1,720,991 research outputs found
haploinsufficiency in a Holstein cattle with epilepsy
Full text linkIn this study, epilepsy with focal seizures progressing to generalized seizures was diagnosed in a 6-month-old Holstein heifer. The seizures were characterized by a brief pre-ictal phase with depression and vocalization. During the ictal phase eyelid spasms, tongue contractions, nodding and abundant salivation were observed, rapidly followed by a convulsive phase with bilateral tonic, clonic or tonic-clonic activity and loss of consciousness. Finally, during the postictal phase the heifer was obtunded and disorientated, unable to perceive obstacles and hypermetric, and pressed its head against objects. In the inter-seizure phase, the heifer was clinically normal. Neuropathology revealed axonal degeneration in the brainstem and diffuse astrocytic hypertrophic gliosis. Whole genome sequencing of the affected heifer identified a private heterozygous splice-site variant in DYRK1B (NM_001081515.1: c.-101-1G>A), most likely resulting in haploinsufficiency owing to loss-of-function. This represents a report of a DYRK1B-associated disease in cattle and adds DYRK1B to the candidate genes for epilepsy
First Report on Medical Treatment and Outcome of Burnt Cattle
Full text linkThe management of livestock affected by fire often comes down to two options: euthanasia or slaughtering. However, the therapeutic approach can be attempted for high-value cattle. The aim of a primary assessment is to identify signs of smoke inhalation injuries, cardiovascular impairment and shock and to determine the severity and extent of burn injuries. Full-thickness burns covering 40% or more of the body are highly unfavorable prognostic factors and are usually fatal. Moreover, it can take several days for the burns to appear in their full extent, leaving the prognosis uncertain. In this case report, the clinical findings, treatment and outcome of two burnt Holstein heifers are described. Daily wound care required cleaning, the removal of eschars and the application of topical antibacterial agents for seven months in order to discharge one heifer. The topical use of honey with a solution of povidone–iodine proved to be affordable and successful, with no residue risks. The other heifer was more severely wounded, and despite the administration of fluid therapy, pain management, anti-oxidants and anti-microbials, after initial stabilization, the animal’s condition worsened, leading to euthanasia. This confirms that the treatment of burnt cattle is possible but challenging due to the late onset of multi-organ failure
A frameshift mutation in MOCOS is associated with familial renal syndrome (xanthinuria) in Tyrolean Grey cattle
Full text linkBackground
Renal syndromes are occasionally reported in domestic animals. Two identical twin Tyrolean Grey calves exhibited weight loss, skeletal abnormalities and delayed development associated with kidney abnormalities and formation of uroliths. These signs resembled inherited renal tubular dysplasia found in Japanese Black cattle which is associated with mutations in the claudin 16 gene. Despite demonstrating striking phenotypic similarities, no obvious presence of pathogenic variants of this candidate gene were found. Therefore further analysis was required to decipher the genetic etiology of the condition.
Results
The family history of the cases suggested the possibility of an autosomal recessive inheritance. Homozygosity mapping combined with sequencing of the whole genome of one case detected two associated non-synonymous private coding variants: A homozygous missense variant in the uncharacterized KIAA2026 gene (g.39038055C > G; c.926C > G), located in a 15 Mb sized region of homozygosity on BTA 8; and a homozygous 1 bp deletion in the molybdenum cofactor sulfurase (MOCOS) gene (g.21222030delC; c.1881delG and c.1782delG), located in an 11 Mb region of homozygosity on BTA 24. Pathogenic variants in MOCOS have previously been associated with inherited metabolic syndromes and xanthinuria in different species including Japanese Black cattle. Genotyping of two additional clinically suspicious cases confirmed the association with the MOCOS variant, as both animals had a homozygous mutant genotype and did not show the variant KIAA2026 allele. The identified genomic deletion is predicted to be highly disruptive, creating a frameshift and premature termination of translation, resulting in severely truncated MOCOS proteins that lack two functionally essential domains. The variant MOCOS allele was absent from cattle of other breeds and approximately 4% carriers were detected among more than 1200 genotyped Tyrolean Grey cattle. Biochemical urolith analysis of one case revealed the presence of approximately 95% xanthine.
Conclusions
The identified MOCOS loss of function variant is highly likely to cause the renal syndrome in the affected animals. The results suggest that the phenotypic features of the renal syndrome were related to an early onset form of xanthinuria, which is highly likely to lead to the progressive defects. The identification of the candidate causative mutation thus enables selection against this pathogenic variant in Tyrolean Grey cattle
Human Brody disease and its animal model cattle pseudomyotonia: from understanding the pathogenetic mechanism to the identification of novel therapeutic approaches
No full textHuman Brody myopathy is a rare skeletal muscle disorder due to SERCA1 deficiency, resulting from mutations of ATP2A1 gene. The main clinical signs are exercise-induced stiffness and delayed muscular relaxation after physical exercises even mild, and muscles involved are prevalently voluntary muscles used for movement as legs, arms but also eyelids.
SERCA1 isoform is responsible for transporting Ca2+ from cytosol to the lumen of sarcoplasmic reticulum (SR) and controlling intracellular Ca2+ concentration. SERCA1 deficiency is responsible for impairment of muscle relaxation, after contraction, due to prolonged increase of calcium concentration in skeletal muscle fibers cytoplasm.
Our research group has demonstrated that Chianina cattle congenital pseudomyotonia represents the true counterpart of Brody myopathy. This is not surprising since, in the last years, the counterparts of human pathologies have been found in many domestic mammalian species. Interestingly, a missense mutation in the ATP2A1 gene is responsible for the pseudomyotonia phenotype and it has been clarified that the pathogenic mechanism underling pseudomyotonia involves ubiquitin-proteasome system degradation of the mutated bovine SERCA1.
At present, neither specific therapy nor mouse model for Brody myopathy exists. However, we have recently designed and proven in vitro a novel pharmacological approach based on the employment of protein folding correctors named CFTR (Cystic Fibrosis Transmembrane Regulator) exploited in Cystic Fibrosis, which help SERCA1 mutants to properly fold.
These data have been confirmed by those obtained ex vivo on adult skeletal muscle fibres from a biopsy from a pseudomyotonia affected subject. Moreover, local treatments of bovine pseudomyotonia muscle with the most effective in vitro CFTR corrector, although not easy, but show quite promising results
Brody Disease: A Novel Potential Therapeutic Approach for This Rare Human Disease
No full textBrody disease is a “rare” genetic disorder due to defects in SERCA1 gene and it is characterized by exercise-induced muscle stiffness and impairment of relaxation (Brody 1996). Bovine “congenital pseudomyotonia” (PMT) is a genetic muscular disorder very similar to human Brody myopathy for clinical signs. A missense mutation in the ATP2A1 gene (Drögemüller et al 2008), encoding sarco(endo)plasmic reticulum Ca2+-ATPase (SERCA1), causes congenital PMT in cattle and Brody disease in humans.
The mouse is the preferred animal model for preclinical evaluation of new therapeutic molecules. Unfortunately, it is not available for Brody disease (due to the prevalence of type II fibers in diaphragm muscle). Clinical symptoms genetic and biochemical findings clearly demonstrated that congenital PMT in Chianina cattle is the real analogue of Brody myopathy (Sacchetto et al 2009).
Our findings show that a SERCA1 mutation in bovine PMT results in a protein that is most likely misfolded, ubiquitinated, and destroyed prematurely by the ubiquitin-proteasome system, albeit retaining its catalytic capabilities (Bianchini et al. 2014).
The treatment with proteasome inhibitors restores SERCA levels and Ca2+ homeostasis in a cellular model and in muscle fibres from PMT affected animals (ex vivo experiments) (collected in conformance with the institutional guidelines for the care and use of animals).
At present, neither specific therapy nor mouse model for Brody myopathy exists.
However, a novel pharmacological approach based on the use of protein folding correctors known as CFTR (Cystic Fibrosis Transmembrane Regulator) explored in Cystic Fibrosis has recently been created and demonstrated in vitro. In a HEK293 cell model, treatment with CFTR correctors restored the expression level of mutated SERCA1.
These data have been confirmed in vivo by local treatments of bovine PMT muscle with the most effective in vitro CFTR corrector. (Pharmacological treatments and biopsies were authorized by the Italian Ministry of Health-authorization ID 1251/2021.) Besides, treatments with the same CFTR corrector on SERCA1 mutated zebrafish line Accordion, are ongoing
Human Brody disease and novel therapeutic approaches of its animal model cattle pseudomyotonia
No full textA Missense Variant in PLP2 in Holstein Cattle with X-Linked Congenital Mast Cell Tumor
Full text linkCongenital tumors occur infrequently in cattle. The aim of this study was to detail the clinicopathological phenotype of a Holstein calf with a congenital mast cell tumor and to identify the genetic cause by a whole-genome sequencing (WGS) trio-approach. An 18-day-old male Holstein calf was clinically examed and revealed multifocal, alopecic, thick and wrinkled skin lesions over the entire body. At 6 months of age, the general condition of the calf was characterized by retarded growth, poor nutritional status, and ulceration of the skin lesions. Histopathological examination revealed a primary cutaneous, poorly differentiated embryonal mast cell tumor with metastases in the lymph nodes and liver. Genetic analysis revealed a private X-linked variant in the PLP2 gene (chrX:87216480C > T; c.50C > T), which was present only in the genomes of the case (hemizygous) and his mother (heterozygous). It was absent in the sire as well as in 5365 control genomes. The identified missense variant exchanges the encoded amino acid of PLP2 at position 17 (p.Thr17Ile), which is classified as deleterious and affects a protein that plays a role in tumor growth and metastasis. Therefore, we suggested that the detected PLPL2 variant could be a plausible cause for this congenital condition in the affected calf
CATTLE CONGENITAL PSEUDOMYOTONIA AS ANIMAL MODEL: WHEN DIAPHRAGM MUSCLE CAN MAKE THE DIFFERENCE
No full textCattle “congenital pseudomyotonia” (PMT) is a muscular disorder described for the first time in
Chianina cattle breed [1] and subsequently in Romagnola breed, characterized by an impairment of
muscle relaxation induced by exercise. By DNA sequencing, we provided evidence of missense
mutations in ATP2A1 gene, coding for SERCA1 isoform. Pathological muscles of PMT affected
animals are characterized by a striking, selective reduction in the expression level of SERCA1
protein. Although present at low levels, the SERCA1 variants maintained the basic intrinsic properties
of wild-type (WT) SERCA1, notably the Ca2+ dependent ATPase activity [2,3]. On the bases of
clinical, genetic and biochemical analyses, our group has demonstrated that PMT is the true
counterpart of human Brody myopathy, a rare and orphan disease for which an appropriate
pharmacological treatment is now not available.
Our results showing that in bovine PMT, SERCA1 mutations do not abolish SERCA1 activity,
prompted us to hypothesize that the retention of functional properties of mutated SERCA pump
represents the requisite for the efficacy of a potential innovative therapy. We have recently designed
a novel pharmacological approach based on the employment of protein folding CFTR correctors,
specifically developed for rescuing type II Cystic Fibrosis Transmembrane Regulator (CFTR)
mutants [4].
Our preliminary results show that CFTR correctors successfully rescued in vitro SERCA1 mutants,
causing bovine PMT.
In vitro studies are fundamental for experimental settings and permit tight control of fundamental
principles and the testing of therapeutic hypotheses. They represent the first step toward a cure, being
the second step of preclinical research in drug development, represented by in vivo animal studies
which are essential and necessary before clinical trial can be pursued. Mouse is undoubtedly the
preferred model for preclinical therapeutic trials, nevertheless sometimes this model fails to mirror
the disease phenotype and therefore is unsuitable to study certain human pathologies. This is the case
of human Brody disease. Due to the wide difference in fiber type composition of diaphragm muscle
between mice and large mammals, including humans, mouse is not appropriate as animal model for
this pathology. To solve this problem, we treated locally bovine PMT affected muscle with CFTR
correctors, resulted from the most promising in vitro studies
Congenital Suborbital Undifferentiated Sarcoma in a Crossbred Calf
Full text linkUndifferentiated sarcomas are rare conditions that represent a group of unclassified sarcomas. The purpose of this study is to describe the clinical and pathological features of a calf showing a congenital infiltrating suborbital mass suggestive of undifferentiated sarcoma. The animal was referred because of respiratory distress and the presence of a right suborbital mass since birth. At ultrasonography, the mass displayed an irregular shape with multiple cavities. Radiographs revealed a diffuse, poorly defined mass with different densities overlying the bony structures of the skull. Endoscopy showed a co-involution of the mass in the right side with extension into the nasopharynx. Post-mortem examination showed a round, poorly demarcated neoplasia infiltrating the nasal turbinate and displacing the nasal septum. Histologically, the subcutis was expanded by lobules and bundles of densely cellular neoplastic spindle cells. The neoplasm infiltrated the underlying muscles, bone and the right retromandibular lymph node. The neoplastic cells had a diffuse intense cytoplasmic immunexpression to vimentin, and were negative to cytokeratin AE1/AE3, desmin, MUM1, IBA1, melan A, chromogranin and synaptophysin
Navel Healing and Calf Fitness for Transport
Full text linkDairy male calves are at risk of welfare compromise as they are usually transported at a very young age. The European Union has set a “completely healed navel” requirement for calf transport; moreover, a minimum age is established for longer journeys. However, this requirement has proven to be prone to misinterpretation. This study aimed to clarify what is meant by “navel healing” and to provide strong elements for reaching a consensus. The navels of 299 dairy calves (55 males, 244 females) aged 0–90 days were examined and scored 1 to 5 according to their healing status. Based on our results, a completely dry and shriveled navel (score 3) would imply a 25.5–38.0% risk of transporting too young calves. Alternatively, the presence of a scab covering the umbilical wound (score 4) would entail a 4.3% risk of transporting calves less than 10 days old and could be considered good practice for transporting calves (except for journeys exceeding 8 h). Conversely, complete navel healing (score 5) guarantees that calves that are too young are not transported; therefore, it should be considered best practice for transporting calves in general and the minimum requirement for transporting calves for journeys exceeding 8 h
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