The activity of polyphenolic compounds, triterpenoids, carotenoids, chlorophylls and antioxidants in leaves and rhizomes of Fallopia japonica Houtt and Fallopia sachalinensis (F.Schmidt) grown in Poland was investigated. Leaves and rhizomes were assessed for the presence of bioactive compounds with the ultra-performance liquid chromatography photodiode detector-quadrupole/time-of-flight mass spectrometry (UPLC-PDA-Q/TOF-MS) method, and for antioxidant activity with the on-line UPLC-ABTS screening. Forty-six polyphenolic compounds (15 phenolic acids, 12 flavones and flavonols, 11 flavan-3-ols and 8 stilbenes), were identified in Fallopia japonica and Fallopia sachalinensis. Furthermore, accurate mass measurement technique was for the first time in Fallopia japonica Houtt and Fallopia sachalinensis (F.Schmidt) in leaves and rhizomes it identified 25 new compounds belonging to carotenoids (9), chlorophylls (13) and triterpenoids (3) as well as rated the antioxidant properties of each polyphenolic compound. Major qualitative differences were found in the profiles. The leaves and rhizomes were found to be a good source not only of (average 20408.18 and 2716.42 mg/100 g dm), but also chlorophylls (average 179.97 and 43.82 mg/100 g dm), carotenoids (average 100.23 and 53.25 mg/100 g dm) and triterpenoids (average 580.87 and 434.05 mg/100 g dm). The content of bioactive compounds in Fallopia japonica Houtt was around 8.0, 4.0, 2.0 and 1.3 times higher than the content of polyphenols, chlorophylls, carotenoids and triterpenoids in Fallopia sachalinensis (F.Schmidt). The accurate identification of Fallopia bioactive compounds is an indispensable detailed knowledge of the profile and step toward better understanding of the medicinal properties of the species and also potentially more extensive use of the plant.
The cysteine protease inhibitor cystatin was purified from chicken egg white and its antimicrobial activity determined for a series of pathogenic bacteria. The results indicate that Acinetobacter lwoffii, Escherichia coli, Oligella sp. and Pseudomonas aeruginosa are highly sensitive to low concentrations of cystatin, which possesses bactericidal activity. No inhibition was observed with a Citrobacter freundii strain. Fifty percent growth inhibition (IC 50 ) was observed at cystatin concentrations in the range of 80 and 100 lg/ml, and the growth was completely inhibited at concentrations in the range of 100 and 200 lg/ml. Fifty percent growth inhibition (IC 50 ) for Staphylococcus aureus, Staphylococcus gallinarum, and Staphylococcus xylosus strains was observed at 150 and 200 lg of cystatin/ml respectively, and growth was completely inhibited at cystatin concentrations in the range of 300 and 1000 lg/ml. The activity of cysteine proteases was significantly decreased in the culture supernatant of microorganisms when chicken cystatin was added. In this study, we observed that chicken cystatin may be a candidate for antibacterial drug development aiming at controlling bacterial pathogens including Escherichia coli, Pseudomonas aeruginosa, and another possible application might be as a therapeutic agent for health improvement.
Our experiment demonstrated that the cysteine peptidases cathepsins B and L may be useful for the early detection of gastric cancer. The results suggest that addition of egg white cystatin reduces the activities of cathepsins B and L to that of non-cancerous values.
Cysteine cathepsin B and its endogenous inhibitor play an important role in tumor progression. Increase in cathepsin B expression and reduced levels of its inhibitors were associated with tumor malignancy in breast cancer. The objective of this study was to investigate the effects of a new therapy combining vitamin E and placental inhibitor on the level of endogenous protease inhibitor in sera and tumor tissues with mammary cancer. The inhibitor was used in doses of 100 and 200 micrograms per animal for 8 days. Vitamin E was added after the last treatment with inhibitor and was injected daily in doses of 10 and 20 mg per animal for one mouth. The size and survival time of treated animals as well as cathepsin B and the inhibitor activity in tumor and sera before and after treatment in comparison with the control groups were determined. The activity of cathepsin B significantly decreased both in tumor tissues and in sera (P < or = 0.0001). Cathepsin B activity in tumor tissue homogenates and in sera decreased two-fold and three-fold, respectively, after the animals were treated with vitamin E at a dose of 20 mg, and decreased five-fold and 15-fold, respectively, when treated with vitamin E plus inhibitor in comparison with untreated animals. Endogenous inhibitor activity increased six-fold and 12-fold in the sera and tissue homogenates, respectively, after the animals were treated with 200 micrograms of cysteine protease inhibitor plus 20 mg of vitamin E, in comparison with untreated animals. The total cure responses were higher in eight of 10 rats, as compared with untreated animals. The combination of placental inhibitor and vitamin E resulted in a significant reduction in breast metastasis and might provide a therapeutic basis for anti-metastasis therapy.
5-Aminolevulinic acid (5-ALA) is a well characterized precursor in the synthesis of various endogenous porphyrins used in photodynamic therapy (PDT). It is most often administered topically into a tumor which is then irradiated with visible light at established wavelength to sensitize porphyrins accumulated therein. Our main aim in the present study was to increase the penetration of 5-ALA through the altered skin by application of 3% azone (1-dodecyl-azepan-2-one) before the application of 20% 5-ALA in patients with plantar warts: mosaic warts (MW) and myrmecia (MY). We also used 20% 5-ALA only to treat warts in other patients. We compared the therapeutic and cosmetic effects of the two treatment modalities. The lesions treated with modification of 5-ALA-PDT by pretreatment with azone responded with better effectiveness. In 18 patients subjected to 5-ALA-PDT plus 3% azone, we observed 66.7% complete response of MW and 100% of MY following PDT repeated two or three times; whereas in other 18 patients treated with 5-ALA-PDT alone, we observed only 37.5% complete response of MW and 70% of MY. These results provide evidence that the pretreatment with azone should be considered as the step that enhances 5-ALA penetration in tissues and thus increases the effectiveness of applied PDT.
It was demonstrated that phenylmethanesulfonyl fluoride -a very potent inhibitor of penicillin amidase from Escherichia coli -binds covalently to the enzyme in molar ratio 1:1. The chloride, the azide and the Nhydroxysuccinimide ester of phenylmethanesulfonic acid are also very strong inactivators of the amidase. Weaker inhibition was noted with p0ra-substituted phenylmethanesulfonyl chlorides and with phenylethanesulfonyl and alkylsulfonyl chlorides. The inactivated amidase could be reactived by incubation either with 6-aminopenicillanic acid or with proteins from E. coli extract. Benzyl isocyanate is also a potent covalent inhibitor of the amidase but inactivated amidase could be not reactivated in this way. It was demonstrated that representatives of all inactivator types bind to one active site of the amidase. Interdependence between inactivation rate and stability of some sulfonyl inhibitors was observed. No inhibition was noted the amide, the hydrazide and the methyl ester of phenylmethanesulfonic acid. Phenylalkyhulfonyl-Derivate als kovalente Inhibitoren der Penicillin-ArnidaseZusammenfassung: Es wird gezeigt, daß Phenylmethansulfonylfluorid, ein sehr wirksamer Hemmstoff der Penicillin-Amidase aus Escherichia coli, im molaren Verhältnis l: l an das Enzym gebunden wird. Auch das Chlorid, das Azid und der Af-Hydroxysuccinimidester inaktivieren die Amidase weitgehend. Schwächere Hemmwerte wurden mit putra-substituierten Derivaten des Phenylmethansulfonylchlorids, mit Phenylethansulfonylchlorid und Alkylsulfonylchlorid gefunden. Die inaktivierte Amidase läßt sich durch Inkubation entweder mit 6-Aminopenicillansäure oder mit Proteinen aus einem E.-coliExtrakt reaktivieren. Auch Benzylisocyanat ist ein starker kovalenter Inhibitor der Amidase; das so inaktivierte Enzym ist aber nicht wie oben beschrieben reaktivierbar. Vertreter aller Inaktivatortypen werden an ein
Early detection and reliable monitoring of treatment constitute one of the most challenging problems in oncology. For this reason, we studied the potential value of serum cathepsin B-like activity as a possible marker of laryngeal carcinoma. The results indicate that serial analyses of this parameter have predictive value in the assessment of surgical treatment of laryngeal carcinoma. Activity of the enzyme reveals a progressive increment which could be associated with increased severity of the neoplasm present. The serum level of the cathepsin B-like activity remained close to zero in the control group of healthy volunteers and patients with inflammatory diseases.
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