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PubMed1996

Is blood a better source of allogeneic hematopoietic stem cells for use after radiation accidents?

Peripheral blood and cord blood are increasingly used as sources of hematopoietic stem cells. They offer several advantages over bone marrow-derived stem cell grafts and are expected to replace bone marrow transplantation for treatment of malignant hematological disorders and solid tumors. Since the benefit of bone marrow transplantation for treatment of victims of radiation accidents has been seriously questioned, it is reasonable to examine if these alternative stem cell sources may change future strategies. Cord blood is attractive since it is an immediately available and fully characterized allogeneic stem cell source. However, experience with transplants in children suggests that recovery times are too long to benefit a victim of a radiation accident. Because of their potential to rapidly restore hematopoiesis, peripheral blood stem cells may currently be the best stem cell source for a patient who is expected to die from the consequences of prolonged pancytopenia (and not from damage of nonhematopoietic tissue). However, before getting accepted as the optimal stem cell source, the risk of acute and chronic GVHD must be confirmed to be low and unrelated donors must be shown to accept the potential risk of treatment with mobilizing agents. Future strategies should aim at developing methods of transiently rather than permanently engrafting allogeneic stem cells, since essentially all patients exposed to high doses of irradiation are expected to recover autologous marrow function.

Blood Cells↗
PubMed1995

Peripheral blood CD34+ cells: method of purification and ex vivo expansion.

Peripheral blood stem cells (PBSC) are used increasingly for autotransplantation in the treatment of acute leukemia, lymphoma, multiple myeloma, solid tumors such as ovarian and breast carcinoma. They are collected by leukaphereses during rapid hematopoietic recovery, following cytotoxic chemotherapy with or without administration of hematopoietic growth factors. We studied the clonogenic and cytokine-mediated expansion potential of CD34+ cells from mobilized PBSC. Low density mononuclear cells were processed using the CEPRATE LC CD34 KIT (CellPro). CD34+ purified cells, were cultured in suspension with 6 combined hematopoietic growth factors (IL1beta, IL3, IL6 at 100 U/ml and G-CSF, GM-CSF and stem cell factor at 10 ng/ml of each) for up to four weeks. Every week, cells were counted and CFU-GM assay was performed in a methylcellulose based medium. We have analysed the percentage of cells bearing CD34, CD33, CD38, HLA-DR, CD45RA, CD45RO antigens. Our results showed, that CD34+ cells were obtained with a purity of 92 +/- 2.3% and a yield of 71 +/- 10.7%. The majority co-expressed CD33 (57.76 +/- 34.16%) and CD38 (62.2 +/- 34%) antigens. These culture conditions, are necessary to obtain a fold increase of nucleated cells (377 fold at week 4), of CFU-GM progenitors (41.2 fold at week 3) and of CD34+ cell absolute number (10 fold at week 1) with an important differentiation of progenitors in particular myeloid progenitors.

Antigens, CD34↗
PubMed1997

Separation and quantification of apolipoprotein B-48 and other apolipoproteins by dynamic sieving capillary electrophoresis.

Apolipoprotein-B-48 is a structural protein exclusively associated with post-prandial lipoproteins (chylomicrons). Apolipoprotein B-48 would be a useful marker to monitor the kinetics of chylomicrons in vivo, however, its quantitation is limited because of a low concentration in plasma and lack of specific antibodies. Dynamic sieving capillary electrophoresis (DSCE) has recently become widely available for the separation of nanomolar quantities of proteins by size and electrophoretic mobility. Here we describe the potential of DSCE to accurately quantitate apolipoprotein mass in one ml of plasma. Separation of human serum apolipoproteins was achieved through an uncoated fused silica glass capillary column with quantitation based on area response at 220 nm. The retention times of human apolipoprotein B-48, apolipoprotein B-100 and albumin were 8.96 min +/- 0.57%, 10.21 min +/- 0.72%, and 6.56 min +/- 0.4%, respectively (phase-standardized to internal reference). A significant correlation (r = 0.99) was observed between apolipoprotein concentrations and peak area response for mass ranges of 30-40 micrograms/ml. DSCE provides an alternative method for quantifying apolipoprotein B-48 and therefore, may be useful for studying postprandial lipoprotein metabolism.

Animals↗
PubMedDate not supplied

[Mobilization using incompatibility group P1 plasmids in strains of Pseudomonas pseudomallei and Pseudomonas mallei as potential vectors for DNA cloning].

The cells of Pseudomonas pseudomallei and Pseudomonas mallei have been shown to serve as recipients for the plasmid RSF1010 and its recombinant derivatives pVA1 and pVA4. The conjugative plasmids RP1 and pTH10 of the incompatibility group P1 are able to mobilize the nontransmissive vector plasmids for conjugation transfer into Pseudomonas pseudomallei and Pseudomonas mallei strains. The SmR determinant of the plasmid RSF1010 is expressed in the latter strains. These data makes the mentioned vector plasmids the candidates for DNA cloning in these strains.

Burkholderia pseudomallei↗
PubMed1994

Effect of low dose radiation on intracellular calcium and protein kinase C in lymphocytes.

It is first reported in the present paper that whole-body irradiation (WBI) with low dose X-rays could increase intracellular calcium ions ([Ca2+]i) and stimulate protein kinase C (PKC) activity of mouse lymphocytes. Following WBI of male Kunming mice with 75 mGy X-rays at a dose rate of 12.5 mGy/min the mobilization of [Ca2+]i with Con A in CD4+ and CD8+ Cells in the thymus and spleen was potentiated and the amplitude of [Ca2+]i mobilization in thymocytes in response to anti-CD3 monoclonal antibody increased with time from 4 to 24 h following low dose radiation. The PKC activity in the homogenate of spleen was markedly stimulated 12 h after WBI with 75 mGy, reaching its peak value at 24-48 h and coming down to lower than normal on day 7. However, the PKC activity in the separated T lymphocytes reached its peak value at 12 h and that in the B lymphocytes reached its peak value on day 4, both coming down to below control on day 7. The implications of this facilitation of signal transduction in T lymphocytes in the mechanism of immunoenhancement after low dose radiation were discussed.

Animals↗
PubMed1989

Binding of neomycin to phosphatidylinositol 4,5-bisphosphate (PIP2).

Schacht (Schacht, J. (1976) J. Neurochem. 27, 1119-1124) demonstrated that neomycin, an aminoglycoside antibiotic, binds with high affinity to phosphatidylinositol 4,5-bisphosphate (PIP2). We investigated the binding of neomycin to PIP2 by making electrophoretic mobility measurements with multilamellar bilayer vesicles and surface potential measurements with monolayers. The bilayers and monolayers were formed from mixtures of PIP2 and egg phosphatidylcholine (PC) in 0.1 M KCl at pH 7. Neomycin does not bind to PC; 10(-3) M neomycin affects neither the zeta potential of PC vesicles nor the surface potential of PC monolayers. In contrast, 10(-6) M neomycin reduces the magnitude of the zeta potential of PC/PIP2 vesicles (5, 9, and 17 mol% PIP2) and the surface potential of monolayers (17 mol% PIP2) to less than 50% of their initial values. The electrophoretic mobility results indicate that neomycin forms an electroneutral complex with PIP2; high concentrations (greater than 10(-4) M) of neomycin reduce the zeta potential of the PC/PIP2 vesicles to zero. We could describe our data with the Gouy-Chapman-Stern theory assuming the intrinsic association constant of the 1:1 neomycin-PIP2 complex is 10(5) M-1. Neomycin is widely used in cell biology to interfere with the generation of second messengers; we discuss the relevance of our results to these studies. Specifically, 10(-6) M neomycin binds greater than 50% of the PIP2 in a bilayer or monolayer but 10(-5)-10(-3) M neomycin is required to affect the turnover of PIP2 in permeabilized platelets, mast cells, and sea urchin eggs. This result is consistent with a hypothesis that most of the PIP2 in the inner leaflet of these plasma membranes is not accessible to neomycin because it is associated with proteins.

Electrophoresis↗
PubMed1978

The adsorption of divalent cations to phosphatidylcholine bilayer membranes.

Electrophoretic mobility and 31P NMR measurements were combined to test whether the combination of the Henry, Boltzmann and Grahame equations is capable of describing the adsorption of divalent cations of phosphatidylcholine membranes. Cobalt was chosen for this study because, of all the common divalent cations, its effects on the 31P NMR spectrum of phosphatidylcholine membranes are easiest to interpret. Both the 31P NMR data on the adsorption of cobalt and the zeta potential data calculated from the electrophoretic mobility in the presence of cobalt are well described by the combination of these three equations. Electrophoretic mobility measurements were also performed with a number of other divalent cations and the zeta potentials were, in all cases, well described by the combination of these three equations. The binding deduced from such measurements decreases in the sequence: Mn2+, Mg2+, Ca2+, Co2+, Ni2+, Sr2+, Ba2+. If we assume that a lipid molecule occupies an area of 60 A2 and that there is a 1 : 1 stoichiometry for the binding of the divalent ions to phosphatidylcholine, the dissociation constants are, respectively: 0.3, 1.0, 1.0, 1.2, 1.2, 2.8, 3.6 M.

Adsorption↗
PubMed2006

Pharyngolaryngoesophagectomy using the thoracoscopic approach.

BACKGROUND: Thoracoscopic mobilization of the esophagus for pharyngolaryngoesophagectomy allows dissection under direct vision, and therefore it potentially results in fewer complications than conventional transhiatal mobilization. In this article we report our experience with this approach. It was also hypothesized that a learning curve existed and that results have improved over time. PATIENTS AND METHODS: From July 1994 until January 2004, 57 patients underwent pharyngolaryngoesophagectomy in our institution. Intraoperative events and postoperative outcome were prospectively documented, and long-term follow-up data were also studied. Results were compared between the first 30 patients and the last 27 patients. RESULTS: There were no significant differences between the two groups with respect to the various clinicopathological characteristics. There was no difference in the median thoracoscopic time between the first 30 and last 27 patients at 90 and 75 min, respectively, p = 0.18. For the complete procedure there was significantly less blood loss in the later group; median (range) blood loss 700 (164-3000) ml versus 400 (100-1200) ml, p = 0.002. Overall pulmonary complications occurred in 12 patients (40%) in the first group versus 13 (48%) in the second group, p = 0.6. The incidence of atrial arrhythmia was also similar, affecting 6 (20%) patients and 3 (11%), respectively, p = 0.47. Hospital mortality rates were 13.3% and 7.4%, p = 0.67. Two-year survival rates were no different (46% versus 45% p = 0.85). CONCLUSIONS: Although, subjectively, operating skills have improved over time, better results in the second half of this series could not be demonstrated clearly, likely because the operating surgeons had prior extensive experience in esophageal and thoracoscopic procedures.

Adult↗
PubMed2005

Endovascular stent graft for symptomatic mobile thrombus of the thoracic aorta.

Mobile thoracic aortic thrombus is a potential source of arterial embolism. Therapeutic management remains controversial. Systemic anticoagulation and various open surgical procedures are the commonly used therapeutic modalities. We report the successful primary treatment by endovascular stent graft of a mobile thoracic aortic thrombus that had caused visceral and peripheral embolism. Our case shows that endovascular stent-graft treatment is an effective, minimally invasive treatment of symptomatic mobile thoracic aortic thrombus.

Aged↗
PubMed2002

Land application technique for the treatment and disposal of sewage sludge.

Some of the major concerns when applying sewage sludge to land include the potential effect on pH and cation exchange capacity; the mobility and the accumulation of heavy metals in sludge treated soil; the potential of applying too much nutrients and the problems associated with odors and insects. The main objective of this study is to identify the effects of sewage sludge application on the physical and chemical properties of sludge treated soil. Sewage sludge was applied to soil at various rates ranging from 0 L/m2 to 341 L/m2. In order to simulate the natural environment, the study was carried out at a pilot treatment site (5.2 m x 6.7 m) in an open area, covered with transparent roofing material to allow natural sunlight to pass through. Simulated rain was applied by means of a sprinkler system. Data obtained from sludge treated soil showed that the pH values decreased when the application rates were increased and the application period prolonged. The effect of sewage sludge on cation exchange capacity was not so clear; the values obtained for every application rate of sewage sludge did not indicate any consistent behaviour. The mobility of heavy metals in soils treated with sludge were described by observing the changes in the concentration of the heavy metals. The study showed that Cd has the highest mobility in sludge treated soil followed by Cu, Cr, Zn, Ni and Pb.

Animals↗
PubMed1993

The role of stem cell factor in mobilization of peripheral blood progenitor cells: synergy with G-CSF.

The use of cytokine mobilized peripheral blood progenitor cells (PBPC) in transplantation following chemotherapy has led to enhanced engraftment. Granulocyte-colony stimulating factor (G-CSF) has been shown in a number of clinical studies to be an effective mobilizer of PBPC. Preclinical data in mice and primates have demonstrated a potential role for the use of stem cell factor (SCF) in mobilization of PBPC. In the studies presented here, low doses of SCF are shown to synergize with optimal doses of G-CSF to enhance the number and quality of PBPC compared to G-CSF alone. Phase I studies using r-metHuSCF demonstrated mast cell-related dose limiting effects. The data presented here have led to Phase I/II studies to evaluate the potential use of low doses of SCF in combination with G-CSF for mobilization of PBPC.

Animals↗
PubMed1998

Marrow repopulating cells in mobilized PBPC can be serially transplanted for up to five generations or be remobilized in PBPC reconstituted mice.

We have evaluated the durability of engraftment and the potential of remobilization in mice reconstituted with mobilized peripheral blood progenitor cells (PBPC). Female mice which had been reconstituted with cytokine-mobilized PBPC from male donors were serially transplanted into second, third, fourth and fifth lethally irradiated female recipients at intervals of 6-10 months. Male-derived hematopoiesis was determined in recipient mice at each serial transplantation. Male-positive CFCs were detected after 5 passages for 45 months, but declined from >95% at passage 1 to 74% at passage 2, 33% at passage 4, and 28% at passage 5. Long-term survival also declined from 97% at passage 2 to 53% at passage 4, and 27% at passage 5. The results demonstrated that mobilized PBPC were able to provide engraftment for more than 45 months, but the engraftment provided by mobilized PBPC decreased at each serial passage. In addition, mice reconstituted with mobilized PBPC (at 1 year post transplantation) were treated with the same cytokines as in the primary mobilization (remobilization). The remobilized PBPC were harvested and transplanted into lethally irradiated secondary recipients. Male-derived CFCs were evaluated at 20 months post transplantation. Mice transplanted with PBPC remobilized with rhG-CSF or rhG-CSF plus rrSCF-PEG showed 70% and 89% male-positive CFCs respectively, demonstrating that mice reconstituted with mobilized PBPC could be remobilized and that the remobilized PBPC were also capable of providing long-term hematopoietic reconstitution. Our studies demonstrated that mobilized PBPC have extensive proliferative or self-renewal capacity to provide durable engraftment and that marrow repopulating cells in PBPC reconstituted mice can be remobilized, suggesting that patients who relapse after PBPC transplantation may be remobilized for a second transplantation to support additional chemotherapy.

Animals↗
PubMed1985

Potential impact of acid precipitation on arsenic and selenium.

The potential impact of acidic precipitation on the environmental mobility of the metalloids arsenic (As) and selenium (Se) has not been given much attention and is poorly understood. As with other elements, the interest here is the potential effect of environmental acidification on environmental behavior in ways that are relevant to human exposure to these metalloids. Available information on acid precipitation and the environmental behavior of these metalloids do, however, permit some preliminary conclusions to be drawn. Both As and Se appear to be mobilized from household plumbing into tap water by the corrosive action of soft, mildly acidic water, while surface water catchment systems in areas impacted by acidic deposition may contain elevated soluble As levels. Acidification of aquatic ecosystems that are drinking water sources may pose the prospect of enhanced release of As from sediment to water as well as reduction in water levels of Se. Acidification of ground waters, where As appears to be especially mobile, is of particular concern in this regard. The potential impact of acidic deposition on As and Se in soils cannot readily be assessed with respect to human exposure, but it would appear that the behavior of these metalloids in poorly buffered, poorly immobilizing soils, e.g., sandy soils of low metal hydrous oxide content, would be most affected. The effect is opposite for the two elements; lowered pH would appear to enhance As mobility and to reduce Se availability. Altered acidity of both soil and aquatic systems poses a risk for altered biotransformation processes involving both As and Se, thereby affecting the relative amounts of different chemical forms varying in their toxicity to humans as well as influencing biogeochemical cycling.

Acids↗
PubMed2005

Electromagnetic field emitted by 902 MHz mobile phones shows no effects on children's cognitive function.

The present study investigated the potential effects of a standard 902 MHz global system for mobile communication (GSM) mobile phone on 10-14 years old children's cognitive function. A total of 32 children (16 boys, 16 girls) participated with their own and parental consent. The subjects were 10-14 years old (mean 12.1 years, SD 1.1). They performed a battery of cognitive tests twice in a counter-balanced order: once while exposed to an active mobile phone and once during exposure to an inactive phone. The tests were selected from those we used earlier with adults. The statistical analyses showed no significant differences between the mobile phone off and on conditions in reaction times and accuracy over all tests or in any single test. It was concluded that a standard mobile phone has no effect on children's cognitive function as measured by response speed and accuracy. The present results challenge some earlier findings suggesting that the electromagnetic field (EMF) created by an active mobile phone would facilitate cognitive functioning.

Adolescent↗
PubMed2004

Integrated optical-fiber capillary electrophoresis microchips with novel spin-on-glass surface modification.

This paper presents a novel micro-capillary electrophoresis (CE) chip with embedded optical fibers for the on-line detection of DNA samples. The optical fibers are pre-etched and then inserted directly into fiber channels incorporated within low-cost soda-lime glass substrates. The embedded optical fibers are precisely aligned with the microfluidic channels such that the induced fluorescence signals from labeled bio-samples can be detected. This arrangement avoids the requirement for delicate optical alignment procedures and equipment. Surface modification of the CE channels is accomplished by means of a simple and reliable organic-based spin-on-glass (SOG) method. The zeta potential distribution and the corresponding electroosmotic mobility of the fluid are simulated numerically for the modified and non-modified channel surfaces, and then both sets of results are verified experimentally. The present results indicate that the value of the zeta potential for a surface with an SOG coating is 19.3 times smaller than that of an untreated surface. A phiX-174 DNA marker fluid is used to evaluate the injection and separation performance of the developed micro-CE device. Furthermore, the long-term stability of the SOG-coated surface is also investigated. The experimental data reveal that the microchip device is capable of providing highly efficient separations of bio-molecules, and that the SOG layer retains its low zeta potential characteristics for at least 45 days. The present results confirm the effectiveness of the proposed micro-CE chip in performing the on-line detection of DNA samples, and indicate that the SOG process represents a simple and reliable solution for the surface modification of glass-based microchannels.

Bacteriophages↗
PubMed1993

Effect of recombinant human granulocyte colony-stimulating factor treatment of mice on spleen colony-forming unit number and self-renewal capacity.

The aim of this investigation was to find out whether the self-renewal capacity of spleen colony-forming units (CFU-S) changed after treatment of CBF1 (CBA-LacxC57B1/6) F1 female mice with granulocyte colony-stimulating factor (G-CSF). In addition, the possibility of synergism between erythropoietin (Epo) and G-CSF in blood stem cell mobilization was studied. Twenty-five and 250 micrograms/kg/d injections of recombinant human (rh) G-CSF expanded the number of CFU-S in peripheral blood four- and 32-fold. The self-maintenance potential of CFU-S did not change significantly. The treatment of mice with 250 micrograms/kg/d rhG-CSF resulted in a two-fold increase of spleen cellularity and a 15-fold augmentation of CFU-S number, without noticeable changes in their self-renewal capacity. Moderate changes in CFU-S number were observed after Epo administration in spleen and peripheral blood; however, no significant synergistic effect of Epo was detected with either dose of rhG-CSF. The multifold increase of CFU-S number in peripheral blood following rhG-CSF administration, with no reduction in their generating potential, suggests that rhG-CSF-mobilized blood stem cells provide an appropriate source for reconstitution of the hematopoietic system.

Animals↗
PubMed2003

Erythropoietin is a potent physiologic stimulus for endothelial progenitor cell mobilization.

Increasing evidence suggests that postnatal neovascularization involves the recruitment of circulating endothelial progenitor cells (EPCs). Hematopoietic and endothelial cell lineages share common progenitors. Cytokines formerly thought to be specific for the hematopoietic system have only recently been shown to affect several functions in endothelial cells. Accordingly, we investigated the stimulatory potential of erythropoietin (Epo) on EPC mobilization and neovascularization. The bone marrow of Epo-treated mice showed a significant increase in number and proliferation of stem and progenitor cells as well as in colony-forming units. The number of isolated EPCs and CD34+/flk-1+ precursor cells was significantly increased in spleen and peripheral blood of Epo-treated mice compared with phosphate-buffered saline-treated mice. In in vivo models of postnatal neovascularization, Epo significantly increased inflammation- and ischemia-induced neovascularization. The physiologic relevance of these findings was investigated in patients with coronary heart disease. In a multivariate regression model, serum levels of Epo and vascular endothelial growth factor were significantly associated with the number of stem and progenitor cells in the bone marrow as well as with the number and function of circulating EPCs. In conclusion, the present study suggests that Epo stimulates postnatal neovascularization at least in part by enhancing EPC mobilization from the bone marrow. Epo appears to physiologically regulate EPC mobilization in patients with ischemic heart disease. Thus, Epo serum levels may help in identifying patients with impaired EPC recruitment capacity.

Animals↗
PubMed2000

Electropherogram of capillary zone electrophoresis with effective mobility axis as a transverse axis and its analytical utility. I. Transformation applying the hypothetical electroosmotic flow.

For capillary zone electrophoresis, a new method of transformation from migration time to effective mobility was proposed, in which the mobility increase due to Joule heating and the relaxation effect of the potential gradient were eliminated successfully. The precision of the mobility evaluated by the proposed transformation was discussed in relation to the analysis of rare earth ions. By using the transformation, almost the same pherograms could be obtained even from the pherograms obtained originally at different applied voltages.

Animals↗