Evaluation of schistosomula lung antigen preparation and soluble egg antigen vaccines on experimental schistosomiasis mansoni.

Schistosomiasis causes significant morbidity and mortality worldwide. This study aimed to assess the effect of schistosomula lung antigen preparation (SLAP) and soluble egg antigen (SEA) on a murine schistosomiasis mansoni model. Ninety laboratory-bred male Swiss albino mice were divided into 6 groups. Two doses of the vaccine were given at 2-week intervals. All mice were subcutaneously infected with 80±10 Schistosoma mansoni cercariae 2 weeks after the last vaccination dose. They were sacrificed 7 weeks post-infection. Parasitological and histopathological studies were conducted to assess the effect of inoculated antigens (single or combined). The results showed that the combination of SLAP and SEA (combination group) led to a significant reduction in worm burden (65.56%), and liver and intestine egg count (59% and 60.59%, respectively). The oogram pattern revealed a reduction in immature and mature eggs (15±0.4 and 10±0.8, respectively) and an increased number of dead eggs in the combination group (P<0.001). In terms of histopathological changes, the combination group showed notably small compact fibrocellular egg granuloma and moderate fibrosis in the liver. A high percentage of destroyed ova was observed in the intestine of the combination group. This study demonstrates for the first time the prophylactic effect of combined SLAP and SEA vaccine. The vaccine induced a significant reduction in the parasitological and pathological impacts of schistosomiasis mansoni in hepatic and intestinal tissues, making it a promising vaccine candidate for controlling schistosomiasis.

Evaluating the activity of N-89 as an oral antimalarial drug.

Despite the recent progress in public health measures, malaria remains a troublesome disease that needs to be eradicated. It is essential to develop new antimalarial medications that are reliable and secure. This report evaluated the pharmacokinetics and antimalarial activity of 1,2,6,7-tetraoxaspiro[7.11]nonadecane (N-89) using the rodent malaria parasite Plasmodium berghei in vivo. After a single oral dose (75 mg /kg) of N-89, its pharmacokinetic parameters were measured, and t1/2 was 0.97 h, Tmax was 0.75 h, and bioavailability was 7.01%. A plasma concentration of 8.1 ng/ml of N-89 was maintained for 8 h but could not be detected at 10 h. The dose inhibiting 50% of parasite growth (ED50) and ED90 values of oral N-89 obtained following a 4-day suppressive test were 20 and 40 mg/kg, respectively. Based on the plasma concentration of N-89, we evaluated the antimalarial activity and cure effects of oral N-89 at a dose of 75 mg/kg 3 times daily for 3 consecutive days in mice harboring more than 0.5% parasitemia. In all the N-89- treated groups, the parasites were eliminated on day 5 post-treatment, and all mice recovered without a parasite recurrence for 30 days. Additionally, administering oral N-89 at a low dose of 50 mg/kg was sufficient to cure mice from day 6 without parasite recurrence. This work was the first to investigate the pharmacokinetic characteristics and antimalarial activity of N-89 as an oral drug. In the future, the following steps should be focused on developing N-89 for malaria treatments; its administration schedule and metabolic pathways should be investigated.

Antimalarial effect of synthetic endoperoxide on synchronized Plasmodium chabaudi infected mice.

The discovery of new antimalarial drugs can be developed using asynchronized Plasmodium berghei malaria parasites in vivo in mice. Studies on a particular stage are also required to assess the effectiveness and mode of action of drugs. In this report, we used endoperoxide 6-(1,2,6,7-tetraoxaspiro [7.11] nonadec-4-yl) hexan-1-ol (N-251) as a model antimalarial compound on P. chabaudi parasites. We examined the antimalarial effect of N-251 against ring-stage- and trophozoite-stage-rich P. chabaudi parasites and asynchronized P. berghei parasites using the 4-day suppressive test. The ED50 values were 27, 22, and 22 mg/kg, respectively, and the antimalarial activity of N-251 was verified in both rodent malaria parasites. To assess the stage-specific effect of N-251 in vivo, we evaluated the change of parasitemia and distribution of parasite stages using ring-stage- and trophozoite-stage-rich P. chabaudi parasites with one-day drug administration for one life cycle. We discovered that the parasitemias decreased after 13 and 9 hours post-treatment in the ring-stage- and trophozoite-stage-rich groups, respectively. Additionally, in the ring-stage-rich N-251 treated group, the ring-stage parasites hindered trophozoite parasite development. For the trophozoite-stage-rich N-251 treated group, the distribution of the trophozoite stage was maintained without a change in parasitemia until 9 hours. Because of these findings, it can be concluded that N-251 suppressed the trophozoite stage but not the ring stage. We report for the first time that N-251 specifically suppresses the trophozoite stage using P. chabaudi in mice. The results show that P. chabaudi is a reliable model for the characterization of stage-specific antimalarial effects.

Pioneer Use of Antimalarial Transdermal Combination Therapy in Rodent Malaria Model.

We have previously reported 1,2,6,7-tetraoxaspiro [7.11]nonadecane (N-89) as a promising antimalarial compound. In this study, we evaluated the effect of transdermal therapy (tdt) of N-89 in combination (tdct) with other antimalarials as an application for children. We prepared ointment formulas containing N-89 plus another antimalarial drug, specifically, mefloquine, pyrimethamine, or chloroquine. In a 4-day suppressive test, the ED50 values for N-89 alone or combined with either mefloquine, pyrimethamine, or chloroquine were 18, 3, 0.1, and 3 mg/kg, respectively. Interaction assays revealed that N-89 combination therapy showed a synergistic effect with mefloquine and pyrimethamine, but chloroquine provoked an antagonistic effect. Antimalarial activity and cure effect were compared for single-drug application and combination therapy. Low doses of tdct N-89 (35 mg/kg) combined with mefloquine (4 mg/kg) or pyrimethamine (1 mg/kg) gave an antimalarial effect but not a cure effect. In contrast, with high doses of N-89 (60 mg/kg) combined with mefloquine (8 mg/kg) or pyrimethamine (1 mg/kg), parasites disappeared on day 4 of treatment, and mice were completely cured without any parasite recurrence. Our results indicated that transdermal N-89 with mefloquine and pyrimethamine provides a promising antimalarial form for application to children.

Diagnosis of Toxoplasmosis Using Surface Antigen Grade 1 Detection by ELISA, Nano-Gold ELISA, and PCR in Pregnant Women.

Introduction: The accurate diagnosis of toxoplasmosis has critical importance in pregnant women. Nanotechnology and molecular biology are making possible opportunities for accurate and rapid diagnosis of many infectious diseases. Aim and Methods: The aim of our study was to compare nano-gold ELISA with ELISA and PCR for diagnosis of toxoplasmosis using Toxoplasma surface antigen grade 1 (SAG1) in pregnant women seeking antenatal care in outpatient clinics. Results: PCR showed the highest diagnostic values than nano-gold ELISA and ELISA regarding sensitivity (97.3% versus 89.2% and 83.8%); specificity (100% versus 94% and 88%); and diagnostic accuracy (98.9% versus 91.95% and 86.2%), respectively. There is no statistical difference between PCR and nanogold ELISA results. Discussion: Nano-gold ELISA had a significant improvement in diagnosis than the traditional ELISA method. Most likely with the assistance of nanoparticles, more antibodies enter the antigen-antibody complex because of the considerable improvement in the surface area of nano-gold particles. Conclusion: Although PCR had higher diagnostic values than nano ELISA, nano ELISA is cheaper and easier than PCR. We recommend nano-gold ELISA with SAG1 as a promising technique in the diagnosis of toxoplasmosis and survey studies.

Formulation and evaluation of the antimalarial N-89 as a transdermal drug candidate.

The discovery of new effective and safe antimalarial drugs is mandatory. In this report, we formulate and evaluate transdermal (td) 1,2,6,7-tetraoxaspiro[7.11]nonadecane (N-89) using the Plasmodium berghei rodent malaria parasite in vivo model. The selected solvent for the ointment type of td N-89 was polyethylene glycol (PEG) [PEG400:PEG 4000 = 8:1 (v/w)]. We tested different application areas of 4, 6, and 8 cm2 on the shaved backs of mice. Pharmacokinetic (PK) analysis of N-89 parameters after a single 4 cm2 transdermal application revealed that the Tmax was 2 h, the T1/2 was 1.9 h, and the AUC was 1951.1 ng.h/mL. More than 10 ng/mL of plasma concentration was maintained for 12 h. The ED50 values for the 4, 6, and 8 cm2 application areas in a 4-day suppressive test were 18.9, 25.1, and 26.8 mg/kg, respectively. We additionally tested the cure effect of td N-89 in mice at a dose of 60 mg/kg, twice daily for 4 days at 0.2% parasitemia. Parasites disappeared following day 7 post-treatment in all td N-89 treated groups. Mice were cured without any parasite recurrence or dermal irritation. In conclusion, this study determined for the first time the PK parameters and effect of a new ointment type of td N-89. This suggests that transdermal treatment with N-89 is an effective and safe alternative route for the treatment of malaria, especially in children.

Intestinal Parasite Infections and Accuracy of Direct Thin and Thick Smear, Formol-Ether Sedimentation, Centrifugal Flotation, and Mini-FLOTAC Techniques Among Patients with Gastrointestinal Tract Disorders from the Greater Cairo Region, Egypt.

The aim of the present study was to assess the frequency of intestinal parasitic infection among patients with gastrointestinal tract disorders from the Greater Cairo region, Egypt. In addition, a comparison was made of the accuracy of direct thin and thick smear, formol-ether sedimentation (FEC), centrifugal flotation (CF), and mini-FLOTAC techniques in the diagnosis of infection. Out of 100 patients, the overall prevalence of parasitic infection was 51%. Only 6% had dual infection. Giardia lamblia was the most common parasite (26%), followed by Hymenolepis nana (20%), Entamoeba coli (8%), and Enterobius vermicularis (3%). Except the statistically significant association between E. vermicularis infection and perianal itching and insomnia (P < 0.001), age, gender, and complaints of the examined individuals had no association with prevalence of parasitic infection. Both FEC and CF were equally the most accurate techniques (accuracy = 98.2%, confidence interval [CI] = 0.95-1.0, and κ index = 0.962), whereas the Kato-Katz method was the least accurate (accuracy = 67.5%, CI = 0.57-0.78, and κ index = 0.333). However, mini-FLOTAC-ZnSO4 was the most accurate for diagnosis of helminthic infection, and FEC was more accurate for diagnosis of protozoal infection (accuracy = 100%, CI = 1.0-1.0, and κ index = 1).

Present situation of Schistosoma haematobium infection among primary school aged children in some areas of Qualyobia governorate-Egypt.

The current study determined the present situation of urinary schistosomiasis among primary school children in some areas of Qualyobia governorate in Egypt using different diagnostic methods, and to study the effect of Schistosoma haematobium infection on growth parameters of the affected children.The Results showed that The prevalence rate of S. haematobium infection among school children was 5.3% (32/600-child). The infection was more prevalent in males (7.3%) than females (3.1%). The mean age of children was 9.0 +/- 1.76. All infected children showed hindered growth parameter in comparison to corresponding children (low height, weight and body mass index (BMI) for age Z-score). Water contact activities were more frequent in males than females (P<0.001). The dipstick test specificity was 96.4% (versus 96.7% by microscopic examination) and the sensitivity was 88.2% (versus 76.5% by microscopic examination), which was statistically significant (P<0.001).

A study on biological control of six fresh water snails of medical and veterinary importance.

This study evaluated the molluscicidal effect of Commiphora mnolmol oil extract (Myrrh), on control of six fresh water snails (Lymnaea natalensis, Bulinus truncatus, Biomphalaria alexandrina, Physa acuta, Melania tuberculata and Cleopatra bulimoides). Also, the extract effect on the egg masses of L. natalensis, B. truncatus, B. alexandrina and Ph. acuta was evaluated. Snails and egg masses were exposed at 16-20 degrees C to various concentrations (conc.). LD50 after 24 hours expo-sure were 264/132, 283/195, 230/252, 200/224, 241/246 & 241/246 ppm for young/adult of L. natalensis, B. truncatus, B. alexandrina, Ph. acuta, M. tuberculata and C. bulimnoides respectively. LDtoo after 24 hours exposure were 400/400 for L. natalensis, B. truncatus, B. alexandrina, M. tuberculata and C. bulimoides, and 300/300 for Ph. acuta. Also, complete mortality (100%) was achieved for the egg masses of L. natalensis, B. truncatus, B. alexandrina and Ph. acuta at concentrations of 300, 200, 300 & 400 ppm respectively. Lower concentrations gave the same results after longer exposure. LD100 of C. molmol oil extract (Myrrh) had a rapid lethal effect on the six snail species and their egg masses in high conc. of 300 & 400 ppm. Commiphora molmol is a promising plant to be included with the candidate plant molluscicides. The oil extract of this plant showed a remarkable molluscicidal activity against used snail species.

Helobdella nilae and Alboglossiphonia conjugata leeches as biological agents for snails control.

The efficacy of leeches, as biological agents, in control of snail intermediate hosts of schistosomiasis (Bulinus truncatus, Biomphalaria alexandrina) and fascioliasis (Lymnaea natalensis) as well as their effect on the non-target snails Physa acuta, Melanioides tuberculata and Cleopatra bulimoides was evaluated. Two glossiphoniid snail leeches, Helobdella nilae and Alboglossiphonia conjugata were used. They destroyed egg masses and young snails more rapidly than adult ones. H. nilae showed a stronger destructive effect than A. conjugata. In a descending order, it preferred L. natalensis followed by B. truncatus, B. alexandrina, Ph. acuta, M. tuberculata and lastly C. bulimoides. But, A. conjugata preferred L. natalensis followed by B. truncatus, Ph. acuta, M. tuberculata, B. alexandrina and lastly C. bulimoides. The detailed diagnostic morphology and biology of the two leeches were given.

Preparation of quinoline hexose analogs as novel chloroquine-resistant malaria treatments (1). Synthesis of 4-hydroxyquinoline-beta-glucosides.

Quinoline hexose analogs are expected to be useful as novel agents for treatment of chloroquine-resistant malaria. Here, we report preparation of 4-hydroxy quinoline-beta-glucosides from anilines in 4 steps.

Proteome analysis of new antimalarial endoperoxide against Plasmodium falciparum.

N-89, a new antimalarial endoperoxide, was selected as a promising antimalarial compound showing high activity and selectivity. To study the mechanism of N-89 action, N-89 resistant strain (NRC10) was obtained by intermittent drug pressure. NRC10 had a tenfold increase in the EC(50) value of N-89. No cross-resistance was obtained with other antimalarial compounds. Comparative proteome analysis of N-89 sensitive and NRC10 strains revealed over-expression of 12 spots and down-regulation of 14 spots in NRC10. Fifteen proteins were identified of Plasmodium falciparum origin. The identified proteins representing several functions, mainly related to the glycolytic pathway, and metabolism of protein and lipid. Our results suggest that identified proteins may be candidates of antimalarial endoperoxide targets.