Assessment of spiramycin-loaded chitosan nanoparticles treatment on acute and chronic toxoplasmosis in mice.

Toxoplasmosis is a zoonotic parasitic disease with worldwide distribution. Chitosan is a natural polymer which is commonly used in the production of nanomedicines. It is known to enable higher drug permeation, being biocompatible and has very low toxicity, besides its antimicrobial effects. Our study aimed to assess the effect of spiramycin-loaded chitosan nanoparticles (SLCNs) in treatment of acute and chronic toxoplasmosis in mice. 200 male Swiss albino mice were included in our study, divided to two main groups; Toxoplasma gondii RH strain infected group and ME49 strain infected group, each main group was subdivided into four subgroups; subgroup I: infected control, subgroup II: infected and received chitosan nanoparticles (CS NPs); 20 µg of CS NPs in 100 µl of PBS/mouse/dose, subgroup III: infected and treated with spiramycin (Rovamycin); 100 mg/kg/day, subgroup IV: infected and treated with 100 mg/kg/day spiramycin-loaded chitosan nanoparticles. Effect of treatment was assessed parasitologically and histopathologically. It was noticed that SLCNs significantly decreased the mortality rate of infected mice with both strains compared to high mortality rate of mice in the infected control subgroups. Moreover, there was a significant decrease in the number of organisms of SLCNs treated subgroup as compared to the other subgroups. Histopathological studies showed a marked improvement of the pathological pictures of brain, liver, spleen and eye in the subgroup received SLCNs as opposed to other groups. In conclusion, the present study revealed that loading of spiramycin on chitosan nanoparticles increased its antiparasitic effect on acute and chronic T. gondii infection.

Protein Z and Endothelin-1 genetic polymorphisms in pediatric Egyptian sickle cell disease patients.

BACKGROUND: Sickle cell disease (SCD) is a monogenic disease associated with multisystem morbidity. Vasculopathy caused by delicate imbalance between coagulation and endothelial systems plays a pivotal role in disease course. As Protein Z and Endothelin-1 genetic polymorphisms may increase the thrombotic risk, the aim of the current work was to verify the possible impact of Protein Z (PROZ G79A) and Endothelin-1 (EDN1 G5665T) polymorphisms on the clinic-laboratory features of the SCD in a cohort of Egyptian pediatric patients. METHODS: Genotyping of Protein Z G79A and Endothelin-1 G5665T was carried out by polymerase chain reaction-restricted fragment length polymorphism (PCR-RFLP) assay for 100 SCD patients and 100 controls. RESULTS: Protein -Z G79A polymorphism was not associated with vascular complications in the studied SCD patients. Endothelin-1 G5665T polymorphism was associated with pulmonary dysfunction (pulmonary artery hypertension and acute chest syndrome) and severe vaso-occlusive crises (VOC). CONCLUSION: Endothelin-1 G5665T polymorphism could be considered as a molecular predictor for pulmonary dysfunction and severe VOC in SCD. Further researches with larger cohorts are recommended to understand the pathophysiology of SCD and to explain the inter-patients' variability of disease severity.

CHARACTERIZATION OF THE HEMOCYTES OF SUSCEPTIBLE AND RESISTANT BIOMPHALARIA ALEXANDRINA SNAIL.

The internal defense system consists of soluble components of hemolymph and circulating cells known as hemocytes. The circulating hemocytes play a central role in innate immunity. This work aimed to study the hemocytes of both susceptible and resistant B. alexandrina snails exposed to S. mansoni infection using light and electron microscopes. Two tested groups were included in the study; 60 susceptible and 60 resistant B. alexandrina snails. Both tested groups were studied as regad the hemocyte count (before and after infection) and the morphological characteristics of both circulating and tissue hemocytes by light and electron microscopes. Before infection, .there was no significant difference between the two groups as regard the hemocyte count, however after infection, there is a significant decrease in the circulating hemocytes of the resistant group. Light microscopy revealed five morphological types of circulating cells of both susceptible and resistant snails. Regarding scannig electron microscopy, hemocytes of susceptible snails appeared rounded with smooth or slightly rough surface. However, that of the resistant snails appeared irregular in shaped with corrugated surface. Furthermore, Light microscopy and the transmission electron microscopy revealed signs of cell activation in the hemocytes of the resistant group. The circulating hemocytes consist of five cell types in both susceptible and resistant B. alexandarina and morphologies of these cells are quite similar, but with more signs of cell activations in the resistant group. More specific studies on the functional activities of the hemocytes and mechanisms that may affect or influence the susceptibility and/or non-susceptibility of molluscs to invade microorganisms is essential and how they can act in the immune response.

COMPARISON BETWEEN KATO-KATZ THICK SMEAR AND SEDIMENTATION TECHNIQUES IN DIAGNOSIS OF FAECAL-ORALLY TRANSMITTED HELMINTHES AND OTHER GEOHELMINTHES.

Kato-Katz technique is widely used for the diagnosis of Fecal-orally transmitted helminthic infections. It is relatively simple and inexpensive. However, a single slide prepared from a single stool specimen has low sensitivity, particularly in light infections. Therefore, there is a great need for concentration techniques. This study detected an accurate and affordable method for diagnosis of fecal-orally transmitted helminthes and other geohelminthes. The study was carried out on 217 stool samples of different sex and age groups. Stool samples were collected from different urban and rural areas in Sharkia Governorate. Stool samples were examined macroscopically and analyzed by different parasitological techniques: direct wet mount (DWM), Kato-Katz thick smear, spontaneous sedimentation in tube technique (SSTT) and formol ether sedimentation technique (FEC). The results showed that formol ether sedimentation method detected 59 positive samples followed by spontaneous sedimentation in tube technique (48/59 positive samples). FEC showed significant difference when compared to direct wet mount and Kato-Katz thick smear. SSTT also showed significant difference when compared to DWM (P <0.05).The overall prevalence of intestinal helminthes was (29.6%) among studied samples in Sharkia Governorate. The commonest helminthic infection was H. nana (12%) followed by E. vemicularis(10%) then A. lumbricoides (3.7%).

ASSESSING THE EFFICACY OF NITAZOXANIDE IN TREATMENT OF CRYPTOSPORIDIOSIS USING PCR EXAMINATION.

Cryptosporidiosis is a gastrointestinal disease of humans and other animals, caused by the genus Cryptosporidium spp. It causes persistent diarrhea and malnutrition and is associated with increased mortality. This study aimed to assess the efficacy of nitazoxanide (NTZ) on clearing the oocysts of C. parvum among infected children using both parasitological and PCR techniques.120 children (1-12y) shedding Cryptosporidium oocysts in their stools were enrolled in the study. They were classified on the basis of the immune status into immunocompetent (ICT) and immunocompromised (ICZ) groups. Each group were subdivided into two groups one of them received'NTZ, and the other received placebo. The efficacy of nitazoxanide was assessed clinically, parasitologically and by nested-PCR technique. At the end of 1st week of treatment, 80% of ICT/ NTZ group and 40% of ICT/ placebo group were free by PCR and 83.3% & 20% respectively were microscopically free. While at the end of 4th week, 93.3% of ICT/NTZ group and 43.3% of ICT/ placebo group were free by PCR and 96.7% & 26.7% respectively were microscopically free. Among the ICZ group, diarrhea was resolved in most patients receiving NTZ within 21 to 28 days of treatment initiation While, it resolved in the ICT group receiving NTZ in most patients within 3 to 5 days of treatment initiation.

Relationship between serum cytokines profiles and hepatic fibrosis in schistosomiasis mansoni: an experimental study.

Forty of eighty mice (10 each group) were infected with S. mansoni cercariae and sacrificed at 3 weeks (G-A), 6 weeks (G-B), 12 weeks (G-C) and 16 weeks (G-D) post infection (P.I). The other forty mice were used as control groups of ten mice each. There were highly significant difference between egg counts after 12 weeks & 16 weeks of infection compared to 6 weeks P.I. The maximum egg count and mature eggs were in 6th week P.I while dead eggs reached the peak at 16th weeks P.I. Liver egg counts showed maximum followed by intestinal and then, stool egg counts. A highly significant differences in hydroxyproline, TGF-Bland IL-4 of infected than in controls and their peak at 16 weeks P.I. A significant difference in the IFN-gamma in the infected than in controls with peak occurred at 6 weeks P.I. and declined after that reaching a low level at 16 weeks P.I. A highly significant positive correlation was between TGF-Bland IL4 and significant negative correlation between IFN-gamma and both IL4 & TGF-B1. A highly significant and significant negative correlation between TGF-B1 and egg count at 12 & 16 weeks P.I respectively. Negative correlation was between IL-4 and egg count at 16 weeks P.I. But, significant positive correlation was between IFN-gamma with the egg count at 16 weeks P.I. A significant negative correlation was between TGF-B1 and oogram at 6 & 16 weeks P.I, but highly significant positivity was between IFN-gamma and oogram at 16 weeks P.I. A significant negative correlation was between IL-4 and oogram at 16 weeks P.I. A significant positive correlation was between levels of hydroxyproline and TGF-B1 at 12 & 16 weeks P.I. Highly significant negative correlation between hydroxyproline and IFN-gamma was at 12 weeks P.I with significant and highly significant positive correlation between hydroxyproline and IL4 at 12 & 16 weeks P.I.