PURIFICATION AND CHARACTERIZATION OF A KINASE SPECIFIC FOR THE SERINE-RICH AND ARGININE-RICH PRE-MESSENGER-RNA SPLICING FACTORS

Members of the SR family of pre-mRNA splicing factors are phosphoproteins that share a phosphoepitope specifically recognized by monoclonal antibody (mAb) 104. Recent studies have indicated that phosphorylation may regulate the activity and the intracellular localization of these splicing factors. Here, we report the purification and kinetic properties of SR protein kinase 1 (SRPK1), a kinase specific for SR family members. We demonstrate that the kinase specifically recognizes the SR domain, which contains serine/arginine repeats. Previous studies have shown that dephosphorylated SR proteins did not react with mAb 104 and migrated faster in SDS gels than SR proteins from mammalian cells. We show that SRPK1 restores both mobility and mAB 104 reactivity to a SR protein SF2/ASF (splicing factor 2/alternative splicing factor) produced in bacteria, suggesting that SRPK1 is responsible for the generation of the mAb 104-specific phosphoepitope in vivo. Finally, we have correlated the effects of mutagenesis in the SR domain of SF2/ASF on splicing with those on phosphorylation of the protein by SRPK1, suggesting that phosphorylation of SR proteins is required for splicing.

Electron irradiation-induced defects in InP pre-annealed at high temperature

Electron irradiation-induced deep level defects have been studied in InP which has undergone high-temperature annealing in phosphorus and iron phosphide ambients, respectively. In contrast to a high concentration of irradiation-induced defects in as-grown and phosphorus ambient annealed InP, InP pre-annealed in iron phosphide ambient has a very low concentration of defects. The phenomenon has been explained in terms of a faster recombination of radiation-induced defects in the annealed InP. The radiation-induced defects in the annealed InP have been compared and studied. (c) 2006 Elsevier Ltd. All rights reserved.

Silicon nanowires grown on a pre-annealed Si substrate

Polycrystalline Si nanowires (poly SiNWS) were successfully synthesized by plasma-enhanced chemical vapor deposition (PECVD) at 440degreesC using silane as the Si source and Au as the catalyst. The diameters of Si nanowires range from 15 to 100nm. The growth process indicates that to fabricate SiNWS by PECVD, pre-annealing at high temperature is necessary. A few interesting nanowires with Au nanoclusters uniformly distributed in the body of the wire were also produced by this technique. (C) 2002 Elsevier Science B.V. All rights reserved.

The effects of pre-irradiation on the formation of Si1-xCx alloys

Carbon ions were implanted into crystal Si to a concentration of (0.6-1.5)at% at room temperature. Some samples were pre-irradiated with S-29(i)+ ions, while others were not pre-irradiated. Then the two kinds of samples were implanted with C-12(+) ions simultaneously, and Si1-xCx alloys were grown by solid phase epitaxy with high-temperature annealing. The effects of preirradiation on the formation of Si1-xCx alloys were studied. If the dose of implanted C ion was less than that for amorphizing Si crystals, the implanted C atoms would like to combine with defects produced during implantation, and then it was difficult for Si1-xCx alloys to form after annealine, at 950 degreesC. Pre-irradiation was advantageous for Si1-xCx alloy formation. With the increase of C ion dose, the damage produced by C ions increased. Pre-irradiation was unfavorable for Si1-xCx, alloy formation. If the implanted C concentration was higher than that for solid phase epitaxy solution, only part of the implanted C atoms form Si1-xCx alloys and the effects of pre-irradiation could be neglected. As the annealing temperature was increased to 1050 degreesC, Si1-xCx alloys in both pre-irradiated and unpreirradiated samples of low C concentration remained, whereas most part of Si1-xCx alloys in samples with high C concentration vanished.

Direct Wafer Bonding Technology employing vacuum-cavity pre-bonding

A novel low temperature direct wafer bonding technology employing vacuum-cavity pre-bonding is proposed and applied in bonding of InGaAs/Si couple wafers under 300 degrees C and InP/GaAs couple wafers under 350 degrees C. Aligning accuracy of 0.5 mu m is achieved. During wafer bonding process the pressure on the couple wafers is 10MPa. The interface energy is sufficiently high to allow thinning of the wafers down from 350um to about 100um. And the tensile strength test indicates the bonding energy of bonded samples is about equal to the bonded samples at 550 degrees C.

Electron irradiation-induced defects in InP pre-annealed at high temperature

Electron irradiation-induced deep level defects have been studied in InP which has undergone high-temperature annealing in phosphorus and iron phosphide ambients, respectively. In contrast to a high concentration of irradiation-induced defects in as-grown and phosphorus ambient annealed InP, InP pre-annealed in iron phosphide ambient has a very low concentration of defects. The phenomenon has been explained in terms of a faster recombination of radiation-induced defects in the annealed InP. The radiation-induced defects in the annealed InP have been compared and studied. (c) 2006 Elsevier Ltd. All rights reserved.

Alleviation of pre-exposure of mouse brain with low-dose C-12(6+) ion or Co-60 gamma-ray on male reproductive endocrine damages induced by subsequent high-dose irradiation

Irradiation has been widely reported to damage organisms by attacking on proteins, nucleic acid and lipids in cells. However, radiation hormesis after low-dose irradiation has become the focus of research in radiobiology in recent years. To investigate the effects of pre-exposure of mouse brain with low-dose C-12(6+) ion or Co-60 gamma (gamma)-ray on male reproductive endocrine capacity induced by subsequent high-dose irradiation, the brains of the B6C3F(1) hybrid strain male mice were irradiated with 0.05 Gy of C-12(6+) ion or Co-60 gamma-ray as the pre-exposure dose, and were then irradiated with 2 Gy as challenging irradiation dose at 4 h after pre-exposure. Serum pituitary gonadotropin hormones, follicle-stimulating hormone (FSH) and luteinizing hormone (LH), testosterone, testis weight, sperm count and shape were measured on the 35th day after irradiation. The results showed that there was a significant reduction in the levels of serum FSH, LH, testosterone, testis weight and sperm count, and a significant increase in sperm abnormalities by irradiation of the mouse brain with 2 Gy of C-12(6+) ion or Co-60 gamma-ray. Moreover, the effects were more obvious in the group irradiated by C-12(6+) ion than in that irradiated by Co-60 gamma-ray. Pre-exposure with low-dose C-12(6+) ion or Co-60 gamma-ray significantly alleviated the harmful effects induced by a subsequent high-dose irradiation.

Induction of cytogenetic adaptive response in spermatogonia and spermatocytes by pre-exposure of mouse testis to low-dose C-12(6+) ions

To investigate the effects of pre-exposure of mouse testis to low-dose C-12(6+) ions on cytogenetics of spermatogonia and spermatocytes induced by subsequent high-dose irradiation. the testes of outbred Kun-Ming strain mice were irradiated with 0.05 Gy of C-12(6+) ions as the pre-exposure dose, and then irradiated with 2 Gy as challenging dose at 4 h after per-exposure. Poly(ADP-ribose) polymerase (PARPs) activity and PARP-1 protein expression were respectively measured by using the enzymatic and Western blot assays at 4 h after irradiation; chromosomal aberrations in spermatogonia and spermatocytes were analyzed by the air-drying method at 8 h after irradiation. The results showed that there was a significant increase in the frequency of chromosomal aberrations and significant reductions of PARP activity and PARP-1 expression level in the mouse testes irradiated with 2 Gy of C-12(6+) ions. However, pre-exposure of mouse testes to a low dose of C-12(6+) ions significantly increased PARPs activity and PARP-1 expression and alleviated the harmful effects induced by a subsequent high-dose irradiation. PARP activity inhibitor 3-aminobenzamide (3-AB) treatment blocked the effects of PARP-1 on cytogenetic adaptive response induced by low-dose C-12(6+) ion irradiation. The data suggest that pre-exposure of testes to a low dose of heavy ions can induce cytogenetic adaptive response to subsequent high-dose irradiation. The increase of PARP-1 protein induced by the low-dose ionizing irradiation may be involved in the mechanism of these observations. (C) 2008 Elsevier B.V. All rights reserved.

Pre-irradiation with low-dose C-12(6+) beam significantly enhances the efficacy of AdCMV-p53 gene therapy in human non-small lung cancer

The combination of ionizing radiation and gene therapy has been investigated. However, there are very few reports about the combination of heavy-ion irradiation and gene therapy. To determine if the pre-exposure to low-dose heavy ion beam enhances the suppression of AdCMV-p53 on non-small lung cancer (NSLC), the cells pre-irradiated or non-irradiated were infected with 20, 40 MOI of AdCMV-p53. Survival fraction and the relative biology effect (RBE) were determined by clonogenic assay. The results showed that the proportions of p53 positive cells in C-12(6+) beam induced AdCMV-p53 infected cells were more than 90%, which were significantly more than those in gamma-ray induced AdCMV-p53 infected cells. The pre-exposure to low-dose 12C6+ beam significantly prevented the G(0)/G(1) arrest and activated G(2)/M checkpoints. The pre-exposure to C-12(6+) beam significantly improved cell to apoptosis. RBEs for the C-12(6+)+ AdCMV-p53 infection groups were 30%-60%,20% -130% and 30%-70% more than those for the C-12(6+)_irradiated only, AdCMV-p53 infected only, and gamma-irradiation induced AdCMVp53 infected groups, respectively. The data suggested that the pre-exposure to low-dose C-12(6+) beam significantly promotes exogenous p53 expression in NSLC, and the suppression of AdCMV-p53 gene therapy on NSLC.