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The rate-determining step in the dark state recovery process in the photocycle of PYP

  • Sasaki, Jun;Kumauchi, Masato;Hamada, Norio;Tokunaga, Fumio
    • Journal of Photoscience
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    • v.9 no.2
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    • pp.130-133
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    • 2002
  • The last step in the photocycle of photoactive yellow protein (PYP) is a spontaneous recovery of the dark state from the active state in which the p-coumaric acid chromophore is thermally isomerized, concomitantly with the deprotona- tion of the chtomophore and the refolding of the protein moicty. For the purpose of understanding the mechanism of the thermal back-isomerization, we have investigated the rate-determining step by analyzing mutant PYPs of Met100, which was previously shown to play a major role in facilitating the reaction (1). The mutation to Lys, Leu, Ala, or Glu decelerated the dark state recovery by 1 to 3 three orders of magnitude. By evaluating temperature-dependence and pH-dependence of the kinetics of the dark state recovery, it was found that the retardation by mutations resulted from elevation of the activation enthalpy ( H$\^$┿/) and that the pKa of the chromophore, which was affected by the mutation, is in a linier correlation with the amplitude of the rate constants. It was, therefore, deduced from the correlation that the free energy for crossing the activated state in the dark recovery process is proportional to the free energy for the deprotonation of the chromophore, identifying the rate-determining step as the deprotonation of the chromophore. (1) Devanathan, S. Genick, U. K. Canestrelli, I. L. Meyer, T. E. Cusanovich, M. A. Getzoff, E. D. Tollin, G., Biochemistry 1998, 37, 11563 - 11568

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Breeding of Productive Bivoltine Hybrid, CSR16${\times}$CSR17 of Silkworm Bombyx mori L.

  • Reddy, N.Mal;Basavaraja, H.K.;Kumar, N.Suresh;Joge, P.G.;Kalpana, G.V.;Dandin, S.B.;Datta, R.K.
    • International Journal of Industrial Entomology and Biomaterials
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    • v.8 no.2
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    • pp.129-133
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    • 2004
  • The breeding work was initiated by utilizing two Japanese hybrids namely C135${\times}$N134 and N137${\times}$C146 along with two evolved Indian breeds, J14 and A24. The breed CSR16 which is characterized with marked larvae and white dumbbell cocoons was evolved from the Japanese hybrid C135${\times}$N134 by crossing with J14, while the breed CSR17 which is characterized with plain larvae and white oval cocoons was evolved from the Japanese hybrid N137${\times}$C146 by crossing with A24. The hybrid of these evolved breeds i.e., CSR16${\times}$CSR17 has shown superior over control hybrid $KA{\times}{NB_4}{D_2}$ and on par with the ruling hybrid of CSR2${\times}$CSR4. The hybrid CSR16${\times}$CSR17 was authorized during 1999 by Central Silk Board, Bangalore, Government of India for commercial exploitation during favourable months based on national level race authorization test.

Immunological Studies on Oriental Pharmaceutical Preparations (I) (한약제제(漢藥製劑)의 면역학적(免疫學的) 연구(硏究) (제 1 보)(第 1 報))

  • Kim, Byong-Kak;Chung, Kyeong-Soo;Choi, Eung-Chil;Kim, Hye-Ryung;Park, Hee-Ju;Cho, Pil-Hyoung
    • Korean Journal of Pharmacognosy
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    • v.14 no.3
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    • pp.125-129
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    • 1983
  • To examine effects of oriental pharmaceutical preparations on immunity, four of such preparations which are currently being used were examined. After their fluid extracts were injected intraperitoneally to ICR mice, the counts of total cell number, PMN and macrophage in the peritoneal cavity were made by using non-specific esterase staining method, and plaque-forming cells (PFC) were measured by using the modified method of Cunningham and Jerne. The results showed that 'Bo-Jung-Ik-Ki-Tang', 'Ih-Jung-Tang', 'Ohn-Kyeong-Tang' and 'Ke-Ji-Bok-Ryeong-Hwan-Ka-Dae-Hwang' increased both the macrophages and the total cells of the peritoneal cavity and that these preparations increased the number of PFC in the spleen. Therefore the data indicate that they potentiate both of the cell­mediated and humoral immunities in the mice.

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Cloning and Sequencing of the ${\alpha}-1{\rightarrow}6$ Dextransurcrase Gene from Leuconostoc mensenteroides B-742CB

  • Kim, Ho-Sang;Kim, Do-Man;Ryu, Hwa-Ja;Robyt, John-F.
    • Journal of Microbiology and Biotechnology
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    • v.10 no.4
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    • pp.559-563
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    • 2000
  • A dextransucrase gene (dsrB742) that expresses a dextransucrase to synthesize mostly ${\alpha}-1{\rightarrow}6$ linked dextran with a low amount (3-5%) of ${\alpha}-1{\rightarrow}3$ branching was cloned and sequenced from Leuconostoc mesenteroides B-742CB. The 6.1-kb PstI fragments were ligated with pGEM-3Zf(-) and transformed into E. coli $DH5{\alpha}$. The recombinant clone (pDSRB742) synthesized dextran on an agar plate containing 2% (w/v) sucrose. The dextran synthesized was hydrolyzed with Penicillium endo-dextranase. The hydrolyzate was composed of glucose, isomaltose, isomaltotriose, and branced pentasaccharide. The nucleotide sequence of dsrB742 showed one open reading frame (ORF) composed of 4,524 bp encoding dextrasnsucrase. The deduced amino acid sequence revealed a calculated molecular mass of 168.6 kDa. It also showed an activity band of 184 kKa on a non-denaturing SDS-PAGE (10%). The amino acid sequence of DSRB742 exhibited a 50% similarity with DSRA from L. mesenteroides B-1299, a 70% similarity with DSRS from L. mesenteroides B-512 (F, FMCM) and a 45-56% similarity with Streptococcal GTFs.

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Cell Death and Proliferation after Treatment and Reinfection of Clonorchis sinensis in the Sprague-Dawley Rat Bile Duct

  • Min, Byoung-Hoon;Ahn, Ka-Young;Lee, Haeng-Sook;Kim, Soo-Jin;Joo, Kyoung-Hwan
    • Applied Microscopy
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    • v.45 no.2
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    • pp.80-88
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    • 2015
  • The structural change and distribution of mitochondrial enzyme (ATPase, cytochrome-c-oxidase), cell proliferation (proliferating cell nuclear antigen, PCNA), cell death (caspase-3) and cell growth factor (fibroblast growth factor 8, FGF-8) in the Sprague-Dawley rat bile duct during Clonorchis sinensis infection was investigated. Experimental groups were divided into C. sinensis infection, superinfection and reinfection of C. sinensis after 'praziquantel' treatment group. As a result, C. sinensis infected rat bile ducts showed the features of chronic clonorchiasis, i.e., connective tissue thickening, ductal fibrosis and epithelial tissue dilatation. PCNA for cell proliferation increased in the infection group, and decreased after praziquantel treatment. Caspase-3 was distributed in reinfection group only. FGF-8 was distributed in the rat bile duct after praziquantel treatment but not distributed in infection and reinfection group. Overall, C. sinensis infection causes physical and chemical irritations and then brings on the abnormalities of intracellular energy metabolism and cellular growth factors, which hinders bile duct tissue from functioning properly, and resultingly, fibrosis occurs and epithelial cells dilated abnormally. More intense infection makes tissue fibrosis chronical and activates apoptosis factors.

Comparison of Halloysitum Rubrum before and after Physical and Chemical Manipulation (적석지(赤石脂)의 포제전후의 성분 비교연구)

  • Kook, Yoon-Bum
    • The Korea Journal of Herbology
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    • v.20 no.2
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    • pp.17-26
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    • 2005
  • Objectives : The study has been undertaken to scrutinize the results of Halloysitum Rubrum before and after physical and chemical manipulation. Methods : I used X-rays diffracted analysis system(Phillips X'Pert - MPD System) on condition of 40Kv, Anode of Cu Ka to grasp the compositions of crude Halloysitum Rubrums which have been from here and there after pulverization them below 200 mesh. X-rays fluorescence analysis system have been used to check the chief elements content of the ore on condition of 40Kv, 95mA. ICP-MS analysis system(Perkin Elmer, Elan 6100)have been used after a micro-elements and a heavy metals take advantage of mixed acid were dissolved completely. Results : The compositions of the ore Halloysitum Rubrum are composed of large amount of Halloysite and Kaolinite, and small amount of Hematite, Alum, quartzite. The chief elements content of the ore Halloysitum Rubrum are composed of Al and Si. A heavy metals in Halloysitum Rubrum are As, Cd, Cr, Cu, Pb, Zn, etc. As and Zn's amounts was lessen after physical and chemical manipulation Halloysitum Rubrum are decreased at $400^{\circ}C\;and\;450-500^{\circ}C$ each by heat analysis system. Conclusion : Results of this study show that Halloysitum Rubrum can be used in a diverse ways to treat many kinds of disease.

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Introduction to Distribution and Ecology of Sterile Conks of Inonotus obliquus

  • Lee, Min-Woong;Hur, Hyeon;Chang, Kwang-Choon;Lee, Tae-Soo;Ka, Kang-Hyeon;Jankovsky, L.
    • Mycobiology
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    • v.36 no.4
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    • pp.199-202
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    • 2008
  • Inonotus obliquus is a fungus that causes white heart rot on several broad-leaved species. This fungus forms typical charcoal-black, sterile conks (chaga) or cinder conks on infected stems of the birche (Betula spp). The dark brown pulp of the sterile conk is formed by a pure mycelial mass of fungus. Chaga are a folk remedy in Russia, reflecting the circumboreal distribution of I. obliquus in boreal forest ecosystems on Betula spp. and in meridional mountain forests on beech (Fagus spp.) in Russia, Scandinavia, Central Europe, and Eastern Europe. Distribution at lower latitudes in Western and Southern Europe, Northern America, Asia, Japan, and Korea is rare. Infected trees grow for many years without several symptoms of decline. The infection can penetrate through stem injuries with exterior sterile conks developing later. In the Czech Republic, cinder conk is found on birches inhabiting peat bogs and in mountain areas with a colder and more humid climate, although it is widespread in other broad leaved species over the Czech Republic. The most common hosts are B. pendula, B. pubescens, B. carpatica, and F. sylvatica. Less frequent hosts include Acer campestre, Acer pseudoplatanus, Alnus glutinosa, Alnus incana, Fraxinus excelsior, Quercus cerris, Q. petraea, Q. robur, Q. delachampii, and Ulmus sp.

Cloning and Sequence Analysis of Two Catechol-degrading Gene Clusters from a Phenol-utilizing Bacterium Pseudomonas putida SM25

  • Jung, Young-Hee;Ka, Jong-Ok;Cheon, Choong-Ⅰll;Lee, Myeong-Sok;Song, Eun-Sook;Daeho Cho;Park, Sang-Ho;Ha, Kwon-Soo;Park, Young-Mok
    • Journal of Microbiology
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    • v.41 no.2
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    • pp.102-108
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    • 2003
  • A 6.1 kb Sph I fragment from the genomic DNA of Pseudomonas putida SM 25 was cloned into the veetor pUC19. The open reading frame of catB was found to consist of 1,122 nucleotides. The sequence alignment of the catB gene products from different kinds of bacteria revealed an overall identity ranging from 40 to 98%. The catC gene contained an open reading frame of 96 codons, from which a protein with a molecular mass of about 10.6 kDa was predicted. The amino acids in the proposed activesite region of CatC were found to be almost conserved, including the charged residues. Since the catBC genes in P. putida SM25 were tightly linked, the could be regulated under coordinate transcription, and transcribed from a single promoter located upstream of the catB gene, as in P. putida RBI.

Sex-Role Development in Late Childhood and Adolescence: -Relationships among Sex-Role Stereotypes, Sex-Role Ideology, Sex-Role Identity, and Self-Esteem (아동 후기에서 청소년기에 이르는 성역할발달에 관한 연구 - 성역할 고정관념, 성역할 이데올로기, 성역할 정체감, 자아존중감간의 관계를 중심으로 -)

  • Yoo, Ka Hyo
    • Korean Journal of Child Studies
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    • v.15 no.1
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    • pp.127-144
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    • 1994
  • The purpose of this study was to identify intrapsychic processes of self socialization in sex role development during late childhood and adolescence. For this purpose I investigated the sex-role stereotypes, sex-role ideology, sex-role identity, and self-esteem of 6th, 8th, and 10th graders and examined the causal relationships among these sex-role variables. Data were gathered through questionnaires administered twice with an interval of three months. The methods of analysis were one-way ANOVA, $x^2$, and multiple regression. The results showed, (1) Adolescents' sex-role stereotypes had significant relationships with sex typing. Subjects with low scores on sex-role stereotypes were more likely to show opposite sex typing. (2) Sex-role ideologies had significant relationships with sex typing. As Subjects agreed more with egalitarianism, they were more likely to exhibit opposite sex typing. (3) Adolescents' sex typing had significant relationships with their self-esteem. The androgynous group exhibited the highest scores on self-esteem. (4) Path analysis from the multiple regression analysis indicated different processes between sexes in sex-role development. In the boys' sex-role development, it was found that only masculinity contributed to self -esteem. Girls' masculinity also contributed most to self -esteem, but other sex -role variables such as femininity and sex-role stereotypes made some contributions to girls' self-esteem.

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Bioavailability Evaluation of Two Ceftriaxone Formulations Using Two Way Crossover Design in Voluteers

  • Kwon, Kwang-Il;Bok, Soo-Jin;Kim, Moo-Heon;Lee, Chang-Hyun
    • Archives of Pharmacal Research
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    • v.18 no.4
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    • pp.237-242
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    • 1995
  • For the vioequivalence study of two ceftriaxone injection formulations ($Rocephin{\circledR}$ ; Roche, and Triaxone ; Hanmi0, the HPLC analytical method for the analysis of ceftriaxone in plasma was used. Fourteen healthy volunteers completed the study and each subject were IM in jected signle doses (1 g) of the test and the reference formulations in a two-way crossover design with an one week drug free interval between doses. Following each administration, plasma concentrations of ceftrixone were monitored over a period of 24 h. Bioequivalence parameters $AUC_{24th}, {\;}T_{max}, {\;}C_{max}$ and MRT determined from the data obtained for the two formulations were examined by analyses of variance (ANOVA) and other criteria and tests for bioequivalence. Results of ANOVA and confidence limits of test/reference ratios of $AUC_{24th}, {\;}T_{max}, {\;}C_{max}$ and MRT, and statistical tests indicated the bioequivalence of the two formulations (i.e., test/reference ratio was within $100{\pm}20%$) except for $T_{max}$ The mean of $T_{max}$ showed only 6. 9% difference from the reference but the detection limit was 22.5% which is slightly over the 20% criteria. No pharmacokinetic parameters including Ka, Kel, Vd and Cl indicated significant difference in between the two fomulations. It was concluded that the data yielded fro the two cefriaxone formulations demonstrated that they were bioequivalent.

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