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71.
Gupta R Yarnall BW Giuliano EA Kanwar JR Buss DG Mohan RR 《Veterinary ophthalmology》2011,14(5):304-312
Objective To evaluate the safety and efficacy of mitomycin C (MMC) in prevention of canine corneal scarring. Methods With an in vitro approach using healthy canine corneas, cultures of primary canine corneal fibroblasts or myofibroblasts were generated. Primary canine corneal fibroblasts were obtained by growing corneal buttons in minimal essential medium supplemented with 10% fetal bovine serum. Canine corneal myofibroblasts were produced by growing cultures in serum‐free medium containing transforming growth factor β1 (1 ng/mL). Trypan blue assay and phase‐contrast microscopy were used to evaluate the toxicity of three doses of MMC (0.002%, 0.02% and 0.04%). Real‐time PCR, immunoblot, and immunocytochemistry techniques were used to determine MMC efficacy to inhibit markers of canine corneal scarring. Results A single 2‐min treatment of 0.02% or less MMC did not alter canine corneal fibroblast or keratocyte phenotype, viability, or growth. The 0.02% dose substantially reduced myofibroblast formation (up to 67%; P < 0.001), as measured by the change in RNA and protein expression of fibrosis biomarkers (α‐smooth muscle actin and F‐actin). Conclusion This in vitro study suggests that a single 2‐min 0.02% MMC treatment to the canine corneal keratocytes is safe and may be useful in decreasing canine corneal fibrous metaplasia. In vivo studies are warranted. 相似文献
72.
Across the southeastern United States, changing disturbance regimes threaten the integrity of pine flatwoods, reducing their suitability as habitat for a variety of avian species, many of which are considered of conservation priority. Prescribed burning and roller chopping are management practices widely used in pine flatwoods to control shrubs and increase the growth of herbaceous vegetation. However, the impacts these treatments have on this system and its associated avian communities are largely unknown. We examined the effects of prescribed burning and roller chopping on pine flatwoods avian communities using a paired sample approach. Avian communities were compared between treated (e.g., burned) and untreated areas. We considered abundance and richness of all birds, but also groups of conservation concern, and those likely impacted by seasonal management practices (e.g., breeding and non-breeding categories, and woodland, successional-scrub, and grassland guilds). Dormant season burning resulted in decreases in avian species richness and abundance within the non-breeding, overwintering community. In situations where the maintenance or enhancement of this avian group is desired, growing season burning appears beneficial and should be used in preference to dormant season burning. Growing season burning benefited breeding avian communities, resulting in increases in species richness and abundance within this group. Dormant season roller chopping had no effect on species richness or abundance of any avian category or guild. Increases in abundance within breeding avian communities were observed following growing season roller chopping and a roller chopping/burning combination. Grazing, did not mediate the effects of prescribed burning or roller chopping on avian species richness or abundance. The use of dormant season burning is likely inappropriate in situations where conservation and management of non-breeding, overwintering species is a priority. The use of growing season roller chopping appears to provide a potential treatment alternative in situations where positive effects on breeding species are desired. Overall, maintenance of avian abundance and richness may be best achieved through the diverse application of prescribed burning and roller chopping based on season, frequency, and space. 相似文献
73.
Danielli Giuliano Perez Carmem Silvia Fontanetti 《Water, air, and soil pollution》2011,218(1-4):437-444
The destination of sewage sludge is a problem faced by sewage treatment plants (STPs). Many alternatives have been sought, such as the application of sewage sludge in degraded soils and in agriculture as fertilizer. However, due to the risk of contamination with pathogens and/or metals, the use of sludge should be done cautiously. By the habits that diplopods present, they have been considered good environmental indicators for soil analysis. In this study, animals from the Rhinocricus padbergi species were exposed to two sewage sludge samples from two STPs in the São Paulo State, for different periods. The midgut of the animals were removed and histologically processed and subjected to histochemical tests. It was detected the following tissular responses: clusters of haemocytes through the cells of the fat body layer, increase in the quantity of intracellular granules in the cells of the fat body layer, increase in the release of secretion vesicles of the intestinal epithelium, and intense vacuolization of the cytoplasm of epithelial cells. The results suggest the presence of toxic substances to the studied species in both sludge samples used. 相似文献
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Laura D. Barnes Jacqueline W. Pearce Linda M. Berent Derek B. Fox Elizabeth A. Giuliano 《Veterinary ophthalmology》2010,13(4):251-258
A 27 kg, 6‐year‐old, male castrated German shorthaired pointer presented to the University of Missouri, Veterinary Teaching Hospital with the complaint of progressive exophthalmia of 2 years duration optical density (OD). Lack of retropulsion OD was noted on physical examination. Anterior segment examination OU and fundic examination OS did not reveal any abnormalities. Examination of the fundus OD revealed focal scleral indentation of the inferior nasal globe. The indentation changed location with globe movement OD. MRI and CT scan revealed a well‐circumscribed, approximately 2 cm in diameter mass located caudal and ventral to the affected globe that appeared to communicate with the nictitating membrane with absence of any bony involvement. A modified lateral orbitotomy was recommended and performed to remove the orbital mass and nictitating membrane en‐bloc. Histopathology and immunohistochemistry of the mass confirmed a diagnosis of nodular granulomatous episcleritis (NGE). Postoperatively, the dog developed absolute keratoconjunctivitis sicca (KCS). Examples of primary episcleral inflammation in the dog include diffuse episcleritis, NGE, nodular fasciitis, fibrous histiocytoma, proliferative conjunctivitis/keratoconjunctivitis, pseudotumor, and Collie granuloma. The etiology of these episcleral inflammations is presumed to be immune mediated. To our knowledge, this is the first report of NGE affecting the orbital region of a dog. Development of absolute KCS resulting from excision of the nictitating membrane is also supported by this case. 相似文献
78.
Dylan G. Buss Ajay Sharma Elizabeth A. Giuliano Rajiv R. Mohan 《Veterinary ophthalmology》2010,13(4):211-218
Objective Mitomycin C (MMC) is used clinically to treat corneal scarring in human patients. We investigated the safety and efficacy of MMC to treat corneal scarring in horses by examining its effects at the early and late stages of disease using an in vitro model. Procedure An in vitro model of equine corneal fibroblast (ECF) developed was used. The ECF or myofibroblast cultures were produced by growing primary ECF in the presence or absence of transforming growth factor beta‐1 (TGFβ1) under serum‐free conditions. The MMC dose for the equine cornea was defined with dose‐dependent trypan blue exclusion and (3‐4,5‐dimethylthiazol‐2‐yl)‐2,5‐diphenyltetrazolium bromide (MTT) assays after applying MMC to the cultures once for 2 min. The efficacy of MMC to control corneal scarring in horses was determined by measuring mRNA and protein expression of corneal scarring markers (alpha‐smooth muscle actin and F‐actin) with western blotting, immunocytochemistry and/or quantitative real‐time polymerase chain reactions. Results A single 2‐min treatment of 0.02% or less MMC did not alter ECF phenotype, viability, or cellular proliferation whereas 0.05% or higher MMC doses showed mild‐to‐moderate cellular toxicity. The TGFβ1 at 1 ng/mL showed significant myofibroblast formation in ECF under serum‐free conditions. A single 2‐min, 0.02% MMC treatment 24 h (early) after TGFβ1 stimulation significantly reduced conversion of ECF to myofibroblasts, however, a single 0.02% MMC treatment 11 days after TGFβ1 stimulation showed moderate myofibroblast inhibition. Conclusions That MMC safely and effectively reduced scarring in ECF by reducing the degree of transdifferentiation of corneal fibroblasts to myofibroblasts in vitro. Further clinical in vivo investigations are warranted using MMC in horses. 相似文献
79.
Dylan G. Buss Elizabeth Giuliano Ajay Sharma Rajiv R. Mohan 《Veterinary ophthalmology》2010,13(5):301-306
Objective To determine if hybrid adeno‐associated virus serotype 2/5 (AAV5) vector can effectively deliver foreign genes into the equine cornea without causing adverse side effects. The aims of this study were to: (i) evaluate efficacy of AAV5 to deliver therapeutic genes into equine corneal fibroblasts (ECFs) using enhanced green fluorescent protein (EGFP) marker gene, and (ii) establish the safety of AAV5 vector for equine corneal gene therapy. Material Primary ECF cultures were harvested from healthy donor equine corneas. Cultures were maintained at 37°C in humidified atmosphere with 5% CO2. Procedure AAV5 vector expressing EGFP under control of hybrid cytomegalovirus + chicken β‐actin promoter was applied topically to ECF. Expression of delivered EGFP gene in ECF was quantified using fluorescent microscopy. Using fluorescent staining, the total number of cells and transduction efficiency of tested AAV vector was determined. Phase contrast microscopy, trypan blue and TUNEL assays were used to determine toxicity and safety of AAV5 for ECFs. Results Topical AAV5 application successfully transduced significant numbers of ECFs. Transduction efficiency was 13.1%. Tested AAV5 vector did not cause phenotype change or significant cell death and cell viability was maintained. Conclusions Tested AAV5 vector is effective and safe for gene therapy in ECFs in vitro. 相似文献
80.
Giovanni Manarolla Giuliano Pisoni Giuseppe Sironi Tiziana Rampin 《Veterinary microbiology》2010,140(1-2):1-8
Fifteen strains of Avipoxvirus from different avian species were isolated and molecular biologically characterized. Most strains did not produce evident pocks on the chorioallantoic membranes of commercial and specific-pathogen free embryonated chicken eggs where, on the contrary, microscopic signs of viral growth were always detected. Polymerase chain reaction of highly conserved P4b gene was positive for all cases confirming to be a reliable diagnostic method for Avipoxvirus. Sequencing of these amplicons confirmed most strains clustered either with Fowlpox virus or with Canarypox virus whereas a possible new clade could be hypothesized for one strain from Japanese quail. Classification of Avipoxvirus strains by amplification of the newly identified locus fpv140 revealed major limitations as only five samples were positive. These results underline the importance to undertake similar studies on higher numbers of Avipoxvirus isolates and on wider genomic regions of this large viral group. 相似文献