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In this study, we analyzed the roles for AML1/RUNX1 in the regulation of the c-mpl promoter. Wild-type AML1 activated the c-mpl promoter through the proximal AML-binding site in luciferase assays using 293T and HeLa cells. In accord with this result, electrophoretic mobility shift assay and chromatin immunoprecipitation assays demonstrated that AML1 bound to this site. Next, we analyzed the function of AML1 using a mutant of AML1 lacking the C terminus (AML1dC), which was originally found in a patient with myelodysplastic syndromes. AML1dC dominant-negatively suppressed transcriptional activity of wild-type AML1. However, unexpectedly, AML1dC-transduced murine c-Kit(+)Sca1(+)Lineage(-) cells expressed c-mpl mRNA and c-Mpl protein more abundantly than mock-transduced cells, which led to the enhanced thrombopoietin-mediated proliferation. Moreover, when AML1dC was induced to express during the development of hematopoietic cells from embryonic stem (ES) cells, AML1dC augmented the c-Mpl expression on hematopoietic stem/progenitor cells. Furthermore, we found that early hematopoietic cells that derived from AML1(-/-) ES cells expressed c-Mpl more intensely than those that developed from wild-type ES cells. In contrast, AML1dC hardly affected c-Mpl expression and maturation of megakaryocytes. As for the mechanism of the different roles of AML1 in the regulation of the c-mpl promoter, we found that AML1 forms a complex with a transcription repressor mSin3A on the c-mpl promoter in hematopoietic stem/progenitor cells, although it forms a complex with a transcription activator p300 on the same promoter in megakaryocytic cells. Together, these data indicate that AML1 can regulate the c-mpl promoter both positively and negatively by changing the binding partner according to cell types.

Notch and HOXB4 have been reported to expand hematopoietic stem cells (HSCs) in vitro. However, their critical effector molecules remain undetermined. We found that the expression of c-myc, cyclin D2, cyclin D3, cyclin E, and E2F1 was induced or enhanced during Notch1- or HOXB4-induced self-renewal of murine HSCs. Since c-Myc can act as a primary regulator of G1/S transition, we examined whether c-Myc alone can induce self-renewal of HSCs. In culture with stem cell factor, FLT3 ligand, and IL-6, a 4-hydroxytamoxifen-inducible form of c-Myc (Myc/ERT) enabled murine Lin- Sca-1+ HSCs to proliferate with the surface phenotype compatible with HSCs for more than 28 days. c-Myc activated by 4-hydroxytamoxifen augmented telomerase activities and increased the number of CFU-Mix about 2-fold in colony assays. Also, in reconstitution assays, HSCs expanded by c-Myc could reconstitute hematopoiesis for more than 6 months. As for the mechanism of c-myc induction by Notch1, we found that activated forms of Notch1 (NotchIC) and its downstream effector recombination signal-binding protein-J kappa (RBP-VP16) can activate the c-myc promoter through the element between -195 bp and -161 bp by inducing the DNA-binding complex. Together, these results suggest that c-Myc can support self-renewal of HSCs as a downstream mediator of Notch and HOXB4.

Recently, Kiritani et al. proposed a new mechanism of plastic deformation without involving dislocations in tensile fracture of metal foils. The paper reports transmission electron microscopy (TEM) study of tensile fracture of Al containing hard precipitates (Si) that are considered to act as obstacles to dislocation motion. In sawtooth-shaped thin-foils formed at the fracture tip (‘sawtooth portion’), tensile strain was as high as 103, but only a few dislocations were pinned to precipitates. Instead, voids were formed at precipitate/matrix interface, elongated in the direction of tension, and broke up into several smaller voids, due to stress concentration around hard precipitates. The thicker area of the specimen (‘base portion’), where tensile strain was 30, did not contain voids but showed a dislocation cell structure. In tensile fracture of pre-thinned specimen, voids were formed in the sawtooth portion, despite the tensile strain also being 30. These results suggest that the sawtooth portion is formed by a new mechanism that does not involve dislocations.

Based on computer simulations, we examined a new mechanism of plastic deformation that has been proposed to operate in tensile fracture of metal foils. We constructed a Au crystal containing high concentration of vacancies and/or one subjected to large elastic tensile strain using embedded atom method (EAM) potential, and then calculated transmission electron microscopy (TEM) images of the crystal based on multislice method. Randomly distributed vacancies did not cause a large distortion in the crystal lattice, and did not affect the TEM image intensity appreciably unless the vacancy concentration exceeded several percent. Under a large elastic tensile strain of about 10%along 100, a periodic displacement of atoms whose amplitude was 10%of the atomic distance was induced, reducing the intensity of equal thickness fringe by about half. At around 15%tensile strain along 110, the crystal transformed from fcc to bcc structure, releasing the distortion of crystal lattice.

Copper base binary alloys have been irradiated with 1 MeV electrons using a high-voltage electron microscope in order to study solute-point defect interactions and their effects on defect structure development. This paper reports results on Cu-Pd and Cu-Pt, and compares them with previous results on Cu-Ni, -Si, -Ge, and -Sn. Pd and Pt have a similar volume size factor as Ge (about +30%), and they belong to the same group as Ni (an undersize solute) in the periodic table of elements. At lower temperatures, the addition of Pd and Pt was found to stabilize interstitial-type dislocation loops, but did not increase the loop number density as drastically as the addition of Si, Ge, or Sn, Addition of 2 at.% of Pd or Pt resulted in the formation of stacking fault tetrahedra (SFTs) stable up to higher temperatures, and also voids between 373 It and 523 It. 0.3 at.% of Pd or Pt, however, did not induce either stable SFTs or voids. In contrast, addition of 0.3 at.% Si, Ge, and Sn was found to stabilize SFTs. These results suggest that solute-point defect interactions are not characterized only by atomic volume size factor.

For self-interstitial atom (SIA) clusters in various concentrated alloys, one-dimensional (1D) migration is induced by electron irradiation around 300 K. But at elevated temperatures, the 1D migration frequency decreases to less than one-tenth of that around 300 K in iron-based bcc alloys. In this study, we examined mechanisms of 1D migration at elevated temperatures using in situ observation of SUS316L and its model alloys with high-voltage electron microscopy. First, for elevated temperatures, we examined the effects of annealing and short-term electron irradiation of SIA clusters on their subsequent 1D migration. In annealed SUS316L, 1D migration was suppressed and then recovered by prolonged irradiation at 300 K. In high-purity model alloy Fe-18Cr-13Ni, annealing or irradiation had no effect. Addition of carbon or oxygen to the model alloy suppressed 1D migration after annealing. Manganese and silicon did not suppress 1D migration after annealing but after short-term electron irradiation. The suppression was attributable to the pinning of SIA clusters by segregated solute elements, and the recovery was to the dissolution of the segregation by interatomic mixing under electron irradiation. Next, we examined 1D migration of SIA clusters in SUS316L under continuous electron irradiation at elevated temperatures. The 1D migration frequency at 673 K was proportional to the irradiation intensity. It was as high as half of that at 300 K. We proposed that 1D migration is controlled by the competition of two effects: induction of 1D migration by interatomic mixing and suppression by solute segregation.