运用一步化体外转录—翻译系统筛查基因突变的新技术

报道了一种筛查基因突变的新技术－ＰＣＲ结合一步化体外转转录－翻译系统。该技术包括：（１）ＰＣＲ扩增靶ＤＮＡ片段，并使ＤＮＡ产物的一端或两端带上Ｔ７Ｔ１序列；（Ｔ１：翻译起始信号，ＣＣＡ ＣＣＡ ＴＧ）；（２）以核的为模板，利用一步化体外转录－翻译系统合成相应肽链；（３）用ＳＤＳ－ＰＡＧＥ及高分辨ｐＨ梯度ＰＡＧＥ（聚丙烯酰胺凝胶电泳）分离分析多肽产物，通过异常电泳图谱而发现基因突变，本文以已知突变的     

Bibliometric analysis and private research funding

Summary The year 2002 brought a successive funding change-over from until now institutional to programme oriented funding (POF) in the Helmholtz Association of German Research Centres (HGF). This way the 15 German research centres now have to generate their means successively from programmes of the research fields of the HGF (see: www.helmholtz.de) by competing with each other. This nucleus of the reform of the Association is being implemented based upon the opinion of international experts. In this context the evaluation of publications of individual research centres, resp. research groups will be playing an ever increasing part. This lecture will inform about the reformed, partially formalized system and first experiences therewith at the time of the first evaluations.     

Database of mutations in the p53 and APC tumor suppressor genes designed to facilitate molecular epidemiological analyses

Germline and somatic mutations in the p53 and APC genes contribute to neoplasia. The patterns of these and other acquired mutations in cancers reflect environmental mutagens and endogenous factors that contribute to carcinogenesis. Herein, we describe a database of almost 2,300 mutations in the p53 and APC genes published until September 1, 1993. In addition to cataloging the mutations, multiple fields of information have been added to facilitate future molecular epidemiological analyses of human cancer. The accuracy of the database has been checked by the present authors and, by soliciting feedback from the original corresponding authors. The strengths and limitations of the primary literature are discussed.     

Possibilities to improve the short-term dosing constancy of volumetric feeders

For some applications, e.g. processes with stoichiometric reactions or the mixing of powders in a static mixer, a high short-term dosing constancy is necessary. For the dosing of calcium carbonate with a median diameter of 2 μm, it is shown that the pulsation of a volumetric feeder can be reduced tremendously by the use of different attachments for the dosing tube. The best results were obtained with a self-developed rotating star attachment. The quantitative assessment of the efficiency was performed both in the time domain with the standard deviation and in the frequency domain with the fast Fourier transformation.     

Colleagues and Criticism.

The author discusses the need for serious criticism of ideas and research in psychology, noting how papers circulated to colleagues for comment are often returned untouched. Several suggestions are provided as possible antidotes for this audience apathy. The writer must inform his colleagues whether a paper is being circulated simply as information, for evaluation, or for criticism. Professional meetings can be improved to encourage serious criticism about a paper read at a convention. The use of mimeographed newsletters for areas of research may be another solution. Ultimately, the responsibility for editing a paper remains with the author, and the author must state in clear language the implications of his results if he expects others to read his paper thoughtfully. Although too much criticism in a young science is unhealthy, too little criticism is even worse. Controversy brings forth a clarification of ideas and a resolution of conflicting data. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Signaling via CD28 costimulates lymphokine production, but does not reverse unresponsiveness to interleukin-2 in anti-CD3 triggered Th1 cells

Previously, it has been described that the ability of murine Th1 cells to proliferate in response to exogenous interleukin (IL)-2 is blocked when these cells are exposed to immobilized anti-CD3 antibodies. In the present study we examined whether simultaneous triggering of the T cell antigen CD28 can prevent the induction of unresponsiveness to IL-2 in Th1 cells. We report that costimulation of Th1 cells with anti-CD28 monoclonal antibodies (mAb) did not overcome unresponsiveness to IL-2 induced by various amounts of immobilized anti-CD3 antibodies. However, stimulation with anti-CD28 mAb strongly augmented IL-2 and interferon-gamma production in anti-CD3-exposed Th1 cells. Thus, despite the fact that anti-CD28 mAb is a potent costimulus for lymphokine production, signaling through CD28 does not seem to be sufficient to trigger proliferation in Th1 cells activated via the T cell receptor. These data suggest the existence of at least three signals to trigger Th1 cell activation. The first is mediated by ligation of the T cell receptor. One cosignal, delivered by the CD28 molecule, leads to IL-2 production. A third, still undefined, signal is required for proliferation in response to IL-2.     

The (8-thiotheophyllinate) (triphenylphosphine) gold (I), (tTAuP): a new gold complex as an anticancer agent

In continuous Friend leukemia cell exposure to tTAuP, the IC50 was 0.2 microM whereas in cells exposed 15 or 60 min to tTAuP followed by 72 h in drug-free medium the IC50 was 2.2 and 1.3 microM respectively. A combination of tTAuP and cisplatin (CDDP) is shown to be more active than either agent alone. Intracellular accumulation studies analysed by SXRF have shown that to achieve a cytotoxic effect, large concentrations of gold are necessary to accumulate in both the nuclear and cytoplasmic fraction. However, at cytostatic doses (1 microM), tTAuP has no effect on the cell cycle but does affect DNA synthesis. At a higher dose, greater than that necessary to induce cytotoxicity, it produces DNA damage, as observed by alkaline elution method. When cells were exposed to toxic doses of tTAuP (10 microM) in the presence of albumin, cytotoxicity was significantly reduced. Similar results were obtained when cells were co-treated with L-cysteine, dithiothreitol or reduced gluthatione. Reduced cytotoxic effect can be related to the interaction with free thiol groups. According to these data it is concluded that tTAuP is highly effective in vitro; whether it is active in vivo remains to be determined.     

A yeast antiviral protein,SKI8, shares a repeated amino acid sequence pattern with β-subunits of G proteins and several other proteins

SKI8 is a yeast antiviral gene, essential for controlling the propagation of M double-stranded RNA (dsRNA) and thus for preventing virus-induced cytopathology. Our DNA sequence of SKI8 shows that it encodes a 397 amino acid protein containing two copies of a 31 amino acid repeat pattern first identified in mammalian β-transducin and Cdc4p of yeast. There are also four copies of this repeat in yeast Mak11p, necessary for M dsRNA propagation, and three copies in the putative product of the Dictyostelium AAC3 gene. Analysis of 36 cases of the repeat unit shows they have a consensus predicted structure: N–helix–sheet–turn–sheet–turn–sheet–helix–C.