Changes in the Electrophoretic Patterns of Euglobulin Fractions Following Activation of the Fibrinolytic System by Exercise

1971 ◽  
Vol 49 (7) ◽  
pp. 672-677 ◽  
Author(s):  
Valerie McAlpine ◽  
Susan Milojevic ◽  
Frank C. Monkhouse

A study was made of changes in blood fibrinolytic activity in a group of young male university students following varying levels of exercise. The subjects were divided into two groups. Those partaking in regular athletic programs were classified as athletes and those not as nonathletes. There was an increased fibrinolytic activity with increased severity of exercise in all subjects. Several of the nonathletic group showed a much greater maximum effect than their athletic counterparts, and all nonathletes showed the effect at much lower levels of exercise. Increase in fibrinolytic activity correlated well with increase in pulse rate. When euglobulin fractions from the plasma samples were subjected to microzone electrophoresis, there was a band change in the region of the beta globulins in many of the samples following exercise. This band change is apparent only when fibrinogen is present in the euglobulins and appears to be due to fibrinogenolysis as a result of the activation of the fibrinolytic system. These band changes were observed when the specific activity was as little as 0.058 unit of plasmin per milligram protein. Our results show that electrophoretic pattern changes of euglobulin fractions is a relatively sensitive and rapid method for detecting activation of the fibrinolytic system.

1992 ◽  
Vol 67 (06) ◽  
pp. 697-701 ◽  
Author(s):  
J J Emeis ◽  
A Brouwer ◽  
R J Barelds ◽  
M A Horan ◽  
S K Durham ◽  
...  

SummaryAged rats are more susceptible to endotoxin-induced effects, including microthrombosis and platelet aggregation, than are young rats. To investigate whether changes in the fibrinolytic system might be involved, we investigated the fibrinolytic activity in plasma euglobulin fractions and tissues (lung and heart) of young (6-months old) and aged (24-months old) rats under baseline conditions and after challenge with endotoxin. Aged rats had lower plasma levels of tissue-type plasminogen activator (t-PA) and of urokinase-type PA (u-PA) activity. PA inhibitor (PAI) activity was higher in the plasma of aged rats, as was t-PA activity in lung and heart.Rats were treated with either a low dose (1 μg/kg) or a high dose (10 mg/kg) of endotoxin. Both treatments induced a transient phase of increased blood fibrinolytic activity, as evidenced by higher levels of tissue-type plasminogen activator (t-PA) activity and decreased levels of PA inhibitor (PAI) activity. Over time, the fibrinolytic activity decreased, probably due to increased levels of PA inhibitor.Both the early increase in t-PA activity, and the subsequent increase in PAI activity, were more pronounced in the aged rats, as compared with the younger rats, after the high dose of endotoxin. The aged rats also responded to an injection of interleukin-1β or tumor necrosis factor-α with a larger increase of PAI activity than did the younger rats.Together the data suggest that, compared to young rats, aged rats have a decreased base-line plasma fibrinolytic activity, while their fibrinolytic system is more responsive to challenge by endotoxin and cytokines.


1963 ◽  
Vol 10 (01) ◽  
pp. 120-132 ◽  
Author(s):  
E. S Olesen

SummaryTreatment of serum with bentonite led to a reduced content of inhibitors of trypsin and urokinase in the isoelectrically precipitated euglobulin, and removed fibrinolytic agents and precursors from serum. Bentonite-treated serum added to untreated serum reduced precipitation of the above inhibitors, and presumably also precipitation of inhibitors against a plasminogen activator of serum.Bentonite-treated serum (whether from pig, ox, guinea-pig, or man), added to untreated guinea-pig serum, produced fibrinolytic activity on isoelectric precipitation of the mixture; the activity of the euglobulin was due to an activator of plasminogen as well as an active protease, probably plasmin. The described effects of bentonite-treated serum are similar to those previously reported for anionic polyelectrolytes. Possible mechanisms are discussed.The “non-specific” activation of fibrinolytic activity by means of bentonite emphasizes that guinea-pig serum [which is characterized by a high potential for “nonspecific” activation of its fibrinolytic system Olesen (1962)] contains all the elements required for the formation of an activator of plasminogen, and thus the activation of its plasminogen to plasmin.


2016 ◽  
Vol 41 (3) ◽  
Author(s):  
Birkan Slem ◽  
Yüksel Gezgin ◽  
Rengin Eltem

AbstractObjective: To screen fibrinolytic enzyme-producing Bacillus isolates (n=210) and to characterize of thermostable fibrinolytic enzyme from Bacillus amyloliquefaciens EGE-B-2d.1 that had the highest level of fibrinolytic activity together with the highest thermostability.Methods: Firstly, a total of 210 isolates were screened for their fibrinolytic enzyme production. The potent fibrinolytic enzyme producing isolates were evaluated for the thermostability of their fibrinolytic enzymes and one isolate showing prominent fibrinolytic activity was identified as molecular. Fermentation process was carried out on the isolate that had both the highest level of fibrinolytic activity and enzyme thermostability. The thermostable fibrinolytic enzyme from this isolate was then purified and characterized.Results: The fibrinolytic enzyme activities of 21 Bacillus sp. isolates in Nutrient Yeast Salt Medium were found to be in the range of 0.176-1.734 U/ml. The fibrinolytic activity of the enzyme purified from the culture supernatant of Bacillus amyloliquefaciens EGE-B-2d.1 was relatively stable at pH 7.0-11.0 for 24 h and also showed stability at a temperature of 60°C for 60 min. The enzyme degraded the fibrin clots by direct fibrinolysis. The specific activity and the molecular weight of the purified enzyme were estimated to be 44.46 units/mg protein and 30 kD respectively.Conclusion: The thermostable fibrinolytic enzyme from Bacillus amyloliquefaciens EGE-B-2d.1 was purified and characterized. This enzyme might also be used as a natural agent for oral fibrinolytic therapy or thrombosis prevention.


Weed Science ◽  
2010 ◽  
Vol 58 (3) ◽  
pp. 258-264 ◽  
Author(s):  
Syuntaro Hiradate ◽  
Kenji Ohse ◽  
Akihiro Furubayashi ◽  
Yoshiharu Fujii

The allelopathic potential of a plant has been evaluated on the basis of two indicators: specific activity, which is the specific concentration of the allelochemical to exert a half-maximum effect on a receiver plant (EC50), and total activity in a plant, which is the ratio of the concentration of an allelochemical in the producing plant to its EC50. In the present study, a new indicator, total activity in a soil, which takes into account the effects of a soil on the allelopathy activity, is proposed because allelopathic activity is affected by the presence of soils. The total activity in a soil was calculated by multiplying the “total activity in a plant” with a “soil factor.” In this calculation, we assumed simplified cases for comparison, such that the allelopathic plant materials are evenly incorporated in the soils and the allelochemicals are released from the plant materials to the soils at a constant rate. We conducted bioassay experiments in the presence and absence of soils and cited some published data to calculate the specific activities and total activities in a plant and in a soil. The results indicated that the allelopathies of buckwheat caused by (+)-catechin, Leucaena leucocephala by L-mimosine, Xanthium occidentale by trans-cinnamic acid, and Brassica parachinensis by cis-cinnamic acid were not significant in a volcanic ash soil, an alluvial soil, and a calcareous soil, but the allelopathy of sweet vernalgrass caused by coumarin and Spiraea thunbergii by cis-cinnamoyl glucosides was highly effective in those soils. The allelopathies of Juglans species caused by juglone plus juglone precursors and Mucuna pruriens by L-DOPA would depend highly on the soil types. Although some limitations exist for this approach, the total activity approach would allow for a better quantitative estimation of the allelopathic potential of plant materials in soils.


1966 ◽  
Vol 44 (1) ◽  
pp. 95-101 ◽  
Author(s):  
J. R. Beaton ◽  
A. J. Szlavko ◽  
J. A. F. Stevenson

The effect of various factors on excretion of a lipid-mobilizing activity in FMS IA (anorexigenic) and in FMS IB (fat-mobilizing) by the fasting rat has been investigated. During fasting, the greatest excretion of such activity in FMS IA and FMS IB occurred in the first 24 hours and diminished thereafter up to 72 hours; and the specific activity of FMS IB was greatest in the first 24 hours whereas that of FMS IA was constant throughout. The hypothalamicobese rat excretes FMS IA and FMS IB in greater than normal amounts. The alloxan-diabetic rat excretes less total activity of FMS IA and IB than do control animals. Young male rats excrete greater amounts of FMS IB, but not of FMS IA, than do adult rats, the greatest excretion per 100 g body weight being observed at approximately 37 days of age. At 27 days of age (prepuberty), male rats excreted a greater total activity of FMS IB but not of FMS IA than did female rats. At 90 days of age (post-puberty), there was no apparent sex difference in the amount of total activity of FMS IB excreted per rat, but when expressed per 100 g body weight, females excreted more FMS IB than did males.


1984 ◽  
Vol 221 (2) ◽  
pp. 465-470 ◽  
Author(s):  
R J Edwards ◽  
D C Watts

The effect of partially purified ‘creatine kinase conversion factor’ on rabbit muscle creatine kinase is shown to be that of a carboxypeptidase, removing the C-terminal lysine residue from both subunits. These changes fully explain the three-banded electrophoretic patterns of the partially and the fully modified rabbit and human enzymes. The factor also produces a similar electrophoretic pattern with haemoglobin A; comparison with the effects of carboxypeptidases A and B permits the inference that the C-terminal residues of both alpha- and beta-subunits are removed. Small synthetic peptides are poor or non-substrates. A low activity with hippuryl-L-lysine may be due to contamination of the preparation with carboxypeptidase N. The possibility has been excluded that the action of conversion factor on creatine kinase involves modification of the protein thiol groups. Mr, substrate-specificity, pH-activity profile and the effects of metal ions distinguish creatine kinase conversion factor from carboxypeptidases A, B and N. On the basis of this evidence it is proposed to give the conversion factor the provisional name of carboxypeptidase K.


1981 ◽  
Author(s):  
M Yamauchi ◽  
H Takei ◽  
T Seya ◽  
Y Oguma ◽  
T Murakoshi ◽  
...  

ABy means of SDS-PAGE (3.3% gel), Fbg heterogeneity originated from partial degradation of Aα chain was studied. Comparison of electrophoretic patterns of plasma and corresponding serum made it possible to identify 2 major Fbg bands designated as high-molecular-weight Fbg (HMW, MW 350,000) and low-molecular-weight Fbg (LMW, MW 310,000). LMW comprised 28×2% (mean×S.D) of total Fbg (HMW+LMW) in healty subjects. The elevation of fibrinolytic activity did not accompany the increase of percentages of LMW in various diseases, even in cirrhotic patients whose levels of α2;PI were low. In DIC patients percentages of LMW were decreased extremely (12×6%, mean×SD). Samples from animal experimental models of DIC exhibited the same pattern of Fbg heterogeneity as that of DIC patients.UK was added to the purified Fbg in vitro. On the earliest stage of the fibrinogenolysis. 2 bands appeared newly on SDS-PAGE, while the bands of HMW and LMW were decreased. One of these new bands (Band 1) corresponded with a major compornent of Fraction 1-9 of Mosesson. It was located in the slightly anodal position (MW 300,000) from LMW band. Another band (MW 270,000) migrated between Band 1 and the band of Frag X. The same pattern of Fbg heterogeneity was observed in patients recieving large dose of UK. After cessation of UK treatment these new bands disappeared, while the bands of HMW was increased extremelThese findings suggest that HMW is a freshly synthesized Fbg and that unknown mechanism without plasmin may present for the conversion HMW to LMW.


1959 ◽  
Vol 196 (2) ◽  
pp. 431-435 ◽  
Author(s):  
S. N. Kolmen ◽  
D. R. Celander ◽  
M. Mason Guest

Activation of the immunologic system either in vitro or in vivo results in nearly complete consumption of complement without activation of the fibrinolytic system. However complement activity decreased upon the induction of fibrinolytic activity even in the absence of an antigen-antibody reaction. Presumably this is due to proteolysis of some part of the complement complex since complement activity was found to be destroyed in the presence of small quantities of purified fibrinolysin. These observations are consistent with the conclusion that a) the components of complement and those of the fibrinolytic system are separate and discrete entities; b) that reactions involving complement fixation and activation do not directly influence the fibrinolytic system; and c) that activation of the fibrinolytic system results in decreases in complement through proteolysis of one or more of its components by the fibrinolysin which develops.


2019 ◽  
Vol 22 (2) ◽  
pp. 114-121
Author(s):  
Marco Giraldi ◽  
Saverio Paltrinieri ◽  
Paola Scarpa

Objectives The purpose of this study was to describe the electrophoretic patterns of proteinuria in cats at risk of and cats with chronic kidney disease (CKD), and to investigate whether the presence of high-molecular-weight (HMW) and low-molecular-weight (LMW) proteins were associated with CKD, proteinuria and/or disease progression. Methods Healthy cats at risk of developing renal disease (n = 17) and cats affected with CKD at different stages (n = 22) were prospectively enrolled and sampled over time. Seventy urine samples were included and assayed with a commercially available sodium dodecyl sulfate–agarose gel electrophoresis (SDS-AGE) method. Each sample (gel lane) was inspected to identify albumin, HMW and LMW proteins, and an electrophoretic pattern (albuminuria, glomerular, tubular, mixed or negative) was assigned accordingly. Fisher’s exact test was used to assess the distribution of HMW and LMW proteins in cats grouped according to International Renal Interest Society stage and to the magnitude of proteinuria, and to assess if HMW and LMW proteins at the time of inclusion were associated with the development and progression of CKD. Results In samples of cats at risk, the most common pattern was glomerular (84.6%); glomerular pattern was also common in cats with CKD (54.2%), although mixed proteinuria and tubular proteinuria were also present (29.5% and 11.4%, respectively). The presence of LMW proteins was associated with CKD ( P <0.0001) and to a urine protein:creatinine ratio >0.2 ( P = 0.025). Both HMW and LMW proteins were not associated with progression of CKD within 6 months (n = 14). Conclusions and relevance Our results showed that HMW proteinuria is common in healthy cats at risk of developing CKD, although the pathological significance needs to be confirmed. The detection of LMW proteins in urine of cats suspected to be affected by CKD, especially in non-azotaemic, non-proteinuric or borderline proteinuric cats, suggests the presence of kidney damage.


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