Association of adrenocorticotrophin and cortisol concentrations with peripheral blood leukocyte cytokine gene expression in septic and nonseptic neonatal foals

Background

The hypothalamic‐pituitary‐adrenal (HPA) is influenced by the proinflammatory cytokines IL‐6, IL‐1β, and TNF‐α in critically ill humans. Information about the association of cytokines with the HPA axis in neonatal foals is lacking.

Hypothesis/Objectives

The objectives were to describe for hospitalized septic and nonseptic foals (1) temporal changes in blood concentrations of ACTH, and cortisol, and leukocyte cytokine gene expression, and (2) coassociation of these HPA axis hormones with blood leukocyte cytokine gene expression.

Animals

Hospitalized septic foals (N = 15) and hospitalized nonseptic foals (N = 11).

Methods

Blood samples, obtained from study foals at admission (T = 0), and 24 (T = 1), 48 (T = 2), 72 (T = 3), and 96 (T = 4) hours after admission, were processed to isolate RNA from leukocytes and to harvest plasma and serum for hormone assays. Plasma ACTH and serum cortisol concentrations were determined by radioimmunoassay. Leukocyte mRNA expression of IL‐1β IL‐6, IL‐8, IL‐10, and TNF‐α was determined using RT‐PCR.

Results

Cortisol concentrations were greater (P < .05) in foals at admission than at other time points. The expressions of IL‐8 and IL‐10 mRNA were lower (P < .05) at each time point in septic than in nonseptic foals. Among septic foals, ACTH was positively associated (P = .0026) with IL‐6 mRNA expression.

Conclusions

Sepsis influences secretion of the HPA axis hormones and expression of cytokines in foals. A positive association with the HPA axis and IL‐6 expression was detected. The clinical importance of these findings requires additional study.     

Evaluation of disease activity markers in dogs with idiopathic inflammatory bowel disease

OBJECTIVES: To evaluate the clinical utility of serum tumour necrosis factor-alpha, C-reactive protein and microalbuminuria as disease activity markers in canine idiopathic inflammatory bowel disease. METHODS: Dogs with chronic gastrointestinal disease for which no underlying cause could be identified were considered to have idiopathic inflammatory bowel disease and were included in the study. Serum tumour necrosis factor-alpha was assessed using a canine-specific ELISA, C-reactive protein by immunoturbidometric assay and quantitative microalbuminuria was analysed using a monoclonal antibody directed against canine albumin. The canine inflammatory bowel disease activity index and histopathologic grade were used to assess disease severity; biologic markers were then compared with the canine inflammatory bowel disease activity index and histopathologic grade. RESULTS: Sixteen dogs were included in the study. C-reactive protein level was mildly elevated in 15 dogs. Microalbuminuria was elevated in two of 15 dogs, and tumour necrosis factor-alpha was not detected in any dog tested. No correlation was found between the canine inflammatory bowel disease activity index and C-reactive protein or microalbuminuria or between histopathologic grade and C-reactive protein or microalbuminuria. There was no correlation between histopathologic grade and the canine inflammatory bowel disease activity index. CLINICAL SIGNIFICANCE: Although only a small number of dogs were evaluated, this study does not support the use of serum tumour necrosis factor-alpha measured by canine-specific ELISA or microalbuminuria in the evaluation of disease activity in dogs with idiopathic inflammatory bowel disease. Although mildly elevated in most dogs, C-reactive protein did not reflect disease severity as assessed by the canine inflammatory bowel disease activity index or histopathologic grade.     

Mastocytemia in dogs with acute inflammatory diseases

Nineteen dogs were identified that had mastocytemia (mast cells in venous blood samples) not associated with mast cell neoplasia. The first 10 dogs were identified by examination of blood films from dogs with suspected parvovirus enteritis (8), fibrinous pericarditis and pleuritis secondary to thoracic lacerations (1), and renal insufficiency of unknown cause (1). Because of the apparent association with acute enteritis, blood films from 52 suspected canine parvovirus cases were examined retrospectively and 8 mastocytemic dogs were found. An additional 52 canine blood films were randomly selected from the same retrospective time period and 1 mastocytemic dog was found that had pneumothorax, pelvic fractures, and hemorrhagic septic abdominal effusion secondary to renal hemorrhage and traumatized intestines. All mastocytemic dogs had acute inflammatory leukograms the day that mast cells were first detected: neutropenia with toxic neutrophils (4), neutropenia with a left shift (8), total neutrophil count within reference interval but with a left shift (5), or neutrophilia with a left shift (2). All dogs except the renal insufficiency case had circulating toxic neutrophils. Five dogs were mastocytemic on more than 1 day. The pathogenesis of the mastocytemia associated with the acute inflammatory disease was not determined.     

Changes in leukocyte population after ozonated autohemoadministration in cows with inflammatory diseases

In this study, we investigated whether ozonated autohemoadministration (OAHA) influences leukocyte populations in cows with clinical inflammatory disease. Eleven cows with inflammatory disease (Inflammatory Group) and three healthy cows (Control Group) were used for this study. The CD4(+)/CD8(+) ratio in the Inflammatory Group increased significantly compared to that in the Control Group 3 to 4 days after OAHA treatment. In the Inflammatory Group, the number of CD14(+) cells decreased gradually after OAHA, but CD14(+) levels remained stable in the Control Group. The number of MHC class-II(+) cells decreased gradually in the Inflammatory Group, but increased gradually in the Control Group, and the difference between the groups was significant on day 14 after OAHA. These findings suggest a possible difference in the activation of immune response after OAHA in infected cows compared to healthy cows.     

Neutrophilic myeloperoxidase index and mean light absorbance in neonatal septic and nonseptic foals

Background: Two neutrophilic indices reported by the ADVIA 120 Hematology Analyzer, neutrophilic myeloperoxidase index (MPXI), and mean light absorbance (neutrophil X mean [NXM]) have been proposed as indicators of systemic inflammatory disease in horses and of neutrophil activation in coronary ischemic syndromes in people. Objective: The aim of this study was to evaluate NXM and MPXI in healthy, sick nonseptic, and sick septic foals to determine whether conditions likely associated with neutrophil activation result in decreases in these variables. Methods: In this retrospective study, CBC data from 61 neonatal foals presented to the Equine Teaching Hospital of Barcelona were evaluated for correlations between MPXI, NXM, percentage of large unstained cells, neutrophil count, and percentage of band neutrophils. Results obtained in septic (n=32), sick nonseptic (n=22), and healthy foals (n=7) were compared. In addition, results recorded in septic/neutropenic (n=12), septic/non‐neutropenic (n=20), nonseptic/neutropenic (n=8), nonseptic/non‐neutropenic (n=14), and healthy foals (n=7) were also compared. Results: A weak negative correlation was found between MPXI and neutrophil count and between NXM and percentage of band neutrophils. Septic/neutropenic foals had significantly higher MPXI values (median 17.9, minimum–maximum 4.7–42.5) than did septic/non‐neutropenic (1.5, ?24.4 to 22.3), nonseptic/neutropenic (6.6, 0.6–17.9), and nonseptic/non‐neutropenic foals (8.8, ?10.1 to 16.8) but did not differ significantly from controls (12.8, ?8.5 to 20.4). Conclusions: Significant differences in NXM or MPXI were not found when disease groups were compared with controls; however, septic/neutropenic foals had significantly higher median MPXI than other groups of sick foals. Further prospective studies are needed to clarify if this finding is related to decreased neutrophil function or activation in septic/neutropenic foals.     

Evaluation of lymphocyte apoptosis in dogs with inflammatory bowel disease

Objective-To determine whether lymphocyte apoptosis in intestinal mucosae is more common in healthy dogs than dogs with inflammatory bowel disease (IBD) and whether numbers of apoptotic cells increase after successful treatment of affected dogs. Animals-8 dogs with IBD (IBD dogs) and 8 healthy control dogs. Procedures-Biopsy specimens of the duodenum and colon were obtained via endoscopy from dogs with IBD before and after 10 weeks of standard treatment and compared with specimens obtained from control dogs. Expression of activated caspase 3 (Casp3), caspase-cleaved fragment p85 from poly-ADP-ribose polymerase (PARP), and B-cell leukemia/lymphoma 2 (Bcl-2) was measured in the duodenal (villous tip and base) and colonic mucosae. Results-Expression of Casp3 was greater in the duodenal villous tips of control dogs, compared with expression in similar tissues from dogs with IBD before or after treatment. Despite clinical improvement of dogs with IBD, expression of Casp3 did not increase after treatment. Expression of PARP did not differ between groups at any time point. Expression of Bcl-2 was greater at all 3 tissue sites in control dogs, compared with expression at the same sites in dogs with IBD. Furthermore, Bcl-2 expression in duodenal villous tips was higher in dogs with IBD after treatment but was not higher elsewhere. In control dogs, expression patterns for all 3 markers were similar between sites (villous tip > villous base > colon). Conclusions and Clinical Relevance-Expression of Casp3 in lymphocytes in duodenal villous tips was significantly reduced in dogs with IBD, compared with expression in healthy dogs, but no increase was detected following successful treatment of IBD. Increased expression of Bcl-2 may be a potential marker of the success of treatment.     

Changes in peripheral leukocyte subsets in dairy cows with inflammatory diseases after calving

To clarify the cellular immune system in dairy cows with inflammatory diseases after calving, the leukocyte subsets were examined in Holstein dairy cows. Twenty Holstein cows reared in one herd, were used in this study. Nine cows (Group 1) experienced onset of mastitis or puerperal fever within 2 weeks after calving, and the other eleven cows remained healthy (Group 2) after calving. The numbers of CD3(+), CD4(+) and CD8(+) cells tended to be lower in Group 1 than in Group 2 from the day of calving through week 1. These results suggested that the cows with inflammatory diseases might have experienced a decline in T cells by the day of calving, before the onset of disease.     

A comparison of five different bone resorption markers in osteosarcoma-bearing dogs, normal dogs, and dogs with orthopedic diseases

BACKGROUND: Various bone resorption markers in humans are useful for supporting the diagnosis of malignant skeletal pathology, with certain bone resorption markers appearing to be more discriminatory for detecting cancer-induced osteolysis than others. Canine osteosarcoma (OSA) is characterized by focal bone destruction, but a systematic investigation for determining which bone resorption marker best supports the diagnosis of OSA in dogs has not been reported. HYPOTHESIS: Dogs with OSA will have increased concentrations of bone resorption markers compared with healthy dogs and dogs with orthopedic disorders. Differences will exist among various bone resorption markers for their ability to support the diagnosis of malignant osteolysis in dogs with OSA. ANIMALS: Single time point, cross-sectional, cohort study including dogs with OSA (n = 20) or orthopedic disorders (n = 20) and healthy dogs (n = 22). METHODS: Basal concentrations of urine and serum N-telopeptide (NTx), urine and serum C-telopeptide (CTx), and urine deoxypyridinoline (DPD) were compared among all 3 groups. RESULTS: Compared with healthy dogs and dogs with orthopedic disorders, urine NTx, serum NTx, and serum CTx concentrations were significantly increased in dogs with OSA. For urine NTx and serum NTx, the calculated lower and upper 95% confidence limits in dogs with OSA did not overlap with dogs diagnosed with orthopedic disorders or healthy dogs. CONCLUSIONS AND CLINICAL IMPORTANCE: Of the markers evaluated in this study, urine NTx and serum NTx appear to be the most discriminatory resorption markers supporting the diagnosis of focal malignant osteolysis in dogs with OSA.     

Insulin-like growth factor I concentration in dogs with inflammatory and neoplastic liver diseases

Liver diseases are known to influence the serum concentration of insulin-like growth factor I (IGF-I) in humans, but such an effect has rarely been investigated in dogs. The aim of this study was to investigate serum IGF-I concentrations in dogs with primary liver diseases, in comparison with levels in healthy dogs and dogs with non-hepatic diseases. For this purpose, IGF-I serum concentrations were measured (DSL-5600 kit) in 36 dogs with various liver diseases and compared with 22 healthy controls and 20 dogs with non-hepatic diseases. The results showed that dogs with liver diseases had significantly lower IGF-I serum concentrations (P < 0.001) than clinical healthy dogs or dogs with non-hepatic diseases. But the results also indicate that the aetiology of liver disease has no influence on IGF-I serum concentration.     

Tenoscopy in 33 horses with septic and nonseptic digital tenosynovitis (1997–2001)

Tenoscopy is the use of an arthroscope to access tendon sheaths. This article reports clinical findings, treatments, and outcomes of 33 horses with either septic or nonseptic digital tenosynovitis submitted to 36 tenoscopies during 34 occasions at the Marion duPont Scott Equine Medical Center (1997–2001). Two of the 16 horses with septic tenosynovitis were euthanized at the hospital. From the 32 horses discharged from the hospital, outcome was obtained in 31 cases: 12 horses (6 septic) could work at the same level. No difference was detected between the outcome groups regarding clinical and laboratory variables. In nonseptic cases, lameness grade, circulating white blood cells, and plasma fibrinogen before surgery were lower, while the period between the onset of clinical signs and surgery was longer and duration of phenylbutazone treatment and hospitalization was shorter. No difference in the outcome was detected when septic and nonseptic cases were compared. Tenoscopy was useful for diagnosis and treatment of nonseptic and septic digital tenosynovitis. The small number of cases and the lack of accurate information due to the retrospective nature of the study may have prevented detection of a difference between the outcomes of horses with septic versus nonseptic tenosynovitis.     

Increased blood mRNA expression of inflammatory and anti-fibrotic markers in dogs with congestive heart failure

Inflammation and extracellular matrix (ECM) remodeling contribute to the development of congestive heart failure (CHF), but the pathogenesis is still incompletely understood. Therefore, whole blood samples from eight dogs without cardiac disease and eight dogs with CHF were investigated for mRNA expression of IL1β, IL2, IL4, IL6, IL8, IL10, TNFα, IFNγ, TGFβ1-3, MMP1, -2, -3, -9 and TIMP1-4 using quantitative PCR. Dogs with CHF had significantly higher IL1β (P=0.015), IL2 (P=0.043), MMP1 (P=0.031), TIMP3 (P=0.012) and lower TNFα (P<0.001), TGFβ3 (P=0.006), TIMP1 (P=0.015) and TIMP2 (P=0.011) mRNA levels. Increased pro-inflammatory IL1β and anti-fibrotic MMP1 and reduced pro-fibrotic TGFβ and TIMP1 and TIMP2 in dogs with CHF suggest progressive left ventricular remodeling. The reduction of TNFα and increase of immunomodulatory IL2 and TIMP3 might suggest control of the inflammatory response. A better understanding of inflammation and ECM remodeling in cardiac diseases may lead to novel treatment approaches.     

Pretreatment leukocyte ratios and concentrations as predictors of outcome in dogs with cutaneous mast cell tumours

Leukocyte ratios correlate with outcome in several human cancers. Little is known about their prognostic significance in mast cell tumour (MCT). The aim of the study was to evaluate the prognostic significance of pretreatment leukocyte concentrations and their ratios in dogs with MCT for survival. Medical records of 92 dogs with MCT were retrospectively reviewed. Tumour diagnosis was made by tumour biopsy or fine‐needle aspirate. Only dogs without prior treatment were included. Eosinophil, lymphocyte, monocyte and neutrophil concentration were obtained by ADVIA 2120? (Siemens Healthcare, Vienna, Austria). Neutrophil‐to‐eosinophil ratio (NER), lymphocyte‐to‐monocyte ratio (LMR) and neutrophil‐to‐lymphocyte ratio (NLR) were calculated from collected leukocyte concentrations. Relative eosinophil concentration (REC), NER (P < 0.001), NLR (P = 0.001) and LMR (P < 0.001) were significant prognostic factors for outcome in univariate analysis. REC (P = 0.008) and NER (P = 0.001) remained independent predictors of survival in multiple analyses. Leukocyte concentrations and ratios, especially REC and NER may serve as prognostic indicators in MCT.     

Evaluation of plasma inflammatory cytokine concentrations in racing sled dogs

In human athletes significant changes in cytokine concentrations secondary to exercise have been observed. This prospective study evaluated the effect of a multi-day stage sled dog race on plasma concentrations of monocyte chemoattractant protein-1 (MCP-1), tumor necrosis factor-alpha (TNF-α), interleukin-2 (IL-2), interleukin-6 (IL-6), interleukin-8 (IL-8), and interleukin-10 (IL-10). Samples from 20 dogs were harvested prior to and on days 2 and 8 of an 8-day race. Exercise resulted in significantly decreased TNF-α and IL-8 as well as increases of MCP-1, IL-6, and IL-10 concentrations (P-value between 0.01 and < 0.0001 for all parameters). The proportion of values for IL-2 that were below the detection limit increased from 40% on day 0 to 75% on day 2 and decreased on day 8 to 40% (P = 0.04). Racing sled dogs show cytokine-concentration changes that are different from those in humans.