Journal: International Journal of Molecular Sciences

Article Title: Structural Characterization of Glycoprotein Glycans and Glycosaminoglycans of Brain Tissues in Slc35a3 -Knockout Mice

doi: 10.3390/ijms27041643

Figure Lengend Snippet: Brain N -glycome in wild-type, hetero, and Slc35a3 -KO mice. ( a ) Representative MALDI-TOF mass spectra of PNGase F-released N -glycans from whole brain for wild-type, hetero, and Slc35a3 -KO mice. Individual peak intensity was normalized to the internal standard (I.S.). Annotated peaks are shown as predicted glycan structures, illustrated with SNFG cartoons. ( b ) Classwise quantification of N -glycans (pmol per 100 µg protein): paucimannose type, high-mannose type, hybrid type, neutral complex type, and acidic complex type. ( c ) Branching distribution of N -glycans expressed as the number of HexNAc residues added beyond the chitobiose core (Core + HexNAc0–4). Bars: mean ± SD; colors denote genotype (wild type, white; hetero, blue; KO, red), (ns, not significant; *, p < 0.05; **, p < 0.01 and ***, p < 0.001; unpaired two-tailed Student’s t test). Individual data points are shown as black dots overlaid on the bar graph. Representative glycan structures for each class are illustrated as SNFG cartoons above the graph.

Article Snippet: To verify N -glycan dependence of lectin binding, lysates were treated with PNGase F (500 U per 20 μg protein; New England Biolabs, Ipswich, MA, USA) to remove N -glycans, or with Endoglycosidase H (Endo H; 500 U per 20 μg protein; Seikagaku Kogyo Co., Ltd., Tokyo, Japan) to selectively cleave high-mannose and hybrid-type N -glycans, prior to electrophoresis.

Techniques: Glycoproteomics, Two Tailed Test

Journal: International Journal of Molecular Sciences

Article Title: Structural Characterization of Glycoprotein Glycans and Glycosaminoglycans of Brain Tissues in Slc35a3 -Knockout Mice

doi: 10.3390/ijms27041643

Figure Lengend Snippet: Lectin blotting analysis for brain tissues from wild-type (WT) and Slc35a3 -KO (KO) mice using ( a , b ) concanavalin A (ConA) and ( c , d ) peanut agglutinin (PNA). ( a ) Homogenized brain lysates from wild-type and Slc35a3 -KO mice were enzymatically digested with PNGase F or Endo H and subjected to lectin blotting using ConA, which recognizes high-mannose-type N -glycans. ( c ) Brain lysates prepared as in ( a ) were enzymatically digested with O -glycosidase and/or neuraminidase and analyzed by lectin blotting using PNA, which recognizes Galβ1–3GalNAc structures. β-Actin was used as a loading control to verify equal protein loading. ( b , d ) Quantification of ConA ( b ) and PNA ( d ) signals shown in the lectin blots in ( a , c ) and . The raw data are provided in . Lectin blot intensities above 48 kDa were quantified for the entire lane and normalized to β-actin. Signal intensities are expressed relative to the WT control. Data represent three independent experiments ( n = 3); bars indicate mean ± SD. Statistical significance was assessed as indicated (ns, not significant; **, p < 0.01 and ***, p < 0.001; two-way ANOVA). Individual data points are plotted as black dots on the bar graph.

Article Snippet: To verify N -glycan dependence of lectin binding, lysates were treated with PNGase F (500 U per 20 μg protein; New England Biolabs, Ipswich, MA, USA) to remove N -glycans, or with Endoglycosidase H (Endo H; 500 U per 20 μg protein; Seikagaku Kogyo Co., Ltd., Tokyo, Japan) to selectively cleave high-mannose and hybrid-type N -glycans, prior to electrophoresis.

Techniques: Control

Journal: Antimicrobial Agents and Chemotherapy

Article Title: False-Positive Carbapenem-Hydrolyzing Confirmatory Tests Due to ACT-28, a Chromosomally Encoded AmpC with Weak Carbapenemase Activity from Enterobacter kobei

doi: 10.1128/AAC.02388-18

Figure Lengend Snippet: Results of carbapenemase detection testsa

Article Snippet: Among 1,039 non-carbapenemase-producing ECC isolates with decreased susceptibility to carbapenems received in 2016-2017 at the French National Reference Center for antibiotic resistance, only 8 had a positive carbapenemase detection test (Carba NP).

Techniques:

Journal: Antimicrobial Agents and Chemotherapy

Article Title: False-Positive Carbapenem-Hydrolyzing Confirmatory Tests Due to ACT-28, a Chromosomally Encoded AmpC with Weak Carbapenemase Activity from Enterobacter kobei

doi: 10.1128/AAC.02388-18

Figure Lengend Snippet: Phylogenetic analysis of AmpC β-lactamases of ECC isolates addressed to the French National Reference Center for carbapenemase detection. This unrooted tree was constructed based on ampC genes using the maximum likelihood method with the Tamura-Nei model using the MEGA program (MEGA 7.0) The tree is drawn to scale, with branch lengths representing the evolutionary distances. ACT-28-producing ECC isolates from the NCBI database (1323, e1425, MGH25, MGH37, ICBEaBL-111-03-02, CRE54, CRE71, and GN02825) and ampC of ECC genomes of Chavda’s phylogenomic groups B, C, E, J, G, H, and Q were added to the analysis (23). NR, not realized.

Article Snippet: Among 1,039 non-carbapenemase-producing ECC isolates with decreased susceptibility to carbapenems received in 2016-2017 at the French National Reference Center for antibiotic resistance, only 8 had a positive carbapenemase detection test (Carba NP).

Techniques: Construct

Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

Article Title: Gas Chromatography-Ion Mobility Spectrometry Reveals Acetoin as a Biomarker for Carbapenemase-Producing Klebsiella pneumoniae

doi: 10.12659/MSM.944507

Figure Lengend Snippet: The changes in the content of 3-hydroxy-2-butanone (acetoin) emitted by standard strain at the T2 time point before and after the addition of carbapenemase inhibitors. ( A ) The pseudo-3D plots of 3-hydroxy-2-butanone (acetoin) contents in standard strains before and after the addition of carbapenase inhibitors. VOCal (version 0.1.3), G.A.S., Shandong HaiNeng Scientific Instrument Co., Ltd., Shandong, China. ( B ) Comparison of the relative peak volume (semiquantitative) of 3-hydroxy-2-butanone (acetoin) emitted by standard strains before and after the addition of carbapenase inhibitors. The free online tool Chiplot ( https://www.chiplot.online ). The Mann-Whitney U test was used for pairwise comparison. * P <0.05. R version 4.2.3, The R Foundation, Vienna, Austria. ATCC – American Type Culture Collection; IPM – imipenem; DPA – pyridine-2,6-dicarboxylic acid.

Article Snippet: Currently, the laboratory tests for carbapenemases rely on the modified carbapenem inactivation method (mCIM) and the Carbapenemase Non-Phenotypic (Carba NP) test, as recommended by the Clinical and Laboratory Standards Institute (CLSI) [ , ].

Techniques: Comparison, MANN-WHITNEY