Journal: Journal of the American Chemical Society

Article Title: Encapsulation of Gold-Based Anticancer Agents in Protease-Degradable Peptide Nanofilaments Enhances Their Potency

doi: 10.1021/jacs.2c09820

Figure Lengend Snippet: Cell Viability of gold anticancer agents, gold-loaded peptide nanofilaments ( PD+1 , PD+2 , AD+1 , AD+2 ), and peptides ( PD , AD ) in three human cell lines. Caki-1 (renal cancer), MDA-MB-231 (Triple Negative Breast Cancer), and IMR-90 (lung fibroblasts) incubated with drug-loaded peptide for 72 h. A) Compound 1 , B) Compound 2 . (* represents P <0.05, ** represents P <0.01, *** represents P <0.001).

Article Snippet: Human breast adenocarcinoma line MDA-MB-231 and human lung fibroblast line IMR-90 were obtained from the American Type Culture Collection (ATCC; Manassas, Virginia, USA) and cultured with Dulbecco’s modified Eagle’s medium (DMEM; Fisher Scientific) containing 10 % fetal bovine serum, certified, heat inactivated, US origin (FBS; Fisher Scientific), 1 % minimum essential media (MEM) nonessential amino acids (NEAA; Fisher Scientific), and 1 % penicillin–streptomycin (PenStrep; Fisher Scientific).

Techniques: Incubation

Journal: Vaccine

Article Title: Fluorescence-barcoded cell lines stably expressing membrane-anchored influenza neuraminidases.

doi: 10.1016/j.vaccine.2025.127157

Figure Lengend Snippet: Fig. 1. Standard NA mAbs brightly and specifically label K530-NA cell lines. Shown are flow cytometry histograms depicting the binding of recombinant IgG versions of standard NA mAbs to K530 cell lines expressing membrane-anchored NAs. Each row corresponds to a monoclonal cell line stably expressing a single type of NA. Each column corresponds to the binding profile for a single mAb. MAbs were incubated with pooled cell lines comprising Option 1 or Option 2 (Table 1), and the resultant data were concatenated into a single figure.

Article Snippet: PBMCs in RPMI-1640 medium plus 10 % FBS were incubated with irrelevant mouse IgG1 (MG1K; Rockland) to block nonspecific binding and then labeled with fluorochrome-conjugated mAbs.

Techniques: Flow Cytometry, Binding Assay, Recombinant, Expressing, Membrane, Stable Transfection, Incubation

Journal: Vaccine

Article Title: Fluorescence-barcoded cell lines stably expressing membrane-anchored influenza neuraminidases.

doi: 10.1016/j.vaccine.2025.127157

Figure Lengend Snippet: Fig. 3. Use of K530-NA cells to determine the binding breadth of newly identified NA mAbs. Shown are flow cytometry histograms depicting the binding of re combinant IgG versions of NA mAbs from donors T1, T2, or T3 to K530 cell lines expressing membrane-anchored NAs, as in Fig. 1.

Article Snippet: PBMCs in RPMI-1640 medium plus 10 % FBS were incubated with irrelevant mouse IgG1 (MG1K; Rockland) to block nonspecific binding and then labeled with fluorochrome-conjugated mAbs.

Techniques: Binding Assay, Flow Cytometry, Expressing, Membrane

Journal: Vaccine

Article Title: Fluorescence-barcoded cell lines stably expressing membrane-anchored influenza neuraminidases.

doi: 10.1016/j.vaccine.2025.127157

Figure Lengend Snippet: Fig. 4. Analysis of serum IgGs elicited by infection of rhesus macaques with H3N2 influenza virus. A) Pre-immune (day 0) or immune (day 27 or 28) blood plasma from three rhesus macaques (6451, T651, T771) infected with A/Aichi/2/1968 (H3N2) influenza virus was incubated with K530-NA cell lines. The degree of IgG labeling of each cell line was determined by flow cytometry, as in Fig. 1. Vertical, black lines denote the threshold for specific labeling of cell-surface NA, determined according to the labeling intensity observed for K530 cells expressing no NA (top row). B) Fold-change in the fluorescence intensity of plasma IgG labeling of selected K530-N2 cell lines, after either infection with H3N2 virus (Fig. 4A) or immunization with recombinant H3 HA (Supplementary Fig. 3B). Fold-change was calculated as the ratio of the geometric mean fluorescence intensity (geoMFI) resulting from labeling with the immune plasma IgG, divided by the geoMFI resulting from labeling with pre-immune plasma IgG. Each symbol represents a single animal. The difference in group means was analyzed by two-tailed t-test with Welch’s correction, as described in Materials and Methods.

Article Snippet: PBMCs in RPMI-1640 medium plus 10 % FBS were incubated with irrelevant mouse IgG1 (MG1K; Rockland) to block nonspecific binding and then labeled with fluorochrome-conjugated mAbs.

Techniques: Infection, Virus, Clinical Proteomics, Incubation, Labeling, Flow Cytometry, Expressing, Fluorescence, Recombinant, Two Tailed Test

Journal: Journal of medicinal chemistry

Article Title: 6-Substituted Hexamethylene Amiloride (HMA) Derivatives as Potent and Selective Inhibitors of the Human Urokinase Plasminogen Activator for Use in Cancer

doi: 10.1021/acs.jmedchem.8b00838

Figure Lengend Snippet: Inhibition of low molecular weight human uPA by 18 (black) and 27 (red). Data points represent the mean ± SEM (n = 3) from a single representative experiment.

Article Snippet: Experiments with Mouse uPA (active mouse urokinase, HMW, Molecular Innovations Inc., MI, USA) were performed as above with a final enzyme concentration of 20 nM (K m = 580 μ M).

Techniques: Inhibition, Molecular Weight

Journal: Journal of medicinal chemistry

Article Title: 6-Substituted Hexamethylene Amiloride (HMA) Derivatives as Potent and Selective Inhibitors of the Human Urokinase Plasminogen Activator for Use in Cancer

doi: 10.1021/acs.jmedchem.8b00838

Figure Lengend Snippet: Activities of Compounds 4, 5, 18, and 26 against Human and Mouse uPA

Article Snippet: Experiments with Mouse uPA (active mouse urokinase, HMW, Molecular Innovations Inc., MI, USA) were performed as above with a final enzyme concentration of 20 nM (K m = 580 μ M).

Techniques:

Journal:

Article Title: Identification of a Novel Consensus Sequence at the Cleavage Site of the Lassa Virus Glycoprotein

doi:

Figure Lengend Snippet: Preparation of glycoprotein subunit GP-2 from Lassa virus particles for N-terminal sequencing. (A) Vero-E6 cells were infected with Lassa virus strain Josiah. Virions were purified from the cell culture supernatant by centrifugation through a 20% sucrose cushion followed by iodixanol gradient ultracentrifugation. Fractions of the gradient were SDS treated, subjected to electrophoresis on 12% acrylamide gels, and electrophoretically blotted onto a PVDF membrane. Fractions containing cleaved GP-2 were identified by immunodetection using anti-GP477, horseradish peroxidase-labeled anti-rabbit antibodies from swine (Dako, Glostrup, Denmark), and the Super Signal enhanced chemoluminescence detection kit (Pierce, Rockford, Ill). (B) Proteins of fraction 6 were treated with PNGase F, subjected to SDS-PAGE, and transferred onto a PVDF membrane. Virus glycoproteins GP-1 and GP-2 were detected by immune sera anti-G231 and anti-GP259 (lanes b and c) or were stained with Coomassie blue (lane a). The Coomassie-stained band representing deglycosylated GP-2 was excised and prepared for N-terminal analyses. Molecular mass markers RPN756 used for SDS-PAGE were obtained from Amersham-Pharmacia (Freiburg, Germany).

Article Snippet: Molecular mass markers RPN756 used for SDS-PAGE were obtained from Amersham-Pharmacia (Freiburg, Germany). fig ft0 fig mode=article f1 fig/graphic|fig/alternatives/graphic mode="anchored" m1 Open in a separate window FIG. 3 caption a7 Schematic representation of the Lassa virus glycoprotein and alignment of the amino acid sequences around the cleavage regions of different arenaviruses.

Techniques: Sequencing, Infection, Purification, Cell Culture, Centrifugation, Electrophoresis, Immunodetection, Labeling, SDS Page, Staining

Journal: Neuroscience Bulletin

Article Title: Comprehensive Proteomic Profiling of Patients’ Tears Identifies Potential Biomarkers for the Traumatic Vegetative State

doi: 10.1007/s12264-018-0259-x

Figure Lengend Snippet: Gene Ontology analysis revealed 21 proteins involved in response to wounding.

Article Snippet: ELISA and Receiver Operating Characteristic (ROC) Curve Analysis In the verification stage, the levels of 7 promising tear proteins [cystatin B (CTSB), protease, serine 1 (PRSS1), S100 calcium-binding protein A7 (S100A7), glutathione S-transferase P (GSTP1), complement factor H (CFH), kininogen 1 (KNG1), and alpha-1-acid glycoprotein 1 (ORM1)] were measured using the ELISA kits for human CSTB, human PRSS1, human CFH, human KNG1, and human ORM1 (Boster Biological Technology, Wuhan, China) and for human S100A7 and human GSTP1 (from Cloud-clone Corp., Houston, TX).

Techniques:

Journal: Neuroscience Bulletin

Article Title: Comprehensive Proteomic Profiling of Patients’ Tears Identifies Potential Biomarkers for the Traumatic Vegetative State

doi: 10.1007/s12264-018-0259-x

Figure Lengend Snippet: Levels of representative proteins in tears from healthy controls and traumatic vegetative state patients. A List of 7 selected differentially-expressed proteins. Levels of CTSB (B), PRSS1 (C), S100A7 (D), GSTP1 (E), CFH (F), KNG1 (G), and ORM1 (H).

Article Snippet: ELISA and Receiver Operating Characteristic (ROC) Curve Analysis In the verification stage, the levels of 7 promising tear proteins [cystatin B (CTSB), protease, serine 1 (PRSS1), S100 calcium-binding protein A7 (S100A7), glutathione S-transferase P (GSTP1), complement factor H (CFH), kininogen 1 (KNG1), and alpha-1-acid glycoprotein 1 (ORM1)] were measured using the ELISA kits for human CSTB, human PRSS1, human CFH, human KNG1, and human ORM1 (Boster Biological Technology, Wuhan, China) and for human S100A7 and human GSTP1 (from Cloud-clone Corp., Houston, TX).

Techniques:

Journal: Insect biochemistry and molecular biology

Article Title: Heparan sulfate/heparin glycosaminoglycan binding alters inhibitory profile and enhances anticoagulant function of conserved Amblyomma americanum tick saliva serpin 19

doi: 10.1016/j.ibmb.2016.11.002

Figure Lengend Snippet: List of proteases and their substrates used in substrate hydrolysis assays. Lowest effective protease concentrations (LEPC) and rAAS19 concentrations that inhibit less than 80% of protease activity in specified concentration (IC <80%) are specified.

Article Snippet: The statistical significance of HSGAG binding on rAAS19 inhibitory function was validated using unpaired t-test with Welch's correction in Prism 6 software (GraphPad Software). table ft1 table-wrap mode="anchored" t5 caption a7 Protease (Company) LEPC IC <80% Substrate (Company) Tryptase from Human Lung (Sigma-Aldrich) 4.94 nM NI a N-α-Benzoyl-DL-Arg-pNA (Sigma-Aldrich) Kallikrein from Porcine Pancreas (Sigma-Aldrich) 19.05 nM 1 μM H-D-Pro-Phe-Arg-pNA×2HCl (Chromogenix) Proteinase 3, Human Neutrophil (Athens Research & Technology) 779.31 nM NI a N-Metoxysuccinyl-Ala-Ala-Pro-Val-pNA (Santa Cruz Biotechnology) Elastase from Porcine Pancreas (Sigma-Aldrich) 6.18 nM NI a N-Succinyl-Ala-Ala-Ala-pNA (Sigma-Aldrich) Recombinant Human HMW Urokinase (Molecular Innovations) 9.26 nM NI a CH3SO2-D-CHG-Gly-Arg-pNA×AcOH (Pentapharm) Human tPA, >85% Single Chain (Molecular Innovations) 4.73 nM NI a CH3SO2-D-CHG-Gly-Arg-pNA×AcOH (Pentapharm) Papain (Sigma-Aldrich) 2.14 μM 1 μM N-Benzoyl-Phe-Val-Arg-pNA×HCl (Sigma-Aldrich) Bovine Factor IXa beta (Enzyme Research Laboratories) 31.44 nM 1 μM CH3SO2-D-CHG-Gly-Arg-pNA×AcOH (Pentapharm) Factor Xa Protease (New England Biolabs) 4.65 nM 40 nM Benzoyl-Ile-Glu(γ-OR)-Gly-Arg-pNA×HCl (Chromogenix) Human Factor XIa (Enzyme Research Laboratories) 3.69 nM 200 nM H-D-Pro-Phe-Arg-pNA×2HCl (Chromogenix) Human Factor alpha-XIIa (Enzyme Research Laboratories) 2.03 nM 1 μM H-D-Pro-Phe-Arg-pNA×2HCl (Chromogenix) Plasmin from Human Plasma (Sigma-Aldrich) 11.63 nM 20 nM H-D-Val-Leu-Lys-pNA×2HCl (Chromogenix) Thrombin from Bovine Plasma (Sigma-Aldrich) 27.1 nM 100 nM H-D-Phe-Pip-Arg-pNA×2HCl (Chromogenix) Trypsin from Bovine Pancreas (Sigma-Aldrich) 21.01 nM 4 nM Benzoyl-Ile-Glu(γ-OR)-Gly-Arg-pNA×HCl (Chromogenix) Open in a separate window a No inhibition List of proteases and their substrates used in substrate hydrolysis assays.

Techniques: Activity Assay, Concentration Assay, Recombinant