“Arte et Labore”—A Blackburn Rovers fan's legacy in human complex trait genetics

Through his own research contributions on the modelling and genetic analysis of quantitative traits and through his former students and postdocs, Robin Thompson has indirectly left a major legacy in human genetics. In this short note, we highlight examples of the long‐lasting relevance and impact of Robin's work in human genetics. A lone early study of marker‐assisted selection developed many of the tools and approaches later exploited (often after reinvention) by the human genetics community in GWAS studies and for prediction. Furthermore, a particularly clear example of the pervasive impact of Robin's work is that REML has become the default method to estimate variance components and that genetic predictions exploiting linkage disequilibrium in the population are starting to become used in precision medicine applications.     

A “taxonomic affidavit”: Why it is needed?

Imprecise and faulty taxonomic identification of the biological objects of many ecological and experimental studies renders these studies irreproducible. Incomplete identification to the species level and excessive use of vernacular species names are additional problems. Good science must be able to be falsified. I recommend that publications and granting agencies use and mention the names of the zoologists or botanists who identified the species. Voucher specimens should be marked and deposited in scientific museums for future checking.     

24‐Hour Kinetics of Cardiac Troponin‐T Using a “High‐Sensitivity” Assay in Thoroughbred Chuckwagon Racing Geldings after Race and Associated Clinical Sampling Guidelines

Background

A “high‐sensitivity” cardiac troponin‐T (hscTnT) assay recently has been validated for use in horses and is a specific biomarker of myocardial damage. Postexercise release kinetics of cTnT utilizing the hscTnT assay have yet to be established in horses.

Objectives

To determine: (1) cTnT release kinetics in racing Thoroughbreds after a high‐intensity 5/8th mile Chuckwagon race; (2) the effects of age on pre‐ and postrace cTnT concentrations; and (3) sampling guidelines for clinicians evaluating horses presenting after exercise.

Animals

Samples were obtained from 38 Thoroughbred geldings aged 5–16 years before racing and immediately, 2, 3, 4, 6, 12, and 24 hour postrace.

Methods

Prospective, observational study with convenience sampling. A fifth‐generation hscTnT assay was used for plasma sample analysis, and concentrations were compared at all time‐points. Correlations were determined between cTnT concentrations and age. Biochemistry analysis was performed to assess rhabdomyolysis, renal failure, and exercise‐induced dehydration.

Results

All horses with measureable cTnT concentrations had significant postexercise increases in cTnT with a median peak (8.0 ng/L) at 3‐hour postrace. All horses had peak postexercise cTnT concentrations 2‐ to 6‐hour postrace ≤ the 99th percentile upper reference limit of 23.2 ng/L, after which all cTnT concentrations decreased until returning to baseline by 12–24 hours. There was no correlation over time between cTnT concentrations and age.

Conclusions and Clinical Importance

In racing Thoroughbreds completing short‐duration, high‐intensity Chuckwagon races, cTnT concentrations are expected to be increased 2‐ to 6‐hour postrace and to decrease by 12–24 hours while remaining ≤23.2 ng/L throughout. This study contributes to establishing guidelines for clinical use of the hscTnT assay in exercising horses.     

Unusual “erythroid loops” in canine blood smears after viper‐bite envenomation

Abstract: Northern Italy is a habitat for many species of viper; Vipera aspis is responsible for most reported bites of humans and animals. Five dogs of different breeds and ages were presented by their owners to a veterinary clinic in northern Italy between September 2004 and August 2007 with a history of being bitten by a viper within the past 2 hours. On physical examination, all of the dogs were depressed, had pale mucous membranes, and had a painful area consistent with a recent viper bite wound on the nose (2), distal front leg (2), or lip (1). Hemoglobinemia and hemoglobinuria were observed in plasma and urine from all dogs. CBCs were done at the time of presentation, daily for 4 days, and 10 days after presentation in all dogs with the exception of one dog that died after 2 days. All dogs had an acute decrease in HCT within 24 hours of presentation; all dogs had neutrophilia, 3/5 had a mildly toxic left shift, and 4/5 had thrombocytopenia. On Diff‐Quik‐stained blood smears, moderate numbers of echinocytes, spherocytes, and erythrocyte ghosts were observed. In addition, moderate numbers of unusual erythrocyte membrane‐like structures (“erythroid loops”) were observed. The loops were annular in shape and sometimes disrupted, appearing as thin pale blue bands. Erythrocyte morphologic abnormalities decreased by day 3 and were no longer observed on day 10. The unique appearance of the erythroid loops together with evidence for intravascular hemolysis and other erythrocyte morphologic changes suggest they may be a consequence of erythrocyte lysis. Echinocytes, spherocytes, and erythrocyte ghosts are known to result from the action of phospholipase in viper venom; however, erythroid loops have not been reported previously and their exact mechanism of formation is unknown.     

Evaluation of Pax5 expression and comparison with BLA.36 and CD79αcy in feline non‐Hodgkin lymphoma

Paired box gene 5 (Pax5) is a widely used B‐cell marker for human and canine non‐Hodgkin's lymphoma (nHL); however, in the literature there is only one case report using Pax5 in a cat B‐cell lymphoma. The purposes of this study were to investigate the expression and detection of B‐cell specific activator protein (BSAP) using a monoclonal anti‐Pax5 antibody in feline nHL (FnHL) tissue samples to evaluate its diagnostic relevance as a B‐cell marker. A total of 45 FnHL samples in 45 cats were evaluated. B‐cell lymphoma was the most common immunophenotype (51.1%) for all the samples and T‐cell the most common immunophenotype (64.3%) for the gastrointestinal (GI) form. Pax5 stained 82.6% of all B‐cell lymphomas and no expression was found in any of the T‐cell lymphomas. Anti‐Pax5 antibody staining in FnHL is similar to that reported in human and canine counterparts and may offer an excellent B‐cell marker in cats.     

Selecting for changes in average “parity curve” pattern of litter size in Large White pigs

This study aimed to analyse genetic background of variation in reproductive performance between parities of a sow and to investigate selection strategies to change the “parity curve”. Total number born (TNB) recorded in Large White sows was provided by Topigs Norsvin. Analysis with basic (BM) and random regression (RRM) models was done in ASReml 4.1. The BM included only a fixed “parity curve”, while RRM included 3rd order polynomials for additive genetic and permanent sow effects. Parameters from RRM were used in simulations in SelAction 2.1. Based on Akaike information criterion, RRM was a better model for TNB data. Genetic variance and heritability estimates of TNB from BM and RRM were increasing with parity from parity 2. Genetically, parity 1 is the most different from parities 7 to 10, whereas most similar to parities 2 and 3. This indicates presence of genetic variation to change the “parity curve”. Based on simulations, the selection to increase litter size in parity 1 only increases TNB in all parities, but does not change the observed shape of “parity curve”, whereas selection for increased TNB in parity 1 and reduced TNB in parity 5 decreases differences between parities, but also reduces overall TNB in all parities. Changing the “parity curve” will be difficult as the genetic and phenotypic relationships between the parities are hard to overcome even when selecting for one parity.     

Using “Uterine Index” to diagnose embryonic death in mares

Embryonic death plays an important role in infertility in mares, and most of embryonic mortality occurs at an early stage (before day 40) of pregnancy. Previous investigations focused on B-mode sonographic images to determine abnormal pregnancy; however, no study has provided a quantitative and objective method to diagnose embryonic death. Here we developed a new index, “Uterine Index,” using color Doppler sonography to describe alterations of uterine blood flow in physiological and pathological gestations during early gestation. Six Trotter mares were used in this study. The pregnancies were interrupted with a prostaglandin F2α (PGF_2α) analog (tiaprost) at days 25, 30, and 40. Three gestations were investigated in each stage. After the PGF_2α administration, these mares were examined every 8 hours until the embryonic vesicles were not visible on ultrasound. In the control group, three pregnancies were examined daily from days 12 to 60. The embryo size was measured in B-mode sonography, and the blood flow in uterine arteries was examined in Doppler ultrasound. The resistance index (RI) and uterine index (UI) were calculated. In the control group, the RI values of uterine artery were lower in pregnant side compared with that of the non-pregnant side from day 15 on, and the difference became greater as the pregnancy advanced (P < .05). The UI was close to 0 before week 3 and increased to approximately 10 after week 4. In the PGF_2α-treated group, the UI values reduced from pregnancy level to approximately 0. As early as 32 hours before embryonic death, we observed a significant difference of UI values between control and treated mares. On the contrary, the embryo size of treated mares was not different from that of normal mares. Through the analysis of all recorded UI values, we suggest that the UI values 10 and 5 are good values for distinguishing normal pregnancy (UI >10) from embryonic death (UI < 5). If the UI values are located between 10 and 5, a reexamination at intervals of 1 to 3 days is recommended. Our findings demonstrate that the UI is a useful method to diagnose and predict embryonic death in mares by providing reliable and objective information.     

A PROPOSED RADIOGRAPHIC CLASSIFICATION SCHEME FOR CONGENITAL THORACIC VERTEBRAL MALFORMATIONS IN BRACHYCEPHALIC “SCREW‐TAILED” DOG BREEDS

Congenital vertebral malformations are common in brachycephalic “screw‐tailed” dog breeds such as French bulldogs, English bulldogs, Boston terriers, and pugs. The aim of this retrospective study was to determine whether a radiographic classification scheme developed for use in humans would be feasible for use in these dog breeds. Inclusion criteria were hospital admission between September 2009 and April 2013, neurologic examination findings available, diagnostic quality lateral and ventro‐dorsal digital radiographs of the thoracic vertebral column, and at least one congenital vertebral malformation. Radiographs were retrieved and interpreted by two observers who were unaware of neurologic status. Vertebral malformations were classified based on a classification scheme modified from a previous human study and a consensus of both observers. Twenty‐eight dogs met inclusion criteria (12 with neurologic deficits, 16 with no neurologic deficits). Congenital vertebral malformations affected 85/362 (23.5%) of thoracic vertebrae. Vertebral body formation defects were the most common (butterfly vertebrae 6.6%, ventral wedge‐shaped vertebrae 5.5%, dorsal hemivertebrae 0.8%, and dorso‐lateral hemivertebrae 0.5%). No lateral hemivertebrae or lateral wedge‐shaped vertebrae were identified. The T7 vertebra was the most commonly affected (11/28 dogs), followed by T8 (8/28 dogs) and T12 (8/28 dogs). The number and type of vertebral malformations differed between groups (P = 0.01). Based on MRI, dorsal, and dorso‐lateral hemivertebrae were the cause of spinal cord compression in 5/12 (41.6%) of dogs with neurologic deficits. Findings indicated that a modified human radiographic classification system of vertebral malformations is feasible for use in future studies of brachycephalic “screw‐tailed” dogs.     

A retrospective analysis of radiation therapy for the treatment of feline vaccine‐associated sarcoma*

We retrospectively evaluated predictive prognostic factors in 73 cats with vaccine‐associated sarcoma given postsurgical curative (n = 46, most with clean margins) or coarse fractionated radiotherapy (n = 27, most with either macroscopic disease or dirty margins). The former animals displayed a median survival of 43 months and a median progression free interval (PFI) of 37 months, the latter reached a median survival of 24 months and a median PFI of 10 months. In cats undergoing coarse fractionated therapy, factors predictive of a better outcome included lack of visible mass (n = 10) as opposed to macroscopic disease (n = 17, survival: 30 versus 7 months, P = 0.025; PFI: 20 versus 4 months, P = 0.01), adjuvant chemotherapy for gross disease (n = 5/17, survival: 29 versus 5 months, P = 0.04) and a smaller number of surgeries preceding radiation therapy (coeff = 0.41, P = 0.03). The Ki67 index was not predictive for survival. We concluded that postsurgical curative and coarse fractionated radiotherapy are effective legitimate options for managing vaccine‐associated sarcomas.     

Measurements of hypoxia ([18F]‐FMISO, [18F]‐EF5) with positron emission tomography (PET) and perfusion using PET ([15O]‐H2O) and power Doppler ultrasonography in feline fibrosarcomas*

The aim of this study was to evaluate if hypoxia in feline fibrosarcomas can be detected. This was done using positron emission tomography (PET), two hypoxia tracers and polarographic pO₂ measurements. Of the seven cats included, five received [¹⁸F]‐fluoromisonidazole and two 2‐(2‐nitro‐1H‐imidazol‐1‐yl)‐N‐(2,2,3,3,3‐pentafluoropropyl) acetamide. Perfusion was evaluated with [¹⁵O]‐H₂O (n = 4) and with contrast‐enhanced power Doppler ultrasonography (n = 5). Hypoxia was detected in three cats. Polarographic pO₂ measurements did not confirm PET results. In the ultrasonographic evaluation, low vascularity and low perfusion were seen with a peripheral vascular pattern and no perfusion in the centre of the tumour. This was in contrast to the [¹⁵O]‐H₂O scans, where central perfusion of the tumour was also found. In conclusion, it appears that hypoxia exists in this tumour type. The presence of tumour necrosis and heterogeneous hypoxia patterns in these tumours may explain the found discrepancies between the applied techniques.     

Decreased blood cell counts were not observed in cattle living in the “difficult‐to‐return zone” of the Fukushima nuclear accident

White blood cells, especially lymphocytes, are susceptible to radiation exposure. In the present study, red blood cell, total white blood cell, and lymphocyte counts were repeatedly measured in cattle living on three farms located in the “difficult‐to‐return zone” of the Fukushima nuclear accident, and compared with two control groups from unaffected areas. Blood cell counts differed significantly between the two control groups, although almost all the values fell within the normal range. The blood cell counts of the cattle in the “difficult‐to‐return zone” varied across sampling times even on the same farms, being sometimes higher or lower than either of the two control groups. However, neither a statistically significant decrease in blood cell counts nor an increase in the rate of cattle with extremely low blood cell counts was observed overall. The estimated cumulative exposure dose for the cattle on the most contaminated farm was within a range of 500–1000 mSv, exceeding the threshold for the lymphopenia. Because of the low dose rate on these farms, potential radiation damages would have been repaired and have not accumulated enough to cause deterministic effects.     

IMAGING DIAGNOSIS —ANTEMORTEM DETECTION OF OLIGODENDROGLIOMA “CEREBROSPINAL FLUID DROP METASTASES” IN A DOG BY SERIAL MAGNETIC RESONANCE IMAGING

An English Bulldog underwent radiation therapy of an intracranial, left lateral ventricle mass. Following resolution of the primary mass, an intraventricular fourth ventricle lesion developed. Subsequently, multiple lesions developed from the cervical central canal and leptomeninges. Serial magnetic resonance imaging documented the propagation of lesions along the cerebrospinal fluid (CSF) pathways, known as “CSF drop metastasis.” Histopathology confirmed multifocal intraventricular and leptomeningeal oligodendroglioma. Oligodendroglioma should be included in the differential diagnosis for an intraventricular tumor exhibiting apparent CSF drop metastasis.