Journal: Cells

Article Title: A Neutrophil-like Cell Model as Substitute for Human Neutrophils in NETs and Thrombosis Research

doi: 10.3390/cells15060541

Figure Lengend Snippet: Structural analysis of differentiated PLB-985 and human neutrophil cells. SEM images of normal PLB-985 cells ( a , b ), a differentiated PLB-985 cell ( c ), and a human neutrophil ( d ). Scale bars are 5 µm (n ≥ 3). SEM analysis of cell diameter (( e ), n ≥ 21) and cell surface filament diameter (( f ), n ≥ 83). Error bars represent mean ± SD. ** p < 0.01, **** p < 0.0001, ns: non-significant. ( e , f ) The statistical analysis results were obtained using the non-parametric Kruskal–Wallis test followed by Dunn’s multiple comparisons test. Experiments were performed independently at least three times using separate cell preparations.

Article Snippet: The PLB-985 cell line (ACC-139, DSMZ, Braunschweig, Germany) was cultured and differentiated, using medium containing 1.25% DMSO, as described previously [ ].

Techniques:

Journal: Cells

Article Title: A Neutrophil-like Cell Model as Substitute for Human Neutrophils in NETs and Thrombosis Research

doi: 10.3390/cells15060541

Figure Lengend Snippet: Analysis of surface morphology and mechanical properties of differentiated PLB-985 cells and their NETs. Air-dried differentiated PLB-985 cells ( a – c ) and air-dried PLB-985 NETs ( d – h ). ( a ) The amplitude error channel reflects a three-dimensional image of cells. ( b ) The height channel reflects the size range of cells. ( c ) The height profile taken along the blue line in panel. ( d ) The amplitude error channel reflects a three-dimensional image of NETs. ( e ) The height channel reflects the size range of NETs. ( f ) The height profile taken along the blue line in panel ( d , e ). ( g ) The height profile taken along the white line in panel ( d , e ). ( h ) The height profile taken along the green line in panel ( d , e ). Experiments were performed independently at least three times using separate cell preparations.

Article Snippet: The PLB-985 cell line (ACC-139, DSMZ, Braunschweig, Germany) was cultured and differentiated, using medium containing 1.25% DMSO, as described previously [ ].

Techniques:

Journal: Cells

Article Title: A Neutrophil-like Cell Model as Substitute for Human Neutrophils in NETs and Thrombosis Research

doi: 10.3390/cells15060541

Figure Lengend Snippet: Structural analysis of differentiated PLB-985 and human neutrophil NETs. SEM images of PLB-985 NETs ( a ) and human NETs ( b ). Scale bars are 2 μm. Red circle: spherical particles, either individually or in a series (n ≥ 3). SEM analysis of NET fibre diameter (( c ), n ≥ 514) and the diameter of spherical particles in PLB samples (( d ), n = 261). Error bars represent mean ± SD. **** p < 0.0001. ( c ) The statistical analysis results were obtained using the non-parametric Mann–Whitney test. Experiments were performed independently at least three times using separate cell preparations.

Article Snippet: The PLB-985 cell line (ACC-139, DSMZ, Braunschweig, Germany) was cultured and differentiated, using medium containing 1.25% DMSO, as described previously [ ].

Techniques: MANN-WHITNEY

Journal: Cells

Article Title: A Neutrophil-like Cell Model as Substitute for Human Neutrophils in NETs and Thrombosis Research

doi: 10.3390/cells15060541

Figure Lengend Snippet: Structural analysis of clots formed with differentiated PLB-985 and human neutrophil NETs. SEM images of PLB-985 NET-induced and human NET-induced plasma clots ( a ). Thrombin-induced clots were used as a control. Final concentrations: plasma (diluted 1:3), 10 mM CaCl 2 , and 1 U/mL thrombin. Scale bars are 5 µm. Red circle: spherical particles (n ≥ 3). Effects of human NETs and PLB-985 NETs on the fibrin fibre thickness in plasma ( b ). Final concentrations: plasma (diluted 1:6), cells (200,000 cells/100 µL), NETs (generated from 200,000 cells/100 µL), thrombin (0.1 U/mL), and CaCl 2 (3.33 mM). Error bars represent mean ± SD (n = 80). **** p < 0.0001, using the non-parametric Kruskal–Wallis test followed by Dunn’s multiple comparisons test. Experiments were performed independently at least three times using separate cell preparations.

Article Snippet: The PLB-985 cell line (ACC-139, DSMZ, Braunschweig, Germany) was cultured and differentiated, using medium containing 1.25% DMSO, as described previously [ ].

Techniques: Clinical Proteomics, Control, Generated

Journal: Life

Article Title: Almond Skin Extracts and Chlorogenic Acid Delay Chronological Aging and Enhanced Oxidative Stress Response in Yeast

doi: 10.3390/life10060080

Figure Lengend Snippet: Effects of almond skin extract (AE) and CHL on SIR2 ( a ), SOD1 ( b ) and SOD2 ( c ) gene expression determined by RT-qPCR. Expression was normalized with TUB1 gene and expressed relative to young yeast cells (Y). Y CTL was young yeast (on day 2 of cultivation) and A CTL was aged yeast (on day 5 of cultivation). AE, almond extract (1 mg/mL). CHL: chlorogenic acid at 3 concentrations (CHL5, CHL10 and CHL25 corresponding to chlorogenic acid addition at 5, 10 and 25 µM, respectively). E-Resveratrol (RES, 10 µM) was used as control antiaging drug. Values are means ± standard deviations (SD) of 4 independent experiments. Different letters represent significant differences between the various extraction conditions ( p < 0.05).

Article Snippet: Chlorogenic acid (CHL) and resveratrol (RES) standard was purchased from LGC Standard (Molsheim, France).

Techniques: Gene Expression, Quantitative RT-PCR, Expressing, Control, Extraction

Journal: Life

Article Title: Almond Skin Extracts and Chlorogenic Acid Delay Chronological Aging and Enhanced Oxidative Stress Response in Yeast

doi: 10.3390/life10060080

Figure Lengend Snippet: Effects of AE and CHL on SIRT1/SIR2 ( a ) and total SOD ( b ) enzyme activities. Y-CTL was young yeast (on day 2 of cultivation) and A-CTL was aged yeast (on day 5 of cultivation). AE, almond extract (1 mg/mL). CHL: chlorogenic acid at 3 concentrations (CHL5, CHL10 and CHL25 corresponding to chlorogenic acid addition at 5, 10 and 25 µM, respectively). E-Resveratrol (RES, 10 µM) was used as control antiaging drug. In young yeast culture (Y), actual SIRT1/SIR2 enzyme activity was equal to 4564.4 FU/mg protein, whereas total SOD activity was 35.6 units/mg protein. The enzyme activity was expressed as a percentage of Y-CTL. Values are means ± standard deviations (SD) of 4 independent experiments. Different letters represent significant differences between the various extraction conditions ( p < 0.05).

Article Snippet: Chlorogenic acid (CHL) and resveratrol (RES) standard was purchased from LGC Standard (Molsheim, France).

Techniques: Control, Activity Assay, Extraction

Journal: Life

Article Title: Almond Skin Extracts and Chlorogenic Acid Delay Chronological Aging and Enhanced Oxidative Stress Response in Yeast

doi: 10.3390/life10060080

Figure Lengend Snippet: Survival and ROS/RNS production under UV-C-induced oxidative stress conditions. ( a ) Survival assay, plates were incubated at 28 °C for 3 d and the number of colonies was counted; excepted if mentioned CTL was DBY746 yeast subjected to UV-C stress. ( b ) ROS/RNS production was evaluated using the dihydrorhodamine 123 (DHR123) probe. AE, almond extract (1 mg/mL). CHL: chlorogenic acid at 3 concentrations (CHL5, CHL10 and CHL25 corresponding to chlorogenic acid addition at 5, 10 and 25 µM, respectively). E-Resveratrol (RES, 10 µM) was used as control antiaging drug. Values are means ± standard deviations (SD) of 6 independent experiments. Different letters represent significant differences between the various extraction conditions ( p < 0.05).

Article Snippet: Chlorogenic acid (CHL) and resveratrol (RES) standard was purchased from LGC Standard (Molsheim, France).

Techniques: Clonogenic Cell Survival Assay, Incubation, Control, Extraction

Journal: Life

Article Title: Almond Skin Extracts and Chlorogenic Acid Delay Chronological Aging and Enhanced Oxidative Stress Response in Yeast

doi: 10.3390/life10060080

Figure Lengend Snippet: Schematic view of the different ROS/RNS-induced oxidative damages to membrane lipids, proteins and DNA ( a ), and effects of AE and CHL on malondialdehyde (MDA) levels measured by TBARS assay ( b ), protein carbonyl contents determined by ELISA assay ( c ) and 8-oxo-Guanine formation determined by ELISA assay ( d ). AE, almond extract (1 mg/mL). CHL: chlorogenic acid at 3 concentrations (CHL5, CHL10 and CHL25 corresponding to chlorogenic acid addition at 5, 10 and 25 µM, respectively). E-Resveratrol (RES, 10 µM) was used as control antiaging drug. Values are means ± standard deviations (SD) of 6 independent experiments. Different letters represent significant differences between the various extraction conditions ( p < 0.05).

Article Snippet: Chlorogenic acid (CHL) and resveratrol (RES) standard was purchased from LGC Standard (Molsheim, France).

Techniques: Membrane, TBARS Assay, Enzyme-linked Immunosorbent Assay, Control, Extraction