phosphate buffered saline pbs  (Thermo Fisher)

Journal: Military Medical Research

Article Title: The cellular response capacity (CRC) as a novel immunomonitoring approach in sepsis

doi: 10.1016/j.mmr.2026.100010

Figure Lengend Snippet: Methodological workflow from sample preparation to measuring the cellular response capacity (CRC). Following blood collection, samples are prepared for flow cytometric analysis either unstimulated (PBS as buffer control) or stimulated with an inflammatory cocktail (cocktail of N-formylmethionyl-leucyl-phenylalanine, platelet-activating factor, and tumor necrosis factor). The CRC is calculated as the ratio of median fluorescence intensity (MFI) between stimulated and unstimulated neutrophils. Three approaches, classic, simple, and kinetic CRC, offer distinct advantages and limitations based on technical aspects such as manual processing steps (e.g., centrifugation) and incubation time. n =13–14. Statistical analysis was performed using the Kruskal-Wallis test with Dunn’s post-hoc test, comparing patients with sepsis at all time points (shown in the figure: 0 h, not shown in the figure: 24, 72, 120 h) with healthy volunteers (HV). P -values are indicated above the respective data points. Asterisks indicate significant differences between HV and patients at 0 h only. ⁎ P <0.05, ⁎⁎ P <0.01, ⁎⁎⁎ P <0.001. AUC. Area under the curve; AU. Arbitrary units; PBS. Phosphate-buffered saline; E. coli Escherichia coli .

Article Snippet: Subsequently, the monovettes were exposed to either phosphate-buffered saline (PBS) with calcium and magnesium (PBS +/+ , #14080055, Gibco, Thermo Fisher Scientific, Waltham, USA) and the bacterial culture media [buffer control of Escherichia coli ( E. coli ) suspension, hereafter referred to as BuC], viable E. coli bacteria (ATCC line 25922, DSMZ, Braunschweig, Germany), or LPS (100 ng/ml, from E. coli O55:B5, #L2637, Sigma Aldrich, Steinheim, Germany).

Techniques: Sample Prep, Control, Fluorescence, Centrifugation, Incubation, Saline

Journal: Military Medical Research

Article Title: The cellular response capacity (CRC) as a novel immunomonitoring approach in sepsis

doi: 10.1016/j.mmr.2026.100010

Figure Lengend Snippet: Comparison of different approaches to measure the cellular response capacity (CRC) for CD11b on neutrophil granulocytes. Blood from healthy volunteers was incubated with either PBS (Control) or 100 ng/ml LPS for 60 min in the ex vivo whole blood model. a Analysis of the median fluorescence intensity (MFI). b Evaluation of the CRC as determined by different approaches (classic, simple, and kinetic CRC). c, d Change in fluorescence intensity and change in CRC using the kinetic CRC approach, comparing blood with previous exposure to LPS or PBS (buffer control) from the ex vivo whole blood model. In both c and d , the single-cell values measured by flow cytometry were condensed using a moving median with a window of 9 cells. This moving median was then approximated with a 5 th -degree polynomial function. The plots display these polynomial functions together with the baseline (median before stimulation) and a connecting line from the baseline to the polynomial function for 30 s after stimulation. Values are shown as median and interquartile range. n= 10. Statistical analysis was performed using the Mann-Whitney U test. ⁎⁎⁎ P <0.001. AU. Arbitrary units; LPS. Lipopolysaccharide; PBS. Phosphate-buffered saline.

Article Snippet: Subsequently, the monovettes were exposed to either phosphate-buffered saline (PBS) with calcium and magnesium (PBS +/+ , #14080055, Gibco, Thermo Fisher Scientific, Waltham, USA) and the bacterial culture media [buffer control of Escherichia coli ( E. coli ) suspension, hereafter referred to as BuC], viable E. coli bacteria (ATCC line 25922, DSMZ, Braunschweig, Germany), or LPS (100 ng/ml, from E. coli O55:B5, #L2637, Sigma Aldrich, Steinheim, Germany).

Techniques: Comparison, Incubation, Control, Ex Vivo, Fluorescence, Single Cell, Flow Cytometry, MANN-WHITNEY, Saline

Journal: Journal of Sport and Health Science

Article Title: Weightlifting outperforms voluntary wheel running for improving adiposity and insulin sensitivity in obese mice

doi: 10.1016/j.jshs.2025.101100

Figure Lengend Snippet: Resistance exercise exceeds the benefits of endurance exercise in ameliorating metabolic dysfunction. Following 8 weeks of diet and exercise interventions, all mice were assessed for their metabolic function by GTT, ITT, and skeletal muscle response of Akt and AS160 phosphorylation to injection of insulin measured by Western blot. (A–C) HOMA-IR taken after an overnight fast for baseline glucose and insulin. (D and E) GTT from 0–120 min and calculated AUC; colored * indicates significant difference from NC-SED. (F and G) ITT from 0–60 min and calculated AUC; colored * indicates significant difference from NC-SED. (H–N) pAkt stimulation, AS160 S318, and AS160 T642 in hindlimb muscles before and after insulin injection and the pre–post ∆ in phosphorylation. (O and P) Total and phosphorylated 4E-BP1. (Q–T) Western results for Raptor, COX4, LC3 II/I, and ubiquitin staining. Representative western blot images inset right. Data presented as mean ± standard error of the mean. Statistical analysis performed by analysis of variance between groups: * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001. NC-SED n : 8–16 (white); HFD-SED n : 8–18 (red); HFD-R EX n : 8–16 (blue); HFD-E EX n : 8–15 (green). 4E-BP1 = Eukaryotic translation initiation factor 4E binding protein; Akt = protein kinase B; AS160 = Akt substrate 160; COX4 = cytochrome c oxidase 4; CS = citrate sythase; E EX = endurance exercise; GAPDH = glyceraldehyde 3 phosphate dehydrogenase; GTT = glucose tolerance test; HFD = high fat diet; HOMA-IR = homeostatic model assessment for insulin resistance; iAUC = integrated area under the curve; ITT = insulin tolerance test; LC3 II/I = microtubule-associated protein light chain 3; NC = normal chow; pAkt = phospho-Akt; R EX = resistance exercise; SED = sedentary; Ub = ubiquitin.

Article Snippet: Primary antibodies used for analysis were from Cell Signaling Technologies (Danvers, MA, USA) and diluted 1:1000 unless otherwise stated as follows: protein kinase B (Akt; 1:500; #4691; Cell Signaling Technologies), phospho-Akt (pAkt) S473 (1:500; #9271; Cell Signaling Technologies), Ubiquitin (#3933; Cell Signaling Technologies), microtubule-associated protein light chain 3 (LC3 II/I; #4018; Cell Signaling Technologies), cytochrome c oxidase subunit 4(COX4; #11967; Cell Signaling Technologies), Akt substrate 160 (AS160 S318; #8619; Cell Signaling Technologies), AS160 T642 (#8881; Cell Signaling Technologies), eukaryotic translation initiation factor 4E binding protein (4E-BP1; #9452; Cell Signaling Technologies), and glyceraldehyde 3 phosphate dehydrogenase (GAPDH; #2118; Cell Signaling Technologies).

Techniques: Phospho-proteomics, Injection, Western Blot, Muscles, Ubiquitin Proteomics, Staining, Binding Assay