Screening the p53 status of human cell lines using a yeast functional assay

We have screened the p53 status of 156 human cell lines, including 142 tumor cell lines from 27 different tumor types and 14 cell lines from normal tissues by using functional analysis of separated alleles in yeast. This assay enables us to score wild-type p53 expression on the basis of the ability of expressed p53 to transactivate the reporter gene HIS3 via the p53-responsive GAL1 promotor in Saccharomyces cerevisiae. Of 142 tumor cell lines, at least 104 lines (73.2%) were found to express the mutated p53 gene: 94 lines (66.2%) were mutated in both alleles, three lines (2.1%) were heterozygous, and no p53 cDNA was amplified from seven lines (4.9%). Of the 14 cell lines originating from normal tissues, all the transformed or immortalized cell lines expressed mutant p53 only. Yeast cells expressing mutant p53 derived from 94 cell lines were analyzed for temperature-sensitive growth. p53 cDNA from eight cell lines showed p53-dependent temperature-sensitive growth, growing at 30 degrees C but not at 37 degrees C. Four temperature-sensitive p53 mutations were isolated: CAT-->CGT at codon 214 (H214R), TAC-->TGC at codon 234 (Y234C), GTG-->ATG at codon 272 (V272M), and GAG-->AAG (E285K). Functionally wild-type p53 was detected in 38 tumor cell lines (26.8%) and all of the diploid fibroblasts at early and late population doubling levels. These results strongly support the previous findings that p53 inactivation is one of the most frequent genetic events that occurs during carcinogenesis and immortalization.     

Molecular mechanism of regulation of pentylenetetrazol-induced calcium entry by 3'-untranslated region of a seizure-related cDNA, PTZ-17, in Xenopus oocytes

To determine the molecular mechanism of regulation of pentylenetetrazol (PTZ)-induced calcium entry by the seizure-related gene, PTZ-17, the role of the 3'-untranslated region (3'UTR) and also interaction between 3'UTR and intracellular factors were investigated. PTZ-induced calcium inward current in Xenopus oocytes injected with PTZ-17 RNA varied in magnitude among strains of mice: RNA derived from the DBA/2 mouse, which has a high susceptibility to convulsions, showed the largest current and that from the BALB/c mouse with a low susceptibility to convulsions showed no PTZ response. The sequence of 3'UTR showed alterations among mouse strains: 3'UTR of BALB/c showed a sequence alteration from T to G and that of DBA/2 showed a GTG insertion compared with that of B6. The 3'UTR also regulated the translation of chloramphenicol acetyltransferase (CAT) RNA depending on its sequence. A particular region within the 3'UTR demonstrated interaction with 60- and 47-kDa proteins. Sequence alterations in this region corresponded to disappearance or increase in PTZ-induced calcium entry. These findings suggest that a particular region within 3'UTR of the seizure-related gene, PTZ-17, is involved in PTZ-induced calcium entry via interaction between mRNA and specific RNA-binding proteins.     

Effects of the doping profile on current characteristics in BSITs

Proposed are two doping profiles of the conducting channel in bipolar-mode static induction transistors (BSITs) that improve the current handling capability. By using the results obtained from two-dimensional device simulations, it is shown that BSITs with the proposed profiles exhibit higher drain current density and DC current gain than conventional BSITs. Also discussed is the tradeoff between the current and voltage capabilities     

Design and synthesis of potent antitumor 5,4'-diaminoflavone derivatives based on metabolic considerations

Recently, we reported that 5,4'-diaminoflavone (1) exhibits potent and specific growth-inhibitory activity against the estrogen receptor (ER)-positive human breast cancer cell line MCF-7. However, when compound 1 was incubated with S-9 mix, its metabolites were observed. Moreover, addition of S-9 mix to the medium caused the drastic decrease in activity of compound 1. Since the 6-, 8-, and 3'-positions were considered to be metabolized oxidatively in vivo from MO calculations, a series of 5,4'-diaminoflavone derivatives substituted at such putative metabolic positions with various functional groups were synthesized aiming at the metabolically stable derivatives. Among them, 5,4'-diamino-6,8,3'-trifluoroflavone (14d) exhibited strong growth-inhibitory activity against MCF-7 cells even in the presence of S-9 mix. Moreover, orally administered compound 14d completely suppressed the growth of MCF-7 inoculated into nude mice, and the effect was more potent than that of compound 1. In addition to ER-positive breast cancer cells, compound 14d exhibited growth-inhibitory activity against a panel of human cancer cell lines including a part of ER-negative breast, endometrial, ovarian, and liver cancers. From these results, fluorine introduction to the putative metabolic positions of compound 1 was elucidated to be effective in the enhancement of the in vivo antitumor activity, probably due to the block of the metabolic deactivation.     

Magnetic Field Measurement System in Superconducting Combined Function Magnets for the J-PARC Neutrino Beam Line

Magnetic field measurements have been performed for the first full-scale magnet system assembled with the cryostat for the J-PARC proton beam line of neutrino experiment. In the measurement system, the probe position with respect to magnet central axis is measured by a Helium Neon laser and a position sensitive detector, PSD, in order to obtain an exact dipole field strength. Errors associated with the PSD misalignment and influences on the PSD signals by the magnetic field were evaluated. The measured dipole components approached to the design values by compensating those with the probe position. The latest beam simulation indicated that the measured values of magnetic field were good enough for the primary proton beam transport.     

A low-impedance open-bitline array for multigigabit DRAM

The noise-generating mechanisms inherent in the open-bitline DRAM array using the 6F² (F: feature size) memory cells and techniques for reducing the noise are described. The sources of differential noise coupled to the paired bitlines laid out in two arrays are the p-well, cell plate, and the group of nonselected wordlines. It was found, by simulation and by experiment with a 0.13-μm 256-Mb test chip, that the level of noise is dramatically reduced by using a low-impedance array with careful layout featuring low-resistivity materials, tight bridging between pairs of adjacent arrays, and a small array, achieving a comparable level of noise to that seen in the twisted and folded-bitline array. On basis of these results, it turns out that the open-bitline array has a strong chance of revival in the multigigabit generation, as long as these noise reduction techniques are applied     

Oscillating thromboemboli within the extracranial internal carotid artery demonstrated by ultrasonography in patients with acute cardioembolic stroke

BACKGROUND: Gunshot wounds to the kidney have been managed traditionally by surgical exploration. Concerns over increased nephrectomy rates and unnecessary explorations have placed this method under scrutiny. Selective renal exploration based on solid clinical and radiographic criteria may be a safer alternative and deserves evaluation. METHODS: The medical records of 52 consecutive patients with renal gunshot wounds, who were managed between September 1994 and August 1995 by a protocol of selective exploration, were reviewed. Renal injuries were explored only if they involved the hilum or were accompanied by signs of continued bleeding. Main outcome measures were the numbers of kidneys lost, as well as the morbidity and mortality related to the management of the renal injury. RESULTS: Three patients died from associated injuries shortly after admission. Fifteen patients suffered complications but only two had complications directly associated with the renal injury. Thirty-two patients underwent renal exploration and 17 of them required nephrectomy for major renovascular or parenchymal trauma (grade IV and V). Renal exploration was successfully avoided in the remaining 20 patients. No kidneys were lost unnecessarily as a result of this policy. CONCLUSION: Mandatory exploration of all gunshot wounds to the kidney is not necessary. Injuries that produce stable peripheral haematomas do not require exploration.     

A single receptor encoded by vzg-1/lpA1/edg-2 couples to G proteins and mediates multiple cellular responses to lysophosphatidic acid

Extracellular lysophosphatidic acid (LPA) produces diverse cellular responses in many cell types. Recent reports of several molecularly distinct G protein-coupled receptors have raised the possibility that the responses to LPA stimulation could be mediated by the combination of several uni-functional receptors. To address this issue, we analyzed one receptor encoded by ventricular zone gene-1 (vzg-1) (also referred to as lpA1/edg-2) by using heterologous expression in a neuronal and nonneuronal cell line. VZG-1 expression was necessary and sufficient in mediating multiple effects of LPA: [3H]-LPA binding, G protein activation, stress fiber formation, neurite retraction, serum response element activation, and increased DNA synthesis. These results demonstrate that a single receptor, encoded by vzg-1, can activate multiple LPA-dependent responses in cells from distinct tissue lineages.