Milieu-adopted in vitro and in vivo differentiation of mesenchymal tissues derived from different adult human CD34-negative progenitor cell clones

Adult mesenchymal stem cells with multilineage differentiation potentially exist in the bone marrow, but have also been isolated from the peripheral blood. The differentiation of stem cells after leaving their niches depends predominately on the local milieu and its new microenvironment, and is facilitated by soluble factors but also by the close cell-cell interaction in a three-dimensional tissue or organ system. We have isolated CD34-negative, mesenchymal stem cell lines from human bone marrow and peripheral blood and generated monoclonal cell populations after immortalization with the SV40 large T-antigen. The cultivation of those adult stem cell clones in an especially designed in vitro environment, including self-constructed glass capillaries with defined growth conditions, leads to the spontaneous establishment of pleomorphic three-dimensional cell aggregates ( spheroids) from the monoclonal cell population, which consist of cells with an osteoblast phenotype and areas of mineralization along with well-vascularized tissue areas. Modifications of the culture conditions favored areas of bone-like calcifications. After the transplantation of the at least partly mineralized human spheroids into different murine soft tissue sites but also a dorsal skinfold chamber, no further bone formation could be observed, but angiogenesis and neovessel formation prevailed instead, enabling the transplanted cells and cell aggregates to survive. This study provides evidence that even monoclonal adult human CD34-negative stem cells from the bone marrow as well as peripheral blood can potentially differentiate into different mesenchymal tissues depending on the local milieu and responding to the needs within the microenvironment. Copyright (C) 2005 S. Karger AG, Basel

Clinical and functional characterisation of a novel TNFRSF1A c.605T > A/V173D cleavage site mutation associated with tumour necrosis factor receptor-associated periodic fever syndrome (TRAPS), cardiovascular complications and excellent response to etanercept treatment.

Objectives: To study the clinical outcome, treatment response, T-cell subsets and functional consequences of a novel tumour necrosis factor (TNF) receptor type 1 (TNFRSF1A) mutation affecting the receptor cleavage site. Methods: Patients with symptoms suggestive of tumour necrosis factor receptor-associated periodic syndrome (TRAPS) and 22 healthy controls (HC) were screened for mutations in the TNFRSF1A gene. Soluble TNFRSF1A and inflammatory cytokines were measured by ELISAs. TNFRSF1A shedding was examined by stimulation of peripheral blood mononuclear cells (PBMCs) with phorbol 12-myristate 13-acetate followed by flow cytometric analysis (FACS). Apoptosis of PBMCs was studied by stimulation with TNFa in the presence of cycloheximide and annexin V staining. T cell phenotypes were monitored by FACS. Results: TNFRSF1A sequencing disclosed a novel V173D/ p.Val202Asp substitution encoded by exon 6 in one family, the c.194–14G.A splice variant in another and the R92Q/p.Arg121Gln substitution in two families. Cardiovascular complications (lethal heart attack and peripheral arterial thrombosis) developed in two V173D patients. Subsequent etanercept treatment of the V173D carriers was highly effective over an 18-month follow-up period. Serum TNFRSF1A levels did not differ between TRAPS patients and HC, while TNFRSF1A cleavage from monocytes was significantly reduced in V173D and R92Q patients. TNFa-induced apoptosis of PBMCs and T-cell senescence were comparable between V173D patients and HC. Conclusions: The TNFRSF1A V173D cleavage site mutation may be associated with an increased risk for cardiovascular complications and shows a strong response to etanercept. T-cell senescence does not seem to have a pathogenetic role in affected patients

The solar oxygen-isotopic composition: Predictions and implications for solar nebula processes

The outer layers of the Sun are thought to preserve the average isotopic and chemical composition of the solar system. The solar O-isotopic composition is essentially unmeasured, though models based on variations in meteoritic materials yield several predictions. These predictions are reviewed and possible variations on these predictions are explored. In particular, the two-component mixing model of Clayton and Mayeda (1984) (slightly revised here) predicts solar compositions to lie along an extension of the calcium-aluminum-rich inclusion (CAI) ^(16)O line between (δ^(18)O, δ^(17)O) = (16.4, 11.4)%0 and (12.3, 7.5)%0. Consideration of data from ordinary chondrites suggests that the range of predicted solar composition should extend to slightly lower δ^(18)O values. The predicted solar composition is critically sensitive to the solid/gas ratio in the meteorite-forming region, which is often considered to be significantly enriched over solar composition. A factor of two solid/gas enrichment raises the predicted solar (δ^(18)O, δ^(17)O) values along an extension of the CAI ^(16)O line to (33, 28)%0. The model is also sensitive to the nebular O gas phase. If conversion of most of the gaseous O from CO to H_2O occurred at relatively low temperatures and was incomplete at the time of CM aqueous alteration, the predicted nebular gas composition (and hence the solar composition) would be isotopically heavier along a slope 1/2 line. The likelihood of having a single solid nebular O component is discussed. A distribution of initial solid compositions along the CAI ^(16)O line (rather than simply as an end-member) would not significantly change the predictions above in at least one scenario. Even considering these variations within the mixing model, the predicted range of solar compositions is distinct from that expected if the meteoritic variations are due to non-mass-dependent fractionation. Thus, a measurement of the solar O composition to a precision of several permil would clearly distinguish between these theories and should clarify a number of other important issues regarding solar system formation

An operative gamma camera for sentinel lymph node procedure in case of breast cancer

Large field of view gamma cameras are widely used to perform lymphoscintigraphy in the sentinel lymph nodes (SLN) procedure in case of breast cancer. However, they are not specified for this application and their sizes do not enable their use in the operative room to control the excision of the all SLN. We present the results obtained with a prototype of a new mini gamma camera developed especially for the operative lymphoscintigraphy of the axillary area in case of breast cancer. This prototype is composed of 10 mm thick parallel lead collimator, a 2 mm thick GSO:Ce inorganic scintillating crystal from Hitachi and a Hamamatsu H8500 flat panel multianode (64 channels) photomultiplier tube (MAPMT) equipped with a dedicated electronics. Its actual field of view is 50 × 50mm2. The gamma interaction position in the GSO scintillating plate is obtained by calculating the center of gravity of the fired MAPMT channels. The measurements performed with this prototype demonstrate the usefulness of this mini gamma camera for the pre, per and post-operative identification of SLN's and how its complementary role with an intraoperative handheld gamma probe enables to improve the efficiency of this practice. A 100 × 100mm2 field of view camera designated to cover the entire axillary area is under investigation