Improvement of dry float–sink separation of smaller sized spheres by reducing the fluidized bed height

In this study, the influence of the fluidized bed height on the float–sink of different sized spheres in a gas–solid fluidized bed was investigated. Fluidized beds with heights h = 200, 150, 100 and 50 mm were prepared using a cylindrical column of inner diameter = 290 mm and a mixture of zircon sand and iron powder as the fluidized medium. Float–sink experiments were carried out using density adjusted spheres of diameter D_sp = 40, 30, 20 and 10 mm. It was found that the float–sink performance at D_sp ?20 mm is not affected by the height of the bed, and the sharpness of separation (the density range where spheres neither float nor sink completely) is less than or equal to 200 kg/m³. In the case of D_sp = 10 mm, the sharpness of separation is a larger value (1100 kg/m³ at h = 200 mm), whereas it decreases with decreasing h and is 400 kg/m³ at h = 50 mm. The fluctuation of the surface height of the fluidized bed was visually recorded. The fluctuation is reduced by reducing h. The fluctuation vs. h correlates with the sharpness of separation at D_sp = 10 mm vs. h. These results indicate that the dry float–sink separation of smaller sized spheres is improved as the fluctuation of fluidized bed surface is decreased by reducing the fluidized bed height.     

Augmented proliferation of human alveolar macrophages after allogeneic bone marrow transplantation

After allogeneic bone marrow transplantation (allo-BMT), recipient alveolar macrophages (AM) are gradually replaced by AM of the donor origin. An influx of mononuclear phagocytes of donor origin to the lung is responsible for the repopulation, but the detailed kinetics remain unclear. We therefore studied 24 BMT recipients who underwent bronchoalveolar lavage (BAL) from 24 to 83 days after BMT. AM cell number, size, morphology, proliferating ability, and genotype of AM were measured. Before day 50, the number and size of AM in BAL fluid were similar to those of normal nonsmokers. However, after day 50, the mean number of AM increased threefold and the mean cell size decreased due to the increase of small AM. These small cells are presumably of donor origin based on DNA fingerprinting analysis and based on fluorescence in situ hybridization for the Y chromosome in a sex-mismatched case. Immunohistochemistry and cell cycle analysis demonstrated that the increase in AM number coincided with a remarkable increase of AM expressing proliferating cell nuclear antigen, suggesting that small AM are proliferating. This is the first report representing that augmented proliferation of donor AM in situ may contribute to the reconstitution of AM population after BMT.     

Thrombopoietin and erythropoietin activate inside-out signaling of integrin and enhance adhesion to immobilized fibronectin in human growth-factor-dependent hematopoietic cells

Erythropoietin (EPO) and thrombopoietin (c-MPL ligand; TPO) are structurally similar cytokines and support respectively, the proliferation and differentiation for erythroid and megakaryocytic lineages, as well as more primitive progenitors. We studied the effect of these cytokines on the induction of adhesion of human growth-factor-dependent hematopoietic cells to immobilized fibronectin, which is a main component of the extracellular matrix in the bone marrow. MO7ER cells that are genetically engineered to express human EPO receptor and MO7e cells that express endogenous c-MPL were used. Stimulation with either TPO or EPO induced rapid increases in adhesion of M07ER cells to fibronectin without apparent change of expression of integrins. Experiments with inhibitory monoclonal antibodies (mAbs) demonstrated that CD41, which has been reported to be involved in TPO-induced adhesion of megakaryocytic cells, is not responsible for this enhanced adhesion. Anti-beta 1 integrin mAb inhibited adhesion completely, while inhibition by anti-alpha 4 integrin mAb and anti-alpha 5 integrin mAb was partial. Combination of anti-alpha 4 mAb plus anti-alpha 5 mAb completely abolished adhesion, as did anti-beta 1 mAb, suggesting that the adhesion is mediated by both alpha 4 beta 1 and alpha 5 beta 1 integrins. Experiments using inhibitors suggested that ligand binding followed by activation of intracellular tyrosine kinases along with PI3-kinase activation is required. After stimulation of M07ER cells with either TPO or EPO, fibronectin-attached cells, but not cells in suspension, showed tyrosine phosphorylation of focal adhesion kinase, which plays a central role in integrin-mediated signaling. These data suggest that TPO and EPO might be involved in homing/migration to the bone marrow microenvironment by hematopoietic cells that express corresponding receptors.     

Effects of parent Shamo cocks on the histochemical properties of M. iliotibialis lateralis and M. supracoracoideus on their crossbred broilers

Histochemical studies of mammary gland sections obtained from healthy lactating Prim' Holstein cows contained numerous corpora amylacea, mainly located in active alveoli. Observations by electron microscopy revealed a cauliflower shape, with a fibrillar or multilayered organization. Mineral studies confirmed the presence of high calcium levels (12.3% of dry matter) and phosphorus (7.4%) in the corpora amylacea composition. These bodies stained positive to Von Kossa silver nitrate and to Periodic acid-Schiff. However, depending on the gland of origin, corpora amylacea stained positive or negative to Congo red. Histochemical studies seemed, therefore, insufficient to determine the presence or absence of amyloid. The amount of total protein varied by approximately 25%. Immunoblotting and analysis of the amino acid sequence of a peptidic fragment obtained from corpora amylacea gave clear evidence of the occurrence of caseins, beta-lactoglobulin and alpha-lactalbumin. However, the comparison between the amino acid composition of corpora amylacea and those of the main milk proteins indicated the presence of other proteins. Electrophoretic analysis also gave evidence of the presence of several other proteins, i.e. glycoproteins. Therefore, it is probable that corpora amylacea composition is much more complex than previously reported.     

Transarterial immuno-embolization therapy in patients with hepatocellular carcinoma

Surgical resection, transcatheter arterial embolization (TAE) and percutaneous ethanol injection therapy (PEIT) are effective for hepatocellular carcinoma (HCC), but the recurrence rate is high. We have devised a new therapy of transarterial immuno-embolization (TIE) with OK-432, fibrinogen and thrombin, and 2 cases are reported. Case 1: A 78-year-old Japanese male with HCC (diameter, 4 cm in subsegment 5) received TIE. The tumor size was markedly decreased, and the patient survived for more than 3 years without recurrence. Case 2: A 61-year-old Japanese male with HCC (diameter, 4.5 cm in segment 5) received hepatic subsegmentectomy following TIE. Histological examination of resected specimens following TIE showed massive infiltration of mononuclear cells in the main tumor. Tumor recurrence had developed three times thereafter, but was effectively treated by TIE. TIE may be an effective therapy for HCC.     

A research program for dynamic fracture evaluation of Japanese carbon steel pipes

A research program was developed to investigate the dynamic load effect on fracture behavior of Japanese carbon steel STS410 pipe. The program comprises material tests, pipe fracture tests and development of estimation scheme. Material property tests showed that the flow stress was nearly constant or slightly increased with strain rate. Pipe tests showed that fracture load was nearly predicted by the net-section collapse criterion for both quasi-static and dynamic loading. Significant dynamic effect was not observed for STS410 carbon steel piping. Crack growth was well formulated by using J-integral parameter for low cycle fatigue with large scale yielding. Combining the crack growth behavior and unstable fracture criterion, an estimation scheme was newly developed and validated for constant amplitude cyclic loading conditions.     

Localization and in vitro mutagenesis of the active site in the Saccharomyces cerevisiae mRNA capping enzyme

The yeast mRNA capping enzyme is composed of 52 (alpha) and 80 kDa (beta) polypeptides, which are responsible for its mRNA guanylyltransferase and RNA 5'-triphosphatase activities, respectively. We isolated the gene encoding the alpha subunit (CEG1) and showed that CEG1 is essential for yeast cell growth [Shibagaki et al., (1992) J. Biol. Chem. 267, 9521-9528]. In this study, CEG1 was expressed in Escherichia coli and the alpha subunit protein was purified to near homogeneity. A [32P]GMP-bound tryptic peptide derived from the recombinant enzyme-[32P]GMP covalent reaction intermediate was converted to a [32P]phosphoryl-peptide through periodate oxidation followed by beta-elimination. Hydrolysis of the [32P]phosphoryl-peptide with alkali resulted in [32P]N epsilon-phospholysine as the only phosphoamino acid, indicating that GMP in the enzyme-GMP complex is bound to a lysine residue via a phosphoamide linkage. Microsequencing of the [32P]GMP-peptide showed that the GMP binding site was located in the region between amino acids 60 and 75, which contained an internal trypsin-resistant lysine at position 70. CEG1 was subjected to site-directed mutagenesis and the mutant proteins were expressed in E. coli. Substitution of His or Ile for Lys70 entirely abolished the enzyme-GMP formation activity, and this mutation was lethal to yeast in vivo, supporting the notion that the active site in the alpha subunit is located at Lys70. Replacement of Lys70 with Arg reduced the ability to form the enzyme-GMP complex; however, yeast cells bearing this allele were not viable. A series of mutations, including 8 amino acid replacements and 3 insertions, near the active site (Lys70-Thr-Asp-Gly motif) were also introduced and the mutant polypeptides were examined for catalytic activity in vitro as well as yeast cell viability in vivo. There was a good correlation between the in vitro and in vivo functions of the mutant proteins, except when Asp72 was replaced with Glu, which allowed formation of the enzyme-GMP complex but failed to support cell growth. The results with Lys70 to Arg and Asp72 to Glu substitutions indicated that guanylyltransfer to RNA and/or additional roles besides cap formation per se are impaired in these mutant proteins.     

Cellular conditions for intramuscular fat deposition in Japanese Black and Holstein steers

The experiment was conducted to study the development of intramuscular fat in Japanese Black (JB) compared to Holstein (HS) steers and to find breed differences for fat depot development and distribution in the carcass under equal feeding conditions. Additional to slaughter samples, biopsy samples of longissimus muscle (LM) and subcutaneous fat, taken at 10, 14, 18, and 22 months of age, were used for histological and molecular investigations. Japanese Black steers stored about 14% more fat in the LM (P = 0.001), resulting in larger marbling flecks (P < 0.001). Muscle fibers and intramuscular adipocytes in both breeds responded to the high energy feeding with significant enlargement, which was faster in JB. Histograms of intramuscular adipocytes size showed a shift toward larger cells during growth, but also the abundance of small, developing adipocytes. This development was accompanied by a correlated up-regulation of adipogenic genes until 22 months of age.