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Pasteurella multocida A:3 is a major cause of bovine pneumonia. A major antigenic heat-modifiable 28kDa outer membrane protein (Omp28) was previously identified. The purpose of this study was to purify and characterize Omp28 immunologically and structurally. Omp28 was extracted from N-lauroylsarcosine-insoluble protein preparations by a combination of detergent fractionation with Zwittergent 3-14 and chromatography. Partial N-terminal amino acid sequence confirmed Omp28 as a member of the OmpA-porin family. However, porin activity could not be demonstrated in a lipid-bilayer assay. Heat modifiability of purified Omp28 was demonstrated, and Omp28 was found in outer membrane fraction of P. multocida. Surface exposure of Omp28 was demonstrated by partial protease digestion of intact bacteria, by binding of anti-Omp28 polyclonal ascites fluid to the bacterial surface, and by partial inhibition of anti-outer membrane antiserum binding by previous incubation of the bacteria with anti-Omp28 serum. CD-1 mice vaccinated with purified Omp28 developed a significant antibody titer (P<0.05) compared to the control treatment group but were not protected from a homologous intraperitoneal bacterial challenge. By contrast, treatment groups vaccinated with P. multocida outer membrane, formalin-killed P. multocida or a commercial vaccine were significantly protected from challenge. In vitro complement-mediated killing of P. multocida was observed in post-vaccination sera of outer membrane, formalin-killed P. multocida, and commercial vaccine-treatment groups, but not with sera from the Omp28-treatment group. In conclusion, although Omp28 is surface exposed and antigenic, it may not be a desirable immunogen for stimulating immunity to P. multocida. 相似文献
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Sugars in the form of monosaccharides, oligosaccharides, polysaccharides and glycoconjugates (glycoproteins, glycolipids) are vital components of infecting microbes and host cells, and are involved in cell signalling associated with modulation of inflammation in all integumental structures. Indeed, sugars are the molecules most commonly involved in cell recognition and communication. In skin, they are essential to epidermal development and homeostasis. They play important roles in microbial adherence, colonization and biofilm formation, and in virulence. Two groups of pathogen recognition receptors, C-type lectins (CTL) and their receptors (CTLR), and the Toll-like receptors enable the host to recognize pathogen-associated molecular patterns (PAMPs), which are mainly glycolipids. The CTLs can recognize a wide variety of bacteria, fungi and parasites and are important in phagocytosis and endocytosis. TLRs are expressed on the surfaces of a variety of cells, including keratinocytes, dendritic cells, monocytes and macrophages; they play a major role in innate immunity. Interaction of TLRs with PAMPs initiates a cascade of events leading to production of reactive oxygen intermediates, cytokines and chemokines, and promotes inflammation. Exogenous sugars can block carbohydrate receptors and competitively displace bacteria from attachment to cells, including keratinocytes. Thus sugars may provide valuable adjunctive anti-inflammatory and/or antimicrobial treatment. A promising approach is the use of a panel of carbohydrate derivatives with anti-adhesive efficacy against bacteria frequently involved in diseases affecting skin and other epithelia. More complete characterization of sugar receptors and their ligands will provide further keys to use of carbohydrates in immunomodulation and infection control in skin. 相似文献
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V Furlanetto G Zagotto R Pasquale S Moro B Gatto 《Journal of agricultural and food chemistry》2012,60(36):9162-9170
Ellagic acid (EA), a natural polyphenol abundant in fruits and common in our diet, is under intense investigation for its chemopreventive activity resulting from multiple effects. EA inhibits topoisomerase II, but the effects on the human enzyme of urolithins, its monolactone metabolites, are not known. Therefore, the action of several synthetic urolithins toward topoisomerases II was evaluated, showing that polyhydroxylated urolithins, EA, and EA-related compounds are potent inhibitors of the α and β isoforms of human topoisomerase II at submicromolar concentrations. Competition tests demonstrate a dose-dependent relationship between ATP and the inhibition of the enzyme. Docking experiments show that the active compounds bind the ATP pocket of the human enzyme, thus supporting the hypothesis that EA and polyhydroxylated urolithins act as ATP-competitive inhibitors of human topoisomerase II. 相似文献
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Alexandra C. Buckley Shollie M. Falkenberg Mitchell V. Palmer Paulo H. Arruda Drew R. Magstadt Kent J. Schwartz Igor R. Gatto John D. Neill Bailey L. Arruda 《Journal of veterinary diagnostic investigation》2021,33(5):952
Atypical porcine pestivirus (APPV) is a cause of congenital tremors (CTs) in piglets and has been found in swine populations around the globe. Although systemic distribution of the virus has been reported, there is limited information regarding viral localization at the cellular level and distribution at the tissue level. We collected multiple tissues from 2-d-old piglets (n = 36) born to sows inoculated at 45 or 62 d of gestation with APPV via 3 simultaneous routes: intravenous, intranasal, and directly in amniotic vesicles. In addition, 2 boars from APPV-inoculated sows with CT were raised and euthanized when 11 mo old. In situ hybridization performed on tissue samples from piglets demonstrated a broad and systemic distribution of viral RNA including endothelial cells, fibroblasts, and smooth muscle. Labeling in tissues was more pronounced in piglet tissues compared to boars, with the notable exception of diffuse labeling of the cerebellum in boars. Presence of APPV in boar tissues well after resolution of clinical signs suggests persistence of APPV similar to other pestiviruses. 相似文献
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Activity of extracts from wild edible herbs against postharvest fungal diseases of fruit and vegetables 总被引：1，自引：0，他引：1
Maria Antonia Gatto Antonio Ippolito Vito Linsalata Nicholas A. Cascarano Franco Nigro Sebastiano Vanadia Donato Di Venere 《Postharvest Biology and Technology》2011,61(1):72-82
The use of plant extracts could be a useful alternative to synthetic fungicides in the management of rot fungi during postharvest handling of fruit and vegetables. The aim of this study was to assess the in vitro and in vivo activity of extracts obtained from nine wild edible herbaceous species (Borago officinalis, Orobanche crenata, Plantago coronopus, P. lanceolata, Sanguisorba minor, Silene vulgaris, Sonchus asper, Sonchus oleraceus, and Taraxacum officinale) against some important postharvest pathogens, i.e. Botrytis cinerea, Monilinia laxa, Penicillium digitatum, P. expansum, P. italicum, Aspergillus carbonarius, and A. niger. Phenolic composition of all extracts was evaluated by HPLC. Several derivatives of caffeic acid, of the flavones apigenin and luteolin, and of the flavonols kaempferol and quercetin, were identified. Extracts from S. minor and O. crenata showed the highest efficacy in all the trials. In particular, S. minor completely inhibited in vitro the conidial germination of M. laxa, P. digitatum, P. italicum, and A. niger and strongly reduced those of B. cinerea; O. crenata extract showed a lower but still significant reduction of conidial germination on all the tested fungi. Moreover, the extracts from both species were effective in reducing the germ tube elongation also when a slight inhibition of conidial germination was observed. In many cases, a dose effect was observed, with an increase of antifungal activity as the phenolic concentration increased. In trials performed on wounded fruit, S. minor extract completely inhibited brown rot on apricots and nectarines; O. crenata extract strongly reduced grey mould, brown rot, and green mould on table grapes, apricots and nectarines, and oranges, respectively. The inhibition efficacy of extracts was ascribed to the presence of some caffeic acid derivatives and/or flavonoids. HPLC phenolic analyses provided useful information to identify the possible active compounds. 相似文献
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Canine coronavirus induces apoptosis in cultured cells 总被引：1，自引：0，他引：1
Ruggieri A Di Trani L Gatto I Franco M Vignolo E Bedini B Elia G Buonavoglia C 《Veterinary microbiology》2007,121(1-2):64-72
Canine coronavirus (CCoV) is widespread in dogs in several countries and causes mild enteric illness evolving to severe enteritis in young pups. In in vitro cultures canine coronaviruses generally induce extensive cell death, however nature of the events leading to cell death remains largely unknown. We analysed the induction of cytopathic effect by CCoV in a canine fibrosarcoma cell line (A-72) in order to characterize the apoptotic effect in homologous cell system. Following CCoV infection A-72 cell line, which is permissive to CCoV, showed reduced growth rate, as detected by MTT assay, a standard colorimetric assay for measuring cellular proliferation, and underwent to apoptotic death. Starting from 24h after CCoV infection, cells morphology appeared dramatically changed, with cells rounding and detachment from culture surface. Morphologic and biochemical features of apoptosis, such as blebbing of the plasma membrane, translocation of phosphatidilserine to cell surface and annexin V positive staining, nuclear fragmentation, apoptotic bodies formation and DNA laddering, were detected in CCoV-infected cells. Propidium iodide staining of infected culture indicated the appearance of hypodiploid DNA peak corresponding to apoptotic cell population. Commonly to other animal coronavirus infection caspase-3 is likely to contribute to the execution phase of apoptosis induced by CCoV in A-72 cells since we found activation of enzymatic activity as well as procaspase-3 activating cleavage. Apoptotic death of infected cells is detrimental as it causes cell and tissue destruction as well as inflammatory responses. Therefore in the case of CCoV associated gastroenteritis, apoptosis of epithelial mucosa cells may be responsible for pathology induced by CCoV infection. 相似文献
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de Mora F de la Fuente C Jasmin P Gatto H Marco A Ferrer L Fondati A Fondevila D Torres R 《Veterinary immunology and immunopathology》2007,115(3-4):223-229
Canine atopic dermatitis (AD) is a pruritic skin condition that shares many clinical and pathophysiological features with its human counterpart. A major therapeutic challenge of AD is the control of the skin inflammatory process. A detailed knowledge of the pro-inflammatory molecules involved in cell recruitment in AD would allow for a better control of the disease. We thus have studied the protein expression of P-selectin, ICAM-1 and TNF-alpha in the lesional and non-lesional skin of atopic dogs that had been treated for bacterial infections. Despite a low-to-mild inflammatory process, P-selectin protein was clearly upregulated in the lesional skin areas when compared with non-lesional skin (four-fold average increase). This P-selectin upregulation was accompanied by signs of functional changes such as increased cell margination, and membrane-associated protein expression. Although the expression of ICAM-1 and TNF-alpha was not enhanced in the lesional versus the non-lesional skin, there was a trend towards a correlated upregulation of both molecules. Further studies will help elucidate the significance of the substantial overexpression of P-selectin in canine AD, in particular in a scenario where bacterial antigens are not contributing as pro-inflammatory stimuli. 相似文献
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