A point mutation at codon 13 of the N-ras oncogene in myelodysplastic syndrome

Patients with a myelodysplastic syndrome (MDS) which has a risk of leukaemic change exhibit a variable clinical course. It has been suggested that the development of leukaemia in patients with MDS may be related to chromosomal abnormalities or genetic alterations: somatic mutation of the N-ras gene is now considered to be a critical step in the genetic basis of human leukaemogenesis. Here we report that DNAs of bone-marrow cells from three out of eight patients with MDS contained an activated N-ras oncogene, as detected by an in vivo selection assay in nude mice with transfected NIH 3T3 cells. Molecular analysis revealed the same single nucleotide substitution at codon 13 in all three transforming N-ras genes. Each of the three patients showed a progression of the disease and a resulting leukaemic change within the following year. Our observation of the mutation at codon 13 in leukaemic cell DNAs from all three cases suggests that activation of the N-ras gene is important in the development of leukaemia in some MDS cases.     

Species-specific differences in the mitogenic activity of heparin-binding growth factors in the sera of various mammals.

Sera from different mammalian species displayed great differences in mitogenic activity, as measured by stimulation of DNA synthesis in BALB/c 3T3 cells (3T3 cells). Among the sera examined, fetal bovine serum was least active, and increasing activity was detected in calf serum, human serum, rat serum and mouse serum, in that order. Rat and mouse sera exhibited extremely high mitogenic activity with 3T3 cells, but when TIG-1 human fetal lung fibroblasts were used for the DNA assay instead, the activity levels of all of the sera were lower, and the differences between them were smaller. To determine the reasons for these differences, the heparin-binding growth factors in each serum were separated on a heparin affinity column. Five peaks of DNA-stimulating activity were obtained. Three of these were found in all sera examined, with both 3T3 cells and TIG-1 cells. Two other peaks were found only with 3T3 cells; one was peculiar to rat and mouse sera, with extremely high activity in the rat, and the other was specific to fetal serum. The dependence of the activity of these peaks on the cells used for the test was confirmed using normal rat lung fibroblasts and immortalized rat kidney cells. These findings adequately explain the species-specific differences in mitogenic activity of whole sera, and the variation in activity depending on the cells used for assay of DNA synthesis.     

Generation and characterization of a fairly stable triplet carbene

Most molecules are held together by covalent bonds-electron pairs jointly shared by the two atoms that are linked by the bond. Free radicals, in contrast, have at least one unpaired electron. In the case of carbon-based radicals, the carbon atom at the radical centre no longer makes four bonds with other atoms as it would do in its normal, tetravalent state. The presence of unpaired electrons renders such radicals highly reactive, so they normally occur only as transient intermediates during chemical reactions. But the discovery by Gomberg in 1900 of triphenylmethyl, the first relatively stable free radical containing a central trivalent carbon atom, illustrated that radicals with suitable geometrical and electronic structures can be stable. Compounds containing a divalent carbon atom that uses only two of its four valence electrons for bonding are usually less stable than Gomberg-type radicals with trivalent carbon. Although the role of these so-called carbenes in chemical reactions has long been postulated, they were unambiguously identified only in the 1950s. More recently, stable carbenes have been prepared, but the singlet state of these molecules, with the two nonbonding valence electrons paired, means that they are not radicals. Carbenes in the second possible electronic state, the triplet state, are radicals: the two nonbonding electrons have parallel spins and occupy different orbitals. Here we report the preparation and characterization of a triplet carbene, whose half-life of 19 minutes at room temperature shows it to be significantly more stable than previously observed triplet carbenes.     

A single-photon detector in the far-infrared range

The far-infrared region (wavelengths in the range 10 microm-1 mm) is one of the richest areas of spectroscopic research, encompassing the rotational spectra of molecules and vibrational spectra of solids, liquids and gases. But studies in this spectral region are hampered by the absence of sensitive detectors--despite recent efforts to improve superconducting bolometers, attainable sensitivities are currently far below the level of single-photon detection. This is in marked contrast to the visible and near-infrared regions (wavelengths shorter than about 1.5 microm), in which single-photon counting is possible using photomultiplier tubes. Here we report the detection of single far-infrared photons in the wavelength range 175-210 microm (6.0-7.1 meV), using a single-electron transistor consisting of a semiconductor quantum dot in high magnetic field. We detect, with a time resolution of a millisecond, an incident flux of 0.1 photons per second on an effective detector area of 0.1 mm2--a sensitivity that exceeds previously reported values by a factor of more than 10(4). The sensitivity is a consequence of the unconventional detection mechanism, in which one absorbed photon leads to a current of 10(6)-10(12) electrons through the quantum dot. By contrast, mechanisms of conventional detectors or photon assisted tunnelling in single-electron transistors produce only a few electrons per incident photon.     

Testing <Emphasis Type="Italic">k</Emphasis>-edge-connectivity of digraphs

This paper presents an algorithm that tests whether a given degree-bounded digraph is k-edge-connected or ɛ-far from k-edge-connectivity. This is the first testing algorithm for k-edgeconnectivity of digraphs whose running time is independent of the number of vertices and edges. A digraph of n vertices with degree bound d is ɛ-far from k-edge-connectivity if at least ɛdn edges have to be added or deleted to make the digraph k-edge-connected, preserving the degree bound. Given a constant error parameter ɛ and a degree bound d, our algorithm always accepts all k-edge-connected digraphs and rejects all digraphs that is ɛ-far from k-edge-connectivity with probability at least 2/3. It runs in $ O\left( {d\left( {\frac{c} {{\varepsilon d}}} \right)^k log\frac{1} {{\varepsilon d}}O} \right) $ O\left( {d\left( {\frac{c} {{\varepsilon d}}} \right)^k log\frac{1} {{\varepsilon d}}O} \right) (c > 1 is a constant) time when input digraphs are restricted to be (k-1)-edge connected and runs in $ O\left( {d\left( {\frac{{ck}} {{\varepsilon d}}} \right)^k log\frac{k} {{\varepsilon d}}O} \right) $ O\left( {d\left( {\frac{{ck}} {{\varepsilon d}}} \right)^k log\frac{k} {{\varepsilon d}}O} \right) (c > 1 is a constant) time for general digraphs.