l. monocytogenes Search Results


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  • 99
    ATCC l monocytogenes
    Behaviour of Listeria <t>monocytogenes</t> (Log CFUg -–1 ) during cheese ripening and storage in Pecorino Umbro cheese (data represent the average values±standard deviation of three replicates samples for three cheesemaking replicates).
    L Monocytogenes, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 819 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    89
    GeneTex l monocytogenes
    L. <t>monocytogenes</t> induces local degranulation of primary granules. Neutrophils were incubated with L. monocytogenes ( Lm ) for 1 min, washed, and incubated for a total of 3, 5, 10, and 30 min at 37°C in 10% DS/HBSS + . Cells were fixed, and Lm and CD63 were then fluorescently labeled in nonpermeabilized and permeabilized cells to distinguish granule fusion with the plasma membrane from fusion with sealed phagosomes, respectively. ( A ) Representative images of extracellular ( two top panels ) and intracellular ( two bottom panels ) Lm colocalizing with CD63 at 3 and 10 min. ( B ) MFI ± SEM of CD63 on nonpermeabilized neutrophils was measured by quantitative fluorescence microscopy. Data from a representative experiment, of two, are shown.
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    Xenogen Corporation l monocytogenes
    Rescue of the elektra phenotype by BAC transgenesis Transgenic mice were produced by microinjecting a BAC clone containing the wild-type Slfn2 sequence into single-cell embryos homozygous for the elektra mutation. (a) DNA sequence of the BAC clone and genomic DNA from an elektra homozygote in the region of the Slfn2 elektra mutation. The BAC clone contains the wild-type Slfn2 sequence. (b) Flow cytometric analysis of CD8 and CD4 staining of blood from a WT mouse, a homozygous elektra transgenic mouse carrying the Slfn2 transgene ( elektra-BAC Tg ), and a littermate lacking the transgene ( elektra ). Results are representative of 4 mice per genotype. (c) Survival curves for transgenic mice ( elektra-BAC Tg , n =7), littermates without the transgene ( elektra , n =4), and WT mice ( n =5) upon challenge with 2 × 10 5 PFU of MCMV. (d) Survival curves for transgenic mice ( elektra-BAC Tg , n =7), littermates without the transgene ( elektra , n =5), and WT mice ( n =7) upon challenge with 5 × 10 5 CFU of L. <t>monocytogenes</t> .
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    93
    denka seiken l monocytogenes
    ApaI/AscI PFGE profiles of L. <t>monocytogenes</t> strains 4b and IVb-v1. The dendrogram was calculated and drawn using Bionumerics software. NSW; New South Wales, VIC; Victoria, CA; California, CT; Connecticut, and NK;Not known.
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    97
    Abcam l monocytogenes
    Effect of L. <t>monocytogenes</t> LipA on virulence in mice. (A) Groups of 4 to 5 C57BL/6 wt mice were infected intraperitoneally with 2 × 10 6 L. monocytogenes LO28 wt and Δ lipA bacteria. At days 1 and 3, the spleen and liver were homogenized
    L Monocytogenes, supplied by Abcam, used in various techniques. Bioz Stars score: 97/100, based on 51 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    InvivoGen l monocytogenes
    Effect of L. <t>monocytogenes</t> LipA on virulence in mice. (A) Groups of 4 to 5 C57BL/6 wt mice were infected intraperitoneally with 2 × 10 6 L. monocytogenes LO28 wt and Δ lipA bacteria. At days 1 and 3, the spleen and liver were homogenized
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    Difco l monocytogenes
    Combined PFGE-cluster analysis of L. <t>monocytogenes</t> isolated from serosa and silage samples
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    Thermo Fisher l monocytogenes
    PLS-DA scatter plots of L. <t>monocytogenes</t> inoculated in enriched selective media containing beef sample. (A) PLS-DA Score scatter plot ( R 2 X = 91.5%, R 2 Y = 100%, Q 2 = 91.7%). Green circles (•) indicate data obtained from control beef samples and red stars (★) indicate data from spiked beef samples after 24 h of incubation for L. monocytogenes . The PLS-DA ellipse (solid line) represents the 95% confidence interval. (B) PLS-DA Loading scatter plot.
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    Becton Dickinson l monocytogenes antiserum
    Monocytes and neutrophils are recruited to sites of L. <t>monocytogenes</t> infection
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    PerkinElmer l monocytogenes xen32
    Listeria <t>monocytogenes</t> <t>Xen32</t> strains show attenuated virulence in vivo. BALB/cJ mice (n = 10) were intragastrically inoculated with 1 × 10 10 CFU Lmo-Xen32, Lmo-Xen32-mur, Lmo-EGDe-lux or Lmo-EGDe-mur-lux. (A) Serial BLI of 1 representative mouse from one hour until 7 days post infection as described in Methods (see also Additional file 1 : Figure S1 for reference). Colour bar indicates emitted light with 3 or 4 min integration time in photons/s/cm2/sr. un = uninfected control. (B) Survival rates of BALB/cJ mice (n = 10) after infection with 1 × 10 10 CFU Lmo-Xen32, Lmo-Xen32-mur, Lmo-EGDe-lux or Lmo-EGDe-mur-lux. (C) Body weight loss of BALB/cJ mice infected with the same listerial strains and dosage as shown in (A) and (B) . Graphs demonstrated mean in percent (n = 10) with standard error. Statistical significance between murinised (green stars) and non-murinised (yellow stars) Listeria strains are indicated (*p
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    94
    Bio-Rad l monocytogenes
    Dynamic of thermal inactivation of L. <t>monocytogenes</t> during traditional cooking process of a naturally contaminated kebab.
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    bioMerieux l monocytogenes
    Locus gtcA . (A) Schematic organization of the gtcA region. (Upper part) rpmE-rho region of B. subtilis . The rpmE is shaded in dark gray; the rho is shaded in light gray. (Middle part) rpmE-gtcA-rho region of L. <t>monocytogenes</t> serotype 4b. (Lower part) rpmE-csbB-gtcA-rho region of L. monocytogenes ). Similar residues are shaded, and identical residues are boxed (thick lines). The numbers refer to amino acid length.
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    Eppendorf AG l monocytogenes
    Relative transcript expression profiles of select genes from weakly-adherent (CW35) and strongly-adherent (99-38) strains of L. <t>monocytogenes</t> . Panel A, from cells recovered from planktonic growth at 30 °C. Panel B, from cells attached to glass beads during growth at 30 °C. Panel C, from planktonic cells grown at 42 °C. Expression is relative to that of the reference gene, 16S rRNA. All data bars represent the means of triplicate replications for gene expression RT-qPCR assays. Error bars indicate the standard deviation from the mean. Expression was normalized (×10 7 factor) to eliminate negative expression levels.
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    Lab M Ltd l monocytogenes
    Relative transcript expression profiles of select genes from weakly-adherent (CW35) and strongly-adherent (99-38) strains of L. <t>monocytogenes</t> . Panel A, from cells recovered from planktonic growth at 30 °C. Panel B, from cells attached to glass beads during growth at 30 °C. Panel C, from planktonic cells grown at 42 °C. Expression is relative to that of the reference gene, 16S rRNA. All data bars represent the means of triplicate replications for gene expression RT-qPCR assays. Error bars indicate the standard deviation from the mean. Expression was normalized (×10 7 factor) to eliminate negative expression levels.
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    Lista International l monocytogenes
    Relative transcript expression profiles of select genes from weakly-adherent (CW35) and strongly-adherent (99-38) strains of L. <t>monocytogenes</t> . Panel A, from cells recovered from planktonic growth at 30 °C. Panel B, from cells attached to glass beads during growth at 30 °C. Panel C, from planktonic cells grown at 42 °C. Expression is relative to that of the reference gene, 16S rRNA. All data bars represent the means of triplicate replications for gene expression RT-qPCR assays. Error bars indicate the standard deviation from the mean. Expression was normalized (×10 7 factor) to eliminate negative expression levels.
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    Medical Diagnostic Laboratories LLC l monocytogenes
    Relative transcript expression profiles of select genes from weakly-adherent (CW35) and strongly-adherent (99-38) strains of L. <t>monocytogenes</t> . Panel A, from cells recovered from planktonic growth at 30 °C. Panel B, from cells attached to glass beads during growth at 30 °C. Panel C, from planktonic cells grown at 42 °C. Expression is relative to that of the reference gene, 16S rRNA. All data bars represent the means of triplicate replications for gene expression RT-qPCR assays. Error bars indicate the standard deviation from the mean. Expression was normalized (×10 7 factor) to eliminate negative expression levels.
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    Biosynth l monocytogenes
    ELISA O-antigen and H-antigen reactions of serogroup 1/2 and 3 L. <t>monocytogenes</t> strains. (A) Strain J0098, serotype 1/2a; (B) strain G848, serotype 1/2b; (C) strain G-3321, serotype 1/2c; (D) strain J0095, serotype 3a; (E) strain cpp81, serotype 3b; (F) strain J0096, serotype 3c.
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    Merck & Co l monocytogenes
    Recretion of selected InlA mutations in EGD-e . A . Comparison of the invasion attributes of EGD-e and EGD-e InlA m * (Ser192Asn/Trp369Ser). Exponential phase L. <t>monocytogenes</t> cells (OD = 0.8) were invaded (MOI of 25:1) in triplicate for 1 h before overlaying with gentamicin. Invasion was expressed as the average cfu count per well (with standard deviation) or invasion relative to EGD-e (below graph). The graph is representative of the data from three independent experiments. B . The relative virulence of EGD-e compared against EGD-e InlA m * (tagged with pIMC3 kan and pIMC3 ery respectively) was accessed by competitive index after i.v. infection (1 × 10 4 cfu of each strain) of 15 Balb/c mice. On each subsequent day 5 mice were euthanized and spleens and livers aseptically removed and enumerated. Data are presented as the mean and standard deviation of 5 mice, competitive indices and statistical analysis was conducted using the one sample t test as described previously [ 18 ]. NS = Not significant. C . Oral inoculations of Balb/c mice with EGD-e::pIMC3 kan and EGD-e InlA m * ::pIMC ery mixed at a 1:1 ratio in a total inoculum of 1 × 10 10 cfu/200 μl containing 100 mg of CaCO 3 . *** = p
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    Merck KGaA l monocytogenes
    Influence of pG, GO, and rGO on the growth of Listeria <t>monocytogenes</t> and Salmonella enterica at 25 and 250 μg/mL. Data presented are the average of triplicate determinations, with error bars representing mean standard error.
    L Monocytogenes, supplied by Merck KGaA, used in various techniques. Bioz Stars score: 93/100, based on 55 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    SAS institute l monocytogenes
    Influence of pG, GO, and rGO on the growth of Listeria <t>monocytogenes</t> and Salmonella enterica at 25 and 250 μg/mL. Data presented are the average of triplicate determinations, with error bars representing mean standard error.
    L Monocytogenes, supplied by SAS institute, used in various techniques. Bioz Stars score: 92/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Luminex l monocytogenes
    In IL-17 Receptor Deficient Mice, M. Pulmonis Infection Does Not Facilitate Clearance of L. <t>Monocytogenes</t> or Increase Gr-1+CD11b+ Cells. At day 17 p.i. with M. pulmonis or broth, C57BL/6 and IL-17 receptor deficient mice were infected with L. monocytogenes
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    Millipore l monocytogenes
    Schematic outlining the interaction of L. <t>monocytogenes</t> with bile during infection. (A) L. monocytogenes ). (B) Infection of the gallbladder is mediated by an as-yet-undetermined mechanism. Growth in GB bile within the lumen requires specific metabolic processes but is independent of specific resistance mechanisms (the present study).
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    Antibodies-Online Inc l monocytogenes
    Schematic outlining the interaction of L. <t>monocytogenes</t> with bile during infection. (A) L. monocytogenes ). (B) Infection of the gallbladder is mediated by an as-yet-undetermined mechanism. Growth in GB bile within the lumen requires specific metabolic processes but is independent of specific resistance mechanisms (the present study).
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    Eurofins l monocytogenes
    Fluorescence properties of permeabilized L. <t>monocytogenes</t> EGD-e/pNZ-P help -pHluorin bacteria. (A) In order to determine the appropriate concentration for permeabilization, L. monocytogenes EGD-e was incubated with different concentrations of CTAB (0 - 0.01 %) and stained with Syto ® 9 and propidium iodide (PI) to discriminate intact, live from permeabilized, dead bacteria. Images in the Syto ® 9 and PI channels were aquired with a 100x objective and appropriate filter sets for excitation and emission of the two dyes (Scale bars: 2 μm). (B) L. monocytogenes EGD-e/pNZ-P help -pHluorin was permeabilized in LMB at different pH containing 0.005% CTAB and, at the indicated pH, excitation spectra (350–490 nm) was aquired measuring relative fluorescence intensity (RFI) at 510 nm emission.
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    Fisher Scientific l monocytogenes
    Mean lag-phase duration (h), maximum growth rate (ΔOD 600 /h), and maximum OD 600 values for L. <t>monocytogenes</t> isolates possessing LGI1 ( n = 9) and isolates without LGI1 ( n = 8) grown in the presence of sublethal concentrations of E-San (1.6 μl/ml)
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    DSMZ l monocytogenes
    (A) 5′-Nuclease PCR analysis of serial 10-fold dilutions of L. <t>monocytogenes</t> DNA. C T values are plotted against the calculated CFU (i.e., 10-fold dilutions of the bacterial DNA from 3.125 × 10 6 CFU/μl). The straight line, which is calculated by linear regression ( y = −3.56 x [CFU] + 40.7) shows a square regression coefficient of R 2 = 0.993. The standard deviations based on three PCR reactions are indicated. (B) 5′-Nuclease PCR analysis of serial 10-fold dilutions of L. monocytogenes cells. C T values plotted against the number of CFU of L. monocytogenes . Template DNA was extracted from samples of cells containing serial 10-fold dilutions of approximately (5.0 ± 0.3) × 10 7 CFU of L. monocytogenes . The straight line, which is calculated by linear regression ( y = −4.12 x [CFU] + 45.5) shows a square regression coefficient of R 2 = 0.995. The standard deviations based on three PCR reactions are indicated.
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    MatTek l monocytogenes
    Effect of pH i on actin polymerization-driven motility of L. <t>monocytogenes</t> . A , schematic of NHE1 and the point mutation that blocks ion translocation of NHE1 while maintaining its ability to interact with adaptor proteins through its long cytoplasmic tail
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    Pasteur Institute l monocytogenes
    Increased bacterial loads after infection of TLR2 −/− mice with L. <t>monocytogenes</t> . TLR2 +/+ (black bars), TLR2 −/− (white bars), and MyD88 −/− (hatched bars) mice were infected with 2 × 10 5 CFU of Listeria i.v. and sacrificed at day 2 postinfection, and the total numbers of CFU per liver (A) and spleen (B) were determined. Results are from one representative experiment of two independent experiments. Results are expressed as means ± SD. *, P
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    Vitek Inc l monocytogenes
    Growth of L. <t>monocytogenes</t> on cooked, modified-atmosphere, hot-packed poultry injected with regular brine (open symbols) or test brine containing sodium lactate and ALTA 2341 (solid symbols) and stored at 3.5°C (▴), 6.5°C (•), or 10°C (■). Counts below 1,000 CFU/piece were determined by MPN.
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    Bio-Synthesis Inc l monocytogenes
    Restriction-modification system genes in the L. <t>monocytogenes</t> serotype 4b genome. Annotation and visualization of the region were done using GAMOLA and ARTEMIS as described in Materials and Methods. Putative open reading frames designated 5-methyl cytosine restriction, methylase, and putative Sau3AI correspond to 85R, 85M, and 85S, respectively, described in the text. Black bars indicate probes used for Southern blot-based detection of 85R, 85M, and 85S. Arrows indicate direction of transcription. The deduced G+C contents of the genes are shown at the top.
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    85
    Biotechnology Information l monocytogenes
    The AgDI pathway and organization of the gene cluster in L. <t>monocytogenes</t> . A , the AgDI pathway. Agmatine is first hydrolyzed to form N -carbamoylputrescine by AgDI. N -Carbamoylputrescine is then converted into putrescine and carbamoylphosphate by putrescine
    L Monocytogenes, supplied by Biotechnology Information, used in various techniques. Bioz Stars score: 85/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    ATUM l monocytogenes
    L. <t>monocytogenes</t> (LM) is trapped in late endosomes and acidified lysosomes in EVT. ( A ) Percent co-localization of early endosomal marker Rab5 and autophagosomal marker LC3 with LM. ( B ) Percent co-localization of late endosomal marker Rab7, lysosomal marker Lamp1 and acidotropic dye Lysotracker (LT) with LM. p.i. = post-inoculation. Each data point is an average of 3 independent experiments. Bars represent SEM. ( C–E ) Representative images of GFP-expressing LM (green) co-localizing with markers Rab7, Lamp1, and LT (red). The color-merged images also show DAPI counterstain (white). Scale bars are 10 µm.
    L Monocytogenes, supplied by ATUM, used in various techniques. Bioz Stars score: 93/100, based on 40 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Behaviour of Listeria monocytogenes (Log CFUg -–1 ) during cheese ripening and storage in Pecorino Umbro cheese (data represent the average values±standard deviation of three replicates samples for three cheesemaking replicates).

    Journal: Italian Journal of Food Safety

    Article Title: Behaviour of Listeria Monocytogenes in Artisanal Raw Milk Pecorino Umbro Cheese: A Microbiological Challenge Test

    doi: 10.4081/ijfs.2015.5370

    Figure Lengend Snippet: Behaviour of Listeria monocytogenes (Log CFUg -–1 ) during cheese ripening and storage in Pecorino Umbro cheese (data represent the average values±standard deviation of three replicates samples for three cheesemaking replicates).

    Article Snippet: The presence of a certain % of NaCl was one method used to control the growth of L. monocytogenes , but the concentration used in the experiment is not demonstrated to limit L. monocytogenes growth (Faleiro et al ., 2003).

    Techniques: Standard Deviation

    Agarose gel electrophoresis showing monoplex and multiplex PCR-amplified products. M, 100-bp DNA marker; lane 1 , Salmonella enteritidis ATCC13076; lane 2 , L. monocytogenes ATCC19111; lane 3 , E. coli O157:H7 (HCMUS); lane 4 , 16S rRNA of bacterial DNA; lane 5 , a mixture of the three DNA templates of pathogens before the optimizing; lane 6 , the multiplex PCR-optimized conditions with four targets; and NC, negative control

    Journal: 3 Biotech

    Article Title: Multiplex PCR for simultaneous identification of E. coli O157:H7, Salmonella spp. and L. monocytogenes in food

    doi: 10.1007/s13205-016-0523-6

    Figure Lengend Snippet: Agarose gel electrophoresis showing monoplex and multiplex PCR-amplified products. M, 100-bp DNA marker; lane 1 , Salmonella enteritidis ATCC13076; lane 2 , L. monocytogenes ATCC19111; lane 3 , E. coli O157:H7 (HCMUS); lane 4 , 16S rRNA of bacterial DNA; lane 5 , a mixture of the three DNA templates of pathogens before the optimizing; lane 6 , the multiplex PCR-optimized conditions with four targets; and NC, negative control

    Article Snippet: E. coli O157:H7 (HCMUS), E. coli O157:H7 (NIHE), E. coli O157:H7 (NLU), S. enteritidis ATCC13076, and L. monocytogenes ATCC19111 that were positive in the multiplex PCR assay, while all non-target strains were negative in the assay and all non-target bacterial including E. coli ATCC 11775, E. coli ATCC 25922, E. coli (Ahmed et al. ), E. coli (Amagliani et al. ), E. coli (Aranda et al. ), E. coli (Cetinkaya et al. ), E. coli (Coffey et al. ), C. perfringens ATCC13124, B. cereus ATCC11778, V. cholerae ATCC17802 and S. sonnei ATCC 9290 were negative in the assay, whereas 16S rRNA was amplified as expected.

    Techniques: Agarose Gel Electrophoresis, Multiplex Assay, Polymerase Chain Reaction, Amplification, Marker, Negative Control

    CTL activity in the spleen as determined ex vivo by 51 Cr release assay. Mice were immunized with L. monocytogenes EGD (□), a combination of L. monocytogenes ATCC 15313 and LLO-liposome (○), ATCC 15313 alone (▵), or LLO-liposome alone (•) for 6 days. Cells obtained from nonimmune mice (■) were used as the control. 51 Cr-labeled target cells (BMMΦ) infected with EGD) were cultured with effector cells form each group of mice at ratios ranging form 80:1 to 10:1 for 4 h, and the percent specific lysis was calculated. Data represent mean counts for three wells.

    Journal: Infection and Immunity

    Article Title: Induction of Protective T Cells against Listeria monocytogenes in Mice by Immunization with a Listeriolysin O-Negative Avirulent Strain of Bacteria and Liposome-Encapsulated Listeriolysin O

    doi:

    Figure Lengend Snippet: CTL activity in the spleen as determined ex vivo by 51 Cr release assay. Mice were immunized with L. monocytogenes EGD (□), a combination of L. monocytogenes ATCC 15313 and LLO-liposome (○), ATCC 15313 alone (▵), or LLO-liposome alone (•) for 6 days. Cells obtained from nonimmune mice (■) were used as the control. 51 Cr-labeled target cells (BMMΦ) infected with EGD) were cultured with effector cells form each group of mice at ratios ranging form 80:1 to 10:1 for 4 h, and the percent specific lysis was calculated. Data represent mean counts for three wells.

    Article Snippet: Mice were immunized intravenously with 2 × 103 CFU of L. monocytogenes EGD, 2 × 107 CFU of L. monocytogenes ATCC 15313, 200 μl of LLO-liposome, or 2 × 107 CFU of L. monocytogenes ATCC 15313 mixed with 200 μl of LLO-liposome.

    Techniques: CTL Assay, Activity Assay, Ex Vivo, Release Assay, Mouse Assay, Labeling, Infection, Cell Culture, Lysis

    Kinetic change in the CFU of L. monocytogenes EGD and ATCC 15313 after intravenous immunization. Mice were immunized with 10 4 CFU of EGD (□) or with 2 × 10 7 CFU of ATCC 15313 alone (•) or with LLO-liposome (200 μl/head) (○). At the time points indicated, CFU counts in the spleen were determined. The hatched area indicates the nondetectable level. Data represent mean counts for three mice ± standard deviation.

    Journal: Infection and Immunity

    Article Title: Induction of Protective T Cells against Listeria monocytogenes in Mice by Immunization with a Listeriolysin O-Negative Avirulent Strain of Bacteria and Liposome-Encapsulated Listeriolysin O

    doi:

    Figure Lengend Snippet: Kinetic change in the CFU of L. monocytogenes EGD and ATCC 15313 after intravenous immunization. Mice were immunized with 10 4 CFU of EGD (□) or with 2 × 10 7 CFU of ATCC 15313 alone (•) or with LLO-liposome (200 μl/head) (○). At the time points indicated, CFU counts in the spleen were determined. The hatched area indicates the nondetectable level. Data represent mean counts for three mice ± standard deviation.

    Article Snippet: Mice were immunized intravenously with 2 × 103 CFU of L. monocytogenes EGD, 2 × 107 CFU of L. monocytogenes ATCC 15313, 200 μl of LLO-liposome, or 2 × 107 CFU of L. monocytogenes ATCC 15313 mixed with 200 μl of LLO-liposome.

    Techniques: Mouse Assay, Standard Deviation

    Protective immunity induced by immunization with a combination of 2 × 10 7 CFU of L. monocytogenes ( L. m. ) ATCC 15313 and LLO-liposome. The level in mice immunized with virulent strain EGD is shown as positive control. Six days after immunization, mice were challenged intravenously with 2 × 10 4 CFU of strain EGD, and the CFU number in the spleen was determined 2 days later. The level of protective immunity was calculated as log 10 CFU in nonimmune control − log 10 CFU in the immune group. Data represent mean counts for three mice ± standard deviation.

    Journal: Infection and Immunity

    Article Title: Induction of Protective T Cells against Listeria monocytogenes in Mice by Immunization with a Listeriolysin O-Negative Avirulent Strain of Bacteria and Liposome-Encapsulated Listeriolysin O

    doi:

    Figure Lengend Snippet: Protective immunity induced by immunization with a combination of 2 × 10 7 CFU of L. monocytogenes ( L. m. ) ATCC 15313 and LLO-liposome. The level in mice immunized with virulent strain EGD is shown as positive control. Six days after immunization, mice were challenged intravenously with 2 × 10 4 CFU of strain EGD, and the CFU number in the spleen was determined 2 days later. The level of protective immunity was calculated as log 10 CFU in nonimmune control − log 10 CFU in the immune group. Data represent mean counts for three mice ± standard deviation.

    Article Snippet: Mice were immunized intravenously with 2 × 103 CFU of L. monocytogenes EGD, 2 × 107 CFU of L. monocytogenes ATCC 15313, 200 μl of LLO-liposome, or 2 × 107 CFU of L. monocytogenes ATCC 15313 mixed with 200 μl of LLO-liposome.

    Techniques: Mouse Assay, Positive Control, Standard Deviation

    Expression of IFN-γ mRNA after immunization with L. monocytogenes ATCC 15313 or EGD as determined by RT-PCR. Mice were immunized with 2 × 10 7 CFU of ATCC 15313 (lane b), ATCC 15313 plus 200 μl of LLO-liposome (lane c), or 2 × 10 3 CFU of EGD (lane d). Lane a, control; lane M, 100 bp ladder. Total cellular RNA was extracted from the spleen 24 h after stimulation for RT-PCR.

    Journal: Infection and Immunity

    Article Title: Induction of Protective T Cells against Listeria monocytogenes in Mice by Immunization with a Listeriolysin O-Negative Avirulent Strain of Bacteria and Liposome-Encapsulated Listeriolysin O

    doi:

    Figure Lengend Snippet: Expression of IFN-γ mRNA after immunization with L. monocytogenes ATCC 15313 or EGD as determined by RT-PCR. Mice were immunized with 2 × 10 7 CFU of ATCC 15313 (lane b), ATCC 15313 plus 200 μl of LLO-liposome (lane c), or 2 × 10 3 CFU of EGD (lane d). Lane a, control; lane M, 100 bp ladder. Total cellular RNA was extracted from the spleen 24 h after stimulation for RT-PCR.

    Article Snippet: Mice were immunized intravenously with 2 × 103 CFU of L. monocytogenes EGD, 2 × 107 CFU of L. monocytogenes ATCC 15313, 200 μl of LLO-liposome, or 2 × 107 CFU of L. monocytogenes ATCC 15313 mixed with 200 μl of LLO-liposome.

    Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Mouse Assay

    Transmission electron micrographs of L. monocytogenes V7 non-adapted (A,C) and oxidative stress adapted cells (B,D) gradually exposing to 375 ppm (3/4 MIC) from 250 ppm (1/2 MIC) of chlorine over 7 days. Micrographs represent planktonic cells: (A,B) control cells at different magnifications; (C,D) elongated cells and bud formation indicated by arrow, and multi-chromosome formation and inhibition of cell division indicated by red circle in chlorine adapted cells at different magnifications.

    Journal: Frontiers in Microbiology

    Article Title: Listeria monocytogenes Response to Sublethal Chlorine Induced Oxidative Stress on Homologous and Heterologous Stress Adaptation

    doi: 10.3389/fmicb.2018.02050

    Figure Lengend Snippet: Transmission electron micrographs of L. monocytogenes V7 non-adapted (A,C) and oxidative stress adapted cells (B,D) gradually exposing to 375 ppm (3/4 MIC) from 250 ppm (1/2 MIC) of chlorine over 7 days. Micrographs represent planktonic cells: (A,B) control cells at different magnifications; (C,D) elongated cells and bud formation indicated by arrow, and multi-chromosome formation and inhibition of cell division indicated by red circle in chlorine adapted cells at different magnifications.

    Article Snippet: L. monocytogenes isolates from meat and dairy have also been found to be more resistant than its reference ATCC 19116 strain.

    Techniques: Transmission Assay, Inhibition

    Transmission electron micrographs of Listeria monocytogenes Scott A non-adapted (A,B) and oxidative stress adapted cells (C,D) after gradually exposing to 375 ppm (3/4 MIC) from 250 ppm (1/2 MIC) of chlorine over 7 days. Micrographs represent planktonic cells: (A,B) control cells at different magnifications; (C,D) elongated cells and bud formation, indicated by arrow and multi-chromosome formation indicated by red circle in chlorine adapted cells at different magnifications.

    Journal: Frontiers in Microbiology

    Article Title: Listeria monocytogenes Response to Sublethal Chlorine Induced Oxidative Stress on Homologous and Heterologous Stress Adaptation

    doi: 10.3389/fmicb.2018.02050

    Figure Lengend Snippet: Transmission electron micrographs of Listeria monocytogenes Scott A non-adapted (A,B) and oxidative stress adapted cells (C,D) after gradually exposing to 375 ppm (3/4 MIC) from 250 ppm (1/2 MIC) of chlorine over 7 days. Micrographs represent planktonic cells: (A,B) control cells at different magnifications; (C,D) elongated cells and bud formation, indicated by arrow and multi-chromosome formation indicated by red circle in chlorine adapted cells at different magnifications.

    Article Snippet: L. monocytogenes isolates from meat and dairy have also been found to be more resistant than its reference ATCC 19116 strain.

    Techniques: Transmission Assay

    Electron micrographs of IFN-γ-activated THP-1 macrophages infected with nonadapted (a and b) and acid-adapted (c and d) L. monocytogenes LM2. A number of nonadapted bacterial cells show dramatic structural damage due to phagosomal digestion (a), whereas only a few cells can be observed free in the cytoplasm (b). Acid-adapted L. monocytogenes cells appear either intact and in active multiplication (arrow) in the phagosome (c) or free in the cytoplasm, with actin tails (d). Bars, 0.5 μm.

    Journal: Infection and Immunity

    Article Title: Effect of Acid Adaptation on the Fate of Listeria monocytogenes in THP-1 Human Macrophages Activated by Gamma Interferon

    doi: 10.1128/IAI.70.8.4369-4378.2002

    Figure Lengend Snippet: Electron micrographs of IFN-γ-activated THP-1 macrophages infected with nonadapted (a and b) and acid-adapted (c and d) L. monocytogenes LM2. A number of nonadapted bacterial cells show dramatic structural damage due to phagosomal digestion (a), whereas only a few cells can be observed free in the cytoplasm (b). Acid-adapted L. monocytogenes cells appear either intact and in active multiplication (arrow) in the phagosome (c) or free in the cytoplasm, with actin tails (d). Bars, 0.5 μm.

    Article Snippet: As an example, a comparison of the acid-resistant phenotypes of L. monocytogenes LM2 and ATCC 7644 is shown in Fig. .

    Techniques: Infection

    RNA slot blot analysis of actA , clpC , gad , hly , inlA , plcA , prfA , and sodC transcripts in nonadapted and acid-adapted L. monocytogenes LM2. Total RNAs were extracted from exponential cultures ( A 600 ≅ 0.4) of L. monocytogenes LM2 in BHI at pH 5.1 and 7.2. The amount of bound probe for each RNA slot was normalized by the amount of bound 16S rRNA probe. The percentage of transcription is relative to that for the non-acid-adapted condition.

    Journal: Infection and Immunity

    Article Title: Effect of Acid Adaptation on the Fate of Listeria monocytogenes in THP-1 Human Macrophages Activated by Gamma Interferon

    doi: 10.1128/IAI.70.8.4369-4378.2002

    Figure Lengend Snippet: RNA slot blot analysis of actA , clpC , gad , hly , inlA , plcA , prfA , and sodC transcripts in nonadapted and acid-adapted L. monocytogenes LM2. Total RNAs were extracted from exponential cultures ( A 600 ≅ 0.4) of L. monocytogenes LM2 in BHI at pH 5.1 and 7.2. The amount of bound probe for each RNA slot was normalized by the amount of bound 16S rRNA probe. The percentage of transcription is relative to that for the non-acid-adapted condition.

    Article Snippet: As an example, a comparison of the acid-resistant phenotypes of L. monocytogenes LM2 and ATCC 7644 is shown in Fig. .

    Techniques: Dot Blot

    ATR in wild-type L. monocytogenes LM2 and ATCC 7644. Survival rates are reported for acid (pH 5.1)-adapted (•) and nonadapted (○) strain LM2 and for acid-adapted (▪) and nonadapted (□) strain ATCC 7644. Bacterial counts were performed at different times of incubation in BHI acidified at pH 3.5. Error bars, SD for five independent experiments.

    Journal: Infection and Immunity

    Article Title: Effect of Acid Adaptation on the Fate of Listeria monocytogenes in THP-1 Human Macrophages Activated by Gamma Interferon

    doi: 10.1128/IAI.70.8.4369-4378.2002

    Figure Lengend Snippet: ATR in wild-type L. monocytogenes LM2 and ATCC 7644. Survival rates are reported for acid (pH 5.1)-adapted (•) and nonadapted (○) strain LM2 and for acid-adapted (▪) and nonadapted (□) strain ATCC 7644. Bacterial counts were performed at different times of incubation in BHI acidified at pH 3.5. Error bars, SD for five independent experiments.

    Article Snippet: As an example, a comparison of the acid-resistant phenotypes of L. monocytogenes LM2 and ATCC 7644 is shown in Fig. .

    Techniques: Incubation

    RI of L. monocytogenes LM2 in THP-1 macrophages. Circles, bacteria in nonactivated cells; triangles, bacteria in IFN-γ-activated cells. Solid symbols, acid-adapted bacteria; open symbols, nonadapted bacteria. Each data point is the mean intracellular L. monocytogenes RI from one experiment representative of six performed.

    Journal: Infection and Immunity

    Article Title: Effect of Acid Adaptation on the Fate of Listeria monocytogenes in THP-1 Human Macrophages Activated by Gamma Interferon

    doi: 10.1128/IAI.70.8.4369-4378.2002

    Figure Lengend Snippet: RI of L. monocytogenes LM2 in THP-1 macrophages. Circles, bacteria in nonactivated cells; triangles, bacteria in IFN-γ-activated cells. Solid symbols, acid-adapted bacteria; open symbols, nonadapted bacteria. Each data point is the mean intracellular L. monocytogenes RI from one experiment representative of six performed.

    Article Snippet: As an example, a comparison of the acid-resistant phenotypes of L. monocytogenes LM2 and ATCC 7644 is shown in Fig. .

    Techniques:

    CTL lysis of J774 cells infected with L. monocytogenes. J774 macrophages were treated with virulent L. monocytogenes (□), avirulent L. monocytogenes (○), heat-killed virulent L. monocytogenes (●) or left untreated (■). Effectors were added to 51 Cr-labelled targets at the ratios indicated and percentage 51 . METHODS. CTL line LmT was obtained from a (BALB/c × C57BL/6) F 1 mouse 8 days after infection with 5 × 10 3 virulent L. monocytogenes (ATCC 43251). Flasks for in vitro stimulation were prepared by adding 5 × 10 6 . After 24 h the flasks were infected with 1 ml virulent L. monocytogenes in mid-log phase ( A 600 of 0.1) for 25 min, at which time the medium was replaced with RP10 containing 5μg ml −1 gentamicin. Three hours later medium was replaced with RP10 containing 100 U ml −1 penicillin, 100 μg ml −1 streptomycin and 50 μg ml −1 gentamicin and incubated overnight. Immune splenocytes (3×10 7 ) were added to the flasks (in 20 ml volume) and incubated upright. CTL were restimulated every 7–12 days and after the first two stimulations the media was suplemented with interleukin-2. CTL were assayed by labelling 10 6 J774 cells with 100 μCi 51 Cr for 45 min in antibiotic-free media. Cells (10 4 ) were placed into wells and allowed to adhere for 30 min. Cells were then exposed to either 2 × 10 6 live or heat-killed (60 °C for 30 min) virulent or avirulent (ATCC 43250) L. monocytogenes for 25 min. Medium was replaced with RP10 containing 5 μg ml −1 gentamicin, CTL were added and lysis was allowed to proceed for 3 h. Spontaneous lysis of all target cells was

    Journal: Nature

    Article Title: Precise prediction of a dominant class I MHC-restricted epitope of Listeria monocytogenes

    doi: 10.1038/353852a0

    Figure Lengend Snippet: CTL lysis of J774 cells infected with L. monocytogenes. J774 macrophages were treated with virulent L. monocytogenes (□), avirulent L. monocytogenes (○), heat-killed virulent L. monocytogenes (●) or left untreated (■). Effectors were added to 51 Cr-labelled targets at the ratios indicated and percentage 51 . METHODS. CTL line LmT was obtained from a (BALB/c × C57BL/6) F 1 mouse 8 days after infection with 5 × 10 3 virulent L. monocytogenes (ATCC 43251). Flasks for in vitro stimulation were prepared by adding 5 × 10 6 . After 24 h the flasks were infected with 1 ml virulent L. monocytogenes in mid-log phase ( A 600 of 0.1) for 25 min, at which time the medium was replaced with RP10 containing 5μg ml −1 gentamicin. Three hours later medium was replaced with RP10 containing 100 U ml −1 penicillin, 100 μg ml −1 streptomycin and 50 μg ml −1 gentamicin and incubated overnight. Immune splenocytes (3×10 7 ) were added to the flasks (in 20 ml volume) and incubated upright. CTL were restimulated every 7–12 days and after the first two stimulations the media was suplemented with interleukin-2. CTL were assayed by labelling 10 6 J774 cells with 100 μCi 51 Cr for 45 min in antibiotic-free media. Cells (10 4 ) were placed into wells and allowed to adhere for 30 min. Cells were then exposed to either 2 × 10 6 live or heat-killed (60 °C for 30 min) virulent or avirulent (ATCC 43250) L. monocytogenes for 25 min. Medium was replaced with RP10 containing 5 μg ml −1 gentamicin, CTL were added and lysis was allowed to proceed for 3 h. Spontaneous lysis of all target cells was

    Article Snippet: J774 cells infected with avirulent L. monocytogenes that do not express LLO (ATCC 43250) or treated with heat-killed virulent L. monocytogenes were not lysed significantly above background ( ).

    Techniques: CTL Assay, Lysis, Infection, In Vitro, Incubation

    Auto-aggregation of Lactococcus lactis subsp. lactis DF04Mi, Listeria monocytogenes 711, Lactobacillus sakei ATCC 15521 and Enterococcus faecalis ATCC 19433 and co-aggregation of Lactococcus lactis subsp. lactis DF04Mi with Listeria monocytogenes 711, Lactobacillus sakei ATCC 15521 or Enterococcus faecalis ATCC 19433.

    Journal: Brazilian Journal of Microbiology

    Article Title: Bacteriocinogenic Lactococcus lactis subsp. lactis DF04Mi isolated from goat milk: Evaluation of the probiotic potential

    doi:

    Figure Lengend Snippet: Auto-aggregation of Lactococcus lactis subsp. lactis DF04Mi, Listeria monocytogenes 711, Lactobacillus sakei ATCC 15521 and Enterococcus faecalis ATCC 19433 and co-aggregation of Lactococcus lactis subsp. lactis DF04Mi with Listeria monocytogenes 711, Lactobacillus sakei ATCC 15521 or Enterococcus faecalis ATCC 19433.

    Article Snippet: Auto-aggregation and co-aggregation with L. monocytogenes 711, E. faecalis ATCC 19433 and Lb. sakei ATCC 15521 The cells of a culture of Lc. lactis DF04Mi grown in MRS broth (Difco) for 24 h at 37 °C were harvested by centrifugation (7,000 × g , 10 min, 20 °C), washed and resuspended in sterile saline (0.85% NaCl, w/v).

    Techniques:

    Results obtained from the multiplex PCR product of bacterial genomics. Lane 1: 100 bp DNA ladder Lane 2: L. monocytogenes (210 bp), C. jejuni (160 bp), B. cereus (300 bp)

    Journal: Iranian Journal of Basic Medical Sciences

    Article Title: Presenting a rapid method for detection of Bacillus cereus, Listeria monocytogenes and Campylobacter jejuni in food samples

    doi: 10.22038/IJBMS.2017.9275

    Figure Lengend Snippet: Results obtained from the multiplex PCR product of bacterial genomics. Lane 1: 100 bp DNA ladder Lane 2: L. monocytogenes (210 bp), C. jejuni (160 bp), B. cereus (300 bp)

    Article Snippet: L. monocytogenes bacterium obtained from Iranian Centre of Industrial and Medical Fungi and Bacteria Collection (PTCC 1298) Following bacteria were prepared from the Iranian Reference Laboratory: Salmonella typhi ATCC 700931, Shigella dysentery ATCC 13313, Yersinia pestis ATCCBAA-1511D-5, Staphylococcus aureus ATCC 25923 , Clostridium perfringes ATCC 13124, Clostridium botulinum ATCC 3502, Vibrio cholera ATCC 9459.

    Techniques: Multiplex Assay, Polymerase Chain Reaction

    Study of the sensitivity of PCR on 1%5 polyacrylamide gel electrophoresis Lane 7: 100 bp DNA marker Lane 1-6: PCR products with the different dilutions of L. monocytogenes, B. cereus and C . jejuni with 10 1 -10 6

    Journal: Iranian Journal of Basic Medical Sciences

    Article Title: Presenting a rapid method for detection of Bacillus cereus, Listeria monocytogenes and Campylobacter jejuni in food samples

    doi: 10.22038/IJBMS.2017.9275

    Figure Lengend Snippet: Study of the sensitivity of PCR on 1%5 polyacrylamide gel electrophoresis Lane 7: 100 bp DNA marker Lane 1-6: PCR products with the different dilutions of L. monocytogenes, B. cereus and C . jejuni with 10 1 -10 6

    Article Snippet: L. monocytogenes bacterium obtained from Iranian Centre of Industrial and Medical Fungi and Bacteria Collection (PTCC 1298) Following bacteria were prepared from the Iranian Reference Laboratory: Salmonella typhi ATCC 700931, Shigella dysentery ATCC 13313, Yersinia pestis ATCCBAA-1511D-5, Staphylococcus aureus ATCC 25923 , Clostridium perfringes ATCC 13124, Clostridium botulinum ATCC 3502, Vibrio cholera ATCC 9459.

    Techniques: Polymerase Chain Reaction, Polyacrylamide Gel Electrophoresis, Marker

    Lit2 is a functional lipoprotein intramolecular transacylase. (A) The linked lnt ::Spt r and chiQ ::Apr r markers were cotransduced by P1 vir into recipient strains carrying pUC19, p lnt , p Eflit2 , or p Lmlit2 . The percentage of transductants resistant to apramycin alone versus both apramycin and spectinomycin are shown ( n = 24 to 40) and compared to negative (pUC19) and positive (p lnt ). (B) Trypsinized N-terminal lipopeptides of Lpp from the lnt -null strain KA811 expressing E. faecalis lit2 were analyzed by MALDI-TOF MS. The protonated m/z 1,157.8 and sodiated m/z 1,179.8 parent ions are shown (inset), with the latter further fragmented by MS-MS. (C) This spectrum was used to assign Lpp structure from KA811 as the lyso form. Note that the m/z 1,185.8 peak in the parent spectrum is consistent with a C 16:0 , C 18:1 acyl chain combination. (D and F) Trypsinized N-terminal lipopeptides of the lipoprotein KO07_11695 from wild-type L. monocytogenes ATCC 19115 (D) and the corresponding L. monocytogenes lit2 -expressing strain KA849 (F) were analyzed by MALDI-TOF MS. The parent spectra (insets) display the protonated m/z 1,260.9 and sodiated m/z 1,282.9 parent ions. (E and G) The sodiated ions were fragmented by MS-MS (D and F), revealing production of DA-LP in KA847 (E) and the lyso-LP when L. monocytogenes lit2 was expressed (G). (D and F) The structurally diagnostic dehydroalanyl ions for the diacylglycerol-modified (triangle) and the lyso (circle) lipopeptides are indicated.

    Journal: Journal of Bacteriology

    Article Title: Copper-Induced Expression of a Transmissible Lipoprotein Intramolecular Transacylase Alters Lipoprotein Acylation and the Toll-Like Receptor 2 Response to Listeria monocytogenes

    doi: 10.1128/JB.00195-19

    Figure Lengend Snippet: Lit2 is a functional lipoprotein intramolecular transacylase. (A) The linked lnt ::Spt r and chiQ ::Apr r markers were cotransduced by P1 vir into recipient strains carrying pUC19, p lnt , p Eflit2 , or p Lmlit2 . The percentage of transductants resistant to apramycin alone versus both apramycin and spectinomycin are shown ( n = 24 to 40) and compared to negative (pUC19) and positive (p lnt ). (B) Trypsinized N-terminal lipopeptides of Lpp from the lnt -null strain KA811 expressing E. faecalis lit2 were analyzed by MALDI-TOF MS. The protonated m/z 1,157.8 and sodiated m/z 1,179.8 parent ions are shown (inset), with the latter further fragmented by MS-MS. (C) This spectrum was used to assign Lpp structure from KA811 as the lyso form. Note that the m/z 1,185.8 peak in the parent spectrum is consistent with a C 16:0 , C 18:1 acyl chain combination. (D and F) Trypsinized N-terminal lipopeptides of the lipoprotein KO07_11695 from wild-type L. monocytogenes ATCC 19115 (D) and the corresponding L. monocytogenes lit2 -expressing strain KA849 (F) were analyzed by MALDI-TOF MS. The parent spectra (insets) display the protonated m/z 1,260.9 and sodiated m/z 1,282.9 parent ions. (E and G) The sodiated ions were fragmented by MS-MS (D and F), revealing production of DA-LP in KA847 (E) and the lyso-LP when L. monocytogenes lit2 was expressed (G). (D and F) The structurally diagnostic dehydroalanyl ions for the diacylglycerol-modified (triangle) and the lyso (circle) lipopeptides are indicated.

    Article Snippet: To show that L. monocytogenes CFSAN023459 Lit2 similarly functions as an intramolecular transacylase, L. monocytogenes lit2 was cloned under a xylose-inducible promoter into L. monocytogenes ATCC 19115, a type strain of L. monocytogenes that lacks any lit sequence orthologs.

    Techniques: Functional Assay, Single-particle Tracking, Expressing, Mass Spectrometry, Diagnostic Assay, Modification

    Copper induces expression of the lit2 -copper resistance operon. (A and B) RNA was extracted from E. faecalis TX1342 cells grown with 0, 0.1, or 1.0 mM CuCl 2 (A) and L. monocytogenes CFSAN023459 cells grown with 0 or 1.0 mM CuCl 2 (B) and probed for expression of the indicated genes. Chromosomal genes are indicated by asterisks. (C) RNA was extracted from E. faecalis TX1342 (top) and L. monocytogenes CFSAN023459 (bottom) cells induced with various metal ions and probed for lit2 expression. Estimated transcript lengths are indicated. (D and E) Expression of the indicated target genes was measured by RT-qPCR from E. faecalis TX1342 (D) and L. monocytogenes CFSAN023459 (E) cells grown with 0, 0.1, or 1.0 mM copper. Expression was normalized in each strain against the internal control gene gyrA . The data are shown as the means ± standard deviations of the results of three replicates.

    Journal: Journal of Bacteriology

    Article Title: Copper-Induced Expression of a Transmissible Lipoprotein Intramolecular Transacylase Alters Lipoprotein Acylation and the Toll-Like Receptor 2 Response to Listeria monocytogenes

    doi: 10.1128/JB.00195-19

    Figure Lengend Snippet: Copper induces expression of the lit2 -copper resistance operon. (A and B) RNA was extracted from E. faecalis TX1342 cells grown with 0, 0.1, or 1.0 mM CuCl 2 (A) and L. monocytogenes CFSAN023459 cells grown with 0 or 1.0 mM CuCl 2 (B) and probed for expression of the indicated genes. Chromosomal genes are indicated by asterisks. (C) RNA was extracted from E. faecalis TX1342 (top) and L. monocytogenes CFSAN023459 (bottom) cells induced with various metal ions and probed for lit2 expression. Estimated transcript lengths are indicated. (D and E) Expression of the indicated target genes was measured by RT-qPCR from E. faecalis TX1342 (D) and L. monocytogenes CFSAN023459 (E) cells grown with 0, 0.1, or 1.0 mM copper. Expression was normalized in each strain against the internal control gene gyrA . The data are shown as the means ± standard deviations of the results of three replicates.

    Article Snippet: To show that L. monocytogenes CFSAN023459 Lit2 similarly functions as an intramolecular transacylase, L. monocytogenes lit2 was cloned under a xylose-inducible promoter into L. monocytogenes ATCC 19115, a type strain of L. monocytogenes that lacks any lit sequence orthologs.

    Techniques: Expressing, Quantitative RT-PCR

    TLR2 response to whole bacteria. (A) HEK-Blue-TLR2/1/6 cells were exposed to 5-fold dilutions of heat-inactivated whole bacterial cells of L. monocytogenes CFSAN023459 (wild type [WT]) grown with or without 1 mM copper, as well as the derivative Δ lit2 cells grown with or without 1 mM CuCl 2 . (B) HEK-Blue-TLR2/1/6 cells were pretreated with 10 μg/ml of TLR-neutralizing antibodies and then exposed to the same bacterial cell preparations as in panel A. The “WT + 1 mM Cu” sample was added to 8.0 × 10 5 CFU/ml, while the others were added to 3.2 × 10 4 CFU/ml. The percent activity normalized to the water control is indicated. (C and D) HEK-Blue-TLR2/1/6 cells were exposed to 5-fold dilutions of heat-inactivated, whole bacterial cells of strains L. monocytogenes CFSAN023459 (C) and E. faecalis ATCC 19433 (D). The data are shown as the means ± standard deviations of the results of three biological replicates.

    Journal: Journal of Bacteriology

    Article Title: Copper-Induced Expression of a Transmissible Lipoprotein Intramolecular Transacylase Alters Lipoprotein Acylation and the Toll-Like Receptor 2 Response to Listeria monocytogenes

    doi: 10.1128/JB.00195-19

    Figure Lengend Snippet: TLR2 response to whole bacteria. (A) HEK-Blue-TLR2/1/6 cells were exposed to 5-fold dilutions of heat-inactivated whole bacterial cells of L. monocytogenes CFSAN023459 (wild type [WT]) grown with or without 1 mM copper, as well as the derivative Δ lit2 cells grown with or without 1 mM CuCl 2 . (B) HEK-Blue-TLR2/1/6 cells were pretreated with 10 μg/ml of TLR-neutralizing antibodies and then exposed to the same bacterial cell preparations as in panel A. The “WT + 1 mM Cu” sample was added to 8.0 × 10 5 CFU/ml, while the others were added to 3.2 × 10 4 CFU/ml. The percent activity normalized to the water control is indicated. (C and D) HEK-Blue-TLR2/1/6 cells were exposed to 5-fold dilutions of heat-inactivated, whole bacterial cells of strains L. monocytogenes CFSAN023459 (C) and E. faecalis ATCC 19433 (D). The data are shown as the means ± standard deviations of the results of three biological replicates.

    Article Snippet: To show that L. monocytogenes CFSAN023459 Lit2 similarly functions as an intramolecular transacylase, L. monocytogenes lit2 was cloned under a xylose-inducible promoter into L. monocytogenes ATCC 19115, a type strain of L. monocytogenes that lacks any lit sequence orthologs.

    Techniques: Activity Assay

    Copper induces conversion of lipoproteins from DA-LP to lyso-LP. (A and B) MS-MS spectra of the protonated m/z 1,260 (A) and sodiated m/z 1,282 (B) parent ions of lipoprotein KO07_11695 from L. monocytogenes CFSAN023459 grown in the absence of copper. (C and D) MS-MS spectra of the protonated m/z 1,260 (C) and sodiated m/z 1,282 (D) parent ions of L. monocytogenes strain CFSAN023459 grown with 1 mM copper. (E and F) MS-MS spectra of the protonated m/z 1,260 (E) and sodiated m/z 1,282 (F) parent ions of the Δ lit2 strain grown with 1 mM CuCl 2 . (G and H) MS-MS spectra of the protonated m/z 1,260 (G) and sodiated m/z 1,282 (H) parent ions of the Δ lit2 strain back-complemented with pP xyl Lmlit2 grown with 1 mM CuCl 2 and 2% xylose. The diagnostic dehydroalanyl ions for the diacylglycerol-modified (triangles) and the lyso (circles) lipopeptides are indicated. The parent spectra are shown in Fig. S3.

    Journal: Journal of Bacteriology

    Article Title: Copper-Induced Expression of a Transmissible Lipoprotein Intramolecular Transacylase Alters Lipoprotein Acylation and the Toll-Like Receptor 2 Response to Listeria monocytogenes

    doi: 10.1128/JB.00195-19

    Figure Lengend Snippet: Copper induces conversion of lipoproteins from DA-LP to lyso-LP. (A and B) MS-MS spectra of the protonated m/z 1,260 (A) and sodiated m/z 1,282 (B) parent ions of lipoprotein KO07_11695 from L. monocytogenes CFSAN023459 grown in the absence of copper. (C and D) MS-MS spectra of the protonated m/z 1,260 (C) and sodiated m/z 1,282 (D) parent ions of L. monocytogenes strain CFSAN023459 grown with 1 mM copper. (E and F) MS-MS spectra of the protonated m/z 1,260 (E) and sodiated m/z 1,282 (F) parent ions of the Δ lit2 strain grown with 1 mM CuCl 2 . (G and H) MS-MS spectra of the protonated m/z 1,260 (G) and sodiated m/z 1,282 (H) parent ions of the Δ lit2 strain back-complemented with pP xyl Lmlit2 grown with 1 mM CuCl 2 and 2% xylose. The diagnostic dehydroalanyl ions for the diacylglycerol-modified (triangles) and the lyso (circles) lipopeptides are indicated. The parent spectra are shown in Fig. S3.

    Article Snippet: To show that L. monocytogenes CFSAN023459 Lit2 similarly functions as an intramolecular transacylase, L. monocytogenes lit2 was cloned under a xylose-inducible promoter into L. monocytogenes ATCC 19115, a type strain of L. monocytogenes that lacks any lit sequence orthologs.

    Techniques: Mass Spectrometry, Diagnostic Assay, Modification

    ). Other strains were assumed to make the lyso form based on the presence of lit1 . The strains E. faecalis TX1342 and L. monocytogenes CFSAN023459, characterized in this study, are boxed. Plasmid-borne copies of lit2 are indicated with a dagger and chromosomally integrated copies with a triangle. (B) Schematic representations of the E. faecalis TX1342 and L. monocytogenes CFSAN023459 lit2 -copper resistance operons with percent ortholog sequence identities indicated. The proteins with unknown functions are designated “1” and “2” for clarity, and the incomplete CopB2 fragment is designated copB2′ . A predicted rho-independent terminator following copZ is indicated by a vertical line.

    Journal: Journal of Bacteriology

    Article Title: Copper-Induced Expression of a Transmissible Lipoprotein Intramolecular Transacylase Alters Lipoprotein Acylation and the Toll-Like Receptor 2 Response to Listeria monocytogenes

    doi: 10.1128/JB.00195-19

    Figure Lengend Snippet: ). Other strains were assumed to make the lyso form based on the presence of lit1 . The strains E. faecalis TX1342 and L. monocytogenes CFSAN023459, characterized in this study, are boxed. Plasmid-borne copies of lit2 are indicated with a dagger and chromosomally integrated copies with a triangle. (B) Schematic representations of the E. faecalis TX1342 and L. monocytogenes CFSAN023459 lit2 -copper resistance operons with percent ortholog sequence identities indicated. The proteins with unknown functions are designated “1” and “2” for clarity, and the incomplete CopB2 fragment is designated copB2′ . A predicted rho-independent terminator following copZ is indicated by a vertical line.

    Article Snippet: To show that L. monocytogenes CFSAN023459 Lit2 similarly functions as an intramolecular transacylase, L. monocytogenes lit2 was cloned under a xylose-inducible promoter into L. monocytogenes ATCC 19115, a type strain of L. monocytogenes that lacks any lit sequence orthologs.

    Techniques: Plasmid Preparation, Sequencing

    L. monocytogenes induces local degranulation of primary granules. Neutrophils were incubated with L. monocytogenes ( Lm ) for 1 min, washed, and incubated for a total of 3, 5, 10, and 30 min at 37°C in 10% DS/HBSS + . Cells were fixed, and Lm and CD63 were then fluorescently labeled in nonpermeabilized and permeabilized cells to distinguish granule fusion with the plasma membrane from fusion with sealed phagosomes, respectively. ( A ) Representative images of extracellular ( two top panels ) and intracellular ( two bottom panels ) Lm colocalizing with CD63 at 3 and 10 min. ( B ) MFI ± SEM of CD63 on nonpermeabilized neutrophils was measured by quantitative fluorescence microscopy. Data from a representative experiment, of two, are shown.

    Journal: The Journal of Immunology Author Choice

    Article Title: The Pore-Forming Toxin Listeriolysin O Is Degraded by Neutrophil Metalloproteinase-8 and Fails To Mediate Listeria monocytogenes Intracellular Survival in Neutrophils

    doi: 10.4049/jimmunol.1301302

    Figure Lengend Snippet: L. monocytogenes induces local degranulation of primary granules. Neutrophils were incubated with L. monocytogenes ( Lm ) for 1 min, washed, and incubated for a total of 3, 5, 10, and 30 min at 37°C in 10% DS/HBSS + . Cells were fixed, and Lm and CD63 were then fluorescently labeled in nonpermeabilized and permeabilized cells to distinguish granule fusion with the plasma membrane from fusion with sealed phagosomes, respectively. ( A ) Representative images of extracellular ( two top panels ) and intracellular ( two bottom panels ) Lm colocalizing with CD63 at 3 and 10 min. ( B ) MFI ± SEM of CD63 on nonpermeabilized neutrophils was measured by quantitative fluorescence microscopy. Data from a representative experiment, of two, are shown.

    Article Snippet: In some assays, neutrophils were preincubated for 5 min at 37°C ± the protease inhibitor α2 -macroglobulin (300 nM) before adding wt L. monocytogenes at a MOI of 10 in serum-free HBSS+ , then cell culture plates were centrifuged (230 × g ) for 5 min at RT.

    Techniques: Incubation, Labeling, Fluorescence, Microscopy

    Neutrophils kill L. monocytogenes regardless of LLO expression. L. monocytogenes ( Lm ; wt, Δ hly , and LLO + ) were incubated alone ( Lm ) or with neutrophils ( Lm + PMNs) in 10% DS/HBSS + at 37°C at a MOI of 1 ( A ) and 10 ( B , C ). At the indicated times, Triton X-100 was added, and cell lysates were plated to enumerate CFUs. This measured the viability of L. monocytogenes in the absence ( Lm ) and the presence ( Lm + PMNs [Total]) of neutrophils. To measure the viability of intracellular bacteria ( Lm + PMNs [Intra.]), the last hour of incubation at 37°C was carried out with gentamicin. Cells were then washed before lysis and CFU enumeration. (A–C) At least three independent experiments were performed in triplicate. Results are expressed relative to the inoculum. Statistical analyses were performed respective to CFUs enumerated in the absence of neutrophils ( Lm ) at each corresponding time point. * p

    Journal: The Journal of Immunology Author Choice

    Article Title: The Pore-Forming Toxin Listeriolysin O Is Degraded by Neutrophil Metalloproteinase-8 and Fails To Mediate Listeria monocytogenes Intracellular Survival in Neutrophils

    doi: 10.4049/jimmunol.1301302

    Figure Lengend Snippet: Neutrophils kill L. monocytogenes regardless of LLO expression. L. monocytogenes ( Lm ; wt, Δ hly , and LLO + ) were incubated alone ( Lm ) or with neutrophils ( Lm + PMNs) in 10% DS/HBSS + at 37°C at a MOI of 1 ( A ) and 10 ( B , C ). At the indicated times, Triton X-100 was added, and cell lysates were plated to enumerate CFUs. This measured the viability of L. monocytogenes in the absence ( Lm ) and the presence ( Lm + PMNs [Total]) of neutrophils. To measure the viability of intracellular bacteria ( Lm + PMNs [Intra.]), the last hour of incubation at 37°C was carried out with gentamicin. Cells were then washed before lysis and CFU enumeration. (A–C) At least three independent experiments were performed in triplicate. Results are expressed relative to the inoculum. Statistical analyses were performed respective to CFUs enumerated in the absence of neutrophils ( Lm ) at each corresponding time point. * p

    Article Snippet: In some assays, neutrophils were preincubated for 5 min at 37°C ± the protease inhibitor α2 -macroglobulin (300 nM) before adding wt L. monocytogenes at a MOI of 10 in serum-free HBSS+ , then cell culture plates were centrifuged (230 × g ) for 5 min at RT.

    Techniques: Expressing, Incubation, Lysis

    Model. ( 1 ) LLO monomers secreted by L. monocytogenes bind to the neutrophil plasma membrane and oligomerize into prepore complexes. ( 2 ) LLO prepore complexes undergo conformational changes to form pores, which creates an influx of Ca 2+ . ( 3 ) The rise in intracellular Ca 2+ activates granule exocytosis at the forming phagocytic cup. Degranulation also occurs into the phagosome. Neutrophil MMP-8 rapidly degrades extracellular and intraphagosomal LLO. We propose that these actions inhibit LLO-mediated escape, trapping L. monocytogenes in a bactericidal phagosome. ( 4 ) Degranulation is linked to increased endocytosis in neutrophils. Endocytosis of LLO pores may also protect neutrophils from LLO-mediated membrane damage.

    Journal: The Journal of Immunology Author Choice

    Article Title: The Pore-Forming Toxin Listeriolysin O Is Degraded by Neutrophil Metalloproteinase-8 and Fails To Mediate Listeria monocytogenes Intracellular Survival in Neutrophils

    doi: 10.4049/jimmunol.1301302

    Figure Lengend Snippet: Model. ( 1 ) LLO monomers secreted by L. monocytogenes bind to the neutrophil plasma membrane and oligomerize into prepore complexes. ( 2 ) LLO prepore complexes undergo conformational changes to form pores, which creates an influx of Ca 2+ . ( 3 ) The rise in intracellular Ca 2+ activates granule exocytosis at the forming phagocytic cup. Degranulation also occurs into the phagosome. Neutrophil MMP-8 rapidly degrades extracellular and intraphagosomal LLO. We propose that these actions inhibit LLO-mediated escape, trapping L. monocytogenes in a bactericidal phagosome. ( 4 ) Degranulation is linked to increased endocytosis in neutrophils. Endocytosis of LLO pores may also protect neutrophils from LLO-mediated membrane damage.

    Article Snippet: In some assays, neutrophils were preincubated for 5 min at 37°C ± the protease inhibitor α2 -macroglobulin (300 nM) before adding wt L. monocytogenes at a MOI of 10 in serum-free HBSS+ , then cell culture plates were centrifuged (230 × g ) for 5 min at RT.

    Techniques:

    L. monocytogenes induces exocytosis of secondary and primary granules. ( A ) Neutrophils were incubated for 30 min at 37°C with L. monocytogenes or L. innocua at MOI of 10, washed, and labeled on ice with FITC-conjugated anti-CD66b, anti-CD63, or isotype control Abs. Cells were fixed postlabeling. ( B ) Neutrophils were incubated for 15 min at 37°C with LLO (0.5–5 nM) or LLOpL (50 nM), fixed, and labeled with FITC-conjugated anti-CD66b, anti-CD63, or isotype control Abs. (A and B) Neutrophil MFI was measured by flow cytometry. Data are expressed as MFI relative to untreated cells. Data from a representative experiment, of three, are shown.

    Journal: The Journal of Immunology Author Choice

    Article Title: The Pore-Forming Toxin Listeriolysin O Is Degraded by Neutrophil Metalloproteinase-8 and Fails To Mediate Listeria monocytogenes Intracellular Survival in Neutrophils

    doi: 10.4049/jimmunol.1301302

    Figure Lengend Snippet: L. monocytogenes induces exocytosis of secondary and primary granules. ( A ) Neutrophils were incubated for 30 min at 37°C with L. monocytogenes or L. innocua at MOI of 10, washed, and labeled on ice with FITC-conjugated anti-CD66b, anti-CD63, or isotype control Abs. Cells were fixed postlabeling. ( B ) Neutrophils were incubated for 15 min at 37°C with LLO (0.5–5 nM) or LLOpL (50 nM), fixed, and labeled with FITC-conjugated anti-CD66b, anti-CD63, or isotype control Abs. (A and B) Neutrophil MFI was measured by flow cytometry. Data are expressed as MFI relative to untreated cells. Data from a representative experiment, of three, are shown.

    Article Snippet: In some assays, neutrophils were preincubated for 5 min at 37°C ± the protease inhibitor α2 -macroglobulin (300 nM) before adding wt L. monocytogenes at a MOI of 10 in serum-free HBSS+ , then cell culture plates were centrifuged (230 × g ) for 5 min at RT.

    Techniques: Incubation, Labeling, Flow Cytometry, Cytometry

    Rescue of the elektra phenotype by BAC transgenesis Transgenic mice were produced by microinjecting a BAC clone containing the wild-type Slfn2 sequence into single-cell embryos homozygous for the elektra mutation. (a) DNA sequence of the BAC clone and genomic DNA from an elektra homozygote in the region of the Slfn2 elektra mutation. The BAC clone contains the wild-type Slfn2 sequence. (b) Flow cytometric analysis of CD8 and CD4 staining of blood from a WT mouse, a homozygous elektra transgenic mouse carrying the Slfn2 transgene ( elektra-BAC Tg ), and a littermate lacking the transgene ( elektra ). Results are representative of 4 mice per genotype. (c) Survival curves for transgenic mice ( elektra-BAC Tg , n =7), littermates without the transgene ( elektra , n =4), and WT mice ( n =5) upon challenge with 2 × 10 5 PFU of MCMV. (d) Survival curves for transgenic mice ( elektra-BAC Tg , n =7), littermates without the transgene ( elektra , n =5), and WT mice ( n =7) upon challenge with 5 × 10 5 CFU of L. monocytogenes .

    Journal: Nature immunology

    Article Title: A Slfn2 mutation causes lymphoid and myeloid immunodeficiency due to loss of immune cell quiescence

    doi: 10.1038/ni.1847

    Figure Lengend Snippet: Rescue of the elektra phenotype by BAC transgenesis Transgenic mice were produced by microinjecting a BAC clone containing the wild-type Slfn2 sequence into single-cell embryos homozygous for the elektra mutation. (a) DNA sequence of the BAC clone and genomic DNA from an elektra homozygote in the region of the Slfn2 elektra mutation. The BAC clone contains the wild-type Slfn2 sequence. (b) Flow cytometric analysis of CD8 and CD4 staining of blood from a WT mouse, a homozygous elektra transgenic mouse carrying the Slfn2 transgene ( elektra-BAC Tg ), and a littermate lacking the transgene ( elektra ). Results are representative of 4 mice per genotype. (c) Survival curves for transgenic mice ( elektra-BAC Tg , n =7), littermates without the transgene ( elektra , n =4), and WT mice ( n =5) upon challenge with 2 × 10 5 PFU of MCMV. (d) Survival curves for transgenic mice ( elektra-BAC Tg , n =7), littermates without the transgene ( elektra , n =5), and WT mice ( n =7) upon challenge with 5 × 10 5 CFU of L. monocytogenes .

    Article Snippet: For in vivo challenge with L. monocytogenes (strain 10403S; Xenogen, Inc.), bioluminescent bacteria were prepared and injected intravenously as described previously .

    Techniques: BAC Assay, Transgenic Assay, Mouse Assay, Produced, Sequencing, Mutagenesis, Flow Cytometry, Staining

    Apoptosis of homozygous elektra monocytes in response to activation signals (a) Left, Ly6C versus CD11b staining of splenocytes from uninfected or L. monocytogenes infected WT or elektra mice ( n =5 per genotype). Percentages of inflammatory monocytes and neutrophils are indicated by R1 and R2 gated cell populations, respectively. Results are representative of 3 experiments. Right, average percentage of inflammatory monocytes 48 h post-infection from spleen, blood, and bone marrow of WT or elektra mice ( n =5 each). (b) Flow cytometric analysis of WT (CD45.1 + ) and elektra (CD45.2 + ) donor CD4 + , CD8 + , B cells (B220 + ), and inflammatory monocytes (gated on CD11b + population) in the blood of CD3-deficient mixed bone marrow chimeras ( n =3). Results are representative of two experiments. (c–e) Isolated bone marrow inflammatory monocytes from WT or elektra mice ( n =2 each) cultured for 3 days in medium (untreated) or in medium supplemented with IFN-γ and heat-killed L. monocytogenes (treated). (c) Top, MHC class II staining. Bottom, nitric oxide concentration. (d) Forward-scatter (FSC) versus side-scatter (SSC) profile. Populations of live and dead cells are indicated. (e) Annexin V staining of the gated high FSC population. Results are representative of three experiments. For all panels, *** P

    Journal: Nature immunology

    Article Title: A Slfn2 mutation causes lymphoid and myeloid immunodeficiency due to loss of immune cell quiescence

    doi: 10.1038/ni.1847

    Figure Lengend Snippet: Apoptosis of homozygous elektra monocytes in response to activation signals (a) Left, Ly6C versus CD11b staining of splenocytes from uninfected or L. monocytogenes infected WT or elektra mice ( n =5 per genotype). Percentages of inflammatory monocytes and neutrophils are indicated by R1 and R2 gated cell populations, respectively. Results are representative of 3 experiments. Right, average percentage of inflammatory monocytes 48 h post-infection from spleen, blood, and bone marrow of WT or elektra mice ( n =5 each). (b) Flow cytometric analysis of WT (CD45.1 + ) and elektra (CD45.2 + ) donor CD4 + , CD8 + , B cells (B220 + ), and inflammatory monocytes (gated on CD11b + population) in the blood of CD3-deficient mixed bone marrow chimeras ( n =3). Results are representative of two experiments. (c–e) Isolated bone marrow inflammatory monocytes from WT or elektra mice ( n =2 each) cultured for 3 days in medium (untreated) or in medium supplemented with IFN-γ and heat-killed L. monocytogenes (treated). (c) Top, MHC class II staining. Bottom, nitric oxide concentration. (d) Forward-scatter (FSC) versus side-scatter (SSC) profile. Populations of live and dead cells are indicated. (e) Annexin V staining of the gated high FSC population. Results are representative of three experiments. For all panels, *** P

    Article Snippet: For in vivo challenge with L. monocytogenes (strain 10403S; Xenogen, Inc.), bioluminescent bacteria were prepared and injected intravenously as described previously .

    Techniques: Activation Assay, Staining, Infection, Mouse Assay, Flow Cytometry, Isolation, Cell Culture, Concentration Assay

    Homozygous elektra mutants are highly susceptible to MCMV, LCMV and L. monocytogenes infections (a) Survival curves for WT ( n =8) and homozygous elektra mutants ( n =8) upon challenge with 2 × 10 5 PFU of MCMV. Results are representative of five independent experiments. (b) Survival curves upon infection with 2 × 10 5 PFU of MCMV after reciprocal bone marrow transplantation. Recipient mice were reconstituted with 5 × 10 6 bone marrow cells 1 day after 10-Gy dose of irradiation. Congenic C57BL/6.SJL ( Ptprc a Pep3 b ; Ly5.1 + ), C57BL/6J ( Ptprc b Pep3 a ; Ly5.2 + ) WT or elektra mutant mice were used as both recipients and donors as indicated in the figure. C57BL/6J into C57BL/6J, n =3; C57BL/6J into elektra , n =6; elektra into C57BL/6J, n =6; elektra into elektra , n =3. Results are representative of 2 independent experiments. (c) WT and homozygous elektra mice (3 mice in each group) were i.v. injected with either 200 or 2 × 10 6 PFU of LCMV (Armstrong strain). Viral load was measured in spleens 7 days after injection. Results are representative of two independent experiments. ** P

    Journal: Nature immunology

    Article Title: A Slfn2 mutation causes lymphoid and myeloid immunodeficiency due to loss of immune cell quiescence

    doi: 10.1038/ni.1847

    Figure Lengend Snippet: Homozygous elektra mutants are highly susceptible to MCMV, LCMV and L. monocytogenes infections (a) Survival curves for WT ( n =8) and homozygous elektra mutants ( n =8) upon challenge with 2 × 10 5 PFU of MCMV. Results are representative of five independent experiments. (b) Survival curves upon infection with 2 × 10 5 PFU of MCMV after reciprocal bone marrow transplantation. Recipient mice were reconstituted with 5 × 10 6 bone marrow cells 1 day after 10-Gy dose of irradiation. Congenic C57BL/6.SJL ( Ptprc a Pep3 b ; Ly5.1 + ), C57BL/6J ( Ptprc b Pep3 a ; Ly5.2 + ) WT or elektra mutant mice were used as both recipients and donors as indicated in the figure. C57BL/6J into C57BL/6J, n =3; C57BL/6J into elektra , n =6; elektra into C57BL/6J, n =6; elektra into elektra , n =3. Results are representative of 2 independent experiments. (c) WT and homozygous elektra mice (3 mice in each group) were i.v. injected with either 200 or 2 × 10 6 PFU of LCMV (Armstrong strain). Viral load was measured in spleens 7 days after injection. Results are representative of two independent experiments. ** P

    Article Snippet: For in vivo challenge with L. monocytogenes (strain 10403S; Xenogen, Inc.), bioluminescent bacteria were prepared and injected intravenously as described previously .

    Techniques: Infection, Transplantation Assay, Mouse Assay, Irradiation, Mutagenesis, Injection

    ApaI/AscI PFGE profiles of L. monocytogenes strains 4b and IVb-v1. The dendrogram was calculated and drawn using Bionumerics software. NSW; New South Wales, VIC; Victoria, CA; California, CT; Connecticut, and NK;Not known.

    Journal: PLoS ONE

    Article Title: Genomic Characterization of Novel Listeria monocytogenes Serotype 4b Variant Strains

    doi: 10.1371/journal.pone.0089024

    Figure Lengend Snippet: ApaI/AscI PFGE profiles of L. monocytogenes strains 4b and IVb-v1. The dendrogram was calculated and drawn using Bionumerics software. NSW; New South Wales, VIC; Victoria, CA; California, CT; Connecticut, and NK;Not known.

    Article Snippet: Dispersion of unique genotypes of L. monocytogenes throughout the world has been illustrated by the occurrence of multiple epidemic clones (EC) and multiple sequence types (ST) , .

    Techniques: Software

    Hierarchical clustering based on the Robust Multi Array (RMA) analysis of the L. monocytogenes strains serotypes 4b (LS406, LS412) and IVb-v1 (LS542, LS642, LS643, LS644, LS645).

    Journal: PLoS ONE

    Article Title: Genomic Characterization of Novel Listeria monocytogenes Serotype 4b Variant Strains

    doi: 10.1371/journal.pone.0089024

    Figure Lengend Snippet: Hierarchical clustering based on the Robust Multi Array (RMA) analysis of the L. monocytogenes strains serotypes 4b (LS406, LS412) and IVb-v1 (LS542, LS642, LS643, LS644, LS645).

    Article Snippet: Dispersion of unique genotypes of L. monocytogenes throughout the world has been illustrated by the occurrence of multiple epidemic clones (EC) and multiple sequence types (ST) , .

    Techniques:

    Multiplex PCR profiles of L. monocytogenes strains. Lanes: MW, Molecular weight markers. Lane 1: LS1 serotype 1/2a, Lane 2: LS402, serotype 4b; Lane3-7: serotype IVb-v1 strains LS542; LS642; LS643; LS644; LS645, respectively.

    Journal: PLoS ONE

    Article Title: Genomic Characterization of Novel Listeria monocytogenes Serotype 4b Variant Strains

    doi: 10.1371/journal.pone.0089024

    Figure Lengend Snippet: Multiplex PCR profiles of L. monocytogenes strains. Lanes: MW, Molecular weight markers. Lane 1: LS1 serotype 1/2a, Lane 2: LS402, serotype 4b; Lane3-7: serotype IVb-v1 strains LS542; LS642; LS643; LS644; LS645, respectively.

    Article Snippet: Dispersion of unique genotypes of L. monocytogenes throughout the world has been illustrated by the occurrence of multiple epidemic clones (EC) and multiple sequence types (ST) , .

    Techniques: Multiplex Assay, Polymerase Chain Reaction, Molecular Weight

    The organization of the genomic region containing the lmo 0737 to lmo0739 cassette that is present only in the L. monocytogenes serotypes IVb-v1 and 1/2a strains, confirmed by microarray analysis.

    Journal: PLoS ONE

    Article Title: Genomic Characterization of Novel Listeria monocytogenes Serotype 4b Variant Strains

    doi: 10.1371/journal.pone.0089024

    Figure Lengend Snippet: The organization of the genomic region containing the lmo 0737 to lmo0739 cassette that is present only in the L. monocytogenes serotypes IVb-v1 and 1/2a strains, confirmed by microarray analysis.

    Article Snippet: Dispersion of unique genotypes of L. monocytogenes throughout the world has been illustrated by the occurrence of multiple epidemic clones (EC) and multiple sequence types (ST) , .

    Techniques: Microarray

    Temporal distribution of the 43 ST38 L. monocytogenes clinical isolates collected in Lombardy (2006–2014) according to the two different Virulence Types (VT80 and VT104)

    Journal: BMC Infectious Diseases

    Article Title: Identification of a major Listeria monocytogenes outbreak clone linked to soft cheese in Northern Italy – 2009-2011

    doi: 10.1186/s12879-017-2441-6

    Figure Lengend Snippet: Temporal distribution of the 43 ST38 L. monocytogenes clinical isolates collected in Lombardy (2006–2014) according to the two different Virulence Types (VT80 and VT104)

    Article Snippet: The specific serotype within the serogroups was then determined by conventional serotyping, using L. monocytogenes antisera (Denka Seiken Co., Ltd. Tokio, Japan), according to the manufacturer’s instructions.

    Techniques:

    Pulsed field gel electrophoresis profiles of the L. monocytogenes isolates identified as ST38 with official Asc I- Apa I pulsotype codes as provided by The European Surveillance System

    Journal: BMC Infectious Diseases

    Article Title: Identification of a major Listeria monocytogenes outbreak clone linked to soft cheese in Northern Italy – 2009-2011

    doi: 10.1186/s12879-017-2441-6

    Figure Lengend Snippet: Pulsed field gel electrophoresis profiles of the L. monocytogenes isolates identified as ST38 with official Asc I- Apa I pulsotype codes as provided by The European Surveillance System

    Article Snippet: The specific serotype within the serogroups was then determined by conventional serotyping, using L. monocytogenes antisera (Denka Seiken Co., Ltd. Tokio, Japan), according to the manufacturer’s instructions.

    Techniques: Pulsed-Field Gel, Electrophoresis

    Effect of L. monocytogenes LipA on virulence in mice. (A) Groups of 4 to 5 C57BL/6 wt mice were infected intraperitoneally with 2 × 10 6 L. monocytogenes LO28 wt and Δ lipA bacteria. At days 1 and 3, the spleen and liver were homogenized

    Journal: Infection and Immunity

    Article Title: LipA, a Tyrosine and Lipid Phosphatase Involved in the Virulence of Listeria monocytogenes ▿ ▿ †

    doi: 10.1128/IAI.05073-11

    Figure Lengend Snippet: Effect of L. monocytogenes LipA on virulence in mice. (A) Groups of 4 to 5 C57BL/6 wt mice were infected intraperitoneally with 2 × 10 6 L. monocytogenes LO28 wt and Δ lipA bacteria. At days 1 and 3, the spleen and liver were homogenized

    Article Snippet: L. monocytogenes was transformed with plasmid pAM401(LipA-GFP) or pAM401(pAM40-GFP).

    Techniques: Mouse Assay, Infection

    Expression of L. monocytogenes lipA in human blood. Transcriptional tiling maps of L. monocytogenes EGDe grown in BHI to exponential phase at 37°C (black dots) or in human blood at 37°C for 60 min (red dots) show normalized hybridization

    Journal: Infection and Immunity

    Article Title: LipA, a Tyrosine and Lipid Phosphatase Involved in the Virulence of Listeria monocytogenes ▿ ▿ †

    doi: 10.1128/IAI.05073-11

    Figure Lengend Snippet: Expression of L. monocytogenes lipA in human blood. Transcriptional tiling maps of L. monocytogenes EGDe grown in BHI to exponential phase at 37°C (black dots) or in human blood at 37°C for 60 min (red dots) show normalized hybridization

    Article Snippet: L. monocytogenes was transformed with plasmid pAM401(LipA-GFP) or pAM401(pAM40-GFP).

    Techniques: Expressing, Hybridization

    LipA secretion and phosphatase activity of recombinant LipA. (A) The supernatant, membrane, and cytosolic fraction were collected from overnight cultures of L. monocytogenes LO28 clones, containing an empty plasmid (C), a plasmid expressing only GFP (GFP),

    Journal: Infection and Immunity

    Article Title: LipA, a Tyrosine and Lipid Phosphatase Involved in the Virulence of Listeria monocytogenes ▿ ▿ †

    doi: 10.1128/IAI.05073-11

    Figure Lengend Snippet: LipA secretion and phosphatase activity of recombinant LipA. (A) The supernatant, membrane, and cytosolic fraction were collected from overnight cultures of L. monocytogenes LO28 clones, containing an empty plasmid (C), a plasmid expressing only GFP (GFP),

    Article Snippet: L. monocytogenes was transformed with plasmid pAM401(LipA-GFP) or pAM401(pAM40-GFP).

    Techniques: Activity Assay, Recombinant, Clone Assay, Plasmid Preparation, Expressing

    L. monocytogenes LipA. (A) Genomic organization of the lipA gene region. Data from the complete genome sequence of L. monocytogenes EGDe were used to draw the map. Hairpins depict putative terminators. The ffh gene encodes the signal recognition particle,

    Journal: Infection and Immunity

    Article Title: LipA, a Tyrosine and Lipid Phosphatase Involved in the Virulence of Listeria monocytogenes ▿ ▿ †

    doi: 10.1128/IAI.05073-11

    Figure Lengend Snippet: L. monocytogenes LipA. (A) Genomic organization of the lipA gene region. Data from the complete genome sequence of L. monocytogenes EGDe were used to draw the map. Hairpins depict putative terminators. The ffh gene encodes the signal recognition particle,

    Article Snippet: L. monocytogenes was transformed with plasmid pAM401(LipA-GFP) or pAM401(pAM40-GFP).

    Techniques: Sequencing

    Effect of L. monocytogenes LipA on intracellular growth. Growth of L. monocytogenes LO28 or EGDe was assessed by plating serial dilutions of cellular lysates at the indicated time points. Numbers of CFU are displayed as log CFU per 1 × 10 6 cells.

    Journal: Infection and Immunity

    Article Title: LipA, a Tyrosine and Lipid Phosphatase Involved in the Virulence of Listeria monocytogenes ▿ ▿ †

    doi: 10.1128/IAI.05073-11

    Figure Lengend Snippet: Effect of L. monocytogenes LipA on intracellular growth. Growth of L. monocytogenes LO28 or EGDe was assessed by plating serial dilutions of cellular lysates at the indicated time points. Numbers of CFU are displayed as log CFU per 1 × 10 6 cells.

    Article Snippet: L. monocytogenes was transformed with plasmid pAM401(LipA-GFP) or pAM401(pAM40-GFP).

    Techniques:

    Effect of L. monocytogenes LipA on movement and cell-to-cell spread. (A) Detection of L. monocytogenes cell-to-cell spread by a plaque assay. L2 cells were infected with L. monocytogenes LO28 wt and Δ lipA strains at an MOI of 0.5. Three days after

    Journal: Infection and Immunity

    Article Title: LipA, a Tyrosine and Lipid Phosphatase Involved in the Virulence of Listeria monocytogenes ▿ ▿ †

    doi: 10.1128/IAI.05073-11

    Figure Lengend Snippet: Effect of L. monocytogenes LipA on movement and cell-to-cell spread. (A) Detection of L. monocytogenes cell-to-cell spread by a plaque assay. L2 cells were infected with L. monocytogenes LO28 wt and Δ lipA strains at an MOI of 0.5. Three days after

    Article Snippet: L. monocytogenes was transformed with plasmid pAM401(LipA-GFP) or pAM401(pAM40-GFP).

    Techniques: Plaque Assay, Infection

    Effect of L. monocytogenes LipA on serum cytokine levels. Levels of IFN-γ, TNF-α, and IL-6 in sera of C57BL/6 wt mice after intraperitoneal infection with L. monocytogenes LO28 wt and Δ lipA strains was measured by ELISA. Cytokine

    Journal: Infection and Immunity

    Article Title: LipA, a Tyrosine and Lipid Phosphatase Involved in the Virulence of Listeria monocytogenes ▿ ▿ †

    doi: 10.1128/IAI.05073-11

    Figure Lengend Snippet: Effect of L. monocytogenes LipA on serum cytokine levels. Levels of IFN-γ, TNF-α, and IL-6 in sera of C57BL/6 wt mice after intraperitoneal infection with L. monocytogenes LO28 wt and Δ lipA strains was measured by ELISA. Cytokine

    Article Snippet: L. monocytogenes was transformed with plasmid pAM401(LipA-GFP) or pAM401(pAM40-GFP).

    Techniques: Mouse Assay, Infection, Enzyme-linked Immunosorbent Assay

    Combined PFGE-cluster analysis of L. monocytogenes isolated from serosa and silage samples

    Journal: BMC Veterinary Research

    Article Title: Listeriosis in fattening pigs caused by poor quality silage - a case report

    doi: 10.1186/s12917-018-1687-6

    Figure Lengend Snippet: Combined PFGE-cluster analysis of L. monocytogenes isolated from serosa and silage samples

    Article Snippet: As just the fibre-reinforced plastic tank was not additionally sealed with plastic, the continuous source of fresh air around the entrance doors may have caused higher pH values due to insufficient ensilaging and therefore may have enhanced moulding and growth of L. monocytogenes .

    Techniques: Isolation

    Distribution of Listeria monocytogenes antigen (brown signals) in the lamina propria mucosae of the colon, pig 1 ( L. monocytogenes immunohistochemistry)

    Journal: BMC Veterinary Research

    Article Title: Listeriosis in fattening pigs caused by poor quality silage - a case report

    doi: 10.1186/s12917-018-1687-6

    Figure Lengend Snippet: Distribution of Listeria monocytogenes antigen (brown signals) in the lamina propria mucosae of the colon, pig 1 ( L. monocytogenes immunohistochemistry)

    Article Snippet: As just the fibre-reinforced plastic tank was not additionally sealed with plastic, the continuous source of fresh air around the entrance doors may have caused higher pH values due to insufficient ensilaging and therefore may have enhanced moulding and growth of L. monocytogenes .

    Techniques: Immunohistochemistry

    ), which can be integrated in a single step into the chromosome of L. monocytogenes . The plasmids were continually selected in appropriate auxotrophic E. coli Δ glnA and L. monocytogenes Δ dal Δ dat strains. pPL2dal retains an antibiotic resistance marker for chloramphenicol resistance ( cat ) in gram-negative bacteria, while pPL2dalGlnA lacks all antibiotic resistance markers.

    Journal: Clinical and Vaccine Immunology : CVI

    Article Title: Stable Integration Vector for Nutrient Broth-Based Selection of Attenuated Listeria monocytogenes Strains with Recombinant Antigen Expression ▿

    doi: 10.1128/CVI.00208-08

    Figure Lengend Snippet: ), which can be integrated in a single step into the chromosome of L. monocytogenes . The plasmids were continually selected in appropriate auxotrophic E. coli Δ glnA and L. monocytogenes Δ dal Δ dat strains. pPL2dal retains an antibiotic resistance marker for chloramphenicol resistance ( cat ) in gram-negative bacteria, while pPL2dalGlnA lacks all antibiotic resistance markers.

    Article Snippet: Brain heart infusion (BHI) broth was used for L. monocytogenes cultures and was purchased from Difco Laboratories (a subsidiary of BD Biosciences, San Jose, CA).

    Techniques: Marker

    T-cell response to exogenous antigens expressed from pPL2dalGlnA in a highly attenuated L. monocytogenes strain. (A) Diagram of the SIV Nef and HIV Tat antigen expression cassette cloned into pPL2dalGlnA and integrated in L. monocytogenes Δ dal Δ dat Δ actA - plcB (LmddΔAB). (B) Western blots showing FLAG-tagged Nef and VSV epitope-tagged Tat proteins in supernatants of the recombinant L. monocytogenes Δ dal Δ dat Δ actA-plcB Nef Tat strain. Shown are immunoblots of proteins precipitated with TCA from the supernatant of L. monocytogenes Δ dal Δ dat Δ actA-plcB Nef Tat strain (lanes 1 and 3) or a control L. monocytogenes Δ dal Δ dat strain (lanes 2 and 4) probed with anti-FLAG tag or anti-VSV tag antibodies. (C) Frequency of IFN-γ-secreting cells responding to overlapping HIV Tat peptides. Shown are the mean number of responding IFN-γ-positive spleoncytes detected by the ELISPOT assay 10 days after the immunization of mice with L. monocytogenes Δ dal Δ dat Δ actA-plcB Nef Tat. The results of the experiments shown are representative of those of at least two additional studies.

    Journal: Clinical and Vaccine Immunology : CVI

    Article Title: Stable Integration Vector for Nutrient Broth-Based Selection of Attenuated Listeria monocytogenes Strains with Recombinant Antigen Expression ▿

    doi: 10.1128/CVI.00208-08

    Figure Lengend Snippet: T-cell response to exogenous antigens expressed from pPL2dalGlnA in a highly attenuated L. monocytogenes strain. (A) Diagram of the SIV Nef and HIV Tat antigen expression cassette cloned into pPL2dalGlnA and integrated in L. monocytogenes Δ dal Δ dat Δ actA - plcB (LmddΔAB). (B) Western blots showing FLAG-tagged Nef and VSV epitope-tagged Tat proteins in supernatants of the recombinant L. monocytogenes Δ dal Δ dat Δ actA-plcB Nef Tat strain. Shown are immunoblots of proteins precipitated with TCA from the supernatant of L. monocytogenes Δ dal Δ dat Δ actA-plcB Nef Tat strain (lanes 1 and 3) or a control L. monocytogenes Δ dal Δ dat strain (lanes 2 and 4) probed with anti-FLAG tag or anti-VSV tag antibodies. (C) Frequency of IFN-γ-secreting cells responding to overlapping HIV Tat peptides. Shown are the mean number of responding IFN-γ-positive spleoncytes detected by the ELISPOT assay 10 days after the immunization of mice with L. monocytogenes Δ dal Δ dat Δ actA-plcB Nef Tat. The results of the experiments shown are representative of those of at least two additional studies.

    Article Snippet: Brain heart infusion (BHI) broth was used for L. monocytogenes cultures and was purchased from Difco Laboratories (a subsidiary of BD Biosciences, San Jose, CA).

    Techniques: Expressing, Clone Assay, Western Blot, Recombinant, FLAG-tag, Enzyme-linked Immunospot, Mouse Assay

    PLS-DA scatter plots of L. monocytogenes inoculated in enriched selective media containing beef sample. (A) PLS-DA Score scatter plot ( R 2 X = 91.5%, R 2 Y = 100%, Q 2 = 91.7%). Green circles (•) indicate data obtained from control beef samples and red stars (★) indicate data from spiked beef samples after 24 h of incubation for L. monocytogenes . The PLS-DA ellipse (solid line) represents the 95% confidence interval. (B) PLS-DA Loading scatter plot.

    Journal: Frontiers in Microbiology

    Article Title: Detection of Foodborne Pathogens Using Proteomics and Metabolomics-Based Approaches

    doi: 10.3389/fmicb.2018.03132

    Figure Lengend Snippet: PLS-DA scatter plots of L. monocytogenes inoculated in enriched selective media containing beef sample. (A) PLS-DA Score scatter plot ( R 2 X = 91.5%, R 2 Y = 100%, Q 2 = 91.7%). Green circles (•) indicate data obtained from control beef samples and red stars (★) indicate data from spiked beef samples after 24 h of incubation for L. monocytogenes . The PLS-DA ellipse (solid line) represents the 95% confidence interval. (B) PLS-DA Loading scatter plot.

    Article Snippet: E. coli (ATCC 8739) was used as a reference strain for external calibration of MALDI-ToF MS. Media included brain heart infusion agar (BHIA, Sigma-Aldrich, Australia), Oxoid novel enrichment (ONE) broth for L. monocytogenes (OBL, Oxoid, Hampshire, United Kingdom), ONE broth for Salmonella (OBS, Oxoid, Hampshire, United Kingdom), Rappaport-Vassiliadis Soya Peptone broth (RVSP, Sigma-Aldrich, NSW, Australia), Muller-Kauffmann tetrathionate broth supplemented with novobiocin (MKTTn, Micromedia, VIC, Australia), E. coli medium (EC, Sigma-Aldrich, NSW, Australia), EC supplement with 4-methylumbelliferyl-β-D-glucuronide (EC-Mug, Sigma-Aldrich, NSW, Australia) and tryptic soy broth supplemented with novobiocin (TSBn, Sigma-Aldrich, NSW, Australia).

    Techniques: Incubation

    Volcano plots of (A) L. monocytogenes , (B) S. enterica and (C) E. coli O157:H7 enriched in selective media containing the minced beef samples. Green circles (•) and red stars (★) indicate statistically significant putative biomarker metabolites found in control beef samples and spiked meat samples, respectively, after specified period of incubation (For L. monocytogenes : 24 h; for S. enterica : 18 h and E. coli O157:H7: 12 h). A few statistically significant metabolites in spiked samples are labeled in the volcano plots (see Tables 2 – 4 for top ten statistically significant metabolites). Please refer to Supplementary Information for list of other statistically significant metabolites. Blue open circles (∘) indicate metabolites that are not statistically significant for the discrimination (see Supplementary Information ). The dashed lines on the volcano plot represent a p -value of 0.05 ( Y -axis) and a fold change of 2 ( X -axis).

    Journal: Frontiers in Microbiology

    Article Title: Detection of Foodborne Pathogens Using Proteomics and Metabolomics-Based Approaches

    doi: 10.3389/fmicb.2018.03132

    Figure Lengend Snippet: Volcano plots of (A) L. monocytogenes , (B) S. enterica and (C) E. coli O157:H7 enriched in selective media containing the minced beef samples. Green circles (•) and red stars (★) indicate statistically significant putative biomarker metabolites found in control beef samples and spiked meat samples, respectively, after specified period of incubation (For L. monocytogenes : 24 h; for S. enterica : 18 h and E. coli O157:H7: 12 h). A few statistically significant metabolites in spiked samples are labeled in the volcano plots (see Tables 2 – 4 for top ten statistically significant metabolites). Please refer to Supplementary Information for list of other statistically significant metabolites. Blue open circles (∘) indicate metabolites that are not statistically significant for the discrimination (see Supplementary Information ). The dashed lines on the volcano plot represent a p -value of 0.05 ( Y -axis) and a fold change of 2 ( X -axis).

    Article Snippet: E. coli (ATCC 8739) was used as a reference strain for external calibration of MALDI-ToF MS. Media included brain heart infusion agar (BHIA, Sigma-Aldrich, Australia), Oxoid novel enrichment (ONE) broth for L. monocytogenes (OBL, Oxoid, Hampshire, United Kingdom), ONE broth for Salmonella (OBS, Oxoid, Hampshire, United Kingdom), Rappaport-Vassiliadis Soya Peptone broth (RVSP, Sigma-Aldrich, NSW, Australia), Muller-Kauffmann tetrathionate broth supplemented with novobiocin (MKTTn, Micromedia, VIC, Australia), E. coli medium (EC, Sigma-Aldrich, NSW, Australia), EC supplement with 4-methylumbelliferyl-β-D-glucuronide (EC-Mug, Sigma-Aldrich, NSW, Australia) and tryptic soy broth supplemented with novobiocin (TSBn, Sigma-Aldrich, NSW, Australia).

    Techniques: Biomarker Assay, Incubation, Labeling

    Characterization of total “identified” metabolites. A total of 104 metabolites for Listeria monocytogenes , 92 metabolites for S. enterica and 104 metabolites for E. coli O157:H7 were identified (see Supplementary Information for list of all identified metabolites).

    Journal: Frontiers in Microbiology

    Article Title: Detection of Foodborne Pathogens Using Proteomics and Metabolomics-Based Approaches

    doi: 10.3389/fmicb.2018.03132

    Figure Lengend Snippet: Characterization of total “identified” metabolites. A total of 104 metabolites for Listeria monocytogenes , 92 metabolites for S. enterica and 104 metabolites for E. coli O157:H7 were identified (see Supplementary Information for list of all identified metabolites).

    Article Snippet: E. coli (ATCC 8739) was used as a reference strain for external calibration of MALDI-ToF MS. Media included brain heart infusion agar (BHIA, Sigma-Aldrich, Australia), Oxoid novel enrichment (ONE) broth for L. monocytogenes (OBL, Oxoid, Hampshire, United Kingdom), ONE broth for Salmonella (OBS, Oxoid, Hampshire, United Kingdom), Rappaport-Vassiliadis Soya Peptone broth (RVSP, Sigma-Aldrich, NSW, Australia), Muller-Kauffmann tetrathionate broth supplemented with novobiocin (MKTTn, Micromedia, VIC, Australia), E. coli medium (EC, Sigma-Aldrich, NSW, Australia), EC supplement with 4-methylumbelliferyl-β-D-glucuronide (EC-Mug, Sigma-Aldrich, NSW, Australia) and tryptic soy broth supplemented with novobiocin (TSBn, Sigma-Aldrich, NSW, Australia).

    Techniques:

    Sessile cell counts of mono- and mixed five-species mature biofilms. Pathogens were L. monocytogenes (BZ001) or S. aureus (Sa30). The mature biofilms were a 25-h biofilm and a 72-h biofilm for the S. aureus community and the L. monocytogenes community, respectively. Pathogen cell count was assessed on Baird Parker agar for S. aureus and Oxford agar for L. monocytogenes . Total community cell count was assessed on BHI agar. Error bars represent the standard deviations for three biological replicates.

    Journal: Applied and Environmental Microbiology

    Article Title: Behavior of Foodborne Pathogens Listeria monocytogenes and Staphylococcus aureus in Mixed-Species Biofilms Exposed to Biocides

    doi: 10.1128/AEM.02038-18

    Figure Lengend Snippet: Sessile cell counts of mono- and mixed five-species mature biofilms. Pathogens were L. monocytogenes (BZ001) or S. aureus (Sa30). The mature biofilms were a 25-h biofilm and a 72-h biofilm for the S. aureus community and the L. monocytogenes community, respectively. Pathogen cell count was assessed on Baird Parker agar for S. aureus and Oxford agar for L. monocytogenes . Total community cell count was assessed on BHI agar. Error bars represent the standard deviations for three biological replicates.

    Article Snippet: S. aureus cells were enumerated on Baird Parker agar (Oxoid, UK) with egg yolk emulsion (Oxoid, UK), and L. monocytogenes cells were counted on Oxford (Oxoid, UK) with modified Listeria selective supplement (Oxoid, UK).

    Techniques: Cell Counting

    Survival of sessile cells in biofilms formed on SSCs after peracetic acid exposure. For each pathogen, S. aureus (Sa30) (A) or L. monocytogenes (BZ001) (B), mono- and mixed five-species biofilms were developed on SSCs as mature biofilms before being treated with different concentrations of peracetic acid (PA) for 20 min. Sa30, monobiofilm of S. aureus ; Sa30-comSa, mixed five-species biofilms containing Sa30; BZ001, monobiofilm of L. monocytogenes ; BZ001-comLm, mixed five-species biofilms containing BZ001. Results are presented as the means for biological triplicates.

    Journal: Applied and Environmental Microbiology

    Article Title: Behavior of Foodborne Pathogens Listeria monocytogenes and Staphylococcus aureus in Mixed-Species Biofilms Exposed to Biocides

    doi: 10.1128/AEM.02038-18

    Figure Lengend Snippet: Survival of sessile cells in biofilms formed on SSCs after peracetic acid exposure. For each pathogen, S. aureus (Sa30) (A) or L. monocytogenes (BZ001) (B), mono- and mixed five-species biofilms were developed on SSCs as mature biofilms before being treated with different concentrations of peracetic acid (PA) for 20 min. Sa30, monobiofilm of S. aureus ; Sa30-comSa, mixed five-species biofilms containing Sa30; BZ001, monobiofilm of L. monocytogenes ; BZ001-comLm, mixed five-species biofilms containing BZ001. Results are presented as the means for biological triplicates.

    Article Snippet: S. aureus cells were enumerated on Baird Parker agar (Oxoid, UK) with egg yolk emulsion (Oxoid, UK), and L. monocytogenes cells were counted on Oxford (Oxoid, UK) with modified Listeria selective supplement (Oxoid, UK).

    Techniques:

    Survival of sessile cells in biofilms formed on SSCs after treatment with different concentrations of chlorhexidine digluconate for 20 min. For each pathogen, S. aureus (Sa30) (top) or L. monocytogenes (BZ001) (bottom), mono- and mixed five-species biofilms were developed on SSCs as mature biofilms before treatment. Sa30, monobiofilm of S. aureus ; Sa30-comSa, mixed five-species biofilms containing Sa30; BZ001, monobiofilm of L. monocytogenes ; BZ001-comLm, mixed five-species biofilms containing BZ001. Results are presented as the means of biological triplicates for L. monocytogenes (bottom) and only one biological replicate for S. aureus (top).

    Journal: Applied and Environmental Microbiology

    Article Title: Behavior of Foodborne Pathogens Listeria monocytogenes and Staphylococcus aureus in Mixed-Species Biofilms Exposed to Biocides

    doi: 10.1128/AEM.02038-18

    Figure Lengend Snippet: Survival of sessile cells in biofilms formed on SSCs after treatment with different concentrations of chlorhexidine digluconate for 20 min. For each pathogen, S. aureus (Sa30) (top) or L. monocytogenes (BZ001) (bottom), mono- and mixed five-species biofilms were developed on SSCs as mature biofilms before treatment. Sa30, monobiofilm of S. aureus ; Sa30-comSa, mixed five-species biofilms containing Sa30; BZ001, monobiofilm of L. monocytogenes ; BZ001-comLm, mixed five-species biofilms containing BZ001. Results are presented as the means of biological triplicates for L. monocytogenes (bottom) and only one biological replicate for S. aureus (top).

    Article Snippet: S. aureus cells were enumerated on Baird Parker agar (Oxoid, UK) with egg yolk emulsion (Oxoid, UK), and L. monocytogenes cells were counted on Oxford (Oxoid, UK) with modified Listeria selective supplement (Oxoid, UK).

    Techniques:

    Quantification of viable sessile cells in dual-species biofilms. Each pathogen, L. monocytogenes (BZ001) (top) or S. aureus (BZ012) (bottom), was grown as a monofilm or dual biofilm developed on SSCs with each of the associated community members. At least three biological replicates for L. monocytogenes and only duplicates for S. aureus were done.

    Journal: Applied and Environmental Microbiology

    Article Title: Behavior of Foodborne Pathogens Listeria monocytogenes and Staphylococcus aureus in Mixed-Species Biofilms Exposed to Biocides

    doi: 10.1128/AEM.02038-18

    Figure Lengend Snippet: Quantification of viable sessile cells in dual-species biofilms. Each pathogen, L. monocytogenes (BZ001) (top) or S. aureus (BZ012) (bottom), was grown as a monofilm or dual biofilm developed on SSCs with each of the associated community members. At least three biological replicates for L. monocytogenes and only duplicates for S. aureus were done.

    Article Snippet: S. aureus cells were enumerated on Baird Parker agar (Oxoid, UK) with egg yolk emulsion (Oxoid, UK), and L. monocytogenes cells were counted on Oxford (Oxoid, UK) with modified Listeria selective supplement (Oxoid, UK).

    Techniques:

    Monocytes and neutrophils are recruited to sites of L. monocytogenes infection

    Journal: Journal of immunology (Baltimore, Md. : 1950)

    Article Title: Ly6G+ neutrophils are dispensable for defense against systemic Listeria monocytogenes infection

    doi: 10.4049/jimmunol.1101721

    Figure Lengend Snippet: Monocytes and neutrophils are recruited to sites of L. monocytogenes infection

    Article Snippet: Foci of infection were visualized by staining with L. monocytogenes antiserum (BD Biosciences, San Jose, CA), followed by staining with Alexa633-F(ab′)2-anti-rabbit IgG (Invitrogen).

    Techniques: Infection

    Inflammatory monocytes but not neutrophils are essential in control L. monocytogenes infection

    Journal: Journal of immunology (Baltimore, Md. : 1950)

    Article Title: Ly6G+ neutrophils are dispensable for defense against systemic Listeria monocytogenes infection

    doi: 10.4049/jimmunol.1101721

    Figure Lengend Snippet: Inflammatory monocytes but not neutrophils are essential in control L. monocytogenes infection

    Article Snippet: Foci of infection were visualized by staining with L. monocytogenes antiserum (BD Biosciences, San Jose, CA), followed by staining with Alexa633-F(ab′)2-anti-rabbit IgG (Invitrogen).

    Techniques: Infection

    Flavin cofactors and flavin cofactor analogs are present in cell lysates of L. monocytogenes .

    Journal: Journal of Bacteriology

    Article Title: Uptake and Metabolism of Antibiotics Roseoflavin and 8-Demethyl-8-Aminoriboflavin in Riboflavin-Auxotrophic Listeria monocytogenes

    doi: 10.1128/JB.00388-16

    Figure Lengend Snippet: Flavin cofactors and flavin cofactor analogs are present in cell lysates of L. monocytogenes .

    Article Snippet: L. monocytogenes was grown in either brain heart infusion (BHI) broth (Becton Dickinson, Heidelberg, Germany) or in a minimal medium ( ).

    Techniques:

    Lmo1329-catalyzed (A and B) and Lmo0728-catalyzed (C and D) reactions. Different flavin substrates (8-demethyl-8-aminoriboflavin [AF], riboflavin [RF], roseoflavin [RoF]) were used for testing the two different Listeria monocytogenes enzymes, Lmo1329

    Journal: Journal of Bacteriology

    Article Title: Uptake and Metabolism of Antibiotics Roseoflavin and 8-Demethyl-8-Aminoriboflavin in Riboflavin-Auxotrophic Listeria monocytogenes

    doi: 10.1128/JB.00388-16

    Figure Lengend Snippet: Lmo1329-catalyzed (A and B) and Lmo0728-catalyzed (C and D) reactions. Different flavin substrates (8-demethyl-8-aminoriboflavin [AF], riboflavin [RF], roseoflavin [RoF]) were used for testing the two different Listeria monocytogenes enzymes, Lmo1329

    Article Snippet: L. monocytogenes was grown in either brain heart infusion (BHI) broth (Becton Dickinson, Heidelberg, Germany) or in a minimal medium ( ).

    Techniques:

    The FMN riboswitch Rli96 of Listeria monocytogenes in vivo is less responsive to FMN than the corresponding ribDG FMN riboswitch from Bacillus subtilis . Specific β-galactosidase (LacZ) activities in cell extracts of different B. subtilis test

    Journal: Journal of Bacteriology

    Article Title: Uptake and Metabolism of Antibiotics Roseoflavin and 8-Demethyl-8-Aminoriboflavin in Riboflavin-Auxotrophic Listeria monocytogenes

    doi: 10.1128/JB.00388-16

    Figure Lengend Snippet: The FMN riboswitch Rli96 of Listeria monocytogenes in vivo is less responsive to FMN than the corresponding ribDG FMN riboswitch from Bacillus subtilis . Specific β-galactosidase (LacZ) activities in cell extracts of different B. subtilis test

    Article Snippet: L. monocytogenes was grown in either brain heart infusion (BHI) broth (Becton Dickinson, Heidelberg, Germany) or in a minimal medium ( ).

    Techniques: In Vivo

    Flavin cofactors and flavin cofactor analogs are present in cell lysates of L. monocytogenes .

    Journal: Journal of Bacteriology

    Article Title: Uptake and Metabolism of Antibiotics Roseoflavin and 8-Demethyl-8-Aminoriboflavin in Riboflavin-Auxotrophic Listeria monocytogenes

    doi: 10.1128/JB.00388-16

    Figure Lengend Snippet: Flavin cofactors and flavin cofactor analogs are present in cell lysates of L. monocytogenes .

    Article Snippet: L. monocytogenes was grown in either brain heart infusion (BHI) broth (Becton Dickinson, Heidelberg, Germany) or in a minimal medium ( ).

    Techniques:

    The riboflavin (RF) analogs roseoflavin (RoF) and 8-demethyl-8-aminoriboflavin (AF) negatively interfere with growth of different Listeria monocytogenes strains. (A) Growth of L. monocytogenes wild type (wt) (left panel) and L. monocytogenes M1 (M1) (right

    Journal: Journal of Bacteriology

    Article Title: Uptake and Metabolism of Antibiotics Roseoflavin and 8-Demethyl-8-Aminoriboflavin in Riboflavin-Auxotrophic Listeria monocytogenes

    doi: 10.1128/JB.00388-16

    Figure Lengend Snippet: The riboflavin (RF) analogs roseoflavin (RoF) and 8-demethyl-8-aminoriboflavin (AF) negatively interfere with growth of different Listeria monocytogenes strains. (A) Growth of L. monocytogenes wild type (wt) (left panel) and L. monocytogenes M1 (M1) (right

    Article Snippet: L. monocytogenes was grown in either brain heart infusion (BHI) broth (Becton Dickinson, Heidelberg, Germany) or in a minimal medium ( ).

    Techniques:

    The FMN riboswitch Rli96 of Listeria monocytogenes is affected by RoFMN but not by AFMN in vitro . A reporter plasmid (pT7-Rli96-luc) producing a transcriptional fusion of Rli96 and the reporter gene luc (coding for firefly luciferase) were used as a DNA

    Journal: Journal of Bacteriology

    Article Title: Uptake and Metabolism of Antibiotics Roseoflavin and 8-Demethyl-8-Aminoriboflavin in Riboflavin-Auxotrophic Listeria monocytogenes

    doi: 10.1128/JB.00388-16

    Figure Lengend Snippet: The FMN riboswitch Rli96 of Listeria monocytogenes is affected by RoFMN but not by AFMN in vitro . A reporter plasmid (pT7-Rli96-luc) producing a transcriptional fusion of Rli96 and the reporter gene luc (coding for firefly luciferase) were used as a DNA

    Article Snippet: L. monocytogenes was grown in either brain heart infusion (BHI) broth (Becton Dickinson, Heidelberg, Germany) or in a minimal medium ( ).

    Techniques: In Vitro, Plasmid Preparation, Luciferase

    The FMN riboswitch Rli96 of Listeria monocytogenes is affected by riboflavin (RF) and roseoflavin (RoF) but not by 8-demethyl-8-aminoriboflavin (AF) in vivo . The specific β-galactosidase (LacZ) activities in cell extracts of the B. subtilis test

    Journal: Journal of Bacteriology

    Article Title: Uptake and Metabolism of Antibiotics Roseoflavin and 8-Demethyl-8-Aminoriboflavin in Riboflavin-Auxotrophic Listeria monocytogenes

    doi: 10.1128/JB.00388-16

    Figure Lengend Snippet: The FMN riboswitch Rli96 of Listeria monocytogenes is affected by riboflavin (RF) and roseoflavin (RoF) but not by 8-demethyl-8-aminoriboflavin (AF) in vivo . The specific β-galactosidase (LacZ) activities in cell extracts of the B. subtilis test

    Article Snippet: L. monocytogenes was grown in either brain heart infusion (BHI) broth (Becton Dickinson, Heidelberg, Germany) or in a minimal medium ( ).

    Techniques: In Vivo

    Listeria monocytogenes Xen32 strains show attenuated virulence in vivo. BALB/cJ mice (n = 10) were intragastrically inoculated with 1 × 10 10 CFU Lmo-Xen32, Lmo-Xen32-mur, Lmo-EGDe-lux or Lmo-EGDe-mur-lux. (A) Serial BLI of 1 representative mouse from one hour until 7 days post infection as described in Methods (see also Additional file 1 : Figure S1 for reference). Colour bar indicates emitted light with 3 or 4 min integration time in photons/s/cm2/sr. un = uninfected control. (B) Survival rates of BALB/cJ mice (n = 10) after infection with 1 × 10 10 CFU Lmo-Xen32, Lmo-Xen32-mur, Lmo-EGDe-lux or Lmo-EGDe-mur-lux. (C) Body weight loss of BALB/cJ mice infected with the same listerial strains and dosage as shown in (A) and (B) . Graphs demonstrated mean in percent (n = 10) with standard error. Statistical significance between murinised (green stars) and non-murinised (yellow stars) Listeria strains are indicated (*p

    Journal: Gut Pathogens

    Article Title: The bioluminescent Listeria monocytogenes strain Xen32 is defective in flagella expression and highly attenuated in orally infected BALB/cJ mice

    doi: 10.1186/1757-4749-5-19

    Figure Lengend Snippet: Listeria monocytogenes Xen32 strains show attenuated virulence in vivo. BALB/cJ mice (n = 10) were intragastrically inoculated with 1 × 10 10 CFU Lmo-Xen32, Lmo-Xen32-mur, Lmo-EGDe-lux or Lmo-EGDe-mur-lux. (A) Serial BLI of 1 representative mouse from one hour until 7 days post infection as described in Methods (see also Additional file 1 : Figure S1 for reference). Colour bar indicates emitted light with 3 or 4 min integration time in photons/s/cm2/sr. un = uninfected control. (B) Survival rates of BALB/cJ mice (n = 10) after infection with 1 × 10 10 CFU Lmo-Xen32, Lmo-Xen32-mur, Lmo-EGDe-lux or Lmo-EGDe-mur-lux. (C) Body weight loss of BALB/cJ mice infected with the same listerial strains and dosage as shown in (A) and (B) . Graphs demonstrated mean in percent (n = 10) with standard error. Statistical significance between murinised (green stars) and non-murinised (yellow stars) Listeria strains are indicated (*p

    Article Snippet: L. monocytogenes Xen32 was purchased from Perkin Elmer (Rodgau, Germany) and genetically modified for expression of a mouse-adapted InlA as previously described [ ].

    Techniques: In Vivo, Mouse Assay, Infection

    L. monocytogenes Xen32 strains are flagella deficient. Transmission electron microscopy of Lmo-Xen32, Lmo-Xen32-mur, Lmo-10403S, Lmo-EGDe-lux, Lmo-EGDe-mur-lux and Lmo-EGDe. Scale bars indicate 1 μm.

    Journal: Gut Pathogens

    Article Title: The bioluminescent Listeria monocytogenes strain Xen32 is defective in flagella expression and highly attenuated in orally infected BALB/cJ mice

    doi: 10.1186/1757-4749-5-19

    Figure Lengend Snippet: L. monocytogenes Xen32 strains are flagella deficient. Transmission electron microscopy of Lmo-Xen32, Lmo-Xen32-mur, Lmo-10403S, Lmo-EGDe-lux, Lmo-EGDe-mur-lux and Lmo-EGDe. Scale bars indicate 1 μm.

    Article Snippet: L. monocytogenes Xen32 was purchased from Perkin Elmer (Rodgau, Germany) and genetically modified for expression of a mouse-adapted InlA as previously described [ ].

    Techniques: Transmission Assay, Electron Microscopy

    Dynamic of thermal inactivation of L. monocytogenes during traditional cooking process of a naturally contaminated kebab.

    Journal: Italian Journal of Food Safety

    Article Title: Kebab: can the traditional cooking process sanitize a natural contamination by Listeria monocytogenes?

    doi: 10.4081/ijfs.2018.7167

    Figure Lengend Snippet: Dynamic of thermal inactivation of L. monocytogenes during traditional cooking process of a naturally contaminated kebab.

    Article Snippet: For all kebab samples (raw kebab, sliced cooked kebab, kebab that remained in the collecting pan and panini ready to be served with sauces and vegetables) several microbiological parameters were investigated with an accredited laboratory according to ISO 17025:2005: enumeration of MAB (mesophilic aerobic bacteria) (ISO 4833-2:2013/Cor1: ), enumeration of Enterobacteriaceae (ISO 21528-2:2004), enumeration of Beta-glucuronidase positive E.coli (ISO 16649-2:2001), enumeration of LAB (lactic acid bacteria) (MRS agar, 37°C and M17 agar, 37°C±1°C in microaerophilic condition 5% of CO2 ) enumeration of sulfite-reducing bacteria growing under anaerobic conditions (ISO 15213:2003), enumeration of yeasts and molds (OGYEA agar, 21±1°C), and research of Salmonella spp. (AFNOR 2008 + ISO 6579:2002/Cor 1: ), research and enumeration of L. monocytogenes (AFNOR 2009 – 04/05 + ISO 11290-1:1996/Amd 1:2004 and ISO 11290-2:1998/Amd 1:2004), research of shiga toxin-producing Escherichia coli (ISO/TS 13136:2012) and research of Campylobacter spp. (REALTIME - IQ-CHECKTM CAMPYLOBACTER KIT (BIO-RAD) + ISO 10272-1:2006).

    Techniques:

    Locus gtcA . (A) Schematic organization of the gtcA region. (Upper part) rpmE-rho region of B. subtilis . The rpmE is shaded in dark gray; the rho is shaded in light gray. (Middle part) rpmE-gtcA-rho region of L. monocytogenes serotype 4b. (Lower part) rpmE-csbB-gtcA-rho region of L. monocytogenes ). Similar residues are shaded, and identical residues are boxed (thick lines). The numbers refer to amino acid length.

    Journal: Infection and Immunity

    Article Title: Identification of New Genes Involved in the Virulence of Listeria monocytogenes by Signature-Tagged Transposon Mutagenesis

    doi: 10.1128/IAI.69.4.2054-2065.2001

    Figure Lengend Snippet: Locus gtcA . (A) Schematic organization of the gtcA region. (Upper part) rpmE-rho region of B. subtilis . The rpmE is shaded in dark gray; the rho is shaded in light gray. (Middle part) rpmE-gtcA-rho region of L. monocytogenes serotype 4b. (Lower part) rpmE-csbB-gtcA-rho region of L. monocytogenes ). Similar residues are shaded, and identical residues are boxed (thick lines). The numbers refer to amino acid length.

    Article Snippet: The clones from each bank in L. monocytogenes were screened for hemolytic phenotype onto horse blood agar plates (BioMerieux).

    Techniques:

    Locus celR . (Upper part) Schematic organization of the celR locus in B. subtilis . The three domains comprised within the FruA polypeptide are indicated (IIA, IIB, and IIC). (Lower part) Schematic organization of the celR locus in L. monocytogenes . The three genes downstream of celR determine three ORFs that share 34, 44, and 52% identities, respectively, with the corresponding domains of the FruA protein of B. subtilis . The arrows indicate the approximate size and orientation of the different genes.

    Journal: Infection and Immunity

    Article Title: Identification of New Genes Involved in the Virulence of Listeria monocytogenes by Signature-Tagged Transposon Mutagenesis

    doi: 10.1128/IAI.69.4.2054-2065.2001

    Figure Lengend Snippet: Locus celR . (Upper part) Schematic organization of the celR locus in B. subtilis . The three domains comprised within the FruA polypeptide are indicated (IIA, IIB, and IIC). (Lower part) Schematic organization of the celR locus in L. monocytogenes . The three genes downstream of celR determine three ORFs that share 34, 44, and 52% identities, respectively, with the corresponding domains of the FruA protein of B. subtilis . The arrows indicate the approximate size and orientation of the different genes.

    Article Snippet: The clones from each bank in L. monocytogenes were screened for hemolytic phenotype onto horse blood agar plates (BioMerieux).

    Techniques:

    Relative transcript expression profiles of select genes from weakly-adherent (CW35) and strongly-adherent (99-38) strains of L. monocytogenes . Panel A, from cells recovered from planktonic growth at 30 °C. Panel B, from cells attached to glass beads during growth at 30 °C. Panel C, from planktonic cells grown at 42 °C. Expression is relative to that of the reference gene, 16S rRNA. All data bars represent the means of triplicate replications for gene expression RT-qPCR assays. Error bars indicate the standard deviation from the mean. Expression was normalized (×10 7 factor) to eliminate negative expression levels.

    Journal: Pathogens

    Article Title: RT-qPCR Analysis of 15 Genes Encoding Putative Surface Proteins Involved in Adherence of Listeria monocytogenes

    doi: 10.3390/pathogens5040060

    Figure Lengend Snippet: Relative transcript expression profiles of select genes from weakly-adherent (CW35) and strongly-adherent (99-38) strains of L. monocytogenes . Panel A, from cells recovered from planktonic growth at 30 °C. Panel B, from cells attached to glass beads during growth at 30 °C. Panel C, from planktonic cells grown at 42 °C. Expression is relative to that of the reference gene, 16S rRNA. All data bars represent the means of triplicate replications for gene expression RT-qPCR assays. Error bars indicate the standard deviation from the mean. Expression was normalized (×10 7 factor) to eliminate negative expression levels.

    Article Snippet: Planktonic Cells Pelleted cells of various strains of L. monocytogenes in sterile Eppendorf tubes were prepared from 1 mL of overnight cultures in BHI broth at 30 °C or 42 °C, and washed 3 times by suspension with 1× PBS prior to total RNA extraction.

    Techniques: Expressing, Quantitative RT-PCR, Standard Deviation

    Comparison of attachment characteristics of L. monocytogenes CW35 (weakly-adherent) and 99-38 (strongly-adherent) in microplate wells. Enumeration of well cell cultures (left) and attached cells (right) after release by treatment with protease. All data represent the means of triplicate replications. Means with the same lowercase letters are not significantly different; means with different letters are significantly different ( P

    Journal: Pathogens

    Article Title: RT-qPCR Analysis of 15 Genes Encoding Putative Surface Proteins Involved in Adherence of Listeria monocytogenes

    doi: 10.3390/pathogens5040060

    Figure Lengend Snippet: Comparison of attachment characteristics of L. monocytogenes CW35 (weakly-adherent) and 99-38 (strongly-adherent) in microplate wells. Enumeration of well cell cultures (left) and attached cells (right) after release by treatment with protease. All data represent the means of triplicate replications. Means with the same lowercase letters are not significantly different; means with different letters are significantly different ( P

    Article Snippet: Planktonic Cells Pelleted cells of various strains of L. monocytogenes in sterile Eppendorf tubes were prepared from 1 mL of overnight cultures in BHI broth at 30 °C or 42 °C, and washed 3 times by suspension with 1× PBS prior to total RNA extraction.

    Techniques:

    PCR products from genomic DNA of L. monocytogenes EDGe (Panel A), 99-38 (Panel B), and CW35 (Panel C) for PCR nucleotide evaluation of lmo0723, lmo1068, lmo1076, and lmo2558. Different gene-specific primer pairs were used for PCR amplification and subsequent agarose gel analysis of products. PCR primer combinations were based on L. monocytogenes type strain EGDe (Panel A) and tested on 99-38 (Panel B) and CW35 (Panel C). Gene lmo0723 : Lane 1, 0723A (148bp); 2, 0723B (416bp); 3, 0723C (150bp); 4, 0723D (505bp); lmo1068 : 5, 1068A (149bp); 6, 1068B (438bp); 7, 1068C (149bp); 8, 1068D (440bp); 9, 1068E (147bp); lmo1076 : 10, 1076A (470bp); 11, 1076B (150bp); 12, 1076C (146bp); 13, 1076D (991bp); lmo2558 : 14, 2558A (458bp); 15, 2558B (148bp); 16, 2558C (149bp); 17, 2558D (1129bp); 18, 100bp DNA ladder; 19 and 20, positive controls.

    Journal: Pathogens

    Article Title: RT-qPCR Analysis of 15 Genes Encoding Putative Surface Proteins Involved in Adherence of Listeria monocytogenes

    doi: 10.3390/pathogens5040060

    Figure Lengend Snippet: PCR products from genomic DNA of L. monocytogenes EDGe (Panel A), 99-38 (Panel B), and CW35 (Panel C) for PCR nucleotide evaluation of lmo0723, lmo1068, lmo1076, and lmo2558. Different gene-specific primer pairs were used for PCR amplification and subsequent agarose gel analysis of products. PCR primer combinations were based on L. monocytogenes type strain EGDe (Panel A) and tested on 99-38 (Panel B) and CW35 (Panel C). Gene lmo0723 : Lane 1, 0723A (148bp); 2, 0723B (416bp); 3, 0723C (150bp); 4, 0723D (505bp); lmo1068 : 5, 1068A (149bp); 6, 1068B (438bp); 7, 1068C (149bp); 8, 1068D (440bp); 9, 1068E (147bp); lmo1076 : 10, 1076A (470bp); 11, 1076B (150bp); 12, 1076C (146bp); 13, 1076D (991bp); lmo2558 : 14, 2558A (458bp); 15, 2558B (148bp); 16, 2558C (149bp); 17, 2558D (1129bp); 18, 100bp DNA ladder; 19 and 20, positive controls.

    Article Snippet: Planktonic Cells Pelleted cells of various strains of L. monocytogenes in sterile Eppendorf tubes were prepared from 1 mL of overnight cultures in BHI broth at 30 °C or 42 °C, and washed 3 times by suspension with 1× PBS prior to total RNA extraction.

    Techniques: Polymerase Chain Reaction, Amplification, Agarose Gel Electrophoresis

    Adherence of various strains of L. monocytogenes using the microplate fluorescence (5,6-CFDA) adherence assay. Weakly- and strongly-adherent strains are represented by black and red bars, respectively. Data bars represent the mean of triplicate replications. Means that share the same lowercase letters are not significantly different; means with different letters are significantly different ( P

    Journal: Pathogens

    Article Title: RT-qPCR Analysis of 15 Genes Encoding Putative Surface Proteins Involved in Adherence of Listeria monocytogenes

    doi: 10.3390/pathogens5040060

    Figure Lengend Snippet: Adherence of various strains of L. monocytogenes using the microplate fluorescence (5,6-CFDA) adherence assay. Weakly- and strongly-adherent strains are represented by black and red bars, respectively. Data bars represent the mean of triplicate replications. Means that share the same lowercase letters are not significantly different; means with different letters are significantly different ( P

    Article Snippet: Planktonic Cells Pelleted cells of various strains of L. monocytogenes in sterile Eppendorf tubes were prepared from 1 mL of overnight cultures in BHI broth at 30 °C or 42 °C, and washed 3 times by suspension with 1× PBS prior to total RNA extraction.

    Techniques: Fluorescence

    Effect of temperature (30 °C vs. 42 °C) on attachment of different adherence-variant strains of L. monocytogenes (strongly adherent: Jag167, 99-38, EGDe; weakly adherent: CW35, CW52) as determined by the microplate adherence assay. Uninoculated brain heart infusion (BHI) nutrient broth was tested as a control. All data represent the means of triplicate replications. Means with the same upper/lowercase letters are not significantly different; means with different upper/lower case letters are significantly different ( P

    Journal: Pathogens

    Article Title: RT-qPCR Analysis of 15 Genes Encoding Putative Surface Proteins Involved in Adherence of Listeria monocytogenes

    doi: 10.3390/pathogens5040060

    Figure Lengend Snippet: Effect of temperature (30 °C vs. 42 °C) on attachment of different adherence-variant strains of L. monocytogenes (strongly adherent: Jag167, 99-38, EGDe; weakly adherent: CW35, CW52) as determined by the microplate adherence assay. Uninoculated brain heart infusion (BHI) nutrient broth was tested as a control. All data represent the means of triplicate replications. Means with the same upper/lowercase letters are not significantly different; means with different upper/lower case letters are significantly different ( P

    Article Snippet: Planktonic Cells Pelleted cells of various strains of L. monocytogenes in sterile Eppendorf tubes were prepared from 1 mL of overnight cultures in BHI broth at 30 °C or 42 °C, and washed 3 times by suspension with 1× PBS prior to total RNA extraction.

    Techniques: Variant Assay

    ELISA O-antigen and H-antigen reactions of serogroup 1/2 and 3 L. monocytogenes strains. (A) Strain J0098, serotype 1/2a; (B) strain G848, serotype 1/2b; (C) strain G-3321, serotype 1/2c; (D) strain J0095, serotype 3a; (E) strain cpp81, serotype 3b; (F) strain J0096, serotype 3c.

    Journal: Journal of Clinical Microbiology

    Article Title: Serotyping of Listeria monocytogenes by Enzyme-Linked Immunosorbent Assay and Identification of Mixed-Serotype Cultures by Colony Immunoblotting

    doi: 10.1128/JCM.41.2.564-571.2003

    Figure Lengend Snippet: ELISA O-antigen and H-antigen reactions of serogroup 1/2 and 3 L. monocytogenes strains. (A) Strain J0098, serotype 1/2a; (B) strain G848, serotype 1/2b; (C) strain G-3321, serotype 1/2c; (D) strain J0095, serotype 3a; (E) strain cpp81, serotype 3b; (F) strain J0096, serotype 3c.

    Article Snippet: Isolates were confirmed as L. monocytogenes by production of turquoise colonies on BCM L. monocytogenes plating medium (Biosynth International, Naperville, Ill.) ( ) and by PCR amplification of an iap gene fragment using L. monocytogenes -specific primers ( ).

    Techniques: Enzyme-linked Immunosorbent Assay

    Colony immunoblotting of single- and mixed-serotype cultures of L. monocytogenes . Blots of serotype 1/2a strain J0098 (A), serotype 4b strain J0097 (B), or a mixture of strains J0098 and J0097 (C) were probed sequentially with O-factor antiserum I/II (stained red) and O-factor antiserum V/VI (stained green). (D) A streak plate of strain 19118 was blotted and probed sequentially with O-factor antiserum V/VI (stained red) and O-factor antiserum I/II (stained green). Insets are magnifications (×5) of the indicated region of each filter to show detail.

    Journal: Journal of Clinical Microbiology

    Article Title: Serotyping of Listeria monocytogenes by Enzyme-Linked Immunosorbent Assay and Identification of Mixed-Serotype Cultures by Colony Immunoblotting

    doi: 10.1128/JCM.41.2.564-571.2003

    Figure Lengend Snippet: Colony immunoblotting of single- and mixed-serotype cultures of L. monocytogenes . Blots of serotype 1/2a strain J0098 (A), serotype 4b strain J0097 (B), or a mixture of strains J0098 and J0097 (C) were probed sequentially with O-factor antiserum I/II (stained red) and O-factor antiserum V/VI (stained green). (D) A streak plate of strain 19118 was blotted and probed sequentially with O-factor antiserum V/VI (stained red) and O-factor antiserum I/II (stained green). Insets are magnifications (×5) of the indicated region of each filter to show detail.

    Article Snippet: Isolates were confirmed as L. monocytogenes by production of turquoise colonies on BCM L. monocytogenes plating medium (Biosynth International, Naperville, Ill.) ( ) and by PCR amplification of an iap gene fragment using L. monocytogenes -specific primers ( ).

    Techniques: Staining

    Recretion of selected InlA mutations in EGD-e . A . Comparison of the invasion attributes of EGD-e and EGD-e InlA m * (Ser192Asn/Trp369Ser). Exponential phase L. monocytogenes cells (OD = 0.8) were invaded (MOI of 25:1) in triplicate for 1 h before overlaying with gentamicin. Invasion was expressed as the average cfu count per well (with standard deviation) or invasion relative to EGD-e (below graph). The graph is representative of the data from three independent experiments. B . The relative virulence of EGD-e compared against EGD-e InlA m * (tagged with pIMC3 kan and pIMC3 ery respectively) was accessed by competitive index after i.v. infection (1 × 10 4 cfu of each strain) of 15 Balb/c mice. On each subsequent day 5 mice were euthanized and spleens and livers aseptically removed and enumerated. Data are presented as the mean and standard deviation of 5 mice, competitive indices and statistical analysis was conducted using the one sample t test as described previously [ 18 ]. NS = Not significant. C . Oral inoculations of Balb/c mice with EGD-e::pIMC3 kan and EGD-e InlA m * ::pIMC ery mixed at a 1:1 ratio in a total inoculum of 1 × 10 10 cfu/200 μl containing 100 mg of CaCO 3 . *** = p

    Journal: BMC Microbiology

    Article Title: Directed evolution and targeted mutagenesis to murinize listeria monocytogenes internalin A for enhanced infectivity in the murine oral infection model

    doi: 10.1186/1471-2180-10-318

    Figure Lengend Snippet: Recretion of selected InlA mutations in EGD-e . A . Comparison of the invasion attributes of EGD-e and EGD-e InlA m * (Ser192Asn/Trp369Ser). Exponential phase L. monocytogenes cells (OD = 0.8) were invaded (MOI of 25:1) in triplicate for 1 h before overlaying with gentamicin. Invasion was expressed as the average cfu count per well (with standard deviation) or invasion relative to EGD-e (below graph). The graph is representative of the data from three independent experiments. B . The relative virulence of EGD-e compared against EGD-e InlA m * (tagged with pIMC3 kan and pIMC3 ery respectively) was accessed by competitive index after i.v. infection (1 × 10 4 cfu of each strain) of 15 Balb/c mice. On each subsequent day 5 mice were euthanized and spleens and livers aseptically removed and enumerated. Data are presented as the mean and standard deviation of 5 mice, competitive indices and statistical analysis was conducted using the one sample t test as described previously [ 18 ]. NS = Not significant. C . Oral inoculations of Balb/c mice with EGD-e::pIMC3 kan and EGD-e InlA m * ::pIMC ery mixed at a 1:1 ratio in a total inoculum of 1 × 10 10 cfu/200 μl containing 100 mg of CaCO 3 . *** = p

    Article Snippet: The discrepancy is due to a glutamate at residue 16 in mouse (and rat) E-cadherin rendering these host species relatively resistant to infection by the oral route and limiting their use as laboratory models for certain L. monocytogenes- mediated disease processes [ ].

    Techniques: Standard Deviation, Infection, Mouse Assay

    InlA dependent invasion of EGD-e derrived strains into human (Caco-2: grey bars) or murine (CT-26: white bars) monolayers . Exponential phase L. monocytogenes cells (OD = 0.8) were invaded (MOI of 25:1) in triplicate for 1 h before overlaying with gentamicin. Invasion was expressed as the average cfu count per well (with standard deviation) or invasion relative to EGD-e (below graph) (n = 3). The graph is representative of the data from three independent experiments.

    Journal: BMC Microbiology

    Article Title: Directed evolution and targeted mutagenesis to murinize listeria monocytogenes internalin A for enhanced infectivity in the murine oral infection model

    doi: 10.1186/1471-2180-10-318

    Figure Lengend Snippet: InlA dependent invasion of EGD-e derrived strains into human (Caco-2: grey bars) or murine (CT-26: white bars) monolayers . Exponential phase L. monocytogenes cells (OD = 0.8) were invaded (MOI of 25:1) in triplicate for 1 h before overlaying with gentamicin. Invasion was expressed as the average cfu count per well (with standard deviation) or invasion relative to EGD-e (below graph) (n = 3). The graph is representative of the data from three independent experiments.

    Article Snippet: The discrepancy is due to a glutamate at residue 16 in mouse (and rat) E-cadherin rendering these host species relatively resistant to infection by the oral route and limiting their use as laboratory models for certain L. monocytogenes- mediated disease processes [ ].

    Techniques: Standard Deviation

    Invasion attributes of individual L. lactis clones post CT-26 enrichment (passage 6) into Caco-2 (grey bars) or CT-26 (white bars) cells . From each of the four banks, eight clones were picked and invaded with invasion expressed as the average (with standard deviation) from triplicate wells. Sequnce data of the clones is presented in Table 2. Letters above bars indicate sequences that were subsequently used to recreate into the L. monocytogenes chromosome. The controls InlA WT (WT) and InlA m * ( mur ) expressing L. lactis were included for comparison. The graph is of the data from one experiment.

    Journal: BMC Microbiology

    Article Title: Directed evolution and targeted mutagenesis to murinize listeria monocytogenes internalin A for enhanced infectivity in the murine oral infection model

    doi: 10.1186/1471-2180-10-318

    Figure Lengend Snippet: Invasion attributes of individual L. lactis clones post CT-26 enrichment (passage 6) into Caco-2 (grey bars) or CT-26 (white bars) cells . From each of the four banks, eight clones were picked and invaded with invasion expressed as the average (with standard deviation) from triplicate wells. Sequnce data of the clones is presented in Table 2. Letters above bars indicate sequences that were subsequently used to recreate into the L. monocytogenes chromosome. The controls InlA WT (WT) and InlA m * ( mur ) expressing L. lactis were included for comparison. The graph is of the data from one experiment.

    Article Snippet: The discrepancy is due to a glutamate at residue 16 in mouse (and rat) E-cadherin rendering these host species relatively resistant to infection by the oral route and limiting their use as laboratory models for certain L. monocytogenes- mediated disease processes [ ].

    Techniques: Clone Assay, Standard Deviation, Expressing

    Influence of pG, GO, and rGO on the growth of Listeria monocytogenes and Salmonella enterica at 25 and 250 μg/mL. Data presented are the average of triplicate determinations, with error bars representing mean standard error.

    Journal: Nanoscale Research Letters

    Article Title: Interaction of graphene family materials with Listeria monocytogenes and Salmonella enterica

    doi: 10.1186/s11671-015-0749-y

    Figure Lengend Snippet: Influence of pG, GO, and rGO on the growth of Listeria monocytogenes and Salmonella enterica at 25 and 250 μg/mL. Data presented are the average of triplicate determinations, with error bars representing mean standard error.

    Article Snippet: L. monocytogenes increased the zeta potential of pG and rGO, whereas S. enterica decreased the zeta potential of GO and rGO.

    Techniques:

    Effect of 25 μg/mL concentration of pG, GO, and rGO on the zeta potential of L . monocytogenes (L) and S . enterica (S). Data presented are the average of triplicate determinations, with error bars representing mean standard error.

    Journal: Nanoscale Research Letters

    Article Title: Interaction of graphene family materials with Listeria monocytogenes and Salmonella enterica

    doi: 10.1186/s11671-015-0749-y

    Figure Lengend Snippet: Effect of 25 μg/mL concentration of pG, GO, and rGO on the zeta potential of L . monocytogenes (L) and S . enterica (S). Data presented are the average of triplicate determinations, with error bars representing mean standard error.

    Article Snippet: L. monocytogenes increased the zeta potential of pG and rGO, whereas S. enterica decreased the zeta potential of GO and rGO.

    Techniques: Concentration Assay

    Visualization of the interaction of graphene family materials with Listeria monocytogenes using transmission electron microscopy. Pristine graphene (A, D, G) , graphene oxide ( B, E, H) , and reduced graphene oxide (C, F, I) . Black arrows indicate the graphene material and white arrows the bacterial cells.

    Journal: Nanoscale Research Letters

    Article Title: Interaction of graphene family materials with Listeria monocytogenes and Salmonella enterica

    doi: 10.1186/s11671-015-0749-y

    Figure Lengend Snippet: Visualization of the interaction of graphene family materials with Listeria monocytogenes using transmission electron microscopy. Pristine graphene (A, D, G) , graphene oxide ( B, E, H) , and reduced graphene oxide (C, F, I) . Black arrows indicate the graphene material and white arrows the bacterial cells.

    Article Snippet: L. monocytogenes increased the zeta potential of pG and rGO, whereas S. enterica decreased the zeta potential of GO and rGO.

    Techniques: Transmission Assay, Electron Microscopy

    In IL-17 Receptor Deficient Mice, M. Pulmonis Infection Does Not Facilitate Clearance of L. Monocytogenes or Increase Gr-1+CD11b+ Cells. At day 17 p.i. with M. pulmonis or broth, C57BL/6 and IL-17 receptor deficient mice were infected with L. monocytogenes

    Journal: European journal of immunology

    Article Title: A Novel IL-17 Dependent Mechanism of Cross Protection: Respiratory Infection with Mycoplasma Protects Against a Secondary Listeria Infection

    doi: 10.1002/eji.200838726

    Figure Lengend Snippet: In IL-17 Receptor Deficient Mice, M. Pulmonis Infection Does Not Facilitate Clearance of L. Monocytogenes or Increase Gr-1+CD11b+ Cells. At day 17 p.i. with M. pulmonis or broth, C57BL/6 and IL-17 receptor deficient mice were infected with L. monocytogenes

    Article Snippet: At day 3 p.i. with L. monocytogenes , whole lung homogenates and sera were collected to analyze IL-17 with the Luminex assay.

    Techniques: Mouse Assay, Infection

    Gr-1+CD11b+ Cells Increase During Co-Infection with L. Monocytogenes and M. Pulmonis . At day 17 p.i. with M. pulmonis , C57BL/6 mice were inoculated with 5×10 4 CFUs of L. monocytogenes or PBS. At day 3 p.i. with L. monocytogenes , splenocytes were

    Journal: European journal of immunology

    Article Title: A Novel IL-17 Dependent Mechanism of Cross Protection: Respiratory Infection with Mycoplasma Protects Against a Secondary Listeria Infection

    doi: 10.1002/eji.200838726

    Figure Lengend Snippet: Gr-1+CD11b+ Cells Increase During Co-Infection with L. Monocytogenes and M. Pulmonis . At day 17 p.i. with M. pulmonis , C57BL/6 mice were inoculated with 5×10 4 CFUs of L. monocytogenes or PBS. At day 3 p.i. with L. monocytogenes , splenocytes were

    Article Snippet: At day 3 p.i. with L. monocytogenes , whole lung homogenates and sera were collected to analyze IL-17 with the Luminex assay.

    Techniques: Infection, Mouse Assay

    Infection with M. Pulmonis Increases IL-17, but not IFN-γ, Levels in L. Monocytogenes Infected Mice. At day 17 p.i. with M. pulmonis or broth, C57BL/6 mice were infected with L. monocytogenes or PBS. The concentration of IL-17 in lung homogenates

    Journal: European journal of immunology

    Article Title: A Novel IL-17 Dependent Mechanism of Cross Protection: Respiratory Infection with Mycoplasma Protects Against a Secondary Listeria Infection

    doi: 10.1002/eji.200838726

    Figure Lengend Snippet: Infection with M. Pulmonis Increases IL-17, but not IFN-γ, Levels in L. Monocytogenes Infected Mice. At day 17 p.i. with M. pulmonis or broth, C57BL/6 mice were infected with L. monocytogenes or PBS. The concentration of IL-17 in lung homogenates

    Article Snippet: At day 3 p.i. with L. monocytogenes , whole lung homogenates and sera were collected to analyze IL-17 with the Luminex assay.

    Techniques: Infection, Mouse Assay, Concentration Assay

    Granulocyte Depletion Diminishes Resistance to L. Monocytogenes Conferred by M. Pulmonis . At day 16 p.i. with M. pulmonis or broth, mice were injected with the control antibody (A) or Gr-1 depleting antibody (B). Twenty-four hrs following depletion, mice

    Journal: European journal of immunology

    Article Title: A Novel IL-17 Dependent Mechanism of Cross Protection: Respiratory Infection with Mycoplasma Protects Against a Secondary Listeria Infection

    doi: 10.1002/eji.200838726

    Figure Lengend Snippet: Granulocyte Depletion Diminishes Resistance to L. Monocytogenes Conferred by M. Pulmonis . At day 16 p.i. with M. pulmonis or broth, mice were injected with the control antibody (A) or Gr-1 depleting antibody (B). Twenty-four hrs following depletion, mice

    Article Snippet: At day 3 p.i. with L. monocytogenes , whole lung homogenates and sera were collected to analyze IL-17 with the Luminex assay.

    Techniques: Mouse Assay, Injection

    Prior Infection with M. Pulmonis Confers Resistance Against L. Monocytogenes Infection. C57BL/6 mice were i.v. infected with 1×10 8 (A), 1×10 6 (B), 1×10 4 (C), or 5×10 4 (D, E, F) CFUs of L. monocytogenes (LM) 17 days after

    Journal: European journal of immunology

    Article Title: A Novel IL-17 Dependent Mechanism of Cross Protection: Respiratory Infection with Mycoplasma Protects Against a Secondary Listeria Infection

    doi: 10.1002/eji.200838726

    Figure Lengend Snippet: Prior Infection with M. Pulmonis Confers Resistance Against L. Monocytogenes Infection. C57BL/6 mice were i.v. infected with 1×10 8 (A), 1×10 6 (B), 1×10 4 (C), or 5×10 4 (D, E, F) CFUs of L. monocytogenes (LM) 17 days after

    Article Snippet: At day 3 p.i. with L. monocytogenes , whole lung homogenates and sera were collected to analyze IL-17 with the Luminex assay.

    Techniques: Infection, Mouse Assay

    Schematic outlining the interaction of L. monocytogenes with bile during infection. (A) L. monocytogenes ). (B) Infection of the gallbladder is mediated by an as-yet-undetermined mechanism. Growth in GB bile within the lumen requires specific metabolic processes but is independent of specific resistance mechanisms (the present study).

    Journal: Infection and Immunity

    Article Title: Investigation of the Mechanisms by Which Listeria monocytogenes Grows in Porcine Gallbladder Bile ▿ Grows in Porcine Gallbladder Bile ▿ †

    doi: 10.1128/IAI.00330-10

    Figure Lengend Snippet: Schematic outlining the interaction of L. monocytogenes with bile during infection. (A) L. monocytogenes ). (B) Infection of the gallbladder is mediated by an as-yet-undetermined mechanism. Growth in GB bile within the lumen requires specific metabolic processes but is independent of specific resistance mechanisms (the present study).

    Article Snippet: Molecular systems previously determined to play a role in bile tolerance in L. monocytogenes (BSH, BilE, and Sigma B) were not required for growth in ex vivo GB bile as a growth substrate.

    Techniques: Infection

    Growth of Listeria species in GB bile. (A) Growth of Listeria monocytogenes EGDe (•) and Listeria innocua FH2033 (▵) in ex vivo porcine GB bile. Viable cell counts were carried out at intervals after dilution in one-quarter-strength Ringer's solution and enumeration on BHI. Each time point represents the mean value of at least three independent experiments. (B) Lux-tagged L. monocytogenes strain inoculated into porcine gallbladders imaged under the IVIS system. (C) Scanning electron micrographs of epithelial cells lining the lumen of porcine gallbladder. False colored infected gallbladder shows listerial cells attached to the wall of the lumen. (D) Uninfected gallbladder shows no bacterial cells.

    Journal: Infection and Immunity

    Article Title: Investigation of the Mechanisms by Which Listeria monocytogenes Grows in Porcine Gallbladder Bile ▿ Grows in Porcine Gallbladder Bile ▿ †

    doi: 10.1128/IAI.00330-10

    Figure Lengend Snippet: Growth of Listeria species in GB bile. (A) Growth of Listeria monocytogenes EGDe (•) and Listeria innocua FH2033 (▵) in ex vivo porcine GB bile. Viable cell counts were carried out at intervals after dilution in one-quarter-strength Ringer's solution and enumeration on BHI. Each time point represents the mean value of at least three independent experiments. (B) Lux-tagged L. monocytogenes strain inoculated into porcine gallbladders imaged under the IVIS system. (C) Scanning electron micrographs of epithelial cells lining the lumen of porcine gallbladder. False colored infected gallbladder shows listerial cells attached to the wall of the lumen. (D) Uninfected gallbladder shows no bacterial cells.

    Article Snippet: Molecular systems previously determined to play a role in bile tolerance in L. monocytogenes (BSH, BilE, and Sigma B) were not required for growth in ex vivo GB bile as a growth substrate.

    Techniques: Ex Vivo, Infection

    Growth of specific L. monocytogenes mutants in ex vivo GB bile. L. monocytogenes EGDe and isogenic bilE , bsh , sigB , and prfA deletion mutants show comparable growth in GB bile at pH 7 after 6 h. At the reduced pH of 5.5 the Δ bilE , Δ bsh and Δ sigB mutants are undetectable after a 6-h period, whereas the Δ prfA mutant is comparable to wild-type levels. The broken line is indicative of the initial inoculums. Error bars represent the standard deviations of triplicate experiments. ND, not detected.

    Journal: Infection and Immunity

    Article Title: Investigation of the Mechanisms by Which Listeria monocytogenes Grows in Porcine Gallbladder Bile ▿ Grows in Porcine Gallbladder Bile ▿ †

    doi: 10.1128/IAI.00330-10

    Figure Lengend Snippet: Growth of specific L. monocytogenes mutants in ex vivo GB bile. L. monocytogenes EGDe and isogenic bilE , bsh , sigB , and prfA deletion mutants show comparable growth in GB bile at pH 7 after 6 h. At the reduced pH of 5.5 the Δ bilE , Δ bsh and Δ sigB mutants are undetectable after a 6-h period, whereas the Δ prfA mutant is comparable to wild-type levels. The broken line is indicative of the initial inoculums. Error bars represent the standard deviations of triplicate experiments. ND, not detected.

    Article Snippet: Molecular systems previously determined to play a role in bile tolerance in L. monocytogenes (BSH, BilE, and Sigma B) were not required for growth in ex vivo GB bile as a growth substrate.

    Techniques: Ex Vivo, Mutagenesis

    Fluorescence properties of permeabilized L. monocytogenes EGD-e/pNZ-P help -pHluorin bacteria. (A) In order to determine the appropriate concentration for permeabilization, L. monocytogenes EGD-e was incubated with different concentrations of CTAB (0 - 0.01 %) and stained with Syto ® 9 and propidium iodide (PI) to discriminate intact, live from permeabilized, dead bacteria. Images in the Syto ® 9 and PI channels were aquired with a 100x objective and appropriate filter sets for excitation and emission of the two dyes (Scale bars: 2 μm). (B) L. monocytogenes EGD-e/pNZ-P help -pHluorin was permeabilized in LMB at different pH containing 0.005% CTAB and, at the indicated pH, excitation spectra (350–490 nm) was aquired measuring relative fluorescence intensity (RFI) at 510 nm emission.

    Journal: Frontiers in Microbiology

    Article Title: Intracellular pHluorin as Sensor for Easy Assessment of Bacteriocin-Induced Membrane-Damage in Listeria monocytogenes

    doi: 10.3389/fmicb.2018.03038

    Figure Lengend Snippet: Fluorescence properties of permeabilized L. monocytogenes EGD-e/pNZ-P help -pHluorin bacteria. (A) In order to determine the appropriate concentration for permeabilization, L. monocytogenes EGD-e was incubated with different concentrations of CTAB (0 - 0.01 %) and stained with Syto ® 9 and propidium iodide (PI) to discriminate intact, live from permeabilized, dead bacteria. Images in the Syto ® 9 and PI channels were aquired with a 100x objective and appropriate filter sets for excitation and emission of the two dyes (Scale bars: 2 μm). (B) L. monocytogenes EGD-e/pNZ-P help -pHluorin was permeabilized in LMB at different pH containing 0.005% CTAB and, at the indicated pH, excitation spectra (350–490 nm) was aquired measuring relative fluorescence intensity (RFI) at 510 nm emission.

    Article Snippet: A pHluorin coding sequence, which was codon-optimized for L. monocytogenes ( pHluorinLmo ; Table ) was synthesized and purchased as 752 bp DNA fragment from a commercial service provider (Eurofins Genomics, Ebersberg, Germany) and cloned into pJET1.2 according to manufacturer’s instructions (Thermo Scientific, Waltham, MA, United States).

    Techniques: Fluorescence, Concentration Assay, Incubation, Staining

    Fluorescence properties of pHluorin in crude extracts of L. monocytogenes EGD-e/pNZ-P help -pHluorin. (A) Native (n) or heat-inactivated (hi) crude extracts of L. monocytogenes EGD-e/pNZ-P help -pHluorin and L. monocytogenes EGD-e/pNZ44 were resolved by SDS-PAGE. The image of the gel is an overlay of the channels for pHluorin (excitation at 455–485 nm; emission at 510–555 nm) and protein marker (excitation at 610–635; emission at 675–720 nm). (B) Crude extracts were diluted in LMB and pH was adjusted with 1M HCl. At the indicated pH values, excitation spectra (350–490 nm) were aquired measuring relative fluorescence intensity (RFI) at 510 nm emission.

    Journal: Frontiers in Microbiology

    Article Title: Intracellular pHluorin as Sensor for Easy Assessment of Bacteriocin-Induced Membrane-Damage in Listeria monocytogenes

    doi: 10.3389/fmicb.2018.03038

    Figure Lengend Snippet: Fluorescence properties of pHluorin in crude extracts of L. monocytogenes EGD-e/pNZ-P help -pHluorin. (A) Native (n) or heat-inactivated (hi) crude extracts of L. monocytogenes EGD-e/pNZ-P help -pHluorin and L. monocytogenes EGD-e/pNZ44 were resolved by SDS-PAGE. The image of the gel is an overlay of the channels for pHluorin (excitation at 455–485 nm; emission at 510–555 nm) and protein marker (excitation at 610–635; emission at 675–720 nm). (B) Crude extracts were diluted in LMB and pH was adjusted with 1M HCl. At the indicated pH values, excitation spectra (350–490 nm) were aquired measuring relative fluorescence intensity (RFI) at 510 nm emission.

    Article Snippet: A pHluorin coding sequence, which was codon-optimized for L. monocytogenes ( pHluorinLmo ; Table ) was synthesized and purchased as 752 bp DNA fragment from a commercial service provider (Eurofins Genomics, Ebersberg, Germany) and cloned into pJET1.2 according to manufacturer’s instructions (Thermo Scientific, Waltham, MA, United States).

    Techniques: Fluorescence, SDS Page, Marker

    Assay to determine membrane-damaging activity in supernatants of bacteriocin producing bacteria using L. monocytogenes EGD-e/pNZ-P help -pHluorin. (A) L. monocytogenes EGD-e/pNZ-P help -pHluorin was incubated at pH 6.5 in either LMB (solid bars) or a 1:1 mix of LMB with supernatant of L. lactis MG1363 (LMB:Sup; hatched bars) containing 5 μg/ml nisin. CTAB (0.005%) was used as a positive control (+) and untreated bacteria in either LMB or LMB:Sup without bacteriocin served as negative control (-). (B) OD 600 of L. lactis MG1363 (black) or B1627 (blue) grown in GM17 medium. (C) Supernatants were collected for the cultures shown in (B) at the indicated time points during growth and incubated with L. monocytogenes EGD-e/pNZ-P help -pHluorin (1:1 mix of supernatant with sensor strain in LMB). Values in (A) and (C) are ratios of fluorescence intensities (emission at 510 nm) after excitation at 400 and 470 nm and are mean ± standard deviation of two (A) or four (C) independent experiments using different supernatants and different cultures of the sensor strain.

    Journal: Frontiers in Microbiology

    Article Title: Intracellular pHluorin as Sensor for Easy Assessment of Bacteriocin-Induced Membrane-Damage in Listeria monocytogenes

    doi: 10.3389/fmicb.2018.03038

    Figure Lengend Snippet: Assay to determine membrane-damaging activity in supernatants of bacteriocin producing bacteria using L. monocytogenes EGD-e/pNZ-P help -pHluorin. (A) L. monocytogenes EGD-e/pNZ-P help -pHluorin was incubated at pH 6.5 in either LMB (solid bars) or a 1:1 mix of LMB with supernatant of L. lactis MG1363 (LMB:Sup; hatched bars) containing 5 μg/ml nisin. CTAB (0.005%) was used as a positive control (+) and untreated bacteria in either LMB or LMB:Sup without bacteriocin served as negative control (-). (B) OD 600 of L. lactis MG1363 (black) or B1627 (blue) grown in GM17 medium. (C) Supernatants were collected for the cultures shown in (B) at the indicated time points during growth and incubated with L. monocytogenes EGD-e/pNZ-P help -pHluorin (1:1 mix of supernatant with sensor strain in LMB). Values in (A) and (C) are ratios of fluorescence intensities (emission at 510 nm) after excitation at 400 and 470 nm and are mean ± standard deviation of two (A) or four (C) independent experiments using different supernatants and different cultures of the sensor strain.

    Article Snippet: A pHluorin coding sequence, which was codon-optimized for L. monocytogenes ( pHluorinLmo ; Table ) was synthesized and purchased as 752 bp DNA fragment from a commercial service provider (Eurofins Genomics, Ebersberg, Germany) and cloned into pJET1.2 according to manufacturer’s instructions (Thermo Scientific, Waltham, MA, United States).

    Techniques: Activity Assay, Incubation, Positive Control, Negative Control, Fluorescence, Standard Deviation

    Assay to determine membrane-damaging activity of bacteriocins using the ratiometric fluorescence properties of L. monocytogenes EGD-e/pNZ-P help -pHluorin. (A) Calibration curves of the ratio of fluorescence intensities (emission at 510 nm) after excitation at 400 and 470 nm at different pH for crude extracts (red) and permeabilized bacteria of L. monocytogenes EGD-e/pNZ-P help -pHluorin (green). (B,C) L. monocytogenes EGD-e/pNZ-P help -pHluorin was incubated in LMB pH 6.5 containing the indicated concentrations of nisin A (B) or pediocin PA-1 (C) and intracellular pH (pH i ) was calculated based in the ratio of fluorescence intensities (emission at 510 nm) after excitation at 400 and 470 nm using the calibration curve in (A) . CTAB (0.005%) was used as a positive control and untreated bacteria in LMB pH 6.5 without bacteriocin served as negative control (–). Values are mean ± standard deviation of three independent experiments using different cultures of the sensor strain. Statistical analysis was performed by ANOVA with Dunnett’s post-test to calculate p -values adjusted for multiple comparisons with untreated bacteria (–) set as control condition ( ∗ p

    Journal: Frontiers in Microbiology

    Article Title: Intracellular pHluorin as Sensor for Easy Assessment of Bacteriocin-Induced Membrane-Damage in Listeria monocytogenes

    doi: 10.3389/fmicb.2018.03038

    Figure Lengend Snippet: Assay to determine membrane-damaging activity of bacteriocins using the ratiometric fluorescence properties of L. monocytogenes EGD-e/pNZ-P help -pHluorin. (A) Calibration curves of the ratio of fluorescence intensities (emission at 510 nm) after excitation at 400 and 470 nm at different pH for crude extracts (red) and permeabilized bacteria of L. monocytogenes EGD-e/pNZ-P help -pHluorin (green). (B,C) L. monocytogenes EGD-e/pNZ-P help -pHluorin was incubated in LMB pH 6.5 containing the indicated concentrations of nisin A (B) or pediocin PA-1 (C) and intracellular pH (pH i ) was calculated based in the ratio of fluorescence intensities (emission at 510 nm) after excitation at 400 and 470 nm using the calibration curve in (A) . CTAB (0.005%) was used as a positive control and untreated bacteria in LMB pH 6.5 without bacteriocin served as negative control (–). Values are mean ± standard deviation of three independent experiments using different cultures of the sensor strain. Statistical analysis was performed by ANOVA with Dunnett’s post-test to calculate p -values adjusted for multiple comparisons with untreated bacteria (–) set as control condition ( ∗ p

    Article Snippet: A pHluorin coding sequence, which was codon-optimized for L. monocytogenes ( pHluorinLmo ; Table ) was synthesized and purchased as 752 bp DNA fragment from a commercial service provider (Eurofins Genomics, Ebersberg, Germany) and cloned into pJET1.2 according to manufacturer’s instructions (Thermo Scientific, Waltham, MA, United States).

    Techniques: Activity Assay, Fluorescence, Incubation, Positive Control, Negative Control, Standard Deviation

    Generation of the sensor strain L. monocytogenes EGD-e/pNZ-P help -pHluorin expressing ratiometric pHluorin. (A) Plasmid map of pNZ-P help -pHluorin for high level, constitutive expression of pHluorin encoded by the codon-optimized pHluorinLmo gene. (B) L. monocytogenes EGD-e/pNZ-P help -pHluorin (left) and L. monocytogenes EGD-e/pNZ44 (right) on BHI agar plate containing 15 μg/ml chloramphenicol. Expression of pHluorin was visualized with excitation at 455–485 nm and emission at 510–555 nm.

    Journal: Frontiers in Microbiology

    Article Title: Intracellular pHluorin as Sensor for Easy Assessment of Bacteriocin-Induced Membrane-Damage in Listeria monocytogenes

    doi: 10.3389/fmicb.2018.03038

    Figure Lengend Snippet: Generation of the sensor strain L. monocytogenes EGD-e/pNZ-P help -pHluorin expressing ratiometric pHluorin. (A) Plasmid map of pNZ-P help -pHluorin for high level, constitutive expression of pHluorin encoded by the codon-optimized pHluorinLmo gene. (B) L. monocytogenes EGD-e/pNZ-P help -pHluorin (left) and L. monocytogenes EGD-e/pNZ44 (right) on BHI agar plate containing 15 μg/ml chloramphenicol. Expression of pHluorin was visualized with excitation at 455–485 nm and emission at 510–555 nm.

    Article Snippet: A pHluorin coding sequence, which was codon-optimized for L. monocytogenes ( pHluorinLmo ; Table ) was synthesized and purchased as 752 bp DNA fragment from a commercial service provider (Eurofins Genomics, Ebersberg, Germany) and cloned into pJET1.2 according to manufacturer’s instructions (Thermo Scientific, Waltham, MA, United States).

    Techniques: Expressing, Plasmid Preparation

    Mean lag-phase duration (h), maximum growth rate (ΔOD 600 /h), and maximum OD 600 values for L. monocytogenes isolates possessing LGI1 ( n = 9) and isolates without LGI1 ( n = 8) grown in the presence of sublethal concentrations of E-San (1.6 μl/ml)

    Journal: Applied and Environmental Microbiology

    Article Title: Tolerance of Listeria monocytogenes to Quaternary Ammonium Sanitizers Is Mediated by a Novel Efflux Pump Encoded by emrE

    doi: 10.1128/AEM.03741-15

    Figure Lengend Snippet: Mean lag-phase duration (h), maximum growth rate (ΔOD 600 /h), and maximum OD 600 values for L. monocytogenes isolates possessing LGI1 ( n = 9) and isolates without LGI1 ( n = 8) grown in the presence of sublethal concentrations of E-San (1.6 μl/ml)

    Article Snippet: To assess the growth and survival of the isolates at different salt concentrations, L. monocytogenes 08-5578 and its LGI1 mutant derivative strains were exposed to NaCl solutions (5 to 20% [wt/vol]; Fisher Scientific) according to the protocol described by Ells and Truelstrup Hansen , with slight modifications.

    Techniques:

    Growth of L. monocytogenes 08-5578 (WT) and its Δ emrE Lm mutant in the presence of E-San at 0.8 μl/ml (A) and 1.6 μl/ml (B) or benzalkonium chloride (BAC) at 1 μg/ml (C) and 2 μg/ml (D) at 30°C, with (white

    Journal: Applied and Environmental Microbiology

    Article Title: Tolerance of Listeria monocytogenes to Quaternary Ammonium Sanitizers Is Mediated by a Novel Efflux Pump Encoded by emrE

    doi: 10.1128/AEM.03741-15

    Figure Lengend Snippet: Growth of L. monocytogenes 08-5578 (WT) and its Δ emrE Lm mutant in the presence of E-San at 0.8 μl/ml (A) and 1.6 μl/ml (B) or benzalkonium chloride (BAC) at 1 μg/ml (C) and 2 μg/ml (D) at 30°C, with (white

    Article Snippet: To assess the growth and survival of the isolates at different salt concentrations, L. monocytogenes 08-5578 and its LGI1 mutant derivative strains were exposed to NaCl solutions (5 to 20% [wt/vol]; Fisher Scientific) according to the protocol described by Ells and Truelstrup Hansen , with slight modifications.

    Techniques: Mutagenesis, BAC Assay

    Adhesion and invasion efficiencies (% bacteria recovered relative to the initial inoculum, normalized to the 08-5578 strain) of the L. monocytogenes WT strain (08-5578) and its isogenic mutant possessing a deletion in the sel1 gene, located on LGI1, compared

    Journal: Applied and Environmental Microbiology

    Article Title: Tolerance of Listeria monocytogenes to Quaternary Ammonium Sanitizers Is Mediated by a Novel Efflux Pump Encoded by emrE

    doi: 10.1128/AEM.03741-15

    Figure Lengend Snippet: Adhesion and invasion efficiencies (% bacteria recovered relative to the initial inoculum, normalized to the 08-5578 strain) of the L. monocytogenes WT strain (08-5578) and its isogenic mutant possessing a deletion in the sel1 gene, located on LGI1, compared

    Article Snippet: To assess the growth and survival of the isolates at different salt concentrations, L. monocytogenes 08-5578 and its LGI1 mutant derivative strains were exposed to NaCl solutions (5 to 20% [wt/vol]; Fisher Scientific) according to the protocol described by Ells and Truelstrup Hansen , with slight modifications.

    Techniques: Mutagenesis

    Growth of the L. monocytogenes 08-5578 WT strain (black squares) and its Δ emrE Lm (white squares), Δ lmo1851 (white triangles), and Δ sel1 (white circles) mutants in TSB with different concentrations of the E-San (A and B) and benzalkonium

    Journal: Applied and Environmental Microbiology

    Article Title: Tolerance of Listeria monocytogenes to Quaternary Ammonium Sanitizers Is Mediated by a Novel Efflux Pump Encoded by emrE

    doi: 10.1128/AEM.03741-15

    Figure Lengend Snippet: Growth of the L. monocytogenes 08-5578 WT strain (black squares) and its Δ emrE Lm (white squares), Δ lmo1851 (white triangles), and Δ sel1 (white circles) mutants in TSB with different concentrations of the E-San (A and B) and benzalkonium

    Article Snippet: To assess the growth and survival of the isolates at different salt concentrations, L. monocytogenes 08-5578 and its LGI1 mutant derivative strains were exposed to NaCl solutions (5 to 20% [wt/vol]; Fisher Scientific) according to the protocol described by Ells and Truelstrup Hansen , with slight modifications.

    Techniques:

    Expression profile of selected LGI1 genes when L. monocytogenes 08-5578 was grown in BHI supplemented with 5 μg/ml benzalkonium chloride at 37°C for 18 h.

    Journal: Applied and Environmental Microbiology

    Article Title: Tolerance of Listeria monocytogenes to Quaternary Ammonium Sanitizers Is Mediated by a Novel Efflux Pump Encoded by emrE

    doi: 10.1128/AEM.03741-15

    Figure Lengend Snippet: Expression profile of selected LGI1 genes when L. monocytogenes 08-5578 was grown in BHI supplemented with 5 μg/ml benzalkonium chloride at 37°C for 18 h.

    Article Snippet: To assess the growth and survival of the isolates at different salt concentrations, L. monocytogenes 08-5578 and its LGI1 mutant derivative strains were exposed to NaCl solutions (5 to 20% [wt/vol]; Fisher Scientific) according to the protocol described by Ells and Truelstrup Hansen , with slight modifications.

    Techniques: Expressing

    (A) 5′-Nuclease PCR analysis of serial 10-fold dilutions of L. monocytogenes DNA. C T values are plotted against the calculated CFU (i.e., 10-fold dilutions of the bacterial DNA from 3.125 × 10 6 CFU/μl). The straight line, which is calculated by linear regression ( y = −3.56 x [CFU] + 40.7) shows a square regression coefficient of R 2 = 0.993. The standard deviations based on three PCR reactions are indicated. (B) 5′-Nuclease PCR analysis of serial 10-fold dilutions of L. monocytogenes cells. C T values plotted against the number of CFU of L. monocytogenes . Template DNA was extracted from samples of cells containing serial 10-fold dilutions of approximately (5.0 ± 0.3) × 10 7 CFU of L. monocytogenes . The straight line, which is calculated by linear regression ( y = −4.12 x [CFU] + 45.5) shows a square regression coefficient of R 2 = 0.995. The standard deviations based on three PCR reactions are indicated.

    Journal: Applied and Environmental Microbiology

    Article Title: Application of 5?-Nuclease PCR for Quantitative Detection of Listeria monocytogenes in Pure Cultures, Water, Skim Milk, and Unpasteurized Whole Milk

    doi:

    Figure Lengend Snippet: (A) 5′-Nuclease PCR analysis of serial 10-fold dilutions of L. monocytogenes DNA. C T values are plotted against the calculated CFU (i.e., 10-fold dilutions of the bacterial DNA from 3.125 × 10 6 CFU/μl). The straight line, which is calculated by linear regression ( y = −3.56 x [CFU] + 40.7) shows a square regression coefficient of R 2 = 0.993. The standard deviations based on three PCR reactions are indicated. (B) 5′-Nuclease PCR analysis of serial 10-fold dilutions of L. monocytogenes cells. C T values plotted against the number of CFU of L. monocytogenes . Template DNA was extracted from samples of cells containing serial 10-fold dilutions of approximately (5.0 ± 0.3) × 10 7 CFU of L. monocytogenes . The straight line, which is calculated by linear regression ( y = −4.12 x [CFU] + 45.5) shows a square regression coefficient of R 2 = 0.995. The standard deviations based on three PCR reactions are indicated.

    Article Snippet: A total of 65 isolates of L. monocytogenes including the L. monocytogenes type strain, DSMZ 20600T (Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH, Braunschweig, Germany), were used to control the specificity of the primers and the probe.

    Techniques: Polymerase Chain Reaction

    Effect of pH i on actin polymerization-driven motility of L. monocytogenes . A , schematic of NHE1 and the point mutation that blocks ion translocation of NHE1 while maintaining its ability to interact with adaptor proteins through its long cytoplasmic tail

    Journal: The Journal of Biological Chemistry

    Article Title: Electrostatics Control Actin Filament Nucleation and Elongation Kinetics *

    doi: 10.1074/jbc.M113.456327

    Figure Lengend Snippet: Effect of pH i on actin polymerization-driven motility of L. monocytogenes . A , schematic of NHE1 and the point mutation that blocks ion translocation of NHE1 while maintaining its ability to interact with adaptor proteins through its long cytoplasmic tail

    Article Snippet: Before infection with L. monocytogenes , cells were plated in MatTek dishes containing an inserted coverslip, maintained in growth medium for 24–48 h, then washed three times with PBS, and incubated with DMEM containing 10% FBS without antibiotics.

    Techniques: Mutagenesis, Translocation Assay

    Increased bacterial loads after infection of TLR2 −/− mice with L. monocytogenes . TLR2 +/+ (black bars), TLR2 −/− (white bars), and MyD88 −/− (hatched bars) mice were infected with 2 × 10 5 CFU of Listeria i.v. and sacrificed at day 2 postinfection, and the total numbers of CFU per liver (A) and spleen (B) were determined. Results are from one representative experiment of two independent experiments. Results are expressed as means ± SD. *, P

    Journal: Infection and Immunity

    Article Title: Toll-Like Receptor 2 Is Required for Optimal Control of Listeria monocytogenes Infection

    doi: 10.1128/IAI.72.4.2131-2139.2004

    Figure Lengend Snippet: Increased bacterial loads after infection of TLR2 −/− mice with L. monocytogenes . TLR2 +/+ (black bars), TLR2 −/− (white bars), and MyD88 −/− (hatched bars) mice were infected with 2 × 10 5 CFU of Listeria i.v. and sacrificed at day 2 postinfection, and the total numbers of CFU per liver (A) and spleen (B) were determined. Results are from one representative experiment of two independent experiments. Results are expressed as means ± SD. *, P

    Article Snippet: L. monocytogenes (L028 strain from P. Cossart, Pasteur Institute, Paris, France) was cultured in Trypticase soy broth (soybean casein digest medium; Biovalley), divided into aliquots, and stored in 30% glycerol at −80°C at a concentration of 5 × 109 CFU/ml.

    Techniques: Infection, Mouse Assay

    Reduced CD40 and CD86 expression in TLR2- and MyD88-deficient macrophages and DCs infected with L. monocytogenes in vitro. BMDM from TLR2 +/+ , TLR2 −/− , and MyD88 −/− mice were stimulated for 24 h with Listeria (2×) (A), HKLM (200×) (B), BLP (0.5 μg/ml) (C), and LPS (100 ng/ml) (D). BMDM were labeled with an anti-CD11b antibody ( > 97% CD11b + ) and analyzed by fluorescence-activated cell sorting for CD40 expression. Unstimulated controls showed essentially no CD40 expression, similar to the isotype controls (dotted lines). Results are from one representative experiment (a pool of two mice) of three independent experiments. The levels of CD40 (E) and CD86 (F) expression by CD11b + BMDM and CD11c + BMDC from TLR2 +/+ , TLR2 −/− , and MyD88 −/− mice stimulated for 24 h by Listeria (2×) were also compared. The results, expressed as geometric mean fluorescence intensities relative to those of wild-type controls, were calculated from three independent experiments and are expressed as means ± SD. *, P

    Journal: Infection and Immunity

    Article Title: Toll-Like Receptor 2 Is Required for Optimal Control of Listeria monocytogenes Infection

    doi: 10.1128/IAI.72.4.2131-2139.2004

    Figure Lengend Snippet: Reduced CD40 and CD86 expression in TLR2- and MyD88-deficient macrophages and DCs infected with L. monocytogenes in vitro. BMDM from TLR2 +/+ , TLR2 −/− , and MyD88 −/− mice were stimulated for 24 h with Listeria (2×) (A), HKLM (200×) (B), BLP (0.5 μg/ml) (C), and LPS (100 ng/ml) (D). BMDM were labeled with an anti-CD11b antibody ( > 97% CD11b + ) and analyzed by fluorescence-activated cell sorting for CD40 expression. Unstimulated controls showed essentially no CD40 expression, similar to the isotype controls (dotted lines). Results are from one representative experiment (a pool of two mice) of three independent experiments. The levels of CD40 (E) and CD86 (F) expression by CD11b + BMDM and CD11c + BMDC from TLR2 +/+ , TLR2 −/− , and MyD88 −/− mice stimulated for 24 h by Listeria (2×) were also compared. The results, expressed as geometric mean fluorescence intensities relative to those of wild-type controls, were calculated from three independent experiments and are expressed as means ± SD. *, P

    Article Snippet: L. monocytogenes (L028 strain from P. Cossart, Pasteur Institute, Paris, France) was cultured in Trypticase soy broth (soybean casein digest medium; Biovalley), divided into aliquots, and stored in 30% glycerol at −80°C at a concentration of 5 × 109 CFU/ml.

    Techniques: Expressing, Infection, In Vitro, Mouse Assay, Labeling, Fluorescence, FACS

    Requirement of TLR2 for IFN-γ induction in early L. monocytogenes infection. TLR2 +/+ (white circles) and TLR2 −/− (black circles) mice were inoculated i.v. with 2 × 10 5 CFU of Listeria . Blood was taken at 24 h postinfection, and IFN-γ levels in serum were measured by ELISA. Results are from one representative experiment of two independent experiments and are expressed as means ± SD. *, P

    Journal: Infection and Immunity

    Article Title: Toll-Like Receptor 2 Is Required for Optimal Control of Listeria monocytogenes Infection

    doi: 10.1128/IAI.72.4.2131-2139.2004

    Figure Lengend Snippet: Requirement of TLR2 for IFN-γ induction in early L. monocytogenes infection. TLR2 +/+ (white circles) and TLR2 −/− (black circles) mice were inoculated i.v. with 2 × 10 5 CFU of Listeria . Blood was taken at 24 h postinfection, and IFN-γ levels in serum were measured by ELISA. Results are from one representative experiment of two independent experiments and are expressed as means ± SD. *, P

    Article Snippet: L. monocytogenes (L028 strain from P. Cossart, Pasteur Institute, Paris, France) was cultured in Trypticase soy broth (soybean casein digest medium; Biovalley), divided into aliquots, and stored in 30% glycerol at −80°C at a concentration of 5 × 109 CFU/ml.

    Techniques: Infection, Mouse Assay, Enzyme-linked Immunosorbent Assay

    Increased susceptibility of TLR2 −/− mice to L. monocytogenes . TLR2 +/+ (triangles) and TLR2 −/− (squares) mice were inoculated i.v. with 3 × 10 5 CFU/mouse (A) and 1 × 10 5 CFU/mouse (B) and were monitored for survival. For panel A, P = 0.03, and for panel B, P = 0.022 by the log rank test. (A) Results are from 11 TLR2 +/+ mice and 9 TLR2 −/− mice. (B) Results are from 14 mice per group. The results shown are the combination of two identical experiments.

    Journal: Infection and Immunity

    Article Title: Toll-Like Receptor 2 Is Required for Optimal Control of Listeria monocytogenes Infection

    doi: 10.1128/IAI.72.4.2131-2139.2004

    Figure Lengend Snippet: Increased susceptibility of TLR2 −/− mice to L. monocytogenes . TLR2 +/+ (triangles) and TLR2 −/− (squares) mice were inoculated i.v. with 3 × 10 5 CFU/mouse (A) and 1 × 10 5 CFU/mouse (B) and were monitored for survival. For panel A, P = 0.03, and for panel B, P = 0.022 by the log rank test. (A) Results are from 11 TLR2 +/+ mice and 9 TLR2 −/− mice. (B) Results are from 14 mice per group. The results shown are the combination of two identical experiments.

    Article Snippet: L. monocytogenes (L028 strain from P. Cossart, Pasteur Institute, Paris, France) was cultured in Trypticase soy broth (soybean casein digest medium; Biovalley), divided into aliquots, and stored in 30% glycerol at −80°C at a concentration of 5 × 109 CFU/ml.

    Techniques: Mouse Assay

    Immune cell recruitment to microabscesses after L. monocytogenes infection. Images of the immunohistochemistry of TLR2 +/+ (A, C, E, and G) and TLR2 −/− (B, D, F, and H) livers 2 days after i.v. infection with 2 × 10 5 CFU/mouse are shown. Immunolabeling with GR1 (A and B), F4/80 (C and D), CD11b (E and F), and iNOS (G and H) is shown in brown (magnification, ×200).

    Journal: Infection and Immunity

    Article Title: Toll-Like Receptor 2 Is Required for Optimal Control of Listeria monocytogenes Infection

    doi: 10.1128/IAI.72.4.2131-2139.2004

    Figure Lengend Snippet: Immune cell recruitment to microabscesses after L. monocytogenes infection. Images of the immunohistochemistry of TLR2 +/+ (A, C, E, and G) and TLR2 −/− (B, D, F, and H) livers 2 days after i.v. infection with 2 × 10 5 CFU/mouse are shown. Immunolabeling with GR1 (A and B), F4/80 (C and D), CD11b (E and F), and iNOS (G and H) is shown in brown (magnification, ×200).

    Article Snippet: L. monocytogenes (L028 strain from P. Cossart, Pasteur Institute, Paris, France) was cultured in Trypticase soy broth (soybean casein digest medium; Biovalley), divided into aliquots, and stored in 30% glycerol at −80°C at a concentration of 5 × 109 CFU/ml.

    Techniques: Infection, Immunohistochemistry, Immunolabeling

    Growth of L. monocytogenes on cooked, modified-atmosphere, hot-packed poultry injected with regular brine (open symbols) or test brine containing sodium lactate and ALTA 2341 (solid symbols) and stored at 3.5°C (▴), 6.5°C (•), or 10°C (■). Counts below 1,000 CFU/piece were determined by MPN.

    Journal: Applied and Environmental Microbiology

    Article Title: Growth of Listeria monocytogenes and Yersinia enterocolitica on Cooked Modified-Atmosphere-Packaged Poultry in the Presence and Absence of a Naturally Occurring Microbiota

    doi:

    Figure Lengend Snippet: Growth of L. monocytogenes on cooked, modified-atmosphere, hot-packed poultry injected with regular brine (open symbols) or test brine containing sodium lactate and ALTA 2341 (solid symbols) and stored at 3.5°C (▴), 6.5°C (•), or 10°C (■). Counts below 1,000 CFU/piece were determined by MPN.

    Article Snippet: The identification of selected colonies enumerated as either Y. enterocolitica or L. monocytogenes was confirmed by Vitek Jr. (bioMérieux Vitek, Inc., Hazelwood, Mo.).

    Techniques: Modification, Injection

    Restriction-modification system genes in the L. monocytogenes serotype 4b genome. Annotation and visualization of the region were done using GAMOLA and ARTEMIS as described in Materials and Methods. Putative open reading frames designated 5-methyl cytosine restriction, methylase, and putative Sau3AI correspond to 85R, 85M, and 85S, respectively, described in the text. Black bars indicate probes used for Southern blot-based detection of 85R, 85M, and 85S. Arrows indicate direction of transcription. The deduced G+C contents of the genes are shown at the top.

    Journal: Applied and Environmental Microbiology

    Article Title: Epidemic Clone I-Specific Genetic Markers in Strains of Listeria monocytogenes Serotype 4b from Foods

    doi: 10.1128/AEM.70.7.4158-4164.2004

    Figure Lengend Snippet: Restriction-modification system genes in the L. monocytogenes serotype 4b genome. Annotation and visualization of the region were done using GAMOLA and ARTEMIS as described in Materials and Methods. Putative open reading frames designated 5-methyl cytosine restriction, methylase, and putative Sau3AI correspond to 85R, 85M, and 85S, respectively, described in the text. Black bars indicate probes used for Southern blot-based detection of 85R, 85M, and 85S. Arrows indicate direction of transcription. The deduced G+C contents of the genes are shown at the top.

    Article Snippet: Some primers were designed based on preliminary sequence data of L. monocytogenes ) and were purchased from Bio-synthesis, Inc. (Lewisville, Tex.) or from QIAGEN.

    Techniques: Modification, Southern Blot

    The AgDI pathway and organization of the gene cluster in L. monocytogenes . A , the AgDI pathway. Agmatine is first hydrolyzed to form N -carbamoylputrescine by AgDI. N -Carbamoylputrescine is then converted into putrescine and carbamoylphosphate by putrescine

    Journal: The Journal of Biological Chemistry

    Article Title: Listeria monocytogenes aguA1, but Not aguA2, Encodes a Functional Agmatine Deiminase

    doi: 10.1074/jbc.M113.477380

    Figure Lengend Snippet: The AgDI pathway and organization of the gene cluster in L. monocytogenes . A , the AgDI pathway. Agmatine is first hydrolyzed to form N -carbamoylputrescine by AgDI. N -Carbamoylputrescine is then converted into putrescine and carbamoylphosphate by putrescine

    Article Snippet: The amino acid sequences of AguA1 and AguA2 of L. monocytogenes and its homologs in other microbial species were obtained from the National Centre for Biotechnology Information database.

    Techniques:

    Responses or roles of aguA1 and aguA2 under acidic stresses. A , relative quantification of aguA1 and aguA2 mRNA levels in L. monocytogenes 10403S exposed to BHI medium at pH 5.0 and 7.0. Values are expressed as means ± S.D. B , survival of L. monocytogenes

    Journal: The Journal of Biological Chemistry

    Article Title: Listeria monocytogenes aguA1, but Not aguA2, Encodes a Functional Agmatine Deiminase

    doi: 10.1074/jbc.M113.477380

    Figure Lengend Snippet: Responses or roles of aguA1 and aguA2 under acidic stresses. A , relative quantification of aguA1 and aguA2 mRNA levels in L. monocytogenes 10403S exposed to BHI medium at pH 5.0 and 7.0. Values are expressed as means ± S.D. B , survival of L. monocytogenes

    Article Snippet: The amino acid sequences of AguA1 and AguA2 of L. monocytogenes and its homologs in other microbial species were obtained from the National Centre for Biotechnology Information database.

    Techniques:

    Comparison of ready-to-eat (RTE) sample contigs against the Listeria monocytogenes EGD-e chromosome, complete genome (NC_003210.1), displayed in the inner black circle.

    Journal: Genes

    Article Title: Genomic Diversity of Common Sequence Types of Listeria monocytogenes Isolated from Ready-to-Eat Products of Animal Origin in South Africa

    doi: 10.3390/genes10121007

    Figure Lengend Snippet: Comparison of ready-to-eat (RTE) sample contigs against the Listeria monocytogenes EGD-e chromosome, complete genome (NC_003210.1), displayed in the inner black circle.

    Article Snippet: Data Availability The six genome sequences of L. monocytogenes isolates were deposited at the National Centre for Biotechnology Information (NCBI)/GenBank under the accession numbers from SAMN12360665 to SAMN12360670 (BioProject No. PRJNA556582).

    Techniques:

    Listeria monocytogenes CDS average nucleotide identities using Gower distance metric.

    Journal: Genes

    Article Title: Genomic Diversity of Common Sequence Types of Listeria monocytogenes Isolated from Ready-to-Eat Products of Animal Origin in South Africa

    doi: 10.3390/genes10121007

    Figure Lengend Snippet: Listeria monocytogenes CDS average nucleotide identities using Gower distance metric.

    Article Snippet: Data Availability The six genome sequences of L. monocytogenes isolates were deposited at the National Centre for Biotechnology Information (NCBI)/GenBank under the accession numbers from SAMN12360665 to SAMN12360670 (BioProject No. PRJNA556582).

    Techniques:

    L. monocytogenes (LM) is trapped in late endosomes and acidified lysosomes in EVT. ( A ) Percent co-localization of early endosomal marker Rab5 and autophagosomal marker LC3 with LM. ( B ) Percent co-localization of late endosomal marker Rab7, lysosomal marker Lamp1 and acidotropic dye Lysotracker (LT) with LM. p.i. = post-inoculation. Each data point is an average of 3 independent experiments. Bars represent SEM. ( C–E ) Representative images of GFP-expressing LM (green) co-localizing with markers Rab7, Lamp1, and LT (red). The color-merged images also show DAPI counterstain (white). Scale bars are 10 µm.

    Journal: PLoS Pathogens

    Article Title: Invasive Extravillous Trophoblasts Restrict Intracellular Growth and Spread of Listeria monocytogenes

    doi: 10.1371/journal.ppat.1002005

    Figure Lengend Snippet: L. monocytogenes (LM) is trapped in late endosomes and acidified lysosomes in EVT. ( A ) Percent co-localization of early endosomal marker Rab5 and autophagosomal marker LC3 with LM. ( B ) Percent co-localization of late endosomal marker Rab7, lysosomal marker Lamp1 and acidotropic dye Lysotracker (LT) with LM. p.i. = post-inoculation. Each data point is an average of 3 independent experiments. Bars represent SEM. ( C–E ) Representative images of GFP-expressing LM (green) co-localizing with markers Rab7, Lamp1, and LT (red). The color-merged images also show DAPI counterstain (white). Scale bars are 10 µm.

    Article Snippet: The pactA -RFP strain (PL512) was constructed as follows: The ORF encoding TagRFP from Entacmaea quadricolor was codon optimized for expression in L. monocytogenes using Gene Designer software and the gene was synthesized de novo (DNA2.0, Menlo Park, CA).

    Techniques: Marker, Expressing

    L. monocytogenes (LM) in EVT is largely confined to vacuoles. ( A,B ) Representative transmission electron micrographs from 2 ( A ) and 5 ( B ) hours post-inoculation (p.i.). Bacteria are marked with asterisks. Scale bars are 2 and 1 µm respectively. ( C ) Quantification of subcellular localization of bacteria in EVT at 2 and 5 hours p.i. ( D ) Quantification of intact versus degraded bacteria in vacuoles in EVT at 2 and 5 hours p.i. ( E ) Close-up image at 5 hours p.i. shows a single membrane vacuole surrounding intact bacterium. Scale bar is 100 nm. ( F ) Close-up image of degraded bacterium in a vacuole in EVT at 5 hours p.i. Scale bar is 2 µm.

    Journal: PLoS Pathogens

    Article Title: Invasive Extravillous Trophoblasts Restrict Intracellular Growth and Spread of Listeria monocytogenes

    doi: 10.1371/journal.ppat.1002005

    Figure Lengend Snippet: L. monocytogenes (LM) in EVT is largely confined to vacuoles. ( A,B ) Representative transmission electron micrographs from 2 ( A ) and 5 ( B ) hours post-inoculation (p.i.). Bacteria are marked with asterisks. Scale bars are 2 and 1 µm respectively. ( C ) Quantification of subcellular localization of bacteria in EVT at 2 and 5 hours p.i. ( D ) Quantification of intact versus degraded bacteria in vacuoles in EVT at 2 and 5 hours p.i. ( E ) Close-up image at 5 hours p.i. shows a single membrane vacuole surrounding intact bacterium. Scale bar is 100 nm. ( F ) Close-up image of degraded bacterium in a vacuole in EVT at 5 hours p.i. Scale bar is 2 µm.

    Article Snippet: The pactA -RFP strain (PL512) was constructed as follows: The ORF encoding TagRFP from Entacmaea quadricolor was codon optimized for expression in L. monocytogenes using Gene Designer software and the gene was synthesized de novo (DNA2.0, Menlo Park, CA).

    Techniques: Transmission Assay

    L. monocytogenes (LM) is trapped in late endosomes and lysosomes in EVT of placental explants. ( A ) Representative image of placental explant 8 hours post-inoculation (p.i.) with p actA -RFP LM counterstained with anti-LM antibody (green). DAPI counterstain (white). Scale bar is 50 µm. ( B ) Close-up of boxed area in panel A. Scale bar is 10 µm. ( C ) Vacuolar escape measured by percent RFP expression of p actA -RFP strain counterstained with anti-LM antibody (green). Percent co-localization of LM with late endosomal marker Rab7 and lysosomal marker Lamp1. Each data point is an average of 3 independent experiments. Bars represent SEM.

    Journal: PLoS Pathogens

    Article Title: Invasive Extravillous Trophoblasts Restrict Intracellular Growth and Spread of Listeria monocytogenes

    doi: 10.1371/journal.ppat.1002005

    Figure Lengend Snippet: L. monocytogenes (LM) is trapped in late endosomes and lysosomes in EVT of placental explants. ( A ) Representative image of placental explant 8 hours post-inoculation (p.i.) with p actA -RFP LM counterstained with anti-LM antibody (green). DAPI counterstain (white). Scale bar is 50 µm. ( B ) Close-up of boxed area in panel A. Scale bar is 10 µm. ( C ) Vacuolar escape measured by percent RFP expression of p actA -RFP strain counterstained with anti-LM antibody (green). Percent co-localization of LM with late endosomal marker Rab7 and lysosomal marker Lamp1. Each data point is an average of 3 independent experiments. Bars represent SEM.

    Article Snippet: The pactA -RFP strain (PL512) was constructed as follows: The ORF encoding TagRFP from Entacmaea quadricolor was codon optimized for expression in L. monocytogenes using Gene Designer software and the gene was synthesized de novo (DNA2.0, Menlo Park, CA).

    Techniques: Expressing, Marker

    L. monocytogenes (LM) vacuolar escape rates vary across subpopulations of trophoblasts. ( A ) Representative image of placental explant 24 hours post-inoculation (p.i.) with p actA -RFP LM counterstained with anti-LM (green). DAPI counterstain (white). STR = stroma. ( B,C ) Close-up examples of Invasive Border EVT ( B ), Parastromal trophoblasts and middle EVT ( C ). ( D ) Rotated image of placental explant from panel A with superimposed surface plot representing areas of highest RFP expression in relation to total bacterial green signal. Highest peaks are in the parastromal trophoblast region, where vacuolar escape rates are highest. ( E ) Percent RFP-expressing bacteria in each trophoblast subpopulation were enumerated at 24 and 48 hours p.i. in villi where all 3 subpopulations were infected. RFP expression was normalized to the 24-hour time point in invasive border EVT from the same placenta. Results from 3 independent experiments were expressed as average fold-change in comparison to the escape rate in invasive border EVT at 24 hours. Bars are SEM. Asterisk represents significant difference (p = 0.02).

    Journal: PLoS Pathogens

    Article Title: Invasive Extravillous Trophoblasts Restrict Intracellular Growth and Spread of Listeria monocytogenes

    doi: 10.1371/journal.ppat.1002005

    Figure Lengend Snippet: L. monocytogenes (LM) vacuolar escape rates vary across subpopulations of trophoblasts. ( A ) Representative image of placental explant 24 hours post-inoculation (p.i.) with p actA -RFP LM counterstained with anti-LM (green). DAPI counterstain (white). STR = stroma. ( B,C ) Close-up examples of Invasive Border EVT ( B ), Parastromal trophoblasts and middle EVT ( C ). ( D ) Rotated image of placental explant from panel A with superimposed surface plot representing areas of highest RFP expression in relation to total bacterial green signal. Highest peaks are in the parastromal trophoblast region, where vacuolar escape rates are highest. ( E ) Percent RFP-expressing bacteria in each trophoblast subpopulation were enumerated at 24 and 48 hours p.i. in villi where all 3 subpopulations were infected. RFP expression was normalized to the 24-hour time point in invasive border EVT from the same placenta. Results from 3 independent experiments were expressed as average fold-change in comparison to the escape rate in invasive border EVT at 24 hours. Bars are SEM. Asterisk represents significant difference (p = 0.02).

    Article Snippet: The pactA -RFP strain (PL512) was constructed as follows: The ORF encoding TagRFP from Entacmaea quadricolor was codon optimized for expression in L. monocytogenes using Gene Designer software and the gene was synthesized de novo (DNA2.0, Menlo Park, CA).

    Techniques: Expressing, Infection

    Vacuolar escape of L. monocytogenes (LM) is strongly impaired in EVT. BeWo cells and EVT were infected with p actA -RFP LM and counterstained with anti-LM antibody (green). ( A,B ) Representative images of BeWo ( A ) and EVT ( B ) 8 hours post-inoculation (p.i.). DAPI counterstain (white). Scale bar is 10 µm. ( C ) Percent RFP-expressing bacteria were enumerated and represent bacteria that escaped from the primary vacuole. Actin nucleation by wild type LM in BeWo cells was measured by co-localization with phalloidin. Bacterial RFP expression and phalloidin-co-localization are expressed as a percentage of the total number of intracellular bacteria. Each data point is an average of 3 independent experiments. Bars represent SEM.

    Journal: PLoS Pathogens

    Article Title: Invasive Extravillous Trophoblasts Restrict Intracellular Growth and Spread of Listeria monocytogenes

    doi: 10.1371/journal.ppat.1002005

    Figure Lengend Snippet: Vacuolar escape of L. monocytogenes (LM) is strongly impaired in EVT. BeWo cells and EVT were infected with p actA -RFP LM and counterstained with anti-LM antibody (green). ( A,B ) Representative images of BeWo ( A ) and EVT ( B ) 8 hours post-inoculation (p.i.). DAPI counterstain (white). Scale bar is 10 µm. ( C ) Percent RFP-expressing bacteria were enumerated and represent bacteria that escaped from the primary vacuole. Actin nucleation by wild type LM in BeWo cells was measured by co-localization with phalloidin. Bacterial RFP expression and phalloidin-co-localization are expressed as a percentage of the total number of intracellular bacteria. Each data point is an average of 3 independent experiments. Bars represent SEM.

    Article Snippet: The pactA -RFP strain (PL512) was constructed as follows: The ORF encoding TagRFP from Entacmaea quadricolor was codon optimized for expression in L. monocytogenes using Gene Designer software and the gene was synthesized de novo (DNA2.0, Menlo Park, CA).

    Techniques: Infection, Expressing

    EVT restrict intracellular growth of L. monocytogenes . Intracellular growth curves of wild type L. monocytogenes in 3 cell types: primary placental fibroblasts (PF), choriocarcinoma cell line (BeWo), and primary extravillous trophoblasts (EVT). CFU/well were normalized to the 2-hour time point within each experiment. Each data point is an average of 3 independent experiments for PF and BeWo, and 10 independent experiments for EVT. Bars represent SEM.

    Journal: PLoS Pathogens

    Article Title: Invasive Extravillous Trophoblasts Restrict Intracellular Growth and Spread of Listeria monocytogenes

    doi: 10.1371/journal.ppat.1002005

    Figure Lengend Snippet: EVT restrict intracellular growth of L. monocytogenes . Intracellular growth curves of wild type L. monocytogenes in 3 cell types: primary placental fibroblasts (PF), choriocarcinoma cell line (BeWo), and primary extravillous trophoblasts (EVT). CFU/well were normalized to the 2-hour time point within each experiment. Each data point is an average of 3 independent experiments for PF and BeWo, and 10 independent experiments for EVT. Bars represent SEM.

    Article Snippet: The pactA -RFP strain (PL512) was constructed as follows: The ORF encoding TagRFP from Entacmaea quadricolor was codon optimized for expression in L. monocytogenes using Gene Designer software and the gene was synthesized de novo (DNA2.0, Menlo Park, CA).

    Techniques: