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      The Chemical and Antibacterial Evaluation of St. John's Wort Oil Macerates Used in Kosovar Traditional Medicine

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          Abstract

          Hypericum perforatum L. (Hypericaceae), or St. John's Wort, is a well-known medicinal herb often associated with the treatment of anxiety and depression. Additionally, an oil macerate (Oleum Hyperici) of its flowering aerial parts is widely used in traditional medicine across the Balkans as a topical wound and ulcer salve. Other studies have shown that Oleum Hyperici reduces both wound size and healing time. Of its active constituents, the naphthodianthrone hypericin and phloroglucinol hyperforin are effective antibacterial compounds against various Gram-positive bacteria. However, hyperforin is unstable with light and heat, and thus should not be present in the light-aged oil macerate. Additionally, hypericin can cause phototoxic skin reactions if ingested or absorbed into the skin. Therefore, the established chemistry presents a paradox for this H. perforatum oil macerate: the hyperforin responsible for the antibacterial bioactivity should degrade in the sunlight as the traditional oil is prepared; alternately, if hypericin is present in established bioactive levels, then the oil macerate should cause photosensitivity, yet none is reported. In this research, various extracts of H. perforatum were compared to traditional oil macerates with regards to chemical composition and antibacterial activity (inhibition of growth, biofilm formation, and quorum sensing) vs. several strains of Staphylococcus aureus in order to better understand this traditional medicine. It was found that four Kosovar-crafted oil macerates were effective at inhibiting biofilm formation (MBIC 50 active range of 0.004–0.016% v/v), exhibited moderate inhibition of quorum sensing (QSIC 50 active range of 0.064–0.512% v/v), and contained detectable amounts of hyperforin, but not hypericin. Overall, levels of hypericin were much higher in the organic extracts, and these also exhibited more potent growth inhibitory activity. In conclusion, these data confirm that oil macerates employed in traditional treatments of skin infection lack the compound credited with phototoxic reactions in H. perforatum use and exhibit anti-biofilm and modest quorum quenching effects, rather than growth inhibitory properties against S. aureus.

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          Identification of Genes Involved in Polysaccharide-Independent Staphylococcus aureus Biofilm Formation

          Staphylococcus aureus is a potent biofilm former on host tissue and medical implants, and biofilm growth is a critical virulence determinant for chronic infections. Recent studies suggest that many clinical isolates form polysaccharide-independent biofilms. However, a systematic screen for defective mutants has not been performed to identify factors important for biofilm formation in these strains. We created a library of 14,880 mariner transposon mutants in a S. aureus strain that generates a proteinaceous and extracellular DNA based biofilm matrix. The library was screened for biofilm defects and 31 transposon mutants conferred a reproducible phenotype. In the pool, 16 mutants overproduced extracellular proteases and the protease inhibitor α2-macroglobulin restored biofilm capacity to 13 of these mutants. The other 15 mutants in the pool displayed normal protease levels and had defects in genes involved in autolysis, osmoregulation, or uncharacterized membrane proteins. Two transposon mutants of interest in the GraRS two-component system and a putative inositol monophosphatase were confirmed in a flow cell biofilm model, genetically complemented, and further verified in a community-associated methicillin-resistant S. aureus (CA-MRSA) isolate. Collectively, our screen for biofilm defective mutants identified novel loci involved in S. aureus biofilm formation and underscored the importance of extracellular protease activity and autolysis in biofilm development.
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            Mutation of sarA in Staphylococcus aureus limits biofilm formation.

            Mutation of sarA resulted in a reduced capacity to form a biofilm in six of the eight Staphylococcus aureus strains we tested (UAMS-1, UAMS-601, SA113, SC-01, S6C, and DB). The exceptions were Newman, which formed a poor biofilm under all conditions, and RN6390, which consistently formed a biofilm only after mutation of agr. Mutation of agr in other strains had little impact on biofilm formation. In every strain other than Newman, including RN6390, simultaneous mutation of sarA and agr resulted in a phenotype like that observed with the sarA mutants. Complementation studies using a sarA clone confirmed that the defect in biofilm formation was due to the sarA mutation.
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              Clinical use of Hypericum perforatum (St John's wort) in depression: A meta-analysis.

              St John's wort is a popular herbal remedy recommended by Traditional Chinese Medicine (TCM) practitioners and licensed and widely prescribed for depression in many European countries. However, conflicting data regarding its benefits and risks exist, and the last large meta-analysis on St John's wort use for depression was done in 2008, with no updated meta-analysis available.
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                Author and article information

                Contributors
                Journal
                Front Microbiol
                Front Microbiol
                Front. Microbiol.
                Frontiers in Microbiology
                Frontiers Media S.A.
                1664-302X
                08 September 2017
                2017
                : 8
                : 1639
                Affiliations
                [1] 1Center for the Study of Human Health, Emory University Atlanta, GA, United States
                [2] 2Department of Dermatology, Emory University School of Medicine Atlanta, GA, United States
                [3] 3Department of Biology, University of Pristina Prishtinë, Kosovo
                [4] 4Emory Antibiotic Resistance Center, Emory University Atlanta, GA, United States
                Author notes

                Edited by: Tzi Bun Ng, The Chinese University of Hong Kong, Hong Kong

                Reviewed by: Chien-Yi Chang, Guangzhou Women and Children Medical Center, China; Ben W. Greatrex, University of New England, Australia; Atte Von Wright, University of Eastern Finland, Finland; Catherine M. T. Sherwin, University of Utah, United States; Sunil D. Saroj, Symbiosis International University, India

                *Correspondence: Cassandra L. Quave cquave@ 123456emory.edu

                This article was submitted to Antimicrobials, Resistance and Chemotherapy, a section of the journal Frontiers in Microbiology

                †These authors have contributed equally to this work.

                Article
                10.3389/fmicb.2017.01639
                5596533
                07901b0a-dd83-41c8-90c3-7ea0c9194643
                Copyright © 2017 Lyles, Kim, Nelson, Bullard-Roberts, Hajdari, Mustafa and Quave.

                This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

                History
                : 01 March 2017
                : 14 August 2017
                Page count
                Figures: 8, Tables: 5, Equations: 0, References: 61, Pages: 19, Words: 10604
                Funding
                Funded by: National Institutes of Health 10.13039/100000002
                Award ID: R01 AT007052
                Funded by: U.S. Department of State 10.13039/100000194
                Award ID: S-KV-420-14GR-096
                Categories
                Microbiology
                Original Research

                Microbiology & Virology
                hypericum perforatum,staphylococcus aureus,traditional medicine,phototoxicity

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