Integrin alpha IIb promoter-targeted expression of gene products in megakaryocytes derived from retrovirus-transduced human hematopoietic cells

Megakaryocyte-specific expression of the platelet-adhesion receptor, integrin alphaIIbbeta3, is caused by the presence of regulatory elements of the alphaIIb promoter that direct high-level, selective gene transcription early in megakaryocytopoiesis. To develop methods for targeted expression of transgenes, we transduced human CD34+ peripheral blood cells with a murine leukemia virus (MuLV) vector controlled by the human integrin alphaIIb promoter (nucleotides -889 to +35). A naturally occurring cDNA encoding the Pl(A2) alloantigen form (Pro(33)) of the integrin beta3 subunit was subcloned into this construct (-889Pl(A2)beta3) and transduced into cells that endogenously synthesized Pl(A1)beta3 (Leu(33)) as a marker for detection of provirus-derived beta3. The ability of this vector to target expression of Pl(A2)beta3 to megakaryocytes was first examined in cell lines. Immunoblot analysis with human anti-Pl(A2) alloserum detected synthesis of Pl(A2)beta3 in transduced promegakaryocytic cells; however, Pl(A2)beta3 protein was not detected in transduced epithelial cells. Human hematopoietic CD34+ cells were transduced with -889Pl(A2)beta3 virions and induced to differentiate with megakaryocyte growth and development factor. A hybrid alphaIIbbeta3 complex was formed in progeny megakaryocytes where provirus-derived Pl(A2)beta3 was detected associated with endogenous alphaIIb subunit. Another alphaIIb promoter-driven MuLV vector (-889nlacZ) encoding Escherichia coli beta-galactosidase was used to demonstrate that transgene expression was selectively targeted to the megakaryocyte progeny of transduced CD34+ cells. These studies demonstrate the feasibility of using alphaIIb promoter-driven MuLV vectors for gene transfer of hematopoietic CD34+ cells to target transgene expression in developing megakaryocytes and platelets and indicate potential applications toward human gene therapy for platelet disorders

Rap1b is critical for glycoprotein VI-mediated but not ADP receptor-mediated α 2 β 1 activation

The platelet α2β1 integrin functions as both an adhesion and signaling receptor upon exposure to collagen. Recent studies have indicated that α2β1 function can be activated via inside-out signaling, similar to the prototypical platelet integrin αIIbβ3. However, signaling molecules that regulate α2β1 activation in platelets are not well defined. A strong candidate molecule is the small GTPase Rap1b, the dominant platelet isoform of Rap1, which regulates αIIbβ3 activation

Search for the Rare Decay K_{L}\to\pi^{0}\pi^{0}\gamma

The KTeV E799 experiment has conducted a search for the rare decay $K_{L}\to\pi^{0}\pi^{0}\gamma$ via the topology $K_{L}\to\pi^{0}\pi^{0}_D\gamma$ (where $\pi^0_D\to\gamma e^+e^-$). Due to Bose statistics of the $\pi^0$ pair and the real nature of the photon, the $K_{L}\to\pi^{0}\pi^{0}\gamma$ decay is restricted to proceed at lowest order by the CP conserving direct emission (DE) of an E2 electric quadrupole photon. The rate of this decay is interesting theoretically since chiral perturbation theory predicts that this process vanishes at level $O(p^4)$. Therefore, this mode probes chiral perturbation theory at $O(p^6)$. In this paper we report a determination of an upper limit of $2.43\times 10^{-7}$ (90% CL) for $K_{L}\to\pi^{0}\pi^{0}\gamma$. This is approximately a factor of 20 lower than previous results.Comment: six pages and six figures in the submission. Reformatted for Physics Review

Search for the Rare Decays KL->pi0pi0mu+mu- and KL->pi0pi0X0->pi0pi0mu+mu-

The KTeV E799 experiment has conducted a search for the rare decays KL->pi0pi0mu+mu- and KL->pi0pi0X0->pi0pi0mu+mu-, where the X0 is a possible new neutral boson that was reported by the HyperCP experiment with a mass of (214.3 pm 0.5) MeV/c^{2}. We find no evidence for either decay. We obtain upper limits of Br(KL->pi0pi0X0->pi0pi0mu+mu-) pi0pi0mu+mu-) < 9.2 x 10^{-11} at the 90% confidence level. This result rules out the pseudoscalar X0 as an explanation of the HyperCP result under the scenario that the \bar{d}sX0 coupling is completely real

Determination of the Parity of the Neutral Pion via the Four-Electron Decay

We present a new determination of the parity of the neutral pion via the double Dalitz decay pi^0 -> e+ e- e+ e-. Our sample, which consists of 30511 candidate decays, was collected from K_L -> pi0 pi0 pi0 decays in flight at the KTeV-E799 experiment at Fermi National Accelerator Laboratory. We confirm the negative pi^0 parity, and place a limit on scalar contributions to the pi^0 -> e+ e- e+ e- decay amplitude of less than 3.3% assuming CPT conservation. The pi^0 gamma* gamma* form factor is well described by a momentum-dependent model with a slope parameter fit to the final state phase space distribution. Additionally, we have measured the branching ratio of this mode to be B(pi^0 -> e+ e- e+ e-) = (3.26 +- 0.18) x 10^(-5).Comment: 5 pages, 4 figures. Typographical error in radiative branching ratio (Eq. 6) correcte

Dispersive analysis of K_{L mu3} and K_{L e3} scalar and vector form factors using KTeV data

Using the published KTeV samples of K_L --> pi^{\pm} e^{\mp} nu and K_L --> pi^{\pm} mu^{\mp} nu decays [1], we perform a reanalysis of the scalar and vector form factors based on the dispersive parameterization [2,3]. We obtain phase space integrals I^e_K = 0.15446 \pm 0.00025 and I^{mu}_K = 0.10219 \pm 0.00025. For the scalar form factor parameterization, the only free parameter is the normalized form factor value at the Callan-Treiman point (C); our best fit results in ln C = 0.1915 \pm 0.0122. We also study the sensitivity of C to different parametrizations of the vector form factor. The results for the phase space integrals and C are then used to make tests of the Standard Model. Finally, we compare our results with lattice QCD calculations of F_K/F_pi and f_+(0).Comment: 9 pages, 3 figures, to be published in PR

The MAJORANA DEMONSTRATOR: A Search for Neutrinoless Double-beta Decay of Germanium-76

The {\sc Majorana} collaboration is searching for neutrinoless double beta decay using $^{76}$Ge, which has been shown to have a number of advantages in terms of sensitivities and backgrounds. The observation of neutrinoless double-beta decay would show that lepton number is violated and that neutrinos are Majorana particles and would simultaneously provide information on neutrino mass. Attaining sensitivities for neutrino masses in the inverted hierarchy region, $15 - 50$ meV, will require large, tonne-scale detectors with extremely low backgrounds, at the level of $\sim$1 count/t-y or lower in the region of the signal. The {\sc Majorana} collaboration, with funding support from DOE Office of Nuclear Physics and NSF Particle Astrophysics, is constructing the {\sc Demonstrator}, an array consisting of 40 kg of p-type point-contact high-purity germanium (HPGe) detectors, of which $\sim$30 kg will be enriched to 87% in $^{76}$Ge. The {\sc Demonstrator} is being constructed in a clean room laboratory facility at the 4850' level (4300 m.w.e.) of the Sanford Underground Research Facility (SURF) in Lead, SD. It utilizes a compact graded shield approach with the inner portion consisting of ultra-clean Cu that is being electroformed and machined underground. The primary aim of the {\sc Demonstrator} is to show the feasibility of a future tonne-scale measurement in terms of backgrounds and scalability.Comment: Proceedings for the MEDEX 2013 Conferenc