Engraftment and retroviral marking of CD34+ and CD34+CD38- human hematopoietic progenitors assessed in immune-deficient mice

Retroviral-mediated transduction of human hematopoietic stem cells to provide a lifelong supply of corrected progeny remains the most daunting challenge to the success of human gene therapy. The paucity of assays to examine transduction of pluripotent human stem cells hampers progress toward this goal. By using the beige/nude/xid (bnx)/hu immune-deficient mouse xenograft system, we compared the transduction and engraftment of human CD34+ progenitors with that of a more primitive and quiescent subpopulation, the CD34+CD38- cells. Comparable extents of human engraftment and lineage development were obtained from 5 x 10(5) CD34+ cells and 2,000 CD34+CD38- cells. Retroviral marking of long-lived progenitors from the CD34+ populations was readily accomplished, but CD34+CD38- cells capable of reconstituting bnx mice were resistant to transduction. Extending the duration of transduction from 3 to 7 days resulted in low levels of transduction of CD34+CD38- cells. Flt3 ligand was required during the 7-day ex vivo culture to maintain the ability of the cells to sustain long-term engraftment and hematopoiesis in the mice.     

T lymphocytes with a normal ADA gene accumulate after transplantation of transduced autologous umbilical cord blood CD34+ cells in ADA-deficient SCID neonates

Adenosine deaminase-deficient severe combined immunodeficiency was the first disease investigated for gene therapy because of a postulated production or survival advantage for gene-corrected T lymphocytes, which may overcome inefficient gene transfer. Four years after three newborns with this disease were given infusions of transduced autologous umbilical cord blood CD34+ cells, the frequency of gene-containing T lymphocytes has risen to 1-10%, whereas the frequencies of other hematopoietic and lymphoid cells containing the gene remain at 0.01-0.1%. Cessation of polyethylene glycol-conjugated adenosine deaminase enzyme replacement in one subject led to a decline in immune function, despite the persistence of gene-containing T lymphocytes. Thus, despite the long-term engraftment of transduced stem cells and selective accumulation of gene-containing T lymphocytes, improved gene transfer and expression will be needed to attain a therapeutic effect.     

Aortic dissection in a patient receiving chemotherapy for Hodgkin's disease--a case report

Cancer chemotherapy is associated with a wide range of vascular toxicities, which may be related to endothelial cell damage by these agents. The authors describe a patient with Hodgkin's disease who developed an atypical aortic dissection while receiving MOPP/ABV chemotherapy (nitrogen mustard, vincristine, procarbazine, prednisone, doxorubicin, bleomycin, and vinblastine). They would place aortic dissection on the list of potential vascular complications associated with antineoplastic agents.     

The Ultralight Steel Auto Body program completes phase I

The UltraLight Steel Auto Body program was initiated by the American Iron and Steel Institute and the International Iron and Steel Institute. The purpose of the program is to demonstrate the advantages of steel for reducing the weight of the automotive body-in-white. The holistic design approach—considering the body structure, materials, manufacturing methods, and engineering analysis together—has yielded a design for a body-in-white that is about 25 percent lighter than a typical vehicle produced at present, with improved structural performance. The materials and manufacturing processes required for the design are either in use or “near-reach” technologies.     

Creep mechanisms in beryllium

Observations of beryllium samples which have been creep tested between 922 K and 1422 K indicate that creep behavior is controlled by the relative strengths of the grain boundaries and the matrix. Since creep deformation can occur predominantly by grain boundary sliding or entirely by deformation within the grains, the creep strength was found to be controlled by the weaker of the two features. Low melting phases containing aluminum and silicon which formed along the grain boundaries acted as stress concentrations which favored localized grain boundary deformation, and recrystallization. Creep resistance was found to drop markedly when the BeO content was reduced substantially below 1 pct.     

TV requirements for manipulation in space

An analytical and experimental study was conducted to specify a video system for remote manipulation in space. An operator function analysis identified two basic characteristics, work volume and element relationship, which define 4 manipulation tasks chosen for examination. A visual function analysis developed a set of elemental scene parameters which grouped the visual dimensions into major areas of influence. Simulation testing was conducted with a 4 degree-of-freedom motion frame which allowed an operator to perform the manipulation tasks. Four video systems were included in the simulation testing: a black and white and a color monoscopic system, a stereoscopic system, and a black and white 2-view system. A sequential experimental plan first provided an overall analysis of the effects of tasks, scene parameters and video systems. This was followed by a detailed experimental examination of the critical dimensions identified in the first experiment. Results are discussed in terms of a recommended TV system.     

Clean intermittent catheterization for home management in children with myelomeningocele

Sixty-two children with myelomeningocele and neurogenic bladders have been managed with clean intermittent catheterization (CIC) at home for periods of 6 to 42 months. Abnormal upper urinary tracts improved or stabilized in 83 per cent of the children. There was no renal deterioration in children who began with normal upper tracts while on CIC. Bacteruria was common, but clinically not a problem, unless ureterovesical reflux was present. Urinary control was improved in 66 per cent of the children. CIC is an effective and safe method to regularly empty the urinary bladder and is an alternative to supravesical urinary diversion in many children with neurogenic bladders.     

Li/Li2 supersonic nozzle beam

The characterization of a lithium supersonic nozzle beam has been made using spectroscopic techniques. It is found that at a stagnation pressure of 5.3 kPa (40 Torr) and a nozzle throat diameter of 0.4 mm the ground state vibrational population of Li(2) can be described by a Boltzmann distribution with T(v)= (195+/-30) K. The rotational temperature is found to be T(r)= (70+/-20) K by band shape analysis. Measurements by quadrupole mass spectrometer indicate that approximately 10-mol. % Li(2) dimers are present far downstream for an oven body temperature of 1370 K in the supersonic nozzle expansion. This measured mole fraction is in agreement with the existing dimerization theory.     

Parallel fabrication of RNA microarrays by mechanical transfer from a DNA master

A new method for fabrication of RNA microarrays is described. The approach involves cohybridization of a short, biotinylated DNA oligonucleotide and an RNA probe sequence to DNA templates spotted onto a master array. Next, the short DNA sequence and the RNA probe are linked using a T4 DNA ligase. Finally, a poly(dimethylsiloxane) (PDMS) monolith modified on the surface with streptavidin is brought into conformal contact with the master array. This results in binding of the biotinylated DNA/RNA oligonucleotides to the PDMS surface. When the two substrates are mechanically separated, the DNA/RNA oligonucleotides transfer to the PDMS replica, and the DNA oligonucleotides remaining on the master array are ready to template another RNA replica array. This sequence can be repeated for at least 18 cycles using a single master array. RNA arrays consisting of up to three different oligonucleotide sequences and consisting of up to 2500 individual approximately 70 microm spots have been prepared.