Evaluation of Echinostoma liei worm, metacercaria and redia antigens for schistosomiasis control.

While chemotherepeutic drugs, such as praziquantel, oxamniquine and metrifonate, are currently considered safe and effective drugs for schistosomiasis treatment, reinfection occurs frequently after drug treatment. Thus, a vaccine is sought to provide long-term treatment. Antigens from worm, metacercaria and redia of Echinostoma liei (E. liei) were purified using CNBr-activated Sepharose column, then used for immunization of mice prior to infection with Schistosomiasis mansoni. Worm burden, hepatic and intestinal eggs and oogram count was significantly reduced and that was reflected in normalization of liver architecture. This referred to a significant increase in the tested immunoglobulin level (IgM, IgG1 and IgG2).

Evaluation of purified 27.5 kDa protoscolex antigen-based ELISA for the detection of circulating antigens and antibodies in sheep and human hydatidosis.

Hydatidosis is a zoonotic disease caused by the larval stage of Echinococcus granulosus, and the diagnosis of hydatidosis to date remains unresolved despite the development of many serological techniques. The present study aimed to develop an antigen-based enzyme-linked immunosorbent assay (ELISA) using IgG anti-27.5 kDa protoscolex antigen (27.5 PA) for measuring circulating protoscolex antigen (CPA), for comparison with an antibody detection assay, in sera of naturally infected sheep and humans in highly endemic areas in Egypt. In sheep, the sensitivity of ELISA in detecting anti-27.5 PA IgG and CPA was 75.0 and 60.0%, respectively, and the recorded specificity was 80.0 and 88.0%, respectively. In humans, the sensitivity of ELISA in detecting anti-27.5 PA IgG and CPA was 62.5 and 52.5%, respectively, while the specificity of the assay was 66.7 and 75.0%, respectively. In conclusion, an antibody detection assay is still superior and is more sensitive than an antigen detection assay, especially in diagnosing an active infection in which hydatid cysts are predominant. An antigen detection assay may be a useful approach to assessment of the efficacy of treatment, especially after removal of the cyst. Further studies are recommended to improve the diagnostic efficacy of an antigen-based ELISA method by using a highly purified recombinant antigen.

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.

Fasciola gigantica excretory-secretory products for immunodiagnosis and prevention of sheep fasciolosis.

Excretory-secretory products (ESP) products of ex vivo Fasciola gigantica adult worms were used for immunodiagnosis of sheep experimental infection with F. gigantica and natural infection with Fasciola spp. by enzyme-linked immunosorbent assay (ELISA) and Western blotting. Specific IgG antibody binding to native or denatured ESP was detected as early as 2 weeks after experimental sheep infection with 100 or 200 metacercariae. No specific IgG antibody binding was displayed by sera obtained from 192 sheep considered to be Fasciola- and other parasite-free by microscopic examination of bile and feces. Additionally, sera from 200 apparently Fasciola-free sheep, yet infected with other parasites, were all negative. The data, thus, indicated that ESP-based ELISA reached nearly 100% sensitivity and specificity in immunodiagnosis of sheep fasciolosis. As expected, the ESP molecules were immunogenic in sheep eliciting interleukin-12p40 mRNA response and considerable amounts of antibodies, which were able to bind to the surface of newly excysted juvenile worms as judged by membrane indirect immunofluorescence, and mediate their attrition via antibody-dependent cell-mediated cytotoxicity. The ESP-induced cellular and humoral immune responses were associated with a modest reduction in worm count, yet with a highly significant (P<0.0001) decrease in size of recovered worms, thus suggesting that ESP immunization might be a safe and cost-effective strategy for reducing transmission of the infection.

DOWN REGULATION OF CLASSICAL MONOCYTES SUBSET IN PATIENTS WITH HCV RELATED LIVER FIBROSIS.

Chronic liver disease is a worldwide common pathology characterized by inflammatory and fibrotic processes that may lead to progressive evolution from chronic hepatitis to cirrhosis. Peripheral blood monocytes may play an important role in the pathogenesis and resolution of liver fibrosis. These cells may offer new approaches for better understanding the pathogenesisof liver fibrosis. This work defined the proportion of circulating classical monocyte subset with hematopoietic origin in peripheral blood to establish the possible potential role of this subset as non-invasive biomarker of liver fibrosis in patients with HCV related chronic liver disease. Forty patients with HCV induced chronic liver disease were classified according to the stage of liver fibrosis after METAVIR score into 4 groups, patients with stages F1, F2, F3 & F4 liver fibrosis (10 patients each) and 10 healthy subjects served as normal controls. Flowcytometric analysis for immunophenotypic characterization for identification of levels of circulating peripheral blood classical monocytes subset in different groups studied was carried out using monoclonal antibodies anti-CD45, anti-CD 14 and anti-CD 16. THE RESULTS: data demonstrated a significant down regulation (p< 0.01) in the proportion of classical monocytes subset (CD45+ve, CD 14+ve and CD 16-ve) in patients with chronic hepatitis C related liver disease compared to healthy subjects. Data also demonstrate that down regulation of the expression of classical monocyte subset paralleled the worsening severity of liver disease and the progression of liver fibrosis.

Infection with Plasmodium chabaudi diminishes plasma immune complexes and ameliorates the histopathological alterations in different organs of female BWF1 lupus mice.

OBJECTIVE: To determine the effect of Plasmodium chabaudi infection on the plasma level of circulating immune complexes (CICs), haemoglobin (Hb) content, urine profile, and histological features of female BWF1 mice, the murine model of systemic lupus erythematosus (SLE). MATERIALS AND METHODS: A total of 30 female BWF1 lupus mice were randomly divided into three groups as follows: group (I) control group (P. chabaudi uninfected); group (II) lupus mice infected with live P. chabaudi; group (III) lupus mice infected with irradiated P. chabaudi. Urine samples were daily collected from the second week-post infection. Mice from the three groups were killed at day 14 post-infection and heparinized blood was collected for further haemoglobin contents and plasma analysis. Paraffin-embedded kidney, liver, lung, heart, brain, ovary and skin tissues were stained with Hematoxylin and Eosin (H&E) and examined under light microscope. RESULTS: Our results reveal that infection of lupus mice with live P. chabaudi was associated with an increase in urinary Hb and a decrease in plasma Hb and CIC levels. Interestingly, infection of lupus mice with live P. chabaudi ameliorates the histopathological alterations mediated by lupus disease in kidney tissues. Although no parasite sequestration was observed in any of the investigated organs, P. chabaudi pigment deposition was observed in the liver of both live and irradiated P. chabaudi infected groups. CONCLUSIONS: This study in lupus prone BWF1 mice indicated that gamma-irradiated P. chabaudi infection has the desired lupus ameliorating effect without negative effects of malaria which assist the understanding of different responses to plasmodium sp. infection in human lupus patients.

Serological versus antigen detection methods for Giardia duodenalis diagnosis.

Giardiasis constitutes an important public health problem in the world. Contamination of the water with fecal materials including viruses and pathogenic protozoa still represents an environmental health hazard, especially in rural areas. The survey study evaluated the relation between seropositivity and some risk factors. Moreover, the study compared between the serological IgG and IgM level and antigen detection methods for the diagnosis of giardiasis. The results indicate that sex distribution and age were the mean risk factors for seroprevelence. In this study, sera samples were employed in sandwich ELISA assay, to detect circulating Giardia antigens. None of the negative control serum samples gave a positive reaction, but cross reaction was encountered with 3 case of Cryptosporidium. The specificity of the assay was 94.830/a. On the other hand, the sensitivity of the Giardia patient's sera was 94.12% which was higher than that of IgG (86.25%) and IgM (87.50%) secretion measurements. In conclusion, antigen detection methods give better and earlier diagnosis for giardiasis can be performed quickly and do not require an experienced and skilled morphologist.

Protection against Fasciola gigantica using paramyosin antigen as a candidate for vaccine production.

Yet no vaccine to protect ruminants against liver fluke infection has been commercialized. In an attempt to develop a suitable vaccine against Fasciola gigantica (F. gigantica) infection in rabbits, using 97 kDa Pmy antigen. It was found that, the mean worm burdens and bile egg count after challenge were reduced significantly by 58.40 and 61.40%, respectively. On the other hand, immunization of rabbits with Pmy induced a significant expression of humoral antibodies (IgM, total IgG, IgG1, IgG2 and IgG4) and different cytokines (IL-6, IL-10, L-12 and TNF-alpha). Among Ig isotypes, IgG2 and IgG4 were most dominant Post-infection (PI) while, recording a low IgG1 level. The dominance of IgG2 and IgG4 suggested late T helper1 (Th1) involvement in rabbit's cellular response. While, the low IgG1 level suggested Th2 response to adult F. gigantica worm Pmy. Among all cytokines, IL-10 was the highest in rabbits immunized with Pmy PI suggesting also the enhancement of Th2 response. It was clear that the native F. gigantica Pmy is considered as a relevant candidate for vaccination against fascioliasis. Also, these data suggested the immunoprophylactic effect of the native F. gigantica Pmy which is mediated by a mixed Th1/Th2 response.

Overview on cysteine protease inhibitors as chemotherapy for Schistosomiasis mansoni in mice and also its effect on the parasitological and immunological profile.

The present study evaluated the use of 3 types of Cysteine Protease Inhibitors (CPIs) with praziquantel (PZQ) as chemotherapy against schistosomiasis mansoni in mice. All groups were going to assessment of fluromethylketone (FMK), Vinyl Sulfone (VS) and Sodium Nitro Prussid (SNP) by measurement of parasitological, immunological and histological parameters. In our study, The ova count/gm liver or intestine on with PZQ treatment showed 99.1 and 95.2% Percent Reduction (PR), respectively compared to control group. The most effective CPI was FMK when combined with PZQ recording 99.8 and 99.6% PR for liver and intestine, respectively. Regarding to the oogram pattern, FMK, VS and SNP treatment either at 3 or 5 wk PI revealed marked decrease in the immature and mature ova counts and an increase of the dead ova percentages. The effect of CPIs was studied on the PR of Mean Granuloma Diameter (MGD) and Mean Granuloma Number (MGN) of infected treated groups compared to infected control and PZQ treated groups. FMK treatment proved to be highly was effective against S. mansoni in mice disintegrating ova and reduction in granulomatous size and numbers. The microscopic examination of liver sections of infected mice showed a large cellular granuloma with living central ova. sections of Infected mice liver treated with FMK or VS alone or combined with PZQ showed a great reduction in granuloma size as small cellular granuloma with central degenerated ova. We observed that these CPIs alone or combined with PZQ could effectively block schistosomal activity and prevented its growth and differentiation. Briefly, the best schistosomicidal effect of CPIs, that gained by drug administration orally in a dose of 50 mg kg(-1) mouse, was observed with FMK. This was followed by VS and lastly with SNP. These results gave evidence that CPIs can selectively arrest parasite replication without untoward toxicity to the host.

Trends in rates of retinal surgery in England from 1968 to 2004: studies of hospital statistics.

AIM: To describe trends over time and geographical variation in rates of vitreo-retinal surgery in England from 1968 to 2004. METHODS: Routinely collected hospital statistics were analysed for England, using the Hospital In-patient Enquiry and Hospital Episode Statistics from 1968 to 2004, and for the Oxford National Health Service Region using the Oxford Record Linkage Study from 1963 to 2004. RESULTS: Annual admission rates in England for surgery were about 5 episodes per 100 000 population in the 1960s, rising gradually to about 10 per 100 000 in the early 1990s, and then more sharply to 30 by 2004. The Oxford Record Linkage Study shows that multiple admissions per person were rare, which confirms that the observed increase represents a real increase in the number of people treated. Annual rates in England for buckle procedures declined to about 6 episodes per 100 000 population in 2004, whereas vitrectomy surgery rose to about 26 episodes per 100 000 population in 2004. CONCLUSION: Vitreo-retinal surgery has developed over the last 40 years in England, rapidly so over the past 15 years. Vitrectomy surgery has become much more common in England and buckle surgery has shown a steady decline. We demonstrate statistically significant geographical variation in the current annual rate of surgery between local authorities in England.

Identification and control of nosocomial adenovirus keratoconjunctivitis in an ophthalmic department.

AIMS: To identify the extent of nosocomial adenovirus keratoconjunctivitis (AKC) and assess the effect of a new infection-control policy. METHODS: Nosocomial AKC was defined as AKC in patients attending the hospital within 3 weeks of a previous visit for an unrelated non-infective condition. An audit of culture-proven nosocomial AKC was carried out from October 1998 to September 1999 to establish its extent in our outpatient department. A new infection-control policy for AKC was introduced in June 2001 that differed from the previous policy by segregating suspected AKC cases in a separate waiting area and examination room, and by expediting their examination, to reduce their exposure to both staff and patients in the common waiting areas. In October 2002, AKC was made an index infection and subjected to continual quarterly audit; the figures for this until December 2005 are reported. RESULTS: In the 1998-9 audit, before the introduction of the new infection-control measures, 48.4% (75/155) AKC infections were nosocomial. In the 12 months following the introduction of the quarterly audit in October 2002, 22.7% (5/22) were nosocomial, but thereafter the numbers of nosocomial cases dropped to 3.4% (8/235). CONCLUSION: Introduction of audit of nosocomial AKC infection identified that there was chronic cross-infection in the Moorfields Eye Hospital Accident and Emergency Department. This was controlled by the introduction of patient segregation, as an additional infection-control measure, which has almost eliminated nosocomial AKC transmission in the hospital.