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Exogenous nitric oxide triggers Neospora caninum tachyzoite-to-bradyzoite stage conversion in murine epidermal keratinocyte cell cultures 总被引:1,自引:0,他引:1
Vonlaufen N Müller N Keller N Naguleswaran A Bohne W McAllister MM Björkman C Müller E Caldelari R Hemphill A 《International journal for parasitology》2002,32(10):1253-1265
Neospora caninum, like Toxoplasma gondii, undergoes stage conversion in chronically infected animals, and forms tissue cysts which contain the slowly proliferating bradyzoite stage. These tissue cysts are delineated by a cyst wall, protect the parasite from physiological and immunological reactions on part of the host, and bradyzoites remain viable within an infected host for many years. However, unlike T. gondii, N. caninum bradyzoites have been difficult to obtain using in vitro culture techniques, and current protocols, based on those developed for T. gondii, have been shown to be not very efficient in promoting tachyzoite-to-bradyzoite stage conversion. We report here an alternative in vitro culture method to obtain stage conversion of N. caninum from the proliferative to the cystic stage by using the Nc-Liverpool isolate, murine epidermal keratinocytes as host cells, and continuous treatment of infected cultures with 70 microM sodium nitroprusside for up to 8 days. This treatment significantly reduced parasite proliferation as assessed by Neospora-specific quantitative real-time PCR. The expression of bradyzoite markers was analysed by immunofluorescence following 4 and 8 days of in vitro culture using antibodies directed against bradyzoite antigen 1, the mAbCC2, and the lectin Dolichos biflorus agglutinin. Expression of the tachyzoite-specific immunodominant antigen NcSAG1 and the tachyzoite antigen NcMIC1 was also assessed. Transmission electron microscopy revealed that the majority of parasitophorous vacuoles were in the process of forming a distinct cyst wall through accumulation of granular material at the periphery of the vacuole, and parasites exhibited the typical features of bradyzoites. These findings demonstrate the usefulness of this culture technique as a promising way to study tachyzoite-to-bradyzoite stage conversion in N. caninum in vitro. 相似文献
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Stroehle A Schmid K Heinzer I Naguleswaran A Hemphill A 《The Journal of parasitology》2005,91(3):561-563
Toxoplasma gondii represents the most prominent infectious parasitic organism found in humans. While normally asymptomatic in healthy individuals, toxoplasmosis can cause abortion in patients during pregnancy, or can be fatal in immunosupressed individuals such as persons suffering from acquired immunodeficiency syndrom (AIDS). Toxoplasma gondii infection in humans is routinely assesssed by serological means. Here, we show that detection of anti-T. gondii IgG is also possible using a non-invasive methodology employing saliva. Sera and saliva of 201 healthy volunteers were investigated for the presence of anti-T. gondii-IgG antibodies by immunoblotting. The sera of 59 (29.4%) individuals showed IgG antibodies against T. gondii by ELISA, Vidas, and immunoblotting; 58 (98.3%) of these were also positive for anti-T. gondii IgG in the saliva immunoblot, with diagnostic relevant bands of Mr of 32-35 kDa and 40-45 kDa. The saliva immunoblot test exhibits a specificity of 100% and a sensitivity of 98.5%. Thus, saliva could be used as an alternative, non-invasive means for the detection of specific anti-T. gondii IgG in humans. 相似文献
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The ability to detect Odontoglossum ringspot virus (ORSV) released from the cut surfaces of leaf discs by ELISA was examined. Results indicate that ORSV from leaf discs can be detected but that multiple discs are necessary to obtain reliable detection of low virus concentrations, ELISA of Cymbidium protocorms, known from immunosorbent electron microscopy to be infected with ORSV, frequently produced A435 values which would be considered either negative or marginally positive on the basis of commonly accepted statistical limits (i, e. 2 x mean, or mean + 3 x standard deviation). The comparison of paired samples, one of which had been pretteated with ORSV antiserum, improved the sensitivity of the test from 10 ng ml ‘’ of virus to 2 ng ml’. 相似文献
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de Silva HA Pathmeswaran A Ranasinha CD Jayamanne S Samarakoon SB Hittharage A Kalupahana R Ratnatilaka GA Uluwatthage W Aronson JK Armitage JM Lalloo DG de Silva HJ 《PLoS medicine》2011,8(5):e1000435