Western Immunoblotting in Avian Shell Gland Sample Immunoblotting Methods1

Avian shell gland tissue was subjected to Western blot analysis using anti-human estrogen receptor- α antibody H222. Initial attempts to obtain consistent, high-quality blots were unsuccessful because, as it turned out, excessive lipid in tissue preparations interfered with protein separation. Incremental additions of acetone eventually proved to be the critical step in solubilizing lipids and allowing consistent separation of bands on gels. A detailed description of the methodology is presented

Estrogen Receptor-α Populations Change with Age in Commercial Laying Hens

Older hens in production lay larger but fewer eggs than younger birds, and the incidence of soft and broken shells is greater in older hens than younger. These changes are attributable at least in part to changing hormone profiles and diminished ability of the hen to transport calcium at the duodenum. In further exploration of this relationship, a study was conducted with three ages of Hy-Line W-36 birds: prelay pullets (PL; 19 wk, 0% production), peak-production hens (PP; 29 wk, ∼93% production), and late-stage hens (LS; 71 wk, ∼80% production). Hens from the PP and LS groups were palpated for presence of an egg in the shell gland; hens were then euthanized and tissues (kidney, shell gland, hypothalamus) were removed for quantification of estrogen receptor- α (ERα) populations via immunocytochemical and Western blot analyses. Localization of ERα by immunostaining in the shell gland showed differences among age groups; however, no differences were noted in localization of ERα between age groups in the kidney and hypothalamus. In both the kidney and the shell gland there was a decrease in the amount of ERα, as detected by immunoblotting, in the LS hens compared to PL and PP birds (P \u3c 0.05). The results suggest that failure of calcium regulating mechanisms with age may be mediated at least in part through the reduced populations of estrogen receptors in certain critical tissues

Contact-dependent killing by cytotoxic T lymphocytes is insufficient for EL4 tumor regression in vivo

Immunotherapies are an emerging strategy for treatment of solid tumors. Improved understanding of the mechanisms employed by cytotoxic T lymphocytes (CTLs) to control tumors will aid in the development of immunotherapies. CTLs can directly kill tumor cells in a contact-dependent manner or may exert indirect effects on tumor cells via secretion of cytokines. Here we aim to quantify the importance of these mechanisms in murine thymoma EL4/EG7 cells. We developed an agent-based model (ABM) and an ordinary differential equation (ODE) model of tumor regression after adoptive transfer of a population of CTLs. Models were parameterized based on in vivo measurements of CTL infiltration and killing rates applied to EL4/EG7 tumors and OTI T cells. We quantified whether infiltrating CTLs are capable of controlling tumors through only direct, contact-dependent killing. Both models agreed that the low measured killing rate of CTLs in vivo was insufficient to cause tumor regression. In our ABM we also simulated CTL production of the cytokine interferon gamma (IFNγ) in order to explore how an antiproliferative effect of IFNγ might aid CTLs in tumor control. In this model IFNγ substantially reduced tumor growth compared to direct killing alone. Collectively these data demonstrate that contact-dependent killing is insufficient for EL4 regression in vivo and highlight the potential importance of cytokine-induced antiproliferative effects in T cell-mediated tumor control.Toxicolog

Heterogeneous, delayed-onset killing by multiple-hitting T cells: Stochastic simulations to assess methods for analysis of imaging data

Although quantitative insights into the killing behaviour of Cytotoxic T Lymphocytes (CTLs) are necessary for the rational design of immune-based therapies, CTL killing function remains insufficiently characterised. One established model of CTL killing treats CTL cytotoxicity as a Poisson process, based on the assumption that CTLs serially kill antigen-presenting target cells via delivery of lethal hits, each lethal hit corresponding to a single injection of cytotoxic proteins into the target cell cytoplasm. Contradicting this model, a recent in vitro study of individual CTLs killing targets over a 12-hour period found significantly greater heterogeneity in CTL killing performance than predicted by Poisson-based killing. The observed killing process was dynamic and varied between CTLs, with the best performing CTLs exhibiting a marked increase in killing during the final hours of the experiments, along with a "burst killing" kinetic. Despite a search for potential differences between CTLs, no mechanistic explanation for the heterogeneous killing kinetics was found. Here we have used stochastic simulations to assess whether target cells might require multiple hits from CTLs before undergoing apoptosis, in order to verify whether multiple-hitting could explain the late onset, burst killing dynamics observed in vitro. We found that multiple-hitting from CTLs was entirely consistent with the observed killing kinetics. Moreover, the number of available targets and the spatiotemporal kinetics of CTL:target interactions influenced the realised CTL killing rate. We subsequently used realistic, spatial simulations to assess methods for estimating the hitting rate and the number of hits required for target death, to be applied to microscopy data of individual CTLs killing targets. We found that measuring the cumulative duration of individual contacts that targets have with CTLs would substantially improve accuracy when estimating the killing kinetics of CTLs.Toxicolog

Search for a T_20 Analyzer for Deuterons

This work was supported by the National Science Foundation Grant NSF PHY 81-14339 and by Indiana Universit

Gemini Observations of Disks and Jets in Young Stellar Objects and in Active Galaxies

We present first results from the Near-infrared Integral Field Spectrograph (NIFS) located at Gemini North. For the active galaxies Cygnus A and Perseus A we observe rotationally-supported accretion disks and adduce the existence of massive central black holes and estimate their masses. In Cygnus A we also see remarkable high-excitation ionization cones dominated by photoionization from the central engine. In the T-Tauri stars HV Tau C and DG Tau we see highly-collimated bipolar outflows in the [Fe II] 1.644 micron line, surrounded by a slower molecular bipolar outflow seen in the H_2 lines, in accordance with the model advocated by Pyo et al. (2002).Comment: Invited paper presented at the 5th Stromlo Symposium. 9 pages, 7 figures. Accepted for publication in Astrophysics & Space Scienc

Procedures for the salvage and necropsy of the Dugong (Dugong Dugon)-second edition 2007

This manual provides a detailed guide for dugong (Dugong dugon) carcass handling and necropsy procedures. It is intended to be used as a resource and training guide for anyone involved in dugong incidents including management officers, biologists, parks and wildlife field staff, and veterinarians and pathologists who may lack dugong expertise. Because of the wide range of professionals this book is targeting, information and the use of technical terms is extensive. Section 8.0 provides definitions of various terms used which are italicised throughout the text

Deformation effects in 56^{56}Ni nuclei produced in 28^{28}Si+28^{28}Si at 112 MeV

Velocity and energy spectra of the light charged particles (protons and $\alpha$-particles) emitted in the $^{28}$Si(E$_{lab}$ = 112 MeV) + $^{28}$Si reaction have been measured at the Strasbourg VIVITRON Tandem facility. The ICARE charged particle multidetector array was used to obtain exclusive spectra of the light particles in the angular range 15 - 150 degree and to determine the angular correlations of these particles with respect to the emission angles of the evaporation residues. The experimental data are analysed in the framework of the statistical model. The exclusive energy spectra of $\alpha$-particles emitted from the $^{28}$Si + $^{28}$Si compound system are generally well reproduced by Monte Carlo calculations using spin-dependent level densities. This spin dependence approach suggests the onset of large deformations at high spin. A re-analysis of previous $\alpha$-particle data from the $^{30}$Si + $^{30}$Si compound system, using the same spin-dependent parametrization, is also presented in the framework of a general discussion of the occurrence of large deformation effects in the A$_{CN}$ ~ 60 mass region.Comment: 25 pages, 6 figure