Journal: Journal of Clinical Microbiology

Article Title: Comparison of the Vitek MS and Bruker Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry Systems for Identification of Chryseobacterium Isolates from Clinical Specimens and Report of Uncommon Chryseobacterium Infections in Humans

doi: 10.1128/JCM.00712-18

Figure Lengend Snippet: Chryseobacterium identified at the species level by the Vitek MS and Bruker Biotyper systems compared to the results of 16S rRNA gene sequencing

Article Snippet: Among the 129 isolates identified at the species level, 78 and 39 isolates were identified as C. indologenes (type strain DSM 16777 T ; GenBank accession no. {"type":"entrez-nucleotide","attrs":{"text":"LN681561","term_id":"732170053","term_text":"LN681561"}} LN681561 ) and C. gleum (type strain ATCC 35910 T ; GenBank accession no. {"type":"entrez-nucleotide","attrs":{"text":"GL379781","term_id":"301087312","term_text":"GL379781"}} GL379781 ), respectively ( ).

Techniques: Sequencing

Journal: Journal of Clinical Microbiology

Article Title: Comparison of the Vitek MS and Bruker Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry Systems for Identification of Chryseobacterium Isolates from Clinical Specimens and Report of Uncommon Chryseobacterium Infections in Humans

doi: 10.1128/JCM.00712-18

Figure Lengend Snippet: Chryseobacterium species misidentified by the Vitek MS and Bruker Biotyper systems compared to the results of 16S rRNA gene sequencing

Article Snippet: Among the 129 isolates identified at the species level, 78 and 39 isolates were identified as C. indologenes (type strain DSM 16777 T ; GenBank accession no. {"type":"entrez-nucleotide","attrs":{"text":"LN681561","term_id":"732170053","term_text":"LN681561"}} LN681561 ) and C. gleum (type strain ATCC 35910 T ; GenBank accession no. {"type":"entrez-nucleotide","attrs":{"text":"GL379781","term_id":"301087312","term_text":"GL379781"}} GL379781 ), respectively ( ).

Techniques: Sequencing

Journal: Sensors (Basel, Switzerland)

Article Title: Combination of Aptamer Amplifier and Antigen-Binding Fragment Probe as a Novel Strategy to Improve Detection Limit of Silicon Nanowire Field-Effect Transistor Immunosensors

doi: 10.3390/s21020650

Figure Lengend Snippet: Illustration of the detection of rabbit IgG by (1) Wab/APTES-SiNWFETs, (2) Fab/APTES-SiNWFETs, (3) Wab/PEG-SiNWFETs, and (4) Fab/PEG-SiNWFETs as well as their corresponding signal enhancement by R18 aptamer in this study. Both of the Wab/APTES-SiNWFETs (sample 1) and Fab/APTES-SiNWFETs (sample 2) were used to detect ( A ) rabbit IgG (blue Y shapes) at concentrations of 100 pg/mL and 1 ng/mL, before binding with ( B ) 3 μg/mL R18 aptamer (green curves) for signal enhancement. The Wab/PEG-SiNWFETs (sample 3) could only determine ( A ) rabbit IgG (blue Y shapes) at concentrations of 10 pg/mL, 100 pg/mL and 1 ng/mL, whereas the Fab/PEG-SiNWFETs (sample 4) could recognize ( A ) rabbit IgG (blue Y shapes) at concentrations of 1 pg/mL, 10 pg/mL, 100 pg/mL, and 1 ng/mL. Both of them were then also incubated in ( B ) 3 μg/mL R18 aptamer (green curves) for signal enhancement. All the biosensing experiments in this Figure were performed in 150 mM BTP.

Article Snippet: 3,3′,5,5′-Tetramethylbenzidine (TMB), acetone, and ethanol (99.9%) was ordered from Thermo Fisher Scientific, while the components of PEG-mSAMs (silane-PEG-NH 2 , 1K and silane-PEG-OH, 1K) were from Biochempeg Scientific Inc. All the antibodies, including rabbit immunoglobulin G (IgG), donkey anti-goat antibody conjugated with horseradish peroxide (HRP), goat anti-rabbit IgG antibody (Wab), and its Fab, were supplied by Abcam plc. (UK), whereas R18 aptamer specifying rabbit IgG (74-mer RNA sequence: 5′-GGGAG AAUUC CGACC AGAAG UUCGA UACGC CGUGG GGUGA CGUUG GCUAC CCUUU CCUCU CUCCU CCUUC UUCU-3′ [ ]) was synthesized by MDBio, Inc.

Techniques: Binding Assay, Incubation

Journal: Sensors (Basel, Switzerland)

Article Title: Combination of Aptamer Amplifier and Antigen-Binding Fragment Probe as a Novel Strategy to Improve Detection Limit of Silicon Nanowire Field-Effect Transistor Immunosensors

doi: 10.3390/s21020650

Figure Lengend Snippet: ( A , B ) Representative samples to illustrate the method described in . ( A ) Electrical response of the Wab/APTES-SiNWFETs was initially recorded in 150 mM BTP and plotted as the first curve (the black curve). This immunosensor was then employed to detect rabbit IgG at 1 ng/mL, followed by incubation with 3 μg/mL R18 (IgG and R18 were diluted in 150 mM BTP). Finally, its electrical response was measured again and plotted as the blue curve. The signal change generated by formation of the biocomplex (Wab-IgG-R18) was calculated from the formula ΔV = V d1 − V d0 (1), with V d1 as the gate voltage value at I d = 10 −9 A (LgI = −9) of the blue curve, whereas V d0 is the gate voltage value at I d = 10 −9 A (LgI = −9) of the black curve. ( B ) Electrical response of the Fab/APTES-SiNWFET was initially recorded in 150 mM BTP and plotted as the first curve (the black curve). This immunosensor was then employed to detect rabbit IgG at 1 ng/mL following by incubation with 3 μg/mL R18 (IgG and R18 were all diluted in 150 mM BTP). Finally, its electrical response was measured again and plotted as the blue curve. The signal change generated by formation of the biocomplex (Fab-IgG-R18) was calculated from the formula ΔV = V d1 − V d0 (1), with V d1 is the gate voltage value at I d = 10 −9 A (LgI = −9) of the blue curve, whereas V d0 is the gate voltage value at I d = 10 −9 A (LgI = −9) of the black curve. ( C ) Comparison of the signal amplified by R18 (mV) after determining rabbit IgG at different concentrations (0.1 ng/mL and 1 ng/mL) in 150 mM BTP by Wab/APTES-SiNWFETs (blue bars) and Fab/APTES-SiNWFETs (red bars). The voltage shift (mV) generated by IgG detection of APTES-SiNWFETs without probes (Wab nor Fab) (black bar), and by recognizing R18 without IgG (0 ng/mL) of Fab/APTES-SiNWFETs, was also calculated for analysis.

Article Snippet: 3,3′,5,5′-Tetramethylbenzidine (TMB), acetone, and ethanol (99.9%) was ordered from Thermo Fisher Scientific, while the components of PEG-mSAMs (silane-PEG-NH 2 , 1K and silane-PEG-OH, 1K) were from Biochempeg Scientific Inc. All the antibodies, including rabbit immunoglobulin G (IgG), donkey anti-goat antibody conjugated with horseradish peroxide (HRP), goat anti-rabbit IgG antibody (Wab), and its Fab, were supplied by Abcam plc. (UK), whereas R18 aptamer specifying rabbit IgG (74-mer RNA sequence: 5′-GGGAG AAUUC CGACC AGAAG UUCGA UACGC CGUGG GGUGA CGUUG GCUAC CCUUU CCUCU CUCCU CCUUC UUCU-3′ [ ]) was synthesized by MDBio, Inc.

Techniques: Incubation, Generated, Comparison, Amplification

Journal: Sensors (Basel, Switzerland)

Article Title: Combination of Aptamer Amplifier and Antigen-Binding Fragment Probe as a Novel Strategy to Improve Detection Limit of Silicon Nanowire Field-Effect Transistor Immunosensors

doi: 10.3390/s21020650

Figure Lengend Snippet: ( A ) Comparison of the signal amplified by R18 (mV) after sensing rabbit IgG at various levels in 150 mM BTP by Wab/PEG-SiNWFETs (blue bars) and Fab/APTES-SiNWFETs (red bars). ( B ) Plot of the voltage shift by R18 versus logarithmic concentrations of rabbit IgG and two respective calibration lines obtained by Wab/PEG-SiNWFETs (blue line) and Fab/APTES-SiNWFETs (red line).

Article Snippet: 3,3′,5,5′-Tetramethylbenzidine (TMB), acetone, and ethanol (99.9%) was ordered from Thermo Fisher Scientific, while the components of PEG-mSAMs (silane-PEG-NH 2 , 1K and silane-PEG-OH, 1K) were from Biochempeg Scientific Inc. All the antibodies, including rabbit immunoglobulin G (IgG), donkey anti-goat antibody conjugated with horseradish peroxide (HRP), goat anti-rabbit IgG antibody (Wab), and its Fab, were supplied by Abcam plc. (UK), whereas R18 aptamer specifying rabbit IgG (74-mer RNA sequence: 5′-GGGAG AAUUC CGACC AGAAG UUCGA UACGC CGUGG GGUGA CGUUG GCUAC CCUUU CCUCU CUCCU CCUUC UUCU-3′ [ ]) was synthesized by MDBio, Inc.

Techniques: Comparison, Amplification

Journal: Leukemia

Article Title: IDH1 and IDH2 mutations in pediatric acute leukemia

doi: 10.1038/leu.2011.133

Figure Lengend Snippet: Genetic characteristics of the 515 pediatric leukemias analyzed

Article Snippet: 20 Site Directed Mutagenesis Human wildtype (WT) cDNA clones for IDH1 ( {"type":"entrez-nucleotide","attrs":{"text":"NM_005896.2","term_id":"28178824","term_text":"NM_005896.2"}} NM_005896.2 ) and IDH2 ( {"type":"entrez-nucleotide","attrs":{"text":"NM_002168.2","term_id":"28178831","term_text":"NM_002168.2"}} NM_002168.2 ) in pCMV6 Entry vector were obtained from Origene (Origene, Rockville, MD).

Techniques:

Journal: Leukemia

Article Title: IDH1 and IDH2 mutations in pediatric acute leukemia

doi: 10.1038/leu.2011.133

Figure Lengend Snippet: Genomic structure of IDH1 and IDH2 and structural model of the location of R140 and R172 in the substrate-binding pocket of IDH2. (a) Illustration of the exon/intron structure of IDH1 and IDH2. The location within exon 4 of codon R132 in IDH1 and codon R140 in IDH2 are marked by arrows, and the surrounding nucleotide sequence and encoded amino acids are highlighted. Codon R132 in IDH1 and the homologous residue R172 in IDH2 are shown in red and codon 140 in IDH2 is shown in blue. (b) Model of the positions of the R140 and R172 amino acids in the IDH2 substrate binding pocket. The two protomers in the IDH2 homodimer are illustrated in green and purple, with the nitrogen atoms in amino acids R140 and R172 shown in blue. The bound substrate, isocitrate, is depicted with carbon atoms as yellow sticks and oxygen atoms in red. The manganese ion is shown as a grey sphere. Salt-bridges and hydrogen bonds are shown as dashed lines.

Article Snippet: 20 Site Directed Mutagenesis Human wildtype (WT) cDNA clones for IDH1 ( {"type":"entrez-nucleotide","attrs":{"text":"NM_005896.2","term_id":"28178824","term_text":"NM_005896.2"}} NM_005896.2 ) and IDH2 ( {"type":"entrez-nucleotide","attrs":{"text":"NM_002168.2","term_id":"28178831","term_text":"NM_002168.2"}} NM_002168.2 ) in pCMV6 Entry vector were obtained from Origene (Origene, Rockville, MD).

Techniques: Binding Assay, Sequencing

Journal: Leukemia

Article Title: IDH1 and IDH2 mutations in pediatric acute leukemia

doi: 10.1038/leu.2011.133

Figure Lengend Snippet: Genetic characteristics of the AMLs with IDH1 / IDH2 mutations

Article Snippet: 20 Site Directed Mutagenesis Human wildtype (WT) cDNA clones for IDH1 ( {"type":"entrez-nucleotide","attrs":{"text":"NM_005896.2","term_id":"28178824","term_text":"NM_005896.2"}} NM_005896.2 ) and IDH2 ( {"type":"entrez-nucleotide","attrs":{"text":"NM_002168.2","term_id":"28178831","term_text":"NM_002168.2"}} NM_002168.2 ) in pCMV6 Entry vector were obtained from Origene (Origene, Rockville, MD).

Techniques: Mutagenesis

Journal: Leukemia

Article Title: IDH1 and IDH2 mutations in pediatric acute leukemia

doi: 10.1038/leu.2011.133

Figure Lengend Snippet: Enzymatic analysis of the IDH1 and IDH2 mutant proteins. The activity of recombinant IDH1 and IDH2 proteins to catalyze the NADP+-dependent oxidative decarboxylation of isocitrate to α-KG using 30uM Isocitrate and 100uM NADP are shown in panels a and b, respectively, and their ability to catalyze the NADPH-dependent reduction of α–KG to 2-HG using 0.5 mM α-KG and 100uM NADPH are shown in c and d, respectively. All graphs are based on triplicate measurements with the mean ± standard deviations expressed as a relative level compared to WT:WT homodimers, with the latter set as 100%. The SD in panel c and d are < 0.03 and are thus below the resolution of the figure. (e) The intracellular level of 2-HG was measured by liquid chromatography/mass spectrometry in pediatric AML cells from primary diagnostic bone marrow samples. The data for mutant IDH1/IDH2 includes two leukemia samples containing IDH1 mutations (one with R132H and one with R132C, ▴), and two containing the R140Q IDH2 mutations (◆). The wild-type IDH1/IDH2 data was generated using two pediatric AML samples that lacked mutations in either IDH1 or IDH2 (●).

Article Snippet: 20 Site Directed Mutagenesis Human wildtype (WT) cDNA clones for IDH1 ( {"type":"entrez-nucleotide","attrs":{"text":"NM_005896.2","term_id":"28178824","term_text":"NM_005896.2"}} NM_005896.2 ) and IDH2 ( {"type":"entrez-nucleotide","attrs":{"text":"NM_002168.2","term_id":"28178831","term_text":"NM_002168.2"}} NM_002168.2 ) in pCMV6 Entry vector were obtained from Origene (Origene, Rockville, MD).

Techniques: Mutagenesis, Activity Assay, Recombinant, Liquid Chromatography, Mass Spectrometry, Diagnostic Assay, Generated