Immunohistochemical aspects of immunological cross-reaction and masking of epitopes for localization studies on pregnancy-associated plasma protein A

Summary The influence of antibody absorption procedures and proteolytic pre-treatment of formaldehyde-fixed placental tissue on the localization of pregnancy-associated plasma protein A by immunoperoxidase technique was examined.Apparently monospecific IgG fraction of the anti-plasma protein applied directly on fixed tissue resulted in staining of connective tissue and a thin apical rim of the syncytiotrophoblast. Further absorption of the antibody with foetal connective tissue abolished this staining reaction. Pre-treatment of the fixed placental tissue with trypsin prior to application of the antibody, which had been absorbed with connective tissue, resulted in staining within the cytoplasm of the syncytiotrophoblast exclusively. Identical staining was seen when this IgG preparation was used directly on frozen placental tissue.The results point to the importance of the specificity of the antibody preparations and of proteolytic unmasking of epitopes when fixed tissues are used for localization studies of pregnancy-associated plasma protein A by immunoperoxidase technique.     

FAMILIAL VALVULAR PULMONIC STENOSIS INVOLVING THREE SIBLINGS

This report describes the cases of three mentally and physically well-developed siblings, 12, 10, and 8 years of age, with varying degrees of isolated valvular pulmonic stenosis not related to age. The severest lesion occurred in the middle patient and was associated with a right-to-left shunt through a patent foramen ovale. The three children had no other siblings, and there was no history of congenital heart disease among close relatives. These cases support the conclusion that genetic factors play a significant role in the development of certain congenital cardiac disorders.     

Spectrofluorimetric determination of eptifibatide in human plasma and dosage form

A spectrofluorimetric method for the determination of eptifibatide is presented based on its native fluorescence. The type of solvent and the wavelength of maximum excitation and emission were carefully selected to optimize the experimental conditions. Under the specified experimental conditions, the linearities obtained between the emission intensity and the corresponding concentrations of eptifibatide were in the range 0.1–2.5 μg/ml for the calibration curve constructed for direct determination of eptifibatide in dosage form and 0.05–2.2 μg/ml for the calibration curve constructed in spiked human plasma with a good correlation coefficient (r > 0.99). The lower limit of quantification for the calibration curve constructed in human plasma was 0.05 μg/ml. Recovery results for eptifibatide in spiked plasma samples and in dosage form, represented as mean ± % RSD, were 95.17 ± 1.94 and 100.29 ± 1.33 respectively. The suggested procedures were validated according to the International Conference on Harmonization (ICH) guidelines for the direct determination of eptifibatide in its pure form and dosage form and United States Food and Drug Administration (US FDA) Guidance for Industry, Bioanalytical Method Validation for the assay of eptifibatide in human plasma.     

Identification and re-addressing of a transcriptionally permissive locus in the porcine genome

Recently, we established the Sleeping Beauty transposon system for germ line competent transgenesis in the pig. Here, we extend this approach to re-target a transposon-tagged locus for a site-specific gene knock-in, and generated a syngeneic cohort of piglets carrying either the original transposon or the re-targeted event. A Cre-loxP-mediated cassette exchange of the tagging transposon with a different reporter gene was performed, followed by flow cytometric sorting and somatic cell nuclear transfer of recombined cells. In parallel, the original cells were employed in somatic cell nuclear transfer to generate clone siblings, thereby resulting in a clone cohort of piglets carrying different reporter transposons at an identical chromosomal location. Importantly, this strategy supersedes the need for an antibiotic selection marker. This approach expands the arsenal of genome engineering technologies in domestic animals, and will facilitate the development of large animal models for human diseases. Potentially, the syngeneic cohort of pigs will be instrumental for vital tracking of transplanted cells in pre-clinical assessments of novel cell therapies.     

Nucleic Acid Related Compounds: A Convenient Synthesis of 3-Deazauridine Analogues

Abstract

A novel synthesis of 3-deazapyrimidine glycosides utilizing pyridine-2(1H)-ones or their sodium salts and α-halomonosugars namely D-glucose or D-galactose as starting components is described.     

Nutrient sequestration potential of water primrose Ludwigia stolinefera (Guill. & Perr.) P.H. Raven: A strategy for restoring wetland eutrophication

The current work investigates the capacity of the water primrose (Ludwigia stolinefera) to sequester inorganic and organic nutrients in its biomass to restore eutrophic wetlands, besides its nutritive quality as fodder for animals. The nutrient elements and nutritive value of the water primrose were assessed seasonally in polluted and unpolluted watercourses. The water primrose plants’ highest biomass was attained during summer; then, it was significantly reduced till it reached its lowest value during winter. In the polluted canal, the plant root and shoot accumulated higher contents of all nutrient elements (except Na and Mg) rather than in the unpolluted Nile. They accumulated most investigated nutrients in the growing season during summer. The shoots accumulated higher contents of N, P, Ca, and Mg than the root, which accumulated higher concentrations of Na and K. Therefore, summer season is the ideal time to harvest water primrose for removing the maximum nutrients for restoring eutrophic watercourses. The aboveground tissues had the highest values of ether extract (EE) during spring and the highest crude fibers (CF) and total proteins (TP) during summer. In contrast, the belowground tissues had the lowest EE, CF, and TP during winter. In spring, autumn, and winter seasons, the protein content in the grazeable parts (shoots) of the water primrose was within the range, while in summer, it was higher than the minimum requirement for the maintenance of animals. There was a decrease in crude fibers and total proteins, while an increase in soluble carbohydrates content in the below- and above-ground tissues of water primrose under pollution stress. The total protein, lipids, and crude fibers of the aboveground parts of water primrose support this plant as a rough forage.     

Synthesis and Anti-HIV Activity of Different Novel Nonclassical Nucleosides

Abstract

A series of different novel nonclassical nucleosides have been synthesised and evaluated for their inhibitory activity against human immunodeficiency virus (HIV) replication in MT-4 cells.     

Growth and nutrients accumulation potentials of giant reed (Arundo donax L.) in different habitats in Egypt

Arundo donax L. has a high biomass production and a tendency toward community dominance in many habitats and thereby a tolerance to a wide range of environmental conditions. Therefore, the present study investigated the potentiality of A. donax to accumulate nutrients and trace metals in its biomass. Six main habitats (Nile Bank, Drain Bank, Canal Bank, Field Edges, Railways and Roadsides) were recognized. At each habitat, six quadrats (each 1 m²), distributed equally in two sites, were selected for growth measurements (e.g., density, shoot height, diameter, leaf area and biomass), plant and soil analyses. Plants from Nile, Canal and Drain Banks had the highest values of most growth measurements, while those from Railways and Roadsides had the lowest. Canal Bank plants accumulated the highest concentrations of P, Cu and Pb in their leaves; Zn in the stem; and Mg, Cd and Fe in the rhizome. The bioaccumulation factor (BF) of A. donax, for Cd, Fe, Mn and Zn, was greater than 1, while the translocation factor (TF) of most trace metals was less than unity in most habitats. In conclusion, A. donax showed morphological plasticity in response to habitat heterogeneity, and its growth was most vigorous in the riparian habitats. The high BF, as well as the significant positive correlations between trace metals, especially Cd, in soil and plant, renders A. donax a powerful phytoremediator.