Characterization of different alginate lyases for dissolving Pseudomonas aeruginosa biofilms.

Aggregates of Pseudomonas aeruginosa form a protective barrier against antibiotics and the immune system. These barriers, known as biofilms, are associated with several infectious diseases. One of the main components of these biofilms is alginate, a homo- and hetero-polysaccharide that consists of ß-D-mannuronate (M) and α-L-guluronate (G) units. Alginate lyases degrade this sugar and have been proposed as biotherapeutic agents to dissolve P. aeruginosa biofilms. However, there are contradictory reports in the literature regarding the efficacy of alginate lyases against biofilms and their synergistic effect with antibiotics. We found that most positive reports used a commercial crude extract from Flavobacterium multivorum as the alginate lyase source. By using anion exchange chromatography coupled to nano LC MS/MS, we identified two distinct enzymes in this extract, one has both polyM and polyG (polyM/G) degradation activities and it is similar in sequence to a broad-spectrum alginate lyase from Flavobacterium sp. S20 (Alg2A). The other enzyme has only polyG activity and it is similar in sequence to AlyA1 from Zobellia galactanivorans. By characterizing both of these enzymes together with three recombinant alginate lyases (a polyM, a polyG and a polyM/G), we showed that only enzymes with polyM/G activity such as Alg2A and A1-II' (alginate lyase from Sphingomonas sp.) are effective in dissolving biofilms. Furthermore, both activities are required to have a synergistic effect with antibiotics.

Condoliase for treatment of lumbar disc herniation.

Lumbar disc herniation (LDH) is generally treated with a conservative therapy, and surgery is the only therapeutic option currently available for patients unresponsive to the conservative therapy. In the 1980s, chemonucleolysis with chymopapain, a protease, was widely used as the intermediate treatment between conservative therapy and surgical therapy in the Western countries. However, since chymopapain was withdrawn from the market in 2002 for non-scientific commercial reasons, chemonucleolysis has not been a therapeutic option for LDH. Condoliase (chondroitin sulfate ABC endolyase), a glycosaminoglycan-degrading enzyme, was approved by the drug regulatory authority in Japan as a newer intradiscal therapy for LDH after clinical studies conducted in Japan demonstrated efficacy and safety for patients with LDH. This review will focus on the preclinical pharmacology, pharmacokinetics, efficacy and safety of condoliase as a new option for treatment of LDH.

Condoliase for the Treatment of Lumbar Disc Herniation: A Randomized Controlled Trial.

STUDY DESIGN: A randomized, double-blind, placebo-controlled, multicenter phase III clinical trial. OBJECTIVE: To evaluate the efficacy and safety of chemonucleolysis with condoliase in patients with lumbar disc herniation (LDH). SUMMARY OF BACKGROUND DATA: Condoliase is a pure mucopolysaccharidase derived from a bacterium, Proteus vulgaris that has high substrate specificity for chondroitin sulfate and hyaluronic acid in the nucleus pulposus of the intervertebral disc. METHODS: In this study, patients aged 20 to 70 years with unilateral leg pain, positive straight leg raise test, and a contained LDH were recruited in Japan. Patients were treated with a single injection of condoliase (1.25 U) or placebo and were followed for 1 year after administration. The primary endpoint was change in worst leg pain from baseline to week 13. The secondary endpoints included responder rate, and the changes from baseline up to week 52 in the worst leg pain, worst back pain, Oswestry Disability Index, 36-Item Short-Form Health Survey, neurologic examinations, and imaging parameters. RESULTS: A total of 82 and 81 patients received an injection of condoliase and placebo, respectively. The average changes in worst leg pain from baseline to week 13 (primary endpoint) were -49.5 mm in the condoliase group and -34.3 mm in the placebo group, and the difference of -15.2 mm was significant (95% confidence interval, -24.2 to -6.2; P = 0.001). Significant improvements were observed in the condoliase groups, compared with the placebo group, in most secondary endpoints at 1 year after administration. In the condoliase group, back pain, Modic type 1 change, and decrease in disc height were frequently reported, without any clinically relevant consequences. CONCLUSION: Condoliase significantly improved symptoms in patients with LDH and was well tolerated. Condoliase is a novel and potent chemonucleolytic drug for the treatment of LDH. LEVEL OF EVIDENCE: 1.

Phage in the diagnosis and treatment of tuberculosis.

The serious global TB epidemic coupled with limited diagnostic and therapeutic technologies necessitate the study of the role phage in TB treatment. Mycobacterium phage have been used for TB diagnosis, but the accuracy of such methods needs to be improved. Phage have various advantages in treating many kinds of bacterial infection, and coupled with the abuse and misuse of antibiotics, and the increasing prevalence of drug-resistant bacteria, they have been studied as a novel therapy to support antibiotics. The study of phage in TB therapy has developed from the selection of appropriate phage to the simultaneous use of multiple phage and even the use of purified lyase proteins. Though phage have great potential in TB therapy, the technology is still in the in vitro and animal experiment stages, and needs further study.

[Scleromyxedema with laryngeal changes. Beneficial results of the treatment with mucopolysaccharidases]. / Scléromyxoedème avec modifications laryngées. Résultats bénéfiques du traitement par mucopolysaccharidases.

We observed a case of scleromyxoedema with specific cutaneous lesions in a 62-year-old man. These lesions were associated with modifications of the central nervous system, latent thyroid failure, cardiovascular changes and myopathy, usual manifestations of this disease, but without paraproteinaemia. Unusual laryngeal manifestations were a dysphonia with chronic pseudomyxomatosis of the larynx. Urinary mucopolysaccharide excretion was increased. It is important to underscore the favourable response to Thiomucase when more toxic drugs had become ineffective.

[New therapy for Peyronie's disease: superoxide dismutase by ionization. Comparison with an earlier classical series]. / Nouveau traitement de la maladie de La Peyronie: la superoxyde dismutase par ionisations. Comparaison avec une série ancienne classique.

Due to the poor results obtained in the treatment of Peyronie's disease by ionophoresis and ultrasound, using the classical buffer solution of the cortisone thiomucase type, the authors have experimented ionophoresis with a new product: orgoteine a dismutase superoxide. Austrian authors have already demonstrated the value of this product. Its mechanism of action is still uncertain but it has an experimentally well demonstrated antiinflammatory activity. The good results are obvious not only on the pain, but also on the induration and incurvation, permitting the resumption of sexual intercourse in more than 75% of cases.

Influence of L-tyrosine phenol-lyase on the growth and metabolism of B16 melanoma.

Administration of L-tyrosine phenol-lyase inhibited growth of established B16 melanomas in BDF1 mice. Under the conditions used, the enzyme exerted a cytostatic effect on the tumors. The addition of L-tyrosine phenol-lyase to growing melanoma cultures blocked cell division and caused an accumulation of cells in the G1 (G0) pultures was suppressed, the most pronounced effect of L-tyrosine phenol-lyase was a rapid and near-complete cessation of uridine uptake and incorporation. This block in RNA synthesis was also observed in human fibroblasts treated with L-tyrosine phenol-lyase.

Some biological properties and an in vivo evaluation of tyrosine phenol-lyase on growth of B-16 melanoma.

Tyrosine phenol-lyase from Erwinia herbicola was purified with the goal of assessing its effect on growth of malignant melanoma. Ammonium sulfate-sodium citrate fractionation and diethylaminoethyl cellulose-hydroxylapatite chromatography were used. The purified enzyme was shown to reduce plasma tyrosine levels when administered to normal C57BL x DBA/2 F1 mice. The plasma half-life value of the enzyme was found to be 6 to 7 hr. Unlike results reported with glutaminase and asparaginase preparations, the lactate dehydrogenase-elevating virus had no significant influence on plasma clearance of tyrosine phenol-lyase. The enzyme significantly inhibited growth of established B-16 melanoma tumors.