Screening of diseases associated with abnormal metabolites for evaluation of HPLC in organic aciduria profiling

Organic acid disorders are a heterogeneous group of inborn errors of metabolism, in which organic acids accumulate in the body. They have high prevalence in Egypt because of a high rate of consanguineous marriages. Here we report our experience with the diagnostic evaluation of patients with organic acidemias as well as several other inborn errors of metabolism [lEMs] by liquid chromatography-tandem mass spectrometry [LC/MS-MS], gas-chro-matography mass spectrometry [GC/MS] and by isocratic cation exchange [high-performance liquid-chromatography] [HPLC] to evaluate the use of HPLC method for disease-associated metabolite screening. In this study, we screened 86 suspected Egyptians patients with organic acid disorders by LC/MS-MS, GC/MS and by HPLC aged from 3 days to 12 years old. Data obtained from the three methods were statistically analyzed to evaluate the specificity and sensitivity of the HPLC method over the other two methods and to pursue its precision in the diagnosis of organic acid disorders. Moreover, 17 urine samples were collected from patients with several other lEMs to evaluate the efficiency of HPLC in detecting abnormal metabolites in urine samples. The screening results showed that diagnostic efficiencies were varied among the three methods, HPLC showing a higher sensitivity of detecting normal urine as well as a highly satisfactory extent for the detection of different metabolic disorders. In addition, some typical urinary HPLC chromatograms of different metabolic disorders were presented to help the investigator who is going to start an organic aciduria screening program by HPLC to be familiar with various patterns. This study has indicated that HPLC is an easy applicable and useful technique for the initial screening of organic acid disorders and many other disease associated metabolites

Identification of cDNA clones ending for antigens of Schistosoma mansoni schistosomula

In this study, cDNA clones coding for a number of surface antigens were identified by immunoscreening S. mansoni sporocyst cDNA library with highly protective antibodies derived from sera of rabbits vaccinated with irradiated cercariae. They were prepared by an affinity purifying the sera over column of Sepharose-4B coupled to NP-40 extracts of 3-hour schistosomula. The antibodies displayed a strong reactivity with NP-40 extracts of 3-hour schistosomula when tested by enzyme linked immunosorbent assay as well as strong as affinity for the surface of schistosomula as determined by indirect immunofluorescence assay. Three different clones coded for three different types of enzymes were isolated and identified [serine proteinase inhibitor [SPN], mitochondria ATP synthase [ATPase] and lysozyme]. The isolation of those clones allowed for antigen characterization and expression for vaccine trials