Lipoxygenases and atherosclerosis: protection versus pathogenesis

15 lipoxygenase (15LO) is a lipid-oxidizing enzyme that is considered to contribute to the formation of oxidized lipids in atherosclerotic lesions. Monocyte-macrophages are the key cells that express 15LO in atherosclerotic lesions. In this review, we examine the evidence for 15LO involvement in atherogenesis and explore and evaluate the potential mechanisms whereby 15LO may contribute to the oxidation of LDL by monocyte-macrophages. We also describe several possible pro- versus anti-atherogenic functions that may be mediated by various products of 15LO lipid oxidation. Central pathways involved in regulating 15LO expression and activity that may serve as future targets for intervention and regulation of this enzyme are also reviewed.     

Monocyte 15-lipoxygenase expression is regulated by a novel cytosolic signaling complex with protein kinase C delta and tyrosine-phosphorylated Stat3

Our previous studies demonstrated that the IL-13-induced 15-lipoxygenase expression in primary human monocytes is regulated by the activation of both Stat1 and Stat3 and by protein kinase C (PKC)delta. IL-13 stimulated the phosphorylation of Stat3 on both Tyr705 and Ser727. In this study we show that IL-13 induces the association of PKCdelta with Stat3, not with Stat1, and is required for Stat3 Ser727 phosphorylation. We found a novel IL-13-dependent cytosolic signaling complex of PKCdelta and tyrosine-phosphorylated Stat3. A tyrosine kinase inhibitor blocked PKCdelta association with Stat3 as well as Stat3 Ser727 phosphorylation. We therefore hypothesized that tyrosine phosphorylation was required for Stat3 interaction with PKCdelta and subsequent PKCdelta-dependent phosphorylation of Stat3 Ser727. We developed an efficient transfection protocol for human monocytes. Expression of Stat3 containing a mutation in Tyr705 inhibited the association of PKCdelta with Stat3 and blocked Stat3 Ser727 phosphorylation, whereas transfection with wild-type Stat3 did not. Furthermore, by transfecting monocytes with Stat3 containing mutations in Tyr705 or Ser727 or with wild-type Stat3, we demonstrated that both Stat3 tyrosine and serine phosphorylations are required for optimal binding of Stat3 with DNA and maximal expression of 15-lipoxygenase, an important regulator of inflammation and apoptosis.     

Membrane lipids and ethanol tolerance in the mouse. The influence of dietary fatty acid composition

Mice of the TO Swiss strain received diets containing different amounts of saturated or unsaturated fat throughout life. These diets produced characteristic changes in cardiac phospholipid fatty acid composition, but produced no significant differences in fatty acid composition of phospholipids from a crude membrane fraction of brain. When littermates of these animals were exposed to ethanol vapour in an inhalation chamber it was observed that mice which had received a diet high in saturated fat lost the righting reflex at an estimated concentration of ethanol in blood higher than that required for mice receiving a control diet, or a diet rich in polyunsaturated fat. Analysis of the brain membrane fraction from those animals which had received ethanol revealed that mice receiving the highly saturated fat diet now had a significantly greater proportion of saturated fatty acids in brain membrane phospholipids. These results are discussed in relation to the hypothesis that brain membrane lipid composition may influence the behavioural response to ethanol.     

Phylogenetic relationships among prokaryotic and eukaryotic catalases

Seventy-four catalase protein sequences, including 29 bacterial, 8 fungal, 7 animal, and 30 plant sequences, were compiled, and 70 were used for phylogenetic reconstruction. The core of the resulting tree revealed unique, separate groups of plant and animal catalases, two groups of fungal catalases, and three groups of bacterial catalases. The only overlap of kingdoms occurred within one branch and involved fungal and bacterial large-subunit enzymes. The other fungal branch was closely linked to the group of animal enzymes. Group I bacterial catalases were more closely related to the plant enzymes and contained such diverse taxa as the Gram-positive Listeria seeligeri, Deinocococcus radiodurans, and gamma-proteobacteria. Group III bacterial sequences were more closely related to fungal and animal sequences and included enzymes from a broad range of bacteria including high- and low-GC Gram positives, proteobacteria, and a bacteroides species. Group II was composed of large-subunit catalases from diverse sources including Gram positives (low-GC Bacilli and high-GC Mycobacteria), proteobacteria, and species of the filamentous fungus Aspergillus. These data can be interpreted in terms of two gene duplication events that produced a minimum of three catalase gene family members that subsequently evolved in response to environmental demands. Horizontal gene transfer may have been responsible for the group II mixture of bacterial and fungal large-subunit catalases.      

Mutations in Escherichia coli altering an apurinic endonuclease, endonuclease II, and exonuclease III and their effect on in vivo sensitivity to methylmethanesulfonate.

The levels of endonuclease II, an apurinic endonuclease, and exonuclease III in the parent strains (AB 1157) of Escherichia coli and in various mutants were determined by chromatography on DEAE-cellulose. AB 3027 and NH 5016 lacked endonuclease II and exonuclease III. BW 2001 lacked the apurinic endonuclease and exonuclease III while BW 2007, BW 9093, and BW 9059 lacked only exonuclease III. Deletion mutants BW 9101 and BW 9109 lacked all three enzymes. The latter mutants locate the genes for the two endonucleases in the region of exonuclease III (chith) of 38.2 min (White et al., 1976). All of the mutants which were sensitive to methylmethanesulfonate in vivo lacked exonuclease III, but not all mutants lacking exonuclease III were MMS sensitive. The deletion mutants and NH 5016 were the exceptions.     

In vitro knockout of human p47phox blocks superoxide anion production and LDL oxidation by activated human monocytes

We previously reported that superoxide dismutase (SOD) blocked human monocyte oxidation of LDL and therefore concluded that superoxide anion (O(2)(.-)) was required for oxidation. Others, however, have suggested that SOD may inhibit by mechanisms alternative to the dismutation of O(2)(.-). This study definitively addresses the involvement of O(2)(.-) in monocyte oxidation of LDL. Using an antisense ODN designed to target p47phox mRNA, we found that treatment of monocytes with antisense ODN caused a substantial and selective decrease in expression of p47phox protein, whereas sense ODN was without effect. Corresponding functional assays demonstrated that antisense ODN inhibited production of O(2)(.-). As sense ODN caused no inhibition of O(2)(.-) production, these results suggested that inhibition of p47phox expression caused reduction in O(2)(.-) production. Evaluation of the contribution of O(2)(.-) production to monocyte-mediated oxidation of LDL lipids confirmed that O(2)(.-) production is required for LDL lipid oxidation as antisense ODN treatment significantly inhibited LDL oxidation whereas sense ODN treatment caused no inhibition. This is the first report of the reduction of NADPH oxidase activity in intact human monocytes by directly targeting the mRNA of a significant member of this enzyme complex. Our results provide convincing data that O(2)(.-) is indeed required for monocyte-mediated LDL oxidation.     

Vice-anvil use in nut processing by two Corvus species

Woodpeckers and certain passerine species secure encased food in the environment in various ways to facilitate the extraction of the contents with their bills. They do this by securing the food items in locations such as crevices and holes, newly defined in this paper as ‘vice-anvils’. Here I report that free-living New Caledonian crows (Corvus moneduloides) and rooks (Corvus frugilegus, in New Zealand) also use vice-anvils to process candlenuts and walnuts, respectively. New Caledonian crows placed candlenut sections in vice-anvils to aid kernel extraction, after the candlenuts had been dropped onto an anvil to break them open. In contrast, rooks used vice-anvils to secure walnuts while they broke the shell with their bills. Long-term use by rooks of a vice-anvil in a tree had produced a ‘purpose-made’ nut-cracking site. My findings extend the persistent use of specific vice-anvils to Corvus species and further demonstrate their innovative and flexible foraging behaviour.     

ACUTE EFFECTS OF MOVEMENT VELOCITY ON BLOOD LACTATE AND GROWTH HORMONE RESPONSES AFTER ECCENTRIC BENCH PRESS EXERCISE IN RESISTANCE-TRAINED MEN

This study aimed to compare the effects of different velocities of eccentric muscle actions on acute blood lactate and serum growth hormone (GH) concentrations following free weight bench press exercises performed by resistance-trained men. Sixteen healthy men were divided into two groups: slow eccentric velocity (SEV; n = 8) and fast eccentric velocity (FEV; n = 8). Both groups performed four sets of eight eccentric repetitions at an intensity of 70% of their one repetition maximum eccentric (1RMecc) test, with 2-minute rest intervals between sets. The eccentric velocity was controlled to 3 seconds per range of motion for SEV and 0.5 seconds for the FEV group. There was a significant difference (P < 0.001) in the kinetics of blood lactate removal (at 3, 6, 9, 15, and 20 min) and higher mean values for peak blood lactate (P = 0.001) for the SEV group (9.1 ± 0.5 mM) compared to the FEV group (6.1 ± 0.4 mM). Additionally, serum GH concentrations were significantly higher (P < 0.001) at 15 minutes after bench press exercise in the SEV group (1.7 ± 0.6 ng · mL⁻¹) relative to the FEV group (0.1 ± 0.0 ng · mL⁻¹). In conclusion, the velocity of eccentric muscle action influences acute responses following bench press exercises performed by resistance-trained men using a slow velocity resulting in a greater metabolic stress and hormone response.     

Field swimming performance of bluegill sunfish,Lepomis macrochirus: implications for field activity cost estimates and laboratory measures of swimming performance

Mobility is essential to the fitness of many animals, and the costs of locomotion can dominate daily energy budgets. Locomotor costs are determined by the physiological demands of sustaining mechanical performance, yet performance is poorly understood for most animals in the field, particularly aquatic organisms. We have used 3‐D underwater videography to quantify the swimming trajectories and propulsive modes of bluegills sunfish (Lepomis macrochirus, Rafinesque) in the field with high spatial (1–3 mm per pixel) and temporal (60 Hz frame rate) resolution. Although field swimming trajectories were variable and nonlinear in comparison to quasi steady‐state swimming in recirculating flumes, they were much less unsteady than the volitional swimming behaviors that underlie existing predictive models of field swimming cost. Performance analyses suggested that speed and path curvature data could be used to derive reasonable estimates of locomotor cost that fit within measured capacities for sustainable activity. The distinct differences between field swimming behavior and performance measures obtained under steady‐state laboratory conditions suggest that field observations are essential for informing approaches to quantifying locomotor performance in the laboratory.