The Discovery of Actinospene, a New Polyene Macrolide with Broad Activity against Plant Fungal Pathogens and Pathogenic Yeasts

Phytopathogenic fungi infect crops, presenting a worldwide threat to agriculture. Polyene macrolides are one of the most effective antifungal agents applied in human therapy and crop protection. In this study, we found a cryptic polyene biosynthetic gene cluster in Actinokineospora spheciospongiae by genome mining. Then, this gene cluster was activated via varying fermentation conditions, leading to the discovery of new polyene actinospene (1), which was subsequently isolated and its structure determined through spectroscopic techniques including UV, HR-MS, and NMR. The absolute configuration was confirmed by comparing the calculated and experimental electronic circular dichroism (ECD) spectra. Unlike known polyene macrolides, actinospene (1) demonstrated more versatile post-assembling decorations including two epoxide groups and an unusual isobutenyl side chain. In bioassays, actinospene (1) showed a broad spectrum of antifungal activity against several plant fungal pathogens as well as pathogenic yeasts with minimum inhibitory concentrations ranging between 2 and 10 μg/mL.

Dogs as Therapeutic Partners, Not Therapeutic Tools: Ethical Considerations for AAT in the Correctional Setting

Although the benefits of animal assisted therapy for people are well established, the ethical considerations for the welfare and safety of the non-human animals involved are not. Without an accrediting body responsible for creating and overseeing national standards, therapy animal organizations are forced to create their own guidelines, creating inconsistencies within the field. Based on interviews conducted with therapy teams who have worked with Parenting, Prison & Pups (PPP), a parenting program provided to incarcerated jailed women that is integrated with the use of animal-assisted therapy (AAT), this article explores the extent of ethics training offered for AAT teams and will examine how agencies and handlers promote and ensure the safety of canine partners, especially in a correctional setting. The research suggests that specific protocols put forth by individual AAT organizations, which can provide for a national model, can afford for the safety and comfortability of canine partners, especially in a corrections environment, but implies that in order to maintain consistency and increase therapy team professionalism, national standards are a necessity. Guidelines are specifically essential for mental health professionals, who lack guidelines from their own accrediting bodies’ code of ethics, and may incorporate non-human therapy partners into their work settings, without proper supervision.

Use of Exogenous Enzymes in Human Therapy: Approved Drugs and Potential Applications

: The development of safe and efficacious enzyme-based human therapies has increased greatly in the last decades, thanks to remarkable advances in the understanding of the molecular mechanisms responsible for different diseases, and the characterization of the catalytic activity of relevant exogenous enzymes that may play a remedial effect in the treatment of such pathologies. Several enzyme-based biotherapeutics have been approved by FDA (the U.S. Food and Drug Administration) and EMA (the European Medicines Agency) and many are undergoing clinical trials. Apart from enzyme replacement therapy in human genetic diseases, which is not discussed in this review, approved enzymes for human therapy find applications in several fields, from cancer therapy to thrombolysis and the treatment, e.g., of clotting disorders, cystic fibrosis, lactose intolerance and collagen-based disorders. The majority of therapeutic enzymes are of microbial origin, the most convenient source due to fast, simple and cost-effective production and manipulation. The use of microbial recombinant enzymes has broadened prospects for human therapy but some hurdles such as high immunogenicity, protein instability, short half-life and low substrate affinity, still need to be tackled. Alternative sources of enzymes, with reduced side effects and improved activity, as well as genetic modification of the enzymes and novel delivery systems are constantly searched. Chemical modification strategies, targeted- and/or nanocarrier-mediated delivery, directed evolution and site-specific mutagenesis, fusion proteins generated by genetic manipulation are the most explored tools to reduce toxicity and improve bioavailability and cellular targeting. This review provides a description of exogenous enzymes that are presently employed for the therapeutic management of human diseases with their current FDA/EMA-approved status, along with those already experimented at the clinical level and potential promising candidates.

Stromal Vascular Fraction: Biology and Application Outlook

Stromal vascular fraction (SVF) is a heterogeneous cell extract obtained with enzymatic dissociation of adipose tissue followed by centrifugation. This population includes many different cell types, i.a. adipose tissue stem cells (ATSCs), vascular endothelial and smooth muscle cells and their precursors, pericytes, fibroblasts, macrophages, T-lymphocytes, etc., excluding mature adipocytes. The main SVF component is ATSCs capable of self-renewal and multipotent differentiation. Since early research on SVF, an extensive effort has been aimed at understanding its clinical applications promoting a significant progress in the SVF use for treatment of various diseases and injuries. The past decade has witnessed an upward publication trend in basic and clinical research into the SVF therapeutic value. Manifold methods and devices for the SVF isolation from human liposuction lipoaspirate have been developed, continuously contributing to preclinical and clinical trials of its safety and efficacy. This review discusses the main properties and functions of the SVF cell population, its efficacy and safety for human therapy.

Methylene Blue Bridges the Inhibition and Produces Unusual Respiratory Changes in Complex III-Inhibited Mitochondria. Studies on Rats, Mice and Guinea Pigs

Methylene blue (MB) is used in human therapy in various pathological conditions. Its effects in neurodegenerative disease models are promising. MB acts on multiple cellular targets and mechanisms, but many of its potential beneficial effects are ascribed to be mitochondrial. According to the “alternative electron transport” hypothesis, MB is capable of donating electrons to cytochrome c bypassing complex I and III. As a consequence of this, the deleterious effects of the inhibitors of complex I and III can be ameliorated by MB. Recently, the beneficial effects of MB exerted on complex III-inhibited mitochondria were debated. In the present contribution, several pieces of evidence are provided towards that MB is able to reduce cytochrome c and improve bioenergetic parameters, like respiration and membrane potential, in mitochondria treated with complex III inhibitors, either antimycin or myxothiazol. These conclusions were drawn from measurements for mitochondrial oxygen consumption, membrane potential, NAD(P)H steady state, MB uptake and MB-cytochrome c oxidoreduction. In the presence of MB and complex III inhibitors, unusual respiratory reactions, like decreased oxygen consumption as a response to ADP addition as well as stimulation of respiration upon administration of inhibitors of ATP synthase or ANT, were observed. Qualitatively identical results were obtained in three rodent species. The actual metabolic status of mitochondria is well reflected in the distribution of MB amongst various compartments of this organelle.