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Metabolic Adjustments of Lactate Dehydrogenase Isozymes to the Environmental Temperature in Bluegill (Lepomis macrochirus) (환경온도에 대한 파랑볼우럭(Lepomis macrochirus) 젖산탈수소효소 동위효소들의 대사조절)

  • Ku, Bora;Cho, Sung Kyu;Yum, Jung Joo
    • Journal of Life Science
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    • v.26 no.10
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    • pp.1105-1112
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    • 2016
  • The aim of this study was to examine the metabolic adjustment of lactate dehydrogenase (EC 1.1.1.27, LDH) isozymes to the environmental temperature in bluegill (Lepomis macrochirus). This study included three groups of bluegill collected in April (group Ⅰ), May (group Ⅱ), and September (group Ⅲ). The LDH activities of skeletal muscle, heart, and brain tissues were higher in group Ⅲ than in groups Ⅰ and Ⅱ. The citrate synthase (EC 4.1.3.7, CS) activity was higher in skeletal muscle but lower in heart and brain tissues of group Ⅱ as compared to group Ⅰ. In contrast, the CS activity was lower in skeletal muscle and higher in heart and brain tissues in group Ⅲ than in group Ⅱ. Furthermore, the LDH/CS activity ratio was higher in the skeletal muscle and brain in group Ⅲ than in groups Ⅰ and Ⅱ. Accordingly, anaerobic metabolism was increased in group Ⅲ. LDH A4, A2B2, and B4 isozymes were expressed in skeletal muscle, heart, liver, and brain tissues. The LDH C hybrid was detected in brain tissue. The LDH A4 isozyme was successfully purified by affinity chromatography. The molecular weight of the purified LDH A4 isozyme was 136 kDa and its optimal pH for enzymatic activity was 8.0. The KmPYR values of LDH in skeletal muscle were 0.161-0.227 mM using pyruvate as a substrate. These kinetic properties of LDH in skeletal muscle are consistent with the fact that bluegill is a cold-adapted species. These results may be useful for predicting the habitat use of this fish.

Polymorphisms in Heat Shock Proteins A1B and A1L (HOM) as Risk Factors for Oesophageal Carcinoma in Northeast India

  • Saikia, Snigdha;Barooah, Prajjalendra;Bhattacharyya, Mallika;Deka, Manab;Goswami, Bhabadev;Sarma, Manash P;Medhi, Subhash
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.18
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    • pp.8227-8233
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    • 2016
  • Background: To investigate polymorphisms in heat shock proteins A1B and A1L (HOM) and associated risk of oesophageal carcinoma in Northeast India. Materials and Methods: The study includes oesophageal cancer (ECA) patients attending general outpatient department (OPD) and endoscopic unit of Gauhati Medical College. Patients were diagnosed based on endoscopic and histopathological findings. Genomic DNA was typed for HSPA1B1267 and HSPA1L2437 SNPs using the polymerase chain reaction with restriction fragment length polymorphisms. Results: A total of 78 cases and 100 age-sex matched healthy controls were included in the study with a male: female ratio of 5:3 and a mean age of $61.4{\pm}8.5years$. Clinico-pathological evaluation showed 84% had squamous cell carcinoma and 16% were adenocarcinoma. Dysphagia grades 4 (43.5%) and 5 (37.1%) were observed by endoscopic and hispathological evaluation. The frequency of genomic variation of A1B from wild type A/A to heterozygous A/G and mutant G/G showed a positive association [chi sq=19.9, p=<0.05] and the allelic frequency also showed a significant correlation [chi sq=10.3, with cases vs. controls, OR=0.32, $p{\leq}0.05$]. The genomic variation of A1L from wild T/T to heterozygous T/C and mutant C/C were found positively associated [chi sq=7.02, p<0.05] with development of ECA. While analyzing the allelic frequency, there was no significant association [chi sq=3.19, OR=0.49, p=0.07]. Among all the risk factors, betel quid [OR=9.79, Chi square=35.0, p<0.05], tobacco [OR=2.95, chi square=10.6, p<0.05], smoking [OR=3.23, chi square=10.1, p<0.05] demonstrated significant differences between consumers vs. non consumers regarding EC development. Alcohol did not show any significant association [OR=1.34, chi square=0.69, p=0.4] independently. Conclusions: It can be concluded that the present study provides marked evidence that polymorphisms of HSP70 A1B and HSP70 A1L genes are associated with the development of ECA in a population in Northeast India, A1B having a stronger influence. Betel quid consumption was found to be a highly significant risk factor, followed by smoking and tobacco chewing. Although alcohol was not a potent risk factor independently, alcohol consumption along with tobacco, smoking and betel nut was found to contribute to development of ECA.

Molecular Cloning and Expression of Candida antarctica lipase B in Corynebacterium genus

  • Gonzalez, Tamara;M'Barek, Hasna Nait;Gomaa, Ahmed E.;Hajjaj, Hassan;Zhen, Chen;Dehua, Liu
    • Microbiology and Biotechnology Letters
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    • v.47 no.4
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    • pp.546-554
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    • 2019
  • This study, for the first time, reports the functional expression of lipase B derived from the yeast Candida antarctica (CALB) in Corynebacterium strain using the Escherichia coli plasmid PK18. The CALB gene fragment encoding a 317-amino-acid protein was successfully obtained from the total RNA of C. antarctica. CALB was readily produced in the Corynebacterium strain without the use of induction methods described in previous studies. This demonstrated the extracellular production of CALB in the Corynebacterium strain. CALB produced in the Corynebacterium MB001 strain transformed with pEC-CALB recombinant plasmid exhibited maximum extracellular enzymatic activity and high substrate affinity. The optimal pH and temperature for the hydrolysis of 4-nitrophenyl laurate by CALB were 9.0 and 40℃, respectively. The enzyme was stable at pH 10.7 in the glycine-KOH buffer and functioned as an alkaline lipase. The CALB activity was inhibited in the presence of high concentration of Mg2+, which indicated that CALB is not a metalloenzyme. These properties are key for the industrial application of the enzyme.

Synthesis of Polyurethane Foam/Organonanoclay/Phosphates Composites and its Characterization (폴리우레탄폼/유기나노점토/포스페이트 복합체의 합성과 그 특성)

  • Park, Kyeong-Kyu;Lee, Sang-Ho
    • Elastomers and Composites
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    • v.46 no.4
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    • pp.343-351
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    • 2011
  • We prepared polyurethane foam/cloisite30B/phosphates composites and characterized their rise time, density, cell morphology, and thermal properties. The composites were synthesized with polyadipatediol-cloisite30B composite (f=2.0), polyether-polyol (f=4.6), polymeric 4,4-diphenyl methane diisocyanate (f=2.5), and D-580 (phenyl polyoxyalkenyl phosphate). As a blowing agent, cyclopentane and distilled water were used at various concentrations of D-580 from 0 to 2.81 wt%. The rise times of PUF/Closite30B/Phosphate composites blown with distilled water were faster than those blown with cyclopentane by 30%. The composites blown with cyclopentane had spherical-shape cells and the cell diameter was decreased with increasing D-580 wt%. While $T_g$ of the composites blown with cyclopentane linearly decreased with increasing the D-580 content, the $T_g$ of the composites blown with distilled water increased with the D-580 content. All PUF/Closite30B/Phosphate composites began to decompose from $250^{\circ}C$. The composites blown with cyclopentane showed the second thermal decomposition at temperatures higher than $500^{\circ}C$. The thermal stability of all composites increased with the D-580 content. The effect of D-580 on the thermal stability of the composites was measured higher at the composites blown with distilled water.

Characterization of Acid Phosphatase from Welsh Onion (파의 Acid Phosphatase의 특성)

  • Kim, Gi-Nahm;Kim, Suk-Ji;Kim, Seok-Hwan;Park, In-Shik
    • Korean Journal of Food Science and Technology
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    • v.28 no.4
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    • pp.663-667
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    • 1996
  • Acid phosphatase (EC 3.1.3.2) from Welsh onion was partially purified by Sephacryl S-200 gel filtration and CM-Sepharose CL-6B ion exchange chromatography. The optimum pH and temperature of acid phosphatase from green onion were pH 5.5 and $60^{\circ}C$, respectively. The enzyme was the most stable at pH 6.0 and unstable above pH 9.0. The activation energy of the enzyme was determined to be 4.86kcal/mole. The enzyme utilized p-nitrophenyl phosphate most as a best substrate among tested possible substrates, while 5'-GMP and 5'-IMP were poor substrates for the enzyme. $K_{m.app.}$ of the enzyme with p-nitrophenyl phosphate as a substrate was identified as 0.87mM. Among metal ions and inhibitors tested, $Cr^{+++},\;Zn^{++},\;Cu^{++}$, molybdate and metavanadate ions inhibited the enzyme reaction drastically.

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Biochemical Properties of Lactate Dehydrogenase Eye-Specific C4 Isozyme: Lepomis macrochirus and Micropterus salmoides (젖산탈수소효소 eye-specific C4 동위효소의 생화학적 특성: 파랑볼우럭(Lepomis macrochirus)과 큰입우럭(Micropterus salmoides))

  • Yum, Jung-Joo;Ku, Bo-Ra
    • Journal of Life Science
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    • v.22 no.2
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    • pp.209-219
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    • 2012
  • The properties of lactate dehydrogenase (LDH, EC 1.1.1.27) eye-specific $C_4$ isozyme were studied by polyacrylamide gel electrophoresis, Western blotting, immunoprecipitation, and enzyme kinetics. Furthermore, we proposed the optimal conditions for measuring the activity of LDH eye-specific $C_4$ isozyme. The isozymes were detected in the cytosol of eye tissues from Lepomis macrochirus and Micropterus salmoides and were more similar to the $A_4$ than the $B_4$ isozyme. LDH/CS in the eye tissue of L. macrochirus was increased in September, so the ratio of anaerobic metabolism was high. The electrophoretic patterns of mitochondrial LDH were similar to those of cytosolic LDH in the eye tissues of L. macrochirus and Micropterus salmoides. LDH eye-specific $C_4$ isozyme from eye tissue was purified by preparative native-PAGE. The activities of LDH eye-specific $C_4$ isozymes in L. macrochirus and M. salmoides were reduced at concentrations greater than 0.2 mM and 0.1 mM of pyruvate, respectively. These concentrations remained at 5.2% and 15.8% as a result of the inhibition by 10 mM of pyruvate, so the degree of inhibition was very high. The LDH activities of eye tissues were reduced at concentrations greater than 22 mM and 24 mM of lactate, respectively, in L. macrochirus and M. salmoides. The ${K_m}^{PYR}$ of eye-specific $C_4$ was 0.088 mM in L. macrochirus and it was 0.033 mM in M. salmoides. The activities of cytosolic and mitochondrial eye-specific $C_4$ isozymes were high in ${\alpha}$-ketobutyric acid. Furthermore, the activities of eye tissue and eye-specific $C_4$ isozyme had to be measured with 0.5 mM of pyruvate and a buffer solution of pH 7.5. As a conclusion, the eye-specific $C_4$ isozyme in M. salmoides has a high affinity for pyruvate and exhibits maximum activity at a lower concentration of pyruvate and at higher concentration of lactate than that in L. macrochirus. Therefore, it seems that the energy produced by the LDH eye-specific $C_4$ isozyme in M. salmoides was used at the first stage of predatory behavior.

Efficacy of Tomato Powder as Antioxidant in Cooked Pork Patties

  • Kim, I.S.;Jin, S.K.;Yang, M.R.;Chu, G.M.;Park, J.H.;Rashid, R.H.I.;Kim, J.Y.;Kang, S.N.
    • Asian-Australasian Journal of Animal Sciences
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    • v.26 no.9
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    • pp.1339-1346
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    • 2013
  • This study was carried out to determine the effects of tomato powder (TP) on cooked pork patties during storage at $10{\pm}1^{\circ}C$ in the dark. The total phenolic and flavonoid contents of TP extract were 26.22 mg gallic acid/100 g and 3.52 mg quercetin/100 g, respectively. The extract of TP showed a potential antioxidant activity in the DPPH radical-scavenging assay ($EC_{50}=16.76{\mu}g/mL$). Pork patties were manufactured with 0.25% (T1), 0.5% (T2), 0.75% (T3) and 1.0% (T4) TP in a basic formula (C). The pH and volatile basic nitrogen (VBN) values of T2, T3 and T4 patties were lower (p<0.05) than the C patties during storage. Increased concentration of TP in meat patties decreased (p<0.05) the 2-thiobarbituric acid reactive substances (TBARS) and total plate count (TPC) values at d 7 of storage. Tomato treated-patties had lower (p<0.05) values for lightness ($L^*$), but higher (p<0.05) values for redness ($a^*$) and yellowness ($b^*$) at d 3 and 7 of storage compared with the C. In the case of sensory evaluation, the scores of colour, flavour and overall acceptability of T3 and T4 patties were higher (p<0.05) than those of the C patty after 3 or 7 days of storage.

Parthenogenetic Activation and Development of Freshly Matured Bovine IVM Oocytes (체외성숙 직후 소 난포란의 단위발생과 체외발육능)

  • 정희태;임석기;오세훈;박춘근;양부근;김정익
    • Korean Journal of Animal Reproduction
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    • v.21 no.1
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    • pp.71-78
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    • 1997
  • This study was conducted to investigate the activation condition of freshly matured bovine IVM oocytes for use as a cytoplasmic recipient in nuclear transfer. Bovine oocytes matured in vitro for 22-24 h were treated with various activation conditions. In Experiment 1 in vitro matured oocytes were treated with electric stimuIus (ES; 2 pulses of 1.25 kV/cm for 70 ${\mu}{\textrm{s}}$ec, each pulse 1 sec apart), ethanol (ET; 7%, 5min) , Ca$^2+$-ionophore(A23187; 10$\mu\textrm{g}$/ml, 5min) and cycloheximide(CH; 10$\mu\textrm{g}$/ml, 6 h). Activation rates were similar in treatments with ES, ET and A23187(48.8~54.3%), however, significantly reduced with CH treatment(15.9%, P

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Developmental characteristics of Hemiptarsenus sp.(Hymenoptera : Eulophidae), a parasitoid of Liriomyza trifolii (Burgess) and effect of the insectcides (아메리카잎굴파리 기생봉, Hemiptarsenus sp.의 발육특성 및 살충제의 영향)

  • Moon, Hyung-Chol;Choi, Jeong-Sik;Hwang, Chang-Yeon
    • The Korean Journal of Pesticide Science
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    • v.4 no.4
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    • pp.72-76
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    • 2000
  • The study was conducted to investigate the developmental periods and effect of several insecticides on Hemiptarsenus sp., ectoparasitoid of Liriomyza trifolii. The mean length and width of egg were 0.5mm and 0.1mm. The mean length of larva, pupae, abult female, and abult male were 1.9mm, 2.0mm, 2.2mm, and 1.8mm, respectively. Developmental periods of Hemiptarsenus sp. from egg to larva at 15, 20, 25, $30^{\circ}C$ were 16.9, 8.8, 5.9, and 4.5 days, and those of pupa were 20.7, 9.7, 5.6, and 3.4 days, respectively. Based on these results, developmental threshold temperatures and effective temperatures were $9.5^{\circ}C$, 91.5 degree-days in egg-larval stage, $10.9^{\circ}C$, 142.3 degree-days in pupal stage. When several insecticides were evaluated to Hemiptarsenus sp. at the recommended concentrations, B.t WP, diflubenzuron WP, and cyromazin were negligiblly effective all life stages. Fipronil SC, cartap SP, spinosad GW were less toxic to larva and pupa, but highly toxic to adults. Abamectin EC was less toxic to all life stages, but inhibited oviposition of 50% more to Hemiptarsenus sp. female.

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In Vitro Antioxidant Activities and Antimicrobial Activity of Lotus (Leaf, Stem, and Seed Pod) Extracts (연잎, 연 줄기 및 연자방 추출물의 In Vitro 항산화 활성과 항균 활성에 관한 연구)

  • Lee, Ki-Won;Kim, Yong-Hwan;Shin, Kyung-Ok
    • The Korean Journal of Food And Nutrition
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    • v.30 no.4
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    • pp.771-779
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    • 2017
  • The purpose of this study was to investigate and analyze the total phenolic content (TPC), antioxidant activities (FRAP, ABTS, and DPPH), and antibacterial properties of lotus (Nelumbo nucifera) extracts. Lotus leaves, stems, and seed pods were extracted with deionized water at $95^{\circ}C$, and with 70.5% ethanol at $85^{\circ}C$. The TPC ranged from 8.12 to 215.12 GAE mg/g. The ethanol extract of the seed pod had the highest TPC, and the TPC of the corresponding deionized water extract was 161.45 mg/g. FRAP values ranged from 104.03 to $3,546.39TEAC\;{\mu}mol/g$, ABTS radical cation scavenging activities ranged from 105.11 to $3,956.94TEAC\;{\mu}mol/g$, and DPPH radical scavenging activities ranged from 37.29 to $2,549.46TEAC\;{\mu}mol/g$. $EC_{50}$ values ranged from 0.26 to 9.63 mg/mL, and 0.31 to 21.21 mg/mL for ABTS and DPPH, respectively. The ethanol and deionized water extracts of the seed pod showed higher TPC and stronger antioxidant properties (FRAP, ABTS, and DPPH) than those of characteristic of the leaf extracts. The ethanol and deionized water extracts of the seed pod showed antimicrobial activity against Bacillus subtilis, Staphylococcus aureus, and Pseudomonas aeruginosa with inhibition zones of 9.0 to 14.0 mm, and the ethanol extract of the leaf showed antimicrobial activity against B. subtilis and S. aureus with inhibition zones of 9.0 and 10.0 mm, respectively. Thus, the lotus seed pod could be used to produce novel teas, and could be a potential source of therapeutic ingredients for food and medicine.