PMID- 27036961 OWN - NLM STAT- MEDLINE DCOM- 20161101 LR - 20201209 IS - 1472-6882 (Electronic) IS - 1472-6882 (Linking) VI - 16 DP - 2016 Mar 31 TI - Exploration of anti-Malassezia potential of Nyctanthes arbor-tristis L. and their application to combat the infection caused by Mala s1 a novel allergen. PG - 114 LID - 10.1186/s12906-016-1092-2 [doi] LID - 114 AB - BACKGROUND: Malassezia commensal yeasts along with multitude of antigens have been found to be associated with various skin disorders including Pityriasis versicolor (PV). Amongst them Mala s1, a 37 kDa protein has been proved to be a major allergen reacting with a large panel of sera. However, there exists no therapeutic alternative to combat such problems in form of plant based natural compounds. The purpose of this study is in the first place, to determine the anti-Malassezia activity of Nyctanthes arbor-tristis L. (NAT) ethanolic leaf extract through turbidimetric growth curves, disruption of plasma membrane and secondly, it aims to present in silico validation of its active constituents over Mala s1a novel allergen. METHODS: The antifungal susceptibility 50 % ethanolic extract of NAT was determined by broth microdilution method according to CLSI guidelines. Further MICs and IC50 were determined spectrophotometrically using the software SoftMax(R) Pro-5 (Molecular Devices, USA). Active constituents mediated disruption of plasma membrane was studied through flowcytometry by permeabilization of fluorescent dye Propidium Iodide (PI). Antioxidant activity of the extract was determined using the DPPH stable radical. Molecular validation of fungal DNA from the extract was observed using PCR amplification. In silico analysis of its active constituents over Mala s1 was performed using HEX software and visualized through Pymol. RESULTS: The anti-Malassezia potential of NAT leaf extracts reflected moderate MIC 1.05 mug/mul against M. globosa, while least effective against M. restricta with MIC 1.47 mug/mul. A linear correlation coefficient R (2) = 0.866 was obtained in case of M. globosa while minimum was observed in M. restricta with R (2) = 0.732. The flow cytometric data reveal ~ 75 % cell death when treated with active constituents beta-Sitosterol and Calceolarioside A. The docking confirmations and the interaction energies between Mala s1 and the active constituents (beta-Sitosterol and Calceolarioside A) from extracts showed an effective binding which suggests Mala s1 as efficient allergen for site specific targeting. CONCLUSIONS: This study revealed that Nyctanthes arbor-tristis L. (NAT) extracts possess high anti-Malassezia potential which is driven mainly by disruption of plasma membrane. Also in silico validation and molecular modeling studies establishes Mala s1 as a novel allergen that could be a potential target in disease treatment. Our results would also provide a foundation for the development of new therapeutic approach using NAT extract as lead compound with high antioxidant property as an added trait for skin care. FAU - Mishra, Rohit K AU - Mishra RK AD - Centre for Medical Diagnostic and Research (CMDR), Motilal Nehru National Institute of Technology (MNNIT), Allahabad, 211004, India. rohit_ernet@yahoo.co.in. FAU - Mishra, Vani AU - Mishra V AD - Nanotechnology Application Centre (NAC), University of Allahabad, Allahabad, 211002, India. vanimish@gmail.com. FAU - Pandey, Anand AU - Pandey A AD - Department of Botany, Biological Product Lab., University of Allahabad, Allahabad, 211002, India. FAU - Tiwari, Amit K AU - Tiwari AK AD - Department of Botany, Biological Product Lab., University of Allahabad, Allahabad, 211002, India. FAU - Pandey, Himanshu AU - Pandey H AD - Department of Pharmaceutical Sciences, Sam Higginbottom Institute of Agriculture Technology and Sciences, Allahabad, 211007, India. FAU - Sharma, Shivesh AU - Sharma S AD - Centre for Medical Diagnostic and Research (CMDR), Motilal Nehru National Institute of Technology (MNNIT), Allahabad, 211004, India. FAU - Pandey, Avinash C AU - Pandey AC AD - Nanotechnology Application Centre (NAC), University of Allahabad, Allahabad, 211002, India. FAU - Dikshit, Anupam AU - Dikshit A AD - Department of Botany, Biological Product Lab., University of Allahabad, Allahabad, 211002, India. anupambplau@rediffmail.com. LA - eng PT - Journal Article PT - Research Support, Non-U.S. Gov't DEP - 20160331 PL - England TA - BMC Complement Altern Med JT - BMC complementary and alternative medicine JID - 101088661 RN - 0 (Allergens) RN - 0 (Antifungal Agents) RN - 0 (Antigens, Fungal) RN - 0 (Antioxidants) RN - 0 (Caffeic Acids) RN - 0 (Glucosides) RN - 0 (Mala s 1 allergen, Malassezia sympodialis) RN - 0 (Plant Extracts) RN - 0 (Sitosterols) RN - 5LI01C78DD (gamma-sitosterol) RN - 84744-28-5 (calceolarioside A) SB - IM MH - Allergens/*immunology/metabolism MH - Antifungal Agents/metabolism/*therapeutic use MH - Antigens, Fungal/*immunology/metabolism MH - Antioxidants/therapeutic use MH - Caffeic Acids/metabolism MH - Cell Membrane/drug effects MH - Dermatomycoses/*drug therapy MH - Glucosides/metabolism MH - India MH - Malassezia/*immunology MH - Microbial Sensitivity Tests MH - Oleaceae/*chemistry MH - Plant Extracts/metabolism/*therapeutic use MH - Sitosterols/metabolism PMC - PMC4815214 OTO - NOTNLM OT - AUGC OT - Anti-Malassezia susceptibility OT - Flow cytometry OT - Mala s1 OT - Molecular docking OT - Nyctanthes arbor-tristis L. (NAT) EDAT- 2016/04/03 06:00 MHDA- 2016/11/02 06:00 PMCR- 2016/03/31 CRDT- 2016/04/03 06:00 PHST- 2015/05/02 00:00 [received] PHST- 2016/03/23 00:00 [accepted] PHST- 2016/04/03 06:00 [entrez] PHST- 2016/04/03 06:00 [pubmed] PHST- 2016/11/02 06:00 [medline] PHST- 2016/03/31 00:00 [pmc-release] AID - 10.1186/s12906-016-1092-2 [pii] AID - 1092 [pii] AID - 10.1186/s12906-016-1092-2 [doi] PST - epublish SO - BMC Complement Altern Med. 2016 Mar 31;16:114. doi: 10.1186/s12906-016-1092-2.