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Molecular Gene Cloning, Expression, and Characterization of Bovine Brain Glutamate Dehydrogenase

  • Kim, Dae-Won;Eum, Won-Sik;Jang, Sang-Ho;Yoon, Chang-Sik;Kim, Young-Hoon;Choi, Soo-Hyun;Choi, Hee-Soon;Kim, So-Young;Kwon, Hyeok-Yil;Kang, Jung-Hoon;Kwon, Oh-Shin;Cho, Sung-Woo;Park, Jin-Seu;Choi, Soo-Young
    • BMB Reports
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    • v.36 no.6
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    • pp.545-551
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    • 2003
  • A cDNA of bovine brain glutamate dehydrogenase (GDH) was isolated from a cDNA library by recombinant PCR. The isolated cDNA has an open-reading frame of 1677 nucleotides, which codes for 559 amino acids. The expression of the recombinant bovine brain GDH enzyme was achieved in E. coli. BL21 (DE3) by using the pET-15b expression vector containing a T7 promoter. The recombinant GDH protein was also purified and characterized. The amino acid sequence was found 90% homologous to the human GDH. The molecular mass of the expressed GDH enzyme was estimated as 50 kDa by SDS-PAGE and Western blot using monoclonal antibodies against bovine brain GDH. The kinetic parameters of the expressed recombinant GDH enzymes were quite similar to those of the purified bovine brain GDH. The $K_m$ and $V_{max}$ values for $NAD^+$ were 0.1 mM and $1.08\;{\mu}mol/min/mg$, respectively. The catalytic activities of the recombinant GDH enzymes were inhibited by ATP in a concentration-dependent manner over the range of 10 - $100\;{\mu}M$, whereas, ADP increased the enzyme activity up to 2.3-fold. These results indicate that the recombinant-expressed bovine brain GDH that is produced has biochemical properties that are very similar to those of the purified GDH enzyme.

Investigation on the reality of school mathematics based on the learner's competencies (학습자의 핵심역량에 기초한 수학교육 실태 탐색 - 뉴질랜드와 프랑스를 중심으로 -)

  • Choe, Seung-Hyun;Hwang, Hye-Jeang;Nam, Geum-Cheon
    • Journal of the Korean School Mathematics Society
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    • v.15 no.2
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    • pp.215-238
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    • 2012
  • One of the keyword in every nation's recent educational policy is key competencies. Considering national competitiveness originating from educational competitiveness, educational policy has been driven to identify key competencies and realize them through school education. Within this context some leading countries have developed competencies-based curriculum and discussed ways to relate key competencies and subject matter areas. However, there have been few researches on how to reflect or integrate key competencies into subject matter areas. Because of this reason, the ways to incorporate and integrate key competencies into three subject areas including mathematics were investigated. The recent trends of curriculum, teaching and learning, and assessment of domestic and foreign cases were explored by the subject of one Korean international middle school, one British foreign school in Seoul, one French foreign school in Seoul, and four middle schools in New Zealand. To establish competencies-based school education, there should be intimate connection system among curriculum, teaching and learning, assessment, and teacher education. Through analysis of domestic and foreign cases, some conclusions regarding how these aspects have changed with the emphasis of key competencies were drawn. In this paper, through classroom observations and teacher interviews, the reality of competencies-based mathematics teaching of New Zealand and France was investigated. As a result, summaries and recommendations related to ways to improve subject teaching and teacher education in light of key competencies were presented. In these recommendations, the ways to reconstruct subject-based curriculum, the content-specific teaching and learning, and educational assessment were included.

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Association of the thyroid hormone responsive spot 14 alpha gene with growth-related traits in Korean native chicken

  • Cahyadi, Muhammad;Park, Hee-Bok;Seo, Dong Won;Jin, Shil;Choi, Nuri;Heo, Kang Nyeong;Kang, Bo Seok;Jo, Cheorun;Lee, Jun Heon
    • Asian-Australasian Journal of Animal Sciences
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    • v.33 no.11
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    • pp.1755-1762
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    • 2020
  • Objective: Thyroid hormone responsive spot 14 alpha (THRSP) has been used to investigate the regulation of de novo lipogenesis because the variation of THRSP mRNA content in the tissue affects directly the ability of that tissue to synthetize lipids. Also, this gene responds to thyroid hormone stimulation and high level of carbohydrate feeding or insulin-injection. This study was carried out to investigate variations within THRSP and their effects on body and carcass weights in Korean native chicken (KNC). Methods: A total of 585 chickens which represent the five lines of KNC (Black, Gray-Brown, Red-Brown, White, and Yellow-Brown) were reared and body weight data were recorded every two weeks from hatch until 20 weeks of age. Polymerase chain reaction- restriction fragment length polymorphism, DNA chips for Agilent 2100 Bioanalyzer, and Fluidigm Genotyping Technology, were applied to genotype selected markers. A linear mixed-effect model was used to access association between these single nucleotide polymorphism (SNP) markers and growth-related traits. Results: A total of 30 polymorphisms were investigated in THRSP. Of these, nine SNPs for loci were selected to perform association analyses. Significant associations were detected between g.-49G>T SNP with body weight at 20 weeks of age (BW20), g.451T>C SNP with growth at 10 to 12 weeks of age (GR10-12), and g.1432A>C SNP with growth at 14 to 16 weeks trait (GR14-16) and body weight at 18 weeks of age (BW18). Moreover, diplotype of the THRSP gene significantly affected body weight at 12 weeks of age (BW12) and GR10-12 traits. Diplotype of ht1/ht2 was favorable for BW12 and GR10-12 traits. Conclusion: These results suggest that THRSP can be regarded as a candidate gene for growth traits in KNC.

Prevalence of Human Papillomavirus and Co-Existent Sexually Transmitted Infections among Female Sex Workers, Men having Sex with Men and Injectable Drug abusers from Eastern India

  • Ghosh, Ishita;Ghosh, Pramit;Bharti, Alok Chandra;Mandal, Ranajit;Biswas, Jaydip;Basu, Partha
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.3
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    • pp.799-802
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    • 2012
  • Background: Human papillomavirus (HPV) is a very common sexually transmitted disease affecting both men and women and is responsible for different ano-genital cancers in either sex. Co-existing sexually transmitted infections (STI) including HIV have been considered as important co-factors for carcinogenesis induced by HPV. The purpose of this study was to determine the prevalence of any HPV, HPV 16 and HPV 18 and also concomitant STIs among female sex workers (FSW), men having sex with men (MSM) and injectable drug users (IDU). Material and Method: This cross-sectional study was conducted among 45 FSWs, 26 MSMs and 58 IDUs who attended the STI or de-addiction clinics. Genital scrape samples collected from glans penis and coronal sulcus in males and cervical squamo-columnar junction in females were tested for HPV DNA by PCR using HPV L1 consensus primer. Type specific PCR to detect HPV 16 and 18 was done on the samples positive on consensus PCR. All participants were tested for associated STIs including HIV and hepatitis B and cervical cytology was done on all females. Results: Among the FSWs, HPV was detected in 73.3% and HPV 16 and 18 was detected in 25.7%. Though the HPV prevalence was similarly high among MSMs (69.2%) and IDUs (72.4%), the prevalence of HPV 16 and 18 was much lower in these groups compared to the FSWs. Prevalence of cervico-vaginal infection with Trichomonas vaginalis and syphilis was significantly higher in the HPV positive women compared to the HPV negative women. There was no statistically significant difference in the prevalence of other STIs among HPV positive and negative women and men. Conclusion: HPV infection is highly prevalent among FSW, MSM and IDUs. Trichomonas vaginalis infection is more frequent in HPV positive women.

A Study on the Characteristics and Identity of Public Art in Historical and Cultural Areas - Focused on Taoxichuan in Jingdezhen - (역사문화지구의 퍼블릭 아트에서 나타나는 표현 특성과 아이덴티티 연구 - 징더전 시 도계천을 중심으로 -)

  • Li, JianHua;Yoon, Ji-Young
    • The Journal of the Korea Contents Association
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    • v.20 no.2
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    • pp.82-95
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    • 2020
  • This study takes the identity and characteristics of public art in historical and cultural regions as the research object. Through the theoretical investigation and research on the public art of historical and cultural regions, six evaluation factors that shape spatial identity and characteristics were drawn: Environmental characteristics, public characteristics; Symbolic characteristics; Artistic characteristics;.Historical characteristics; Interactivity. The identity and characteristics of Taoxichuan public art in Jingdezhen was analyzed and evaluated. The results show that the " public art " of seven large chimneys is consistent with the historical structure of the industry. Among them, environmental, public characteristics, symbolic, artistic and historical characteristics of the Industria park have a high degree of preservation value and can be recognized as the historical landmark of the park. In the new public art, the artistic quality of public art, the environment, and the public characteristic of public art, are also at a moderate level, but it is comparatively weak. Public art recreated as an industrial heritage has formed an identity that can communicate spiritual sympathy to the public by forming a time-space communication between the public with historical relics and ceramic elements, and the new public art for local rehabilitation has given new artistic flavor to the city, but the new identity has not been shown.

A Study on Article 78 CISG: Interest on Sums in Arrears (CISG 제78조(연체이자(延滯利子) 청구권(請求權))에 대한 고찰(考察))

  • Kim, Tae-Gyeong
    • THE INTERNATIONAL COMMERCE & LAW REVIEW
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    • v.31
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    • pp.3-25
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    • 2006
  • This study focus on interest for arrears and filling of the gaps left in Article 78 of CISG. In the case of CISG, Article 78 provides for interest any time a payment under a contract is untimely, but does not specify a particular rate of interest or a method to determine such a rate. This issue did not cause any uncertainty under ULIS, the CISG's antecedent, since Article 83 of ULIS provided for 1%p above the official discount rate in the creditor's country. Lacking any CISG general principle as well as any indication by the very same CISG, one can only conclude that the matter must be deferred to the domestic rule of private international law. Actually, resorting to private international law is not only admissible, but expressly required by Article 7(2). In the interpretation and filling of the gaps left in Article 78, there is a considerable difference of opinion especially amongst commentators on whether the gap is a lacuna praeter legem, i.e., one being governed by, but not expressly settled in the CISG, or whether it is an issue falling outside the scope of application of the CISG, i.e. a lacuna intra legem. The protagonists of the former view lay emphasis on the overall objective of the CISG, namely to create a uniform law, whereas the supporters of the latter view refer to the legislative history of Article 78 as the dominant principle in interpreting Article 78. Some authors believe that the issue of determining the rate of interest is not dealt with by CISG and it is, therefore, governed by the applicable domestic law, which is the subsidiary law applicable to the sales contract, since "no special connecting points seem to have developed for the entitlement to interest." In the light of the relevant case law, it seems correct to conclude that the interest rate is not determined by CISG and that courts normally determine it according to their own rules of private international law. While CISG Article 78 expressly does not deal with this issue, PICC Article 7.4.9 and PECL Article 9.508, on the other hand, set forth a precise method for computing interest. Although a method like the one set by PICC may be useful and may encourage uniformity, it still cannot be used under the CISG. The PICC or PECL formula may, however, be a very good starting point in a de jure condendum analysis when a new Article 78 will be drafted, if an interest rate method will ever be embodied in the text of an international convention.

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Translational Study for "Treatise on Bundon Disease in Chapter 8 of the Essence of the Synopsis of the Golden Chamber" ("금궤요략심전.분돈기병맥증치제팔(奔豚氣病脈證治第八)"에 대한 번역연구)

  • Li, Xian-Lan;Lee, Yong-Bum
    • Journal of Korean Medical classics
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    • v.21 no.2
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    • pp.21-28
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    • 2008
  • After translating the contents of "Treatise on Bundon Disease in Chapter 8 of the Essence of the Synopsis of the Golden Chamber" and it was compared with the contents of "The Lecture of Synopsis of Prescriptions of the Golden Chamber" and speculated to review the characteristics of the comments of Ui(尤怡) and the following conclusions were made. As the causes of bundon(奔豚) disease, which means gas rushing like a running piggy from the lower abdomen to the throat, they could be categorized into the cases belongs to miscellaneous diseases (雜病) and into the cases of suffering from cold diseases(傷寒). If it was diagnosed as miscellaneous disease, terror and fear(驚恐) could be regarded as its cause and if it was diagnosed as cold disease, insufficiency of heart Gi(心氣) due to the excessive sweating and the invasion of exterior coldness could be regarded as its cause. As the pathogenesis of this disease, the pathogenic factors of liver and kidney could be found. Since the liver and Soyang meridian(少陽經) are subjected to the exterior and interior relation in case of the pathogenic factor of liver, alternate attacks of chills and fever(寒熱往來) could be characteristic for the disease. If this disease was caused by the pathogenic factor of kidney, there are two cases. One is the palpitation under the umbilicus(欲作奔豚: 臍下動悸) developed by the lack of heart Gi(心氣) and the other is gas rushing like a running piggy from the lower abdomen to the throat which could be developed by the exterior cold disease which invade into the interior and reach into the kidney when the heart Gi(心氣) was insufficient. In its treatment, bundon disease caused by the pathogenic factor of liver could be treated with Bundontang(奔豚湯), and the understanding of Ui(尤怡) on Pinellia ternata Breitenbach(半夏) having the effect of perspiration in the prescription is quite unique concept. In case of bundon disease caused by the pathogenic factor of kidney, treatment is focused to treat exterior coldness by applying moxibustion(灸) and Gyejigagyetang(桂技加桂湯), and in case of the palpitation under the umbilicus(欲作奔豚: 臍下動悸), its pathogenic factor of kidney is controlled with Cinnamomum louerii Nees(桂技), Polia cocos Walf and the spleen Gi(脾氣) is cared by using Glycyrrhiza uralensis Fischer et. De Candolle(甘草) and Zizyphus jujuba Mill(大棗). Especially, Ui(尤怡) emphasized Cinnamomum louerii Nees(桂技) as the medication to treat the pathogenic factor of kidney.

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Effects of L-carnitine, Selenium-enriched Yeast, Jujube Fruit and Hwangto (Red Clay) Supplementation on Performance and Carcass Measurements of Finishing Pigs

  • Han, Yung-Keun;Thacker, P.A.
    • Asian-Australasian Journal of Animal Sciences
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    • v.19 no.2
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    • pp.217-223
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    • 2006
  • Fifty castrated crossbred ($Landrace{\times}Yorkshire$) pigs, weighing an average of $60.6{\pm}3.1kg$ were allotted to one of five treatments in a randomized block design to examine the effects of dietary inclusion of 0.1% L-carnitine (50 ppm carnitine), 0.1% selenium-enriched yeast (0.3 ppm selenium), 0.1% Jujube fruit or 0.1% Hwangto (Red clay) on pig performance and carcass quality. All diets were based on corn, wheat, soybean meal and wheat bran and were formulated to supply 13.8 MJ DE/kg. Dietary supplementation did not influence daily gain (p = 0.57), feed intake (p = 0.52), or feed conversion (p = 0.32). Digestibility of dry matter (p = 0.60), organic matter (p = 0.74), crude protein (p = 0.76), crude fibre (p = 0.70) and energy (p = 0.75) were also unaffected by inclusion of any of the additives. Tissue samples taken from the longissimus muscle showed that the levels of carnitine (p = 0.0001) and selenium (p = 0.0001) were significantly higher with dietary inclusion of carnitine or selenium-enriched yeast. Dietary treatment did not affect dressing percentage (p = 0.33), carcass lean yield (p = 0.99) or first, $10^{th}$ and last rib midline backfat depth (p = 0.45, 0.82 and 0.47, respectively). Dietary treatment also did not affect the percentages of tenderloin (p = 0.37), bacon (p = 0.36), fat and bone (p = 0.56), picnic shoulder (p = 0.25), skirt (p = 0.80), fresh ham (p = 0.31) or ribs (p = 0.79). However, pigs fed the diet containing Jujube fruit had a higher percentage of Boston butt than pigs fed the carnitine or selenium supplemented diets (p = 0.01). Pigs fed added Hwangto had a higher (p = 0.04) percentage of loin compared with pigs fed supplementary selenium or Jujube fruit. Loin muscle from pigs fed carnitine had a significantly lower Hunter colour value for L (whiteness, p = 0.004) and a higher value for $a^*$ (redness; p = 0.069). The overall results indicate that supplementation with L-carnitine and selenium-enriched yeast can produce pork containing higher levels of carnitine and selenium, which could provide health benefits for consumers of pork without detrimental effects on pig performance.

Performance and Metabolism of Calves Fed Starter Feed Containing Sugarcane Molasses or Glucose Syrup as a Replacement for Corn

  • Oltramari, C.E.;Napoles, G.G.O.;De Paula, M.R.;Silva, J.T.;Gallo, M.P.C.;Pasetti, M.H.O.;Bittar, C.M.M.
    • Asian-Australasian Journal of Animal Sciences
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    • v.29 no.7
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    • pp.971-978
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    • 2016
  • The aim of this study was to evaluate the effect of replacing corn grain for sugar cane molasses (MO) or glucose syrup (GS) in the starter concentrate on performance and metabolism of dairy calves. Thirty-six individually housed Holstein male calves were blocked according to weight and date of birth and assigned to one of the starter feed treatments, during an 8 week study: i) starter containing 65% corn with no MO or GS (0MO); ii) starter containing 60% corn and 5% MO (5MO); iii) starter containing 55% corn and 10% MO (10MO); and iv) starter containing 60% corn and 5% GS (5GS). Animals received 4 L of milk replacer daily (20 crude protein, 16 ether extract, 12.5% solids), divided in two meals (0700 and 1700 h). Starter and water were provided ad libitum. Starter intake and fecal score were monitored daily until animals were eight weeks old. Body weight and measurements (withers height, hip width and heart girth) were measured weekly before the morning feeding. From the second week of age, blood samples were collected weekly, 2 h after the morning feeding, for glucose, ${\beta}$-hydroxybutyrate and lactate determination. Ruminal fluid was collected at 4, 6, and 8 weeks of age using an oro-ruminal probe and a suction pump for determination of pH and short-chain fatty acids (SCFA). At the end of the eighth week, animals were harvested to evaluate development of the proximal digestive tract. The composition of the starter did not affect (p>0.05) concentrate intake, weight gain, fecal score, blood parameters, and rumen development. However, treatment 5MO showed higher (p<0.05) total concentration of SCFAs, acetate and propionate than 0MO, and these treatments did not differ from 10MO and 5GS (p>0.05). Thus, it can be concluded that the replacement of corn by 5% or 10% sugar cane molasses or 5% GS on starter concentrate did not impact performance, however it has some positive effects on rumen fermentation which may be beneficial for calves with a developing rumen.

Physicochemical characteristics and fatty acid profiles of muscle tissues from Hanwoo steers fed a total mixed ration supplied with medicinal plant by-products

  • Lee, Shin Ja;Kim, Do Hyung;Yang, Han Sul;Nam, Ki Chang;Ahn, Seung Kyu;Park, Sung Kwon;Choi, Chang Weon;Lee, Sung Sill
    • Asian-Australasian Journal of Animal Sciences
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    • v.30 no.10
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    • pp.1388-1395
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    • 2017
  • Objective: Using medicinal plant by-products (MPBP) as feed additives may be an eco-friendly option as substitutes for feedstuffs and may assist in reducing the improper disposal of MPBP. Therefore, this study was conducted to evaluate the influences of MPBP on the meat quality of Hanwoo steers fed a total mixed ration (TMR). Methods: Twenty seven steers (body weight = $573{\pm}57kg$) were randomly divided into three treatments with a control group and two tested groups as follows: control, 1,000 g/kg TMR; treatment 1 (MPBP30), 970 g/kg TMR and 30 g/kg MPBP; treatment 2 (MPBP50), 950 g/kg TMR and 50 g/kg MPBP. Results: Average daily gain, feed conversion ratio and the Commission Internationale de l'Eclairage $L^*$ of muscle were improved (p<0.05, respectively) by MPBP30. Stearic acid ($C_{18:0}$) was decreased (linear effect, p = 0.012), while oleic acid ($C_{18:1}$) was increased (linear effect, p = 0.055) by MPBP level. Saturated fatty acid (SFA) and polyunsaturated fatty acid (PUFA) were decreased for MPBP50 while unsaturated fatty acid (USFA) and monounsaturated fatty acid (MUFA) were increased for MPBP 50. USFA and SFA ratio was increased for MPBP50 as well. Conclusion: These results indicated that MPBP supplementation in Hanwoo steers fed a TMR increased feed efficiency and meat color (lightness) with altering fatty acid proportions. Therefore, MPBP may be successfully used in ruminant feeding.