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

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

Tenascin-C is a driver of inflammation in the DSS model of colitis

Inflammatory Bowel Disease (IBD) is a grouping of chronic inflammatory disorders of the gut. Tenascin-C is a pro-inflammatory, extracellular matrix protein found upregulated in IBD patients and whilst a pathological driver of chronic inflammation, its precise role in the etiology of IBD is unknown. To study tenascin-C’s role in colitis pathology we investigated its expression in a murine model of IBD. Wild-type (WT) or tenascin-C knockout (KO) male mice were left untreated or treated with dextran sodium sulphate (DSS) in their drinking water. Tenascin-C was upregulated at the mRNA level in the colitic distal colon of day eight DSS treated mice, coinciding with significant increases in gross and histological pathology. Immunohistochemistry localized this increase in tenascin-C to areas of inflammation and ulceration in the mucosa. Tenascin-C KO mice exhibited reduced gross pathology in comparison. These differences also extended to the histopathological level where reduced colonic inflammation and tissue damage were found in KO compared to WT mice. Furthermore, the severity of the distal colon lesions were less in the KO mice after 17 days of recovery from DSS treatment. This study demonstrates a role for tenascin-C as a driver of inflammatory pathology in a murine model of IBD and thus suggests neutralizing its pro-inflammatory activity could be explored as a therapeutic strategy for treating IBD

The extent of placental pathology is negatively correlated to birth weight in ewes infected with the wild-type strain of Chlamydia abortus

The placenta is the organ that allows the exchange of oxygen and nutrients between maternal and foetal blood, supplying the requirements of the growing foetus. Consequently, any factor that alters placental integrity may affect foetal nutrition, viability and lamb birth weight. Reproductive diseases, such as ovine enzootic abortion (OEA), impact on foetal viability due to the reduction in the functional area for maternofoetal exchange. This study aimed to consider the impact of pathological features of OEA placental lesions on lamb birth weight and indirectly on foetal viability. To investigate the relationship between birth weight and various OEA-related parameters, data from 562 lambs/foetuses from animals experimentally challenged with Chlamydia abortus strain S26/3 and from uninfected animals were analysed. The parameters investigated included the number of foetuses/lambs delivered (single/multiple), foetus/lamb sex, length of gestation, the proportion of placentas affected by lesions (percentage of gross placental pathology), foetal viability (live/aborted) and the number of C. abortus organisms shed in post-parturition vaginal excretions. The results suggest that the length of gestation and the proportion of placentas affected by lesions are the main contributors to birth weight variability, whereas the other factors, including foetal viability (live or aborted outcomes), were found to be less relevant co-variables. The study determined the strongest positive and negative correlations between birth weight were with the length of gestation and the extent of placental pathology, respectively. These results may indicate that economic losses associated with OEA infections result not only from aborted foetuses but also from the surviving lambs that are born weaker and consequently are more susceptible to diseases.EEA MercedesFil: Caspe, Sergio Gaston. Moredun Research Institute; Reino UnidoFil: Caspe, Sergio Gaston. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Mercedes; ArgentinaFil: Caspe, Sergio Gaston. University of Edinburgh. Royal (Dick) School of Veterinary Studies; Reino UnidoFil: Palarea-Albaladejo, Javier. Biomathematics & Statistics Scotland; Reino UnidoFil: Palarea-Albaladejo, Javier. University of Girona. Deparment of Computer Sciences, Applied Mathematics and Statistics; EspañaFil: Livingstone, Morag. Moredun Research Institute; Reino UnidoFil: Wattegedera, Sean Ranjan. Moredun Research Institute; Reino UnidoFil: Milne, Elspeth. University of Edinburgh. Royal (Dick) School of Veterinary Studies; Reino UnidoFil: Sargison, Neil Donald. University of Edinburgh. Royal (Dick) School of Veterinary Studies; Reino UnidoFil: Longbottom, David. Moredun Research Institute; Reino Unid