A novel technique of effectively producing highly charged ions in laser produced plasma is demonstrated. Time-resolved and integrated soft X-ray spectra showed that ionization of Al atoms is greatly promoted up to Al10+ through the use of a pulse train laser even at very low peak power (1011 W/cm2). The X-ray conversion efficiency increased with increasing number of pulses and with decreasing interpulse time. The interpretation of our experimental results is presented.
Several methods have been reported for the semi-quantitative detection of small amounts of a particular mRNA in tissues through specific amplification of the extracted mRNA by reverse transcriptase polymerase chain reaction (RT-PCR), usually followed by agarose gel electrophoretic separation and ethidium bromide staining for fluorescence detection. [1][2][3][4] However, most of the methods are not suitable for accurate quantification owing to lack of correction for incomplete recovery of mRNA during extraction from the tissue or subsequent PCR procedures. It is also difficult to avoid PCR amplification into a non-linear region because of insufficient detection sensitivity. To overcome these problems, competitive RT-PCR with an internal standard has been shown to be a useful method for mRNA quantification, where high performance liquid chromatography (HPLC) with a UV detector was employed for detecting PCR products. 5-7)CYP2D6 with genetic polymorphism is involved in the oxidative metabolism of many drugs including neuroleptics, tricyclic antidepressants, selective serotonin reuptake inhibitors, b-adrenoreceptor blockers, and antiarrhythmics. 8,9) Although the genotyping of CYP2D6 is useful for assessing drug efficacy and adverse effects, it is hard to conduct routine clinical measurement because of the many single nucleotide polymorphisms (SNPs). Since blood is a readily accessible tissue, an appealing concept should be to use peripheral blood leukocytes (PBL) as a surrogate for CYP activities that manifest in internal organs. [10][11][12] In the present study, we developed a sensitive and quantitative RT-PCR-HPLC method for the measurement of CYP2D6 mRNA in PBL. ExperimentalPreparation of RNA Standards for RT-PCR wt 2D6, the standard cDNA containing 456 bp of CYP2D6 DNA (nt 1087-1542), was cloned to pBluescript KS (ϩ) vector by PCR from cDNA Library Human Liver (Takara Bio, Japan), and designated as pKSp2D6. For preparation of the internal standard, is 2D6, 68 bp (nt 1107-1153) was deleted from pKSp2D6 using PCR, and named as pKSd2D. The primers used for amplification of wt 2D6 were CGAGCAAGCTT-Ex7F, 5Ј-CGAGCAAGCTTGGAGATCG-ACGACGTGATAG-3Ј and AGGTCGTCGAC-Ex9R, 5Ј-AGGTCGTCGA-CACCAGGAAAGCAAAGACACC-3Ј. The primers used for amplification of is 2D6 were CGAGCAAGCTT-Ex7F D46, 5Ј-CGAGCAAGCTTGGA-GATCGACGACGTGATAGGTGCAGCGCTTTGGGGACGACAT-3Ј and AGGTCGTCGAC-Ex9R, 5Ј-AGGTCGTCGACACCAGGAAAGCAAA-GACACC-3Ј. The standard RNAs, wt 2D6 and is 2D6, were prepared using Riboprobe ® in vitro Transcription Systems (Promega, U.S.A.) from pKSp2D6 and pKSd2D6, respectively. The RNAs were treated with RNasefree DNase (QIAGEN, Netherlands) and purified by QIAamp ® RNA Blood Mini (QIAGEN). The RNA standards were diluted to the concentration of 1 pg/ml and stored in aliquots at Ϫ80°C until use.Subjects The study was explained to 8 healthy volunteers (2 male and 6 female) and written informed consent was obtained from each. The study was approved by the Institutional Review Board of the Faculty of Pharmaceutical Sciences, Teikyo University an...
Considerable progress has been made in the development of soft x-ray laser since 19851-2). Until now a powerful laser of single or double pulses has often been used to produce high density plasma. Recently we observed ASE (amplified spontaneous emission) in some lines of Al X and Al XI in a recombining plasma produced by a very low power laser of only 2 J which was composed of a train of many low power pulses separated by about 350 picosecond3). To understand more clearly the effect of multipulse irradiation on the production of laser plasma and x-ray emissions including ASE, in this paper we constructed a pulse train forming system and investigated the emitted soft x-ray spectra for various pulse trains. Our final goal is to develop a table top x-ray laser in the water window wavelength.
Amplified spontaneous emissions of Li-like ions were observed in a recombining Si plasma which was produced by a Nd-glass laser of table-top size. The small signal gain coefficient of Si XII 3d-5f (88.84 Å) was evaluated at 0.9 cm-1, and the emissions of Si XII 3d-4f (130.02 Å) and 3p-5d (87.36 Å) transitions show small amplifications. The driver laser pulse has a time envelope of 4 ns-FWHM (full width at half maximum) and contains several short pulses of about 120 ps-FWHM with time intervals of about 350 ps. The average irradiation power density of a short pulse was estimated to be only 5×1011 W/cm2.
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