Genomic alterations in cultured human embryonic stem cells

Cultured human embryonic stem cell (hESC) lines are an invaluable resource because they provide a uniform and stable genetic system for functional analyses and therapeutic applications. Nevertheless, these dividing cells, like other cells, probably undergo spontaneous mutation at a rate of 10(-9) per nucleotide. Because each mutant has only a few progeny, the overall biological properties of the cell culture are not altered unless a mutation provides a survival or growth advantage. Clonal evolution that leads to emergence of a dominant mutant genotype may potentially affect cellular phenotype as well. We assessed the genomic fidelity of paired early- and late-passage hESC lines in the course of tissue culture. Relative to early-passage lines, eight of nine late-passage hESC lines had one or more genomic alterations commonly observed in human cancers, including aberrations in copy number (45%), mitochondrial DNA sequence (22%) and gene promoter methylation (90%), although the latter was essentially restricted to 2 of 14 promoters examined. The observation that hESC lines maintained in vitro develop genetic and epigenetic alterations implies that periodic monitoring of these lines will be required before they are used in in vivo applications and that some late-passage hESC lines may be unusable for therapeutic purposes.     

Evidence of power-law flow in the Mojave desert mantle

Studies of the Earth's response to large earthquakes can be viewed as large rock deformation experiments in which sudden stress changes induce viscous flow in the lower crust and upper mantle that lead to observable postseismic surface deformation. Laboratory experiments suggest that viscous flow of deforming hot lithospheric rocks is characterized by a power law in which strain rate is proportional to stress raised to a power, n (refs 2, 3). Most geodynamic models of flow in the lower crust and upper mantle, however, resort to newtonian (linear) stress-strain rate relations. Here we show that a power-law model of viscous flow in the mantle with n = 3.5 successfully explains the spatial and temporal evolution of transient surface deformation following the 1992 Landers and 1999 Hector Mine earthquakes in southern California. A power-law rheology implies that viscosity varies spatially with stress causing localization of strain, and varies temporally as stress evolves, rendering newtonian models untenable. Our findings are consistent with laboratory-derived flow law parameters for hot and wet olivine--the most abundant mineral in the upper mantle--and support the contention that, at least beneath the Mojave desert, the upper mantle is weaker than the lower crust.     

Peptide sequences of T-cell receptor delta and gamma chains are identical to predicted X and gamma proteins

Although most mature peripheral T lymphocytes express a major histocompatibility complex restricted, CD3-associated, antigen receptor (TCR) which has been well characterized, some T cells carry a different CD3-associated heterodimer on their surface. One of the two disulphide-linked chains of this putative second receptor, which in mice has relative molecular mass (Mr) 35,000 (35K), has been identified as a product of the group of gamma genes. The other chain, termed delta (Mr 45K in mice), is not as well characterized. Although gamma/delta-bearing cells are a minor subset among peripheral T lymphocytes, they are the only CD3+ cells in the thymus early in ontogeny. Taking advantage of these kinetics, we have generated gamma/delta-bearing hybridomas, using a new TCR alpha chain-negative variant of the AKR thymoma BW5147 as tumour parent, fetal thymocytes as normal cell partners, and an anti-CD3 monoclonal antibody (mAb) as screening reagent. Gamma and delta chains from one of these hybrids have been purified and partially sequenced. The sequences obtained indicate that delta is indeed identical to the polypeptide encoded by the recently described gene X, as suggested by Chien et al.     

Self-destructive cooperation mediated by phenotypic noise

In many biological examples of cooperation, individuals that cooperate cannot benefit from the resulting public good. This is especially clear in cases of self-destructive cooperation, where individuals die when helping others. If self-destructive cooperation is genetically encoded, these genes can only be maintained if they are expressed by just a fraction of their carriers, whereas the other fraction benefits from the public good. One mechanism that can mediate this differentiation into two phenotypically different sub-populations is phenotypic noise. Here we show that noisy expression of self-destructive cooperation can evolve if individuals that have a higher probability for self-destruction have, on average, access to larger public goods. This situation, which we refer to as assortment, can arise if the environment is spatially structured. These results provide a new perspective on the significance of phenotypic noise in bacterial pathogenesis: it might promote the formation of cooperative sub-populations that die while preparing the ground for a successful infection. We show experimentally that this model captures essential features of Salmonella typhimurium pathogenesis. We conclude that noisily expressed self-destructive cooperative actions can evolve under conditions of assortment, that self-destructive cooperation is a plausible biological function of phenotypic noise, and that self-destructive cooperation mediated by phenotypic noise could be important in bacterial pathogenesis.     

Immunoglobulin-like nature of the alpha-chain of a human T-cell antigen/MHC receptor

Although the receptor with which T cells bind specific antigen can, like immunoglobulin, distinguish between antigens which differ only slightly in structure, it is unique in recognizing antigen only in conjunction with one of the self proteins of the major histocompatibility complex (MHC restriction). The receptor was identified and characterized in mouse and man by using monoclonal antibodies to receptor idiotypes, and consists of two disulphide-linked polypeptides, and acidic alpha-chain and a neutral to slightly basic beta-chain. Peptide maps have shown that, like immunoglobulin, both chains vary for receptors of different specificities. T-cell-derived cDNA clones have recently been identified in mouse and man encoding immunoglobulin-like molecules. These were identified as derived from beta-chain genes through a partial N-terminal protein sequence of the beta-chain isolated from a human T-cell tumour. We have now purified the alpha- and beta-chains of the receptor of the human T-cell leukaemia line HPB-MLT, and have determined the amino acid sequence of several tryptic peptides derived from each chain. Our results further confirm that the previously reported cDNA clones encode beta-chains. The sequence of the alpha-chain peptides identify this as another immunoglobulin-like polypeptide chain. Particularly striking was an alpha-chain peptide with high homology to the conserved portion of the immunoglobulin J segment and T-cell receptor beta-chains. Surprisingly, the alpha-chain peptides show little similarity to the sequence predicted by two overlapping putative murine alpha-chain cDNA clones.     

Delayed triggering of the 1999 Hector Mine earthquake by viscoelastic stress transfer

Stress changes in the crust due to an earthquake can hasten the failure of neighbouring faults and induce earthquake sequences in some cases. The 1999 Hector Mine earthquake in southern California (magnitude 7.1) occurred only 20 km from, and 7 years after, the 1992 Landers earthquake (magnitude 7.3). This suggests that the Hector Mine earthquake was triggered in some fashion by the earlier event. But uncertainties in the slip distribution and rock friction properties associated with the Landers earthquake have led to widely varying estimates of both the magnitude and sign of the resulting stress change that would be induced at the location of the Hector Mine hypocentre-with estimates varying from -1.4 bar (ref. 6) to +0.5 bar (ref. 7). More importantly, coseismic stress changes alone cannot satisfactorily explain the delay of 7 years between the two events. Here we present the results of a three-dimensional viscoelastic model that simulates stress transfer from the ductile lower crust and upper mantle to the brittle upper crust in the 7 years following the Landers earthquake. Using viscoelastic parameters that can reproduce the observed horizontal surface deformation following the Landers earthquake, our calculations suggest that lower-crustal or upper-mantle flow can lead to postseismic stress increases of up to 1-2 bar at the location of the Hector Mine hypocentre during this time period, contributing to the eventual occurrence of the 1999 Hector Mine earthquake. These results attest to the importance of considering viscoelastic processes in the assessment of seismic hazard.