Exclusion of NEU1 and PPGB from candidate genes for a lysosomal storage disease in Japanese Black cattle

A case of lysosomal storage disease has been reported in a calf of Japanese Black cattle. Lysosomal storage diseases are hereditary diseases caused by deficiency of lysosomal hydrolases. The clinical and pathological features and accumulated substrates of the affected animal indicated a possibility of sialidosis or galactosialidosis caused by deficiency of neuraminidase (NEU1) or protective protein for β-galactosidase (PPGB). In the present study, we investigated nucleotide sequences of the genes encoding these two proteins to evaluate whether mutation of these genes is involved in this disease. We determined cattle genomic sequences of these two genes by using bovine EST sequences and the nucleotide sequences of all exons of these genes were compared between affected and normal animals. The results showed several nucleotide substitutions, but none of them was a functional mutation or specific to the affected animal. Furthermore, genotyping of the microsatellite markers in the vicinity of these two genes revealed no homozygosity of the chromosomal regions including these genes in the affected animal. These findings indicated that neither NEU1 nor PPGB gene is responsible for the lysosomal storage disease of Japanese Black cattle and therefore the disease is neither sialidosis nor galactosialidosis.     

Lysosomal storage diseases in dogs and cats

Lysosomal storage diseases are inborn errors of metabolism which are common in a number of breeds of dog and cat, due to specific, genetically determined, deficiencies of degradative enzymes. They are generally inherited as autosomal recessive traits. The enzyme substrate may be a complex lipid, glycoprotein or polysaccharide. In enzyme deficiency this substrate accumulates in swollen lysosomes in specific organs. Assay of the enzyme can be employed to detect affected individuals and also heterozygotes. It is possible to control these diseases by the identification of heterozygous animals and refraining from using them for breeding. Affected animals are normal at birth but the onset of disease is generally during the first months of life. The diseases are progressive and fatal. Neurological signs, hepato- and splenomegaly and skeletal abnormalities are commonly seen.     

The gangliosidoses: comparative features and research applications.

Ganglioside storage diseases are inherited defects of lysosomal hydrolases that result in intralysosomal accumulation of gangliosides and other complex metabolites. Gangliosidoses occur in man, cats, cattle, dogs and swine. In all species, these diseases are characterized clinically by relentlessly progressive neurological deterioration. Lysosomal hypertrophy with characteristic ultrastructural inclusions occur in neurons, endothelial and other cells. Definitive diagnosis requires biochemical identification of the storage product and enzyme deficiency. Gangliosidoses in animals are important models of human lysosomal diseases and may be a significant complication in the maintenance of certain purebred stocks of domestic animals.     

Glycogenosis type III in the dog.

Enzyme and glycogen structure studies have been carried out on tissues of a glycogenotic dog, the clinical and pathological characteristics of which are reported in the accompanying paper. Liver glucose-6-phosphatase, leukocyte and liver acid maltase, and liver and skeletal muscle glycogen Phosphorylase all appeared largely unaffected. The activity of the muscle and liver debranching enzyme (amylo-l,6-glucosidase), determined by two independent assay methods, was, however, reduced to between 0 and 7 % of normal activity. Glycogen structure studies with Phosphorylase or iodine spectra revealed that the abnormally large amounts of glycogen found in liver and skeletal muscle had abnormally short branches, as would be expected for a deficiency of debranching enzyme. It is thus clear that the dog had suffered from the equivalent of Cori''s disease (limit dextrinosis, type III glycogen storage disease). Preliminary data indicate that it may be possible to identify heterozygotes based on a study of the debranching enzyme of leukocytes.     

Canine GM2‐Gangliosidosis Sandhoff Disease Associated with a 3‐Base Pair Deletion in the HEXB Gene

Background

GM2‐gangliosidosis is a fatal neurodegenerative lysosomal storage disease (LSD) caused by deficiency of either β‐hexosaminidase A (Hex‐A) and β‐hexosaminidase B (Hex‐B) together, or the GM2 activator protein. Clinical signs can be variable and are not pathognomonic for the specific, causal deficiency.

Objectives

To characterize the phenotype and genotype of GM2‐gangliosidosis disease in an affected dog.

Animals

One affected Shiba Inu and a clinically healthy dog.

Methods

Clinical and neurologic evaluation, brain magnetic resonance imaging (MRI), assays of lysosomal enzyme activities, and sequencing of all coding regions of HEXA, HEXB, and GM2A genes.

Results

A 14‐month‐old, female Shiba Inu presented with clinical signs resembling GM2‐gangliosidosis in humans and GM1‐gangliosidosis in the Shiba Inu. Magnetic resonance imaging (MRI) of the dog's brain indicated neurodegenerative disease, and evaluation of cerebrospinal fluid (CSF) identified storage granules in leukocytes. Lysosomal enzyme assays of plasma and leukocytes showed deficiencies of Hex‐A and Hex‐B activities in both tissues. Genetic analysis identified a homozygous, 3‐base pair deletion in the HEXB gene (c.618‐620delCCT).

Conclusions and Clinical Importance

Clinical, biochemical, and molecular features are characterized in a Shiba Inu with GM2‐gangliosidosis. The deletion of 3 adjacent base pairs in HEXB predicts the loss of a leucine residue at amino acid position 207 (p.Leu207del) supporting the hypothesis that GM2‐gangliosidosis seen in this dog is the Sandhoff type. Because GM1‐gangliosidosis also exists in this breed with almost identical clinical signs, genetic testing for both GM1‐ and GM2‐gangliosidosis should be considered to make a definitive diagnosis.     

Clinical and pathologic features of ceroid lipofuscinosis in two Australian cattle dogs

A lysosomal storage disease was diagnosed in 2 Australian Cattle Dog siblings, using light and electron microscopic evaluation. Both dogs developed clinical signs of disease at about 1 year of age. Vision and motor function deteriorated over several months; by 2 years of age, the dogs were blind and had progressive ataxia. Cytoplasmic inclusions with ultrastructural patterns characteristic of ceroid lipofuscin were observed in most neurons examined and in the cells of several other parenchymatous tissues. Biochemical studies, including determination of lysosomal enzyme activities, excluded several other lysosomal storage diseases. In these dogs, the clinical and pathologic features of the disease were similar to those of the juvenile subtype of ceroid lipofuscinosis (Batten disease) in human beings.     

Equine diseases caused by known genetic mutations

The recent development of equine genome maps by the equine genome community and the complete sequencing of the horse genome performed at the Broad Institute have accelerated the pace of genetic discovery. This review focuses on genetic diseases in the horse for which a mutation is currently known, including hyperkalemic periodic paralysis, severe combined immunodeficiency, overo lethal white syndrome, junctional epidermolysis bullosa, glycogen branching enzyme deficiency, malignant hyperthermia, hereditary equine regional dermal asthenia, and polysaccharide storage myopathy. Emphasis is placed on the prevalence, clinical signs, etiology, diagnosis, treatment and prognosis for each disease.     

Mucolipidosis type II in a domestic shorthair cat

A seven-month-old, female domestic shorthair cat was presented to the Veterinary Teaching Hospital, University of Zurich, with abnormal facial features, retarded growth and progressive hindlimb paresis. On physical examination the cat had a flat, broad face with hypertelorism, frontal bossing, small ears and thickened upper and lower eyelids. The corneas of both eyes were clear and the pupils were dilated. The skin was generally thickened, most prominently on the dorsal aspect of the neck. Radiography of the entire skeleton revealed a severely deformed spinal column, bilateral hip luxation with hip dysplasia, an abnormally shaped skull and generalised decreased bone opacity. The clinical features and radiographic changes were suggestive of mucopolysaccharidosis. The toluidine blue spot test on a urine sample, however, was negative for glycosaminoglycans. Further biochemical investigations revealed a deficiency of the enzyme N-acetylglucosamine-1-phosphotransfer-ase (GlcNAc-phosphotransferase, EC 2.7.8.17) in peripheral leukocytes and an elevation of many lysosomal enzymes in the serum of the cat which is diagnostic for mucolipidosis type II. Histology and electron microscopy of different tissues are briefly summarised. The findings of this cat, the first reported case of mucolipidosis type II are compared with other similar storage diseases described in the cat.     

Mucopolysaccharidosis type VII in a German Shepherd Dog

A 12-week-old male German Shepherd Dog was evaluated because of a 3-week history of a progressive inability to ambulate. Clinical and laboratory findings included skeletal deformities, corneal cloudiness, cytoplasmic granules in the neutrophils and lymphocytes of blood and CSF and glycosaminoglycans in a urine sample (detected via a toluidine blue spot test). Enzyme activity and DNA analyses confirmed mucopolysaccharidosis type VII as a result of severe beta-glucuronidase deficiency; this condition had been identified in a mixed-breed dog (likely of German Shepherd Dog descent) that was reported 20 years earlier and caused by the same missense mutation. Because of the progressive nature of this disease, the puppy was euthanatized and all tissues examined contained evidence of lysosomal storage leading to marked cellular distention. Mucopolysaccharidosis type VII is only one of many hereditary lysosomal storage diseases identified in companion animals. It is important that clinicians recognize the typical signs of lysosomal storage diseases and are aware of the cytologic and urinary screening tests for mucopolysaccharidosis disorders. Furthermore, there are specific blood enzyme and DNA-based tests to distinguish the forms of mucopolysaccharidosis in affected and carrier animals.     

Glycogen branching enzyme deficiency in quarter horse foals

Seven related Quarter Horse foals that died by 7 weeks of age were examined for glycogen branching enzyme (GBE) deficiency. Clinical signs varied from stillbirth, transient flexural limb deformities, seizures, and respiratory or cardiac failure to persistent recumbency. Leukopenia (5 of 5 foals) as well as high serum creatine kinase (CK; 5 of 5), aspartate transaminase (AST; 4 of 4), and gamma glutamyl transferase (GGT; 5 of 5) activities were present in most foals, and intermittent hypoglycemia was present in 2 foals. Gross postmortem lesions were minor, except for pulmonary edema in 2 foals. Muscle, heart, or liver samples from the foals contained abnormal periodic acid Schiff's (PAS)-positive globular or crystalline intracellular inclusions in amounts proportional to the foal's age at death. Accumulation of an unbranched polysaccharide in tissues was suggested by a shift in the iodine absorption spectra of polysaccharide isolated from the liver and muscle of affected foals. Skeletal muscle total polysaccharide concentrations were reduced by 30%, but liver and cardiac muscle glycogen concentrations were normal. Several glycolytic enzyme activities were normal, whereas GBE activity was virtually absent in cardiac and skeletal muscle, as well as in liver and peripheral blood cells of affected foals. GBE activities in peripheral blood cells of dams of affected foals and several of their half-siblings or full siblings were approximately 50% of controls. GBE protein in liver determined by Western blot was markedly reduced to absent in affected foals, and in a half-sibling of an affected foal, it was approximately one-half the amount of normal controls. Pedigree analysis also supported an autosomal recessive mode of inheritance. The affected foals have at least 2,600 half-siblings. Consequently, GBE deficiency may be a common cause of neonatal mortality in Quarter Horses that is obscured by the variety of clinical signs that resemble other equine neonatal diseases.     

Clinical and Clinicopathologic Characteristics of Ovine GM-1 Gangliosidosis

Ovine GM-1 gangliosidosis is an inherited lysosomal storage disease. Nine lambs affected with the disease were studied to characterize clinical signs and to determine if there were any pathognomonic clinicopathologic abnormalities. Evaluation included physical, ophthalmic, and neurologic examinations, complete blood counts, serum enzyme and electrolyte analyses, urinalyses, cerebrospinal fluid analyses, blood gas analyses, roentgenograms, electromyograms, and electrocardiograms. Two affected lambs had clinicopathologic tests performed before and after the onset of clinical signs. The only consistent abnormalities recognized were nonspecific signs referable to the central nervous system; predominantly ataxia, conscious proprioceptive deficit most severe in the hind limbs, blindness, and recumbency. Lambs continued to eat and drink, though at diminished levels and with loss of body condition. It was concluded that there are no pathognomonic clinicopathologic abnormalities associated with ovine GM-1 gangliosidosis, and antemortem diagnosis requires enzyme assay of leukocytes or cultured fibroblasts, or lectin histochemistry of tissues obtained by biopsy. Lysosomal storage diseases should be considered among the differential diagnoses in young animals presenting with early neonatal death or with nonspecific neurological signs, in concert with an absence of diagnostic clinicopathologic findings.     

Congenital diseases of feline muscle and neuromuscular junction

Although muscle diseases occur relatively rarely in cats, a number of congenital feline myopathies have been described over the last 20 years and are reviewed in this paper. Some of them have been reported exclusively in specific breeds, including the hypokalaemic myopathy of Burmese cats, type IV glycogen storage disease in Norwegian Forest cats, or the myopathy of Devon Rex. Other congenital disorders of muscle and neuromuscular junction such as myotonia congenita, dystrophin-deficient hypertrophic feline muscular dystrophy, laminin alpha2 deficiency, or congenital myasthenia gravis may occur in any cat. A systematic approach is essential in order to efficiently obtain a timely diagnosis in cats showing signs of muscle disease. After a thorough clinical examination, this approach includes blood analyses (eg, serum concentration of muscle enzymes), electrophysiology where available (electromyography, nerve conduction studies), and sampling of muscle biopsies for histological, histochemical and immunohistochemical evaluation. When available, detection of healthy carriers of these genetic disorders is important to eliminate the gene mutations from breeding families. Clinicians regularly receiving feline patients must have a good knowledge of congenital feline myopathies and the features which enable a diagnosis to be made and prognosis given. Besides preserving or restoring the well-being of the myopathic patient, rapid and efficient information and counselling of the breeders are of central importance in order to prevent the recurrence of the problem in specific breeding lines.     

A variant form of neuronal ceroid lipofuscinosis in American bulldogs

Neuronal ceroid lipofuscinoses (NCLs) are inherited neurodegenerative diseases characterized by accumulations of autofluorescent lipopigments within cells of the nervous system. Nine related American Bulldogs demonstrated dysmetria in all limbs and paraparesis. Nonambulatory tetraparesis was observed only in the later stages of the disease. The clinical signs developed between 1 and 3 years of age and were slowly progressive over several years, which is inconsistent with most reports in other breeds. Results from blood tests for 8 different lysosomal storage diseases on 4 affected and 6 related but unaffected dogs were negative. Four affected dogs were euthanized and histopathologic examinations showed diffuse accumulations of periodic acid-Schiff-positive inclusions in neurons and axonal spheroids along the entire neuraxis and retinae. The most severe lesions were in the brainstem proprioceptive nuclei and spinal cord, consistent with clinical signs. The storage material was autofluorescent and immunohistochemically positive for products of lipid peroxidation. Ultrastructural analysis was consistent with NCL. Pedigree analysis supports an autosomal-recessive mode of inheritance. NCL has not been previously reported in the American Bulldog and these findings suggest a variant form of the canine disease.     

Anemia, splenomegaly, and increased osmotic fragility of erythrocytes in Abyssinian and Somali cats

OBJECTIVE: To determine clinical and clinicopathologic features of a chronic intermittent severe hemolytic anemia characterized by erythrocyte osmotic fragility in Abyssinian and Somali cats. DESIGN: Case series. ANIMALS: 13 Abyssinian and 5 Somali cats. PROCEDURES: History, pedigree information, and results of routine laboratory tests, special erythrocyte studies, and histologic evaluation of splenic and hepatic specimens were analyzed. RESULTS: Age at which clinical signs of anemia were first apparent ranged from 6 months to 5 years. Ten cats had splenomegaly. Most often, the PCV was between 15 and 25%, but it was as low as 5% at some times. The anemia was characterized by macrocytosis and mild to moderate reticulocytosis, but no poikilocytosis. Hyperglobulinemia, lymphocytosis, mild hyperbilirubinemia, and high hepatic enzyme activities were common findings. Results of Coombs tests and tests for infectious diseases were negative. The erythrocytic osmotic fragility was high in affected cats (mean osmotic fragility, 0.66 to 0.78%), compared with healthy cats (0.48 to 0.58). No specific membrane protein abnormality, erythrocyte enzyme deficiency, or hemoglobinopathy was identified. Histologic evaluation of splenic and hepatic specimens revealed extramedullary hematopoiesis and hemosiderosis. Four of the 5 Somali cats were closely related. CONCLUSIONS AND CLINICAL RELEVANCE: On the basis of results of pedigree analyses, the apparent breed predilection, and the exclusion of other known causes of anemia in cats, we believe that the hemolytic anemia in these cats was likely a result of a novel hereditary erythrocyte defect. A genetic predisposition to immune-mediated destruction of erythrocytes could not be ruled out.     

自由基与疾病研究进展

随着基础医学和生命科学的不断发展,人们对自由基的研究越来越多,其中就有大量关于自由基与疾病的研究。自由基作为机体的正常代谢产物,在平衡状态下,其在抗菌、消炎和抑制肿瘤等方面具有重要作用和意义;一旦平衡被打破,如机体受到疾病或某些外源性药物和毒物的侵害,自由基便会产生强大的伤害作用,造成生物膜的脂质过氧化损伤,引起酶、氨基酸、蛋白质的氧化破坏,对内脏器官、免疫系统的形态功能产生影响,从而引起机体疾病,甚至死亡。目前,研究发现很多疾病的发生发展都与自由基有关。文章就自由基的产生、种类、与疾病的关系及清除进行了综述。     

Acquired multiple acyl-CoA dehydrogenase deficiency and marked selenium deficiency causing severe rhabdomyolysis in a horse

This report describes a case of severe rhabdomyolysis in a pregnant mare associated with histopathologic and biochemical features of both selenium deficiency and acquired multiple acyl-CoA dehydrogenase deficiency (MADD) due to seasonal pasture myopathy (SPM). This case highlights the importance of assessing plasma selenium levels in horses with clinical signs of pasture myopathy as this deficiency may be a contributing or exacerbating factor.     

Investigations on manifestations of vitamin E and selenium deficiency in sheep and goats]

In a prospective study from 1991-1998 plasma concentrations of alpha-Tocopherole (VitE) and Selenium (Se) were analysed in 125 sheep and 32 goats with generalised motor disturbances or elevated plasma-activities of Creatine-Kinase (CK). VitE-values < 1.0 mg/l and Se-values < 0.08 mg/l were regarded as deficiency. Diagnosis of pathological manifestations was based on blood enzyme values. CK-values in plasma > 300 U/l or Aspatate-Amino-Transferase (ASAT) > 150 U/l were regarded as myopathy, Glutamat-Dehydrogenase-values > 25 U/l indicated a hepatopathy. 92 (74%) sheep and 10 (31%) goats showed VitE and/or Se deficiency. There were no principal differences in the frequencies of myopathies (67%) and hepatopathies (46%) between pure VitE deficiencies or pure Se deficiencies or combinations of both deficiencies. In VitE deficiency and in the combinations of VitE and Se deficiency the elevations of the enzyme activities were more pronounced than in Se deficiency. The lethality was significantly higher in combined VitE + Se deficiencies (73%) than in pure VitE or Se deficiencies (35%). Combined VitE + Se deficiencies occurred more frequent in lambs (53%) and was often accompanied by anaemia (29%) and hypoproteinaemia (32%). The diagnostic reliability of ASAT and GLDH for the detection of VitE and Se deficiency was better than CK. Diagnostic and therapeutic strategies in flock diseases are discussed.     

Glycogen storage disease type II in the Lapland dog

A newly recognized inherited metabolic disease in the Lapland dog is described. The metabolic defect is a deficiency of acid-alpha-glucosidase, a lysosomal hydrolase. The clinical picture is dominated by vomiting related to megaoesophagus, and progressive muscle weakness leading to exhaustion and death before two years of age. Cardiac abnormalities are observed. The main histopathologic lesion consists of glycogen accumulation, notably in membrane-bound vacuoles (glycogenosomes), involving all kinds of muscular tissue in particular. Recessive inheritance of the disease was demonstrated by complementation analysis. The enzyme protein is present in affected tissues, although in an inactive form. Based on the gene dosage phenomenon, an attempt was made to identify carrier dogs by means of a biochemical assay. Glycogen storage disease type II in the Lapland dog appears to be a homologous model for the infantile manifestation of glycogen storage disease type II (Pompe's disease) in man.     

Disorders of the vestibular system

Disorders of the vestibular system are common in veterinary practice and result in a clinical syndrome characterized by head tilt, asymmetric ataxia, and nystagmus. These signs may occur with lesions involving the peripheral or central vestibular structures. Careful neurologic evaluation of the animal and knowledge of surrounding neuroanatomic structures and their functions enable the clinician to differentiate between peripheral and central disorders. Conditions resulting in peripheral vestibular symptoms include idiopathic syndromes, otitis interna, trauma, neoplasia, drug-induced ototoxicity, and congenital disorders. Radiographs of the petrous temporal bone and tympanic bulla and examination of the tympanic membrane are useful in determining the specific etiology. Central vestibular dysfunction may result from inflammatory CNS diseases, neoplasia, trauma, thiamine deficiency, and storage diseases. Cerebrospinal fluid analysis, skull radiographs, and computed tomography are diagnostic tests useful in animals with central vestibular symptoms. Once the etiology is identified, specific therapy may be provided and a more accurate prognosis can be determined.     

Subgroups of canine antinuclear antibodies in relation to laboratory and clinical findings in immune-mediated disease

Background:  Autoimmune system diseases in dogs are commonly referred to as systemic lupus erythematosus (SLE), with a positive antinuclear antibody (ANA) test as a hallmark. In human patients, other systemic ANA-positive diseases with overlapping diagnostic features, referred to as SLE-related diseases, are described.  Objectives:  The objective of this study was to investigate whether different patterns of ANA reactivity represent different systemic autoimmune diseases in dogs. Methods:  Dogs with serum positive for ANA by indirect immunofluorescence (IIF-ANA, titer ≥1:100) (n=56) were identified retrospectively from the patient population at the Department of Small Animal Clinical Sciences, Swedish University of Agricultural Sciences. Dogs were grouped on the basis of ANA staining patterns, and the results of immunodiffusion tests. Clinical, hematologic, serum biochemical, radiologic, and pathologic examinations were described for each group.  Results:  Dogs with a chromosomal–positive, homogeneous ANA staining pattern (n=14) had clinical signs involving multiple organ systems; 8 dogs were anemic. Dogs with a speckled IIF-ANA staining pattern (n=42) primarily had clinical signs of musculoskeletal disorders, fatigue and fever. Precipitating antibodies by immunodiffusion were found only in dogs with a speckled IIF-ANA staining pattern and comprised 4 different subgroups based on antigen specificity. Conclusions: In dogs with homogeneous IIF-ANA staining, SLE is a probable diagnosis because of the diversity of clinical manifestations and autoantibody reactivity against chromosomal antigens. Dogs with a speckled IIF-ANA pattern may have SLE-related diseases, which, in turn, may be correlated with different immunodiffusion subgroups. These syndromes had overlapping clinicopathologic features, as described for human patients.