Hematological and parasitological effect of cyctein protease inhibitor on murine schistosomiasis

Cysteine protease enzyme [CP] is crucial for parasitic disease propagation, and inhibitors of such proteases are emerging with promising therapeutic uses in the treatment. This study was designed to evaluate the hematological and parasitological efficacy of one of the cysteine protease inhibitors [Sodium nitroprusside] that was administered alone as chemotherapy for murine schistosomiasis manosni in different schistosomal stages [shistosomula and mature worm]. Thirty mice were infected with 80 S. mansoni cercariae/mouse and were divided into 3 groups [10 mice each]. Group I: Infected untreated mice Group II and III. Mice were intraperitoneally treated with 100 mg/kg of cysteine protease inhibitor for 10 consecutive days three and six weeks post infection. Eight weeks post infection all animals were sacrificed and subjected for assessment of cysteine protease protein expression in liver tissue samples using immunoblotting technique [Western blotting], parasitological criteria and EM examination of buffy coat bone marrow and worms. Expression of cysteine protease protein was detected at the expected molecular weight [28 kDa] in 9 of the 10 [90%] infected untreated mice. After treatment protein expression returned negative in the treated groups. A highly significant reduction in worm burden was observed in all groups treated with inhibitor in comparison to infected control group [p<0.001]. However the greatest reduction in the worm burden was observed in treated group 6 weeks post infection in comparison to 3 weeks post infection [P<0.05]. Also, treatment could reduce egg count when given early in infection or significantly decreased egg burden when given late in infection. As regards EM examination sodium nitroprusside treated mice 3 and 6 weeks post the infection revealed degenerated tegument with completely implanted and degenerated spines. In addition 6 weeks post infection treated Schistosoma mansoni male worms showed vaculation and necrosis of spermatocytes. Buffy coat examination of Schistosoma mansoni infected untreated mice showed the inability of the eosinophil to be degranulated, while both treated groups revealed degranulation. In addition the group treated six weeks post infection showed hypodense eosinophils with large number of cytoplasmic vacuoles which represent an activated phenotype. Also in latter group activated lymphocytes were detected with marked dilatation of the endoplasmic reticulum. Bone marrow examination of Schistosoma mansoni - treated mice revealed degranulated eosinophils and eosinophilic myelocytes with activated phenotype in addition to degranulated platelets. Our findings indicate that the cysteine protease secreted by the different stages S. mansoni plays a crucial role in attenuating effector functions of eosinophils as it decreases the eosinophil's responses and inhibit its activation to the parasite resulting in diminished degranulation and reduced generation of superoxide. So, it is preferable to give CPI at any time post infection, with frequent observation of platelets function and different coagulation tests

Estimation of protease activity in Schistosoma mansoni life cycle stages and Biomphalaria Alexandria after cystein protease inhibitor

The efficacy of cysteine inhibitor on murine Schistosom mansoni was studied when the injection of inhibitor to infected mice for 10 successive days was either starting on day 1 [group 1] or on day 21 [group2] or on day 45 [group3] post infection. Eight weeks post infection animals were sacrificed and subjected for parasitological, histological and physiological assessments. Then levels of proteases activity were assayed in normal, infected and infected treated mice in serum, livers, intestine, as well as, in S. mansoni worms and ova collected from different groups under study. Also, B. alexandrina snails were exposed to LC5, LC10 and LC25 of each of bayluscide and A. arvensis then their hemolemph protease activity were assessed after 1 day, 3 days, 1 week, 2 weeks, 3 weeks and 4 weeks of exposure and compared with that assessed in hemolemph of control snails. Both the parasitological and physiological studies gave rise to the same conclusion denoting that treatment with cysteine protease inhibitor, phenyl vinyl sulfone, at 45 day post infection could reduce the worm burden, egg tissue load, granuloma diameter, ova hatchability, proteases activity in S. mansoni worms and ova. This treatment at the same time helps mice to attain approximately normal physiological state as expressed by their serum proteases activity. The findings representing toxicity and physiology studies gave rise to the same conclusion, emphasizing that the chemical molluscicides seriously affects snail physiology exerting irreversible alterations, while plant molluscicides has effect under which snails can acclimatize their physiology to survive

Ultrastructural and histopathological changes in Giardia lamblia-infected hamsters following administration of two natural compounds and/or metronidazol

The use of combined treatment of metronidazole together with arthemisia and rosemary for controlling intestinal Giardia lamblia infection has been studied. Forty laboratory-bred male hamsters were used in the current experimental study. Each hamster was infected, orally, by 10.000 Giardia lamblia cysts. Animals were divided into the following groups: [1] control infected untreated, [2] infected treated with metronidazole, [3] infected treated with arthemisia and rosemary, [4] infected receiving combined treatment of metronidazole plus arthemisia and rosemary. Each treatment was applied three weeks post-infection. Two weeks later stool analysis was performed and the number of cysts/gm stool was counted, followed by scarification of all animals. The effect of the different drug regimens on the vegetative forms [trophozoites] of the parasite was also studied. There was a highly significant cyst reduction in all treated groups compared to control animals. The highest percentage trophozoite reduction [98.7%] was found in group 4 receiving combined treatment of metronidazole plus arthemisia and rosemary, followed by group 2 [94.8%] and group 3 [62.2%]. By histology, healing of mucosal ulcerations, preserved villi and reduced chronic inflammatory infiltrate in the lamina propria were detected with combined therapy. Ultrastructurual examination of the small intestine in animals of control group showed destructed intestinal cell projections by Giardia lamblia cysts with degeneration of the intestinal submucosa. With combined treatment, complete repair of the intestinal cell projections as well as healing of the mucosa and the submucosa were noticed. Meanwhile, partial healing of destructed intestinal cell projections was observed in group 2 receiving metronidazole alone. It was concluded that the best results were obtained following combined treatment of metronidazole together with arthemisia and rosemary. This might be useful in endemic areas where people tend to develop drug resistance to the commonly used anti-Giardia lamblia preparations

Parasitological, hematological and ultrastructural study of the effect of Cox-2 inhibitor, pyocyanin pigment and praziquantel, on S. mansoni infected mice

The effect of cyclooxygenase-2 [COX-2] inhibitor, such [as meloxicam, and pyocyanin pigment of Pseudomonas aeruginosa] with and without praziquantel [PZQ] on worms, ova count, bone marrow and blood cells in 7 groups of Schistosoma mansoni infected mice was studied. The results revealed significant decrease of worm burden and ova count in all treated groups as compared to the infected untreated group, while those with combined treatment of PZQ and meloxicam or pyocyanin showed complete eradication of the worm with the highest reduction in the tissue egg load. EM showed extensive swelling and vesiculation of the tegument, completely implanted spines that overlie degenerated muscle layer were obvious in groups treated with either meloxicam or pyocyanin. Hematological study revealed significant increase [P < 0.05] of total leucocytic count of PZQ treated group while that treated with either meloxicam or pyocyanin showed significant decrease [P < 0.05], but in combination of PZQ with meloxicam or pyocyanin no significant difference as compared to the infected untreated group. The neutrophil was the main cell affected in groups treated with neither meloxicam nor pyocyanin alone with significant decrease [P < 0.05], but with significant increase [P < 0.05] in combination with PZQ as compared to the infected untreated group. Those treated with PZQ plus meloxicam showed significant increase as compared to that plus pyocyanin. Eosinophil count showed significant decrease [P < 0.05] in all treated groups as compared to the infected untreated group. Inverse correlation between serum level of sFas and peripheral neutrophil count was detected. Ultrastructural study of the bone marrow explained the results as groups treated with meloxicam revealed dissociation between nuclear and cytoplasmic development in the neutophils with cytoplasm maintaining primitive appearance despite maturation of the nucleus that is manifested by the persistent production of immature granules and the still orientation of Golgi cternae and the centriole around the nucleus. Groups treated with pyocyanin pigment revealed many abnormalities in neutophils as hypogranularity or early apoptotic morphology changes as intense pen- nuclear chromatin aggregation or nucleus fragmentation.In peripheral blood apoptotic morphology changes was detected in both groups treated with meloxicam or pyocyanin while most of cells of mice treated with PZQ were in an active state. Consequently, it is preferable to give meloxicam with PZQ for a short period of time [less side-effect] to eradicate S. mansoni worm completely but with continuous observation of the peripheral neutrophil count and function