Rapid bioassay for evaluating enzyme production in fungal isolates from environmental sources

Fungal hydrolytic enzymes have a great potential due to the rapid development of enzyme technology and their industrial applications. A fluorimetric microplate assay has been developed for measuring activity of β-D-exoglucanase, β-D-glucosidase, α-D-glucosidase, β-N-acetyl-hexosaminidase, β-D-xylosidase in diluted culture broth samples. The substrates used are conjugates of the highly fluorescent compounds 4-methylumbelliferone (MUB) and thus product formation can be measured directly in the microplate without previous extraction and purification of the product. The developed system was evaluated on eight fungal strains isolated from shrub species (Tamarix) that can be used in phytoremediation. Depending upon the enzyme, each species exhibited different levels of enzymatic activities as well as different production profiles. The fluorimetric method could constitute an effective alternative to the pectrophotometric method to screen hydrolase-producing microbial strains

Evaluation of Clinical Outcomes After Revascularization in Patients With Chronic Limb-Threatening Ischemia: Results From a Prospective National Cohort Study (RIVALUTANDO)

We evaluated the outcomes of revascularization in patients with chronic limb-threatening ischemia (CLTI) treated in real-world settings. This is a prospective multicenter cohort study with 12-month follow-up enrolling patients (n = 287) with CLTI undergoing open, endovascular, or hybrid lower extremity revascularization. The primary end point was amputation-free survival (AFS) at 12 months. Cox proportional analysis was used to determine independent predictors of amputation and restenosis. At 30 days, major adverse cardiovascular and major adverse limb events (MALE) rates were 3.1% and 2.1%, respectively. At 1 year, the overall survival rate was 88.8%, the AFS was 86.6%, and the primary patency was 70.5%. Freedom from MALE was 62.5%. After multivariate analysis, smoking (hazard ratio [HR] = 2.2, P = 0.04), renal failure (HR = 2.3, P = 0.03), Rutherford class (≥5) (HR = 3.2, P = 0.01), and below-the-knee disease (HR = 2.0, P = 0.05) were significant predictors of amputation; iloprost infusion (>10 vials) (HR = 0.64, P = 0.05) was a significant protective factor. Cilostazol administration (HR = 0.77, P = 0.05) was a significant protective factor for restenosis. Results from this prospective multicenter registry offer a consistent overview of clinical outcomes of CLTI patients at 1 year when adequately revascularized. Medical treatment, including statins, cilostazol and iloprost, were associated with improved 1-year freedom from restenosis and amputation