Correlative study between immunodulatory action of propolis on cultured PBMNCs and it's schistosomicidal activity

Propolis, a beehive product widely used in folk medicine as an anti-nflammatory agent, have heen attracting researchers attention to scientifically elucidate us biological properties and therapeutic activities. This study aimed to spot light on the value of propolis as an immune-stimulant and to evaluate the influence on schistosome hematobium infection cure rate. To achieve this goal we estimated the effect of propolis on cultured peripheral blood mononuclear cells activation in-vitro by IL-2 and NO determination. We also evaluated the effect of in-vivo treatment with propolis on Schistosoma hematobium worm and bone marrow by parasitological and ultrastructural studies. Twenty S. haematobium infected golden hamsters were included in the study, subdivided into two groups each of 10 animals Group 1: Infected Control with 300 +/- 10 cercariae of S. haematobium by abdominal skin exposure. Group 2: Animals were treated with propolis three months post the infection. Our in-vitro results revealed that propolis induces a discreet elevation in IL-2 and NO release in PBMNCs cultures supernatant of S. hematobium infected hamsters. Mean level of IL-2 was 16.17 +/- 1.67 pg/ml in the presence of propolis and 3.31 +/- 0.76 in its absence with highly statistically significant difference [p < 0.001]. Regarding NO, Mean level of NO was 7. 76 +/- 1.30 U/ml in the presence of propolis and 2.6 +/- 0.42 in its absence with. highly statistically significant difference [p < 0.001]. Also, propolis caused observed activation and absence of apoptotic changes at the ultrastructural level of cultured PBMNCs revealed. In-viva results, revealed significant reductions in mature worm loads [either male or female], tissue egg loads [either intestinal or hepatic] 21.00 and 19.79% respectively and Percentage reductions of egg developmental stages was 68.07% with statistically significant difference compared with infected control group [P < 0.05]. Ultrastructural study of S. hematobium women revealed implantation and degeneration of the spines within vesiculated tegument and for the bone marrow it revealed evidence of lymphocyte and promonocyts activation in addition to remarkable increase in the number of the activated natural killer cell. Data suggest that propolis acts on host immunity by PBMNCs activation. This information would provide new insights in considering propolis to have a potential therapeutic benefit if used in conjunction with antischistosomal drug in treatment of schistosome infection

Parasitological, pathological, ultrastructural study of the effect of pentoxyifylline [PTX] anti-transforming growth factor beta [TGF beta] on S. inansoni infected mice

The main problem in schistosomal hepatic morbidity is fibrosis and extensive scarring induced by living eggs. The potential that pentoxifyllin [PTX] and anti-transforming growth factor- beta [anti-TGF beta] are anti-inflammatory and anti-fibrotic, urged us to study these effects parasitologically, pathologically and ultrastructurally in a murine model of schistosomiasis mansoni. Sixty albino mice were infected by subcutaneous injection of 80 +/- 20 S. mansoni cercariae/mouse. They were divided into two groups. Each group was subdivided and treated with PTX or anti- TGF beta for 3 weeks either at 4 weeks post infection [P.I] [1[St] group, acute phase] or at 12 weeks P.1. [2[nd] group, chronic phase]. Mice were sacrificed by the end of treatment [i.e. 7 and 15 weeks P.I. respectively]. A subgroup in each served as infected untreated controls. Parasitological assessment of worm burden, tissue egg load and oogram pattern was carried out. Measurement of granuloma diameter and ultrastructure of adult worms were also investigated. The study revealed decrease in the mean size of granuloma in mice treated with PTX and significant reduction in worm burden in all groups as compared to control group. Tissue egg load also significantly decreased, but no significant changes were observed in oogram pattern except an increase in number of dead ova in groups given anti- TGF beta 15 weeks following infection. Scanning electron microscopic examination of both male and female worms exposed to PTX treatment revealed tegumental changes in comparison to the control group. In conclusion effect of both PTX, anti. TGP beta regarding modulation of hepatic granulomas. Moreover it highlights thier role in produsing significant decrease in percent reduction of worm burden, egg load and increase dead ova

Ultrastructural study of peripheral blood mononuclear cells in chronic type B hepatitis

Many studies have suggested the infection of peripheral blood mononuclear cells [PBMCs] by hepatitis B virus [HBV] and its clinical implication in affecting the clinical status and persistence of the disease. However, there is still controversy concerning the mode of existence, replicative potential and cytopathological effect of the virus in these cells. In ultrastructural studies, three morphological forms of viral particles were detected in serum and hepatocytes of hepatitis B patients. However, none of these studies involved the PBMCs. In the present work, it was aimed to directly examine isolated PBMCs by transmission electronmicroscopy [TEM] to detect the various morphological forms of the virus and to verify any morphological alterations produced in the cells by the infection. PBMCs were isolated on ficoll-hypaque from heparinized blood of five patients with chronic hepatitis B [CHB] infection and subjected to conventional electronmicroscopy and immunoelectronmicroscopy [IEM]. Three healthy subjects were included as controls. Among PBMCs, many monocytes and lymphocytes showed either degenerative or apoptotic features. Dilatation of the Golgi apparatus [GA] and endoplasmic reticulum [ER] was also encountered. Some cells displayed intracytoplasmic particulate structures localized within vacuoles and appearing in various forms. One form resembled the complete virion but was larger [100 nm in diameter] and lacking the double-shell contour. Spherical HBsAg subviral forms [25-33nm in diameter] were also observed. These structures were verified as HBV particles by IEM. Our results have provided further evidence for the presence of the replicable complete virion-like particles in PBMCs. They seem to persist, replicate and induce injury to PBMCs. This may affect the clinical outcome and persistence of the infection

Mitochondrial cytochrome C and ultrastructural changes in hepatic steatosis induced by hepatitis C virus

Hepatic steatosis is an important hallmark of hepatitis C virus [HCV] infection. There is substantial body of evidence to implicate steatosis in the development of hepatic fibrosis. The underlying mechanisms of HCV-related steatosis however are not yet clarified. This study was performed to evaluate ultrastructural mitochondrial changes in patients with HCV-induced hepatic steatosis and correlate these findings with serum cytochrome c and apolipoproteins. Thirty-seven HCV-positive patients admitted to Theodore-Bilharz Research Institute were selected. They did not have other confounding prosteatogenic variables: diabetes, overweight, alcohol consumption and prosteatogenic drugs as amiodarone; corticosteroids. In addition, 10 apparently age- matched subjects were selected as a reference group. All subjects were initially subjected to full history, thorough clinical examination, liver function tests; lipid profile, HCV-IgG antibody and hepatitis-B surface antigen. All patients were subjected to ultrasound-guided liver biopsy. Biopsy specimen was processed for light and electron microscopic histopathological examination. According to histopathological findings, patients were divided into 4 groups according to the stage of fibrosis[I, II, III and IV]into :-Group 1 [n=7]; Group 2 [n=9]; Group:3 [n=9] and Group 4 [n=12]or cirrhotic group respectively. Measurement of apolipoproteins A, B and II and specific estimation of serum cytochrome-C was performed. Interpretation of the results revealed accumulated fat droplets by ultrastructure identification in the hepatocytes together with hypobetalipoproteinemia and hypotriglyceridemia. This was accompanied with mitochondrial ultrastructural alterations in all the studied groups ranging from complete dissolution to loss of outer mitochondria] membrane. In addition, it is noteworthy that ultrastructural changes of the rough endoplasmic reticulum [RER] detected in this study may be a contributing factor to abnormal fat metabolism in HCV. Concomittantly, serum cytochrome c was significantly lowered in all the studied groups as compared to the reference mean value. Depletion of mitochondria] cytochrme c might result in accumulation of reactive oxygen species and further accentuation of steatosis. There was significant correlation between serum cytochrome c, apolipoprotein B and serum triglycerides in the patients' group, ushering that it might have its role in HCV-induced lipid changes. In the cirrhotic group, ultrastructural elucidation of homogenous unlocalized intracytoplasmic fat and evident intracytoplasmic collagen fibrils was reported in this study. This was associated with significantly lowered serum ApoAl, ApoB and triglycerides as compared to all other groups. The afore-mentioned data solidifies the evidence that hepatic steatosis and fibrosis are important sequence of HCV infection and confirm the ability of hepatocyte to synthesize collagen. Moreover, the discerned intracytoplasmic unsaturated fat droplets might mirror the inability of the HCV-dysfunctioning hepatocyte to cope with the excessive dietary intake of fat and hence accentuation of steatosis results. In conclusion, ultrastructural. mitochondrial and RER changes seems to confirm their relation- to the associated intracytoplasmic fatty acids accumulation, the decreased serum cytochrome c and apolipoproteins Al and B Moreover it confirms the cytopathic effect of HCV