Optimization of production, biochemical characterization and In Vitro evaluation of the therapeutic potential of fibrinolytic enzymes from a new Bacillus Amyloliquefaciens

The capacity of fibrinolytic enzymes to degrade blood clots makes them of high relevance in medicine and in the pharmaceutical industry. In this work, forty-three microorganisms of the genus Bacillus were evaluated for their potential to produce fibrinolytic proteases. Thirty bacteria were confirmed as producers of fibrinolytic enzymes, the best results obtained for the strain Bacillus amyloliquefaciens UFPEDA 485. The optimization of the enzyme production conditions was done by a central composite design (CCD) star 23 that allowed to define the optimal conditions for soybean flour and glucose concentrations and agitation rate. The highest fibrinolytic activity (FA) of 813 U mL-1 and a degradation of blood clot in vitro of 62% were obtained in a medium with 2% (w/v) of soybean flour and 1% (w/v) glucose at 200 rpm after 48 h of cultivation, at pH 7.2 and 37 °C. The obtained fibrinolytic enzyme was characterized biochemically. Fibrinolytic activity was inhibited by PMSF (fluoride methylphenylsulfonyl - C7H7FO2S) 91.52% and EDTA (ethylenediaminetetraacetic acid - C10H16N2O8) 89.4%, confirming to be a serine- metallo protease. The optimum pH and temperature were 7.0 and 37 oC, respectively, and the enzyme was stable for 12 h. The fibrinolytic activity at physiological conditions of this enzyme produced by Bacillus amyloliquefaciens UFPEDA 485, as well as its long term stability, demonstrate that it has suitable characteristics for human and veterinary applications, and promises to be a powerful drug for the treatment of vascular diseases.We express our thanks to Coordination for the Improvement of Higher Level Education Personnel (CAPES) - Doctoral Sandwich Program (PDSE) Nº 0259/ 12-8 and National Council for Scientific and Technological Development (CNPq) - Nº 202026/2011-6 for the financial support

Neuroprotective impact of Ximenia americana aqueous bark extract on Diazepam-induced memory impairment in mice via its antioxidant potential

In traditional medicine, Ximenia americana (XA) is used to treat mental disorders, and headaches. The current study aimed to show the preventive and biochemical impacts of XA aqueou’s extract on diazepam-induced amnesia. Mice were randomized as follows: distilled water (10 mL/kg); diazepam (3 mg/kg); piracetam (PIR) (150 mg/kg); and XA experimental groups (25, 50 and 75 mg/kg). Mice were then treated in groups, 14 straight days. Radial arm maze (RAM) and T-maze were employed to assess different behaviours 30 min after each treatment. After the test was completed, the brains were isolated for histological and biochemical examinations. The results obtained showed that XA extract seriously (p < 0.001) reversed mistakes in working remembrance in the radial arm maze test contrasted to the normal control factions. In the T-maze test, pretreatment of mice with XA extract seriously (p < 0.001) expanded the time spent in the preferred arm when contrasted to the DZP-only treated faction. The XA-treated DZP groups showed subsequent (p < 0.001) improvement in catalase (CAT) and reduced glutathione (GHS). A diminish in malondialdehyde (MDA) level was observed in brain homogenates of mice treated with the extract contrasted with the DZP- group. These few results regarding the neuroprotective and antioxidant effects of XA extract at least partially demonstrate its empirical use in the treatment of certain pathologies

Abstracts from the 20th International Symposium on Signal Transduction at the Blood-Brain Barriers

https://deepblue.lib.umich.edu/bitstream/2027.42/138963/1/12987_2017_Article_71.pd

Determination of taurine in soft drinks by an ultrahigh-performance liquid chromatography-mass spectrometry method

Taurine (2-aminoethanesulfonic acid) is a free sulfur-containing β-amino acid widely distributed in many mammalians. Owing to the energizing effects, it is mostly used in soft drinks and supplements for athletes. Regular intake of soft drinks may lead to an overdose of caffeine, taurine, and guarana and loss of bone mass, overweight, hypertension, and in older age, osteoporosis and cardiovascular diseases. Therefore, it is essential to control the maximum amount of taurine consumed by humans in the food and beverages. Here, a fast, simple, accurate, and robust method based on ultrahigh-performance liquid chromatography hyphenated with mass spectrometry (UHPLC-MS) was successfully applied for the determination of taurine in selected soft drinks sold in Slovakia. The method was characterized by coefficient of determination higher than 0.99, and the predicted value of the limit of detection was 4.29 μmol/L. The analyzed levels of taurine in selected commercial drinks ranged from 2.8 to 3.78 mg/mL. The concentration in one brand of the investigated drinks was found to be extremely low (about 70%) compared to the declared content by the manufacturer