Imaging of Rab5 activity identifies essential regulators for phagosome maturation

Efficient phagocytosis of apoptotic cells is crucial for tissue homeostasis and the immune response. Rab5 is known as a key regulator of the early endocytic pathway and we have recently shown that Rab5 is also implicated in apoptotic cell engulfment; however, the precise spatio-temporal dynamics of Rab5 activity remain unknown. Here, using a newly developed fluorescence resonance energy transfer biosensor, we describe a change in Rab5 activity during the engulfment of apoptotic thymocytes. Rab5 activity on phagosome membranes began to increase on disassembly of the actin coat encapsulating phagosomes. Rab5 activation was either continuous or repetitive for up to 10 min, but it ended before the collapse of engulfed apoptotic cells. Expression of a dominant-negative mutant of Rab5 delayed this collapse of apoptotic thymocytes, showing a role for Rab5 in phagosome maturation. Disruption of microtubules with nocodazole inhibited Rab5 activation on the phagosome membrane without perturbing the engulfment of apoptotic cells. Furthermore, we found that Gapex-5 is the guanine nucleotide exchange factor essential for Rab5 activation during the engulfment of apoptotic cells. Gapex-5 was bound to a microtubule-tip-associating protein, EB1, whose depletion inhibited Rab5 activation during phagocytosis. We therefore propose a mechanistic model in which the recruitment of Gapex-5 to phagosomes through the microtubule network induces the transient Rab5 activation.     

Identification of a ferrireductase required for efficient transferrin-dependent iron uptake in erythroid cells

The reduction of iron is an essential step in the transferrin (Tf) cycle, which is the dominant pathway for iron uptake by red blood cell precursors. A deficiency in iron acquisition by red blood cells leads to hypochromic, microcytic anemia. Using a positional cloning strategy, we identified a gene, six-transmembrane epithelial antigen of the prostate 3 (Steap3), responsible for the iron deficiency anemia in the mouse mutant nm1054. Steap3 is expressed highly in hematopoietic tissues, colocalizes with the Tf cycle endosome and facilitates Tf-bound iron uptake. Steap3 shares homology with F(420)H(2):NADP(+) oxidoreductases found in archaea and bacteria, as well as with the yeast FRE family of metalloreductases. Overexpression of Steap3 stimulates the reduction of iron, and mice lacking Steap3 are deficient in erythroid ferrireductase activity. Taken together, these findings indicate that Steap3 is an endosomal ferrireductase required for efficient Tf-dependent iron uptake in erythroid cells.     

Are 16‐month‐ahead forecasts useful? A directional analysis of Japanese GDP forecasts

Past literature casts doubt on the ability of long‐term macroeconomic forecasts to predict the direction of change. We re‐examine this issue using the Japanese GDP forecast data of 37 institutions, and find that their 16‐month‐ahead forecasts contain valuable information on whether the growth rate accelerates or not. Copyright © 2006 John Wiley _ Sons, Ltd.     

与环境保全并存的生业的可能性:水满村的事例

生活在海南岛水满村的黎族的传统生业主要由水田农耕、火田农耕陆稻、根茎类作物以及其他作物)、野生植物采集和野生动物狩猎构成。在村周围覆盖着大面积的草地,这种人工环境是定期焚烧的结果。然而自从 20 世纪 80 年代以来,由于政府环保政策的实施,这样的生业方式已经发生了巨变。焚烧被禁止,水田农耕也得到了强化。与此同时,人们也强化了水田可使用野生植物的利用。这样生业变化导致了村落周围二次植被的恢复,也改善了村民们日常饮食的营养摄入。从环境学或营养学上说,当今的水满村的农业生产也许是合理的生业。尽管不断变化的市场经济价值观可能会影响到这种生业的可持续性,但是这种生业还是可能成为未来亚洲各国农业的模范。     

一种在RTL测试模式生成中验证断言再用的方法

在对设计的功能验证中,断言常被用于检测设计错误.针对制造业的测试模式生成,提出了在寄存器传输层(RTL)用于无扫描设计的断言再用方法.这种方法减少了顺序自动测试码生成程序(ATPG)的搜索空间,因而能加快测试生成过程,增加故障覆盖率.通过实例分析,证明了该方法的可行性和效果.     

Bone marrow cells adopt the phenotype of other cells by spontaneous cell fusion

Recent studies have demonstrated that transplanted bone marrow cells can turn into unexpected lineages including myocytes, hepatocytes, neurons and many others. A potential problem, however, is that reports discussing such 'transdifferentiation' in vivo tend to conclude donor origin of transdifferentiated cells on the basis of the existence of donor-specific genes such as Y-chromosome markers. Here we demonstrate that mouse bone marrow cells can fuse spontaneously with embryonic stem cells in culture in vitro that contains interleukin-3. Moreover, spontaneously fused bone marrow cells can subsequently adopt the phenotype of the recipient cells, which, without detailed genetic analysis, might be interpreted as 'dedifferentiation' or transdifferentiation.     

Transferrin receptor is necessary for development of erythrocytes and the nervous system

Plasma iron circulates bound to transferrin (Trf), which solubilizes the ferric ion and attenuates its reactivity. Diferric Trf interacts with cell-surface Trf receptor (Trfr) to undergo receptor-mediated endocytosis into specialized endosomes. Endosomal acidification leads to iron release, and iron is transported out of the endosome through the activity of divalent metal transporter 1 (DMT1, formerly Nramp2), a transmembrane iron transporter that functions only at low pH. Trf and Trfr then return to the cell surface for reuse, completing a highly efficient cycle. Although the Trf cycle is assumed to be the general mechanism for cellular iron uptake, this has not been validated experimentally. Mice with hypotransferrinaemia (hpx) have little or no plasma Trf. They have severe anaemia, indicating that the Trf cycle is essential for iron uptake by erythroid cells. Other hpx tissues, however, are generally normal, and there is a paradoxical increase in intestinal iron absorption and iron storage. To test the hypothesis that the Trf cycle has unique importance for erythropoiesis, we disrupted the Trfr gene in mice. This results in elimination of the Trf cycle, but leaves other Trf functions intact. Mice lacking Trfr have a more severe phenotype than hpx mice, affecting both erythropoiesis and neurologic development. Furthermore, haploinsufficiency for Trfr results in impaired erythroid development and abnormal iron homeostasis.     

Autoradiographic studies on the rat embryos in the pre-implantation stages

Zusammenfassung Die Rattenblastocyste synthetisiert, kurz bevor sich die Membrana pellucida auflöst, Nukleinsäuren, wie aus der Aufnahme von Uridin durch die Zellkerne elektronenmikroskopisch und autoradiographisch gezeigt wurde. Bei verzögerter Implantation wird Nukleinsäure in geringerem Ausmass aufgebaut.     

Light-induced conversion of an insulating refractory oxide into a persistent electronic conductor

Materials that are good electrical conductors are not in general optically transparent, yet a combination of high conductivity and transparency is desirable for many emerging opto-electronic applications. To this end, various transparent oxides composed of transition or post-transition metals (such as indium tin oxide) are rendered electrically conducting by ion doping. But such an approach does not work for the abundant transparent oxides of the main-group metals. Here we demonstrate a process by which the transparent insulating oxide 12CaO x 7Al(2)O(3) (refs 7-13) can be converted into an electrical conductor. H(-) ions are incorporated into the subnanometre-sized cages of the oxide by a thermal treatment in a hydrogen atmosphere; subsequent irradiation of the material with ultraviolet light results in a conductive state that persists after irradiation ceases. The photo-activated material exhibits moderate electrical conductivity (approximately 0.3 S cm(-1)) at room temperature, with visible light absorption losses of only one per cent for 200-nm-thick films. We suggest that this concept can be applied to other main-group metal oxides, for the direct optical writing of conducting wires in insulating transparent media and the formation of a high-density optical memory.