Search

Effects of S-23142 on protoporphyrin IX (Proto IX) biosynthesis in chloroplasts isolated from Spinacia oleracea L. were examined using reverse phase HPLC with fluorescence monitoring. The synthesis of Proto IX was inhibited to a level of 50% by 10-9 ᴍ of S-23142 in this system. The effects of S-23142 was also tested in chloroplasts isolated from two types of photomixotrophic tobacco cells, wild type and S-23142 tolerant cells. The biosynthesis of both the wild type cells and YZI-1 S cells were inhibited at 50% by 10-9 ᴍ and 10-7 ᴍ of S-23142, respectively. It is, therefore evident that the mutation in the tolerant cell is associated with the Protox. To investigate the localization of Proto IX biosynthesis and the target site of S-23142, spinach chloroplasts were osmotically broken and separated into stroma and membrane (thylakoid and envelope) fractions. A very active Proto IX synthesis from ALA was found in the stromal fraction, while no activity of Proto IX synthesis was observed in the membrane fraction. These results suggest that most Proto IX synthetic activity and a target-site of S-23142 exist in the stromal fraction.

Heating the aqueous solutions of asparagine-containing dipeptides in which asparagine locates at their C-terminal gave sequential polypeptides having the molecular weight up to 3000–5000.

We examined changes in ability- and performance-based instrumental activities of daily living (IADL) among community-dwelling elderly a year after a baseline survey was taken.Community-dwelling elderly in Otama village were interviewed in 2006 and followed for one year. Based on ability- and performance-based IADL assessments regarding 3 daily tasks from the Kihon Checklist (using public transportation, shopping for daily necessities, and managing bank accounts), each was assigned to one of three performance groups: good, borderline, or low.One year after the survey, the proportion of those assigned to the good performance group decreased, while those assigned to either the borderline or low performance groups increased. Among those assigned to the good performance group, 75.6% remained in that group, 21.1% shifted to the borderline performance group, and 3.3% shifted to the low performance group one year later. Compared to those remaining in the good performance group, those who shifted to the borderline performance group had significantly lower muscular strength, less healthy attitudes, and lower IADL as measured by the Tokyo metropolitan Institute of Gerontology (TMIG) Index of Competence.Fewer elderly were assigned to the good performance group and more were assigned to the borderline and low performance groups one year after the survey. Shifts from the good to borderline performance group were common among those with lower physical status and less healthy attitudes. We conclude that ability- and performance-based evaluations are important tools to screen those in the borderline performance group requiring preventive care.

Newly synthesized glucose transporter 4 (GLUT4) enters into the insulin-responsive storage compartment in a process that is Golgi-localized γ-ear-containing Arf-binding protein (GGA) dependent, whereas insulin-stimulated translocation is regulated by Akt substrate of 160 kDa (AS160). In the present study, using a variety of GLUT4/GLUT1 chimeras, we have analyzed the specific motifs of GLUT4 that are important for GGA and AS160 regulation of GLUT4 trafficking. Substitution of the amino terminus and the large intracellular loop of GLUT4 into GLUT1 (chimera 1-441) fully recapitulated the basal state retention, insulin-stimulated translocation, and GGA and AS160 sensitivity of wild-type GLUT4 (GLUT4-WT). GLUT4 point mutation (GLUT4-F5A) resulted in loss of GLUT4 intracellular retention in the basal state when coexpressed with both wild-type GGA and AS160. Nevertheless, similar to GLUT4-WT, the insulin-stimulated plasma membrane localization of GLUT4-F5A was significantly inhibited by coexpression of dominant-interfering GGA. In addition, coexpression with a dominant-interfering AS160 (AS160-4P) abolished insulin-stimulated GLUT4-WT but not GLUT4-F5A translocation. GLUT4 endocytosis and intracellular sequestration also required both the amino terminus and large cytoplasmic loop of GLUT4. Furthermore, both the FQQI and the SLL motifs participate in the initial endocytosis from the plasma membrane; however, once internalized, unlike the FQQI motif, the SLL motif is not responsible for intracellular recycling of GLUT4 back to the specialized compartment. Together, we have demonstrated that the FQQI motif within the amino terminus of GLUT4 is essential for GLUT4 endocytosis and AS160-dependent intracellular retention but not for the GGA-dependent sorting of GLUT4 into the insulin-responsive storage compartment.

506 Background: Therapeutic drug monitoring (TDM) has been recognized as a useful tool for optimizing the dose of many drugs. The effectiveness of TDM for molecular targeted agents, however, has not so far been established. In this study, we examined the associations between pharmacokinetics of axitinib (AXI) and adverse events (AEs) and clinical outcomes in patients with advanced renal cell carcinoma (aRCC). Methods: The pharmacokinetics of AXI was examined in 24 patients (18 males and 6 females) with aRCC. The plasma AXI levels were measured using high-performance liquid chromatography and a pharmacokinetic study was performed on day 7 in the first cycle. Subsequently, the associations between the pharmacokinetics of AXI and the occurrence and grade of AEs, best tumor response, and progression-free survival (PFS) were investigated. Results: All the patients were been treated with 10 mg/day of AXI at the day of pharmacokinetics and the trough levels were ranged between 1.2 and 70.97 ng/mL. The trough levels were significantly associated with the occurrence of hypertension, hyperthyroidism, and grade 1 or higher proteinuria (p=0.037, 0.024, 0.027 by Mann-Whitney test, respectively), and were associated with the best tumor response (by Mann-Whitney test, p=0.043) (Table 1). Patients with 5.0 ng/mL of higher trough levels of AXI showed a tendency toward longer PFS than those with the trough levels less than 5.0 ng/mL (Table 2). Conclusions: TDM of AXI in patients with aRCC may be useful to determine the adequate dose and prevent severe AEs. Further studies are mandatory to conclude the usefulness of TDM of AXI for the long-term clinical efficacy in patients with aRCC. [Table: see text] [Table: see text]

Regulated transcription controls the diversity, developmental pathways and spatial organization of the hundreds of cell types that make up a mammal. Using single-molecule cDNA sequencing, we mapped transcription start sites (TSSs) and their usage in human and mouse primary cells, cell lines and tissues to produce a comprehensive overview of mammalian gene expression across the human body. We find that few genes are truly 'housekeeping', whereas many mammalian promoters are composite entities composed of several closely separated TSSs, with independent cell-type-specific expression profiles. TSSs specific to different cell types evolve at different rates, whereas promoters of broadly expressed genes are the most conserved. Promoter-based expression analysis reveals key transcription factors defining cell states and links them to binding-site motifs. The functions of identified novel transcripts can be predicted by coexpression and sample ontology enrichment analyses. The functional annotation of the mammalian genome 5 (FANTOM5) project provides comprehensive expression profiles and functional annotation of mammalian cell-type-specific transcriptomes with wide applications in biomedical research.

In previous studies, we showed that pretreatment of rat FRTL-5 thyroid cells with TSH, or other agents that increased intracellular cAMP, markedly potentiated DNA synthesis in response to insulin-like growth factor-I (IGF-I). In addition, we found that TSH pretreatment caused an increase in tyrosine phosphorylation of intracellular proteins including an unidentified 125-kDa protein that was well correlated with the TSH-potentiating effect on DNA synthesis induced by IGF-I. These results suggested that cAMP amplified IGF-I-dependent signals for cell growth through changes of cAMP-dependent tyrosine phosphorylation. The present studies were undertaken to determine how tyrosine kinase activation followed by an increase in tyrosine phosphorylation is required for cAMP-dependent potentiation of DNA synthesis induced by IGF-I in this cell line. First of all, we measured tyrosine kinase or protein-tyrosine phosphatase activities in the cell lysates by the in vitro assay. Chronic treatment with TSH or (Bu)2-cAMP stimulated tyrosine kinase activity in the particulate fraction and protein-tyrosine phosphatase activity in the soluble fraction, suggesting that tyrosine kinase plays more important roles for a cAMP-dependent increase in tyrosine phosphorylation of intracellular proteins. The increased tyrosine kinase activity was sensitive to genistein, a potent tyrosine kinase inhibitor. Genistein abolished both the cAMP-dependent increase in tyrosine phosphorylation of the 125-kDa protein and the enhanced DNA synthesis induced by IGF-I in a similar concentration-dependent manner. The only tyrosine-phosphorylated protein associated with the p85 regulatory subunit of phosphatidylinositol (PI) 3-kinase in response to cAMP was 125 kDa. In addition, we found that PI 3-kinase activity bound to p85 subunit significantly increased after (Bu)2cAMP treatment. These results suggested that cAMP stimulates PI 3-kinase through tyrosine phosphorylation of the 125-kDa protein. We then measured DNA synthesis in cells pretreated for 24 h with TSH or (Bu)2cAMP in the absence or presence of LY294002, a PI 3-kinase inhibitor, followed by treatment with IGF-I for 24 h. Presence of LY294002 during TSH or (Bu)2cAMP pretreatment completely abolished cAMP-dependent potentiation of DNA synthesis induced by IGF-I. These results suggest that in FRTL-5 cells cAMP activates genistein-sensitive tyrosine kinases that in turn activate PI 3-kinase activity. These mechanisms appear to be necessary for cAMP-dependent potentiation of the DNA synthesis induced by IGF-I.

A pot experiment was conducted with an Andosol to investigate the changes of phosphorus(P) compounds in the rhizosphere and hyphesphere soil (Organic P, Ca-P, Al-P, Fe-P). To distinguish between the effect of the arbuscular mycorrhizal root system and only of mycorrhizal hyphae, “a double pot system” (separation of a root and hyphae compartment by a nylon net) has been used. Ca-P and Al-P fraction around the mycorrhizal roots and mycorrhizal hyphae of both plant species, groundnut and pigeonpea significantly decreased under non-sterilized condition after 100 days of cultivation during the summer. This indicates that Ca-P and Al-P fractions in the Andosol with high accumulated P were quite mobile and available for these plants.

Estrogen deficiency in postmenopausal women is a major contributing factor to bone loss occurring after menopause. Although serum estradiol levels decrease conspicuously, a considerable amount of estrone still exists in women after natural menopause. Since most of the estrone is converted from androgens in adipose tissues, we attempted to seek interrelationship among serum estrone levels, fat mass and bone mineral density (BMD) in postmenopausal women. Thirty women who underwent natural menopause at the age between 45 and 53 were examined for BMD of head, arms, trunk and legs, as well as total body fat and total lean body mass using a Lunar DPX, a dual energy X-ray absorptiometer. Serum estrone concentration in each subject was measured by RIA. BMD of all sites of bones examined showed a positive correlation with relative fat mass (total body fat/body weight). Serum estrone levels were positively correlated with BMD of arms, trunk and legs, as well as relative fat mass. Multiple regression analysis further demonstrated that relative fat mass and serum estrone levels were significantly related to BMD of both arms and trunk, and BMD of trunk, respectively. These results suggest that estrone produced in adipose tissues may have, if only in part, a role in modulating the loss of bone mass occurring after menopause.

L. leichmanniiを用いた牛乳・調製粉乳中のビタミンB12の測定条件につき検討し,以下の結果を得た. (1) 接種用培地の前培養時間は5時間,接種菌液濃度はOD520=0.01,本培養時間は25時間,波長520nmでの測定が最適と思われた.測定時間は従来の32～44時間から30時間に短縮された. (2) 保存菌株の継代培養は毎日実施する必要はなく,1カ月間隔でも活性の低下は認められなかった.416日本食品工業学会誌 第36巻 第5号 1989年5月(60) (3) 新試験法により添加回収実験を試みたところ,良好な結果を得た. (4) 日内・日間分析精度を求めたところ従来法と比較して小さい値を得た.

It is considered that there were several possible pathways in which polypeptides were produced on premitive earth. Of these pathways, (i) condensation of solid amino acids on heating, (2) condensation of amino acids in aqueous media over condensing agents or (3) on heating are known. The first one has been studied in detail(Fox et al., 1960). The second one has not given longer peptides than 5 residues(Hulshof et al., 1976). However, there are a few reports related to the third one in which much larger peptides were obtained from aspargine with the molecular weight more than c.a. 3000(Kovacs et al., 1961.; Harada et al., 1978). In this paper, we wish to describe the role of aspargine and glutamine having 8 or y primary amide for polypeptide formation in the hypothetical premitive ocean. We carried out the heating of aqueous solution contaning aspargine and other amino acids(glycine, alanine, and valine) or peptides which have asparagine or glutamine residue in the C-terminal. Heatin @ reaction of aqueous solution of as~arqine and some other amino acids. Five mmol L-aspargine monohydrate(750mg) and other amino acids[0.5-2.5 mmol(Gly, Ala, Val)]were dissolved in 5 ml H20 , and the solutions sealed in vacuo in a glass tube were heated at 120°C for 2-200 hr. After the reactions, the reaction solution was analyzed by amino acid analyzer and remaining sample was loaded into a Sephadex G-15 column. The elution was carried out with 0.5 M-acetic acid to give white amorophous powder. The 32 hours' reaction mixtures gave muximum isolated yield(>90mg). The IR of this compound showed typical acidic polypeptide absorption(1710, 1650, 1540 cm-l). Averaged molecular weight of this peptide was estimated to be 1500-3000 by gel filtration. Molar composition of the amino acids contained in the copolyamino acids were almost pallarel to the first composition of the reaction mixtures, when the molar ratio of other amino acid to L-aspargine was lower than 50 %. Heating reactions of amino acid solutions without aspargine