Characterisation of Serum Fibrinogen Degradation Products Using Gel Chromatography and Radioimmunoassay

1977 ◽  
Vol 37 (03) ◽  
pp. 471-476 ◽  
Author(s):  
S. M Ratky ◽  
A Sykes ◽  
E. D Cooke ◽  
Y. B Gordon
Keyword(s):  
Molecular Weight ◽  
Biological Activity ◽  
Venous Thrombosis ◽  
Molecular Species ◽  
Degradation Products ◽  
Gel Chromatography ◽  
Pulmonary Emboli ◽  
Serum Samples ◽  
Antigen Present

SummaryFibrin(ogen) degradation products (FDP) are detected by radioimmunoassay in the serum of all subjects. Some of these FDPs have considerable biological activity in vivo, and hence may play a significant part in the process of coagulation. Chromatography of serum samples from patients including those with pulmonary emboli and deep venous thrombosis indicates that the majority of non-clottable fibrin(ogen)-related antigen present is of high molecular weight- greater than that of fibrinogen – and that in pathological situations there is a quantitative increase not a qualitative alteration in the molecular species present.

scholarly journals Solubility and ADMET profiles of short oligomers of lactic acid

ADMET & DMPK ◽  
2020 ◽  
Author(s):  
Daniela Dascălu ◽  
Diana Larisa Roman ◽  
Madalina Filip ◽  
Alecu Aurel Ciorsac ◽  
Vasile Ostafe ◽  
...  
Keyword(s):  
Molecular Weight ◽  
Glucocorticoid Receptors ◽  
Degradation Products ◽  
Organic Anion ◽  
Toxicity Tests ◽  
Medical Applications ◽  
Eye Injuries ◽  
Toxicological Effects

<p class="ADMETkeywordsheading">Polylactic acid (PLA) is a polymer with an increased potential to be used in different medical applications, including tissue engineering and drug-carries. The use of PLA in medical applications implies the evaluation of the human organism's response to the polymer inserting and to its degradation products. Consequently, within this study, we have investigated the solubility and ADMET profiles of the short oligomers (having the molecular weight lower than 3000 Da) resulting in degradation products of PLA. There is a linear decrease of the molar solubility of investigated oligomers with molecular weight. The results that are obtained also reveal that short oligomers of PLA have promising pharmacological profiles and limited toxicological effects on humans. These oligomers are predicted as potential inhibitors of the organic anion transporting peptides OATP1B1 and OATP1B3, they present minor probability to affect the androgen and glucocorticoid receptors, have a weak potential of hepatotoxicity, and may produce eye injuries. These outcomes may be used to guide or to supplement in vitro and/or in vivo toxicity tests such as to enhance the biodegradation properties of the biopolymer.</p>

BINDING AND METABOLISM OF HEPARIN BY ENDOTHELIAL CELLS

1987 ◽  
Author(s):  
S Vannucchi ◽  
F Pasquali ◽  
P Bianchi-ni ◽  
M Ruggiero
Keyword(s):  
Molecular Weight ◽  
Endothelial Cells ◽  
De Novo ◽  
Degradation Products ◽  
Low Molecular Weight ◽  
Competition Binding ◽  
Capillary Endothelial Cells ◽  
A Site ◽  
Binding Sequence

In this study we show that bovineadrenal capillary endothelial cells(BACE) contain heparin (HP); this HP has been found associated with the cell surface (i.e; trypsin-removable^and intracellularly. How-ever, experiments with [ sjsodium sulfate labelling, demonstrate that BACE cells donot synthesize HP de novo, but they uptake it from serum. We have studied binding, uptake, and metabolism odifferent molecular weight-HPs: 13 Kd-HP from bovine source, 14 Kd-HP from porcine source, 4.5 Kd, and 2.5-HP fragments. Comparison among different HPs, was carried out by calculating the IC from competition curves for [3HJ- HP. Binding of labelled-HP to BACE cells was specificand saturable. Dextran sulfate and glycosaminoglycans did not compete for binding; only heparan sulfate showed some competition. Binding of different HPs was strictly dependent on their molecular weight; 2.5 Kd- HP was unable to bind to cells, although sulfation degree of this fragment and of unfractionated HP was almost identical. Therefore, we assume that a specific oligosaccharide sequence could be responsible for HP binding to BACE cells; this hypothetical "binding sequence" could then be lost in very low molecular weight-HP fragments. BACE cells are also able to internalize HP, and they release its low molecular weight degradation products into culture medium. Thus we suggest that endothelial cells might represent a site for the metabolism of endogenous and exogenous HP in vivo.

The Fibrin Assay Comparison Trial (FACT)

2001 ◽  
Vol 85 (04) ◽  
pp. 671-678 ◽  
Author(s):  
Sybille Zips ◽  
Hanimsah Ergül ◽  
Dieter Heene ◽  
Carl-Erik Dempfle ◽  
Keyword(s):  
Molecular Weight ◽  
High Molecular Weight ◽  
Degradation Products ◽  
Plasma Samples ◽  
Fibrin Degradation Products ◽  
Comparison Trial ◽  
Clinical Conditions ◽  
D Dimer

SummaryAlthough D-dimer has gained widespread clinical use as a parameter for detection of in vivo fibrin formation, the issue of standardization of D-dimer assays remains to be resolved. The FACT study was performed to generate basic data for development of calibrators and standard preparations.A set of 86 samples, including plasma samples from patients with DIC, DVT, and other clinical conditions, serial dilutions of pooled plasma samples, and plasma samples containing fibrinogen- and fibrin derivatives, were distributed to 12 manufacturers of D-dimer assays.D-dimer assays differ concerning specificity for crosslinked fibrin, and preference for either high molecular weight fibrin complexes, or low molecular weight fibrin degradation products. Terminal plasmin digests of fibrin clots for calibration produce aberrant results in some assays, especially those with preference for high molecular weight crosslinked fibrin derivatives. The best conformity is achieved by the use of pooled plasma samples from patients with high levels of D-dimer antigen in plasma. In vitro preparations containing a comparable composition of fibrin derivatives to clinical plasma samples may also serve as reference material.

scholarly journals Products of Metabolism and Processing of Lactic Acid Bacteria as Functional Ingredients

2018 ◽  
Vol 1 (1) ◽  
pp. 47 ◽  
Author(s):  
Antonina Ivanovna Kapustian ◽  
Natalia Cherno ◽  
Alexei Kovalenko ◽  
Kristina Naumenko ◽  
Igor Kushnir
Keyword(s):  
Amino Acids ◽  
Molecular Weight ◽  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
Organic Acids ◽  
Cell Walls ◽  
Degradation Products ◽  
Lactobacillus Delbrueckii ◽  
Low Molecular Weight ◽  
Gel Chromatography

Lactic acid bacteria (LAB) and bifidobacteria (BB) are unique substances that have a lot of biological and physiological effects. Structural components of LAB and BB – peptidoglycans, compounds of the muramylpeptide series, teichoic acids – have powerful immunological properties. Metabolites of LAB and BB – organic acids, hydrogen peroxide, bacteriocins, etc. – provide antagonistic activity, have an indirect impact on the immune system, reducing the antigenic load caused by pathogenic microorganisms. The expediency of peptidoglycans degradation of LAB and BB cell walls is substantiated. Low molecular weight products of the degradation can easily be absorbed and enter into biochemical processes, accelerating the expected functional-physiological effect. To obtain low-molecular products of peptidoglycans degradation, a combination of LAB and BB was used. The combination of LAB and BB is the sum of the test cultures of Lactobacillus acidophilus, Lactobacillus delbrueckii subsp. Bulgaricus, Bifidobacterium bifidum, Lactococcus cremoris, Streptococcus termophilus. Destruction of peptidoglycans of bacterial cell walls was carried out using a combination of disintegrating factors. The efficiency of destruction was determined by the accumulation of low molecular weight peptides (with molecular weight up to 1500 Da), amino acids and soluble protein in the disintegrate. It has been established that the highest accumulation of low molecular weight degradation products occurs when using autolysis followed by enzymatic hydrolysis during 180 min with the ratio of the enzyme : substrate 1 : 100. At the same time ≈ 53% of protein substances pass from insoluble to soluble state. The molecular weight of the obtained products is determined by the gel chromatography method. The qualitative and quantitative content of organic acids, amino acids and vitamins of group В in the hydrolysis products composition was investigated. It was shown that the obtained product possesses high biological effect in the experiment on animals.

Pulmonary Insufficiency in the Rat After Intravascular Coagulation and Inhibition of Fibrinolysis - Studies on Pulmonary Microcirculation and Pathophysiology

1979 ◽  
Author(s):  
B. Gerdin ◽  
T. Saldeen ◽  
H. Sandler ◽  
G. Sedin
Keyword(s):  
Molecular Weight ◽  
Mast Cells ◽  
Pulmonary Oedema ◽  
Degradation Products ◽  
Pulmonary Insufficiency ◽  
Low Molecular Weight ◽  
Oedema Formation ◽  
Intravascular Coagulation ◽  
Pulmonary Microcirculation

In posttraumatic pulmonary insufficiency microembolism of fibrin to the lungs and inhibited fibrinolysis have been given key-roles and the term "the delayed microembolism syndrome" has been designated to those cases. The pathogenetic mechanisms involved are only to a limited extent known. Those were investigated in a rat model. Fibrinolysis was inhibited with AMCA and intravascular coagulation induced by a 5-min injection of thrombin, 500 NIH/kg b.w.. Within 5 min a profound consumption of fibrinogen occurred and 125-la- belled fibrin was embolized to the lungs. At that time the alveolar circulation was decreased, vasoconstriction and platelet-red cell aggragates appeared as seen by in vivo microscopy and arterial P02 was diminished irrespective of fibrinolysis. After restitution for 1 h a progressive pulmonary insufficiency with P02 decrease and PC02 retention occurred. Trasylol® did not influence on pulmonary damage. Converting enzyme activity was not diminished. A retention of fibrin degradation products (fdp) occurred in the lungs and infusion of low molecular weight fdp aggravated pulmonary oedema. Lung mast cells were partly degranulated and mepyramine maleate slightly counteracted pulmonary oedema. It is concluded that release of vasoactive low molecular weight fdp probably plays a causal role in oedema formation; probably in part by releasing histamine from lung mast cells.

Unreliability of Current Serum Fibrin Degradation Product (FDP) Assays

1985 ◽  
Vol 53 (03) ◽  
pp. 301-302 ◽  
Author(s):  
P J Gaffney ◽  
M J Perry
Keyword(s):  
Monoclonal Antibodies ◽  
Serum Level ◽  
Degradation Product ◽  
Degradation Products ◽  
Polyclonal Antibodies ◽  
Fibrin Degradation Product ◽  
Serum Samples ◽  
Fibrinogen Degradation Products ◽  
A Value

SummaryPreviously, assays of fibrin-fibrinogen degradation products (FDP) had to be performed on serum samples. However, monoclonal antibodies (Mabs) are now available which permit the measurement of FDP directly in plasma. We have employed two Mabs, one monospecific for FDP originating from crosslinked fibrin and another panspecific for the FDP fraction, to determine normal FDP levels in plasma and serum. The monospecific Mab gave a value of 40 ng FDP/ml in plasma and 10 ng/ml in serum, while the serum level of FDP recorded using the panspecific Mab was >1000 ng/ml, at all the concentrations of thrombin employed. Similarly, when a solution of purified fibrinogen was treated with thrombin, the concentration of FDP present in the clot supernatant was >1000 ng/ml when assayed using the panspecific Mab. Thus during serum preparation as much as 75% of the native FDP is incorporated into the clot while in excess of 1000 ng/ml of laboratory generated FDP, probably incompletely polymerized fibrin, is measured using panspecific antisera. These data indicate that current FDP assays using polyclonal antibodies are not a reliable reflection of the FDP level generated in vivo. The use of FDP-specific Mabs which do not react with fibrinogen is recommended for future FDP assays performed directly on plasma.

scholarly journals Fibrin and fibrinogen degradation products with an intact D-domain C- terminal gamma chain inhibit an early step in accessory cell-dependent lymphocyte mitogenesis

Blood ◽  
1993 ◽  
Vol 81 (11) ◽  
pp. 3006-3014 ◽  
Author(s):  
SC Robson ◽  
R Saunders ◽  
LR Purves ◽  
C de Jager ◽  
A Corrigall ◽  
...  
Keyword(s):  
Molecular Weight ◽  
Degradation Products ◽  
Specific Binding ◽  
Lymphocyte Transformation ◽  
Mononuclear Leukocytes ◽  
Accessory Cell ◽  
Binding Studies ◽  
Gamma Chain

Abstract Although the low molecular weight degradation products of fibrinogen (FgDP) and fibrin (FbDP) are known to inhibit lymphocyte blastogenesis, the effect of purified macro-molecular FgDP and FbDP (molecular weight, 90 to 200 Kd) is unclear. We have examined the effect of these latter FgDP and FbDP and find that products that contain the D domain inhibit lymphocyte proliferation in response to T-cell mitogens, allogeneic mononuclear leukocytes, and anti-CD3 in vitro. Plasmic digestion of D1 in the absence of calcium with removal of the C-terminal end of the gamma chain or disruption of the gamma-gamma C-terminal cross-link site of D-dimer (DD) by puffadder venom (PAV-D) abrogates their inhibitory potential. Prior incubation of monocytes with DD or D1 inhibits subsequent lymphocyte transformation. Binding studies with radiolabeled DD and PAV-D confirm that monocytes interact only with DD. This specific binding may be competitively inhibited by monoclonal antibodies to CD11b/CD18 or by peptide analogues of the C-terminal gamma chain of fibrinogen that mimic the adhesion recognition site of integrins. We postulate that DD and D1 bind to CD11b/CD18 on adherent monocytes and modulate lymphocyte activation. These products are typically present in the plasma of patients with disseminated intravascular coagulation with sepsis and could therefore influence inflammatory processes in vivo.

Iron availability from meat

1978 ◽  
Vol 39 (3) ◽  
pp. 631-638 ◽  
Author(s):  
T. Hazell ◽  
D. A. Ledward ◽  
R. J. Neale
Keyword(s):  
Molecular Weight ◽  
Degradation Products ◽  
Gel Filtration ◽  
Rat Muscle ◽  
Muscle Extract ◽  
Freeze Dried ◽  
Absorption Studies ◽  
Fe Complexes

1. The distribution of radioactive iron in59Fe-labelled rat muscle extract was determined using ge filtration. This showed that most (approximately 70%) of the radioactivity was associated with the heamatin compounds; myoglobin and haemoglobin.2. Raw beef and freeze-dried rat muscle were digested in vitro, under simulated physiological conditions, and after centrifugation the supernatants fractionated by gel filtration. The soluble products were haematin Fe complexes of molecular weight above 10 000 and non-haematin Fe compounds of molecular weight below 6000, the major products being the non-haematin Fe complexes. The soluble compounds were also separated by dialysis and, in rat muscle, it was found that the low-molecular-weight non-haematin compounds accounted for more than 80% of the total soluble iron.3. In vivo absorption studies with rats showed the Fe in a digested muscle dialysate to be more readily absorbed than that from an aqueous muscle extract which itself was more readily absorbed than the Fe from whole blood.4. It may not, therefore, be the haemoproteins per se which are responsible for the high availability of Fe in meat, but rather the nature of their degradation products, formed by digestion within the meat environment.

Pulmonary Insufficiency in the Rat After Intravascular Coagulation and Inhibition of Fibrinolysis - Studies on Pulmonary Microcirculation and Pathophysiology

1979 ◽  
Author(s):  
B. Gerdin ◽  
T. Saldeen ◽  
H. Sandler ◽  
G. Sedin
Keyword(s):  
Molecular Weight ◽  
Mast Cells ◽  
Pulmonary Oedema ◽  
Degradation Products ◽  
Pulmonary Insufficiency ◽  
Low Molecular Weight ◽  
Oedema Formation ◽  
Intravascular Coagulation ◽  
Pulmonary Microcirculation

In posttraumatic pulmonary insufficiency microembolism of fibrin to the lungs and inhibited fibrinolysis have been given key-roles and the term “the delayed microembolism syndrome” has been designated to those cases. The pathogenetic mechanisms involved are only to a limited extent known. Those were investigated in a rat model. Fibrinolysis was inhibited with AMCA and intravascular coagulation induced by a 5-min injection of thrombin, 500 NIH/kg b.w.. Within 5 min a profound consumption of fibrinogen occurred and 125-la-belled fibrin was embolized to the lungs. At that time the alveolar circulation was decreased, vasoconstriction and platelet-red cell aggragates appeared as seen by in vivo microscopy and arterial PO2 was diminished irrespective of fibrinolysis. After restitutio for tha progressive pulmonary insufficiency with PO2 decrease and PCO2 retention occurred. Trasylol did not influence on pulmonary damage. Converting enzyme activity was not diminished. A retention of fibrin degradation products (fdp) occurred in the lungs and infusion of low molecular weight fdp aggravated pulmonary oedema, l.ung mast cells were partly degranulated and mepyramine maléate slightly counteracted pulmonary oedema. It is concluded that release of vasoactive low molecular weight fdp probably plays a causal role in oedema formation; probably in part by releasing histamine from lung mast cells.

Size heterogeneity of immunoreactive prolactin in patients with prolactinoma

1987 ◽  
Vol 114 (4) ◽  
pp. 475-482 ◽  
Author(s):  
B. Allolio ◽  
A. Hoeppener ◽  
U. Leonhardt ◽  
U. Deuβ ◽  
W. Winkelmann
Keyword(s):  
Tumour Size ◽  
Serum Prolactin ◽  
Gel Chromatography ◽  
Freezing And Thawing ◽  
Serum Samples ◽  
Elution Pattern ◽  
Long Term Storage ◽  
Repeated Freezing

Abstract. We investigated the chromatographic pattern of serum prolactin in 41 patients with prolactinoma and correlated the distribution of immunoreactive prolactin with the clinical variables sex, tumour size, age, and response to bromocriptine therapy. In addition, the effect of long-term storage and repeated freezing and thawing on the different molecular weight forms of prolactin was evaluated. Gel chromatography (column 100 cm × 1.5 cm) was performed in 0.1 mol/l phosphate buffer, pH 7.5, using Ultrogel ACA 54 (LKB). No correlation of age or the response to drug therapy to the elution pattern of prolactin was found. Females showed a higher percentage of big prolactin than males (10.4 ± 1.2% vs 6.8 ± 0.7%, x̄ ± sem, P <0.05) and patients with microprolactinomas too had a higher percentage of big prolactin than those with macroprolactinomas (11.3 ± 1.8% vs 7.7 ± 0.7%, P <0.05). Serum samples kept frozen for more than 2 years showed a higher percentage of bigbig prolactin (P < 0.01) than samples stored for less than 12 months suggesting formation in vitro. However, examination of fresh samples prior to freezing also demonstrated bigbig prolactin, indicating that bigbig prolactin circulates in vivo. Repeated freezing and thawing of bigbig prolactin led to almost complete interconversion to little prolactin without any increase in immunoreactivity. This finding supports the concept that bigbig prolactin represents little prolactin loosely associated to a carrier molecule.

Sign in / Sign up

Export Citation Format

Share Document