Increased ovulation rate in virgin mice induced by males

No abstract availabl

Temporally resolved interactions between antigen-stimulated IgE receptors and Lyn kinase on living cells

Upon cross-linking by antigen, the high affinity receptor for immunoglobulin E (IgE), FcɛRI, is phosphorylated by the Src family tyrosine kinase Lyn to initiate mast cell signaling, leading to degranulation. Using fluorescence correlation spectroscopy (FCS), we observe stimulation-dependent associations between fluorescently labeled IgE-FcɛRI and Lyn-EGFP on individual cells. We also simultaneously measure temporal variations in the lateral diffusion of these proteins. Antigen-stimulated interactions between these proteins detected subsequent to the initiation of receptor phosphorylation exhibit time-dependent changes, suggesting multiple associations between FcɛRI and Lyn-EGFP. During this period, we also observe a persistent decrease in Lyn-EGFP lateral diffusion that is dependent on Src family kinase activity. These stimulated interactions are not observed between FcɛRI and a chimeric EGFP that contains only the membrane-targeting sequence from Lyn. Our results reveal real-time interactions between Lyn and cross-linked FcɛRI implicated in downstream signaling events. They demonstrate the capacity of FCS cross-correlation analysis to investigate the mechanism of signaling-dependent protein–protein interactions in intact, living cells

and Stable Core-Shell Fluorescence Silica Nanoparticles

ABSTRACT A class of highly fluorescent and photostable core−shell nanoparticles from a modified Sto 1ber synthesis in the size range of 20−30 nm is described. These nanoparticles are monodisperse in solution, 20 times brighter, and more photostable than their constituent fluorophore, and are amenable to specific labeling of biological macromolecules for bioimaging experiments. The photophysical characteristics of the encapsulated fluorophores differ from their solution properties. This raises the possibility of tuning nanoparticle structure toward enhanced radiative properties, making them an attractive material platform for a diverse range of applications. Fluorescent nanoparticles have tremendous promise as indicators and photon sources for a number of biotechnological and information technology applications such as biological imaging, sensor technology, microarrays, and optical computing. 1 These applications all require size-controlled, monodisperse, bright nanoparticles that can be specifically conjugated to biological macromolecules or arranged in higherorder structures. Nanoparticles with core-shell architecture have the added benefit of providing a robust platform for incorporating diverse functionalities into a single nanoparticle. 2 We have developed a novel class of multifunctional silicabased fluorescent nanoparticles using a synthesis based on the Stöber method. The Stöber synthesis of colloidal silica, by which monodisperse nano-to micrometer sized silica particles may be obtained, was first described in 1968. 3 Van Blaaderen and co-workers first reported the covalent incorporation of organic fluorophores into Stöber colloidal silica and the synthesis of fluorescent silica nanoparticles in the hundreds of nanometers size range. Fluorescence correlation spectroscopy (FCS) is used as a primary investigative tool to characterize these nanoparticles. FCS has now become a common tool for characterizing the properties of fluorescent moieties in solution

Correlation functions quantify super-resolution images and estimate apparent clustering due to over-counting

We present an analytical method to quantify clustering in super-resolution localization images of static surfaces in two dimensions. The method also describes how over-counting of labeled molecules contributes to apparent self-clustering and how the effective lateral resolution of an image can be determined. This treatment applies to clustering of proteins and lipids in membranes, where there is significant interest in using super-resolution localization techniques to probe membrane heterogeneity. When images are quantified using pair correlation functions, the magnitude of apparent clustering due to over-counting will vary inversely with the surface density of labeled molecules and does not depend on the number of times an average molecule is counted. Over-counting does not yield apparent co-clustering in double label experiments when pair cross-correlation functions are measured. We apply our analytical method to quantify the distribution of the IgE receptor (Fc{\epsilon}RI) on the plasma membranes of chemically fixed RBL-2H3 mast cells from images acquired using stochastic optical reconstruction microscopy (STORM) and scanning electron microscopy (SEM). We find that apparent clustering of labeled IgE bound to Fc{\epsilon}RI detected with both methods arises from over-counting of individual complexes. Thus our results indicate that these receptors are randomly distributed within the resolution and sensitivity limits of these experiments.Comment: 22 pages, 5 figure

Short-term change in growth of uterine leiomyoma: tumor growth spurts

To describe the short-term changes in growth of uterine leiomyomata (fibroids)

Inhibition of Ets-1 DNA Binding and Ternary Complex Formation between Ets-1, NF-kappa B, and DNA by a Designed DNA-binding Ligand

Sequence-specific pyrrole-imidazole polyamides can be designed to interfere with transcription factor binding and to regulate gene expression, both in vitro and in living cells. Polyamides bound adjacent to the recognition sites for TBP, Ets-1, and LEF-1 in the human immunodeficiency virus, type 1 (HIV-1), long terminal repeat inhibited transcription in cell-free assays and viral replication in human peripheral blood lymphocytes. The DNA binding activity of the transcription factor Ets-1 is specifically inhibited by a polyamide bound in the minor groove. Ets-1 is a member of the winged-helix-turn-helix family of transcription factors and binds DNA through a recognition helix bound in the major groove with additional phosphate contacts on either side of this major groove interaction. The inhibitory polyamide possibly interferes with phosphate contacts made by Ets-1, by occupying the adjacent minor groove. Full-length Ets-1 binds the HIV-1 enhancer through cooperative interactions with the p50 subunit of NF-kappa B, and the Ets-inhibitory polyamide also blocks formation of ternary Ets-1·NF-kappa B·DNA complexes on the HIV-1 enhancer. A polyamide bound adjacent to the recognition site for NF-kappa B also inhibits NF-kappa B binding and ternary complex formation. These results broaden the application range of minor groove-binding polyamides and demonstrate that these DNA ligands are powerful inhibitors of DNA-binding proteins that predominantly use major groove contacts and of cooperative protein-DNA ternary complexes

Issues in Research on the Young Chronically III Child

A major goal of research on chronic illness in children is to determine how the illness interacts with developmental processes. The child must be studied within the context of the family, the school, and the health care system. Problems in research include the use of appropriate control groups and matching on control variables. The generic, or cross-categorical, approach has led to the identification of factors affecting children regardless of particular illness. Adjustment to school depends on coordination of the family and health professionals with personnel within the school.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/68724/2/10.1177_027112148600500406.pd