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While the prevalence of cancer-associated thrombosis (CAT) is high in cancer patients, its molecular mechanisms have not been fully elucidated. Moreover, the risks of recurrent CAT events and mortality remain high in cancer patients despite the introduction of anticoagulant/antiplatelet therapy. Here, we discuss the possibility that increased plasmin activity driven by anticoagulant/antiplatelet treatment might be the major mechanism responsible for the activation of an excess of cancer-derived transforming growth factor-beta (TGF-β) originating from cancer cells and the tumour microenvironment. Hence, high coagulation and fibrinolysis rates in cancer patients may be linked to high rates of TGF-β activation, especially the excess of TGF-β derived from cancer cells. In turn, high TGF-β activation could contribute directly to maintaining high thrombotic risk and CAT recurrence in cancer patients since TGF-β signalling increases gene expression and secretion of the fibrinolysis inhibitor plasminogen activator inhibitor 1 (PAI1). Thus, TGF-β could directly contribute to the high number of deaths among patients with cancer experiencing CAT, despite anticoagulant/antiplatelet treatment. In a longer-term perspective, increased TGF-β activation, by supporting a pro-coagulant cancer microenvironment, might also accelerate cancer progression. This review aims to discuss the published evidence that might support the scenario described above, and to put forward the hypothesis that cancer patients experiencing CAT events would largely benefit from anti-TGF-β therapy. [ABSTRACT FROM AUTHOR]

Introducing Holstein cows on Qinghai-Tibetan Plateau is a potential solution to enhance local milk production. However, the relationship between milk quality and altitude in China remains unknown. Therefore, the components and plasmin (PL) system of raw milk from different altitudes (sea level, 1600, 2700, and 3800 m) were investigated. The daily milk production of Holstein cows and PL activity decreased as the altitude increased. However, the components content of raw milk, plasminogen (PLG)/PL ratio, activities of PLG and plasmin activator (PA) increased with altitude. The pasteurization resulted a significant decrease in PA activity of all milk and a significant increase in PL activity in milk collected at higher altitudes (2700 and 3800 m), suggesting the pasteurization was unsuitable for preserving milk at higher altitudes. This study offered references for the production and storage of milk after introducing Holstein cows on Qinghai-Tibetan Plateau.

Pulsed electric field (PEF) processing was investigated as an alternative dairy preservation technology that would not compromise quality yet maintain safety. PEF processing of raw whole milk (4% fat) was conducted at two processing conditions (30 kV/cm, 22 μs, at either 53 or 63 °C outlet temperature) and compared with two thermal treatments (15 s, at either 63 or 72 °C) and a raw milk control and replicated twice. Milk bottles (2 L) from each treatment were incubated for two weeks, at 4 and 8 °C, and assessed for total plate count, pH, colour, rennetability, plasmin activity and lipid oxidation. The microbial quality of the thermal (72 °C/15 s) and PEF (63 °C) were similar. A drop in pH occurred after a change in counts was observed. Rennetability was not different between the treatments. Short chain acids dominated the volatile profile of the milk samples. Concentration of volatiles derived from microbial activity, namely 2,3-butanedione, acetic acid and other milk lipid derived short chain free fatty acids (e.g. butanoic and hexanoic acids), followed the trend of microbial activity in milk samples. Industrial relevance Research on the application of PEF to control spoilage and pathogenic microorganisms and enzyme systems in milk spans a wide array of processing equipment and reaction conditions. While industrial scale PEF processing of liquid milk for preservation and improved quality seems generally possible, substantiation of lower thermal damage under safe and scalable PEF conditions is required to enable economic feasibility. [ABSTRACT FROM AUTHOR]

The aim of this study was to evaluate the influence of shortening the dry period of Swedish dairy cows on plasmin activity and casein composition in milk. Swedish Holstein and Swedish Red cows, 45 in total, were assigned to a dry period of either 4 or 8 wk. Milk samples were taken 10 and 5 wk prepartum, and 6 and 12 wk postpartum. Plasmin activity and plasminogen activity were measured with a spectrophotometric assay. Casein composition was measured by capillary zone electrophoresis. Prepartum plasminogen activity increased by 22% between 10 and 5 wk prepartum, whereas no change in plasmin activity was observed during the same period. Cows with a 4-wk dry period had 61% higher plasmin activity in postpartum milk than cows with an 8-wk dry period. Cows of third or greater parity tended to have a stronger increase in plasmin activity as a result of applying a short dry period than cows of second parity. Although the αS1- and β-casein fractions declined with increasing plasmin activity, no dry period effects were found. Based on postpartum differences in plasmin activity, it was concluded that particularly multiparous cows require more than 4 wk between lactations for recovery of the mammary epithelium. Changes in casein composition as an effect of plasmin activity are not expected to have a great effect on processing quality of milk, although future work is needed to verify this. [ABSTRACT FROM AUTHOR]

Gastrodia rhizome, a dried and steamed tuber of Gastrodia elata Blume (Orchidaceae), has been traditionally used in Korea, China and Japan for the treatment of neurological and nervous disorders such as headaches, dizziness, vertigo and convulsive illnesses. The ethyl acetate and water extracts of G. elata stimulated plasmin activity. The active ethyl acetate fraction was subjected to centrifugal partition chromatography (CPC) with a two-phase solvent system, composed of n-hexane-ethyl acetate-methanol-water (3:7:4:6, v/v) followed by semi-preparative HPLC purification to separate active compounds and the water fraction was purified by Diaion HP-20 resin and semi-preparative HPLC. In ethyl acetate extract, 4-hydroxybenzyl alcohol (1), 4-hydroxybenzoic acid (2), 4-hydroxybenzaldehyde (3), 4-ethoxymethylphenol (4), 4,4′-oxy bis(methylene)diphenol (5) and 4,4′-methylenediphenol (6) were obtained with high purities. Parishin (7) and parishin B (8) were isolated from water extract. Among isolated compounds, 4-hydroxybenzyl alcohol (1), 4-hydroxybenzaldehyde (3) and 4-ethoxymethylphenol (4) significantly stimulated plasmin activity. Copyright © 2015 John Wiley & Sons, Ltd. [ABSTRACT FROM AUTHOR]

An isoindolone derivative, Fungi fibrinolytic compound ( R)-2,5-bis((2 R,3 R)-2-(( E)-4,8-dimethylnona-3,7-dien-1-yl)-3,5-dihydroxy-2-methyl-7-oxo-3,4,7,9-tetrahydropyrano[2,3- e]isoindol-8(2 H)-yl)pentanoic acid (FGFC1, Fungi fibrinolytic compound 1), was isolated from a rare marine microorganism strain Stachybotrys longispora FG216. The structure of FGFC1 was elucidated by 1H NMR, 13C NMR, IR, and MS data; moreover, it was also evaluated for fibrinolytic activity in vitro and in vivo. The results showed that 0.1-0.4 mmol/L of FGFC1 could stimulate generation of plasmin activity (increased by 2.05-11.44 folds) by measuring Glu-plasminogen and Lys-plasminogen activation in vitro. The experiment of fluorescein isothiocyanate (FITC)-fibrinogen degradation indicated that the effect of FGFC1 on fibrinolytic activity was mediated by plasminogen and scuPA. In addition, FGFC1 (10 mg/kg) could dissolve most of pulmonary thrombus of Wistar rat in vivo. It is possible that FGFC1 is a potential thrombolytic agent in the future. [ABSTRACT FROM AUTHOR]

Purpose: Aronia melanocarpa has an extremely high content of procyanidins and anthocyanins. The multidirectional benefits of consumption of these berries are widely reported. Although numerous studies confirmed the influence of polyphenols on various stages of hemostasis, the exact mechanism of this phenomenon is not understood. The aim of our study was to evaluate the in vitro effect of A. melanocarpa extract on various parameters of hemostasis. Methods: Adenosine 5′-diphosphate (ADP)-induced aggregation was measured with turbidimetric method. Spontaneous and ADP-activated platelet adhesion were investigated using a colorimetric method. The global assay of coagulation and fibrinolysis was performed with the use of optical clotting and lysis (CL) test. Thrombin (0.5 IU/mL) and tissue plasminogen activator (60 ng/mL) were used to obtain a CL curve. The activity of thrombin and plasmin was determined by means of chromogenic substrate (S-2238, S-2251) Results: The aronia extract contributed to the reduction in spontaneous and ADP-activated platelet adhesion. A significant increase in overall potential of CL as well as significant changes in key parameters of these processes ( T-thrombin time, F-initial plasma clotting velocity, and L-maximum lysis) was reported. Chokeberry extract significantly inhibited the amidolytic activity of thrombin and plasmin. Conclusion: Our in vitro findings indicate a complex mechanism of influence of chokeberry polyphenols on platelet activity and the overall potential of CL. We confirmed that chokeberry inhibits the amidolytic activity of thrombin. It was demonstrated for the first time that chokeberry polyphenols inhibit the amidolytic activity of another serine protease, i.e., plasmin, which is the main fibrinolytic enzyme. Furthermore, our research points out a significant contribution of other plasma components and fibrinogen in the modulation of hemostasis by polyphenols. [ABSTRACT FROM AUTHOR]

Abstract: In the present study, the relationship between milk somatic cell counts (SCC), nitrogen fractions, individual casein percentage distribution, plasmin-plasminogen activities and proteolysis, and the influence of lactation stage and lactation number in dromedary and cow milks were compared. Dromedary milk appeared to contain a lower amount of total and pH 4.6-soluble nitrogen than cow milk. The major protein components in milks were caseins, which accounted for ∼70% of total proteins for both species, β- and αs1-being the predominant caseins in both milk kinds. In dromedary milk, β-casein increased with lactation number but decreased as SCC increased. Total protein, αs2-casein and SCC decreased during lactation in dromedary milk. The dromedary is characterized by a relatively low content of κ-casein. Plasmin plus plasminogen activity is higher in cow milk compared to dromedary milk. The plasmin activity increased as SCC increased in cow milk but not in dromedary milk. [Copyright &y& Elsevier]

• Enzymatic hydrolysis of a specific substrate for plasmin activity is studied. • The effect of peptide sequence on enzyme activity was observed. • APPPPAKAPPPPC was the superior peptide sequence with optimal alignment and conformation. • A new SERS-based sensitive method was developed for the detection of plasmin activity. • The method was successfully used for the detection of plasmin activity in real milk samples. In this study, a new surface-enhanced Raman scattering (SERS)-based method has been developed for the detection of plasmin activity. Firstly, different peptide sequences, which are specific to plasmin, were examined. Then, SERS substrates were prepared by chosen peptide substrate. Enzyme activity was determined by pursuing the reduction of DTNB band at 1331 cm−1 with Raman spectroscopy. The reduction in SERS intensity was related to the plasmin activity, and changes in SERS intensity vs. plasmin concentration graph was obtained. Limit of detection (LOD) and limit of quantification (LOQ) values were calculated as 2.14 U/mL and 6.42 U/mL, respectively. Intra- and inter-day repeatability results were determined as 1.45% and 1.47% relative standard deviation (RSD). Also, recovery of the method was determined for the plasmin spiked milk samples. The results demonstrated that the proposed method could be successfully used to detect the plasmin activity in milk samples. [ABSTRACT FROM AUTHOR]

Background/Aims: The pathogenesis of alcoholic liver disease (ALD) remains uncertain. Fibrin production and degradation are altered in experimental liver injury. We have recently identified increased expression of a number of genes (annexin A2 (ANXA2), p11, tPA and PAI-1) that implicate fibrinolysis in ALD progression. Aim of our study was to study the direct effect of alcohol on fibrinolysis and plasmin activity in hepatic cell lines and in vivo. Methods: Expression of pro- and anti-fibrinolytic genes was determined in liver biopsies from patients with progressive ALD and in HepG2, Huh7 and LX-2 cells exposed to alcohol. The functional effects on fibrinolysis and plasmin activities were determined. C57BL6 female mice were given a single dose of alcohol and serum and liver triglyceride content and serum plasmin activity determined. Results: Alcohol induced a significant up-regulation of ANXA2, PLG, PAI-1 and p11 in human ALD, cell lines and in mice exposed to alcohol. Up-regulation of ANXA2 and p11 was inhibited by the alcohol dehydrogenase inhibitor 4-methylpyrazole. Fibrinolysis and plasmin were increased in HepG2 and LX-2 cells by 10mM alcohol and was inhibited by ANXA2 blocking antibody. Plasmin also increased in mice given a moderate dose of alcohol. By contrast, there was striking up-regulation of PAI-1 in mice given a high dose of alcohol with associated decrease in plasmin. Conclusions: Alcohol directly alters hepatic expression of pro- and anti-fibrinolytic genes in a dose dependent manner with low dose promoting fibrinolysis and high dose inhibiting fibrinolysis. After a large dose of alcohol in vivo, the dominant effect was up-regulation of hepatic PAI-1 with suppression of plasmin. The effect of alcohol on fibrinolysis and plasmin is mediated in part by ANXA2. Alcohol directly influences hepatic pathways of fibrinolysis that may contribute to ALD. [Copyright &y& Elsevier]

Abstract: Rennet pastes produced by lambs subjected to three different feeding systems (mother suckling [MS], artificial rearing [AR], and artificial rearing with Lactobacillus acidophilus supplementation [ARLB]) and slaughtered at two different ages (20 and 40d) were used for the manufacture of Pecorino foggiano cheese. Composition and proteolysis during ripening of Pecorino foggiano cheese (four replicates batches) were analyzed. Proteolysis was greater in cheeses made with rennet pastes from lambs slaughtered at 20d, as shown by analysis of nitrogen fractions (water-soluble N and proteose peptones). Supplementation of milk substitute with L. acidophilus may have influenced the growth dynamics of lactic acid bacteria in the rennet pastes, with positive effects on levels of lactobacilli in cheese at the beginning of the ripening time. Lower pH values in ARLB cheese during ripening, together with higher cell loads, suggest that supplementation of milk replacer with L. acidophilus resulted in higher proteolytic activity, as also confirmed by the composition of the pH 4.6—insoluble nitrogen fraction. No differences were found in total concentration of free amino acids among the experimental cheeses; phenylalanine, isoleucine, leucine, lysine were found at the highest levels. The addition of probiotic bacteria to milk substitute in lamb rearing appears to give good-quality lamb rennet paste. [Copyright &y& Elsevier]

Abstract: Havarti 45+ cheese was manufactured from milk concentrated 1.8–4.6-fold by ultrafiltration (UF) and from normal milk, and the effect of concentration factor on plasmin activity and subsequent proteolysis in cheese during ripening was examined. There was decreased plasmin activity and a reduced rate of proteolysis of α S2-casein and β-casein in the UF-cheeses, compared with the corresponding controls, independent of concentration factor. The decreased plasmin activity and slower breakdown of α S2-casein and β-casein in UF-cheeses compared with traditional cheeses can be partly explained by the inclusion of an increased amount of plasmin inhibitors into the UF-cheeses. However, it is suggested that the differences in plasmin activity and proteolysis arise mainly as a result of inactivation of the plasminogen activation system during UF-concentration, due to a combination of time, temperature and the presence of air in the UF-equipment. The effect of milk treatment on the plasminogen activation system should be further investigated. [Copyright &y& Elsevier]

Mammary involution and inflammation are known to negatively affect milk quality. A trial was carried out to elucidate the mechanism by which udder health status and lactational phase determine compositional modifications in ovine milk. A total of 60 individual milk samples was collected from a group of 20 pluriparous Sardinian ewes from mid to late lactation. Each sample was assessed for its chemical characteristics, quantitative distribution of casein fractions, lactodynamographic characteristics, and enzymatic activity. Udders were classed as healthy, doubtful, or infected on the basis of repeated somatic cell counts, and samples were grouped in 3 classes of days in milk. Results indicated that both udder inflammation and mammary involution can increase plasmin (PL) activity (15.6 vs. 18.4 U/mL in healthy vs. infected udders; 14.0 vs. 20.2 U/mL in phase 1 vs. 3), which is responsible for an evident protein breakdown in milk. Significant differences between groups were observed for several characteristics. With regard to udder heath status, casein index was lower in the infected vs. healthy udders (74.8 vs. 76.6%), and βtot-casein showed a similar trend (43.9 vs. 46.6%). As a consequence of protein degradation, γ-casein (5.78 vs. 2.82%) and proteolysis index (7.60 vs. 3.82) increased in the infected group with respect to the healthy group. Udder health status also affected milk technological traits. Udder inflammation resulted in longer clotting time (20.7 vs. 16.5 min for infected vs. healthy, respectively) and in poorer curd firmness (35.6 vs. 47.6 mm for infected vs. healthy, respectively). Frequency of samples reactive to rennet was 100, 93, and 67%, respectively, for healthy, doubtful, and infected groups. With regard to lactational phase, a decrease in αs1-casein (39.13 vs. 29.36%) and β1-casein (23.41 vs. 19.36%) occurred during phase I vs. 3, whereas κ + αs2... [ABSTRACT FROM AUTHOR]

The experiment was conducted from March to July 2002 using 5 intensively managed flocks of Southern Italy. In each flock, 2 groups of 50 ewes were created. The groups were designated LSCC (low somatic cell count [SCC]) when their milk SCC was lower than 500,000/mL and HSCC (high SCC) when their milk SCC was higher than 1,000,000/mL. Bulk milk and whey samples were analyzed for fat, total protein, lactose, casein, and whey protein contents. Renneting properties of milk were also determined. Moisture, NaCl, and nitrogen fractions were determined in fresh cheese curds. In addition, plasmin (PL) and plasminogen (PG) activities in milk and cheese were monitored. The proteolytic activity of plasmin by urea-polyacrylamide gel electrophoresis and the white blood cell (WBC) differentials were determined. The HSCC resulted in higher pH values in milk and in higher moisture and lower fat contents in fresh cheese curds. Moreover, a lower recovery of fat and whey proteins was obtained from the HSCC than from the LSCC raw milk. The crude protein and casein contents were higher in the HSCC than in the LSCC curds during early and midlactation; an opposite trend was observed in late lactation. Plasmin and PG activities underwent more marked fluctuations in the LSCC than in the HSCC curds through lactation. The results of this experiment demonstrate that the PL activity in ewe milk is markedly influenced by the SCC, although SCC is not the only parameter for predicting PL and PG evolution in ewe milk. The LSCC milk resulted in a higher proteolytic potential of Canestrato pugliese cheese curds. [ABSTRACT FROM AUTHOR]

Epistaxis is one of the most frequent hemorrhages resulting from local or systemic factors. Its management without hospitalization has prompted an interest in locally applied hemostatic agents. Generally, the therapy approaches involve sprays or creams acting as a physical barrier, even used as tampons or gauze. In this study, we have investigated the activity of Emoxilane®, a combination of sodium hyaluronate, silver salt, α-tocopherol acetate and D-panthenol, which is known to be able to separately act in a different biological manner. Our in vitro results, obtained on endothelial and nasal epithelial cells, have shown that the association of these molecules presented a notable antioxidant activity mainly due to the α-tocopherol and D-panthenol and a significant antimicrobial role thanks to the silver compound. Moreover, remarkable hemostatic activity was found by evaluating plasmin inhibition attributable to the sodium hyaluronate. Interestingly, on human plasma, we have confirmed that Emoxilane® strongly induced the increase of thrombin levels. These data suggest that the use of this association could represent an appealing pharmacological approach to actively induce hemostasis during epistaxis. Our future perspective will aim to the creation of a formulation for an easy topical application in the nose which is able to contrast the bleeding. [ABSTRACT FROM AUTHOR]

Zymographic and immunological studies revealed that primarily tissue-type plasminogen activator and to a lesser extent urokinase-type plasminogen activator were present in fluids of pemphigus vulgaris (type Neumann) skin blisters. Furthermore, plasmin activity was detected in pemphigus blister fluids using chromogenic peptide substrate assays. In pemphigus, but not in control, suction blister fluids plasmin/α-antiplasmin and plasmin/ α-macroglobulin complexes were found by immunoprecipitation or by testing in immunoassays after fractionation by molecular-sieve chromatography. Plasmin activity, detected by a low molecular weight chromogenic peptide assay, was ascribed to plasmin/α-macroglobulin complexes. Since formation of plasmin/inhibitor complexes requires active plasmin, the finding indicates previous activation of plasminogen in pemphigus lesions. [ABSTRACT FROM AUTHOR]