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1.
Front Vet Sci ; 11: 1388152, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38711537

RESUMO

Thyroglossal duct cyst represents a congenital anomaly of the cervical region, rarely documented in animals. Although previously reported in dogs, cats, horses, goats, pigs, and calves, never in birds. This report describes a rare case of thyroglossal duct cyst in a hen. A necropsy of a Transylvanian Naked Neck hen carried following diphtheroid mucocutaneous lesions. The necropsy revealed a large, cyst-like structure measuring 0.5 cm at the level of the caudal edge of the left thyroid gland. Histologically, the cystic mass, bordered by 1-2 lines of well-differentiated ciliated cuboidal cells, presented nuclear immunoreactivity for Thyroid transcription factor 1. To the best of the authors' knowledge, there are no previous records of thyroglossal duct cysts in avians. Moreover, this is the first case describing a thyroglossal duct cyst in a hen.

2.
Front Vet Sci ; 10: 1213150, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-37396991

RESUMO

Caprine papillomaviruses (ChPVs, Capra hircus papillomaviruses) were detected and quantified for the first time using droplet digital polymerase chain reaction (ddPCR) in blood samples of 374 clinically healthy goats from farms located in Italy, Romania, and Serbia. Overall, ddPCR revealed ChPV DNA in 78 of the 374 examined samples, indicating that ~21% of the goats harbored circulating papillomavirus DNA. In particular, in Italian goat farms, ChPV genotypes were detected and quantified in 58 of 157 blood samples (~37%), 11 of 117 samples from Serbian farms (~9.4%), and 9 of 100 from Romanian blood samples (9%). Blood samples from Italian goat farms showed a high prevalence of ChPV1, which was detected in 45 samples (28.6%). The ChPV2 genotype was detected in 13 samples (~8.3%). Therefore, significant differences in prevalence and genotype distributions were observed. On Serbian and Romanian farms, no significant differences were observed in the genotype prevalence of ChPVs. Molecular findings are consistent with ChPV prevalence, characterized by a territorial distribution similar to that of papillomaviruses in other mammalian species. Furthermore, this study showed that ddPCR is a very sensitive and accurate assay for ChPV detection and quantification. The ddPCR may be the molecular diagnostic tool of choice, ultimately providing useful insights into the molecular epidemiology and field surveillance of ChPV.

3.
Transbound Emerg Dis ; 69(6): 3850-3857, 2022 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-36335589

RESUMO

Ovine papillomavirus (OaPV) comprises four genotypes; OaPV1, OaPV2 and OaPV4 are fibropapillomaviruses within the genus Deltapapillomavirus, whereas OaPV3 is an epitheliotropic virus that belongs to the genus Dyokappapapillomavirus. To date, all of them have been known to infect sheep only. OaPV1, OaPV2 and OaPV4 have been associated with ovine cutaneous and mucosal fibropapillomas, whereas OaPV3 is a key factor in the squamous cell carcinoma pathway of the sheep skin. Whole blood samples obtained from 128 cattle at public slaughterhouses were investigated using droplet digital polymerase chain reaction (ddPCR). ddPCR is a new-generation PCR technique that enables an accurate and absolute quantification of target molecules with high sensitivity and specificity. All OaPVs were detected by identification and quantification of nucleic acids using specific fluorescent probes. Of 128 blood samples, 100 (∼78%) showed OaPV infections. Further, 42, 35 and 23 blood samples showed single, double and triple OaPV infections, respectively. OaPV1 was responsible for 22 single infections, OaPV2 caused 16 single infections and OaPV3 and OaPV4 caused two single infections each. OaPV1 and OaPV2 were the most frequent ovine viruses in dual and triple infections. In many blood samples, both ovine deltapapillomavirus and dyokappapapillomavirus were found to be transcriptionally active, as shown by the detection and quantification of E5 oncogene transcripts for OaPV1, L1 transcripts for OaPV2, E6 and E7 transcripts for OaPV3 and E6 for OaPV4. OaPVs were found in the blood samples from cattle that shared grasslands rich in bracken ferns known to contain immunosuppressant substances. Furthermore, OaPVs were also found in cattle from intensive livestock farming without any contact with sheep. Because OaPV DNA was detected in both grass hay and corn silage, it is conceivable that these feed may be the viral sources.


Assuntos
Doenças dos Bovinos , Deltapapillomavirus , Infecções por Papillomavirus , Doenças dos Ovinos , Ovinos , Animais , Bovinos , Deltapapillomavirus/genética , Papillomaviridae/genética , Pele/patologia , Reação em Cadeia da Polimerase/veterinária , Reação em Cadeia da Polimerase/métodos , Técnicas de Amplificação de Ácido Nucleico/veterinária , Infecções por Papillomavirus/epidemiologia , Infecções por Papillomavirus/veterinária , Doenças dos Ovinos/epidemiologia , Doenças dos Ovinos/diagnóstico
4.
Animals (Basel) ; 12(20)2022 Oct 20.
Artigo em Inglês | MEDLINE | ID: mdl-36290249

RESUMO

Obesity is a common feature in horses suffering from metabolic syndrome. While adipokines involvement as biomarkers is better established in human pathology, little data are available on horses. This study aimed to investigate the possible association and relationship between selected metabolic parameters and morphological traits in equine metabolic syndrome. Adiposity was evaluated using body condition score (BCS) and cresty neck score (CNS). Plasma levels of total cholesterol, insulin, NEFA, and adipokines (omentin and chemerin) were determined using enzyme-linked immunosorbent assays. Spearman correlation, univariate linear regression analysis and hierarchical clustering were performed. Significant positive correlations were observed between NEFA and bodyweight (r = 0.322; p = 0.006), BCS (r = 0.295; p = 0.013), and CNS (r = 0.267; p = 0.024), total cholesterol and bodyweight (r = 0.262; p = 0.027), and omentin and CNS (r = 0.234; p = 0.049). Cluster analysis supported these results and provided more details on the relationships between studied variables within and between the four resulting groups. These findings highlight NEFA, chemerin, and omentin as valuable biomarkers that could be further analyzed in other horse breeds for a better understanding of equine metabolic pathology.

5.
Pharmaceutics ; 14(7)2022 Jul 14.
Artigo em Inglês | MEDLINE | ID: mdl-35890358

RESUMO

Helicobacter pylori is the first formally recognized bacterial carcinogen and the most important single digestive pathogen responsible for the induction of gastroduodenal diseases such as gastritis, peptic ulcer, and, finally, gastric neoplasia. The recently reported high rates of antimicrobial drug resistance hamper the current therapies of H. pylori, with therapeutic failure reaching up to 40% of patients. In this context, new treatment options and strategies are urgently needed, but the successful development of these new therapeutic tools is conditioned by the understanding of the high adaptability of H. pylori to the gastric acidic environment and the complex pathogenic mechanism. Due to several advantages, including good antibacterial efficiency, possible targeted delivery, and long tissular persistence, silver nanoparticles (AgNPs) offer the opportunity of exploring new strategies to improve the H. pylori therapy. A new paradigm in the therapy of H. pylori gastric infections using AgNPs has the potential to overcome the current medical limitations imposed by the H. pylori drug resistance, which is reported for most of the current organic antibiotics employed in the classical therapies. This manuscript provides an extensive overview of the pathology of H. pylori-induced gastritis, gastric cancer, and extradigestive diseases and highlights the possible benefits and limitations of employing AgNPs in the therapeutic strategies against H. pylori infections.

6.
Animals (Basel) ; 12(6)2022 Mar 19.
Artigo em Inglês | MEDLINE | ID: mdl-35327178

RESUMO

African swine fever has a significant economic, social and environmental impact due to official regulation of the disease, namely the mass depopulation of all pigs in confirmed outbreaks. The main objective of the present study was to statistically analyze the possible correlation between the number of outbreaks and infected pigs from backyard farms with the altitude, seasonal average annual humidity, and average temperature during spring, summer, autumn and winter, as well as the distance from forests, rivers, and lakes in Romania. The study included all infected backyard pigs with African swine fever virus (n = 7764) and outbreaks (n = 404) that occurred in Romania between 6 February 2020 and 2 March 2021. The number of infected pigs and ASF outbreaks were significantly higher in localities at altitudes between 100 and 500 m, average annual humidity between 60% and 80%, average spring temperature between 10 and 14 °C, average summer temperature between 16 and 22 °C, average autumn temperature between 10 and 13 °C, average winter temperatures between -1 and 2 °C, and at distances of less than 5 km from the forests, less than 15 km from lakes and less than 5 km from the rivers. The number of affected pigs decreased significantly at summer temperatures below 16 °C.

7.
Front Vet Sci ; 8: 790392, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34881323

RESUMO

Ovine papillomaviruses (OaPVs) were detected and quantified, for the first time, using droplet digital polymerase chain reaction (ddPCR) and real-time quantitative PCR (qPCR) via blood samples of 165 clinically healthy sheep. OaPV DNA was detected in 126 blood samples (~76.4%). DdPCR detected OaPV DNA in 124 samples; in only two additional samples positive for real-time qPCR, ddPCR failed to detect the presence of any OaPVs. In 70 of the positive samples (~55.6%), a single OaPV infection was observed, 12 of which were caused by OaPV1 (~17.1%) and 14 by OaPV2 (20%). OaPV3 was responsible for 19 single infections (~27.1%), and OaPV4 for 25 single infections (~35.7%). Multiple OaPV coinfections were observed in 56 (~44.4%) positive samples. OaPV coinfections caused by two genotypes were observed in 31 positive samples (~55.4%), with dual OaPV3/OaPV4 infection being the most prevalent as seen in 11 blood samples. In addition, five OaPV1/OaPV4, four OaPV1/OaPV2, four OaPV2/OaPV3, four OaPV1/OaPV3, and three OaPV2/OaPV4 dual coinfections were also detected. OaPV coinfections by triple and quadruple genotypes were detected in 24 (~42.8%) and only one (~1.8%) of coinfected blood samples, respectively. Multiple infections caused by OaPV1/OaPV3/OaPV4 genotypes were the most prevalent, as observed in 12 (50%) blood samples harboring triple OaPV infections. This study showed that ddPCR is the most sensitive and accurate assay for OaPV detection and quantification thus outperforming real-time qPCR in terms of sensitivity and specificity. Therefore, ddPCR may represent the molecular diagnostic tool of choice, ultimately providing useful insights into OaPV molecular epidemiology and field surveillance.

8.
Life (Basel) ; 11(12)2021 Dec 16.
Artigo em Inglês | MEDLINE | ID: mdl-34947937

RESUMO

Obesity has become a serious health problem with frequent occurrence both in human and animal populations. It is estimated that it may affect over 85% of the human population and 70-80% of horses and cows by 2030. Fat cow syndrome (FCS) is a combination of metabolic, digestive, infectious, and reproductive disorders that affects obese periparturient dairy cows, and occurs most frequently in loose-housing systems, where periparturient and dry cows are fed and managed in one group disregarding the lactation stages. Equine metabolic syndrome (EMS) was named after human metabolic syndrome (MetS) and has insulin dysregulation as a central and consistent feature. It is often associated with obesity, although EMS may occur in a lean phenotype as well. Other inconsistent features of EMS are cardiovascular changes and adipose dysregulation. Laminitis is the main clinical consequence of EMS. MetS holds a 30-years old lead in research and represents a clustering of risk factors that comprise abdominal obesity, dyslipidemia, hypertension, and hyperglycemia (impaired fasting glucose or type 2 diabetes mellitus-T2DM), which are associated with doubled atherosclerotic cardiovascular disease risk, and a 5-fold increased risk for T2DM. The main aim of this review is to provide critical information for better understanding of the underlying mechanisms of obesity-related metabolic dysfunction in animals, especially in cows and horses, in comparison with MetS. Human medicine studies can offer suitable candidate mechanisms to fill the existing gap in the literature, which might be indispensable for owners to tackle FCS, EMS, and their consequences.

9.
Vet Sci ; 8(12)2021 Nov 26.
Artigo em Inglês | MEDLINE | ID: mdl-34941817

RESUMO

The World Organisation for Animal Health has listed African swine fever as the most important deadly disease in domestic swine around the world. The virus was recently brought from South-East Africa to Georgia in 2007, and it has since expanded to Russia, Eastern Europe, China, and Southeast Asia, having a devastating impact on the global swine industry and economy. In this study, we report for the first time the molecular characterization of nine African swine fever virus (ASFV) isolates obtained from domestic pigs in Mures County, Romania. All nine Romanian samples clustered within p72 genotype II and showed 100% identity with all compared isolates from Georgia, Armenia, Russia, Azerbaijan, Ukraine, Belarus, Lithuania, and Poland. This is the first report of ASFV genotype II in the country.

10.
Med Pharm Rep ; 94(1): 106-111, 2021 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-33629057

RESUMO

INTRODUCTION: In recent years a link between periodontitis and cardiovascular diseases has been proposed. Oxidative stress has been found to be involved in their pathophysiology. OBJECTIVES: The aim of this study was to demonstrate that a wire ligature placed around the first mandibular molars of the rat causes inflammation in the periodontium and lipid deposits in the aorta, via oxidative stress. METHODS: Twenty male Wistar rats were divided randomly into two groups of 10: Control group and Periodontitis group. An orthodontic wire ligature was placed around the cervix of the first mandibular molar and was kept in place for 4 weeks. Clinical attachment loss was determined to assess the gingival recession. Aorta was harvested after 4 weeks and the malondialdehyde (MDA) levels in the aorta tissue homogenate were determined. Lipid deposits were evaluated using Oil Red staining and immunofluorescence under confocal microscopy. Total lipid deposits in the aorta were assessed using a computer morphometry system. RESULTS: Significant gingival recession was observed in the Periodontitis group in comparison with the Control group. Increased MDA levels were present in the aorta in the ligatured rats in comparison with the Control group. Significant lipid deposits were observed in the aorta of the Periodontitis group as opposed to the Control group. CONCLUSIONS: The affixing of an orthodontic wire ligature produces gingival recession, increased oxidative stress and lipid deposits in the rat descending aorta.

11.
Pathogens ; 9(8)2020 Jul 29.
Artigo em Inglês | MEDLINE | ID: mdl-32751272

RESUMO

Prohibitin 2 (PHB2), an inner mitochondrial membrane (IMM) protein, has recently been identified as a novel receptor involved in parkin-mediated mitophagy. In the field of veterinary medicine, the role of PHB2 in parkin-mediated mitophagy was described, for the first time, in urothelial cells of cattle, naturally infected with bovine papillomavirus (BPV). The BPV2 and BPV13 E5 oncoprotein, responsible for abortive infections in urothelial cells, was detected by RT-PCR. Severe ultrastructural abnormalities of the inner mitochondrial membrane were detected using transmission electron microscopy. PHB2 formed a functional complex with PHB1. PHB2 was significantly overexpressed in mitochondrial fractions from urothelial mucosa samples taken from cattle harbouring BPV infection. PHB2 overexpression could be attributed to mitochondrial dysfunction, as its expression levels in the cytosolic, microsomal, and nuclear fractions were seen to be unmodified. Immunoprecipitation studies revealed the interaction between PHB2 and phosphorylated forms of both PINK1 and parkin. Furthermore, PHB2 interacted with LC3-II, a marker of autophagosomal membranes and autophagy receptors, such as p62 and optineurin. PHB2 was shown to interact with transcription factor EB (TFEB), which is activated following parkin-mediated mitophagy, and embryonic stem cell-expressed Ras (ERAS), a constitutive protein coded by ERas. Western blot analysis revealed a significant overexpression of unphosphorylated TFEB in mitochondrial and nuclear fractions from urothelial mucosa samples from cattle suffering from BPV infection. Finally, PHB2 interacted with ERAS, believed to be involved in mitophagosome maturation. Taken together, the molecular and ultrastructural findings of this study suggested that BPV infection is responsible for parkin-dependent mitophagy, in the pathway of which PHB2 plays a crucial role.

12.
BMC Vet Res ; 16(1): 296, 2020 Aug 17.
Artigo em Inglês | MEDLINE | ID: mdl-32807166

RESUMO

BACKGROUND: Ovine pulmonary adenocarcinoma (OPA) is a neoplastic disease caused by exogenous Jaagsiekte Sheep Retrovirus (exJSRV). The prevalence of JSRV-related OPA in Eastern European countries, including Romania is unknown. We aimed to investigate: the prevalence and morphological features of OPA (classical and atypical forms) in the Transylvania region (Romania), the immunophenotype of the pulmonary tumors and their relationships with exJSRV infection. A total of 2693 adult ewes slaughtered between 2017 and 2019 in two private slaughterhouses from Transylvania region (Romania) was evaluated. Lung tumors were subsequently assessed by cytology, histology, immunocytochemistry, immunohistochemistry, electron microscopy and DNA testing. RESULTS: Out of 2693 examined sheep, 34 had OPA (1.26% prevalence). The diaphragmatic lobes were the most affected. Grossly, the classical OPA was identified in 88.24% of investigated cases and the atypical OPA in 11.76% that included solitary myxomatous nodules. Histopathology results confirmed the presence of OPA in all suspected cases, which were classified into acinar and papillary types. Myxoid growths (MGs) were diagnosed in 6 classical OPA cases and in 2 cases of atypical form. Lung adenocarcinoma was positive for MCK and TTF-1, and MGs showed immunoreaction for Vimentin, Desmin and SMA; Ki67 expression of classical OPA was higher than atypical OPA and MGs. JSRV-MA was identified by IHC (94.11%) in both epithelial and mesenchymal cells of OPA. Immunocytochemistry and electron microscopy also confirmed the JSRV within the neoplastic cells. ExJSRV was identified by PCR in 97.05% of analyzed samples. Phylogenetic analysis revealed the presence of the exJSRV type 2 (MT809678.1) in Romanian sheep affected by lung cancer and showed a high similarity with the UK strain (AF105220.1). CONCLUSIONS: In this study, we confirmed for the first time in Romania the presence of exJSRV in naturally occurring OPA in sheep. Additionally, we described the first report of atypical OPA in Romania, and to the best of our knowledge, in Eastern Europe. Finally, we showed that MGs have a myofibroblastic origin.


Assuntos
Adenocarcinoma de Pulmão/veterinária , Retrovirus Jaagsiekte de Ovinos/isolamento & purificação , Neoplasias Pulmonares/veterinária , Adenomatose Pulmonar Ovina/epidemiologia , Adenocarcinoma de Pulmão/patologia , Adenocarcinoma de Pulmão/virologia , Animais , Feminino , Imuno-Histoquímica/veterinária , Imunofenotipagem/veterinária , Retrovirus Jaagsiekte de Ovinos/classificação , Retrovirus Jaagsiekte de Ovinos/ultraestrutura , Neoplasias Pulmonares/patologia , Neoplasias Pulmonares/virologia , Microscopia Eletrônica/veterinária , Filogenia , Prevalência , Romênia/epidemiologia , Ovinos , Carneiro Doméstico
13.
J Med Life ; 13(4): 580-586, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-33456610

RESUMO

The associations between periodontitis and cardiovascular diseases have been intensely studied in recent years. Oxidative stress is involved in the initiation and both progression of periodontitis and atherosclerosis. Antioxidants can reduce the effects of oxidative stress on inflammatory diseases. Our aim was to measure the effects of a grape seed extract (GSE), rich in antioxidants, on atherosclerosis caused by ligature-induced periodontitis in rats. Thirty male Wistar rats were randomly divided into three groups of 10: control group, periodontitis group, and periodontitis group treated with GSE (GSE group). Periodontitis was induced by placing an orthodontic wire around the cervix of the first mandibular molar and keeping it in place for 4 weeks. On days 1, 7 and 28, blood samples were taken to assess oxidative stress and inflammation markers (malondialdehyde and glutathione - MDA, reduced glutathione - GSH, C reactive protein) and lipids. After 4 weeks, the animals were euthanized, and aortas were collected for histopathologic examination. MDA was significantly higher in Periodontitis group compared to the other groups only at day 7. GSH was significantly increased in the Control and GSE groups on days 1 and 7, compared to Periodontitis group and on day 28 higher in GSE vs. Periodontitis groups. C reactive protein was significantly increased in the Periodontitis group on days 1 and 7 compared to both groups. Cholesterol was significantly decreased in the aortas of GSE group at day 28 compared to the Periodontitis group. Oral administration of a grape seed extract reduces the oxidative stress, inflammation and atherosclerosis in a rat model of ligature-induced periodontitis.


Assuntos
Aterosclerose/tratamento farmacológico , Extrato de Sementes de Uva/uso terapêutico , Periodontite/complicações , Periodontite/etiologia , Animais , Antioxidantes/farmacologia , Aorta/patologia , Aterosclerose/sangue , Proteína C-Reativa/metabolismo , HDL-Colesterol/sangue , Glutationa/metabolismo , Extrato de Sementes de Uva/administração & dosagem , Ligadura , Masculino , Malondialdeído/sangue , Periodontite/sangue , Periodontite/patologia , Ratos Wistar
14.
Vet Microbiol ; 236: 108396, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31500722

RESUMO

Autophagy is a powerful tool that host cells use to defend against viral infection. Mitophagy, the selective autophagic removal of dysfunctional mitochondria was upregulated in urothelial cancer cells harbouring bovine papillomavirus (BPV) infection, as detected by the expression of BPV E5 protein, the major oncoprotein of bovine Deltapapillomavirus genus. HIF-1α-induced mitophagy receptors, BNIP3 and BNIP3L/Nix, were found to be overexpressed in these cells. The BNIP3 and BNIP3L/Nix receptors were amplified, and amplicon sequencing showed homology between bovine BNPI3 and BNIP3L/Nix sequences deposited in GenBank (accession number: NM_001076366.1 and NM_001034614.2, respectively). The transcripts and protein levels of BNIP3 and BNIP3L/Nix were significantly overexpressed in hypoxic neoplastic cells relative to healthy, non-neoplastic cells. BNIP3 and BNIP3L/Nix interacted with the LC3 protein, a marker of autophagosome (mitophagosome) membrane, ERAS, a small GTPase, and p62, known to be a specific autophagy receptor protein, that plays a role in mitochondrial priming for mitophagy and subsequent elimination. ERAS also interacted with the BPV E5 oncoprotein at mitochondrial level. Furthermore, in anti-Bag3 mitochondrial immunoprecipitates, a complex composed of the Hsc70/Hsp70 chaperone, CHIP co-chaperone, Synpo2, ERAS, LC3, p62, BNPI3, and BNIP3L/Nix was also detected. Bag3 may play a role in mitophagosome formation together with the Synpo2 protein and may be involved in the degradation of Hsc70/Hsp70-bound CHIP-ubiquitinated cargo, in association with its chaperone. ERAS may be involved in mitophagosome maturation via the PI3K signalling pathway. Ultrastructural findings revealed the presence of mitochondria exhibiting severe fragmentation and loss of cristae, as well as numerous mitochondria-containing autophagosomes.


Assuntos
Papillomavirus Bovino 1 , Papillomavirus Bovino 4 , Doenças dos Bovinos/virologia , Infecções por Papillomavirus/veterinária , Proteínas Proto-Oncogênicas/metabolismo , Urotélio/citologia , Animais , Bovinos , Doenças dos Bovinos/patologia , Masculino , Proteínas de Membrana , Proteína Oncogênica p21(ras)/genética , Proteína Oncogênica p21(ras)/metabolismo , Proteínas Oncogênicas Virais , Infecções por Papillomavirus/virologia , Proteínas Supressoras de Tumor/genética , Proteínas Supressoras de Tumor/metabolismo , Bexiga Urinária/metabolismo , Neoplasias da Bexiga Urinária/metabolismo , Neoplasias da Bexiga Urinária/ultraestrutura , Neoplasias da Bexiga Urinária/veterinária , Neoplasias da Bexiga Urinária/virologia , Urotélio/metabolismo
15.
Vet Microbiol ; 234: 51-60, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31213272

RESUMO

E5 protein, the major oncoprotein of the bovine Deltapapillomavirus genus, has been detected in 17 of the 19 urothelial cancers by molecular and morphological procedures. In 10 urothelial cancers, the oxygen sensitive subunit HIF-1α, which is upregulated by hypoxia, was overexpressed. Mitophagy, the selective autophagic removal of dysfunctional mitochondria, was upregulated in hypoxic neoplastic cells infected by BPVs which was mediated by FUNDC1, a mitochondrial outer-membrane protein. The FUNDC1 receptor was amplified by PCR, and amplicon sequencing showed a 100% homology with bovine FUNDC1 sequences deposited in GenBank (accession number: NM_001104982). Both transcripts and protein levels of FUNDC1 were significantly decreased in hypoxic neoplastic cells relative to healthy, non-neoplastic cells. FUNDC1 interacted with the LC3 protein, a marker of autophagosome (mitophagosome) membrane, the Hsc70/Hsp70 chaperone, and Bag3 co-chaperone. Bag3 may play a role in mitophagosome formation together with the Synpo2 protein, and may be involved in the degradation of Hsc70/Hsp70-bound CHIP-ubiquitinated cargoes, in association with its chaperone. Ultrastructural findings revealed the presence of mitochondria exhibiting severe fragmentation and loss of cristae, as well as numerous mitochondria-containing autophagosomes. Total and phosphorylated GTPase dynamin-related protein 1 (DRP1), which plays a crucial role in mitochondrial fission, a pre-requisite for mitophagy, was overexpressed at the mitochondrial level. Total and phosphorylated mitochondrial fission factor (Mff), mitochondrial fission protein 1 (Fis1), mitochondrial dynamics 51 (MiD51), and MiD49, which are DRP1 receptors responsible and/or co-responsible for its mitochondrial recruitment were overexpressed.


Assuntos
Deltapapillomavirus/patogenicidade , Mitocôndrias/patologia , Proteínas Mitocondriais/metabolismo , Mitofagia , Urotélio/virologia , Animais , Bovinos , Feminino , Proteínas de Ligação ao GTP/genética , Hipóxia , Microscopia Eletrônica de Transmissão , Mitocôndrias/ultraestrutura , Mitocôndrias/virologia , Proteínas Mitocondriais/genética , Proteínas Oncogênicas Virais/genética , Fosforilação , Urotélio/citologia , Urotélio/patologia
16.
Vet Microbiol ; 229: 147-152, 2019 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-30642591

RESUMO

The feline leukemia virus subgroup C receptors (FLVCRs) were originally cloned as virus receptors, but are now believed to function also as heme transporters and are expressed in a broad range of tissues in a wide range of mammalian species. Expression of FLVCR1 and FLVCR2 was investigated in 19 bovine papillomavirus-associated urinary bladder cancers and in 15 non-neoplastic samples of bladder from cattle. E5 oncoprotein of bovine Deltapapillomaviruses (δPVs) was detected in 17 of the 19 bladder cancers. Flvcr1 and Flvcr2 were amplified and sequenced both in neoplastic and non-neoplastic samples showing a 100% identity with bovine Flvcr1 and Flvcr2 mRNA sequences present in GenBank database (accession numbers: NM_001206019.1 and NM_001192143.1, respectively). Reverse transcription (RT)-PCR showed that Flvcr1 and Flvcr2 were overexpressed in 4 and 5 out of 19 urothelial cancers, respectively, but in none of the non-neoplastic samples. In addition, western blot analysis detected higher levels of FLVCR1 and FLVCR2 in samples in which transcripts were not increased, suggesting post-translational changes to these proteins. Increased FLCVR1 and FLVCR2 was also observed immunohistochemically in the neoplastic cells. Immunolabeling for FLVCR1 was seen in the cytoplasm and plasm membrane of urothelial cancer cells, wheras immunolabeling for FLVCR2 was present within the nucleus. This is the first time that FLVCR expression has been investigated in bovine tissues and the first to suggest that expression could be increased in cancers. Additional studies are required to define the role, if any, of FLVCR in papillomavirus-associated cancer cells.


Assuntos
Papillomavirus Bovino 1 , Doenças dos Bovinos/virologia , Proteínas de Membrana Transportadoras/metabolismo , Receptores Virais/metabolismo , Neoplasias da Bexiga Urinária/veterinária , Urotélio/metabolismo , Animais , Bovinos , Regulação Neoplásica da Expressão Gênica , Proteínas de Membrana Transportadoras/genética , Infecções por Papillomavirus/metabolismo , Infecções por Papillomavirus/veterinária , Infecções por Papillomavirus/virologia , Receptores Virais/genética , Neoplasias da Bexiga Urinária/patologia , Neoplasias da Bexiga Urinária/virologia
17.
J Med Life ; 12(4): 342-353, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-32025252

RESUMO

The restoration of voluntary muscle activity in posttraumatic paraplegia in both animal experiments and other clinical applications requires reproducibility of a technically-demanding microsurgical procedure, limited by physicians' understanding of Brunelli's spinal cord grafting paradigm. The insufficient clinical investigation of the long-term benefits of the CNS-PNS graft application warrants additional inquiry. The objective of this study is to explore the potential benefits of the first replicated, graft-induced neuroregeneration of denervated skeletal muscle regarding long-term clinical outcomes and to investigate the effect of Cerebrolysin on neuromodulation. A randomized study evaluating 30 rats, approved by the National Animal Ethics Advisory Committee was performed. The medication was administered postoperatively. For 14 days, 12 rats received Cerebrolysin (serum), 11 received NaCl 0.9% (shams), and 7 were controls. For microsurgery, the lateral corticospinal tract T10 was grafted to the denervated internal obliquus abdominal muscle. On day 90, intraoperative proof of reinnervation was observed. On day 100, 15 rats were euthanized for fixation, organ removal, and extensive histology-morphology examination, and the Wei-Lachin statistical procedure was employed. After an open revision of 16 rats, 8 were CMAP positive. After intravenous Vecuronium application, two (Cerebrolysin, NaCl) out of two rats showed an incomplete compound muscle action potential (CMAP) loss due to glutamatergic and cholinergic co-transmission, while two others showed a complete loss of amplitude. Cerebrolysin medication initiated larger restored muscle fiber diameters and less scarring. FB+ neurons were not observed in the brain but were observed in the Rexed laminae. Brunelli's concept was successfully replicated, demonstrating the first graft induced existence of cholinergic and glutamatergic neurotransmission in denervated grafted muscles. Statistics of the histometric count of muscle fibers revealed larger fiber diameters after Cerebrolysin. Brunelli's CNS-PNS experimental concept is suitable to analyze graft-neuroplasticity focused on the voluntary restoration of denervated skeletal muscles in spinal cord injury. Neuroprotection by Cerebrolysin is demonstrated.


Assuntos
Sistema Nervoso Central/fisiologia , Músculo Esquelético/inervação , Regeneração Nervosa/fisiologia , Sistema Nervoso Periférico/fisiologia , Potenciais de Ação/efeitos dos fármacos , Aminoácidos/farmacologia , Animais , Sistema Nervoso Central/efeitos dos fármacos , Feminino , Músculo Esquelético/efeitos dos fármacos , Regeneração Nervosa/efeitos dos fármacos , Sistema Nervoso Periférico/efeitos dos fármacos , Ratos Sprague-Dawley , Reprodutibilidade dos Testes
18.
Med Pharm Rep ; 92(Suppl No 3): S39-S44, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31989107

RESUMO

INTRODUCTION: In recent years a link between periodontitis and cardiovascular diseases has been proposed. Oxidative stress has been found to be involved in their pathophysiology. OBJECTIVES: The aim of this study was to demonstrate that a wire ligature placed around the first mandibular molars of the rat causes inflammation in the periodontium and lipid deposits in the aorta, via oxidative stress. METHODS: Twenty male Wistar rats were divided randomly into two groups of 10: Control group and Periodontitis group. An orthodontic wire ligature was placed around the cervix of the first mandibular molar and was kept in place for 4 weeks. Clinical attachment loss was determined to assess the gingival recession. Aorta was harvested after 4 weeks and the malondialdehyde (MDA) levels in the aorta tissue homogenate were determined. Lipid deposits were evaluated using Oil Red staining and immunofluorescence under confocal microscopy. Total lipid deposits in the aorta were assessed using a computer morphometry system. RESULTS: Significant gingival recession was observed in the Periodontitis group in comparison with the Control group. Increased MDA levels were present in the aorta in the ligatured rats in comparison with the Control group. Significant lipid deposits were observed in the aorta of the Periodontitis group as opposed to the Control group. CONCLUSIONS: The affixing of an orthodontic wire ligature produces gingival recession, increased oxidative stress and lipid deposits in the rat descending aorta.

20.
Colloids Surf B Biointerfaces ; 163: 155-166, 2018 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-29291501

RESUMO

Wound healing is a complex and dynamic process that involves modifying the wound environment depending on the patient's health status. Scar formation depends on many factors that influence wound healing, are important to bear in mind because most of the negative factors involved can be stopped by implementing an adequate treatment. The consensus in wound therapy recommends dressings that should keep a moist and alkaline environment thus creating a protective barrier against mechanical stress and secondary infections, in view of promoting granulation. In the current paper, we aimed to synthesize a polymer-based sponge containing chitosan-sodium hyaluronate-resveratrol (CHR) and to evaluate its regenerative potential. The process of synthesizing the CHR polymer was described before microtomography analysis was conducted and the density and porosity of the obtained sponges was assessed. The cytotoxicity was evaluated in vitro. By undertaking the in vivo testing of the CHR polymer, we aimed to determine the CHR sponge's potential to stimulate tissue regeneration after inflicting a controlled, reproducible and measurable skin wound in an animal model. Skin punch biopsies were harvested from the healed area and were subjected to histopathological evaluation. The results obtained in this study confirmed that this polymer accelerates the formation of granulation facilitating wound healing, while also achieving a bacteriostatic outcome.


Assuntos
Bandagens , Quitosana/química , Ácido Hialurônico/química , Pele/efeitos dos fármacos , Estilbenos/farmacologia , Ferida Cirúrgica/tratamento farmacológico , Cicatrização/efeitos dos fármacos , Animais , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Avaliação Pré-Clínica de Medicamentos , Fibroblastos/citologia , Fibroblastos/efeitos dos fármacos , Humanos , Masculino , Camundongos , Camundongos Endogâmicos ICR , Porosidade , Resveratrol , Pele/lesões , Estilbenos/química , Ferida Cirúrgica/patologia
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