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Anti-vascular endothelial growth factor (VEGF) therapy has demonstrated efficacy in treating human metastatic cancers, but therapeutic resistance is a practical limitation and most tumors eventually become unresponsive. To identify microenvironmental factors underlying the resistance of cancer to antiangiogenesis therapy, we conducted genomic analyses of intraperitoneal ovarian tumors in which adaptive resistance to anti-VEGF therapy (B20 antibody) developed. We found that expression of the microseminoprotein, prostate-associated (MSMP) gene was substantially upregulated in resistant compared with control tumors. MSMP secretion from cancer cells was induced by hypoxia, triggering MAPK signaling in endothelial cells to promote tube formation in vitro. Recruitment of the transcriptional repressor CCCTC-binding factor (CTCF) to the MSMP enhancer region was decreased by histone acetylation under hypoxic conditions in cancer cells. MSMP siRNA, delivered in vivo using the DOPC nanoliposomes, restored tumor sensitivity to anti-VEGF therapy. In ovarian cancer patients treated with bevacizumab, serum MSMP concentration increased significantly only in non-responders. These findings imply that MSMP inhibition combined with the use of antiangiogenesis drugs may be a new strategy to overcome resistance to antiangiogenesis therapy.

AbstractThe energy consumption and indoor temperature of 13 housing units sited in the northern region of Honshu Island, Japan were investigated for a full two years from Dec. 2002 to Nov. 2004. Five of the houses were installed with all-electric equipment. The annual and daily energy consumption profiles of two typical houses were analyzed, and the annual energy consumption per house was found to range from 40GJ/year to 120GJ/year. The difference of energy consumption between the two years was very small, while in some houses, energy consumption during the second year decreased due to a rise in energy saving awareness. The results indicated that the characteristics of energy consumption were not only greatly influenced by regional climate but also by the use of household equipment and lifestyle. Finally, the relationship between energy consumption for space heating and indoor temperature is discussed.

Plasmodium falciparum causes the most severe form of malaria. Utilization of fatty acids in serum is thought to be necessary for survival of this parasite in erythrocytes, and thus characterization of the parasite fatty acid metabolism is important in developing a new strategy for controlling malaria. Here, we examined which combinations of fatty acids present in human serum support the continuous culture of P. falciparum in serum-free medium. Metabolic labelling and gas chromatography analyses revealed that, despite the need for particular fatty acids for the growth of intraerythrocytic P. falciparum, it can metabolize a broad range of serum-derived fatty acids into the major lipid species of their membranes and lipid bodies. In addition, these analyses showed that the parasite's overall fatty acid composition reflects that of the medium, although the parasite has a limited capacity to desaturate and elongate serum-derived fatty acids. These results indicate that the Plasmodium parasite is distinct from most cells, which maintain their fatty acid composition by coordinating de novo biosynthesis, scavenging, and modification (desaturation and elongation).

In order to improve the durability of Pt coated Ti electrodes as an anode of the sulfuric acid system electrolysis, tantalum oxide (Ta2O5) additions have been used widely. However that is not enough for uses under severe conditions. In this study,[removed info]boric acid (H3BO3) was added into the Pt coating during manufacturing process of Ti/Pt-coated electrodes, and the (Pt + B2O3)/Ti electrodes were obtained. The effect of H3BO3 addition on the microstructure, surface area, and lifetime of (Pt + B2O3)/Ti electrodes was also investigated in detail. Commercially available titanium plates (10[removed info] 10[removed info]0.5mm) were employed for prepare Ti/ (Pt + B2O3) electrodes, where it was confirmed by XRD that B2O3 formed by heating the boric acid was amorphous in the temperature range of 250 to 550[removed info]. The coating solution of 2 μl of H2PtCl6[removed info]6H2O (Pt:50 g/l) dissolved in butanol and 5~20 mass% H3BO3 mixture was spread over the etched Ti plates. After drying at 70[removed info] for 30 min, the substances were heated at 250 to 550[removed info] for 10 min. The lifetime of the electrodes was examined by a direct current of 1 A using Pt plate as the counter electrode in 1 M H2SO4 solution at 40[removed info]. The end of life was determined as the time when the cell voltage changed 2 times of the beginning.

The study was designed to evaluate the effects of a traditional Chinese herbal medicine Hochu-ekki-to (Bu-zong-yi-qi-tang), which was composed of 10 herbal medicines and had been used for the treatment of oligospermia and as a postoperative medication in Japan, on bone loss in rats treated with a gonadotropin-releasing hormone (GnRH) agonist. Female rats at 40 weeks of age were divided into 4 groups of 8 rats each. In the three experimental groups, each animal received subcutaneous injections of the long-acting GnRH agonist, buserelin acetate, once every four weeks throughout the experiment. Beginning at 48 weeks of age, the experimental groups were given diets containing conjugated estrogens or Hochu-ekki-to for 8 weeks. The administration of the GnRH agonist reduced the bone mineral density in the whole femur to 91.0% of that in the control group. However, administration of conjugated estrogens and Hochu-ekki-to increased the serum concentrations of estradiol 16.8- and 5.3-fold respectively compared with concentrations in the GnRH agonist-treated group, resulting in the augmentation of the bone mineral density to 110.3% and 106.2% respectively. These findings indicate that Hochu-ekki-to enhances the reduced bone mineral density and causes a slight elevation of the serum estradiol levels in the chemically castrated rats.

We have measured the yield of the sputtered ions from the surface of solid targets of conductive materials (Al, Ni, Cu, Ag, Si) bombarded by Xeq+(q=15–44) at 1.0 MeV (vp=0.55 a.u). The experiment was performed using micro-bunched highly charged Xe ion beams from CRYEBIS at Kansas State University. The secondary ions were detected and mass analyzed by a TOF spectrometer. The secondary ion yields showed a significant increase with increasing projectile charge state q, demonstrating the electronic sputtering in the interaction of slow highly charged ions with conductive materials. The increase rate became larger above q=26 compared to lower charge states, and showed a more prominent increase above q=35. These strong onsets of the increase rate at particular incident charge states can be attributed to the opening up of new inner shell holes in the incident highly charged ions. This is the first systematic observation of electronic sputtering of conductive materials using slow highly charged heavy ions bearing high potential energy.

Abstract.: We investigated the effects of conjugated estrogens as an add-back replacement drug, incadronate sodium as a bisphosphonate, and alfacalcidol as a vitamin D3 analog on femoral bone mineral density (BMD) and bone mineral content (BMC) in female rats chronically treated with the gonadotropin-releasing hormone (GnRH) agonist leuprorelin acetate. The chemical castration of the rats by the administration of GnRH agonist for 16 weeks reduced the BMD values to 92.3%, 91.3%, and 93.3% of those of the normal control animals in the whole femur, metaphysis, and diaphysis of the femur, respectively. The BMC value was decreased to 91.0% of that of the normal control animals by the chronic GnRH agonist treatment. However, a simultaneous 8-week administration of conjugated estrogens, bisphosphonate, and vitamin D3 analog markedly augmented the BMC values to 110.3%, 110.1%, and 114.4%, respectively, of those in the rats treated with the GnRH agonist alone. These findings indicate that antiosteoporotic agents could be useful for preventing induced osteopenia under the careful monitoring of biochemical markers of osteoblastic activity or bone resorption and BMD or BMC in patients undergoing GnRH treatment.

New cubic leucite-type compounds, CsMSi2O6 (M=B0.2Al0.8,Al0.2Fe0.8), Cs2MSi5O12 (M=Cd, Mg, Ni, Zn) have been synthesized by the two-stage heat treatment of the solid-state reaction. The thermal expansion properties of the synthesized leucite-type compounds have been studied with HTXRD and LTXRD in the temperature range of 123 to 1273 K. The thermal expansion rate of CsB0.2Al0.8Si2O6 was found to be considerably smaller than that of CsAlSi2O6, while the thermal expansion property of Cs2MSi5O12 (M=Mg, Zn, Cd) was found to have a linear relationship in the temperature range of 298 to 1273 K. By using Rietveld analysis it was found that the thermal expansion rate decreased with increasing the Si—O—M(Si) angle for cubic leucite-type compounds at 298 K, and that the phase transitions of CsAlSi2O6 and Cs0.9Al0.9Si2.1O6 were due to the relationship between the bond angle of Si—O—M(Si) of the three-dimensional framework structure and the space ratio in the unit cell at 298 K.

The serine repeat antigen (SERA) is a vaccine candidate antigen of Plasmodium falciparum. Immunization of mice with Escherichia coli-produced recombinant protein of the SERA N-terminal domain (SE47') induced an antiserum that was inhibitory to parasite growth in vitro. Affinity-purified mouse antibodies specific to the recombinant protein inhibited parasite growth between the schizont and ring stages but not between the ring and schizont stages. When Percoll-purified schizonts were cultured with the affinity-purified SE47'-specific antibodies, schizonts and merozoites were agglutinated. Indirect-immunofluorescence assays with unfixed parasite cells showed that SE47'-specific immunoglobulin G (IgG) bound to SERA molecules on rupturing schizonts and merozoites but the IgG did not react with the schizont-infected erythrocytes (RBC). Furthermore, double-fluorescence staining against SE47'-specific IgG and anti-human RBC membrane IgG showed that the RBC membrane disappeared from SE47'-specific-IgG-bound schizonts after cultivation. These observations suggest that the SE47'-specific antibodies inhibit parasite growth by cross-linking SERA molecules that are associated with merozoites in rupturing schizonts with partly broken RBC and parasitophorous vacuole membranes, blocking merozoite release.

DRAP27, the monkey homolog of human CD9 antigen (DRAP27/CD9) and diphtheria toxin receptor (DTR) were expressed in mouse L cells. L cells transfected transiently with both DRAP27/CD9 and DTR cDNA bound approximately 10 times more diphtheria toxin (DT) than cells transfected with DTR alone. Stable L cell transfectants expressing both DTR and DRAP27/CD9 (LCH‐1 cells) had 15 times more cell surface DT‐binding sites and were 20 times more sensitive to DT than were stable L cell transfectants expressing DTR alone (LH‐1 cells). Increased DT‐binding and DT sensitivity were not due to increased DTR transcription or increased cell surface DTR protein. Co‐immunoprecipitation of DRAP27/CD9 with DTR and chemical cross‐linking suggest a tight association of these membrane‐bound proteins. In addition, the identity of DTR and a growth factor (HB‐EGF) was established. Immobilized DT specifically adsorbed HB‐EGF precursor solubilized from transfected L cells and [125I]DT bound to immobilized recombinant HB‐EGF. We conclude that DRAP27/CD9 associates tightly with DTR/HB‐EGF and up‐regulates the number of functional DTRs and DT sensitivity, and that HB‐EGF is identical to DTR.

Variable-energy (0–30 keV) positrons were used to study the depth distribution of heavy-ion induced vacancy type defects in the following specimens, SUS304 and SUS316 austenitic stainless steels, a SUS444 ferritic stainless steel and nickel metal, which were irradiated by 0.5 MeV He+, 2.0 MeV C+, 3.5 MeV Si2+ and 4.0 MeV Ni2+ ions up to 0.01 dpa at peak. Vacancy type effective residual defects (ERD) were evaluated from the line shape parameterS of Doppler broadened positron annihilation photon spectra. With increasing primary knockon atom (PKA) energy, a decrease of the vacancy type ERD was observed. The ERD differences among the specimens are discussed in comparison with theoretical predictions.

The formation mechanism of ZrSiO4 powders from ZrO2 and colloidal SiO2 sols has been studied. The change in the ZrSiO4 yield rate with seeding and non-seeding has also been investigated. The induction period of the ZrSiO4 yield-rate curve corresponded to the n=1 region, a simple nucleation region, on the Avrami plot. The n=1 region was a rather wide range at 1200–1300°C and the nucleation reaction was promoted in this temperature range. It was found that the ZrSiO4 yield rate became high, even if seeding was not used, when the heating rate was controlled at less than 1 °C min-1.

Nickel single crystal specimens were bombarded with various heavy ions, 0.2 MeV He+, 2.0 MeV C+, 3.5 MeV Si2+, 4.0 MeV Ni2+ ions, up to the same amounts of displacement per atom (dpa), and residual rate of defects in heavy-ion induced cascades was investigated by Rutherford backscattering (RBS) spectroscopy using channeling technique. Displacement damages are found to be variable with primary knock-on atom (PKA) energies, revealing a cascade quenching due to the recombination of point defects produced primarily. Residual rate of defects obtained experimentally are qualitatively in agreement with existing theoretical calculations.

Diphtheria toxin (DT) binds to the epidermal growth factor (EGF)-like domain of human membrane-anchored heparin-binding EGF-like growth factor (proHB-EGF), the human DT receptor (DTR). DT does not bind to mouse proHB-EGF because of amino acid substitutions within the EGF-like domain. We made 10 independent mutants, replacing a single amino acid within the EGF-like domain of human DTR/proHB-EGF with the corresponding amino acid residue in mouse proHB-EGF. The mutant proteins were transiently expressed in mouse L cells either expressing or not expressing DRAP27/CD9, and DT binding was measured. DT binding activity of GST fusion proteins containing the mutated EGF-like domain was also determined by a cell-free binding assay. The largest effect was seen with E141H, and second largest effects were seen with F115Y and L127F in all of the assay systems. We conclude that Phe115, Leu127, and Glu141 are critical amino acid residues for DT binding. A computer model of the tertiary structure of the EGF-like domain of human DTR/proHB-EGF was made. The model predicts that three amino acid residues critical for DT binding activity, Phe115, Leu127, and Glu141, are all located on the same face of the EGF-like domain, suggesting that this face of DTR/proHB-EGF interacts with the receptor-binding domain of DT.

Human heparin-binding epidermal growth factor (EGF)-like growth factor (HB-EGF) expressed on Chinese hamster ovary (CHO) cells is synthesized as a 19-kDa major, and 22- and 27-kDa minor, membrane-anchored precursors (proHB-EGF). In contrast, the 27-kDa species is major and the 19- and 22-kDa ones are minor in mouse proHB-EGF. The juxtacrine growth factor activities of human and mouse proHB-EGFs on CHO cells toward EP170.7 cells in co-culture are significantly different. To investigate the relationship between the juxtacrine growth factor activities and the molecular species, we prepared human-mouse chimeras. Chimeras that have the human amino-terminal sequence with a mouse EGF-like domain showed approximately 8-fold up-regulation of the juxtacrine growth factor activity and the predominance of a 19-22-kDa major species. In contrast, chimeras that have the mouse amino-terminal sequence with a human EGF-like domain showed approximately 5-fold down-regulation of the juxtacrine activity and the predominance of the 27-kDa major species. A Gly32.HB-EGF (117-amino acid form), which is amino-terminally extended, induced the same mitogenic activity as that of Arg73.HB-EGF (75-amino acid form), which is amino-terminally truncated. These results strongly suggested that amino-terminal processing of human proHB-EGF would be required for up-regulation of its juxtacrine growth factor activity, but not for its paracrine activity.

Bacillus megaterium is known to have several genes that code for isozymes of glucose dehydrogenase. Two of them, gdhI and gdhII, were cloned from B. megaterium IAM1030 in our previous work (T. Mitamura, R. V. Evora, T. Nakai, Y. Makino, S. Negoro, I. Urabe, and H. Okada, J. Ferment. Bioeng. 70:363-369, 1990). In the present study, two new genes, gdhIII and gdhIV, were isolated from the same strain and their nucleotide sequences were identified. Each gene has an open reading frame of 783 bp available to encode a peptide of 261 amino acids. Thus, a total of four glucose dehydrogenase genes have been cloned from B. megaterium IAM1030. In addition, this strain does not seem to have other glucose dehydrogenase genes that can be distinguished from the four cloned genes so far examined by Southern hybridization analysis. The two newly cloned genes were expressed in Escherichia coli cells, and the products, GlcDH-III and GlcDH-IV, were purified and characterized and compared with the other isozymes, GlcDH-I and GlcDH-II, encoded by gdhI and gdhII, respectively. These isozymes showed different mobilities in sodium dodecyl sulfate-polyacrylamide gel electrophoresis (GlcDH-I greater than GlcDH-III = GlcDH-IV greater than GlcDH-II), although they have the same number of amino acid residues. Double-immunodiffusion tests showed that GlcDH-I is immunologically different from the other isozymes and that GlcDH-III and GlcDH-IV are identical to one another but a little different from GlcDH-II. These glucose dehydrogenases were stabilized in the presence of 2 M NaCl. The effect of NaCl was especially large for GlcDH-III, which is most unstable enzyme. Kinetic studies showed that these isozymes are divided into two groups with respect to coenzyme specificity, although they can utilize both NAD and NADP: GlcDH-III and GlcDH-IV prefer NAD, and GlcDH-I and GlcDH-II prefer NADP. The phylogenic relationship of these glucose dehydrogenase genes is also discussed.

The membrane anchored form of human heparin-binding epidermal growth factor-like growth factor (HB-EGF) acts as the diphtheria toxin (DT) receptor. Transfection of human HB-EGF cDNA into mouse LC cells, L cells stably expressing DRAP27, conferred sensitivity to DT, but transfection of mouse HB-EGF cDNA did not. To define the essential regions of HB-EGF that serve as the functional DT receptor, we examined the sensitivity to DT and DT binding of cells expressing several human/mouse HB-EGF chimeras. It was found that DT binds to the EGF-like domain of the human HB-EGF. However, mouse HB-EGF does not serve as a functional DT receptor due to non-conserved amino acid substitutions in this domain. In addition, CRM197, a non-toxic mutant of DT, inhibited strongly the mitogenic activity of the secreted form of human HB-EGF, but not of mouse HB-EGF and other EGF receptor-binding growth factors. These results confirmed further that DT interacts with the EGF-like domain of HB-EGF and that this interaction is specific for human HB-EGF.

Diphtheria toxin (DT) receptor associates with a 27-kD membrane protein (DRAP27) in monkey Vero cells. A cDNA encoding DRAP27 was isolated, and its nucleotide sequence was determined. The deduced amino acid sequence revealed that DRAP27 is the monkey homologue of human CD9 antigen. DRAP27 is recognized by CD9 antibodies. A human-mouse hybrid cell line (3279-10) possessing human chromosome 5, sensitive to DT, but not expressing CD9 antigen, was used for transfection experiments with DRAP27. When the cloned cDNA encoding DRAP27 was transiently expressed in 3279-10 cells, the total DT binding capacity was three to four times higher than that of untransfected controls. Transfectants stably expressing DRAP27 have an increased number of DT binding sites on the cell surface. Furthermore, the transfectants are 3-25 times more sensitive to DT than untransfected cells, and the sensitivity of these cells to DT is correlated with the number of DRAP27 molecules on the surface. However, when the cloned cDNA was introduced into mouse cell lines that do not express DT receptors, neither an increased DT binding nor enhancement of DT sensitivity was observed. Hence, we conclude that DRAP27 itself does not bind DT, but serves to increase DT binding and consequently enhances DT sensitivity of cells that have DT receptors. 12 proteins related to DRAP27/CD9 antigen were found through homology search analysis. These proteins appear to belong to a new family of transmembrane proteins.

Remarkable enhancements of the critical current density Jcwere obtained in polycrystalline La1.85Sr0.15CuO4with the splayed configuration of the columnar defects which were introduced by 3.5 GeV Xe ion irradiation with splay angles θ = 0°, ±5° and ±10°. In addition Jcincreased with the irradiation dose from 1.0×1011to 3.0×1011ions/cm2. Larger upward shift of the irreversibility line for the splayed configuration, in comparison with conventional parallel configuration, was observed. The results suggest that the vortex motion in La1.85Sr0.15CuO4is suppressed effectively by the splayed irradiation.

The behavior of vortex system in the presence of columnar defects in La1.85Sr0.15CuO4single crystal is studied by AC susceptibility measurements. It is found that the imaginary part χ″of the susceptibility as a function of temperature has a single peak, consistent with the Bose-glass theory, forBBcr, χ″exhibits double peaks, indicating the liquid-glass transition proceeds via two steps. The origin of the successive transitions is discussed.

Three reaction pathways yielding the triphenodithiazines (V) are shown in the reaction scheme.

EVIDENCE of increased adrenocortical secretory activity elicited by muscular exercise has been provided by indirect evaluations such as fall in ascorbic acid or cholesterol content of the adrenal gland1–3. However, no direct measurements of corticosteroid secretion in response to muscular exercise have been reported. The present studies were undertaken to evaluate quantitatively the effect of muscular exercise on adrenocortical activity by estimating directly the rate of secretion of adrenal 17-hydroxycorticosteroid in unanæsthetized dogs.