RESUMO
Potentials of immortal proliferation and unlimited differentiation into all the three germ layers and germ cells in induced pluripotent stem cells (iPSCs) render them important bioresources for in vitro reconstitution and modeling of intravital tissues and organs in various animal models, thus contributing to the elucidation of pathomechanisms, drug discovery and stem cell-based regenerative medicine. We previously reported promising approaches for deriving transgene-free iPSCs from somatic fibroblasts of multiple mammalian species by episomal vector or RNA transfection, although the respective step-by-step protocols and the combinatorial usage of these methods, which achieved high induction efficiency, have not been described in the literature so far. Here, we provide a detailed step-by-step description of these methods with critical tips and slight modifications (improvements) to previously reported methods. We also report a novel method for the establishment of iPSCs from the Syrian hamster (also known as golden hamster; Mesocricetus auratus), a unique animal model of hibernation. We anticipate this methodology will contribute to stem cell biology and regenerative medicine research.
Assuntos
Células-Tronco Pluripotentes Induzidas , Animais , Diferenciação Celular , Reprogramação Celular , Fibroblastos , Mamíferos , TransgenesRESUMO
OBJECTIVE: To characterize the computed tomography (CT) findings for canine adrenal tumors, including cortical adenoma, cortical adenocarcinoma, and pheochromocytoma, and to evaluate the feasibility and usefulness of preoperative triple-phase helical CT for differentiation of tumor types and surgical planning. STUDY DESIGN: Retrospective study. ANIMALS: Dogs with adrenal tumors (n=36). METHODS: All dogs underwent triple-phase helical CT, followed by adrenalectomy and histopathological diagnosis of the resected mass. Precontrast images, arterial, venous, and delayed phase images were obtained. In all cases, morphological characteristics and CT values and calculations, including the percentage enhancement washout ratio, relative percentage washout, enhancement washin, and enhancement washout, were analyzed and compared among the tumor types. RESULTS: Of the 36 dogs with adrenal masses, cortical adenocarcinoma was most commonly diagnosed (16 dogs), followed by pheochromocytoma (13 dogs), and cortical adenoma (7 dogs). The precontrast minimum CT value and enhancement washout between venous and delayed phases in the cortical adenoma were significantly higher than those in the cortical adenocarcinoma. The maximum CT values of the precontrast image and arterial and venous phases, the enhancement washin and washouts, percentage enhancement washout ratio, and relative percentage washout in the pheochromocytomas were significantly higher than those in cortical adenocarcinoma. CONCLUSION: The differential diagnosis of canine adrenal tumors was feasible based on triple-phase CT findings, including morphological features, CT values, and intratumoral contrast attenuation. Preoperative diagnosis using triple-phase helical CT may be useful for surgical planning in dogs with adrenal tumors.
Assuntos
Neoplasias das Glândulas Suprarrenais/veterinária , Doenças do Cão/diagnóstico , Adenoma/diagnóstico , Adenoma/veterinária , Neoplasias das Glândulas Suprarrenais/diagnóstico , Adrenalectomia/veterinária , Carcinoma Adrenocortical/diagnóstico , Carcinoma Adrenocortical/veterinária , Animais , Meios de Contraste , Diagnóstico Diferencial , Doenças do Cão/diagnóstico por imagem , Doenças do Cão/cirurgia , Cães , Feminino , Masculino , Feocromocitoma/diagnóstico , Feocromocitoma/veterinária , Valor Preditivo dos Testes , Cuidados Pré-Operatórios/veterinária , Intensificação de Imagem Radiográfica , Estudos Retrospectivos , Tomografia Computadorizada Espiral/veterináriaRESUMO
The prevalences of displacement and deformity of the medial and lateral fabellae in dogs were investigated. This was a retrospective epidemiologic study. Radiographs of canine stifle joints (1022 limbs, 534 dogs) were obtained. The images were taken at the Nihon University Animal Medical Center and three private animal hospitals from January 2003 to July 2012. The position and morphology of the medial or lateral fabella were evaluated on the radiographs. The prevalence of displacement of the medial and lateral fabellae was 1.7% and 0.3%, respectively. The prevalence of deformity of the medial and lateral fabellae was 6.9% and 4.6%, respectively. Aplasia or hypoplasia of the fabella was detected more frequently in the medial fabella. On the other hand, a bipartite or multipartite fabella was observed more frequently in the lateral fabella. Nearly all cases of displacement or deformity of the fabella occurred in dogs weighing less than 10 kg. Abnormalities of the fabella were observed in the medial and lateral fabella. We found that abnormal fabellae were closely associated with medial patellar luxation and to a lesser extent with cranial cruciate ligament rupture. No clinical signs were associated with an abnormality of the fabella, with the exception of two dogs with traumatic avulsion of the lateral head of the gastrocnemius muscle.
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Doenças Ósseas/veterinária , Doenças do Cão/epidemiologia , Membro Posterior/patologia , Animais , Doenças Ósseas/congênito , Doenças Ósseas/epidemiologia , Doenças Ósseas/patologia , Doenças do Cão/patologia , Cães , Feminino , Japão/epidemiologia , Masculino , Estudos RetrospectivosRESUMO
The purpose of this study was to determine the utility of triple-phase helical computed tomography (CT) for differentiating canine hepatic masses. Seventy dogs with hepatic masses underwent triple-phase CT followed by surgical removal of the hepatic masses. Triple-phase helical CT scans for each dog included precontrast, arterial phase, portal venous phase, and delayed phase studies. The removed hepatic masses were histopathologically classified as hepatocellular carcinoma (n = 47), nodular hyperplasia (n = 14), and hepatic metastatic tumors (n = 9) in dogs. Of the 47 hepatocellular carcinomas, the most common CT findings included a heterogeneous pattern with hyper-, iso-, and hypoenhancement in both the arterial and portal venous phases (40/47, 85.1%). Of the 14 nodular hyperplasias, the most common CT findings were a homogeneous pattern with hyper- and isoenhancement in both the portal venous and delayed phases (13/14, 92.9%). Of nine hepatic metastatic tumors, the most common CT findings included a homogeneous hypoenhancement pattern in both the arterial and portal venous phases (8/9, 88.9%). In addition, 5 (55.6%) showed homogeneous hypoenhancement patterns in the delayed phase. Findings from our study indicated that triple-phase CT is a useful tool for preoperative differentiation of hepatocellular carcinoma, nodular hyperplasia, and hepatic metastatic tumors in dogs.
Assuntos
Doenças do Cão/diagnóstico por imagem , Hiperplasia/veterinária , Neoplasias Hepáticas/veterinária , Fígado/diagnóstico por imagem , Fígado/patologia , Animais , Carcinoma Hepatocelular/diagnóstico por imagem , Carcinoma Hepatocelular/veterinária , Cães , Feminino , Hiperplasia/diagnóstico por imagem , Neoplasias Hepáticas/diagnóstico por imagem , Masculino , Tomografia Computadorizada Espiral/veterináriaRESUMO
Background: Canine Legg Calvé Perthes disease (LCPD) occurs during the growth period, and the cause of ischemic necrosis of the femoral head during growth remains unclear. If LCPD-affected femoral head-derived mesenchymal stem cells (LCPD-MSCs) can be generated, they can be used as a new tool for the pathophysiological analysis of canine LCPD. Aim: To generate affected femoral head-derived mesenchymal stem cells (MSCs) from dogs with LCPD and investigate the mRNA expression levels of angiogenesis-related factors and osteogenic differentiation potency of LCPD-MSCs. Methods: This study was performed using affected femoral heads from dogs diagnosed with LCPD and underwent femoral head and neck ostectomy. The necrotic tissue was harvested from the LCPD-affected femoral head and cultured statically (LCPD group, n = 6). Canine bone marrow-derived MSCs (BM-MSCs) were used as controls (control group, n = 6). First, the morphology of the cultured cells was observed, and the expression of CD29, CD34, CD44, CD45, CD90, and major histocompatibility complex class II was analyzed using flow cytometry. Additionally, the trilineage differentiation potency of the LCPD-affected head-derived adherent cells was examined. Furthermore, the expression levels of HIF1A, VEGFA, VEGFB, and PDGFB mRNAs and the bone differentiation potency of LCPD-affected head-derived adherent cells were investigated. Results: LCPD-affected femoral head-derived adherent cells showed a fibroblast-like morphology, and the expression of cell surface antigens was similar to that of BM-MSCs. In addition, LCPD-affected femoral head-derived adherent cells showed the same trilineage differentiation potency as BM-MSCs and were consistent with MSC characteristics. Furthermore, the mRNA expression levels of angiogenesis-related factors could be objectively measured in LCPD-MSCs and those MSCs had bone differentiation potency. Conclusion: In the present study, canine LCPD-MSCs were successfully generated, suggesting their usefulness as a tool for pathological analysis of LCPD in dogs.
Assuntos
Doenças do Cão , Cabeça do Fêmur , Doença de Legg-Calve-Perthes , Células-Tronco Mesenquimais , Animais , Cães , Doença de Legg-Calve-Perthes/veterinária , Doença de Legg-Calve-Perthes/patologia , Células-Tronco Mesenquimais/fisiologia , Doenças do Cão/patologia , Cabeça do Fêmur/patologia , Diferenciação Celular , Osteogênese , Masculino , Células Cultivadas , FemininoRESUMO
Canine immune-mediated polyarthritis (IMPA) is an idiopathic disorder encompassing both erosive and non-erosive forms of rheumatoid arthritis (RA), with a clinical picture similar to human RA. Resemblance in major histocompatibility complex (MHC)-associated risk between the two was first noted within the specific amino acid motif known as the shared epitope (SE) on human leukocyte antigen DRB1. Following further identification of amino acids conferring risk for human RA outside the SE, this study was designed to examine amino acids both within and outside the classic SE in dachshunds, a breed with reported susceptibility to IMPA in Japan. Genome-wide association studies have linked positions 11, 13 and 71 with strong risk for human RA and important roles in antigen presentation to T cells. Sequence based genotyping of 16 case and 64 control dachshunds revealed strong associations comparable to human RA between IMPA risk and valine at position 11 (Val-11), phenylalanine at 13 (Phe-13), and arginine at 71 (Arg-71) on the dog leukocyte antigen (DLA)-DRB1 molecule (OR 2.89, 95%CI 1.3-6.4, p = 0.009), while association with the classic SE was significant only regarding homozygote frequency of the QRRAA haplotype-also carrying Val 11 and Phe 13 outside the SE (p = 0.04). Moreover, limited range in possible combinations of amino acids at positions 11, 13 and 71 starting with Val-11 among all DLA-DRB1 alleles registered with the GenBank and IPD-MHC canine databases, suggested potential of further single-breed analyses in dachshunds to clarify the disorder in terms of diagnosis, treatment, and epigenetic control, while clinical and immunopathogenetic similarities between human and dachshund RA also suggested the possibility of gaining insight into RA per se through study of canine IMPA as a spontaneous model of human RA.
Assuntos
Artrite Reumatoide , Doenças do Cão , Humanos , Cães , Animais , Epitopos/genética , Epitopos/química , Aminoácidos , Estudo de Associação Genômica Ampla/veterinária , Predisposição Genética para Doença , Artrite Reumatoide/genética , Artrite Reumatoide/veterinária , Cadeias HLA-DRB1/genética , Alelos , Doenças do Cão/genéticaRESUMO
MicroRNAs (miRNAs) are important regulators of intercellular signaling and are promising biomarkers in osteoarthritis (OA). In this study, comprehensive analysis was performed to identify miRNAs involved in the pathogenesis of spontaneous OA in dogs. Dogs diagnosed with OA based on radiography and arthroscopy of the stifle joint were included in the OA group. Dogs without any evidence of orthopedic disease were included in the unaffected group. To investigate miRNA expression levels, RNA sequencing analysis (RNA-seq) was performed in synovial tissue (OA group: n = 3, Unaffected group: n = 3) and RT-qPCR was performed in synovial tissue, synovial fluid and serum (OA group: n = 17, Unaffected group: n = 6), and compared between the two groups. The RNA-seq results showed that 57 miRNAs were significantly upregulated and 42 were significantly downregulated in the OA group. Specifically, miR-542 and miR-543 expression levels in the synovial tissue, synovial fluid, and serum were consistently higher in the OA group than in the unaffected group, suggesting that these miRNAs may be used as biomarkers for detecting canine OA. This is the first report to comprehensively analyze the expression patterns of miRNAs in the synovial tissue of dogs with spontaneous OA.
Assuntos
MicroRNAs , Osteoartrite , Cães , Animais , MicroRNAs/genética , MicroRNAs/metabolismo , Projetos Piloto , Osteoartrite/diagnóstico , Osteoartrite/genética , Osteoartrite/veterinária , Líquido Sinovial/metabolismo , Biomarcadores/metabolismoRESUMO
OBJECTIVE: The aim of this study was to determine the degree of bone deformities and hindlimb postural abnormalities in a standing position in awake Toy poodles with and without grade 2 medial patellar luxation (MPL) using high speed 320-row computed tomography (CT). METHODS: The limbs with grade 2 MPL (MPL-G2 group) and without any orthopaedic disorders (control group) were imaged in a standing position, without sedation or anaesthesia, using CT. In MPL-G2 group, images were obtained when the patella was luxated (G2-L group) and reduced (non-luxation, G2-NL group). Bone morphologies of the femur and tibia were quantified three-dimensionally. Hindlimb standing posture was evaluated by measuring femoral rotation and abduction angles, tibial rotation angle, metatarsal rotation angle, foot rotation angle, angle between the femoral anatomical axis and the mechanical axis of hindlimb and stifle joint line convergence angle. RESULTS: There were no significant differences in bone morphologic parameters between the MPL-G2 group (5 limbs) and the control group (6 limbs). In the G2-NL group, there were no significant hindlimb postural abnormalities. In contrast, in the G2-L group, significant hindlimb postural abnormalities including external rotation of femur, internal rotation of tibia and foot, external rotation of tarsal joint, large stifle joint convergence angle, genu varum and toe-in standing were observed. CONCLUSION: Dogs with grade 2 MPL have no bone deformities but show abnormal standing posture when the patella is luxated.
Assuntos
Doenças do Cão , Luxações Articulares , Luxação Patelar , Animais , Doenças do Cão/diagnóstico por imagem , Cães , Fêmur , Luxações Articulares/veterinária , Patela , Luxação Patelar/diagnóstico por imagem , Luxação Patelar/veterinária , Posição Ortostática , Joelho de Quadrúpedes/anatomia & histologia , Joelho de Quadrúpedes/diagnóstico por imagem , Tomografia Computadorizada por Raios X/veterinária , VigíliaRESUMO
Osteoarthritis (OA) is one of the major causes of chronic pain in dogs. However, the pathogenesis of OA has not been fully understood in dogs. The objective of this study was to comprehensively investigate the mRNA expression levels of proinflammatory cytokines, inflammatory mediators, nerve growth factor and its receptor, and matrix metalloproteinases in the synovium of dogs with spontaneous OA as well as to elucidate their relationships with the severity of synovitis. Dogs that were diagnosed with stifle OA on the basis of radiographic findings were included, and the degree of synovitis was observed using stifle arthroscopy. The dogs were assigned to two different groups depending on their synovitis scores: the low-grade group (score of 1 or 2; n = 8) and high-grade group (score of 3 to 5; n = 18). The dogs showing no evidence of orthopedic disease were included in the control group (n = 6). Synovial tissue samples were collected from the sites at which synovitis scores were assessed using arthroscopy. Total RNA was extracted from the collected synovial tissue, and cDNA was synthesized. Subsequently, RT-qPCR were performed using canine-specific primer sets for IL1B, IL6, CXCL8, TNF, TGFB1, PTGS2, PTGES, MMP3, MMP13, NGF, NTRK1, and PTGER4. Expression levels of IL1B, IL6, CXCL8, and MMP13 were significantly higher in the high-grade group than in the control group. In addition, expression levels of IL1B, CXCL8, TNF, and PTGS2 were significantly higher in the high-grade group than in the low-grade group. Expression levels of IL1B, IL6, CXCL8, TNF, PTGS2, and PTGER4 showed significant positive correlation with synovitis score. In conclusion, all mRNA expression levels in the synovial membrane varied according to the degree of synovitis in dogs with spontaneous OA. Thus, this study may partially elucidate the pathogenesis of synovitis in dogs with spontaneous OA.
Assuntos
Doenças do Cão/metabolismo , Regulação da Expressão Gênica , Osteoartrite/metabolismo , Osteoartrite/veterinária , Joelho de Quadrúpedes/metabolismo , Sinovite/metabolismo , Sinovite/veterinária , Animais , Doenças do Cão/patologia , Cães , Osteoartrite/patologia , Índice de Gravidade de Doença , Joelho de Quadrúpedes/patologia , Sinovite/patologiaRESUMO
OBJECTIVE: The aim of this study was to evaluate the bone deformities of the distal femur of Toy Poodles with medial patellar luxation (MPL) using computed tomography. METHODS: Computed tomography data from 37 stifles of Toy Poodles (16 normal, 10 grade 2 MPL and 11 grade 4 MPL) were analysed. For trochlear morphology, sulcus angle, medial or lateral trochlear inclination and trochlear angle were compared. For condylar morphology, distance between the apex of the medial or lateral trochlear ridge and the most caudal point of the medial or lateral femoral condyle (MC or LC), the length of the cranial/middle (anterior) part (MC(a) or LC(a')) and caudal (posterior) part (MC(p) or LC(p')) of distal femur and medial or lateral condyle width (MCW or LCW) were measured, and the ratios of medial-to-lateral condyles and anterior-posterior parts were compared. RESULTS: Sulcus angle was significantly higher, and medial trochlear inclination was significantly lower in grade 4 MPL group than other groups. Lateral trochlear inclination was not significantly different among 3 groups. Trochlear angles at proximal trochlear groove were significantly lower in grade 4 MPL group than other groups. MC/LC, MC(a)/LC(a'), MC(a)/MC(p), and LC(a')/LC(p') were significantly lower in grade 4 MPL group than other groups; however, there were no significant differences in MC(p)/LC(p') and MCW/LCW among three groups. CONCLUSION: Hypoplasia of the cranial and middle rather than caudal compartment of the distal femur was the primary morphological abnormality in Toy Poodles with grade 4 MPL.
Assuntos
Doenças do Cão , Luxação Patelar , Animais , Doenças do Cão/diagnóstico por imagem , Cães , Fêmur/diagnóstico por imagem , Articulação do Joelho , Luxação Patelar/diagnóstico por imagem , Luxação Patelar/veterinária , Joelho de Quadrúpedes , Tomografia Computadorizada por Raios X/veterináriaRESUMO
BACKGROUND: In recent years, surgical site infections caused by drug-resistant pathogens have emerged as a cause of concern in small animal practice. In this report, methicillin-resistant Staphylococcus aureus (MRSA) infections associated with tibial plateau leveling osteotomy (TPLO) is reported. However, there have been no reports on the treatment of MRSA infection following TPLO in dogs. This case report describes the use of a combination of vancomycin and rifampicin to treat MRSA infection following TPLO in a dog. CASE DESCRIPTION: A 7-year-old spayed female American cocker spaniel was referred for right hind limb lameness that did not improve with conservative treatment. The dog was diagnosed with cranial cruciate ligament rupture, for which TPLO was performed. Once the surgical wound was closed, the dog licked the skin on the surgical site, causing the injury to dehisce. MRSA was detected from the purulent discharge, and chloramphenicol was then administered based on the drug sensitivity test results. Because of the continued drainage, the implants were removed after the bone union of the osteotomy site was observed. Since this did not provide any relief to the existing condition, the antibiotic was changed to vancomycin at 132 days after TPLO surgery, and the infected location was cleaned many times through a drain tube placed into the tibia. However, the infection could not be controlled. Thus, a rifampicin and vancomycin combination was started. As a result, the purulent discharge disappeared and the fistula entirely closed on the 154th day after TPLO surgery. CONCLUSION: A combination of rifampicin and vancomycin may be effective for treating MRSA infection at the surgical site following TPLO surgery that does not heal despite implant removal and administration of vancomycin.
Assuntos
Doenças do Cão , Staphylococcus aureus Resistente à Meticilina , Animais , Ligamento Cruzado Anterior/cirurgia , Doenças do Cão/cirurgia , Cães , Feminino , Osteotomia/efeitos adversos , Osteotomia/veterinária , Rifampina/uso terapêutico , Joelho de Quadrúpedes/cirurgia , VancomicinaRESUMO
OBJECTIVE: The aim of this study was to investigate functional anatomy of the craniolateral bundle (CrLB) and caudomedial bundle (CdMB) of caudal cruciate ligament in normal Beagles. MATERIALS AND METHODS: Twelve stifle joints of 12 Beagle cadavers that were euthanatized for reasons other than musculoskeletal disease were used. Anatomical characteristics of femoral and tibial attachment regions of the caudal cruciate ligament were described, and location and area of the footprints of the CrLB and CdMB were objectively determined. The CrLB was artificially divided into two equal fibre bundle units and the CdMB was artificially divided into four equal bundle units. Changes in the fibre arrangement were recorded during stifle range of motion, and the tension of each fibre bundle unit at various stifle joint angles was objectively measured. RESULTS: The CrLB was attached to the intercondylar fossa of the femur, and the CdMB was attached to the medial surface of the medial condyle of the femur. The centre of the CrLB attachment site was located craniomedial to that of the CdMB on the tibia. During stifle range of motion, each divided fibre bundle unit in the CrLB and CdMB was twisted externally. The tension of entire CdMB tended to be higher than that of entire CrLB at 180 to 135 degrees of the stifle angles. In full extension angle, the tension of entire CdMB was 1.55 times higher than that of entire CrLB. CLINICAL SIGNIFICANCE: This study contributes to understanding of the biomechanics and the function of the caudal cruciate ligament, which help to elucidate the pathophysiology of caudal cruciate ligament tear and to develop stifle joint stabilization surgery.
Assuntos
Ligamento Cruzado Anterior , Joelho de Quadrúpedes , Animais , Fenômenos Biomecânicos , Cães , Fêmur , TíbiaRESUMO
We previously reported the non-viral derivation of transgene-free induced pluripotent stem cells (iPSCs) from somatic fibroblasts of a female beagle dog using an optimized induction medium and integration-free episomal vectors. Here, we report novel derivation of a male canine iPSC line OF35Y-iPS, which showed standard characteristics of pluripotency such as a strong gene expression profile of pluripotency markers, differentiation potential into all three germ layers, and normal karyotype (78XY). Furthermore, we demonstrated targeted integration of 2A-EGFP into the canine NANOS3 locus. The novel iPSC line would be a useful resource for stem cell research and regenerative veterinary medicine.
Assuntos
Células-Tronco Pluripotentes Induzidas , Animais , Diferenciação Celular , Cães , Feminino , Fibroblastos , Masculino , Pesquisa com Células-Tronco , TransgenesRESUMO
Induced pluripotent stem cells (iPSCs) are capable of providing an unlimited source of cells from all three germ layers and germ cells. The derivation and usage of iPSCs from various animal models may facilitate stem cell-based therapy, gene-modified animal production, and evolutionary studies assessing interspecies differences. However, there is a lack of species-wide methods for deriving iPSCs, in particular by means of non-viral and non-transgene-integrating (NTI) approaches. Here, we demonstrate the iPSC derivation from somatic fibroblasts of multiple mammalian species from three different taxonomic orders, including the common marmoset (Callithrix jacchus) in Primates, the dog (Canis lupus familiaris) in Carnivora, and the pig (Sus scrofa) in Cetartiodactyla, by combinatorial usage of chemical compounds and NTI episomal vectors. Interestingly, the fibroblasts temporarily acquired a neural stem cell-like state during the reprogramming. Collectively, our method, robustly applicable to various species, holds a great potential for facilitating stem cell-based research using various animals in Mammalia.
Assuntos
Fibroblastos/citologia , Células-Tronco Pluripotentes Induzidas/citologia , Células-Tronco Pluripotentes Induzidas/metabolismo , Mamíferos/metabolismo , Transgenes , Animais , Callithrix , Cães , Perfilação da Expressão Gênica , Vetores Genéticos/metabolismo , Camadas Germinativas/metabolismo , Células-Tronco Neurais/metabolismo , Plasmídeos/genética , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , RNA-Seq , Especificidade da Espécie , Suínos , VírusRESUMO
The objective of this paper was to evaluate the sequence of feline prepro-adrenomedullin (AM) and its tissue distribution and to investigate whether expression of feline AM mRNA increases in association with spontaneous cardiomyopathy. The feline prepro-AM cDNA sequence and deduced amino acids were 564 base pairs and 188 residues, respectively. The cDNA sequences of feline prepro-AM including AM and proadrenomedullin N-terminal 20 peptide showed high homology with those of other mammalian species. The mRNA expression of AM was detectable in various normal tissues. The mRNA levels of AM were elevated in hearts with cardiomyopathy compared with normal hearts. This study suggests that AM has an important role as a neurohumoral factor in cats with spontaneous heart diseases.
Assuntos
Cardiomiopatias/veterinária , Doenças do Gato/genética , Precursores de Proteínas/genética , RNA Mensageiro/genética , Sequência de Aminoácidos , Animais , Pareamento de Bases , Sequência de Bases , Cardiomiopatias/genética , Gatos , Clonagem Molecular , Primers do DNA , Humanos , Mamíferos/genética , Dados de Sequência Molecular , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
INTRODUCTION: Basic fibroblast growth factor (bFGF) is a promising cytokine in regenerative therapy for spinal cord injury. In this study, recombinant canine bFGF (rc-bFGF) was synthesized for clinical use in dogs, and the ability of rc-bFGF to differentiate canine bone marrow mesenchymal stem cells (BMSCs) into functional neurons was investigated. METHODS: The rc-bFGF was synthesized using a wheat germ cell-free protein synthesis system. The expression of rc-bFGF mRNA in the purification process was confirmed using a reverse transcription-polymerase chain reaction (RT-PCR). Western blotting was performed to confirm the antigenic property of the purified protein. To verify function of the purified protein, phosphorylation of extracellular signal-regulated kinase (ERK) was examined by in vitro assay using HEK293 cells. To compare the neuronal differentiation capacity of canine BMSCs in response to treatment with rc-bFGF, the cells were divided into the following four groups: control, undifferentiated, rh-bFGF, and rc-bFGF groups. After neuronal induction, the percentage of cells that had changed to a neuron-like morphology and the mRNA expression of neuronal markers were evaluated. Furthermore, to assess the function of the canine BMSCs after neuronal induction, changes in the intracellular Ca2+ concentrations after stimulation with KCl and l-glutamate were examined. RESULTS: The protein synthesized in this study was rc-bFGF and functioned as bFGF, from the results of RT-PCR, western blotting, and the expression of pERK in HEK293 cells. Canine BMSCs acquired a neuron-like morphology and expressed mRNAs of neuronal markers after neuronal induction in the rh-bFGF and the rc-bFGF groups. These results were more marked in the rc-bFGF group than in the other groups. Furthermore, an increase in intracellular Ca2+ concentrations was observed after the stimulation of KCl and l-glutamate in the rc-bFGF group, same as in the rh-bFGF group. CONCLUSIONS: A functional rc-bFGF was successfully synthesized, and rc-bFGF induced the differentiation of canine BMSCs into voltage- and glutamate-responsive neuron-like cells. Our purified rc-bFGF may contribute, on its own, or in combination with canine BMSCs, to regenerative therapy for spinal cord injury in dogs.
RESUMO
This prospective case study aimed to clarify the clinical significance of contrast-enhanced ultrasound (CEUS) for the differential diagnosis of canine adrenal tumors. Forty-three client-owned dogs with adrenal tumors were included. All dogs underwent CEUS, which was evaluated qualitatively and quantitatively. The peak signal intensity (PI), time to peak signal intensity (TPI), mean transit time (MTT), upslope, and downslope were calculated for each time-intensity curve. The histopathological diagnosis of each resected mass was compared with the CEUS findings and parameters. Enhancement distribution, vascularity, tortuous nourishing vessels, enhancement pattern, and late-phase enhancement did not differ significantly between adrenal cortical adenoma (CA), adenocarcinoma (CAC), and pheochromocytoma (PHEO) in qualitative assessment. In PHEO, the TPI was significantly more rapid compared with that in CA (P=0.0287) and CAC (P=0.0404). The MTT in PHEO was significantly shorter than that in CA (P=0.0016) and CAC (P=0.0003). Upslope in PHEO was larger than that in CAC (P=0.0406). Downslope in PHEO was significantly larger than that in CA (P=0.0048) and CAC (P=0.0018). A receiver operating characteristic curve analysis demonstrated that the area under the MTT curve yielded 0.91 for distinguishing PHEO from adrenocortical tumors in dogs; an MTT cut-off value less than 6,225 msec yielded a sensitivity of 69%, specificity of 94%, and likelihood ratio of 12.46. CEUS appears to be clinically applicable for the differential diagnosis between cortical and medullary origins of primary adrenal tumors in dogs.
Assuntos
Neoplasias das Glândulas Suprarrenais , Doenças do Cão , Feocromocitoma , Neoplasias das Glândulas Suprarrenais/diagnóstico por imagem , Neoplasias das Glândulas Suprarrenais/veterinária , Animais , Meios de Contraste , Diagnóstico Diferencial , Doenças do Cão/diagnóstico por imagem , Cães , Feocromocitoma/diagnóstico por imagem , Feocromocitoma/veterinária , Estudos Prospectivos , Ultrassonografia/veterináriaRESUMO
Osteoarthritis in cats is more prevalent because cats are living longer with advancement in veterinary medicine. Objective evaluation of behavioral changes in cats with osteoarthritis can facilitate an early diagnosis. The objective of this study was to investigate the utility of a novel activity monitor for analyzing physical activities and sleep quality in cats. First, a novel activity monitor (Plus Cycle®; JARMeC, Kanagawa, Japan), with a built-in three-directional accelerometer and an air pressure sensor specifically designed for cats, was compared to a human activity monitor (Actical®; Philips Respironics, OR, USA) previously used in cats (n = 10). Second, the validity of the measurement accuracy of the amount of physical activity, the number of vibrations, the number of jumps, and the resting and sleeping time was evaluated using Plus Cycle® in healthy cats (n = 6). Finally, the effects of gender and age of cats and time of day on the amount of physical activity, the number of vibrations, the number of jumps, and the resting and sleeping time were investigated in client-owned cats (n = 61). There were strong correlations between Plus Cycle® and Actical® in total activity (p < 0.05) and activity intensity (p < 0.05). When the physical activities were measured using Plus Cycle® in healthy cats, those data were quantified with high accuracy. In addition, it was also found to be very accurate in discriminating the resting and sleeping conditions of cats. In client-owned cats, there were no significant differences with respect to gender in any measured traits. The amount of physical activity and the number of jumps significantly decreased with the age of the cat. In contrast, the resting and sleeping times significantly increased with the age of the cat. In conclusion, Plus Cycle® can accurately and objectively assess physical activities and sleep quality with age of the cat, suggesting that this novel activity monitor can be used to manage the feline musculoskeletal health.
Assuntos
Monitores de Aptidão Física/veterinária , Monitorização Fisiológica/veterinária , Animais , Artrite/veterinária , Doenças do Gato , Gatos , Japão , Monitorização Fisiológica/instrumentação , Monitorização Fisiológica/métodos , Descanso , SonoRESUMO
The first case of feline true mycetoma because of a dermatophyte has been reported in this manuscript, although pseudomycetoma has been described in dogs and cats. The patient was a 9-year-old castrated male Persian cat weighing 4.2 kg with subcutaneous nodules on the dorsal trunk. Physical examination revealed two subcutaneous nodules (2.5 x 5.0 cm and 3.5 x 5.0 cm in size) that drained purulent exudates with cement-like substances containing yellowish granules. The impression smear of the yellowish granules demonstrated by PAS staining that they were masses of fungal septated hyphae (3-4 mum of width). From the nodular inflammation with fibrosis, fistulae draining from deep tissue and many grains containing abundant hyphal filaments, the case was diagnosed as mycetoma, complying with the definitive criteria of mycetoma. The etiologic fungus was molecularly as well as morphologically identified as Microsporum canis.
Assuntos
Doenças do Gato/microbiologia , Microsporum/isolamento & purificação , Micetoma/veterinária , Animais , Antifúngicos/uso terapêutico , Doenças do Gato/tratamento farmacológico , Gatos , DNA Fúngico/química , DNA Fúngico/genética , DNA Ribossômico/química , DNA Ribossômico/genética , Itraconazol/uso terapêutico , Masculino , Micetoma/tratamento farmacológico , Micetoma/microbiologia , RNA Ribossômico 28S/genética , Análise de Sequência de DNA , Pele/microbiologia , Pele/patologiaRESUMO
Twelve healthy Beagles were divided into 3 groups of 4 dogs each. To compare 2 methods of total hepatic vascular exclusion (THVE), we investigated the hemodynamic changes during THVE and assessed the influences on hepatic, renal and pancreatic biochemistry and the complications after THVE. In Group A, the thoracic aorta, hepatic pedicle and prehepatic and posthepatic caudal vena cava were occluded for 20 min, while in Group B, the celiac and cranial mesenteric arteries, hepatic pedicle and prehepatic and posthepatic caudal vena cava were occluded for 20 min. In Group C, a sham operation was performed. The arterial, venous and portal venous pressures and serum biochemistry parameters were measured before and during THVE and for 30 min after reperfusion. The carotid arterial pressure did not change in Group A during THVE, but decreased in Group B. The femoral and portal venous pressures in Group B increased significantly during THVE compared with those in Group C. With the exception of alanine aminotransferase, the serum biochemical profiles remained unchanged after the operation. For 7 days after the operation, no complications were observed in any of the dogs. In conclusion, occlusion of thoracic aorta, hepatic pedicle and prehepatic and posthepatic caudal vena cava is a feasible and safe method of hepatic vascular occlusion. This technique appears to be effective for canine hepatic surgery, such as removal of a large right-divisional hepatic tumor and attenuation of intrahepatic portosystemic shunt.