Nude Mature Wendy ~REPACK~

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Effect of OPG overexpression by breast cancer cells on tumor growth. (A) Five week old female nude mice were injected with 0.5105 MDA-MB-231-TXSA-TGL-pRUF or p-OPG cells directly into the marrow cavity of their right tibia. Mice were imaged weekly using the Xenogen IVIS 100 bioluminescence imaging system. Representative whole body BLI images of a single animal from each group (n=10/cell line) during the course of the experiments are shown. All animals, in both groups, were humanely sacrificed on Day 28 for ethical reasons, due to high tumor load. (B) The line graph represents the average tumor signal over time measured as mean photon counts per second and it demonstrates that OPG overexpression has no effect on tumor growth when compared to p-RUF tumors. (C) OPG concentration in the blood serum of mice (n=10/cell line) collected at two different time points during the experiment as measured by ELISA, bars SEM.

While InsP3 can generate calcium oscillations in vivo, it can also produce higher ordered InsPs [26,53,54]. We monitored the production of InsP6 using both isotope labeling and mass measurement. The [3H]myo-inositol labeling of seedlings revealed a significant increase in [3H]-labeled InsP6 indicating that the increase in InsP3 had affected higher ordered InsP biosynthesis (Figure 6C). To assess total InsP6, we used isocratic ion chromatography as described in the Experimental Section. In a preliminary experiment, we did not detect differences in total InsP6 in seedlings (data not shown). Because it was difficult to obtain enough material to do InsP6 mass measurements on the seedlings, we analyzed seeds (Figure 7A). InsP6 is produced by two pathways: a lipid-mediated pathway resulting from the phosphorylation of lipid-generated Ins(1,4,5)P3 and a non-lipid-dependent pathway, which involves de novo synthesis and the sequential phosphorylation of myo-1L-inositol phosphate [53]. The non-lipid-dependent pathway is the dominant pathway in storage tissue [55] and as shown in Figure 7A, the seeds from the HsPIPK1α plants had 40% less total InsP6. Typically seeds with low InsP6 have high Pi [56,57] and this is what we found for the HsPIPK1α. The seed HOAc-soluble Pi in the HsPIPK1α lines was almost twice that of the controls (Figure 7B). In contrast, the seedling HOAc-soluble Pi was about 20% less than wild type (Figure 7C) and there was no significant difference in HOAc-soluble Pi in 2-month-old mature leaves (Figure 7D). These data suggest that down regulation of the non-lipid-dependent pathway was compensating for the increased flux through the PI pathway in seedlings and leaves, and that in seeds where the non-lipid pathway was dominant, down regulation led to a net decrease in InsP6. More extensive flux analyses of both the lipid- and non-lipid mediated pathway for InsP6 biosynthesis are needed in order to determine whether the non-lipid pathway is down regulated in the leaves of the HsPIPK1α plants.

During wound healing, stem cells provide functional mature cells to meet acute demands for tissue regeneration (1). Simultaneously, the tissue must maintain a pool of stem cells to sustain its future regeneration capability. However, how these requirements are balanced in response to injury is unknown. Here we demonstrate that after wounding or ultraviolet type B irradiation, melanocyte stem cells (McSCs) in the hair follicle (2) exit the stem cell niche before their initial cell division, potentially depleting the pool of these cells. We also found that McSCs migrate to the epidermis in a melanocortin 1 receptor (Mc1r)-dependent manner and differentiate into functional epidermal melanocytes, providing a pigmented protective barrier against ultraviolet irradiation over the damaged skin. These findings provide an example in which stem cell differentiation due to injury takes precedence over stem cell maintenance and show the potential for developing therapies for skin pigmentation disorders by manipulating McSCs. 041b061a72