Journal: Journal of Bacteriology

Article Title: Growth- and Stress-Induced PASTA Kinase Phosphorylation in Enterococcus faecalis

doi: 10.1128/JB.00363-17

Figure Lengend Snippet: Stimulation of IreK phosphorylation and activity by antimicrobial-induced cell wall stress. Exponentially growing wild-type E. faecalis OG1 cells were harvested with or without treatment with ceftriaxone (128 μg/ml) for 20 min or were suspended in PBS prior to treatment with ceftriaxone or lysozyme (50 μg/ml). Whole-cell lysates were subjected to Phos-tag SDS-PAGE and analyzed as for Fig. 2. Immunoblotting with specific antisera was used to detect IreK (A) or IreB (B). Asterisks indicate that changes in phosphorylation were statistically significant (P < 0.05) when bracketed bars were compared, using Student's t test (2-tailed, heteroscedastic). Exp, exponentially growing cells; PBS, cells suspended in PBS; UT, untreated; Cftx, ceftriaxone treated; Lys, lysozyme treated.

Article Snippet: When required, antimicrobials were used at the following concentrations unless otherwise noted: ceftriaxone, 128 μg/ml (GoldBio); chloramphenicol, 8 μg/ml (Sigma); erythromycin, 4 μg/ml (GoldBio); lysozyme, 50 μg/ml (Sigma); vancomycin, 4 μg/ml (GoldBio); bacitracin, 128 μg/ml (Sigma); fosfomycin, 128 μg/ml (Sigma); rifampin, 16 μg/ml (Sigma).

Techniques: Activity Assay, SDS Page, Western Blot

Journal: Journal of Bacteriology

Article Title: Growth- and Stress-Induced PASTA Kinase Phosphorylation in Enterococcus faecalis

doi: 10.1128/JB.00363-17

Figure Lengend Snippet: Requirement for residues in the IreK activation loop for IreK phosphorylation and activity. Exponentially growing E. faecalis cells exposed to ceftriaxone (128 μg/ml) for 20 min (Cftx) or untreated controls (UT) were harvested. Whole-cell lysates were subjected to Phos-tag SDS-PAGE and analyzed as for Fig. 2. Immunoblotting with specific antisera was used to detect IreK (A) or IreB (B). Asterisks indicate that changes in phosphorylation were statistically significant (P < 0.05) when bracketed bars were compared, using Student's t test (2-tailed, heteroscedastic). The strains used were wild-type (WT) OG1, OK5 (ireK T163A T166A T168A) (T->A), and YC19 (ireK T163E T166E T168E) (T->E).

Article Snippet: When required, antimicrobials were used at the following concentrations unless otherwise noted: ceftriaxone, 128 μg/ml (GoldBio); chloramphenicol, 8 μg/ml (Sigma); erythromycin, 4 μg/ml (GoldBio); lysozyme, 50 μg/ml (Sigma); vancomycin, 4 μg/ml (GoldBio); bacitracin, 128 μg/ml (Sigma); fosfomycin, 128 μg/ml (Sigma); rifampin, 16 μg/ml (Sigma).

Techniques: Activation Assay, Activity Assay, SDS Page, Western Blot

Journal: Journal of Bacteriology

Article Title: Growth- and Stress-Induced PASTA Kinase Phosphorylation in Enterococcus faecalis

doi: 10.1128/JB.00363-17

Figure Lengend Snippet: Requirement for the PASTA module for activation of IreK in response to cell wall stress but not for activation driven by growth and/or cell division. Whole-cell lysates from exponentially growing treated or untreated cells were subjected to Phos-tag SDS-PAGE and analyzed as for Fig. 2. Asterisks indicate statistically significant (P < 0.05) changes when bracketed bars were compared, using Student's t test (2-tailed, heteroscedastic). (A) Analysis of an E. faecalis mutant (CK212) in which all five PASTA domains were deleted from IreK. (B) Analysis of growth-induced signaling and lysozyme responses in CK212 cells. (C) Analysis of cell envelope integrity by CPRG hydrolysis. The strains (carrying pCJK205) were wild-type OG1, JL206 (ΔireK), BL102 (ireK K41R), and CK212 (ireK ΔPASTA1–5). Asterisks indicate statistically significant (P < 0.05) changes, compared to the wild-type strain, using Student's t test (2-tailed, heteroscedastic). Error bars indicate 1 standard deviation. (D) Analysis of phosphorylation in E. faecalis ireK mutants carrying deletions of individual PASTA domains. The strains were as follows: wild type, OG1; ΔP1, BL94; ΔP2, BL95; ΔP3, BL96; ΔP4, BL97; ΔP5, BL98. (E) Analysis of phosphorylation in E. faecalis ireK mutants carrying C-terminal truncations of PASTA domains. The strains were as follows: wild type, OG1; ΔP5, BL98; ΔP4–5, BL104; ΔP3–5, BL106; ΔP2–5, BL111; ΔP1–5, CK212. Exp, exponentially growing cells; PBS, cells suspended in PBS; UT, untreated; Cftx, ceftriaxone treated; Lys, lysozyme.

Article Snippet: When required, antimicrobials were used at the following concentrations unless otherwise noted: ceftriaxone, 128 μg/ml (GoldBio); chloramphenicol, 8 μg/ml (Sigma); erythromycin, 4 μg/ml (GoldBio); lysozyme, 50 μg/ml (Sigma); vancomycin, 4 μg/ml (GoldBio); bacitracin, 128 μg/ml (Sigma); fosfomycin, 128 μg/ml (Sigma); rifampin, 16 μg/ml (Sigma).

Techniques: Activation Assay, SDS Page, Mutagenesis, Standard Deviation

Journal: Neurobiology of aging

Article Title: Ceftriaxone ameliorates tau pathology and cognitive decline via restoration of glial glutamate transporter in a mouse model of Alzheimer's disease.

doi: 10.1016/j.neurobiolaging.2015.04.005

Figure Lengend Snippet: Fig. 3. Chronic ceftriaxone-treatment increases GLT-1 expression levels in 3xTg-Alzheimer’s disease (AD) mice. Western blot analysis of protein extracts from hippocampal tissue of 12-month-old 3xTg-AD mice using GLT-1 and GAPDH (housekeeping) antibodies. Densitometric analysis of the GLT-1 bands normalized to GAPDH shown in the graph. Each bar is expressed as mean standard error of the mean; *p < 0.05, ceftriaxone-treated 3xTg-AD mice compared with the vehicle-treated 3xTg-AD mice (n ¼ 5 mice per group). Molecular weight markers are in kDa. Abbreviations: Cef, ceftriaxone treatment; GLT-1, glutamate transporter 1; Veh, vehicle or saline-treatment.

Article Snippet: Ten-month-old 3xTg-AD mice were treated intraperitoneally (i.p.) with 200 mg/kg ceftriaxone (Santa Cruz Biotechnology, Santa Cruz, CA, USA) or 0.8% sodium chloride (Vehicle) daily for 2 months (Rothstein et al., 2005).

Techniques: Expressing, Western Blot, Molecular Weight, Saline

Journal: Neurobiology of aging

Article Title: Ceftriaxone ameliorates tau pathology and cognitive decline via restoration of glial glutamate transporter in a mouse model of Alzheimer's disease.

doi: 10.1016/j.neurobiolaging.2015.04.005

Figure Lengend Snippet: Fig. 4. Upregulation of GLT-1 by ceftriaxone ameliorates cognitive decline in 3xTg-AD mice. (A) Acquisition curve during training of Morris water maze (MWM) expressed as average of all mice in each group. Arrowheads show the first training of the day. (B) Escape latency and (C) number of crosses on platform location for 24-hour retention trial in MWM. (D) Novel object recognition test for the first minute of the test. Each bar is expressed as mean standard error of the mean; *p < 0.05 and **p < 0.01, ceftriaxone-treated 3xTg-AD mice compared with the vehicle-treated 3xTg-AD mice (n ¼ 5 mice per group). Abbreviation: GLT-1, glutamate transporter 1.

Article Snippet: Ten-month-old 3xTg-AD mice were treated intraperitoneally (i.p.) with 200 mg/kg ceftriaxone (Santa Cruz Biotechnology, Santa Cruz, CA, USA) or 0.8% sodium chloride (Vehicle) daily for 2 months (Rothstein et al., 2005).

Techniques:

Journal: Neurobiology of aging

Article Title: Ceftriaxone ameliorates tau pathology and cognitive decline via restoration of glial glutamate transporter in a mouse model of Alzheimer's disease.

doi: 10.1016/j.neurobiolaging.2015.04.005

Figure Lengend Snippet: Fig. 5. Effect of GLT-1 upregulation on Ab pathology in 3xTg-AD mice. (A) Quantitative Ab ezyme-linked immunosorbent assay in detergent-insoluble (formic acid soluble) brain hippocampal fraction and detergent-soluble brain hippocampal fraction. (B) Western blot analysis of APP processing in the hippocampus of the brain. The densitometric analysis of C99 and C83 fragments shown in the graph. (C) Dot blot analysis of soluble prefibrillar and fibrillar Ab oligomers in the cortical soluble fractions detected by the A11 and OC antibodies respectively. (D) Qualitative representation of the immunohistochemical staining with 4G8 and thioflavin S to detect Ab plaques. Each bar is expressed in mean standard error of the mean; **p < 0.01, ceftriaxone-treated 3xTg-AD mice compared with the vehicle-treated 3xTg-AD mice (n ¼ 4e5 mice per group). Abbreviations: Ab, amyloid- beta; AD, Alzheimer’s disease; Cef, ceftriaxone; GLT-1, glutamate transporter 1; Veh, vehicle.

Article Snippet: Ten-month-old 3xTg-AD mice were treated intraperitoneally (i.p.) with 200 mg/kg ceftriaxone (Santa Cruz Biotechnology, Santa Cruz, CA, USA) or 0.8% sodium chloride (Vehicle) daily for 2 months (Rothstein et al., 2005).

Techniques: Western Blot, Dot Blot, Immunohistochemical staining, Staining

Journal: Neurobiology of aging

Article Title: Ceftriaxone ameliorates tau pathology and cognitive decline via restoration of glial glutamate transporter in a mouse model of Alzheimer's disease.

doi: 10.1016/j.neurobiolaging.2015.04.005

Figure Lengend Snippet: Fig. 6. Effect of GLT-1 upregulation on tau pathology in 3xTg-AD mice. (A) Representative Immunohistochemical or immunofluorescence staining with tau antibodies HT7, PHF-1, and CP13 antibodies. The mean intensities of HT7 and PHF-1 from immunofluorescence staining are shown in the graph along with the CP13 positive neuron analysis. Arrows point to tau-positive neurons. (B) Representative Western blot image of total human tau using HT7 antibody from the sarkosyleinsoluble cortical protein fraction. The densitometric analysis of all bands are shown in the graph. (C) Western blot analysis of total human tau using HT7 antibody and various phosphor-specific tau using PHF1, AT8, CP13, AT180, and GAPDH (housekeeping protein) from the detergent-soluble hippocampal protein fraction. The densitometric analysis of PHF1, AT8, CP13 and AT180 are shown in the graph. (D) Western blot analysis of known tau kinases: p25, p35, phosphor-GSK3b (pGSK3b), GSK3b, and tubulin (housekeeping protein) on detergent-soluble hippocampal protein fraction. The densitometric analysis of p25, p35, phosphor-GSK3b, and GSK3b are shown in the graph. Each bar is expressed as mean standard error of the mean; *p < 0.05, ceftriaxone- treated 3xTg-AD mice compared with the vehicle-treated 3xTg-AD mice (n ¼ 4e5 mice per group); ****p < 0.0001. Abbreviations: AD, Alzheimer’s disease; Cef, ceftriaxone; GLT-1, glutamate transporter 1; Veh, vehicle.

Article Snippet: Ten-month-old 3xTg-AD mice were treated intraperitoneally (i.p.) with 200 mg/kg ceftriaxone (Santa Cruz Biotechnology, Santa Cruz, CA, USA) or 0.8% sodium chloride (Vehicle) daily for 2 months (Rothstein et al., 2005).

Techniques: Immunohistochemical staining, Staining, Western Blot

Journal: Neurobiology of aging

Article Title: Ceftriaxone ameliorates tau pathology and cognitive decline via restoration of glial glutamate transporter in a mouse model of Alzheimer's disease.

doi: 10.1016/j.neurobiolaging.2015.04.005

Figure Lengend Snippet: Fig. 7. Synaptic proteins restored in ceftriaxone-treated 3xTg-Alzheimer’s disease (AD) mice. (A) Representative immunofluorescence (IF) images of SYP in the CA3 region of the hippocampus and quantitative analysis of the average mean intensities from 5 IF (20 mm) stained sections in the dentate gyrus (DG), CA1, and CA3 of the hippocampus. (B) Representative IF images of PSD95 in the CA3 region of the hippocampus and quantitative analysis of the average mean intensities from 5 IF (20 mm) stained sections in the DG, CA1, and CA3 of the hippocampus. (C) Escape latency as a function of synaptic protein mean intensity graphs in the designated region of the hippocampus. Each shape represents 1 mouse and each bar is expressed as mean standard error of the mean; *p < 0.05 and ***p < 0.001, ceftriaxone-treated 3xTg-AD mice compared with the vehicle-treated 3xTg-AD mice (n ¼ 3e5 mice per group). Abbreviations: Cef, ceftriaxone; PSD95, postsynaptic density protein 95 kDa; sec, seconds; SYP, synaptophysin; Veh, vehicle.

Article Snippet: Ten-month-old 3xTg-AD mice were treated intraperitoneally (i.p.) with 200 mg/kg ceftriaxone (Santa Cruz Biotechnology, Santa Cruz, CA, USA) or 0.8% sodium chloride (Vehicle) daily for 2 months (Rothstein et al., 2005).

Techniques: Staining

Journal: MicrobiologyOpen

Article Title: Evaluating Effects of Antibiotics Across Preclinical Models of the Human Gastrointestinal Microbiota

doi: 10.1002/mbo3.70030

Figure Lengend Snippet: Partial recovery of microbial diversity following treatment with a subset of antibiotics for human fecal communities cultured in MBRAs. (A) Observed OTUs assessed two days following the end of treatment (Recovery, Day 7) were plotted along with data collected before (Day 0) and after (Day 5) treatment with the treatment indicated on the x ‐axis. Individual points were plotted from the mean of replicates for each of the 12 fecal samples tested. (B) Jaccard and (C) Bray‐Curtis dissimilarity values from samples before treatment (Day 0) were determined for samples collected after treatment (Day 0–Day 5) or after a two‐day recovery following antibiotic treatment (Day 0–Day 7), with the treatment indicated on the x‐axis. Individual points were plotted from the mean of replicate dissimilarity measures for each of the 12 fecal samples tested. In all panels, data from before and after treatment were reproduced from Figure , with the exception of data from cefaclor and ceftriaxone‐treated communities, which were restricted to duplicate samples collected from reactors where community composition was measured on Day 7. Median values were plotted as horizontal lines, interquartile (IQR) ranges were plotted as boxes, and range of data was plotted as whiskers. Statistical significance was determined by paired ANOVA with Geisser‐Greenhouse correction for unequal variability of differences and Holm‐Šidák correction for multiple comparisons. In (A), comparisons were performed between all time points from the same treatment. In (B) and (C), comparisons were made between dissimilarity values for the same treatment. ns > 0.05, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: Ceftriaxone sodium salt hemiheptahydrate , Thermo Scientific Chemicals.

Techniques: Cell Culture

Journal: MicrobiologyOpen

Article Title: Evaluating Effects of Antibiotics Across Preclinical Models of the Human Gastrointestinal Microbiota

doi: 10.1002/mbo3.70030

Figure Lengend Snippet: Levels of acetate and butyrate were lower in clindamycin‐treated communities. We measured levels of (A) acetate, (B) butyrate, and (C) isovalerate in filter‐sterilized spent culture medium from MBRA communities on Day 5 in untreated ( n = 24) and antibiotic‐treated communities ( n = 12, except cefaclor and ceftriaxone, where n = 24). Statistical significance of differences between antibiotic‐treated and untreated communities was determined with one‐way ANOVA with Brown‐Forsythe and Welch correction for unequal variances and Dunnett's correction for multiple comparisons. * p < 0.05, ** p < 0.01. Comparisons to untreated with p > 0.05 are not shown.

Article Snippet: Ceftriaxone sodium salt hemiheptahydrate , Thermo Scientific Chemicals.

Techniques:

Journal: BMC Cancer

Article Title: Cholesterol-induced colorectal cancer progression and its mitigation through gut microbiota remodeling and simvastatin treatment

doi: 10.1186/s12885-025-14379-3

Figure Lengend Snippet: Lactobacillus plantarum-mediated reduction of serum cholesterol alleviates CRC tumor growth. ( A ) In vivo bioluminescence imaging showing tumor progression in four groups: I (Lactobacillus plantarum + Normal diet), II (Normal diet), III (Antibiotics), and IV (Antibiotics + Lactobacillus plantarum). Antibiotics group: 0.02mL/g ceftriaxone intragastric administration every two days. ( B ) Quantification of fluorescence intensity representing tumor size across the four groups. ( C ) Serum cholesterol and LDL/VLDL expression levels (µg/µl), showing a significant reduction with Lactobacillus plantarum supplementation. ( D ) Histological analysis of colon tissues from each group, with HE staining showing tissue architecture and Ki-67 staining highlighting proliferating cells. ( E - G ) Relative expression levels of inflammatory markers IL-2, TNF-α, and IFN-γ, respectively, in colon tissues across the four groups. Data are presented as mean ± SEM, with significant differences indicated (*, P < 0.05; **, P < 0.01; ***, P < 0.001)

Article Snippet: In the antibiotic depletion group, the model mice were given 0.02 mL/g ceftriaxone intragastric administration every two days, and the concentration of ceftriaxone sodium was 0.6 g/mL (MCE, HY-B0712).

Techniques: In Vivo, Imaging, Fluorescence, Expressing, Staining

Journal: Antibiotics

Article Title: Mapping Escherichia coli in Women with Simple Urinary Tract Infections: Phenotypic ESBL/AmpC Screening and Whole-Genome Insights from Oman

doi: 10.3390/antibiotics15020124

Figure Lengend Snippet: Double Disc Synergy Test (DDST). Detection of ESBL production in E. coli using several cephalosporins: cefepime (FEP), Cefpodoxime (PX), Ceftriaxone (CRO), Cefixime (CFM), and Cefaclor (CEC) against two different β-lactam/β-lactamase inhibitors: Amoxicillin-clavulanate (AMC) and Piperacillin/tazobactam (TZP). Black arrows indicate some synergistic pattern in the DDST.

Article Snippet: The BL/BLIs were placed at the center of the Mueller Hinton agar plates and cefepime (FEP, 30 μg) (Oxoid, Basingstoke, UK), cefixime (CFM, 5 μg) (Oxoid, Basingstoke, UK), cefuroxime (CXM, 30 μg) (Oxoid, Basingstoke, UK), cefazolin (KZ, 30 μg) (Oxoid, Basingstoke, UK), cefaclor (CEC, 30 μg) (Oxoid, Basingstoke, UK), and ceftriaxone (CRO, 30 μg) (Oxoid, Basingstoke, UK) were placed at a distance of 20 mm edge to edge from them.

Techniques: