Haematopoietic stem cells adopt mature haematopoietic fates in ischaemic myocardium

Under conditions of tissue injury, myocardial replication and regeneration have been reported. A growing number of investigators have implicated adult bone marrow (BM) in this process, suggesting that marrow serves as a reservoir for cardiac precursor cells. It remains unclear which BM cell(s) can contribute to myocardium, and whether they do so by transdifferentiation or cell fusion. Here, we studied the ability of c-kit-enriched BM cells, Lin- c-kit+ BM cells and c-kit+ Thy1.1(lo) Lin- Sca-1+ long-term reconstituting haematopoietic stem cells to regenerate myocardium in an infarct model. Cells were isolated from transgenic mice expressing green fluorescent protein (GFP) and injected directly into ischaemic myocardium of wild-type mice. Abundant GFP+ cells were detected in the myocardium after 10 days, but by 30 days, few cells were detectable. These GFP+ cells did not express cardiac tissue-specific markers, but rather, most of them expressed the haematopoietic marker CD45 and myeloid marker Gr-1. We also studied the role of circulating cells in the repair of ischaemic myocardium using GFP+-GFP- parabiotic mice. Again, we found no evidence of myocardial regeneration from blood-borne partner-derived cells. Our data suggest that even in the microenvironment of the injured heart, c-kit-enriched BM cells, Lin- c-kit+ BM cells and c-kit+ Thy1.1(lo) Lin- Sca-1+ long-term reconstituting haematopoietic stem cells adopt only traditional haematopoietic fates.     

A Maturing of Systems Thinking? Evidence from Three Perspectives

This paper reviews trends in systems theory/thinking from the 1970s to the early 2000s. It proposes a maturation of the field based on certain conceptual and methodological advances that have sought to liberate systems thinking from earlier strictures. An edited dialogue among three prominent systems thinkers from different systems schools—Merrelyn Emery, Bob Flood, and Eric Wolstenholme—provides evidence. Similarities and differences are identified, complementarities among the schools are derived and analyzed, and trajectories for future research are indicated.     

An age-colour relationship for main-belt S-complex asteroids

Asteroid collisions in the main belt eject fragments that may eventually land on Earth as meteorites. It has therefore been a long-standing puzzle in planetary science that laboratory spectra of the most populous class of meteorite (ordinary chondrites, OC) do not match the remotely observed surface spectra of their presumed (S-complex) asteroidal parent bodies. One of the proposed solutions to this perplexing observation is that 'space weathering' modifies the exposed planetary surfaces over time through a variety of processes (such as solar and cosmic ray bombardment, micro-meteorite bombardment, and so on). Space weathering has been observed on lunar samples, in Earth-based laboratory experiments, and there is good evidence from spacecraft data that the process is active on asteroid surfaces. Here, we present a measurement of the rate of space weathering on S-complex main-belt asteroids using a relationship between the ages of asteroid families and their colours. Extrapolating this age-colour relationship to very young ages yields a good match to the colour of freshly cut OC meteorite samples, lending strong support to a genetic relationship between them and the S-complex asteroids.     

A complementary transposon tool kit for Drosophila melanogaster using P and piggyBac

With the availability of complete genome sequence for Drosophila melanogaster, one of the next strategic goals for fly researchers is a complete gene knockout collection. The P-element transposon, the workhorse of D. melanogaster molecular genetics, has a pronounced nonrandom insertion spectrum. It has been estimated that 87% saturation of the approximately 13,500-gene complement of D. melanogaster might require generating and analyzing up to 150,000 insertions. We describe specific improvements to the lepidopteran transposon piggyBac and the P element that enabled us to tag and disrupt genes in D. melanogaster more efficiently. We generated over 29,000 inserts resulting in 53% gene saturation and a more diverse collection of phenotypically stronger insertional alleles. We found that piggyBac has distinct global and local gene-tagging behavior from that of P elements. Notably, piggyBac excisions from the germ line are nearly always precise, piggyBac does not share chromosomal hotspots associated with P and piggyBac is more effective at gene disruption because it lacks the P bias for insertion in 5' regulatory sequences.     

Substrate twinning activates the signal recognition particle and its receptor

Signal sequences target proteins for secretion from cells or for integration into cell membranes. As nascent proteins emerge from the ribosome, signal sequences are recognized by the signal recognition particle (SRP), which subsequently associates with its receptor (SR). In this complex, the SRP and SR stimulate each other's GTPase activity, and GTP hydrolysis ensures unidirectional targeting of cargo through a translocation pore in the membrane. To define the mechanism of reciprocal activation, we determined the 1.9 A structure of the complex formed between these two GTPases. The two partners form a quasi-two-fold symmetrical heterodimer. Biochemical analysis supports the importance of the extensive interaction surface. Complex formation aligns the two GTP molecules in a symmetrical, composite active site, and the 3'OH groups are essential for association, reciprocal activation and catalysis. This unique circle of twinned interactions is severed twice on hydrolysis, leading to complex dissociation after cargo delivery.     

Vinculin activation by talin through helical bundle conversion

Vinculin is a conserved component and an essential regulator of both cell-cell (cadherin-mediated) and cell-matrix (integrin-talin-mediated focal adhesions) junctions, and it anchors these adhesion complexes to the actin cytoskeleton by binding to talin in integrin complexes or to alpha-actinin in cadherin junctions. In its resting state, vinculin is held in a closed conformation through interactions between its head (Vh) and tail (Vt) domains. The binding of vinculin to focal adhesions requires its association with talin. Here we report the crystal structures of human vinculin in its inactive and talin-activated states. Talin binding induces marked conformational changes in Vh, creating a novel helical bundle structure, and this alteration actively displaces Vt from Vh. These results, as well as the ability of alpha-actinin to also bind to Vh and displace Vt from pre-existing Vh-Vt complexes, support a model whereby Vh functions as a domain that undergoes marked structural changes that allow vinculin to direct cytoskeletal assembly in focal adhesions and adherens junctions. Notably, talin's effects on Vh structure establish helical bundle conversion as a signalling mechanism by which proteins direct cellular responses.     

Aging of connective tissues

Conclusion The brief summary of our present knowledge on the aging of intercellular matrix macromolecules shows the progress which has been made since the original important discovery of Verzár, but it also points out the very considerable gaps which still have to be filled by continued research efforts in this theoretically and practically important area. It is no secret to anyone that most of the disabling and killing diseases of advanced societies concern connective tissues: arteriosclerosis, diabetes, pulmonary obstructive lung diseases, osteoarticular diseases and cancer itself are all age-dependent, so-called aging diseases. The interaction between intercellular matrix and cancer cells plays an important role in the spreading of the tumors. For these other major diseases, the direct involvement of intercellular matrix is well documented. It is therefore hoped that a better grasp of the basic mechanisms involved in these diseases will help us to understand the difference between pathology and aging per se.