Development of molecular markers based on retrotransposons for the analysis of genetic variability in Moniliophthora perniciosa     

Mateus Ferreira Santana  Elza Fernandes de Araújo  Jorge Teodoro de Souza  Eduardo Seiti Gomide Mizubuti  Marisa Vieira de Queiroz 《European journal of plant pathology / European Foundation for Plant Pathology》2012,134(3):497-507

Moniliophthora perniciosa is a fungus that causes witches?? broom disease (WBD) in the cacao tree (Theobroma cacao). The M. perniciosa genome contains different transposable elements; this prompted an evaluation of the use of its retrotransposons as molecular markers for population studies. The inter-retrotransposon amplified polymorphism (IRAP) and retrotransposon-microsatellite amplified polymorphism (REMAP) techniques were used to study the variability of 70?M. perniciosa isolates from different geographic origins and biotypes. A total of 43 loci was amplified. Cluster analysis of different geographical regions of C biotype revealed two large groups in the state of Bahia, Brazil. Techniques using retrotransposon-based molecular markers showed advantages over previously used molecular techniques for the study of genetic variability in M. perniciosa.  相似文献   

Muscle reorganisation through local injection of stem cells in the diaphragm of mdx mice     

Thais Borges Lessa  Rafael Cardoso Carvalho  André Luis Rezende Franciolli  Lilian Jesus de Oliveira  RodrigoSilvadaNunes Barreto  David Feder  Fabiana Fernandes Bressan  Maria Angélica Miglino  Carlos Eduardo Ambrósio 《Acta veterinaria Scandinavica》2012,54(1):73

Background

The diaphragm is the major respiratory muscle affected by Duchenne muscular dystrophy (DMD) and is responsible for causing 80% of deaths. The use of mechanical forces that act on the body or intermittent pressure on the airways improves the quality of life of patients but does not prevent the progression of respiratory failure. Thus, diseases that require tissue repair, such as DMD, represent a group of pathologies that have great potential for cell therapy. The application of stem cells directly into the diaphragm instead of systemic application can reduce cell migration to other affected areas and increase the chances of muscle reorganisation. The mdx mouse is a suitable animal model for this research because its diaphragmatic phenotype is similar to human DMD. Therefore, the aim of this study was to assess the potential cell implantation in the diaphragm muscle after the xenotransplantation of stem cells.

Methods

A total of 9 mice, including 3 control BALB/Cmice, 3 5-month-old mdx mice without stem cell injections and 3 mdx mice injected with stem cells, were used. The animals injected with stem cells underwent laparoscopy so that stem cells from GFP-labelled rabbit olfactory epithelium could be locally injected into the diaphragm muscle. After 8 days, all animals were euthanised, and the diaphragm muscle was dissected and subjected to histological and immunohistochemical analyses.

Results

Both the fresh diaphragm tissue and immunohistochemical analyses showed immunopositive GFP labelling of some of the cells and immunonegativity of myoblast bundles. In the histological analysis, we observed a reduction in the inflammatory infiltrate as well as the presence of a few peripheral nuclei and myoblast bundles.

Conclusion

We were able to implant stem cells into the diaphragm via local injection, which promoted moderate muscle reorganisation. The presence of myoblast bundles cannot be attributed to stem cell incorporation because there was no immunopositive labelling in this structure. It is believed that the formation of the bundles may have been stimulated by cellular signalling mechanisms that have not yet been elucidated.  相似文献   

Comparative study of equine bone marrow and adipose tissue-derived mesenchymal stromal cells     

Ranera B  Ordovás L  Lyahyai J  Bernal ML  Fernandes F  Remacha AR  Romero A  Vázquez FJ  Osta R  Cons C  Varona L  Zaragoza P  Martín-Burriel I  Rodellar C 《Equine veterinary journal》2012,44(1):33-42

Reasons for performing study: Mesenchymal stromal cells (MSCs) represent an attractive source for regenerative medicine. However, prior to their application, fundamental questions regarding molecular characterisation, growth and differentiation of MSCs must be resolved. Objectives: To compare and better understand the behaviour of equine MSCs obtained from bone marrow (BM) and adipose tissue (AT) in culture. Methods: Five horses were included in this study. Proliferation rate was measured using MTT assay and cell viability; apoptosis, necrosis and late apoptosis and necrosis were evaluated by flow cytometry. The mRNA expression levels of 7 surface marker genes were quantified using RT‐qPCR and CD90 was also analysed by flow cytometry. Differentiation was evaluated using specific staining, measurement of alkaline phosphatase activity and analysis of the mRNA expression. Results: High interindividual differences were observed in proliferation in both cell types, particularly during the final days. Statistically significant differences in viability and early apoptosis of cultured AT‐ and BM‐MSCs were found. The highest values of early apoptosis were observed during the first days of culture, while the highest percentage of necrosis and late apoptosis and lowest viability was observed in the last days. Surface marker expression pattern observed is in accordance to other studies in horse and other species. Osteogenic differentiation was evident after 7 days, with an increasing of ALP activity and mRNA expression of osteogenic markers. Adipogenic differentiation was achieved in BM‐MSCs from 2 donors with one of the 16 media tested. Chondrogenic differentiation was also observed. Conclusions: Proliferation ability is different in AT‐MSCs and BM‐MSCs. Differences in viability and early apoptosis were observed between both sources and CD34 was only found in AT‐MSCs. Differences in their osteogenic and adipogenic potential were detected by staining and quantification of specific tissue markers. Potential relevance: To provide data to better understand AT‐MSCs and BM‐MSCs behaviour in vitro.  相似文献   

Evaluation of Nociception,Sedation, and Cardiorespiratory Effects of a Constant Rate Infusion of Xylazine Alone or in Combination with Lidocaine in Horses     

Juliano Ferreira Fernandes de Souza  Eduardo Raposo Monteiro  Daniela Campagnol  Renata Conti Ramos  Antonio Manoel Ferreira Frasson 《Journal of Equine Veterinary Science》2012

This study aimed to evaluate the effects of a constant rate infusion (CRI) of xylazine or xylazine in combination with lidocaine on nociception, sedation, and physiologic values in horses. Six horses were given intravenous (IV) administration of a loading dose (LD) of 0.55 mg/kg of xylazine followed by a CRI of 1.1 mg/kg/hr. The horses were randomly assigned to receive three treatments, on different occasions, administered 10 minutes after initiation of the xylazine CRI, as follows: control, physiologic saline; lidocaine low CRI (LLCRI), lidocaine (LD: 1.3 mg/kg, CRI: 0.025 mg/kg/min); and lidocaine high CRI (LHCRI), lidocaine (LD: 1.3 mg/kg, CRI: 0.05 mg/kg/min). A blinded observer assessed objective and subjective data for 50 minutes during the CRIs. In all treatments, heart and respiratory rates decreased, end-tidal carbon dioxide concentration increased, and moderate to intense sedation was observed, but no significant treatment effect was detected in these variables. Ataxia was significantly higher in LHCRI than in the control treatment at 20 minutes of infusion. Compared with baseline values, nociceptive threshold increased to as much as 79% in the control, 190% in LLCRI, and 158% in LHCRI. Nociceptive threshold was significantly higher in LLCRI (at 10 and 50 minutes) and in LHCRI (at 30 minutes) than in the control treatment. The combination of CRIs of lidocaine with xylazine produced greater increases in nociceptive threshold compared with xylazine alone. The effects of xylazine on sedation and cardiorespiratory variables were not enhanced by the coadministration of lidocaine. The potential to increase ataxia may contraindicate the clinical use of LHCRI, in combination with xylazine, in standing horses.  相似文献   

Virological Survey in free-ranging wildcats (Felis silvestris) and feral domestic cats in Portugal     

Duarte A  Fernandes M  Santos N  Tavares L 《Veterinary microbiology》2012,158(3-4):400-404

To determine the presence of viral pathogens in natural areas a survey was conducted on an opportunistic sample of fifty eight wild (Felis silvestris silvestris) and feral cats (F. s. catus). The biological materials included serum, lung tissue extract and stool. Feline leukemia virus p27 antigen was detected in 13/50 serum/lung tissue extract samples (26%), canine distemper virus antibodies were detected in 2/26 serum/lung tissue extract samples (7.7%), feline coronavirus RNA was present in 6/29 stool samples (20.7%) and feline parvovirus DNA in 2/29 stool samples (6.9%). Canine distemper virus RNA was not detected. Feline immunodeficiency virus and feline coronavirus antibodies were not detected. Evidence of exposure to feline leukemia virus, canine distemper virus, feline coronavirus and feline parvovirus was found in wild and feral cats raising the importance of performing a comprehensive survey to correctly evaluate the potential threat of infectious diseases to endangered species, namely to the wildcat and to the Iberian lynx, which is meant to be reintroduced after 2012 in Portugal.  相似文献   

Iridoid, phenylethanoid and flavonoid glycosides from Sideritis trojana     

Kirmizibekmez H  Ariburnu E  Masullo M  Festa M  Capasso A  Yesilada E  Piacente S 《Fitoterapia》2012,83(1):130-136

From the MeOH extract of Sideritis trojana, a new iridoid glycoside, 10-O-(E)-feruloylmelittoside (1) was obtained in addition to four known iridoid glycosides [melittoside (2), 10-O-(E)-p-coumaroylmelittoside (3), stachysosides E (4) and G (5)]. Moreover, five phenylethanoid glycosides [verbascoside (6), isoacteoside (7), lamalboside (8), leonoside A (9), isolavandulifolioside (10), three flavone glycosides (isoscutellarein 7-O-[6'-O-acetyl-β-allopyranosyl-(1→2)]-β-glucopyranoside (11), 4'-O-methyisoscutellarein 7-O-[6'-O-acetyl-β-allopyranosyl-(1→2)]-β-glucopyranoside (12), 3'-hydroxy-4'-O-methyisoscutellarein 7-O-[6'-O-acetyl-β-allopyranosyl-(1→2)]-β-glucopyranoside (13) and a benzylalcohol derivative (di-O-methylcrenatin) were obtained and identified. The structures were elucidated on the basis of NMR and HRMS data. All compounds were tested for their antioxidant activity by in vitro TEAC assay and some of them exhibited moderate activity (0.97-1.44 mM) when compared with the reference compound (quercetin 1.86 mM). Glycosides 6-13, the most active compounds in the TEAC assay, were also tested by flow cytometry to evaluate their ability to affect the levels of reactive oxygen species (ROS) in human prostate cancer cells (PC3).  相似文献   

Evaluation in broilers of the probiotic properties of Pichia pastoris and a recombinant P. pastoris containing the Clostridium perfringens alpha toxin gene     

Gil de los Santos JR  Storch OB  Fernandes CG  Gil-Turnes C 《Veterinary microbiology》2012,156(3-4):448-451

The probiotic properties of Pichia pastoris and of a recombinant P. pastoris containing the Clostridium perfringens alpha toxin gene were evaluated in broilers. One-day-old chicks randomly divided in four groups were fed with commercial feed devoid of antibacterials. The control group (1) received plain food, while the other groups were supplemented with either P. pastoris (2), the recombinant P. pastoris (3) or Bacillus cereus var. Toyoi (4). At day 49, live weights, feed efficiency and seroconversions were higher (P<0.05) in the supplemented groups than in the control groups. Group 3 showed the best results, while group 2 had lower weight gain than groups 3 and 4 although food conversion was better than in group 4. Seroconversions were not different (P>0.05) among the supplemented groups. Adverse reactions were not observed in histopathologic evaluation. We concluded that P. pastoris and the recombinant P. pastoris could be used as probiotics in broilers.  相似文献   

Non-accidental injuries found in necropsies of domestic cats: a review of 191 cases     

de Siqueira A  Cassiano FC  Landi MF  Marlet EF  Maiorka PC 《Journal of Feline Medicine and Surgery》2012,14(10):723-728

Animal cruelty is defined as a deliberate action that causes pain and suffering to an animal. In Brazil, legislation known as the Environmental Crimes Law states that cruelty toward all animal species is criminal in nature. From 644 domestic cats necropsied between January 1998 and December 2009, 191 (29.66%) presented lesions highly suggestive of animal cruelty. The main necroscopic finding was exogenous carbamate poisoning (75.39%) followed by blunt-force trauma (21.99%). Cats from 7 months to 2 years of age were the most affected (50.79%). In Brazil, violence is a public health problem and there is a high prevalence of domestic violence. Therefore, even if laws provide for animal welfare and protection, animals are common targets for violent acts. Within a context of social violence, cruelty toward animals is an important parameter to be considered, and the non-accidental lesions that were found are evidence of malicious actions.  相似文献   

Constraints using the liquid fraction from roadside grass as a bio-based fertilizer     

Hannah Scott  Marcella Fernandes de Souza  Erik Meers  Naomi Laura Jane Rintoul-Hynes 《植物养料与土壤学杂志》2023,186(4):387-396

Background

Roadside grass cuttings are currently considered a waste product due to their association with road sweepings as contaminated waste, therefore, their potential as a biofertilizer is understudied.

Aim

This study aimed to determine whether grass liquid fraction (GLF) collected from a roadside verge in Maldegem, Belgium, and pressed using a screw press was suitable as a biofertilizer.

Methods

The characterization of the heavy metal content of the GLF was conducted using an ICP-OES. From May to September 2019, a pot experiment was set up using a randomized block design to compare tomato plant growth, yield, and nutrition for GLF-treated plants to two commercial fertilizers and tap water as a control.

Results

The heavy metal content of the GLF was below the maximum permissible concentrations (MPCs) for organic fertilizers as set out by the European Comission fertilizer regulation 1069/2009 and 1107/2009 (European Comission, 2019). However, despite having a fairly well-balanced nutrient content (0.1% N, 0.04% P₂O₅, and 0.2% K₂O), GLF had a negative effect on the growth, root weight, and yield of the tomato plants, killing six out of ten plants. GLF also promoted mold growth in the soil of some plants. Since the GLF was uncontaminated, heavy metal toxicity did not cause the negative effect.

Conclusions

Previous research showed that liquid fractions from some plants negatively affect the growth of others due to allelopathic chemicals; this, together with the stimulation of fungal growth, could have caused the negative effects observed. Future experiments will investigate the herbicidal property of GLF and possible treatments to potentially recover the nutrients contained within the GLF for application as a biofertilizer.  相似文献   

Serum vascular endothelial growth factor in dogs with soft tissue sarcomas     

G. Fernandes de Queiroz  M. Lúcia Zaidan Dagli  S. Aparecida Meira  J.  Maria Matera 《Veterinary and comparative oncology》2013,11(3):230-235

This work aimed to evaluate serum vascular endothelial growth factor (VEGF) in 25 dogs with soft tissue sarcoma, and in 30 healthy dogs. Blood was collected once time from the control animals and three times, in the same way, from animals with sarcoma. Blood count was performed in the blood collected, and serum VEGF was measured by enzyme‐linked immunosorbent assay quantitative method. Serum VEGF in control animals was similar to patients with soft tissue sarcoma. There was a reduction in serum VEGF after the sarcoma resection. There was positive correlation between serum VEGF and neutrophil counts, and negative between VEGF and hemoglobin content in animals with sarcoma. Animals with hemangiopericytoma showed higher serum VEGF levels compared to the patients with malignant peripheral nerve sheath. Circulating blood cells can contribute to elevate VEGF serum concentrations in dogs with soft tissue sarcomas and a possible role of VEGF in the angiogenesis of these tumors.  相似文献