Structured Review

Difco sihumi members
<t>SIHUMI</t> mice colonized with both A. muciniphila and S. <t>Typhimurium</t> display an increased cecal macrophage infiltration. (A) Formalin fixed paraffin embedded cecum tissue was thin sectioned at 2 µm. Macrophages were stained by targeting the F4/80 receptor expressed on mouse macrophages using immunohistochemistry with specific antibodies. Brown color indicates positively stained macrophages. Magnification 400-fold. Bar = 100 µm. (B) Positively stained macrophages were enumerated along a stretch of 50 µm of lamina muscularis for both lamina propria and sub-mucosa (see materials and methods). SIHUMI mice colonized with both A. muciniphila and S. Typhimurium had the highest macrophage infiltration scores compared to the other groups (see Figure. 1). Data are expressed as median with range. n = 5 mice per group. *P
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1) Product Images from "Commensal Akkermansia muciniphila Exacerbates Gut Inflammation in Salmonella Typhimurium-Infected Gnotobiotic Mice"

Article Title: Commensal Akkermansia muciniphila Exacerbates Gut Inflammation in Salmonella Typhimurium-Infected Gnotobiotic Mice

Journal: PLoS ONE

doi: 10.1371/journal.pone.0074963

SIHUMI mice colonized with both A. muciniphila and S. Typhimurium display an increased cecal macrophage infiltration. (A) Formalin fixed paraffin embedded cecum tissue was thin sectioned at 2 µm. Macrophages were stained by targeting the F4/80 receptor expressed on mouse macrophages using immunohistochemistry with specific antibodies. Brown color indicates positively stained macrophages. Magnification 400-fold. Bar = 100 µm. (B) Positively stained macrophages were enumerated along a stretch of 50 µm of lamina muscularis for both lamina propria and sub-mucosa (see materials and methods). SIHUMI mice colonized with both A. muciniphila and S. Typhimurium had the highest macrophage infiltration scores compared to the other groups (see Figure. 1). Data are expressed as median with range. n = 5 mice per group. *P
Figure Legend Snippet: SIHUMI mice colonized with both A. muciniphila and S. Typhimurium display an increased cecal macrophage infiltration. (A) Formalin fixed paraffin embedded cecum tissue was thin sectioned at 2 µm. Macrophages were stained by targeting the F4/80 receptor expressed on mouse macrophages using immunohistochemistry with specific antibodies. Brown color indicates positively stained macrophages. Magnification 400-fold. Bar = 100 µm. (B) Positively stained macrophages were enumerated along a stretch of 50 µm of lamina muscularis for both lamina propria and sub-mucosa (see materials and methods). SIHUMI mice colonized with both A. muciniphila and S. Typhimurium had the highest macrophage infiltration scores compared to the other groups (see Figure. 1). Data are expressed as median with range. n = 5 mice per group. *P

Techniques Used: Mouse Assay, Formalin-fixed Paraffin-Embedded, Staining, Immunohistochemistry

Concomitant presence of A. muciniphila and S. Typhimurium results in increased histopathology scores in SIHUMI mice. (A) Gnotobiotic C3H mice containing 8 defined microbial species (SIHUMI) were subsequently inoculated with A. muciniphila or S. Typhimurium or consecutively with both organisms (see Figure 1 ). SIHUMI and SIHUMI-A mice had the lowest histopathology scores (≤4.0) with no signs of inflammation and were therefore taken as baseline (dotted line). Data are expressed as median with range. *P
Figure Legend Snippet: Concomitant presence of A. muciniphila and S. Typhimurium results in increased histopathology scores in SIHUMI mice. (A) Gnotobiotic C3H mice containing 8 defined microbial species (SIHUMI) were subsequently inoculated with A. muciniphila or S. Typhimurium or consecutively with both organisms (see Figure 1 ). SIHUMI and SIHUMI-A mice had the lowest histopathology scores (≤4.0) with no signs of inflammation and were therefore taken as baseline (dotted line). Data are expressed as median with range. *P

Techniques Used: Histopathology, Mouse Assay

SIHUMI mice colonized with both A. muciniphila and S. Typhimurium display enlarged mLN and elevated S. Typhimurium cell numbers. (A) Mesenteric lymph nodes (mLN) were obtained from four groups of gnotobiotic C3H mice. SIHUMI mice were subsequently inoculated with A. muciniphila or S. Typhimurium or consecutively with both organisms (see Figure 1 ). The mLN tissue was homogenized and DNA was isolated to quantify S. Typhimurium using quantitative PCR with primers targeting the ttr-region of S. Typhimurium. Absolute cell numbers were calculated based on calibration curves with known concentrations of S. Typhimurium. The mLN of SIHUMI-AS mice contained 10-fold higher cell numbers of S. Typhimurium compared to SIHUMI-S mice. Data are expressed as mean±standard error. n = 10 mice per group. *P
Figure Legend Snippet: SIHUMI mice colonized with both A. muciniphila and S. Typhimurium display enlarged mLN and elevated S. Typhimurium cell numbers. (A) Mesenteric lymph nodes (mLN) were obtained from four groups of gnotobiotic C3H mice. SIHUMI mice were subsequently inoculated with A. muciniphila or S. Typhimurium or consecutively with both organisms (see Figure 1 ). The mLN tissue was homogenized and DNA was isolated to quantify S. Typhimurium using quantitative PCR with primers targeting the ttr-region of S. Typhimurium. Absolute cell numbers were calculated based on calibration curves with known concentrations of S. Typhimurium. The mLN of SIHUMI-AS mice contained 10-fold higher cell numbers of S. Typhimurium compared to SIHUMI-S mice. Data are expressed as mean±standard error. n = 10 mice per group. *P

Techniques Used: Mouse Assay, Isolation, Real-time Polymerase Chain Reaction

SIHUMI mice colonized with both A. muciniphila and S. Typhimurium display reduced mucus sulphation. Formalin fixed thin sections (4 µm) of cecal tissue of mice belonging to either one of four groups: SIHUMI, SIHUMI-A, SIHUMI-S and SIHUMI-AS (see Figure. 1) were stained with high iron diamine (HID)/AB at pH-2.5 and subsequently analyzed. Brown color indicates sulphated mucins while blue color indicates sialylated mucins. SIHUMI-AS mice display few sulphated mucins compared to the other mouse groups. Magnification 400×. Bars indicate 100 µm.
Figure Legend Snippet: SIHUMI mice colonized with both A. muciniphila and S. Typhimurium display reduced mucus sulphation. Formalin fixed thin sections (4 µm) of cecal tissue of mice belonging to either one of four groups: SIHUMI, SIHUMI-A, SIHUMI-S and SIHUMI-AS (see Figure. 1) were stained with high iron diamine (HID)/AB at pH-2.5 and subsequently analyzed. Brown color indicates sulphated mucins while blue color indicates sialylated mucins. SIHUMI-AS mice display few sulphated mucins compared to the other mouse groups. Magnification 400×. Bars indicate 100 µm.

Techniques Used: Mouse Assay, Staining

Hypothetical Scheme. The presence of A. muciniphila, leads to the exacerbation of S. Typhimurium-induced intestinal inflammation. We propose that the presence of A. muciniphila causes changes in mucin composition and production, which in turn facilitates the invasion of S. Typhimurium into the host. Increased inflammatory status was characterized by increased pro-inflammatory cytokines, increased macrophage infiltration and invasion of the pathogen into the lymph nodes, reduced number of mucin-filled goblet cells in SIHUMI-AS mice (B) compared to SIHUMI-S mice (A). Our data suggests that in the presence of both A. muciniphila and S. Typhimurium, mucus sulphation is diminished and this may facilitate the access of S. Typhimurium to sialic acid in mucus. Sialic acid may serve as a substrate and adhesion site for S. Typhimurium in the gut [56] , [57] . Increased gene expression of IFN-γ and IP-10 indicate an increased NK-cell recruitment. mLN - mesenteric lymph nodes, NK- Natural killer cells. (↑ increased; ↓ decreased; grey dotted line: assumed processes including lectin-sialic acid binding [56] , M-cells for pathogen transit [43] , [58] , [59] ; black line: supported by data of the present study).
Figure Legend Snippet: Hypothetical Scheme. The presence of A. muciniphila, leads to the exacerbation of S. Typhimurium-induced intestinal inflammation. We propose that the presence of A. muciniphila causes changes in mucin composition and production, which in turn facilitates the invasion of S. Typhimurium into the host. Increased inflammatory status was characterized by increased pro-inflammatory cytokines, increased macrophage infiltration and invasion of the pathogen into the lymph nodes, reduced number of mucin-filled goblet cells in SIHUMI-AS mice (B) compared to SIHUMI-S mice (A). Our data suggests that in the presence of both A. muciniphila and S. Typhimurium, mucus sulphation is diminished and this may facilitate the access of S. Typhimurium to sialic acid in mucus. Sialic acid may serve as a substrate and adhesion site for S. Typhimurium in the gut [56] , [57] . Increased gene expression of IFN-γ and IP-10 indicate an increased NK-cell recruitment. mLN - mesenteric lymph nodes, NK- Natural killer cells. (↑ increased; ↓ decreased; grey dotted line: assumed processes including lectin-sialic acid binding [56] , M-cells for pathogen transit [43] , [58] , [59] ; black line: supported by data of the present study).

Techniques Used: Mouse Assay, Expressing, Binding Assay

Presence of A. muciniphila renders S. Typhimurium the dominant species in gnotobiotic SIHUMI mice. Cecal contents were collected from gnotobiotic C3H mice, differing in their microbial status: (A) Mice with a defined microbial community of eight bacterial species (SIHUMI), (B) SIHUMI mice additionally colonized with A. muciniphila (SIHUMI-A), (C) SIHUMI mice infected with S. Typhimurium (SIHUMI-S) and (D) SIHUMI mice colonized with A. muciniphila and 10 days later infected with S. Typhimurium (SIHUMI-AS) (see Figure 1 ). Total DNA was extracted and bacterial cell numbers were quantified by qPCR with primers targeting the HSP60 gene of the SIHUMI members, the 16S rRNA gene of A. muciniphila and the ttr-region of S. Typhimurium. Calculation of the cell numbers was based on DNA obtained from cell suspensions containing known cell numbers of the targeted bacterial species (see materials and methods). Presence of A. muciniphila in SIHUMI-AS mice is attributed to an increase in the proportion of S. Typhimurium cells at the expense of other community members showing reduced proportion of SIHUMI members. Ten animals per group were used. The bacterial cell numbers and P-values for the differences between the groups are provided in Table 1 .
Figure Legend Snippet: Presence of A. muciniphila renders S. Typhimurium the dominant species in gnotobiotic SIHUMI mice. Cecal contents were collected from gnotobiotic C3H mice, differing in their microbial status: (A) Mice with a defined microbial community of eight bacterial species (SIHUMI), (B) SIHUMI mice additionally colonized with A. muciniphila (SIHUMI-A), (C) SIHUMI mice infected with S. Typhimurium (SIHUMI-S) and (D) SIHUMI mice colonized with A. muciniphila and 10 days later infected with S. Typhimurium (SIHUMI-AS) (see Figure 1 ). Total DNA was extracted and bacterial cell numbers were quantified by qPCR with primers targeting the HSP60 gene of the SIHUMI members, the 16S rRNA gene of A. muciniphila and the ttr-region of S. Typhimurium. Calculation of the cell numbers was based on DNA obtained from cell suspensions containing known cell numbers of the targeted bacterial species (see materials and methods). Presence of A. muciniphila in SIHUMI-AS mice is attributed to an increase in the proportion of S. Typhimurium cells at the expense of other community members showing reduced proportion of SIHUMI members. Ten animals per group were used. The bacterial cell numbers and P-values for the differences between the groups are provided in Table 1 .

Techniques Used: Mouse Assay, Infection, Real-time Polymerase Chain Reaction

Presence of both A. muciniphila and S. Typhimurium is accompanied by increased pro-inflammatory cytokines. (A) Cecal mRNA levels of IFN-γ, IP-10, TNF-α, IL-12, IL-6, IL-17 and IL-18 in gnotobiotic SIHUMI mice were measured. mRNA was extracted from cecum mucosa of mice belonging to either one of four groups: SIHUMI, SIHUMI-A, SIHUMI-S and SIHUMI-AS (see Figure. 1). The mRNA was converted to cDNA for quantitative real-time PCR measurement (see materials and methods). Inoculation of the gnotobiotic SIHUMI mice with A. muciniphila followed by S. Typhimurium infection (SIHUMI-AS) caused an increase in mRNA levels of pro-inflammatory cytokines except IL-18. Data are expressed as mean±standard error. n = 6 per group. Star indicates statistically significant differences ( *P
Figure Legend Snippet: Presence of both A. muciniphila and S. Typhimurium is accompanied by increased pro-inflammatory cytokines. (A) Cecal mRNA levels of IFN-γ, IP-10, TNF-α, IL-12, IL-6, IL-17 and IL-18 in gnotobiotic SIHUMI mice were measured. mRNA was extracted from cecum mucosa of mice belonging to either one of four groups: SIHUMI, SIHUMI-A, SIHUMI-S and SIHUMI-AS (see Figure. 1). The mRNA was converted to cDNA for quantitative real-time PCR measurement (see materials and methods). Inoculation of the gnotobiotic SIHUMI mice with A. muciniphila followed by S. Typhimurium infection (SIHUMI-AS) caused an increase in mRNA levels of pro-inflammatory cytokines except IL-18. Data are expressed as mean±standard error. n = 6 per group. Star indicates statistically significant differences ( *P

Techniques Used: Mouse Assay, Real-time Polymerase Chain Reaction, Infection

SIHUMI mice with both A. muciniphila and S. Typhimurium display increased MUC2 mRNA levels (A) and reduced numbers of mucin filled goblet cells (B and C). (A) mRNA was extracted from cecum mucosa of mice belonging to either one of four groups: SIHUMI, SIHUMI-A, SIHUMI-S and SIHUMI-AS. MUC2 mRNA from cecum mucosa was converted to cDNA and expression levels were quantified using real-time PCR (see materials and methods). SIHUMI-A and SIHUMI-AS mice showed significantly higher MUC2 gene expression compared to the other two groups, harboring no A. muciniphila . Data are expressed as mean±standard error. n = 6 per group. *P
Figure Legend Snippet: SIHUMI mice with both A. muciniphila and S. Typhimurium display increased MUC2 mRNA levels (A) and reduced numbers of mucin filled goblet cells (B and C). (A) mRNA was extracted from cecum mucosa of mice belonging to either one of four groups: SIHUMI, SIHUMI-A, SIHUMI-S and SIHUMI-AS. MUC2 mRNA from cecum mucosa was converted to cDNA and expression levels were quantified using real-time PCR (see materials and methods). SIHUMI-A and SIHUMI-AS mice showed significantly higher MUC2 gene expression compared to the other two groups, harboring no A. muciniphila . Data are expressed as mean±standard error. n = 6 per group. *P

Techniques Used: Mouse Assay, Expressing, Real-time Polymerase Chain Reaction

Design of the animal experiment. Fourty C3H mice associated with a defined microbial community of 8 bacterial species (SIHUMI) were allocated to four different groups (10 mice per group). Each mouse was associated with 8 bacterial species (SIHUMI). Twelve weeks-old SIHUMI mice were subsequently associated with A. muciniphila (SIHUMI-A) or S. Typhimurium (SIHUMI-S) or with both A. muciniphila and S. Typhimurium (SIHUMI-AS). SIHUMI mice received only sterile medium. Times of association, infection and killing are as indicated. ‡ - killed.
Figure Legend Snippet: Design of the animal experiment. Fourty C3H mice associated with a defined microbial community of 8 bacterial species (SIHUMI) were allocated to four different groups (10 mice per group). Each mouse was associated with 8 bacterial species (SIHUMI). Twelve weeks-old SIHUMI mice were subsequently associated with A. muciniphila (SIHUMI-A) or S. Typhimurium (SIHUMI-S) or with both A. muciniphila and S. Typhimurium (SIHUMI-AS). SIHUMI mice received only sterile medium. Times of association, infection and killing are as indicated. ‡ - killed.

Techniques Used: Mouse Assay, Infection

2) Product Images from "Beta-Defensin-2 and Beta-Defensin-3 Reduce Intestinal Damage Caused by Salmonella typhimurium Modulating the Expression of Cytokines and Enhancing the Probiotic Activity of Enterococcus faecium"

Article Title: Beta-Defensin-2 and Beta-Defensin-3 Reduce Intestinal Damage Caused by Salmonella typhimurium Modulating the Expression of Cytokines and Enhancing the Probiotic Activity of Enterococcus faecium

Journal: Journal of Immunology Research

doi: 10.1155/2017/6976935

Comparison between relative gene expression (a) and protein concentration (b) in Caco-2 cells infected with S. typhimurium alone and Caco-2 cells coinfected with S. typhimurium and E. faecium . Data are mean ± SD and are expressed as the percentage of increment compared to uninfected controls.
Figure Legend Snippet: Comparison between relative gene expression (a) and protein concentration (b) in Caco-2 cells infected with S. typhimurium alone and Caco-2 cells coinfected with S. typhimurium and E. faecium . Data are mean ± SD and are expressed as the percentage of increment compared to uninfected controls.

Techniques Used: Expressing, Protein Concentration, Infection

S. typhimurium internalization assay. Untransfected Caco-2 cells were infected with S. typhimurium alone or coinfected with live or heat-killed E. faecium for 4 hours. The number of internalized bacteria was determined by host cell lysis, plating, and counting CFU/well. The data shown are representative of three different experiments ± SD. Error bars represent standard deviations.
Figure Legend Snippet: S. typhimurium internalization assay. Untransfected Caco-2 cells were infected with S. typhimurium alone or coinfected with live or heat-killed E. faecium for 4 hours. The number of internalized bacteria was determined by host cell lysis, plating, and counting CFU/well. The data shown are representative of three different experiments ± SD. Error bars represent standard deviations.

Techniques Used: Infection, Lysis

Comparison between relative gene expression (a) and protein concentration (b) in Caco-2 cells infected with S. typhimurium and Caco-2 cells infected with E. faecium . Data are mean ± SD and are expressed as the percentage of increment compared to uninfected controls.
Figure Legend Snippet: Comparison between relative gene expression (a) and protein concentration (b) in Caco-2 cells infected with S. typhimurium and Caco-2 cells infected with E. faecium . Data are mean ± SD and are expressed as the percentage of increment compared to uninfected controls.

Techniques Used: Expressing, Protein Concentration, Infection

3) Product Images from "Enhanced Immunogenicity of Pneumococcal Surface Adhesin A by Genetic Fusion to Cytokines and Evaluation of Protective Immunity in Mice "

Article Title: Enhanced Immunogenicity of Pneumococcal Surface Adhesin A by Genetic Fusion to Cytokines and Evaluation of Protective Immunity in Mice

Journal: Infection and Immunity

doi: 10.1128/IAI.70.10.5589-5595.2002

Flow cytometry was used to demonstrate antigens on the surface of intact S. pneumoniae bacteria. Bacteria grown to log or stationary phase were incubated in 10% serum from naive mice (Ctrl), PsaA-immune mice (anti-PsaA), or type 3 PS-immune mice (anti-PS). Specific binding was visualized using an F(ab′) 2 fragment of goat anti-mouse IgG (H+L) conjugated to Alexa-488 fluorochrome. Bacterium-sized particles were detected using logarithmic side scatter as the threshold, and specific binding was recorded as green fluorescence (FL1). A total of 10 5 events were used to generate each histogram. The histograms are representative of five or three similar experiments performed with log- or stationary-phase bacteria, respectively.
Figure Legend Snippet: Flow cytometry was used to demonstrate antigens on the surface of intact S. pneumoniae bacteria. Bacteria grown to log or stationary phase were incubated in 10% serum from naive mice (Ctrl), PsaA-immune mice (anti-PsaA), or type 3 PS-immune mice (anti-PS). Specific binding was visualized using an F(ab′) 2 fragment of goat anti-mouse IgG (H+L) conjugated to Alexa-488 fluorochrome. Bacterium-sized particles were detected using logarithmic side scatter as the threshold, and specific binding was recorded as green fluorescence (FL1). A total of 10 5 events were used to generate each histogram. The histograms are representative of five or three similar experiments performed with log- or stationary-phase bacteria, respectively.

Techniques Used: Flow Cytometry, Cytometry, Incubation, Mouse Assay, Binding Assay, Fluorescence

4) Product Images from "Functional Transfer of Salmonella Pathogenicity Island 2 to Salmonella bongori and Escherichia coli"

Article Title: Functional Transfer of Salmonella Pathogenicity Island 2 to Salmonella bongori and Escherichia coli

Journal: Infection and Immunity

doi: 10.1128/IAI.72.5.2879-2888.2004

Intracellular proliferation of S. enterica serotype Typhimurium, S. bongori , and S. bongori (pB6). (A) RAW 264.7 macrophages were infected with S. enterica serotype Typhimurium wild type, S. bongori , or S. bongori harboring pB6 encoding the SPI2 locus. All strains harbored p2812 for SPI2-dependent expression of sifA ::M45 and constitutive expression of GFP. Host cells were fixed 16 h after infection, and F-actin was stained by Texas red-phalloidin. Representative micrographs demonstrating the extent of intracellular proliferation are shown. (B) Macrophages were infected at a multiplicity of infection of ∼1 (for S. enterica serotype Typhimurium wild type [S. T. WT]) and 50 (for S. bongori [S. B.] strains), respectively. At 2 and 16 h after infection, macrophages were lysed by the addition of 0.1% Triton X-100, and serial dilutions were plated onto Mueller-Hinton agar for the enumeration of intracellular bacteria. The intracellular CFU was determined for both time points, and the rates of intracellular replication were expressed as the fold increase in CFU. Experiments were performed in triplicate on three different occasions.
Figure Legend Snippet: Intracellular proliferation of S. enterica serotype Typhimurium, S. bongori , and S. bongori (pB6). (A) RAW 264.7 macrophages were infected with S. enterica serotype Typhimurium wild type, S. bongori , or S. bongori harboring pB6 encoding the SPI2 locus. All strains harbored p2812 for SPI2-dependent expression of sifA ::M45 and constitutive expression of GFP. Host cells were fixed 16 h after infection, and F-actin was stained by Texas red-phalloidin. Representative micrographs demonstrating the extent of intracellular proliferation are shown. (B) Macrophages were infected at a multiplicity of infection of ∼1 (for S. enterica serotype Typhimurium wild type [S. T. WT]) and 50 (for S. bongori [S. B.] strains), respectively. At 2 and 16 h after infection, macrophages were lysed by the addition of 0.1% Triton X-100, and serial dilutions were plated onto Mueller-Hinton agar for the enumeration of intracellular bacteria. The intracellular CFU was determined for both time points, and the rates of intracellular replication were expressed as the fold increase in CFU. Experiments were performed in triplicate on three different occasions.

Techniques Used: Infection, Expressing, Staining

Host cell phenotypes induced by S. bongori harboring SPI2. (A) Aggregation of host cell endosomes by S. bongori harboring the SPI2 locus and sifA . HeLa cells were infected with S. enterica serotype Typhimurium, S. bongori , or S. bongori harboring the plasmid-borne SPI2 locus. All strains harbored p2812 for the SPI2-dependent expression of sifA ::M45 and the constitutive expression of GFP. At 16 h after infection, cells were fixed and immunostained for the late endosomal-lysosomal membrane protein LAMP-1. Representative micrographs show the organization of endosomal-lysosomal compartments (red) and intracellular Salmonella spp. Salmonella -induced filaments are indicated by arrowheads. (B) Accumulation of host cell tubulin. HeLa cells were infected with S. bongori , S. bongori harboring the plasmid-borne SPI2 locus, or S. bongori harboring the SPI2 locus and plasmid p2643 for the expression of sseF ::HA. HeLa cells were fixed 16 h after infection and immunostaining was performed for S. bongori lipopolysaccharide (green), β-tubulin (red), and translocated SseF-HA (blue). Alternatively, F-actin was visualized by staining with Texas red-phalloidin (red). Representative cells with accumulation of tubulin at the SCV (indicated by arrows) were shown. Such tubulin accumulations were absent in cells infected with S. bongori or S. bongori harboring only plasmid p2643 (not shown). No accumulation of F-actin at the SCV was observed.
Figure Legend Snippet: Host cell phenotypes induced by S. bongori harboring SPI2. (A) Aggregation of host cell endosomes by S. bongori harboring the SPI2 locus and sifA . HeLa cells were infected with S. enterica serotype Typhimurium, S. bongori , or S. bongori harboring the plasmid-borne SPI2 locus. All strains harbored p2812 for the SPI2-dependent expression of sifA ::M45 and the constitutive expression of GFP. At 16 h after infection, cells were fixed and immunostained for the late endosomal-lysosomal membrane protein LAMP-1. Representative micrographs show the organization of endosomal-lysosomal compartments (red) and intracellular Salmonella spp. Salmonella -induced filaments are indicated by arrowheads. (B) Accumulation of host cell tubulin. HeLa cells were infected with S. bongori , S. bongori harboring the plasmid-borne SPI2 locus, or S. bongori harboring the SPI2 locus and plasmid p2643 for the expression of sseF ::HA. HeLa cells were fixed 16 h after infection and immunostaining was performed for S. bongori lipopolysaccharide (green), β-tubulin (red), and translocated SseF-HA (blue). Alternatively, F-actin was visualized by staining with Texas red-phalloidin (red). Representative cells with accumulation of tubulin at the SCV (indicated by arrows) were shown. Such tubulin accumulations were absent in cells infected with S. bongori or S. bongori harboring only plasmid p2643 (not shown). No accumulation of F-actin at the SCV was observed.

Techniques Used: Infection, Plasmid Preparation, Expressing, Immunostaining, Staining

Expression of sseB by the cloned SPI2 locus. (A) S. enterica serotype Typhimurium wild type (WT), the SPI2 deletion mutant strain (ΔSPI2), and the SPI2 deletion mutant strain complemented with the plasmid-borne SPI2 locus (ΔSPI2 [SPI2]) were grown under conditions resulting in low (LB) or high (PCN-P) levels of the SPI2-encoded protein SseB. After growth for 16 h, lysates were prepared from equal amounts of bacteria, as determined from the OD 600 ). (B) The plasmid-borne SPI2 locus was introduced into S. bongori. S. enterica serotype Typhimurium wild type (S. T.) and S. bongori harboring the vector pBeloBac11 or the SPI2-encoding plasmid pB6 were grown for 16 h in various media as indicated. LB broth was adjusted to pH 7.0 or 5.0, PCN was adjusted to pH 5.8, PCN-P was adjusted to pH 7.4, and F minimal medium containing limiting amounts of Mg 2+ (30 μM) was adjusted to pH 7.0 or 5.0. The amounts of SseB in total bacterial lysates were detected as described above.
Figure Legend Snippet: Expression of sseB by the cloned SPI2 locus. (A) S. enterica serotype Typhimurium wild type (WT), the SPI2 deletion mutant strain (ΔSPI2), and the SPI2 deletion mutant strain complemented with the plasmid-borne SPI2 locus (ΔSPI2 [SPI2]) were grown under conditions resulting in low (LB) or high (PCN-P) levels of the SPI2-encoded protein SseB. After growth for 16 h, lysates were prepared from equal amounts of bacteria, as determined from the OD 600 ). (B) The plasmid-borne SPI2 locus was introduced into S. bongori. S. enterica serotype Typhimurium wild type (S. T.) and S. bongori harboring the vector pBeloBac11 or the SPI2-encoding plasmid pB6 were grown for 16 h in various media as indicated. LB broth was adjusted to pH 7.0 or 5.0, PCN was adjusted to pH 5.8, PCN-P was adjusted to pH 7.4, and F minimal medium containing limiting amounts of Mg 2+ (30 μM) was adjusted to pH 7.0 or 5.0. The amounts of SseB in total bacterial lysates were detected as described above.

Techniques Used: Expressing, Clone Assay, Mutagenesis, Plasmid Preparation

Translocation of a SPI2 effector protein by intracellular S. enterica serotype Typhimurium and by S. bongori and E. coli harboring the SPI2-encoded type III secretion system. RAW 264.7 cells were grown on glass coverslips and infected with S. enterica serotype Typhimurium, S. bongori , or E. coli Mutaflor. All strains harbored plasmid p2777 for SPI2-regulated expression of sseJ ::HA and constitutive expression of GFP. In addition, S. bongori or E. coli Mutaflor harbored pB6 containing the SPI2 locus, if indicated. At 16 h after infection, the cells were fixed and immunostained for SseJ-HA. Representative micrographs show merged phase contrast, intracellular bacteria (green), and translocated SseJ-HA (red).
Figure Legend Snippet: Translocation of a SPI2 effector protein by intracellular S. enterica serotype Typhimurium and by S. bongori and E. coli harboring the SPI2-encoded type III secretion system. RAW 264.7 cells were grown on glass coverslips and infected with S. enterica serotype Typhimurium, S. bongori , or E. coli Mutaflor. All strains harbored plasmid p2777 for SPI2-regulated expression of sseJ ::HA and constitutive expression of GFP. In addition, S. bongori or E. coli Mutaflor harbored pB6 containing the SPI2 locus, if indicated. At 16 h after infection, the cells were fixed and immunostained for SseJ-HA. Representative micrographs show merged phase contrast, intracellular bacteria (green), and translocated SseJ-HA (red).

Techniques Used: Translocation Assay, Infection, Plasmid Preparation, Expressing

5) Product Images from "Killing of Candida albicans Filaments by Salmonella enterica Serovar Typhimurium Is Mediated by sopB Effectors, Parts of a Type III Secretion System ▿"

Article Title: Killing of Candida albicans Filaments by Salmonella enterica Serovar Typhimurium Is Mediated by sopB Effectors, Parts of a Type III Secretion System ▿

Journal: Eukaryotic Cell

doi: 10.1128/EC.00014-11

Gene expression in S. Typhimurium and C. albicans . (A) Transcription of sopB and sipB of the S. Typhimurium wild type in the presence of supernatants from filaments or yeast-form C. albicans . (B) Transcriptional levels of TEC1 , HWP1 , ALS3 , and CDC42 in
Figure Legend Snippet: Gene expression in S. Typhimurium and C. albicans . (A) Transcription of sopB and sipB of the S. Typhimurium wild type in the presence of supernatants from filaments or yeast-form C. albicans . (B) Transcriptional levels of TEC1 , HWP1 , ALS3 , and CDC42 in

Techniques Used: Expressing

S. Typhimurium kills C. albicans filaments via the sopB effector. Viability of C. albicans filaments using the XTT assay. (A) C. albicans Δ tup1 or Δ suv3 strains were cultured at 25°C for 48 h for constitutive filament production
Figure Legend Snippet: S. Typhimurium kills C. albicans filaments via the sopB effector. Viability of C. albicans filaments using the XTT assay. (A) C. albicans Δ tup1 or Δ suv3 strains were cultured at 25°C for 48 h for constitutive filament production

Techniques Used: XTT Assay, Cell Culture

The S. Typhimurium sopB effector influences the viability of C. albicans under an in vitro planktonic environment. The viability of C. albicans in the presence of S. Typhimurium wild type (WT) or various mutants was determined at 25°C after 5
Figure Legend Snippet: The S. Typhimurium sopB effector influences the viability of C. albicans under an in vitro planktonic environment. The viability of C. albicans in the presence of S. Typhimurium wild type (WT) or various mutants was determined at 25°C after 5

Techniques Used: In Vitro

S. Typhimurium strongly inhibits C. albicans filamentation and biofilm formation via the sopB effector. (A) Attachment of S. Typhimurium wild-type (WT) and sopB mutant strains to the C. albicans filaments. C. albicans SSY50-B filaments were formed in
Figure Legend Snippet: S. Typhimurium strongly inhibits C. albicans filamentation and biofilm formation via the sopB effector. (A) Attachment of S. Typhimurium wild-type (WT) and sopB mutant strains to the C. albicans filaments. C. albicans SSY50-B filaments were formed in

Techniques Used: Mutagenesis

Translocation of SopB into C. albicans filaments via SipB. SopB translocation in S. Typhimurium wild-type (top row) and sipB mutant (bottom row) was visualized by confocal microscopy. Scale bars, 4.2 μm. The inset shows an enlarged view of the
Figure Legend Snippet: Translocation of SopB into C. albicans filaments via SipB. SopB translocation in S. Typhimurium wild-type (top row) and sipB mutant (bottom row) was visualized by confocal microscopy. Scale bars, 4.2 μm. The inset shows an enlarged view of the

Techniques Used: Translocation Assay, Mutagenesis, Confocal Microscopy

6) Product Images from "Porins from Salmonella enterica Serovar Typhimurium Activate the Transcription Factors Activating Protein 1 and NF-?B through the Raf-1-Mitogen-Activated Protein Kinase Cascade "

Article Title: Porins from Salmonella enterica Serovar Typhimurium Activate the Transcription Factors Activating Protein 1 and NF-?B through the Raf-1-Mitogen-Activated Protein Kinase Cascade

Journal: Infection and Immunity

doi: 10.1128/IAI.70.2.558-568.2002

Effect of S. enterica serovar Typhimurium ( S.ty ) porins on Raf-1 (A) and MEK1/2 (B) activation after different stimulation times. Proteins from cell lysates (3 × 10 6 cells/ml) were analyzed by SDS-15% PAGE and immunoblotted with a Raf-1- or MEK1/2-phosphospecific antibody. Shifts in band mobility on SDS-PAGE due to phosphorylation were obtained with anti-Raf-1, anti-MEK1, and anti-MEK2. Fold increases in Raf-1 and MEK1/2 activation, compared to unstimulated cells (control), are shown below each lane for each blot. BSA represents a control of an unspecific stimulus.
Figure Legend Snippet: Effect of S. enterica serovar Typhimurium ( S.ty ) porins on Raf-1 (A) and MEK1/2 (B) activation after different stimulation times. Proteins from cell lysates (3 × 10 6 cells/ml) were analyzed by SDS-15% PAGE and immunoblotted with a Raf-1- or MEK1/2-phosphospecific antibody. Shifts in band mobility on SDS-PAGE due to phosphorylation were obtained with anti-Raf-1, anti-MEK1, and anti-MEK2. Fold increases in Raf-1 and MEK1/2 activation, compared to unstimulated cells (control), are shown below each lane for each blot. BSA represents a control of an unspecific stimulus.

Techniques Used: Activation Assay, Polyacrylamide Gel Electrophoresis, SDS Page

Pattern of S. enterica serovar Typhimurium SH5014 porins and LPS on SDS-PAGE. (A) Lane 1, molecular mass standards (Amersham Pharmacia Biotech, Milan, Italy) (phosphorylase b, 94 kDa,; albumin, 67 kDa; ovalbumin, 43 kDa; carbonic anhydrase, 30 kDa; trypsin inhibitor, 20.1 kDa; α-lactalbumin, 14.4 kDa); lane 2, S. enterica serovar Typhimurium porins (10 μg). (B) Lane 1, S. enterica serovar Typhimurium LPS standard (Sigma-Aldrich S.r.l.); lane 2, S. enterica serovar Typhimurium SH5014 LPS (10 μg).
Figure Legend Snippet: Pattern of S. enterica serovar Typhimurium SH5014 porins and LPS on SDS-PAGE. (A) Lane 1, molecular mass standards (Amersham Pharmacia Biotech, Milan, Italy) (phosphorylase b, 94 kDa,; albumin, 67 kDa; ovalbumin, 43 kDa; carbonic anhydrase, 30 kDa; trypsin inhibitor, 20.1 kDa; α-lactalbumin, 14.4 kDa); lane 2, S. enterica serovar Typhimurium porins (10 μg). (B) Lane 1, S. enterica serovar Typhimurium LPS standard (Sigma-Aldrich S.r.l.); lane 2, S. enterica serovar Typhimurium SH5014 LPS (10 μg).

Techniques Used: SDS Page

Effect of S. enterica serovar Typhimurium ( S.ty ) porins on ERK1/2 (A), p38 (B), and JNK (C) activation after different stimulation times. Proteins from cell lysates (3 × 10 6 cells/ml) were analyzed by SDS-15% PAGE and immunoblotted with an ERK1/2-, p38-, or JNK-phosphospecific antibody. Shifts in band mobility on SDS-PAGE due to phosphorylation were obtained with anti-ERK1/2, anti-p38, and anti-JNK. ERK1 appears as the upper band (44 kDa); ERK2 appears as the lower band (42 kDa). Fold increases in ERK1/2, p38, and JNK activation are shown below each lane for each blot. BSA represents a control of an unspecific stimulus.
Figure Legend Snippet: Effect of S. enterica serovar Typhimurium ( S.ty ) porins on ERK1/2 (A), p38 (B), and JNK (C) activation after different stimulation times. Proteins from cell lysates (3 × 10 6 cells/ml) were analyzed by SDS-15% PAGE and immunoblotted with an ERK1/2-, p38-, or JNK-phosphospecific antibody. Shifts in band mobility on SDS-PAGE due to phosphorylation were obtained with anti-ERK1/2, anti-p38, and anti-JNK. ERK1 appears as the upper band (44 kDa); ERK2 appears as the lower band (42 kDa). Fold increases in ERK1/2, p38, and JNK activation are shown below each lane for each blot. BSA represents a control of an unspecific stimulus.

Techniques Used: Activation Assay, Polyacrylamide Gel Electrophoresis, SDS Page

Western blotting analysis with pJNK and phospho-p38 antibodies of lysates from C3H/HeJ peritoneal macrophages (5 × 10 6 cells/ml) after stimulation with LPS (1 μg/ml) or porins (5 μg/ml) for 20 min. The experimental procedure was as described in Materials and Methods. S. ty ., S. enterica serovar Typhimurium.
Figure Legend Snippet: Western blotting analysis with pJNK and phospho-p38 antibodies of lysates from C3H/HeJ peritoneal macrophages (5 × 10 6 cells/ml) after stimulation with LPS (1 μg/ml) or porins (5 μg/ml) for 20 min. The experimental procedure was as described in Materials and Methods. S. ty ., S. enterica serovar Typhimurium.

Techniques Used: Western Blot

7) Product Images from "Genetics of Swarming Motility in Salmonella enterica Serovar Typhimurium: Critical Role for Lipopolysaccharide"

Article Title: Genetics of Swarming Motility in Salmonella enterica Serovar Typhimurium: Critical Role for Lipopolysaccharide

Journal: Journal of Bacteriology

doi:

Rescue of swarming defects with external addition of surfactin and LPS. Five-microliter drops of solution containing surfactin or LPS were placed at the center of 0.5% Difco swarm medium, inoculated with S. enterica serovar Typhimurium ( waaL and cheA ) and E. coli (RP437) strains, and incubated overnight as described in Materials and Methods.
Figure Legend Snippet: Rescue of swarming defects with external addition of surfactin and LPS. Five-microliter drops of solution containing surfactin or LPS were placed at the center of 0.5% Difco swarm medium, inoculated with S. enterica serovar Typhimurium ( waaL and cheA ) and E. coli (RP437) strains, and incubated overnight as described in Materials and Methods.

Techniques Used: Incubation

8) Product Images from "Genetics of Swarming Motility in Salmonella enterica Serovar Typhimurium: Critical Role for Lipopolysaccharide"

Article Title: Genetics of Swarming Motility in Salmonella enterica Serovar Typhimurium: Critical Role for Lipopolysaccharide

Journal: Journal of Bacteriology

doi:

Rescue of swarming defects with external addition of surfactin and LPS. Five-microliter drops of solution containing surfactin or LPS were placed at the center of 0.5% Difco swarm medium, inoculated with S. enterica serovar Typhimurium ( waaL and cheA ) and E. coli (RP437) strains, and incubated overnight as described in Materials and Methods.
Figure Legend Snippet: Rescue of swarming defects with external addition of surfactin and LPS. Five-microliter drops of solution containing surfactin or LPS were placed at the center of 0.5% Difco swarm medium, inoculated with S. enterica serovar Typhimurium ( waaL and cheA ) and E. coli (RP437) strains, and incubated overnight as described in Materials and Methods.

Techniques Used: Incubation

Precocious swarming of waaG LPS core mutant. (A) A wild-type (wt) S. enterica serovar Typhimurium strain (SL3770) and its waaG mutant derivative (SL3769) were inoculated on Difco Bacto swarm medium and incubated at 37°C. Photographs were taken after 5 and 20 h of incubation. The arrow points to edge of the waaG swarm colony. (B) The waaG strain was inoculated on minimal swarm medium and incubated at 37°C. Photographs were taken after 24, 30, and 44 h. The arrow indicates edge of the slime ring. Swarming movement was confined to regions that appear as branches emanating from the center.
Figure Legend Snippet: Precocious swarming of waaG LPS core mutant. (A) A wild-type (wt) S. enterica serovar Typhimurium strain (SL3770) and its waaG mutant derivative (SL3769) were inoculated on Difco Bacto swarm medium and incubated at 37°C. Photographs were taken after 5 and 20 h of incubation. The arrow points to edge of the waaG swarm colony. (B) The waaG strain was inoculated on minimal swarm medium and incubated at 37°C. Photographs were taken after 24, 30, and 44 h. The arrow indicates edge of the slime ring. Swarming movement was confined to regions that appear as branches emanating from the center.

Techniques Used: Mutagenesis, Incubation

9) Product Images from "Contribution of Salmonella typhimurium Virulence Factors to Diarrheal Disease in Calves"

Article Title: Contribution of Salmonella typhimurium Virulence Factors to Diarrheal Disease in Calves

Journal: Infection and Immunity

doi:

Histopathology of Peyer’s patches and ileum from perinatal calves inoculated per os with 10 10 CFU of the indicated strains of S. typhimurium per animal, photographed at a 40× magnification. Shown are hematoxylin-and-eosin-stained sections of Peyer’s patch (left column) and terminal ileum (right column). Short arrows indicate areas of marked lymphoid depletion; long arrows indicate zones of variable degrees of fibrinopurulent necrotizing ileitis at the mucosal surface.
Figure Legend Snippet: Histopathology of Peyer’s patches and ileum from perinatal calves inoculated per os with 10 10 CFU of the indicated strains of S. typhimurium per animal, photographed at a 40× magnification. Shown are hematoxylin-and-eosin-stained sections of Peyer’s patch (left column) and terminal ileum (right column). Short arrows indicate areas of marked lymphoid depletion; long arrows indicate zones of variable degrees of fibrinopurulent necrotizing ileitis at the mucosal surface.

Techniques Used: Histopathology, Staining

Representative examples of the gross pathology of Peyer’s patch and mucosa of the terminal ileum of calves inoculated orally with 10 10 CFU of different S. typhimurium strains. (A) Severe acute fibrinopurulent necrotizing enteritis with segmental or continuous pseudomembrane formation of calves inoculated with wild type (IR715), strain STN272 ( spvR ), or strain SVM255 ( sigD ). (B) Moderate to marked subacute fibrinopurulent enteritis often confined to Peyer’s patches of calves inoculated with STN119 ( spiB ) or STN166 ( rfaJ ). (C) Normal Peyer’s patch and ileal mucosa of calves inoculated with STN61 ( hilA ), STN162 ( prgH ), or CL1509 ( aroA ) or of uninoculated controls. Bar = 1 cm.
Figure Legend Snippet: Representative examples of the gross pathology of Peyer’s patch and mucosa of the terminal ileum of calves inoculated orally with 10 10 CFU of different S. typhimurium strains. (A) Severe acute fibrinopurulent necrotizing enteritis with segmental or continuous pseudomembrane formation of calves inoculated with wild type (IR715), strain STN272 ( spvR ), or strain SVM255 ( sigD ). (B) Moderate to marked subacute fibrinopurulent enteritis often confined to Peyer’s patches of calves inoculated with STN119 ( spiB ) or STN166 ( rfaJ ). (C) Normal Peyer’s patch and ileal mucosa of calves inoculated with STN61 ( hilA ), STN162 ( prgH ), or CL1509 ( aroA ) or of uninoculated controls. Bar = 1 cm.

Techniques Used:

10) Product Images from "Osteoprotegerin Regulates Pancreatic β-Cell Homeostasis upon Microbial Invasion"

Article Title: Osteoprotegerin Regulates Pancreatic β-Cell Homeostasis upon Microbial Invasion

Journal: PLoS ONE

doi: 10.1371/journal.pone.0146544

Bone homeostasis in mice after Salmonella infection. (A) TRAP staining of tibial sections of BALB/c mice infected for 5 days with Salmonella enterica . Cont, uninfected mice; PsB, periosteal bone; TB, trabecular bone. Scale bars in upper panels represent 500 μm, and in middle or lower panels, 200 μm. (B, C) The number of osteoclasts (N.OC) at the trabecular bone surface (TBS) and the periosteal bone surface (PsBS) in tibial sections. Shown are means ± SD. Sal, Salmonella -infected mice. (D, E) Serum OPG levels (D) or RANKL levels (E) in mice infected one week with the avirulent Salmonella enterica strains UF20, UF71 and UF110 (n = 6 for each group). Open circles indicate outliers. *** P
Figure Legend Snippet: Bone homeostasis in mice after Salmonella infection. (A) TRAP staining of tibial sections of BALB/c mice infected for 5 days with Salmonella enterica . Cont, uninfected mice; PsB, periosteal bone; TB, trabecular bone. Scale bars in upper panels represent 500 μm, and in middle or lower panels, 200 μm. (B, C) The number of osteoclasts (N.OC) at the trabecular bone surface (TBS) and the periosteal bone surface (PsBS) in tibial sections. Shown are means ± SD. Sal, Salmonella -infected mice. (D, E) Serum OPG levels (D) or RANKL levels (E) in mice infected one week with the avirulent Salmonella enterica strains UF20, UF71 and UF110 (n = 6 for each group). Open circles indicate outliers. *** P

Techniques Used: Mouse Assay, Infection, Staining

11) Product Images from "Induction and Resuscitation of Viable but Nonculturable Salmonella enterica Serovar Typhimurium DT104+ "

Article Title: Induction and Resuscitation of Viable but Nonculturable Salmonella enterica Serovar Typhimurium DT104+

Journal: Applied and Environmental Microbiology

doi: 10.1128/AEM.69.11.6669-6675.2003

Comparison of nonculturable S. enterica serovar Typhimurium DT104 11601 (∼5 × 10 4 CFU/ml) exposed to an 80°C (56°C internal temperature) upshift for 15 s (left) versus no temperature upshift (right) followed by inoculation on TSA at 37°C for 24 h. Selected colonies from the left plate were tested with Gram stain, TSI agar slants, and API 20E and were confirmed to be Salmonella .
Figure Legend Snippet: Comparison of nonculturable S. enterica serovar Typhimurium DT104 11601 (∼5 × 10 4 CFU/ml) exposed to an 80°C (56°C internal temperature) upshift for 15 s (left) versus no temperature upshift (right) followed by inoculation on TSA at 37°C for 24 h. Selected colonies from the left plate were tested with Gram stain, TSI agar slants, and API 20E and were confirmed to be Salmonella .

Techniques Used: Staining

Comparison of the survival of freshly cultured (gray bars) (∼3 × 10 4 CFU/ml) and nutrient-deficient S. enterica serovar Typhimurium DT104 11601 (∼3 × 10 4 CFU/ml) (black bars) maintained in 7.35 mM BPS microcosms at 21°C for 418 days, after exposure to10 mM H 2 O 2 solution for 100 min. Counts represent the mean of replicate experiments. Points shown on the baseline represent undetectable populations after standard plate count determinations.
Figure Legend Snippet: Comparison of the survival of freshly cultured (gray bars) (∼3 × 10 4 CFU/ml) and nutrient-deficient S. enterica serovar Typhimurium DT104 11601 (∼3 × 10 4 CFU/ml) (black bars) maintained in 7.35 mM BPS microcosms at 21°C for 418 days, after exposure to10 mM H 2 O 2 solution for 100 min. Counts represent the mean of replicate experiments. Points shown on the baseline represent undetectable populations after standard plate count determinations.

Techniques Used: Cell Culture

Scanning electron micrographs of S. enterica serovar Typhimurium DT104 11601 exposed to nutrient-limiting conditions for more than 365 days (A) versus cells grown for 24 h in TSB (B). Scale bar, 1 μm.
Figure Legend Snippet: Scanning electron micrographs of S. enterica serovar Typhimurium DT104 11601 exposed to nutrient-limiting conditions for more than 365 days (A) versus cells grown for 24 h in TSB (B). Scale bar, 1 μm.

Techniques Used:

Comparison of the recovery of nutrient-deprived S. enterica serovar Typhimurium DT104 11601 previously maintained in microcosms containing 2.3% Instant Ocean at 5°C (○) and 21°C (▴) on catalase-supplemented (broken line) versus nonsupplemented (solid line) TSA plates. Comparable results were obtained with similar microcosms that had been maintained at 37°C for the same period of time. Points shown on the baseline represent undetectable populations after standard plate count determinations.
Figure Legend Snippet: Comparison of the recovery of nutrient-deprived S. enterica serovar Typhimurium DT104 11601 previously maintained in microcosms containing 2.3% Instant Ocean at 5°C (○) and 21°C (▴) on catalase-supplemented (broken line) versus nonsupplemented (solid line) TSA plates. Comparable results were obtained with similar microcosms that had been maintained at 37°C for the same period of time. Points shown on the baseline represent undetectable populations after standard plate count determinations.

Techniques Used:

Effect of nutrient limitation on S. enterica serovar Typhimurium DT104 11601 maintained in microcosms at 21°C (full line) containing 7.35 mM (•) and 0.0735 (▪) mM BPS and at 5°C (broken line) in 7.35 (□) and 0.0735 (○) mM BPS over 425 days. Each data point represents a single measurement of the number of CFU per milliliter. Comparable results were obtained with microscosms containing 0.735 mM BPS when the cells were maintained at 5 and 21°C and held for the same period of time (data not shown). Points shown on the baseline represent undetectable populations after standard plate count determinations.
Figure Legend Snippet: Effect of nutrient limitation on S. enterica serovar Typhimurium DT104 11601 maintained in microcosms at 21°C (full line) containing 7.35 mM (•) and 0.0735 (▪) mM BPS and at 5°C (broken line) in 7.35 (□) and 0.0735 (○) mM BPS over 425 days. Each data point represents a single measurement of the number of CFU per milliliter. Comparable results were obtained with microscosms containing 0.735 mM BPS when the cells were maintained at 5 and 21°C and held for the same period of time (data not shown). Points shown on the baseline represent undetectable populations after standard plate count determinations.

Techniques Used:

Scanning electron micrograph of representative S. enterica serovar Typhimurium DT104 11601 exhibiting bacillary and coccoid cell morphologies after exposure to nutrient-limiting conditions for more than 365 days. Scale bar, 1 μm.
Figure Legend Snippet: Scanning electron micrograph of representative S. enterica serovar Typhimurium DT104 11601 exhibiting bacillary and coccoid cell morphologies after exposure to nutrient-limiting conditions for more than 365 days. Scale bar, 1 μm.

Techniques Used:

Comparison of the survival of a freshly cultured suspension (∼10 4 CFU/ml) (▴) and a nutrient-deficient suspension of S. enterica serovar Typhimurium DT104 11601 maintained in 7.35 mM BPS microcosms at 21°C (▪) and 5°C (○) for 178 days after exposure to saturated NaCl for 22 to 168 h. Each data point represents a single measurement of the number of CFU per milliliter.
Figure Legend Snippet: Comparison of the survival of a freshly cultured suspension (∼10 4 CFU/ml) (▴) and a nutrient-deficient suspension of S. enterica serovar Typhimurium DT104 11601 maintained in 7.35 mM BPS microcosms at 21°C (▪) and 5°C (○) for 178 days after exposure to saturated NaCl for 22 to 168 h. Each data point represents a single measurement of the number of CFU per milliliter.

Techniques Used: Cell Culture

Resuscitation of nonculturable S. enterica serovar Typhimurium DT104 11601 maintained in 7.35 mM BPS at 5°C for 273 days when 5.0 ml of a concentration of ∼10 5 CFU/ml was added to 5.0 ml of brain heart infusion broth (final concentration, 5 × 10 4 CFU/ml) and subjected to temperature upshifts to 21°C ( X ), 25°C (▪), and 37°C (•). Cultures maintained at 5°C (♦) without temperature upshift served as controls. Counts represent the mean of replicates. Except for the cultures subjected to a 37°C upshift, the timelines for the other cultures remained on the baseline. The points shown on the baseline represent undetectable populations after standard plate count determinations.
Figure Legend Snippet: Resuscitation of nonculturable S. enterica serovar Typhimurium DT104 11601 maintained in 7.35 mM BPS at 5°C for 273 days when 5.0 ml of a concentration of ∼10 5 CFU/ml was added to 5.0 ml of brain heart infusion broth (final concentration, 5 × 10 4 CFU/ml) and subjected to temperature upshifts to 21°C ( X ), 25°C (▪), and 37°C (•). Cultures maintained at 5°C (♦) without temperature upshift served as controls. Counts represent the mean of replicates. Except for the cultures subjected to a 37°C upshift, the timelines for the other cultures remained on the baseline. The points shown on the baseline represent undetectable populations after standard plate count determinations.

Techniques Used: Concentration Assay

12) Product Images from "Increased Resistance of Salmonella enterica Serovar Typhimurium and Escherichia coli O157:H7 to 222-Nanometer Krypton-Chlorine Excilamp Treatment by Acid Adaptation"

Article Title: Increased Resistance of Salmonella enterica Serovar Typhimurium and Escherichia coli O157:H7 to 222-Nanometer Krypton-Chlorine Excilamp Treatment by Acid Adaptation

Journal: Applied and Environmental Microbiology

doi: 10.1128/AEM.02221-18

Destruction of cell membrane of non-acid-adapted or acid-adapted Salmonella Typhimurium (A) or Escherichia coli O157:H7 (B) as determined by the PI uptake assay. Data represent averages for three independent experiments, and error bars indicate standard deviations. The data were normalized by subtracting fluorescence values obtained for untreated cells and against the OD 600 , as follows: (fluorescence value after treatment − fluorescence value for untreated control)/OD 600 . Different uppercase letters for cells grown in the same matrix indicate significant differences ( P
Figure Legend Snippet: Destruction of cell membrane of non-acid-adapted or acid-adapted Salmonella Typhimurium (A) or Escherichia coli O157:H7 (B) as determined by the PI uptake assay. Data represent averages for three independent experiments, and error bars indicate standard deviations. The data were normalized by subtracting fluorescence values obtained for untreated cells and against the OD 600 , as follows: (fluorescence value after treatment − fluorescence value for untreated control)/OD 600 . Different uppercase letters for cells grown in the same matrix indicate significant differences ( P

Techniques Used: Fluorescence

Lipid peroxidation of cell membrane of non-acid-adapted or acid-adapted Salmonella Typhimurium (A) or Escherichia coli O157:H7 (B) as determined by the DPPP assay. Data represent averages for three independent experiments, and error bars indicate standard deviations. The data were normalized by subtracting fluorescence values obtained for untreated cells and against the OD 600 , as follows: (fluorescence value after treatment − fluorescence value for untreated control)/OD 600 . Different uppercase letters for cells grown in the same matrix indicate significant differences ( P
Figure Legend Snippet: Lipid peroxidation of cell membrane of non-acid-adapted or acid-adapted Salmonella Typhimurium (A) or Escherichia coli O157:H7 (B) as determined by the DPPP assay. Data represent averages for three independent experiments, and error bars indicate standard deviations. The data were normalized by subtracting fluorescence values obtained for untreated cells and against the OD 600 , as follows: (fluorescence value after treatment − fluorescence value for untreated control)/OD 600 . Different uppercase letters for cells grown in the same matrix indicate significant differences ( P

Techniques Used: Fluorescence

13) Product Images from "Genetics of Swarming Motility in Salmonella enterica Serovar Typhimurium: Critical Role for Lipopolysaccharide"

Article Title: Genetics of Swarming Motility in Salmonella enterica Serovar Typhimurium: Critical Role for Lipopolysaccharide

Journal: Journal of Bacteriology

doi:

Rescue of swarming defects with external addition of surfactin and LPS. Five-microliter drops of solution containing surfactin or LPS were placed at the center of 0.5% Difco swarm medium, inoculated with S. enterica serovar Typhimurium ( waaL and cheA ) and E. coli (RP437) strains, and incubated overnight as described in Materials and Methods.
Figure Legend Snippet: Rescue of swarming defects with external addition of surfactin and LPS. Five-microliter drops of solution containing surfactin or LPS were placed at the center of 0.5% Difco swarm medium, inoculated with S. enterica serovar Typhimurium ( waaL and cheA ) and E. coli (RP437) strains, and incubated overnight as described in Materials and Methods.

Techniques Used: Incubation

Precocious swarming of waaG LPS core mutant. (A) A wild-type (wt) S. enterica serovar Typhimurium strain (SL3770) and its waaG mutant derivative (SL3769) were inoculated on Difco Bacto swarm medium and incubated at 37°C. Photographs were taken after 5 and 20 h of incubation. The arrow points to edge of the waaG swarm colony. (B) The waaG strain was inoculated on minimal swarm medium and incubated at 37°C. Photographs were taken after 24, 30, and 44 h. The arrow indicates edge of the slime ring. Swarming movement was confined to regions that appear as branches emanating from the center.
Figure Legend Snippet: Precocious swarming of waaG LPS core mutant. (A) A wild-type (wt) S. enterica serovar Typhimurium strain (SL3770) and its waaG mutant derivative (SL3769) were inoculated on Difco Bacto swarm medium and incubated at 37°C. Photographs were taken after 5 and 20 h of incubation. The arrow points to edge of the waaG swarm colony. (B) The waaG strain was inoculated on minimal swarm medium and incubated at 37°C. Photographs were taken after 24, 30, and 44 h. The arrow indicates edge of the slime ring. Swarming movement was confined to regions that appear as branches emanating from the center.

Techniques Used: Mutagenesis, Incubation

14) Product Images from "Glycerol Supplementation Enhances L. reuteri's Protective Effect against S. Typhimurium Colonization in a 3-D Model of Colonic Epithelium"

Article Title: Glycerol Supplementation Enhances L. reuteri's Protective Effect against S. Typhimurium Colonization in a 3-D Model of Colonic Epithelium

Journal: PLoS ONE

doi: 10.1371/journal.pone.0037116

The L. reuteri supernatants affected the S. Typhimurium χ3339 population stronger in the presence of the 3-D HT-29 aggregates. Log counts (CFU.mL −1 ) of the χ3339 population (initial inoculum 2×10 6 cells.mL −1 ) exposed to the 10-fold diluted supernatant without (SN- 1∶10) and with reuterin (SN+1∶10, 2.5 mM 3-HPA) both in the absence (left) and presence (right) of 3-D HT-29 aggregates. Detection limit = 10 2 CFU.mL −1 .
Figure Legend Snippet: The L. reuteri supernatants affected the S. Typhimurium χ3339 population stronger in the presence of the 3-D HT-29 aggregates. Log counts (CFU.mL −1 ) of the χ3339 population (initial inoculum 2×10 6 cells.mL −1 ) exposed to the 10-fold diluted supernatant without (SN- 1∶10) and with reuterin (SN+1∶10, 2.5 mM 3-HPA) both in the absence (left) and presence (right) of 3-D HT-29 aggregates. Detection limit = 10 2 CFU.mL −1 .

Techniques Used:

The presence of an established L. reuteri population reduced χ3339 adhesion and invasion in 3-D HT-29 aggregates, and this effect was significantly higher when L. reuteri was producing reuterin in situ . Log differences in χ3339 counts (CFU.mL −1 ) of (A) adhered and invaded minus initial inoculum, (B) intracellular surviving minus adhered and invaded, and (C) intracellular growth minus survival upon exposure to an established population of L. reuteri ATCC PTA 6475 producing reuterin (PTA 6475+40 mM gly) or unable to produce reuterin (Δ pduC +40 mM gly and PTA 6475 no gly). Significant differences between the treatments are indicated with different letters (a, b or c; p
Figure Legend Snippet: The presence of an established L. reuteri population reduced χ3339 adhesion and invasion in 3-D HT-29 aggregates, and this effect was significantly higher when L. reuteri was producing reuterin in situ . Log differences in χ3339 counts (CFU.mL −1 ) of (A) adhered and invaded minus initial inoculum, (B) intracellular surviving minus adhered and invaded, and (C) intracellular growth minus survival upon exposure to an established population of L. reuteri ATCC PTA 6475 producing reuterin (PTA 6475+40 mM gly) or unable to produce reuterin (Δ pduC +40 mM gly and PTA 6475 no gly). Significant differences between the treatments are indicated with different letters (a, b or c; p

Techniques Used: In Situ

Experimental procedure for infection of 3-D HT-29 aggregates with S. Typhimurium χ3339 (red line). Approach 1 infection experiments were done in the presence of supernatants of the L. reuteri ferment with or without reuterin (green star), while approach 2 infections were done in the presence of an established L. reuteri population (blue line) producing reuterin in situ .
Figure Legend Snippet: Experimental procedure for infection of 3-D HT-29 aggregates with S. Typhimurium χ3339 (red line). Approach 1 infection experiments were done in the presence of supernatants of the L. reuteri ferment with or without reuterin (green star), while approach 2 infections were done in the presence of an established L. reuteri population (blue line) producing reuterin in situ .

Techniques Used: Infection, In Situ

Long-term (24 h) exposure of 3-D HT-29 aggregates to 3-HPA (2.5–3 mM) results in loss of cell-cell contact and cell viability. (A) Aggregates of HT-29 colon cancer cells grown in three dimensions on porous collagen-coated microcarrier beads. The picture represents the situation for all treatments after 0, 1 and 4 h and the treatments with the L. reuteri supernatants without reuterin (SN- 1∶10) or containing low concentrations of reuterin (SN+1∶100 with 0.25 mM 3-HPA and SN+1∶1000 with 0.025 mM 3-HPA) after 24 h. (B) Example of a single cell suspension and naked porous collagen-coated microcarrier beads after 24 h of exposure to 3 mM 3-HPA or the 10-fold diluted reuterin-containing L. reuteri supernatant (SN+1∶10, 2.5 mM 3-HPA). (C) Example of live/dead counts of trypan blue treated single cell suspensions depicted in B with a hemocytometer. Dead cells are colored blue.
Figure Legend Snippet: Long-term (24 h) exposure of 3-D HT-29 aggregates to 3-HPA (2.5–3 mM) results in loss of cell-cell contact and cell viability. (A) Aggregates of HT-29 colon cancer cells grown in three dimensions on porous collagen-coated microcarrier beads. The picture represents the situation for all treatments after 0, 1 and 4 h and the treatments with the L. reuteri supernatants without reuterin (SN- 1∶10) or containing low concentrations of reuterin (SN+1∶100 with 0.25 mM 3-HPA and SN+1∶1000 with 0.025 mM 3-HPA) after 24 h. (B) Example of a single cell suspension and naked porous collagen-coated microcarrier beads after 24 h of exposure to 3 mM 3-HPA or the 10-fold diluted reuterin-containing L. reuteri supernatant (SN+1∶10, 2.5 mM 3-HPA). (C) Example of live/dead counts of trypan blue treated single cell suspensions depicted in B with a hemocytometer. Dead cells are colored blue.

Techniques Used:

15) Product Images from "Functional Analysis of Lactobacillus rhamnosus GG Pili in Relation to Adhesion and Immunomodulatory Interactions with Intestinal Epithelial Cells"

Article Title: Functional Analysis of Lactobacillus rhamnosus GG Pili in Relation to Adhesion and Immunomodulatory Interactions with Intestinal Epithelial Cells

Journal: Applied and Environmental Microbiology

doi: 10.1128/AEM.06192-11

Comparison of biofilm formation of wild-type L. rhamnosus GG (LGG WT) and its knockout mutant derivatives. Formation of L. rhamnosus GG biofilms on polystyrene pegs occurring after 72 h of incubation in lactobacilli AOAC medium was quantified by crystal
Figure Legend Snippet: Comparison of biofilm formation of wild-type L. rhamnosus GG (LGG WT) and its knockout mutant derivatives. Formation of L. rhamnosus GG biofilms on polystyrene pegs occurring after 72 h of incubation in lactobacilli AOAC medium was quantified by crystal

Techniques Used: Knock-Out, Mutagenesis, Incubation

The adhesion capacity of wild-type L. rhamnosus GG and its knockout mutant derivatives to IECs. (A) Overnight-grown L. rhamnosus GG cells were coincubated with the Caco-2 cells for 1.5 h, and thereafter the proportion of adherent bacteria, expressed as
Figure Legend Snippet: The adhesion capacity of wild-type L. rhamnosus GG and its knockout mutant derivatives to IECs. (A) Overnight-grown L. rhamnosus GG cells were coincubated with the Caco-2 cells for 1.5 h, and thereafter the proportion of adherent bacteria, expressed as

Techniques Used: Knock-Out, Mutagenesis

16) Product Images from "Cytotoxic T cell adjuvant effects of three Salmonella enterica flagellins"

Article Title: Cytotoxic T cell adjuvant effects of three Salmonella enterica flagellins

Journal: Brazilian Journal of Microbiology

doi: 10.1590/S1517-838220080001000011

Extraction of native flagellins from Salmonella enterica strains. Protein samples were sorted in polyacrylamide gels were stained with Comassie Blue (A) orcon in Western blots (B) with anti-flagellin antibodies. Samples: MW- molecular weight markers; lane 1: FliC i flagellin harvested from S . Typhimurium; lane 2: FljB flagellin harvested from S . Typhimurium; lane 3: FliC d flagellin harvested from S . Dublin SL5930 strain.
Figure Legend Snippet: Extraction of native flagellins from Salmonella enterica strains. Protein samples were sorted in polyacrylamide gels were stained with Comassie Blue (A) orcon in Western blots (B) with anti-flagellin antibodies. Samples: MW- molecular weight markers; lane 1: FliC i flagellin harvested from S . Typhimurium; lane 2: FljB flagellin harvested from S . Typhimurium; lane 3: FliC d flagellin harvested from S . Dublin SL5930 strain.

Techniques Used: Staining, Western Blot, Molecular Weight

17) Product Images from "Campylobacter jejuni Survives within Epithelial Cells by Avoiding Delivery to Lysosomes"

Article Title: Campylobacter jejuni Survives within Epithelial Cells by Avoiding Delivery to Lysosomes

Journal: PLoS Pathogens

doi: 10.1371/journal.ppat.0040014

The C. jejuni –Containing Vacuole Is Accessible to Endocytic Tracers in Macrophages but Not in Epithelial Cells (A) Cos-1 cells were infected with S. typhimurium invA (invasin) expressing the dsRed protein or C. jejuni , pulsed with the endocytic tracers dextran to label lysosomes, and phagosomes containing bacteria that colocalized with endocytic tracer were quantified. Results are the means and standard deviation of three independent experiments. For each experiment, at least 100 phagosomes were counted. Alternatively, infected cells were pulsed with the endocytic tracer BSA-gold and visualized by electron microscopy. Electron micrographs of phagosomes containing S. typhimurium invA (invasin) (B and C) or C. jejuni (D, E and F) are shown. C. jejuni –containing vacuoles (CCVs) that do not colocalize with BSA-gold (D and E) as well CCVs that colocalize with BSA-Gold and resemble lysosomes (F) are shown for comparison (see text). In addition, bone marrow–derived macrophages were infected with C. jejuni and pulsed with BSA-gold and visualized by electron microscopy. Electron micrographs of phagosomes containing C. jejuni are shown (G and H). Arrows indicate BSA-gold. Higher magnifications of the indicated insets are also shown to the right of each electron micrograph. Quantification of the colocalization of BSA-gold with bacterial vacuoles in the different cells are shown (I). Results are representative of two independent experiments in which at least 50 vacuoles were counted for colocalization with BSA-gold.
Figure Legend Snippet: The C. jejuni –Containing Vacuole Is Accessible to Endocytic Tracers in Macrophages but Not in Epithelial Cells (A) Cos-1 cells were infected with S. typhimurium invA (invasin) expressing the dsRed protein or C. jejuni , pulsed with the endocytic tracers dextran to label lysosomes, and phagosomes containing bacteria that colocalized with endocytic tracer were quantified. Results are the means and standard deviation of three independent experiments. For each experiment, at least 100 phagosomes were counted. Alternatively, infected cells were pulsed with the endocytic tracer BSA-gold and visualized by electron microscopy. Electron micrographs of phagosomes containing S. typhimurium invA (invasin) (B and C) or C. jejuni (D, E and F) are shown. C. jejuni –containing vacuoles (CCVs) that do not colocalize with BSA-gold (D and E) as well CCVs that colocalize with BSA-Gold and resemble lysosomes (F) are shown for comparison (see text). In addition, bone marrow–derived macrophages were infected with C. jejuni and pulsed with BSA-gold and visualized by electron microscopy. Electron micrographs of phagosomes containing C. jejuni are shown (G and H). Arrows indicate BSA-gold. Higher magnifications of the indicated insets are also shown to the right of each electron micrograph. Quantification of the colocalization of BSA-gold with bacterial vacuoles in the different cells are shown (I). Results are representative of two independent experiments in which at least 50 vacuoles were counted for colocalization with BSA-gold.

Techniques Used: Infection, Expressing, Standard Deviation, Electron Microscopy, Derivative Assay

The C. jejuni –Containing Vacuole Localizes in Close Proximity to the Golgi Cos-1 cells were infected with C. jejuni or S. typhimurium invA (invasin) expressing dsRed for 1 h followed by a gentamicin chase. Cells were fixed at the designated times after infection and processed for immunofluorescence using anti- C. jejuni (red) and anti-GM-130 antibodies (green). Nuclei were visualized with DAPI (blue). Immunofluorescence images of C. jejuni- infected cells 2 h (A) and 6 h (B) post-infection, and S. typhiumurium invA (invasin)-infected cells 6 h post-infection (C). (D) Electron micrograph of C. jejuni -infected cells 6 h post-infection. (E) Quantitation of C. jejuni association with the Golgi at different times after infection. Values are averages and standard deviations of three independent experiments. A minimum of 100 infected cells were counted in each experiment (F) Nocodazole treatment prevents C. jejuni 's close association with the Golgi. Cos-1 cells were infected with C. jejuni for 1 h and after a 1 h gentamicin chase, cells were treated with nocodazole for additional 4 h to disrupt microtubules. At 6 h after infection, cells were fixed and processed for immunofluorescence as described above. (G) Dymanin is required for the localization of C. jejuni in close association to the Golgi. Cos-1 cells were transfected with dynamatin p50-GFP and 24 h later, cells were infected with C. jejuni as described above. Six hours after infection, cells were fixed and processed for immunoflourecesce using anti- C. jejuni and anti GM-130 antibodies.
Figure Legend Snippet: The C. jejuni –Containing Vacuole Localizes in Close Proximity to the Golgi Cos-1 cells were infected with C. jejuni or S. typhimurium invA (invasin) expressing dsRed for 1 h followed by a gentamicin chase. Cells were fixed at the designated times after infection and processed for immunofluorescence using anti- C. jejuni (red) and anti-GM-130 antibodies (green). Nuclei were visualized with DAPI (blue). Immunofluorescence images of C. jejuni- infected cells 2 h (A) and 6 h (B) post-infection, and S. typhiumurium invA (invasin)-infected cells 6 h post-infection (C). (D) Electron micrograph of C. jejuni -infected cells 6 h post-infection. (E) Quantitation of C. jejuni association with the Golgi at different times after infection. Values are averages and standard deviations of three independent experiments. A minimum of 100 infected cells were counted in each experiment (F) Nocodazole treatment prevents C. jejuni 's close association with the Golgi. Cos-1 cells were infected with C. jejuni for 1 h and after a 1 h gentamicin chase, cells were treated with nocodazole for additional 4 h to disrupt microtubules. At 6 h after infection, cells were fixed and processed for immunofluorescence as described above. (G) Dymanin is required for the localization of C. jejuni in close association to the Golgi. Cos-1 cells were transfected with dynamatin p50-GFP and 24 h later, cells were infected with C. jejuni as described above. Six hours after infection, cells were fixed and processed for immunoflourecesce using anti- C. jejuni and anti GM-130 antibodies.

Techniques Used: Infection, Expressing, Immunofluorescence, Quantitation Assay, Transfection

The C. jejuni –Containing Vacuole Acquires Markers of the Early and Late Endocytic Pathway but Not of Lysosomes Cos-1 cells were infected with C. jejuni and or S. typhimurium invA (invasin), as indicated. At the designated times, cells were fixed and processed for immunofluorescence using antibodies directed to C. jejuni or S. typhimurium and to the endocytic markers EEA-1 and lamp-1 (A and B), or cathepsin B (G and H). Alternatively, Cos-1 cells were transfected with plasmids expressing PX-GFP (a probe for phosphoinositide 3 phosphate) (C), Rab4-GFP (D), Rab5-GFP (E), or Rab7-GFP (F) , and subsequently infected with C. jejuni or S. typhimurium invA (invasin) as indicated. Cells were fixed and processed for immunofluorescence using antibodies directed to C. jejuni or S. typhimurium . At the indicated times, the number of bacteria - containing vacuoles that colocalized with the different markers was quantified by immunofluorescence microscopy as indicated in Materials and Methods . Results are the means and standard deviation of three independent experiments. For each experiment at least 100 vacuoles were counted for acquisition of each marker.
Figure Legend Snippet: The C. jejuni –Containing Vacuole Acquires Markers of the Early and Late Endocytic Pathway but Not of Lysosomes Cos-1 cells were infected with C. jejuni and or S. typhimurium invA (invasin), as indicated. At the designated times, cells were fixed and processed for immunofluorescence using antibodies directed to C. jejuni or S. typhimurium and to the endocytic markers EEA-1 and lamp-1 (A and B), or cathepsin B (G and H). Alternatively, Cos-1 cells were transfected with plasmids expressing PX-GFP (a probe for phosphoinositide 3 phosphate) (C), Rab4-GFP (D), Rab5-GFP (E), or Rab7-GFP (F) , and subsequently infected with C. jejuni or S. typhimurium invA (invasin) as indicated. Cells were fixed and processed for immunofluorescence using antibodies directed to C. jejuni or S. typhimurium . At the indicated times, the number of bacteria - containing vacuoles that colocalized with the different markers was quantified by immunofluorescence microscopy as indicated in Materials and Methods . Results are the means and standard deviation of three independent experiments. For each experiment at least 100 vacuoles were counted for acquisition of each marker.

Techniques Used: Infection, Immunofluorescence, Transfection, Expressing, Microscopy, Standard Deviation, Marker

Dominant-Negative Rab5 and Rab7 Do Not Block Acquisition of lamp-1 by the C. jejuni –Containing Vacuole Cos-1 cells were transfected with plasmids expressing either Rab5-GFP, Rab7-GFP, or the dominant negative mutants Rab5 S34N -GFP or Rab7 N125I -GFP. Twenty-four hours after transfection, cells were infected with C. jejuni or S. typhimurium invA (invasin) for 15 min followed by a 1 h gentamicin chase. Cells were fixed, and lamp-1 acquisition was assessed after processing for immunofluorescence using anti- C. jejuni or anti- S. typhiumurium and lamp-1 antibodies. Results are the means and standard deviation of three independent experiments. For each experiment, at least 100 vacuoles were counted for colocalization with lamp-1.
Figure Legend Snippet: Dominant-Negative Rab5 and Rab7 Do Not Block Acquisition of lamp-1 by the C. jejuni –Containing Vacuole Cos-1 cells were transfected with plasmids expressing either Rab5-GFP, Rab7-GFP, or the dominant negative mutants Rab5 S34N -GFP or Rab7 N125I -GFP. Twenty-four hours after transfection, cells were infected with C. jejuni or S. typhimurium invA (invasin) for 15 min followed by a 1 h gentamicin chase. Cells were fixed, and lamp-1 acquisition was assessed after processing for immunofluorescence using anti- C. jejuni or anti- S. typhiumurium and lamp-1 antibodies. Results are the means and standard deviation of three independent experiments. For each experiment, at least 100 vacuoles were counted for colocalization with lamp-1.

Techniques Used: Dominant Negative Mutation, Blocking Assay, Transfection, Expressing, Infection, Immunofluorescence, Standard Deviation

Model for C. jejuni Internalization and Trafficking within Epithelial Cells C. jejuni enters intestinal epithelial cell via a microtubule and caveolae-dependent process. After internalization, the C. jejuni –containing vacuole transiently acquires different markers of the endocytic pathway and ultimately survives within a compartment that is functionally separated from the canonical endocytic pathway, which is represented in this scheme by S. typhimurium invA (invasin).
Figure Legend Snippet: Model for C. jejuni Internalization and Trafficking within Epithelial Cells C. jejuni enters intestinal epithelial cell via a microtubule and caveolae-dependent process. After internalization, the C. jejuni –containing vacuole transiently acquires different markers of the endocytic pathway and ultimately survives within a compartment that is functionally separated from the canonical endocytic pathway, which is represented in this scheme by S. typhimurium invA (invasin).

Techniques Used:

18) Product Images from "In Salmonella enterica, OatA (Formerly YjgM) Uses O-Acetyl-Serine and Acetyl-CoA to Synthesize N,O-Diacetylserine, Which Upregulates Cysteine Biosynthesis"

Article Title: In Salmonella enterica, OatA (Formerly YjgM) Uses O-Acetyl-Serine and Acetyl-CoA to Synthesize N,O-Diacetylserine, Which Upregulates Cysteine Biosynthesis

Journal: Frontiers in Microbiology

doi: 10.3389/fmicb.2018.02838

Acetyl-serine analogs: NAS, OAS, and DAS. NAS (left) is thought to be a ligand for CysB, which is produced from non-enzymatic O -to- N intramolecular migration of OAS (center) at ≥pH 7.6 at a rate of 1% min -1 . DAS (right) is produced from acetylation of OAS by the S. enterica OatA N -acetyltransferase.
Figure Legend Snippet: Acetyl-serine analogs: NAS, OAS, and DAS. NAS (left) is thought to be a ligand for CysB, which is produced from non-enzymatic O -to- N intramolecular migration of OAS (center) at ≥pH 7.6 at a rate of 1% min -1 . DAS (right) is produced from acetylation of OAS by the S. enterica OatA N -acetyltransferase.

Techniques Used: Produced, Migration

Pathway for biosynthesis of L -Cys in Salmonella enterica. Sulfate is sequestered by the sulfate-binding protein (sbp) and uptake is mediated by the CysUWA sulfate/thiosulfate transporter. The ATP sulfurylase [CysD (catalytic subunit)/CysN (GTP-binding subunit), EC 2.7.7.4] activates sulfate to adenosine 5′-phosphate (APS), which is a substrate for APS kinase (CysC, EC 2.7.1.25). 3-phosphoadenosine 5′-phosphosulfate (PAPS) is reduced to sulfite by the PAPS reductase (CysH, EC 1.8.4.8) and sulfite is reduced to sulfide by a NADPH-sulfite reductase (CysJI, EC 1.8.1.2). A serine acetyltransferase (CysE, EC 2.3.1.30) produces OAS from L -Ser and AcCoA. OAS and sulfide are then substrates for cysteine synthase (CysK, EC 2.5.1.47), which produces L -Cys. Transcription of cysJIH and cysK is upregulated upon binding of CysB and acetyl-serine.
Figure Legend Snippet: Pathway for biosynthesis of L -Cys in Salmonella enterica. Sulfate is sequestered by the sulfate-binding protein (sbp) and uptake is mediated by the CysUWA sulfate/thiosulfate transporter. The ATP sulfurylase [CysD (catalytic subunit)/CysN (GTP-binding subunit), EC 2.7.7.4] activates sulfate to adenosine 5′-phosphate (APS), which is a substrate for APS kinase (CysC, EC 2.7.1.25). 3-phosphoadenosine 5′-phosphosulfate (PAPS) is reduced to sulfite by the PAPS reductase (CysH, EC 1.8.4.8) and sulfite is reduced to sulfide by a NADPH-sulfite reductase (CysJI, EC 1.8.1.2). A serine acetyltransferase (CysE, EC 2.3.1.30) produces OAS from L -Ser and AcCoA. OAS and sulfide are then substrates for cysteine synthase (CysK, EC 2.5.1.47), which produces L -Cys. Transcription of cysJIH and cysK is upregulated upon binding of CysB and acetyl-serine.

Techniques Used: Binding Assay, Papanicolaou Stain

Effects of oatA overexpression on transcription of cysteine regulon genes. Total RNA was extracted from cells grown on glycerol (22 mM) minimal medium with MgCl 2 (1 mM) in lieu of MgSO 4 . Cells were harvested at mid-log phase, where oatA overexpression caused a growth phenotype, as shown in Figure 9 . Expression of cysB (A) , cysD (B) , cysI (C) , cysK (D) , and cysM (E) was assessed using RT-qPCR and transcripts were normalized to the gyrB gene of S. enterica . Changes in transcription when oatA was overexpressed (induction with 1 mM L -arabinose) were observed cysB (fivefold), cysK (fivefold), cysD (threefold), and cysI (threefold). Transcription was not affected with relation to cysM . Experiment was completed in three independent experiments, with RNA harvested in biological triplicate for each experiment and RT-qPCR completed in technical triplicate for each RNA replicate. Values shown are averages of biological triplicates with error bars representing standard deviation. P -values were calculated and represented by ∗∗
Figure Legend Snippet: Effects of oatA overexpression on transcription of cysteine regulon genes. Total RNA was extracted from cells grown on glycerol (22 mM) minimal medium with MgCl 2 (1 mM) in lieu of MgSO 4 . Cells were harvested at mid-log phase, where oatA overexpression caused a growth phenotype, as shown in Figure 9 . Expression of cysB (A) , cysD (B) , cysI (C) , cysK (D) , and cysM (E) was assessed using RT-qPCR and transcripts were normalized to the gyrB gene of S. enterica . Changes in transcription when oatA was overexpressed (induction with 1 mM L -arabinose) were observed cysB (fivefold), cysK (fivefold), cysD (threefold), and cysI (threefold). Transcription was not affected with relation to cysM . Experiment was completed in three independent experiments, with RNA harvested in biological triplicate for each experiment and RT-qPCR completed in technical triplicate for each RNA replicate. Values shown are averages of biological triplicates with error bars representing standard deviation. P -values were calculated and represented by ∗∗

Techniques Used: Over Expression, Expressing, Quantitative RT-PCR, Standard Deviation

19) Product Images from "Effect of NaCl on Heat Resistance, Antibiotic Susceptibility, and Caco-2 Cell Invasion of Salmonella"

Article Title: Effect of NaCl on Heat Resistance, Antibiotic Susceptibility, and Caco-2 Cell Invasion of Salmonella

Journal: BioMed Research International

doi: 10.1155/2013/274096

Invasion efficiency of S. typhimurium NCCP10812 and S. enteritidis NCCP12243 exposed to single concentrations (0, 2, and 4%) of NaCl (a) and sequentially increased NaCl concentrations up to 4% (b). a–d Means with different superscript letters are different ( P
Figure Legend Snippet: Invasion efficiency of S. typhimurium NCCP10812 and S. enteritidis NCCP12243 exposed to single concentrations (0, 2, and 4%) of NaCl (a) and sequentially increased NaCl concentrations up to 4% (b). a–d Means with different superscript letters are different ( P

Techniques Used:

20) Product Images from "Evaluation of Near-Infrared Pasteurization in Controlling Escherichia coli O157:H7, Salmonella enterica Serovar Typhimurium, and Listeria monocytogenes in Ready-To-Eat Sliced Ham"

Article Title: Evaluation of Near-Infrared Pasteurization in Controlling Escherichia coli O157:H7, Salmonella enterica Serovar Typhimurium, and Listeria monocytogenes in Ready-To-Eat Sliced Ham

Journal: Applied and Environmental Microbiology

doi: 10.1128/AEM.00942-12

Survival curves of Salmonella Typhimurium, Escherichia coli O157:H7, and Listeria monocytogenes inside two contiguous ham slices treated with NIR or conventional convective heating. The error bars indicate standard deviations calculated from triplicates.
Figure Legend Snippet: Survival curves of Salmonella Typhimurium, Escherichia coli O157:H7, and Listeria monocytogenes inside two contiguous ham slices treated with NIR or conventional convective heating. The error bars indicate standard deviations calculated from triplicates.

Techniques Used:

Survival curves of Salmonella Typhimurium, Escherichia coli O157:H7, and Listeria monocytogenes on ham slice surfaces treated with NIR or conventional convective heating. The error bars indicate standard deviations calculated from triplicates.
Figure Legend Snippet: Survival curves of Salmonella Typhimurium, Escherichia coli O157:H7, and Listeria monocytogenes on ham slice surfaces treated with NIR or conventional convective heating. The error bars indicate standard deviations calculated from triplicates.

Techniques Used:

21) Product Images from "Uropathogenic Escherichia coli Invades Host Cells via an HDAC6-modulated Microtubule-dependent Pathway * Invades Host Cells via an HDAC6-modulated Microtubule-dependent Pathway * S⃞"

Article Title: Uropathogenic Escherichia coli Invades Host Cells via an HDAC6-modulated Microtubule-dependent Pathway * Invades Host Cells via an HDAC6-modulated Microtubule-dependent Pathway * S⃞

Journal: The Journal of Biological Chemistry

doi: 10.1074/jbc.M805010200

Aurora A kinase-dependent invasion of host cells by UPEC. a and b , 5637 bladder cells were pretreated for 3 h with 10 μ m of AKI II or carrier alone. Cells were then infected with UTI89 for 2 h in the continued presence of drug or carrier after
Figure Legend Snippet: Aurora A kinase-dependent invasion of host cells by UPEC. a and b , 5637 bladder cells were pretreated for 3 h with 10 μ m of AKI II or carrier alone. Cells were then infected with UTI89 for 2 h in the continued presence of drug or carrier after

Techniques Used: Infection

Microtubule-dependent invasion of bladder cells by UPEC. 5637 bladder cell monolayers were treated with the indicated concentrations of ( a ) nocodazole, ( b ) taxol, or ( c ) vinblastine for 1 h prior to infection with the UPEC isolate UTI89. After a 2-h
Figure Legend Snippet: Microtubule-dependent invasion of bladder cells by UPEC. 5637 bladder cell monolayers were treated with the indicated concentrations of ( a ) nocodazole, ( b ) taxol, or ( c ) vinblastine for 1 h prior to infection with the UPEC isolate UTI89. After a 2-h

Techniques Used: Infection

Kinesin-1 promotes host cell invasion by UPEC. Bladder cells were pre-treated with aurintricarboxylic acid (ATA, 50 μ m ) for 3 h or with sodium orthovanadate (Na 3 VO 4 , 100 μ m ) for 1 h prior to infection with UTI89. After 2 h in the continued
Figure Legend Snippet: Kinesin-1 promotes host cell invasion by UPEC. Bladder cells were pre-treated with aurintricarboxylic acid (ATA, 50 μ m ) for 3 h or with sodium orthovanadate (Na 3 VO 4 , 100 μ m ) for 1 h prior to infection with UTI89. After 2 h in the continued

Techniques Used: Infection

22) Product Images from "A novel CsrA titration mechanism regulates fimbrial gene expression in Salmonella typhimurium"

Article Title: A novel CsrA titration mechanism regulates fimbrial gene expression in Salmonella typhimurium

Journal: The EMBO Journal

doi: 10.1038/emboj.2013.206

CsrA regulates expression of PefA. ( A ) Expression of FimA was detected in cell lysates of the indicated S. typhimurium strains using western blot. ( B ) Quantification of FimA levels in western blots ( N =3) by densitometry. ( C ) Relative expression of fimA
Figure Legend Snippet: CsrA regulates expression of PefA. ( A ) Expression of FimA was detected in cell lysates of the indicated S. typhimurium strains using western blot. ( B ) Quantification of FimA levels in western blots ( N =3) by densitometry. ( C ) Relative expression of fimA

Techniques Used: Expressing, Western Blot

SirA represses expression of PefA via downregulation of CsrB and CsrC. Expression of CsrB RNA ( A ) and CsrC RNA ( B ) was quantified by real-time PCR using RNA isolated from S. typhimurium wild type (wt) and the sirA mutant ( sirA ). Bars represent the average
Figure Legend Snippet: SirA represses expression of PefA via downregulation of CsrB and CsrC. Expression of CsrB RNA ( A ) and CsrC RNA ( B ) was quantified by real-time PCR using RNA isolated from S. typhimurium wild type (wt) and the sirA mutant ( sirA ). Bars represent the average

Techniques Used: Expressing, Real-time Polymerase Chain Reaction, Isolation, Mutagenesis

The CsrA-binding site in the 5′-UTR of the pefA transcript limits expression to host environments. Streptomycin pretreated mice were inoculated with the indicated S. typhimurium mutants and RNA was isolated from the inoculum cultures (inoculum)
Figure Legend Snippet: The CsrA-binding site in the 5′-UTR of the pefA transcript limits expression to host environments. Streptomycin pretreated mice were inoculated with the indicated S. typhimurium mutants and RNA was isolated from the inoculum cultures (inoculum)

Techniques Used: Binding Assay, Expressing, Mouse Assay, Isolation

Proposed model for the hierarchical control of fimbrial expression in S. typhimurium mediated by CsrA. The exceptionally high transcript levels of the fimAICDHF 5′-UTR can antagonize the regulatory effects of CsrA by binding and sequestering this
Figure Legend Snippet: Proposed model for the hierarchical control of fimbrial expression in S. typhimurium mediated by CsrA. The exceptionally high transcript levels of the fimAICDHF 5′-UTR can antagonize the regulatory effects of CsrA by binding and sequestering this

Techniques Used: Expressing, Binding Assay

23) Product Images from "Genetics of Swarming Motility in Salmonella enterica Serovar Typhimurium: Critical Role for Lipopolysaccharide"

Article Title: Genetics of Swarming Motility in Salmonella enterica Serovar Typhimurium: Critical Role for Lipopolysaccharide

Journal: Journal of Bacteriology

doi:

Rescue of swarming defects with external addition of surfactin and LPS. Five-microliter drops of solution containing surfactin or LPS were placed at the center of 0.5% Difco swarm medium, inoculated with S. enterica serovar Typhimurium ( waaL and cheA ) and E. coli (RP437) strains, and incubated overnight as described in Materials and Methods.
Figure Legend Snippet: Rescue of swarming defects with external addition of surfactin and LPS. Five-microliter drops of solution containing surfactin or LPS were placed at the center of 0.5% Difco swarm medium, inoculated with S. enterica serovar Typhimurium ( waaL and cheA ) and E. coli (RP437) strains, and incubated overnight as described in Materials and Methods.

Techniques Used: Incubation

Precocious swarming of waaG LPS core mutant. (A) A wild-type (wt) S. enterica serovar Typhimurium strain (SL3770) and its waaG mutant derivative (SL3769) were inoculated on Difco Bacto swarm medium and incubated at 37°C. Photographs were taken after 5 and 20 h of incubation. The arrow points to edge of the waaG swarm colony. (B) The waaG strain was inoculated on minimal swarm medium and incubated at 37°C. Photographs were taken after 24, 30, and 44 h. The arrow indicates edge of the slime ring. Swarming movement was confined to regions that appear as branches emanating from the center.
Figure Legend Snippet: Precocious swarming of waaG LPS core mutant. (A) A wild-type (wt) S. enterica serovar Typhimurium strain (SL3770) and its waaG mutant derivative (SL3769) were inoculated on Difco Bacto swarm medium and incubated at 37°C. Photographs were taken after 5 and 20 h of incubation. The arrow points to edge of the waaG swarm colony. (B) The waaG strain was inoculated on minimal swarm medium and incubated at 37°C. Photographs were taken after 24, 30, and 44 h. The arrow indicates edge of the slime ring. Swarming movement was confined to regions that appear as branches emanating from the center.

Techniques Used: Mutagenesis, Incubation

24) Product Images from "InvF Is Required for Expression of Genes Encoding Proteins Secreted by the SPI1 Type III Secretion Apparatus in Salmonella typhimurium"

Article Title: InvF Is Required for Expression of Genes Encoding Proteins Secreted by the SPI1 Type III Secretion Apparatus in Salmonella typhimurium

Journal: Journal of Bacteriology

doi:

Supernatant proteins from wild-type and Δ invF strains. Lane 1, supernatant proteins from the secretion defective spaS mutant SVM514; lanes 2 to 7, S. typhimurium SL1344 containing pWSK130, pHD9 ( invF + ), and p hilA (lanes 2 to 4) and the invF mutant SVM579 containing the same plasmids in the same order (lanes 5 to 7); lanes 8 and 9, supernatant proteins from SVM579 strains containing the vector pVLT33 (lane 8) and the IPTG-inducible sigDE clone pHH37 (lane 9). Positions of molecular weight standards are indicated in kilodaltons on the left; previously identified secreted proteins are indicated on the right. Question marks denote proteins that have not been confirmed by immunoblot analysis. Proteins were prepared and analyzed as described in Materials and Methods.
Figure Legend Snippet: Supernatant proteins from wild-type and Δ invF strains. Lane 1, supernatant proteins from the secretion defective spaS mutant SVM514; lanes 2 to 7, S. typhimurium SL1344 containing pWSK130, pHD9 ( invF + ), and p hilA (lanes 2 to 4) and the invF mutant SVM579 containing the same plasmids in the same order (lanes 5 to 7); lanes 8 and 9, supernatant proteins from SVM579 strains containing the vector pVLT33 (lane 8) and the IPTG-inducible sigDE clone pHH37 (lane 9). Positions of molecular weight standards are indicated in kilodaltons on the left; previously identified secreted proteins are indicated on the right. Question marks denote proteins that have not been confirmed by immunoblot analysis. Proteins were prepared and analyzed as described in Materials and Methods.

Techniques Used: Mutagenesis, Plasmid Preparation, Molecular Weight

Model for the regulation of invasion/virulence gene expression in S. typhimurium . The direction of transcription for each gene cluster is indicated by closed arrows; open arrows represent putative transcripts of the inv-spa and sip/ssp genes. Question marks indicate either unidentified regulatory factors or unclear relationships between the designated regulator and the noted promoter.
Figure Legend Snippet: Model for the regulation of invasion/virulence gene expression in S. typhimurium . The direction of transcription for each gene cluster is indicated by closed arrows; open arrows represent putative transcripts of the inv-spa and sip/ssp genes. Question marks indicate either unidentified regulatory factors or unclear relationships between the designated regulator and the noted promoter.

Techniques Used: Expressing

25) Product Images from "Decontamination Effect of the Spindle and 222-Nanometer Krypton-Chlorine Excimer Lamp Combination against Pathogens on Apples (Malus domestica Borkh.) and Bell Peppers (Capsicum annuum L.)"

Article Title: Decontamination Effect of the Spindle and 222-Nanometer Krypton-Chlorine Excimer Lamp Combination against Pathogens on Apples (Malus domestica Borkh.) and Bell Peppers (Capsicum annuum L.)

Journal: Applied and Environmental Microbiology

doi: 10.1128/AEM.00006-19

Surviving populations (log CFU/sample or log CFU/250 ml) of Escherichia coli O157:H7, Salmonella Typhimurium, and Listeria monocytogenes on sample surfaces or in washing solution (tap water [TW] plus 0.1% Tween 20) treated with the Spindle in
Figure Legend Snippet: Surviving populations (log CFU/sample or log CFU/250 ml) of Escherichia coli O157:H7, Salmonella Typhimurium, and Listeria monocytogenes on sample surfaces or in washing solution (tap water [TW] plus 0.1% Tween 20) treated with the Spindle in

Techniques Used:

Surviving populations (log CFU/sample or log CFU/250 ml) of Escherichia coli O157:H7, Salmonella Typhimurium, and Listeria monocytogenes on the sample surface or in washing solution (tap water [TW]) treated with the Spindle in combination with
Figure Legend Snippet: Surviving populations (log CFU/sample or log CFU/250 ml) of Escherichia coli O157:H7, Salmonella Typhimurium, and Listeria monocytogenes on the sample surface or in washing solution (tap water [TW]) treated with the Spindle in combination with

Techniques Used:

26) Product Images from "Evaluation of Sanitizing Methods for Reducing Microbial Contamination on Fresh Strawberry, Cherry Tomato, and Red Bayberry"

Article Title: Evaluation of Sanitizing Methods for Reducing Microbial Contamination on Fresh Strawberry, Cherry Tomato, and Red Bayberry

Journal: Frontiers in Microbiology

doi: 10.3389/fmicb.2017.02397

Reductions in aerobic bacteria (A) , E. coli (B) , mold (C) , yeast (D) , and S. Typhimurium (E) on cherry tomato achieved by different sanitizing methods. Data are expressed as mean ± SD ( n = 3). ∗∗ P
Figure Legend Snippet: Reductions in aerobic bacteria (A) , E. coli (B) , mold (C) , yeast (D) , and S. Typhimurium (E) on cherry tomato achieved by different sanitizing methods. Data are expressed as mean ± SD ( n = 3). ∗∗ P

Techniques Used:

Reductions in aerobic bacteria (A) , E. coli (B) , mold (C) , yeast (D) , and S. Typhimurium (E) on strawberry achieved by different sanitizing methods. Data are expressed as mean ± SD ( n = 3). ∗ P
Figure Legend Snippet: Reductions in aerobic bacteria (A) , E. coli (B) , mold (C) , yeast (D) , and S. Typhimurium (E) on strawberry achieved by different sanitizing methods. Data are expressed as mean ± SD ( n = 3). ∗ P

Techniques Used:

27) Product Images from "Commensal Akkermansia muciniphila Exacerbates Gut Inflammation in Salmonella Typhimurium-Infected Gnotobiotic Mice"

Article Title: Commensal Akkermansia muciniphila Exacerbates Gut Inflammation in Salmonella Typhimurium-Infected Gnotobiotic Mice

Journal: PLoS ONE

doi: 10.1371/journal.pone.0074963

SIHUMI mice colonized with both A. muciniphila and S. Typhimurium display an increased cecal macrophage infiltration. (A) Formalin fixed paraffin embedded cecum tissue was thin sectioned at 2 µm. Macrophages were stained by targeting the F4/80 receptor expressed on mouse macrophages using immunohistochemistry with specific antibodies. Brown color indicates positively stained macrophages. Magnification 400-fold. Bar = 100 µm. (B) Positively stained macrophages were enumerated along a stretch of 50 µm of lamina muscularis for both lamina propria and sub-mucosa (see materials and methods). SIHUMI mice colonized with both A. muciniphila and S. Typhimurium had the highest macrophage infiltration scores compared to the other groups (see Figure. 1). Data are expressed as median with range. n = 5 mice per group. *P
Figure Legend Snippet: SIHUMI mice colonized with both A. muciniphila and S. Typhimurium display an increased cecal macrophage infiltration. (A) Formalin fixed paraffin embedded cecum tissue was thin sectioned at 2 µm. Macrophages were stained by targeting the F4/80 receptor expressed on mouse macrophages using immunohistochemistry with specific antibodies. Brown color indicates positively stained macrophages. Magnification 400-fold. Bar = 100 µm. (B) Positively stained macrophages were enumerated along a stretch of 50 µm of lamina muscularis for both lamina propria and sub-mucosa (see materials and methods). SIHUMI mice colonized with both A. muciniphila and S. Typhimurium had the highest macrophage infiltration scores compared to the other groups (see Figure. 1). Data are expressed as median with range. n = 5 mice per group. *P

Techniques Used: Mouse Assay, Formalin-fixed Paraffin-Embedded, Staining, Immunohistochemistry

Concomitant presence of A. muciniphila and S. Typhimurium results in increased histopathology scores in SIHUMI mice. (A) Gnotobiotic C3H mice containing 8 defined microbial species (SIHUMI) were subsequently inoculated with A. muciniphila or S. Typhimurium or consecutively with both organisms (see Figure 1 ). SIHUMI and SIHUMI-A mice had the lowest histopathology scores (≤4.0) with no signs of inflammation and were therefore taken as baseline (dotted line). Data are expressed as median with range. *P
Figure Legend Snippet: Concomitant presence of A. muciniphila and S. Typhimurium results in increased histopathology scores in SIHUMI mice. (A) Gnotobiotic C3H mice containing 8 defined microbial species (SIHUMI) were subsequently inoculated with A. muciniphila or S. Typhimurium or consecutively with both organisms (see Figure 1 ). SIHUMI and SIHUMI-A mice had the lowest histopathology scores (≤4.0) with no signs of inflammation and were therefore taken as baseline (dotted line). Data are expressed as median with range. *P

Techniques Used: Histopathology, Mouse Assay

SIHUMI mice colonized with both A. muciniphila and S. Typhimurium display enlarged mLN and elevated S. Typhimurium cell numbers. (A) Mesenteric lymph nodes (mLN) were obtained from four groups of gnotobiotic C3H mice. SIHUMI mice were subsequently inoculated with A. muciniphila or S. Typhimurium or consecutively with both organisms (see Figure 1 ). The mLN tissue was homogenized and DNA was isolated to quantify S. Typhimurium using quantitative PCR with primers targeting the ttr-region of S. Typhimurium. Absolute cell numbers were calculated based on calibration curves with known concentrations of S. Typhimurium. The mLN of SIHUMI-AS mice contained 10-fold higher cell numbers of S. Typhimurium compared to SIHUMI-S mice. Data are expressed as mean±standard error. n = 10 mice per group. *P
Figure Legend Snippet: SIHUMI mice colonized with both A. muciniphila and S. Typhimurium display enlarged mLN and elevated S. Typhimurium cell numbers. (A) Mesenteric lymph nodes (mLN) were obtained from four groups of gnotobiotic C3H mice. SIHUMI mice were subsequently inoculated with A. muciniphila or S. Typhimurium or consecutively with both organisms (see Figure 1 ). The mLN tissue was homogenized and DNA was isolated to quantify S. Typhimurium using quantitative PCR with primers targeting the ttr-region of S. Typhimurium. Absolute cell numbers were calculated based on calibration curves with known concentrations of S. Typhimurium. The mLN of SIHUMI-AS mice contained 10-fold higher cell numbers of S. Typhimurium compared to SIHUMI-S mice. Data are expressed as mean±standard error. n = 10 mice per group. *P

Techniques Used: Mouse Assay, Isolation, Real-time Polymerase Chain Reaction

SIHUMI mice colonized with both A. muciniphila and S. Typhimurium display reduced mucus sulphation. Formalin fixed thin sections (4 µm) of cecal tissue of mice belonging to either one of four groups: SIHUMI, SIHUMI-A, SIHUMI-S and SIHUMI-AS (see Figure. 1) were stained with high iron diamine (HID)/AB at pH-2.5 and subsequently analyzed. Brown color indicates sulphated mucins while blue color indicates sialylated mucins. SIHUMI-AS mice display few sulphated mucins compared to the other mouse groups. Magnification 400×. Bars indicate 100 µm.
Figure Legend Snippet: SIHUMI mice colonized with both A. muciniphila and S. Typhimurium display reduced mucus sulphation. Formalin fixed thin sections (4 µm) of cecal tissue of mice belonging to either one of four groups: SIHUMI, SIHUMI-A, SIHUMI-S and SIHUMI-AS (see Figure. 1) were stained with high iron diamine (HID)/AB at pH-2.5 and subsequently analyzed. Brown color indicates sulphated mucins while blue color indicates sialylated mucins. SIHUMI-AS mice display few sulphated mucins compared to the other mouse groups. Magnification 400×. Bars indicate 100 µm.

Techniques Used: Mouse Assay, Staining

Hypothetical Scheme. The presence of A. muciniphila, leads to the exacerbation of S. Typhimurium-induced intestinal inflammation. We propose that the presence of A. muciniphila causes changes in mucin composition and production, which in turn facilitates the invasion of S. Typhimurium into the host. Increased inflammatory status was characterized by increased pro-inflammatory cytokines, increased macrophage infiltration and invasion of the pathogen into the lymph nodes, reduced number of mucin-filled goblet cells in SIHUMI-AS mice (B) compared to SIHUMI-S mice (A). Our data suggests that in the presence of both A. muciniphila and S. Typhimurium, mucus sulphation is diminished and this may facilitate the access of S. Typhimurium to sialic acid in mucus. Sialic acid may serve as a substrate and adhesion site for S. Typhimurium in the gut [56] , [57] . Increased gene expression of IFN-γ and IP-10 indicate an increased NK-cell recruitment. mLN - mesenteric lymph nodes, NK- Natural killer cells. (↑ increased; ↓ decreased; grey dotted line: assumed processes including lectin-sialic acid binding [56] , M-cells for pathogen transit [43] , [58] , [59] ; black line: supported by data of the present study).
Figure Legend Snippet: Hypothetical Scheme. The presence of A. muciniphila, leads to the exacerbation of S. Typhimurium-induced intestinal inflammation. We propose that the presence of A. muciniphila causes changes in mucin composition and production, which in turn facilitates the invasion of S. Typhimurium into the host. Increased inflammatory status was characterized by increased pro-inflammatory cytokines, increased macrophage infiltration and invasion of the pathogen into the lymph nodes, reduced number of mucin-filled goblet cells in SIHUMI-AS mice (B) compared to SIHUMI-S mice (A). Our data suggests that in the presence of both A. muciniphila and S. Typhimurium, mucus sulphation is diminished and this may facilitate the access of S. Typhimurium to sialic acid in mucus. Sialic acid may serve as a substrate and adhesion site for S. Typhimurium in the gut [56] , [57] . Increased gene expression of IFN-γ and IP-10 indicate an increased NK-cell recruitment. mLN - mesenteric lymph nodes, NK- Natural killer cells. (↑ increased; ↓ decreased; grey dotted line: assumed processes including lectin-sialic acid binding [56] , M-cells for pathogen transit [43] , [58] , [59] ; black line: supported by data of the present study).

Techniques Used: Mouse Assay, Expressing, Binding Assay

Presence of A. muciniphila renders S. Typhimurium the dominant species in gnotobiotic SIHUMI mice. Cecal contents were collected from gnotobiotic C3H mice, differing in their microbial status: (A) Mice with a defined microbial community of eight bacterial species (SIHUMI), (B) SIHUMI mice additionally colonized with A. muciniphila (SIHUMI-A), (C) SIHUMI mice infected with S. Typhimurium (SIHUMI-S) and (D) SIHUMI mice colonized with A. muciniphila and 10 days later infected with S. Typhimurium (SIHUMI-AS) (see Figure 1 ). Total DNA was extracted and bacterial cell numbers were quantified by qPCR with primers targeting the HSP60 gene of the SIHUMI members, the 16S rRNA gene of A. muciniphila and the ttr-region of S. Typhimurium. Calculation of the cell numbers was based on DNA obtained from cell suspensions containing known cell numbers of the targeted bacterial species (see materials and methods). Presence of A. muciniphila in SIHUMI-AS mice is attributed to an increase in the proportion of S. Typhimurium cells at the expense of other community members showing reduced proportion of SIHUMI members. Ten animals per group were used. The bacterial cell numbers and P-values for the differences between the groups are provided in Table 1 .
Figure Legend Snippet: Presence of A. muciniphila renders S. Typhimurium the dominant species in gnotobiotic SIHUMI mice. Cecal contents were collected from gnotobiotic C3H mice, differing in their microbial status: (A) Mice with a defined microbial community of eight bacterial species (SIHUMI), (B) SIHUMI mice additionally colonized with A. muciniphila (SIHUMI-A), (C) SIHUMI mice infected with S. Typhimurium (SIHUMI-S) and (D) SIHUMI mice colonized with A. muciniphila and 10 days later infected with S. Typhimurium (SIHUMI-AS) (see Figure 1 ). Total DNA was extracted and bacterial cell numbers were quantified by qPCR with primers targeting the HSP60 gene of the SIHUMI members, the 16S rRNA gene of A. muciniphila and the ttr-region of S. Typhimurium. Calculation of the cell numbers was based on DNA obtained from cell suspensions containing known cell numbers of the targeted bacterial species (see materials and methods). Presence of A. muciniphila in SIHUMI-AS mice is attributed to an increase in the proportion of S. Typhimurium cells at the expense of other community members showing reduced proportion of SIHUMI members. Ten animals per group were used. The bacterial cell numbers and P-values for the differences between the groups are provided in Table 1 .

Techniques Used: Mouse Assay, Infection, Real-time Polymerase Chain Reaction

Presence of both A. muciniphila and S. Typhimurium is accompanied by increased pro-inflammatory cytokines. (A) Cecal mRNA levels of IFN-γ, IP-10, TNF-α, IL-12, IL-6, IL-17 and IL-18 in gnotobiotic SIHUMI mice were measured. mRNA was extracted from cecum mucosa of mice belonging to either one of four groups: SIHUMI, SIHUMI-A, SIHUMI-S and SIHUMI-AS (see Figure. 1). The mRNA was converted to cDNA for quantitative real-time PCR measurement (see materials and methods). Inoculation of the gnotobiotic SIHUMI mice with A. muciniphila followed by S. Typhimurium infection (SIHUMI-AS) caused an increase in mRNA levels of pro-inflammatory cytokines except IL-18. Data are expressed as mean±standard error. n = 6 per group. Star indicates statistically significant differences ( *P
Figure Legend Snippet: Presence of both A. muciniphila and S. Typhimurium is accompanied by increased pro-inflammatory cytokines. (A) Cecal mRNA levels of IFN-γ, IP-10, TNF-α, IL-12, IL-6, IL-17 and IL-18 in gnotobiotic SIHUMI mice were measured. mRNA was extracted from cecum mucosa of mice belonging to either one of four groups: SIHUMI, SIHUMI-A, SIHUMI-S and SIHUMI-AS (see Figure. 1). The mRNA was converted to cDNA for quantitative real-time PCR measurement (see materials and methods). Inoculation of the gnotobiotic SIHUMI mice with A. muciniphila followed by S. Typhimurium infection (SIHUMI-AS) caused an increase in mRNA levels of pro-inflammatory cytokines except IL-18. Data are expressed as mean±standard error. n = 6 per group. Star indicates statistically significant differences ( *P

Techniques Used: Mouse Assay, Real-time Polymerase Chain Reaction, Infection

SIHUMI mice with both A. muciniphila and S. Typhimurium display increased MUC2 mRNA levels (A) and reduced numbers of mucin filled goblet cells (B and C). (A) mRNA was extracted from cecum mucosa of mice belonging to either one of four groups: SIHUMI, SIHUMI-A, SIHUMI-S and SIHUMI-AS. MUC2 mRNA from cecum mucosa was converted to cDNA and expression levels were quantified using real-time PCR (see materials and methods). SIHUMI-A and SIHUMI-AS mice showed significantly higher MUC2 gene expression compared to the other two groups, harboring no A. muciniphila . Data are expressed as mean±standard error. n = 6 per group. *P
Figure Legend Snippet: SIHUMI mice with both A. muciniphila and S. Typhimurium display increased MUC2 mRNA levels (A) and reduced numbers of mucin filled goblet cells (B and C). (A) mRNA was extracted from cecum mucosa of mice belonging to either one of four groups: SIHUMI, SIHUMI-A, SIHUMI-S and SIHUMI-AS. MUC2 mRNA from cecum mucosa was converted to cDNA and expression levels were quantified using real-time PCR (see materials and methods). SIHUMI-A and SIHUMI-AS mice showed significantly higher MUC2 gene expression compared to the other two groups, harboring no A. muciniphila . Data are expressed as mean±standard error. n = 6 per group. *P

Techniques Used: Mouse Assay, Expressing, Real-time Polymerase Chain Reaction

Design of the animal experiment. Fourty C3H mice associated with a defined microbial community of 8 bacterial species (SIHUMI) were allocated to four different groups (10 mice per group). Each mouse was associated with 8 bacterial species (SIHUMI). Twelve weeks-old SIHUMI mice were subsequently associated with A. muciniphila (SIHUMI-A) or S. Typhimurium (SIHUMI-S) or with both A. muciniphila and S. Typhimurium (SIHUMI-AS). SIHUMI mice received only sterile medium. Times of association, infection and killing are as indicated. ‡ - killed.
Figure Legend Snippet: Design of the animal experiment. Fourty C3H mice associated with a defined microbial community of 8 bacterial species (SIHUMI) were allocated to four different groups (10 mice per group). Each mouse was associated with 8 bacterial species (SIHUMI). Twelve weeks-old SIHUMI mice were subsequently associated with A. muciniphila (SIHUMI-A) or S. Typhimurium (SIHUMI-S) or with both A. muciniphila and S. Typhimurium (SIHUMI-AS). SIHUMI mice received only sterile medium. Times of association, infection and killing are as indicated. ‡ - killed.

Techniques Used: Mouse Assay, Infection

28) Product Images from "In Vitro and In Vivo Antibacterial Activity of Punica granatum Peel Ethanol Extract against Salmonella"

Article Title: In Vitro and In Vivo Antibacterial Activity of Punica granatum Peel Ethanol Extract against Salmonella

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

doi: 10.1093/ecam/nep105

Importance of PGPE against S. typhimurium infection.
Figure Legend Snippet: Importance of PGPE against S. typhimurium infection.

Techniques Used: Infection

29) Product Images from "Structure-function analyses involving palindromic analogs of tritrypticin suggest autonomy of anti-endotoxin and antibacterial activities"

Article Title: Structure-function analyses involving palindromic analogs of tritrypticin suggest autonomy of anti-endotoxin and antibacterial activities

Journal: Protein Science : A Publication of the Protein Society

doi: 10.1110/ps.073145008

Comparison of NT7 and CT7 for LPS binding by competitive displacement of dansyl polymyxin B in a dose-dependent manner.
Figure Legend Snippet: Comparison of NT7 and CT7 for LPS binding by competitive displacement of dansyl polymyxin B in a dose-dependent manner.

Techniques Used: Binding Assay

Comparison of circular dichroism profiles of ( A ) NT7 and ( B ) CT7, in free and LPS-bound forms.
Figure Legend Snippet: Comparison of circular dichroism profiles of ( A ) NT7 and ( B ) CT7, in free and LPS-bound forms.

Techniques Used:

30) Product Images from "Antioxidant oils and Salmonella enterica Typhimurium reduce tumor in an experimental model of hepatic metastasis"

Article Title: Antioxidant oils and Salmonella enterica Typhimurium reduce tumor in an experimental model of hepatic metastasis

Journal: OncoTargets and therapy

doi: 10.2147/OTT.S17081

Effect of combination antioxidant seed oil on the splenic lymphocyte response to SalpIL2 in mice. Splenic natural killer (NK) ( A ), CD 8 + T ( B ), and CD 4 + T cell populations ( C ) as determined by flow cytometry in response in animals fed a diet consisting of equal amounts of black raspberry (BR; Rubus occidentalis ), black cumin (BC; Nigella sativa ) seed oils (○), administered a single oral dose of SalpIL2 (□) or SalpIL2 +BC+BR oil (▪) starting on day 0 as compared to control animals (•). Notes: The table insert indicates statistical significance between groups. Data are mean ± standard deviation from one experiment, N = 5 mice per group.
Figure Legend Snippet: Effect of combination antioxidant seed oil on the splenic lymphocyte response to SalpIL2 in mice. Splenic natural killer (NK) ( A ), CD 8 + T ( B ), and CD 4 + T cell populations ( C ) as determined by flow cytometry in response in animals fed a diet consisting of equal amounts of black raspberry (BR; Rubus occidentalis ), black cumin (BC; Nigella sativa ) seed oils (○), administered a single oral dose of SalpIL2 (□) or SalpIL2 +BC+BR oil (▪) starting on day 0 as compared to control animals (•). Notes: The table insert indicates statistical significance between groups. Data are mean ± standard deviation from one experiment, N = 5 mice per group.

Techniques Used: Mouse Assay, Flow Cytometry, Cytometry, Standard Deviation

Effect of combination antioxidant seed oil on the SalpIL2 anti-tumor response in mice. On Day 0, 5 × 10 4 MCA-38 cells were injected intrasplenically to naïve animals. A single oral administration of SalpIL2 and initiation of a diet consisting of equal amounts of black raspberry (BR; Rubus occidentalis ), black cumin (BC; Nigella sativa ) seed oils 10% w/w was given to animals on Day 3. Animals were maintained for a total of 14 days prior to collection of tumor and hepatic lymphocyte data (see diagram above). Tumor number ( A ) and tumor volume ( B ) in animals fed a BC+BR with or without SalpIL2 . Hepatic natural killer (NK) ( C ), CD8 + T ( D ), and CD4 + T cell response ( E ) to SalpIL2 and antioxidant oils in tumor burden mice at the experimental endpoint. Notes: Bars indicate significance between groups. Data was normalized to percent of control and presented as ± standard deviation from one experiment with N = 4 mice per group.
Figure Legend Snippet: Effect of combination antioxidant seed oil on the SalpIL2 anti-tumor response in mice. On Day 0, 5 × 10 4 MCA-38 cells were injected intrasplenically to naïve animals. A single oral administration of SalpIL2 and initiation of a diet consisting of equal amounts of black raspberry (BR; Rubus occidentalis ), black cumin (BC; Nigella sativa ) seed oils 10% w/w was given to animals on Day 3. Animals were maintained for a total of 14 days prior to collection of tumor and hepatic lymphocyte data (see diagram above). Tumor number ( A ) and tumor volume ( B ) in animals fed a BC+BR with or without SalpIL2 . Hepatic natural killer (NK) ( C ), CD8 + T ( D ), and CD4 + T cell response ( E ) to SalpIL2 and antioxidant oils in tumor burden mice at the experimental endpoint. Notes: Bars indicate significance between groups. Data was normalized to percent of control and presented as ± standard deviation from one experiment with N = 4 mice per group.

Techniques Used: Mouse Assay, Injection, Standard Deviation

Milk thistle (MT; Silybum marianum ) seed oil inhibits SalpIL2 -dependent prevention of experimental hepatic metastases in mice. A single oral administration of SalpIL2 and initiation of MT (shaded area) seed oil diet 10% w/w was given to animals on Day –7. On Day 0, 5 × 10 4 MCA-38 cells were injected intrasplenically and animals were maintained for 14 additional days. Tumor and hepatic lymphocyte data collected on Day 21 (see diagram above). Tumor number ( A ) and tumor volume ( B ) in animals fed a diet consisting of MT oil and or SalpIL2 . Natural killer (NK) ( C ), CD8 + T ( D ), and CD4 + T cell response ( E ) to SalpIL2 and MT oil on Day 21. Notes: Bars represent statistical significance between groups. Data was normalized to percent of control and presented as ± standard deviation from one experiment with N = 5 mice per group.
Figure Legend Snippet: Milk thistle (MT; Silybum marianum ) seed oil inhibits SalpIL2 -dependent prevention of experimental hepatic metastases in mice. A single oral administration of SalpIL2 and initiation of MT (shaded area) seed oil diet 10% w/w was given to animals on Day –7. On Day 0, 5 × 10 4 MCA-38 cells were injected intrasplenically and animals were maintained for 14 additional days. Tumor and hepatic lymphocyte data collected on Day 21 (see diagram above). Tumor number ( A ) and tumor volume ( B ) in animals fed a diet consisting of MT oil and or SalpIL2 . Natural killer (NK) ( C ), CD8 + T ( D ), and CD4 + T cell response ( E ) to SalpIL2 and MT oil on Day 21. Notes: Bars represent statistical significance between groups. Data was normalized to percent of control and presented as ± standard deviation from one experiment with N = 5 mice per group.

Techniques Used: Mouse Assay, Injection, Standard Deviation

Effect of black raspberry (BR; Rubus occidentalis ) seed oil on the SalpIL2 anti-tumor response in mice. On Day 0, 5 × 10 4 MCA-38 cells were injected intrasplenically to naïve animals. A single oral administration of SalpIL2 and initiation of BR (shaded area) seed oil diet 10% w/w was given to animals on Day 3. Animals were maintained for a total of 14 days prior to collection of tumor and hepatic lymphocyte data (see diagram above). Tumor number ( A ) and tumor volume ( B ) in animals fed a diet consisting of BR with and without SalpIL2 . Hepatic natural killer (NK) ( C ), CD8 + T ( D ), and CD4 + T cell response ( E ) to SalpIL2 with and without BR oil on Day 14. Notes: Bars represent statistical significance between groups. Data was normalized to percent of control and presented as ± standard error of mean from three independent experiments with N = 5 mice per group.
Figure Legend Snippet: Effect of black raspberry (BR; Rubus occidentalis ) seed oil on the SalpIL2 anti-tumor response in mice. On Day 0, 5 × 10 4 MCA-38 cells were injected intrasplenically to naïve animals. A single oral administration of SalpIL2 and initiation of BR (shaded area) seed oil diet 10% w/w was given to animals on Day 3. Animals were maintained for a total of 14 days prior to collection of tumor and hepatic lymphocyte data (see diagram above). Tumor number ( A ) and tumor volume ( B ) in animals fed a diet consisting of BR with and without SalpIL2 . Hepatic natural killer (NK) ( C ), CD8 + T ( D ), and CD4 + T cell response ( E ) to SalpIL2 with and without BR oil on Day 14. Notes: Bars represent statistical significance between groups. Data was normalized to percent of control and presented as ± standard error of mean from three independent experiments with N = 5 mice per group.

Techniques Used: Mouse Assay, Injection

Effect of black cumin (BC; Nigella sativa ) seed oil on the SalpIL2 anti-tumor response in mice. On Day 0, 5 × 10 4 MCA-38 cells were injected intrasplenically to naïve animals. A single oral administration of SalpIL2 and initiation of BC (shaded area) seed oil diet 10% w/w was given to animals on Day 3. Animals were maintained for a total of 14 days prior to collection of tumor and hepatic lymphocyte data (see diagram above). Tumor number ( A ) and tumor volume ( B ) in animals fed a diet BC seed oil with and without SalpIL2 . Hepatic natural killer (NK) ( C ), CD8 + T ( D ), and CD4 + T cell response ( E ) to SalpIL2 with and without BC oil on Day 14. Notes: Bars represent statistical significance between groups. Data is from one experiment, normalized to percent of control, N = 5, control and SalpIL2 ; N = 10, BC oil and SalpIL2 +BC oil; and presented as ± standard deviation.
Figure Legend Snippet: Effect of black cumin (BC; Nigella sativa ) seed oil on the SalpIL2 anti-tumor response in mice. On Day 0, 5 × 10 4 MCA-38 cells were injected intrasplenically to naïve animals. A single oral administration of SalpIL2 and initiation of BC (shaded area) seed oil diet 10% w/w was given to animals on Day 3. Animals were maintained for a total of 14 days prior to collection of tumor and hepatic lymphocyte data (see diagram above). Tumor number ( A ) and tumor volume ( B ) in animals fed a diet BC seed oil with and without SalpIL2 . Hepatic natural killer (NK) ( C ), CD8 + T ( D ), and CD4 + T cell response ( E ) to SalpIL2 with and without BC oil on Day 14. Notes: Bars represent statistical significance between groups. Data is from one experiment, normalized to percent of control, N = 5, control and SalpIL2 ; N = 10, BC oil and SalpIL2 +BC oil; and presented as ± standard deviation.

Techniques Used: Mouse Assay, Injection, Standard Deviation

Effect of black raspberry (BR; Rubus occidentalis ) seed oil on the splenic lymphocyte response to SalpIL2 in mice. Splenic natural killer (NK) ( A ), CD8 + T ( B ), and CD4 + T cell populations ( C ) as determined by flow cytometry in response in animals fed a diet consisting of BR seed oil 10% w/w (○), administered as single oral dose of SalpIL2 (□) or SalpIL2 + BR oil (▪) starting on day 0 as compared to control animals (•). Notes: The table insert indicates statistical significance between groups. Data are mean ± standard deviation from one experiment, N = 5 mice group.
Figure Legend Snippet: Effect of black raspberry (BR; Rubus occidentalis ) seed oil on the splenic lymphocyte response to SalpIL2 in mice. Splenic natural killer (NK) ( A ), CD8 + T ( B ), and CD4 + T cell populations ( C ) as determined by flow cytometry in response in animals fed a diet consisting of BR seed oil 10% w/w (○), administered as single oral dose of SalpIL2 (□) or SalpIL2 + BR oil (▪) starting on day 0 as compared to control animals (•). Notes: The table insert indicates statistical significance between groups. Data are mean ± standard deviation from one experiment, N = 5 mice group.

Techniques Used: Mouse Assay, Flow Cytometry, Cytometry, Standard Deviation

Effect of black raspberry (BR; Rubus occidentalis ) seed oil and SalpIL2 on the prevention of experimental hepatic metastases in mice. A single oral administration of SalpIL2 and initiation of BR (shaded area) seed oil diet 10% w/w was given to animals on Day −7. On Day 0, 5x10 4 MCA-38 cells were injected intrasplenically and animals were maintained for 14 additional days. Tumor and hepatic lymphocyte data collected on Day 14 (see diagram above). Tumor number ( A ) and tumor volume ( B ) in animals fed a diet consisting of BR with and without SalpIL2 . Hepatic natural killer (NK) ( C ), CD8 + T ( D ), and CD4 + T cell response ( E ) to SalpIL2 and BR oil in tumor burden mice on Day 21. Notes: Bars represent statistical significance between groups. Data was normalized to percent of control and presented as ± standard error of mean from three independent experiments with N = 5 mice per group.
Figure Legend Snippet: Effect of black raspberry (BR; Rubus occidentalis ) seed oil and SalpIL2 on the prevention of experimental hepatic metastases in mice. A single oral administration of SalpIL2 and initiation of BR (shaded area) seed oil diet 10% w/w was given to animals on Day −7. On Day 0, 5x10 4 MCA-38 cells were injected intrasplenically and animals were maintained for 14 additional days. Tumor and hepatic lymphocyte data collected on Day 14 (see diagram above). Tumor number ( A ) and tumor volume ( B ) in animals fed a diet consisting of BR with and without SalpIL2 . Hepatic natural killer (NK) ( C ), CD8 + T ( D ), and CD4 + T cell response ( E ) to SalpIL2 and BR oil in tumor burden mice on Day 21. Notes: Bars represent statistical significance between groups. Data was normalized to percent of control and presented as ± standard error of mean from three independent experiments with N = 5 mice per group.

Techniques Used: Mouse Assay, Injection

31) Product Images from "Administration of defined microbiota is protective in a murine Salmonella infection model"

Article Title: Administration of defined microbiota is protective in a murine Salmonella infection model

Journal: Scientific Reports

doi: 10.1038/srep16094

MET-1 pre-treatment does not prevent S. typhimurium intracellular invasion of Caco-2 epithelial cells. Live (or heat-killed) S. typhimurium were added to Caco-2 cell monolayers at an MOI of 100:1 and incubated for 1 hour. Extracellular bacteria were then killed by the addition of gentamicin (100 μg/mL). One hour later, Caco-2 cells were lysed and intracellular bacteria were enumerated using serial dilutions plated on MacConkey agar plates containing 100 μg/mL streptomycin. There were no significant differences between S. typhimurium -infected cells pretreated with MET-1 (MET-1 + Sal) or saline (Saline + Sal) (p > 0.05). No bacterial growth was seen in cell monolayers treated with saline only (Saline), MET-1 only (MET-1), heat-killed S. typhimurium only (Saline + HK Sal) or MET-1 pretreated cell monolayers treated with heat-killed S. typhimurium (MET-1 + HK Sal). Data were analyzed using a 1-way ANOVA with Bonferroni correction, n = 3, (p > 0.05) NS = not significant.
Figure Legend Snippet: MET-1 pre-treatment does not prevent S. typhimurium intracellular invasion of Caco-2 epithelial cells. Live (or heat-killed) S. typhimurium were added to Caco-2 cell monolayers at an MOI of 100:1 and incubated for 1 hour. Extracellular bacteria were then killed by the addition of gentamicin (100 μg/mL). One hour later, Caco-2 cells were lysed and intracellular bacteria were enumerated using serial dilutions plated on MacConkey agar plates containing 100 μg/mL streptomycin. There were no significant differences between S. typhimurium -infected cells pretreated with MET-1 (MET-1 + Sal) or saline (Saline + Sal) (p > 0.05). No bacterial growth was seen in cell monolayers treated with saline only (Saline), MET-1 only (MET-1), heat-killed S. typhimurium only (Saline + HK Sal) or MET-1 pretreated cell monolayers treated with heat-killed S. typhimurium (MET-1 + HK Sal). Data were analyzed using a 1-way ANOVA with Bonferroni correction, n = 3, (p > 0.05) NS = not significant.

Techniques Used: Incubation, Infection

Translocation of S. typhimurium to the spleen was reduced in MET-1 pretreated mice. Spleens were harvested 48 hours after S. typhimurium infection, weighed, and homogenized in 1 mL PBS. Serial dilutions were plated on MacConkey agar plates containing 100 μg/mL streptomycin. Plates were incubated at 37 °C for 24 hrs, and the resulting colonies were counted. Infected mice pretreated with MET-1 (MET-1 + S . Tm., n = 13) had reduced S. typhimurium counts in the spleen (*p
Figure Legend Snippet: Translocation of S. typhimurium to the spleen was reduced in MET-1 pretreated mice. Spleens were harvested 48 hours after S. typhimurium infection, weighed, and homogenized in 1 mL PBS. Serial dilutions were plated on MacConkey agar plates containing 100 μg/mL streptomycin. Plates were incubated at 37 °C for 24 hrs, and the resulting colonies were counted. Infected mice pretreated with MET-1 (MET-1 + S . Tm., n = 13) had reduced S. typhimurium counts in the spleen (*p

Techniques Used: Translocation Assay, Mouse Assay, Infection, Incubation

32) Product Images from "Genetics of Swarming Motility in Salmonella enterica Serovar Typhimurium: Critical Role for Lipopolysaccharide"

Article Title: Genetics of Swarming Motility in Salmonella enterica Serovar Typhimurium: Critical Role for Lipopolysaccharide

Journal: Journal of Bacteriology

doi:

Rescue of swarming defects with external addition of surfactin and LPS. Five-microliter drops of solution containing surfactin or LPS were placed at the center of 0.5% Difco swarm medium, inoculated with S. enterica serovar Typhimurium ( waaL and cheA ) and E. coli (RP437) strains, and incubated overnight as described in Materials and Methods.
Figure Legend Snippet: Rescue of swarming defects with external addition of surfactin and LPS. Five-microliter drops of solution containing surfactin or LPS were placed at the center of 0.5% Difco swarm medium, inoculated with S. enterica serovar Typhimurium ( waaL and cheA ) and E. coli (RP437) strains, and incubated overnight as described in Materials and Methods.

Techniques Used: Incubation

33) Product Images from "Commensal Akkermansia muciniphila Exacerbates Gut Inflammation in Salmonella Typhimurium-Infected Gnotobiotic Mice"

Article Title: Commensal Akkermansia muciniphila Exacerbates Gut Inflammation in Salmonella Typhimurium-Infected Gnotobiotic Mice

Journal: PLoS ONE

doi: 10.1371/journal.pone.0074963

SIHUMI mice colonized with both A. muciniphila and S. Typhimurium display an increased cecal macrophage infiltration. (A) Formalin fixed paraffin embedded cecum tissue was thin sectioned at 2 µm. Macrophages were stained by targeting the F4/80 receptor expressed on mouse macrophages using immunohistochemistry with specific antibodies. Brown color indicates positively stained macrophages. Magnification 400-fold. Bar = 100 µm. (B) Positively stained macrophages were enumerated along a stretch of 50 µm of lamina muscularis for both lamina propria and sub-mucosa (see materials and methods). SIHUMI mice colonized with both A. muciniphila and S. Typhimurium had the highest macrophage infiltration scores compared to the other groups (see Figure. 1). Data are expressed as median with range. n = 5 mice per group. *P
Figure Legend Snippet: SIHUMI mice colonized with both A. muciniphila and S. Typhimurium display an increased cecal macrophage infiltration. (A) Formalin fixed paraffin embedded cecum tissue was thin sectioned at 2 µm. Macrophages were stained by targeting the F4/80 receptor expressed on mouse macrophages using immunohistochemistry with specific antibodies. Brown color indicates positively stained macrophages. Magnification 400-fold. Bar = 100 µm. (B) Positively stained macrophages were enumerated along a stretch of 50 µm of lamina muscularis for both lamina propria and sub-mucosa (see materials and methods). SIHUMI mice colonized with both A. muciniphila and S. Typhimurium had the highest macrophage infiltration scores compared to the other groups (see Figure. 1). Data are expressed as median with range. n = 5 mice per group. *P

Techniques Used: Mouse Assay, Formalin-fixed Paraffin-Embedded, Staining, Immunohistochemistry

Concomitant presence of A. muciniphila and S. Typhimurium results in increased histopathology scores in SIHUMI mice. (A) Gnotobiotic C3H mice containing 8 defined microbial species (SIHUMI) were subsequently inoculated with A. muciniphila or S. Typhimurium or consecutively with both organisms (see Figure 1 ). SIHUMI and SIHUMI-A mice had the lowest histopathology scores (≤4.0) with no signs of inflammation and were therefore taken as baseline (dotted line). Data are expressed as median with range. *P
Figure Legend Snippet: Concomitant presence of A. muciniphila and S. Typhimurium results in increased histopathology scores in SIHUMI mice. (A) Gnotobiotic C3H mice containing 8 defined microbial species (SIHUMI) were subsequently inoculated with A. muciniphila or S. Typhimurium or consecutively with both organisms (see Figure 1 ). SIHUMI and SIHUMI-A mice had the lowest histopathology scores (≤4.0) with no signs of inflammation and were therefore taken as baseline (dotted line). Data are expressed as median with range. *P

Techniques Used: Histopathology, Mouse Assay

SIHUMI mice colonized with both A. muciniphila and S. Typhimurium display enlarged mLN and elevated S. Typhimurium cell numbers. (A) Mesenteric lymph nodes (mLN) were obtained from four groups of gnotobiotic C3H mice. SIHUMI mice were subsequently inoculated with A. muciniphila or S. Typhimurium or consecutively with both organisms (see Figure 1 ). The mLN tissue was homogenized and DNA was isolated to quantify S. Typhimurium using quantitative PCR with primers targeting the ttr-region of S. Typhimurium. Absolute cell numbers were calculated based on calibration curves with known concentrations of S. Typhimurium. The mLN of SIHUMI-AS mice contained 10-fold higher cell numbers of S. Typhimurium compared to SIHUMI-S mice. Data are expressed as mean±standard error. n = 10 mice per group. *P
Figure Legend Snippet: SIHUMI mice colonized with both A. muciniphila and S. Typhimurium display enlarged mLN and elevated S. Typhimurium cell numbers. (A) Mesenteric lymph nodes (mLN) were obtained from four groups of gnotobiotic C3H mice. SIHUMI mice were subsequently inoculated with A. muciniphila or S. Typhimurium or consecutively with both organisms (see Figure 1 ). The mLN tissue was homogenized and DNA was isolated to quantify S. Typhimurium using quantitative PCR with primers targeting the ttr-region of S. Typhimurium. Absolute cell numbers were calculated based on calibration curves with known concentrations of S. Typhimurium. The mLN of SIHUMI-AS mice contained 10-fold higher cell numbers of S. Typhimurium compared to SIHUMI-S mice. Data are expressed as mean±standard error. n = 10 mice per group. *P

Techniques Used: Mouse Assay, Isolation, Real-time Polymerase Chain Reaction

SIHUMI mice colonized with both A. muciniphila and S. Typhimurium display reduced mucus sulphation. Formalin fixed thin sections (4 µm) of cecal tissue of mice belonging to either one of four groups: SIHUMI, SIHUMI-A, SIHUMI-S and SIHUMI-AS (see Figure. 1) were stained with high iron diamine (HID)/AB at pH-2.5 and subsequently analyzed. Brown color indicates sulphated mucins while blue color indicates sialylated mucins. SIHUMI-AS mice display few sulphated mucins compared to the other mouse groups. Magnification 400×. Bars indicate 100 µm.
Figure Legend Snippet: SIHUMI mice colonized with both A. muciniphila and S. Typhimurium display reduced mucus sulphation. Formalin fixed thin sections (4 µm) of cecal tissue of mice belonging to either one of four groups: SIHUMI, SIHUMI-A, SIHUMI-S and SIHUMI-AS (see Figure. 1) were stained with high iron diamine (HID)/AB at pH-2.5 and subsequently analyzed. Brown color indicates sulphated mucins while blue color indicates sialylated mucins. SIHUMI-AS mice display few sulphated mucins compared to the other mouse groups. Magnification 400×. Bars indicate 100 µm.

Techniques Used: Mouse Assay, Staining

Hypothetical Scheme. The presence of A. muciniphila, leads to the exacerbation of S. Typhimurium-induced intestinal inflammation. We propose that the presence of A. muciniphila causes changes in mucin composition and production, which in turn facilitates the invasion of S. Typhimurium into the host. Increased inflammatory status was characterized by increased pro-inflammatory cytokines, increased macrophage infiltration and invasion of the pathogen into the lymph nodes, reduced number of mucin-filled goblet cells in SIHUMI-AS mice (B) compared to SIHUMI-S mice (A). Our data suggests that in the presence of both A. muciniphila and S. Typhimurium, mucus sulphation is diminished and this may facilitate the access of S. Typhimurium to sialic acid in mucus. Sialic acid may serve as a substrate and adhesion site for S. Typhimurium in the gut [56] , [57] . Increased gene expression of IFN-γ and IP-10 indicate an increased NK-cell recruitment. mLN - mesenteric lymph nodes, NK- Natural killer cells. (↑ increased; ↓ decreased; grey dotted line: assumed processes including lectin-sialic acid binding [56] , M-cells for pathogen transit [43] , [58] , [59] ; black line: supported by data of the present study).
Figure Legend Snippet: Hypothetical Scheme. The presence of A. muciniphila, leads to the exacerbation of S. Typhimurium-induced intestinal inflammation. We propose that the presence of A. muciniphila causes changes in mucin composition and production, which in turn facilitates the invasion of S. Typhimurium into the host. Increased inflammatory status was characterized by increased pro-inflammatory cytokines, increased macrophage infiltration and invasion of the pathogen into the lymph nodes, reduced number of mucin-filled goblet cells in SIHUMI-AS mice (B) compared to SIHUMI-S mice (A). Our data suggests that in the presence of both A. muciniphila and S. Typhimurium, mucus sulphation is diminished and this may facilitate the access of S. Typhimurium to sialic acid in mucus. Sialic acid may serve as a substrate and adhesion site for S. Typhimurium in the gut [56] , [57] . Increased gene expression of IFN-γ and IP-10 indicate an increased NK-cell recruitment. mLN - mesenteric lymph nodes, NK- Natural killer cells. (↑ increased; ↓ decreased; grey dotted line: assumed processes including lectin-sialic acid binding [56] , M-cells for pathogen transit [43] , [58] , [59] ; black line: supported by data of the present study).

Techniques Used: Mouse Assay, Expressing, Binding Assay

Presence of A. muciniphila renders S. Typhimurium the dominant species in gnotobiotic SIHUMI mice. Cecal contents were collected from gnotobiotic C3H mice, differing in their microbial status: (A) Mice with a defined microbial community of eight bacterial species (SIHUMI), (B) SIHUMI mice additionally colonized with A. muciniphila (SIHUMI-A), (C) SIHUMI mice infected with S. Typhimurium (SIHUMI-S) and (D) SIHUMI mice colonized with A. muciniphila and 10 days later infected with S. Typhimurium (SIHUMI-AS) (see Figure 1 ). Total DNA was extracted and bacterial cell numbers were quantified by qPCR with primers targeting the HSP60 gene of the SIHUMI members, the 16S rRNA gene of A. muciniphila and the ttr-region of S. Typhimurium. Calculation of the cell numbers was based on DNA obtained from cell suspensions containing known cell numbers of the targeted bacterial species (see materials and methods). Presence of A. muciniphila in SIHUMI-AS mice is attributed to an increase in the proportion of S. Typhimurium cells at the expense of other community members showing reduced proportion of SIHUMI members. Ten animals per group were used. The bacterial cell numbers and P-values for the differences between the groups are provided in Table 1 .
Figure Legend Snippet: Presence of A. muciniphila renders S. Typhimurium the dominant species in gnotobiotic SIHUMI mice. Cecal contents were collected from gnotobiotic C3H mice, differing in their microbial status: (A) Mice with a defined microbial community of eight bacterial species (SIHUMI), (B) SIHUMI mice additionally colonized with A. muciniphila (SIHUMI-A), (C) SIHUMI mice infected with S. Typhimurium (SIHUMI-S) and (D) SIHUMI mice colonized with A. muciniphila and 10 days later infected with S. Typhimurium (SIHUMI-AS) (see Figure 1 ). Total DNA was extracted and bacterial cell numbers were quantified by qPCR with primers targeting the HSP60 gene of the SIHUMI members, the 16S rRNA gene of A. muciniphila and the ttr-region of S. Typhimurium. Calculation of the cell numbers was based on DNA obtained from cell suspensions containing known cell numbers of the targeted bacterial species (see materials and methods). Presence of A. muciniphila in SIHUMI-AS mice is attributed to an increase in the proportion of S. Typhimurium cells at the expense of other community members showing reduced proportion of SIHUMI members. Ten animals per group were used. The bacterial cell numbers and P-values for the differences between the groups are provided in Table 1 .

Techniques Used: Mouse Assay, Infection, Real-time Polymerase Chain Reaction

Presence of both A. muciniphila and S. Typhimurium is accompanied by increased pro-inflammatory cytokines. (A) Cecal mRNA levels of IFN-γ, IP-10, TNF-α, IL-12, IL-6, IL-17 and IL-18 in gnotobiotic SIHUMI mice were measured. mRNA was extracted from cecum mucosa of mice belonging to either one of four groups: SIHUMI, SIHUMI-A, SIHUMI-S and SIHUMI-AS (see Figure. 1). The mRNA was converted to cDNA for quantitative real-time PCR measurement (see materials and methods). Inoculation of the gnotobiotic SIHUMI mice with A. muciniphila followed by S. Typhimurium infection (SIHUMI-AS) caused an increase in mRNA levels of pro-inflammatory cytokines except IL-18. Data are expressed as mean±standard error. n = 6 per group. Star indicates statistically significant differences ( *P
Figure Legend Snippet: Presence of both A. muciniphila and S. Typhimurium is accompanied by increased pro-inflammatory cytokines. (A) Cecal mRNA levels of IFN-γ, IP-10, TNF-α, IL-12, IL-6, IL-17 and IL-18 in gnotobiotic SIHUMI mice were measured. mRNA was extracted from cecum mucosa of mice belonging to either one of four groups: SIHUMI, SIHUMI-A, SIHUMI-S and SIHUMI-AS (see Figure. 1). The mRNA was converted to cDNA for quantitative real-time PCR measurement (see materials and methods). Inoculation of the gnotobiotic SIHUMI mice with A. muciniphila followed by S. Typhimurium infection (SIHUMI-AS) caused an increase in mRNA levels of pro-inflammatory cytokines except IL-18. Data are expressed as mean±standard error. n = 6 per group. Star indicates statistically significant differences ( *P

Techniques Used: Mouse Assay, Real-time Polymerase Chain Reaction, Infection

SIHUMI mice with both A. muciniphila and S. Typhimurium display increased MUC2 mRNA levels (A) and reduced numbers of mucin filled goblet cells (B and C). (A) mRNA was extracted from cecum mucosa of mice belonging to either one of four groups: SIHUMI, SIHUMI-A, SIHUMI-S and SIHUMI-AS. MUC2 mRNA from cecum mucosa was converted to cDNA and expression levels were quantified using real-time PCR (see materials and methods). SIHUMI-A and SIHUMI-AS mice showed significantly higher MUC2 gene expression compared to the other two groups, harboring no A. muciniphila . Data are expressed as mean±standard error. n = 6 per group. *P
Figure Legend Snippet: SIHUMI mice with both A. muciniphila and S. Typhimurium display increased MUC2 mRNA levels (A) and reduced numbers of mucin filled goblet cells (B and C). (A) mRNA was extracted from cecum mucosa of mice belonging to either one of four groups: SIHUMI, SIHUMI-A, SIHUMI-S and SIHUMI-AS. MUC2 mRNA from cecum mucosa was converted to cDNA and expression levels were quantified using real-time PCR (see materials and methods). SIHUMI-A and SIHUMI-AS mice showed significantly higher MUC2 gene expression compared to the other two groups, harboring no A. muciniphila . Data are expressed as mean±standard error. n = 6 per group. *P

Techniques Used: Mouse Assay, Expressing, Real-time Polymerase Chain Reaction

Design of the animal experiment. Fourty C3H mice associated with a defined microbial community of 8 bacterial species (SIHUMI) were allocated to four different groups (10 mice per group). Each mouse was associated with 8 bacterial species (SIHUMI). Twelve weeks-old SIHUMI mice were subsequently associated with A. muciniphila (SIHUMI-A) or S. Typhimurium (SIHUMI-S) or with both A. muciniphila and S. Typhimurium (SIHUMI-AS). SIHUMI mice received only sterile medium. Times of association, infection and killing are as indicated. ‡ - killed.
Figure Legend Snippet: Design of the animal experiment. Fourty C3H mice associated with a defined microbial community of 8 bacterial species (SIHUMI) were allocated to four different groups (10 mice per group). Each mouse was associated with 8 bacterial species (SIHUMI). Twelve weeks-old SIHUMI mice were subsequently associated with A. muciniphila (SIHUMI-A) or S. Typhimurium (SIHUMI-S) or with both A. muciniphila and S. Typhimurium (SIHUMI-AS). SIHUMI mice received only sterile medium. Times of association, infection and killing are as indicated. ‡ - killed.

Techniques Used: Mouse Assay, Infection

34) Product Images from "In vitro and in vivo probiotic assessment of Leuconostoc mesenteroides P45 isolated from pulque, a Mexican traditional alcoholic beverage"

Article Title: In vitro and in vivo probiotic assessment of Leuconostoc mesenteroides P45 isolated from pulque, a Mexican traditional alcoholic beverage

Journal: SpringerPlus

doi: 10.1186/s40064-016-2370-7

Anti-infective effect of L. mesenteroides P45 on Salmonella enterica serovar Typhimurium str r in mouse liver and spleen. LC liver control, LE liver experimental, SC spleen control, SE spleen experimental. Values are the mean log 10 CFU/mL count from liver and spleen samples obtained from nine mice per group
Figure Legend Snippet: Anti-infective effect of L. mesenteroides P45 on Salmonella enterica serovar Typhimurium str r in mouse liver and spleen. LC liver control, LE liver experimental, SC spleen control, SE spleen experimental. Values are the mean log 10 CFU/mL count from liver and spleen samples obtained from nine mice per group

Techniques Used: Mouse Assay

Antimicrobial activity of L. mesenteroides P45 against enteropathogenic Escherichia coli , Salmonella enterica serovar Typhimurium, S. enterica serovar Typhimurium and L. monocytogenes . Upper panel cell-to-cell antimicrobial effect of a lawn of strain P45. Middle panel 100 µL of cell-free, neutralized and 2× concentrated supernatant. Bottom panel cell-to-cell antimicrobial effect of a lawn of EPS-producing P45 grown on APT + 20 % sucrose. Mean ± SD inhibition zones (mm) observed for upper panel were: 11.75 ± 1.26, 11.25 ± 1.73, 9.25 ± 0.82, 10.5 ± 1.63, for each assayed bacteria, respectively. For middle panel : 7.5, 6.7 ± 0.5, 6.5 ± 0.58, 9.0 ± 1.15, respectively. Values of inhibition zone in EPS-producing cell-to-cell assays are omitted because the heterogeneous shape of the lawn of strain P45 ( bottom panel )
Figure Legend Snippet: Antimicrobial activity of L. mesenteroides P45 against enteropathogenic Escherichia coli , Salmonella enterica serovar Typhimurium, S. enterica serovar Typhimurium and L. monocytogenes . Upper panel cell-to-cell antimicrobial effect of a lawn of strain P45. Middle panel 100 µL of cell-free, neutralized and 2× concentrated supernatant. Bottom panel cell-to-cell antimicrobial effect of a lawn of EPS-producing P45 grown on APT + 20 % sucrose. Mean ± SD inhibition zones (mm) observed for upper panel were: 11.75 ± 1.26, 11.25 ± 1.73, 9.25 ± 0.82, 10.5 ± 1.63, for each assayed bacteria, respectively. For middle panel : 7.5, 6.7 ± 0.5, 6.5 ± 0.58, 9.0 ± 1.15, respectively. Values of inhibition zone in EPS-producing cell-to-cell assays are omitted because the heterogeneous shape of the lawn of strain P45 ( bottom panel )

Techniques Used: Activity Assay, Inhibition

35) Product Images from "Benzyl Isothiocyanate, a Major Component from the Roots of Salvadora Persica Is Highly Active against Gram-Negative Bacteria"

Article Title: Benzyl Isothiocyanate, a Major Component from the Roots of Salvadora Persica Is Highly Active against Gram-Negative Bacteria

Journal: PLoS ONE

doi: 10.1371/journal.pone.0023045

Antibacterial activity of S. persica essential oil determined by colony forming units (CFU) assay. The individual bars show the statistical mean and standard deviation of the number of surviving bacteria from three experiments. The essential oil concentration is in percentage of final assay volume. Control is with DMSO only. (A) Gram-negative bacteria (B) Gram-positive bacteria (C) Kinetics of antibacterial activity against E. coli MC4100 (Ec) and A. actinomycetemcomitans (Aa) Samples were withdrawn for CFU determination at time-points indicated. Oil dilution was 0.1%. The CFU values displayed are the statistical mean of three experiments.
Figure Legend Snippet: Antibacterial activity of S. persica essential oil determined by colony forming units (CFU) assay. The individual bars show the statistical mean and standard deviation of the number of surviving bacteria from three experiments. The essential oil concentration is in percentage of final assay volume. Control is with DMSO only. (A) Gram-negative bacteria (B) Gram-positive bacteria (C) Kinetics of antibacterial activity against E. coli MC4100 (Ec) and A. actinomycetemcomitans (Aa) Samples were withdrawn for CFU determination at time-points indicated. Oil dilution was 0.1%. The CFU values displayed are the statistical mean of three experiments.

Techniques Used: Activity Assay, Colony-forming Unit Assay, Standard Deviation, Concentration Assay

36) Product Images from "Ca2+-dependent Focal Exocytosis of Golgi-derived Vesicles Helps Phagocytic Uptake in Macrophages *"

Article Title: Ca2+-dependent Focal Exocytosis of Golgi-derived Vesicles Helps Phagocytic Uptake in Macrophages *

Journal: The Journal of Biological Chemistry

doi: 10.1074/jbc.M116.743047

Mannosidase-II is recruited at the site of phagocytosis. A, RAW264.7 macrophages expressing mCherry-MAN2A ( red ) were incubated with mouse serum-coated latex beads for 30 min and 1 h. The images shown are representative of the 30-min samples. Right , total intensity of mCherry-MAN2A puncta on the bead surface was estimated using 3D spot creation module in Imaris 7.2, and the intensity distribution of the population has been plotted (see “Materials and Methods” for detail). The box plot represents data from more than 200 beads analyzed from two different experiments (values ± S.D.; scale bar, 4 μm). B, THP-1-derived macrophages were infected with GFP expressing E. coli ( green, top panel ) or S. typhimurium ( green, lower panel ) for 15 and 30 min. Cells were then stained with anti-mannosidase-II antibody followed by Alexa Fluor 568-tagged secondary antibody ( red ). Images shown are representative of the 15-min samples from both experiments. The bar plots at right show % co-localization of E. coli or Salmonella with mannosidase-II at the surface at 5 and 10 or 15 and 30 min, respectively, post-infection. Data represent the average of more than 200 bacteria from two different experiments (values ± S.D.; scale bar, 3 μm). C, mCherry-MAN2A ( red )-expressing THP-1-derived macrophages were infected with PKH67-labeled H37Rv ( green ) for 1 and 2 h. Samples were fixed and visualized under the microscope. The bar plot at right shows % co-localization of H37Rv with mannosidase-II at both these time points. Data represent average of more than 200 bacteria from two different experiments (values ± S.D.; scale bar, 4 μm). D, THP-1-derived macrophages were infected with PKH67-labeled H37RV and fixed at the respective time points. These cells were then permeabilized using 0.05% Triton X-100 and stained with anti-mannosidase-II antibody followed by the secondary antibody tagged with Alexa Fluor 568, fixed, and visualized under the confocal microscope ( scale bar, 3 μm). E, mouse BMDMs were infected with GFP expressing E. coli for 10 min, fixed, and stained with anti-MAN2A antibody ( scale bar, 3 μm). DIC , differential interference contrast.
Figure Legend Snippet: Mannosidase-II is recruited at the site of phagocytosis. A, RAW264.7 macrophages expressing mCherry-MAN2A ( red ) were incubated with mouse serum-coated latex beads for 30 min and 1 h. The images shown are representative of the 30-min samples. Right , total intensity of mCherry-MAN2A puncta on the bead surface was estimated using 3D spot creation module in Imaris 7.2, and the intensity distribution of the population has been plotted (see “Materials and Methods” for detail). The box plot represents data from more than 200 beads analyzed from two different experiments (values ± S.D.; scale bar, 4 μm). B, THP-1-derived macrophages were infected with GFP expressing E. coli ( green, top panel ) or S. typhimurium ( green, lower panel ) for 15 and 30 min. Cells were then stained with anti-mannosidase-II antibody followed by Alexa Fluor 568-tagged secondary antibody ( red ). Images shown are representative of the 15-min samples from both experiments. The bar plots at right show % co-localization of E. coli or Salmonella with mannosidase-II at the surface at 5 and 10 or 15 and 30 min, respectively, post-infection. Data represent the average of more than 200 bacteria from two different experiments (values ± S.D.; scale bar, 3 μm). C, mCherry-MAN2A ( red )-expressing THP-1-derived macrophages were infected with PKH67-labeled H37Rv ( green ) for 1 and 2 h. Samples were fixed and visualized under the microscope. The bar plot at right shows % co-localization of H37Rv with mannosidase-II at both these time points. Data represent average of more than 200 bacteria from two different experiments (values ± S.D.; scale bar, 4 μm). D, THP-1-derived macrophages were infected with PKH67-labeled H37RV and fixed at the respective time points. These cells were then permeabilized using 0.05% Triton X-100 and stained with anti-mannosidase-II antibody followed by the secondary antibody tagged with Alexa Fluor 568, fixed, and visualized under the confocal microscope ( scale bar, 3 μm). E, mouse BMDMs were infected with GFP expressing E. coli for 10 min, fixed, and stained with anti-MAN2A antibody ( scale bar, 3 μm). DIC , differential interference contrast.

Techniques Used: Expressing, Incubation, Derivative Assay, Infection, Staining, Labeling, Microscopy

37) Product Images from "Regulated Delayed Expression of rfaH in an Attenuated Salmonellaenterica Serovar Typhimurium Vaccine Enhances Immunogenicity of Outer Membrane Proteins and a Heterologous Antigen ▿"

Article Title: Regulated Delayed Expression of rfaH in an Attenuated Salmonellaenterica Serovar Typhimurium Vaccine Enhances Immunogenicity of Outer Membrane Proteins and a Heterologous Antigen ▿

Journal: Infection and Immunity

doi: 10.1128/IAI.00831-09

PspA and LacI synthesis is regulated by 0.1% arabinose. The Western blots show the synthesis of PspA in S. Typhimurium strains χ9884(pYA4088) (Δ rfaH49 ), χ9852(pYA4088) (ΔP rfaH178 ), χ9421(pYA4088) (RfaH +
Figure Legend Snippet: PspA and LacI synthesis is regulated by 0.1% arabinose. The Western blots show the synthesis of PspA in S. Typhimurium strains χ9884(pYA4088) (Δ rfaH49 ), χ9852(pYA4088) (ΔP rfaH178 ), χ9421(pYA4088) (RfaH +

Techniques Used: Western Blot

Arabinose regulation of rfaH . (A) Map of deletion-insertion mutations resulting in arabinose-regulated rfaH expression. (B) LPS phenotypes of wild-type S. Typhimurium χ3761 and the indicated isogenic derivatives. LPSs from different mutant strains
Figure Legend Snippet: Arabinose regulation of rfaH . (A) Map of deletion-insertion mutations resulting in arabinose-regulated rfaH expression. (B) LPS phenotypes of wild-type S. Typhimurium χ3761 and the indicated isogenic derivatives. LPSs from different mutant strains

Techniques Used: Expressing, Mutagenesis

Colonization of mouse spleens and livers by attenuated S. Typhimurium harboring plasmid pYA4088 grown in LB broth containing 0.1% arabinose. Shown is spleen (A) and liver (B) colonization by the indicated strains in BALB/c mice at 4 and 8 days
Figure Legend Snippet: Colonization of mouse spleens and livers by attenuated S. Typhimurium harboring plasmid pYA4088 grown in LB broth containing 0.1% arabinose. Shown is spleen (A) and liver (B) colonization by the indicated strains in BALB/c mice at 4 and 8 days

Techniques Used: Plasmid Preparation, Mouse Assay

Serum IgG responses in immunized and control mice. Total serum IgGs specific for rPspA (A), S. Typhimurium LPS (B), and SOMP (C) were measured by ELISA. The data represent reciprocal anti-IgG antibody levels in pooled sera from mice orally immunized with
Figure Legend Snippet: Serum IgG responses in immunized and control mice. Total serum IgGs specific for rPspA (A), S. Typhimurium LPS (B), and SOMP (C) were measured by ELISA. The data represent reciprocal anti-IgG antibody levels in pooled sera from mice orally immunized with

Techniques Used: Mouse Assay, Enzyme-linked Immunosorbent Assay

38) Product Images from "Repulsion and Metabolic Switches in the Collective Behavior of Bacterial Colonies"

Article Title: Repulsion and Metabolic Switches in the Collective Behavior of Bacterial Colonies

Journal: Biophysical Journal

doi: 10.1016/j.bpj.2009.04.018

The contours of arrest experimentally observed for two and four colonies of S. typhimurium and the theoretical prediction superimposed as full lines.
Figure Legend Snippet: The contours of arrest experimentally observed for two and four colonies of S. typhimurium and the theoretical prediction superimposed as full lines.

Techniques Used:

Dependency of the lag time on the initial bacterial density and nutrient concentration. ( a ) Distance traveled by the bacterial front in S. typhimurium colonies. ( Inset ) Data have been collapsed onto a single curve by a horizontal shift of −1 h
Figure Legend Snippet: Dependency of the lag time on the initial bacterial density and nutrient concentration. ( a ) Distance traveled by the bacterial front in S. typhimurium colonies. ( Inset ) Data have been collapsed onto a single curve by a horizontal shift of −1 h

Techniques Used: Concentration Assay

Repulsion versus merging of bacterial colonies. ( Upper row ) S. typhimurium at 30°C in medium A. ( a ) Repulsion with 0.4% glucose and 0.6% Eiken agar, after 24 h. ( b ) Merging with 0.05% glucose and 0.4% Eiken agar after 24 h. ( c ) Repulsion in P.
Figure Legend Snippet: Repulsion versus merging of bacterial colonies. ( Upper row ) S. typhimurium at 30°C in medium A. ( a ) Repulsion with 0.4% glucose and 0.6% Eiken agar, after 24 h. ( b ) Merging with 0.05% glucose and 0.4% Eiken agar after 24 h. ( c ) Repulsion in P.

Techniques Used:

39) Product Images from "Effect of atmospheric pressure plasma jet on the foodborne pathogens attached to commercial food containers"

Article Title: Effect of atmospheric pressure plasma jet on the foodborne pathogens attached to commercial food containers

Journal: Journal of Food Science and Technology

doi: 10.1007/s13197-015-2003-0

Salmonella Typhimurium counts (log CFU/cm 2 ) in the biofilms treated with atmospheric pressure plasma jet for 0, 5, and 10 min. a Collagen casing, b Polyethylene terephthalate, c Polypropylene. ( a-c Values with different letters within the same
Figure Legend Snippet: Salmonella Typhimurium counts (log CFU/cm 2 ) in the biofilms treated with atmospheric pressure plasma jet for 0, 5, and 10 min. a Collagen casing, b Polyethylene terephthalate, c Polypropylene. ( a-c Values with different letters within the same

Techniques Used:

40) Product Images from "Genetics of Swarming Motility in Salmonella enterica Serovar Typhimurium: Critical Role for Lipopolysaccharide"

Article Title: Genetics of Swarming Motility in Salmonella enterica Serovar Typhimurium: Critical Role for Lipopolysaccharide

Journal: Journal of Bacteriology

doi:

Flagellin levels in a Mot − mutant in broth and in swarm medium. Cultures of the indicated strains were grown either in LB broth or on Difco swarm medium as described in Materials and Methods. Aliquots of OD 600 -equalized cells were analyzed by Western blotting using antiflagellin antibodies as described in Materials and Methods.
Figure Legend Snippet: Flagellin levels in a Mot − mutant in broth and in swarm medium. Cultures of the indicated strains were grown either in LB broth or on Difco swarm medium as described in Materials and Methods. Aliquots of OD 600 -equalized cells were analyzed by Western blotting using antiflagellin antibodies as described in Materials and Methods.

Techniques Used: Mutagenesis, Western Blot

Related Articles

other:

Article Title: ?-Lactam Effects on Mixed Cultures of Common Respiratory Isolates as an Approach to Treatment Effects on Nasopharyngeal Bacterial Population Dynamics
Article Snippet: Preparation of individual and mixed cultures The strains were grown overnight on Mueller-Hinton agar (Difco laboratories) supplemented with 5% lysed sheep blood (Biomedics) (MHA) in the case of S. pneumoniae and S. pyogenes , or on GC agar (Difco laboratories) supplemented with 5% sheep blood (added at 50°C) and VX growth factors (GCSA) in the case of H. influenzae .

Article Title: Multifunctional Role of 35 Kilodalton Hyaluronan in Promoting Defense of the Intestinal Epithelium
Article Snippet: Serial dilutions were plated onto MacConkey agar supplemented with streptomycin to reduce the contribution of the confounding commensal bacteria.

Cell Culture:

Article Title: Commensal Akkermansia muciniphila Exacerbates Gut Inflammation in Salmonella Typhimurium-Infected Gnotobiotic Mice
Article Snippet: .. The SIHUMI members and S. Typhimurium were cultured in yeast casitone fatty acid (YCFA) medium while A. muciniphila was cultured in Columbia broth (Difco). .. All strains were cultured under strictly anoxic conditions using N2: CO2 (80∶20; v∶v) as the gas phase.

Article Title: IL-22 Defect During Streptococcus pneumoniae Infection Triggers Exacerbation of Chronic Obstructive Pulmonary Disease
Article Snippet: .. Cells (3 × 106 in 1 ml) were cultured in RPMI1640 (GIBCO, Invitrogen Corporation) supplemented with 10% FCS, 200 U/ml penicillin/streptomycin (PS) and then exposed to S. pneumoniae (Sp, MOI = 2) or to phytohemagglutinin (1 μg/ml) (PHA, Difco) as a positive control. ..

Positive Control:

Article Title: IL-22 Defect During Streptococcus pneumoniae Infection Triggers Exacerbation of Chronic Obstructive Pulmonary Disease
Article Snippet: .. Cells (3 × 106 in 1 ml) were cultured in RPMI1640 (GIBCO, Invitrogen Corporation) supplemented with 10% FCS, 200 U/ml penicillin/streptomycin (PS) and then exposed to S. pneumoniae (Sp, MOI = 2) or to phytohemagglutinin (1 μg/ml) (PHA, Difco) as a positive control. ..

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  • 88
    Difco s enterica serotype typhimurium
    Intracellular proliferation of S. <t>enterica</t> serotype <t>Typhimurium,</t> S. bongori , and S. bongori (pB6). (A) RAW 264.7 macrophages were infected with S. enterica serotype Typhimurium wild type, S. bongori , or S. bongori harboring pB6 encoding the SPI2 locus. All strains harbored p2812 for SPI2-dependent expression of sifA ::M45 and constitutive expression of GFP. Host cells were fixed 16 h after infection, and F-actin was stained by Texas red-phalloidin. Representative micrographs demonstrating the extent of intracellular proliferation are shown. (B) Macrophages were infected at a multiplicity of infection of ∼1 (for S. enterica serotype Typhimurium wild type [S. T. WT]) and 50 (for S. bongori [S. B.] strains), respectively. At 2 and 16 h after infection, macrophages were lysed by the addition of 0.1% Triton X-100, and serial dilutions were plated onto Mueller-Hinton agar for the enumeration of intracellular bacteria. The intracellular CFU was determined for both time points, and the rates of intracellular replication were expressed as the fold increase in CFU. Experiments were performed in triplicate on three different occasions.
    S Enterica Serotype Typhimurium, supplied by Difco, used in various techniques. Bioz Stars score: 88/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Difco s typhimurium strains
    Gene expression in S. <t>Typhimurium</t> and C. albicans . (A) Transcription of sopB and sipB of the S. Typhimurium wild type in the presence of supernatants from filaments or yeast-form C. albicans . (B) Transcriptional levels of TEC1 , HWP1 , ALS3 , and CDC42 in
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    Difco s enterica serovar typhimurium sh5014
    Effect of S. <t>enterica</t> <t>serovar</t> <t>Typhimurium</t> ( S.ty ) porins on Raf-1 (A) and MEK1/2 (B) activation after different stimulation times. Proteins from cell lysates (3 × 10 6 cells/ml) were analyzed by SDS-15% PAGE and immunoblotted with a Raf-1- or MEK1/2-phosphospecific antibody. Shifts in band mobility on SDS-PAGE due to phosphorylation were obtained with anti-Raf-1, anti-MEK1, and anti-MEK2. Fold increases in Raf-1 and MEK1/2 activation, compared to unstimulated cells (control), are shown below each lane for each blot. BSA represents a control of an unspecific stimulus.
    S Enterica Serovar Typhimurium Sh5014, supplied by Difco, used in various techniques. Bioz Stars score: 85/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Intracellular proliferation of S. enterica serotype Typhimurium, S. bongori , and S. bongori (pB6). (A) RAW 264.7 macrophages were infected with S. enterica serotype Typhimurium wild type, S. bongori , or S. bongori harboring pB6 encoding the SPI2 locus. All strains harbored p2812 for SPI2-dependent expression of sifA ::M45 and constitutive expression of GFP. Host cells were fixed 16 h after infection, and F-actin was stained by Texas red-phalloidin. Representative micrographs demonstrating the extent of intracellular proliferation are shown. (B) Macrophages were infected at a multiplicity of infection of ∼1 (for S. enterica serotype Typhimurium wild type [S. T. WT]) and 50 (for S. bongori [S. B.] strains), respectively. At 2 and 16 h after infection, macrophages were lysed by the addition of 0.1% Triton X-100, and serial dilutions were plated onto Mueller-Hinton agar for the enumeration of intracellular bacteria. The intracellular CFU was determined for both time points, and the rates of intracellular replication were expressed as the fold increase in CFU. Experiments were performed in triplicate on three different occasions.

    Journal: Infection and Immunity

    Article Title: Functional Transfer of Salmonella Pathogenicity Island 2 to Salmonella bongori and Escherichia coli

    doi: 10.1128/IAI.72.5.2879-2888.2004

    Figure Lengend Snippet: Intracellular proliferation of S. enterica serotype Typhimurium, S. bongori , and S. bongori (pB6). (A) RAW 264.7 macrophages were infected with S. enterica serotype Typhimurium wild type, S. bongori , or S. bongori harboring pB6 encoding the SPI2 locus. All strains harbored p2812 for SPI2-dependent expression of sifA ::M45 and constitutive expression of GFP. Host cells were fixed 16 h after infection, and F-actin was stained by Texas red-phalloidin. Representative micrographs demonstrating the extent of intracellular proliferation are shown. (B) Macrophages were infected at a multiplicity of infection of ∼1 (for S. enterica serotype Typhimurium wild type [S. T. WT]) and 50 (for S. bongori [S. B.] strains), respectively. At 2 and 16 h after infection, macrophages were lysed by the addition of 0.1% Triton X-100, and serial dilutions were plated onto Mueller-Hinton agar for the enumeration of intracellular bacteria. The intracellular CFU was determined for both time points, and the rates of intracellular replication were expressed as the fold increase in CFU. Experiments were performed in triplicate on three different occasions.

    Article Snippet: S. enterica serotype Typhimurium was detected by using rabbit α- Salmonella O4 test sera (1:1,000; Difco) as a primary antibody and a goat anti-rabbit Cy2-conjugate (1:1,000; Jackson).

    Techniques: Infection, Expressing, Staining

    Host cell phenotypes induced by S. bongori harboring SPI2. (A) Aggregation of host cell endosomes by S. bongori harboring the SPI2 locus and sifA . HeLa cells were infected with S. enterica serotype Typhimurium, S. bongori , or S. bongori harboring the plasmid-borne SPI2 locus. All strains harbored p2812 for the SPI2-dependent expression of sifA ::M45 and the constitutive expression of GFP. At 16 h after infection, cells were fixed and immunostained for the late endosomal-lysosomal membrane protein LAMP-1. Representative micrographs show the organization of endosomal-lysosomal compartments (red) and intracellular Salmonella spp. Salmonella -induced filaments are indicated by arrowheads. (B) Accumulation of host cell tubulin. HeLa cells were infected with S. bongori , S. bongori harboring the plasmid-borne SPI2 locus, or S. bongori harboring the SPI2 locus and plasmid p2643 for the expression of sseF ::HA. HeLa cells were fixed 16 h after infection and immunostaining was performed for S. bongori lipopolysaccharide (green), β-tubulin (red), and translocated SseF-HA (blue). Alternatively, F-actin was visualized by staining with Texas red-phalloidin (red). Representative cells with accumulation of tubulin at the SCV (indicated by arrows) were shown. Such tubulin accumulations were absent in cells infected with S. bongori or S. bongori harboring only plasmid p2643 (not shown). No accumulation of F-actin at the SCV was observed.

    Journal: Infection and Immunity

    Article Title: Functional Transfer of Salmonella Pathogenicity Island 2 to Salmonella bongori and Escherichia coli

    doi: 10.1128/IAI.72.5.2879-2888.2004

    Figure Lengend Snippet: Host cell phenotypes induced by S. bongori harboring SPI2. (A) Aggregation of host cell endosomes by S. bongori harboring the SPI2 locus and sifA . HeLa cells were infected with S. enterica serotype Typhimurium, S. bongori , or S. bongori harboring the plasmid-borne SPI2 locus. All strains harbored p2812 for the SPI2-dependent expression of sifA ::M45 and the constitutive expression of GFP. At 16 h after infection, cells were fixed and immunostained for the late endosomal-lysosomal membrane protein LAMP-1. Representative micrographs show the organization of endosomal-lysosomal compartments (red) and intracellular Salmonella spp. Salmonella -induced filaments are indicated by arrowheads. (B) Accumulation of host cell tubulin. HeLa cells were infected with S. bongori , S. bongori harboring the plasmid-borne SPI2 locus, or S. bongori harboring the SPI2 locus and plasmid p2643 for the expression of sseF ::HA. HeLa cells were fixed 16 h after infection and immunostaining was performed for S. bongori lipopolysaccharide (green), β-tubulin (red), and translocated SseF-HA (blue). Alternatively, F-actin was visualized by staining with Texas red-phalloidin (red). Representative cells with accumulation of tubulin at the SCV (indicated by arrows) were shown. Such tubulin accumulations were absent in cells infected with S. bongori or S. bongori harboring only plasmid p2643 (not shown). No accumulation of F-actin at the SCV was observed.

    Article Snippet: S. enterica serotype Typhimurium was detected by using rabbit α- Salmonella O4 test sera (1:1,000; Difco) as a primary antibody and a goat anti-rabbit Cy2-conjugate (1:1,000; Jackson).

    Techniques: Infection, Plasmid Preparation, Expressing, Immunostaining, Staining

    Expression of sseB by the cloned SPI2 locus. (A) S. enterica serotype Typhimurium wild type (WT), the SPI2 deletion mutant strain (ΔSPI2), and the SPI2 deletion mutant strain complemented with the plasmid-borne SPI2 locus (ΔSPI2 [SPI2]) were grown under conditions resulting in low (LB) or high (PCN-P) levels of the SPI2-encoded protein SseB. After growth for 16 h, lysates were prepared from equal amounts of bacteria, as determined from the OD 600 ). (B) The plasmid-borne SPI2 locus was introduced into S. bongori. S. enterica serotype Typhimurium wild type (S. T.) and S. bongori harboring the vector pBeloBac11 or the SPI2-encoding plasmid pB6 were grown for 16 h in various media as indicated. LB broth was adjusted to pH 7.0 or 5.0, PCN was adjusted to pH 5.8, PCN-P was adjusted to pH 7.4, and F minimal medium containing limiting amounts of Mg 2+ (30 μM) was adjusted to pH 7.0 or 5.0. The amounts of SseB in total bacterial lysates were detected as described above.

    Journal: Infection and Immunity

    Article Title: Functional Transfer of Salmonella Pathogenicity Island 2 to Salmonella bongori and Escherichia coli

    doi: 10.1128/IAI.72.5.2879-2888.2004

    Figure Lengend Snippet: Expression of sseB by the cloned SPI2 locus. (A) S. enterica serotype Typhimurium wild type (WT), the SPI2 deletion mutant strain (ΔSPI2), and the SPI2 deletion mutant strain complemented with the plasmid-borne SPI2 locus (ΔSPI2 [SPI2]) were grown under conditions resulting in low (LB) or high (PCN-P) levels of the SPI2-encoded protein SseB. After growth for 16 h, lysates were prepared from equal amounts of bacteria, as determined from the OD 600 ). (B) The plasmid-borne SPI2 locus was introduced into S. bongori. S. enterica serotype Typhimurium wild type (S. T.) and S. bongori harboring the vector pBeloBac11 or the SPI2-encoding plasmid pB6 were grown for 16 h in various media as indicated. LB broth was adjusted to pH 7.0 or 5.0, PCN was adjusted to pH 5.8, PCN-P was adjusted to pH 7.4, and F minimal medium containing limiting amounts of Mg 2+ (30 μM) was adjusted to pH 7.0 or 5.0. The amounts of SseB in total bacterial lysates were detected as described above.

    Article Snippet: S. enterica serotype Typhimurium was detected by using rabbit α- Salmonella O4 test sera (1:1,000; Difco) as a primary antibody and a goat anti-rabbit Cy2-conjugate (1:1,000; Jackson).

    Techniques: Expressing, Clone Assay, Mutagenesis, Plasmid Preparation

    Translocation of a SPI2 effector protein by intracellular S. enterica serotype Typhimurium and by S. bongori and E. coli harboring the SPI2-encoded type III secretion system. RAW 264.7 cells were grown on glass coverslips and infected with S. enterica serotype Typhimurium, S. bongori , or E. coli Mutaflor. All strains harbored plasmid p2777 for SPI2-regulated expression of sseJ ::HA and constitutive expression of GFP. In addition, S. bongori or E. coli Mutaflor harbored pB6 containing the SPI2 locus, if indicated. At 16 h after infection, the cells were fixed and immunostained for SseJ-HA. Representative micrographs show merged phase contrast, intracellular bacteria (green), and translocated SseJ-HA (red).

    Journal: Infection and Immunity

    Article Title: Functional Transfer of Salmonella Pathogenicity Island 2 to Salmonella bongori and Escherichia coli

    doi: 10.1128/IAI.72.5.2879-2888.2004

    Figure Lengend Snippet: Translocation of a SPI2 effector protein by intracellular S. enterica serotype Typhimurium and by S. bongori and E. coli harboring the SPI2-encoded type III secretion system. RAW 264.7 cells were grown on glass coverslips and infected with S. enterica serotype Typhimurium, S. bongori , or E. coli Mutaflor. All strains harbored plasmid p2777 for SPI2-regulated expression of sseJ ::HA and constitutive expression of GFP. In addition, S. bongori or E. coli Mutaflor harbored pB6 containing the SPI2 locus, if indicated. At 16 h after infection, the cells were fixed and immunostained for SseJ-HA. Representative micrographs show merged phase contrast, intracellular bacteria (green), and translocated SseJ-HA (red).

    Article Snippet: S. enterica serotype Typhimurium was detected by using rabbit α- Salmonella O4 test sera (1:1,000; Difco) as a primary antibody and a goat anti-rabbit Cy2-conjugate (1:1,000; Jackson).

    Techniques: Translocation Assay, Infection, Plasmid Preparation, Expressing

    Rescue of swarming defects with external addition of surfactin and LPS. Five-microliter drops of solution containing surfactin or LPS were placed at the center of 0.5% Difco swarm medium, inoculated with S. enterica serovar Typhimurium ( waaL and cheA ) and E. coli (RP437) strains, and incubated overnight as described in Materials and Methods.

    Journal: Journal of Bacteriology

    Article Title: Genetics of Swarming Motility in Salmonella enterica Serovar Typhimurium: Critical Role for Lipopolysaccharide

    doi:

    Figure Lengend Snippet: Rescue of swarming defects with external addition of surfactin and LPS. Five-microliter drops of solution containing surfactin or LPS were placed at the center of 0.5% Difco swarm medium, inoculated with S. enterica serovar Typhimurium ( waaL and cheA ) and E. coli (RP437) strains, and incubated overnight as described in Materials and Methods.

    Article Snippet: The overall picture emerging from our large-scale search for genes and signals governing swarming motility in S. enterica serovar Typhimurium is that important functions are performed by the LPS layer and the slime and that both known and putative two-component systems play a role in swarming.

    Techniques: Incubation

    Precocious swarming of waaG LPS core mutant. (A) A wild-type (wt) S. enterica serovar Typhimurium strain (SL3770) and its waaG mutant derivative (SL3769) were inoculated on Difco Bacto swarm medium and incubated at 37°C. Photographs were taken after 5 and 20 h of incubation. The arrow points to edge of the waaG swarm colony. (B) The waaG strain was inoculated on minimal swarm medium and incubated at 37°C. Photographs were taken after 24, 30, and 44 h. The arrow indicates edge of the slime ring. Swarming movement was confined to regions that appear as branches emanating from the center.

    Journal: Journal of Bacteriology

    Article Title: Genetics of Swarming Motility in Salmonella enterica Serovar Typhimurium: Critical Role for Lipopolysaccharide

    doi:

    Figure Lengend Snippet: Precocious swarming of waaG LPS core mutant. (A) A wild-type (wt) S. enterica serovar Typhimurium strain (SL3770) and its waaG mutant derivative (SL3769) were inoculated on Difco Bacto swarm medium and incubated at 37°C. Photographs were taken after 5 and 20 h of incubation. The arrow points to edge of the waaG swarm colony. (B) The waaG strain was inoculated on minimal swarm medium and incubated at 37°C. Photographs were taken after 24, 30, and 44 h. The arrow indicates edge of the slime ring. Swarming movement was confined to regions that appear as branches emanating from the center.

    Article Snippet: The overall picture emerging from our large-scale search for genes and signals governing swarming motility in S. enterica serovar Typhimurium is that important functions are performed by the LPS layer and the slime and that both known and putative two-component systems play a role in swarming.

    Techniques: Mutagenesis, Incubation

    Gene expression in S. Typhimurium and C. albicans . (A) Transcription of sopB and sipB of the S. Typhimurium wild type in the presence of supernatants from filaments or yeast-form C. albicans . (B) Transcriptional levels of TEC1 , HWP1 , ALS3 , and CDC42 in

    Journal: Eukaryotic Cell

    Article Title: Killing of Candida albicans Filaments by Salmonella enterica Serovar Typhimurium Is Mediated by sopB Effectors, Parts of a Type III Secretion System ▿

    doi: 10.1128/EC.00014-11

    Figure Lengend Snippet: Gene expression in S. Typhimurium and C. albicans . (A) Transcription of sopB and sipB of the S. Typhimurium wild type in the presence of supernatants from filaments or yeast-form C. albicans . (B) Transcriptional levels of TEC1 , HWP1 , ALS3 , and CDC42 in

    Article Snippet: C. albicans and S. Typhimurium strains were routinely cultured in yeast-peptone-dextrose (YPD) at 30°C and Luria-Bertani (LB) medium (Difco, Detroit, MI) at 37°C, respectively.

    Techniques: Expressing

    S. Typhimurium kills C. albicans filaments via the sopB effector. Viability of C. albicans filaments using the XTT assay. (A) C. albicans Δ tup1 or Δ suv3 strains were cultured at 25°C for 48 h for constitutive filament production

    Journal: Eukaryotic Cell

    Article Title: Killing of Candida albicans Filaments by Salmonella enterica Serovar Typhimurium Is Mediated by sopB Effectors, Parts of a Type III Secretion System ▿

    doi: 10.1128/EC.00014-11

    Figure Lengend Snippet: S. Typhimurium kills C. albicans filaments via the sopB effector. Viability of C. albicans filaments using the XTT assay. (A) C. albicans Δ tup1 or Δ suv3 strains were cultured at 25°C for 48 h for constitutive filament production

    Article Snippet: C. albicans and S. Typhimurium strains were routinely cultured in yeast-peptone-dextrose (YPD) at 30°C and Luria-Bertani (LB) medium (Difco, Detroit, MI) at 37°C, respectively.

    Techniques: XTT Assay, Cell Culture

    The S. Typhimurium sopB effector influences the viability of C. albicans under an in vitro planktonic environment. The viability of C. albicans in the presence of S. Typhimurium wild type (WT) or various mutants was determined at 25°C after 5

    Journal: Eukaryotic Cell

    Article Title: Killing of Candida albicans Filaments by Salmonella enterica Serovar Typhimurium Is Mediated by sopB Effectors, Parts of a Type III Secretion System ▿

    doi: 10.1128/EC.00014-11

    Figure Lengend Snippet: The S. Typhimurium sopB effector influences the viability of C. albicans under an in vitro planktonic environment. The viability of C. albicans in the presence of S. Typhimurium wild type (WT) or various mutants was determined at 25°C after 5

    Article Snippet: C. albicans and S. Typhimurium strains were routinely cultured in yeast-peptone-dextrose (YPD) at 30°C and Luria-Bertani (LB) medium (Difco, Detroit, MI) at 37°C, respectively.

    Techniques: In Vitro

    S. Typhimurium strongly inhibits C. albicans filamentation and biofilm formation via the sopB effector. (A) Attachment of S. Typhimurium wild-type (WT) and sopB mutant strains to the C. albicans filaments. C. albicans SSY50-B filaments were formed in

    Journal: Eukaryotic Cell

    Article Title: Killing of Candida albicans Filaments by Salmonella enterica Serovar Typhimurium Is Mediated by sopB Effectors, Parts of a Type III Secretion System ▿

    doi: 10.1128/EC.00014-11

    Figure Lengend Snippet: S. Typhimurium strongly inhibits C. albicans filamentation and biofilm formation via the sopB effector. (A) Attachment of S. Typhimurium wild-type (WT) and sopB mutant strains to the C. albicans filaments. C. albicans SSY50-B filaments were formed in

    Article Snippet: C. albicans and S. Typhimurium strains were routinely cultured in yeast-peptone-dextrose (YPD) at 30°C and Luria-Bertani (LB) medium (Difco, Detroit, MI) at 37°C, respectively.

    Techniques: Mutagenesis

    Translocation of SopB into C. albicans filaments via SipB. SopB translocation in S. Typhimurium wild-type (top row) and sipB mutant (bottom row) was visualized by confocal microscopy. Scale bars, 4.2 μm. The inset shows an enlarged view of the

    Journal: Eukaryotic Cell

    Article Title: Killing of Candida albicans Filaments by Salmonella enterica Serovar Typhimurium Is Mediated by sopB Effectors, Parts of a Type III Secretion System ▿

    doi: 10.1128/EC.00014-11

    Figure Lengend Snippet: Translocation of SopB into C. albicans filaments via SipB. SopB translocation in S. Typhimurium wild-type (top row) and sipB mutant (bottom row) was visualized by confocal microscopy. Scale bars, 4.2 μm. The inset shows an enlarged view of the

    Article Snippet: C. albicans and S. Typhimurium strains were routinely cultured in yeast-peptone-dextrose (YPD) at 30°C and Luria-Bertani (LB) medium (Difco, Detroit, MI) at 37°C, respectively.

    Techniques: Translocation Assay, Mutagenesis, Confocal Microscopy

    Effect of S. enterica serovar Typhimurium ( S.ty ) porins on Raf-1 (A) and MEK1/2 (B) activation after different stimulation times. Proteins from cell lysates (3 × 10 6 cells/ml) were analyzed by SDS-15% PAGE and immunoblotted with a Raf-1- or MEK1/2-phosphospecific antibody. Shifts in band mobility on SDS-PAGE due to phosphorylation were obtained with anti-Raf-1, anti-MEK1, and anti-MEK2. Fold increases in Raf-1 and MEK1/2 activation, compared to unstimulated cells (control), are shown below each lane for each blot. BSA represents a control of an unspecific stimulus.

    Journal: Infection and Immunity

    Article Title: Porins from Salmonella enterica Serovar Typhimurium Activate the Transcription Factors Activating Protein 1 and NF-?B through the Raf-1-Mitogen-Activated Protein Kinase Cascade

    doi: 10.1128/IAI.70.2.558-568.2002

    Figure Lengend Snippet: Effect of S. enterica serovar Typhimurium ( S.ty ) porins on Raf-1 (A) and MEK1/2 (B) activation after different stimulation times. Proteins from cell lysates (3 × 10 6 cells/ml) were analyzed by SDS-15% PAGE and immunoblotted with a Raf-1- or MEK1/2-phosphospecific antibody. Shifts in band mobility on SDS-PAGE due to phosphorylation were obtained with anti-Raf-1, anti-MEK1, and anti-MEK2. Fold increases in Raf-1 and MEK1/2 activation, compared to unstimulated cells (control), are shown below each lane for each blot. BSA represents a control of an unspecific stimulus.

    Article Snippet: The bacterial strain used was S. enterica serovar Typhimurium SH5014 grown in nutrient broth (Difco, Detroit, Mich.) for 18 to 24 h at 37°C under agitation.

    Techniques: Activation Assay, Polyacrylamide Gel Electrophoresis, SDS Page

    Pattern of S. enterica serovar Typhimurium SH5014 porins and LPS on SDS-PAGE. (A) Lane 1, molecular mass standards (Amersham Pharmacia Biotech, Milan, Italy) (phosphorylase b, 94 kDa,; albumin, 67 kDa; ovalbumin, 43 kDa; carbonic anhydrase, 30 kDa; trypsin inhibitor, 20.1 kDa; α-lactalbumin, 14.4 kDa); lane 2, S. enterica serovar Typhimurium porins (10 μg). (B) Lane 1, S. enterica serovar Typhimurium LPS standard (Sigma-Aldrich S.r.l.); lane 2, S. enterica serovar Typhimurium SH5014 LPS (10 μg).

    Journal: Infection and Immunity

    Article Title: Porins from Salmonella enterica Serovar Typhimurium Activate the Transcription Factors Activating Protein 1 and NF-?B through the Raf-1-Mitogen-Activated Protein Kinase Cascade

    doi: 10.1128/IAI.70.2.558-568.2002

    Figure Lengend Snippet: Pattern of S. enterica serovar Typhimurium SH5014 porins and LPS on SDS-PAGE. (A) Lane 1, molecular mass standards (Amersham Pharmacia Biotech, Milan, Italy) (phosphorylase b, 94 kDa,; albumin, 67 kDa; ovalbumin, 43 kDa; carbonic anhydrase, 30 kDa; trypsin inhibitor, 20.1 kDa; α-lactalbumin, 14.4 kDa); lane 2, S. enterica serovar Typhimurium porins (10 μg). (B) Lane 1, S. enterica serovar Typhimurium LPS standard (Sigma-Aldrich S.r.l.); lane 2, S. enterica serovar Typhimurium SH5014 LPS (10 μg).

    Article Snippet: The bacterial strain used was S. enterica serovar Typhimurium SH5014 grown in nutrient broth (Difco, Detroit, Mich.) for 18 to 24 h at 37°C under agitation.

    Techniques: SDS Page

    Effect of S. enterica serovar Typhimurium ( S.ty ) porins on ERK1/2 (A), p38 (B), and JNK (C) activation after different stimulation times. Proteins from cell lysates (3 × 10 6 cells/ml) were analyzed by SDS-15% PAGE and immunoblotted with an ERK1/2-, p38-, or JNK-phosphospecific antibody. Shifts in band mobility on SDS-PAGE due to phosphorylation were obtained with anti-ERK1/2, anti-p38, and anti-JNK. ERK1 appears as the upper band (44 kDa); ERK2 appears as the lower band (42 kDa). Fold increases in ERK1/2, p38, and JNK activation are shown below each lane for each blot. BSA represents a control of an unspecific stimulus.

    Journal: Infection and Immunity

    Article Title: Porins from Salmonella enterica Serovar Typhimurium Activate the Transcription Factors Activating Protein 1 and NF-?B through the Raf-1-Mitogen-Activated Protein Kinase Cascade

    doi: 10.1128/IAI.70.2.558-568.2002

    Figure Lengend Snippet: Effect of S. enterica serovar Typhimurium ( S.ty ) porins on ERK1/2 (A), p38 (B), and JNK (C) activation after different stimulation times. Proteins from cell lysates (3 × 10 6 cells/ml) were analyzed by SDS-15% PAGE and immunoblotted with an ERK1/2-, p38-, or JNK-phosphospecific antibody. Shifts in band mobility on SDS-PAGE due to phosphorylation were obtained with anti-ERK1/2, anti-p38, and anti-JNK. ERK1 appears as the upper band (44 kDa); ERK2 appears as the lower band (42 kDa). Fold increases in ERK1/2, p38, and JNK activation are shown below each lane for each blot. BSA represents a control of an unspecific stimulus.

    Article Snippet: The bacterial strain used was S. enterica serovar Typhimurium SH5014 grown in nutrient broth (Difco, Detroit, Mich.) for 18 to 24 h at 37°C under agitation.

    Techniques: Activation Assay, Polyacrylamide Gel Electrophoresis, SDS Page

    Western blotting analysis with pJNK and phospho-p38 antibodies of lysates from C3H/HeJ peritoneal macrophages (5 × 10 6 cells/ml) after stimulation with LPS (1 μg/ml) or porins (5 μg/ml) for 20 min. The experimental procedure was as described in Materials and Methods. S. ty ., S. enterica serovar Typhimurium.

    Journal: Infection and Immunity

    Article Title: Porins from Salmonella enterica Serovar Typhimurium Activate the Transcription Factors Activating Protein 1 and NF-?B through the Raf-1-Mitogen-Activated Protein Kinase Cascade

    doi: 10.1128/IAI.70.2.558-568.2002

    Figure Lengend Snippet: Western blotting analysis with pJNK and phospho-p38 antibodies of lysates from C3H/HeJ peritoneal macrophages (5 × 10 6 cells/ml) after stimulation with LPS (1 μg/ml) or porins (5 μg/ml) for 20 min. The experimental procedure was as described in Materials and Methods. S. ty ., S. enterica serovar Typhimurium.

    Article Snippet: The bacterial strain used was S. enterica serovar Typhimurium SH5014 grown in nutrient broth (Difco, Detroit, Mich.) for 18 to 24 h at 37°C under agitation.

    Techniques: Western Blot