Esrrb is a prominent target of Nanog that substitutes for Nanog function in ES cell self-renewal, reprogramming and germline development

Embryonic stem (ES) cell pluripotency is sustained by a network of transcription factors centred on Oct4, Sox2 and Nanog. Whilst Oct4 and Sox2 expression is relatively uniform, ES cells fluctuate between states of high Nanog expression possessing high self-renewal efficiency, and low Nanog expression exhibiting increased differentiation propensity. Moreover, modulation in the level of Nanog expression determines the efficiency of ES cell self-renewal. To identify genes regulated by Nanog, genome-wide transcriptional profiling was performed on ES cells expressing different Nanog levels and Nanog-null ES cells expressing a Nanog-ERT2 fusion protein in which nuclear Nanog activity can be regulated by tamoxifen. Surprisingly, only a minor fraction of the genes to which Nanog binds showed significant changes in response to Nanog induction. Prominent amongst Nanog-responsive genes is Estrogen-related receptor b (Esrrb). Nanog binds directly to Esrrb, enhances binding and pause-release of RNAPolII from the Esrrb promoter and stimulates Esrrb transcription. Consistent with these findings, elevation of Nanog produces a cell population that expresses uniformly high Esrrb levels. Moreover, double fluorescent reporter lines show that Esrrb and Nanog levels are strongly correlated in individual cells. Loss of Nanog is required for downregulation of Esrrb, which coincides with commitment to differentiate. Esrrb overexpression results in LIF independent self-renewal, and blocks neural differentiation, even in the absence of Nanog. Cell fusion experiments between ES and neural stem (NS) cells show that elevated Esrrb levels allow the reprogramming of the NS cell genome in the absence of Nanog. Esrrb can rescue stalled reprogramming during the derivation of Nanog-/- induced pluripotent stem (iPS) cells. Moreover, targeted knock-in of Esrrb at the Nanog locus rescues the ability of Nanog null ES cells to maintain germ cell development beyond E12. Finally, Esrrb deletion abolishes the defining ability of Nanog to confer LIF-independent selfrenewal to ES cells. Together these data identify Esrrb as a critical downstream mediator of Nanog function

Black hole singularities in the framework of gauge/string duality

Thesis (Ph. D.)--Massachusetts Institute of Technology, Dept. of Physics, 2007.Includes bibliographical references (p. 201-209).In this dissertation black hole singularities are studied using the AdS/CFT correspondence. These singularities show up in the CFT in the behavior of finite-temperature correlation functions. A direct relation is established between space-like geodesics in the bulk and momentum space Wightman functions of CFT operators of large dimensions. This allows to probe the regions inside the horizon and near the singularity using the CFT. Information about the black hole singularity is encoded in the exponential falloff of finite-temperature correlators at large imaginary frequency. We also find a UV/UV connection that governs physics inside the horizon. For the case the bulk theory lives in 5 dimensions the dual theory is an SU(N) Yang-Mills theory on a sphere, a bounded many-body system. The signatures of the singularity we found are only present as N -+ oo. To elucidate the emergence of the singularity in the gauge theory we further study the large N limit. We argue that in the high temperature phase the theory is intrinsically non-perturbative in the large N limit. At any nonzero value of the 't Hooft coupling A, an exponentially large (in N2) number of free theory states of wide energy range (or order N) mix under the interaction. As a result the planar perturbation theory breaks down. We argue that an arrow of time emerges in the gauge theory and the dual string configuration should be interpreted as a stringy black hole.by Guido Nicola Innocenzo Festuccia.Ph.D

EKSTENSIFIKASI DAN INTENSIFIKASI PEMUNGUTAN PAJAK BUMI DAN BANGUNAN PERDESAAN DAN PERKOTAAN (PBB-P2) OLEH PEMERINTAH DAERAH KABUPATEN BOYOLALI

ABSTRAK EKSTENSIFIKASI DAN INTENSIFIKASI PEMUNGUTAN PAJAK BUMI DAN BANGUNAN PERDESAAN DAN PERKOTAAN (PBB-P2) OLEH PEMERINTAH DAERAH KABUPATEN BOYOLALI FALSONIA MAHARANI NIM F3413030 Tujuan dari penulisan Tugas Akhir ini adalah untuk mengetahui tingkat efektivitas ekstensifikasi dan intensifikasi PBB-P2 setelah Pemerintah Daerah Kabupaten Boyolali diberikan kewenangan untuk mengelola PBB-P2. Teknik pengumpulan data yang digunakan adalah mengumpulkan data primer dan data sekunder. Data primer diperoleh dari penelitian selama Praktek Kerja Lapangan di DPPKAD dan wawancara dengan beberapa staf bidang PBB-P2 serta dilengkapi dengan data sekunder yaitu dokumen dari DPPKAD dan dari buku. Hasil dari penelitian ini adalah jumlah Nomor Objek Pajak (NOP) dan realisasi terhadap tunggakan yang diperoleh dari KPP selalu bertambah setiap tahun, penerimaan PBB-P2 mencapai lebih dari 92% dari target yang ditetapkan serta jumlah tunggakan terhadap jumlah PBB-P2 yang terera dalam Surat Pemberitahuan Pajak Terutang (SPPT) PBB-P2 tidak mencapai 24%. Kesimpulan dari penelitian ini, upaya ekstensifikasi dan intensifikasi PBB-P2 di Kabupaten Boyolali dinilai efektif. Penulis memberikan saran kepada DPPKAD untuk melakukan pendataan ulang, penyuluhan terkait PBB-P2 secara rutin serta menambah jumlah SDM agar pelaksanaan ekstensifikasi dan intensifikasi PBB-P2 berjalan lebih optimal. Kata kunci: Ekstensifikasi Pajak, Intensifikasi Pajak, PBB-P

Esrrb Complementation Rescues Development of Nanog-Null Germ Cells.

The transcription factors (TFs) Nanog and Esrrb play important roles in embryonic stem cells (ESCs) and during primordial germ-cell (PGC) development. Esrrb is a positively regulated direct target of NANOG in ESCs that can substitute qualitatively for Nanog function in ESCs. Whether this functional substitution extends to the germline is unknown. Here, we show that germline deletion of Nanog reduces PGC numbers 5-fold at midgestation. Despite this quantitative depletion, Nanog-null PGCs can complete germline development in contrast to previous findings. PGC-like cell (PGCLC) differentiation of Nanog-null ESCs is also impaired, with Nanog-null PGCLCs showing decreased proliferation and increased apoptosis. However, induced expression of Esrrb restores PGCLC numbers as efficiently as Nanog. These effects are recapitulated in vivo: knockin of Esrrb to Nanog restores PGC numbers to wild-type levels and results in fertile adult mice. These findings demonstrate that Esrrb can replace Nanog function in germ cells

Reduced Oct4 expression directs a robust pluripotent state with distinct signaling activity and increased enhancer occupancy by Oct4 and Nanog

10.1016/j.stem.2013.04.023Cell Stem Cell125531-54

Multicenter Experience Using Total Lymphoid Irradiation and Antithymocyte Globulin as Conditioning for Allografting in Hematological Malignancies

A non myeloablative conditioning with total lymphoid irradiation (TLI) and antithymocyte globulin (ATG) was shown to protect against graft-versus-host disease (GVHD). To evaluate the effects of TLI-ATG in a multicenter study, 45 heavily pretreated patients, median age 51, with lymphoid (n = 38) and myeloid (n = 7) malignancies were enrolled at 9 centers. Twenty-eight patients (62%) received at least 3 lines of treatment before allografting, and 13 (29%) had refractory/relapsed disease at the time of transplantation. Peripheral blood hematopoietic cells were from HLA identical sibling (n = 30), HLA-matched (n = 9), or 1 antigen HLA-mismatched (n = 6) unrelated donors. A cumulative TLI dose of 8 Gy was administered from day −11 through −1 with ATG at the dose of 1.5 mg/kg/day (from day −11 through −7). GVHD prophylaxis consisted of cyclosporine and mycophenolate mofetil. Donor engraftment was reached in 95% of patients. Grade II to IV acute GVHD (aGVHD) developed in 6 patients (13.3%), and in 2 of these patients, it developed beyond day 100. Incidence of chronic GVHD (cGVHD) was 35.8%. One-year nonrelapse mortality was 9.1%. After a median follow-up of 28 months (range, 3-57 months) from transplantation, median overall survival was not reached, whereas median event-free survival was 20 months. This multicenter experience confirms that TLI-ATG protects against GVHD and maintains graft-vs-tumor effects