Journal: Archives of biochemistry and biophysics

Article Title: UDP-galactopyranose mutases from Leishmania species that cause visceral and cutaneous leishmaniasis

doi: 10.1016/j.abb.2013.08.014

Figure Lengend Snippet: Amino acid sequence analysis of T. cruzi UGM (TcUGM), L. infantum UGM (LinUGM), L. major UGM (LmUGM), L. mexicana UGM (LmxUGM), and A. fumigatus UGM (AfUGM). Identical amino acids are shown in red boxes and similar amino acids in white boxes. The amino acids involved in NADPH binding are marked with blue asterisks and those that interact with UDP-Galp are marked with orange carets. The conserved His in the histidine loop is marked with an arrow. The structural elements above the sequence are from the TcUGM structure (PDB code 4DSG). Clustal W was used to create the alignment and ESPript 2.2 to create the figure [44].

Article Snippet: The genes that code for L. mexicana and L. infantum UGMs were purchased from GeneScript (Piscataway, NJ).

Techniques: Sequencing, Binding Assay

Journal: Archives of biochemistry and biophysics

Article Title: UDP-galactopyranose mutases from Leishmania species that cause visceral and cutaneous leishmaniasis

doi: 10.1016/j.abb.2013.08.014

Figure Lengend Snippet: Steady-state kinetic parameters of eukaryotic UGMs.

Article Snippet: The genes that code for L. mexicana and L. infantum UGMs were purchased from GeneScript (Piscataway, NJ).

Techniques:

Journal: Archives of biochemistry and biophysics

Article Title: UDP-galactopyranose mutases from Leishmania species that cause visceral and cutaneous leishmaniasis

doi: 10.1016/j.abb.2013.08.014

Figure Lengend Snippet: Kinetic parameters for the reduction of eukaryotic UGMs by NADPH.

Article Snippet: The genes that code for L. mexicana and L. infantum UGMs were purchased from GeneScript (Piscataway, NJ).

Techniques:

Journal: Archives of biochemistry and biophysics

Article Title: UDP-galactopyranose mutases from Leishmania species that cause visceral and cutaneous leishmaniasis

doi: 10.1016/j.abb.2013.08.014

Figure Lengend Snippet: K D values for various ligands of L. infantum and L. mexicana UGMs.

Article Snippet: The genes that code for L. mexicana and L. infantum UGMs were purchased from GeneScript (Piscataway, NJ).

Techniques:

Journal: Clinical Epidemiology

Article Title: Epidemiology of visceral leishmaniasis

doi: 10.2147/CLEP.S44267

Figure Lengend Snippet: Disease types and transmission cycles of visceral leishmaniasis worldwide

Article Snippet: L. infantum , VL, CL , Med Europe, North Africa, Southwest Asia, People’s Republic of China , Rural, peri-domestic , Domestic dog, wild canids, domestic cat , P. (La.) ariasi , perniciosus , P. (La.) species; P.(Ad.) species.

Techniques: Transmission Assay, Plasmid Preparation

Journal: Genome Biology

Article Title: Modulation of gene expression in drug resistant Leishmania is associated with gene amplification, gene deletion and chromosome aneuploidy

doi: 10.1186/gb-2008-9-7-r115

Figure Lengend Snippet: Validation of DNA microarray expression data by qRT-PCR. The mean log10 ratios of selected genes from microarray expression data (grey bars) are compared to qRT-PCR data (black bars) for (a) L. infantum MTX20.5 and (b) L. major MTX60.4. The microarray data are the average of four biological replicates (with two dye swaps), while the qRT-PCR data are the average of three biological replicates repeated two times each. The asterisk indicates that the related gene transcript was not detected by qRT-PCR. The upper panel shows the expression of orthologous genes where the expression changes in the two species; the middle panel shows the modulation in the expression of FBT genes; the lower panel shows the expression of individual genes specific for each mutant.

Article Snippet: Completion of the L. major genome has allowed the generation of arrays containing 60-mer oligonucleotide probes designed by NimbleGen Systems [ , ] and in this work, we present the generation of a full genome DNA microarray composed of 70-mer oligonucleotide probes suitable for both L. major and L. infantum analysis (see Materials and methods for a full description of the arrays).

Techniques: Microarray, Expressing, Quantitative RT-PCR, Mutagenesis

Journal: Genome Biology

Article Title: Modulation of gene expression in drug resistant Leishmania is associated with gene amplification, gene deletion and chromosome aneuploidy

doi: 10.1186/gb-2008-9-7-r115

Figure Lengend Snippet: Extrachromosomal circular amplification of a genomic region of Leishmania chromosome 6 that includes the DHFR-TS locus. (a) Genomic organization of the DHFR-TS locus in both L. infantum MTX20.5 and L. major MTX60.4. Relative gene expression data (RNA) were determined using DNA microarrays and relative hybridization data were obtained by comparative genomic hybridization (DNA). Asterisks indicate that the microarray data of these genes were not found to be reliable. Direct repeats are shown with thick arrows and the approximate position of primers 6a and 6b are indicated with half arrows. (b) Chromosome size blot of Leishmania cells hybridized to a DHFR-TS probe. Sizes were determined using a yeast molecular weight marker (Biorad. Hercules, CA, USA). (c) Model for the formation of the extrachromosomal DHFR-TS circular DNA generated through homologous recombination between direct repeats (Figure S1 in Additional data file 2). (d) PCR with primers 6a and 6b to support the model shown in (c). Lane 1, L. infantum wild-type cells; lane 2, L. infantum MTX 20.5; lane 3, L. major wild-type cells; lane 4, L. major MTX60.4.

Article Snippet: Completion of the L. major genome has allowed the generation of arrays containing 60-mer oligonucleotide probes designed by NimbleGen Systems [ , ] and in this work, we present the generation of a full genome DNA microarray composed of 70-mer oligonucleotide probes suitable for both L. major and L. infantum analysis (see Materials and methods for a full description of the arrays).

Techniques: Amplification, Expressing, Hybridization, Microarray, Molecular Weight, Marker, Generated, Homologous Recombination

Journal: Vaccines

Article Title: Synthetic Peptides Elicit Strong Cellular Immunity in Visceral Leishmaniasis Natural Reservoir and Contribute to Long-Lasting Polyfunctional T-Cells in BALB/c Mice

doi: 10.3390/vaccines7040162

Figure Lengend Snippet: In vitro proliferation and IFN-γ production by T-lymphocytes in peripheral blood mononuclear cells (PBMC) of L. infantum naturally infected dogs after stimuli with predicted peptides. The spots represent indexes of T-cell responses to Leishmania infantum peptide pools in PBMC of naturally infected and asymptomatic dogs ( n = 5 in gray circles). Proliferation index of CD4 + ( a ) and CD8 + ( b ) lymphocytes after peptide pool stimuli. ( c ) Index of IFN-γ producing by CD4 + and CD8 + ( d ) lymphocytes after peptide pool stimuli. Indexes were calculated based on SLA (soluble Leishmania antigen)-stimulated (SC) cultures divided by the control culture (CC). Cut-off (dashed line) for responses was calculated by the control culture indexes (cut-off = 1).

Article Snippet: Thirty-eight L. infantum synthetic peptides (9–15 mer) were synthesized (Genscript, Piscataway, NJ, USA).

Techniques: In Vitro, Infection, Control

Journal: Vaccines

Article Title: Synthetic Peptides Elicit Strong Cellular Immunity in Visceral Leishmaniasis Natural Reservoir and Contribute to Long-Lasting Polyfunctional T-Cells in BALB/c Mice

doi: 10.3390/vaccines7040162

Figure Lengend Snippet: Spleen response after mice immunization with a cocktail of peptide-based vaccine and challenged with L. infantum . Cockt-1 promoted significant T-cell proliferation in spleens of immunized and challenged mice. ( a ) BALB/c mice were immunized three times and challenged with L. infantum promastigotes, four weeks after, splenocytes were obtained and T-cell proliferation and Th-1 cytokines were assessed through flow cytometry. ( b ) Representative plot of gating strategy to evaluate proliferation of T-cell subpopulations and IFN-γ and TNF-α–producing T-cells. ( c ) Plots represent index of T-cells proliferation (SC/CC ratio) of T-CD4 + and T-CD8 + cells after in vitro stimulation with SLA (soluble Leishmania antigen). ( d ) Plots represent the IFN-γ production indexes (SC/CC ratio) of T-CD4 + and T-CD8 + cells. ( e ) Plots represent the TNF-α production indexes (SC/CC ratio) of T-CD4 + and T-CD8 + cells after in vitro stimulation with SLA. Indexes were calculated based on SLA-stimulated (SC) cultures divided by the control culture (CC). Data are expressed as means ± SD of two independent experiments ( n = 8). The p -values represent the difference between the groups: * p- value < 0.05, ** p- value < 0.005, *** p- value < 0.0005.

Article Snippet: Thirty-eight L. infantum synthetic peptides (9–15 mer) were synthesized (Genscript, Piscataway, NJ, USA).

Techniques: Flow Cytometry, In Vitro, Control

Journal: Vaccines

Article Title: Synthetic Peptides Elicit Strong Cellular Immunity in Visceral Leishmaniasis Natural Reservoir and Contribute to Long-Lasting Polyfunctional T-Cells in BALB/c Mice

doi: 10.3390/vaccines7040162

Figure Lengend Snippet: Parasite burden and memory correlation in spleens of immunized mice with a cocktail of peptide-based vaccine challenged with Leishmania infantum . BALB/c mice were immunized three times and challenged with L. infantum promastigotes, and then four weeks after, they were sacrificed for further analyses. ( a ) Parasite burdens were assessed in the spleen through quantitative PCR and results are expressed in parasite DNA copies per spleen. ( b ) Representative plot of gating strategy to evaluate central memory (CM) and effector memory (EM) T-cells. CM cells were characterized by CD3 + CD4 + or CD8 + CD127 high CD44 high CD45RA low CD62L high C197 high markers, and EM cells were characterized by CD3 + CD4 + or CD8 + CD62L low CD44 high CD127 high C197 low after in vitro stimulation with SLA (soluble Leishmania antigen). Plots represent the frequency of CM T-cells ( c ) and EM T-cells ( d ) in mice splenocytes after in vitro stimulation with SLA. ( e ) Correlation analyses between parasite load and frequency of T CD4 + CM cells and T CD8 + EM cells are showed by Pearson’s r as means ± SD of two independent experiments ( n = 8), and p -values represent the difference between the groups: * p- value < 0.05, ** p- value < 0.005, *** p- value < 0.0005.

Article Snippet: Thirty-eight L. infantum synthetic peptides (9–15 mer) were synthesized (Genscript, Piscataway, NJ, USA).

Techniques: Real-time Polymerase Chain Reaction, In Vitro