Grainyhead genes are involved in wound healing and developmental neural tube closure. In light of the high degree of similarity between the epithelial-mesenchymal transitions (EMT) occurring in wound healing processes and the cancer stem cell-like compartment of tumors, including TGF-β-dependence, we investigated the role of the Grainyhead gene, Grainyhead-Like-2 (GRHL2) in oncogenic EMT. GRHL2 was down-regulated specifically in the claudin-low subclass breast tumors and in basal-B subclass breast cancer cell lines. GRHL2 suppressed TGF-β-induced, Twist-induced or spontaneous EMT, enhanced anoikis-sensitivity, and suppressed mammosphere generation in mammary epithelial cells. These effects were mediated in part by suppression of ZEB1 expression via direct repression of the ZEB1 promoter. GRHL2 also inhibited Smad-mediated transcription and it upregulated mir200b/c as well as the TGF-β receptor antagonist, BMP2. Lastly, ectopic expression of GRHL2 in MDA-MB-231 breast cancer cells triggered a mesenchymal-to-epithelial transition and restored sensitivity to anoikis. Taken together, our findings define a major role for GRHL2 in the suppression of oncogenic EMT in breast cancer cells.

Stability under dynamical changes to network connectivity is invoked alongside previous criteria to constrain brain network architecture. A new hierarchical network is introduced that satisfies all these constraints, unlike more commonly studied regular, random, and small-world networks. It is shown that hierarchical networks can simultaneously have high clustering, short path lengths, and low wiring costs, while being robustly stable under large scale reconnection of substructures.

Anoikis-resistance of tumor cells is critical for anchorage-independent growth and metastasis. The inflammatory-response transcription factor NF-κB contributes to anoikis-resistance and tumor progression through mechanisms that are understood incompletely. Deleted in Breast Cancer-1 protein (KIAA1967) is over-expressed in several tumor types, and correlates with a poorer prognosis in some cases. We report here that DBC1 suppressed anoikis in normal epithelial and breast cancer cell lines. DBC1 interacted with IKK-β, stimulating its kinase activity, promoting NF-κB transcriptional activity through the phosphorylation of relA serine-536 and enhancing the expression of the NF-κB target genes, c-FLIP and bcl-xl. Our results indicate that DBC1 is an important co-factor for the control of the IKK-β-NF-κB signaling pathway that regulates anoikis.

A previous report from this laboratory (1) presented the results of hematologic studies in nine dogs cooled below 200 C. During induction of hypothermia the animals showed a marked rise in red cell count, hemoglobin, and hematocrit, together with a nearly complete disappearance of white cells and platelets. On rewarming, the values all returned to normal. A number of studies have been carried out to learn more about the mechanism of these changes. The present paper describes some of the studies dealing especially with the changes in platelets and white cells. MATERIALS AND METHODSThe experiments were performed on a group of 22 adult mongrel dogs. Hypothermia was induced by a method of surface cooling previously reported (2). Each dog was anesthetized with intravenous thiopental. An endotracheal catheter was inserted and connected to an automatic respirator utilizing room air. Thermocouples in the rectum and esophagus permitted continuous temperature recordings by a Brown potentiometer. A continuous three limb lead electrocardiogram and aortic blood pressure were also recorded. The animal was placed in an ice bath at 40 C. and cooled until ventricular fibrillation occurred or the rectal temperature reached 170 C. The heart was defibrillated electrically.Each experiment, from the onset of hypothermia until rewarming had raised the body temperature to approximately 30°C., occupied approximately five hours. Platelet counts were carried out by the method of Brecher and Cronkite (3) using phase microscopy. Two pipettes were filled and two chambers for each pipette were counted and the average was taken as the result for each determination. By this technique, the normal range for platelet counts in dogs is 200,000 to 350,000 per cu. mm. Leukocyte counts were carried out by the standard hemocytometer technique using 1: 20 dilution. Both sides of two hemocytometer chambers were counted. Hemoglobin concentration was measured by the cyanmethemoglobin method (4). Hematocrit was measured in capillary tubes after high-speed centrifugation (5).Protocol 1: Bone marrow aspirations and biopsies were carried out from the ribs and tibias of three dogs before, during, and after hypothermia.Protocol 2: Studies with radioactive platelets were carried out in dogs using a donor (Dog A) and a recipient (Dog B). Two mc. of Pa (NaHP"0O, Abbott) was injected into Dog A. One week later, Dog A was bled by cardiac puncture, and 500 ml. of blood was drawn into a plastic bag containing disodium versinate solution. Simultaneously 500 ml. of blood was taken from Dog B. Dog B was then transfused immediately with the blood from Dog A containing radioactive platelets. Onehalf hour later, using siliconized equipment and a two syringe technique, 15 ml. of blood was taken from Dog B. Platelets and plasma were separated from red cells by differential centrifugation and the platelets were counted in a hemocytometer chamber using a phasecontrast microscope. The platelet suspension was then centrifuged to throw down the platelets. The platelets were washed ...

Statistical relationships between tornado intensity and various stability and wind-related parameters are determined from a dataset of more than 200 tornado proximity soundings. Soundings were obtained from two sources: the University of Missouri and from analyses. The surface to 600 hPa wind shear, streamwise vorticity, storm velocity, and storm-relative environmental helicity are shown to be best correlated with tornado intensity. A multiple regression analysis produced an equation relating tornado intensity to the lifted index and the surface to 600 hPa wind shear. This relationship could be used operationally to predict tornado intensity. Mean temperature , dewpoint temperature, and wind profiles are presented for each tornado intensity category and these may be useful as input to numerical thunderstorm simulations. Progressively cooler temperatures and increasing wind speed and vertical wind shear are evident with increasing tornado intensity. A modified form of the bulk Richardson number is derived to assist the forecasting of isolated supercell thunderstorm occurrence. The modified form uses parameters that are readily available to the operational forecaster.