Prevalence and growth characteristics of malignant stem cells in B-lineage acute lymphoblastic leukemia

We used a stroma-supported culture method to study the prevalence and growth characteristics of malignant stem cells in acute lymphoblastic leukemia (ALL). In 51 of 108 B-lineage ALL samples, bone marrow-derived stroma not only inhibited apoptosis of ALL cells but also supported their proliferation in serum-free medium. When single leukemic cells were placed in the stroma-coated wells of microtiter plates, the percentage of wells with leukemic cell growth after 2 to 5 months of culture ranged from 6% to 20% (median, 15%; 5 experiments). The immunophenotypes and genetic features of cells recovered from these cultures were identical to those noted before culture. All cells maintained their stroma dependency and self-renewal capacity. Leukemic clones derived from single cells contained approximately 10(3) to 10(6) cells after 1 month of culture; other clones became detectable only after prolonged culture. Cell growth in stroma-coated wells correlated with the number of initially seeded cells (1 or 10; r = .87). However, the observed percentages of positive wells seeded with 10 cells always exceeded values predicted from results with single-cell-initiated cultures (P < .003 by paired t-test), suggesting stimulation of leukemic cell growth by paracrine factors. In conclusion, the proportion of ALL cells with clonogenic potential may be considerably higher than previously thought.     

Enhancement of ectopic bone formation in mice with a deficit in Fas-mediated apoptosis

Bone formation is under the control of cytokines as well as growth factors such as bone morphogenetic proteins (BMP). This suggests the possibility that osteogenesis might be modulated by factors which also modulate the immune system. To test whether immune disorders in the host may influence bone formation, we studied BMP-induced bone formation in a C3H/HeJ strain of mice bearing a mutant gene, the lymphoproliferation gene (lpr) or the generalized lymphoproliferative disease gene (gld), both of which are known to be a Fas deletion mutant and a Fas ligand mutant, respectively, and to induce immune disorders via a deficit in Fas-mediated apoptosis. Crude BMP derived from bovine bone were injected into the muscular tissue in the femur of adult C3H/HeJ mice or C3H/HeJ mice bearing an lpr or gld gene. Quantitative analysis of the resulting ectopic bone formation by X-ray photography 2 weeks after injection revealed that the presence of either the lpr or gld gene caused a bone mass significantly larger in dimension than that seen in the wild type mice. Histological examination also revealed the different influence between these mutant genes on the level of bone formation exhibited by hyaline cartilage and bone trabeculae. Based on these results, we discussed the possible mechanisms of the enhanced ectopic bone formation under the deficit in Fas-mediated apoptosis.     

Needle penetrometry with variable force loading for measuring viscosity of pelletized coal upon heating

A needle penetrometry was performed loading steady force in a range from 1×10⁻³ to 2 N to pelletized coal upon heating via a cylindrical needle. From the observed effects of shear rate on apparent viscosity of softening coal pellet, defined as the shear-rate to shear-stress ratio, it was found that the pellet behaved as a Newtonian fluid for shear rates lower than a critical one while as a pseudo-plastic fluid for higher shear rates. The penetrometry was also carried out varying the force with time. The variable force loading enabled to maintain the shear rate well below the critical one, and thereby to measure the apparent viscosity of coal pellets as Newtonian fluids over a temperature range from 600 to 800 K. Upon heating at 10 K min⁻¹, the apparent viscosity of Goonyella coal pellet decreased from about 10¹⁰ Pa s at 640 K down to a minimum of about 10⁴ Pa s at 755 K, and increased up to 10⁹ Pa s at 800 K. In a course of heating as above, the viscosity of Blind Canyon coal pellet decreased above 600 K, underwent a minimum of about 10⁶ Pa s at 715 K, and increased up to 10¹⁰ Pa s at 770 K. Decreasing the heating rate from 10 to 3 K min⁻¹ caused the minimum viscosities of the pellets to increase by 1-2 orders of magnitude.     

Cloning and nucleotide sequence of the beta-D-glucosidase gene from Bifidobacterium breve clb, and expression of beta-D-glucosidase activity in Escherichia coli

Genomic DNA encoding a beta-D-glucosidase (EC 3.2.1.21), which has beta-D-fucosidase activity, was cloned from Bifidobacterium breve clb. We sequenced a 1.9-kbp cloned DNA fragment that contained a single open reading frame encoding 460 amino acids with a calculated molecular mass of 51,513 Da. A putative ribosome binding site was found 5 bp upstream of the initiation codon. The amino acid sequence of this beta-D-glucosidase from Bifidobacterium breve clb had 46% identity with that of beta-glucosidase from Microbispore bispore. The enzyme of Bifidobacterium breve clb was expressed in Escherichia coli. A cell-free extract prepared from the recombinant strain showed 80 to 90-fold more beta-D-glucosidase activity than that from Bifidobacterium breve clb. The recombinant enzyme was purified to homogeneity from cell-free extracts of the recombinant strain using 4 column chromatographies. The recovery of enzyme from the recombinant strain was about 138-fold-higher than that of Bifidobacterium breve clb. The enzymatic properties were similar to those of Bifidobacterium breve clb. For application of this recombinant enzyme, we attempted to synthesize a disaccharide that seemed to be specifically assimilated by Bifidobacteria using the condensation activity of the enzyme.     

Involvement of superoxide and nitric oxide on airway inflammation and hyperresponsiveness induced by diesel exhaust particles in mice

We previously demonstrated that chronic intratracheal instillation of diesel exhaust particles (DEP) induces airway inflammation and hyperresponsiveness in the mouse, and that these effects were partially reversed by the administration of superoxide dismutase (SOD). In the present study, we have investigated the involvement of superoxide in DEP-induced airway response by analyzing the localization and activity of two enzymes: (1) a superoxide producer, NADPH cytochrome P-450 reductase (P-450 reductase), and (2) a superoxide scavenger, SOD, in the lungs of the exposed mice and controls. P-450 reductase was detected mainly in ciliated cells and clara cells: its activity was increased by the repeated intratracheal instillation of DEP. While CuZn-SOD and Mn-SOD were also present in the airway epithelium, their activity was significantly decreased following DEP instillation. Exposure to DEP doubled the level of nitric oxide (NO) in the exhaled air. DEP exposure also increased the level of constitutive NO synthase (cNOS) in the airway epithelium and inducible NO synthase (iNOS) in the macrophages. Pretreatment with N-G-monomethyl L-arginine, a nonspecific inhibitor of NO synthase, significantly reduced the airway hyperresponsiveness induced by DEP. These results indicate that superoxide and NO may each contribute to the airway inflammation and hyperresponsiveness induced by the repeated intratracheal instillation of DEP in mice.     

Charge—discharge characteristics and structural change in various niobium sulfide cathodes for lithium-nonaqueous secondary batteries

Electrochemical behaviors of niobium sulfides such as 3R-Nb_1+xS₂ (x = 0.00 ≈ 0.25), 2H-NbS₂ and NbS₃ for lithium batteries are investigated in 1 M LiClO₄-propylene carbonate solution. The cathodic reaction in the charge-discharge cycle of the disulfides is shown as follows:

X-ray diffraction measurement and open circuit potential data show that 3R-Nb_1.00S₂ gives continuous structural change along c-axis reversibly with cycling, where x takes from 0 to 1, whereas 2H-NbS₂ gives the stepwise change also reversibly with cycling, where x is at least 1. On the other hand, NbS₃ yields the ternary phase of Li_xNbS₃ giving an amorphous X-ray pattern and shows irreversible structural change with cycling. When the charge-discharge cyclings are carried out at a constant capacity of 0.50 equivalent mole^?1 of sulfide and a current density of 0.20 mA cm^?2, 3R-Nb_1.00S₂ can be cycled over 100 times without an increase in the charge potential, whereas in 2H-NbS₂ and NbS₃ the charge potentials rise to 4.6 V vs Li/Li⁺ with cycling, which limits the cycle number to 30 ≈ 50 times. This would be due to the fact that 3R-Nb_1.00S₂ shows the reversible structural change with keeping the original crystal lattice with cycling.     

Intake of different eicosapentaenoic acid-containing lipids and fatty acid pattern of plasma lipids in the rats

The ethyl ester of eicosapentaenoic acid (EPA) is the only pure EPA-containing lipid available in bulk for oral administration. However, there is doubt as to whether EPA ethyl ester can efficiently increase the plasma levels of EPA in comparison with the ability of other kinds of EPA-containing lipids to do so. Therefore, two other kinds of EPA-containing lipids were prepared to study the efficiency of oral administration of those lipids for increasing the EPA content in plasma phospholipids and cholesteryl esters. EPA-containing lipids which were investigated were [A], 1,2,3-trieicosapentaenoyl-glycerol, [B] 2-eicosapentaenoyl-phosphatidylcholine and [C] ethyl ester of EPA. An adjusted amount of lipids [A], [B] and [C] was administered to rats through a gastric tube for 4 days (the first experiment) or for 10 days (the second experiment), and the fatty acid composition of plasma phospholipids and cholesteryl esters was determined. In the first experiment, there were no significant differences in the efficiency for increasing EPA levels in either phospholipids or cholesteryl esters among the lipids. In the second experiment, the EPA levels of both plasma phospholipids and cholesteryl esters of rats administered ethyl ester of EPA were significantly higher than those of rats administered 2-eicosapentaenoyl-phosphatidylcholine. The EPA levels of the rats administered 1,2,3-trieicosapentaenoylglycerol were between the levels of the two groups mentioned above, but the differences in the EPA levels were not significant. Although an ethyl ester-type molecule is not a naturally occurring lipid, ethyl ester of EPA is equal to 1,2,3-trieicosapentaenoyl-glycerol and appears to be superior to 2-eicosapentaenoyl-phosphatidylcholine as to the efficiency for increasing EPA levels in total plasma phospholipids and plasma cholesteryl esters.     

Time optimal discrete control system with bounded state variable

This paper considers the minimum time regulator problem for a linear time-invariant discrete system with input and state variable constraints. The state space techniques are used throughout. Conditions for controllability of the system are examined in detail and an algorithm for generating optimal controls is given such that computor implementation is possible for higher-order systems.