• Title/Summary/Keyword: adenine nucleotide

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Effects of Tributyrin on Intestinal Energy Status, Antioxidative Capacity and Immune Response to Lipopolysaccharide Challenge in Broilers

  • Li, Jiaolong;Hou, Yongqing;Yi, Dan;Zhang, Jun;Wang, Lei;Qiu, Hongyi;Ding, Binying;Gong, Joshua
    • Asian-Australasian Journal of Animal Sciences
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    • v.28 no.12
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    • pp.1784-1793
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    • 2015
  • This study was carried out to investigate the effects of tributyrin (TB) on the growth performance, pro-inflammatory cytokines, intestinal morphology, energy status, disaccharidase activity, and antioxidative capacity of broilers challenged with lipopolysaccharide (LPS). A total of 160 one-day-old Cobb broilers were allocated to 1 of 4 treatments, with 4 replicated pens per treatment and 10 birds per pen. The experiment consisted of a $2{\times}2$ factorial arrangements of treatments with TB supplementation (0 or 500 mg/kg) and LPS challenge (0 or $500{\mu}g/kg$ body weight [BW]). On days 22, 24, and 26 of the trial, broilers received an intraperitoneal administration of $500{\mu}g/kg$ BW LPS or saline. Dietary TB showed no effect on growth performance. However, LPS challenge decreased the average daily gain of broilers from day 22 to day 26 of the trial. Dietary TB supplementation inhibited the increase of interleukin-$1{\beta}$ (in the jejunum and ileum), interleukin-6 (in the duodenum and jejunum), and prostaglandin $E_2$ (in the duodenum) of LPS-challenged broilers. Similar inhibitory effects of TB in the activities of total nitric oxide synthase (in the ileum) and inducible nitric oxide synthase (in the jejunum) were also observed in birds challenged with LPS. Additionally, TB supplementation mitigated the decrease of ileal adenosine triphosphate, adenosine diphosphate and total adenine nucleotide and the reduction of jejunal catalase activity induced by LPS. Taken together, these results suggest that the TB supplementation was able to reduce the release of pro-inflammatory cytokines and improve the energy status and anti-oxidative capacity in the small intestine of LPS-challenged broilers.

In vitro activity comparison of Erm proteins from Firmicutes and Actinobacteria (Firmicutes와 Actinobacteria에 속하는 세균들의 Erm 단백질 in vitro 활성 비교)

  • Jin, Hyung Jong
    • Korean Journal of Microbiology
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    • v.52 no.3
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    • pp.269-277
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    • 2016
  • Erm proteins methylate the specific adenine residue ($A_{2058}$, E. coli numbering) on 23S rRNA to confer the $MLS_B$ (macrolidelincosamide-streptogramin B) antibiotic resistance on a variety of microorganisms ranging from antibiotic producers to pathogens. When phylogenetic tree is constructed, two main clusters come out forming each cluster of Actinobacteria and Firmicutes. Two representative Erm proteins from each cluster were selected and their in vitro methylation activities were compared. ErmS and ErmE from Actinobacteria cluster exhibited much higher activities than ErmB and ErmC' from Firmicutes: 9 fold difference when ErmC' and ErmE were compared and 13 fold between ErmS and ErmB. Most of the difference was observed and presumed to be caused by N-terminal and C-terminal extra region from ErmS and ErmE, respectively because NT59TE in which N-terminal end 59 amino acids was truncated from wild type ErmS exhibited only 22.5% of wild type ErmS activity. Meanwhile, even NT59TE showed three and 2.2 times more activity when it was compared to ErmB and C, respectively, suggesting core region from antibiotic producers contains extra structure enabling higher activity. This is suggested to be possible through the extra region of 197RWS199 (from both ErmS and ErmE), 261GVGGSLY267 (from ErmS), and 261GVGGNIQ267 (from ErmE) and 291SVV293 (from ErmS) and 291GAV293 (from ErmE) by multiple sequence alignment.

Development of Selectable Marker of High Oleate Trait in Peanut (Arachis hypogaea L.) (땅콩에서 고 올레인산 형질관련 분자마커의 선발)

  • Yang, Kiwoung;Pae, Suk-Bok;Park, Chang-Hwan;Lee, Myoung Hee;Jung, Chan-Sik;Son, Jeong-Hee;Park, Keum-Yong
    • Korean Journal of Breeding Science
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    • v.42 no.5
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    • pp.507-514
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    • 2010
  • Peanut(Arachis hypogaea L.) is one of the major oilseed crops. The peanut oil consists of palmitic, oleic and linoleic acids, which are present at levels of 10%, 36-67% and 15-43%, respectively. High oleate mutant of peanut F435 contains 80% oleate and as little as 2% linoleate in seed oil. Previous study indicated that delta 12 fatty acid desaturase is a major enzyme controlling the oleate content in seeds of oilseed crops. F435 sequence alignment of their coding regions disclosed that an extra A(adenine) was inserted at the position +2,823 bp of delta 12 fatty acid desaturase gene. This study was to develop molecular marker (SNP marker) co-segregating with the high oleate trait. Chopyeong ${\times}$ F435 $F_2$ 41 population were investigated using molecular marker and fatty acid assay (NIR and gas chromatography). Finally, this marker segregates Chopyeong type 26 lines, heterotype 9 lines and F435 type 6 lines. These results in our study suggested that SNP marker conform fatty acid assay.

Etiological Properties and Coat Protein Gen Analysis of Potato Virus Y Occuring in Potatoes of Korea (우리나라 감자에 발생하는 PVY의 병원학적 특성 및 외피단백질 유전자 분석)

  • ;Richard M. Bostock
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 1995.06b
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    • pp.77-96
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    • 1995
  • To obtain basic informations for the improvement of seed potato production in Korea, some etiological properties of potato virus Y(PVY) distributed in the major seed potato production area(Daekwanryeong) were characterized, and the nucleotide and amino acid sequences of the coat protein gene of the PVY strains isolated were analyzed. PVY strains in Daekwonryeong, an alpine area, were identified to be two strains, PVYo and PVYN by symptoms of indicator plants, and their distribution in potato fields was similar. Major symptom on potato varieties by PVY was grouped as either mosaic alone or mosaic accompanied with veinal necrosis in the lower leaves. The symptom occurrence of the two symptoms was similar with Irish Cobbler, but Superior showed a higher rate of mosaic symptom than the other. The PVY strain which was isolated from potato cv. Superior showing typical mosaic symptoms produced symptoms of PVY-O on the indicator plants of Chenopodium amaranticolor, Nicotiana tabacum cv. Xanthi nc and Physalis floridana, but no symptom o Capsicum annum cv. Ace. Moreover, results from the enzyme-linked immunosorbent assay with monoclonal and polyclonal antibodies showed that the isolated PVY reacts strongly with PYV-O antibodies but does not react specifically with PVY-T antibodies. The purified virus particles were flexious with a size of 730$\times$11nm. On the basis of the above characteristics, the strain was identified to be a PVY-O and named as of PVY-K strain. The flight of vector aphids was observed in late May, however, the first occurrence of infected plants was in mid June with the bait plants surrounded with PVY-infected potato plants and early July with the bait plants surrounded with PVY-free potato plants. PVY infection rates by counting symptoms on bait plants (White Burley) were 1.1% with the field surrounded with PVY-free potato plants and 13.7% the fields surrounded with PVY-infected potato plants, showing the effect of infection pressure. The propagated PVY-K strain on tobacco(N. sylvestris) was purified, and the RNA of the virus was extracted by the method of phenol extraction. The size of PVY-K RNA was measured to be 9, 500 nucleotides on agarose gel electrophoresis. The double-stranded cDNAs of PVY-K coat protein(CP) gene derived by the method of polymerase chain reaction were transformed into the competent cells of E. coli JM 109, and 2 clones(pYK6 and pYK17) among 11 clones were confirmed to contain the full-length cDNA. Purified plasmids from pYK17 were cut with Sph I and Xba I were deleted with exonuclease III and were used for sequencing analysis. The PVY-K CP gene was comprised of 801 nucleotides when counted from the clevage site of CAG(Gln)-GCA(Ala) to the stop codon of TGA and encoded 267 amino acids. The molecular weight of the encoded polypeptides was calculated to be 34, 630 daltons. The base composition of the CP gene was 33.3% of adenine, 25.2% of guanine, 20.1% of cytosine and 21.4% of uracil. The polypeptide encoded by PVY-K CP gene was comprised of 22 alanines, 20 threonines, 19 glutamic acids and 18 glycines in order. The homology of nucleotide sequence of PVY-K CP gene with those of PVY-O(Japan), PVY-T(Japan), PVY-TH(Japan), PVYN(the Netherlands), and PVYN(France) was represented as 97.3%, 88.9%, 89.3%, 89.6% and 98.5%, respectively. The amino acid sequence homology of the polypeptide encoded by PVY-K CP gene with those encoded by viruses was represented as 97.4%, 92.5%, 92.9%, 92.9%, and 98.5%, respectively.

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Some Tertiary Interactions in 5S rRNA from Xanthomonas celebensis (Xanthomonas celebensis 5S rRNA의 몇 가지 삼차상호작용)

  • Bongrae Cho;Yeonghoon Lee;Myung-Un Choi;Inwon Park
    • Journal of the Korean Chemical Society
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    • v.37 no.2
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    • pp.237-243
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    • 1993
  • The primary and secondary structure of the 5S rRNA isolated from Xanthomonas celebensis were determined by enzymatic and chemical degradation methods. It consists of 119 nucleotides and contains no modified nucleosides. As with the 5S rRNAs of X. maltophilia and X. citri, it contains an additional uridine residue on the 5'-terminus. Its secondary structure was almost identical to the models previously proposed by us for the 5S rRNA of two Xanthomonas species. Its secondary structure consists of five helices, five loops and two bulges. The tertiary interactions in the 5S rRNA molecule were analyzed by Fe(II)-EDTA treatment and hybridization method using deoxyhexamer. From the fact that some adenine residues in loop M, region $I_1-C$, loop $H_1$, and loop $H_2$ become susceptible to diethylpyrocarbonate when the 5S rRNA was hybridized with deoxyhexamer complementary to the sequence $U_{35}CCCAU_{40}$ and that some nucleotide residues in loop M, loop $H_1$ and region $D-I_2$ become resistant Fe(II)-EDTA cleavage in the presence of $Mg^{2+}$, it is presumed that loops $H_1$ and $H_2$ interact with loop M in some way. In the tertiary interaction, the regions $I_1-C$ and $D-I_2$ seem to act as hinges in folding the stems $B-I_1-C$ and $D-I_2-E.$ It was found that loop $H_1$ changes into a smaller loop of three bases by forming noncanonical A : C base-pairs ih acidic environment.

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1828G>A polymorphism of the UDP-glucuronosyltransferase gene (UGT1A1) for neonatal hyperbilirubinemia in Koreans (한국인 신생아 황달에서 UGT1A1 유전자의 1828G>A 단일염기다형성에 관한 연구)

  • Kim, Ja Young;Kim, Mi Yeoun;Kim, Ji Sook;Kim, Eun Ryoung;Yoon, Seo Hyun;Lee, Hee Jae;Chung, Joo Ho
    • Clinical and Experimental Pediatrics
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    • v.49 no.1
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    • pp.34-39
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    • 2006
  • Purpose : The incidence of neonatal hyperbilirubinemia is twice as high in East Asians as in Caucasians. However, its metabolic basis has not been clearly explained. The UDP-glucuronosyltransferase gene(UGT1A1) mutation was found to be a risk factor of neonatal hyperbilirubinemia. We studied whether neonatal hyperbilirubinemia is associated with the 1828G>A(rs 10929303) polymorphism of the UGT1A1 gene, which encodes for a key enzyme of bilirubin metabolism. Methods : The genomic DNA was isolated from 80 Korean full term neonates who had greater than a 12 mg/dL level of serum bilirubin with no obvious cause, and the genomic DNA was also isolated from 164 Korean neonates of the control population. We studied a single nucleotide polymorphism (SNP) of 1828G>A in the untranslated region of the UGT1A1 gene by direct sequencing. Results : Three of the 80 neonates with a serum bilirubin level above 12 mg/dL had homozygous mutations and 10 of the 80 neonates with a serum bilirubin level above 12 mg/dL had heterozygous mutations. Thirteen of the 164 neonates of the control group had homozygous mutations and 16 neonates of the control group had heterozygous mutations. The allele frequency of 1828G>A polymorphism of UGT1A1 in the hyperbilirubinemia group was 10.0 percent, which was not significantly different from the allele frequency of 12.8 percent in the control group. Conclusion : In this study, the 1828G>A polymorphism of the UGT1A1 gene was detected in the Korean neonates with neonatal hyperbilirubinemia. Our results indicated that this SNP is not associated with the prevalence of hyperbilirubinemia in Koreans.

Studies on Degradation of Nucleic acid and Related Compounds by Microbial Enzymes (미생물 효소에 의한 핵산 및 그의 관련물질의 분해에 관한 연구)

  • Kim, Sang-Soon
    • Applied Biological Chemistry
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    • v.13 no.2
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    • pp.111-129
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    • 1970
  • As a series of studies on the nucleic acids and their related substances 210 samples were collected from 76 places such as farm soil, compost of heap, nuruk and meju to obtain microbial strains which produce 5'-phosphodiesterase. From these samples total of 758 strains were isolated by the use of dilution pour plate method. For all isolated strains primary screening of the productivity of RNA depolymerase was performed and useful strains with regard to 5'-phosphodiesterase productivities were identified. For these useful strains optimum condition, the effect of various compounds on the activity of 5'-phosphodiesterase, and the optimum condition for enzyme reaction were discussed. The quantitative of 5'-mononucleotides produced by the action of 5'-phosphodiesterase was performed using anion-exchange column chromatography and their identified was done by paper chromatography, thinlayer chromatography, ultra violet spectrophotometry, and characteristic color reaction using carbazole and schiff's reagent. (1) Penicillium citreo-viride PO 2-11 and Streptomyces aureus SOA 4-21 from soil were identified as a potent 5'-phosphodiesterase producing strains. (2) Optimum culture conditions for Penicillium citreo-viride PO 2-11 strain isolated were found to be pH 5.0 and $30^{\circ}C$, and the optimum conditions for enzyme action of 5'-phosphodiesterase were pH 4.2 and $60^{\circ}C$. Best carbon source for the production of 5'-phosphodiesterase was found to be sucrose and ammonium nitrate for nitrogen source. Addition of 0.01% corn steep liquor or yeast extract exhibited 20% increase in the amount of 5'-phosphodiesterase production compared to the control. 5'-phosphodiesterase produced by this strain was activated by $Mg^{++},\;Ca^{++},\;Zn^{++},\;Mn^{++}$ and was inhibited by EDTA, citrate, $Cu^{++},\;CO^{++}$. 5'-phosphodiesterase produced 5'-mononucleotide from RNA at a rate of 65.81%, and among the 5'-mononucleotides accumulated 5'-GMP only was found to have flavorous and the strain was also found lack of 5'-AMP deaminase. Productivity of flavorous 5'-GMP was found to be 186.7mg per gram of RNA. (3) Optimum culture canditions for the isolated Streptomyces aureus SOA 4-21 strain were pH 7.0 and $28^{\circ}C$, and the optimum conditions for the action of 5'-phosphodiesterase were pH 7.3 and $50^{\circ}C$. The best carbon source for 5'-phosphodiesterase production was found to be glucose and that of nitrogen was asparagine. Addition of 0.01% yeast extract exhibited increased productivity of 5'-phosphodiesterase by 40% compared to the non-added control. 5'-phosphodiesterase produced by this strain was activated by $Ca^{++},\;Zn^{++},\;Mn^{++}$ and was inhibited by citrate, EDTA, $Cu^{++}$. It was also found that the strain produce 5'-AMP deaminase in addition to 5'-phosphodiesterase. For this reason although decomposition rate was 63.58% the accumulation of 5'-AMP, 5'-CMP, 5'-GMP and 5'-UMP occurred by the breakdown of RNA. In the course of these reaction 5'-AMP deaminase converted 60% of 5'-AMP thus produced into 5'-IMP and flavorous 5'-mono nucleotide production was significantly increased by this strain over the above mentioned one. Production rates were found to be 171.8mg per grain of RNA for 5'-IMP and 148.2mg per gram of RNA for 5'-GMP, respectively.

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