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Molecular Cloning and Functional Analysis of the Gene Encoding 3-hydroxy-3-methylglutaryl Coenzyme A Reductase from Hazel (Corylus avellana L. Gasaway)

  • Wang, Yechun;Guo, Binhui;Zhang, Fei;Yao, Hongyan;Miao, Zhiqi;Tang, Kexuan
    • BMB Reports
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    • v.40 no.6
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    • pp.861-869
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    • 2007
  • The enzyme 3-hydroxy-3-methylglutaryl-CoA reductase (HMGR; EC1.1.1.34) catalyzes the first committed step of isoprenoids biosynthesis in MVA pathway. Here we report for the first time the cloning and characterization of a full-length cDNA encoding HMGR (designated as CgHMGR, GenBank accession number EF206343) from hazel (Corylus avellana L. Gasaway), a taxol-producing plant species. The full-length cDNA of CgHMGR was 2064 bp containing a 1704-bp ORF encoding 567 amino acids. Bioinformatic analyses revealed that the deduced CgHMGR had extensive homology with other plant HMGRs and contained two transmembrane domains and a catalytic domain. The predicted 3-D model of CgHMGR had a typical spatial structure of HMGRs. Southern blot analysis indicated that CgHMGR belonged to a small gene family. Expression analysis revealed that CgHMGR expressed high in roots, and low in leaves and stems, and the expression of CgHMGR could be up-regulated by methyl jasmonate (MeJA). The functional color assay in Escherichia coli showed that CgHMGR could accelerate the biosynthesis of $\beta$-carotene, indicating that CgHMGR encoded a functional protein. The cloning, characterization and functional analysis of CgHMGR gene will enable us to further understand the role of CgHMGR involved in taxol biosynthetic pathway in C. avellana at molecular level.

Change of Gait Pattern of Patients with Ataxic Gait by Cerebellar Infarction in Comparison to Corticospinal Tract Stroke Patients (소뇌경색 환자의 운동기능 회복에 따른 보행 척도의 변화 관찰 연구 : 피질척수로 손상환자와의 비교를 중심으로)

  • Chu, Hongmin;Lee, Young-ung;Kim, Kwang-ho;Lim, Hyeon-seo;Ryu, Ho-sun;Park, Shin-hyeok;Kim, Cheol-hyun;Sung, Kang-keyng;Lee, Sang-kwan
    • The Journal of Internal Korean Medicine
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    • v.41 no.2
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    • pp.194-203
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    • 2020
  • Objectives: This study aimed to compare the gait patterns of cerebellar infarction patients with those of corticospinal tract stroke patients through a follow-up of patients with ataxic gait due to cerebellar infarction and corticospinal tract stroke. Methods: We investigated two cases of patients with cerebellar infarction and two cases each of acute or chronic corticospinal tract stroke who were hospitalized at Wonkwang University Gawangju Medical Center from September 1, 2017 to February 15, 2020 based on medical records and gait analyses. The spatiotemporal gait parameters of each patient were measured three times at 2-week intervals except those of the chronic corticospinal tract stroke patients, which were measured twice at a 1-month interval. Results: Spatiotemporal gait parameters, which include velocity, cadence, step length, stride length, and single support, were consistently increased in the cerebellar infarction patients in comparison to the corticospinal tract stroke patients. The stance phase was decreased in all the patients. Conclusions: The cerebellar infarction patients' gait spatiotemporal parameters were found to consistently improve. Moreover, gait analysis can be used to effectively measure improvement of ataxic gait.

Molecular Cloning and Function Analysis of an Anthocyanidin Synthase Gene from Ginkgo biloba, and Its Expression in Abiotic Stress Responses

  • Xu, Feng;Cheng, Hua;Cai, Rong;Li, Lin Ling;Chang, Jie;Zhu, Jun;Zhang, Feng Xia;Chen, Liu Ji;Wang, Yan;Cheng, Shu Han;Cheng, Shui Yuan
    • Molecules and Cells
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    • v.26 no.6
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    • pp.536-547
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    • 2008
  • Anthocyanidin synthase (ANS, leucoanthocyanidin oxygenase), a 2-oxoglutarate iron-dependent oxygenase, catalyzed the penultimate step in the biosynthesis of the anthocyanin class of flavonoids, from the colorless leucoanthocyanidins to the colored anthocyanidins. The full-length cDNA and genomic DNA sequences of ANS gene (designated as GbANS) were isolated from Ginkgo biloba for the first time. The full-length cDNA of GbANS contained a 1062-bp open reading frame (ORF) encoding a 354-amino-acid protein. The genomic DNA analysis showed that GbANS gene had three exons and two introns. The deduced GbANS protein showed high identities to other plant ANSs. The conserved amino acids (H-X-D) ligating ferrous iron and residues (R-X-S) participating in 2-oxoglutarate binding were found in GbANS at the similar positions like other ANSs. Southern blot analysis indicated that GbANS belonged to a multi-gene family. The expression analysis by real-time PCR showed that GbANS expressed in a tissue-specific manner in G. biloba. GbANS was also found to be up-regulated by all of the six tested abiotic stresses, UV-B, abscisic acid, sucrose, salicylic acid, cold and ethylene, consistent with the promoter region analysis of GbANS. The recombinant protein was successfully expressed in E. coli strain with pET-28a vector. The in vitro enzyme activity assay by HPLC indicated that recombinant GbANS protein could catalyze the formation the cyanidin from leucocyanidin and conversion of dihydroquercetin to quercetin, suggesting GbANS is a bifunctional enzyme within the anthocyanidin and flavonol biosynthetic pathway.

Accuracy of Electronic Pedometers to Assess Body Fatness in Obese Children and Youth (비만 어린이와 청소년들의 체지방 평가를 위한 electronic pedometer 의 정확성 분석)

  • Kim, Do-Yeon
    • Journal of Life Science
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    • v.19 no.10
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    • pp.1368-1373
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    • 2009
  • The purpose of this study was to assess the influence of waist size on the reliability and validity of pedometers to count steps in children and youth. The participants for this study were 20 children and youth, composed of 14 Hispanic and 6 Caucasian children. Ten children and youth had waist circumferences greater than the $85^{th}$ percentile (Body Mass Index (BMI)=$28.91\pm3.07$), and 10 children and youth had waist circumferences smaller than the $50^{th}$ percentile (BMI=$18.05\pm1.55$). To examine pedometer reliability, each child completed 3 ascent and descent trials up a set of 15 stairs while wearing a Yamax SW-701 pedometer. The main effect of trials was not statistically significant for the stair ascent trials F (2, 36)=2.575 or for the descent trials F (2, 36)=0.235. The trial by group interaction was also not statistically significant. To examine the influence of waist circumference on the validity of the pedometer in counting walking steps at a self-selected walking pace, the children and youth in the two groups completed a 400-m course. The main effect on the groups was statistically significant, F (1, 18)=7.489. The main effect of counting techniques was not statistically significant, F (1, 18)=2.983 (hand-counted vs. pedometer counted). Overall, the trial and trial by group interaction comparisons for the 400-m walk were not statistically significant, suggesting that the pedometer was equally valid as a tool for assessing walking steps in high waist circumference (HWC) and low waist circumference (LWC) in children and youth.

Detection Characteristics of TL, ESR and DNA Comet for Irradiated Soybeans (열발광, 전자스핀공명 및 DNA Comet 분석에 의한 대두의 방사선 조사 여부 검지 특성)

  • Lee, Eun-Young;Jeong, Jae-Young;Noh, Jung-Eun;Jo, Deok-Jo;Kwon, Joong-Ho
    • Korean Journal of Food Science and Technology
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    • v.34 no.1
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    • pp.18-23
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    • 2002
  • The detection characteristics of gamma-irradiated $(0{\sim}4\;kGy)$ soybeans produced in Korea and China were investigated by thermoluminescene (TL), electron spin resonance (ESR), and DNA comet assay. The TL glow curves were shown at around $200^{\circ}C$ for irradiated soybeans, while that at $280^{\circ}C$ for the non-irradiated one. The normalization with a re-irradiation step at 1 kGy could verify the above detection results. The Korean soybean showed higher glow curves than Chinese did. The ESR spectroscopy for husks of irradiated soybeans revealed specific signals (g = 2.02374, 1.98715) derived from cellulose radical, which intensities were proportional to irradiation does, with the higher peaks in Chinese sample than Korean one. The DNA comet for the non-irradiated sample showed no or little tails, while those for irradiated samples above 0.5 kGy were remarkably changed in their length, size, and concentration, thus resulting in distinguishing non-irradiated from irradiated samples. As a result, TL, ESR, and DNA comet determinations were found suitable for the detection of irradiated soybean at 0.5 kGy or more, and negligible differences were observed between Korean and Chinese origins in their detection characteristics.

Kinematic Analysis of Women's 100-m Final during IAAF World Championships, Daegu 2011 (2011 대구세계육상선수권대회 100 m 여자 결승전의 운동학적 분석)

  • Ryu, Ji-Seon;Ryu, Jae-Kyun;Kim, Tae-Sam;Park, Young-Jin;Hwang, Won-Seob;Yoon, Suk-Hoon;Park, Sang-Kyoon
    • Korean Journal of Applied Biomechanics
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    • v.21 no.5
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    • pp.521-528
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    • 2011
  • The purpose of this study was to analyze the kinematic characteristics of the finalists in the women's 100 m event to provide important information to coaches and athletes. Three different biomechanics techniques were applied for analyzing sprinter motion: LAVEG, a panning technique, and 12 video cameras for 3 dimensional analysis of the 40 m - 70 m portion of the race. Carmelita Jeter(USA) performed the maximum speed of 10.54 m/s at the distance of 58.2 m. There was a tendency to show a better performance time with a high number of steps (p=.13) and shorter stride length (p=.14) among the 8 sprints. Furthermore, the stride frequency and the performance time were negatively correlated as a higher stride frequency had a positive impact on the performance time (p=.02). Based on 3 dimensional analysis, the 4 top ranked sprinters used the different strategies to maintain a high COM (Center of Mass) velocity during the mid portion of the race (40 m - 70 m). Carmelita Jeter(USA) showed more flexed knee and hip motion at heel contact (HC) to maintain a high COM velocity while S.A. Fraser-Pryce (JAM) showed more extended knee and hip motion at HC. On the other hands, Veronica Campbell-Brown (JAM) and Kelly-Ann Baptiste (TRI) showed a tendency to have high knee lifts during the swing phase to maintain the high COM velocity during the race. These biomechanical analyses of the women's 100 m final event in the 2011 WC, Daegu, will provide important scientific information to coaches and athletes for understanding the sprinting mechanism of today's top-class sprinters.

Polymorphism of Bovine Lymphocyte Antigen DRB3.2 Alleles in Iranian Native Sarabi Cows

  • Pashmi, M.;Ghorashi, S.A.;Salehi, A.R.;Moini, M.;Javanmard, A.;Qanbari, S.;Yadranji-Aghdam, S.
    • Asian-Australasian Journal of Animal Sciences
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    • v.19 no.6
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    • pp.775-778
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    • 2006
  • Sarabi cows (n = 136) from the Sarabi Breeding Station were genotyped at bovine lymphocyte antigen (BoLA)-DRB3.2 locus by a genotyping system that used the polymerase chain reaction and restriction fragment length polymorphism. Genomic DNA was extracted from whole blood samples. A two-step polymerase chain reaction was carried out in order to amplify a 284 base-pair fragment of target gene. Nested-PCR products were digested with three restriction endonuclease enzymes RsaI, BstYI and HaeIII. Digested fragments were analyzed by polyacrylamide gel electrophoresis. Twenty-six BoLA-DRB3.2 alleles were identified with frequencies ranging from 0.4 to 15.1%. Six new allele types observed in this study have not been reported previously. Identified alleles include: BoLA-DRB3.$2^*1$, $^*2$, $^*4$, $^*6$, $^*8$, $^*12$, $^*13$, $^*14$, $^*15$, $^*16$, $^*17$, $^*23$, $^*24$, $^*25$, $^*28$, $^*32$, $^*34$, $^*35$, $^*36$, $^*37$, $^*42$, $^*46$, $^*51$, $^*kba$, $^*laa$ and $^*vaa$. Their frequencies were found to be 0.4, 0.4, 0.7, 11.4, 1.1, 1.8, 2.9, 2.2, 4.4, 9.6, 1.1, 13.6, 0.4, 0.4, 1.1, 0.7, 0.4, 6.2, 2.2, 3.7, 1.1, 7.7, 1.5, 15.1, 2.6 and 7.3% respectively. The six most frequent alleles (DRB3.2 $^*6$, $^*16$, $^*23$, $^*46$, $^*kba$ and $^*vaa$) accounted for 64.7% of the alleles in the population of this herd. Numerous studies on this locus, covering different breeds, has revealed the existence of various alleles in this locus, and new investigations have introduced novel alleles. With respect to the high number of the observed alleles in this survey and the novelty of some alleles with no previous record of reporting, it is plausible to conclude that the BoLA-DRB3.2 locus is highly polymorphic in Iranian native Sarabi cows.

Manufacture of Printing and Writing Papers from Old Corrugated Containers (OCC) (Old Corrugated Containers (OCC)로부터 인쇄·필기용지 제조)

  • Lee, Goo;Ahn, Byoung-Jun;Paik, Ki-Hyon
    • Applied Chemistry for Engineering
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    • v.10 no.3
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    • pp.367-373
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    • 1999
  • Increasing consumer demand for recycle of most paper products is challenging manufacturers to use lower value raw materials while maintaining high quality. This study was carried out to investigate the composition of old corrugated containers (OCC) and to manufacture printing and writing papers of over 85% (ISO) brightness by ECF and TCF bleaching from OCC. Fiber length of American old corrugated container (AOCC) was longer than that of Korean old corrugated container (KOCC) in fiber length. The former was composed of unbleached kraft pulp (UKP) of about 80 percentage, the latter about 20 percentage. OCC is feasible for manufacturing of pulps i.e., AOCC is for printing and writing papers and KOCC is for whiteboards, in the aspects of brightness and mechanical properties on the other hand, the cost of manufacturing upgraded papers by OCC is high because of bleaching chemicals. Neverthless, considering various aspects, especially environment, there are many advantages in recycling. In case of the treated AOCC pulp, over 85% (ISO) brightness can be achieved through TCF or ECF bleaching step, while the treated KOCC pulp did not allow high brightness through TCF bleaching. The tensile index of the bleached KOCC pulp was lower than that of the bleached AOCC pulp, but the burst and tear index of KOCC was higher than that of AOCC.

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Unsteady 2-D flow field characteristics for perforated plates with a splitter

  • Yaragal, Subhash C.
    • Wind and Structures
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    • v.7 no.5
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    • pp.317-332
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    • 2004
  • Wind tunnel experiments were conducted under highly turbulent and disturbed flow conditions over a solid/perforated plate with a long splitter plate in its plane of symmetry. The effect of varied level of perforation of the normal plate on fluctuating velocities and fluctuating pressures measured across and along the separation bubble was studied. The different perforation levels of the normal plate; that is 0%, 10%, 20%, 30%, 40% and 50% are studied. The Reynolds number based on step height was varied from $4{\times}10^3$ to $1.2{\times}10^4$. The shape and size of the bubble vary with different perforation level of the normal plate that is to say the bubble is reduced both in height and length up to 30% perforation level. For higher perforation of the normal plate, bubble is completely swept out. The peak turbulence value occurs around 0.7 to 0.8 times the reattachment length. The turbulence intensity values are highest for the case of solid normal plate (bleed air is absent) and are lowest for the case of 50% perforation of the normal plate (bleed air is maximum in the present study). From the analysis of data it is observed that $\sqrt{\overline{u^{{\prime}2}}}/(\sqrt{\overline{u^{{\prime}2}}})_{max}$, (the ratio of RMS velocity fluctuation to maximum RMS velocity fluctuation), is uniquely related with dimensionless distance y/Y', (the ratio of distance normal to splitter plate to the distance where RMS velocity fluctuation is half its maximum value) for all the perforated normal plates. It is interesting to note that for 50% perforation of the normal plate, the RMS pressure fluctuation in the flow field gets reduced to around 60% as compared to that for solid normal plate. Analysis of the results show that the ratio [$C^{\prime}_p$ max/$-C_{pb}(1-{\eta})$], where $C^{\prime}_p$ max is the maximum coefficient of fluctuating pressure, $C_{pb}$ is the coefficient of base pressure and ${\eta}$ is the perforation level (ratio of open to total area), for surface RMS pressure fluctuation levels seems to be constant and has value of about 0.22. Similar analysis show that the ratio $[C^{\prime}_p$ max/$-C_{pb}(1-{\eta})]$ for flow field RMS pressure fluctuation levels seems to be constant and has a value of about 0.32.

Chip-level NFP Calibration and Verification Using Improved Probe for NFS Standardization (NFS 표준을 위한 개선된 프로브를 이용한 칩 수준 NFP 측정값 교정 및 검증)

  • Lee, Pil-Soo;Wee, Jae-Kyung;Kim, Boo-Gyoun;Choi, Jai-Hoon;Yeo, Soon-Il
    • Journal of the Institute of Electronics Engineers of Korea SD
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    • v.49 no.6
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    • pp.25-34
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    • 2012
  • New calibration method for the near-field scanning (NFS) system is presented. The proposed calibration method consisted of a new near-field antenna (NFP) and newly devised patterns as circular patch patterns (CPPs) and meander patterns (MPs). The proposed patterns were used for improving spatial resolutions and simplifying a calibration procedure of the NFP compared to the conventional method defined in the IEC61967-3 and 6. Also, the effect of the length of NFPs on attenuation characteristics was investigated with length of 8mm and 30mm. For them, we designed and fabricated CPPs of diameter (D) = 20, 40, 60, and 100mm and MPs of various widths and spaces. We found the reverse relations between spatial resolutions and heights of measuring points by using simplified calibration procedure. The testing result shows that the spatial resolution of $120{\mu}m$ at height of $200{\mu}m$ was verified without complex correlation algorithms under 8GHz. For manufacturing cost all patterns and the NFP were realized with low-cost fabrication using PCB (FR-4) not by a conventional LTCC process. For verification of chip-level EMC from the results, near-field scanning system (NFSS) having step resolution of Sub-micron scale was produced and by using the proposed NFSS and proposed NFP measurement of chip shows accurately the shape of the resolution of $200{\mu}m$ patterns for securing a high level of chip-level EMC verification.