Alterations in the cell surface proteins of Drosophila during morphogenesis

Summary Unevaginated and evaginated Drosophila imaginal discs were surface-labeled with ¹²⁵I. Relative labeling was greater in eleven peptides and lower in three peptides of evaginated discs compared to unevaginated discs. These results are compared to the effects of 20-hydroxyecdysone (20-HOE) on metabolic labeling of membrane proteins fractionated from imaginal discs, and on cell surface labeling of a hormone-responsive Drosophila tissue culture line. A group of ³⁵S-methionine labeled membrane fraction peptides whose metabolic labeling is 20-HOE dependent have isoelectric points and apparent molecular weights very similar to those of a group of proteins only labeled in iodinated evaginated discs, supporting the conclusion that these are hormone-dependent, cell surface proteins (Rickoll and Fristrom 1983). Based upon two-dimensional gel electrophoretic and immunological criteria three of the proteins showing increased labeling in evaginated discs are related to three proteins induced by 20-HOE in tissue culture cells. Two different subsets of radiolabeled peptides were observed in the imaginal discs based upon detergent solubility. Some of the proteins which are soluble in NP-40 plus urea but insoluble in NP-40 alone may be localized in the basal lamina of the imaginal discs, a structure which labels heavily with ¹²⁵I and is lacking in tissue culture cells. In discs, the majority of hormone-dependent changes in radiolabeled peptides were seen in the fraction solubilized by NP-40 and urea with a sulfhydryl reducing agent, while in tissue culture cells, the majority of differences is seen in the fraction solubilized by NP-40 only. We speculate that these proteins may be involved in similar processes, e.g., cell rearrangement, that occur during both disc morphogenesis and 20-HOE induced aggregation in tissue culture cells.This work was supported by grants CD-205 from the American Cancer Society, RR08132 from NIH to C.A.P. and GM 19937 from NIH to J.W.F.     

Calculation of the inbreeding coefficient

Wright's rule for calculating the inbreeding coefficient for an arbitrary pedigree is proven for both autosomal and X-linked loci.Supported by NSF Grant PHY-84-16691     

Cold-hardiness in several species of land snails

1. 1.|Mean supercooling points (SCP) of Anguispira alternata (Say) gradually declined in autumn and increased in spring. SCP also declined in autumn in Discus conrkhitei (Newcomb) and Gastrocopta armifera (Say).

2. 2.|Supercooling ability varied among the species, being the greatest in G. armifera.

3. 3.|Prolonged exposure of winter animals to high temperatures resulted in only a modest increase in SCP.

4. 4.|Low temperature survival of D. cronkhitei and G. armifera was slightly poorer than predicted by the mean SCP.

Natural killer (NK)-like cytotoxic activity of allospecific T cell receptor-gamma,delta+ T cell clones. Distinct receptor-ligand interactions mediate NK-like and allospecific cytotoxicity

We recently generated a series of human alloantigen-specific, CD3+,TCR-gamma,delta+ clones by stimulating CD3+,CD4-,CD8- T cells from normal individuals with allogeneic lymphoblastoid cell lines (LCL). As reported previously, these clones display cytotoxic activity against their specific stimulators but not against irrelevant LCL. Further studies of these and other TCR-gamma,delta+ clones, described in this report, indicate that most but not all of these clones express the NK cell associated marker, NKH-1 or Leu-19, and kill NK-sensitive targets such as the K562 and Molt 4 lines, but not an irrelevant LCL or NK-resistant line, Raji. TCR-gamma,delta+ clones which lacked expression of Leu-19 lysed their allospecific targets but had no detectable NK activity. The allospecific cytotoxicity of Leu-19+ and Leu-19- clones was inhibited by mAb to CD3 or the TCR delta-chain. In contrast, the NK-like activity of the Leu-19+ clones was enhanced by these antibodies over a wide range of antibody concentration. Although mAb to LFA-1 markedly inhibited both the allospecific and NK-like activity of these clones, an HLA class I framework specific mAb (W6/32) had no effect on NK-like cytolysis but did inhibit allospecific killing, suggesting that the target structures on the surface of allospecific and NK-sensitive cells are distinct. The receptors utilized by the TCR-gamma,delta+ clones to recognize NK-sensitive and allospecific targets are also distinct, since killing of NK-sensitive targets was blocked by the presence of cold (unlabeled) NK-sensitive cells but not by cold allospecific targets, whereas allospecific cytolysis was inhibited by cold allospecific targets but not by NK-sensitive cells. We conclude that some CD3+,TCR-gamma,delta+ clones exhibit NK-like as well as allospecific killing and that these two activities are mediated by distinct receptor-ligand interactions.     

Vitamin E supplementation and plasma 8-isoprostane and adiponectin in overweight subjects

Objective: Isoprostanes are a marker of oxidant stress and atherosclerotic risk, and plasma concentrations are elevated in obesity. Adiponectin is a regulator of insulin sensitivity, and low circulating levels are associated with oxidant stress and obesity. The aim of this study was to determine the effect of vitamin E supplementation on plasma concentrations of 8‐isoprostane and adiponectin in overweight/obese subjects. Research Methods and Procedures: The study was a 6‐month, randomized, double‐blind, placebo‐controlled trial in 80 overweight subjects (60 women and 20 men, BMI >27 kg/m²). Exclusion criteria were serious illness, smoking, or taking antioxidant supplements. Participants were randomized to receive 800 IU/d natural vitamin E (n = 39) or placebo (n = 41) for 3 months with an increase in the dose to 1200 IU/d for a further 3 months. Plasma 8‐isoprostane and adiponectin concentrations were measured at baseline and 3 and 6 months. Results: During 6 months of supplementation with vitamin E, plasma vitamin E concentration increased significantly (p < 0.001) by 76%, and plasma 8‐isoprostane concentrations decreased significantly (?11%, p = 0.03), whereas plasma adiponectin concentrations did not change significantly. Discussion: These findings suggest that supplementation with high‐dose vitamin E decreases systemic oxidative stress and 8‐isoprostane concentrations in overweight/obese individuals. A decrease in plasma 8‐isoprostane has the potential to reduce risk of cardiovascular disease in obesity.     

Citrobacter rodentium infection causes both mitochondrial dysfunction and intestinal epithelial barrier disruption in vivo: role of mitochondrial associated protein (Map)

Enteropathogenic Escherichia coli (EPEC) and enterohaemorrhagic E. coli are non-invasive attaching/effacing (A/E) bacterial pathogens that infect their host's intestinal epithelium, causing severe diarrhoeal disease. These bacteria utilize a type III secretion apparatus to deliver effector molecules into host cells, subverting cellular function. Mitochondrial associated protein (Map) is a multifunctional effector protein that targets host cell mitochondria and contributes to infection-induced epithelial barrier dysfunction in vitro. Unfortunately, the relevance of these actions to the pathogenesis of EPEC-induced disease is uncertain. Using Citrobacter rodentium, a mouse-adapted A/E bacterium, we found that Map colocalized with host cell mitochondria, and that in vivo infection led to a disruption of mitochondrial morphology in infected colonocytes as assessed by electron microscopy. Histochemical staining for the mitochondrial enzyme succinate dehydrogenase also revealed a significant loss of mitochondrial respiratory function in the infected intestinal epithelium; however, both pathologies were attenuated in mice infected with a Deltamap strain. C. rodentium Map was also implicated in the disruption of epithelial barrier function both in vitro and in vivo. These studies thus advance our understanding of how A/E pathogens subvert host cell functions and cause disease, demonstrating that Map contributes to the functional disruption of the intestinal epithelium during enteric infection by C. rodentium.     

Comparative sequence analysis of the region harboring the hardness locus in barley and its colinear region in rice

The ancestral shared synteny concept has been advocated as an approach to positionally clone genes from complex genomes. However, the unified grass genome model and the study of grasses as a single syntenic genome is a topic of considerable controversy. Hence, more quantitative studies of cereal colinearity at the sequence level are required. This study compared a contiguous 300-kb sequence of the barley (Hordeum vulgare) genome with the colinear region in rice (Oryza sativa). The barley sequence harbors genes involved in endosperm texture, which may be the subject of distinctive evolutionary forces and is located at the extreme telomeric end of the short arm of chromosome 5H. Comparative sequence analysis revealed the presence of five orthologous genes and a complex, postspeciation evolutionary history involving small chromosomal rearrangements, a translocation, numerous gene duplications, and extensive transposon insertion. Discrepancies in gene content and microcolinearity indicate that caution should be exercised in the use of rice as a surrogate for map-based cloning of genes from large genome cereals such as barley.     

Relative attractiveness of baits to Paratrechina longicornis (Hymenoptera: Formicidae)

Exotic ant incursions are becoming more frequent around the globe, and management with toxic baits is a suitable strategy for most species. Crazy ants, (Latreille) (Hymenoptera: Formicidae), however, are notoriously difficult to attract to commercial baits, which are generally tailored to the preferences of fire ants. We tested P. longicornis preferences for various food types and commercial ant baits. Baits trialed were commercially available products Amdro, Maxforce, Xstinguish (nontoxic monitoring version), Presto, and tuna (in spring water), sugar water (25%), boric acid (1% in 25% sugar water), and deionized water. Tuna and Xstinguish, along with sugar water and sugar water + boric acid, were the most attractive baits to P. longicornis foragers. The granular baits (Maxforce, Amdro, and Presto) were not as attractive to P. longicornis foragers. A decrease in temperature from summer (30 degrees C) to autumn (23 degrees C) trials did not seem to affect the food preferences of P. longicornis. Although P. longicornis recruitment was substantially lower during trials where there was concurrent high native ant abundance and diversity, P. longicornis still recruited to preferred baits in numbers higher than any other species. Given that tuna is impractical for management programs, the effectiveness of boric acid, sweet liquid baits in eliminating P. longicornis colonies should be compared with that of the toxic version of Xstinguish. If both are effective at eliminating colonies, we recommend sweet liquid baits containing boric acid be used for small-scale incursions (one or two nests), but a more practicable solid bait, such as Xstinguish, be used for larger scale incursions (numerous nests).