Optimization of the cost and sensitivity of receptor- and enzyme-based assays.

In detecting receptor antagonists or enzyme inhibitors, there are three parameters that often affect the outcome in a predictable quantitative manner: concentrations of the receptors (enzyme), labeled ligand (substrate), and antagonist (inhibitor). The usual goal of assay optimization is to maximize the ability of the assay to detect low concentrations of the analyte. Another question of practical importance, especially in screening of large numbers of samples, would be minimization of the reagent cost. Although the mathematical theory of optimization of the receptor binding assay was developed a long time ago, the resulting formulas (in the general case of unequal affinities of ligand and competitor) were not well suited for practical use. The current availability of computational programs, such as Mathematica, makes possible an efficient solution, both for receptor- and enzyme-based assays. We use a graphical approach to assay optimization and apply it to the following problems: (1) optimization of assay sensitivity, (2) optimization of the reagent cost, and (3) analysis of the entire range of the parameter values since the mathematically optimal values may sometimes be impractical. The computation is extremely simple and the problem can sometimes be solved in several minutes.     

Distribution of R-r-i-blood group system in Australian sheep

The distribution of the R-r-i blood group system was studied in eight breeds of Australian sheep. The recessive i gene was found to be absent from Merinos and, consequently, the i phenotype was absent in Merino crosses. Distribution of the groups in Dorset Horns and Ryelands was found to be strikingly different from that of other breeds. Two animals were found to have anti-r antibodies in serum.     

Intestinal brush border and lysosomal enzymes and immunoglobulin absorption in the newly-born lamb.

Activities of the brush border enzymes alkaline phosphates, leucine aminopeptidase and lactase and the lysosomal enzymes alpha-mannosidase and beta-N-acetylglucosaminidase increased in the serum of newly-born lambs fed colostrum. Feeding lipid and protein components of colostrum and bovine serum resulted in enzyme responses similar to those observed after feeding colostrum. Activities of each of the enzymes increased in mesenteric lymph collected from newly-born lambs when immunoglobulins were being absorbed from the jejunum and ileum.     

Leukotriene D4-induced increases in the cytoplasmic pH of human myelocytic leukocytes

The effects of leukotriene D4 on the intracellular pH of human myelocytes, derived from cultured HL-60 cells by dimethylsulfoxide-induced differentiation, were quantified with the fluorescent indicator 2',7'-bis-(2-carboxy-ethyl)-5,6-carboxyfluorescein. Leukotriene D4, but not C4 or E4, increased intracellular pH optimally by 3 min with a half-maximal effect at 1-2 nM. The increases in intracellular pH stimulated by leukotriene D4 were prevented by pretreatment of myelocytes with leukotriene D4 but not peptide chemotactic factors. Analogs of amiloride that inhibit selectively the Na+/H+ antiport also prevented the intracellular alkalinization induced by leukotriene D4. The rate of recovery of intracellular pH after an acid load with 30 mM sodium propionate was approximately 30% higher at each level of intracellular pH for myelocytes exposed to leukotriene D4 than for those challenged in buffer alone. The increase elicited by leukotriene D4 in the adherence of myelocytic leukocytes to surfaces thus is associated with an enhanced sensitivity of the Na+/H+ antiport to intracellular pH, that is, not coupled to an earlier rise in the cytosolic level of Ca+2.     

Characterization of Cronobacter recovered from dried milk and related products

Background  

Cronobacter is a recently proposed genus consisting of six genomospecies that encompass the organisms previously identified as Enterobacter sakazakii. Cronobacter are opportunistic pathogens and are known to cause serious infections in infants, particularly neonates. High case fatality rates have been associated with infections and acute sequelae can occur in survivors with severe ramifications on neurological development. Infant formula has been identified as one route of transmission for infection in infants. However, the primary reservoirs for subsequent contamination of foods with Cronobacter remain undefined due to the ubiquitous nature of these organisms. More recently, infections in adults have been reported, especially amongst the elderly and patients who are immunocompromised. To help prevent the transmission of infection, it is important to identify the main food sources for Cronobacter. The aim of this study was to identify and characterize Cronobacter isolated from dried-milk and related products available in an Egyptian food market.     

The distribution of Oligochaeta on an exposed rocky shore in southeast Ireland

The distribution of oligochaetes was studied on a wave-exposed, granite shore at Carnsore Point, County Wexford. Habitats sampled were crevices and shallow surface cracks in rock, Lichina pygmaea turf, a Mytilus edulis bed, barnacles, Laurencia pennatifida turf, Corallina officinalis turf and pools. Lichina, mussels, barnacles greater than 8 mm in height, Laurencia and Corallina were sampled quantitatively.Three species of Tubificidae and nine species of Enchytraeidae were recorded including two new species described from this locality and five which are undescribed. Oligochaetes were present in all the habitats sampled but were rare in pools and numbers were low on barnacle-covered rock except where barnacles were more than 8 mm in height. The number of species increased with decreasing tide level and was highest in lower shore crevices. Highest densities were recorded from Corallina turf and in barnacles > 8 mm. Lumbricillus semifuscus was the most widespread species, occurring in crevices and mats at all tide levels; other species had more restricted distributions. Grania species were confined to Corallina turf. Only three species of oligochaete were recorded from cracks where they were almost the only fauna present. Species diversity and density were influenced by physical structure of the habitat, particularly the amount of retained sediment.The oligochaetes are members of a rich cryptofauna in habitats which provide them with organic matter and moisture and protect them from environmental extremes and wave damage and from predators during immersion. Reasons for the scarcity of oligochaete records from exposed rocky shores and the high proportion of new species in this study are discussed.