Immunochromatography Lateral Flow Strip Enhancement Based on Passive Gold Nanoparticles Conjugation to Detect Schistosma haematobium Antigens in Human Serum.

PURPOSE: This study aimed to develop and evaluate a lateral flow card for the detection of active Schistosoma haematobium infection. METHODS: In order to prepare the immunochromatography lateral flow strip (ICLFS), antibodies purified from schistosomiasis were conjugated passively with gold nanoparticles using a potassium carbonate buffer. RESULTS: The novel ICLFS was able to correctly identify 64 out of 67 samples of schistosomiasis, 6 out of 90 samples of other parasites, and 0 out of 27 control samples. Sensitivity, specificity, negative predictive value (NPV), and positive predictive value (PPV) were 95.5%, 93.3%, 90%, and 91.4% respectively. Comparatively, the sensitivity, specificity, NPV, and PPV of sandwich enzyme-linked immunosorbent assays (ELISA) conjugated with gold nanoparticles (AuNPs) were 91.1%, 88.8%, 85.9%, and 84.4% respectively. The increased sensitivity and specificity of ICLFS produced superior results to those of sandwich ELISA. CONCLUSION: In conclusion, ICLFS is more beneficial and precise than sandwich ELISA for detection of S. haematobium infection at early stage.

Efficacy of intradialytic amino acids on nutritional status in children with stage 5 chronic kidney disease.

BACKGROUND: Protein energy wasting (PEW) is a common cause of morbidity and mortality in patients with stage 5 chronic kidney disease (CKD 5). Intradialytic parenteral nutrition (IDPN) has been used as a therapy for preventing and treating PEW in children with CKD 5 when other conventional modalities fail. However, not enough data is available to define its effectiveness in treating malnutrition in children. This study aims to investigate potential benefits of IDPN in Egyptian children with CKD 5. METHODS: In this prospective, placebo-controlled, parallel-group single blinded study, we enrolled 50 CKD 5 patients; 25 patients (intervention group) received intravenous amino acids (KIDIMN), while 25 patients (control group) received normal saline as placebo, each given during regular dialysis 3 times a week for 9 months. Patients were subjected to nutritional assessment at baseline and 3-, 6-, and 9-month follow-up using height Z-score, hand grip strength (HGS) for muscle power assessment, body composition monitor (BCM) for assessing lean tissue mass (LTM) and adipose tissue mass (ATM), and biochemical measures including serum albumin, serum triglyceride, and serum cholesterol. RESULTS: When comparing baseline and 9-month follow-up values, significant improvement was recorded in height Z-score, LTM, and serum albumin in the intervention group unlike the control group where no significant changes were recorded. CONCLUSION: IDPN is proposed to be an effective method for preventing and treating malnutrition in children with CKD 5. However, further multi-centric studies with larger sample size and longer duration of follow-up are still recommended.

Partial Capitate with/without Hamate Osteotomy in the Treatment of Kienböck's Disease: Influence of the Stage of the Disease on the Midterm Outcome.

Objective Moritomo et al introduced partial capitate osteotomy as a treatment modality for early stages of Kienböck's disease. This technique maintains articular contact between the capitate and the scaphoid. We added hamate-shortening osteotomy in addition to partial capitate shortening in cases of lunate type II. The purpose of this study was to evaluate intermediate-term results of partial capitate shortening, investigate the influence of the stage of the disease on the outcome, and assess the clinical and radiological outcomes of adding hamate osteotomy in cases of type II lunate. Patients and Methods A total of 17 consecutive patients (3 women, 14 men) with early stages of Kienböck's disease were prospectively reviewed using the aforementioned technique. Eight patients were in stage II and nine patients were in stage IIIA according to the Lichtman classification system. Clinical outcome measures included pain visual analog score, grip strength and range of motion as a percentage of the unaffected side, and assessment using the Patient-Rated Hand and Wrist Evaluation (PRHWE) and the modified Wrightington Hospital Wrist Score (MWHWS). Radiological outcome measures included healing of the osteotomy site, Stahl index, radioscaphoid angle, and progression of the disease. Results Follow-up period averaged 72 months. All cases of isolated capitate osteotomy and combined capitate and hamate osteotomies united fully. Clinical results revealed significant improvement in pain, grip strength and extension, and PRHWE and MWHWS values. Wrist flexion did not change postoperatively. Patients with stage II showed better overall results and significant MWHWS improvement. Conclusion At the intermediate term, partial capitate with/without hamate shortening is an effective modality for the treatment of patients with early stage Kienböck's disease. Stage II patients showed better results than stage IIIA patients in terms of pain, flexion, grip, PRHWE, and MWHWS. Adding hamate osteotomy may improve the functional results for type II lunate; however, a larger sample is needed to elicit statistical significance. Level of Evidence This is a Level IV, therapeutic study.

The Protective Role of Toll-Like Receptor Agonist Monophosphoryl Lipid A Against Vaccinated Murine Schistosomiasis.

PURPOSE: Schistosomiasis is a disease that afflicts over 220 million people worldwide. To date, there is no vaccine against schistosomiasis and chemotherapy relies basically on a single drug, praziquantel. The current study was undertaken to investigate the therapeutic effects of monophosphoryl lipid A (MPLA) as an adjuvant in soluble egg antigen (SEA)-vaccinated and Schistosoma mansoni-infected mice. METHODS: Mice were divided into two groups of uninfected and Schistosoma mansoni infected. The two groups were treated differently with MPLA, SEA and praziquantel. Study parameters included parasitological, immunological and biochemical parameters. RESULTS: Parasitological parameters revealed that intraperitoneal injection of MPLA into SEA-vaccinated and S. mansoni-infected mice was effective in reducing the worm and egg burden, granuloma count and diameter as well as the total area of infection in their livers versus SEA-untreated but infected ones. In addition, MPLA showed ameliorative action on the elevated liver oxidative stress marker, including malondialdehyde (MDA) and a decrease in the level of the antioxidant enzymes, reduced glutathione (GSH) and catalase (CAT) which may have a role in the liver damage and fibrosis due to S. mansoni infection. CONCLUSION: Treatment with MPLA has multi-functions in attenuating the deleterious impacts of S. mansoni infection in mice livers. Its effects are mediated through a reduction of ova count, worm burden, granuloma diameter and amelioration of antioxidant defense systems, and liver function biomarkers.

DNA vaccination using recombinant Schistosoma mansoni fatty acid binding protein (smFABP) gene.

Schistosomiasis is a fatal disease that has a negative impact on health and economics. Praziquantel (PZQ) is the drug of choice for schistosomiasis treatment, but it has no prophylactic effect; therefore, vaccination is an essential requirement in schistosomiasis control. This work was carried out to investigate the possible effect of DNA vaccination against Schistosoma mansoni infection using recombinant S. mansoni fatty acid binding protein (rsmFABP). The smFABP gene was cloned into the eukaryotic expression vector pcDNAI/Amp in order to obtain an smFABP-pcDNAI recombinant plasmid (DNA vaccine) and was used for the intramuscular DNA vaccination of out-bread Swiss albino mice prior to infection with S. mansoni cercariae. Infected groups, either DNA vaccinated or unvaccinated, were treated with PZQ at week 6 post-infection. After 8 weeks post-infection, all mouse groups were sacrificed and parasitological, immunological and histopathological parameters were studied. DNA vaccinated mice showed a high titer of anti-smFABP-IgG antibodies and acquired significant protection (74.2%, p < 0.01) against S. mansoni infection, with a reduction in ova and granuloma counts. DNA vaccinated and PZQ treated animals had higher titers of anti-smFABP-IgG antibodies and decreased (87%, P < 0.001) parenchymal granulomas compared to the DNA vaccinated PZQ untreated group. Infected mice, either non DNA vaccinated or vaccinated, had very high collagen content and fibrous granulomas (74%) compared to the PZQ treated group (10.3% fibrous granuloma) and PZQ treated + DNA vaccinated group (0% fibrous granuloma). In conclusion, DNA vaccination had protective and anti-pathological effects in naive mice and greatly improved the pathological status in PZQ-treated animals, suggesting an immunological and pathological modulating effect of PZQ treatment.

Advantages of bioconjugated silica-coated nanoparticles as an innovative diagnosis for human toxoplasmosis.

Nanotechnology is a promising arena for generating new applications in Medicine. To successfully functionalised nanoparticles for a given biomedical application, a wide range of chemical, physical and biological factors have to be taken into account. Silica-coated nanoparticles, (SiO2NP) exhibit substantial diagnostic activity owing to their large surface to volume ratios and crystallographic surface structure. This work aimed to evaluate the advantage of bioconjugation of SiO2NP with PAb against Toxoplasma lyzate antigen (TLA) as an innovative diagnostic method for human toxoplasmosis. This cross-sectional study included 120 individuals, divided into Group I: 70 patients suspected for Toxoplasma gondii based on the presence of clinical manifestation. Group II: 30 patients harboring other parasites than T. gondii Group III: 20 apparently healthy individuals free from toxoplasmosis and other parasitic infections served as negative control. Detection of circulating Toxoplasma antigen was performed by Sandwich ELISA and Nano-sandwich ELISA on sera and pooled urine of human samples. Using Sandwich ELISA, 10 out of 70 suspected Toxoplasma-infected human serum samples showed false negative and 8 out of 30 of other parasites groups were false positive giving 85.7% sensitivity and 84.0% specificity, while the sensitivity and specificity were 78.6% and 70% respectively in urine samples. Using Nano-Sandwich ELISA, 7 out of 70 suspected Toxoplasma-infected human samples showed false negative results and the sensitivity of the assay was 90.0%, while 4 out of 30 of other parasites groups were false positive giving 92.0% specificity, while the sensitivity and specificity were 82.6% and 80% respectively in urine samples. In conclusion, our data demonstrated that loading SiO2 nanoparticles with pAb increased the sensitivity and specificity of Nano-sandwich ELISA for detection of T.gondii antigens in serum and urine samples, thus active (early) and light infections could be easily detected.

Efficacy of soluble glycoprotein fraction from Allium sativum purified by size exclusion chromatography on murine Schistosomiasis mansoni.

In this work, the efficiency of crude MeOH extracts and soluble glycoprotein fraction of Allium sativum purified by size-exclusion chromatography (SEC) on parasitological, histopathological and some biochemical parameters in Schistosoma mansoni infected mice were investigated. Animals were infected by tail immersion with 100 cercariae/each mouse and divided into five groups in addition to the normal control. The results revealed a significant decrease in mean worm burden in all treated mice especially in the group treated with soluble glycoprotein fraction of A. sativum as compared to infected non-treated control with the disappearance of female worms. Administration of the studied extracts revealed remarkable amelioration in the levels of all the measured parameters in S. mansoni infected mice. In addition, treatment of mice with crude A. sativum MeOH extract and soluble glycoprotein fraction of A. sativum decreased significantly the activities of studied enzymes as compared to the infected untreated group. The highest degrees of enhancement in pathological changes was observed in the treated one with soluble glycoprotein fraction of A. sativum compared to the infected group represented by small sized, late fibro-cellular granuloma, the decrease in cellular constituents and degenerative changes in eggs. In conclusion, A. sativum treatment had effective schistosomicidal activities, through reduction of worm burden and tissue eggs, especially when it was given in purified glycoprotein fraction. Moreover, the soluble glycoprotein fraction of A. sativum largely modulates both the size and the number of granulomas.

Fasciola gigantica fatty acid binding protein (FABP) as a prophylactic agent against Schistosoma mansoni infection in CD1 mice.

Although schistosomicidal drugs and other control measures exist, the advent of an efficacious vaccine remains the most potentially powerful means for controlling this disease. In this study, native fatty acid binding protein (FABP) from Fasciola gigantica was purified from the adult worm's crude extract by saturation with ammonium sulphate followed by separation on DEAE-Sephadex A-50 anion exchange chromatography and gel filtration using Sephacryl HR-100, respectively. CD1 mice were immunized with the purified, native F. gigantica FABP in Freund's adjuvant and challenged subcutaneously with 120 Schistosoma mansoni cercariae. Immunization of CD1 mice with F. gigantica FABP has induced heterologous protection against S. mansoni, evidenced by the significant reduction in mean worm burden (72.3%), liver and intestinal egg counts (81.3% and 80.8%, respectively), and hepatic granuloma counts (42%). Also, it elicited mixed IgG(1)/IgG(2b) immune responses with predominant IgG1 isotype, suggesting that native F. gigantica FABP is mediated by a mixed Th1/Th2 response. However, it failed to induce any significant differences in the oogram pattern or in the mean granuloma diameter. This indicated that native F. gigantica FABP could be a promising vaccine candidate against S. mansoni infection.

Diagnostic efficacy of monoclonal antibody based sandwich enzyme linked immunosorbent assay (ELISA) for detection of Fasciola gigantica excretory/secretory antigens in both serum and stool.

BACKGROUND: This research was carried out to develop a reliable monoclonal antibody (MoAb)-based sandwich enzyme linked immunosorbent assay (ELISA) for the diagnosis of active Fasciola gigantica infection in both serum and stool for comparative purposes. METHODS: From a panel of MoAbs raised against F. gigantica excretory/secretory antigens (ES Ags), a pair (12B/11D/3F and 10A/9D/10G) was chosen due to its high reactivity and strict specificity to F. gigantica antigen by indirect ELISA. RESULTS: The two MoAbs were of the IgG1 and IgG(2a) subclasses, respectively. Using SDS-PAGE and EITB, the selected MoAbs recognized 83, 64, 45 and 26 kDa bands of ES Ags. The lower detection limit of ELISA assay was 3 ng/ml. In stool, the sensitivity, specificity and diagnostic efficacy of ELISA was 96%, 98.2 and 97.1%; while in serum they were 94%, 94.6% and 94.3%, respectively. Moreover, a positive correlation was found between ova count in stool of F. gigantica infected patients and the OD readings of ELISA in both stool and serum samples (r = 0.730, p < 0.01 and r = 0.608; p < 0.01, respectively). CONCLUSIONS: These data showed that the use of MoAb-based sandwich ELISA for the detection of F. gigantica coproantigens in stool specimens was superior to serum samples; it provides a highly efficient, non-invasive technique for the diagnosis of active F. gigantica infection.

Immunological and parasitological parameters after treatment with dexamethasone in murine Schistosoma mansoni.

This work aimed to evaluate the effect of diphenyl dimethyl bicarboxylate (DDB) and dexamethasone alone and in combination with praziquantel on various parasitological, immunological and pathological parameters reflecting disease severity and morbidity in murine schistosomiasis. DDB and dexamethasone had no effect on worm burden but altered tissue egg distribution. This indicates that, under the schedule used, neither drug interfered with the development of adult worms or oviposition, but both can modulate liver pathology. Dexamethasone resulted in a greater reduction in granuloma size than did DDB. Dexamethasone-treated mice also showed lower levels of serum gamma interferon (IFN-γ), interleukin-12 (IL-12) and IL-4, together with higher IL-10 levels, than infected untreated control animals. These data suggest that dexamethasone is a convenient and promising coadjuvant agent that results in decreased morbidity in murine schistosomiasis.

Immunological and parasitological parameters after treatment with dexamethasone in murine Schistosoma mansoni

This work aimed to evaluate the effect of diphenyl dimethyl bicarboxylate (DDB) and dexamethasone alone and in combination with praziquantel on various parasitological, immunological and pathological parameters reflecting disease severity and morbidity in murine schistosomiasis. DDB and dexamethasone had no effect on worm burden but altered tissue egg distribution. This indicates that, under the schedule used, neither drug interfered with the development of adult worms or oviposition, but both can modulate liver pathology. Dexamethasone resulted in a greater reduction in granuloma size than did DDB. Dexamethasone-treated mice also showed lower levels of serum gamma interferon (IFN-γ), interleukin-12 (IL-12) and IL-4, together with higher IL-10 levels, than infected untreated control animals. These data suggest that dexamethasone is a convenient and promising coadjuvant agent that results in decreased morbidity in murine schistosomiasis.

Immunomodulation of pulmonary and hepatic granulomatous response in mice immunized with purified lung-stage schistosomulae antigen.

The present objective was to investigate the possible effect of immunization protocol against Schistosoma mansoni infection using purified lung-stage schistosomulae antigen. Two experimental models (lung & liver) were used, each of 3 groups (Gs): Immunized G. (10 mice) infected control G. (10 mice) and normal control G. (10 mice). Hundred microg of purified schistosomulae antigen followed by two booster doses each of 50 microg antigen and at one week interval were injected intraperitoneally into Swiss albino mice three days prior to intravenous injection of 3000 viable S. mansoni eggs (lung model) or to exposure to 100 cercariae (hepatic model). Mice were sacrificed 16 days post-injection (lung model) and 8 weeks post-infection (hepatic model). Various parasitological parameters, histopathological assessments and immunological parameters were studied. The data revealed that immunization with purified lung-stage schistosomulae antigen induces protective effect against S. mansoni infection. The marked reduction in worm burden, egg load, granuloma diameter and collagen content were accompanied by increased percentage of degenerated ova and amelioration of the associated pathological changes in pulmonary and hepatic tissue. Increased levels of specific immunoglobulins particularly IgG & IgM and decreased ratio of T cell subsets (CD4+/CD8+) in granulomas of both models were also noticed.