◽  
Recovery Rates ◽  
Soluble Fibrin ◽  
Other Hand

SummaryUsing treatment with thrombin associated with trans-aminomethylcyclohexane carboxylic acid (t-AMCHA), a simple and rapid method for isolating soluble fibrin complexes (SFC) from fibrinogen in the plasma was developed. By this procedure, the recovery rates of SFC and early FDP (mainly X) increased according to the concentration of t-AMCHA, reaching a maximum at 286 mM t-AMCHA. On the other hand, the recovery rate of fibrinogen remained below 1.5% and that of late FDP was almost 100% at all concentrations of t- AMCHA. These results suggested that SFC and FDP could be isolated from fibrinogen by thrombin and t-AMCHA (286 mM) treatment. Moreover, it was possible to isolate SFC from FDP using gel filtration after treatment with thrombin and t-AMCHA. The SFC could be quantified by assay of the eluted fractions containing SFC by the staphylococcal clumping test.

scholarly journals Embryo recovery from the oviduct in superovulated ewes: a method to improve MOET systems

2008 ◽  
Vol 53 (No. 4) ◽  
pp. 145-151
Author(s):  
J.P. Ramon-Ugalde ◽  
J. Folch ◽  
M.J. Cocero ◽  
R.E. Piña-Aguilar ◽  
J.L. Alabart
Keyword(s):  
Recovery Rate ◽  
Uterine Horn ◽  
Conception Rate ◽  
The Other ◽  
Embryo Recovery ◽  
Intramuscular Injections ◽  
Other Hand

The efficiency of embryo recovery in a superovulatory treatment was studied by perfusing the oviduct or the uterine horn in 3.5 and 7 days after sponge withdrawal, respectively. Eighty-four and seventy ewes of Aragonesa breed were used as embryo donors and receptors, respectively. The donors were distributed in 3 replications. The oestrus was synchronized with the insertion of FGA intravaginal sponges (30 mg) for 13 days. Six intramuscular injections of 18 mg pFSH were applied in decreasing doses at 12 hour intervals starting 48 hours after sponge withdrawal. The recovery rate (RR) (83.2 vs. 75.8%), the viability rate (VR) (73.5 vs. 47.2%) and the number of viable embryos (VE) per donor ewe (5.9 ± 0.79 vs. 3.0 ± 0.37) were higher (<I>P</I> < 0.01) in ewes whose perfusion was done directly in oviducts. The interval from sponge withdrawal to oestrus had no influence on RR in any groups; however it had an effect on the VR in 3.5 and 7 days old embryos. Embryos from ewes showing the oestrus within 20 hours after sponge withdrawal had a higher viability. On the other hand, using morphologic criteria, 26.5% and 52.7% of 3.5 and 7 days old embryos, respectively, were discarded before transfer. The percentage of ewes in each group that became pregnant was similar in both groups. We conclude that in superovulated ewes the embryo recovery rate, viability and number of viable embryos are higher in embryos recovered from the oviduct compared to those recovered from the uterus, without decreasing the conception rate and the viability rate is higher in ewes that show the oestrus 20 hours before sponge withdrawal.

Malate dehydrogenase from thermophilic Humicola lanuginosa and Mucor pusillus: purification and comparative properties of the enzymes with differing thermostabilities

1984 ◽  
Vol 62 (7) ◽  
pp. 559-565 ◽  
Author(s):  
Anupam S. Wali ◽  
Autar K. Mattoo
Keyword(s):  
Malate Dehydrogenase ◽  
Conformational Changes ◽  
Sodium Dodecyl ◽  
Gel Filtration ◽  
The Other ◽  
Heat Inactivation ◽  
Ph Optimum ◽  
Molecular Weights ◽  
Other Hand ◽  
Humicola Lanuginosa

Malate dehydrogenase (L-malate:NAD+ oxidoreductase, EC 1.1.1.37) was purified from the thermophiles Humicola lanuginosa and Mucor pusillus. The H. lanuginosa enzyme was homogeneous on sodium dodecyl sulphate – polyacrylamide gels, while the M. pusillus enzyme was more than 95% pure. The two enzymes appeared to be composed of two subunits of equal size, each of 36 000 daltons (H. lanuginosa) or 33 000 daltons (M. pusillus). The native enzymes revealed molecular weights of 68 000 as determined by gel filtration. The isoelectric points of malate dehydrogenase from H. lanuginosa and M. pusillus were 3.9 and 4.2, respectively. The reduction of oxaloacetate by the H. lanuginosa enzyme was optimum at pH 8.5–9 with apparent Km's of 0.12 mM for oxaloacetate and 0.027 mM for NADH. On the other hand, M. pusillus enzyme snowed a pH optimum of 7.8–8.5 with apparent Km's of 0.075 mM for oxaloacetate and 0.1 mM for NADH. The L-malate oxidation reaction was catalyzed optimally at pH 10 by the H. lanuginosa enzyme with apparent Km's of 5.8 mM for malate and 0.1 mM for NAD, while the M. pusillus enzyme catalyzed it optimally between pH 9.5 and 10 with apparent Km's of 4.44 mM for malate and 0.16 mM for NAD. The optimum temperature for reduction of oxaloacetate was 50 °C for both the enzymes. The H. lanuginosa enzyme was resistant to heat inactivation at 40 °C, but lost 60% of its activity after 15 min at 50 °C. Mucor pusillus enzyme, on the other hand, retained 90% activity at 60 °C after 10 min. The two enzymes were protected from heat inactivation by monovalent cations (viz Na+, K+, and NH4+), as well as citrate, which may possibly involve conformational changes.

scholarly journals Spectroscopy Study of Honey Pineapple Peels Extracted in Different Solvents

2021 ◽  
Vol 3 (1) ◽  
pp. 32-35
Author(s):  
Yehezkiel Steven Kurniawan ◽  
Edi Setiyono ◽  
Marcelinus Alfasisurya Setya Adhiwibawa ◽  
Krisfian Tata Aneka Priyangga ◽  
Leny Yuliati
Keyword(s):  
Carboxylic Acid ◽  
Visible Region ◽  
Water Extract ◽  
Ethanolic Extract ◽  
Acetone Extract ◽  
The Other ◽  
Distilled Water ◽  
Spectroscopy Study ◽  
Absorption Signal ◽  
Other Hand

In the present work, we investigated the extract of honey pineapple peels in distilled water, ethanol, and acetone solvents. The spectroscopy study of each extract was performed using a Fourier transform infrared (FTIR) spectrometer, an ultraviolet-visible (UV-Vis) spectrophotometer, and a spectrofluorometer. The FTIR spectrum of the distilled water extract indicated that the distilled water extract may contain alcohol or carboxylic acid compounds. Meanwhile, the ethanolic extract may contain alcohol or carboxylic acid, or ether compounds. On the other hand, the acetone extract may contain alcohol or ether or aromatic or aliphatic compounds. The UV-Vis spectrum of the honey pineapple peels extracted in the distilled water, ethanol, and acetone showed a broad absorption signal at UV region (< 300 nm), four absorption signals at UV region (232-368 nm), and four absorption signals at UV region (231-368 nm) with a weak absorption signal at the visible region at 559 nm, respectively. The distilled water and acetone extracts gave fluorescence signals, however, the ethanolic extract showed no fluorescence intensity. From the FTIR, UV-Vis, and fluorescence spectra characterization, the extracted natural pigments from the honey pineapple peels in distilled water, ethanol, and acetone solvents were identified. The distilled water extract may contain polar flavonoid or steroid compounds while the ethanolic extract may contain polar carotenoid pigments. On the other hand, the acetone extract may contain carotenoid and chlorophyll pigments as shown by an emission signal at 670 nm.

Comparison of Ethanol and Protamine Tests in Demonstration of Soluble Fibrin and Early Products of Fibrin Degradation

1972 ◽  
Vol 28 (03) ◽  
pp. 342-350 ◽  
Author(s):  
Y. P Konttinen ◽  
L Kemppainen ◽  
O Turunen
Keyword(s):  
Spectrophotometric Method ◽  
Degradation Products ◽  
The Other ◽  
Clot Lysis ◽  
Fibrinogen Degradation Products ◽  
Soluble Fibrin ◽  
Intravascular Coagulation ◽  
Fibrin Monomer ◽  
Other Hand ◽  
Gelation Test

SummaryPerformance and applicability of ethanol-induced gelation and protamine-induced paracoagulation for the demonstration of soluble fibrin monomer complexes and fragment Xo complexes was studied by using 1. fibrin monomer plasma prepared by adding small amounts of thrombin to plasma, 2. clot lysis products, and 3. thrombin -treated mixture of fibrinogen degradation products and plasma. To increase the specificity of the protamine tests only visible fibrin strand formation was recorded as positive. In addition to qualitative tests the amount of paracoagulable material was measured by a spectrophotometric method.The ethanol gelation test proved very simple, reproducible and considerably more sensitive than the protamine tests in demonstrating soluble fibrin monomer complexes, irrespective of whether fibrinogen degradation products were present or not. On the other hand, the protamine tests were clearly superior for demonstration of clot lysis products (fragment Xo complexes). Therefore it seems advisable to perform both types of tests when screening for intravascular coagulation.

Activation of Serum Thrombotic Accelerator (STA) by Digitonin and Saponin

1967 ◽  
Vol 18 (03/04) ◽  
pp. 375-382
Author(s):  
R. M Howell ◽  
G. B. D Scott
Keyword(s):  
Cardiac Glycosides ◽  
Gel Filtration ◽  
Molecular Size ◽  
Membrane Lipid ◽  
Therapeutic Use ◽  
The Other ◽  
Precursor Complex ◽  
Other Hand ◽  
Original Size

SummaryThe steroid saponins digitonin and saponin white, compounds effective in disrupting membrane lipid-protein, have been shown to activate STA precursor.Ultracentrifuge studies showed a radical alteration in the floatation behaviour of the precursor complex on activation by digitonin, indicating an increase in its molecular size.Gel-filtration studies, on the other hand, revealed that activation by digitonin was reversible, since passage down a Sephadex G-200 column not only removed digitonin but abolished all activity, the latter being fully restored by the further addition of digitonin. Furthermore, since the elution positions of inactive precursor and deactivated STA were identical, this de-activation was accompanied by the return of the complex to its original size. The failure of cardiac glycosides, other than digitonin, to activate the precursor may be due to their being more hydrophobic and suggests that hypercoagulability leading to possible thrombosis does not arise from their therapeutic use.

Fibrinogen-Induced Polymerization Via the Process of Methylation

1979 ◽  
Vol 42 (05) ◽  
pp. 1398-1410
Author(s):  
A J Osbahr
Keyword(s):  
Carboxylic Acid ◽  
Thionyl Chloride ◽  
Acid Group ◽  
Fibrin Clot ◽  
Spectrophotometric Analysis ◽  
The Other ◽  
Amino Group ◽  
Methyl Sulfate ◽  
Carboxylic Acid Groups ◽  
Other Hand

SummaryFibrinogen is polymerized by a number of group specific reagents including diazomethane, thionyl chloride and di-methyl sulfate at pH 7.4. The relationship between the number of methyl groups incorporated into fibrinogen and the extent of polymerization was evaluated. With diazomethane and thionyl chloride as modifying agents, polymerization ensued in approximately IV2 hr with extensive modification of fibrinogen. On the other hand, m�thylation via di-methyl sulfate-induced polymer formation occurs in approximately 35 min with primarily carboxylic acid group esterification. The polymerized fibrinogen formed under these conditions exhibited properties that were closely similar with the physiological fibrin clot.Amino group determinations revealed the m�thylation of amino acid residues other than the expected esterification of carboxylic acid groups. Diazomethane induced both N- methylation of lysine, as well as O-methylation of tyrosine, as estimated from spectrophotometric analysis. On the other hand, thionyl chloride modified only a small number of amino groups, and di-methyl sulfate modification resulted in no significant amounts of amino group methylation during the process of modification induced polymerization of fibrinogen.The profile of the number of methoxyl groups incorporated into fibrinogen with time for diazomethane modification may reflect a conformational change in the protein due to a more nonspecific m�thylation. Both the reagent and the conditions of modification were found to be important in achieving a selective modification of fibrinogen.A possible interpretation of these results is the esterification of carboxylic acid groups in the fibrinogen with reduction in the prevailing carboxylate ion negative repulsion, thereby achieving an increased protein-protein interaction with a resulting polymerization.

Transient nitronic acid formation in the protonation of the carbanion of 2,2′,4,4′-tetranitrodiphenylmethane in acidic methanol

1987 ◽  
Vol 65 (8) ◽  
pp. 1980-1986 ◽  
Author(s):  
François Terrier ◽  
Jacques Lelièvre ◽  
Alain-Pierre Chatrousse ◽  
Robert Schaal ◽  
Patrick G. Farrell
Keyword(s):  
Carboxylic Acid ◽  
Solvent Effects ◽  
Rate Constants ◽  
Intrinsic Rate ◽  
Low Ph ◽  
The Other ◽  
Acid Formation ◽  
Other Hand ◽  
Protonated Species ◽  
Nitronic Acid

Rates of deprotonation of 2,2′,4,4′-tetranitrodiphenylmethane (1) by a variety of bases B− and of protonation of the resulting carbanion (2) by the conjugated acids BH in methanol have been measured at 20 °C. The Brønsted αBH coefficients for protonation of 2 by phenol and carboxylic acid buffers are equal to 0.58 and 0.43, respectively, as compared with αBH values of 0.59 and 0.52 respectively, for the same reactions in 50% H2O – 50% DMSO. On the other hand, a comparison of the intrinsic rate constants [Formula: see text] (in the Marcus sense) in the two solvents reveals that the effect of going from 50% H2O – 50% DMSO to methanol is to lower [Formula: see text] for the phenol reactions by a factor of 2 but to increase [Formula: see text]for carboxylic acid reactions by a factor of 5. It is shown that the solvent effects on αBH and [Formula: see text] can all be understood in the context of the Principle of Imperfect Synchronisation (PIS) recently proposed by Bernasconi. In addition, a fast equilibrium protonation of the carbanion 2 is found to precede its conversion to 1 at low pH, i.e., pH < 5. The resulting protonated species is suggested to be the nitronic acid 2,H arising from fast protonation of a para NO2 group of 2. The [Formula: see text] value associated with the ionization of 2,H is ∼4.25.

Über die Umsetzung von Benzylmalonylchlorid mit verschiedenen Natriumsalzen von Carbonsäuren / The Reaction of Benzylmalonyl Chloride with Sodium Salts of Carboxylic Acids

1972 ◽  
Vol 27 (5) ◽  
pp. 528-530 ◽  
Author(s):  
Helga Wittmann ◽  
Helmut Rathmayr
Keyword(s):  
Carboxylic Acid ◽  
Carboxylic Acids ◽  
Acid Chloride ◽  
Sodium Benzoate ◽  
Sodium Acetate ◽  
The Other ◽  
Carboxylic Acid Chloride ◽  
Sodium Salts ◽  
Other Hand ◽  
Sodium Phenylacetate

Benzylmalonyl Chloride reacts in the presence of sodium acetate in boiling benzene to give tribenzyl-phloroglucinol-triacetate, however with sodium chloroacetate to 3,5-dibenzyl-6-phenethylpyran-2,4-dion. In both cases trimerisation of benzylketene or benzylketene carboxylic acid chloride occurs. On the other hand, benzylmalonylchloride reacts with sodium benzoate and sodium phenylacetate via a dimeric benzylketene carboxylic acid chloride under the loss of phosgene to yield cyclopentadienyl derivatives.

scholarly journals Characterization of human heterophil hemolysins induced by Schistosoma mansoni infection

1983 ◽  
Vol 78 (3) ◽  
pp. 257-267 ◽  
Author(s):  
Munir Chamone ◽  
L. Alves Oliveira ◽  
P. M. Z. Coelho ◽  
G. Gazzinelli ◽  
A. Oliveira Lima ◽  
...  
Keyword(s):  
Heat Treatment ◽  
Schistosoma Mansoni ◽  
Hemolytic Activity ◽  
Gel Filtration ◽  
The Other ◽  
Other Hand ◽  
Classic Pathway

Heterophil antibodies could be detected in sera from normal or from patient with chronic schistosomiasis. Their hemolytic activities depend on the integrity of the complement classic pathway. The heterophil antibodies from patient sera presented a higher specificity for Schistosoma mansoni antigen preparations than those detected in normal sera. Most of the hemolytic activity observed in normal sera can be destroyed at 56ºC for 4 min. On the other hand, about 80% of the sera from infected patients are partially or totally resistant to this heat-treatment. The hemolytic activities of sera were eluted from a gel filtration column in different fractions of the first peak.

Abnormal Factor VIII Molecule in Mild Von Willebrand’s Disease

1979 ◽  
Author(s):  
R. Castillo ◽  
S. Maragall ◽  
F. Labal ◽  
J. Monteagudo ◽  
A. Ordinas
Keyword(s):  
Factor Viii ◽  
Concanavalin A ◽  
Gel Filtration ◽  
The Other ◽  
Agarose Gel ◽  
Immunoradiometric Assay ◽  
Von Willebrand's Disease ◽  
Von Willebrand’S Disease ◽  
Other Hand ◽  
Bleeding Episodes

In 22 cases with normal levels of the three entitles of factor VIII (VIII:C, VIIIR:AG and VIIIR:WF), FVIIIR:AG showed afaster anodal mobility and a lower concentration measured by immunoradiometric assay than that found with Laurell’s immunoelectrophoresis. Moreover, the factor VIII protein elution by agarose gel filtration of plasma and cryoprecipitate was delayed. Seven of those cases belong to a family in whom one of the member showed frequent bleeding episodes. In these individuals the FVIIIR;AG precipitation with Concanavalin A was determined and appeared decreased.On the other hand in 19 out of 20 subjects with classic moderate von willebrand’s disease (in whom the three entities of factor VIII were occasio nally reduced in parallele), FVIIIR:AG also showed a faster anodal uob{ 1-ty and the factor Vln protein elution by agarose gel filtration was delay ed.All these results suggest that patients with moderate von Willebrand’s disease show a qualitatively deficient factor VIII molecule synthesis, while the mildest cases may have normal levels of the three entities of the factor VIII.

Sign in / Sign up

Export Citation Format

Share Document