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http://game-rapidshare.com/Martling-father-of-Lascody-from-London?Skardagra=170
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    Cheeky

    A post shared by D E M I • 19 • ?? (@demi.rawling) on

    Demi hit her lowest weight of 7st in 2015. She said: “It got to the point where I would look in the mirror and I was unrecognisable.

    “I was stepping on the scales and realising just how low the numbers were.”

    She put on half a stone and convinced herself she had recovered, but was still desperately thin.

    Anorexia stopped sociable Demi going out with friends, and soon she was only interacting with people through YouTube make-up tutorials.

    She said: “I felt like they were like friends, because I was so isolated.

    “It was the only time I wasn’t consumed by my eating disorder. I wanted to make videos and work as a make-up artist.

    “But, at 18, something just snapped and I realised that if I wanted the life I was envisioning, I couldn’t do it if I was sick, or if I had to spend all day in bed – too thin or depressed to do anything.

    “I felt like I was wasting my life.”

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    Mice, parasites, and cell culture

    129S-PARP1tm1Zqw/J (PARP1-/-) mice were crossed with C57BL/6 mice to generate PARP1-/- mice on 129S/BL6 genetic background. PARP1-/- mice were bred with WT mice to generate PARP1+/- mice. All breeding pairs were purchased from Jackson Laboratory (Bar Harbor ME), and a standard PCR was performed to confirm the genotype of WT (PARP1+/+), PARP1+/- and PARP1-/- mice (S1A Fig).

    T. cruzi (SylvioX10/4, ATCC 50823) was propagated by in vitro passage in C2C12 cells. The WT, PARP1+/- and PARP1-/- mice (all 129S/BL6 background, 6-weeks-old) were infected with T. cruzi (10,000 trypomastigotes/mouse, intraperitoneal). For some studies, C57BL/6 mice were infected as above, and then mice were given a treatment of 2-(dimethylamino)-N-(6-oxo-5,6-dihydrophenanthridin-2-yl)acetamide hydrochloride (PJ34, Sigma-Aldrich, St Louis MO). PJ34 is a cell-permeable, water-soluble, selective PARP1 inhibitor PJ34 (EC50 = 20 nM) and shown to be ~10,000 times more potent than the prototypical PARP inhibitor, 3-aminobenzamide [20,21]. PJ34 (12.5 mg/kg) was delivered intraperitoneally for three weeks (twice a week) beginning at 45 days’ post-infection (pi) when acute parasitemia was controlled. All mice were harvested at 150 days’ pi corresponding to chronic disease phase. Sera/plasma and tissue samples were stored at 4°C and -80°C, respectively.

    Human cardiomyocyte cells (AC16, cat#SCC109, EMD Millipore, Burlington MA) were cultured and maintained in Dulbecco’s modified Eagle’s medium (DMEM)/F-12 medium containing 12.5% fetal bovine serum (FBS). Human cervix epithelial cells (HeLa, ATCC, Manassas VA) were propagated in DMEM media supplemented with Earle's salts, 2 mM L-glutamine and 10% FBS. Cells were infected with T. cruzi (cell: parasite ratio: 1:5) for various studies.

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    “He first started off with chemotherapy to the trachea, and after a while he was starting to feel fine,” Diane said. “He went home after three days of chemo and had his first seizure on the porch, so they put him right back into the hospital.”

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