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Experimental parasitology [journal]
- In vitro evaluation of 4-phenyl-5-(4́-X-phenyl)-1,3,4-thiadiazolium-2-phenylaminide chlorides and 3[N-4́-X-phenyl]-1,2,3-oxadiazolium-5-olate derivatives on nitric oxide synthase and arginase activities of Leishmania amazonensis. [JOURNAL ARTICLE]
- Exp Parasitol 2013 May 18.
AbstractLeishmaniasis is a spectrum of infectious diseases caused by Leishmaniaprotozoan parasites. The purpose of this study was to perform, in vitro, a comparativeanalysis of the activity amastigotes. Results showed excellent efficacy of all compounds against axenic amastigotes, compared to pentamidine isethionate, the reference drug used. The cytotoxic effect of these mesoionic compounds of six mesoionic compounds (three 1,3,4-thiadiazolium-2-aminide and three 1,2, 3oxadiazolium-5-olate class compounds) against Leishmania amazonensis promastigotes and axenic expressed, was evaluated in mouse peritoneal macrophages using MTT assay, low toxicity (∼10%) for these mammalian cells being observed. In an attempt to define a potential drug target, the activities of nitric oxide synthase (NOS) and arginase on the parasites treated with the mesoionic derivatives were evaluated. NOS was purified from a cell-free extract of infective promastigotes and axenic amastigotes and all derivatives tested were able to inhibit the enzyme as monitored by the decrease of NADPH consumption. Arginase activity from both stages of the parasite was measured using urea production and none of the compounds inhibited the enzyme activity of axenic amastigotes. On the other hand, when tested with promastigotes, those compounds without MI-H and SID-H substituents inhibited arginase activity.
- Schistosoma japonicum scFv-IL18 fusion DNA ameliorates hepatic fibrosis in schistosomiasis-infected mice via improving local concentration of IL-18 in liver. [JOURNAL ARTICLE]
- Exp Parasitol 2013 May 14.
The pathogenesis of chronic schistosomiasis is caused by irritation of the schistosome eggs trapped in liver that induce delayed hypersensitive reactions from the surrounding tissues, leading to the formation of inflammatory granuloma and subsequent fibrosis. A Schistosoma japonicum (S. japonicum) single-chain fragment variable (SjscFv) which specifically binds to the S. japonicum soluble immature egg antigen (SIEA) can be used as a target to deliver specific cytokine towards the site of hepatic fibrosis. To test this hypothesis, a novel recombinant plasmid, pVAX1/SjscFv-IL18, was constructed by fusing SjscFv to IL-18 gene with a 45bp glycine-rich linker. Furthermore, experiments on mice showed that pVAX1/SjscFv-IL18 could effectively express IL-18 in the liver and in serum. Hepatic contents of IL-2 and IFN-γ (Th1-type) in S. japonicum-infected mice vaccinated with pVAX1/SjscFv-IL18 increased significantly but those of their IL-4 and IL-10 (Th2-type) decreased as compared to the analyzed results of 4 cytokines in the liver cells of control mice vccinated with pVAX1/IL18. Consistent with the levels of Th1 and Th2 cytokines, mice vaccinated with pVAX1/SjscFv-IL18 developed much less hepatic fibrosis 20weeks after infection, which was evaluated by average volumn of granuloma and collagen contents. These data suggested that the linkage of IL-18 to the target-specific SjscFv molecule appears to be a potentially promising trial route of therapy, the hepatic fibrosis in S. japonicum-infected mice may be ameliorated through effective expression of IL18 in liver.
- Disruption of the blood-brain barrier in pigs naturally infected with Taenia solium, untreated and after anthelmintic treatment. [JOURNAL ARTICLE]
- Exp Parasitol 2013 May 15.
Neurocysticercosis is a widely prevalent disease in the tropics that causes seizures and a variety of neurological symptoms in most of the world. Experimental models are limited and do not allow assessment of the degree of inflammation around brain cysts. The vital dye Evans Blue (EB) was injected to 11 pigs naturally infected with Taenia solium cysts to visually identify the extent of disruption of the blood brain barrier. A total of 369 cysts were recovered from the 11 brains and classified according to the staining of their capsules as blue or unstained. The proportion of cysts with blue capsules was significantly higher in brains from pigs that had received anthelmintic treatment 48 and 120h before the EB infusion, indicating a greater compromise of the blood brain barrier due to treatment. The model could be useful for understanding the pathology of treatment-induced inflammation in neurocysticercosis.
- Role of TNF in sickness behavior and allodynia during the acute phase of Chagas' disease. [JOURNAL ARTICLE]
- Exp Parasitol 2013 May 14.
Chagas disease, caused by the intracellular protozoan Trypanosoma cruzi, is associated with inflammation, discomfort and pain during the acute phase. The influence of TNF-α (tumor necrosis factor) in this disease outcome is controversial. In this way, the aim of this work was to determine the role of the TNF-α blocker etanercept in the pain, discomfort, and survival during the Chagas' acute phase of mice experimentally infected with a wild virulent strain of T. cruzi. The infection with this wild strain was responsible for a severe visceral inflammation and said parasite showed a tropism in peritoneal fluid cells. Etanercept was able to restore spontaneous vertical and horizontal activities during the second week after infection and to abolish mechanical allodynia during the first week after infection. Finally, etanercept delayed the mortality without any effect on the parasitemia rates. This is the first report that correlates sickness behavior and allodynia with TNF-α and suggests that this cytokine may play an important role in the physiopathology of the acute phase.
- Pomegranate (Punica granatum) peel is effective in a murine model of experimental Cryptosporidium parvum. ultrastructural studies of the ileum. [JOURNAL ARTICLE]
- Exp Parasitol 2013 May 15.
The current treatments for cryptosporidiosis are ineffective, and there is an urgent need to search for more effective and safer alternatives. One such alternative may be treatments derived from natural resources. The pomegranate peel has been used effectively in traditional medicine to cure diarrhea and dysentery. The purpose of this study was to examine the effectiveness of a Punica granatum (pomegranate) peel suspension as a treatment for Cryptosporidium parvum infection. In this study, the effects of this treatment on the ultrastructure of both the intestinal epithelial layer of infected nursling mice and the parasite were observed with a transmission electron microscope. The histological study focused on the examination of the microvilli, columnar epithelium, goblet cells, lamina propria, and crypts of Lieberkuhn. Examination of the ileums of infected mice that received the pomegranate peel suspension demonstrated that the general structure of the ileal tissue of these mice was similar to that of the control group. In the infected mice treated with the suspension, but not the infected/untreated mice, there was an improvement in all ultrastructure aspects at 28 days post-inoculation. The study of the ultrastructure of the parasite (C. parvum) in mice treated with the suspension showed that there was decomposition in the parasite to the extent that in some cases we were unable to identify the stage of the parasite due to the severe degeneration. Significant decomposition of the nutrition organ was also observed. Additionally, microgamonte and macrogamonte were not observed in the suspension-treated group, explaining the disappearance of the sexual phases of the parasite in the lumens of this group. In all, this examination demonstrated the restoration of the normal structures of villi and the disappearance of acute symptoms in the suspension-treated mice and showed that the suspension directly affected the parasite at various stages of its development and led to its decomposition and death.
- Impact of Schistosoma mansoni and Echinococcus granulosus experimental coinfection on interleukin 10 and Interferon gamma cytokines profile. [JOURNAL ARTICLE]
- Exp Parasitol 2013 May 15.
The interactions between various aspects of the immune responses mediated by concomitant parasite infections may influence the resultant cytokines profiles. We tested this hypothesis by developing two Schistosoma mansoni and Echinococcus granulosus coinfection murine models. Our aim was to explore the effect of echinoccocis on the immune responses induced by schistosomiasis, either when the two infections were induced synchronously or when echinococcosis was induced during egg deposition period of S.mansoni infection. The proliferation of antigens specific stimulated splenocytes, taken from studied groups, was determined. Then, IFN-γ, and IL-10 production from stimulated cells were measured. Significant elevation of IFN-γ, four weeks after synchronous coinfection, was occurred compared to S.mansoni infected group, associated with modest elevation of IL10 level. On the other hand, echinococcosis coinfection during egg deposition period of schistosomiasis resulted in significant marked reduction in IL10 level in comparison to S.mansoni infected mice. These results suggested that echinococcosis coinfection, during the switching from Th1 to Th2 cytokine stage of murine schistosomiasis, can alter the ability of S.mansoni infection to skew the cytokines response towards Th2 profile. It is clear that the timing and sequence of concomitant infections are of vital importance for the outcome of the immune response γ
- Entamoeba histolytica antigenic protein detected in pus aspirates from patients with amoebic liver abscess. [JOURNAL ARTICLE]
- Exp Parasitol 2013 May 13.
Entamoeba histolytica is a causative agent of amoebic liver abscess (ALA) and is endemic in many underdeveloped countries. We investigated antigenic E. histolytica proteins in liver abscess aspirates using proteomics approach. Pus samples were first tested by real-time PCR to confirm the presence of E. histolytica DNA and the corresponding serum samples tested for E. histolytica-specific IgG by a commercial ELISA. Proteins were extracted from pus samples which were pooled from three and one patients with ALA and PLA (pyogenic liver abscess) respectively, followed by analysis using isoelectric focussing, SDS-PAGE and Western blot. Unpurified pooled serum samples from infected hamsters and pooled human amoebic-specific IgG were used as primary antibodies. The antigenic protein band was excised from the gel, digested and analysed by MALDI-TOF/TOF and LC-MS/MS. The results using both primary antibodies showed an antigenic protein band of ∼14kDa. Based on the mass spectrum analysis, putative tyrosine kinase is the most probable identification of the antigenic band.
- Evidence of apoptosis in Raillietina echinobothrida induced by methanolic extracts of three traditional medicinal plants of Northeast India. [JOURNAL ARTICLE]
- Exp Parasitol 2013 May 13.
The therapeutic benefits of medicinal plants in terms of anthelmintic properties are known since time immemorial in India, particularly among natives of the Northeast India. However, only sporadic and scarce reports on scientific validation of these plants are available. The present study was conducted on the cestode Raillietina echinobothrida, to establish whether the anthelmintic activity of Potentilla fulgens, Alpinia nigra and Millettia pachycarpa was mediated by apoptosis or not. Light microscopic observation following MTT assay revealed the highest percentage of inhibition of viability among the worms by methanol extract of M. pachycarpa (89.33%), followed by A. nigra (65%) and P. fulgens (37%). Ultrastructural observations revealed swelling of mitochondria, disruption of mitochondrial membrane, vacuolization of mitochondria, appearance of apoptotic bodies in the cytoplasm, disintegration of nuclear membrane and nucleolus were very common throughout the tegument. DAPI stained specimens showed typical morphology of apoptosis, like nuclear condensation and fragmentation in the extracts treated parasites. A decrease in mitochondrial membrane potential was also recorded in the treated groups. Confirmatory TUNEL assay and DNA fragmentation assay of the extracts treated parasites also confirmed the apoptotic nature of cell death and is concluded to be responsible for paralysis and death of the parasite.
- Neospora caninum: Infection induces high lysosomal activity. [JOURNAL ARTICLE]
- Exp Parasitol 2013 May 3.
Neospora caninum is a protozoan that causes abortion in cattle and neuromuscular lesions in dogs, making it an important target of veterinary medicine. Lysosomes are cellular organelles responsible for important biological functions as cellular defense mechanisms. The aim of this work was to evaluate the lysosomal stability of rat gliocytes infected in vitro with N. caninum. Rat glial cultures were infected at a ratio of 1:1 (cell/parasite). The enzymatic activity of acid phosphatase (orthophosphoric-monoester phosphohydrolase, EC 18.104.22.168) was assayed in the medium of control and infected cell cultures. The activity observed at 24h of incubation was 0.4±0.08mU/mg/min for control cells and 1.3±0.5mU/mg/min for infected cells. After 72h, control and infected cells exhibited activities of 1.3±0.5 and 4.1±0.9mU/mg/min, respectively. These results suggested that lysosomal compartment plays an important role in the mechanisms of cellular infection by N. caninum.
- Ability of TEP1 in intestinal flora to modulate natural resistance of Anopheles dirus. [JOURNAL ARTICLE]
- Exp Parasitol 2013 May 3.
Blocking transmission of malaria is a reliable way to control and eliminate infection. However, in-depth knowledge of the interaction between Plasmodium and mosquito is needed. Studies suggest that innate immunity is the main mechanism inhibiting development of malaria parasites in the mosquito. Recent studies have found that use of antibiotics that inhibit the mosquito gut flora can reduce the immune response of Anopheles gambiae, thereby contributing to the development of malaria parasites. In our study, we used the non susceptible model of Anopheles dirus-Plasmodium yoelii to explore the effect of Anopheles intestinal flora on the natural resistance of A. dirus to P. yoelii. We found that in mosquitoes infected with Plasmodium, the intestinal flora can regulate expression of thioester-containing protein (TEP1) via an RNAi gene-silencing approach. Our results suggest that in the absence of TEP1, the natural microbiota cannot suppress the development of P. yoelii in A. dirus. This suggests that AdTEP1 plays an important role in the resistance of A. dirus to P. yoelii. The intestinal flora may modulate the development of P. yoelii in A. dirus by regulating TEP1 expression.