Feline panleukopenia virus in cerebral neurons of young and adult cats.

BACKGROUND: Perinatal infections with feline panleukopenia virus (FPV) have long been known to be associated with cerebellar hypoplasia in kittens due to productive infection of dividing neuroblasts. FPV, like other parvoviruses, requires dividing cells to replicate which explains the usual tropism of the virus for the digestive tract, lymphoid tissues and bone marrow in older animals. RESULTS: In this study, the necropsy and histopathological analyses of a series of 28 cats which died from parvovirus infection in 2013 were performed. Infections were confirmed by real time PCR and immunohistochemistry in several organs. Strikingly, while none of these cats showed cerebellar atrophy or cerebellar positive immunostaining, some of them, including one adult, showed a bright positive immunostaining for viral antigens in cerebral neurons (diencephalon). Furthermore, infected neurons were negative by immunostaining for p27(Kip1), a cell cycle regulatory protein, while neighboring, uninfected, neurons were positive, suggesting a possible re-entry of infected neurons into the mitotic cycle. Next-Generation Sequencing and PCR analyses showed that the virus infecting cat brains was FPV and presented a unique substitution in NS1 protein sequence. Given the role played by this protein in the control of cell cycle and apoptosis in other parvoviral species, it is tempting to hypothesize that a cause-to-effect between this NS1 mutation and the capacity of this FPV strain to infect neurons in adult cats might exist. CONCLUSIONS: This study provides the first evidence of infection of cerebral neurons by feline panleukopenia virus in cats, including an adult. A possible re-entry into the cell cycle by infected neurons has been observed. A mutation in the NS1 protein sequence of the FPV strain involved could be related to its unusual cellular tropism. Further research is needed to clarify this point.

Assessment of Toll-like receptor 2, 4 and 9 SNP genotypes in canine sino-nasal aspergillosis.

BACKGROUND: The exact aetiology of canine sino-nasal aspergillosis (SNA) is unknown. In man, dysfunction in innate immunity, particularly in the function of pattern recognition receptors, is implicated in the pathogenesis of inflammatory sino-nasal disease and in fungal diseases. Associations between single nucleotide polymorphisms (SNPs) in Toll-like receptors (TLRs) and these diseases have been identified. Similarly, in dogs SNPs in genes encoding TLRs may be important in the pathogenesis of SNA. The aims of the present study were (1) to identify the presence of non-synonymous SNPs in the coding regions of the TLR2, 4 and 9 genes in dogs suffering from SNA, and (2) to investigate the SNP genotypes in dogs with SNA compared with a control population. RESULTS: Direct sequencing of nine dogs of various breeds with SNA revealed two non-synonymous SNPs in the coding region of TLR2, eight in TLR4 and four in TLR9. These non-synonymous SNPs were further evaluated in a case-control study of affected Golden Retrievers, Labrador Retrievers, Rottweilers and Beaucerons. Genotyping was performed using a combination of allele-specific primers and hydrolysis probe assays in 31 dogs with SNA and 31 controls. No significant difference in minor allele frequency was identified between these groups, for all studied SNPs, in any of the four breeds. CONCLUSIONS: These findings do not support a role for non-synonymous SNPs in the TLR 2, 4 and 9 coding regions in the pathogenesis of canine SNA, but do not exclude a role for innate immunity in the pathogenesis of the disease.

Fasting triglyceride concentrations are associated with markers of lipid metabolism and glucose homeostasis in healthy, non-obese dogs in lean and overweight condition.

Serum triglyceride concentrations increase in dogs with obesity, which is typically assessed by body condition score (BCS), however little is known about changes that take place in non-obese dogs in overweight condition. Further, the associations of triglyceride levels with other markers of energy homeostasis are poorly characterised in healthy animals. The present study aimed to evaluate associations between both BCS and triglyceride concentrations with other markers of lipid and glucose metabolism in healthy, non-obese dogs, as well as to assess whether these markers change significantly in non-obese dogs with overweight as compared to their lean counterparts. Serum concentrations of cholesterol, free fatty acids, triglycerides, insulin, glucose and fructosamine were measured in 532 healthy, client-owned dogs, assigned either to 'lean' (BCS: 3-5) or 'overweight' (BCS: 6-7) categories. Generalised linear mixed models were used to assess associations between BCS categories, triglyceride concentrations and other variables, correcting for the effect of breed. Compared with lean dogs, overweight dogs had a greater serum cholesterol concentration (95% CI, 5.3-6.2 mmol/L or 205-237 mg/dL versus 5.1-5.4 mmol/L or 198-210 mg/dL, p = 0.0032), insulin concentration (95% CI, 17.5-22.1 µU/ml versus 16.7-18.0 µU/ml, p = 0.0374) and were older (95% CI, 4.0-5.3 versus 3.4-3.7 years, p = 0.0005). Triglyceride concentrations were positively associated with fructosamine (r 2 = 0.31, p = 0.0012), cholesterol (r 2 = 0.25, p < 0.0001), insulin (r 2 = 0.14, p = 0.0030) and glucose (r 2 = 0.10, p = 0.0014) concentrations, and negatively associated with free fatty acid concentrations (r 2 = 0.11, p < 0.0001). However, there was no association between triglyceride concentrations and age. In conclusion, both BCS and triglyceride concentrations were associated with other markers of glucose and lipid metabolism in non-obese healthy dogs, amongst which those with overweight showed metabolic changes as compared to their lean counterparts. Triglyceride concentrations were associated with an increase in insulin and fructosamine concentrations that might reflect an early-phase impairment in glucose tolerance which, surprisingly, was concurrent with lower basal free fatty acid concentrations.

Feeding Cyprinus carpio with infectious materials mediates cyprinid herpesvirus 3 entry through infection of pharyngeal periodontal mucosa.

Cyprinid herpesvirus 3 (CyHV-3), also known as Koi herpesvirus, is the etiological agent of a mortal disease in common and koi carp. Recently, we investigated the entry of CyHV-3 in carp using bioluminescence imaging and a CyHV-3 recombinant strain expressing luciferase (LUC). We demonstrated that the skin is the major portal of entry after inoculation of carp by immersion in water containing CyHV-3. While this model of infection mimics some natural conditions in which infection takes place, other epidemiological conditions could favour entry of virus through the digestive tract. Here, we investigated whether ingestion of infectious materials mediates CyHV-3 entry through the digestive tract. Carp were fed with materials contaminated with the CyHV-3 LUC recombinant (oral contamination) or immersed in water containing the virus (contamination by immersion). Bioluminescence imaging analyses performed at different times post-infection led to the following observations: (i) the pharyngeal periodontal mucosa is the major portal of entry after oral contamination, while the skin is the major portal of entry after contamination by immersion. (ii) Both modes of inoculation led to the spreading of the infection to the various organs tested. However, the timing and the sequence in which some of the organs turned positive were different between the two modes of inoculation. Finally, we compared the disease induced by the two inoculation modes. They led to comparable clinical signs and mortality rate. The results of the present study suggest that, based on epidemiological conditions, CyHV-3 can enter carp either by skin or periodontal pharyngeal mucosal infection.

Idiopathic chylothorax and lymphedema in 2 whippet littermates.

Idiopathic chylothorax and limb edema was diagnosed in two 2-year-old male whippet siblings. The fact that the 2 related animals developed similar clinical signs at a young age may suggest a congenital or hereditary etiology.

Clinical safety of computed tomography-guided injection of autologous muscle-derived mesenchymal stem cells in the intervertebral disc in dogs.

Background: Pre-clinical randomized controlled animal trials have been conducted to evaluate the effect of mesenchymal stem cell (MSCs) transplantation on intervertebral disc (IVD) degeneration. MSCs can be obtained from different tissues, but systematic studies concerning the effects of muscle-derived MSCs injections on canine naturally degenerated IVD are still lacking. The aim of this study is the assessment of the clinical safety of this technique and its effects on the imaging features of the lumbosacral IVD. Methods: Eight adult healthy Beagle dogs were used in this study. In the preliminary phase, viability of muscle-derived MSCs in presence of contrast medium was assessed. In the clinical assessment phase, MSCs were injected in the lumbosacral IVD by computed-tomography (CT) guidance, after the injection of contrast medium to assess the correct intradiscal needle position. Regular clinical examinations were performed and pre- and post-injections (CT) and magnetic resonance imaging (MRI) features of the IVD were assessed. Results: The percentage of viability of MSCs in the presence of contrast medium ranged from 90 to 98%. 3x106 MSCs were obtained from six dogs and injected in the IVD. No major or minor complications were reported during the procedure and no abnormalities were noticed during the clinical examinations. No statistically significant variations were noticed between the pre- and post-injections imaging features. Conclusion: This technique is clinically safe and it is not associated with any progression of the IVD degeneration, detected by CT and MRI imaging.

Efficacy of intrasinusal administration of bifonazole cream alone or in combination with enilconazole irrigation in canine sino-nasal aspergillosis: 17 cases.

This study evaluated the effect of 1% bifonazole cream in the treatment of canine sino-nasal aspergillosis (SNA). The cream was instilled through perendoscopically placed catheters into the frontal sinuses and was used either as single therapy after debridement (DC) or as adjunctive therapy after 2% enilconazole infusion (DEC). Twelve dogs were treated initially with DEC: 7 and 3 of these dogs were free of disease after 1 and 2 procedures, respectively, while 2 dogs were cured after DC was used as a second procedure. Five dogs were treated with DC only: in 3 dogs with moderate disease, cure was obtained after a single procedure while, in 2 debilitated patients, cure could not be confirmed. Topical administration of 1% bifonazole cream appears as an effective therapy in SNA, either as an adjunctive therapy to enilconazole infusion or as sole therapy in moderately affected patients.

Developmental change in number line estimation: A strategy-based perspective.

The current study investigated developmental changes in children's benchmark-based strategy use in number line estimation. Third and fifth graders solved a 0-1,000 number line estimation task in one of three conditions. In the control condition, only the origin and endpoint were specified, the midpoint condition included an additional benchmark at 50%, and the quartile condition contained three additional benchmarks at 25%, 50%, and 75%. Trial-by-trial verbal strategy reports revealed that fifth graders, in comparison to third graders, spontaneously applied quartile-based strategies more frequently while they used strategies based on the origin, midpoint, and endpoint about equally often. Finally, children's mathematics achievement was positively related to the variety and frequency of benchmark-based strategies as well as their number line estimation performance. We conclude that developmental changes in number line estimation performance can at least partially be attributed by a refinement in children's benchmark-based strategy use. (PsycINFO Database Record (c) 2019 APA, all rights reserved).

Cardiovascular biomarkers in dogs with systemic inflammatory response syndrome.

OBJECTIVE: To measure plasma N-terminal fragments of pro-B-type natriuretic peptides (NT-proBNP) and cardiac troponin T (cTnT) concentration in hospitalized dogs and relate these markers to underlying conditions and evaluate their potential as prognostic markers in dogs with systemic inflammatory response syndrome (SIRS). DESIGN: Prospective, observational, clinical study. SETTING: Emergency department of a university teaching hospital. ANIMALS: Sixty-nine dogs with SIRS examined in the emergency department were prospectively studied. Patient age ranged from 5 months to 15 years, and weight ranged from 5.5 to 75 kg. MEASUREMENTS AND MAIN RESULTS: Blood samples were obtained at presentation, during hospitalization until discharge or death, and at a "control" visit (T1m) at least 1 month after hospital discharge. NT-proBNP was assayed with a commercially available canine ELISA, while cTnT was measured with an automated immunoassay previously used in dogs. A correlation procedure, mixed procedure on a linear model, and a logistic procedure were performed. Forty-four patients survived, 19 of which had control visits. cTnT concentrations were significantly higher than T0 and T1m at T12, T24, and T72. In 28 dogs, cTnT was detected during hospitalization, but cTnT was not detected in any dog at the control visits. Higher concentrations of cTnT were negatively associated with survival, irrespective of disease category. NT-proBNP concentrations were significantly higher than T0, T6, T12, and T1m at T24, T72, and T120, but were not associated with survival. CONCLUSIONS: NT-proBNP and cTnT increased significantly in dogs with SIRS, regardless of the underlying disease process. Nonsurvivors displayed significantly higher cTnT concentrations during hospitalization.

Whole blood and tissue fungal DNA quantification in the diagnosis of canine sino-nasal aspergillosis.

Various combinations of tests are used to confirm the diagnosis of canine sino-nasal aspergillosis (SNA) because false-positive and false-negative results can occur with each test. Therefore, the aim of this study was to evaluate whether detection of fungal DNA in blood and nasal tissue samples was of value in the clinical diagnosis of this disease. Four groups were included in the study (dogs with SNA, lymphoplasmacytic rhinitis or nasal neoplasia, and control animals). Real-time PCR assays detecting DNA from all Penicillium and Aspergillus species (PenAsp assay) or species-specific DNA from A. fumigatus, A. terreus, A. flavus and A. niger were applied to whole blood and nasal tissue samples. Results obtained by PCR were compared between the groups. Sensitivity, specificity, positive and negative predictive values (PPV and NPV) for fungal DNA detection were compared with those for alternative diagnostic procedures including histopathology, serology and fungal culture. Significantly more fungal DNA was detected by the PenAsp assay in tissue biopsies from dogs with SNA than in the three other groups. Sensitivity, specificity, PPV and NPV for this method were 1.00, 0.06, 0.32 and 1.00. A. fumigatus DNA was detected in seven tissue biopsies from dogs with SNA and in one biopsy from a dog with a nasal tumour. Sensitivity, specificity, PPV and NPV for this diagnostic test were 0.50, 0.97, 0.87 and 0.82. No significant difference was found between the groups with respect to the amount of DNA detected in blood by the PenAsp assay. Sensitivity, specificity, PPV and NPV for this method were 0.71, 0.24, 0.31 and 0.64. A. fumigatus DNA was detected in the blood of three dogs with SNA and sixteen dogs without SNA. Sensitivity, specificity, PPV and NPV for this diagnostic tool were 0.21, 0.45, 0.15 and 0.54. Detection of A. fumigatus DNA in nasal tissue had the highest specificity, PPV and NPV but sensitivity of this method was low. Detection of fungal DNA in whole blood was of no value in the diagnosis of SNA.

Comparison of bronchoalveolar lavage cytospins and smears in dogs and cats.

Differences in the cytological interpretation of bronchoalveolar lavage fluid (BALF) after cytospin preparation (CP) or manual smearing of pelleted cells preparation (MSP) were investigated in client-owned dogs and cats with inflammatory or infectious lower respiratory disease. Bronchoalveolar lavage fluid from healthy cats was also examined. With MSP, cell lysis was more frequently observed, and cellular distribution was more heterogeneous throughout the slide. When samples from healthy and diseased animals were considered together, a significantly greater percentage of neutrophils was seen on CP than on MSP slides (P<0.002). Cytospin preparations were considered of better quality in all individual comparisons. Cytospin preparation is advised in the evaluation of BALF with low total cell count. When only MSPs are evaluated, clinicians should be aware that differential neutrophil counts may underestimate the counts found on CP slides.

Benchmark-based strategy use in atypical number lines.

Previous studies have indicated that the presence of atypical end points (e.g., 1,639 and 2,897) on a number line has a negative effect on number line estimation (NLE) performance (Booth & Newton, 2012; Hurst, Leigh Monahan, Heller, & Cordes, 2014). In the present study, we investigated whether this effect could be attributed to a disruption in the ease with which benchmarks on the number line can be determined and whether this possible disruption changes with age. Hence, we asked 5th graders and adults to perform a NLE task in a typical and an atypical condition with a number line ranging from 0 to 1,000 and from 367 to 1,367, respectively. Results showed that participants' overall estimates were less accurate in the atypical condition compared to the typical condition. A similar pattern of results was observed for the estimates around the self-created benchmarks at 50% in children and at 25%, 50%, and 75% in adults. Moreover, in both age groups, we found a shift from a larger proportion of estimation patterns' being best fit by more complex power models in the typical condition toward a larger proportion of estimation patterns best described by simpler power models in the atypical condition. Of importance, we also found evidence that adults' estimation patterns in the typical condition are best described by a 4-cycle power model, hereby extending previous modeling results. Overall, our findings indicate that hindering the application of benchmark-based strategies negatively affects children's and adults' NLE performance. (PsycINFO Database Record (c) 2018 APA, all rights reserved).

Inflammatory cytokine and C-reactive protein concentrations in dogs with systemic inflammatory response syndrome.

OBJECTIVE: To evaluate C-reactive protein (CRP), interleukin 6 (IL-6), and tumor necrosis factor alpha (TNF-α) kinetics in dogs with a systemic inflammatory response syndrome (SIRS) presented to an emergency service. We hypothesized serum CRP concentrations would increase and vary during hospitalization, and would correlate with plasma IL-6 and TNF-α concentrations, vary in magnitude according to the underlying disease, and predict survival. DESIGN: Prospective, observational, clinical study. SETTING: University emergency department. ANIMALS: Sixty-nine dogs with SIRS weighing over 5 kg who could tolerate the blood sampling. INTERVENTIONS: Serum and plasma were collected (and stored at -80°C) at presentation (T0), after 6 (T6), 12 (T12), 24 (T24), and 72 (T72) hours, and at a follow-up visit at least 1 month after discharge (T1m). Underlying diseases were categorized as infection (I), neoplasia (N), trauma (T), gastric-dilation and volvulus (GDV), other gastrointestinal (GI), renal (R), and miscellaneous (M) disease. MEASUREMENTS AND MAIN RESULTS: Serum CRP concentration was measured using a canine-specific immunoturbidimetric assay. Biologically active plasma IL-6 and TNF-α concentrations were assessed using bioassays. Forty-four dogs survived, 8 died, and 17 were euthanized. Nineteen dogs had follow-up visits. At T0, serum CRP concentration was above the reference interval in 73.1% (49/67), and was within the reference interval (0-141.9 nmol/L) throughout hospitalization in only 6% (4/67). Serum CRP concentrations were significantly higher (P < 0.0001) at T0 (882.9 ± 1082.9 nmol/L) and at all time points during hospitalization (P < 0.0001) compared to T1m, with highest concentrations observed at T24 (906. 7 ± 859.0 nmol/L). At T1m, serum CRP concentrations were within the reference interval (22.9 ± 42.9 nmol/L) in 95% (18/19) of dogs. Logarithmic concentrations of serum CRP and plasma IL-6 were significantly correlated (P < 0.001, r = 0.479). None of the measured cytokines were associated with disease category or outcome. CONCLUSIONS: Serum CRP concentration is increased in dogs with SIRS, and decreases during treatment and hospitalization. Serum CRP, plasma IL-6, and plasma TNF-α concentrations cannot predict outcome in dogs with SIRS.

Interbreed variation of biomarkers of lipid and glucose metabolism in dogs.

BACKGROUND: Markers of lipid and glucose metabolism are used in both clinical practice and research. Detection of abnormal laboratory results often relies on species-specific reference intervals, but interbreed variation can also affect data interpretation. OBJECTIVES: The purpose of the present study was to compare concentrations of selected biochemical variables among different dog breeds. METHODS: We analyzed a database containing information on biochemical variables from 534 dogs belonging to nine different breeds. All dogs were confirmed to be healthy based on history, physical examination, and ancillary tests. Concentrations of glucose, fructosamine, insulin, cholesterol, triglycerides, fatty acids, and C-reactive protein were compared using the nonparametric Kruskal-Wallis and Dunn's tests. RESULTS: All variables tested showed significant interbreed differences, although all breeds remained within the previously established RIs for dogs. Fructosamine, insulin, and cholesterol showed a wide interbreed variation that could affect the interpretation of results. CONCLUSIONS: Breed is an important factor to consider when assessing energy metabolism in dogs, especially for markers like fructosamine, insulin, and cholesterol, which vary considerably among breeds.

Development and application of multiple internal reference (housekeeper) gene assays for accurate normalisation of canine gene expression studies.

Measurement of mRNA expression by real-time RT-PCR (QRT-PCR) has proven to be an important and powerful tool for the investigation of the pathogenesis of inflammatory and immune-mediated diseases in many species. This methodology has proven particularly valuable in the dog, a species for which there are currently few specific antibodies for measurement of relevant proteins. Internal control (housekeeper) mRNAs are widely used for normalisation of QRT-PCR results. The validation and use of multiple internal control mRNAs for increased accuracy of normalisation has been described for humans and rodents. The aims of this study were to develop QRT-PCR assays for 11 potential internal control mRNAs in the dog (ACTB, B(2)M, G3PDH, HMBS, HPRT1, RPL13A, RPL32, RPS18, SDHA, TBP and YWAZ) and validate their use with bone marrow, colon, duodenum, heart, kidney, liver, lung, lymph node, skeletal muscle, pancreas, spleen and stomach from seven dogs. Endoscopic biopsies of the superficial duodenal mucosa were also obtained from nine dogs suffering from chronic gastro-oesophageal disease. The most stably expressed genes varied in the tissues examined. RPL13A and RPL32 (both components of the 60S ribosomal subunit) were the most stably expressed genes in the majority of the tissues examined, whereas ACTB and B(2)M were the least stable. Distinct internal control genes were shown to be most appropriate for use in full-thickness versus superficial mucosal biopsies of the duodenum. The results of this study indicate that there are no universal control genes for gene expression studies in canine tissues. It is important to use multiple internal control genes based upon a survey of potential control genes applied to representative samples from different disease groups, culture conditions and/or time points in an experimental study.

Update on canine sinonasal aspergillosis.

Sinonasal aspergillosis is a frequent cause of nasal discharge that occurs in otherwise healthy, young to middle-aged dogs. A local immune dysfunction is suspected in affected animals, and the role of increased interleukin-10 mRNA expression in the nasal mucosa of affected dogs is currently under investigation. Despite recent advances in imaging techniques, the "gold standard" for diagnosing the disease is direct visualization of fungal plaques during endoscopy or observation of fungal elements on cytology or histopathologic examination. Treatment can be challenging; however, the use of topical enilconazole or clotrimazole through noninvasive techniques has increased the success of treatment and decreased the morbidity and duration of hospitalization.

Canine eosinophilic bronchopneumopathy.

Eosinophilic bronchopneumopathy (EBP) is a disease characterized by eosinophilic infiltration of the lung and bronchial mucosa, as demonstrated by examination of bronchoalveolar lavage fluid cytologic preparations or histologic examination of the bronchial mucosa. Although the precise cause of EBP is unknown, a hypersensitivity to aeroallergens is suspected. The diagnosis relies on typical history and clinical signs, demonstration of bronchopulmonary eosinophilia by cytology or histopathologic examination, and exclusion of known causes of lower airway eosinophilia. Most dogs display an excellent response to oral corticosteroid therapy; however, side effects of this treatment can be limiting. New therapeutic approaches are being studied, including the use of aerosol therapy, cyclosporine, or drugs interfering with T helper 2 immune response.

Benchmark-based strategies in whole number line estimation.

In this study, we used verbal protocols to identify whether adults spontaneously apply quartile-based strategies or whether they need additional external support to use these strategies when solving a 0-1,000 number line estimation (NLE) task. Participants were assigned to one of three conditions based on the number of external benchmarks provided on the number line. In the bounded condition only the origin and endpoint were indicated, the mid-point condition included an additional external benchmark at 50%, and in the quartile condition three additional external benchmarks at 25%, 50%, and 75% were specified. Firstly, participants in the bounded condition reported to spontaneously apply quartile-based strategies to calibrate their estimates. Moreover, participants frequently relied on the external benchmarks for creating internal benchmarks at the mid-point, quartiles, and even octiles of the number line. Secondly, overall estimation accuracy improved as the number of external benchmarks increased, and target numbers close to external benchmarks were estimated more accurately and with less variability. Thirdly, the use of a larger variety in benchmark-based strategies was positively related to NLE accuracy. In summary, this study provides evidence that the NLE task induces more sophisticated strategy use in participants than initially anticipated.

Evaluating the Effect of Labeled Benchmarks on Children's Number Line Estimation Performance and Strategy Use.

Some authors argue that age-related improvements in number line estimation (NLE) performance result from changes in strategy use. More specifically, children's strategy use develops from only using the origin of the number line, to using the origin and the endpoint, to eventually also relying on the midpoint of the number line. Recently, Peeters et al. (unpublished) investigated whether the provision of additional unlabeled benchmarks at 25, 50, and 75% of the number line, positively affects third and fifth graders' NLE performance and benchmark-based strategy use. It was found that only the older children benefitted from the presence of these benchmarks at the quartiles of the number line (i.e., 25 and 75%), as they made more use of these benchmarks, leading to more accurate estimates. A possible explanation for this lack of improvement in third graders might be their inability to correctly link the presented benchmarks with their corresponding numerical values. In the present study, we investigated whether labeling these benchmarks with their corresponding numerical values, would have a positive effect on younger children's NLE performance and quartile-based strategy use as well. Third and sixth graders were assigned to one of three conditions: (a) a control condition with an empty number line bounded by 0 at the origin and 1,000 at the endpoint, (b) an unlabeled condition with three additional external benchmarks without numerical labels at 25, 50, and 75% of the number line, and (c) a labeled condition in which these benchmarks were labeled with 250, 500, and 750, respectively. Results indicated that labeling the benchmarks has a positive effect on third graders' NLE performance and quartile-based strategy use, whereas sixth graders already benefited from the mere provision of unlabeled benchmarks. These findings imply that children's benchmark-based strategy use can be stimulated by adding additional externally provided benchmarks on the number line, but that, depending on children's age and familiarity with the number range, these additional external benchmarks might need to be labeled.

Quantification of mRNA encoding cytokines and chemokines in nasal biopsies from dogs with sino-nasal aspergillosis.

Canine sino-nasal aspergillosis is usually caused by Aspergillus fumigatus and is similar to human chronic erosive non-invasive fungal sinusitis. The pathogenesis of the disease is poorly understood. We investigated the nature of the local immune response mounted in canine sino-nasal aspergillosis. Quantitative RT-PCR was carried out on RNA isolated from nasal biopsies from diseased and control dogs, using specific assays designed to amplify mRNA encoding a panel of cytokines and chemokines. Canine sino-nasal aspergillosis was associated with significantly increased expression of mRNA encoding MCP-1, -2, -3 and -4, IL-8, IL-10, IL-18 and TNF-alpha relative to controls (P<0.01) but there was no difference between groups with respect to IL-4, IL-5, IL-6, IL-12, TGF-beta, and eotaxin-2 and -3. The up-regulation of proinflammatory cytokines and chemokines related to the influx of phagocytic cells might account for the localisation of this infection to the upper respiratory tract. The up-regulation of the expression of the immunomodulatory cytokine IL-10 in nasal tissue from affected dogs might be important in limiting the extent of local tissue destruction, but might also account for the fact that infected dogs are generally unable to clear this infection spontaneously.