INCIDENCE MATRIX APPROACH FOR CALCULATING READINESS LEVELS

Contemporary system maturity assessment approaches have failed to provide robust quantitative system evaluations resulting in increased program costs and developmental risks.Standard assessment metrics,such as Technology Readiness Levels（TRL）,do not sufficiently evaluate increasingly complex systems.The System Readiness Level（SRL）is a newly developed system development metric that is a mathematical function of TRL and Integration Readiness Level（IRL） values for the components and connections of a particular system.SRL acceptance has been hindered because of concerns over SRL mathematical operations that may lead to inaccurate system readiness assessments.These inaccurate system readiness assessments are called readiness reversals.A new SRL calculation method using incidence matrices is proposed to alleviate these mathematical concerns.The presence of SRL readiness reversal is modeled for four SRL calculation methods across several system configurations.Logistic regression analysis demonstrates that the proposed Incidence Matrix SRL（IMSRL）method has a decreased presence of readiness reversal than other approaches suggested in the literature.Viable SRL methods will foster greater SRL adoption by systems engineering professionals and will support system development risk reduction goals.     

Femtosecond X-ray protein nanocrystallography

X-ray crystallography provides the vast majority of macromolecular structures, but the success of the method relies on growing crystals of sufficient size. In conventional measurements, the necessary increase in X-ray dose to record data from crystals that are too small leads to extensive damage before a diffraction signal can be recorded. It is particularly challenging to obtain large, well-diffracting crystals of membrane proteins, for which fewer than 300 unique structures have been determined despite their importance in all living cells. Here we present a method for structure determination where single-crystal X-ray diffraction 'snapshots' are collected from a fully hydrated stream of nanocrystals using femtosecond pulses from a hard-X-ray free-electron laser, the Linac Coherent Light Source. We prove this concept with nanocrystals of photosystem I, one of the largest membrane protein complexes. More than 3,000,000 diffraction patterns were collected in this study, and a three-dimensional data set was assembled from individual photosystem I nanocrystals (～200?nm to 2?μm in size). We mitigate the problem of radiation damage in crystallography by using pulses briefer than the timescale of most damage processes. This offers a new approach to structure determination of macromolecules that do not yield crystals of sufficient size for studies using conventional radiation sources or are particularly sensitive to radiation damage.     

A large population of 'Lyman-break' galaxies in a protocluster at redshift z approximately 4.1

The most massive galaxies and the richest clusters are believed to have emerged from regions with the largest enhancements of mass density relative to the surrounding space. Distant radio galaxies may pinpoint the locations of the ancestors of rich clusters, because they are massive systems associated with 'overdensities' of galaxies that are bright in the Lyman-alpha line of hydrogen. A powerful technique for detecting high-redshift galaxies is to search for the characteristic 'Lyman break' feature in the galaxy colour, at wavelengths just shortwards of Lyalpha, which is due to absorption of radiation from the galaxy by the intervening intergalactic medium. Here we report multicolour imaging of the most distant candidate protocluster, TN J1338-1942 at a redshift z approximately 4.1. We find a large number of objects with the characteristic colours of galaxies at that redshift, and we show that this excess is concentrated around the targeted dominant radio galaxy. Our data therefore indicate that TN J1338-1942 is indeed the most distant cluster progenitor of a rich local cluster, and that galaxy clusters began forming when the Universe was only ten per cent of its present age.     

Large fluctuations in speed on Greenland's Jakobshavn Isbrae glacier

It is important to understand recent changes in the velocity of Greenland glaciers because the mass balance of the Greenland Ice Sheet is partly determined by the flow rates of these outlets. Jakobshavn Isbrae is Greenland's largest outlet glacier, draining about 6.5 per cent of the ice-sheet area, and it has been surveyed repeatedly since 1991 (ref. 2). Here we use remote sensing data to measure the velocity of Jakobshavn Isbrae between 1992 and 2003. We detect large variability of the velocity over time, including a slowing down from 6,700 m yr(-1) in 1985 to 5,700 m yr(-1) in 1992, and a subsequent speeding up to 9,400 m yr(-1) by 2000 and 12,600 m yr(-1) in 2003. These changes are consistent with earlier evidence for thickening of the glacier in the early 1990s and rapid thinning thereafter. Our observations indicate that fast-flowing glaciers can significantly alter ice discharge at sub-decadal timescales, with at least a potential to respond rapidly to a changing climate.     

Genetic changes associated with floral adaptation restrict future evolutionary potential

A commonly accepted evolutionary principle is that adaptive change constrains the potential directions of future evolutionary change. One manifestation of this is Dollo's law, which states that character elimination is irreversible. Although the common occurrence of irreversibility has been documented by phylogenetic analyses of phenotypic transitions, little is known about the underlying causes of this phenomenon. One explanation for evolutionary irreversibility relies on the fact that many characteristics result from interactions between multiple gene products. Such characteristics may often be eliminated by inactivation of just one gene in the network. If they serve no other functions, other genes of the network are then free to accumulate mutations or evolve new functions. Evolutionary change after character loss results in the accumulation of redundant loss-of-function mutations. Such pathway degeneration makes it very unlikely that the characteristic will re-evolve, because multiple simultaneous mutations would be required. Here we describe what appear to be the initial stages of such degeneration in the anthyocyanin pigment pathway associated with an adaptive change from blue to red flowers in the morning glory Ipomoea quamoclit.     

Haematopoietic stem cells do not transdifferentiate into cardiac myocytes in myocardial infarcts

The mammalian heart has a very limited regenerative capacity and, hence, heals by scar formation. Recent reports suggest that haematopoietic stem cells can transdifferentiate into unexpected phenotypes such as skeletal muscle, hepatocytes, epithelial cells, neurons, endothelial cells and cardiomyocytes, in response to tissue injury or placement in a new environment. Furthermore, transplanted human hearts contain myocytes derived from extra-cardiac progenitor cells, which may have originated from bone marrow. Although most studies suggest that transdifferentiation is extremely rare under physiological conditions, extensive regeneration of myocardial infarcts was reported recently after direct stem cell injection, prompting several clinical trials. Here, we used both cardiomyocyte-restricted and ubiquitously expressed reporter transgenes to track the fate of haematopoietic stem cells after 145 transplants into normal and injured adult mouse hearts. No transdifferentiation into cardiomyocytes was detectable when using these genetic techniques to follow cell fate, and stem-cell-engrafted hearts showed no overt increase in cardiomyocytes compared to sham-engrafted hearts. These results indicate that haematopoietic stem cells do not readily acquire a cardiac phenotype, and raise a cautionary note for clinical studies of infarct repair.     

Identification of human brain tumour initiating cells

The cancer stem cell (CSC) hypothesis suggests that neoplastic clones are maintained exclusively by a rare fraction of cells with stem cell properties. Although the existence of CSCs in human leukaemia is established, little evidence exists for CSCs in solid tumours, except for breast cancer. Recently, we prospectively isolated a CD133+ cell subpopulation from human brain tumours that exhibited stem cell properties in vitro. However, the true measures of CSCs are their capacity for self renewal and exact recapitulation of the original tumour. Here we report the development of a xenograft assay that identified human brain tumour initiating cells that initiate tumours in vivo. Only the CD133+ brain tumour fraction contains cells that are capable of tumour initiation in NOD-SCID (non-obese diabetic, severe combined immunodeficient) mouse brains. Injection of as few as 100 CD133+ cells produced a tumour that could be serially transplanted and was a phenocopy of the patient's original tumour, whereas injection of 10(5) CD133- cells engrafted but did not cause a tumour. Thus, the identification of brain tumour initiating cells provides insights into human brain tumour pathogenesis, giving strong support for the CSC hypothesis as the basis for many solid tumours, and establishes a previously unidentified cellular target for more effective cancer therapies.