Independent COL17A1 Variants in Cats with Junctional Epidermolysis Bullosa.

Epidermolysis bullosa (EB), characterized by defective adhesion of the epidermis to the dermis, is a heterogeneous disease with many subtypes in human patients and domestic animals. We investigated two unrelated cats with recurring erosions and ulcers on ear pinnae, oral mucosa, and paw pads that were suggestive of EB. Histopathology confirmed the diagnosis of EB in both cats. Case 1 was severe and had to be euthanized at 5 months of age. Case 2 had a milder course and was alive at 11 years of age at the time of writing. Whole genome sequencing of both affected cats revealed independent homozygous variants in COL17A1 encoding the collagen type XVII alpha 1 chain. Loss of function variants in COL17A1 lead to junctional epidermolysis bullosa (JEB) in human patients. The identified splice site variant in case 1, c.3019+1del, was predicted to lead to a complete deficiency in collagen type XVII. Case 2 had a splice region variant, c.769+5G>A. Assessment of the functional impact of this variant on the transcript level demonstrated partial aberrant splicing with residual expression of wildtype transcript. Thus, the molecular analyses provided a plausible explanation of the difference in clinical severity between the two cases and allowed the refinement of the diagnosis in the affected cats to JEB. This study highlights the complexity of EB in animals and contributes to a better understanding of the genotype-phenotype correlation in COL17A1-related JEB

Canine NAPEPLD-associated models of human myelin disorders

Canine leukoencephalomyelopathy (LEMP) is a juvenile-onset neurodegenerative disorder of the CNS white matter currently described in Rottweiler and Leonberger dogs. Genome-wide association study (GWAS) allowed us to map LEMP in a Leonberger cohort to dog chromosome 18. Subsequent whole genome re-sequencing of a Leonberger case enabled the identification of a single private homozygous non-synonymous missense variant located in the highly conserved metallo-beta-lactamase domain of the N-acyl phosphatidylethanolamine phospholipase D (NAPEPLD) gene, encoding an enzyme of the endocannabinoid system. We then sequenced this gene in LEMP-affected Rottweilers and identified a different frameshift variant, which is predicted to replace the C-terminal metallo-beta-lactamase domain of the wild type protein. Haplotype analysis of SNP array genotypes revealed that the frameshift variant was present in diverse haplotypes in Rottweilers, and also in Great Danes, indicating an old origin of this second NAPEPLD variant. The identification of different NAPEPLD variants in dog breeds affected by leukoencephalopathies with heterogeneous pathological features, implicates the NAPEPLD enzyme as important in myelin homeostasis, and suggests a novel candidate gene for myelination disorders in people

Genomic Diversity and Runs of Homozygosity in Bernese Mountain Dogs

Bernese mountain dogs are a large dog breed formed in the early 1900s in Switzerland. While originally farm dogs that were used for pulling carts, guarding, and driving cattle, today they are considered multi-purpose companion and family dogs. The breed is predisposed to several complex diseases, such as histiocytic sarcoma, degenerative myelopathy, or hip dysplasia. Using whole-genome sequencing (WGS) data, we assessed the genomic architecture of 33 unrelated dogs from four countries: France, Sweden, Switzerland, and the United States. Analysis of runs of homozygosity (ROH) identified 12,643 ROH with an average length of 2.29 Mb and an average inbreeding coefficient of 0.395. Multidimensional scaling analysis of the genetic relatedness revealed limited clustering of European versus USA dogs, suggesting exchanges of breeding stock between continents. Furthermore, only two mtDNA haplotypes were detected in the 33 studied dogs, both of which are widespread throughout multiple dog breeds. WGS-based ROH analyses revealed several fixed or nearly fixed regions harboring discreet morphological trait-associated as well as disease-associated genetic variants. Several genes involved in the regulation of immune cells were found in the ROH shared by all dogs, which is notable in the context of the breed's strong predisposition to hematopoietic cancers. High levels of inbreeding and relatedness, strongly exaggerated in the last 30 years, have likely led to the high prevalence of specific genetic disorders in this breed

Условия получения и диэлектрические свойства СВЧ-керамики составов (1–x)((Mg0,2Zn0,8)TiO3–xCaTiO3

The results of investigation of the temperature and frequency dependences of the dielectric characteristics of microwave ceramics of (1–x)((Mg0,2Zn0,8)TiO3–xCaTiO3 ((1–x)(MZT)–xCT) (0.1 ≤ x< 0.6) compositions synthesized from a mixture of oxides (the first method) and from a mixture of a pre-prepared solid solution of (Mg0,2 Zn0,8)TiO3 and a CaTiO3  compounds (the second method), as well as ceramics of these compositions doped at the stage of sintering 1–2 % tin and tungsten are presented. It is shown that the synthesized ceramics is a composite consisting of a mixture of phases formed on the basis of solid solutions of (Zn, Mg)2 TiO4 , (Zn, Mg)TiO3  and the CaTiO3  compounds, the ratio of which in ceramics depends on the composition of the initial mixture and the synthesis conditions. It is established that the dielectric constant (ε) of ceramics synthe sized from a mixture of oxides increases with increasing CaTiO3 content in the (1–x)((Mg0,2Zn0,8)TiO3–xCaTiO3 system. This ceramics is characterized by small values of the temperature coefficient of dielectric constant (TKε ) and the dielectric loss tangent (tanδ) in the temperature range of 20–200 °C. For ceramics synthesized according to the second method, the high temperature stability of TKε and the small value of tanδ are observed in the temperature range of 20–150 °C. Doping ceramics with tin and tungsten oxides leads to an increase in ε and a decrease in dielectric losses.Представлены результаты исследований температурной и частотной зависимостей диэлектрических характеристик СВЧ-керамики составов (1–x)(Mg0,2 Zn0,8)TiO3 –xCaTiO3 ((1–x)(MZT)–xCT) (0,1 ≤x <0,6), синтезированной из смеси оксидов (первый способ) и из смеси предварительно полученных твердого раствора (Mg0,2 Zn0,8)TiO3 и соединения CaTiO3 (второй способ), а также керамик данных составов, допированных на стадии спекания 1–2 % олова и вольфрама. Показано, что синтезированная керамика представляет собой композит, состоящий из смеси фаз, образующихся на основе твердых растворов (Zn, Mg)2 TiO4 , (Zn, Mg)TiO3  и соединения CaTiO3 , соотношение которых в керамике зависит от состава исходной шихты и условий синтеза. Установлено, что диэлектрическая проницаемость (ε) керамики, синтезированной из смеси оксидов, увеличивается с увеличением содержания CaTiO3  в системе (1–x)(Mg0,2 Zn0,8)TiO3 –xCaTiO3. Данная керамика характеризуется малыми значениями температурного коэффициента диэлектрической проницаемости (ТКε ) и тангенса угла диэлектрических потерь (tgδ) в области температур 20–200 °C.Для керамики, синтезированной по второму способу, высокая температурная стабильность ТКε и малое значение tgδ наблюдаются в области температур 20–150 °C. Допирование керамик оксидами олова и вольфрама приводит к увеличению ε и снижению диэлектрических потерь

ВЛИЯНИЕ УЛЬТРАЗВУКОВОГО ВОЗДЕЙСТВИЯ НА КРИСТАЛЛИЧЕСКУЮ СТРУКТУРУ КЕРАМИКИ НА ОСНОВЕ ЦИРКОНАТА-ТИТАНАТА СВИНЦА

The crystal structure and electrical properties of ceramics obtained from sonicated powders of lead zirconate-titanate were investigated. It is found that ultrasonic treatment of the initial powders allows to obtain piezoelectric ceramics with the monocrystalline structure and desired physical properties.Исследованы кристаллическая структура и электрофизические свойства керамик, полученных из обработанных ультразвуком порошков цирконата-титаната свинца. Установлено, что ультразвуковое воздействие на исходные порошки дает возможность получать пьезоэлектрическую керамику с монокристаллической структурой и заданными физическими свойствами

Identification of ocular cicatricial pemphigoid antibody binding site(s

PURPOSE. To identify specific site(s) on human ␤4 molecule to which sera from ocular cicatricial pemphigoid (OCP) patients bind and to determine its role in the process of blister formation. METHODS. Clone the fragments representing the extracellular and intracellular domain of ␤4 molecule from normal human conjunctival mRNA into an expression vector; map the region to which sera from OCP patients bind by Western blot analysis. Determine the role of the immunodominant region in pathogenesis by demonstrating the ability of the rabbit antibody to the immunodominant region to produce separation of basement membrane zone (BMZ) from the basal epithelial layer when incubated with normal human conjunctiva in an in vitro organ culture model. RESULTS. Majority of the OCP sera tested bound to the Cterminal end of the intracellular domain (IC3.0) of the human ␤4 integrin. Further subcloning of IC3.0 demonstrated that a smaller fragment extending from 1489 aa to 1572 aa (IC3.4) was responsible for this binding. This region may have multiple antibody binding sites. Antibody to human IC3.0 and IC3.4 produced in rabbit, resulted in BMZ separation, histologically identical with that observed when normal human conjunctiva was cultured with OCP sera in an human conjunctival organ culture model. CONCLUSIONS. These observations identify IC3.4 as the antibody binding site for sera of OCP patients and suggest a possible role for it in blister formation. Indirectly it highlights certain important aspects of the structural and functional dynamics of the biology of the hemidesmosomes and basement membranes. (Invest Ophthalmol Vis Sci. 2001;42:379 -385) M ucous membrane pemphigoid (MMP) is a multisystemic autoimmune inflammatory disease that affects mucous membrane derived from stratified squamous epithelium including the conjunctiva and occasionally the skin. 1,2 In some patients, the involvement of the conjunctiva is more prominent than other mucous membranes. Such a subset of MMP patients are referred to as having ocular cicatricial pemphigoid (OCP). Progressive subepithelial fibrosis follows the chronic conjunctivitis, resulting in severe dryness of the eye, ocular keratinization, and blindness secondary to corneal scarring. Sera of patients with subepithelial blistering diseases have demonstrable levels of circulating antibodies that bind to different antigens within the basement membrane zone (BMZ) of skin and mucosa. 10 There are several human autoimmune diseases in which the target autoantigens are identified as being intracellular in location. MATERIALS AND METHODS Sera The method section confirms adherence to the Declaration of Helsinki. Sera used in this study were obtained from 20 patients with active mucous membrane pemphigoid before beginning of systemic treatment. These patients had the pemphigoid disease process involving multiple mucosa but not the skin. Ocular involvement was the most prominent symptom, resulting in blindness in many of these patients. The clinical diagnosis of OCP was established by routine histology and confirmed by direct immunofluorescence of the conjunctiva. The presence of IgG and or complement was detected in the conjunctival BMZ. Sera of these OCP patients binds to the epidermal side of the salt split skin. Control sera were obtained from 20 healthy individuals, 5 patients each with confirmed bullous pemphigoid (BP) and pemphigus vulgaris (PV). Blood samples were collected after informed consent, and the study was approved by the institutional review board

A Missense Variant Affecting the C-Terminal Tail of UNC93B1 in Dogs with Exfoliative Cutaneous Lupus Erythematosus (ECLE)

Cutaneous lupus erythematosus (CLE) in humans encompasses multiple subtypes that exhibit a wide array of skin lesions and, in some cases, are associated with the development of systemic lupus erythematosus (SLE). We investigated dogs with exfoliative cutaneous lupus erythematosus (ECLE), a dog-specific form of chronic CLE that is inherited as a monogenic autosomal recessive trait. A genome-wide association study (GWAS) with 14 cases and 29 controls confirmed a previously published result that the causative variant maps to chromosome 18. Autozygosity mapping refined the ECLE locus to a 493 kb critical interval. Filtering of whole genome sequence data from two cases against 654 controls revealed a single private protein-changing variant in this critical interval, UNC93B1:c.1438C>A or p.Pro480Thr. The homozygous mutant genotype was exclusively observed in 23 ECLE affected German Shorthaired Pointers and an ECLE affected Vizsla, but absent from 845 controls. UNC93B1 is a transmembrane protein located in the endoplasmic reticulum and endolysosomes, which is required for correct trafficking of several Toll-like receptors (TLRs). The p.Pro480Thr variant is predicted to affect the C-terminal tail of the UNC93B1 that has recently been shown to restrict TLR7 mediated autoimmunity via an interaction with syndecan binding protein (SDCBP). The functional knowledge on UNC93B1 strongly suggests that p.Pro480Thr is causing ECLE in dogs. These dogs therefore represent an interesting spontaneous model for human lupus erythematosus. Our results warrant further investigations of whether genetic variants affecting the C-terminus of UNC93B1 might be involved in specific subsets of CLE or SLE cases in humans and other species

A Missense Variant Affecting the C-Terminal Tail of UNC93B1 in Dogs with Exfoliative Cutaneous Lupus Erythematosus (ECLE)

Cutaneous lupus erythematosus (CLE) in humans encompasses multiple subtypes that exhibit a wide array of skin lesions and, in some cases, are associated with the development of systemic lupus erythematosus (SLE). We investigated dogs with exfoliative cutaneous lupus erythematosus (ECLE), a dog-specific form of chronic CLE that is inherited as a monogenic autosomal recessive trait. A genome-wide association study (GWAS) with 14 cases and 29 controls confirmed a previously published result that the causative variant maps to chromosome 18. Autozygosity mapping refined the ECLE locus to a 493 kb critical interval. Filtering of whole genome sequence data from two cases against 654 controls revealed a single private protein-changing variant in this critical interval, UNC93B1:c.1438C>A or p.Pro480Thr. The homozygous mutant genotype was exclusively observed in 23 ECLE affected German Shorthaired Pointers and an ECLE affected Vizsla, but absent from 845 controls. UNC93B1 is a transmembrane protein located in the endoplasmic reticulum and endolysosomes, which is required for correct trafficking of several Toll-like receptors (TLRs). The p.Pro480Thr variant is predicted to affect the C-terminal tail of the UNC93B1 that has recently been shown to restrict TLR7 mediated autoimmunity via an interaction with syndecan binding protein (SDCBP). The functional knowledge on UNC93B1 strongly suggests that p.Pro480Thr is causing ECLE in dogs. These dogs therefore represent an interesting spontaneous model for human lupus erythematosus. Our results warrant further investigations of whether genetic variants affecting the C-terminus of UNC93B1 might be involved in specific subsets of CLE or SLE cases in humans and other species

Two MC1R loss-of-function alleles in cream-coloured Australian Cattle Dogs and white Huskies.

Loss-of-function variants in the MC1R gene cause recessive red or yellow coat-colour phenotypes in many species. The canine MC1R:c.916C>T (p.Arg306Ter) variant is widespread and found in a homozygous state in many uniformly yellow- or red-coloured dogs. We investigated cream-coloured Australian Cattle Dogs whose coat colour could not be explained by this variant. A genome-wide association study with 10 cream and 123 red Australian Cattle Dogs confirmed that the cream locus indeed maps to MC1R. Whole-genome sequencing of cream dogs revealed a single nucleotide variant within the MITF binding site of the canine MC1R promoter. We propose to designate the mutant alleles at MC1R:c.916C>T as e1 and at the new promoter variant as e2. Both alleles segregate in the Australian Cattle Dog breed. When we considered both alleles in combination, we observed perfect association between the MC1R genotypes and the cream coat colour phenotype in a cohort of 10 cases and 324 control dogs. Analysis of the MC1R transcript levels in an e /e compound heterozygous dog confirmed that the transcript levels of the e2 allele were markedly reduced with respect to the e1 allele. We further report another MC1R loss-of-function allele in Alaskan and Siberian Huskies caused by a 2-bp deletion in the coding sequence, MC1R:c.816_817delCT. We propose to term this allele e3. Huskies that carry two copies of MC1R loss-of-function alleles have a white coat colour