Temperature-induced sign change of the magnetic interlayer coupling in Ni/Ni25Mn75/Ni trilayers on Cu3Au(001)

We investigated the magnetic interlayer coupling between two ferromagnetic (FM) Ni layers through an antiferromagnetic (AFM) Ni 25Mn75 layer and the influence of this coupling on the exchange bias phenomenon. The interlayer coupling energy of an epitaxial trilayer of 14 atomic monolayers (ML) Ni/45 ML Ni 25Mn75/16 ML Ni on Cu3Au(001) was extracted from minor-loop magnetization measurements using in-situ magneto-optical Kerr effect. The interlayer coupling changes from ferromagnetic to antiferromagnetic when the temperature is increased above 300 K. This sign change is interpreted as the result of the competition between an antiparallel Ruderman-Kittel-Kasuya-Yosida (RKKY)-type interlayer coupling, which dominates at high temperature, and a stronger direct exchange coupling across the AFM layer, which is present only below the Néel temperature of the AFM layer

Gutted adenoviral vector growth using E1/E2b/E3-deleted helper viruses

Background Helper-dependent, or gutted, adenoviruses (Ad) lack viral coding sequences, resulting in reduced immunotoxicity compared with conventional Ad vectors. Gutted Ad growth requires a conventional Ad to supply replication and packaging functions in trans . Methods that allow high-titer growth of gutted vectors while reducing helper contamination, and which use safer helper viruses, will facilitate the use of gutted Ad vectors in vivo . Methods Replication-defective helper viruses were generated that are deleted for Ad E1, E2b and E3 genes, but which contain lox P sites flanking the packaging signal. Complementing Ad packaging cell lines (C7-cre cells) were also generated by transfecting 293 cells with the Ad E2b genes encoding DNA polymerase and pre-terminal protein, and with a cre-recombinase plasmid. Results We show that C7-cre cells allow efficient production of gutted Ad using ΔE1 + ΔE2b + ΔE3 helper viruses whose growth can be limited by cre- lox P-mediated excision of the packaging signal. Gutted Ad vectors carrying ∼28 kb cassettes expressing full-length dystrophin were prepared at high titers, similar to those obtained with E2b+ helpers, with a resulting helper contamination of <1%. Conclusions These new packaging cell lines and helper viruses offer several significant advantages for gutted Ad vector production. They allow gutted virus amplification using a reduced number of passages, which should reduce the chances of selecting rearranged products. Furthermore, the residual helper contamination in gutted vector preparations should be less able to elicit immunological reactions upon delivery to tissues, since E2b-deleted vectors display a profound reduction in viral gene expression. Copyright © 2002 John Wiley & Sons, Ltd.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/35231/1/305_ftp.pd

Differentiated neuroprogenitor cells incubated with human or canine adenovirus, or lentiviral vectors have distinct transcriptome profiles

Several studies have demonstrated the potential for vector-mediated gene transfer to the brain. Helper-dependent (HD) human (HAd) and canine (CAV-2) adenovirus, and VSV-G-pseudotyped self-inactivating HIV-1 vectors (LV) effectively transduce human brain cells and their toxicity has been partly analysed. However, their effect on the brain homeostasis is far from fully defined, especially because of the complexity of the central nervous system (CNS). With the goal of dissecting the toxicogenomic signatures of the three vectors for human neurons, we transduced a bona fide human neuronal system with HD-HAd, HD-CAV-2 and LV. We analysed the transcriptional response of more than 47,000 transcripts using gene chips. Chip data showed that HD-CAV-2 and LV vectors activated the innate arm of the immune response, including Toll-like receptors and hyaluronan circuits. LV vector also induced an IFN response. Moreover, HD-CAV-2 and LV vectors affected DNA damage pathways - but in opposite directions - suggesting a differential response of the p53 and ATM pathways to the vector genomes. As a general response to the vectors, human neurons activated pro-survival genes and neuron morphogenesis, presumably with the goal of re-establishing homeostasis. These data are complementary to in vivo studies on brain vector toxicity and allow a better understanding of the impact of viral vectors on human neurons, and mechanistic approaches to improve the therapeutic impact of brain-directed gene transfer

ALCAM Regulates Motility, Invasiveness, and Adherens Junction Formation in Uveal Melanoma Cells

ALCAM, a member of the immunoglobulin superfamily, has been implicated in numerous developmental events and has been repeatedly identified as a marker for cancer metastasis. Previous studies addressing ALCAM’s role in cancer have, however, yielded conflicting results. Depending on the tumor cell type, ALCAM expression has been reported to be both positively and negatively correlated with cancer progression and metastasis in the literature. To better understand how ALCAM might regulate cancer cell behavior, we utilized a panel of defined uveal melanoma cell lines with high or low ALCAM levels, and directly tested the effects of manipulating these levels on cell motility, invasiveness, and adhesion using multiple assays. ALCAM expression was stably silenced by shRNA knockdown in a high-ALCAM cell line (MUM-2B); the resulting cells displayed reduced motility in gap-closure assays and a reduction in invasiveness as measured by a transwell migration assay. Immunostaining revealed that the silenced cells were defective in the formation of adherens junctions, at which ALCAM colocalizes with N-cadherin and ß-catenin in native cells. Additionally, we stably overexpressed ALCAM in a low-ALCAM cell line (MUM-2C); intriguingly, these cells did not exhibit any increase in motility or invasiveness, indicating that ALCAM is necessary but not sufficient to promote metastasis-associated cell behaviors. In these ALCAM-overexpressing cells, however, recruitment of ß-catenin and N-cadherin to adherens junctions was enhanced. These data confirm a previously suggested role for ALCAM in the regulation of adherens junctions, and also suggest a mechanism by which ALCAM might differentially enhance or decrease invasiveness, depending on the type of cadherin adhesion complexes present in tissues surrounding the primary tumor, and on the cadherin status of the tumor cells themselves

The role of the tissue microenvironment in the regulation of cancer cell motility and invasion

During malignant neoplastic progression the cells undergo genetic and epigenetic cancer-specific alterations that finally lead to a loss of tissue homeostasis and restructuring of the microenvironment. The invasion of cancer cells through connective tissue is a crucial prerequisite for metastasis formation. Although cell invasion is foremost a mechanical process, cancer research has focused largely on gene regulation and signaling that underlie uncontrolled cell growth. More recently, the genes and signals involved in the invasion and transendothelial migration of cancer cells, such as the role of adhesion molecules and matrix degrading enzymes, have become the focus of research. In this review we discuss how the structural and biomechanical properties of extracellular matrix and surrounding cells such as endothelial cells influence cancer cell motility and invasion. We conclude that the microenvironment is a critical determinant of the migration strategy and the efficiency of cancer cell invasion

Imaging magnetic responses of nanomagnets by XPEEM

The Spin-resolved Photoelectron Emission Microscope (SPEEM) is a permanently installed set-up at Helmholtz-Zentrum Berlin (HZB). Due to its specific contrast it is mainly used for magnetic imaging and micro-spectroscopy with quantitative analysis. A crucial point in magnetic imaging is the application of magnetic fields. Many experiments require observation of magnetic responses or the preparation of a certain magnetic state during the measurement. We present a dedicated magnetic sample holder combining magnetic field during imaging with additional temperature control. This set-up enables SPEEM to measure magnetization curves of individual Fe nanocubes (18 nm) 3 in size. If additionally alternating magnetic fields are applied we can image the local magnetic AC susceptibility (χAC) as a function of temperature. The latter is ideally suited to visualize local variations of the Curie temperature (TC) in nano- and microstructures. © 2012 Elsevier B.V.Support by F. Nolting is acknowledged for the design of the sample holder. J. H-A acknowledges financial support by MAT 2011/23791.Peer Reviewe