Location, Location, Location

Abstract

Objective Considerable breakthroughs in the field of sepsis have been made using animal models. Sepsis exhibits a wide array of derangements that may be evaluated in the blood, including the release of pro- and anti-inflammatory cytokines. The Shock journal adheres to the ARRIVE guidelines regarding reporting in vivo results to allow reproducibility of data findings. It is generally assumed that blood cytokine concentrations collected from typical sampling sites will be similar, but there are no data validating that this is true. The main purpose of the present study was to determine if the location of blood sampling results in cytokine concentration differences following inflammatory insults. Methods Two different models of acute inflammation were studied. Adult, female ICR mice were injected with E. coli lipopolysaccharide (LPS, n = 28) or subjected to cecal ligation and puncture (CLP) (n = 16). They were sacrificed at early time points following these inflammatory challenges for the collection of blood from the facial vein, retro-orbital sinus, and heart. Additonal samples were collected in EDTA and heparin. Plasma cytokines from the same mouse were collected from each sampling site and evaluated by ELISA. Clinical chemical parameters including plasma BUN and total protein were also analyzed. Results Regardless of model, time of collection, or cytokine measured, cytokine values from heart blood were higher than facial vein values from the same mouse. IL-6 collected from the heart relative to the facial vein demonstrated elevated concentrations following injection of LPS. In a similar manner, higher concentrations of IL-6, MIP-2, IL-10 and IL-1RA were found in cardiac puncture samples compared to other sampling sites 24 hours following sepsis induced by cecal ligation and puncture. Similar differences were not seen when comparing BUN and total protein values from the two different sites. Using plasma IL-6 collected from the heart would incorrectly stratify predicted to live (Live-P) mice into the predicted to die (Die-P) category. Therefore, a simple linear regression model was developed to correctly re-stratify mice to their predicted fate. These data demonstrate that pro- and anti-inflammatory cytokine concentrations are dramatically elevated when drawn centrally from the heart compared to collection from peripheral locations such as the facial vein. It is critical for publications to document the sampling location when evaluating plasma cytokines and attempting to compare studies.

Assessment of the adjuvant activity of mesoporous silica nanoparticles in recombinant Mycoplasma hyopneumoniae antigen vaccines

“…Mice immunized only with each of the MSNs or with Alhydrogel 2% VacciGrade in the same conditions described above were used as immunization negative control groups. Blood samples were collected from the facial vein [24] at 0 day (preimmune negative control) and 15, 30 and 45 days after the first immunization (DAI). Euthanasia was carried out 60 DAI, prior to spleen removal.…”

Viral Nucleic Acids in the Serum Are Dependent on Blood Sampling Site in Patients with Clinical Suspicion of Myocarditis

“…The detection of viral nucleic acids in serum collected from different sites was not taken into the consideration in patients with suspected MCI or iDCM; however, it was demonstrated that the sampling place is a critical factor for determining a difference in inflammatory cytokine values [18]. In an experimental study, Mella et al [18] showed that the cytokines were dramatically elevated in ventricle blood compared with peripheral blood in infected mice.…”

“…In an experimental study, Mella et al [18] showed that the cytokines were dramatically elevated in ventricle blood compared with peripheral blood in infected mice. We suspected that this phenomenon can be possible in patients with MCI or/and iDCM not only regarding cytokines, but also viral nucleic acids.…”

Linoleic acid improves Piezo 2 dysfunction in a mouse model of Angelman syndrome

“…Mouse blood was obtained by cardiac puncture under general anesthesia, and immediately mixed with a 10% vol. of heparin (1000 units/ Article https://doi.org/10.1038/s41467-023-36818-0 ml; Sagent Pharmaceuticals), and spun down at 2,000 g for 15 min 106 . Then, the plasma was separated and used for the cytokine profile assay.…”