ml 265  (MedChemExpress)

Journal: mSphere

Article Title: Endosymbiosis in trypanosomatids: the bacterium regulates the intermediate and oxidative metabolism of the host cell

doi: 10.1128/msphere.00457-25

Figure Lengend Snippet: Proliferation, viability, and glucose consumption of A. deanei . ( A ) Proliferation curve of wild-type (AdWt) and aposymbiotic (AdApo) strains. AdWt in Warren with FBS (control), blue line and bar (●), AdWt in SDM80 medium with glucose, brown line and bar (■), AdWt in SDM80 medium with proline, light blue line and bar (▲), AdWt in SDM80 medium – fasting, orange line and bar (▼), AdApo in control medium, blue patterned line and bar (○), AdApo in SDM80 medium with glucose, brown patterned line and bar (□), AdApo in SDM80 medium with proline, light blue patterned line and bar (Δ), AdApo in SDM80 medium, fasting, orange patterned line and bar (∇). ( B ) Area under the curve of proliferation. ( C ) Cell viability after cultivation for 24 and 60 h in different nutritional conditions. ( D ) Glucose consumption in Warren medium supplemented with 20 mM glucose. Means ± SEM and statistical analyses, area under the curve for ( A ), ( B ) one-way ANOVA with Holm-Šídák’s multiple comparisons test. ( D ) Unpaired t -test. P -value **** (<0.0001), ** (<0.005), * (<0.05). ( A–D ) N = 3.

Article Snippet: Wild-type (AdWt) (ATCC PRA-265) and aposymbiotic (AdApo) (ATCC 30969) strains of A. deanei were grown at 28°C in Warren’s complex medium (3.7% Sigma-Aldrich BHI broth, brain and heart infusion) , containing 0.1% folic acid, 25 μg/mL hemin, plus 10% supplementation with fetal bovine serum (FBS) (Vitrocell, Embriolife, Brazil) for 24 h, which corresponds to exponential cell growth.

Techniques: Control

Journal: mSphere

Article Title: Endosymbiosis in trypanosomatids: the bacterium regulates the intermediate and oxidative metabolism of the host cell

doi: 10.1128/msphere.00457-25

Figure Lengend Snippet: Morphological analysis of A. deanei cultivated in different culture media. ( A–D ) Wild-type strain (AdWt): ( A ) control medium, ( B ) SDM80 with glucose, ( C ) SDM80 with proline, ( D ) fasting. ( E–H ) Aposymbiotic strain (AdApo): ( E ) control medium, ( F ) SDM80 with glucose, ( G ) SDM80 with proline, ( H ) fasting. White arrows indicate rounded cells or protozoa with a reduced length of the cell body presenting a wrinkled surface. White arrowheads indicate flagellar shortening. ( I ) Morphometric data based on SEM analyses. ( J ) Parameters for morphometric analysis. Medians and quartiles were calculated, and statistical analyses were performed using a two-way ANOVA and a Holm-Šídák multiple comparisons test for comparisons between conditions and a one-way ANOVA for comparisons between strains. P -value **** (<0.0001), *** (<0.001), ** (<0.005), * (<0.05). N = 100 cells. Scale bars: 1 µm.

Article Snippet: Wild-type (AdWt) (ATCC PRA-265) and aposymbiotic (AdApo) (ATCC 30969) strains of A. deanei were grown at 28°C in Warren’s complex medium (3.7% Sigma-Aldrich BHI broth, brain and heart infusion) , containing 0.1% folic acid, 25 μg/mL hemin, plus 10% supplementation with fetal bovine serum (FBS) (Vitrocell, Embriolife, Brazil) for 24 h, which corresponds to exponential cell growth.

Techniques: Control

Journal: mSphere

Article Title: Endosymbiosis in trypanosomatids: the bacterium regulates the intermediate and oxidative metabolism of the host cell

doi: 10.1128/msphere.00457-25

Figure Lengend Snippet: High-resolution respirometry and intracellular ATP quantification of A. deanei . ( A ) Representative graph of O 2 flux per cell of wild-type (AdWt) and aposymbiotic (AdApo) strains. U indicates the addition of FCCP. ( B ) Basal respiration. ( C ) Respiration in the presence of glucose and proline. ( D ) Uncoupled respiration using FCCP. ( E ) Residual respiration (Rox). ( F ) Routine respiration. ( G ) Maximum capacity. ( H ) Reserve capacity. ( I ) ATP is produced at the substrate level after using KCN. ( J ) ATP produced by mitochondrial OxPhos. Means ± SEM and statistical analyses performed by Unpaired t -test ( B–I ), One-Way ANOVA ( C–H ), and two-way ANOVA with Holm-Šídák’s multiple comparisons test. P -value **** (<0.0001), *** (<0.001), ** (0.005), * (<0.05) ( J ).

Article Snippet: Wild-type (AdWt) (ATCC PRA-265) and aposymbiotic (AdApo) (ATCC 30969) strains of A. deanei were grown at 28°C in Warren’s complex medium (3.7% Sigma-Aldrich BHI broth, brain and heart infusion) , containing 0.1% folic acid, 25 μg/mL hemin, plus 10% supplementation with fetal bovine serum (FBS) (Vitrocell, Embriolife, Brazil) for 24 h, which corresponds to exponential cell growth.

Techniques: Produced

Journal: mSphere

Article Title: Endosymbiosis in trypanosomatids: the bacterium regulates the intermediate and oxidative metabolism of the host cell

doi: 10.1128/msphere.00457-25

Figure Lengend Snippet: Proton ( 1 H) 1D-NMR spectra and incorporation ratios of [ 13 C-U]-glucose in metabolic end-products excreted and proteomic analysis of A. deanei . ( A ) Representative spectra of AdApo and AdWt, respectively. ( B, C ) Quantification of metabolites. E, ethanol; S, succinate; and A, acetate. ( D ) Venn diagram of the proteomics of the total extract of A. deanei , the AdWt (1) and AdApo (2) strains using the PLP program. This diagram shows the number of proteins found in each proteome, from the wild strain (1,229, pink) and the aposymbiotic strain (1,086, blue), the proteins common to both groups (1,031, purple) and those exclusive to the wild strain (198) or the aposymbiotic strain (55). ( E ) Comparative proteomic analysis between AdWt and AdApo strains. Pink dots: proteins with P Value less than 0.05; purple dots: low-abundant proteins; blue dots: proteins more abundant in one strain than the other, with low statistical power; cyan dots: proteins equally abundant in both A. deanei strains. ( A–C ) Means ± SEM and statistical analyses performed by Unpaired t -test. P -value **** (<0.0001). N = 5 to AdWt and N = 6 to AdApo.

Article Snippet: Wild-type (AdWt) (ATCC PRA-265) and aposymbiotic (AdApo) (ATCC 30969) strains of A. deanei were grown at 28°C in Warren’s complex medium (3.7% Sigma-Aldrich BHI broth, brain and heart infusion) , containing 0.1% folic acid, 25 μg/mL hemin, plus 10% supplementation with fetal bovine serum (FBS) (Vitrocell, Embriolife, Brazil) for 24 h, which corresponds to exponential cell growth.

Techniques:

Journal: mSphere

Article Title: Endosymbiosis in trypanosomatids: the bacterium regulates the intermediate and oxidative metabolism of the host cell

doi: 10.1128/msphere.00457-25

Figure Lengend Snippet: Schematic representation illustrates glucose and L-proline metabolism in wild-type ( A ), aposymbiotic ( B ) A. deanei strains, and the procyclic form of T. brucei ( C ) adapted . This diagram provides a detailed overview of metabolic pathways and their enzymatic steps in these organisms. Pink arrows indicate glucose metabolism, whereas blue arrows represent L-proline metabolism. Excreted end products are displayed in the extracellular region: acetate for the wild-type strain (AdWt) and alcohol, succinate, and acetate for the aposymbiotic strain (AdApo). Reversible reactions are depicted only in their presumed or demonstrated direction, and dashed arrows indicate steps occurring at background levels. The schematic highlights key organelles, including the glycosome and mitochondria, along with the tricarboxylic acid (TCA) cycle. Abbreviations for metabolites include: G-6-P, glucose-6-phosphate; F-6-P, fructose-6-phosphate; F-B-P, fructose-1,6-bisphosphate; DHAP, dihydroxyacetone phosphate; G-3-P, glyceraldehyde-3-phosphate; 1,3BPGA, 1,3-bisphosphoglycerate; 3-PGA, 3-phosphoglycerate; 2-PGA, 2-phosphoglycerate; PEP, phosphoenolpyruvate; ACD, acetaldehyde; Cit, citrate; IsoCit, isocitrate; αKet, α-ketoglutarate; SucCoA, succinyl-CoA; Fum, fumarate; Mal, malate; Oxac, oxaloacetate; P5C, pyrroline-5-carboxylate; γSAG, glutamate-γ-semialdehyde; Q, ubiquinone pool; C, cytochrome c; and CoASH, coenzyme A. The enzymes involved are: 1, Hexokinase; 2, glucose-6-phosphate isomerase; 3, phosphofructokinase; 4, aldolase; 5, triosephosphate isomerase; 6, glyceraldehyde-3-phosphate dehydrogenase; 7, cytosolic phosphoglycerate kinase; 8, phosphoglycerate mutase; 9, enolase; 10, pyruvate kinase; 11, pyruvate decarboxylase; 12, alcohol dehydrogenase; 13, pyruvate dehydrogenase complex; 14, citrate synthase; 15, aconitase; 16, NADP-dependent isocitrate dehydrogenase; 17, α-ketoglutarate dehydrogenase complex; 18, succinyl-CoA synthetase; 19, succinate dehydrogenase (complex II); 20, mitochondrial fumarate hydratase; 21, mitochondrial malate dehydrogenase; 22, rotenone-insensitive NADH dehydrogenase; 23, Complex III; 24, proline dehydrogenase; 25, spontaneous reaction; 26, Δ−1-pyrroline-5-carboxylate dehydrogenase; 27, glutamate dehydrogenase; 28, acetate:succinate CoA-transferase (ASCT); 29, fumarate reductase; 30, alternative oxidase (AOX); 31, pyruvate dikinase; 32, phosphoenolpyruvate carboxylase; 33, acetyl-CoA synthetase; 34, glycerol-3-phosphate dehydrogenase; 35, glycerol kinase; 36, mitochondrial glycerol-3-phosphate dehydrogenase; 37, lactate dehydrogenase; 38, non-enzymatic reaction; 39, NADPH-dependent methylglyoxal reductase; and 40, L-alanine aminotransferase. Respiratory chain components include CIV, cytochrome c oxidase complex, and CV, F0/F1-ATP synthase.

Article Snippet: Wild-type (AdWt) (ATCC PRA-265) and aposymbiotic (AdApo) (ATCC 30969) strains of A. deanei were grown at 28°C in Warren’s complex medium (3.7% Sigma-Aldrich BHI broth, brain and heart infusion) , containing 0.1% folic acid, 25 μg/mL hemin, plus 10% supplementation with fetal bovine serum (FBS) (Vitrocell, Embriolife, Brazil) for 24 h, which corresponds to exponential cell growth.

Techniques: