Therapeutic Potential of Mesenchymal Stem Cells on Early and Late Experimental Hepatic Schistosomiasis Model.

Cell-based therapy is emerging as a promising therapeutic approach for a wide range of liver diseases. This study aimed to investigate the regenerative and antifibrotic therapeutic potential of bone marrow-derived mesenchymal stem cells (BM-MSCs) in an early and late experimental hepatic schistosomiasis model. BM-MSCs were isolated from 6-wk-old BALB/c donor male mice, then grown and propagated in culture until cell count was 5-8 × 10(6)/ml. MSCs were then separated and injected into Schistosoma mansoni -infected female BALB/c mice on their 6, 10, 14, and 18 wk post-infection. Mice were sacrificed on the fourth and eighth week after BM-MSCs transplantation in each group. Homing of BM-MSCs was confirmed by PCR detection of male Y-chromosome gene (sry) in the liver tissue of the recipient female mice. The regenerative and antifibrotic potential of BM-MSCs was assessed by histopathological examination, morphometric analysis, electron microscopy, and liver function tests. Schistosoma-infected mice, which were treated with BM-MSCs, showed a decrease in the granuloma size, percentage and density of the fibrotic area, formation of new hepatocytes, and improvement of the liver function tests. Immunohistochemical examination of alpha-smooth muscle actin revealed a significant decrease in the immunoreactive hepatic stellate cells in mice treated with MSCs. Early granulomas (acute infection) showed better response to MSC injection than did later granulomas (chronic infection). Dosing and timing of MSCs transplantation should undergo more investigations in long-term experiments before application to the clinical field. This study is the first to assess and compare the effect of MSCs treatment on early and late granulomas.

Genotyping of Toxoplasma gondii strains from female patients with toxoplasmosis.

Thirty-eight female patients with abortion and intrauterine fetal death were selected from the Obstetric and Gynecology Emergency, Ain Shams University Hospitals, with positive PCR results for toxoplasmosis in previous study. In this study, a rapid and efficient procedure was used for genotyping of T. gondii isolates based on PCR-RFLP assay at SAG2 locus. On the basis of the alleles identified at SAG2 locus, the isolates were grouped into three lineages. Type I was determined by resistance of the 3' & 5' end nested product of the SAG2 locus to cleavage by HhaI & Sau3AI respectively. Resistance of 5' end of SAG2 locus to cleavage by HhaI determined type II. Type III was determined by resistance of the 3' end nested-PCR products of SAG2 locus to cleavage by Sau3AI. Of the 38 isolates, type II was the most prevalent genotype found in 33 (87%). Type I was found in 5 (13%) of the isolates, whereas genotype III was not never found.

Detection of molecular markers of toxoplasmosis among Egyptian patients with miscarriage using avidity IgG-ELISA and Western blotting.

A total of 54 miscarriage patients were divided into 3 groups. GI: 10 toxoplasmosis patients with +ve IgM-ELISA; GII: 24 toxoplasmosis patients with +ve IgG-ELISA, and GIII: 20 non-toxoplasmosis cross-matched females as a control. All groups were subjected to IgG-avidity ELISA & IgG-avidity immunoblotting. Avidity Indices (AI) by ELISA ranged from 22.6% to 73.3% in GI and from 9.6%-75.6% in GII. AI were high (>40%) in 3 (30%) patients in G I and in 8 (33.3%) patients in G II. Sera of GI recognized the 20, 28, 32, 60, 93 & 100 Kda bands with 55% reduction in the 38 and 60 Kda bands after treatment with 6 M urea solutions. Sera of GII recognized the 20, 28, 32, 38, 45, 95-97 & 106 Kda bands. There was 12.5%, 16.6% & 16.7% reduction in the 20, 32, & 106 Kda bands, respectively, after urea. The 38 & 60 Kda bands were identified as good diagnostic markers for the recent toxoplasmosis infection (GI). The 20, 32 & 106 Kda bands were good markers of high avidity antibodies during the chronic toxoplasmosis (GII).

A single-step immunochromatographic lateral-flow assay for detection of Giardia lamblia and Cryptosporidium parvum antigens in human fecal samples.

RIDA Quick immunochromatographic lateral-flow assay was evaluated for diagnosis of giardiasis and cryptosporidiosis as compared to the "gold standard" stool examination. Of the 300 specimens were examined by microscopy of direct wet films, concentrated sediments, modified trichrome and modified Ziehl-Neelsen stained slides, 35 samples of Giardia, ten Cryptosporidium, 35 of other parasites, and 20 negative controls were selected for RIDA Quick test examination. All the samples that gave discrepancy results were retested by the centrifugation prior to preparation for the permanent stained smear. The sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) of RIDA quick test for Giardia were 91.6%, 98.4%, 97% & 95.4% respectively, while that of the microscopic stool examination were 94.5%, 100%, 100% & 96.9%. The sensitivity, specificity, PPV and NPV of RIDA quick test for Cryptosporidium was 91.6%, 100%, 100% & 98.8% respectively, while that of the microscopic stool examination were 83.3%, 100%, 100% & 97.7%.

Characterization of specific Trichomonas vaginalis target antigens from different isolates by immunoblotting against hyperimmune rabbit serum.

The SDS-PAGE and immunoblot methods were used to identify Trichomonas vaginalis specific target antigen(s). Ten T. vaginalis isolates, cultured on TYM media, were analyzed by Sodium Dodecyl Sulphate Poly Acrylamide Gel Electrophoresis (SDS-PAGE), revealed 34 distinct bands. Immunoblotting against a hyperimmune rabbit serum, showed 20 reactive bands ranging from 14-205 kDa. There was isolate to isolate variability among 8 isolates, while 2 isolates (6,7) showed a similar antigenic patterns. Imunoblotting of the isolates showed a total of 20 molecular weight bands, 80% of isolates gave positive immunologic reactions above 100 kDa, while below 100 Kda all the isolates recognized the different molecular weight bands. Out of 20 reacting bands, 5 main bands were detected, 29, 66, 84, 95 & 115 kDa gave positive percent of 60, 60, 60, 90 & 80% among the 10 isolates respectively.

Immunopathogenic role of IgG antibody and RANTES in house dust mite-induced chronic bronchitis.

Based on immunological and clinical examinations, 21 patients were diagnosed as having house dust mite (HDM)-induced chronic bronchitis and classified into three groups according to the clinical presentation of the disease: stable bronchitis, exacerbated bronchitis and asthma on top of bronchitis. Using ELISA, the levels of serum anti-Dermatophagoides farinae and anti-D. pteronyssinus IgG antibodies and plasma RANTES (regulated upon activation, normal T-cell-expressed and secreted; a chemokine with attractive and activator role for eosinophils) were measured in correlation to serum eosinophil cationic protein (ECP, a marker of eosinophil activation and degranulation measured by chemiluminescent immunometric technique). Using immunoblotting, IgG binding components of D. farinae and D. pteronyssinus were determined providing a clue for diagnosis of HDM-induced chronic bronchitis. Significant higher levels of anti-D. farinae and anti-D. pteronyssinus IgG antibodies and RANTES were found in asthmatic group followed by exacerbated chronic bronchitis in comparison to stable bronchitis and control groups. ECP level correlated significantly with IgG and RANTES levels in exacerbated bronchitis and asthmatic groups. The results provided evidence that over expression of IgG and RANTES plays a crucial role, as mediator in immunopathogenesis of HDM-induced chronic bronchitis and as marker of the immunological changes likely responsible for progression of bronchitis to asthma in HDM-sensitive patients yet, RANTES seemed to be an early indicator. Definition of the immunopathogenic role of IgG and RANTES in HDM-induced bronchitis should enable the manipulation of the critical immune response in the hope of establishing new therapies. D. farinae and D. pteronyssinu antigenic bands at > 205 and 205 KDa, respectively, considered together showed 71.4% sensitivity in diagnosis of HDM induced chronic bronchitis and 100% specificity by immuno-blotting.

Avidity IgG: diagnosis of primary Toxoplasma gondii infection by indirect immunofluorescent test.

In the present study, 18 serum samples from toxoplasmosis patients with different clinical manifestations were classified into G1: 9 patients with suspected of having recent T. gondii infection, and G2: 9 patients with suspected of having distant latent infection. Patients with low IgG avidity antibody, 80% of them were positive by IgG avidity-indirect immunofluorescent antibody test (IIFAT) and by IgM-ELISA for toxoplasmosis. However, 20% positive by low IgG avidity- IIFAT, were IgG-ELISA positive for toxoplasmosis. Patients with high IgG avidity antibody, 15.4 % of them were positive by high IgG avidity-IIFAT, and by IgM-ELISA, but 61.5% positive by high IgG avidity-IIFAT, were positive by IgG-ELISA and 23.1% positive by high IgG avidity IIFAT, were positive with both IgM and IgG ELISA. It was concluded that detection of high avidity IgG antibodies by IIFAT excluded recent T. gondii infection, as a confirmatory test and when only a single serum sample is submitted by the patient.

Evaluation of semi-quantitative PCR and IgG & IgM ELISA in diagnosis of toxoplasmosis in females with miscarriage.

One hundred female (age 20-42 yrs) patients were classified into group I: 40 patients presented with abortion in the first trimester; group II: 33 patients with abortion in the second trimester and group III: 27 patients with intrauterine fetal death (IUFD). The positive percentages of semi-quantitative PCR and both IgG & IgM ELISA were 38% and 35% respectively. Ten (26.3%) cases out of 38 were positive for toxoplasmosis by both PCR and ELISA-IgG, while 5 (13.2%) cases out of 38 were positive by both PCR and ELISA-IgM, whereas 16 (42.1%) cases out of 38 PCR positive cases were positive by both ELISA IgG & IgM. Sensitivity and specificity of both ELISA IgG and IgM were 81.57% & 93.54% respectively. False negative by ELISA were found in 7 cases out of 38 positive toxoplasmosis cases detected by semi-quantitative PCR. Three cases out of the 7 cases with false negative by ELISA were detected with a trophozoite copy load of 10(1) trophozoite /mL in the blood sample by semi-quantitative PCR. So, the semi-quantitative PCR detected low levels of parasite DNA recommending its usefulness especially in the early stages of the disease when low amount of antibodies can't be detected by serological method or even by the conventional PCR.

Effect of Ferula assafoetida on experimental murine Schistosoma mansoni infection.

Ferula assafoetida is a hard, resinous, oily herbaceous gum belonging to plant family Umbelliferae. It is used as a traditional medicine in many parts of the world and its wide use in medicine was listed by many authors. In the current study, the effect of F. assafoetida on Schistosoma mansoni in experimentally infected mice is investigated. F. assafoetida was given orally via intragastric tube in an oil-form and a powder-form in different concentrations. Four test groups of 30 mice each were studied. Gs I & II mice were given F. assafoetida in an oil-form in different concentrations at 4 and 6 weeks post infection (PI) respectively. Mice in Gs III & IV were given re-constituted F. assafoetida powder in different concentrations (conc.) at 4 & 6 weeks PI. respectively like the previous groups. Oil-form F. assafoetida was given at conc. of 50, 25 & 15 mg/ml. Powder-form was given at conc. of 32, 16 & 8 mg/ml. A highly significant statistical difference between the test Gs (I, II, III & IV) was recorded in comparison to the infection control G with p value < 0.0001 and also between powder and oil forms of F. assafoetida (p < 0.0001) as regards the mean worm burden and tissue egg count. The highest reduction in worm burden and egg counts was found with powder form of F. assafoetida (Gs III & IV) when compared to oil form (Gs I & II), as confirmed histopathologically and by ultrastructural profile alteration.