Immunological identification of fasciola hepatica antigens containing major human T - cell and B - cell epitapes

Fasciola hepatica whole worm homogenate [Fhwwh] separated fractions were used in enzyme linked immunoelectro-transfer blot [EITB] to identify the antigen[s] which induce antibody formation in human fascioliasis. The immuno-reactive antigens recognized by the infected patients were 25-29 kDa and 12 kDa. The antigens were biochemically purified by model 491-prep cell fraction [BIO-RAD]. The capability of the purified antigens to induce humoral and cellular responses with cells and sera of infected patients was investigated using enzyme linked immunosorbent assay [ELISA] and lymphoproliferative responses techniques. The 25-29 kDa cluster of the antigen[s] were found to be more efficient in inducing lymphoproliferative response than 12 kDa; thus, it was considered as the target antigens used in the generation of human monospecific polyclonal immunopurified antibody probes [IPAb]. The specificity and immuno-reactivity of the IPAb with Fhwwh, F. hepatica excretory secretory products [FhESP] and S. mansoni adult worm antigen [SAWA] were evaluated by using EITB. The results showed that IPAb was immuno-reactive with 25-29 and 12 kDa antigens of both Fhwwh and FhESP

Cloning and sequence analysis of genes encoding Fasciola hepatica immunodominant antigens

An immuno-screening of fusion protein produced in lambda plaques was done using IPAb. Two clones were isolated and identified as Fh lambda 400 and Fh lambda 800. Both clones were sequenced, Fh lambda 400 contained 305 translated bases encoding 11.509 kDa and designated as SFh12, while Fh lambda 800 contained 311 translated bases encoding protein of 11.058 kDa designated as SFh11. The DNA sequence homology search of Fh lambda 400 revealed a relatively high degree of identity with F. hepatica amoebapore-like protein mRNA. However, Fh lambda 800 revealed the highest similarity with F. hepatica tegumental antigen [T1] mRNA. The protein homology search of SFh12 gave 100% identity with amoebapore-like protein [APLP], while SFh11 showed 75% identity with F. hepatica tegumental antigen [TA]. The biochemical analysis of the deduced proteins was identified. In addition, the predicted T- and B- cell epitopes were also evaluated. However, a histological localization of the identified antigens was achieved using the IPAb in an indirect immunofluorescent antibody assay [FA]. The results revealed that the IPAb labeled the outer glycocalyx in a characteristic pattern, which proved that the identified antigens were tegumental in origin and the infected fasciola subjects induced antibodies directed mainly against tegumental components

Allele profiles of short tandem repeat loci in Egyptians

In this study, relative allele frequencies of 4 short tandem repeats [STR] loci in 47 unrelated subjects from Egyptian population as a primary step for forming a nationwide database was reported. The sample represented various geographical, ethnic and religious backgrounds that constituted contemporary Egyptian population. DNA was extracted from whole blood using salting out method. The allele patterns in Egyptian sample showed high levels of heterozygosity over 74% in 3 loci. The interpopulation differences were least detected with Caucasians and most with Asian and Hispanic Americans

Cytogenetic effect of consecutive doses of a constant magnetic field I-induction of micronuclei in bone-marrow cells in mice

The cytogenetic effect of a constant magnetic field [400G] was studied with the micronucleus test in mice bone marrow. Adult male mice were exposed to 5 consecutive doses, each separated by 24-hour intervals, for 30 minutes. All doses induced a significant increase in the number of micronucleated polychromatic erythrocytes [PCE]. The highest micronuclei were observed after exposure to double, while the lowest micronuclei were after pentad dose. Moreover, the dose response effect showed that the increase in micronuclei was linearly up to double dose and, thereafter, the increases dropped and slightly deviated from linearity. In conclusion, the clastogenic and/or aneugenic effects, observed in this study, of magnetic field may be closely correlated with cell transformation, tumorigenesis and aneuploidy

Mutagenic effect of rift valley fever virus as measured by indication of chromosomal aberrations and formation of micronuclei in mice bone marrow cells

The mutagenic effect of Rift Valley fever [RVF] virus was tested in male mice bone marrow cells. Three virus titers were chosen [low [L], intermediate [IM] and high [H]]. These were 3, 300 and 3000 VP/dose, respectively. Mice were inoculated with each titer and samples were taken 24 and 48 hours later from bone marrow. An increase in chromosomal aberrations [structural and numerical] was observed in some treatments with RVF virus. The significant structural chromosomal aberrations are in the form of gape and centromeric attenuations, while the numerical aberration was endomitosis. All treatments showed a decrease in mitotic index [number of dividing nuclei per 300 cells], this decrease was highly significant. Although, the number of micronucleated PCE increased after inoculation with all treatments, the increase was below the significant level. Since there is no information available on the mutagenicity of RVF virus, more cytogenetic studies are suggested, the result of which could be compared with these results. This may throw some light on the mutagenic effect that could be produced by RVF virus