The Role of MCP-1 in Epithelial to Mesenchymal Transition and Invasion during Early Mammary Carcinogenesis

<p>Triple negative breast cancers are aggressive breast tumors that are characterized by the absence of estrogen receptor, progesterone receptor, and HER2. This absence of expression limits targeted therapeutic strategies. The result is a disease that is associated with a poor prognosis. The current survival rate for those afflicted with triple negative breast cancer is 77%. This survival rate is compared to a 93% survival rate for all other breast cancers combined. Furthermore, triple negative breast cancers disproportionately affect young and African American women. </p><p>The Carolina Breast Cancer Study demonstrated that obesity increased the risk of triple negative breast cancer in premenopausal women. These findings were supported by a meta-analysis of 11 epidemiological studies evaluating the association between triple negative breast cancer, obesity, and menopause status. Consequently, it is hypothesized that obesity in premenopausal women may contribute to the initiation and progression of triple negative breast cancer. </p><p>On average, adipose constitutes 70% of the breast. Obesity is a disease of increased adipose tissue, which can increase the adiposity of the breast. Studies show that obesity enhances the production of the inflammatory cytokines IL1&#946;, IL6, IL8, TNF&#945;, and MCP-1 in adipose tissue. Inflammatory cytokines contribute to proliferation and survival of malignant cells, migration and invasion, and induction of epithelial to mesenchymal transition (EMT). </p><p>The purpose of this study is to investigate the role of adipose derived cytokines in promotion of EMT and invasion in early mammary carcinogenesis. To study EMT and invasion in early mammary carcinogenesis, a co-culture was generated using adipose stromal cell (ASC) conditioned media and the non-transformed, mammary epithelial cell line MCF10A. ASC-conditioned media was characterized for cytokine production. Findings indicated that IL6, IL8, and MCP-1 were secreted into the media. Researchers have extensively studied IL6 and IL8 in breast EMT, but the relevance of MCP-1 in breast carcinogenesis has only recently emerged. MCP-1 is involved in monocyte and macrophage trafficking, renal fibrosis and EMT, as well as EMT in peritoneal disease. The role of MCP-1 in renal and peritoneal EMT suggests that MCP-1 may induce EMT in mammary epithelial cells. </p><p>MCP-1 did not alter the protein expression levels of either E-cadherin or Vimentin in 2D culture. MCP-1 treatment did, however, rapidly phosphorylate Erk1/2 at T202/Y204 and induce chemotaxis of MCF10A cells in a Boyden Chamber migration assay. Erk1/2 phosphorylation and chemotaxis were inhibited by pretreatment with PD98059, a potent small molecule inhibitor of Erk signaling. </p><p>In 3D, MCF10A acini treated with MCP-1 lost expression of E-cadherin, &#946;-catenin, and localized areas of Integrin &#945;6. Additionally, MCP-1 induced outgrowth away from the MCF10A spheroids and invasion into the extracellular matrix during a spheroid gel invasion assay. MCP-1 did not induce the secretion of MMP-2 or MMP-9 during invasion. When the MCP-1 treated acini and spheroids were co-treated with PD98059 E-cadherin loss and invasion were inhibited respectively. This inhibition suggests that Erk activation is necessary for E-cadherin loss and cell invasion. The data indicate that MCP-1 plays a potential role in the early EMT and invasion of non-transformed, mammary epithelial cells. This study provides a foundation for the study of MCP-1 induced EMT and invasion in MCF10A cells. Further work will need to be completed to elucidate the mechanism by which MCP-1 decreases E-cadherin and induces invasion.</p>Thesi

VANG-1 and PRKL-1 Cooperate to Negatively Regulate Neurite Formation in Caenorhabditis elegans

Neuritogenesis is a critical early step in the development and maturation of neurons and neuronal circuits. While extracellular directional cues are known to specify the site and orientation of nascent neurite formation in vivo, little is known about the genetic pathways that block inappropriate neurite emergence in order to maintain proper neuronal polarity. Here we report that the Caenorhabditis elegans orthologues of Van Gogh (vang-1), Prickle (prkl-1), and Dishevelled (dsh-1), core components of planar cell polarity (PCP) signaling, are required in a subset of peripheral motor neurons to restrict neurite emergence to a specific organ axis. In loss-of-function mutants, neurons display supernumerary neurites that extend inappropriately along the orthogonal anteroposterior (A/P) body axis. We show that autonomous and non-autonomous gene activities are required early and persistently to inhibit the formation or consolidation of growth cone protrusions directed away from organ precursor cells. Furthermore, prkl-1 overexpression is sufficient to suppress neurite formation and reorient neuronal polarity in a vang-1– and dsh-1–dependent manner. Our findings suggest a novel role for a PCP–like pathway in maintaining polarized neuronal morphology by inhibiting neuronal responses to extrinsic or intrinsic cues that would otherwise promote extraneous neurite formation

Midnight Forest

Meet a raccoon searching for food, a fox chasing bats, dragons waking for adventure, and more! This rhyming book is designed with the visually impaired in mind, and for all to enjoy.https://digitalcommons.cedarville.edu/alum_books/1502/thumbnail.jp

Compliance with reduced-impact harvesting guidelines by timber enterprises in terra firme forests of the Brazilian Amazon

The paper presents the results and main conclusions of an assessment of compliance with technical guidelines for Reduced Impact Harvesting (RIH) in terra firme forests of the Brazilian Amazon. The assessment was carried out in two certified timber enterprises in the State of Para, Brazil applying the RIH-guidelines for a period of over three years. From a tool developed for Amazonian forest enterprises to monitor the sustainability of their harvesting operations, which uses a set of criteria and indicators (C&I), a total of 190 verifiers were selected for assessing the 61 RIH-guidelines. The assessment revealed valuable information with regard to the state of implementation and quality of the forest operations in the two enterprises and important insights for improvement of the RIH-guidelines. Two thirds of the RIH-guidelines were fully implemented. Their acceptance, however, differed according to the situation and interest of the enterprises. Among the reasons for incomplete implementation of the RIH-guidelines, the lack of systematic monitoring, insufficient training and qualification, and inadequate equipment appeared to be most important. The study also showed the need for the continuous assessment of the quality and relevance of RIH-guidelines

Targeted development of registries of biological parts

The design and construction of novel biological systems by combining basic building blocks represents a dominant paradigm in synthetic biology. Creating and maintaining a database of these building blocks is a way to streamline the fabrication of complex constructs. The Registry of Standard Biological Parts (Registry) is the most advanced implementation of this idea

CD and NMR conformational studies of a peptide encompassing the Mid Loop interface of Ship2-Sam

The lipid phosphatase Ship2 is a protein that intervenes in several diseases such as diabetes, cancer, neurodegeneration, and atherosclerosis. It is made up of a catalytic domain and several protein docking modules such as a C-terminal Sam (Sterile alpha motif) domain. The Sam domain of Ship2 (Ship2-Sam) binds to the Sam domains of the EphA2 receptor (EphA2-Sam) and the PI3K effector protein Arap3 (Arap3-Sam). These heterotypic Sam-Sam interactions occur through formation of dimers presenting the canonical "Mid Loop/End Helix" binding mode. The central region of Ship2-Sam, spanning the C-terminal end of α2, the α3 and α4 helices together with the α2α3 and α3α4 interhelical loops, forms the Mid Loop surface that is needed to bind partners Sam domains. A peptide encompassing most of the Ship2-Sam Mid Loop interface (Shiptide) capable of binding to both EphA2-Sam and Arap3-Sam, was previously identified. Here we investigated the conformational features of this peptide, through solution CD and NMR studies in different conditions. These studies reveal that the peptide is highly flexible in aqueous buffer, while it adopts a helical conformation in presence of 2,2,2-trifluoroethanol. The discovered structural insights and in particular the identification of a helical motif, may lead to the design of more constrained and possibly cell permeable Shiptide analogs that could work as efficient antagonists of Ship2-Sam heterotypic interactions and embrace therapeutic applications. © 2014 Wiley Periodicals, Inc. Biopolymers 101: 1088-1098, 2014. © 2014 Wiley Periodicals, Inc