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Daniel Fisher

Publications and source records attributed to Daniel Fisher.

7 recordsLinked to original sources
PubMed2006

Only Akt1 is required for proliferation, while Akt2 promotes cell cycle exit through p21 binding.

Protein kinase B (PKB/Akt) is an important modulator of insulin signaling, cell proliferation, and survival. Using small interfering RNA duplexes in nontransformed mammalian cells, we show that only Akt1 is essential for cell proliferation, while Akt2 promotes cell cycle exit. Silencing Akt1 resulted in decreased cyclin A levels and inhibition of S-phase entry, effects not seen with Akt2 knockdown and specifically rescued by microinjection of Akt1, not Akt2. In differentiating myoblasts, Akt2 knockout prevented myoblasts from exiting the cell cycle and showed sustained cyclin A expression. In contrast, overexpression of Akt2 reduced cyclin A and hindered cell cycle progression in M-G1 with increased nuclear p21. p21 is a major target in the differential effects of Akt isoforms, with endogenous Akt2 and not Akt1 binding p21 in the nucleus and increasing its level. Accordingly, Akt2 knockdown cells, and not Akt1 knockdown cells, showed reduced levels of p21. A specific Akt2/p21 interaction can be reproduced in vitro, and the Akt2 binding site on p21 is similar to that in cyclin A spanning T145 to T155, since (i) prior incubation with cyclin A prevents Akt2 binding, (ii) T145 phosphorylation on p21 by Akt1 prevents Akt2 binding, and (iii) binding Akt2 prevents phosphorylation of p21 by Akt1. These data show that specific interaction of the Akt2 isoform with p21 is key to its negative effect on normal cell cycle progression.

Animals↗
PubMed2005

Transfer of transdermally applied testosterone to clothing: a comparison of a testosterone patch versus a testosterone gel.

AIM: To assess the amount of testosterone transferred from the abdominal application sites of a transdermal testosterone patch and a transdermal testosterone gel to cotton T-shirts worn for 24 hours during each treatment. METHODS: During a crossover study comparing the pharmacokinetics of a testosterone patch versus a testosterone gel in 28 hypogonadal men, subjects wore fresh cotton T-shirts for 24 hours on the seventh and fourteenth days of each treatment and during a 24-hour baseline period. At the end of each evaluation, the abdominal section of the shirt was carefully cut out, extracted in alcohol, and analyzed for testosterone by a direct radioimmunoassay. The minimum quantifiable amount of testosterone was 0.03 microg per T-shirt sample. RESULTS: The median amounts of extracted testosterone were 0.44 microg for the baseline samples, 25.4 microg for the average of the two patch samples, and 6,762.7 microg for the average of the two gel samples (all comparisons P < 0.0001). Significant correlations were observed between the day 7 and day 14 data for both the patch (R = 0.4982; P < 0.01) and the gel (R = 0.8383; P < 0.0001). No significant correlations were found between the baseline, patch, or gel data, or between these and any demographic or pharmacokinetic parameters. A quantitative interpretation of the findings suggests that the baseline results are consistent with the transfer of testosterone in sweat; the patch results are probably due to the transfer of a small amount of residual testosterone left on the abdominal skin from the prior day's patches; and the gel results reflect the desquamation of stratum corneum containing a portion of the abdominally applied testosterone. CONCLUSIONS: There are large differences in the amount of testosterone that can be transferred from the application sites of transdermal patches and gels. The latter should be covered with clothing to minimize transfer through intimate skin-to-skin contact.

Abdomen↗
PubMed2004

A history of Combined-Integrated doctoral training in psychology.

The history of doctoral training in psychology is reviewed, with special emphasis on the development of training models for applied psychology. Among applied psychology doctoral programs, most label themselves as being one of the three currently recognized practice areas: clinical, counseling, and school psychology; however, since the 1970s, APA also has accredited "combined" programs. The history of the development of the combined model is traced, including information from the programs themselves about the manner in which each evolved. Recently, new interest in the combined category -or Combined-Integrated (C-I) as these programs wish to become known-has emerged, and an organization representing these programs has formed. The C-I model is described, and the potential that the model may serve as a template for the future of professional training in psychology is discussed.

Education, Graduate↗
PubMed2003

Permanent cell cycle exit in G2 phase after DNA damage in normal human fibroblasts.

Although the Cdk inhibitor p21(Waf1/Cip1), one of the transcriptional targets of p53, has been implicated in the maintenance of G(2) arrest after DNA damage, its function at this stage of the cell cycle is not really understood. Here, we show that the exposure of normal human fibroblasts (NHFs) to genotoxic agents provokes permanent cell cycle exit in G(2) phase, whereas mouse embryo fibroblasts and transformed human cells progress through mitosis and arrest in G(1) without intervening cytokinesis. p21(Waf1/Cip1) exerts a key role in driving this G(2) exit both by inhibiting cyclin B1-Cdk1 and cyclin A-Cdk1/2 complexes, which control G(2)/M progression, and by blocking the phosphorylation of pRb family proteins. NHFs with compromised pRb proteins could still efficiently arrest in G(2) but were unable to exit the cell cycle, resulting in cell death. Our experiments show that, when under continuous genotoxic stress, normal cells can reverse their commitment to mitotic progression due to passage through the restriction point and that mechanisms involving p21(Waf1/Cip1) and pocket proteins can induce exit in G(2) and G(1).

Bleomycin↗
PubMed2003

Vertebrate HoxB gene expression requires DNA replication.

To study the relationship between DNA replication and transcription in vivo, we investigated Hox gene activation in two vertebrate systems: the embryogenesis of Xenopus and the retinoic acid-induced differentiation of pluripotent mouse P19 cells. We show that the first cell cycles following the mid- blastula transition in Xenopus are necessary and sufficient for HoxB activation, whereas later cell cycles are necessary for the correct expression pattern. In P19 cells, HoxB expression requires proliferation, and the entire locus is activated within one cell cycle. Using synchronous cultures, we found that activation of HoxB genes is colinear within a single cell cycle, occurs during S phase and requires S phase. The HoxB locus replicates early, whereas replication is still required for maximal expression later in S phase. Thus, induction of HoxB genes occurs in a DNA replication-dependent manner and requires only one cell cycle. We propose that S-phase remodelling licenses the locus for transcriptional regulation.

Animals↗