Effect of Leaf Wetness Duration, Temperature, and Conidial Inoculum Dose on Apple Scab Infections.

Apple seedlings (2 months old, 'Idared' × 'Golden Delicious') were inoculated with conidia of Venturia inaequalis in order to study the effects of inoculum dose and leaf wetness duration on development of apple scab symptoms. For each experiment, the C3 curve (indicating heavy infection levels) was used as the basis for relating infection to temperature and leaf wetness duration. In one series of experiments, seedlings were treated with inoculum doses of 1.5, 5.4, 15.6, 32.2, 81.2, and 250 × 103 conidia/ml and leaves were kept wet during C3 infection periods at temperatures of 6, 11, 16, and 22°C. At all four temperatures, disease incidence (scab lesions/plant) increased with increasing inoculum doses up to about 81.2 × 103 conidia/ml. Disease incidence was lower at 22°C than at the other temperatures. In a second series of experiments, seedlings inoculated with 10 × 103 conidia/ml were kept moist for infection periods ranging from 0.6 to 2.0 times the C3 leaf wetness duration curve at 6, 11, 16, and 22°C. Disease incidence increased with increasing duration of leaf wetness and generally leveled off between 150 and 200% of the C3 curve. At this inoculum dose (10 × 103 conidia/ml), doubling the leaf wetness duration indicated by the C3 curve resulted in high disease incidence, similar to levels obtained with a higher inoculum (250 × 103 conidia/ml) and shorter wetness period (1.0 C3).

Selective inhibition of platelet macroaggregate formation by a recombinant heparin-binding domain of human thrombospondin.

Thrombospondin (TSP) is a platelet alpha-granule adhesive protein that plays a critical role in the stabilization of thrombus by promoting the formation of platelet macroaggregates. We have recently shown that a monoclonal antibody (mAb) to the NH2-terminal heparin-binding domain of TSP, MAII, inhibits platelet aggregation induced by thrombin in a dose-dependent manner. In this study, we have expressed in Escherichia coli two recombinant proteins comprising residues 1 to 174 (TSP18) and 1 to 242 (TSP28) of TSP. After purification, both proteins reacted equally well with mAb MAII, whereas the reactivity of TSP18 for heparin was lower than that of TSP28 or native TSP. At micromolar concentrations, TSP18 and TSP28 inhibited the second wave of platelet aggregation and the concomitant release of [14C]5-hydroxytryptamine induced by ADP in citrated platelet-rich plasma as well as aggregation and secretion induced by a low concentration of thrombin in washed platelet suspensions. The proteins did not inhibit surface expression of endogenous TSP on activated platelets, as measured by the binding of radiolabeled mAb 5G11, indicating that they did not interfere with the primary binding of TSP to the plasma membrane. In contrast, in a solid-phase binding assay, the proteins inhibited in a dose-dependent manner (IC50, 0.1 and 0.06 mumol/L for TSP18 and TSP28, respectively) the binding of radiolabeled TSP to surface-adsorbed fibrinogen. Furthermore, specific and saturable binding of the proteins to immobilized fibrinogen was demonstrated by enzyme-linked immunosorbent assay. The results suggest that interaction between the heparin-binding domain of TSP and membrane-bound fibrinogen may be critical in the platelet aggregation/secretion process.

At high dietary levels ethanol alters the structure of mid- and hindgut epithelial cells of Drosophila melanogaster larvae.

The midgut of Drosophila melanogaster is a site of alcohol dehydrogenase (ADH) activity, the enzyme that catalyzes the first step in the major pathway for ethanol degradation. The effects of different levels of dietary ethanol on the ultrastructures of the guts of larvae of the Canton-S wild-type strain and the ADH-deficient, Adhn2, strain were ascertained. In wild-type larvae fed an ethanol-free, defined medium, the foregut epithelium was characterized by few glycogen rosettes and sparse microvilli that protruded into the gut's thick lumen lining. The midgut epithelium was typical of cells involved in absorption and active transport with abundant microvilli on the apical surface and membrane infoldings on the basal surface. In place of microvilli, the apical surface of the hindgut had membrane infoldings. The apical surfaces of both the mid- and hindgut epithelium were covered by a thick, electron-dense peritrophic membrane consisting of chitin. In both strains the subcellular damage that was correlated with ethanol levels in the diet was confined to the midgut and hindgut regions. Damage to gut cells in the form of disrupted mitochondria, dilated rough endoplasmic reticulum, low densities of glycogen rosettes and protein granules, high numbers of autophagic vacuoles, and the presence of myelin whirls was extensive in Canton-S strain larvae fed a high ethanol diet. A low dietary concentration of ethanol induced changes in gut ultrastructure of Adhn2 larvae similar to the changes that were observed in wild-type larvae fed the higher ethanol concentrations, but the basal infoldings were more dilated in the Adhn2 larvae. At high dietary concentrations the disruption of mid- and hindgut cells by ethanol appeared great enough to interfere with the digestion and absorption of nutrients.