A Sensitive, Rapid, and Specific Technique for the Detection of Collagenase Using Zymography.

In-gel zymography is a commonly employed tool to identify active enzymes in a quantitative and qualitative manner. In this work, apart from the incorporation of substrate which is traditionally employed in zymography, the identification of collagenase by incubation of the enzyme resolved on a polyacrylamide gel with substrate solution is described. The two methods are quite fast and result in specific detection of bacterial collagenase.

Collagenase clostridium histolyticum in Dupuytren's contracture: a systematic review.

INTRODUCTION: In the last few years, the use of collagenase clostridium histolyticum for management of Dupuytren's contracture has increased. The procedure of enzymatic fasciectomy has become popular because it is non-invasive, safe and fast to perform. SOURCES OF DATA: A systematic search was performed on Medline (PubMed), Web of Science and Scopus databases using the combined keywords 'Dupuytren collagenase' and 'Dupuytren clostridium histolyticum'. Forty-three studies were identified. The quality of the studies was assessed using the Coleman Methodological Score. AREAS OF AGREEMENT: The use of collagenase clostridium histolyticum provides better outcomes in patients with mild-moderate joint contracture, with lower complications and side effects than open fasciectomy. Manipulation can be performed 2-7 days after the injection. The use of collagenase is cost-effective. AREAS OF CONTROVERSY: Most of the studies did not report patient-related outcomes. The role of dynamic splint has to be investigated with randomized clinical trials. GROWING POINTS: The shorter recovery time and the low incidence of serious or major adverse effects are the main advantages of this new technology. AREAS TIMELY FOR DEVELOPING RESEARCH: There is a need to perform studies with longer follow-up because the recurrence rate seems to increase with time. Further investigations are necessary to assess whether it is safe and effective to inject two or more cords at the same time.

The 32nd Annual J.P. Morgan Healthcare Conference (January 13-16, 2014 - San Francisco, California, USA): Auxilium Pharmaceuticals.

CEO and President Adrian Adams described Auxilium Pharmaceuticals as "the same as a Big Pharma company, only smaller." This 'specialty biopharmaceutical company' has transformed itself in the last 6 to 9 months to become the "leading men's healthcare franchise with a broad and diverse product portfolio." All of this has led to a very busy 2013, which began with the USD 600 million acquisition (Q2) and integration of Actient Pharmaceuticals, the licensing of avanafil (Stendra™) for erectile dysfunction and ended with the approval of collagenase Clostridium histolyticum (Xiaflex®) in Peyronie's disease. From a portfolio of 2 products at the beginning of 2013, Auxilium ended the year with 12.

Use of resources and costs associated with the treatment of Dupuytren's contracture at an orthopedics and traumatology surgery department in Denia (Spain): collagenase clostridium hystolyticum versus subtotal fasciectomy.

BACKGROUND: Our purpose was to analyze and compare the use of direct health resources and costs generated in the treatment of Dupuytren's contracture using two different techniques: subtotal fasciectomy and infiltration with Collagenase Clostridium Histolyticum (CCH) in regular clinical practice at the Orthopedic and Traumatology Surgery (OTS) Department at the Hospital de Denia (Spain). METHODS: Observational, retrospective study based on data from the computerized clinical histories of two groups of patients- those treated surgically using a one or two digit subtotal fasciectomy technique (FSC) and those treated with CCH infiltration, monitored in regular clinical practice from February, 2009 to May, 2012. Demographic (age, sex), clinical (number of digits affected and which ones) and use of resources (hospitalizations, medical visits, tests and drugs) data were collected. Resource use and associated costs, according to the hospital's accounting department, were compared based on the type of treatment from Spain's National Health Service. RESULTS: 91 patients (48 (52.8%) in the FSC group) were identified. The average age and number of digits affected was 65.9 (9.2) years and 1.33 (0.48) digits affected in the FSC group, and 65.1 (9.7) years and 1.16 (0.4) digits in the CCH group.Overall, the costs of treating Dupuytren's disease with subtotal FSC amount to €1,814 for major ambulatory surgery and €1,961 with hospital stay including admission, surgical intervention (€904), examinations, dressings and physiotherapy. As to collagenase infiltration, costs amount to €952 (including minor surgery admission, vial with product, office examination and dressings). Finally, comparing total costs for treatments, a savings of €388 is estimated in favor of CCH treatment in the best-case scenario (patient under MAS system with no need for physiotherapy) and €1,008 in the worst-case scenario (patient admitted to hospital needing subsequent physiotherapy), implying a savings of 29% and 51%, respectively. CONCLUSIONS: This study demonstrates that treating patients with DC by injection with CCH at the OTS department of the Hospital de Denia generates a total savings of 29% and 51% (€388 and €1008) compared with fasciectomy at the time of treatment. Long term evolution of CCH treatment is uncertain and the recurrence rate unknown.

Budget impact analysis in Spanish patients with Dupuytren's contracture: fasciectomy vs. collagenase Clostridium histolyticum.

The aim of this study was to estimate the budget impact of collagenase Clostridium histolyticum (CCH) vs. fasciectomy (FSC) surgery for the treatment of Dupuytren's disease (DD) in Spain. A cost minimization analysis was adopted (effectiveness was assumed to be equivalent for both techniques). DD related costs were considered. CCH costs (including drug, administration and visits) were obtained from clinical trials and a real-life study. FSC costs (including type of admission, visits, operating room, re-admissions, tests, drugs and rehabilitation costs) were collected through a retrospective, observational, local study. Unit costs were obtained from local database systems (e-SALUD and BOT). Results were presented from the NHS perspective for the next 3 years. We assumed that there were 5100 fasciectomies per year (with a 5% annual increase) and that 20%, 30% and 40% of them will annually utilize CCH. In addition, a 10%, 15% and 20% of untreated diagnosed patients were expected to receive CCH. All the data were validated through an expert panel. A sensitivity analysis was performed with the main variables. The average FSC cost was €2250 (72% inpatients), €1703 for outpatients and €2467 for inpatients. The average CCH cost was €1220 (1.5 vial/injection and four visits) and could drop to €898 (1.1 vial/injections and three visits). The accumulated 3years budget impact analysis (BIA) was 45,971€ (K€-2993(1); 3870). According to this study, the inclusion of the CCH should produce a 3-year cumulative budgetary impact of €45,971 (K€-2993; 3870) for the NHS.

▾Xiapex for Dupuytren's contracture.

Dupuytren's contracture is a benign, progressive disabling condition of the hands. Traditionally, surgery (e.g. fasciotomy, fasciectomy, joint fusion and amputation) has been the only effective treatment but it is not an option for every patient. A potential alternative intervention is a formulation of collagenase Clostridium histolyticum (▾Xiapex--Pfizer Limited), which has been licensed for injection directly into the affected joint as treatment for adults with Dupuytren's contracture.5 Here we review the evidence for collagenase C. histolyticum and discuss its place in the management of this condition.

Tissue dissociation enzyme neutral protease assessment.

Neutral proteases, essential components of purified tissue dissociation enzymes required for successful human islet isolation, show variable activities and effects of substrate on their activities. Initially we used a spectrophotometric endpoint assay with azocasein substrate to measure neutral protease activity. After critical review of the results, we observed these data to be inconsistent and not correlating expected differences in specific activities between thermolysin and Bacillus polymyxa proteases. This observation led to the development of a fluorescent microplate assay using fluorescein isothyocyanate-conjugated bovine serum albumin (FITC-BSA) as the substrate. This simpler, more flexible method offered a homogeneous, kinetic enzyme assay allowing determination of steady state reaction rates of sample replicates at various dilutions. The assay had a linear range of 4- to 8-fold and interassay coefficients of variation for B polymyxa protease and thermolysin of <9% and <15%, respectively, which were lower than those using the spectrophotometric endpoint assay, namely, 54% and 36%, respectively. This format allowed for incorporation of enzyme inhibitors, as illustrated by addition of sulfhydryl protease inhibitors, which, consistent with earlier reports, strongly indicated that the main contaminant in purified collagenase preparations was clostripain. Determination of the specific activities for several purified neutral proteases showed that the B polymyxa and Clostridium histolyticum proteases had approximately 40% and 15% specific activities, respectively, of those obtained with purified thermolysin, indicating the different characteristics of neutral protease enzymes for cell isolation procedures.