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  • Generation of CD34+ cells from CCR5-disrupted human embryonic and induced pluripotent stem cells. 21981760

    C-C chemokine receptor type 5 (CCR5) is a major co-receptor for the entry of human immunodeficiency virus type-1 (HIV-1) into target cells. Human hematopoietic stem cells (hHSCs) with naturally occurring CCR5 deletions (Δ32) or artificially disrupted CCR5 have shown potential for curing acquired immunodeficiency syndrome (AIDS). However, Δ32 donors are scarce, heterologous bone marrow transplantation is not exempt of risks, and genetic engineering of autologous hHSCs is not trivial. Here, we have disrupted the CCR5 locus of human embryonic stem cells (hESCs) and induced pluripotent stem cells (hiPSCs) using specific zinc finger nucleases (ZFNs) combined with homologous recombination. The modified hESCs and hiPSCs retained pluripotent characteristics and could be differentiated in vitro into CD34(+) cells that formed all types of hematopoietic colonies. Our results suggest the potential of using patient-specific hHSCs derived from ZFN-modified hiPSCs for treating AIDS.
    Document Type:
    Reference
    Product Catalog Number:
    MAB4360
    Product Catalog Name:
    Anti-TRA-1-60 Antibody, clone TRA-1-60

  • Radioimmunoassay of a 26,000-dalton plasma insulin-like growth factor-binding protein: control by nutritional variables. 2461386

    Insulin-like growth factor I (IGF-I) is a peptide growth factor that circulates bound to carrier proteins. One form of carrier protein (mol wt, approximately 26K) is not believed to be GH dependent, is relatively unsaturated, and modulates the cellular response to IGF-I. This study was undertaken to determine the variables that control the plasma concentration of this protein, which was measured using a specific RIA. The mean plasma 26K IGF-binding protein (IGF-BP) concentration in 15 normal fasting subjects at 0800 h was 9.4 +/- 4.4 (+/- SD) micrograms/L. The mean value in GH-deficient patients was increased to 19.5 +/- 10.1 micrograms/L (n = 60; P less than 0.05), and it was 7.3 +/- 4.3 micrograms/L in patients with acromegaly (n = 31). The GH dependency of these changes is further supported by the observation that subjects who received GH injections had a 51% reduction in their fasting values. Nutritional intake appeared to be a more important controlling variable than GH. During an overnight fast plasma 26K IGF-BP values increased approximately 4-fold in 6 normal subjects. After 2 days of fasting, the mean value in 7 obese subjects rose progressively from 6.5 +/- 2.3 to 11.7 +/- 5.4 micrograms/L (P less than 0.001), and it increased further to 19.2 +/- 5.9 micrograms/L by day 4 of fasting; after 2 days of refeeding it returned to the prefasting level of 6.8 +/- 1.9 micrograms/L. Likewise, ingestion of a standard test meal resulted in a significant decrease in mean plasma 26K IGF-BP from a fasting value of 8.4 +/- 2.9 to 5.6 +/- 2.8 micrograms/L 4 h postprandially (P less than 0.05). In summary, the plasma concentrations of the 26K IGF-I-BP fluctuate widely in response to dietary manipulation, whereas GH status appears to be a secondary controlling variable.
    Document Type:
    Reference
    Product Catalog Number:
    Multiple
    Product Catalog Name:
    Multiple

  • Calorie restriction increases the ratio of phosphatidylinositol 3-kinase catalytic to regulatory subunits in rat skeletal muscle. 15613677

    Calorie restriction [CR; 60% of ad libitum (AL) intake] improves insulin-stimulated glucose transport, concomitant with enhanced phosphorylation of Akt. The mechanism(s) for the CR-induced increase in Akt phosphorylation of insulin-stimulated muscle is unknown. The purpose of this study was to determine whether CR increased the ratio of catalytic to regulatory subunits favoring enhanced phosphatidylinositol (PI) 3-kinase signaling, which may contribute to increases in Akt phosphorylation and glucose transport in insulin-stimulated muscles. We measured the PI 3-kinase regulatory (p85alpha/beta, p50alpha, and p55alpha) and catalytic (p110) subunits abundance in skeletal muscle from male F344B/N rats after 8 wk of AL or CR treatment. In CR compared with AL muscles, regulatory isoforms, p50alpha and p55alpha abundance were approximately 40% lower (P less than 0.01) with unchanged p85alpha/beta levels. There was no diet-related change in catalytic subunit abundance. Despite lower IRS-1 levels ( approximately 35%) for CR vs. AL, IRS-1-p110 association in insulin-stimulated muscles was significantly (P less than 0.05) enhanced by approximately 50%. Downstream of PI 3-kinase, CR compared with AL significantly enhanced Akt serine phosphorylation by 1.5-fold higher (P = 0.01) and 3-O-methylglucose transport by approximately 20% in muscles incubated with insulin. The increased ratio of PI 3-kinase catalytic to regulatory subunits favors enhanced insulin signaling, which likely contributes to greater Akt phosphorylation and improved insulin sensitivity associated with CR in skeletal muscle.
    Document Type:
    Reference
    Product Catalog Number:
    Multiple
    Product Catalog Name:
    Multiple
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