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Six coals with different ranks and different ash contents have been used to study the effect of demineralization on N 2 formation during coal pyrolysis. Chars obtained after pyrolysis have been also gasified with carbon dioxide at 1000 °C to investigate the influence of the demineralization on char gasification reactivity. The pyrolysis results show that the demineralization by acid washing drastically changes N 2 formation profiles and decreases nitrogen conversion to N 2 for low rank coals; on the other hand, the demineralization has little effect on N 2 formation for high rank coals. Addition of 0.5 wt% Fe promotes N 2 formation from the demineralized coals, but the catalytic effect depends on the coal type. It is found that the Fe remarkably promotes N 2 formation from the demineralized low rank coals, but the effect is much smaller for high rank demineralized coals. These observations suggest that the existing state of Fe-containing minerals and added Fe catalyst is important for catalytic N 2 formation during coal pyrolysis. Gasification results show that the demineralization lowers char gasification reactivity not only for low rank coals but also for high rank coals.

Summary A bovine/hamster hybrid cell panel consisting of 30 independent hybrids was developed to locate genes. Polymerase chain reaction analysis of 279 microsatellites on the cattle linkage map in this panel revealed the presence of all chromosomes in either entire or fragmented form. Among primer pairs prepared from bovine 3′-expressed sequence tags (ESTs), 1400 ESTs were assigned to specific chromosomes, of which 1303 were newly assigned in this study, and mapped 854 (61%) to 1 of 192 chromosomal segments using this panel. The regional mapping of new genes to cattle chromosomes can be rapidly achieved using this panel.

Breast cancer resistance protein (BCRP), an ATP-binding cassette transporter, confers resistance to a series of anticancer reagents such as mitoxantrone, 7-ethyl-10-hydroxycamptothecin, and topotecan. We reported previously that estrone and 17beta-estradiol reverse BCRP-mediated multidrug resistance. In the present study, we demonstrate that BCRP exports estrogen metabolites. First, we generated BCRP-transduced LLC-PK1 (LLC/BCRP) cells, in which exogenous BCRP is expressed in the apical membrane, and investigated transcellular transport of 3H-labeled compounds using cells plated on microporous filter membranes. The basal-to-apical transport (excretion) of mitoxantrone, estrone, and 17beta-estradiol was greater in LLC/BCRP cells than in LLC-PK1 cells. Thin-layer chromatography of transported steroids revealed that the transport of estrone and 17beta-estradiol was independent of BCRP expression. Alternatively, increased excretion of estrone sulfate and 17beta-estradiol sulfate was observed in LLC/BCRP cells. BCRP inhibitors completely inhibited the increased excretion of sulfated estrogens across the apical membrane. Conversion of estrogens into their sulfate conjugates was similar between LLC/BCRP and LLC-PK1 cells, suggesting that the increased excretion of estrogen sulfates was attributable to BCRP-mediated transport. Next, the uptake of 3H-labeled compounds in membrane vesicles from BCRP-transduced K562 (K562/BCRP) cells was investigated. 3H-labeled estrone sulfate, but not 3H-labeled estrone or 17beta-estradiol, was taken up by membrane vesicles from K562/BCRP cells, and this was ATP-dependent. Additionally, BCRP inhibitors suppressed the transport of estrone sulfate in membrane vesicles from K562/BCRP cells. These results suggest that BCRP does not transport either free estrone or 17beta-estradiol but exports sulfate conjugates of these estrogens.

Hydrodesulfurization (HDS) of dibenzothiophene (DBT) over laboratory synthesized bulk MoS2 catalysts using batch microautoclave reactor was studied. Influence of H2S (wide range level) on the DBT HDS activity and selectivity was investigated. In general, H2S was found to inhibit the direct desulfurization route that leads to biphenyl regardless the catalyst applied. However, the hydrogenation pathway which mainly produces phenylcyclohexane inferred to be either uninhibited or even more interestingly highly promoted by the presence of H2S depending on the catalyst nature. The ultimate sum impact of H2S on the catalytic activity shows two extremes, inhibition and promotion. Mechanism of H2S effect is proposed.

The mechanism for the cellular extrusion of sulfated conjugates is still unknown. In the present study, we investigated whether human wild type ABCG2 transports estrone 3-sulfate (E1S) using membrane vesicles from cDNA-transfected mouse lymphoma cell line (P388 cells). The uptake of [3H]E1S into ABCG2-expressing membrane vesicles was stimulated by ATP, and the Km value for [3H]E1S was determined to be 16.6 microm. The ABCG2-mediated transport of [3H]E1S was potently inhibited by SN-38 and many sulfate conjugates but not by glucuronide and glutathione conjugates or other anionic compounds. Other sulfate conjugates such as [3H]dehydroepiandrosterone sulfate (DHEAS) and [35S]4-methylumbelliferone sulfate (Km = 12.9 microm) and [35S]6-hydroxy-5,7-dimethyl-2-methylamino-4-(3-pyridylmethyl)benzothiazole (E3040) sulfate (Km = 26.9 microm) were also transported by ABCG2. Although [3H]methotrexate, [3H]17beta-estradiol-17beta-D-glucuronide, [3H]2,4-dinitrophenyl-S-glutathione, and [14C]4-methylumbelliferone glucuronide were transported by ABCG2, this took place to a much lesser extent compared with [3H]E1S. It was suggested that ABCG2 preferentially transports sulfate conjugates and that E1S and DHEAS are the potential physiological substrates for this transporter.

STI571, an Abl-specific tyrosine kinase inhibitor, selectively kills Bcr-Abl-containing cells in vitro and in vivo. However, some chronic myelogenous leukemia (CML) cell lines are resistant to STI571. We evaluated whether STI571 interacts with P-glycoprotein (P-gp) and multidrug resistance protein 1 (MRP1), and examined the effect of agents that reverse multidrug resistance (MDR) on the resistance to SI571 in MDR cells. STI571 inhibited the [(125)I]azidoagosterol A-photolabeling of P-gp, but not that of MRP1. K562/MDR cells that overexpress P-gp were 3.67 times more resistant to STI571 than the parental Philadelphia-chromosome-positive (Ph +) CML K562 cells, and this resistance was most effectively reversed by cepharanthine among the tested reversing agents. The concentration of STI571 required to completely inhibit tyrosine phosphorylation in K562/MDR cells was about 3 times higher than that in K562 cells, and cepharanthine abolished the difference. In KB-G2 cells that overexpress P-gp, but not Bcr-Abl, 2.5 micro M STI571 partly reversed the resistance to vincristine (VCR), paclitaxel, etoposide (VP-16) and actinomycin D (ACD) but not to Adriamycin (ADM) or colchicine. STI571 increased the accumulation of VCR, but not that of ADM in KB-G2 cells. STI571 did not reverse resistance to any agent in KB/MRP cells that overexpress MRP1. These findings suggest that STI571 is a substrate for P-gp, but is less efficiently transported by P-gp than VCR, and STI571 is not a substrate for MRP1. Among the tested reversing agents that interact with P-gp, cepharanthine was the most effective agent for the reversal of the resistance to STI571 in K562/MDR cells. Furthermore, STI571 itself was a potent reversing agent for MDR in P-gp-expressing KB-G2 cells.

Two model coals, one containing pyrrolic nitrogen and another containing pyridinic nitrogen, have been used to study N2 formation from different nitrogen structures during coal pyrolysis. The formation of N2 during pyrolysis of the model coals with and without a catalyst has been investigated in a fixed-bed reactor at 2.5−10 °C/min up to 1000 °C. The pyrolysis results show that calcium and iron catalyze both pyrrolic and pyridinic nitrogen in model coals to N2. Calcium-catalyzed N2 formation occurs at temperatures higher than 800 °C; however, the iron catalyzes N2 formation at temperatures below 800 °C. The remarkable formation of N2 results in low residual nitrogen in char. It is suggested that the calcium and iron catalysts catalyze N2 formation mainly through solid-phase reactions of pyrrolic and pyridinic nitrogen forms in char.

Breast cancer resistance protein (BCRP/ABCG2) is a novel member of ATP- binding cassette transporters, which induce multidrug resistance in cancer cells. We found that a high level of BCRP expression in CD4+ T cells conferred cellular resistance to human immunodeficiency virus type-1 (HIV-1) nucleoside reverse transcriptase inhibitors. The cell line MT-4/DOX 500 was established through the long-term culture of MT-4 cells in the presence of doxorubicin (DOX) and had reduced sensitivity to not only DOX but also zidovudine (AZT). MT-4/DOX 500 cells showed reduced intracellular accumulation and retention of DOX and increased ATP-dependent rhodamine 123 efflux. The cells were also resistant to several anticancer agents such as mitoxantrone, 7-ethyl-10-[4-(1-piperidino)-1-piperidino]carbonyloxycamptothecin, and 7-ethyl-10-hydroxycamptothecin. AZT was 7.5-fold less inhibitory to HIV-1 replication in MT-4/DOX 500 cells than in MT-4 cells. Furthermore, the anti-HIV-1 activity of lamivudine was severely impaired in MT-4/DOX 500 cells. In contrast, the antiviral activity of non-nucleoside reverse transcriptase inhibitors and protease inhibitors was not affected in the cells. MT-4/DOX 500 cells expressed glycosylated BCRP but not P-glycoprotein (ABCB1), multidrug resistance protein 1, 2, or 4 (ABCC1, -2, or -4), or lung resistance-related protein. In addition, the BCRP-specific inhibitor fumitremorgin C completely abolished the resistance of MT-4/DOX 500 cells to AZT as well as to DOX. An analysis for intracellular metabolism of AZT suggests that the resistance is attributed to the increase of ATP-dependent efflux of its metabolites, presumably AZT 5'-monophosphate, in MT-4/DOX 500 cells.

Bulk molybdenum sulfide catalyst was prepared by thermal decomposition of ammonium heptamolybdate at 800 °C in a flow of a 10% (v/v) H2S/H2 gas mixture. Investigation of the catalytic activity of the grinding catalyst for hydrodesulfurization (HDS) of dibenzothiophene (DBT) was carried out in a batch magnetically stirring microautoclave reactor at 340 °C and 3 MPa H2. The influence of a wide range of H2S concentration on the catalytic activity was studied. A significant promoting effect on HDS of DBT by H2S was observed. The experimental results showed the great enhanced contribution from the hydrogenation route with H2S in HDS of DBT. However, severe inhibition by H2S against the direct desulfurization route was observed. Kinetic analysis leads to the individual reaction rate constant in the reaction scheme performed.

Changes in the nitrogen functionality of 15 N-enriched condensation products prepared from glucose and 15 N-glycine were investigated during pyrolysis at 600-1000 °C. The structural changes in the condensation products were studied by means of solid-state 13 C and 15 N NMR spectroscopies. During pyrolysis, the aliphatic moieties of the condensation products decomposed and evolved as gas and tar. At pyrolysis temperatures above 600 °C, almost all the carbon in the chars were converted to aromatic carbon. After pyrolysis, large amounts of nitrogen remained in the chars as char nitrogen (char-N), and about 30% of the nitrogen was eliminated from the chars as HCN and NH 3 . With increasing temperature, the production of HCN and NH 3 increased and the amount of char-N decreased. By combining X-ray photoelectron spectroscopy and NMR results, detailed results for nitrogen fractions in chars were obtained. During pyrolysis, the fraction of unsubstituted pyrrole-N decreased and the fraction of quaternary-N increased. The fraction of pyridine-N remained almost constant at temperatures below 800 °C, but at 900 °C and above, the fraction of pyridine-N decreased. The fraction of substituted pyrrole-N showed minimum at 800 °C. On the basis of these results, structural changes of nitrogen functional groups during pyrolysis are discussed.

Chromosome-specific libraries provide a means to isolate genetic markers from specific chromosomal regions. A small-insert BTA6 library, constructed by microdissection, was screened for dinucleotide repeats (CA)15 and (GA)15. A total of 47 new microsatellite loci were developed and tested for polymorphism and informativeness using the MARC bovine mapping family.

A newly synthesized 1,4-benzothiazipine derivate, 4-[3-(4-benzylpiperidin-1-yl) propionyl]-7-methoxy-2,3,4,5-tetrahydro-1, 4-benzothiazepine monohydrochloride (JTV-519) was examined for its ability to reverse P-glycoprotein (P-gp) and multidrug resistance protein 1 (MRP1) mediated multidrug resistance (MDR) in K562/MDR and KB/MRP cells, respectively. JTV-519 at 3 microM reversed the resistance of K562/MDR cells to vincristine (VCR), taxol, etoposide (VP16), adriamycin (ADM) and actinomycin D and at 0.5 or 1 microM reversed their resistance to STI571. JTV-519 at 10 microM enhanced the accumulation of ADM in K562/MDR cells to the level in parental K562 cells and inhibited the efflux of ADM from K562/MDR cells. Photoaffinity labeling of P-gp with 3H-azidopine was almost completely inhibited by 500 microM JTV-519. JTV-519 at 3 microM also partially reversed the resistance of KB/MRP cells to VCR and at 500 microM partially inhibited the photoaffinity labeling of MRP1 with (125)I-II-azidophenyl agosterol A (125I-azidoAG-A). These results suggest that JTV-519 reversed the resistance to the anti-cancer agents in P-gp and MRP1 overexpressing multidrug-resistant cells by directly binding to P-gp and MRP1, and competitively inhibiting transport of the anti-cancer agents.

DNA topoisomerase I (Topo1) manages the topological state of DNA. Cleavable complexes, the covalent Topo1-DNA intermediates, become DNA damaged when the catalytic cycles are inhibited by the anti-tumor drug camptothecin (CPT). Intriguingly, Topo1 is modified rapidly and extensively with SUMO-1, a ubiquitin-like protein, in response to CPT. This study shows that the sumoylation enhances the cleavable complex formation and apoptosis induced by CPT. Indeed, substitutions of Lys117 and Lys153, identified as Topo1 sumoylation sites, reduced the CPT-induced cleavable complexes without influencing its in vitro catalytic activity. Consistent with this observation, CPT-induced cleavable complexes of wild-type Topo1 increased in a sumoylation-dependent manner. We also found that Topo1 sumoylation occurred independently of CPT when Topo1 was inactivated by mutation of the catalytic Tyr723. These findings suggested that Topo1 inactivation by CPT treatment can trigger Topo1 sumoylation, leading to enhanced cleavable complex formation.

Chondrodysplastic dwarfism in Japanese brown cattle is an autosomal recessive disorder characterized by short limbs. Previously, we mapped the locus responsible for the disease on the distal end of bovine chromosome 6. Here, we narrowed the critical region to ≈2 cM by using linkage analysis, constructed a BAC and YAC contig covering this region, and identified a gene, LIMBIN ( LBN ), that possessed disease-specific mutations in the affected calves. One mutation was a single nucleotide substitution leading to an activation of a cryptic splicing donor site and the other was a one-base deletion resulting in a frameshift mutation. Strong expression of the Lbn gene was observed in limb buds of developing mouse embryos and in proliferating chondrocytes and bone-forming osteoblasts in long bones. These findings indicate that LBN is responsible for bovine chondrodysplastic dwarfism and has a critical role in a skeletal development.

Breast cancer resistance protein (BCRP), an adenosine triphosphate-binding cassette transporter, confers resistance to a series of anticancer reagents, including mitoxantrone, SN-38 and topotecan. In the present study, we found that estrone and 17beta-estradiol potentiated the cytotoxicity of mitoxantrone, SN-38 and topotecan in BCRP-transduced K562 cells (K562 / BCRP). These estrogens showed only a marginal effect, or none, in parental K562 cells. Estrone and 17beta-estradiol increased the cellular accumulation of topotecan in K562 / BCRP cells, but not in K562 cells, suggesting that these estrogens inhibit the BCRP-mediated drug efflux and overcome drug resistance.

Formation of N2 during slow heating rate, temperature-programmed pyrolysis of a brown coal and a biodegraded leaf has been studied with a fixed-bed reactor in high-purity He. Formation profile of N2 depends strongly on type of sample. Demineralization by acid washing drastically changes the formation profiles and decreases nitrogen conversion to N2 for both the samples. Addition of iron to the demineralized brown coal promotes N2 formation and restores the profile to that of its original one. On the other hand, addition of calcium catalyst onto the demineralized biodegraded leaf promotes N2 formation and restores the profile to that of its raw sample. These results lead to the conclusion that inherent iron in the brown coal or inherent calcium in the biodegraded leaf is responsible for its remarkable formation of N2, respectively.

直留軽油 (LGO, S: 12,000ppm), 深度脱硫軽油 (DGO, S: 147ppm), 超深度脱硫軽油 (HDGO, S: 18.5ppm) および接触分解軽油 (LCO, S: 2600ppm) に含まれるジベンゾチオフェン (DBT) 類を定性•定量分析した。LGO中のDBT類の濃度は4000ppmで, 化学発光硫黄検出器によるガスクロマトグラムには約100本のDBT類のピークが認められた。DGO中の主要DBT類として4,6-ジメチルDBTおよび4-エチル-6-メチルDBTのほかに, 1,4,6-, 2,4,6-および3,4,6-トリメチルDBT類, 4,6-ジエチルDBT, 1-エチル-, 2-エチル-および3-エチル-4,6-ジメチルDBT類, および4位と6位にメチル基を有するテトラメチルDBT類が認められた。HDGO中の主要DBT類およびそれらの相対濃度はDGOにほぼ等しかった。LCO中のDBT類の濃度は2000ppmで, その組成はLGOに類似していた。

Coal -derived liquid obtained from 150ton/day NEDOL pilot plant was upgraded over a presulfided Ni-W/Al2O3 catalyst under a hydrogen pressure of 12MPa. The original coal liquid and three upgraded oils were separated into several compound groups and analyzed by GC -FID/NPD, GCMS and FIMS in order to identify the molecular species in the feedstock and upgraded oils.The coal liquid was composed of paraffins (17%), naphthenes (20%), oleffins (4%), monoaromatics (17%), polyaromatics (17%) and polar components (25%). After the upgrading over Ni-W/Al2O3, polyaromatics decreased to 0.7-4.7% and polar components were reduced below 0.1%, which resulted in the large increase of naphthenes and naphthenobenzenes. Basic nitrogen compounds (N: 4, 200ppm) in the coal liquid were piridines, anilines, quinolines and benzoquinolines and their hydrogenated derivatives, and nonbasic nitrogen compounds (N: 1, 400ppm) were alkylnitriles, pyrroles, indoles and carbazoles. Sterically hindered nitrogen compounds like 2, 4, 6-trimethylpyridines and 2, 6-anilines tended to remain in upgraded oils. Saturated cyclic nitrogen compounds of which molecular weights were expressed as 165+14n and 205+14n (n=0, 1, 2 …) were found to be least reactive for the denitrogenation, although their detail structure was not identified. Polyaromatic compounds which remained in upgraded oils were biphenyls and dibenzofurans due to their less reactivity and dihydrophenalene and pyrene at their hydrogenation equiribrium.

Using target and reference fattened steer populations, the performance of genotype imputation using lower-density marker panels in Japanese Black cattle was evaluated. Population imputation was performed using BEAGLE software. Genotype information for approximately 40,000 single nucleotide polymorphism (SNP) markers by Illumina BovineSNP50 BeadChip was available, and imputation accuracy was assessed based on the average concordance rates of the genotypes, varying equally spaced SNP densities, and the number of individuals in the reference population. Two additional statistics were also calculated as indicators of imputation performance. The concordance rates tended to be lower for SNPs with greater minor allele frequencies, or those located near the ends of the chromosomes. Longer autosomes yielded greater imputation accuracies than shorter ones. When SNPs were selected based on linkage disequilibrium information, relative imputation accuracy was slightly improved. When 3000 and 10,000 equally spaced SNPs were used, the imputation accuracies were greater than 90% and approximately 97%, respectively. These results indicate that combining genotyping using a lower-density SNP chip with genotype imputation based on a population of individuals genotyped using a higher-density SNP chip is a cost-effective and valid approach for genomic prediction.

Combinatorial phage peptide libraries have been used to identify the ligands for specific target molecules. These libraries are also useful for identification of the specific substrates of various proteases. A substrate phage library has a random peptide sequence at the N-terminus of the phage coat protein and an additional tag sequence that enables attachment of the phage to an immobile phase. When these libraries are incubated with a specific enzyme, such as a protease, the uncleaved phage is excluded from the solution with tag-binding macromolecules. This provides a novel approach to define substrate specificity. The aim of this review is to summarize recent progress on the application of the substrate phage technique to identify specific substrates of proteolytic enzymes. As an example, some of our own experimental data on the selection and characterization of substrate sequences for thrombin, a serine protease, and membrane type-1 matrix metalloproteinase (MT1-MMP) will be presented. Using this approach, the canonical consensus substrate sequence for thrombin was deduced from the selected clones. As expected from the collagenolytic activity of MT1-MMP, a collagen-like sequence was identified in the case of MT1-MMP. A more selective substrate sequence for MT1-MMP was identified during a substrate phage screen. The delineation of the substrate specificity of proteases will help to elucidate the enzymatic properties and the physiological roles of these enzymes. Comprehensive screening of very large numbers of potential substrate sequences is possible with substrate phage libraries. Thus, this approach allows novel substrate sequences and previously unknown target molecules to be defined.

In order to study nitrogen release from different nitrogen structures during coal pyrolysis, two model coals containing pyrrolic or pyridinic nitrogen have been prepared using cellulose and phenolic nitrogen compounds. Nitrogen release during pyrolysis of the model coals without and with catalyst has been studied in a fixed-bed reactor at 50°C/min up to 1000°C. The pyrolysis results show that HCN rather than NH3 is formed from the decomposition of pyrrolic and pyridinic nitrogen, and iron catalyst is effective on nitrogen removal for pyrrolic and pyridinic nitrogen. It is especially noteworthy that calcium catalyst also promotes nitrogen removal significantly from both pyrrolic and pyridinic nitrogen.

In order to study nitrogen release from different nitrogen functionality, two types of model coals with different nitrogen structures have been prepared using cellulose and phenolic nitrogen compounds, 2-hydroxycarbazole and 6-hydroxyquinoline. Carbon and nitrogen structures in the model coals have been analyzed by solid-state 13CNMR, pyrolysis GC-NPD and GC-MS. The 13C-NMR spectra of the model coals very closely resemble those of natural coals with similar carbon content, and the pyrolysis GCNPD analyses show that the model coals have the same nitrogen-containing skeleton as the original nitrogen compounds.Addition of Ca and Fe catalysts on the model coals has also been carried out to make clear the effects of the Ca and Fe on nitrogen release from the model coals. Nitrogen release during pyrolysis of the model coals without and with catalyst has been studied in a fixed-bed reactor at 50°C/min up to 1, 000°C. The pyrolysis results show that thermal decomposition behavior of pyrrolic and pyridinic nitrogen in the model coals is similar, and both calcium and iron catalysts are effective on nitrogen removal from the model coals prepared at 200°C. However, the efficiency of nitrogen removal by the catalysts depends on the preparation temperature of the model coals.

ジベンゾチオフェン(DBT), 4-メチルDBT, 4,6-ジメチルDBTおよび2,8-ジメチルDBTのメチル化, エチル化, プロピル化およびイソプロピル化により下記のアルキル置換DBT類を調製し, 各化合物についてカラム保持指標と質量スペクトルの特徴 (最大フラグメントイオンに対する親イオンの割合) を求めた。調製したDBT類は110種で, その主なものはメチルDBT類, ジメチルDBT類, トリメチルDBT類, テトラメチルD-BT類, ペンタメチルDBT類, ヘキサメチルDBT類, エチルDBT類, ジエチルDBT類, トリエチルDBT類, エチル-メチルDBT類, ジエチル-メチルDBT類およびエチル-ジメチルDBT類であった。

It has been speculated that Indonesian cattle originated from indigenous Bali cattle and that its genetic structure was modified by the successive migration of Indian cattle. In order to study the genetic structure in Indonesian native cattle, the blood protein polymorphism in Bali cattle, Java cattle, Madura cattle, and Filial Ongole cattle were studied. Ten of the 25 loci examined, serum albumin (Alb), serum transferrin (Tf), serum post transferrin (P-tf), serum ceruloplasmin (Cp), serum amylase-I (Am-I), cell carbonic anhydrase (CA), hemoglobin α (Hb-α), hemoglobin β (Hb-β), cell NADH-diaphorase II (Dia-II), and cell peptidase-B (Pep-B) were found to be polymorphic.The proportion of polymorphic loci and average heterozygosity among the Bali cattle were lower than those among the other Indonesian cattle populations (Madura, Java and Filial Ongole) . The phylogenetic trees and principal component analysis revealed that the Bali cattle were clustered separately from the other three populations. However, specific alleles that were identified in Bali cattle were also found among the Madura, Java and Filial Ongole cattle. We suggest that Bali cattle had introgressed to other indigenous cattle widespread in Indonesia.

Defective xanthine dehydrogenase (XDH) activity in humans results in xanthinuria and xanthine calculus accumulation in kidneys. Bovine xanthinuria was demonstrated in a local herd and characterized as xanthinuria type II, similar to the Drosophila ma-l mutations, which lose activities of molybdoenzymes, XDH, and aldehyde oxidase, although sulfite oxidase activity is preserved. Linkage analysis located the disease locus at the centromeric region of bovine chromosome 24, where a ma-l homologous, putative molybdopterin cofactor sulfurase gene (MCSU) has been physically mapped. We found that a deletion mutation at tyrosine 257 in MCSU is tightly associated with bovine xanthinuria type II.

In previous studies, we established two camptothecin (CPT)-resistant sublines, HT-29 / CPT and St-4 / CPT, from the human colon cancer cell line HT-29 and the human stomach cancer cell line St-4, respectively. Cellular contents of DNA topoisomerase I (topo I) in the resistant cells were eight-fold less than those in the corresponding parental lines. In this study, we have shown expression of two species of the TOP1 mRNA in HT-29 / CPT. The longer mRNA (4.0 kb) is the wild-type TOP1 mRNA, and the shorter mRNA (3.3 kb) proved to have a deletion of 672 bp (nucleotides 58 - 729 or 59 - 730) that caused the in-frame deletion of amino acids 20 - 243 of human topo I. The deleted region is identical to exons 3 - 9 of the TOP1 gene. The expression level of the 3.3-kb mRNA was similar to that of the wild-type mRNA in HT-29 / CPT. St-4 / CPT expressed only the wild-type TOP1 mRNA in lesser amounts than did St-4. Mouse NIH3T3 cells transfected with the wild-type TOP1 cDNA showed higher sensitivity to CPT than the parental cells, whereas those transfected with the deleted TOP1 cDNA showed levels similar to those of the parental cells. Expression of the exogenous TOP1 mRNA was confirmed; however, expression of the truncated topo I was not detected in cells transfected with the deleted TOP1 cDNA. These results suggest that the expression of the deleted TOP1 mRNA led to the low expression of CPT-sensitive topo I in the resistant cells.

Recycle solvents used in a 150t/d NEDOL coal liquefaction pilot plant were separated into non-polar and polar component, and the non-polar component were examined by GC and GC-MS. Major compound classes in the each non-polar component were quantified, and the influences of raw coal and the concentration of coal in a feed slurry on their composition were discussed.The influence of raw coal was appreciable in the amount of n-paraffins and polar component in the solvent. Perhydrogenated homologues of polycondensed aromatic compounds were contained in only negligible amounts. The higher concentration of coal in a feed slurry needed the larger amount of heavy distillate in the solvent and the severe hydrotreatment to enhance hydrogen donating ability of the solvent, which resulted in the increase of larger cyclic compounds with more naphthenic rings.The fa value of each compound class was similar for all the solvents, and the average fa values were useful for the evaluation of the fa values of the recycle solvents by being combined with the compound class composition obtained from HPLC

Nonhuman primate models are useful to evaluate the safety and efficacy of new therapeutic modalities, including gene therapy, before the inititation of clinical trials in humans. With the aim of establishing safe and effective approaches to therapeutic gene transfer, we have been focusing on a small New World monkey, the common marmoset, as a target preclinical model. This animal is relatively inexpensive and easy to breed in limited space. First, we characterized marmoset blood and bone marrow progenitor cells (BMPCs) and showed that human cytokines were effective to maintain and stimulate in culture. We then examined their susceptibility to transduction by retroviral vectors. In a mixed culture system containing both marmoset stromal cells and retroviral producer cells, the transduction efficiency into BMPCs and peripheral blood progenitor cells (PBPCs) was 12% to 24%. A series of marmosets then underwent transplantation with autologous PBPCs transduced with a retroviral vector carrying the multidrug resistance 1 gene (MDR1) and were followed for the persistence of these cells in vivo. Proviral DNA was detectable by polymerase chain reaction (PCR) in peripheral blood granulocytes and lymphocytes in the recipients of gene transduced progenitors up to 400 days posttransplantation. To examine the function of the MDR1 gene in vivo, recipient maromsets were challenged with docetaxel, an MDR effluxed drug, yet the overall level of gene transfer attained in vivo (

To determine the formation and reactivity of addition compounds produced during hydrodenitrogenation (HDN), we investigated the HDN of isoquinoline for a sulfided Ni–Mo/Al 2 O 3 catalyst operated under a hydrogen pressure of 12 MPa (cold charge) in the temperature range 300–375°C. The reaction products were classified into five groups of compounds: 1. hydrogenated derivatives of isoquinoline (tetrahydroisoquinolines, decahydroisoquinolines, and their isomers); 2. nitrogen-containing ring-opened products (1-amino-2-(2-methylphenyl)ethane and 1-amino-1-(2-ethylphenyl)methane); 3. denitrogenated products (1-ethyl-2-methylbenzene, 1-ethyl-2-methylcyclohexane, and their isomers); 4. addition products (hydrocarbons with molecular weights of 238, 244, and 250 and nitrogen-containing compounds with molecular weights of 249, 251, and 257); and 5. cracked products (toluene, ethylbenzene, dimethylbenzenes, and their hydrogenated derivatives). Most of the nitrogen-containing addition compounds appear to be substituted on the nitrogen atom. The HDN of isoquinoline was more than 10 times faster than the HDN of quinoline, whereas the hydrogenation of isoquinoline was difficult compared to the hydrogenation of quinoline. The reaction network for the HDN of isoquinoline is also presented.

In previous studies, we isolated a mutant DNA topoisomerase I cDNA from a camptothecin (CPT)-resistant human T-lymphoblastic leukemia cell line, CPT-K5, and demonstrated that an amino acid change from Asp to Gly at residue 533 is responsible for the CPT resistance of the enzyme. In the present study, we have constructed a bicistronic retroviral vector, Ha-TM1-IRES-neo, that carries the mutant (Gly-533) TOP1 cDNA (TM1) and a neomycin-resistance gene to examine the effect of mutant DNA topoisomerase I (topo I) expression on CPT resistance of cells. HeLa S3 cells were transduced with Ha-TM1-IRES-neo, and the transduced cells were selected with G418. Two independently isolated populations of the G418-resistant cells and 2 clones showed 1.7- to 1.8-fold higher resistance to CPT than the control cells. Integration and expression of the exogenous TOP1 were confirmed by genomic and RT-PCR analyses. The topo I enzyme (mixture of mutant and wild-type) expressed in the transduced cells showed 3-fold resistance to CPT in cleavable-complex-formation assay and DNA-relaxation assay. Mutant topo I activity in the transduced cells was as much as 10% that of the endogenous enzyme. Our results clearly show that expression of Gly-533 topo I confers a dominant form of CPT resistance in cells expressing wild-type topo I. The mutant TOP1 could be used for the protection of normal bone marrow cells of cancer patients from the severe hematotoxicity of CPT-derivative anti-tumor agents.

Recent plant materials, peat and coals were compressed and thermally treated at various temperatures (200-400°C) for 25 hours. The compaction pyrolysis products were observed with a microscope in both reflected and fluorescent lights, and their vitrinite reflectance (Ro) were also measured.The Ro values of the original samples varied from 0.1 of peat to 0.7 of Miike bituminous coal. After the thermal treatment at 300, 350 and 400°C, the Ro values increased to 0.7-1.0, 1.2-1.4 and 1.7-2.1, respectively, in spite of the kind of the original material. The compaction pyrolysis products obtained from plant materials at 200°C showed specific textures which are often observed in natural low rank coals. Those textures became homogeneous with the increasing temperatures of the thermal treatment. Petrographic characteristice of the xylems of Morwell coal and red pine changed similarly during the thermal treatment. Because peat and humus soil contained a large amount of mineral matter, they provided the pyrolysis products with different textures from other materials.The fluorescent material of leaf (metasequoia) after the thermal treatment at 200°C was considered as cutinite, and it started to melt around 250°C and disappeared around 300°C, whereas thet of Miike bituminous coal did not melt and disappear even around 300°C.There still remain quite a few coal macerals of which origin and formation are unclear, and the artificial maturation followed by the microscopic investigation is one of effective approaches to elucidate them.

Chronic granulomatous disease (CGD) is a group of disorders characterized by the failure of phagocytes to produce superoxide. One-third of the cases of CGD in the USA and Europe results from defects in the gene encoding p47phox, a cytoplasmic component of NADPH oxidase for superoxide generation. In this study, we constructed the bicistronic retrovirus vector Ha-MDR-IRES-p47, which carries cDNAs for a human multi-drug-resistance gene (MDR1) and p47phox. The amphotropic retroviral producer cells were generated, and the supernatant of the producer cells was used to transduce Epstein-Barr virus-transformed B (EBV-B) cells, established from B cells of p47phox-deficient CGD patients, as an in vitro model of gene therapy for p47phox-deficient CGD. The transduced cells expressed both P-glycoprotein and p47phox protein, and the expression levels were increased after appropriate vincristine selection. The levels of superoxide production in the vincristine-selected cells were increased to a level similar to normal EBV-B cells. This result suggests that it is possible to achieve 100% correction of the CGD defect in p47phox-deficient EBV-B cells by using the bicistronic vector. This strategy could be employed not only in vitro, but also in vivo, in the gene therapy of a number of inherited diseases.

Retrovirus-mediated transfer of O 6 -methylguanine-DNA methyltransferase (MGMT; E.C. 2.1.1.63) and a human multidrug-resistance gene ( MDR1 ) confers resistance to nitrosoureas and natural product antitumor agents, respectively. In a previous study, we constructed two bicistronic retroviral vectors, Ha-MDR-IRES-MGMT and Ha-MGMT-IRES-MDR, that allow co-expression of the MGMT gene and the MDR1 gene to protect cells from the toxicity of combination chemotherapy. Each cell transduced with Ha-MDR-IRES-MGMT or Ha-MGMT-IRES-MDR showed high-level resistance to vincristine and 1-(4-amino-2-methyl-5-pyrimidinyl)methyl-3-(2-chloroethyl)-3-nitrosourea (ACNU), indicating that the two drug-resistance genes can be functionally co-expressed from these vectors. In the present study, we examined whether the expression of MGMT from these MDR1 – MGMT bicistronic retroviral vectors could protect cells from the genotoxicity of nitrosoureas. Three independent Ha-MDR-IRES-MGMT-transduced clones and three independent Ha-MGMT-IRES-MDR-transduced clones of HeLa MR cells showed 12-23-fold and 27–30-fold higher MGMT activity than the parental cells. These clones are more resistant to ACNU mutagenicity measured by the frequency of the emergence of 6-thioguanine-resistant colonies after ACNU treatment over the frequency seen in the parental cells. The ACNU-induced sister chromatid exchange (SCE) was markedly suppressed in these clones. Murine bone marrow cells were transduced with either Ha-MDR-IRES-MGMT or Ha-MGMT-IRES-MDR. Non-selected populations of the transduced cells showed only marginal increases in drug resistance and MGMT activity. Remarkable increase in drug resistance and MGMT activity were observed after a short exposure of the transduced cells to vincristine. The Ha-MDR-IRES-MGMT-transduced, vincristine-selected bone marrow cells showed 27-fold resistance to vincristine, 7-fold resistance to ACNU, and 10-fold higher MGMT activity than the non-transduced, non-selected cells. The Ha-MGMT-IRES-MDR-transduced, vincristine-selected cells showed 8-fold resistance to vincristine, 16-fold resistance to ACNU and 19-fold higher MGMT activity than the non-transduced, non-selected cells. The rates of ACNU-induced SCE in the vincristine-selected cells were as follows: non-transduced cells (non-selected) and HaMDR-transduced cells>Ha-MDR-IRES-MGMT-transduced cells>Ha-MGMT-IRES-MDR-transduced cells. Again, the only marginal levels of increases in the rates of ACNU-induced SCE were observed in non-selected population of the transduced cells. These results indicate that the MDR1 – MGMT bicistronic retrovirus vectors would be useful to protect normal hematopoietic cells from nitrosourea-induced mutagenesis, and drug-selectable bicistronic constructs would have great advantage over non-selectable vectors.

The reacivites of preasphaltenes obtained by liquefaction of Illinois No.6 coal, PA (I), and Morwell brown coal, PA (M), were studied for hydrotreating in the presence or absence of a catalyst under hydrogen pressure of 8 MPa (coal charge) and temperature range from 320 to 440°C. The feed preasphaltenes were easily converted to THE insoluble fraction during preservation at room temperature; 35 wt% of THFI from PA (I) and 60wt% THFI from PA (M). In the reaction of PA (I) without catalyst, craking of THEI to PA, cracking of PA to asphaltene and polymerization of PA to THFI were the major reactions and the production of hexane soluble fraction (HS) was scarcely observed. Especially, polymerization of PA to THEI was very remarkable at 440°C. The presence of catalyst promoted reactions to produce asphaltene and HS and prevented the formation of THFI. In the reaction of PA (M), rather large amout of HS was produced when catalyst was not used. The productions of asphaltene and HS increased by the existence of catalyst, whereas, similar yields of THEI were obtained in both cases of presence and absence of catalyst at 440°C, which suggests that catalyst may be deactivated easfly by the PA (M) reaction at 440°C.

100 samples of recycle solvent were obtained from continuous opera-tion (50 days) of 1 ton/day coal liquefaction plant at Kimitsu. Phenanthrene and anthracene derivatives in the recycle solvents were analyzed in detail by the technique of selective ion measurement GC/MS. According to the replacement of the initial solvent by coal-derived products, the amounts of phenanthrene derivatives decreased remark-ably, while those of anthracene derivatives decreased slightly. For phenanthrene deriv-atives, octahydrophenanthrenes play an important role as hydrogen donors by releasing eight hydrogen atoms in the recycle solvent, on the other hand for anthracene deriva-tives, tetrahydroanthracene plays an important role by releasing four hydrogen atoms. This results indicate that hydrogen donatable ability of phenanthrene derivatives is twice as large as the ability of anthracene derivatives.

100 samples of recycle solvent were obtained by continuous opera-tion (50 days) of 1 ton/day coal liquefaction plant at Kimitu. Acenaphthene and biphenyl derivatives in the recycle solvents were analyzed in detail by the technique of selective ion measurement of GC/MS. Because of the replacement of the initial solvent by coal-derived product during the continuous operation, the amounts of acenaphthenes (including hydrogenated derivatives) and biphenyls (including hydrogenated derivatives) in the recycle solvents decreased remarkably while the amounts of compounds substituted by methyl or ethyl groups increased. The difference of degree of hydrogenation before and after hydrotreating process of solvent was about 40%, 20% and 20% for acenaphthene, methylacenaphthenes and biphenyl, respectively, and any significant difference was not observed for methylbiphenyls. The hydrogenated derivatives of alkyl-substituted acenaphthenes and biphenyls with over two carbon atoms in the alkyl chains were not found in the solvent.

2-プロピルアニリン (2-PA), N-プロピルアニリン (N-PA) およびγ-フェニルプロピルアミン (γ-PPA) の反応をバッチ式小型オートクレーブを用い, 硫化Ni-Mo/Al2O3触媒上, 温度300～350°C, 水素初圧12MPa, 反応時間15～120minで行い, 含窒素化合物の構造と反応性の関係を調べた。生成物の組成は, (1) 分解を伴わない水素化物, (2) 脱窒素生成物, (3) 脱窒素中間体が共存する含窒素化合物に付加した付加生成物, および (4) 脱窒素生成物の再分解生成物の4化合物タイプに大別された。(1) として, 2-PAから2-プロピルシクロヘキサンアミンが, N-PAからN-プロピルシクロヘキサンアミンが生成し, γ-PPAの水素化物は認められなかった。(2) として, 2-PAからプロピルシクロヘキサンが, N-PAからプロパンとシクロヘキサンが, γ-PPAからプロピルベンゼンが主として生成した。(3) として, 2-PAから分子量259の化合物が, N-PAから分子量175, 177および181の化合物が, γ-PPAから分子量253の化合物が主として生成した。付加化合物の生成量はγ-PPAの反応で非常に高く, その再分解はγ-PPAの脱窒素に比べて困難であった。(4) はいずれの反応においてもわずかであった。反応全体に対するネットワークを提案した。

In the present study, we examined the effect of interleukin-2 (IL-2) gene transfer into multidrug resistance (MDR) cancer cells on the therapeutic efficacy of MRK16. Human MDR ovarian cancer cells, AD10, were transduced with a bicistronic IL-2 retrovirus, Ha-IL2-IRES-Neo. The G418-resistant population, IL2-AD10, secreted IL-2 into the culture supernatant and did not form a tumor mass in nude mice. The IL2-AD10 cells were more susceptible to the cytotoxicity of murine spleen cells than AD10 cells in vitro. For examination of the effect of IL-2 gene transfer on the antitumor activity of MRK16 against P-glycoprotein-positive tumors, IL2-AD10 cells were co-transplanted s.c. with AD10 cells into nude mice in a ratio of 1:3, and the mice were treated with MRK16 on days 2 and 7. MRK16 markedly inhibited the growth of AD10 cells mixed with IL2-AD10 cells under conditions (0.3-1 microgram/body) where it showed only marginal effects on the growth of AD10 tumors. These findings suggest that IL-2 gene transfer potentiates the antitumor activity of MRK16 against MDR tumors.

Recycle solvents were obtained in 50 d of continuous operation of a 1 ton/day coal-liquefaction plant at Kimitsu. Acidic fractions were obtained by base extraction for the starting solvent and final solvent, and their components were analyzed in detail by gas chromatography/mass spectrometry. The application of a selected ion monitoring technique has allowed the assignment of the atomic compositions to practically all compounds despite many of their presence as overlapping peaks on the chromatogram: 327 individual compounds for 142 peaks. These compounds have been classified into nine of compound types: phenols, indanols/tetrahydronaphthols, naphthols, biphenylols/acenaphthenols, hexahydroacenaphthylenols, fluorenols, phenanthrenols/anthracenols, phenylnaphthols, and pyrenols/fluoranthenols. As compared to the initial solvent, the final solvent contains higher concentrations of indanols/tetrahydronaphthols and hexahydroacenaphthylenols and contains a lower concentration of pyrenols/fluoranthenols.

石炭液化反応によつて得られた液化油中の粗ナフサ,それを精製処理した精製ナフサ,さらにそれをリホーミング処理した改質油および市販のレギュラーガソリソの組成を比較,検討した.石炭液化粗ナフサの組成は鎖状炭化水素類,単環炭化水素類,二環炭化水素類,含酸素化合物およびその他に大別された.粗ナフサは石油ガソリンに比べて,直鎖パラフィン類,シクロパラフィン類,シクロオレフィン類,ビシクロパラフィン類および含酸素化合物の濃度が高く,分枝パラフィン類,単環芳香族類の濃度が低かった.含酸素化合物の組成は,GCIAED-GC/MS分析の結果フェノール類の他にケトソ類が主成分であった.粗ナフサは水素化精製処理によりパラフィン類,シクロパラフィン類の濃度が増大し,これは主としてオレフィン類の水素化と含酸素化合物の水素化脱酸素反応によることが示唆された.改質油では直鎖パラフィン類,シクロパラフィン類および残存するフェノールの濃度が減少し,分枝パラフィン類,単環芳香族類の濃度が増大し,液化油の改質油はガソリンの主要混合基材として使用し得ることが示唆された.

Nucleolar protein p120 is considered to be associated with cell proliferation and has also been detected in a broad range of human malignant cells and tissues, but not in either normal resting tissue or most benign tumors. To clarify the significance of the expression of p120 in colorectal tumors or to evaluate the contribution of p120 in the development of colorectal carcinoma, the authors developed a monoclonal antibody against p120 and then examined its expression in adenoma, carcinoma, and normal mucosa. In adenomas, p120 expression was shown in none of 13 cases of mild dysplasia (0%), 2 of 15 of moderate dysplasia (13.3%), and in 2 cases of severe dysplasia (100%). p120-positive adenomas of moderate dysplasia tended to be larger and had higher Ki-67 indexes than the negative ones (adenomas of moderate dysplasia). All 27 carcinomas were positive for p120. p120 immunostaining was found in the nuclei and corresponded closely to the prominent nucleoli of tumor cells. In contrast, either weak or the occasional expression of p120 was traced in only one of the nine normal mucosae (11.1%). Three of the transitional mucosae of the carcinoma were also positive for p120. The percentage of p120-positive tumor cells (p120 index) ranged from 3.2% to 86.6%, and the mean p120 indexes of the four adenomas and all carcinomas were 21.3% and 41.5%, respectively. The p120 index was significantly related to the Ki-67 index (P < .001) in the p120-positive tumors, whereas the p120 index of the carcinoma did not significantly correlate to the known prognostic markers, such as tumor size, stage, or the degree of differentiation. These results thus suggest that the expression of p120 serves as a marker for cells with a high proliferative potential and is linked to the late events of colorectal tumor progression.

キノリンの水素化脱窒素を硫化ニッケル-モリブデン/Al2O3触媒を用い,温度300-375℃,水素圧力12MPa,反応時間15-120minで行い,反応ネットワークを検討した.325℃ 以下では主としてキノリンの水素化が起こり,脱窒素反応は350℃ 以上で顕著になった.反応生成物は,(1) キノリンの水素化物,(2) 脱窒素を伴わない開環生成物,(3) 脱窒素生成物の付加化合物,(4)脱窒素生成物および(5) 脱窒素生成物の再分解生成物の五つの型に大別された.(1) はテトラヒドロキノリン類,デカヒドロキノリン類およびそれらの異性体で,デカヒドロキノリン類の開環は比較的起こりにくかった.(2) は2-プロピルアニリン,1-アミノ-2-プロピルシクロヘキサンおよびそれらの異性体で,1-アミノ-2-プロピルシクロヘキサンは主として2-プロピルアニリンの水素化で生成した.(3) は分子量244の異性体4種類および分子量250の異性体5種類を主成分とする窒素を含まない化合物と,分子量253の異性体3種類,分子量257の異牲体7種類および分子量263の異性体10種類を主成分とする含窒素化合物であった.(3) の全収率は温度および時間と共に増大し,(3) の二次的分解は起こりにくかった.また窒素を含む(3) の脱窒素が進行するにつれ,それに対応して窒素を含まない(3) の濃度増大が認められ,脱窒素生成物は窒素化合物の炭素原子に付加することが示唆された(4) はプロピルシクロヘキサン,プロピルシクロヘキセン,プロピルベソゼンおよびそれらの異性体で,プロピルシクロヘキセンには6種類の異性体が認められた.(5) はシクロヘキサン,メチルシクロヘキサンおよびエチルシクロヘキサンであり,それらの合計収率は1%以下であった.以上の結果をまとめた反応ネットワークを提案した.

The recent decline in fertility is a serious problem in the dairy industry. To overcome this problem, we performed a genome-wide association study using 384 Holsteins and identified four loci associated with conception rates. Two of them contained gap junction-related genes: PKP2 and CTTNBP2NL. Further analysis confirmed that PKP2 increased connexin 43, a gap junction protein, whereas CTTNBP2NL dephosphorylated connexin 43. Knockdown of PKP2 or overexpression of CTTNBP2NL inhibited embryo implantation in mice. The other two loci contained neuroendocrine-related genes: SETD6 and CACNB2. Additional experiments indicated that SETD6 is involved in the transcriptional regulation of gonadotropin-releasing hormone, whereas CACNB2 controlled the secretion of follicle-stimulating hormone in cattle. The total allele substitution effect of these genes on conception rate was 3.5%. Our findings reveal important roles for gap junction communication and the neuroendocrine system in conception and suggest unique selection methods to improve reproductive performance in the livestock industry.

(-)-Sesamin is a lignan present in sesam oil and a number of medicinal plants. It exerts various pharmacological effects, such as prevention of hyperlipidemia, hypertension, and carcinogenesis. Moreover, (-)-sesamin has chemopreventive and anticancer activity in vitro and in vivo. Multidrug resistance (MDR) of tumors leads to fatal treatment outcome in many patients and novel drugs able to kill multidrug-resistant cells are urgently needed. P-glycoprotein (MDR1/ABCB1) is the best known ATP-binding cassette (ABC) drug transporter mediating MDR. ABCB5 is a close relative to ABCB1, which also mediates MDR. We found that the mRNA expressions of ABCB1 and ABCB5 were not related to the 50% inhibition concentrations (IC50) for (-)-sesamin in a panel of 55 cell lines of the National Cancer Institute, USA. Furthermore, (-)-sesamin inhibited ABCB1- or ABCB5-overexpressing cells with similar efficacy than their drug-sensitive parental counterparts. In addition to ABC transporter-mediated MDR, we attempted to identify other molecular determinants of (-)-sesamin resistance. For this reason, we performed COMPARE and hierarchical cluster analyses of the transcriptome-wide microarray-based mRNA expression of the NCI cell panel. Twenty-three genes were identified, whose mRNA expression correlated with the IC50 values for (-)-sesamin. These genes code for proteins of different biological functions, i.e. ribosomal proteins, components of the mitochondrial respiratory chain, proteins involved in RNA metabolism, protein biosynthesis, or glucose and fatty acid metabolism. Subjecting this set of genes to cluster analysis showed that the cell lines were assembled in the resulting dendrogram according to their responsiveness to (-)-sesamin. In conclusion, (-)-sesamin is not involved in MDR mediated by ABCB1 or ABCB5 and may be valuable to bypass chemoresistance of refractory tumors. The microarray expression profile, which predicted sensitivity or resistance of tumor cells to (-)-sesamin consisted of genes, which do not belong to the classical resistance mechanisms to established anticancer drugs.

Background The ATP-binding cassette (ABC) transporters P-glycoprotein (P-gp)/ABCB1 and breast cancer resistance protein (BCRP)/ABCG2 are involved in the intestinal absorption and renal excretion of various substrate drugs. Their activities affect sub-therapeutic drug concentrations and excretion of natural transporter substrates. The new oral anti-HCV drug telaprevir has dramatically improved the efficacy of hepatitis-C virus (HCV) treatment, and recent studies have suggested a possible pharmacological interaction between telaprevir and P-gp. We studied the kinetics of in vitro interactions between telaprevir and P-gp and BCRP to understand the molecular basis of that interaction. Findings The effect of telaprevir on P-gp- and BCRP-mediated transport was evaluated by an in vitro vesicle transporter assay using different transport substrates, and the kinetics of transporter inhibition was determined. The results showed that telaprevir could inhibit P-gp- and BCRP-mediated transport in the in vitro vesicle transport assay, with each IC50 values of ≈ 7 μmol/L and ≈ 30 μmol/L, respectively. Analyses of Lineweaver–Burk plots showed that telaprevir was likely to be a competitive inhibitor against P-gp and BCRP. Photoaffinity labeling experiments were employed to observe competitive inhibition by telaprevir using iodoarylazidoprazosin (IAAP) as a binding substrate for P-gp and BCRP. These experiments revealed that telaprevir inhibited [125I]-IAAP-binding with P-gp and BCRP. Conclusion Telaprevir competitively inhibited P-gp and BCRP, and P-gp-mediated transport was more sensitive to telaprevir compared with BCRP-mediated transport. These data suggest that telaprevir represses the transporter functions of P-gp and BCRP via direct inhibition.