Urequinona, a molecule from the root of <i>Pentalinon andrieuxii</i> Muell-Arg heals <i>Leishmania mexicana</i> ear’s infection in mice: This plant is widely used by the Mayan traditional medicine

AbstractIn this work we tested both the in vitro and in vivo anti-Leishmania mexicana activity of a molecule we originally identified in the root of Pentalinon andrieuxii Muell-Arg, a plant that is widely used in Mayan traditional medicine. The chemical name of this molecule is 24-methylcholesta-4-24(28)-dien-3-one and for simplicity we assigned the short and trivial name of urequinona that will be used throughout this work. It induces necrosis and apoptosis of promastigotes cultured in vitro and extensive ultrastructural damage of both promastigotes and amastigotes. It also induces production of Interleukin (IL)-2 and interferon (IFN)-γ by splenic cells from infected and urequinona treated mice stimulated in vitro with parasite antigen (Ag) but inhibits production of IL-6 and IL-12p70 by bone marrow derived macrophages (BMM) infected in vitro and then treated with urequinona. It also induces activation of transcription factors such as NFkB and AP-1 (NFkB/AP-1) in RAW reporter cells. We also developed a novel pharmaceutical preparation of urequinona encapsulated in hydroxyethyl cellulose for dermal application that significantly reduced experimentally induced ear′s lesions of C57BL/6 mice. We conclude the preparation containing this molecule is a good candidate for a novel anti-leishmanial drug′s preparation.</jats:p

Development of an automated image analysis protocol for quantification of intracellular forms of Leishmania spp.

Leishmania parasites cause a set of neglected tropical diseases with considerable public health impact, the leishmaniases, which are often fatal if left untreated. Since current treatments for the leishmaniases exhibit high toxicity, low efficacy and prohibitive prices, many laboratories throughout the world are engaged in research for the discovery of novel chemotherapeutics. This entails the necessity of screening large numbers of compounds against the clinically relevant form of the parasite, the obligatory intracellular amastigote, a procedure that in many laboratories is still carried out by manual inspection. To overcome this well-known bottleneck in Leishmania drug development, several studies have recently attempted to automate this process. Here we implemented an image-based high content triage assay for Leishmania which has the added advantages of using primary macrophages instead of macrophage cell lines and of enabling identification of active compounds against parasite species developing both in small individual phagolysosomes (such as L. infantum) and in large communal vacuoles (such as L. amazonensis). The automated image analysis protocol is made available for IN Cell Analyzer systems, and, importantly, also for the open-source CellProfiler software, in this way extending its implementation to any laboratory involved in drug development as well as in other aspects of Leishmania research requiring analysis of in vitro infected macrophages.Project Norte-01-0145-FEDER-000012Structured program on bioengineered therapies for infectiousdiseases and tissue regeneration, supportedby Norte Portugal Regional 17 Operational Programme (NORTE 2020), under the PORTUGAL 2020 PartnershipAgreement,through the European Regional Development Fund (FEDER). AGGA and TC were supported by contracts SFRH/BD/93766/2013,financed by POPH—QREN, and subsidized by Fundo Social Europeu and Ministe ´rio da Ciência,Tecnologia e Ensino Superior, and PTDC/QEQ-MED/7097/2014, funded by National Funds throughFCT, respectively. HC is fundedby FCT under the “Investigador FCT” Programme.info:eu-repo/semantics/publishedVersio