Juliane Fernandes

University of São Paulo | USP · Ribeirão Preto School of Medicine (FMR

The identification of RNAs that are not translated into proteins was an important breakthrough, defining the diversity of molecules involved in eukaryotic regulation of gene expression. These non-coding RNAs can be divided into two main classes according to their length: short non-coding RNAs, such as microRNAs (miRNAs), and long non-coding RNAs (l...

It is well established that infection with Leishmania alters the host cell’s transcriptome. Since mammalian cells have multiple mechanisms to control gene expression, different molecules, such as noncoding RNAs, can be involved in this process. MicroRNAs have been extensively studied upon Leishmania infection, but whether long noncoding RNAs (lncRN...

Leishmaniases are neglected diseases that cause a large spectrum of clinical manifestations, from cutaneous to visceral lesions. The initial steps of the inflammatory response involve the phagocytosis of Leishmania and the parasite replication inside the macrophage phagolysosome. Melatonin, the darkness-signaling hormone, is involved in modulation...

Leishmania is a protozoan parasite that alternates its life cycle between the sand fly and the mammalian host macrophages, involving several environmental changes. The parasite responds to these changes by promoting a rapid metabolic adaptation through cellular signaling modifications that lead to transcriptional and post-transcriptional gene expre...

Background: Arginase is an enzyme that converts L-arginine to urea and L-ornithine, an essential substrate for the polyamine pathway supporting Leishmania (Leishmania) amazonensis replication and its survival in the mammalian host. L-arginine is also the substrate of macrophage nitric oxide synthase 2 (NOS2) to produce nitric oxide (NO) that kills...

Leishmania amazonensis is a protozoan that primarily causes cutaneous leishmaniasis in humans. The parasite relies on the amino acid arginine to survive within macrophages and establish infection, since it is a precursor for producing polyamines. On the other hand, arginine can be metabolized via nitric oxide synthase 2 (NOS2) to produce the microb...

Leishmania infectivity in melatonin-treated macrophages. BALB/c macrophages (2 × 105 cells) were pre-incubated for 1, 2, or 4 h with medium (untreated, white bar), vehicle (ethanol 0.0005%, gray bar), or 3 (blue bar) or 30 (light blue bar) nM of melatonin. After, macrophages were infected with L. amazonensis (MOI 5:1) and analyzed after 4 and 24 h....

Predicted mRNA-targets for miR-294. The miRNA-mRNA interaction was predicted using miRecord tools (http://c1.accurascience.com/miRecords/) based in the integration of various prediction tools: DIANA-microT, MicroInspector, miRanda, MirTarget2, miTarget, NBmiRTar, PicTar, PITA, RNA22, RNAhybrid, and TargetScan/TargertScanS.

Predicted mRNA-targets for miR-30e. The miRNA-mRNA interaction was predicted using miRecord tools (http://c1.accurascience.com/miRecords/) based in the integration of various prediction tools: DIANA-microT, MicroInspector, miRanda, MirTarget2, miTarget, NBmiRTar, PicTar, PITA, RNA22, RNAhybrid, and TargetScan/TargertScanS.

Predicted mRNA-targets for miR-302d. The miRNA-mRNA interaction was predicted using miRecord tools (http://c1.accurascience.com/miRecords/) based in the integration of various prediction tools: DIANA-microT, MicroInspector, miRanda, MirTarget2, miTarget, NBmiRTar, PicTar, PITA, RNA22, RNAhybrid, and TargetScan/TargertScanS.

Predicted mRNA-targets for miR-495. The miRNA-mRNA interaction was predicted using miRecord tools (http://c1.accurascience.com/miRecords/) based in the integration of various prediction tools: DIANA-microT, MicroInspector, miRanda, MirTarget2, miTarget, NBmiRTar, PicTar, PITA, RNA22, RNAhybrid, and TargetScan/TargertScanS.

Predicted mRNA-targets for miR-721. The miRNA-mRNA interaction was predicted using miRecord tools (http://c1.accurascience.com/miRecords/) based in the integration of various prediction tools: DIANA-microT, MicroInspector, miRanda, MirTarget2, miTarget, NBmiRTar, PicTar, PITA, RNA22, RNAhybrid, and TargetScan/TargertScanS.

miRNAs Up-Down Regulation in melatonin and infected L. amazonensis infected macrophages compared to uninfected. The relative up- and down-regulation of miRNAs, expressed as boundaries of 1.5 and −1.5 of Fold Regulation, respectively. p < 0.05 was considered statistically significant. p-value was determined based on two-tailed Student's t-test. The...

Predicted mRNA-targets for miR-181c. The miRNA-mRNA interaction was predicted using miRecord tools (http://c1.accurascience.com/miRecords/) based in the integration of various prediction tools: DIANA-microT, MicroInspector, miRanda, MirTarget2, miTarget, NBmiRTar, PicTar, PITA, RNA22, RNAhybrid, and TargetScan/TargertScanS.

Predicted mRNA-targets for miR-694. The miRNA-mRNA interaction was predicted using miRecord tools (http://c1.accurascience.com/miRecords/) based in the integration of various prediction tools: DIANA-microT, MicroInspector, miRanda, MirTarget2, miTarget, NBmiRTar, PicTar, PITA, RNA22, RNAhybrid, and TargetScan/TargertScanS.

Considering the limitations of the current leishmaniases chemotherapy and the lack of effective vaccines, the identification of novel drugs and/or vaccine approaches for the leishmaniases treatment and control is urgently required. In fact, a rational strategy for the parasite control can be based on the identification of essential metabolic pathwa...

Experimental validation of amastigotes differentiation. (A) The amastins LmxM.08.00760 (A), LmxM.0800800 (B) and LmxM.33.0960 (C) mRNA levels were based on quantification of the target and were normalized by gapdh expression in promastigotes (pro) and amastigotes (ama) of La-WT and La-arg-. The values are the mean ± SEM of three independent biologi...

Alignment of the nitric oxide synthase-like (LmxM.19.1450) from L. mexicana and their orthologs. Multiple alignment of the amino acid sequence of the nitric oxide synthase-like NOS-like of. L. amazonensis (Lam), L. mexicana (Lmx), L. donovani (Ldo), L. panamensis (Lpa), L. braziliensis (Lbr), L. major (Lmj) and L. infantum. The identical amino acid...

Experimental procedures for DAF-FM analysis in promastigotes by flow cytometry. The DAF-FM labeling of La-WT, La-arg- and La-arg-/+ARG promastigotes and analysis were performed as follows: (1) DAF-FM labeling followed by PI (propidium iodide) staining; (2) The data were acquired using a FACSCalibur (BD) flow cytometer followed by analysis of the re...

Experimental procedures for DAF-FM analysis from amastigotes by flow cytometry. DAF-FM labeling of La-WT, La-arg- and La-arg-/+ARG amastigotes and analysis were performed as follows: (1) DAF-FM labelling followed by PI (propidium iodide) staining; (2) The data were acquired on a FACSCalibur (BD) flow cytometer with analysis of the resulting pattern...

The establishment of Leishmania infection in macrophages can be modulated by non-coding microRNAs (miRNAs) by post-transcriptional regulation of genes through the complementary binding to the 3’ UTR of target mRNA. The production of Nitric Oxide (NO) by nitric oxide synthase 2 (NOS2) versus polyamines production via arginase 1 (ARG1) and ornithine...

Background: Leishmania uses the amino acid L-arginine as a substrate for arginase, enzyme that produces urea and ornithine, last precursor of polyamine pathway. This pathway is used by the parasite to replicate and it is essential to establish the infection in the mammalian host. L-arginine is not synthesized by the parasite, so its uptake occurs...

AAP3 protein growth curve of L. amazonensis promastigotes based on flow cytometry. The quantification of fluorescence intensity of anti-AAP3 was performed by image flow cytometry of permeabilized (total protein–gray) or non-permeabilized (plasma membrane protein–light gray) parasites. The values are the mean ± SEM of 3 independent biological replic...

AAP3 protein expression in La-WT, determined by Western blotting. Total extracts during the time-course of La-WT promastigotes and axenic amastigotes in the stationary phase, and La-WT promastigotes after starvation and supplementation with 400 μM L-arginine. Promastigotes were lysed and the proteins were separated by SDS-PAGE, transferred to nitro...