Expression and functional characterization of the cancer-related serine protease, human tissue kallikrein 14

Human tissue kallikrein 14 (KLK14) is a novel extracellular serine protease. Clinical data link KLK14 expression to several diseases, primarily cancer; however, little is known of its (patho)-physiological role. To functionally characterize KLK14, we expressed and purified recombinant KLK14 in mature and proenzyme forms and determined its expression pattern, specificity, regulation, and in vitro substrates. By using our novel immunoassay, the normal and/or diseased skin, breast, prostate, and ovary contained the highest concentration of KLK14. Serum KLK14 levels were significantly elevated in prostate cancer patients compared with healthy males. KLK14 displayed trypsin-like specificity with high selectivity for P1-Arg over Lys. KLK14 activity could be regulated as follows: 1) by autolytic cleavage leading to enzymatic inactivation; 2) by the inhibitory serpins alpha1-antitrypsin, alpha2-antiplasmin, antithrombin III, and alpha1-antichymotrypsin with second order rate constants (k(+2)/Ki) of 49.8, 23.8, 1.48, and 0.224 microM(-1) min(-1), respectively, as well as plasminogen activator inhibitor-1; and 3) by citrate and zinc ions, which exerted stimulatory and inhibitory effects on KLK14 activity, respectively. We also expanded the in vitro target repertoire of KLK14 to include collagens I-IV, fibronectin, laminin, kininogen, fibrinogen, plasminogen, vitronectin, and insulin-like growth factor-binding proteins 2 and 3. Our results indicate that KLK14 may be implicated in several facets of tumor progression, including growth, invasion, and angiogenesis, as well as in arthritic disease via deterioration of cartilage. These findings may have clinical implications for the management of cancer and other disorders in which KLK14 activity is elevated.     

Tumor-infiltrating dendritic cell precursors recruited by a beta-defensin contribute to vasculogenesis under the influence of Vegf-A

The involvement of immune mechanisms in tumor angiogenesis is unclear. Here we describe a new mechanism of tumor vasculogenesis mediated by dendritic cell (DC) precursors through the cooperation of beta-defensins and vascular endothelial growth factor-A (Vegf-A). Expression of mouse beta-defensin-29 recruited DC precursors to tumors and enhanced tumor vascularization and growth in the presence of increased Vegf-A expression. A new leukocyte population expressing DC and endothelial markers was uncovered in mouse and human ovarian carcinomas coexpressing Vegf-A and beta-defensins. Tumor-infiltrating DCs migrated to tumor vessels and independently assembled neovasculature in vivo. Bone marrow-derived DCs underwent endothelial-like differentiation ex vivo, migrated to blood vessels and promoted the growth of tumors expressing high levels of Vegf-A. We show that beta-defensins and Vegf-A cooperate to promote tumor vasculogenesis by carrying out distinct tasks: beta-defensins chemoattract DC precursors through CCR6, whereas Vegf-A primarily induces their endothelial-like specialization and migration to vessels, which is mediated by Vegf receptor-2.     

Prey‐mediated changes in the selectivity of the predator Macrolophus pygmaeus (Heteroptera: Miridae)

When foraging in communities with mixed prey, generalist predators may be confronted with prey species that differ in quality, size and mobility and interact with one another. To examine prey selection, predation by Macrolophus pygmaeus (Heteroptera: Miridae) was recorded by providing a diet of either one or two prey species of Myzus persicae (third‐instar nymphs), Aphis gossypii (fourth‐instar nymphs), Trialeurodes vaporariorum (third‐instar nymphs) and Ephestia kuehniella (eggs). In the experiments, prey mobility, prey quality and prey biomass were considered. The biomass consumed by the predator was dependent on the combination of prey species and the quantity of biomass offered. In choice experiments with diets mixed of two prey species at equal densities, the predation to A. gossypii was significantly reduced in the presence of E. kuehniella but the rate of consumption of M. persicae, T. vaporariorum and E.kuehniella was not significantly affected by the coexistence of any other species in the mixed prey diet. When equal amounts of biomass from two prey species were provided in combination, the total consumed biomass was significantly reduced in the mixed prey diets composed of E. kuehniella eggs and aphid nymphs. Thus, under the mixed‐prey situation, prey selection by predators may be affected by interactions among prey species differing in traits such as quality, mobility and size.     

A method for laboratory studies on the polyphagous predator Macrolophus pygmaeus (Hemiptera: Miridae)

A method for studying the life history parameters of Macrolophus pygmaeus Rambur was developed and described. Fecundity of the predator was studied by placing one female and one male in a plastic cylinder in which a stem piece was used as an ovipositional substrate. Smaller cylinders were used to study the percentage of egg hatch of M. pygmaeus. Results demonstrated that this method is suitable for studying the fecundity and egg eclosion of M. pygmaeus. Allowing for easy modification, the method offers suitable conditions for maintenance of both insect and plant material and can be inexpensively modified for different experiments and the parts are easily constructed. The proposed method could also be employed for similar studies on other predatory hemipterans such as mirids and anthocorids and very likely on other phytophagous insect species that lay their eggs in or on plant stems.     

Are multiple predator effects directed by prey availability?

Antagonistic/synergistic interactions among predators foraging on the same prey have been assumed to play a major role in shaping community structure. Studies in systems with multiple predator species have shown that the strength of these interactions may not be predictable and is largely dependent on individual behavioural traits, species density and habitat complexity. Although the association of prey consumption and satiation of a foraging predator has long been recognized, there has been relatively little research on how prey availability affects multiple predators’ effects. In this work, we present a framework to investigate the variation in two coexisting/competing predators’ effects on prey risk as affected by the prey availability rate. Functional responses by each predator species were first studied in single-predator treatments. Then, the intra- and inter-specific competition was investigated by employing additive and substitutative experimental designs to highlight the nature of multiple effects. Intra- and interspecific interactions were found to be similar and there was risk reduction, and risk enhancement for the prey at intermediate and high levels, respectively, according to the multiplicative risk model (MRM). The results indicated that when similar predators are concerned, the outcomes of MRM may vary according to the functional response curve of these predators. Thus, studies involving a wide range of prey densities are required to explore the nature of interactions. Moreover, this kind of experimental data can contribute to unravelling complexities in theoretical approaches by earlier studies and ultimately promote understanding the effect of multiple predators on prey population regulation.     

The influence of temperature, photoperiod and plant type on the predation rate of Macrolophus pygmaeus on Myzus persicae

The effect of temperature and photoperiod on the rate of predation of nymphs and adults of the predator Macrolophus pygmaeus was studied using Myzus persicae prey feeding on egg-plant and pepper plants. The experiments were conducted at three photoperiods (16L:8D, 12L:12D and 8L:16D), three temperatures (20, 25 and 30 °C), and at 65% r.h. The rate of predation increased with temperature. Predation rate was affected by photoperiod on pepper but not on egg-plant. Females and fifth instar nymphs were the most voracious stages followed by third and fourth instar nymphs and males. First and second instar nymphs consumed far fewer aphids. Predation rate was higher on leaves of pepper than egg-plant, especially at 30 °C. Variation in the efficacy of this predator is discussed.