fluoro max dyed red aqueous fluorescent particles  (Thermo Fisher)


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    Thermo Fisher fluoro max dyed red aqueous fluorescent particles
    Fluoro Max Dyed Red Aqueous Fluorescent Particles, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 97/100, based on 40 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 97 stars, based on 40 article reviews
    Price from $9.99 to $1999.99
    fluoro max dyed red aqueous fluorescent particles - by Bioz Stars, 2022-09
    97/100 stars

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    Thermo Fisher hygromycin b
    MMR and EGFP expression of transfected HCT116 cells. ( A ) Western blot of MMR proteins in HCT116 cells, showing a lack of MLH1 and MSH3 expression. ( B–D ) Fluorescent (GFP) and differential interference contrast (DIC) microscopic images (20×) of identical fields of variations of the (AAAG) 18 construct. ( B ) Images of positive control (MR-18-in-frame), negative controls (MR-18-[-1] out-of-frame and MR-18-[+1] out-of-frame) and experimental tetranucleotide constructs (AG-18-[-1] and AG-18-[+1]) cells 48 hours after transfection. Note EGFP expression was observed only in positive controls. ( C ) Positive control transfected cells showed 96%–97% EGFP expression after <t>hygromycin</t> B selection, whereas negative controls showed no visible fluorescence. ( D ) Cells transfected with the tetranucleotide construct in-frame with EGFP showed expression 48 hours after transfection. OF, out-of-frame; In, in-frame. AG, AAAG.
    Hygromycin B, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    97
    Thermo Fisher lps r60
    Titration of DS96 ( A ) and DS347 ( B ) to <t>LPS</t> <t>R60</t> and enthalpy change versus [LPA]:[LPS] molar ratio with isothermal titration calorimetry (ITC). The LPA solution (1 mM) was titrated in 3 µl portions to the LPS dispersion (0.05 mM), and the resulting enthalpy change was recorded.
    Lps R60, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    MMR and EGFP expression of transfected HCT116 cells. ( A ) Western blot of MMR proteins in HCT116 cells, showing a lack of MLH1 and MSH3 expression. ( B–D ) Fluorescent (GFP) and differential interference contrast (DIC) microscopic images (20×) of identical fields of variations of the (AAAG) 18 construct. ( B ) Images of positive control (MR-18-in-frame), negative controls (MR-18-[-1] out-of-frame and MR-18-[+1] out-of-frame) and experimental tetranucleotide constructs (AG-18-[-1] and AG-18-[+1]) cells 48 hours after transfection. Note EGFP expression was observed only in positive controls. ( C ) Positive control transfected cells showed 96%–97% EGFP expression after hygromycin B selection, whereas negative controls showed no visible fluorescence. ( D ) Cells transfected with the tetranucleotide construct in-frame with EGFP showed expression 48 hours after transfection. OF, out-of-frame; In, in-frame. AG, AAAG.

    Journal: Cellular and Molecular Gastroenterology and Hepatology

    Article Title: Tetranucleotide Microsatellite Mutational Behavior Assessed in Real Time: Implications for Future Microsatellite Panels

    doi: 10.1016/j.jcmgh.2020.01.006

    Figure Lengend Snippet: MMR and EGFP expression of transfected HCT116 cells. ( A ) Western blot of MMR proteins in HCT116 cells, showing a lack of MLH1 and MSH3 expression. ( B–D ) Fluorescent (GFP) and differential interference contrast (DIC) microscopic images (20×) of identical fields of variations of the (AAAG) 18 construct. ( B ) Images of positive control (MR-18-in-frame), negative controls (MR-18-[-1] out-of-frame and MR-18-[+1] out-of-frame) and experimental tetranucleotide constructs (AG-18-[-1] and AG-18-[+1]) cells 48 hours after transfection. Note EGFP expression was observed only in positive controls. ( C ) Positive control transfected cells showed 96%–97% EGFP expression after hygromycin B selection, whereas negative controls showed no visible fluorescence. ( D ) Cells transfected with the tetranucleotide construct in-frame with EGFP showed expression 48 hours after transfection. OF, out-of-frame; In, in-frame. AG, AAAG.

    Article Snippet: Selection by hygromycin B (50 mg/mL solution, cat. 10687-010; Life Technologies) started 1 day after transfection at the predetermined required dose for HCT116 cells (400 ug/mL) for 14 days.

    Techniques: Expressing, Transfection, Western Blot, Construct, Positive Control, Selection, Fluorescence

    Titration of DS96 ( A ) and DS347 ( B ) to LPS R60 and enthalpy change versus [LPA]:[LPS] molar ratio with isothermal titration calorimetry (ITC). The LPA solution (1 mM) was titrated in 3 µl portions to the LPS dispersion (0.05 mM), and the resulting enthalpy change was recorded.

    Journal: The Open Biochemistry Journal

    Article Title: Biophysical Mechanisms of the Neutralization of Endotoxins by Lipopolyamines

    doi: 10.2174/1874091X01307010082

    Figure Lengend Snippet: Titration of DS96 ( A ) and DS347 ( B ) to LPS R60 and enthalpy change versus [LPA]:[LPS] molar ratio with isothermal titration calorimetry (ITC). The LPA solution (1 mM) was titrated in 3 µl portions to the LPS dispersion (0.05 mM), and the resulting enthalpy change was recorded.

    Article Snippet: Briefly, phospholipid liposomes or LPS R60 were doubly labelled with the fluorescent phospholipid dyes 1,2-dipalmitoyl-sn -glycero-3-phosphoethanolamine-N -(7-nitro-2-1,3-benzoxadiazol-4-yl), ammonium salt (NBD-PE) and N -(lissamine rhodamine B sulfonyl)-phosphatidylethanolamine (Rh-PE) (Molecular Probes).

    Techniques: Titration, Isothermal Titration Calorimetry

    Gel-to-liquid crystalline phase transition of the hydrocarbon chains of LPS R60 at different concentrations of DS176 ( A ) and DS347 ( B ) by differential scanning calorimetry. The heat capacity C p is plotted versus temperature showing the enthalpy change at the phase transition temperature.

    Journal: The Open Biochemistry Journal

    Article Title: Biophysical Mechanisms of the Neutralization of Endotoxins by Lipopolyamines

    doi: 10.2174/1874091X01307010082

    Figure Lengend Snippet: Gel-to-liquid crystalline phase transition of the hydrocarbon chains of LPS R60 at different concentrations of DS176 ( A ) and DS347 ( B ) by differential scanning calorimetry. The heat capacity C p is plotted versus temperature showing the enthalpy change at the phase transition temperature.

    Article Snippet: Briefly, phospholipid liposomes or LPS R60 were doubly labelled with the fluorescent phospholipid dyes 1,2-dipalmitoyl-sn -glycero-3-phosphoethanolamine-N -(7-nitro-2-1,3-benzoxadiazol-4-yl), ammonium salt (NBD-PE) and N -(lissamine rhodamine B sulfonyl)-phosphatidylethanolamine (Rh-PE) (Molecular Probes).

    Techniques: Sublimation

    Gel-to-liquid crystalline phase transition of the hydrocarbon chains of LPS R60 at different concentrations of DS176 ( A ) and DS347 ( B ) by Fourier-transform infrared spectroscopy. The peak position of the symmetric vibration of the methylene groups ν s (CH 2 ) is plotted versus temperature. In the gel phase, the wavenumber values lie at 2850 cm -1 , in the liquid crystalline phase they are shifted to 2852.5 to 2853.0 cm -1 .

    Journal: The Open Biochemistry Journal

    Article Title: Biophysical Mechanisms of the Neutralization of Endotoxins by Lipopolyamines

    doi: 10.2174/1874091X01307010082

    Figure Lengend Snippet: Gel-to-liquid crystalline phase transition of the hydrocarbon chains of LPS R60 at different concentrations of DS176 ( A ) and DS347 ( B ) by Fourier-transform infrared spectroscopy. The peak position of the symmetric vibration of the methylene groups ν s (CH 2 ) is plotted versus temperature. In the gel phase, the wavenumber values lie at 2850 cm -1 , in the liquid crystalline phase they are shifted to 2852.5 to 2853.0 cm -1 .

    Article Snippet: Briefly, phospholipid liposomes or LPS R60 were doubly labelled with the fluorescent phospholipid dyes 1,2-dipalmitoyl-sn -glycero-3-phosphoethanolamine-N -(7-nitro-2-1,3-benzoxadiazol-4-yl), ammonium salt (NBD-PE) and N -(lissamine rhodamine B sulfonyl)-phosphatidylethanolamine (Rh-PE) (Molecular Probes).

    Techniques: Sublimation, Spectroscopy

    Reactive oxygen species (ROS) measurement in L. amazonensis promastigotes after SIVP. R0 and R60 promastigotes were cultivated in M199 medium (10% BFS, 26°C). The level of ROS was assessed by CellROX Deep Red Reagent (Thermo Fisher Scientific). After treatment, R0 and R60 cultures at late log phase (4th day) were analyzed by flow cytometry (BD Fortessa). (A) Percentage of cells producing ROS from R0 and R60. (B1,B2) Gates used to determine the percentage of R0 and R60 ROS producing cells, respectively. The bars represent the average of tree assays minus or plus the standard deviation from two independent tests. (C) Cytotoxic effects of trivalent tartrate antimony (Sb III ) on L. amazonensis (R0, R10, R40, and R60) promastigotes at the 4th day of cultivation. Parasites were incubated in M199 medium at 2 × 10 6 cells/mL into 24-well plates (26°C), either in the absence or presence of several concentrations of Sb III (18.71–374.32μM) for 48 h. The effective concentration required to decrease growth by 50% (EC 50 ) was determined using a Z1 Coulter Counter (Beckman Coulter, Fullerton, CA, USA). EC 50 values were determined from at least three independent measurements performed in triplicate, using the linear interpolation method. Statistical analyses were performed using GraphPad Prism Software v6.0, San Diego, CA. The EC 50 values were 47.09, 35.05, 29.52, 33.34 (μM; R0–R60, respectively; R2-values were > 0.95 for all cell lines). * p ≤ 0.05 by Two-Way ANOVA.

    Journal: Frontiers in Cellular and Infection Microbiology

    Article Title: Mapping Alterations Induced by Long-Term Axenic Cultivation of Leishmania amazonensis Promastigotes With a Multiplatform Metabolomic Fingerprint Approach

    doi: 10.3389/fcimb.2019.00403

    Figure Lengend Snippet: Reactive oxygen species (ROS) measurement in L. amazonensis promastigotes after SIVP. R0 and R60 promastigotes were cultivated in M199 medium (10% BFS, 26°C). The level of ROS was assessed by CellROX Deep Red Reagent (Thermo Fisher Scientific). After treatment, R0 and R60 cultures at late log phase (4th day) were analyzed by flow cytometry (BD Fortessa). (A) Percentage of cells producing ROS from R0 and R60. (B1,B2) Gates used to determine the percentage of R0 and R60 ROS producing cells, respectively. The bars represent the average of tree assays minus or plus the standard deviation from two independent tests. (C) Cytotoxic effects of trivalent tartrate antimony (Sb III ) on L. amazonensis (R0, R10, R40, and R60) promastigotes at the 4th day of cultivation. Parasites were incubated in M199 medium at 2 × 10 6 cells/mL into 24-well plates (26°C), either in the absence or presence of several concentrations of Sb III (18.71–374.32μM) for 48 h. The effective concentration required to decrease growth by 50% (EC 50 ) was determined using a Z1 Coulter Counter (Beckman Coulter, Fullerton, CA, USA). EC 50 values were determined from at least three independent measurements performed in triplicate, using the linear interpolation method. Statistical analyses were performed using GraphPad Prism Software v6.0, San Diego, CA. The EC 50 values were 47.09, 35.05, 29.52, 33.34 (μM; R0–R60, respectively; R2-values were > 0.95 for all cell lines). * p ≤ 0.05 by Two-Way ANOVA.

    Article Snippet: Evaluation of Cellular Oxidative Stress During SIVP Process To evaluate the cellular oxidative stress, promastigotes from R0 and R60 were cultivated in M199 medium (Gibco) supplemented with 10% of fetal bovine serum (Gibco) and cultivated at 26°C.

    Techniques: Flow Cytometry, Cytometry, Standard Deviation, Incubation, Concentration Assay, Software

    qPCR analysis for expression of 13 genes related to the SIVP process. All measured loci were classified in three clusters, according to its expression. Data represent fold changes for R10, R40, R60, in relation to R0. (A) Cluster 1-N-acetylglucosamine 1 phosphate, Cyclin, Isocitrate dehydrogenase, Small myristoylated protein, Cytosolic tryparedoxin peroxidase, Malic enzyme, Cyclopropane fatty acyl phospholipid synthase, ABC transporter subfamily G member 4. (B) Cluster 2-2-hydroxy-3-oxopropionate reductase, Mitochondrial tryparedoxin peroxidase. (C) Cluster 3-Trypanothione reductase, ABC transporter subfamily C, ABC transporter subfamily G member 2. Samples were analyzed in duplicates, from three different experiments.

    Journal: Frontiers in Cellular and Infection Microbiology

    Article Title: Mapping Alterations Induced by Long-Term Axenic Cultivation of Leishmania amazonensis Promastigotes With a Multiplatform Metabolomic Fingerprint Approach

    doi: 10.3389/fcimb.2019.00403

    Figure Lengend Snippet: qPCR analysis for expression of 13 genes related to the SIVP process. All measured loci were classified in three clusters, according to its expression. Data represent fold changes for R10, R40, R60, in relation to R0. (A) Cluster 1-N-acetylglucosamine 1 phosphate, Cyclin, Isocitrate dehydrogenase, Small myristoylated protein, Cytosolic tryparedoxin peroxidase, Malic enzyme, Cyclopropane fatty acyl phospholipid synthase, ABC transporter subfamily G member 4. (B) Cluster 2-2-hydroxy-3-oxopropionate reductase, Mitochondrial tryparedoxin peroxidase. (C) Cluster 3-Trypanothione reductase, ABC transporter subfamily C, ABC transporter subfamily G member 2. Samples were analyzed in duplicates, from three different experiments.

    Article Snippet: Evaluation of Cellular Oxidative Stress During SIVP Process To evaluate the cellular oxidative stress, promastigotes from R0 and R60 were cultivated in M199 medium (Gibco) supplemented with 10% of fetal bovine serum (Gibco) and cultivated at 26°C.

    Techniques: Real-time Polymerase Chain Reaction, Expressing

    Biochemical map of potential metabolic alterations occurred during axenization of L. amazonensis . Metabolites are represented by colored rectangles. Metabolites identified via metabolomic analysis are highlighted with a red contouring. The colored rectangles localized above/under metabolites represent the relative difference of abundance by comparing R10, R40, and R60 to R0 (left to right, respectively). Arrows represent reactions with enzymes, co-enzymes, metabolites or derivatives, indicated by their respective names or initials. Period-dash-period arrows indicate the occurrence of multiple reactions. Small dashes indicate interconnection of metabolic data between different metabolic pathways. The colors of arrows identify their respective pathways: 1. Black for energetic pathways (glycolytic and gluconeogenesis); 2. Yellow-green for pentose shunt (starting in G6P); 3. Light blue for amino acids biosynthesis; 4. Purple for fatty acids biosynthesis (receiving NADPH from pentose shunt and from malic enzyme reaction); 5. Light brown for β-alanine degradation (starting with ureidopropionate and culminating in α-keto-glutarate); 6. Dark brown for arginase and bright yellow for trypanothione (both pathways are connected in the production of putrescine); 7. Red for SAM or AdoMet pathway (interconnected to the Kennedy pathway, producer of phosphatidiletanolamine, phosphatidilcholine and phosphatidilserine); 8. Dark blue for glycine, serine and threonine biosynthesis. G6P, Glucose 6 phosphate; F6P, Fructose 6 phosphate; Fructose 1,6 bp, Fructose 6 bi-phosphate; PFK, phosphofructokinase; TPI, Triose phosphate isomerase; G3P, Glyceraldehyde 3-phosphate; 3PG, 3-phosphoglycerate; PEP, Phosphoenolpyruvate; UMP, Uridine monophosphate; UDP-GlcNac, Uridine diphosphate N- acetylglucosamine; HMG-CoA-Synthase, Hydroxymethylglutaryl-CoA synthase; NADPH, Nicotinamide adenine dinucleotide phosphate; ACP, Acyl carrier protein; TrypSyn, Trypanothione synthase; TrypRed, Trypanothione reductase; Gly, Glycine; Glu, Glutamine; ATP, Adenosine triphosphate; GS, Glutamine synthetase; SAH, S-adenosylhomocysteine; SAM, S-adennosylmethionine; PSS, phosphatidylserine synthase; PSD, Phosphatidylserine decarboxylase; CMP, Cytidine monophosphate; CK, Choline kinase; Pcyt1, Pyruvate carboxylase t1; CPT, Carnitine palmitoyltransferase; PLA, Phospholipase A; CTP, Phosphocholine cytidylyltransferase; ADP, Adenosine diphosphate; DAG, Diacylglycerol; PAF, Platelet-activating fator; PPi, Cytosolic pyrophosphate; PCP, Pyrrolidone carboxyl peptidase.

    Journal: Frontiers in Cellular and Infection Microbiology

    Article Title: Mapping Alterations Induced by Long-Term Axenic Cultivation of Leishmania amazonensis Promastigotes With a Multiplatform Metabolomic Fingerprint Approach

    doi: 10.3389/fcimb.2019.00403

    Figure Lengend Snippet: Biochemical map of potential metabolic alterations occurred during axenization of L. amazonensis . Metabolites are represented by colored rectangles. Metabolites identified via metabolomic analysis are highlighted with a red contouring. The colored rectangles localized above/under metabolites represent the relative difference of abundance by comparing R10, R40, and R60 to R0 (left to right, respectively). Arrows represent reactions with enzymes, co-enzymes, metabolites or derivatives, indicated by their respective names or initials. Period-dash-period arrows indicate the occurrence of multiple reactions. Small dashes indicate interconnection of metabolic data between different metabolic pathways. The colors of arrows identify their respective pathways: 1. Black for energetic pathways (glycolytic and gluconeogenesis); 2. Yellow-green for pentose shunt (starting in G6P); 3. Light blue for amino acids biosynthesis; 4. Purple for fatty acids biosynthesis (receiving NADPH from pentose shunt and from malic enzyme reaction); 5. Light brown for β-alanine degradation (starting with ureidopropionate and culminating in α-keto-glutarate); 6. Dark brown for arginase and bright yellow for trypanothione (both pathways are connected in the production of putrescine); 7. Red for SAM or AdoMet pathway (interconnected to the Kennedy pathway, producer of phosphatidiletanolamine, phosphatidilcholine and phosphatidilserine); 8. Dark blue for glycine, serine and threonine biosynthesis. G6P, Glucose 6 phosphate; F6P, Fructose 6 phosphate; Fructose 1,6 bp, Fructose 6 bi-phosphate; PFK, phosphofructokinase; TPI, Triose phosphate isomerase; G3P, Glyceraldehyde 3-phosphate; 3PG, 3-phosphoglycerate; PEP, Phosphoenolpyruvate; UMP, Uridine monophosphate; UDP-GlcNac, Uridine diphosphate N- acetylglucosamine; HMG-CoA-Synthase, Hydroxymethylglutaryl-CoA synthase; NADPH, Nicotinamide adenine dinucleotide phosphate; ACP, Acyl carrier protein; TrypSyn, Trypanothione synthase; TrypRed, Trypanothione reductase; Gly, Glycine; Glu, Glutamine; ATP, Adenosine triphosphate; GS, Glutamine synthetase; SAH, S-adenosylhomocysteine; SAM, S-adennosylmethionine; PSS, phosphatidylserine synthase; PSD, Phosphatidylserine decarboxylase; CMP, Cytidine monophosphate; CK, Choline kinase; Pcyt1, Pyruvate carboxylase t1; CPT, Carnitine palmitoyltransferase; PLA, Phospholipase A; CTP, Phosphocholine cytidylyltransferase; ADP, Adenosine diphosphate; DAG, Diacylglycerol; PAF, Platelet-activating fator; PPi, Cytosolic pyrophosphate; PCP, Pyrrolidone carboxyl peptidase.

    Article Snippet: Evaluation of Cellular Oxidative Stress During SIVP Process To evaluate the cellular oxidative stress, promastigotes from R0 and R60 were cultivated in M199 medium (Gibco) supplemented with 10% of fetal bovine serum (Gibco) and cultivated at 26°C.

    Techniques: Cycling Probe Technology, Proximity Ligation Assay

    Evaluation of metacyclogenesis in R0 and R60 cell lines. Promastigotes from R0 and R60 cell lines were cultivated in M199 medium (10% BFS, 26°C). R0 and R60 cultures at late log (4th day of growth) and stationary phases (7th day of growth) were analyzed by flow cytometry using BD Fortessa. (A) Percentages of promastigote metacyclic forms in R0 and R60 cell lines in late log (4 days) and stationary (7 days) phases. (B) B1 and B2 represent, respectively, R0 and R60 at the 4th day of growth. B3 and B4 represent, respectively, R0 and R60 at the 7th day of growth. The bars represent the average of two assays, minus and plus the standard deviation, in two independent analyses. Significant differences were investigated by Two-Way ANOVA.

    Journal: Frontiers in Cellular and Infection Microbiology

    Article Title: Mapping Alterations Induced by Long-Term Axenic Cultivation of Leishmania amazonensis Promastigotes With a Multiplatform Metabolomic Fingerprint Approach

    doi: 10.3389/fcimb.2019.00403

    Figure Lengend Snippet: Evaluation of metacyclogenesis in R0 and R60 cell lines. Promastigotes from R0 and R60 cell lines were cultivated in M199 medium (10% BFS, 26°C). R0 and R60 cultures at late log (4th day of growth) and stationary phases (7th day of growth) were analyzed by flow cytometry using BD Fortessa. (A) Percentages of promastigote metacyclic forms in R0 and R60 cell lines in late log (4 days) and stationary (7 days) phases. (B) B1 and B2 represent, respectively, R0 and R60 at the 4th day of growth. B3 and B4 represent, respectively, R0 and R60 at the 7th day of growth. The bars represent the average of two assays, minus and plus the standard deviation, in two independent analyses. Significant differences were investigated by Two-Way ANOVA.

    Article Snippet: Evaluation of Cellular Oxidative Stress During SIVP Process To evaluate the cellular oxidative stress, promastigotes from R0 and R60 were cultivated in M199 medium (Gibco) supplemented with 10% of fetal bovine serum (Gibco) and cultivated at 26°C.

    Techniques: Flow Cytometry, Cytometry, Standard Deviation

    Comparison of cytokine levels in spleen culture supernatants from BALB/c infected with R0 and R60 cell lines after 2 weeks of infection. For infection, 1 × 10 6 parasites were inoculated at the right footpad. Two weeks later, the animals were euthanized and RPMI medium (34°C, 5% C0 2 ) was used to cultivate spleen cells. The cultures were stimulated with soluble Leishmania antigen (SLA). Negative controls were non-stimulated spleen cells. 48 h after the stimulation, supernatant was collected and an ELISA assay was performed. (A) Levels of specific IFN-γ observed in spleen culture supernatants from BALB/c infected with R60 as compared to the levels produced by cells from mice infected with R0. (B) IL-10 levels in supernatant of splenocytes from R0 and R60. Bars represent mean values of three biological replicates from four infected BALB/c footpads, plus standard deviation of the mean. * indicates p ≤ 0.05 (Two-way ANOVA with Bonferroni post-test).

    Journal: Frontiers in Cellular and Infection Microbiology

    Article Title: Mapping Alterations Induced by Long-Term Axenic Cultivation of Leishmania amazonensis Promastigotes With a Multiplatform Metabolomic Fingerprint Approach

    doi: 10.3389/fcimb.2019.00403

    Figure Lengend Snippet: Comparison of cytokine levels in spleen culture supernatants from BALB/c infected with R0 and R60 cell lines after 2 weeks of infection. For infection, 1 × 10 6 parasites were inoculated at the right footpad. Two weeks later, the animals were euthanized and RPMI medium (34°C, 5% C0 2 ) was used to cultivate spleen cells. The cultures were stimulated with soluble Leishmania antigen (SLA). Negative controls were non-stimulated spleen cells. 48 h after the stimulation, supernatant was collected and an ELISA assay was performed. (A) Levels of specific IFN-γ observed in spleen culture supernatants from BALB/c infected with R60 as compared to the levels produced by cells from mice infected with R0. (B) IL-10 levels in supernatant of splenocytes from R0 and R60. Bars represent mean values of three biological replicates from four infected BALB/c footpads, plus standard deviation of the mean. * indicates p ≤ 0.05 (Two-way ANOVA with Bonferroni post-test).

    Article Snippet: Evaluation of Cellular Oxidative Stress During SIVP Process To evaluate the cellular oxidative stress, promastigotes from R0 and R60 were cultivated in M199 medium (Gibco) supplemented with 10% of fetal bovine serum (Gibco) and cultivated at 26°C.

    Techniques: Infection, Enzyme-linked Immunosorbent Assay, Produced, Mouse Assay, Standard Deviation

    aPs exposure by R0 and R60 promastigotes. (A) Profile of promastigotes grown for 4 days. (B) Profile of promastigotes grown for 7 days. 2 × 10 5 parasites from R0 and R60 were washed with PBS, suspended in binding buffer (BD-Anexin V-PE), and then incubated at room temperature for 15 min. At the moment of acquisition, 7AAD (PerCP-Cy5.5-A) was added as viability control, following manufacturer instructions. Data is shown as the difference between the geometric mean of the fluorescence intensity of unstained control samples ( Supplemental Figure 5 —Q1) compared to annexin V(PE) stained ones ( Supplemental Figure 5 —Q4). Viability was checked by 7AAD (PerCP-Cy5.5-A) parasites labeled ( Supplemental Figure 5 —Q2 and Q3). Data were collected in BD Fortessa and analyzed by FlowJo-X. At least 10,000 gated events were collected from each sample, in triplicates, from two essays. * p ≤ 0.05 Two-Way ANOVA.

    Journal: Frontiers in Cellular and Infection Microbiology

    Article Title: Mapping Alterations Induced by Long-Term Axenic Cultivation of Leishmania amazonensis Promastigotes With a Multiplatform Metabolomic Fingerprint Approach

    doi: 10.3389/fcimb.2019.00403

    Figure Lengend Snippet: aPs exposure by R0 and R60 promastigotes. (A) Profile of promastigotes grown for 4 days. (B) Profile of promastigotes grown for 7 days. 2 × 10 5 parasites from R0 and R60 were washed with PBS, suspended in binding buffer (BD-Anexin V-PE), and then incubated at room temperature for 15 min. At the moment of acquisition, 7AAD (PerCP-Cy5.5-A) was added as viability control, following manufacturer instructions. Data is shown as the difference between the geometric mean of the fluorescence intensity of unstained control samples ( Supplemental Figure 5 —Q1) compared to annexin V(PE) stained ones ( Supplemental Figure 5 —Q4). Viability was checked by 7AAD (PerCP-Cy5.5-A) parasites labeled ( Supplemental Figure 5 —Q2 and Q3). Data were collected in BD Fortessa and analyzed by FlowJo-X. At least 10,000 gated events were collected from each sample, in triplicates, from two essays. * p ≤ 0.05 Two-Way ANOVA.

    Article Snippet: Evaluation of Cellular Oxidative Stress During SIVP Process To evaluate the cellular oxidative stress, promastigotes from R0 and R60 were cultivated in M199 medium (Gibco) supplemented with 10% of fetal bovine serum (Gibco) and cultivated at 26°C.

    Techniques: Binding Assay, Incubation, Fluorescence, Staining, Labeling

    In vitro BMMΦ infection, lesion size development and tissue parasitism in footpads of BALB/c infected with R0 and R60 strains. (A) Percentage of infected bone marrow-derived macrophages. BALB/c bone marrow-derived macrophages (2 × 10 5 cells/ml) were infected with R0 or R60 grown for 7 days, at late stationary phase (5 parasites/macrophage). The cells were fixed and stained with Giemsa after 3 h of exposure (defined as 0 h time) and after 3, 12, 24, and 48 h post infection. The percentage (%) of infected macrophages was obtained by manual counting using an optical microscope. 300 cells at three different wells were evaluated from two different assays. (B) Evaluation of footpad lesion size from BALB/c mice infected with 1 × 10 6 L. amazonensis R0 or R60 stationary phase promastigotes. Lesion size was weekly measured with a caliper, and represent the size of infected paw minus the size of the non-infected one. Each point represents an average of lesion size from six infected mice plus the standard error (95% confidence interval). In detail, photographs of R0 and R60 infected paws at 8 weeks post-infection. (C) Limiting dilution assay, performed at 2nd week post-infection. Infected BALB/c footpads were homogenized in a glass homogenizer with M199 medium. The obtained pellet was resuspended in 500 μl of M199 medium and submitted to serial dilution (in duplicate) 1:10 in 96-well micro plates at final volume of 200 μL and incubated at 26°C for 15 days. The last dilution at which viable parasites was observed was considered the final titer. Results were expressed as negative logarithm of the parasites title. Bars represent mean values of three biological replicates from four infected BALB/c footpads plus standard deviation of the mean. * indicates p ≤ 0.05 (Two-way ANOVA with Bonferroni post-test).

    Journal: Frontiers in Cellular and Infection Microbiology

    Article Title: Mapping Alterations Induced by Long-Term Axenic Cultivation of Leishmania amazonensis Promastigotes With a Multiplatform Metabolomic Fingerprint Approach

    doi: 10.3389/fcimb.2019.00403

    Figure Lengend Snippet: In vitro BMMΦ infection, lesion size development and tissue parasitism in footpads of BALB/c infected with R0 and R60 strains. (A) Percentage of infected bone marrow-derived macrophages. BALB/c bone marrow-derived macrophages (2 × 10 5 cells/ml) were infected with R0 or R60 grown for 7 days, at late stationary phase (5 parasites/macrophage). The cells were fixed and stained with Giemsa after 3 h of exposure (defined as 0 h time) and after 3, 12, 24, and 48 h post infection. The percentage (%) of infected macrophages was obtained by manual counting using an optical microscope. 300 cells at three different wells were evaluated from two different assays. (B) Evaluation of footpad lesion size from BALB/c mice infected with 1 × 10 6 L. amazonensis R0 or R60 stationary phase promastigotes. Lesion size was weekly measured with a caliper, and represent the size of infected paw minus the size of the non-infected one. Each point represents an average of lesion size from six infected mice plus the standard error (95% confidence interval). In detail, photographs of R0 and R60 infected paws at 8 weeks post-infection. (C) Limiting dilution assay, performed at 2nd week post-infection. Infected BALB/c footpads were homogenized in a glass homogenizer with M199 medium. The obtained pellet was resuspended in 500 μl of M199 medium and submitted to serial dilution (in duplicate) 1:10 in 96-well micro plates at final volume of 200 μL and incubated at 26°C for 15 days. The last dilution at which viable parasites was observed was considered the final titer. Results were expressed as negative logarithm of the parasites title. Bars represent mean values of three biological replicates from four infected BALB/c footpads plus standard deviation of the mean. * indicates p ≤ 0.05 (Two-way ANOVA with Bonferroni post-test).

    Article Snippet: Evaluation of Cellular Oxidative Stress During SIVP Process To evaluate the cellular oxidative stress, promastigotes from R0 and R60 were cultivated in M199 medium (Gibco) supplemented with 10% of fetal bovine serum (Gibco) and cultivated at 26°C.

    Techniques: In Vitro, Infection, Derivative Assay, Staining, Microscopy, Mouse Assay, Limiting Dilution Assay, Serial Dilution, Incubation, Standard Deviation

    Relative expression of miRNAs and their target genes in fast and slow muscle. Groups C (control), F (fasting—10 days of fasting) and R60 (60 hours of refeeding). The data are expressed as fold change. Different letters indicate significant differences in expression between the groups (P

    Journal: PLoS ONE

    Article Title: Food restriction increase the expression of mTORC1 complex genes in the skeletal muscle of juvenile pacu (Piaractus mesopotamicus)

    doi: 10.1371/journal.pone.0177679

    Figure Lengend Snippet: Relative expression of miRNAs and their target genes in fast and slow muscle. Groups C (control), F (fasting—10 days of fasting) and R60 (60 hours of refeeding). The data are expressed as fold change. Different letters indicate significant differences in expression between the groups (P

    Article Snippet: Gene expression analysis of mRNAs and miRNAs involved in anabolic and catabolic processes Total RNA was extracted from fast and slow muscle samples in the C, F, R6 and R60 groups for mRNA analysis using TRIzol® Reagent (Life Technologies, USA), according to the manufacturer’s recommendations.

    Techniques: Expressing

    Relative mRNA expression of anabolic pathway and energetic metabolism components in slow muscle. Groups C (control), F (fasting—10 days of fasting), R6 (6 hours of refeeding) and R60 (60 hours of refeeding). The data are expressed as fold change. Different letters indicate significant differences in expression between the groups (P

    Journal: PLoS ONE

    Article Title: Food restriction increase the expression of mTORC1 complex genes in the skeletal muscle of juvenile pacu (Piaractus mesopotamicus)

    doi: 10.1371/journal.pone.0177679

    Figure Lengend Snippet: Relative mRNA expression of anabolic pathway and energetic metabolism components in slow muscle. Groups C (control), F (fasting—10 days of fasting), R6 (6 hours of refeeding) and R60 (60 hours of refeeding). The data are expressed as fold change. Different letters indicate significant differences in expression between the groups (P

    Article Snippet: Gene expression analysis of mRNAs and miRNAs involved in anabolic and catabolic processes Total RNA was extracted from fast and slow muscle samples in the C, F, R6 and R60 groups for mRNA analysis using TRIzol® Reagent (Life Technologies, USA), according to the manufacturer’s recommendations.

    Techniques: Expressing

    Relative mRNA expression of catabolic and anabolic pathway components in slow muscle. Groups C (control), F (fasting—10 days of fasting), R6 (6 hours of refeeding) and R60 (60 hours of refeeding). The data are expressed as fold change. Different letters indicate significant differences in expression between the groups (P

    Journal: PLoS ONE

    Article Title: Food restriction increase the expression of mTORC1 complex genes in the skeletal muscle of juvenile pacu (Piaractus mesopotamicus)

    doi: 10.1371/journal.pone.0177679

    Figure Lengend Snippet: Relative mRNA expression of catabolic and anabolic pathway components in slow muscle. Groups C (control), F (fasting—10 days of fasting), R6 (6 hours of refeeding) and R60 (60 hours of refeeding). The data are expressed as fold change. Different letters indicate significant differences in expression between the groups (P

    Article Snippet: Gene expression analysis of mRNAs and miRNAs involved in anabolic and catabolic processes Total RNA was extracted from fast and slow muscle samples in the C, F, R6 and R60 groups for mRNA analysis using TRIzol® Reagent (Life Technologies, USA), according to the manufacturer’s recommendations.

    Techniques: Expressing

    Relative mRNA expression of catabolic and anabolic pathway components in fast muscle. Groups C (control), F (fasting—10 days of fasting), R6 (6 hours of refeeding) and R60 (60 hours of refeeding). The data are expressed as fold change. Different letters indicate significant differences in expression between the groups (P

    Journal: PLoS ONE

    Article Title: Food restriction increase the expression of mTORC1 complex genes in the skeletal muscle of juvenile pacu (Piaractus mesopotamicus)

    doi: 10.1371/journal.pone.0177679

    Figure Lengend Snippet: Relative mRNA expression of catabolic and anabolic pathway components in fast muscle. Groups C (control), F (fasting—10 days of fasting), R6 (6 hours of refeeding) and R60 (60 hours of refeeding). The data are expressed as fold change. Different letters indicate significant differences in expression between the groups (P

    Article Snippet: Gene expression analysis of mRNAs and miRNAs involved in anabolic and catabolic processes Total RNA was extracted from fast and slow muscle samples in the C, F, R6 and R60 groups for mRNA analysis using TRIzol® Reagent (Life Technologies, USA), according to the manufacturer’s recommendations.

    Techniques: Expressing

    Relative mRNA expression of anabolic pathway and energetic metabolism components in fast muscle. Groups C (control), F (fasting—10 days of fasting), R6 (6 hours of refeeding) and R60 (60 hours of refeeding). The data are expressed as fold change. Different letters indicate significant differences in expression between the groups (P

    Journal: PLoS ONE

    Article Title: Food restriction increase the expression of mTORC1 complex genes in the skeletal muscle of juvenile pacu (Piaractus mesopotamicus)

    doi: 10.1371/journal.pone.0177679

    Figure Lengend Snippet: Relative mRNA expression of anabolic pathway and energetic metabolism components in fast muscle. Groups C (control), F (fasting—10 days of fasting), R6 (6 hours of refeeding) and R60 (60 hours of refeeding). The data are expressed as fold change. Different letters indicate significant differences in expression between the groups (P

    Article Snippet: Gene expression analysis of mRNAs and miRNAs involved in anabolic and catabolic processes Total RNA was extracted from fast and slow muscle samples in the C, F, R6 and R60 groups for mRNA analysis using TRIzol® Reagent (Life Technologies, USA), according to the manufacturer’s recommendations.

    Techniques: Expressing