Journal: Scientific Reports

Article Title: Dysfunction of low-density neutrophils in peripheral circulation in patients with sepsis

doi: 10.1038/s41598-021-04682-x

Figure Lengend Snippet: Reduced chemotactic function of LDNs. ( A ) Neutrophil chemotaxis toward fMLP was assayed. CD (chemotactic distance, μm), CCR (chemo cell ratio, cell number of zone I + II + III/Total cells in the side well, %), CI (chemo index, cell number of zone II + III/ Total number of chemotactic cells, %). ( B – D ) The evaluation indicators CD, CCR, and CI were significantly reduced in neutrophil chemotactic function evaluations of LDN. ( E ) The expression of P2X1 on HDNs and LDNs was analyzed using flow cytometry.

Article Snippet: Antibodies used in the experiment include CD66b (G10F5, BD Pharmingen, USA), CD10 (HI10α, Biolegend, USA) CXCR4 (12G5, Biolegend, USA) and P2X1 (APR-022-F, Alomone labs, Israel).

Techniques: Chemotaxis Assay, Expressing, Flow Cytometry

Journal: Critical Care

Article Title: Erythrocyte P2X 1 receptor expression is correlated with change in haematocrit in patients admitted to the ICU with blood pathogen-positive sepsis

doi: 10.1186/s13054-018-2100-3

Figure Lengend Snippet: Model of pore former induced lysis. A bacterial toxin inserts a large channel or pore into the erythrocyte membrane. ATP is immediately released through the pore and activates P2X receptors. The membrane insertion of the toxin also causes a steep rise in the intracellular Ca 2+ concentration ([Ca 2+ ] i ), which results from Ca 2+ passing through the pore itself and from activation of P2X receptors, which are non-selective cation channels permeable to Ca 2+ . The increase in [Ca 2+ ] i activates the Ca 2+ -sensitive K + channel K Ca 3.1 and Cl − channel TMEM16A, which results in K + and Cl − efflux and cell shrinkage as obligated water follows. The cells will remain shrunken as long as the K + efflux surpasses the Na + influx via the toxin pore and the P2X receptors. Prolonged stimulation of P2X 7 can activate pannexins, which will contribute to the Na + influx. Eventually, the Na + influx will exceed the K + efflux and the cells will swell and eventually burst. Blockage of the P2X 1 and P2X 7 receptor has been proven as a protective measure for bacterial toxins, and for complement to carry out their toxicity. The model based on previous work [ , , , ]

Article Snippet: Cells in each sample were counted on a cell counter (Spectre, Merch Millipore, USA) prior to exposure to either P2X 1 (catalogue number APR-022-AG) or P2X 7 (catalogue number APR-008-F) fluorescein isothiocyanate (FITC)-conjugated antibody (Alomone Labs, Jerusalem, Israel) for 1 h at room temperature in the dark at 200 rpm in concentrations according to the manufacturer’s instructions (10 mg antibody to 10 6 cells).

Techniques: Lysis, Concentration Assay, Activation Assay

Journal: Critical Care

Article Title: Erythrocyte P2X 1 receptor expression is correlated with change in haematocrit in patients admitted to the ICU with blood pathogen-positive sepsis

doi: 10.1186/s13054-018-2100-3

Figure Lengend Snippet: Bacterial origin

Article Snippet: Cells in each sample were counted on a cell counter (Spectre, Merch Millipore, USA) prior to exposure to either P2X 1 (catalogue number APR-022-AG) or P2X 7 (catalogue number APR-008-F) fluorescein isothiocyanate (FITC)-conjugated antibody (Alomone Labs, Jerusalem, Israel) for 1 h at room temperature in the dark at 200 rpm in concentrations according to the manufacturer’s instructions (10 mg antibody to 10 6 cells).

Techniques: In Vitro

Journal: Critical Care

Article Title: Erythrocyte P2X 1 receptor expression is correlated with change in haematocrit in patients admitted to the ICU with blood pathogen-positive sepsis

doi: 10.1186/s13054-018-2100-3

Figure Lengend Snippet: Freeze-thaw - control experiments. a Whole blood drawn from healthy volunteers was incubated either with vehicle (Veh), 100 μM NF449 (P2X 1 antagonist) or 100 μM A804598 (P2X 7 antagonist) and frozen and stored for 3 weeks as whole blood at − 80 °C. After thawing, lysis was measured as absorbance of free haemoglobin in supernatant at 540 nm. n = 3, mean ± SEM. b Flow cytometry of thawed whole blood samples diluted 1000-fold, to ascertain equal distribution of blood cell sub-populations. Cells were identified by forward scatter FSC and side scatter SSC and quantified based on fixed regions. c Bar graph shows data presented as mean ± SEM, n = 3. ns, not significant

Article Snippet: Cells in each sample were counted on a cell counter (Spectre, Merch Millipore, USA) prior to exposure to either P2X 1 (catalogue number APR-022-AG) or P2X 7 (catalogue number APR-008-F) fluorescein isothiocyanate (FITC)-conjugated antibody (Alomone Labs, Jerusalem, Israel) for 1 h at room temperature in the dark at 200 rpm in concentrations according to the manufacturer’s instructions (10 mg antibody to 10 6 cells).

Techniques: Incubation, Lysis, Flow Cytometry

Journal: Critical Care

Article Title: Erythrocyte P2X 1 receptor expression is correlated with change in haematocrit in patients admitted to the ICU with blood pathogen-positive sepsis

doi: 10.1186/s13054-018-2100-3

Figure Lengend Snippet: Haematocrit and haemoglobin levels and erythrocyte P2X 1 receptor expression. a Change in haematocrit (hct) in blood pathogen-positive patients with sepsis, between Emergency Department (ER) and ICU admissions and P2X 1 receptor expression on erythrocytes. The right panel shows blood pathogen-positive patients grouped by high or low expression of P2X 1 (Δhct/hour, p = 0.011). b Change in haematocrit in blood pathogen-negative patients with sepsis between ER and ICU admission was not correlated with P2X 1 receptor expression on erythrocytes. The right panel shows erythrocyte turnover rate (Δhct/hour) in high P2X 1 -expressing or low P2X 1 -expressing patients. c Change in haemoglobin in blood pathogen-positive patients with sepsis between ER and ICU admissions and P2X 1 receptor expression on erythrocytes. The right panel shows the change in haemoglobin (ΔHgb/hour) in blood pathogen-positive patients grouped by high or low expression of P2X 1 ( p = 0.0375). d Change in haemoglobin from ER-admission to ICU in blood pathogen-negative patients with sepsis and P2X 1 receptor expression on erythrocytes. The right panel shows change in haemoglobin (ΔHgb/hour) between high P2X 1 -expressing and low P2X 1 -expressing patients. Data presented as mean ± SEM

Article Snippet: Cells in each sample were counted on a cell counter (Spectre, Merch Millipore, USA) prior to exposure to either P2X 1 (catalogue number APR-022-AG) or P2X 7 (catalogue number APR-008-F) fluorescein isothiocyanate (FITC)-conjugated antibody (Alomone Labs, Jerusalem, Israel) for 1 h at room temperature in the dark at 200 rpm in concentrations according to the manufacturer’s instructions (10 mg antibody to 10 6 cells).

Techniques: Expressing

Journal: Critical Care

Article Title: Erythrocyte P2X 1 receptor expression is correlated with change in haematocrit in patients admitted to the ICU with blood pathogen-positive sepsis

doi: 10.1186/s13054-018-2100-3

Figure Lengend Snippet: Correlation statistics

Article Snippet: Cells in each sample were counted on a cell counter (Spectre, Merch Millipore, USA) prior to exposure to either P2X 1 (catalogue number APR-022-AG) or P2X 7 (catalogue number APR-008-F) fluorescein isothiocyanate (FITC)-conjugated antibody (Alomone Labs, Jerusalem, Israel) for 1 h at room temperature in the dark at 200 rpm in concentrations according to the manufacturer’s instructions (10 mg antibody to 10 6 cells).

Techniques: Activity Assay, Expressing

Journal: Critical Care

Article Title: Erythrocyte P2X 1 receptor expression is correlated with change in haematocrit in patients admitted to the ICU with blood pathogen-positive sepsis

doi: 10.1186/s13054-018-2100-3

Figure Lengend Snippet: Haematocrit and haemoglobin levels and erythrocyte P2X receptor expression in patients infected with cytolysin-producing bacteria. a Change in haematocrit between Emergency Department (ER) and ICU admissions in patients with a positive blood culture result for cytolysin-producing bacteria and correlation with P2X 1 receptor expression on erythrocytes. b Change in haemoglobin between ER and ICU admissions in patients with a positive blood culture result for cytolysin-producing bacteria and correlation with P2X 1 receptor expression

Article Snippet: Cells in each sample were counted on a cell counter (Spectre, Merch Millipore, USA) prior to exposure to either P2X 1 (catalogue number APR-022-AG) or P2X 7 (catalogue number APR-008-F) fluorescein isothiocyanate (FITC)-conjugated antibody (Alomone Labs, Jerusalem, Israel) for 1 h at room temperature in the dark at 200 rpm in concentrations according to the manufacturer’s instructions (10 mg antibody to 10 6 cells).

Techniques: Expressing, Infection

Journal: Critical Care

Article Title: Erythrocyte P2X 1 receptor expression is correlated with change in haematocrit in patients admitted to the ICU with blood pathogen-positive sepsis

doi: 10.1186/s13054-018-2100-3

Figure Lengend Snippet: Haematocrit and haemoglobin levels and erythrocyte P2X 7 receptor expression. a Change in haematocrit in blood pathogen-positive patients with sepsis between Emergency Department (ER) and ICU admission and P2X 7 receptor expression on erythrocytes were not correlated. The right panel shows the change in haematocrit in blood pathogen-positive patients grouped by high or low expression of P2X 7 (Δhct/hour). b Change in haematocrit in blood pathogen-negative patients with sepsis between ER and ICU admission and P2X 7 receptor expression on erythrocytes. Right panel shows the change in haematocrit in blood pathogen-negative patients grouped by high or low expression of P2X 7 (Δhct/hour). c Change in haemoglobin in blood pathogen-positive patients with sepsis between ER and ICU admission and P2X 7 receptor expression on erythrocytes were not correlated. The right panel shows change in haemoglobin in blood pathogen-positive patients grouped by high or low expression of P2X 7 (ΔHgb/hour, not significant (ns)). d Change in haemoglobin in blood pathogen-negative patients with sepsis between ER and ICU admission and P2X 7 receptor expression on erythrocytes. The right panel shows change in haemoglobin in blood pathogen-negative patients grouped by high or low expression of P2X 7 (ΔHgb/hour)

Article Snippet: Cells in each sample were counted on a cell counter (Spectre, Merch Millipore, USA) prior to exposure to either P2X 1 (catalogue number APR-022-AG) or P2X 7 (catalogue number APR-008-F) fluorescein isothiocyanate (FITC)-conjugated antibody (Alomone Labs, Jerusalem, Israel) for 1 h at room temperature in the dark at 200 rpm in concentrations according to the manufacturer’s instructions (10 mg antibody to 10 6 cells).

Techniques: Expressing