20 research outputs found

    Germline Competent Pluripotent Mouse Stem Cells Generated by Plasmid Vectors

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    <p>We developed nonintegrated methods to reprogram mouse embryonic fibroblast (MEF) cells into induced pluripotent stem cells (iPSCs) using pig pOct4, pSox2, and pc-Myc as well as human hKLF4, hAID, and hTDG that were carried by plasmid vectors. The 4F method employed pOct4, pSox2, pc-Myc, and hKLF4 to derive iPSC clones with naive embryonic stem cell (ESC)-like morphology. These 4F clones expressed endogenous mouse Nanog protein and could generate chimeras. In addition to the four conventional reprogramming factors used in the 4F method, hAID and hTDG were utilized in a 6F method to increase the conversion efficiency of reprogramming by approximately five-fold. One of the 6F plasmid derived iPSC (piPSC) clones was shown to be germline transmission competent.</p

    Insight into the Reactivity and Electronic Structure of Dinuclear Dinitrosyl Iron Complexes

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    A combination of N/S/Fe K-edge X-ray absorption spectroscopy (XAS), X-ray diffraction data, and density functional theory (DFT) calculations provides an efficient way to unambiguously delineate the electronic structures and bonding characters of Fe–S, N–O, and Fe–N bonds among the direduced-form Roussin’s red ester (RRE) [Fe<sub>2</sub>(μ-SPh)<sub>2</sub>(NO)<sub>4</sub>]<sup>2–</sup>(<b>1</b>) with {Fe­(NO)<sub>2</sub>}<sup>10</sup>-{Fe­(NO)<sub>2</sub>}<sup>10</sup> core, the reduced-form RRE [Fe<sub>2</sub>(μ-SPh)<sub>2</sub>(NO)<sub>4</sub>]<sup>−</sup>(<b>3</b>) with {Fe­(NO)<sub>2</sub>}<sup>9</sup>-{Fe­(NO)<sub>2</sub>}<sup>10</sup> core, and RRE [Fe<sub>2</sub>(μ-SPh)<sub>2</sub>(NO)<sub>4</sub>] (<b>4</b>) with {Fe­(NO)<sub>2</sub>}<sup>9</sup>-{Fe­(NO)<sub>2</sub>}<sup>9</sup> core. The major contributions of highest occupied molecular orbital (HOMO) 113α/β in complex <b>1</b> is related to the antibonding character between Fe­(d) and Fe­(d), Fe­(d), and S atoms, and bonding character between Fe­(d) and NO­(π*). The effective nuclear charge (<i><i>Z</i></i><sub>eff</sub>) of Fe site can be increased by removing electrons from HOMO to shorten the distances of Fe···Fe and Fe–S from <b>1</b> to <b>3</b> to <b>4</b> or, in contrast, to increase the Fe–N bond lengths from <b>1</b> to <b>3</b> to <b>4</b>. The higher IR ν<sub>NO</sub> stretching frequencies (1761, 1720 cm<sup>–1</sup> (<b>4</b>), 1680, 1665 cm<sup>–1</sup> (<b>3</b>), and 1646, 1611, 1603 cm<sup>–1</sup> (<b>1</b>)) associated with the higher transition energy of N<sub>1s</sub> →σ*­(NO) (412.6 eV (<b>4</b>), 412.3 eV (<b>3</b>), and 412.2 eV (<b>1</b>)) and the higher <i><i>Z</i></i><sub>eff</sub> of Fe derived from the transition energy of Fe<sub>1s</sub> → Fe<sub>3d</sub> (7113.8 eV (<b>4</b>), 7113.5 eV (<b>3</b>), and 7113.3 eV (<b>1</b>)) indicate that the N–O bond distances of these complexes are in the order of <b>1 > 3 > 4</b>. The N/S/Fe K-edge XAS spectra as well as DFT computations reveal the reduction of complex <b>4</b> yielding complex <b>3</b> occurs at Fe, S, and NO; in contrast, reduction mainly occurs at Fe site from complex <b>3</b> to complex <b>1</b>

    Insight into the Reactivity and Electronic Structure of Dinuclear Dinitrosyl Iron Complexes

    No full text
    A combination of N/S/Fe K-edge X-ray absorption spectroscopy (XAS), X-ray diffraction data, and density functional theory (DFT) calculations provides an efficient way to unambiguously delineate the electronic structures and bonding characters of Fe–S, N–O, and Fe–N bonds among the direduced-form Roussin’s red ester (RRE) [Fe<sub>2</sub>(μ-SPh)<sub>2</sub>(NO)<sub>4</sub>]<sup>2–</sup>(<b>1</b>) with {Fe­(NO)<sub>2</sub>}<sup>10</sup>-{Fe­(NO)<sub>2</sub>}<sup>10</sup> core, the reduced-form RRE [Fe<sub>2</sub>(μ-SPh)<sub>2</sub>(NO)<sub>4</sub>]<sup>−</sup>(<b>3</b>) with {Fe­(NO)<sub>2</sub>}<sup>9</sup>-{Fe­(NO)<sub>2</sub>}<sup>10</sup> core, and RRE [Fe<sub>2</sub>(μ-SPh)<sub>2</sub>(NO)<sub>4</sub>] (<b>4</b>) with {Fe­(NO)<sub>2</sub>}<sup>9</sup>-{Fe­(NO)<sub>2</sub>}<sup>9</sup> core. The major contributions of highest occupied molecular orbital (HOMO) 113α/β in complex <b>1</b> is related to the antibonding character between Fe­(d) and Fe­(d), Fe­(d), and S atoms, and bonding character between Fe­(d) and NO­(π*). The effective nuclear charge (<i><i>Z</i></i><sub>eff</sub>) of Fe site can be increased by removing electrons from HOMO to shorten the distances of Fe···Fe and Fe–S from <b>1</b> to <b>3</b> to <b>4</b> or, in contrast, to increase the Fe–N bond lengths from <b>1</b> to <b>3</b> to <b>4</b>. The higher IR ν<sub>NO</sub> stretching frequencies (1761, 1720 cm<sup>–1</sup> (<b>4</b>), 1680, 1665 cm<sup>–1</sup> (<b>3</b>), and 1646, 1611, 1603 cm<sup>–1</sup> (<b>1</b>)) associated with the higher transition energy of N<sub>1s</sub> →σ*­(NO) (412.6 eV (<b>4</b>), 412.3 eV (<b>3</b>), and 412.2 eV (<b>1</b>)) and the higher <i><i>Z</i></i><sub>eff</sub> of Fe derived from the transition energy of Fe<sub>1s</sub> → Fe<sub>3d</sub> (7113.8 eV (<b>4</b>), 7113.5 eV (<b>3</b>), and 7113.3 eV (<b>1</b>)) indicate that the N–O bond distances of these complexes are in the order of <b>1 > 3 > 4</b>. The N/S/Fe K-edge XAS spectra as well as DFT computations reveal the reduction of complex <b>4</b> yielding complex <b>3</b> occurs at Fe, S, and NO; in contrast, reduction mainly occurs at Fe site from complex <b>3</b> to complex <b>1</b>

    Alkaline phosphatase (AP) activity and karyotypes of ntES cells derived from aggregated cloned blastocysts.

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    <p>(a) A phase-contrast image of ntES cell colony with positive AP activity at passage 3. (b) Karyotyping by Giemsa staining of ntES cell line PES1 at passage 20 with 75% of normal karyotype, and (c) PES3 line at passage 28 with a normal karyotype ratio of 85%. Scale bar = 100 μm.</p

    Formation of ntES cell colonies derived from the blastocyst embryos of the 3× cloned embryos.

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    <p>(a) A 3× blastocyst embryo (day 7) and (b) embryonal outgrowths with clearly ICM cells (red arrow) after culture for 5–8 days. (c) Typical morphology of a putative porcine ntES cell colony after passage. Scale bar = 100 μm.</p

    Differentiation of ntES cells in embryoid bodies (EB) and RT-PCR detection of genes typical for three germ layers.

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    <p>(a) Light microscopic images of EB cultured for 10 days. (b) Expressions of the genes representing all three germ layers, GATA4 (endoderm), β-III tubulin (ectoderm), and BMP-4 (mesoderm) in EB derived from PES1 (A), PES3 (B), and the negative control is represented by undifferentiated PES3 cells (C). Scale bars = 100 μm.</p
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