Increased 12-HETE production in bovine lymphocytes during selenium deficiency

When peripheral blood lymphocytes were incubated with arachidonic acid in the presence of Ca++ ionophore (A23187), the cells from the selenium-deficient dairy cows produced significantly greater quantities of 12-hydroxyeicosatetraenoic acid (12-HETE) than the cells from the selenium-supplemented animals. The major product of reaction was verified as 12-HETE by cochromatography with a 12-HETE standard on an HPLC and structural analysis by GC-MS. Additionally, concanavalin A-stimulated lymphocyte proliferation was significantly decreased in cells from the Se-deficient cows. Furthermore, 12-HETE generated by the A23187-stimulated lymphocytes inhibited lymphocyte proliferation when added to Se-supplemented cell cultures. These observations suggest that self-regulation of lymphocyte proliferation might be mediated by 12-HETE production, especially during an altered nutritional state such as Se deficiency.     

Leukocyte adhesion molecule expression during intense resistance exercise

We hypothesized that expression of L-selectinand very late antigen-4 (VLA-4) integrin adhesionmolecules would influence cell type-specific redistribution duringexercise. Women subjects performed six sets of 10-repetition maximumsquats. L-selectin and VLA-4 integrin were measured by using flowcytometry pre- and postexercise on peripheral blood neutrophils andlymphocytes (n = 29 subjects) andlymphocyte subsets (n = 70 subjects),respectively. Neutrophil concentration increased 41.8%(P < 0.001), whereas the percentexpressing L-selectin was unchanged (79%). Lymphocyte concentrationincreased 61.8% (P < 0.001). Thepercent of T cells expressing L-selectin decreased from 73.5 ± 8.9 to 68.2 ± 11.4% (P < 0.001); the combined population of natural killer and B cells expressing L-selectin decreased from 80.4 ± 22.5 to 62.7 ± 25.8% (P < 0.001).VLA-4 integrin was expressed by nearly all lymphocytes both pre- andpostexercise. The proportional decrease in L-selectin positive cellscould have resulted from 1) sheddingof L-selectin, 2) selective entry ofL-selectin-negative subsets, or 3)selective removal of L-selectin-positive subsets.

     

The interaction of [3H]TPA with bovine lymph node lymphocytes in vitro

The potent tumor promoter, 12-O-tetradecanoyl-phorbol-13-acetate (TPA), is an effective modulator of DNA synthesis in bovine lymph node lymphocytes in culture. The effect of TPA on cellular proliferation depends to a great extent on the length of exposure and the mitogenic stimulus. Addition of TPA along with mitogenic lectins enhances DNA synthesis. Exposure to TPA for 2 days before addition of lectins depresses DNA synthesis. There is evidence that some inhbitory material other than TPA is formed during the longer incubation. Therefore, in this study, we used [³H]TPA to determine (i) the amount of material retained by the cells after preincubation and (ii) if TPA was metabolized during this culture period. We found that after incubation with 10^?7 M, [³H]TPA, less than 3% of the radioactivity was retained by the cells. This was released by incubation in fresh medium. All of the cell associated material appeared to be TPA. However, the TPA in the medium was degraded by about 30% during a 2-day incubation to 12-O-tetradecanoyl-phorbol (TP), phorbol-13-acetate (PA) and phorbol. The source of the hydrolytic activity appeared to be serum because the same effect was seen in the absence of cells but was not seen in the absence of serum. These metabolites when added directly to the lymphocytes had no effect on DNA synthesis. Moreover, the amount of TPA retained by the cells and released into the medium was too small to account for the inhibitory activity of medium from TPA-treated cells. Studies are in progress to determine the nature of the inhibitory material after exposure to TPA.     

Visualization of protein kinases in lymphocytes stimulated to proliferate with concanavalin A or inhibited with a phorbol ester

Stimulation of lymphocytes with a mitogenic lectin such as concanavalin A (ConA) results in differentiation and cell division. Among the changes which occur after stimulation are increases in phosphorylation of proteins and in protein kinase activity. We used a high-resolution, nondenaturing gel system to separate and visualize protein kinases in situ. We have clearly identified both autophosphorylating and substrate-dependent kinases. One band of cyclic AMP-dependent kinase activity was significantly enhanced in lectin-stimulated cells. In contrast, treatment of the cells with phorbol ester under conditions which depress stimulation caused a decrease in the activity of one kinase.     

A unified local objective function for optimally selecting SNPs on arrays for agricultural genomics applications

Over the years, ad-hoc procedures were used for designing SNP arrays, but the procedures and strategies varied considerably case by case. Recently, a multiple-objective, local optimization (MOLO) algorithm was proposed to select SNPs for SNP arrays, which maximizes the adjusted SNP information (E score) under multiple constraints, e.g. on MAF, uniformness of SNP locations (U score), the inclusion of obligatory SNPs and the number and size of gaps. In the MOLO, each chromosome is split into equally spaced segments and local optima are selected as the SNPs having the highest adjusted E score within each segment, conditional on the presence of obligatory SNPs. The computation of the adjusted E score, however, is empirical, and it does not scale well between the uniformness of SNP locations and SNP informativeness. In addition, the MOLO objective function does not accommodate the selection of uniformly distributed SNPs. In the present study, we proposed a unified local function for optimally selecting SNPs, as an amendment to the MOLO algorithm. This new local function takes scalable weights between the uniformness and informativeness of SNPs, which allows the selection of SNPs under varied scenarios. The results showed that the weighting between the U and the E scores led to a higher imputation concordance rate than the U score or E score alone. The results from the evaluation of six commercial bovine SNP chips further confirmed this conclusion.     

Calcium-dependent activation of lymphocytes by ionophore, A23187, and a phorbol ester tumor promoter

The phorbol ester tumor promoter 12-O-tetradecanoylphorbol-13-acetate (TPA) and the calcium ionophore, A23187, have similar effects on many different cells. For example, both show mitogenic and comitogenic activities for lymphocytes. It had been suggested that some of TPA's effects are due to its ability to act as a calcium ionophore. In order to test this idea, we compared the ability of TPA and ionophore to synergize with concanavalin A (Con A) in a two-phase system of lymphocyte mitogenesis. We found that ionophore was most comitogenic with Con A when present in the early phase of stimulation. TPA was only comitogenic when present in the late phase. Ionophore and TPA could not replace one another in the system. However, both ionophore and TPA together could replace Con A and stimulate DNA synthesis when they were presented to the cells in the sequential order of ionophore followed by TPA. Both compounds required the presence of external calcium to be effective.     

Prevention of the TPA-mediated down-regulation of protein kinase C

Protein kinase C activity in lymphocytes was down-regulated upon exposure of the cells to 12-0-tetradecanoylphorbol-13 acetate. This down-regulation was prevented by preincubating the cells with sphingosine, a reported protein kinase C inhibitor. Two other protein kinase C inhibitors, palmitoylcarnitine and phloretin, were ineffective in preventing down-regulation by the phorbol ester.     

Novel visual-cue-based sticky traps for monitoring of emerald ash borers, Agrilus planipennis (Col., Buprestidae)

We examined various methods of trapping emerald ash borers (EAB), Agrilus planipennis Fairmaire, using solely visual cues based on previous work that has documented the importance of visual cues in EAB mate location. Here, we give the results of two of these methods, coloured sticky cards (yellow or blue), or live ash leaves covered with spray-on adhesive to which dead male EAB visual lures had been pinned. Feral male beetles were captured effectively on the sticky traps made from dead male EAB on ash leaves. These sticky-leaf-traps captured more male EAB when deployed in high-population density areas than low-density areas, but did capture EAB even at lower population densities. More feral males were captured on these traps when they were placed higher in the trees, regardless of the population density of EAB. Very few feral female EAB were captured using the sticky-leaf-traps. This novel method of EAB trapping may allow 'real-time' population detection and monitoring of EAB adults during the active flight period rather than locating larval galleries during the autumn and winter after adult flight and attack. Feral male beetles were also captured using standard yellow- or blue-coloured sticky cards to which male EAB had been affixed with adhesive; however, this type of trap was much less effective overall than using the sticky-leaf-traps. Furthermore, Agrilus cyanescens , a species similar in colour to EAB but smaller in size, showed a strong response to blue-coloured sticky traps to which dead male EAB had been affixed with adhesive, suggesting a general use of visual cues in the mating systems of some of the other Buprestidae as well.