Journal: F1000Research

Article Title: Plackett-Burman design in the biosynthesis of silver nanoparticles with Mutisia acuminatta (Chinchircoma) and preliminary evaluation of its antibacterial activity

doi: 10.12688/f1000research.140883.1

Figure Lengend Snippet: Representation of antibacterial effect against certified strains of Staphylococcus aureus ATCC 25923 and Escherichia coli ATCC 33876: (a) MetOH, (b) AgNPs - FM, (c) FM and (d) EMT (5%).

Article Snippet: shows the results obtained by the inhibition halos method for the sensitivity analysis of the samples evaluated against the bacterial strains Escherichia coli ATCC 33876 and Staphylococcus aureus ATCC 25923.

Techniques:

Journal: F1000Research

Article Title: Plackett-Burman design in the biosynthesis of silver nanoparticles with Mutisia acuminatta (Chinchircoma) and preliminary evaluation of its antibacterial activity

doi: 10.12688/f1000research.140883.1

Figure Lengend Snippet: Agar well assays for the evaluation of antibacterial activity against Escherichia coli ATCC 33876: (A) EMT (5%), (B) FM, (C) AgNPs-FM and (D) Methanol (blank).

Article Snippet: shows the results obtained by the inhibition halos method for the sensitivity analysis of the samples evaluated against the bacterial strains Escherichia coli ATCC 33876 and Staphylococcus aureus ATCC 25923.

Techniques: Activity Assay

Journal: Nanomaterials

Article Title: Biological Performances of Plasmonic Biohybrids Based on Phyto-Silver/Silver Chloride Nanoparticles

doi: 10.3390/nano11071811

Figure Lengend Snippet: Antibacterial activity expressed as a diameter of the inhibition growth zone (IGZ) against Staphylococcus aureus ATTC 2592 and Escherichia coli ATCC 8738.

Article Snippet: Antimicrobial activity of the samples was tested against pathogenic Gram (-) bacteria Escherichia coli ATCC 8738 and Gram (+) bacteria Staphylococcus aureus ATTC 25923.

Techniques: Activity Assay, Inhibition

Journal: Engineering in Life Sciences

Article Title: Analysis of intracellular metabolites of Corynebacterium glutamicum at high cell density with automated sampling and filtration and assessment of engineered enzymes for effective l ‐lysine production

doi: 10.1002/elsc.201600163

Figure Lengend Snippet: Filtrate volumes of samples and wash solution at different cell densities for metabolite sampling using automated fast filtration directly from bioreactor cultures. (A) C. glutamicum wild type. (B) C. glutamicum LP917. Mean values and standard deviations were calculated from three technical replicates.

Article Snippet: The bacterial strains cultivated in this study were the wild‐type strain C. glutamicum ATCC 13032 and the l ‐lysine‐producing mutant strain C. glutamicum LP917, which has the point mutation Q298G in the lysC gene and N917G in the PEPC gene as described previously 17 , 19 .

Techniques: Sampling, Filtration

Journal: Engineering in Life Sciences

Article Title: Analysis of intracellular metabolites of Corynebacterium glutamicum at high cell density with automated sampling and filtration and assessment of engineered enzymes for effective l ‐lysine production

doi: 10.1002/elsc.201600163

Figure Lengend Snippet: Cultivation profiles of C. glutamicum wild type and LP917 during batch cultures. (A) Biomass growth as optical density OD660. Means and standard deviations of optical density were calculated from three technical replicates. (B) Extracellular Lysine concentration. Mean values were calculated from two analytical replicates and the whiskers show the upper and lower value, respectively.

Article Snippet: The bacterial strains cultivated in this study were the wild‐type strain C. glutamicum ATCC 13032 and the l ‐lysine‐producing mutant strain C. glutamicum LP917, which has the point mutation Q298G in the lysC gene and N917G in the PEPC gene as described previously 17 , 19 .

Techniques: Concentration Assay

Journal: Engineering in Life Sciences

Article Title: Analysis of intracellular metabolites of Corynebacterium glutamicum at high cell density with automated sampling and filtration and assessment of engineered enzymes for effective l ‐lysine production

doi: 10.1002/elsc.201600163

Figure Lengend Snippet: Time points for intracellular metabolite sampling using automated fast filtration

Article Snippet: The bacterial strains cultivated in this study were the wild‐type strain C. glutamicum ATCC 13032 and the l ‐lysine‐producing mutant strain C. glutamicum LP917, which has the point mutation Q298G in the lysC gene and N917G in the PEPC gene as described previously 17 , 19 .

Techniques: Sampling, Filtration

Journal: Engineering in Life Sciences

Article Title: Analysis of intracellular metabolites of Corynebacterium glutamicum at high cell density with automated sampling and filtration and assessment of engineered enzymes for effective l ‐lysine production

doi: 10.1002/elsc.201600163

Figure Lengend Snippet: Concentration profiles of intracellular metabolites during batch cultures of C. glutamicum wild type and C. glutamicum LP917. The sampling points are displayed by the biomass concentrations according to Table ​Table1.1. Mean values and standard deviations were calculated from three technical replicate samples.

Article Snippet: The bacterial strains cultivated in this study were the wild‐type strain C. glutamicum ATCC 13032 and the l ‐lysine‐producing mutant strain C. glutamicum LP917, which has the point mutation Q298G in the lysC gene and N917G in the PEPC gene as described previously 17 , 19 .

Techniques: Concentration Assay, Sampling

Journal: Engineering in Life Sciences

Article Title: Analysis of intracellular metabolites of Corynebacterium glutamicum at high cell density with automated sampling and filtration and assessment of engineered enzymes for effective l ‐lysine production

doi: 10.1002/elsc.201600163

Figure Lengend Snippet: Concentration profiles of intracellular metabolites during batch cultures of C. glutamicum wild type and C. glutamicum LP917. The sampling points are displayed by the biomass concentrations according to Table ​Table1.1. Mean values and standard deviations were calculated from three technical replicate samples.

Article Snippet: The bacterial strains cultivated in this study were the wild‐type strain C. glutamicum ATCC 13032 and the l ‐lysine‐producing mutant strain C. glutamicum LP917, which has the point mutation Q298G in the lysC gene and N917G in the PEPC gene as described previously 17 , 19 .

Techniques: Concentration Assay, Sampling

Journal: Engineering in Life Sciences

Article Title: Analysis of intracellular metabolites of Corynebacterium glutamicum at high cell density with automated sampling and filtration and assessment of engineered enzymes for effective l ‐lysine production

doi: 10.1002/elsc.201600163

Figure Lengend Snippet: Comparison of intracellular amino acid levels of C. glutamicum wild type and LP917. The relative difference δ describes the normalized overall difference for all sampling points during the exponential growth phase: positive values represent higher intracellular levels in the lysine‐producing strain LP917 compared to the wild type. Error bars represent the normalized cumulated errors of the five sampling points during exponential growth.

Article Snippet: The bacterial strains cultivated in this study were the wild‐type strain C. glutamicum ATCC 13032 and the l ‐lysine‐producing mutant strain C. glutamicum LP917, which has the point mutation Q298G in the lysC gene and N917G in the PEPC gene as described previously 17 , 19 .

Techniques: Sampling

Journal: Engineering in Life Sciences

Article Title: Analysis of intracellular metabolites of Corynebacterium glutamicum at high cell density with automated sampling and filtration and assessment of engineered enzymes for effective l ‐lysine production

doi: 10.1002/elsc.201600163

Figure Lengend Snippet: In vitro inhibition profiles of aspartate kinase for lysine, threonine and concerted inhibition of both amino acids according to 17. The shaded areas represent the in vivo concentration ranges of intracellular lysine   and threonine   determined in this study (25% to 75% quartile of the concentrations determined during exponential growth). (A) C. glutamicum wild type. (B) C. glutamicum LP917.

Article Snippet: The bacterial strains cultivated in this study were the wild‐type strain C. glutamicum ATCC 13032 and the l ‐lysine‐producing mutant strain C. glutamicum LP917, which has the point mutation Q298G in the lysC gene and N917G in the PEPC gene as described previously 17 , 19 .

Techniques: In Vitro, Inhibition, In Vivo, Concentration Assay

Journal: Engineering in Life Sciences

Article Title: Analysis of intracellular metabolites of Corynebacterium glutamicum at high cell density with automated sampling and filtration and assessment of engineered enzymes for effective l ‐lysine production

doi: 10.1002/elsc.201600163

Figure Lengend Snippet: In vitro inhibition profiles of phosphoenolpyruvate carboxylase for Aspartate and malate, according to 19. The shaded areas represent the in vivo concentration ranges of intracellular Aspartate   and malate   determined in this study (25% to 75% quartile of the concentrations determined during exponential growth). (A) C. glutamicum wild type. (B) C. glutamicum LP917.

Article Snippet: The bacterial strains cultivated in this study were the wild‐type strain C. glutamicum ATCC 13032 and the l ‐lysine‐producing mutant strain C. glutamicum LP917, which has the point mutation Q298G in the lysC gene and N917G in the PEPC gene as described previously 17 , 19 .

Techniques: In Vitro, Inhibition, In Vivo, Concentration Assay

Journal: Cells

Article Title: Flow Cytometric Detection of Waterborne Bacteria Metabolic Response to Anthropogenic Chemical Inputs to Aquatic Ecosystems

doi: 10.3390/cells14050352

Figure Lengend Snippet: Cytograms from Escherichia coli ( E. coli ) staining controls for metabolic activity of live (left column) or heat-killed (right column) cells. Unstained bacteria and auto-fluorescent bacteria, concentrated near the origin, were excluded (out-gated; round gates above) for statistical computations. Bacteria stained with CDFA (5(6)-carboxyfluorescein diacetate) yield a fluorescent product because of hydrolysis by esterases (FL1-H axis; green fluorescence) shown here as populations focused at the bottom portion of the cytograms. Bacteria stained with propidium iodide (that fluoresces when intercalated into double-stranded nucleic acids of membrane-damaged cells) (FL3-H axis; red fluorescence) shown here as populations focused on the left portions of the cytograms.

Article Snippet: For use as a staining control for known dead cells, with each water sample, a 10 μL aliquot of an overnight Escherichia coli (ATCC 35218) at 2 × 10 6 cells mL −1 was heat-killed in a water bath at 75 °C for at least 30 min.

Techniques: Staining, Activity Assay, Bacteria, Fluorescence, Membrane

Journal: Cells

Article Title: Flow Cytometric Detection of Waterborne Bacteria Metabolic Response to Anthropogenic Chemical Inputs to Aquatic Ecosystems

doi: 10.3390/cells14050352

Figure Lengend Snippet: Representative flow cytometric zebra plots display representative analyses of metabolic activity in strains of bacteria that were laboratory-exposed to organic wastewater compounds. Percentages of stained cells are presented, whereby the upper gate is that of the metabolically inactive cells and the right-hand gate shows the metabolically active percentage. In panels ( A , B ): Escherichia coli was incubated with atrazine and tylosin, respectively. ( C , D ): Streptococcus suis was incubated with trenbolone and butylparaben, respectively. ( E , F ): Streptococcus dysgalactiae was incubated with atrazine and tylosin, respectively. Staining with 5(6)-carboxyfluorescein diacetate yields a fluorescent product upon hydrolysis by esterases (horizontal axis; FL1-H; green fluorescence), and propidium iodide counter-stains nucleic acids in membrane-damaged cells (vertical axis; FL3-H; red fluorescence). Unstained or auto-fluorescent particles were out-gated for the analyses (e.g., ungated population near the origin) .

Article Snippet: For use as a staining control for known dead cells, with each water sample, a 10 μL aliquot of an overnight Escherichia coli (ATCC 35218) at 2 × 10 6 cells mL −1 was heat-killed in a water bath at 75 °C for at least 30 min.

Techniques: Activity Assay, Bacteria, Staining, Metabolic Labelling, Incubation, Fluorescence, Membrane

Journal: Cells

Article Title: Flow Cytometric Detection of Waterborne Bacteria Metabolic Response to Anthropogenic Chemical Inputs to Aquatic Ecosystems

doi: 10.3390/cells14050352

Figure Lengend Snippet: Statistical main effects and two-way interactions of inhibition of reductase activity measured by flow cytometry after exponential phase bacterial strains were exposed to organic wastewater compounds (OWCs) 1 that had been, or were likely to occur, in Buffalo National River 2 .

Article Snippet: For use as a staining control for known dead cells, with each water sample, a 10 μL aliquot of an overnight Escherichia coli (ATCC 35218) at 2 × 10 6 cells mL −1 was heat-killed in a water bath at 75 °C for at least 30 min.

Techniques: Inhibition, Activity Assay, Flow Cytometry, Bacteria, Concentration Assay, Incubation

Journal: Cell

Article Title: Vaginal Lactobacillus fatty acid response mechanisms reveal a metabolite-targeted strategy for bacterial vaginosis treatment

doi: 10.1016/j.cell.2024.07.029

Figure Lengend Snippet: FGT Lactobacillus OhyA enzymes are functional and physiologically active (A) OhyA protein phylogenetic tree for representative orthologs from the indicated species (see ). Starred leaf tips indicate FGT Lactobacillus orthologs; ∗ indicates confirmed OA-induced ortholog ( Figure 2 A). (B) Diagram of OhyA9 enzymatic activity with OA substrate and 10-hydroxystearic acid (10-HSA or h 18:0) product. (C) Extracted ion chromatograms from supernatants of ohyA -gene deleted S. aureus complemented with empty vector ( ΔSaohyA /empty vector), SaohyA -expressing plasmid ( ΔSaohyA /p SaohyA ), LCRIS_00558-expressing plasmid ( ΔSaohyA /pLCRIS_00558), or LCRIS_00661-expressing plasmid ( ΔSaohyA /pLCRIS_00661), cultured with OA for 1 h. Annotated peaks include OA (18:1) and 10-HSA ( h 18:0). (D) MS2 spectra with major fragmentation labels for the 10-HSA ( h 18:0) peak from ΔSaohyA /pLCRIS_00661 cultured with OA ( Figure 3 C, lower right). (E) Universally 13 C-labeled 10-HSA ( 13 C 18 -10-HSA) concentrations in supernatants of L. crispatus , L. gasseri , and L. jensenii cultured for 72 h in NYCIII broth with or without universally 13 C-labeled OA ( 13 C 18 -OA; 3.2 mM). (F) 13 C 18 -10-HSA concentrations in supernatants of L. crispatus and L. iners cultured for 72 h in NYCIII broth with or without 13 C 18 -OA (100 μM, a sublethal concentration for L. iners ). (E and F) The same no-OA controls for media and L. crispatus are shown in (E) and (F). Points represent 3 technical replicates per condition. See also Figures S2 and .

Article Snippet: Statistical significance was determined by unpaired t test using the Bonferroni method to correct for multiple hypothesis testing ( ∗ adjusted p < 0.05). (K) Representative h FA extracted ion chromatograms for human CVL samples with CT1 ( L. crispatus -dominant, top) or CT2 ( L. iners -dominant, bottom) bacterial communities, quantified via a targeted metabolomics approach employing picolylamine-based derivatization., (L) DNA gel of PCR products amplified from the primers flanking either LGAS_1630 ( farE ) or LGAS_1351 ( ohyA9 ) from the L. gasseri ATCC 33323 wild-type (WT) strain and ΔohyA9 and ΔfarE genetic knockout (KO) strains.

Techniques: Functional Assay, Activity Assay, Plasmid Preparation, Expressing, Cell Culture, Labeling, Concentration Assay

Journal: Cell

Article Title: Vaginal Lactobacillus fatty acid response mechanisms reveal a metabolite-targeted strategy for bacterial vaginosis treatment

doi: 10.1016/j.cell.2024.07.029

Figure Lengend Snippet: Detection of enzymatic products from ohyA orthologs and characterization of ohyA9 and farE genetic knockouts in L. gasseri , related to Figures 4 and (A) MS2 spectra with major fragmentation labels for 10-HSA standard (Ambeed, A125712-50MG). (B) Extracted ion chromatograms from supernatants of ΔSaohyA /empty vector, ΔSaohyA /p SaohyA , ΔSaohyA /pLCRIS_00558, and ΔSaohyA /pLCRIS_00661 cultured with LOA for 1 h. Annotated peaks include LOA (18:2) and the detected hydroxyFA ( h 18:1). (C) OhyA9 enzymatic activity reaction diagram with LOA substrate. (D) MS2 spectra with major fragmentation labels for the h18:1 peak from ΔSaohyA /pLCRIS_00661 cultured with LOA (lower right of Figure S4 B), identified as 10-hydroxy-12-octadecenoic acid ( h 18:1). (E) OhyA12 enzymatic activity reaction diagram with LOA substrate. (F) MS2 spectra with major fragmentation labels for the h18:1 peak from ΔSaohyA /pLCRIS_00558 cultured with LOA (lower left of Figure S4 B), identified as 13-hydroxy-9-octadecenoic acid. (G) 13 C 18 -10-HSA concentrations in cell pellets of L. crispatus , L. gasseri , and L. jensenii cultured for 72 h in NYCIII broth with and without 13 C 18 -OA (3.2 mM). The cell pellets are from the same cultures as the supernatants shown in Figure 3 E. (H) 13 C 18 -10-HSA concentrations in cell pellets of L. crispatus and L. iners cultured for 72 h in NYCIII broth with and without 13 C 18 -OA (100 μM, which is a sublethal concentration for L. iners ). The cell pellets are from the same cultures as the supernatants shown in Figure 3 F. (I) Presence of gene functions predicted to encode oleate hydratase ( ohyA ) and putative fatty acid efflux pump ( farE ) activity in long-read sequenced, isolate genomes of the indicated FGT species. Presence of gene functions involved in exogenous fatty acid acquisition and utilization ( fakAB , plsC , plsX , and plsY ) is shown for comparison. (J) Untargeted lipidomics was performed on control (blank) media and spent media supernatants collected from strains of diverse FGT bacteria after 72 h of culture in NYCIII broth. Blank media supplemented with 100 μM OA is shown as a positive control. Changes in concentration of key LCFA metabolites for each bacterial species are shown as the log 10 (fold change) of their median relative abundances compared with control media. The plot depicts median fold change values for 5 technical replicates per condition. Statistical significance was determined by unpaired t test using the Bonferroni method to correct for multiple hypothesis testing ( ∗ adjusted p < 0.05). (K) Representative h FA extracted ion chromatograms for human CVL samples with CT1 ( L. crispatus -dominant, top) or CT2 ( L. iners -dominant, bottom) bacterial communities, quantified via a targeted metabolomics approach employing picolylamine-based derivatization. , (L) DNA gel of PCR products amplified from the primers flanking either LGAS_1630 ( farE ) or LGAS_1351 ( ohyA9 ) from the L. gasseri ATCC 33323 wild-type (WT) strain and ΔohyA9 and ΔfarE genetic knockout (KO) strains. Expected amplicon lengths were 3.9 kb for LGAS_1630 in WT, 2.0 kb for LGAS_1351 in WT, while in-frame gene deletions of LGAS_1351 in ΔohyA9 and of LGAS_1630 in ΔfarE each had expected amplicon lengths of 1.2 kb. KO strains were additionally WGS verified. (M) Cultures of L. gasseri WT, L. gasseri Δ farE , and L. gasseri Δ farE /p farE were grown to mid- to late-log phase and exposed to varying concentrations of OA, then ATP release assays were performed. (N) Detection of 13 C-labeled OA ( 13 C 18 -OA) in pellets of L. gasseri Δ farE and Δ farE /p farE genetic mutant strains treated with 100 and 400 μM 13 C 18 -OA in NYCIII broth for 15 min. Pellets were washed two times with ice-cold PBS before sample preparation for targeted 13 C 18 -OA detection. The fold change of [ 13 C 18 -OA] was calculated by dividing the relative abundance of 13 C 18 -OA detected in the 400 μM 13 C 18 -OA condition for each technical replicate by the median relative abundance of 13 C 18 -OA detected in 100 μM 13 C 18 -OA condition. Significance of the difference in fold change was determined by unpaired t test ( ∗∗∗ p < 0.001). Points represent 5 technical replicates per condition. (O) 13 C 18 -10-HSA relative abundance in cell pellets from L. gasseri WT, ΔohyA9 , ΔohyA9 /p ohyA9 , ΔfarE , and ΔfarE /p farE cultured for 24 h in NYCIII broth with and without 13 C 18 -OA (100 μM). Pellets are from the same cultures as the supernatants in Figure 5 D. Points represent 2 or 3 technical replicates per condition. (G and H) Points represent 3 technical replicates per condition.

Article Snippet: Statistical significance was determined by unpaired t test using the Bonferroni method to correct for multiple hypothesis testing ( ∗ adjusted p < 0.05). (K) Representative h FA extracted ion chromatograms for human CVL samples with CT1 ( L. crispatus -dominant, top) or CT2 ( L. iners -dominant, bottom) bacterial communities, quantified via a targeted metabolomics approach employing picolylamine-based derivatization., (L) DNA gel of PCR products amplified from the primers flanking either LGAS_1630 ( farE ) or LGAS_1351 ( ohyA9 ) from the L. gasseri ATCC 33323 wild-type (WT) strain and ΔohyA9 and ΔfarE genetic knockout (KO) strains.

Techniques: Plasmid Preparation, Cell Culture, Activity Assay, Concentration Assay, Comparison, Control, Bacteria, Positive Control, Amplification, Knock-Out, Labeling, Mutagenesis, Sample Prep

Journal: Cell

Article Title: Vaginal Lactobacillus fatty acid response mechanisms reveal a metabolite-targeted strategy for bacterial vaginosis treatment

doi: 10.1016/j.cell.2024.07.029

Figure Lengend Snippet: farE is required for resistance to OA inhibition, and ohyA9 is required for 10-HSA production (A) Relative growth of L. gasseri ATCC 33323 wild-type (WT) and mutant strains, including knockouts of ohyA9 ( ΔohyA9 ) and farE ( ΔfarE ), and ΔfarE complemented with plasmid-overexpressed farE ( ΔfarE /p farE ), cultured in MRS + CQ broth supplemented with varying OA concentrations. (B) Relative growth rescue of L. gasseri WT and mutant strains in lipid-depleted MRS + CQ broth supplemented with varying OA concentrations. (C) MBC assay results for L. gasseri WT and mutant strains in MRS + CQ broth. (D) 13 C 18 -10-HSA relative concentrations in blank media and supernatants from L. gasseri WT, ΔohyA9 , ΔohyA9 complemented with plasmid-overexpressed ohyA9 ( ΔohyA9 /p ohyA9 ), ΔfarE , and ΔfarE /p farE cultured for 24 h in NYCIII broth with or without sub-inhibitory concentrations of 13 C 18 -OA (100 μM). (E) Schematic depicting a proposed model for FarE and OhyA9 activity on exogenous cis -9-uLCFAs in non- iners FGT Lactobacillus species. (A–D) Points represent 2–3 technical replicates per condition. (A–C) Results are representative of ≥2 independent experiments. See also Figure S4 .

Article Snippet: Statistical significance was determined by unpaired t test using the Bonferroni method to correct for multiple hypothesis testing ( ∗ adjusted p < 0.05). (K) Representative h FA extracted ion chromatograms for human CVL samples with CT1 ( L. crispatus -dominant, top) or CT2 ( L. iners -dominant, bottom) bacterial communities, quantified via a targeted metabolomics approach employing picolylamine-based derivatization., (L) DNA gel of PCR products amplified from the primers flanking either LGAS_1630 ( farE ) or LGAS_1351 ( ohyA9 ) from the L. gasseri ATCC 33323 wild-type (WT) strain and ΔohyA9 and ΔfarE genetic knockout (KO) strains.

Techniques: Inhibition, Mutagenesis, Plasmid Preparation, Cell Culture, Activity Assay

Journal: Cell

Article Title: Vaginal Lactobacillus fatty acid response mechanisms reveal a metabolite-targeted strategy for bacterial vaginosis treatment

doi: 10.1016/j.cell.2024.07.029

Figure Lengend Snippet: FGT lactobacilli are FA auxotrophs that exploit OA and its OhyA9-dependent derivative 10-HSA for phospholipid synthesis (A) Fatty acid synthesis II (FASII) and phospholipid synthesis pathways, annotated with predicted gene function presence in FGT Lactobacillus genomes. Predicted gene functions are annotated as missing in a species if absent in >50% of genomes. (B) Growth rescue of L. crispatus ( n = 19), L. gasseri ( n = 3), L. iners ( n = 13), L. jensenii ( n = 8), and L. mulieris ( n = 5) strains in lipid-depleted MRS + CQ broth supplemented with acetate or OA (3.2 mM each), cultured for 72 h. (C) Phosphatidylglycerol (PG) profiles in cell pellets of L. crispatus (top) and L. iners (bottom) cultured for 72 h in NYCIII broth with no added OA (left) or supplemented with 100 μM (middle) or 3.2 mM (right, L. crispatus only) unlabeled OA (black) or 13 C 18 -OA (red). Plots depict representative MS1 spectra. Predominant unlabeled isotopologs of major PG species (black) and the differences in mass/charge (m/z) ratio of their corresponding 13 C-labeled isotopologs (red) are annotated. (D) Growth rescue of non- iners FGT Lactobacillus ( n = 17) and L. iners ( n = 5) strains in lipid-depleted MRS + CQ broth supplemented with varying concentrations of OA (left) or 10-HSA (right), cultured for 72 h. (E) Growth rescue of L. gasseri ATCC 33323 WT and mutant strains in lipid-depleted MRS + CQ broth supplemented with varying concentrations of 10-HSA, cultured for 24 h. (F) Detection of PG lipids in pellets from L. gasseri WT (top), ΔohyA9 (middle), and ΔohyA9 /p ohyA9 (bottom), cultured for 24 h in lipid-depleted MRS + CQ broth containing 50 μM 13 C 18 -OA with (right) or without (left) 400 μM unlabeled 10-HSA (growth shown in Figure S5 G). Plots depict representative MS1 spectra with major 13 C-labeled (black) and partially labeled or unlabeled (red) PG species annotated. (G) L. gasseri genetic mutants strains, Δ ohyA9 and Δ ohyA9 /p ohyA9 , were co-cultured in lipid-depleted MRS + CQ broth supplemented with varying concentrations of OA and 10-HSA, but no erythromycin selection. After 18 h, the ratio of CFU on MRS agar plates with and without erythromycin (respectively representing the Δ ohyA9 /p ohyA9 CFU relative to the total CFU) and relative growth ( Figure S5 I) were determined. The dotted line represents the input CFU ratio. (B, D, and E) Relative growth rescue was calculated as growth relative to median OD600 measurement in non-lipid-depleted MRS + CQ broth. (B and D) Points represent median relative growth for 3 technical replicates per condition. Boxplots represent the 25 th and 75 th percentiles (lower and upper boundaries of boxes, respectively), the median (middle horizontal line), and measurements that fall within 1.5 times the IQR (whiskers). (E) Points represent 3 technical replicates per condition and are representative of ≥2 independent experiments. See also Figure S5 .

Article Snippet: Statistical significance was determined by unpaired t test using the Bonferroni method to correct for multiple hypothesis testing ( ∗ adjusted p < 0.05). (K) Representative h FA extracted ion chromatograms for human CVL samples with CT1 ( L. crispatus -dominant, top) or CT2 ( L. iners -dominant, bottom) bacterial communities, quantified via a targeted metabolomics approach employing picolylamine-based derivatization., (L) DNA gel of PCR products amplified from the primers flanking either LGAS_1630 ( farE ) or LGAS_1351 ( ohyA9 ) from the L. gasseri ATCC 33323 wild-type (WT) strain and ΔohyA9 and ΔfarE genetic knockout (KO) strains.

Techniques: Cell Culture, Labeling, Mutagenesis, Selection

Journal: Cell

Article Title: Vaginal Lactobacillus fatty acid response mechanisms reveal a metabolite-targeted strategy for bacterial vaginosis treatment

doi: 10.1016/j.cell.2024.07.029

Figure Lengend Snippet: Genomic analysis of FASII pathway in FGT Lactobacillus genomes, OA isotope tracing in cultured FGT lactobacilli, and 10-HSA growth effects, related to Figure 6 (A) Presence of gene functions predicted to encode FASII pathway genes in isolate genomes and MAGs of the indicated FGT Lactobacillus species ( n = 1,167). (B) Relative growth of diverse L. crispatus ( n = 19), L. gasseri ( n = 3), L. iners ( n = 13), L. jensenii ( n = 8), and L. mulieris ( n = 5) strains in MRS + CQ broth supplemented with 3.2 mM acetate or 3.2 mM OA. Growth was measured by OD600 after 72 h of culture. (C) Heatmap representing the median incorporation ratio of 13 C 18 -OA in detected diglycerides and central metabolites involved in the tricarboxylic acid (TCA) cycle in cell pellets from representative strains of FGT Lactobacillus species. Bacteria were cultured for 72 h in NYCIII broth with 3.2 mM (top) or 100 μM (bottom). 13 C 18 -OA incorporation ratio was calculated as the signal from the detected 13 C-labeled metabolite relative to the signal of the detected unlabeled metabolite. (D) Heatmap representing the normalized signal of detected unlabeled and labeled phosphatidylglycerol in cell pellets from representative strains of FGT Lactobacillus species. Bacteria were cultured for 72 h in NYCIII broth with 3.2 mM unlabeled OA (left) or 13 C 18 -OA (right). Each row labeled A, B, and C represents a replicate culture of the indicated condition. (E) Relative growth of L. gasseri WT and mutant strains in MRS + CQ broth supplemented with varying concentrations of 10-HSA. Growth was measured by OD600 after 24 h of culture. (F) Relative growth of L. crispatus ( n = 3) and L. iners ( n = 4) strains in MRS + CQ broth supplemented with varying concentrations of OA (left) or 10-HSA (right). Growth was measured by OD600 after 72 h of culture. (G) L. gasseri genetic mutant strains (WT, ΔohyA9 , and ΔohyA9 /p ohyA9 ) were grown for 24 h in lipid-depleted MRS + CQ broth supplemented with 13 C 18 -OA concentration (50 μM) alone and in combination with unlabeled 10-HSA (400 μM, corresponding to the isotopic tracing data in Figure 6 F). Relative growth was calculated relative to the median OD600 measurement in non-lipid-depleted MRS + CQ broth. (H) L. gasseri Δ ohyA9 /p ohyA9 was grown in lipid-depleted MRS + CQ broth supplemented with varying concentrations of OA and 10-HSA, but without erythromycin selection (the p ohyA9 plasmid encodes erythromycin resistance). After 18 h of culture, the ratio of CFU on MRS agar plates with and without erythromycin (respectively representing the amount of the viable Δ ohyA9 /p ohyA9 strain relative to total bacteria) was determined. (I) Growth (determined by OD600) of L. gasseri genetic mutants strains Δ ohyA9 and Δ ohyA9 /p ohyA9 grown in mono-culture or mixed and co-cultured in lipid-depleted MRS + CQ broth supplemented with varying concentrations of OA and 10-HSA, without erythromycin selection. Relative growth was calculated relative to the median OD600 measurement in the 100 μM HSA: 100 μM OA condition. The cultures correspond to the experiment shown in Figure 6 G. (B, E, and F) Relative growth was calculated relative to the median OD600 measurement in the no supplementation control. (B and E) Boxplots represent the 25 th and 75 th percentiles (lower and upper boundaries of boxes, respectively), the median (middle horizontal line), and measurements that fall within 1.5 times the IQR (whiskers). (B and F) Points represent the median of 3 technical replicates per condition. (E) Points represent 3 technical replicates per condition.

Article Snippet: Statistical significance was determined by unpaired t test using the Bonferroni method to correct for multiple hypothesis testing ( ∗ adjusted p < 0.05). (K) Representative h FA extracted ion chromatograms for human CVL samples with CT1 ( L. crispatus -dominant, top) or CT2 ( L. iners -dominant, bottom) bacterial communities, quantified via a targeted metabolomics approach employing picolylamine-based derivatization., (L) DNA gel of PCR products amplified from the primers flanking either LGAS_1630 ( farE ) or LGAS_1351 ( ohyA9 ) from the L. gasseri ATCC 33323 wild-type (WT) strain and ΔohyA9 and ΔfarE genetic knockout (KO) strains.

Techniques: Cell Culture, Bacteria, Labeling, Mutagenesis, Concentration Assay, Selection, Plasmid Preparation, Control

Journal: Journal of Oral Microbiology

Article Title: The role of Lactobacillus plantarum in oral health: a review of current studies

doi: 10.1080/20002297.2024.2411815

Figure Lengend Snippet: Experimental evidence of L. plantarum ’s efficacy in caries control.

Article Snippet: ATCC 10,012 [ ] , CFS , In vitro , Laboratory strain , Planktonic & mono-species biofilm , / , S. mutans ATCC 25175 & S. sobrinus ATCC 33478 , , , , / , The 10012 CSC inhibited the growth of S. mutans and S. sobrinus. , The 10012 CSC significantly reduced biofilm formation by S. mutans and S. sobrinus. , The 10,012 CSC showed better inhibitory abilities than Lactobacillus johnsonii JCM 1022, L. rhamnosus ATCC 7469 and Lactobacillus kefiranofaciens DD2, DD5 and DD6..

Techniques: Control, Sampling, In Vivo, In Vitro, Inhibition, Bacteria, Concentration Assay, Competitive Binding Assay, Modification, Expressing, Activity Assay, Cell Surface Hydrophobicity, Negative Control, Disruption, Ex Vivo, Produced

Journal: Journal of Oral Microbiology

Article Title: The role of Lactobacillus plantarum in oral health: a review of current studies

doi: 10.1080/20002297.2024.2411815

Figure Lengend Snippet: Experimental evidence of L. plantarum ’s efficacy in caries control.

Article Snippet: ATCC 14917 [ ] , CFS , In vitro , Laboratory strain , mono-species biofilm , / , S. mutans UA159 , / , / , 1. The antibiofilm agent, named 1-1-4-3 is a mixture of lactic acid (LA) and valine. , 1. The CFS of L. plantarum showed the strongest antibiofilm activity among the tested CFSs of Lactobacillus casei ATCC 393, Lactobacillus gasseri ATCC 33,323, Lactobacillus fermentum ATCC 14,931 and L. salivarius ATCC 11741.

Techniques: Control, Sampling, In Vivo, In Vitro, Inhibition, Bacteria, Concentration Assay, Competitive Binding Assay, Modification, Expressing, Activity Assay, Cell Surface Hydrophobicity, Negative Control, Disruption, Ex Vivo, Produced

Journal: Journal of Oral Microbiology

Article Title: The role of Lactobacillus plantarum in oral health: a review of current studies

doi: 10.1080/20002297.2024.2411815

Figure Lengend Snippet: Experimental evidence of L. plantarum ’s efficacy in caries control.

Article Snippet: ATCC 8014, ATCC 14,917 [ ] , WBC and plantaricin , In vitro , Clinical isolates , Planktonic and duo-species biofilm , Not mentioned , S. mutans UA159 and C. albicans SC5314 , , , / , , 1. The 14,917 Inhibited the growth of S. mutans and C. albicans clinical isolates. , The 14,917 inhibited the biofilm formation with a compromised biofilm structure with a significantly smaller microbial and extracellular matrix and a less virulent microcolony structure. , 1. The 14917 exhibited better inhibitory effects than L. salivarius 11,741 and L. plantarum 8014..

Techniques: Control, Sampling, In Vivo, In Vitro, Inhibition, Bacteria, Concentration Assay, Competitive Binding Assay, Modification, Expressing, Activity Assay, Cell Surface Hydrophobicity, Negative Control, Disruption, Ex Vivo, Produced