A New Cecal Slurry Preparation Protocol with Improved Long-Term Reproducibility for Animal Models of Sepsis

Starr, Marlene E.; Steele, Allison M.; Saito, Mizuki; Hacker, Bill J.; Evers, B. Mark; Saitō, Hiroshi

doi:10.1371/journal.pone.0115705

Cited by 155 publications

(159 citation statements)

References 31 publications

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“…A small ventral incision on the neck was made, and the left common carotid artery was isolated prior to cannulation. The catheter was advanced until placed in the aortic arch and secured in place with sutures [26]. The transmitter body was then advanced subcutaneously to lie over the right flank before the incision was closed.…”

Section: Mouse Blood Pressure Telemetrymentioning

confidence: 99%

The response of the innate immune and cardiovascular systems to LPS in pregnant and nonpregnant mice†

Zöllner

Howe

Edey

et al. 2017

Biology of Reproduction

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Sepsis is the leading cause of direct maternal mortality, but there are no data directly comparing the response to sepsis in pregnant and nonpregnant (NP) individuals. This study uses a mouse model of sepsis to test the hypothesis that the cardiovascular response to sepsis is more marked during pregnancy. Female CD1 mice had radiotelemetry probes implanted and were time mated. NP and day 16 pregnant CD-1 mice received intraperitoneal lipopolysaccharide (LPS; 10 μg, serotype 0111: B4). In a separate study, tissue and serum (for RNA, protein and flow cytometry studies), aorta and uterine vessels (for wire myography) were collected after LPS or vehicle control administration. Administration of LPS resulted in a greater fall in blood pressure in pregnant mice compared to NP mice. This occurred with similar changes in the circulating levels of cytokines, vasoactive factors, and circulating leukocytes, but with a greater monocyte and lesser neutrophil margination in the lungs of pregnant mice. Baseline markers of cardiac dysfunction and apoptosis as well as cytokine expression were higher in pregnant mice, but the response to LPS was similar in both groups as was the ex vivo assessment of vascular function. In pregnant mice, nonfatal sepsis is associated with a more marked hypotensive response but not a greater immune response. We conclude that endotoxemia induces a more marked hypotensive response in pregnant compared to NP mice. These changes were not associated with a more marked systemic inflammatory response in pregnant mice, although monocyte lung margination was greater. The more marked hypotensive response to LPS may explain the greater vulnerability to some infections exhibited by pregnant women. Pregnancy enhances LPS-induced hypotension, 2017, Vol. 97, No. 2 Summary SentenceIn pregnant mice, nonfatal sepsis is associated with a more marked hypotensive response but not a greater immune response.

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Section: Mouse Blood Pressure Telemetrymentioning

confidence: 99%

The response of the innate immune and cardiovascular systems to LPS in pregnant and nonpregnant mice†

Zöllner

Howe

Edey

et al. 2017

Biology of Reproduction

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“…A cecal slurry was created by harvesting the cecal contents of a sacrificed adult mouse and preparing an injectable solution according to a previously validated protocol. [6] This slurry was intraperitoneally injected into breastfed age-matched mice of the same strain, and BOS measurements were subsequently obtained.…”

Section: Methodsmentioning

confidence: 99%

Early detection of necrotizing enterocolitis using broadband optical spectroscopy

Goldstein¹,

Beaulieu²,

Niño³

et al. 2018

Journal of Pediatric Surgery

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Purpose: The definitive diagnosis of necrotizing enterocolitis (NEC) is typically at an advanced stage, indicating the need for sensitive and noninvasive diagnostic modalities. Near infrared spectroscopy (NIRS) has been utilized to noninvasively measure intraabdominal tissue oxygenation and to diagnose NEC, but specificity is lacking, in part because sensors are limited to a narrow band of the electromagnetic spectrum. Here, we introduce the concept of broadband optical spectroscopy (BOS) as a noninvasive method to characterize NEC. Methods: NEC was induced in 7-day old mice by gavage feeding with formula supplemented with enteric bacteria plus hypoxia. Transabdominal spectroscopy was performed daily using a broad-spectrum halogen light source coupled with a spectroradiometer capable of detection from 400 to 1800 nm. Results: A feature extraction algorithm was developed based on the spectral waveforms from mice with NEC. When subsequently tested on cohorts of diseased and control mice by a blinded examiner, noninvasive BOS was able to detect disease with 100% specificity and sensitivity. Conclusions: We reveal that the use of BOS is able to accurately and noninvasively discriminate the presence of NEC in a mouse model, thus introducing a noninvasive early diagnostic modality for this devastating disease.

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“…injection with the minimum lethal dose (LD 100 ) of cecal slurry (CS), prepared as we recently described in detail (9). Acute systemic inflammation was induced by injection with lipopolysaccharide (LPS) derived from Pseudomonas aeruginosa (Sigma-Aldrich, St. Louis, MO).…”

Section: Methodsmentioning

confidence: 99%

Short-Term Dietary Restriction Rescues Mice From Lethal Abdominal Sepsis and Endotoxemia and Reduces the Inflammatory/Coagulant Potential of Adipose Tissue

Starr

Steele

Cohen

et al. 2016

Critical Care Medicine

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Objective Visceral adipose tissue is a major site for expression of pro-inflammatory and pro-coagulant genes during acute systemic inflammation. In this study, we tested whether loss of fat mass by dietary restriction (DR) would remove the major source of these factors resulting in improved tolerance to sepsis and endotoxemia. Design Prospective, laboratory controlled experiments. Setting Aging and Critical Care Research Laboratory in a university hospital. Subjects Middle-aged (12 month-old) male C57BL/6 mice. Interventions Mice were subjected to 40% DR for three weeks followed by induction of abdominal sepsis or endotoxemia by intraperitoneal injection with cecal slurry or lipopolysaccharide (LPS), respectively. Measurements and main results Compared to freely-fed mice, DR mice exhibited dramatically improved survival (80% vs 0% after sepsis, p<0.001;86% vs 12%after endotoxemia, p=0.013) and significantly reduced visceral fat-derived mRNA expression of interleukin-6 (IL-6), thrombospondin-1, plasminogen activator inhibitor-1, and tissue factor which positively correlated with fat mass. Plasma levels of IL-6 were significantly reduced by DR and correlated with adipose IL-6 mRNAlevels and fat mass (p<0.001, R2=0.64 and 0.89). In vitro culture of visceral fat explants from naïve DR mice showed significantly reduced IL-6 secretion compared to that from freely-fed mice in response to LPS. Analysis of major adipose immune cell populations by flow cytometry demonstrated that macrophages were the only cell population reduced by DR and that CD11c+/CD206+ (M2-type) and CD11c-/CD206- (double negative) macrophages, in addition to T cells, are the major immune cell populations that produce IL-6 in middle-age during systemic inflammation. Conclusions Short-term DR drastically improved the survival outcome of middle-aged mice during both polymicrobial sepsis and sterile endotoxemia. Improved survival was accompanied by a significantly attenuated inflammatory response in adipose tissue, which is likely due to alterations of both fat mass quantity and qualitative changes including a reduction in macrophage populations.

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A New Cecal Slurry Preparation Protocol with Improved Long-Term Reproducibility for Animal Models of Sepsis

Cited by 155 publications

References 31 publications

The response of the innate immune and cardiovascular systems to LPS in pregnant and nonpregnant mice†

The response of the innate immune and cardiovascular systems to LPS in pregnant and nonpregnant mice†

Early detection of necrotizing enterocolitis using broadband optical spectroscopy

Short-Term Dietary Restriction Rescues Mice From Lethal Abdominal Sepsis and Endotoxemia and Reduces the Inflammatory/Coagulant Potential of Adipose Tissue

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