Association of the FGF4L2 retrogene with fibrocartilaginous embolic myelopathy in dogs.

BACKGROUND: Fibrocartilaginous embolic myelopathy (FCE) is a well-documented condition in dogs although rarely reported in chondrodystrophic breeds. Genetic associations have not been defined. OBJECTIVES: Define the association of the chondrodystrophy-associated FGF4L2 retrogene with histopathologically confirmed cases of FCE. ANIMALS: Ninety-eight dogs with a histopathologic diagnosis of FCE. METHODS: Retrospective multicenter study. Dogs were genotyped for the FGF4L2 and FGF4L1 retrogenes using DNA extracted from formalin-fixed, paraffin-embedded tissue. Associations between breed, FCE and retrogene status were investigated with reference to a hospital population and known breed and general population allele frequencies. RESULTS: FGF4L2 genotype was defined in 89 FCE cases. Fibrocartilaginous embolic myelopathy was present in 22 dogs from FGF4L2-segregating breeds with allele frequencies of ≥5%; however, all dogs were wild type. Two Labrador retrievers with FCE carried FGF4L2 alleles. Frequency of the FGF4L2 allele was significantly (P < .001) and negatively associated with FCE relative to predicted hospital-population dogs. FCE was overrepresented in Boxer, Great Dane, Yorkshire Terrier, Bernese Mountain Dog, Miniature Schnauzer, Rottweiler, and Shetland Sheepdog breeds. CONCLUSIONS AND CLINICAL IMPORTANCE: Study data based on genotypically and histopathologically defined cases support the historical observation that FCE is uncommon in chondrodystrophic dog breeds. FGF4 plays an important role in angiogenesis and vascular integrity; anatomical studies comparing chondrodystrophic and non-chondrodystrophic dogs might provide insight into the pathogenesis of FCE.

Compulsory care of individuals with severe substance use disorders and alcohol- and drug-related mortality: A Swedish registry study.

Aim: This study used 17 year of Swedish registry data (2003-2019) for 25,125 adults assessed for their severity of substance use to identify the baseline factors predicting the risk of being court-ordered into compulsory care and examine the association between admission to compulsory care and mortality risks due to alcohol- or drug-related causes. Methods and materials: Addiction Severity Index (ASI) assessment data were linked to register data on demographic characteristics, compulsory care, and alcohol- and drug-related mortality. Cox regression models were used to identify baseline factors predictive of post-assessment admission to compulsory care in the 5 years post-substance use assessment. Discrete-time random-effect logistic regression models were used to examine the association between compulsory care duration and alcohol or drug-related mortality risks. Propensity score matching was used for validation. Results: The first models identified that younger age, female gender, and ASI composite scores for drug use, mental health and employment were significantly associated with the risk of placement in compulsory care for drugs other than alcohol. Female gender and ASI composite scores for alcohol, drug use and employment were significantly associated with compulsory care treatment for alcohol use. The second models showed that older individuals and men were more likely to die due to alcohol-related causes, while younger individuals and men were more likely to die due to drug-related causes. Length of stay in compulsory care institutions significantly increased the likelihood of dying due to substance use-related causes. Propensity scores analyses confirmed the results. Conclusion: In Sweden, a significant concern is the higher likelihood of women and young individuals to be court-ordered to compulsory care. Although compulsory care is often advocated as a life-saving intervention, our findings do not provide strong support for this claim. On the contrary, our findings show that admission to compulsory care is associated with a higher risk of substance use-related mortality. Factors such as compulsory care often not including any medical or psychological therapy, together with relapse and overdose after discharge, may be possible contributing factors to these findings.

Genetics of randomly bred cats support the cradle of cat domestication being in the Near East.

Cat domestication likely initiated as a symbiotic relationship between wildcats (Felis silvestris subspecies) and the peoples of developing agrarian societies in the Fertile Crescent. As humans transitioned from hunter-gatherers to farmers ~12,000 years ago, bold wildcats likely capitalized on increased prey density (i.e., rodents). Humans benefited from the cats' predation on these vermin. To refine the site(s) of cat domestication, over 1000 random-bred cats of primarily Eurasian descent were genotyped for single-nucleotide variants and short tandem repeats. The overall cat population structure suggested a single worldwide population with significant isolation by the distance of peripheral subpopulations. The cat population heterozygosity decreased as genetic distance from the proposed cat progenitor's (F.s. lybica) natural habitat increased. Domestic cat origins are focused in the eastern Mediterranean Basin, spreading to nearby islands, and southernly via the Levantine coast into the Nile Valley. Cat population diversity supports the migration patterns of humans and other symbiotic species.

Prevalence of Genetic Mutations in Horses With Muscle Disease From a Neuromuscular Disease Laboratory.

Deleterious genetic variants are an important cause of skeletal muscle disease. Immunohistochemical evaluation of muscle biopsies is standard for the diagnosis of muscle disorders. The prevalence of alleles causing hyperkalemic periodic paralysis (HYPP), malignant hyperthermia (MH), polysaccharide storage myopathy 1 (PSSM1), glycogen branching enzyme deficiency (GBED), myotonia congenita (MC), and myosin heavy chain myopathy (MYHM) in horses with muscle disease is unknown. Archived slides processed for immunohistochemical analysis from 296 horses with muscle disease were reviewed blinded and clinical information obtained. DNA isolated from stored muscle samples from these horses were genotyped for disease variants. Histological findings were classified as myopathic in 192, neurogenic in 41, and normal in 63 horses. A third of the population had alleles that explained disease which constituted 45% of the horses with confirmed histological myopathic process. Four of six muscle disease alleles were identified only in Quarter horse breeds. The allele causing PSSM1 was detected in other breeds, and MC was not detected in these samples. The My allele, associated with susceptibility for MYHM, was the most common (62%) with homozygotes (16/27) presenting a more severe phenotype compared to heterozygotes (6/33). All cases with the MH allele were fatal upon triggering by anesthesia, stress or concurrent myopathy. Both, muscle histological and genetic analyses are essential in the investigation of muscle disease, since 10% of the horses with muscle disease and normal histology had a muscle disease causing genetic variant, and 63% of histologically confirmed muscle with alterations had no known genetic variants.

The association between history of civil commitment for severe substance use and future imprisonment: A Swedish registry study.

BACKGROUND: Civil commitment for individuals with severe substance use is fairly common and a part of many treatment systems worldwide. In Sweden, individuals with severe substance use and experience with civil commitment are more likely to use higher levels of alcohol and drugs, to be younger, and be more socially marginalized compared to their counterparts. The study examined whether civil commitments for severe substance use increased the likelihood of imprisonment following the civil commitment. METHOD: Baseline ASI-data merged with national registry data on prison sentences (2007 through 2016). Cox regression was used to estimate, for a Swedish sample of 12,044 adults assessed for risky substance use, the importance of having a history of civil commitment for severe substance use, controlling for age, gender and baseline assessment of ASI composite scores in seven areas (alcohol, drugs other than alcohol, legal, mental- and physical health, family & social relationships and employment) on the likelihood of future imprisonment. RESULTS: The regression showed that being a male, those with experience of civil commitment and elevated ASI composite scores for both legal and employment were significantly associated with imprisonment post-civil commitment. Civil commitment for severe substance use showed 1.29 (HR = 1.29, 95% CI: 1.03-1.49, p < 0.001) increased likelihood of imprisonment post-civil commitment. CONCLUSION: Having been in treatment through civil commitment due to severe substance use was strongly associated with imprisonment post-civil commitment episode. This is concerning since civil commitment is supposed to mediate against the consequences of severe substance use and promote voluntary treatment participation. Those with severe substance use and a history of civil commitment are in need of a well-coordinated and integrated system of extensive aftercare services to reduce the likelihood of imprisonment.

Increased α-tocopherol metabolism in horses with equine neuroaxonal dystrophy.

BACKGROUND: Equine neuroaxonal dystrophy/equine degenerative myeloencephalopathy (eNAD/EDM) is an inherited neurodegenerative disorder associated with a vitamin E deficiency within the first year of life. Vitamin E consists of 8 isoforms metabolized by the CYP4F2 enzyme. No antemortem diagnostic test currently exists for eNAD/EDM. HYPOTHESIS/OBJECTIVES: Based on the association of α-tocopherol deficiency with the development of eNAD/EDM, we hypothesized that the rate of α-tocopherol, but not γ-tocopherol or tocotrienol metabolism, would be increased in eNAD/EDM-affected horses. ANIMALS: Vitamin E metabolism: Proof of concept (POC) study; eNAD/EDM-affected (n = 5) and control (n = 6) horses. Validation study: eNAD/EDM-affected Quarter Horses (QHs; n = 6), cervical vertebral compressive myelopathy affected (n = 6) horses and control (n = 29) horses. CYP4F2 expression and copy number: eNAD/EDM-affected (n = 12) and age- and sex-matched control (n = 11-12) horses. METHODS: The rates of α-tocopherol/tocotrienol and γ-tocopherol/tocotrienol metabolism were assessed in equine serum (POC and validation) and urine (POC only) using liquid chromatography tandem mass spectrometry (LC-MS/MS). Quantitative reverse-transcriptase PCR (qRT-PCR) and droplet digital (dd)-PCR were used to assay expression and genomic copy number of a CYP4F2 equine ortholog. RESULTS: Metabolic rate of α-tocopherol was increased in eNAD/EDM horses (POC,P < .0001; validation, P = .03), with no difference in the metabolic rate of γ-tocopherol. Horses with eNAD/EDM had increased expression of the CYP4F2 equine orthologue (P = .02) but no differences in copy number. CONCLUSIONS AND CLINICAL IMPORTANCE: Increased α-tocopherol metabolism in eNAD/EDM-affected QHs provides novel insight into alterations in vitamin E processing in eNAD/EDM and highlights the need for high-dose supplementation to prevent the clinical phenotype in genetically susceptible horses.

Correction: Weich, K., et al. Pigment Intensity in Dogs Is Associated with a Copy Number Variant Upstream of KITLG. Genes 2020, 11, 75.

The authors wish to make the following corrections to this paper [...].

Whole-genome sequencing identifies missense mutation in GRM6 as the likely cause of congenital stationary night blindness in a Tennessee Walking Horse.

BACKGROUND: The only known genetic cause of congenital stationary night blindness (CSNB) in horses is a 1378 bp insertion in TRPM1. However, an affected Tennessee Walking Horse was found to have no copies of this variant. OBJECTIVES: To identify the genetic cause for CSNB in an affected Tennessee Walking Horse. STUDY DESIGN: Case report detailing a whole-genome sequencing (WGS) approach to identify a causal variant. METHODS: A complete ophthalmic exam, including an electroretinogram (ERG), was performed on suspected CSNB-affected horse. WGS data were generated from the case and compared with data from seven other breeds (n = 29). One hundred candidate genes were evaluated for coding variants homozygous in the case and absent in all other horses. Protein modelling was used to assess the functional effects of the identified variant. A random cohort of 90 unrelated Tennessee Walking Horses and 273 horses from additional breeds were screened to estimate allele frequency of the GRM6 variant. RESULTS: ERG results were consistent with CSNB. WGS analysis identified a missense mutation in metabotropic glutamate receptor 6 (GRM6) (c.533C>T p.Thr178Met). This single nucleotide polymorphism (SNP) is predicted to be deleterious and protein modelling supports impaired binding of the neurotransmitter glutamate. This variant was not detected in 273 horses from three additional breeds. The estimated allele frequency in Tennessee Walking Horses is 10%. MAIN LIMITATIONS: Limited phenotype information for controls and no additional cases with which to replicate this finding. CONCLUSIONS: We identified a likely causal recessive missense variant in GRM6. Based on protein modelling, this variant alters GRM6 binding, and thus signalling from the retinal rod cell to the ON-bipolar cell, impairing vision in low light conditions. Given the 10% population allele frequency, it is likely that additional affected horses exist in this breed and further work is needed to identify and examine these animals.

The predictability of the Addiction Severity Index criminal justice assessment instrument and future imprisonment: A Swedish registry study with a national sample of adults with risky substance use.

OBJECTIVE: In Sweden, social workers uses the Addiction Severity Index (ASI) as their main assessment tool when assessing individuals with risky substance use (RSU) or substance use disorder. The aim of this study is to identify among individuals with RSU, the associations of ASI Composite Scores (CSs) with future imprisonment controlling for age, education level and gender. METHOD: Baseline ASI-data was merged with national registry data on prison sentences (2003-2016). Cox regression was used to estimate the associations between CSs for alcohol, drugs other than alcohol, legal, family and social relationships, employment, mental- and physical health and future imprisonment for adults (n = 14,914) assessed for RSU. RESULTS: The regression showed that all ASI CSs, age, education level and gender were significantly associated with imprisonment post ASI base-line assessment. The variables with the strongest association with imprisonment were ASI legal CS, followed by ASI drugs other than alcohol CS, ASI employment CS and being a male. ASI legal score showed the strongest association with imprisonment, with a 6 time increase in likelihood of imprisonment. DISCUSSION: Given the findings in this study, the strong significant association between ASI legal CS and future imprisonment, it seems as that the ASI-assessment instrument is a reliable and trustworthy assessment tool to use in clinical work. This should motivate social workers and other clinical health professionals to use and rely on the ASI assessment in their intervention planning for clients with RSU, to hopefully reduce future imprisonment and improve their social situation.

Associations between a risky psychosocial childhood and recurrent addiction compulsory care as adult.

BACKGROUND: Treatment for substance use disorder (SUD), results, in general, in improvements in terms of both drug use and social functioning. However, there are clients who are in need of repeated treatment. The aim of this retrospective study was to identify, for adults in compulsory care for severe SUD, the association between reporting having experienced a risky psychosocial childhood and repeated entries into the Swedish compulsory care system for SUD. METHOD: Hierarchical logistic regression and mediation analysis methods were used to analyse data from the Swedish National Board of Institutional Care (SiS) database. The sample included 2719 adults assessed at their compulsory care intake. The study examined the association between history of institutional care, family with SUD or psychiatric problem and repeated compulsory care entries as an adult controlling for main drug, age and gender. RESULTS: In the regression model the factor with the strongest association with repeated compulsory care intakes for SUD, was as a child having been in mandated institutional care (OR = 2.0 (1.60-2.51)). The proportion of the total effect that is mediated through LVU (law (1990:52) the care of young persons (special provisions) act) was 33% for SUD problems in family during childhood, 44% for psychiatric problems in family during childhood, and 38% for having been in foster care. CONCLUSION: Having been in mandated institutional care as a youth was strongly associated with repeated compulsory care for SUD as an adult. This is concerning since receipt of services as a child is supposed to mediate against the consequences of risky childhood conditions. These adults, as a group, are in need of a well-coordinated and integrated system of extensive aftercare services to reduce the likelihood of re-entry into compulsory care for an SUD.

A Third MLPH Variant Causing Coat Color Dilution in Dogs.

Altered melanosome transport in melanocytes, resulting from variants in the melanophilin (MLPH) gene, are associated with inherited forms of coat color dilution in many species. In dogs, the MLPH gene corresponds to the D locus and two variants, c.-22G > A (d1) and c.705G > C (d2), leading to the dilution of coat color, as described. Here, we describe the independent investigations of dogs whose coat color dilution could not be explained by known variants, and who report a third MLPH variant, (c.667_668insC) (d3), which leads to a frameshift and premature stop codon (p.His223Profs*41). The d3 allele is found at low frequency in multiple dog breeds, as well as in wolves, wolf-dog hybrids, and indigenous dogs. Canids in which the d3 allele contributed to the grey (dilute) phenotype were d1/d3 compound heterozygotes or d3 homozygotes, and all non-dilute related dogs had one or two D alleles, consistent with a recessive inheritance. Similar to other loci responsible for coat colors in dogs, this, alongside likely additional allelic heterogeneity at the D locus, or other loci, must be considered when performing and interpreting genetic testing.

Pigment Intensity in Dogs is Associated with a Copy Number Variant Upstream of KITLG.

Dogs exhibit a wide variety of coat color types, and many genes have been identified that control pigment production, appearance, and distribution. Some breeds, such as the Nova Scotia Duck Tolling Retriever (NSDTR), exhibit variation in pheomelanin pigment intensity that is not explained by known genetic variants. A genome-wide association study comparing light red to dark red in the NSDTR identified a significantly associated region on canine chromosome 15 (CFA 15:23 Mb-38 Mb). Coverage analysis of whole genome sequence data from eight dogs identified a 6 kb copy number variant (CNV) 152 kb upstream of KITLG. Genotyping with digital droplet PCR (ddPCR) confirmed a significant association between an increased copy number with the dark-red coat color in NSDTR (p = 6.1 × 10-7). The copy number of the CNV was also significantly associated with coat color variation in both eumelanin and pheomelanin-based Poodles (p = 1.5 × 10-8, 4.0 × 10-9) and across other breeds. Moreover, the copy number correlated with pigment intensity along the hair shaft in both pheomelanin and eumelanin coats. KITLG plays an important role in melanogenesis, and variants upstream of KITLG have been associated with coat color variation in mice as well as hair color in humans consistent with its role in the domestic dog.

The Associations between Risky Psychosocial Environment, Substance Addiction Severity and Imprisonment: A Swedish Registry Study.

Objective: Both childhood and adult psychosocial stressors have been identified as links to both increased risk for substance use disorder (SUD) and increased risk of imprisonment. The aim of this retrospective study is to identify, for a sample of 14,914 adults who all were assessed for risky substance use or a SUD, the importance of having a history of psychosocial stressors compared to current addiction severity. The analyses control for age, gender and education on the likelihood of future imprisonment. Method: Baseline Addiction Severity Index data (ASI) were merged with national registry data on prison sentences from 2003 to 2016. In the analysis, a Cox regression was used to study the association between independent variables and the likelihood of future imprisonment. Results: In the regression, five variables showed significant association to increased risk of imprisonment: ASI drugs other than alcohol Composite Score (positive relationship), ASI alcohol Composite Score (negative relationship), age (younger), education (lower) and parental problems with drugs other than alcohol. The factor with strongest association with imprisonment was the ASI drugs other than alcohol Composite Score, which showed the highest HR = 10.63 (3.50-32.31) for women and HR = 5.52 (3.77-8.08) for men to predict the likelihood of imprisonment. Discussion: Research is needed on why individuals with history of psychosocial stressors have a higher likelihood of imprisonment compared to their counterparts. Findings indicate that a high ASI Composite Score for drugs other than alcohol are strong predictors of future criminality and criminal justice system involvement.

Author Correction: Applications and efficiencies of the first cat 63 K DNA array.

A correction to this article has been published and is linked from the HTML and PDF versions of this paper. The error has been fixed in the paper.

Genetic heterogeneity and diversity of North American golden retrievers using a low density STR marker panel.

Thirty-three autosomal short tandem repeat (STR) markers were used to evaluate genetic heterogeneity and diversity in 525 golden retrievers (GRs). This breed was selected because of its popularity and artificial selection for conformation vs. performance phenotypes. Seven additional STRs were used to evaluate the highly polymorphic dog leukocyte antigen (DLA) class I and class II regions. From 3 to 13 alleles were found at each of the 33 loci (mean 7) and the average effective alleles (Ne) was 3.34. The observed heterozygosity was 0.65 and the expected heterozygosity was 0.68. The resulting fixation index was 0.035 indicating that the population was randomly breeding. We found that modern GRs retain 46% of genomic diversity present in all canids and 21/175 (12%) and 20/90 (22%) of the known DLA class I and class II haplotypes, respectively. Selection for performance or conformation led to a narrowing of genomic and DLA diversity with conformation having a greater effect than performance. A comparison was made between coefficient of inbreeding (COI) determined from 10 or 12 generation pedigrees and DNA based internal relatedness values. A weak but significant correlation was observed between IR score and 10 or 12 generation COI (r = 0.38, p<0.0001 and r = 0.40, p<0.0001, respectively). IR values were higher in conformation than performance lines but only significant at p = 0.17. This was supported by 10 and 12 generation COI values that were significantly (p<0.0001) higher in conformation than performance lines. We demonstrate herein that a low density of STR markers can be utilized to study the genetic makeup of GRs.

Author Correction: Applications and efficiencies of the first cat 63K DNA array.

A correction to this article has been published and is linked from the HTML and PDF versions of this paper. The error has been fixed in the paper.

Applications and efficiencies of the first cat 63K DNA array.

The development of high throughput SNP genotyping technologies has improved the genetic dissection of simple and complex traits in many species including cats. The properties of feline 62,897 SNPs Illumina Infinium iSelect DNA array are described using a dataset of over 2,000 feline samples, the most extensive to date, representing 41 cat breeds, a random bred population, and four wild felid species. Accuracy and efficiency of the array's genotypes and its utility in performing population-based analyses were evaluated. Average marker distance across the array was 37,741 Kb, and across the dataset, only 1% (625) of the markers exhibited poor genotyping and only 0.35% (221) showed Mendelian errors. Marker polymorphism varied across cat breeds and the average minor allele frequency (MAF) of all markers across domestic cats was 0.21. Population structure analysis confirmed a Western to Eastern structural continuum of cat breeds. Genome-wide linkage disequilibrium ranged from 50-1,500 Kb for domestic cats and 750 Kb for European wildcats (Felis silvestris silvestris). Array use in trait association mapping was investigated under different modes of inheritance, selection and population sizes. The efficient array design and cat genotype dataset continues to advance the understanding of cat breeds and will support monogenic health studies across feline breeds and populations.

Correction: A Novel Variant in CMAH Is Associated with Blood Type AB in Ragdoll Cats.

[This corrects the article DOI: 10.1371/journal.pone.0154973.].

Isolation and characterization of canine placenta-derived mesenchymal stromal cells for the treatment of neurological disorders in dogs.

Spinal cord injury (SCI) is a devastating disorder that affects humans and dogs. The prognosis of SCI depends on the severity of the injury and can include varying levels of motor and sensory deficits including devastating paraplegia and quadriplegia. Placental mesenchymal stromal cells (PMSCs) have been shown to improve wound healing and possess neuroprotective and immunomodulatory capabilities, but have not yet been clinically tested for the treatment of SCI. This study established a protocol to isolate fetal PMSCs from canine placentas and characterized their paracrine secretion profile and ability to stimulate neurons in vitro to assess their potential as a treatment option for neurological disorders in dogs. Canine PMSCs (cPMSCs) were plastic adherent and capable of trilineage differentiation. cPMSCs expressed typical MSC markers and did not express hematopoietic or endothelial cell markers. Genotyping of cPMSCs revealed fetal rather than maternal origin of the cells. cPMSCs were viable and mitotically expansive in a collagen hydrogel delivery vehicle, and they secreted the immunomodulatory and neurotrophic paracrine factors interleukin (IL)-6, IL-8, monocyte chemoattractant protein 1 (MCP-1), and vascular endothelial growth factor (VEGF). cPMSCs also stimulated the growth of complex neural networks when co-cultured with SH-SY5Y cells, a neuroblastoma cell line used to model neuron growth in vitro. cPMSCs are analogous to human PMSCs. They meet the criteria to be defined as MSCs and represent a potential regenerative therapy option for neurological disorders in dogs with their robust growth in collagen hydrogel, stimulation of neural network formation, and secretion of potent paracrine factors. © 2017 International Society for Advancement of Cytometry.