الفهرس 
TitleOnly 14 pages are availabe for public view
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Abstract
Schistosomiasis is remaining an important public health problem in many developing countries including Egypt. Indeed, the disease exerts heavy medical, economic, and psychosocial impacts on the populations in endemic regions. Schistosomiasis elimination believed to be continued in the near future; therefore, awareness of the different aspects of the disease is still mandatory in endemic countries.
Schistosomiasis has plagued the Egyptian population since the antiquity. Continues sustained effort reduced morbidity and transmission of schistosomiasis. This effort focuses on periodic, targeted treatment with PZQ through the large-scale treatment of the affected populations via mass chemotherapy. As a result of this large-scale use of PZQ, an evidence of developing resistance to the drug has been identified. This evidence has found a definite confirmation by recent reports carried out in Egypt. Moreover, the drug has the disadvantage of inability to prevent reinfection as well as the lack of efficacy against juvenile schistosomes. One way to offset the emergence of drug resistance is to use alternative treatment regimens.
In the light of the above, the current work aimed to examine the differentially expressed proteins in S. mansoni infected mice before and after treatment with ivermectin and praziquantal based on proteomic analysis.
In the present work, 60 male Swiss Albino Mus musculus mice were recruited for this study from Theodor Bilharz Research Institute, Giza, Egypt. All processes were performed in agreement with the Institutional Animal Care and Use Committee, Alexandria University, Egypt. Mice were equally divided into three main gps (20 mice per each gp), each main group was further sub divided into two sub-gps (a&b) (10 mice per each sub-gp) according to numbers of cercariae used for infection, type of treatments and time of scarification.
Summary, conclusion & Recommendations
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Parasitological studies on studied sub-gp (b) in each mice gps were done to assess the efficacy of both PZQ and IV drugs by recording worm burdens, oogram patterns in intestine and ova counts per gram tissues (livers and intestines of mice).
Histopathological studies were performed on dissected livers. Samples were stained with H&E stain to evaluate pathological changes and other smears samples were stained by Masson’s trichrome stain for further granuloma classification.
Schistosoma adult worms recovered from the infected untreated sub-gp (a) I and treated mice sub-gp (a) of gps II and III were subjected to SEM examination to demonstrate the effect of studied drugs (PZQ and IV) on morphology of tegumental surfaces of the adult worms.
Proteomics approaches encompass the most efficient and powerful tools for identification of protein complexes. Proteomic analysis started by proteins extraction from recovered adult worms harvested of the three studied sub-gp (a) followed by SDS-PAGE for further protein separation. Proteins band were individually excised from gel and digested with trypsin for their identification and quantification using mass spectrometry analysis. Finally mass spectrometry data analysis performed by conducting online search against Schistosoma spp. specific proteins databases UniProt (http://www.uniprot.org), and Scientific Data Sharing Information Bio information (http://lifecenter.sgst.cn/ protein). In addition to molecular function, biological process was assigned for the proteins identified according to information obtained from the Gene Ontology (GO) database.
The data were entered, verified and analyzed using SPSS software using appropriate statistical tests. Differences and associations were considered statistically significant at P <0.05.
After conducting the laboratory methods and appropriate statistical processing of the data obtained, the results revealed throughout this study can be summarized as follows: regarding worm burdens, copula and males showed a statistically significant reduction in the mean worm loads recovered from sub-gps (b) of gp II (PZQ) and gp III (IV) compared with sub-gp (b) infected control. But mean female worm counts showed no statistical significance with either PZQ or IV compared with control. Total worm loads showed a reduction percentages by 88%, 37.5% in mice sub-gp (b) treated by PZQ & IV respectively in comparison to its control worm loads.
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Oogram pattern showed an absence of immature ova in PZQ treated mice sub-gp (b) also a highly reduced mean percentage in IV treated mice sub-gp (b) (28.20± 4.87) in comparison to infected untreated control sub-gp (b) (51± 4.52). As regard mature ova, a great mean counts reduction recorded in both treated sub-gps (b) per gp II and III compared to control sub-gp (b). While dead ova showed a remarkable increase in PZQ treated mice (84.50±3.92) followed by IV treated mice (27± 3.76) in comparison to infected untreated control (5.90±1.29). All results showed statistical significance. Ova counts revealed a statistical significant reduction in the hepatic and intestinal tissues in both treated sub-gps (b) in comparison with control sub-gp (b) but reduction of ova counts more in hepatic tissue of mice sub-gps treated with PZQ than sub-gp (b) treated with IV.
Histopathological examinations confirmed the existence of granuloma around the presented ova and described the effect of both treatments on granuloma and hepatocytes surrounding it. PZQ revealed reduced sized granuloma and lessened inflammatory reactions in addition to hepatocytes enhancement. In a promising way IV showed a reduction in granuloma counts and mild inflammation. Fibro-cellular granuloma predominates in all treated gps [PZQ and IV treated sub-gp (b)].
As for SEM examination on adult worms collected, a grievous tegumental destruction, peeling, erosion, ulceration and suckers damage recorded in PZQ treated adult worms harvested and examined. Whereas tegumental damaging effects on IV treated adult worms were to a lesser extent than PZQ treated worms.
Identification of proteins is very important for understanding how schistosomes regulate host immune systems to establish chronic infections and also elucidate other aspects of parasite-host interaction. Proteomic analysis revealed notable protein profile changes and differential protein intensity between studied sub-gps (a). Total lab analysis software document 19 protein bands, 12 commonly shared proteins between all studied sub-gps (a) and seven differential protein bands.
Noticeable disappearance recorded by SDS electrophoresis of two protein bands in both treated sub-gps (a), disappearance of another one protein band in PZQ treated sub-gp (a) rather than IV and control sub-gps (a) in addition to disappearance of three proteins in IV sub-gp (a) rather than PZQ and control sub-gp (a) . Furthermore, one protein band were detected in PZQ sub-gp (a) and absent from either IV or control sub-gps (a).
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Mass spectrometry identified the seven differential protein bands which expressed differentially between all studied sub-gps adult worms. According to molecular weights of separated protein bands, molecular and biological function administrated from NCBInr database showed the presence of glycolytic proteins, structural proteins and cytosol stress response chaperones.
These data provide new insights into the possible use of IV as a substitute for PZQ drug. Also spotted the light on the critical role of proteomic analysis in providing and emphasizing the complete depiction of structural and functional information of cell as well as the response mechanism of cell against various types of stress and drugs.