Essential role for the p110delta phosphoinositide 3-kinase in the allergic response.

Inflammatory substances released by mast cells induce and maintain the allergic response. Mast cell differentiation and activation are regulated, respectively, by stem cell factor (SCF; also known as Kit ligand) and by allergen in complex with allergen-specific immunoglobulin E (IgE). Activated SCF receptors and high-affinity receptors for IgE (FcvarepsilonRI) engage phosphoinositide 3-kinases (PI(3)Ks) to generate intracellular lipid second messenger signals. Here, we report that genetic or pharmacological inactivation of the p110delta isoform of PI(3)K in mast cells leads to defective SCF-mediated in vitro proliferation, adhesion and migration, and to impaired allergen-IgE-induced degranulation and cytokine release. Inactivation of p110delta protects mice against anaphylactic allergic responses. These results identify p110delta as a new target for therapeutic intervention in allergy and mast-cell-related pathologies.

Glucose-potentiated chemotaxis in human vascular smooth muscle is dependent on cross-talk between the PI3K and MAPK signaling pathways.

Atheroma formation involves the movement of vascular smooth muscle cells (VSMC) into the subendothelial space. The aim of this study was to determine the involvement of PI3K and MAPK pathways and the importance of cross-talk between these pathways, in glucose-potentiated VSMC chemotaxis to serum factors. VSMC chemotaxis occurred in a serum gradient in 25 mmol/L glucose (but not in 5 mmol/L glucose) in association with increased phosphorylation (activation) of Akt and ERK1/2 in PI3K and MAPK pathways, respectively. Inhibitors of these pathways blocked chemotaxis, as did an mTOR inhibitor. VSMC expressed all class IA PI3K isoforms, but microinjection experiments demonstrated that only the p110beta isoform was involved in chemotaxis. ERK1/2 phosphorylation was reduced not only by MAPK pathway inhibitors but also by PI3K and mTOR inhibitors; when PI3K was inhibited, ERK phosphorylation could be induced by microinjected activated Akt, indicating important cross-talk between the PI3K and ERK1/2 pathways. Glucose-potentiated phosphorylation of molecules in the p38 and JNK MAPK pathways inhibited these pathways but did not affect chemotaxis. The statin, mevinolin, blocked chemotaxis through its effects on the MAPK pathway. Mevinolin-inhibited chemotaxis was restored by farnesylpyrophosphate but not by geranylgeranylpyrophosphate; in the absence of mevinolin, inhibition of farnesyltransferase reduced ERK phosphorylation and blocked chemotaxis, indicating a role for the Ras family of GTPases (MAPK pathway) under these conditions. In conclusion, glucose sensitizes VSMC to serum, inducing chemotaxis via pathways involving p110beta-PI3K, Akt, mTOR, and ERK1/2 MAPK. Cross-talk between the PI3K and MAPK pathways is necessary for VSMC chemotaxis under these conditions.

Regulation of breast cancer cell chemotaxis by the phosphoinositide 3-kinase p110delta.

Class IA phosphoinositide 3'-kinases (PI3Ks) regulate many cellular processes downstream of tyrosine kinases and Ras. Despite a clear implication of PI3K in cancer, little is known about the distribution of the different PI3K isoforms in malignant cells. We screened a large panel of tissues and cell lines for expression of class IA PI3Ks, and document a ubiquitous expression of the p110alpha and p110beta isoforms but a variable and more restricted tissue distribution of the p110delta isoform. Originally found in WBCs, p110delta was also detected in some nonhematopoietic cell types especially those of breast or melanocytic origin, both in the untransformed and transformed state. Isoform-specific neutralization of PI3K isoforms in breast cancer cell lines (by PI3K antibody microinjection or a p110delta-selective pharmacological inhibitor) demonstrated that p110delta is the most important class IA PI3K in the regulation of epidermal growth factor-driven motility in vitro, controlling the directionality and, to a lesser extent, the speed of migration. In contrast, p110beta was required for the direction but not the speed of migration, whereas p110alpha did not impact on either of these parameters. These results show a nonredundant function of PI3K isoforms downstream of the epidermal growth factor receptor and indicate that the presence of p110delta may confer breast cancer cells with selective migratory capacities. The potential clinical implications of p110delta expression in non-WBC-derived tumors are discussed.

Post-Processing Temperature Rise in Foods: Conventional Hot Air and Microwave Ovens 1.

Measurement of temperature rise (N = 5 replications) in water (1000 ml), chicken frankfurters (46 ± 2 g/frankfurter) and cake cones (40 g/cone) after conventional hot air (160°C) and after microwave (2450 MHz; 50% and 100% power of 645 ± 25 W) processing indicated that temperature rise occurred more often in products heat-processed in microwave than in hot air ovens. Duration and extent of post-processing temperature rise (PPTR) in beef loaf patties (150 g/patty), pork and turkey roasts (approximately 2.3 kg/roast) and turkey casserole (0.9 kg/casserole) prepared in microwave ovens was quantified during three replications. Although present, PPTR should not change temperature objectives for domestic microwave processing of foods because of the extensive variability of duration and extent in PPTR within and among experimental products tested. However, PPTR should be given consideration when commercial products to be processed in microwave ovens and those used in mathematical modeling of microwave cooking/heating procedures are designed.

Effectiveness of Cold-serving Units Using Two Cold-holding Methods in Foodservice Operations 1.

Effectiveness of two cold-holding methods commonly used to maintain temperatures of products held on cold-serving units (CSU) was determined by time-temperature and bacterial growth patterns of three products. Products used were bulk (2.27 kg) and portioned (100 g) cottage cheese, portioned (100 g) tuna salad, and deviled eggs halves (100 ± 10 g). All products were held on a cold-serving unit using the mechanical/ice cold-holding method (mechanical cooling used in combination with 3 to 10 cm ice) for 24 h (control; laboratory setting), as well as on three separate cold-serving units using the mechanical cold-holding method (at three university residence hall field sites under actual operating conditions) for 4 h (maximum length of service). Temperatures of all bulk and portioned products held on CSUs using the mechanical/ice cold-holding method (initial temperatures of food were 4 to 8.2°C) were >7.2°C after 2 h with a 50% load factor. When the mechanical cold-holding method was used, all portioned products (initial temperatures were 8.2 to 11°C) were <7.2°C after 2 h with a 75% load factor. Temperature differences between the mechanical and mechanical/ice cold-holding methods were attributed to ice on the cold-serving unit. The ice insulated the products from the mechanically cooled basin and allowed internal temperatures of the products to increase. Statistical significance for bacterial growth patterns was reported only for products held on cold-serving units using the mechanical/ice cold-holding method: mesophilic growth in deviled eggs (p<0.05) and psychrotrophic growth in tuna salad (p<0.001). As expected, bulk cottage cheese had a significantly higher temperature over time (p<0.05) than did portioned cottage cheese for both methods of cold-holding. Based on results of this study, portioned foods on cold-serving units should be held less than 2 h when the mechanical/ice cold-holding method is used, or up to 4 h when the mechanical cold-holding method is available.

Class I phosphoinositide 3-kinase p110beta is required for apoptotic cell and Fcgamma receptor-mediated phagocytosis by macrophages.

Phosphoinositide 3-kinases (PI3Ks) play an important role in a variety of cellular functions, including phagocytosis. PI3Ks are activated during phagocytosis induced by several receptors and have been shown to be required for phagocytosis through the use of inhibitors such as wortmannin and LY294002. Mammalian cells have multiple isoforms of PI3K, and the role of the individual isoforms during phagocytosis has not been addressed. The class I PI3Ks consist of a catalytic p110 isoform associated with a regulatory subunit. Mammals have three genes for the class IA p110 subunits encoding p110alpha, p110beta, and p110delta and one gene for the class IB p110 subunit encoding p110gamma. Here we report a specific recruitment of p110beta and p110delta (but not p110alpha) isoforms to the nascent phagosome during apoptotic cell phagocytosis by fibroblasts. By microinjecting inhibitory antibodies specific to class IA p110 subunits, we have shown that p110beta is the major isoform required for apoptotic cell and Fcgamma receptor-mediated phagocytosis by primary mouse macrophages. Macrophages from mice expressing a catalytically inactive form of p110delta showed no defect in the phagocytosis of apoptotic cells and IgG-opsonized particles, confirming the lack of a major role for p110delta in this process. Similarly, p110gamma-deficient macrophages phagocytosed apoptotic cells normally. Our findings demonstrate that p110beta is the major class I catalytic isoform required for apoptotic cell and Fcgamma receptor-mediated phagocytosis by primary macrophages.