Association of thoracic spine deformity and cardiovascular disease in a mouse model for Marfan syndrome.

AIMS: Cardiovascular manifestations are a major cause of mortality in Marfan syndrome (MFS). Animal models that mimic the syndrome and its clinical variability are instrumental for understanding the genesis and risk factors for cardiovascular disease in MFS. This study used morphological and ultrastructural analysis to the understanding of the development of cardiovascular phenotypes of the the mgΔloxPneo model for MFS. METHODS AND RESULTS: We studied 6-month-old female mice of the 129/Sv background, 6 wild type (WT) and 24 heterozygous animals from the mgΔloxPneo model. Descending thoracic aortic aneurysm and/or dissection (dTAAD) were identified in 75% of the MFS animals, defining two subgroups: MFS with (MFS+) and without (MFS-) dTAAD. Both subgroups showed increased fragmentation of elastic fibers, predominance of type I collagen surrounding the elastic fiber and fragmentation of interlaminar fibers when compared to WT. However, only MFS animals with spine tortuosity developed aortic aneurysm/dissection. The aorta of MFS+ animals were more tortuous compared to those of MFS- and WT mice, possibly causing perturbations of the luminal blood flow. This was evidenced by the detection of diminished aorta-blood flow in MFS+. Accordingly, only MFS+ animals presented a process of concentric cardiac hypertrophy and a significantly decreased ratio of left and right ventricle lumen area. CONCLUSIONS: We show that mgΔloxPneo model mimics the vascular disease observed in MFS patients. Furthermore, the study indicates role of thoracic spine deformity in the development of aorta diseases. We suggest that degradation of support structures of the aortic wall; deficiency in the sustenance of the thoracic vertebrae; and their compression over the adjacent aorta resulting in disturbed blood flow is a triad of factors involved in the genesis of dissection/aneurysm of thoracic aorta.

Use of polyclonal antibodies, electron microscopy and histopathology to detect iridovirus-like particles in bullfrogs / Uso de anticorpos policlonais, microscopia eletrônica e histologia para detecção de partículas semelhantes ao iridovírus em rãs-touro

Iridoviruses of the Ranavirus genus have been implicated in the decline in amphibians worldwide, capable of affecting animals both in the wild and in captivity. This study aimed to detect iridovirus-like particles from three frog farms in southeastern Brazil using primary polyclonal antibodies, transmission electron microscopy (TEM) and histologic findings. The target organs were liver and kidneys. Sixty adults and sixty tadpoles of bullfrogs (Lithobates catesbeianus) were used in the study. TEM revealed the presence of iridovirus-like particles in hepatic tissue using the negative staining technique. Positive results were also observed by immunoelectron microscopy and immunocytochemistry (ICC). The histological analysis of the samples showed liver hemorrhage and corpuscles inclusion in hepatocytes as well as glomerulotubular degeneration and necrosis in the kidneys. The methods used in this study were highly efficient to detect the presence of iridovirus-like particles and possible infection of ranavirus.

Os iridovirus do gênero Ranavirus têm sido implicados no declínio dos anfíbios em todo o mundo afetando animais de vida livre e aqueles em cativeiro. O objetivo deste estudo foi detectar a presença de partículas semelhantes ao iridovírus em três ranários na região sudeste do Brasil, utilizando anticorpos policlonais primários, microscopia eletrônica de transmissão (MET) e achados histológicos. Os órgãos alvo foram o fígado e os rins. Sessenta rãs-touro adultas (Lithobates catesbeianus) e sessenta girinos da mesma espécie foram usados para o estudo. A MET revelou a presença de partículas semelhantes ao iridovírus em tecido hepático utilizando a técnica de contrastação negativa. Os resultados positivos foram também observados por imunomicroscopia eletrônica e imunocitoquímica. As análises histológicas nas mesmas amostras evidenciaram hemorragia no fígado e corpúsculos de inclusão em hepatócitos, e degeneração glomerulotubular e necrose nos rins. Os métodos usados neste estudo foram altamente eficientes na detecção de partículas semelhantes ao iridovírus e possivel infecção por ranavirus.

Use of polyclonal antibodies, electron microscopy and histopathology to detect iridovirus-like particles in bullfrogs / Uso de anticorpos policlonais, microscopia eletrônica e histologia para detecção de partículas semelhantes ao iridovírus em rãs-touro

Iridoviruses of the Ranavirus genus have been implicated in the decline in amphibians worldwide, capable of affecting animals both in the wild and in captivity. This study aimed to detect iridovirus-like particles from three frog farms in southeastern Brazil using primary polyclonal antibodies, transmission electron microscopy (TEM) and histologic findings. The target organs were liver and kidneys. Sixty adults and sixty tadpoles of bullfrogs (Lithobates catesbeianus) were used in the study. TEM revealed the presence of iridovirus-like particles in hepatic tissue using the negative staining technique. Positive results were also observed by immunoelectron microscopy and immunocytochemistry (ICC). The histological analysis of the samples showed liver hemorrhage and corpuscles inclusion in hepatocytes as well as glomerulotubular degeneration and necrosis in the kidneys. The methods used in this study were highly efficient to detect the presence of iridovirus-like particles and possible infection of ranavirus.(AU)

Os iridovirus do gênero Ranavirus têm sido implicados no declínio dos anfíbios em todo o mundo afetando animais de vida livre e aqueles em cativeiro. O objetivo deste estudo foi detectar a presença de partículas semelhantes ao iridovírus em três ranários na região sudeste do Brasil, utilizando anticorpos policlonais primários, microscopia eletrônica de transmissão (MET) e achados histológicos. Os órgãos alvo foram o fígado e os rins. Sessenta rãs-touro adultas (Lithobates catesbeianus) e sessenta girinos da mesma espécie foram usados para o estudo. A MET revelou a presença de partículas semelhantes ao iridovírus em tecido hepático utilizando a técnica de contrastação negativa. Os resultados positivos foram também observados por imunomicroscopia eletrônica e imunocitoquímica. As análises histológicas nas mesmas amostras evidenciaram hemorragia no fígado e corpúsculos de inclusão em hepatócitos, e degeneração glomerulotubular e necrose nos rins. Os métodos usados neste estudo foram altamente eficientes na detecção de partículas semelhantes ao iridovírus e possivel infecção por ranavirus.(AU)

PVALB diminishes [Ca2+] and alters mitochondrial features in follicular thyroid carcinoma cells through AKT/GSK3ß pathway.

We have identified previously a panel of markers (C1orf24, ITM1 and PVALB) that can help to discriminate benign from malignant thyroid lesions. C1orf24 and ITM1 are specifically helpful for detecting a wide range of thyroid carcinomas, and PVALB is particularly valuable for detecting the benign Hürthle cell adenoma. Although these markers may ultimately help patient care, the current understanding of their biological functions remains largely unknown. In this article, we investigated whether PVALB is critical for the acquisition of Hürthle cell features and explored the molecular mechanism underlying the phenotypic changes. Through ectopic expression of PVALB in thyroid carcinoma cell lines (FTC-133 and WRO), we demonstrated that PVALB sequesters free cytoplasmic Ca(2+), which ultimately lowers calcium levels and precludes endoplasmic reticulum (ER) Ca(2+) refilling. These results were accompanied by induced expression of PERK, an ER stress marker. Additionally, forced expression of PVALB reduces Ca(2+) inflow in the mitochondria, which can in turn cause changes in mitochondria morphology, increase mitochondria number and alter subcellular localization. These findings share striking similarity to those observed in Hürthle cell tumors. Moreover, PVALB inhibits cell growth and induces cell death, most likely through the AKT/GSK-3ß. Finally, PVALB expression coincides with Ca(2+) deposits in HCA tissues. Our data support the hypothesis that the loss of PVALB plays a role in the pathogenesis of thyroid tumors.

Isolation of chicken astrovirus from specific pathogen-free chicken embryonated eggs.

Astroviruses have been associated with enteric disorders in many animal species, including chickens. Here, we describe the isolation, propagation, and pathological characteristics of chicken astrovirus (CAstV) in specific pathogen free (SPF) chicken embryonated eggs (CEE) from chickens with diarrhea and runting-stunting syndrome. The CEE were inoculated via the yolk sac route. Viral confirmation was carried out using PCR techniques and transmission electron microscopy negative staining with ammonium molybdate. The intestinal contents were screened for CAstV, and differential diagnostic testing was performed for avian nephritis virus, avian rotavirus, avian reovirus, chicken parvovirus, infectious bronchitis virus, and fowl adenovirus Group I to detect co-infection with other infectious agents. Seven- or 14-day-old CEEs presented with hemorrhages, edema, a gelatinous aspect, deformities, and dwarfism. The supporting membranes did not show any alterations. Here, we have described the isolation of CAstV and its pathological characteristics in SPF CEE.

Occurrence of ferret enteric coronavirus in Brazil (Preliminary Report) / Ocorrência de coronavírus entérico de ferrets no Brasil - Nota Prévia

Ferret enteric coronavirus (FECV) is associated to the epizootic catarrhal enteritis (ECE) in ferrets (Mustela putorius furo). In this study, we report the occurrence of this agent in four diarrheic stool samples of domestic ferrets, analyzed by negative staining transmission electron microscopy and a specific RT-PCR assay targeting the nucleocapsid (N) gene. These findings are the first report of FECV in Brazil and address the importance of this virus on the etiology of enteric disorders in ferrets.

Coronavírus entérico de furões (FECV) é associado à enterite catarral epizoótica (ECE) em furões (Mustela putorius furo). Neste estudo, relatamos a ocorrência deste agente em quatro amostras fecais diarreicas de furões domésticos, analisadas por microscopia eletrônica de transmissão (contrastação negativa) e RT-PCR específica e direcionada ao gene de nucleocapsídeo (N). Estes achados constituem o primeiro relato de FECV no Brasil e remetem para a importância deste vírus na etiologia de quadros entéricos nestes animais.

Occurrence of ferret enteric coronavirus in Brazil (Preliminary Report) / Ocorrência de coronavírus entérico de ferrets no Brasil - Nota Prévia

Ferret enteric coronavirus (FECV) is associated to the epizootic catarrhal enteritis (ECE) in ferrets (Mustela putorius furo). In this study, we report the occurrence of this agent in four diarrheic stool samples of domestic ferrets, analyzed by negative staining transmission electron microscopy and a specific RT-PCR assay targeting the nucleocapsid (N) gene. These findings are the first report of FECV in Brazil and address the importance of this virus on the etiology of enteric disorders in ferrets.(AU)

Coronavírus entérico de furões (FECV) é associado à enterite catarral epizoótica (ECE) em furões (Mustela putorius furo). Neste estudo, relatamos a ocorrência deste agente em quatro amostras fecais diarreicas de furões domésticos, analisadas por microscopia eletrônica de transmissão (contrastação negativa) e RT-PCR específica e direcionada ao gene de nucleocapsídeo (N). Estes achados constituem o primeiro relato de FECV no Brasil e remetem para a importância deste vírus na etiologia de quadros entéricos nestes animais.(AU)