Studies on pluripotent hematopoietic stem cells (HSCs) have been hindered by lack of a positive marker, comparable to the CD34 marker of hematopoietic progenitor cells (HPCs). In human postnatal hematopoietic tissues, 0.1 to 0.5% of CD34(+) cells expressed vascular endothelial growth factor receptor 2 (VEGFR2, also known as KDR). Pluripotent HSCs were restricted to the CD34+KDR+ cell fraction. Conversely, lineage-committed HPCs were in the CD34+KDR- subset. On the basis of limiting dilution analysis, the HSC frequency in the CD34+KDR+ fraction was 20 percent in bone marrow (BM) by mouse xenograft assay and 25 to 42 percent in BM, peripheral blood, and cord blood by 12-week long-term culture (LTC) assay. The latter values rose to 53 to 63 percent in LTC supplemented with VEGF and to greater than 95 percent for the cell subfraction resistant to growth factor starvation. Thus, KDR is a positive functional marker defining stem cells and distinguishing them from progenitors.
In several vertebrate species, Borna disease virus (BDV), the prototype of a new group of animal viruses, causes central nervous system disease accompanied by diverse behavioral abnormalities. Seroepidemiological data indicate that BDV may contribute to the pathophysiology of certain human mental disorders. This hypothesis is further supported by the detection of both BDV antigens and BDV RNA in peripheral blood mononuclear cells (PBMCs) of patients with psychiatric disorders and the isolation of BDV from such PBMCs.Here we describe serological and molecular epidemiological studies on psychiatric patients and healthy individuals from the area of Homburg, Germany. Using a novel Western blot (immunoblot) assay, we found a BDV seroprevalence of 9.6% among 416 neuropsychiatric patients, which is significantly higher than the 1.4% found among 203 healthy control individuals. Human sera displayed a prominent immunoreactivity against the virus nucleoprotein, the p40 antigen. Reverse transcriptase-mediated PCR analysis of RNA extracted from PBMCs of a subset of 26 of the neuropsychiatric patients revealed that 50% were BDV RNA positive. Three of the 13 BDV RNA-positive patients also had BDV-positive serology, whereas one patient with serum antibodies to BDV p40 antigen did not harbor detectable BDV RNA in PBMCs. BDV p40 and p24 sequences derived from human PBMCs exhibited both a high degree of inter-and intrapatient conservation and a close genetic relationship to animal-derived BDV sequences.
Cellular and molecular analysis of megakaryocytopoiesis has been hampered thus far by the lack of pure and abundant megakaryocyte (MK) cell populations. In this study, hematopoietic progenitor cells (HPCs), stringently purified from peripheral blood, were induced to megakaryocytic differentiation/maturation in serum-free liquid suspension culture treated with a growth factor cocktail (interleukin-3 [IL-3], c-kit ligand, and IL-6) and/or recombinant mpl ligand (mpIL). In particular, (1) the growth factor cocktail induced the growth of a 40% MK population, ie, 4 x 10(4) cells at day 0 generated 2 x 10(5) MK at terminal maturation; (2) further addition of mpIL increased the MK purity level to 80% with a final yield of 4 x 10(5) MKs; (3) treatment with mpIL alone resulted in a 97% to 99% MK population, with a mild increase of cell number (to 1.5 x 10(5) cells). In mpIL-supplemented culture, morphological evaluation indicated the presence of putative mononuclear MK precursors and then of mature polynucleated platelet- forming MKs, peaking at days 5 and 12, respectively. Membrane phenotype analysis showed a gradual decrease of CD34+ HPCs, coupled with an inverse increase of MK-specific antigens (eg, CD61/62/42b) starting before mature MK detection by morphology analysis. In situ hybridization showed the expression of MK-specific von Willebrand gene in both MK precursors and mature MKs. Furthermore, MKs synthesize and secrete low but significant amounts of both IL-6 and granulocyte- macrophage colony-stimulating factor. Comparative culture studies were performed on purified bone marrow CD34+/38hi or CD34+/38lo cells stimulated by mpIL alone. Both populations generated a highly enriched MK progeny (62% and 93% MKs at day 12 of culture, respectively) but showed either little or no proliferation. In conclusion, the purified peripheral blood HPC differentiation culture system allows for growth of a relatively large number of highly purified or “pure” megakaryocytic precursors and then mature MKs, thus providing an in vitro experimental tool to dissect the cellular and molecular basis of megakaryocytopoiesis.
Our data demonstrate that some MUC-1 epitopes are expressed on normal BM cells and particularly on cells of the erythroid lineage. Hence the application of anti-MUC-1 antibodies for disseminated tumor cell detection in BM or peripheral blood progenitor cells may provide false-positive results, and only carefully evaluated anti-MUC-1 antibodies (eg, HMFG-2) might be selected. Furthermore, MUC-1-targeted immunotherapy in cancer patients might be hampered by the suppression of erythropoiesis.
Using conductive and piezoforce microscopy, we reveal a complex picture of electronic transport at weakly conductive 109° domain walls in bismuth ferrite films. Even once initial ferroelectric stripe domains are changed/erased, persistent conductive paths signal the original domain wall position. The conduction at such domain wall “footprints” is activated by domain movement and decays rapidly with time, but can be re-activated by opposite polarity voltage. The observed phenomena represent true leakage conduction rather than merely displacement currents. We propose a scenario of hopping transport in combination with thermionic injection over interfacial barriers controlled by the ferroelectric polarization.
In-situ synchrotron x-ray diffraction was performed during the growth of BaTiO3 thin films on SrTiO3 substrates using both off-axis RF magnetron sputtering and pulsed laser deposition techniques. It was found that the films were ferroelectric during the growth process, and the presence or absence of a bottom SrRuO3 electrode played an important role in the growth of the films. Pulsed laser deposited films on SrRuO3 displayed an anomalously high tetragonality and unit volume, which may be connected to the previously predicted negative pressure phase of BaTiO3.
Using the model system of ferroelectric domain walls, we explore the effects of long-range dipolar interactions and periodic ordering on the behavior of pinned elastic interfaces. In piezoresponse force microscopy studies of the characteristic roughening of intrinsic 71 • stripe domains in BiFeO3 thin films, we find unexpectedly high values of the roughness exponent ζ = 0.74 ± 0.10, significantly different from those obtained for artificially written domain walls in this and other ferroelectric materials. The large value of the exponent suggests that a random field-dominated pinning, combined with stronger disorder and strain effects due to the step-bunching morphology of the samples, could be the dominant source of pinning in the system. [6]. Ferroelectric domain walls provide a useful model system in which many aspects of such glassy behavior can be readily accessed [7]. Previous studies of roughening, nonlinear dynamics, and aging have focused primarily on individual domain walls in uniaxial materials [8]. However, a particularly interesting experimental and theoretical challenge is posed by systems where coupled ferroic orders (such as ferroelectricity and ferroelasticity, or ferroelectricity and (anti)ferromagnetism [9]), as well as long-range interactions could lead to morecomplex behavior.Room-temperature multiferroic BiFeO 3 is an excellent candidate for investigating such phenomena. In this perovskite, polarization orientation along the eight pseudocubic [111] axes gives rise to three domain wall types (180 • , purely ferroelectric, and 71 • , 109 • , also ferroelastic), with magnetoelectric coupling between the ferroelectric and antiferromagnetic orders [10]. In addition, unusual domain wall functionalities [11,12] hold promise for future nanoelectronic applications [13,14]. BiFeO 3 films with specific polarization orientations, and domain structures ranging from nanoscale, sometimes fractallike "bubbles" to well-defined stripes can be obtained by adapting the deposition conditions and substrate [15][16][17]. Artificial domains can also be written by a biased scanning probe microscopy (SPM) tip, although this procedure can introduce significant electrochemical changes [18]. Intrinsic stripe domains follow standard LandauLifshitz-Kittel scaling of domain period w ∼ h 1/2 with the sample thickness h [19], while in samples with fractal bubble domains, a modified 0.59 exponent and apparent one-dimensional roughening of artificial domains were observed [20].Previous studies considered the domain walls as individual interfaces weakly pinned by disorder, with monoaffine roughness scaling characterized by a singlevalued roughness exponent ζ, dependent only on the disorder universality class and the system dimensionality. However, the heterogeneous disorder of ferroelectric thin films, with local universality class fluctuations and strong pinning [21], has recently been shown to lead to a breakdown of monoaffinity [22]. Moreover, in periodic systems interactions between neighboring interfaces can limit roughen...
A novel model system was used to investigate the spread of infectious particles and live cells through the application of lasers commonly used in clinical medicine. Supernatants from a cell line producing recombinant retroviruses carrying a marker gene (neoR) were exposed to Er:YAG-laser beams. Aerosols were collected from various sites and distances from the point of laser impact and were analyzed by reverse transcriptionpolymerase chain reaction (RT-PCR) for neoR. In addition, a susceptible indicator cell line was used to investigate the presence of infectious virions in collected aerosols. To test the possibility of dissemination of viable cells, a cell line was laser irradiated, and the generated aerosols were analyzed for the presence of viable cells. The viral marker gene neoR could be detected in 16% (distance: 5.0-6.3 cm) to 59% (0.5-1.6 cm) of wells adjacent to the point of laser impact. The presence of infectious viruses in laser vapors conferring G418 resistance could be detected in 3% (distance 5.0-6.3 cm) to 20% (distance: 0.5-1.6 cm) of wells containing susceptible cells, and subsequent PCR analysis of isolated resistant clones revealed the presence of neoR-RNA and -DNA. Viable cells were detected in 40% (distance 0.7-3.6 cm) to 3% (distance 10.7-11.8 cm) of wells adjacent to the point of laser impact. These results demonstrate that laser vapors can contain infectious viruses, viral genes, or viable cells and may promote the spread of infections or tumor cell dissemination.
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