Clinical bleeding diathesis, laboratory haemostatic aberrations and survival in dogs infected with Angiostrongylus vasorum: 180 cases (2005-2019).

OBJECTIVES: Bleeding diathesis is a complication in dogs infected with Angiostrongylus vasorum. This retrospective study investigated clinical and laboratory haemostatic differences in A. vasorum-positive dogs with and without signs of bleeding and impact of bleeding on survival. MATERIALS AND METHODS: Demographics, type of clinical bleeding, haematocrit and a range of haemostatic tests, including thromboelastography and derived velocity curves were retrospectively registered from A. vasorum-positive dogs. All parameters were compared between dogs with and without signs of bleeding using univariable analyses. Binomial and multinomial regression models were applied to examine specific indicators in the bleeding dogs. P-values were false discovery rate adjusted, and adjusted P<0.05 was considered significant. RESULTS: One hundred and eighty dogs entered the study, including 65 dogs (36.1%) presenting with bleeding diathesis. Different types of cutaneous and mucosal bleeding were the most common clinical findings. Twenty dogs presented with neurological signs associated with intracranial and intra-spinal bleeding. One hundred and thirty-seven dogs had haematological and/or haemostatic laboratory analyses performed. Haematocrit, platelet count, thromboelastographic angle, maximum amplitude, global clot strength, maximum rate of thrombin generation and total thrombin generation were decreased, while prothrombin time was prolonged in bleeding dogs. Survival rate of bleeding dogs was lower at hospital discharge (76.9%) and 1 month after diagnosis (66.0%) than in dogs without signs of bleeding (94.8% and 90.1% at discharge and at 1 month, respectively). CLINICAL SIGNIFICANCE: Several haemostatic aberrations were detected in A. vasorum-positive dogs with bleeding diathesis. Bleeding was identified as an important negative prognostic indicator in A. vasorum-positive dogs.

Association between anticoagulant therapy, exacerbations and mortality in a Danish cohort of patients with Chronic Obstructive Pulmonary Disease.

OBJECTIVES: Pulmonary Embolism has been frequently reported in Acute Exacerbations of Chronic Obstructive Pulmonary Disease (AE-COPD). The study aimed to determine whether COPD patients who receive anticoagulant (AC) therapy have a reduced risk of hospitalization due to AE-COPD and death. METHODS: This nationwide population-based study was based on data from the Danish Register of COPD (DrCOPD), which contains complete data on COPD outpatients between 1st January 2010 and 31st December 2018. National registers were used to obtain information regarding comorbidities and vital status. Propensity-score matching and Cox proportional hazards models were used to assess AE-COPD and death after one year. RESULTS: The study cohort consisted of 58,067 patients with COPD. Of these, 5194 patients were on AC therapy. The population was matched 1:1 based on clinical confounders and AC therapy, resulting in two groups of 5180 patients. We found no association between AC therapy and AE-COPD or all-cause mortality in the propensity-score matched population (HR 1.03, 95% CI 0.96-1.10, p = 0.37). These findings were confirmed in a competing risk analysis. In the sensitivity analysis, we performed an adjusted analysis of the complete cohort and found a slightly increased risk of AE-COPD or death in patients treated with AC therapy. This study found a low incidence of pulmonary embolisms and deep venous thrombosis in both groups. CONCLUSIONS: AC therapy was not associated with the risk of hospitalization due to AE-COPD or all-cause mortality.

Retrospective review of 27 European cases of fatal elephant endotheliotropic herpesvirus-haemorrhagic disease reveals evidence of disseminated intravascular coagulation.

Elephant endotheliotropic herpesvirus haemorrhagic disease (EEHV-HD) is widely acknowledged as the most common cause of mortality in young Asian elephants (Elephas maximus) in captivity. The objective of the current study was to perform a blinded, retrospective pathology review of European EEHV-HD fatalities, constituting the largest systematic assessment of EEHV-HD pathology to date. Findings between viral genotypes were compared with the aim to investigate if disseminated intravascular coagulation (DIC) could be substantiated as a significant complicating factor, thereby increasing the understanding of disease pathophysiology. Immunohistochemical staining confirmed endothelial cell (EC) damage and the presence of EC intranuclear inclusion bodies, demonstrating a direct viral cytopathic effect. Microthrombi were observed in 63% of cases in several organs, including lungs, which, together with widespread haemorrhage and thrombocytopenia reported in EEHV-HD case reports, supports the presence of overt DIC as a serious haemostatic complication of active EEHV infection. Death was attributed to widespread vascular damage with multi-organ dysfunction, including severe acute myocardial haemorrhage and subsequent cardiac failure. Systemic inflammation observed in the absence of bacterial infection may be caused by cytokine release syndrome. Findings reinforce the necessity to investigate cytokine responses and haemostatic status during symptomatic and asymptomatic EEHV viraemia, to potentially support the use of anti-inflammatory treatment in conjunction with anti-viral therapy and cardiovascular support.

Sensory and motor axonal excitability testing in early diabetic neuropathy.

OBJECTIVE: The aim of the present study was to gain insight into the pathophysiology of diabetic polyneuropathy (DPN) and examine the diagnostic value of sensory and motor axonal excitability testing. METHODS: One hundred and eleven type 2 diabetics with and without DPN (disease duration: 6.36 ±â€¯0.25 years) and 60 controls were included. All participants received a thorough clinical examination including Michigan Neuropathy Screening Instrument (MNSI) score, nerve conduction studies (NCS), and sensory and motor excitability tests. Patients were compared by the likelihood of neuropathy presence, ranging from no DPN (17), possible/probable DPN (46) to NCS-confirmed DPN (48). RESULTS: Motor excitability tests showed differences in rheobase and depolarizing threshold electrotonus measures between NCS-confirmed DPN group and controls but no changes in hyperpolarising threshold electrotonus or recovery cycle parameters. Sensory excitability showed even less changes despite pronounced sensory NCS abnormalities. There were only weak correlations between the above motor excitability parameters and clinical scores. CONCLUSIONS: Changes in excitability in the examined patient group were subtle, perhaps because of the relatively short disease duration. SIGNIFICANCE: Less pronounced excitability changes than NCS suggest that axonal excitability testing is not of diagnostic value for early DPN and does not provide information on the mechanisms.

Diagnosis and prevalence of diabetic polyneuropathy: a cross-sectional study of Danish patients with type 2 diabetes.

BACKGROUND AND PURPOSE: Diabetic polyneuropathy (DPN) is a common complication of diabetes. Using the Toronto criteria for diabetic polyneuropathy and the grading system for neuropathic pain, the performance of neuropathy scales and questionnaires were assessed by comparing them to a clinical gold standard diagnosis of DPN and painful DPN in a cohort of patients with recently diagnosed type 2 diabetes. METHODS: A questionnaire on neuropathy and pain was sent to a cohort of 5514 Danish type 2 diabetes patients. A sample of 389 patients underwent a detailed clinical examination and completed neuropathy questionnaires and scales. RESULTS: Of the 389 patients with a median diabetes duration of 5.9 years, 126 had definite DPN (including 53 with painful DPN), 88 had probable DPN and 53 had possible DPN. There were 49 patients with other causes of polyneuropathy, neuropathy symptoms or pain, 10 with subclinical DPN and 63 without DPN. The sensitivity of the Michigan Neuropathy Screening Instrument questionnaire to detect DPN was 25.7% and the specificity 84.6%. The sensitivity of the Toronto Clinical Neuropathy Scoring System, including questionnaire and clinical examination, was 62.9% and the specificity was 74.6%. CONCLUSIONS: Diabetic polyneuropathy affects approximately one in five Danish patients with recently diagnosed type 2 diabetes but neuropathic pain is not as common as previously reported. Neuropathy scales with clinical examination perform better compared with questionnaires alone, but better scales are needed for future epidemiological studies.

MScanFit motor unit number estimation and muscle velocity recovery cycle recordings in diabetic polyneuropathy.

OBJECTIVE: Motor Unit Number Estimation (MUNE) methods may be valuable in tracking motor unit loss in diabetic polyneuropathy (DPN). Muscle Velocity Recovery Cycles (MVRCs) provide information about muscle membrane properties. This study aimed to examine the utility of the MScanFit MUNE in detecting motor unit loss and to test whether the MVRCs could improve understanding of DPN pathophysiology. METHODS: Seventy-nine type-2 diabetic patients were compared to 32 control subjects. All participants were examined with MScanFit MUNE and MVRCs in anterior tibial muscle. Lower limb nerve conduction studies (NCS) in peroneal, tibial and sural nerves were applied to diagnose large fiber neuropathy. RESULTS: NCS confirmed DPN for 47 patients (DPN + ), with 32 not showing DPN (DPN-). MScanFit showed significantly decreased MUNE values and increased motor unit sizes, when comparing DPN + patients with controls (MUNE = 71.3 ± 4.7 vs 122.7 ± 3.8), and also when comparing DPN- patients (MUNE = 103.2 ± 5.1) with controls. MVRCs did not differ between groups. CONCLUSIONS: MScanFit is more sensitive in showing motor unit loss than NCS in type-2 diabetic patients, whereas MVRCs do not provide additional information. SIGNIFICANCE: The MScanFit results suggest that motor changes are seen as early as sensory, and the role of axonal membrane properties in DPN pathophysiology should be revisited.

Toward better estimates of the real-time individual amino acid requirements of growing-finishing pigs showing deviations from their typical feeding patterns.

Pigs exposed to stressors might change their daily typical feeding intake pattern. The objective of this study was to develop a method for the early identification of deviations from an individual pig's typical feeding patterns. In addition, a general approach was proposed to model feed intake and real-time individual nutrient requirements for pigs with atypical feeding patterns. First, a dynamic linear model (DLM) was proposed to model the typical daily feed intake (DFI) and daily gain (DG) patterns of pigs. Individual DFI and DG dynamics are described by a univariate DLM in conjunction with Kalman filtering. A standardized tabular cumulative sum (CUMSUM) control chart was applied to the forecast errors generated by DLM to activate an alarm when a pig showed deviations from its typical feeding patterns. The relative feed intake (RFI) during a challenge period was calculated. For that, the forecasted individual pig DFI is expressed as its highest DFI relative to the intake during pre-challenge period. Finally, the DLM and RFI approaches were integrated into the actual precision-feeding model (original model) to estimate real-time individual nutrient requirements for pigs with atypical feeding patterns. This general approach was evaluated with data from two studies (130 pigs, at 35.25 ± 3.9 kg of initial BW) that investigated during 84 days the effect of precision-feeding systems for growing-finishing pigs. The proposed general approach to estimating real-time individual nutrient requirements (updated model) was evaluated by comparing its estimates with those generated by the original model. For 11 individuals out of 130, the DLM did not fit the observed data well in a specific period, resulting in an increase in the sum of standardized forecast errors and in the number of time steps that the model needed to adapt to the new patterns. This poor fit can be identified by the increase in the CUMSUM with a consequent alarm generated. The results of this study show that the updated model made it possible to reduce intra-individual variation for the estimated lysine requirements in comparison with the original model, especially for individuals with atypical feeding patterns. In conclusion, the DLM in conjunction with CUMSUM could be used as a tool for the online monitoring of DFI for growing-finishing pigs. Moreover, the proposed general approach allows the estimation of real-time amino acid requirements and accounts for the reduced feed intake and growth potential of pigs with atypical feeding patterns.

Body composition and organ development of intra-uterine growth restricted pigs at weaning.

Sow litter sizes have increased, subjecting more small piglets to intra-uterine growth restriction (IUGR). Research on the development and growth of IUGR pigs is limited. The objective of this study was to compare the body composition and organ development of IUGR pigs at weaning, and to estimate their growth performance from birth to 30 kg. A total of 142 IUGR and 142 normal piglets were classified at birth based on their head morphology. At weaning, 20 IUGR and 20 normal piglets were collected, a whole-body dual-energy x-ray absorption scan was performed, and the piglets were euthanized for organ measurements. Body weight (BW) was measured weekly from birth to 30 kg, rectal temperature and whole-blood glucose levels were measured weekly from birth to weaning, and blood samples were collected at days 7, 14 and 21 for IGF-1 analysis. Results showed that IUGR pigs have a similar percentage of adipose tissue (P > 0.05) compared to normal pigs at 24 days of age. Organs were smaller (P < 0.001) in IUGR pigs than in normal pigs, whereas brain, liver, lungs and adrenal glands were relatively larger (P < 0.05) in relation to the BW of IUGR pigs. Average birth weight (BiW) of normal pigs was greater (P < 0.001) compared with IUGR pigs (1.38 v. 0.75 kg), and the average daily gain (ADG) of IUGR pigs was reduced from day 0 to 14, day 0 to 28 (weaning) and from weaning to 30 kg compared to normal pigs. From birth to weaning at day 28, IUGR piglets had a 72.9 g/day greater fractional ADG (FADG) in relation to their BiW (P < 0.05), but FADG did not differ (P > 0.05) from weaning to 30 kg. Rectal temperature of IUGR piglets was greater (P < 0.05) on day 7 compared with normal piglets, and, even though blood glucose levels were decreased (P < 0.001) in IUGR piglets at day 0, neither glucose nor IGF-1 concentrations differed (P > 0.05) between IUGR and normal piglets. In conclusion, IUGR piglets exhibited some relatively larger organs at weaning compared to normal pigs, but body composition was similar between IUGR and normal pigs. In addition, IUGR pigs had a reduced ADG from birth to 30 kg, and, although they exhibited a greater FADG during nursing, IUGR pigs still require six additional days to reach a BW of 30 kg in comparison to normal pigs.

Probing optical resonances of silicon nanostructures using tunable-excitation Raman spectroscopy.

Optical materials with a high refractive index enable effective manipulation of light at the nanoscale through strong light confinement. However, the optical near field, which is mainly confined inside such high-index nanostructures, is difficult to probe with existing measurement techniques. Here, we exploit the connection between Raman scattering and the stored electric energy to detect resonance-induced near-field enhancements in silicon nanostructures. We introduce a Raman setup with a wavelength-tunable laser, which allows us to tune the Raman excitation wavelength and thereby identify Fabry-Pérot and Mie type resonances in silicon thin films and nanodisk arrays, respectively. We measure the optical near-field enhancement by comparing the Raman response on and off resonance. Our results show that tunable-excitation Raman spectroscopy can be used as a complimentary far-field technique to reflection measurements for nanoscale characterization and quality control. As proof-of-principle for the latter, we demonstrate that Raman spectroscopy captures fabrication imperfections in the silicon nanodisk arrays, enabling an all-optical quality control of metasurfaces.

Detection of early motor involvement in diabetic polyneuropathy using a novel MUNE method - MScanFit MUNE.

OBJECTIVE: Detection of motor involvement in diabetic polyneuropathy (DPN) by nerve conduction studies (NCS) does not occur until there is substantial loss of motor units, because collateral reinnervation maintains compound muscle action potential (CMAP) amplitude. Motor unit number estimation (MUNE) methods may therefore be more sensitive. This study was undertaken to test whether the novel method, MScanFit MUNE (MScan) can detect motor involvement in DPN despite normal NCS. METHODS: Fifty-two type-2 diabetic patients and 38 healthy controls were included. The median nerve was examined in all participants using standard NCS and a detailed CMAP scan, used for MScan. Additional lower extremity NCS in patients were used for DPN diagnosis. RESULTS: Of 52 diabetic patients, 21 had NCS-defined DPN while lower extremity NCS were normal in 31 patients. MScan motor unit number and size showed higher sensitivity and incidence of abnormality than motor NCS parameters, and a similar sensitivity to sensory NCS. CONCLUSIONS: MScan is able to detect motor axonal damage at times when collateral reinnervation limits NCS changes. SIGNIFICANCE: MScan is a sensitive method to detect motor involvement in DPN, which our data suggests is present as early as sensory.

Coagulation biomarkers and prediction of venous thromboembolism and survival in small cell lung cancer: A sub-study of RASTEN - A randomized trial with low molecular weight heparin.

Coagulation activation and venous thromboembolism (VTE) are hallmarks of cancer; however, there is an unmet need of improved biomarkers for individualized anticoagulant treatment. The present sub-study of the RASTEN trial was designed to explore the role of coagulation biomarkers in predicting VTE risk and outcome in a homogenous cancer patient population. RASTEN is a multicenter, randomized phase-3 trial investigating the survival effect of low molecular weight heparin enoxaparin when added to standard treatment in newly diagnosed small cell lung cancer (SCLC) patients. Plasma collected at baseline, during treatment, and at follow-up was used in this ad hoc sub-study (N = 242). Systemic coagulation was assessed using four assays reflecting various facets of the coagulation system: Total tissue factor (TF); extracellular vesicle associated TF (EV-TF); procoagulant phospholipids (PPL); and thrombin generation (TG). We found small variations of biomarker levels between baseline, during treatment and at follow-up, and appeared independent on low molecular weight heparin treatment. Overall, none of the measured biomarkers at any time-point did significantly associate with VTE incidence, although increased total TF at baseline showed significant association in control patients not receiving low molecular weight heparin (P = 0.03). Increased TG-Peak was significantly associated with decreased overall survival (OS; P = 0.03), especially in patients with extensive disease. Low baseline EV-TF predicted a worse survival in the low molecular weight heparin as compared with the control group (HR 1.42; 95% CI 1.04-1.95; P = 0.03; P for interaction = 0.12). We conclude that the value of the analyzed coagulation biomarkers for the prediction of VTE risk was very limited in SCLC patients. The associations between TG-Peak and EV-TF with patient survival and response to low molecular weight heparin therapy, respectively, warrant further studies on the role of coagulation activation in SCLC aggressiveness.

A comparative analysis of heterogeneity in commercially available recombinant factor VIII products.

INTRODUCTION: Advances in analytical technologies enable investigation of possible correlations between molecular structure, aggregation and subvisible particle content. Regulatory agencies place increasing attention on potential risks associated with protein aggregates in the micron range in biological therapeutics. AIM: Assess the heterogeneity, high-molecular-weight protein (HMWP) species, subvisible particle content and posttranslational modifications in six commercially available recombinant FVIII (rFVIII) products. METHODS: Three B-domain-deleted (BDD) or B-domain truncated rFVIII products (turoctocog alfa, simoctocog alfa and moroctocog alfa) and three full-length rFVIII products (octocog alfa FS and two octocog alfa) were analysed. HMWP content, amount of micron range subvisible particles, tyrosine-1680 sulphation and N-glycan analysis were investigated. RESULTS: The B-domain-modified products had more protein size homogeneity vs the full-length products. Size exclusion-high-performance liquid chromatography data indicated no association between B-domain structure and aggregate content or size of the products tested. The rFVIII products showed large variation in subvisible particle concentration, with turoctocog alfa and simoctocog alfa having the lowest numbers (1000-1600 and 1800-2400 particles/100 IU, respectively). Turoctocog alfa and simoctocog alfa displayed the most complete tyrosine sulphation (>99.5%). CONCLUSION: Overall, there was no association between molecular structure (full-length B-domain, BDD or truncated) and subvisible particle or HMWP content. Dissimilarities may be related to production and product handling differences. In this study, turoctocog alfa, such as simoctocog alfa, had one of the lowest levels of subvisible particles and HMWP content, and high protein size homogeneity.

The utility of motor unit number estimation methods versus quantitative motor unit potential analysis in diagnosis of ALS.

OBJECTIVE: To compare the diagnostic utility of motor unit number estimation (MUNE) methods to motor unit potential (MUP) analysis in amyotrophic lateral sclerosis (ALS). METHODS: Twenty-five patients (1 definite, 11 probable, 9 possible ALS and 4 progressive muscular atrophy) and 22 healthy controls were prospectively included. Quantitative MUP analysis and three MUNE methods; Multiple Point Stimulation MUNE (MPS), Motor Unit Number Index (MUNIX) and MScanFit MUNE (MScan) were done in abductor pollicis brevis muscle. The sensitivities were compared by McNemar chi-square test. MUNE, MUP and revised ALS Functional Rating Scale (ALSFRS-R) parameters were correlated by regression analysis. RESULTS: The sensitivities of MPS (76%) and MScan (68%) were higher than MUP duration (36%) and amplitude (40%) in detecting motor unit loss (p < 0.05). MUNE methods increased the categorical probability from possible to probable ALS in 4 patients (16%). There was only significant correlation between ALSFRS-R and MScan (r = 0.443, p = 0.027) among the electrophysiological tests. MUNE methods did not correlate to MUP parameters. CONCLUSIONS: MUNE methods are more sensitive in showing abnormality than MUP analysis. SIGNIFICANCE: MUNE methods, in particular MScan, may have the potential to be implemented in the clinical practice for diagnosis and follow-up of neuromuscular disorders particularly ALS.

Liposome-encapsulated chemotherapy: Current evidence for its use in companion animals.

Cytotoxic drugs encapsulated into liposomes were originally designed to increase the anticancer response, while minimizing off-target adverse effects. The first liposomal chemotherapeutic drug was approved for use in humans more than 20 years ago, and the first publication regarding its use in a canine cancer patient was published shortly thereafter. Regardless, no general application for liposomal cytotoxic drugs has been established in veterinary oncology till now. Due to the popularity of canines as experimental models for pharmacokinetic analyses and toxicity studies, multiple publications exist describing various liposomal drugs in healthy dogs. Also, some evidence for its use in veterinary cancer patients exists, especially in canine lymphoma, canine splenic hemangiosarcoma and feline soft tissue sarcoma, however, the results have not been overwhelming. Reasons for this may be related to inherent issues with the enhanced permeability and retention effect, the tumour phenomenon which liposomal drugs exploit. This effect seems very heterogeneously distributed in the tumour. Also, it is potentially not as ubiquitously occurring as once thought, and it may prove important to select patients for liposomal therapy on an individual, non-histology-oriented, basis. Concurrently, new developments with active-release modified liposomes in experimental models and humans will likely be relevant for veterinary patients as well, and holds the potential to improve the therapeutic response. It, however, does not resolve the other challenges that liposomal chemotherapy faces, and more work still needs to be done to determine which veterinary patients may benefit the most from liposomal chemotherapy.

Effects of manipulating tetanic calcium on the curvature of the force-velocity relationship in isolated rat soleus muscles.

AIM: In dynamically contracting muscles, increased curvature of the force-velocity relationship contributes to the loss of power during fatigue. It has been proposed that fatigue-induced reduction in [Ca++ ]i causes this increased curvature. However, earlier studies on single fibres have been conducted at low temperatures. Here, we investigated the hypothesis that curvature is increased by reductions in tetanic [Ca++ ]i in isolated skeletal muscle at near-physiological temperatures. METHODS: Rat soleus muscles were stimulated at 60 Hz in standard Krebs-Ringer buffer, and contraction force and velocity were measured. Tetanic [Ca++ ]i was in some experiments either lowered by addition of 10 µmol/L dantrolene or use of submaximal stimulation (30 Hz) or increased by addition of 2 mmol/L caffeine. Force-velocity curves were constructed by fitting shortening velocity at different loading forces to the Hill equation. Curvature was determined as the ratio a/F0 with increased curvature reflecting decreased a/F0 . RESULTS: Compared to control levels, lowering tetanic [Ca++ ]i with dantrolene or reduced stimulation frequency decreased the curvature slightly as judged from increase in a/F0 of 13 ± 1% (P = < .001) and 20 ± 2% (P = < .001) respectively. In contrast, increasing tetanic [Ca++ ]i with caffeine increased the curvature (a/F0 decreased by 17 ± 1%; P = < .001). CONCLUSION: Contrary to our hypothesis, interventions that reduced tetanic [Ca++ ]i caused a decrease in curvature, while increasing tetanic [Ca++ ]i increased the curvature. These results reject a simple causal relation between [Ca++ ]i and curvature of the force-velocity relation during fatigue.

Analyses of body weight patterns in growing pigs: a new view on body weight in pigs for frequent monitoring.

Frequent BW monitoring of growing pigs can be useful for identifying production (e.g. feeding), health and welfare problems. However, in order to construct a tool which will properly recognize abnormalities in pigs' growth a precise description of the growth process should be used. In this study we proposed a new model of pig growth accounting for daily fluctuations in BW. Body weight measurements of 1710 pigs (865 gilts and 843 barrows) originating from five consecutive batches from a Danish commercial farm were collected. Pigs were inserted into a large pen (maximum capacity=400) between November 2014 and September 2015. On average, each pig was observed for 42 days and weighed 3.6 times a day when passing from the resting to feeding area. Altogether, 243,160 BW measurements were recorded. A multilevel model of pig growth was constructed and fitted to available data. The BW of pigs was modeled as a quadratic function of time. A diurnal pattern was incorporated into the model by a cosine wave with known length (24 h). The model included pig effect which was defined as a random autoregressive process with exponential correlation. Variance of within-pigs error was assumed to increase with time. Because only five batches were observed, it was not possible to obtain the random effect for batch. However, in order to account for the batch effect the model included interactions between batch and fixed parameters: intercept, time, square value of time and cosine wave. The gender effect was not significant and was removed from the final model. For all batches, morning and afternoon peaks in the frequency of visits to the feeding area could be distinguished. According to results, pigs were lighter in the morning and heavier in the evening (minimum BW was reached around 1000 h and maximum around 2200 h). However, the exact time of obtaining maximum and minimum BW during the day differed between batches. Pigs had access to natural light and, therefore, existing differences could be explained by varying daylight level during observations periods. Because the diurnal amplitude for pig growth varied between batches from 0.9 to 1.4 kg, BW monitoring tools based on frequent measurements should account for diurnal variation in BW of pigs. This proposed description of growth will be built into a monitoring tool (a dynamic linear model) and applied to farm data in future studies.

Diagnosis of canine pulmonary thromboembolism by computed tomography and mathematical modelling using haemostatic and inflammatory variables.

There is no evidence-based diagnostic approach for diagnosis of pulmonary thromboembolism (PTE) in dogs. Many dogs with diseases that predispose to thrombosis are hypercoagulable when assessed with thromboelastography (TEG), but no direct link has been established. The aims of this study were: (1) to investigate if diseased dogs with PTE, diagnosed by computed tomography pulmonary angiography (CTPA), had evidence of hypercoagulability by TEG; (2) to characterise haemostatic and inflammatory changes in dogs with PTE; (3) to construct models for prediction of PTE based on combinations of haemostatic and inflammatory variables; and (4) to evaluate the performance of D-dimer measurement for prediction of PTE. Twenty-five dogs were included in this prospective observational study (PTE: n=6; non-PTE: n=19). Clot strength G values did not differ between the PTE and non-PTE groups in tissue factor (TF) or kaolin-activated TEG analyses. Haemostatic and inflammatory variables did not differ between the two groups. Linear discriminant analysis generated a model for prediction of PTE with a sensitivity and specificity of 100% when TF results were used as TEG data, and a model with sensitivity of 83% and specificity of 100% when kaolin results were used as TEG data. Receiver operating characteristic analysis of D-dimer levels showed that a value of >0.3mg/L yielded a sensitivity of 100% and a specificity of 71.4%. In conclusion, the study supports CTPA as method for diagnosing canine PTE, but shows that TEG alone cannot identify dogs with PTE. Models for prediction of PTE were generated, but require further validation.

Complete sequence-based pathway analysis by differential on-chip DNA and RNA extraction from a single cell.

We demonstrate on-chip, differential DNA and RNA extraction from a single cell using a microfluidic chip and a two-stage lysis protocol. This method enables direct use of the whole extract, without additional washing steps, reducing sample loss. Using this method, the tumor driving pathway in individual cells from a colorectal cancer cell line was determined by applying a Bayesian computational pathway model to sequences obtained from the RNA fraction of a single cell and, the mutations driving the pathway were determined by analyzing sequences obtained from the DNA fraction of the same single cell. This combined functional and mutational pathway assessment of a single cell could be of significant value for dissecting cellular heterogeneity in tumors and analyzing single circulating tumor cells.

Circulating let-7g is down-regulated in Bernese Mountain dogs with disseminated histiocytic sarcoma and carcinomas - a prospective study.

Cancer is a prevalent cause of mortality in Bernese mountain dogs (BMDs). Circulating microRNAs (miRNAs) are found in blood and have been identified as promising biomarkers in various neoplastic diseases in humans. In the current study, the expression profile of different types of miRNAs was investigated in healthy BMDs and BMDs with cancer. Seven healthy and six non-treated BMDs with cancer [four with disseminated histiocytic sarcomas (DHS)] were enrolled in this study. Clinical evaluations including physical examination, blood analysis, urinalysis and diagnostic imaging were performed on all dogs. Twenty-four different miRNAs were profiled from RNA isolated from whole blood preserved in PAXgene® tubes using quantitative real-time PCR (qPCR). The miRNA let-7g was significantly down-regulated in dogs with cancer (P = 0.002) and dogs with DHS (P = 0.011) compared with healthy controls. This miRNA is a known tumour suppressor and further analyses are warranted to assess its value as a non-invasive biomarker for early detection of different types of cancer in BMDs.

Use of nutritional supplements by Danish elite athletes and fitness customers.

The nutritional supplement (NS) industry is one of the fastest growing in the world, and NS use in Denmark is among the highest in Europe. However, the exact use in elite athletes and fitness customers targeted for doping control is unknown. Information from 634 doping control forms obtained in 2014 was evaluated (elite athletes: n = 361; fitness customers: n = 273). The majority of female (92.6%) and male (85.0%) elite athletes and female (100.0%) and male (94.0%) fitness customers declared using one or more NS. The use of non-ergogenic NS was more prevalent in women than in men and in younger (15-34 years) compared with older (35-49 years) subjects, but it was less prevalent in intermittent compared with endurance and power/strength sports. Additionally, fitness customers who tested positive for doping also reported using more NS than subjects testing negative, indicating an association between NS and doping abuse. The present results demonstrate a very high prevalence of NS usage in both elite athletes and fitness customers. This highlights the importance of a strong national regulation of NS to avoid contamination of NS with doping substances.