Journal: Antibiotics

Article Title: Limosilactobacillus fermentum Strain 3872: Antibacterial and Immunoregulatory Properties and Synergy with Prebiotics against Socially Significant Antibiotic-Resistant Infections of Animals and Humans

doi: 10.3390/antibiotics11101437

Figure Lengend Snippet: Antagonistic activity of LF3872 against E. coli strains.

Article Snippet: E. coli ATCC E 2348/69 , 41.5 ± 3.7 , 64.5 ± 5.8 *.

Techniques: Activity Assay

Journal: Antibiotics

Article Title: Limosilactobacillus fermentum Strain 3872: Antibacterial and Immunoregulatory Properties and Synergy with Prebiotics against Socially Significant Antibiotic-Resistant Infections of Animals and Humans

doi: 10.3390/antibiotics11101437

Figure Lengend Snippet: Extracellular ATP levels in tested bacteria treated with CSLF3872.

Article Snippet: E. coli ATCC E 2348/69 , 41.5 ± 3.7 , 64.5 ± 5.8 *.

Techniques: Bacteria

Journal: Antibiotics

Article Title: Limosilactobacillus fermentum Strain 3872: Antibacterial and Immunoregulatory Properties and Synergy with Prebiotics against Socially Significant Antibiotic-Resistant Infections of Animals and Humans

doi: 10.3390/antibiotics11101437

Figure Lengend Snippet: Co-aggregative abilities of LF3872 strain with proteobacteria pathogens.

Article Snippet: E. coli ATCC E 2348/69 , 41.5 ± 3.7 , 64.5 ± 5.8 *.

Techniques:

Journal: Antibiotics

Article Title: Limosilactobacillus fermentum Strain 3872: Antibacterial and Immunoregulatory Properties and Synergy with Prebiotics against Socially Significant Antibiotic-Resistant Infections of Animals and Humans

doi: 10.3390/antibiotics11101437

Figure Lengend Snippet: Synergism of CSLF3872 and Actigen prebiotic in inhibiting the adhesion of proteobacteria pathogens ( E. coli , Salmonella , Campylobacter ) to Caco-2 cells.

Article Snippet: E. coli ATCC E 2348/69 , 41.5 ± 3.7 , 64.5 ± 5.8 *.

Techniques: Control

Journal: Antibiotics

Article Title: Limosilactobacillus fermentum Strain 3872: Antibacterial and Immunoregulatory Properties and Synergy with Prebiotics against Socially Significant Antibiotic-Resistant Infections of Animals and Humans

doi: 10.3390/antibiotics11101437

Figure Lengend Snippet: Microorganisms used in this study.

Article Snippet: E. coli ATCC E 2348/69 , 41.5 ± 3.7 , 64.5 ± 5.8 *.

Techniques:

Journal: PLoS Pathogens

Article Title: Unique features in the intracellular transport of typhoid toxin revealed by a genome-wide screen

doi: 10.1371/journal.ppat.1007704

Figure Lengend Snippet: (A) Genotyping of the CRISPR/Cas9-generated knockout cells. Genomic DNA was purified from CRISPR-Cas9 edited HEK293T cell lines and analyzed by PCR with specific primers listed in . The TMED2-deficient cell line was examined by Western blot with an anti TEMED2-specific antibody. Dotted lines indicate places where the experimentally relevant lanes were spliced together (all lanes originate from a single gel). (B) Relative toxicity of typhoid toxin in the indicated knockout cell lines after treatment with a serial dilution of purified toxin. The relative toxicity was determined from the percentage of cells at the G 2 M phase fitted by nonlinear regression as indicated in the Materials and Methods. Values, which were normalized relative to wild type (considered 100) are the mean ± SEM of independent determinations. ****p < 0.0001; n. s.: differences not statistically significant; two-tailed Student’s t-test.

Article Snippet: Antibodies to Myc (Cell Signaling Technology, Cat. #2276), TMED2 (Santa Cruz Biotechnology, Cat.# sc376458), and GM130 (BD Bioscience, Cat. # 610822) were purchased from the indicated commercial sources.

Techniques: CRISPR, Generated, Knock-Out, Purification, Western Blot, Serial Dilution, Two Tailed Test

Journal: Journal of Biological Chemistry

Article Title: Protection of telomeres 1 proteins POT1a and POT1b can repress ATR signaling by RPA exclusion, but binding to CST limits ATR repression by POT1b

doi: 10.1074/jbc.ra118.004598

Figure Lengend Snippet: Figure 3. ATR repression by POT1b is limited by its interaction with CST. A, schematic of the role of POT1b in recruiting CST and polymerase /primase to telomeres (top). Schematic of POT1b domains and the mutations that disrupt the POT1b-CST interaction (bottom). B, immunoblot for POT1b in POT1aF/F MEFs expressing the indicated proteins before and after treatment with Cre (96 h). ctrl, nonspecific band used as loading control. C, representative IF-FISH images showing the DNA damage response at telomeres in Cre-treated POT1aF/F MEFs expressing POT1a, POT1b, or POT1bCST (96 h). H2AX foci that co-localize with telomeres (TIFs) were detected using -H2AX antibody (red) and a telomeric FISH probe (green). D, quantification of the TIF response as shown in C of POT1aF/F MEFs before and after Cre treatment. Bar graphs represent averages from three experiments and S.D.s. p values were derived from a two-tailed Student’s t test. p values: **, p 0.01; *, p 0.05. E, immunoblot for POT1a, POT1b, and Stn1 in POT1aF/F MEFs (with and without 96 h Cre treatment) expressing exogenous POT1b and/or Stn1 shRNA (indicated with ) or a scrambled sh (indicated with ). Asterisks indicate nonspecific bands. ctrl, nonspecific band used as loading control. F, representative images of IF-FISH to monitor the DNA damage response at telomeres in the indicated Cre-treated (96 h) POT1aF/F MEFs with and without POT1b and expressing Stn1sh or a scrambled control (ctrl sh). H2AX foci that co-localize with telomeres (TIFs) were detected with IF with an -H2AX antibody (red) and FISH with a telomeric PNA probe (green). G, quantification of TIF response in the indicated POT1aF/F MEFs (as in F). Bar graphs represent averages from three independent experiments and S.D.s. p values: **, p 0.001; *, p 0.05; ns, not significant.

Article Snippet: After immunoblotting, the membranes were blocked in TBS with Tween 20 with 2.5% nonfat dry milk and incubated with the following primary antibodies -POT1a (no. 1221), -Myc (9B11, Cell Signaling Technology), -TIN2 (no. 1447), -Rap1 (no. 1253), -RPA32 (A300 –244A, Bethyl), -RPA70 (A300 –241A, Bethyl), -POT1b (no. 1223), -STN1 (sc376450, Santa Cruz Biotechnology), -tubulin (GTU88, Abcam) in 1% nonfat dry milk.

Techniques: Western Blot, Expressing, Control, Derivative Assay, Two Tailed Test, shRNA

Journal: The Journal of Biological Chemistry

Article Title: Sterol carrier protein-2 deficiency attenuates diet-induced dyslipidemia and atherosclerosis in mice

doi: 10.1074/jbc.RA118.002290

Figure Lengend Snippet: SCP2 deficiency significantly attenuates diet-induced atherosclerosis. At 10 weeks of age, LDLR−/− and LS mice were fed a high-fat, high-cholesterol–containing Western-type diet (TD88137) for 16 weeks. Aortae were dissected and prepared for en face analyses. The total area as well as the area occupied by the lesions was quantified using Axiovision software and expressed as percent lesion area. A, representative images. B, percent area occupied by the lesions in the aortic arch. C, percent area occupied by the lesions in the entire aorta. *, p < 0.05; individual p values are included in the text.

Article Snippet: Cells were transfected with a complex consisting of SCP2 siRNA (Santa Cruz Biotechnology, catalog no. sc-44636) and polyamidoamine (PAMAM) G5 (Sigma-Aldrich).

Techniques: Western Blot, Software

Journal: The Journal of Biological Chemistry

Article Title: Sterol carrier protein-2 deficiency attenuates diet-induced dyslipidemia and atherosclerosis in mice

doi: 10.1074/jbc.RA118.002290

Figure Lengend Snippet: SCP2 deficiency reduces plaque size as well as plaque necrosis in the aortic root. Paraffin-embedded aortic root sections (5 μm) were stained with H&E or Masson's trichrome, imaged, and analyzed by Axiovision software. A, representative H&E stained images of the aortic root of the indicated genotypes/sex. Scale bar = 100 μm. B, the total aortic root and area occupied by the lesions was quantified, and data (mean ± S.D., n = 6) are presented as percent lesion area. C, representative trichrome-stained images of the indicated genotypes/sex. Scale bar = 50 μm. D, the total and necrotic areas were quantified for all three aortic valve leaflets, and data (mean ± S.D., n = 12 leaflets) are presented as percent necrotic area. *, p < 0.05.

Article Snippet: Cells were transfected with a complex consisting of SCP2 siRNA (Santa Cruz Biotechnology, catalog no. sc-44636) and polyamidoamine (PAMAM) G5 (Sigma-Aldrich).

Techniques: Staining, Software

Journal: The Journal of Biological Chemistry

Article Title: Sterol carrier protein-2 deficiency attenuates diet-induced dyslipidemia and atherosclerosis in mice

doi: 10.1074/jbc.RA118.002290

Figure Lengend Snippet: SCP2 deficiency significantly reduces plasma cholesterol and triglyceride levels. At 10 weeks of age, LDLR−/− and LS mice were fed a high-fat, high-cholesterol–containing Western-type diet (TD88137) for 16 weeks. After an overnight fast, mice were euthanized, and fasting plasma was collected. A, levels of total plasma cholesterol for indicated genotype and sexes. B, percent of total plasma cholesterol associated with the non-HDL or HDL fraction. C, percent of total plasma cholesterol associated with the non-HDL or HDL fraction in both genotypes normalized to total plasma cholesterol in LDLR−/− mice of the respective sex. D, linear regression analyses of total lesion area and plasma cholesterol; the observed coefficient of correlation (R) as significance of correlation (p value) is indicated. E, total plasma triglyceride for the indicated genotype and sexes. F, linear regression analyses of total lesion area and plasma triglyceride levels; the observed coefficients of correlation (R) as significance of correlation (p value) is indicated.

Article Snippet: Cells were transfected with a complex consisting of SCP2 siRNA (Santa Cruz Biotechnology, catalog no. sc-44636) and polyamidoamine (PAMAM) G5 (Sigma-Aldrich).

Techniques: Clinical Proteomics, Western Blot

Journal: The Journal of Biological Chemistry

Article Title: Sterol carrier protein-2 deficiency attenuates diet-induced dyslipidemia and atherosclerosis in mice

doi: 10.1074/jbc.RA118.002290

Figure Lengend Snippet: SCP2 deficiency significantly reduces intestinal cholesterol absorption, and SCP2 knockdown in intestinal epithelial cells reduces cholesterol uptake. A, C57BL/6 (WT) or SCP2−/− mice were injected intravenously with the lipoprotein lipase inhibitor tyloxapol (500 mg/kg body weight) and gavaged with [3H]cholesterol in olive oil (2 μCi in 200 μl) after 5 min. Intestinal cholesterol absorption was assessed by monitoring the radiolabel associated with plasma collected at the time of euthanasia. Data are presented as plasma DPM per microliter of plasma for the indicated genotype and sex. B, the entire length of the intestine, from the base of the stomach to the tip of the cecum, was divided into four equal parts (P1 to P4), and total RNA was isolated. The mRNA levels of indicated genes were determined by real-time PCR as described under “Experimental procedures” and normalized to the housekeeping gene β-actin. C, total protein extracts from ileum segments P1 to P4 of WT and SCP2−/− mice as well as the colon (C) were analyzed by Western blotting for expression of SCP2; β-actin was used as the loading control. Human intestinal epithelial cells (HT-29) were transfected with scrambled or SCP2-specific siRNA as described under “Experimental procedures.” Total protein or RNA was extracted and used to determine the levels of SCP2. D, top, a representative Western blot. Bottom, levels of SCP2 mRNA quantified by quantitative PCR and SCP2 protein by densitometric analyses of Western blots. Data are presented as percent scrambled siRNA-transfected controls for the indicated SCP2 siRNA concentrations used. E, HT-29 cells transfected with scrambled siRNA or 53.3 nm SCP2-specific siRNA were incubated with [3H]-cholesterol, and total cellular uptake was monitored as described under “Experimental procedures.” Data (mean ± S.D., n = 6) are presented as DPM normalized to total cellular protein. *, p < 0.05.

Article Snippet: Cells were transfected with a complex consisting of SCP2 siRNA (Santa Cruz Biotechnology, catalog no. sc-44636) and polyamidoamine (PAMAM) G5 (Sigma-Aldrich).

Techniques: Knockdown, Injection, Clinical Proteomics, Isolation, Real-time Polymerase Chain Reaction, Western Blot, Expressing, Control, Transfection, Incubation

Journal: The Journal of Biological Chemistry

Article Title: Sterol carrier protein-2 deficiency attenuates diet-induced dyslipidemia and atherosclerosis in mice

doi: 10.1074/jbc.RA118.002290

Figure Lengend Snippet: SCP2 deficiency significantly reduces lipid secretion from liver and isolated hepatocytes. A, C57BL/6 (WT) or SCP2−/− mice were injected with the lipoprotein lipase inhibitor tyloxapol (500 mg/kg of body weight), and blood samples were drawn at 0 and 3 h. Triglyceride secretion rates for indicated genotypes and sexes are presented. B and C, primary hepatocytes were prepared from C57BL/6 (WT) or SCP2−/− mice. Following incubation with [3H]oleic acid, radiolabel associated with secreted triglycerides (TG, B) or cholesteryl esters (C) was determined as described under “Experimental procedures” and normalized to cellular protein. Data are presented as DPM associated with the triglyceride or cholesteryl ester fraction in the total lipids extracted from the medium per milligram of total protein.

Article Snippet: Cells were transfected with a complex consisting of SCP2 siRNA (Santa Cruz Biotechnology, catalog no. sc-44636) and polyamidoamine (PAMAM) G5 (Sigma-Aldrich).

Techniques: Isolation, Injection, Incubation

Journal: The Journal of Biological Chemistry

Article Title: Sterol carrier protein-2 deficiency attenuates diet-induced dyslipidemia and atherosclerosis in mice

doi: 10.1074/jbc.RA118.002290

Figure Lengend Snippet: SCP2 deficiency leads to reduced lipid accumulation in the liver without a change in the expression of lipogenic genes. A, liver tissue harvested from WD-fed LDLR−/− or LS mice were paraffin-embedded, and 5-μm sections were stained with H&E. Images were acquired using a Zeiss inverted microscope fitted with a digital camera. Scale bar = 50 μm. B, SCP2 mRNA levels in total liver RNA were determined by real-time PCR as described under “Experimental procedures” and normalized to the housekeeping gene β-actin. Relative expression (mean ± S.D., n = 6) is shown. C, hepatic mRNA levels of the indicated genes were determined by real-time PCR as described under “Experimental procedures” and normalized to the housekeeping gene β-actin. Relative expression (mean ± S.D., n = 6) is shown.

Article Snippet: Cells were transfected with a complex consisting of SCP2 siRNA (Santa Cruz Biotechnology, catalog no. sc-44636) and polyamidoamine (PAMAM) G5 (Sigma-Aldrich).

Techniques: Expressing, Staining, Inverted Microscopy, Real-time Polymerase Chain Reaction

Journal: The Journal of Biological Chemistry

Article Title: Sterol carrier protein-2 deficiency attenuates diet-induced dyslipidemia and atherosclerosis in mice

doi: 10.1074/jbc.RA118.002290

Figure Lengend Snippet: Effects of SCP2 deficiency on cholesterol accumulation in and cholesterol efflux from macrophages. Thioglycolate-elicited macrophages were isolated from C57BL/6 (WT) and SCP2−/− mice and incubated with AcLDL (25 μg/ml) for 48 h. A and B, following two washes with PBS, cells were either fixed and stained with Oil Red O (A) or used for total lipid extraction and cholesterol mass measurement (B). Total cholesterol mass was normalized to total cellular protein, and data (mean ± S.D., n = 6) are presented as nanomoles per milligram of protein. C, total protein extracts of macrophages were subjected to Western blot analyses to assess SR-A expression; β-actin was used as a loading control. D, for measurement of cholesterol efflux, cells were loaded with AcLDL and labeled with [3H]-cholesterol for 48 h. Following a 24-h equilibration, cholesterol efflux to 10% FBS in the growth medium was monitored over time. Data (mean ± S.D., n = 6) are presented as percent efflux. *, p < 0.05.

Article Snippet: Cells were transfected with a complex consisting of SCP2 siRNA (Santa Cruz Biotechnology, catalog no. sc-44636) and polyamidoamine (PAMAM) G5 (Sigma-Aldrich).

Techniques: Isolation, Incubation, Staining, Extraction, Mass Measurement, Western Blot, Expressing, Control, Labeling

Journal: The Journal of Biological Chemistry

Article Title: Sterol carrier protein-2 deficiency attenuates diet-induced dyslipidemia and atherosclerosis in mice

doi: 10.1074/jbc.RA118.002290

Figure Lengend Snippet: Effects of SCP2 deficiency on biliary bile acid and cholesterol secretion. A and B, gall bladder bile was collected at the time of euthanasia, and the total volume was noted. Biliary bile acids (BA), cholesterol, and phospholipids (PL) were estimated as described under “Experimental procedures.” Data are presented as total bile acids (nanomoles) or FC (micrograms) in the bile normalized to total phospholipids (micrograms) in A and B, respectively.

Article Snippet: Cells were transfected with a complex consisting of SCP2 siRNA (Santa Cruz Biotechnology, catalog no. sc-44636) and polyamidoamine (PAMAM) G5 (Sigma-Aldrich).

Techniques: