A Retrospective Comparative Evaluation of Selected Blood Cell Ratios, Acute Phase Proteins, and Leukocyte Changes Suggestive of Inflammation in Cats.

Neutrophil-to-lymphocyte (NLR), monocyte-to-lymphocyte (MLR), and platelet-to-lymphocyte (PLR) ratios have been proposed as diagnostic and prognostic markers for neoplastic and inflammatory diseases in dogs and cats. The aim of this retrospective preliminary study was to evaluate the relationship between these ratios and markers of inflammation routinely measured in cats. A total of 275 cats were enrolled. Complete blood count, serum amyloid A (SAA), albumin, globulin, and albumin-to-globulin ratio (AGR) data were analyzed, as well as the presence of leukocyte alterations considered suggestive of inflammation (LAI: neutrophils left shift, toxic neutrophils, and reactive lymphocytes) evaluated in blood smears. The NLR and MLR correlated positively with SAA and globulins and negatively with albumin and AGR. Higher NLR and MLR were found in cats with increased SAA and globulins and decreased albumin and AGR. The PLR correlated negatively with albumin and AGR. A higher PLR was found in cats with hypoalbuminemia. Cats with LAI had higher NLR, MLR, and PLR. In cats with no changes in parameters indicative of inflammation, 11.25, 0.42, and 528.3 were identified as upper limits for NLR, MLR, and PLR, respectively. In conclusion, the NLR, MLR, and PLR act as good inflammatory markers easily evaluated by routine hematology.

Leptospira spp. Prevalence in Cats from Southern Italy with Evaluation of Risk Factors for Exposure and Clinical Findings in Infected Cats.

Leptospirosis is a worldwide zoonotic disease, but feline leptospirosis is rarely reported. This study aimed at investigating Leptospira spp. prevalence in cats from southern Italy, evaluating risk factors, clinical findings and laboratory data associated with infection. The serum of 112 cats was investigated by microscopic agglutination test (MAT), detecting anti-Leptospira antibodies against 14 pathogenic serovars. Blood and urine samples were tested by a real-time polymerase chain reaction targeting the lipL32 gene of pathogenic Leptospira. Antibodies against serovars Poi, Bratislava, Arborea, Ballum, Pomona and Lora were detected in 15.3% (17/111) of cats (titers range: 20-320). Leptospira spp. DNA was found in 3% (4/109) of blood and 9% (10/111) of urine samples. The spring season was the only risk factor for urinary Leptospira DNA shedding. Laboratory abnormalities significantly associated and/or correlated with Leptospira spp. positivity were anemia, monocytosis, neutrophilia, eosinopenia, increased alanine aminotransferase activity, hypoalbuminemia and hyperglobulinemia. In the investigated areas, cats are frequently infected by Leptospira spp. and can represent an additional reservoir or sentinel for a risk of infection. Moreover, some laboratory changes could be compatible with a pathogenic effect of Leptospira spp. in the feline host.

Retrospective evaluation of hyperproteinorrachia without pleocytosis (albuminocytologic dissociation) and survival in dogs.

BACKGROUND: Hyperproteinorrachia (raised cerebrospinal fluid total protein [CSF-TP]) without pleocytosis (HP) (also known as albuminocytologic dissociation) is identified in dogs with different neurologic diseases. However, the association between survival and increased CSF-TP is unknown. OBJECTIVES: (a) Identify conditions commonly associated with HP in dogs and (b) investigate whether higher CSF-TP concentrations or other relevant factors are associated with 1-year survival. METHODS: This is a retrospective study that identified dogs with HP (Cisternal CSF-TP >0.30 g/L, Lumbar CSF-TP >0.45 g/L with total nucleated cell concentrations [TNCCs] and RBC counts within RIs) from 2008 to 2019: recording signalment, weight, vital parameters, inflammation, neuroanatomic localization, CSF-TP, sampling site, final diagnosis, etiologic classification, and 1-year survival. Corrected CSF-TP was calculated as CSF-TP minus 0.3 (cisternal) or 0.45 (lumbar or unknown). Descriptive statistics were produced, CSF-TP differences between groups (eg, neuroanatomic localizations) were evaluated using the Mann-Whitney U test or Kruskal-Wallis test (post-hoc testing). The Cox proportional hazards model was used for survival data. Statistical significance was set at a P < 0.05. RESULTS: In all, 39 dogs had HP, associated with 17 conditions, including neoplasia (n = 6), meningoencephalitis of unknown origin (n = 4) (MUO), and intervertebral disc disease (n = 4) (IVDD) as the most common conditions. There was no significant difference between the CSF-TP/corrected CSF-TP between 1-year survivors and non-survivors, nor was there a difference between different neuroanatomic localizations or etiologic classifications (P > 0.05). Neoplasia, after adjustment for age, was the only variable associated with a worse survival (P = 0.01 HR: 2.08 (95% CI: 1.65-39.2). CSF-TP was not associated with age (P > 0.05). CONCLUSIONS: HP in dogs is associated with a wide range of conditions; the most common conditions are neoplasia, MUO, and IVDD. Higher CSF-TP levels do not correlate with a worse 1-year survival; however, they do correlate with neoplastic lesions.

Serum amyloid A levels and alpha 2 and gamma globulins on serum protein electrophoresis in cats exposed to and infected with Leishmania infantum.

BACKGROUND: Dogs are the main reservoir hosts of Leishmania infantum; nevertheless, recent investigations indicate a likely role for cats in the epidemiology of Leishmania infection. Feline leishmaniosis (FeL) remains poorly characterised, partly due to the lack of suitable diagnostic tools. This study aimed to compare serum amyloid A (SAA) levels and serum protein electrophoresis (SPE) profiles (specifically, alpha 2 and gamma globulins) in cats naturally exposed to or infected by L. infantum from southern Italy versus those of healthy controls and versus cats with neoplastic or inflammatory conditions from non-endemic areas. METHODS: Serum or plasma samples from four cohorts of cats were analysed for SAA levels and by SPE: (i) G1: healthy controls from Leishmania-non-endemic regions of Switzerland; (ii) G2: cats pre-diagnosed with neoplastic or inflammatory conditions available from the University of Cambridge sample archive; (iii) G3: L. infantum-seropositive, quantitative (q)PCR-negative cats from southern Italy; (iv) G4: L. infantum-seropositive and qPCR-positive cats from southern Italy. SAA data were assessed for normality and homoscedasticity using the Shapiro-Wilk and Levene's tests, respectively; the Kruskall-Wallis test, followed by Dunn's test with Bonferroni correction were subsequently used to compare SAA serum levels between groups. A weighted generalised linear model with a binomial distribution was used to assess statistically significant differences in the numbers of animals displaying elevated gamma globulins and increased alpha 2 globulins between groups. RESULTS: Overall, 68 samples were analysed (G1: n = 16, G2: n = 20, G3: n = 20, G4: n = 12). Cats suffering from neoplastic and inflammatory conditions (G2 ) showed significantly higher SAA levels than healthy controls (G1) (median values [interquartile range]: G1: 0.00 [0.00-0.00] mg/l versus G2: 0.85 [0.00-49.55] mg/l). G2, G3 and G4 cats showed higher percentages of individuals with increased alpha 2 globulins (percentages ± standard error: G1 = 20.0% ± 10.3, G2 = 80.0% ± 8.9, G3 = 70.0% ± 10.2, G4 = 75.0% ± 12.5) and gamma globulins (G1 = 0.0% ± 0, G2 = 65.0% ± 10.7, G3 = 50.0% ± 11.2, G4 = 58.3% ± 14.2) than healthy control cats (G1). For all three markers, no significant difference between cats within G2, G3 and G4 was recorded. CONCLUSIONS: This study indicates that the proportions of animals with elevated levels of alpha 2 and gamma globulins are significantly higher in cats exposed to and infected with L. infantum. Levels of SAA and alpha 2 and gamma globulins may not be used to differentiate between L. infantum infection or exposure, and neoplastic and/or inflammatory conditions.

Publisher Correction: In Utero Gene Therapy (IUGT) Using GLOBE Lentiviral Vector Phenotypically Corrects the Heterozygous Humanised Mouse Model and Its Progress Can Be Monitored Using MRI Techniques.

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

In Utero Gene Therapy (IUGT) Using GLOBE Lentiviral Vector Phenotypically Corrects the Heterozygous Humanised Mouse Model and Its Progress Can Be Monitored Using MRI Techniques.

In utero gene therapy (IUGT) to the fetal hematopoietic compartment could be used to treat congenital blood disorders such as ß-thalassemia. A humanised mouse model of ß-thalassemia was used, in which heterozygous animals are anaemic with splenomegaly and extramedullary hematopoiesis. Intrahepatic in utero injections of a ß globin-expressing lentiviral vector (GLOBE), were performed in fetuses at E13.5 of gestation. We analysed animals at 12 and 32 weeks of age, for vector copy number in bone marrow, peripheral blood liver and spleen and we performed integration site analysis. Compared to noninjected heterozygous animals IUGT normalised blood haemoglobin levels and spleen weight. Integration site analysis showed polyclonality. The left ventricular ejection fraction measured using magnetic resonance imaging (MRI) in treated heterozygous animals was similar to that of normal non-ß-thalassemic mice but significantly higher than untreated heterozygous thalassemia mice suggesting that IUGT ameliorated poor cardiac function. GLOBE LV-mediated IUGT normalised the haematological and anatomical phenotype in a heterozygous humanised model of ß-thalassemia.

Myoclonus and hypercalcemia in a dog with poorly differentiated lymphoproliferative neoplasia.

A 1-year, 8-month-old Rhodesian Ridgeback was presented with obtundation, ambulatory tetraparesis, and myoclonus. Initial clinical findings included ionized hypercalcemia with an apparent marked increase in parathyroid hormone, thrombocytopenia, and nonregenerative anemia. Low numbers of circulating atypical cells were noted on blood film evaluation. Brain magnetic resonance imaging identified an extra-axial contrast enhancing subtentorial lesion, and cerebrospinal fluid (CSF) analysis documented a marked atypical lymphocytic pleocytosis. Flow cytometry performed on the CSF demonstrated expression of only CD45, CD90, and MHC class II, with Pax5 positivity on subsequent immunohistochemistry. The final diagnosis was of B-cell lymphoblastic lymphoma or acute leukemia, given the distribution of disease and the presence of significant bone marrow infiltration alongside an aggressive clinical course. The unusual immunophenotype of the neoplastic cells and hypercalcemia presented antemortem diagnostic challenges, highlighting the need for a multidisciplinary approach and caution in the interpretation of clinical abnormalities in cases with multiple comorbidities.

Validation of a commercial 1,2-o-dilauryl-rac-glycero glutaric acid-(6'-methylresorufin) ester lipase assay for diagnosis of canine pancreatitis.

The objectives of this study were fourfold: technical validation of a commercial canine 1,2-o-dilauryl-rac-glycero glutaric acid-(6'-methylresorufin) ester (DGGR) lipase assay, to calculate a reference interval for DGGR lipase by the indirect a posteriori method, to establish biological validity of the assay, and to assess agreement between DGGR lipase and specific canine pancreatic lipase (Spec cPL) assays. Dogs with histologically confirmed acute pancreatitis (n=3), chronic pancreatitis (n=8) and normal pancreatic tissue (n=7) with stored (-80°C) serum samples were identified. Relevant controls were selected. Precision, reproducibility and linearity of DGGR lipase, and the effect of sample haemolysis and freezing, were assessed. Sensitivity and specificity of DGGR lipase and Spec cPL were determined. Agreement between these two parameters was calculated using Cohen's kappa coefficient (κ). The DGGR lipase assay demonstrated excellent precision, reproducibility and linearity. Sample haemolysis and storage at -80°C for 12 months did not influence the assay. DGGR lipase (>245IU/l) and Spec cPL (>400µg/l) both showed poor sensitivity but excellent specificity for acute pancreatitis, and poor to moderate sensitivity but excellent specificity for chronic pancreatitis. Substantial agreement (κ=0.679) was found between DGGR lipase and Spec cPL. The validated DGGR lipase assay had similar sensitivity and specificity for the diagnosis of acute and chronic pancreatitis to Spec cPL. DGGR lipase is a reliable alternative to Spec cPL for the diagnosis of pancreatitis.

Comprehensive annotation and evolutionary insights into the canine (Canis lupus familiaris) antigen receptor loci.

Dogs are an excellent model for human disease. For example, the treatment of canine lymphoma has been predictive of the human response to that treatment. However, an incomplete picture of canine (Canis lupus familiaris) immunoglobulin (IG) and T cell receptor (TR)-or antigen receptor (AR)-gene loci has restricted their utility. This work advances the annotation of the canine AR loci and looks into breed-specific features of the loci. Bioinformatic analysis of unbiased RNA sequence data was used to complete the annotation of the canine AR genes. This annotation was used to query 107 whole genome sequences from 19 breeds and identified over 5500 alleles across the 550 genes of the seven AR loci: the IG heavy, kappa, and lambda loci; and the TR alpha, beta, gamma, and delta loci. Of note was the discovery that half of the IGK variable (V) genes were located downstream of, and inverted with respect to, the rest of the locus. Analysis of the germline sequences of all the AR V genes identified greater conservation between dog and human than mouse with either. This work brings our understanding of the genetic diversity and expression of AR in dogs to the same completeness as that of mice and men, making it the third species to have all AR loci comprehensively and accurately annotated. The large number of germline sequences serves as a reference for future studies, and has allowed statistically powerful conclusions to be drawn on the pressures that have shaped these loci.

Investigation of the association between serum protein concentrations and concurrent chronic kidney disease in hyperthyroid cats.

Our objective was to identify if changes in serum protein concentrations occur in hyperthyroidism and to assess their association with the development of azotaemia following treatment. Initially non-azotaemic hyperthyroid cats and healthy older cats were included. Serum concentrations of protein fractions were determined by agarose gel electrophoresis and compared between; hyperthyroid and control cats, initially non-azotaemic hyperthyroid cats which developed azotaemia in a 4month follow up period (masked-azotaemic) and those which remained non-azotaemic, and hyperthyroid cats before and at the time of restoration of euthyroidism. Data are presented as median [25th, 75th percentiles]. Hyperthyroid cats (n=56) had higher serum α2 globulin concentrations (12.5 [10.9, 13.1] g/L vs. 9.8 [3.0, 11.4] g/L; P<0.001) and lower serum γ globulin concentrations (11.4 [9.1, 13.3] g/L vs. 14.0 [12.4, 16.8] g/L; P=0.001) than control cats (n=26). Following treatment, serum total globulin concentration increased (from 38.6 [35.4, 42.8] g/L to 42.3 [39.0, 45.7] g/L; P<0.001), serum α2 globulin concentration decreased (from 12.5 [10.9, 13.9] g/L to 11.5 [10.1, 12.6] g/L; P<0.001) and serum γ globulin concentration increased (from 11.4 [9.0, 13.3] g/L to 14.0 [12.4, 16.8] g/L; P<0.001). Serum concentrations of total globulin or globulin fractions were not significantly different between masked-azotaemic and non azotaemic groups. In conclusion, hyperthyroidism is associated with altered serum concentrations of the α2 and γ globulin fractions, however these changes were not associated with the development of azotaemic chronic kidney disease following treatment.

Validation of an electrophoretic method to detect albuminuria in cats.

Objectives The aims of this study were to validate a semi-automated high-resolution electrophoretic technique to quantify urinary albumin in healthy and diseased cats, and to evaluate its diagnostic performance in cases of proteinuria and renal diseases. Methods Urine samples were collected from 88 cats (healthy; chronic kidney disease [CKD]; lower urinary tract disease [LUTD]; non-urinary tract diseases [OTHER]). Urine samples were routinely analysed and high-resolution electrophoresis (HRE) was performed. Within-assay and between-assay variability, linearity, accuracy, recovery and the lowest detectable and quantifiable bands were calculated. Receiver operating curve (ROC) analysis was also performed. Results All coefficients of variation were <10%, percentage recovery was between 97% and 109% with a high linearity (r = 0.99). HRE allowed the visualisation of a faint band of albumin and a diffused band between alpha and beta zones in healthy cats, while profiles from diseased cats were variable. Albumin (mg/dl) and urine albumin:creatinine ratio (UAC) were significantly ( P <0.05) different between healthy and diseased cats. After ROC analysis, UAC values of 0.035 and 0.074 had a high sensitivity and high specificity, respectively, to classify proteinuria and identify borderline proteinuric cats. Moreover, a UAC of 0.017 had a high sensitivity in distinguishing between healthy and diseased cats. However, UAC was not able to distinguish between renal (CKD) and non-renal diseases (LUTD/OTHER), probably owing to the pathophysiology of CKD in cats, which is characterised by low-grade proteinuria and less glomerular involvement than in dogs. Conclusions and relevance HRE is an accurate and precise method that could be used to measure albuminuria in cats. UAC was useful to correctly classify proteinuria and to discriminate between healthy and diseased cats. HRE might also provide additional information on urine proteins with a profile of all proteins (albumin and globulins) to aid clinicians in the diagnosis of diseases characterised by proteinuria.

Effect of prewarming EDTA blood samples to 37°C on platelet count measured by Sysmex XT-2000iV in dogs, cats, and horses.

BACKGROUND: Pseudothrombocytopenia secondary to platelet clumping is a common cause of preanalytic error for platelet counts in dogs, cats, and horses. In human beings, it is suggested that prewarming blood samples to 37°C prior to hematology analysis will reduce platelet clumping. OBJECTIVES: The purpose of the study was to evaluate the effect of prewarming EDTA blood samples to 37°C on measured platelet counts and other hematologic variables. METHODS: The EDTA blood samples from dogs, cats and horses submitted to the clinical pathology laboratory at the University of Cambridge were included. Complete blood cell counts performed using a Sysmex XT-2000iV hematology analyzer were done on samples at room temperature (approximately 22°C) and following warming of the samples to 37°C in a water bath. The Wilcoxon signed rank test was used to compare hematologic variables, including platelet count, before and after sample warming to 37°C. Data are presented as median (25(th) , 75(th) percentile) increase. RESULTS: Blood samples from 39 dogs, 19 cats, and 10 horses were included. Sample warming to 37°C resulted in a statistically significant increase in platelet counts in dogs (11 [-2, 30] ×10(9) /L), cats (36 [14, 84] ×10(9) /L), and horses (42 [31, 79] ×10(9) /L). Sample warming did not significantly affect other hematologic variables. CONCLUSIONS: Prewarming EDTA blood samples to 37°C prior to hematologic analysis increased platelet counts overall in canine, feline, and equine blood, but did not abrogate platelet clumping and pseudothrombocytopenia fully in some cases. Furthermore, true pseudothrombocytopenia was not confirmed in these animals.

In Vitro Influence of Mycophenolic Acid on Selected Parameters of Stimulated Peripheral Canine Lymphocytes.

Mycophenolic acid (MPA) is an active metabolite of mycophenolate mofetil, a new immunosuppressive drug effective in the treatment of canine autoimmune diseases. The impact of MPA on immunity is ambiguous and its influence on the canine immune system is unknown. The aim of the study was to determine markers of changes in stimulated peripheral canine lymphocytes after treatment with MPA in vitro. Twenty nine healthy dogs were studied. Phenotypic and functional analysis of lymphocytes was performed on peripheral blood mononuclear cells cultured with mitogens and different MPA concentrations- 1 µM (10(-3) mol/m(3)), 10 µM or 100 µM. Apoptotic cells were detected by Annexin V and 7-aminoactinomycin D (7-AAD). The expression of antigens (CD3, CD4, CD8, CD21, CD25, forkhead box P3 [FoxP3] and proliferating cell nuclear antigen [PCNA]) was assessed with monoclonal antibodies. The proliferation indices were analyzed in carboxyfluorescein diacetate succinimidyl ester (CFSE)-labeled cells. All analyses were performed using flow cytometry. The influence of MPA on apoptosis was dependent on the mechanism of cell activation and MPA concentration. MPA caused a decrease in the expression of lymphocyte surface antigens, CD3, CD8 and CD25. Its impact on the expression of CD4 and CD21 was negligible. Its negative influence on the expression of FoxP3 was dependent on cell stimulation. MPA inhibited lymphocyte proliferation. In conclusion, MPA inhibited the activity of stimulated canine lymphocytes by blocking lymphocyte activation and proliferation. The influence of MPA on the development of immune tolerance-expansion of Treg cells and lymphocyte apoptosis-was ambiguous and was dependent on the mechanism of cellular activation. The concentration that MPA reaches in the blood may lead to inhibition of the functions of the canine immune system. The applied panel of markers can be used for evaluation of the effects of immunosuppressive compounds in the dog.

Validation of an automated enzyme immunoassay for the measurement of serum total thyroxine in cats.

BACKGROUND: Hyperthyroidism is common in older cats, which necessitates frequent screening of serum total thyroxine (TT4) concentrations. Fast, cheap, and reliable methods to measure TT4 in cats are needed. OBJECTIVES: The purpose of the study was the validation of a human TT4 enzyme immunoassay (EIA) for use with feline serum, and derivation of a TT4 reference interval (RI) for cats aged 9 years and older. METHODS: Assay precision, reproducibility, and linearity were evaluated. Interference by hemolysis was also assessed. Method comparison studies between the human EIA and a previously validated radioimmunoassay (RIA) and chemiluminescent-enzyme immunoassay (CEIA) were performed. Healthy cats (> 9 years) were recruited from 3 UK first opinion practices. RESULTS: The human TT4 EIA demonstrated good precision and reproducibility, and adequate linearity. Hemolysis did not significantly alter measured TT4 concentrations until HGB > 8 g/L. Method comparison revealed proportional and constant errors between EIA and RIA/CEIA. The TT4 RI for cats (> 9 years) was calculated as 7.1-45.1 nmol/L (n = 49). CONCLUSIONS: The human TT4 EIA was successfully validated for use with feline serum and offers a rapid, cheap, and reliable method for determination of serum TT4 concentrations in cats.

Over-expression of Plk4 induces centrosome amplification, loss of primary cilia and associated tissue hyperplasia in the mouse.

To address the long-known relationship between supernumerary centrosomes and cancer, we have generated a transgenic mouse that permits inducible expression of the master regulator of centriole duplication, Polo-like-kinase-4 (Plk4). Over-expression of Plk4 from this transgene advances the onset of tumour formation that occurs in the absence of the tumour suppressor p53. Plk4 over-expression also leads to hyperproliferation of cells in the pancreas and skin that is enhanced in a p53 null background. Pancreatic islets become enlarged following Plk4 over-expression as a result of equal expansion of α- and ß-cells, which exhibit centrosome amplification. Mice overexpressing Plk4 develop grey hair due to a loss of differentiated melanocytes and bald patches of skin associated with a thickening of the epidermis. This reflects an increase in proliferating cells expressing keratin 5 in the basal epidermal layer and the expansion of these cells into suprabasal layers. Such cells also express keratin 6, a marker for hyperplasia. This is paralleled by a decreased expression of later differentiation markers, involucrin, filaggrin and loricrin. Proliferating cells showed an increase in centrosome number and a loss of primary cilia, events that were mirrored in primary cultures of keratinocytes established from these animals. We discuss how repeated duplication of centrioles appears to prevent the formation of basal bodies leading to loss of primary cilia, disruption of signalling and thereby aberrant differentiation of cells within the epidermis. The absence of p53 permits cells with increased centrosomes to continue dividing, thus setting up a neoplastic state of error prone mitoses, a prerequisite for cancer development.

Gamma delta T-cell large granular lymphocyte lymphoma in a dog.

A 2-year and 6-month-old female neutered Labrador Retriever with Horner syndrome, megaesophagus, and a mediastinal mass was referred to the Queen Mother Hospital for Animals of the Royal Veterinary College. A large granular lymphocyte (LGL) lymphoma was diagnosed on cytology; flow cytometric analysis revealed a γδ T-cell phenotype (CD3+, CD5+, CD45+, TCRγδ+, CD4-, CD8-, CD34-, CD21-). Chemotherapy was started with a combination of lomustine, vincristine, procarbazine, and prednisolone, followed by bleyomicin. Euthanasia was elected by the owners, due to progressive deterioration and lack of quality of life, 28 days after diagnosis. This is the first cytologic and immunophenotypic characterization of a canine γδ T-cell lymphoma with LGL morphology and probably of mediastinal origin. The role of chemotherapy in delaying the disease progression remains unknown.

Inhibition of UDP-glucosylceramide synthase in mice prevents Gaucher disease-associated B-cell malignancy.

Clonal B-cell proliferation is a frequent manifestation of Gaucher disease - a sphingolipidosis associated with a high risk of multiple myeloma and non-Hodgkin lymphoma. Gaucher disease is caused by genetic deficiency of acid ß-glucosidase, the natural substrates of which (ß-d-glucosylceramide and ß-d-glucosylsphingosine) accumulate, principally in macrophages. Mice with inducible deficiency of ß-glucosidase [Gba(tm1Karl/tm1Karl)Tg(MX1-cre)1Cgn/0] serve as an authentic model of human Gaucher disease; we have recently reported clonal B-cell proliferation accompanied by monoclonal serum paraproteins and cognate tumours in these animals. To explore the relationship between B-cell malignancy and the biochemical defect, we treated Gaucher mice with eliglustat tartrate (GENZ 112638), a potent and selective inhibitor of the first committed step in glycosphingolipid biosynthesis. Twenty-two Gaucher mice received 300 mg/kg of GENZ 112638 daily for 3-10 months from 6 weeks of age. Plasma concentrations of ß-d-glucosylceramide and the unacylated glycosphingolipid, ß-d-glucosylsphingosine, declined. After administration of GENZ 112638 to Gaucher mice for 3-10 months, serum paraproteins were not detected and there was a striking reduction in the malignant lymphoproliferation: neither lymphomas nor plasmacytomas were found in animals that had received the investigational agent. In contrast, 14 out of 60 Gaucher mice without GENZ 112638 treatment developed these tumours; monoclonal paraproteins were detected in plasma from 18 of the 44 age-matched mice with Gaucher disease that had not received GENZ 112638. Long-term inhibition of glycosphingolipid biosynthesis suppresses the development of spontaneous B-cell lymphoma and myeloma in Gaucher mice.

Thromboelastography in healthy dairy cows.

Thromboelastography is a whole blood-based coagulation assay that can be used to investigate hypocoagulability and hypercoagulability, as seen with thromboembolic diseases and disseminated intravascular coagulation. Numerous coagulopathies due to different causes are reported in cows. The objective was to establish reference intervals for thromboelastography using the TEG 5000 (Haemonetics GmbH, Munich, Germany) with citrated whole blood samples and kaolin activation in dairy cows and to investigate possible thromboelastographic changes between cows in different lactation periods. An additional objective was to test the stability of samples for up to 100h. Sixty blood samples from healthy Holstein-Friesian cows were examined. The samples were allocated to 3 different lactation groups (≤30 d postcalving, 31-99 d postcalving, ≥100 d postcalving). Thromboelastography was performed by using the TEG 5000 analyzer with citrated whole blood samples with kaolin activation. The calculated reference intervals were as follows: reaction time=2.2 to 6.2min, coagulation time=0.8 to 2.0min, angle α=58.2 to 81.8°, maximum amplitude=64.3 to 89.2mm, and clot rigidity=9.2 to 41.2 dyn/cm(2). The 3 different lactation groups showed no significant differences in TEG parameters. No significant difference was seen in samples stored for up to 48h at room temperature, which indicates that delays in processing samples, such as those arising during transit, are not an issue.

Use of Six Sigma Worksheets for assessment of internal and external failure costs associated with candidate quality control rules for an ADVIA 120 hematology analyzer.

BACKGROUND: Quality control (QC) validation is an essential tool in total quality management of a veterinary clinical pathology laboratory. Cost-analysis can be a valuable technique to help identify an appropriate QC procedure for the laboratory, although this has never been reported in veterinary medicine. OBJECTIVE: The aim of this study was to determine the applicability of the Six Sigma Quality Cost Worksheets in the evaluation of possible candidate QC rules identified by QC validation. METHODS: Three months of internal QC records were analyzed. EZ Rules 3 software was used to evaluate candidate QC procedures, and the costs associated with the application of different QC rules were calculated using the Six Sigma Quality Cost Worksheets. The costs associated with the current and the candidate QC rules were compared, and the amount of cost savings was calculated. RESULTS: There was a significant saving when the candidate 1-2.5s, n = 3 rule was applied instead of the currently utilized 1-2s, n = 3 rule. The savings were 75% per year (£ 8232.5) based on re-evaluating all of the patient samples in addition to the controls, and 72% per year (£ 822.4) based on re-analyzing only the control materials. The savings were also shown to change accordingly with the number of samples analyzed and with the number of daily QC procedures performed. CONCLUSIONS: These calculations demonstrated the importance of the selection of an appropriate QC procedure, and the usefulness of the Six Sigma Costs Worksheet in determining the most cost-effective rule(s) when several candidate rules are identified by QC validation.