Differential white blood cell counts in rabbits: a comparison of the Advia 2120 and a manual method

We evaluated the performance of the Advia 2120 (Siemens) differential leukocyte count (A-Diff) compared to the manual method (M-Diff) in rabbits. EDTA-anticoagulated blood samples collected for diagnostic purposes were analyzed within 6 h of collection. The M-Diff was performed blindly by 2 observers on blood smears by counting 200 cells. We initially included 117 samples; 25 samples were excluded because of suboptimal gating of leukocytes in the Advia peroxidase cytogram or poor blood smear quality. The correlation between the A-Diff and M-Diff was very high for heterophils (r = 0.924, p < 0.001) and lymphocytes (r = 0.903, p < 0.001), high for basophils (r = 0.823, p < 0.001), moderate for monocytes (r = 0.645, p < 0.001), and low for eosinophils (r = 0.336, p = 0.001). The Passing–Bablok regression analyses revealed a small-to-moderate constant error for lymphocytes and a slight constant error for basophils. Small proportional errors were detected for heterophils, lymphocytes, and eosinophils. The Bland–Altman analyses revealed that the Advia significantly underestimates heterophils and overestimates lymphocytes compared to M-Diff. The biases for the other leukocytes were minimal and likely clinical insignificant; however, our results, particularly for eosinophils, should be interpreted cautiously given the observed low percentages in our samples. Given the observed biases in heterophil and lymphocyte percentages in the Advia 2120 CBC results in rabbits, method-specific reference intervals should be used. The Advia can recognize leporine basophils. Evaluation of blood smears is still recommended to investigate abnormal results and erroneous cytograms reported by the Advia

Local lung responses following endobronchial elastase and lipopolysaccharide instillation in sheep

Chronic lipopolysaccharide (LPS) exposure may contribute to the pathogenesis of a number of lung diseases including COPD and emphysema. We sought to develop a large- animal model of emphysema using repeated LPS administration into sheep lung segments. An experimental protocol was designed to facilitate comparisons with elastase-treated and control segments within the same lung of individual sheep. Histopathologic evaluation of segments treated with LPS demonstrated low-grade inflammation characterized by an increase in the number of intra-alveolar macrophages and lymphocytes. Treated segments demonstrated a significant reduction in airspace surface area (ASA), an increase in percent disrupted alveolar attachments and the distance between normal alveolar attachments, and a reduction in the number of normal alveolar attachments surrounding nonrespiratory bronchioles. Coefficient of variation of individual ASA measurements in elastase-treated segments was indicative of a heterogeneous parenchymal response, in contrast to that associated with chronic LPS treatment. Our results demonstrate that chronic LPS treatment of individual lung segments in sheep induces microscopic emphysema qualitatively and quantitatively consistent with both accepted pathologic definitions of this condition and with that produced by airway instillation of elastolytic enzymes. Development of this phenotype is associated with evidence of downregulated activation of transforming growth factor beta

Suppressor of cytokine signaling 2 (SOCS2) deletion protects bone health of mice with DSS induced inflammatory bowel disease.

Individuals with inflammatory bowel disease (IBD) often present with poor bone health. The development of targeted therapies for this bone loss requires a fuller understanding of the underlying cellular mechanisms. Although bone loss in IBD is multifactorial the altered sensitivity and secretion of growth hormone (GH) and insulin-like growth factor-1 (IGF-1) in IBD is understood to be a critical contributing mechanism. The expression of suppressor of cytokine signaling 2 (SOCS2), a well-established negative regulator of GH signaling, is stimulated by pro-inflammatory cytokines. Therefore, it is likely that SOCS2 expression represents a critical mediator through which pro-inflammatory cytokines inhibit GH/IGF-1 signaling and decrease bone quality in IBD. Utilising the DSS model of colitis we have revealed that endogenously elevated GH function in the Socs2−/− mouse protects the skeleton from osteopenia. Micro-computed tomography assessment of DSS treated wild-type mice revealed a worsened trabecular architecture compared to control mice. Specifically, DSS treated WT mice had significantly decreased bone volume (BV/TV) (41%; p&#60;0.05), trabecular thickness (16%; p&#60;0.05), trabecular number (30%; p&#60;0.05), and a resulting increase in trabecular separation (19%; &#60;0.05). In comparison, the trabecular bone of Socs2 deficient mice was partially protected from the adverse effects of DSS. The reduction in a number of parameters including BV/TV (21%; p&#60;0.05) was less, and no changes were observed in trabecular thickness or separation. This protected phenotype was unlikely to be a consequence of improved mucosal health in the DSS treated Socs2−/− mice but rather a result of unregulated GH signaling directly on bone. These studies indicate that the absence of SOCS2 is protective against bone loss typical of IBD. This study also provides an improved understanding of the relative effects of GH/IGF-1 on bone health in experimental colitis, information that is essential before these drugs are explored as bone protective agents in children and adults with IBD

Deficiency of the bone mineralization inhibitor NPP1 protects against obesity and diabetes

The emergence of bone as an endocrine regulator has prompted a re-evaluation of the role of bone mineralization factors in the development of metabolic disease. Ectonucleotide pyrophosphatase/phosphodiesterase-1 (NPP1) controls bone mineralization through the generation of pyrophosphate, and levels of NPP1 are elevated both in dermal fibroblast cultures and muscle of individuals with insulin resistance. We investigated the metabolic phenotype associated with impaired bone metabolism in mice lacking the gene that encodes NPP1 (Enpp1−/− mice). Enpp1−/− mice exhibited mildly improved glucose homeostasis on a normal diet but showed a pronounced resistance to obesity and insulin resistance in response to chronic high-fat feeding. Enpp1−/− mice had increased levels of the insulin-sensitizing bone-derived hormone osteocalcin but unchanged insulin signalling within osteoblasts. A fuller understanding of the pathways of NPP1 could inform the development of novel therapeutic strategies for treating insulin resistance