Biological and molecular characterization of a resistance-breaking isolate of citrus tristeza virus from Uruguay and its effects on Poncirus trifoliata growth performance.

Resistance-breaking (RB) isolates of citrus tristeza virus (CTV) can replicate and move systemically in Poncirus trifoliata, a rootstock widely used for management of decline caused by CTV and other purposes. In Uruguay, severe CTV isolates are prevalent, and an RB isolate (designated as RB-UY1) was identified. In order to predict the implications of this genotype circulating in citrus crops grafted on trifoliate rootstocks, the aim of this work was to determine the biological and molecular characteristics of this isolate, the efficiency of its transmission by Toxoptera citricida, and its effects on plant growth performance of P. trifoliata. Our results show that RB-UY1 can be classified as a mild isolate, that it is phylogenetically associated with the RB1 group, and that it is efficiently transmitted by T. citrida. They also suggest that the RB-UY1 isolate should not affect the performance of citrus crops grafted on trifoliate rootstocks, although some growth parameters of P. trifoliata seedlings were affected four years after inoculation.

Self-renewal of human embryonic stem cells requires insulin-like growth factor-1 receptor and ERBB2 receptor signaling.

Despite progress in developing defined conditions for human embryonic stem cell (hESC) cultures, little is known about the cell-surface receptors that are activated under conditions supportive of hESC self-renewal. A simultaneous interrogation of 42 receptor tyrosine kinases (RTKs) in hESCs following stimulation with mouse embryonic fibroblast (MEF) conditioned medium (CM) revealed rapid and prominent tyrosine phosphorylation of insulin receptor (IR) and insulin-like growth factor-1 receptor (IGF1R); less prominent tyrosine phosphorylation of epidermal growth factor receptor (EGFR) family members, including ERBB2 and ERBB3; and trace phosphorylation of fibroblast growth factor receptors. Intense IGF1R and IR phosphorylation occurred in the absence of MEF conditioning (NCM) and was attributable to high concentrations of insulin in the proprietary KnockOut Serum Replacer (KSR). Inhibition of IGF1R using a blocking antibody or lentivirus-delivered shRNA reduced hESC self-renewal and promoted differentiation, while disruption of ERBB2 signaling with the selective inhibitor AG825 severely inhibited hESC proliferation and promoted apoptosis. A simple defined medium containing an IGF1 analog, heregulin-1beta (a ligand for ERBB2/ERBB3), fibroblast growth factor-2 (FGF2), and activin A supported long-term growth of multiple hESC lines. These studies identify previously unappreciated RTKs that support hESC proliferation and self-renewal, and provide a rationally designed medium for the growth and maintenance of pluripotent hESCs.

Distribution of thrombospondin-4 in the bovine eye.

PURPOSE: The purpose of this study was to analyze the distribution of thrombospondin-4 (TSP-4) in the bovine eye. METHODS: Anterior and posterior segments of the bovine eyes were sectioned and stained by the indirect immunofluorescence method with an anti-TSP-4 antibody. The tissues were analyzed by reverse-transcription-polymerase chain reaction (RT-PCR) to determine where the TSP-4 mRNA is produced. RESULTS: Immunohistochemical staining for TSP-4 indicated the presence of TSP-4 in the cornea (epithelium, basement membrane, and keratocytes), conjunctiva (epithelium and stroma), aqueous ducts, sclera, iris (stroma), ciliary processes and muscle, trabecular meshwork, Bruch's membrane, retina, lamina cribrosa, and optic nerve, and in all blood vessel walls. TSP-4 mRNA was expressed by the cells in all structures. CONCLUSIONS: TSP-4 is widely distributed in the bovine eye where it may play a role in the functions of basement membranes in various tissues. It is abundant in the trabecular and uveo-scleral pathways and may play a role in the regulation of aqueous outflow resistance.