Quantification of circulating tumour cells over time in dogs with appendicular osteosarcoma

Enumeration of circulating tumour cells (CTC) has shown promise for prognostication and guidance of therapeutic decisions in human cancers. The objective of this study was to enumerate CTC over time in dogs with naturally occurring osteosarcoma (OSA), and to determine correlation with patient outcome. Twenty-six dogs with OSA and no evidence of metastatic disease at the time of amputation were enrolled. Dogs were assessed for lung metastases and CTC prior to and following amputation, and at each chemotherapy visit. Twenty-one dogs completed the study. Nineteen dogs were euthanized and two were alive and free of metastases. Overall survival time ranged from 88 to 1058 days (median survival time (MST) 374 days). Increased serum alkaline phosphatase activity, advanced age, and higher body weight were significantly associated with lower MST. Dogs with OSA had a mean of 356 (0 to 4443) CTC/10⁶ leukocytes. In 12 of 15 dogs that developed radiographic evidence of metastasis, a pre-metastatic CTC spike was retrospectively detectable on average 36.5 (1–100 days) days prior to metastasis and was associated with significantly shorter MST (301 ± 64 vs. 626 ± 55 days; p = .0107). In a multivariable analysis, dogs with a CTC spike were 10× more likely to die compared with those without. These results suggest that a spike in CTC frequency precedes detection of metastasis in dogs with OSA and is associated with shorter survival. More frequent enumeration of CTC in a larger cohort of dogs with OSA may be warranted.     

Dermatosparaxis in two Limousin calves

Background

An unusual presentation of skin disease was identified in two related neonatal Pedigree Limousin calves presented to University Veterinary Hospital, University College Dublin, following detailed post mortem examination a diagnosis of dermatosparaxis was made. Dermatosparaxis in animals or Ehlers Danlos Syndrome, which is the analogous condition seen in humans, is a connective tissue disorder characterised by extreme skin fragility. To the authors’ knowledge this is the first report of such a diagnosis in the Limousin breed and the features of this lethal phenotype were severe in comparison to previous reports of the condition.

Case presentation

Two calves, which were full siblings, a pedigree Limousin bull (Calf A) and pedigree Limousin heifer (Calf B) were examined clinically after presenting collapsed since birth, both had grossly abnormal skin with multiple skin fissures visible and both calves were subsequently euthanised. Both calves underwent gross post mortem examination, after which histological samples were reviewed and electron microscopical examination of selected skin samples was carried out. Histological features of dysplastic dermal collagen were identified. The diagnosis of dermatosparaxis in the Limousin breed was confirmed. Genetic testing was conducted to determine if the current cases had the same mutation as has previously been described in Belgian Blue cattle. Some common parentage was traced but genetic testing did not show a similar mutation to that previously described in cattle. The specific genetic cause in this case is unknown.

Conclusions

This is the first report of dermatosparaxis in the Limousin and the presentation of the dermatosparaxis phenotype has some noteworthy features thus further genetic testing is required to pinpoint the causative mutation or other genetic defect. Given the popularity of the breed and the lethal nature of the phenotype in this case it is important to raise awareness of the condition.

     

Evaluation of coagulation via thromboelastography in healthy horses administered dexamethasone

Dexamethasone was administered to healthy horses daily for 7 days. Blood samples were collected at 3 time points from both treatment and non-treatment groups, and analyzed via thromboelastography (TEG). There were no significant differences in TEG parameters between treated and untreated horses, or within treatment groups over time.     

Anatomical and ultrasonographic study of the femoral nerve within the iliopsoas muscle in beagle dogs and cats

ObjectivesAn ultrasound (US)-guided ventral suprainguinal approach to block the femoral nerve (FN) within the iliopsoas muscle (IPM) has recently been described in dogs. The goal of the present study was to provide the operator with additional information to locate the FN within the IPM in dogs and cats using US.Study designThe study was carried out in three phases: a dissection of the FN (phase 1); an in vivoUS-assisted nerve study (phase 2), and an anatomical cross-sectional study (phase 3).AnimalsNine healthy adult beagle dogs and nine healthy adult cats.MethodsDissections were performed to investigate the anatomical characteristics of the FN and its related structures in one dog and one cat. Ultrasound scans of the left and right FN were performed in eight dogs and eight cats. The FN diameter and the distance between the FN and the external iliac artery (EIA) in US images and in anatomical cryosections were measured.ResultsThe median FN diameter did not differ significantly between cats and dogs (1.1 mm versus 1.0 mm) or between the two techniques (US versus anatomical cross-sectional study) (1.1 mm versus 1.1 mm in dogs; 1.0 mm versus 1.1 mm in cats). The US and anatomical measurements of the median distances between the FN and EIA differed significantly between dogs and cats (8.2 mm versus 5.8 mm by US; 5.7 mm versus 4.8 mm in the anatomical study).Conclusions and clinical relevanceThe distance between the EIA and FN is reproducible in beagle dogs and cats and can be used in locating the FN within the IPM.     

Individual seedling mortality as a function of size, growth and competition in naturally regenerated beech seedlings

Young forest stands that are naturally regenerated are characterizedby a large number of seedlings and high mortality rates. Goodprediction of seedling mortality is therefore necessary to modelthe dynamics of young stands. We used data obtained in a naturalbeech (Fagus sylvatica L.) regeneration in north-eastern France,to fit logistic models to predict seedling mortality and toanalyse how seedling size, seedling growth and local competitionrelate to seedling mortality. Good relationships were obtainedwith recent seedling diameter growth, and with a combinationof local competition and initial seedling size. However, therelationships between the probability of mortality and recentgrowth appeared to be unstable and to vary substantially withlocal competition and among measurement years, precluding theuse of these relationships in regeneration models.     

DNA sequence polymorphisms and their application to bread wheat quality

Single-nucleotide polymorphisms (SNPs) constitute an abundant source of DNA polymorphisms, which have been successfully used to identify loci that are associated with a particular phenotype. Additionally, such markers could be efficiently used in combination with doubled haploid technology to improve the efficiency of breeding programmes. Information on such markers in plants is still scarce. For bread wheat, SNP data are restricted to a few genes. This can be explained by the hexaploidy of Triticum aestivum which makes SNP discovery difficult. We developed a novel method for SNP discovery in bread wheat. The strategy is based on the development of highly specific PCR-primers, which were used to sequence 27 lines. SNPs were discovered from sequence alignment data. Some SNPs were identified by mass spectrometry in a collection of 113 lines, which were both evaluated for agronomic traits and genotyped at 42 neutral microsatellite loci. Traits investigated include: protein content, the quantity of high-molecular-weight glutenins and that of the GluBx subunit. The 42 markers were used to infer population structure, which was included in linear models for association studies. The results of this preliminary study showed 89 SNPs in approximately 20 kbp, i.e., one SNP every 223 bp on average. Six SNPs were genotyped: three were located along the sequence of Glu-B1-1, while three non-synonymous SNPs were located along the sequence of the B homoeologous gene coding for SPA (Storage Protein Activator). The SNPs from Glu-B1-1 had a significant effect on the studied variables, whereas those of SPA had no effect. Such results might indicate that some haplotypes for Glu-B1-1 are linked to higher protein content, through an increased amount of high-molecular-weight glutenins, especially the GluBx subunit.