Defining the E-Cadherin repressor interactome in epithelial-mesenchymal transition : the PMC42 model as a case study

Epithelial-mesenchymal transition (EMT) is a feature of migratory cellular processes in all stages of life, including embryonic development and wound healing. Importantly, EMT features cluster with disease states such as chronic fibrosis and cancer. The dissolution of the E-cadherin-mediated adherens junction (AJ) is a key preliminary step in EMT and may occur early or late in the growing epithelial tumour. This is a first step for tumour cells towards stromal invasion, intravasation, extravasation and distant metastasis. The AJ may be inactivated in EMT by directed E-cadherin cleavage; however, it is increasingly evident that the majority of AJ changes are transcriptional and mediated by an expanding group of transcription factors acting directly or indirectly to repress E-cadherin expression. A review of the current literature has revealed that these factors may regulate each other in a hierarchical pattern where Snail1 (formerly Snail) and Snail2 (formerly Slug) are initially induced, leading to the activation of Zeb family members, TCF3, TCF4, Twist, Goosecoid and FOXC2. Within this general pathway, many inter-regulatory relationships have been defined which may be important in maintaining the EMT phenotype. This may be important given the short half-life of Snail1 protein. We have investigated these inter-regulatory relationships in the mesenchymal breast carcinoma cell line PMC42 (also known as PMC42ET) and its epithelial derivative, PMC42LA. This review also discusses several newly described regulators of E-cadherin repressors including oestrogen receptor-α and new discoveries in hypoxia- and growth factor-induced EMT. Finally, we evaluated how these findings may influence approaches to current cancer treatment.

Vimentin and epithelial-mesenchymal transition in human breast cancer : observations in vitro and in vivo

Breast cancer is a highly prevalent disease among women worldwide. While the expression of certain proteins within these tumours is used for prognosis and selection of therapies, there is a continuing need for additional markers to be identified. A considerable amount of current literature, based predominantly on cell culture systems, suggests that a major mechanism responsible for the progression of breast cancer is due to tumour cells losing their epithelial features and gaining mesenchymal properties. These events are proposed to be very similar to the epithelial-mesenchymal transition (EMT) process that has been well characterised in embryonic development. For the developmental and putative cancer EMT, the cell intermediate filament status changes from a keratin-rich network which connects to adherens junctions and hemidesmosomes, to a vimentin-rich network connecting to focal adhesions. This review summarises observations of vimentin expression in breast cancer model systems, and discusses the potential role of EMT in human breast cancer progression, and the prognostic usefulness of vimentin expression.

Tracking of physical activity, physical inactivity, and health-related physical fitness in rural youth

This study examined the tracking of selected measures of physical activity, inactivity, and fitness in a cohort of rural youth. Students (N = 181, 54.7% female, 63.5% African American) completed test batteries during their fifth-(age = 10.7 +/- 0.7 years), sixth-, and seventh-grade years. The Previous Day Physical Activity Recall (PDPAR) was used to assess 30-min blocks of vigorous physical activity (VPA), moderate-to-vigorous physical activity (MVPA), TV watching and other sedentary activities, and estimated energy expenditure (EE). Fitness measures included the PWC 170 cycle ergometer test, strength tests, triceps skinfold thickness, and BMI. Intraclass correlation coefficients (ICCs) for VPA, MVPA, and after-school EE ranged from 0.63 to 0.78. ICCs ranged from 0.49 to 0.71 for measures of inactivity and from 0.78 to 0.82 for the fitness measures. These results indicate that measures of physical activity, inactivity, and physical fitness tend to track during the transition from elementary to middle school.

Effect of constraint on ductile crack growth and ductile-brittle fracture transition of a carbon steel

Ductile-brittle fracture transition was investigated using compact tension (CT) specimens from -70oC to 40oC for a carbon steel. Large deformation finite element analysis has been carried out to simulate the stable crack growth in the compact tension (CT, a/W=0.6), three point-point bend (SE(B), a/W=0.1) and centre-cracked tension (M(T), a/W=0.5) specimens. Experimental crack tip opening displacement (CTOD) resistance curve was employed as the crack growth criterion. Ductile tearing is sensitive to constraint and tearing modulus increases with reduced constraint level. The finite element analysis shows that path-dependence of J-integral occurs from the very beginning of crack growth and ductile crack growth elevates the opening stress on the remaining ligament. Cleavage may occur after some ductile crack growth due to the increase of opening stress. For both stationary and growing cracks, the magnitude of opening stress increases with increasing in-plane constraint. The ductile-brittle transition takes place when the opening stress ahead of the crack tip reaches the local cleavage stress as the in-plane constraint of the specimen increases.

Remodeling of purinergic receptor-mediated Ca 2+ signaling as a consequence of EGF-induced epithelial-mesenchymal transition in breast cancer cells

Background The microenvironment plays a pivotal role in tumor cell proliferation, survival and migration. Invasive cancer cells face a new set of environmental challenges as they breach the basement membrane and colonize distant organs during the process of metastasis. Phenotypic switching, such as that which occurs during epithelial-mesenchymal transition (EMT), may be associated with a remodeling of cell surface receptors and thus altered responses to signals from the tumor microenvironment. Methodology/Principal Findings We assessed changes in intracellular Ca 2+ in cells loaded with Fluo-4 AM using a fluorometric imaging plate reader (FLIPR TETRA) and observed significant changes in the potency of ATP (EC 50 0.175 μM (-EGF) versus 1.731 μM (+EGF), P<0.05), and the nature of the ATP-induced Ca 2+ transient, corresponding with a 10-fold increase in the mesenchymal marker vimentin (P<0.05). We observed no change in the sensitivity to PAR2-mediated Ca 2+ signaling, indicating that these alterations are not simply a consequence of changes in global Ca 2+ homeostasis. To determine whether changes in ATP-mediated Ca 2+ signaling are preceded by alterations in the transcriptional profile of purinergic receptors, we analyzed the expression of a panel of P2X ionotropic and P2Y metabotropic purinergic receptors using real-time RT-PCR and found significant and specific alterations in the suite of ATP-activated purinergic receptors during EGF-induced EMT in breast cancer cells. Our studies are the first to show that P2X 5 ionotropic receptors are enriched in the mesenchymal phenotype and that silencing of P2X 5 leads to a significant reduction (25%, P<0.05) in EGF-induced vimentin protein expression. Conclusions The acquisition of a new suite of cell surface purinergic receptors is a feature of EGF-mediated EMT in MDA-MB-468 breast cancer cells. Such changes may impart advantageous phenotypic traits and represent a novel mechanism for the targeting of cancer metastasis.

The epithelial to mesenchymal transition and metastatic progression in carcinoma

Many data have emerged in support of the concept that the promotion of a migratory phenotype in epithelial cells is the result of an EMT, which leads to the loss of differentiation and to the acquisition of several mesenchymal features. Such an event is likely to occur during the metastatic conversion enabling the metastatic cell to invade the connective tissue and disseminate. Even though it cannot be excluded that some cancer cells might constitutively express a metastatic phenotype, the EMT implicated in the tumor progression process would rather be transient, induced in certain cells of the primary tumor by different factors, and reversed when the cells settle down in secondary organs (as schematically represented in Figure 1). However, it is clear that the phenotype that would emerge from the EMT occurring during tumor progression would also be modulated by the genetic background of the cells and must be to some extent different than the ones observed in normal physiological processes, and must also vary from one tumor to another.

Mesenchymal-epithelial transition (MET) as a mechanism for metastatic colonisation in breast cancer

As yet, there is no cure for metastatic breast cancer. Historically, considerable research effort has been concentrated on understanding the processes of metastasis, how a primary tumour locally invades and systemically disseminates using the phenotypic switching mechanism of epithelial to mesenchymal transition (EMT); however, much less is understood about how metastases are then formed. Breast cancer metastases often look (and may even function) as 'normal' breast tissue, a bizarre observation against the backdrop of the organ structure of the lung, liver, bone or brain. Mesenchymal to epithelial transition (MET), the opposite of EMT, has been proposed as a mechanism for establishment of the metastatic neoplasm, leading to questions such as: Can MET be clearly demonstrated in vivo? What factors cause this phenotypic switch within the cancer cell? Are these signals/factors derived from the metastatic site (soil) or expressed by the cancer cells themselves (seed)? How do the cancer cells then grow into a detectable secondary tumour and further disseminate? And finallyCan we design and develop therapies that may combat this dissemination switch? This review aims to address these important questions by evaluating long-standing paradigms and novel emerging concepts in the field of epithelial mesencyhmal plasticity.

A dual inverter-based supercapacitor direct integration scheme for wind energy conversion systems

Interfacing converters used in connecting energy storage systems like supercapacitors and battery banks to wind power systems introduce additional cost and power losses. This paper therefore presents a direct integration scheme for supercapacitors used in mitigating short-term power fluctuations in wind power systems. This scheme uses a dual inverter topology for both grid connection and interfacing a supercapacitor bank. The main inverter of the dual inverter system is powered by the rectified output of a wind turbine-coupled permanent-magnet synchronous generator. The auxiliary inverter is directly connected to the supercapacitor bank. With this approach, an interfacing converter is not required, and there are no associated costs and power losses incurred. The operation of the proposed system is discussed in detail. Simulation and experimental results are presented to verify the efficacy of the proposed system in suppressing short-term wind power fluctuations.

Design of a least-cost battery-supercapacitor energy storage system for realizing dispatchable wind power

A statistical approach is used in the design of a battery-supercapacitor energy storage system for a wind farm. The design exploits the technical merits of the two energy storage mediums, in terms of the differences in their specific power and energy densities, and their ability to accommodate different rates of change in the charging/discharging powers. By treating the input wind power as random and using a proposed coordinated power flows control strategy for the battery and the supercapacitor, the approach evaluates the energy storage capacities, the corresponding expected life cycle cost/year of the storage mediums, and the expected cost/year of unmet power dispatch. A computational procedure is then developed for the design of a least-cost/year hybrid energy storage system to realize wind power dispatch at a specified confidence level.

Modeling and position-sensorless control of a dual-airgap axial flux permanent magnet machine for flywheel energy storage systems

This paper presents the modeling and position-sensorless vector control of a dual-airgap axial flux permanent magnet (AFPM) machine optimized for use in flywheel energy storage system (FESS) applications. The proposed AFPM machine has two sets of three-phase stator windings but requires only a single power converter to control both the electromagnetic torque and the axial levitation force. The proper controllability of the latter is crucial as it can be utilized to minimize the vertical bearing stress to improve the efficiency of the FESS. The method for controlling both the speed and axial displacement of the machine is discussed. An inherent speed sensorless observer is also proposed for speed estimation. The proposed observer eliminates the rotary encoder, which in turn reduces the overall weight and cost of the system while improving its reliability. The effectiveness of the proposed control scheme has been verified by simulations and experiments on a prototype machine.

Diode-clamped three-level inverter-based battery/supercapacitor direct integration scheme for renewable energy systems

This paper describes a diode-clamped three-level inverter-based battery/supercapacitor direct integration scheme for renewable energy systems. The study is carried out for three different cases. In the first case, one of the two dc-link capacitors of the inverter is replaced by a battery bank and the other by a supercapacitor bank. In the second case, dc-link capacitors are replaced by two battery banks. In the third case, ordinary dc-link capacitors are replaced by two supercapacitor banks. The first system is supposed to mitigate both long-term and short-term power fluctuations while the last two systems are intended for smoothening long-term and short-term power fluctuations, respectively. These topologies eliminate the need for interfacing dc-dc converters and thus considerably improve the overall system efficiency. The major issue in aforementioned systems is the unavoidable imbalance in dc-link voltages. An analysis on the effects of unbalance and a space vector modulation method, which can produce undistorted current even in the presence of such unbalances, are presented in this paper. Furthermore, small vector selection-based power sharing and state of charge balancing techniques are proposed. Experimental results, obtained from a laboratory prototype, are presented to verify the efficacy of the proposed modulation and control techniques.

Direct integration of battery energy storage systems in distributed power generation

In this paper, a wind energy conversion system interfaced to the grid using a dual inverter is proposed. One of the two inverters in the dual inverter is connected to the rectified output of the wind generator while the other is directly connected to a battery energy storage system (BESS). This approach eliminates the need for an additional dc-dc converter and thus reduces power losses, cost, and complexity. The main issue with this scheme is uncorrelated dynamic changes in dc-link voltages that results in unevenly distributed space vectors. A detailed analysis on the effects of these variations is presented in this paper. Furthermore, a modified modulation technique is proposed to produce undistorted currents even in the presence of unevenly distributed and dynamically changing space vectors. An analysis on the battery charging/discharging process and maximum power point tracking of the wind turbine generator is also presented. Simulation and experimental results are presented to verify the efficacy of the proposed modulation technique and battery charging/discharging process.

Neutrophil gelatinase-associated lipocalin (NGAL) an early-screening biomarker for ovarian cancer : NGAL is associated with epidermal growth factor-induced epithelio-mesenchymal transition

The expression of neutrophil gelatinase-associated lipocalin (NGAL) has been shown to be upregulated in ovarian cancer cells. In this study, we report that the expression of immunoreactive NGAL (irNGAL) in ovarian tumors changes with disease grade and that this change is reflected in the concentration of NGAL in peripheral blood. A total of 59 ovarian tissues including normal, benign, borderline malignant and grades 1, 2 and 3 malignant were analyzed using immunohistochemistry. irNGAL was not present in normal ovaries and the NGAL expression was weak to moderate in benign tissues. Both borderline and grade 1 tumors displayed the highest amount of NGAL expression with moderate to strong staining, whereas in grade 2 and 3 tumors, the extent of staining was significantly less (p < 0.01) and staining intensity was weak to moderate. Staining in all cases was confined to the epithelium. NGAL expression was analyzed by ELISA in 62 serum specimens from normal and different grades of cancer patients. Compared to control samples, the NGAL concentration was 2 and 2.6-fold higher in the serum of patients with benign tumors and cancer patients with grade 1 tumors (p < 0.05) and that result was consistent with the expression of NGAL performed by Western blot. NGAL expression was evaluated by Western blot in an immortalized normal ovarian cell line (IOSE29) as well as ovarian cancer cell lines. Moderate to strong expression of NGAL was observed in epithelial ovarian cancer cell lines SKOV3 and OVCA433 while no expression of NGAL was evident in normal IOSE29 and mesenchyme-like OVHS1, PEO.36 and HEY cell lines. NGAL expression was downregulated in ovarian cancer cell lines undergoing epithelio-mesenchymal transition (EMT) induced by epidermal growth factor (EGF). Down-regulation of NGAL expression correlated with the upregulation of vimentin expression, enhanced cell dispersion and downregulation of E-cadherin expression, some of the hallmarks of EMT. EGF-induced EMT phenotypes were inhibited in the presence of AG1478, an inhibitor of EGF receptor tyrosine kinase activity. These data indicate that NGAL may be a good marker to monitor changes of benign to premalignant and malignant ovarian tumors and that the molecule may be involved in the progression of epithelial ovarian malignancies.

The epithelial-mesenchymal transition : new insights in signaling, development, and disease

The conversion of an epithelial cell to a mesenchymal cell is critical to metazoan embryogenesis and a de. ning structural feature of organ development. Current interest in this process, which is described as an epithelial- mesenchymal transition (EMT), stems from its developmental importance and its involvement in several adult pathologies. Interest and research in EMT are currently at a high level, as seen by the attendance at the recent EMT meeting in Vancouver, Canada (October 1-3, 2005). The meeting, which was hosted by The EMT International Association, was the second international EMT meeting, the . rst being held in Port Douglas, Queensland, Australia in October 2003. The EMT International Association was formed in 2002 to provide an international body for those interested in EMT and the reverse process, mesenchymal-epithelial transition, and, most importantly, to bring together those working on EMT in development, cancer, . brosis, and pathology. These themes continued during the recent meeting in Vancouver. Discussion at the Vancouver meeting spanned several areas of research, including signaling pathway activation of EMT and the transcription factors and gene targets involved. Also covered in detail was the basic cell biology of EMT and its role in cancer and . brosis, as well as the identi. cation of new markers to facilitate the observation of EMT in vivo. This is particularly important because the potential contribution of EMT during neoplasia is the subject of vigorous scientific debate (Tarin, D., E.W. Thompson, and D.F. Newgreen. 2005. Cancer Res. 65:5996-6000; Thompson, E.W., D.F. Newgreen, and D. Tarin. 2005. Cancer Res. 65:5991-5995).

Cadherins in the human placenta : epithelial-mesenchymal transition (EMT) and placental development

Colonisation of the maternal uterine wall by the trophoblast involves a series of alterations in the behaviour and morphology of trophoblast cells. Villous cytotrophoblast cells change from a well-organised coherently layered phenotype to one that is extravillous, acquiring a proliferative, migratory and invasive capacity, to facilitate fetal-maternal interaction. These changes are similar to those of other developmental processes falling under the umbrella of an epithelial-mesenchymal transition (EMT). Modulation of cell adhesion and cell polarity occurs through changes in cell-cell junctional molecules, such as the cadherins. The cadherins, particularly the classical cadherins (e.g. Epithelial-(E)-cadherin), and their link to adaptors called catenins at cell-cell contacts, are important for maintaining cell attachment and the layered phenotype of the villous cytotrophoblast. In contrast, reduced expression and re-organization of cadherins from these cell junctional regions promote a loosened connection between cells, coupled with reduced apico-basal polarity. Certain non-classical cadherins play an active role in cell migration processes. In addition to the classical cadherins, two other cadherins which have been reported in placental tissues are vascular endothelial (VE) cadherin and cadherin-11. Cadherin molecules are well placed to be key regulators of trophoblast cell behaviour, analogous to their role in other developmental EMTs. This review addresses cadherin expression and function in normal and diseased human placental tissues, especially in fetal growth restriction and pre-eclampsia where trophoblast invasion is reduced.