• Journal of Plant Biotechnology
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• 제49권4호
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• pp.331-338
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• 2022

Sunflower (Helianthus annuus L.) protoplasts were isolated from seven-day-old etiolated hypocotyls of 10 A line and four-week-old fully expanded young leaves of PI 441983 line in vitro seedlings using an enzymatic method. Purified protoplasts were collected by filtration and floatation in sucrose solution. Semi-solid protoplast culture was performed using the L4 regeneration protocol with various culture media and plating densities to achieve the highest efficiencies for protoplast culture of hypocotyl and mesophyll protoplasts of 10 A and PI 441983 lines, respectively. The concentrations in liquid L'4M medium and different plating densities were evaluated in two types of cytokinins, the adenine-type 6-benzyladenine (BA) and the phenylurea-type thidiazuron (TDZ). The highest colony formation was achieved in both sunflower lines when 0.5 mgL^-1 BA and 0.5 mgL^-1 TDZ were applied with a high plating density (3 × 10⁵ protoplasts mL^-1). These conditions led to 38.45% and 39.40% colony formation for hypocotyl protoplasts of the 10 A line and mesophyll protoplasts of the PI 441983 line, respectively. Moreover, many hypocotyl protoplast-derived colonies developed into micro-calli. In addition, superior development of both sunflower protoplasts was observed with all plating densities when BA was used in combination with TDZ. This finding will be applicable to future sunflower hybrid production via somatic hybridization.

• 멤브레인
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• 제33권6호
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• pp.269-278
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• 2023

효소 고정화 막 생물반응기(EMBRs)는 폐수 내의 염료를 처리하는 새로운 방법입니다. 이 분야는 효소의 효능과 환경에 대한 높은 저항성 때문에 많은 양의 연구가 진행되었습니다. 효소 자체와 해당 효소의 구조를 모두 포함하는 다양한 방법이 EMBR에 접근할 수 있습니다. 생물반응기 자체는 염료 제거의 필요에 맞게 변형될 수 있습니다. 효소적 생물반응기부터 산화 그래핀 또는 탄소 나노튜브와 같은 나노구조를 사용하는 것까지 다양합니다. 또한 TiO₂와 같은 나노입자는 EMBR을 더욱 향상시키기 위해 사용될 수 있습니다. 폴리머 기반의 막 지지 구조는 또한 효능 증가의 문제에 접근하는 다양한 방법을 포함합니다. 본 바와 같이, 지난 수십 년 동안 EMBR을 사용하는 이 문제에 대한 다양한 접근법이 수행되었습니다. 이 검토는 방법론을 요약하고 EMBR에 대한 다양한 개선 사항을 설명하는 것을 목표로 합니다.

• Animal Bioscience
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• 제36권11호
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• pp.1693-1699
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• 2023

Objective: Cows that are nursing get around 80% of their glucose from liver gluconeogenesis. Propionate, a significant precursor of liver gluconeogenesis, can regulate the key genes involved in hepatic gluconeogenesis expression, but its precise effects on the activity of enzymes have not yet been fully elucidated. Therefore, the aim of this study was to investigate the effects of propionate on the activity, gene expression, and protein abundance of the key enzymes involved in the gluconeogenesis of dairy cow hepatocytes. Methods: The hepatocytes were cultured and treated with various concentrations of sodium propionate (0, 1.25, 2.50, 3.75, and 5.00 mM) for 12 h. Glucose content in the culture media was determined by an enzymatic coloring method. The activities of gluconeogenesis related enzymes were determined by enzyme linked immunosorbent assay kits, and the levels of gene expression and protein abundance of the enzymes were detected by real-time quantitative polymerase chain reaction and Western blot, respectively. Results: Propionate supplementation considerably increased the amount of glucose in the culture medium compared to the control (p<0.05); while there was no discernible difference among the various treatment concentrations (p>0.05). The activities of cytoplasmic phosphoenolpyruvate carboxylase (PEPCK1), mitochondrial phosphoenolpyruvate carboxylase (PEPCK2), pyruvate carboxylase (PC), and glucose-6-phosphatase (G6PC) were increased with the addition of 2.50 and 3.75 mM propionate; the gene expressions and protein abundances of PEPCK1, PEPCK2, PC, and G6PC were increased by 3.75 mM propionate addition. Conclusion: Propionate encouraged glucose synthesis in bovine hepatocytes, and 3.75 mM propionate directly increased the activities, gene expressions and protein abundances of PC, PEPCK1, PEPCK2, and G6PC in bovine hepatocytes, providing a theoretical basis of propionate-regulating gluconeogenesis in bovine hepatocytes.

• 한국식품저장유통학회지
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• 제30권6호
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• pp.929-943
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• 2023

This study analyzed the variation in vitamin K content in conger eel (Conger myriaster) caught off Tongyeong, Gyeongsangnam-do, Korea, focusing on the influence of size (large and small) and harvest period (monthly throughout 2021). We applied enzymatic extraction and HPLC-fluorescence methods for the analysis of vitamin K₁ (phylloquinone) and K₂ (menaquinone). The vitamin K content in conger eel varied significantly with size and harvest season (p<0.05). In large-sized samples, the phylloquinone content peaked in July (0.80±0.09 ㎍/100 g), while the highest menaquinone content was in May (0.79±0.11 ㎍/100 g). Conversely, in small-sized conger eels, the highest phylloquinone was found in December (1.94±0.15 ㎍/100 g), and the peak menaquinone level was in January (0.66±0.02 ㎍/100 g). The fat content was highest in July for large samples and in January for smaller ones. There was a positive correlation between fat and total vitamin K contents in conger eel (r=0.631, 0.667). Method validation and quality control measures ensured data reliability for vitamin K₁ and K₂ analyses. This study provides reliable information on the size and seasonal variations of vitamin K in conger eels, a staple in the Korean diet. This information is valuable for inclusion in Korea's national food nutrition database and for formulating future national health and nutrition policies.

• Journal of Periodontal and Implant Science
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• 제53권5호
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• pp.321-335
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• 2023

Purpose: The aim of this study was to investigate the efficacy of photo-crosslinked gelatin methacryloyl (GelMa) hydrogel containing calcium phosphate nanoparticles (CNp) when applying different fabrication methods for bone regeneration. Methods: Four circular defects were created in the calvaria of 10 rabbits. Each defect was randomly allocated to the following study groups: 1) the sham control group, 2) the GelMa group (defect filled with crosslinked GelMa hydrogel), 3) the CNp-GelMa group (GelMa hydrogel crosslinked with nanoparticles), and 4) the CNp+GelMa group (crosslinked GelMa loaded with nanoparticles). At 2, 4, and 8 weeks, samples were harvested, and histological and micro-computed tomography analyses were performed. Results: Histomorphometric analysis showed that the CNp-GelMa and CNp+GelMa groups at 2 weeks had significantly greater total augmented areas than the control group (P<0.05). The greatest new bone area was observed in the CNp-GelMa group, but without statistical significance (P>0.05). Crosslinked GelMa hydrogel with nanoparticles exhibited good biocompatibility with a minimal inflammatory reaction. Conclusions: There was no difference in the efficacy of bone regeneration according to the synthesized method of photo-crosslinked GelMa hydrogel with nanoparticles. However, these materials could remain within a bone defect up to 2 weeks and showed good biocompatibility with little inflammatory response. Further improvement in mechanical properties and resistance to enzymatic degradation would be needed for the clinical application.

• 한국근적외분광분석학회:학술대회논문집
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• 한국근적외분광분석학회 2001년도 NIR-2001
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• pp.1265-1265
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• 2001

The organic materials included in excreta of livestock are important resources for organic manure and for improving soil quality, although there is still far from effective using. One reason for this is still unclearly standard of quality for evaluation of manure made from excreta of livestock. Therefore, the objective of this study is to develop rapid and accurate analytical method for analyzing organic compositions of manure made from excreta of livestock, and to establish quality evaluation method based on the compositions predicted by near infrared reflectance spectroscopy (NIRS). Sixteen samples of manure, each eight samples prepared from two treatments, were used in this study. The manure samples were prepared by mixing 560 kg feces of swine,60 kg sawdust with moisture content was adjusted to be 65%. The mixture was then keep under two kinds of shelter, black and clear sheets, as a treatment on the effect of sunlight. Samples were taken in every week (form week-0 to 7) during the process of manure making. Samples were analyzed to determine neutral detergent fiber (NDF), acid detergent fiber (ADF) and acid detergent lignin (ADL) by detergent methods, and organic cell wall (OCW) and fibrous content of low digestibility in OCW (Ob) by enzymatic methods. Biological oxygen demand (BOD) was analyzed by coulometric respirometer method. These compositions were carbohydrateds and lignin that were hardly digested. Spectra of samples were scanned by NIR instrument model 6500 (Pacific Scientific) and read over the range of wavelength between 400 and 2500nm. Calibration equations were developed using eight manure samples collected from black sheet shelter, while prediction was conducted to the other eight samples from clear sheet shelter. Accuracy of NTRS prediction was evaluated by correlation coefficients (r), standard error of prediction (SEP) and ration of standard deviation of reference data in prediction sample set to SEP (RPD). The r, SEP and RPD value of forage were 0.99, 0.69 and 7.6 for ADL, 0.96, 1.03 and 4.1 for NDF, 0.98, 0.60 and 4.9 for ADF, 0.92, 1.24 and 2.6 for Ob, and 0.91, 1.02 and 7.3 for BOD, respectively. The results indicated that NIRS could be used to measure the organic composition of forage used in manure samples.

• Journal of Ginseng Research
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• 제42권2호
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• pp.149-157
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• 2018

Background: Panax notoginseng leaves (PNL) exhibit extensive activities, but few analytical methods have been established to exclusively determine the dammarane triterpene saponins in PNL. Methods: Ultra-performance liquid chromatography coupled with time-of-flight mass spectrometry (UPLC/Q-TOF MS) and HPLC-UV methods were developed for the qualitative and quantitative analysis of ginsenosides in PNL, respectively. Results: Extraction conditions, including solvents and extraction methods, were optimized, which showed that ginsenosides Rc and Rb3, the main components of PNL, are transformed to notoginsenosides Fe and Fd, respectively, in the presence of water, by removing a glucose residue from position C-3 via possible enzymatic hydrolysis. A total of 57 saponins were identified in the methanolic extract of PNL by UPLC/Q-TOF MS. Among them, 19 components were unambiguously characterized by their reference substances. Additionally, seven saponins of PNL-ginsenosides Rb1, Rc, Rb2, and Rb3, and notoginsenosides Fc, Fe, and Fd-were quantified using the HPLC-UV method after extraction with methanol. The separation of analytes, particularly the separation of notoginsenoside Fc and ginsenoside Rc, was achieved on a Zorbax ODS C8 column at a temperature of $35^{\circ}C$. This developed HPLC-UV method provides an adequate linearity ($r^2$ > 0.999), repeatability (relative standard deviation, RSD < 2.98%), and inter- and intraday variations (RSD < 4.40%) with recovery (98.7-106.1%) of seven saponins concerned. This validated method was also conducted to determine seven components in 10 batches of PNL. Conclusion: These findings are beneficial to the quality control of PNL and its relevant products.

• 한국산림과학회지
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• 제39권1호
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• pp.57-63
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• 1978

Trichoderma viride SANK호(號) cellulase에 의(依)한 목재(木材) 가수분해(加水分解)에 관(關)한 연구(硏究)로서 재료(材料)는 산오리나무재(材)를 사용(使用)하였다. 그리고 당생성(糖生成)을 위한 cellulase의 최적(最適) 반응조건(反應條件)을 조사(調査)하였다. Trichoderma viride 조효소액(粗酵素液)은 진탕배양법(振盪培養法)에 의(依)하여 생산(生產)하였고, 이것을 염석효소액(鹽析酵素液)을 조제(調製)하여 사용(使用)하였다. 기질(基質)은 산오리나무재(材) 톱밥을 과초산법(過醋酸法)에 의(依)하여 탈(脫)리그닌 한것을 사용(使用)하였다. 환원당(還元糖) 정량(定量)은 DNS법(法)에 의(依)하였다. 결과(結果)는 다음과 같다. 1. 최적(最適) pH는 5.0이며, pH 안정성(安定性)의 범위(範圍)는 4.0~6.0으로 나타났다. 2. 반응온도(反應溫度)는 $40^{\circ}C$가 최적(最適)이였으며, 온도(溫度)의 안정성(安定性)은 $40^{\circ}C{\sim}50^{\circ}C$ 범위(範圍)로 나타났다. 3. 효소농도(酵素濃度)는 높을수록 환원당(還元糖) 생성율(生成率)은 증가(增加)되었다. 4. 기질농도(基質濃度)는 높을수록 단위(單位) 기질(基質)에 대(對)한 환원당(還元糖) 생성율(生成率)은 감소(減少)되었다. 5. 당류중(糖類中) pructose가 가장 강(强)한 조해제(阻害劑)로 나타났으며, glucose의 조해작용(阻害作用)은 약(弱)하게 나타났다.

• Tuberculosis and Respiratory Diseases
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• 제42권6호
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• pp.838-845
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• 1995

연구배경: 현재 국내에서 ADA의 활성치의 측정은 adenosine 기질에서 ADA에 의한 탈아미노작용시 발생하는 암모니아를 Berthelot's 반응을 이용하여 측정하는 Giusti 등에 의한 비색법이 많이 이용되고 있다. 그러나 이 방법은 자동화가 어려워 쉽게 사용하기 어려우며 기기내의 내인성 암모니아 등에 의한 오염의 가능성이 있다. 1993년 Oosthuizen 등은 nucleoside phosphorylase(NP)와 Xanthine oxidase(XOD)을 이용하여 ADA 활성치 측정을 자동화 하였다. 저자들이 측정한 ADA 활성치와 기존의 ADA 활성치 측정결과에 대한 보고를 비교함으로써 Oosthuizen 등에 의한 ADA 활성치 측정의 자동화의 융통성을 알아보고자 하였다. 방법: 1994년 5월부터 1995년 7월까지 연세대학교 원주의과대학부속 원주기독병원에 흉막액으로 입원하여 그 원인이 확진된 162명의 환자를 각각의 원인 질환에 따라서 5개 군(I: 결핵성 흉막액, II: 악성 흉막액, III: 폐렴성 흉막액, V: 여출성 흉막액)으로 나누었으며 Oosthuizen 등에 의한 ADA 활성치 측정법을 Hitachi 747 자동화학분석기에 적용하여 각 군에서 흉막액의 ADA 활성치와 및 흉막액과 혈청에서 ADA 활성치의 비를 측정하였다. 결과: 1) 결핵성 흉막액의 ADA 활성치는 $52.53{\pm}16.43\;U/L$로서 나머지 군에 비하여 통계학적으로 의의있게 높았으며(II, IV, V 군은 p값이 0.001 미만, III 군은 p값이 0.05 미만) 흉막액에서 ADA의 활성치를 30 U/L로 기준하였을 때 결핵성 흉막액과 악성 흉막액의 감별은 민감도(sensitivity) 96%, 특이도(specificity) 93%로 결핵성 흉막염을 진단할 수 있었으며 이는 기존의 보고와 차이가 없었다. 2) 흉막액과 혈청 ADA 치의 비는 결핵성 흉막액에서 $2.29{\pm}0.96$으로 농흉을 제외한 나머지 군에 의해서 통계학적으로 의의있게 높았다(p<0.001). 흉막액과 혈청 ADA 활성치 비를 1.5로 기준하였을 때 결핵성 흉막액과 악성 흉막액의 감별은 민감도 80%, 특이도 88%로 결핵성 흉막액을 진단할 수 있었으며, 이는 기존의 연구 결과와 차이가 없었다. 3) Oosthuizen 동에 의한 ADA 활성치 측정법을 Hitachi 747 자동화학분석기에 적용한 새로운 방법은 r값이 0.971로 Giuisti 등에 의한 기존의 방법과 높은 상관관계를 보였다. 결론: ADA 활성치 측정에 Oosthuizen 등에 제안한 새로운 방법은 현재 사용되는 Giusti 등의 비색법에 의한 ADA 활성치 측정과 차이가 없었으며, 일반 검사실에서도 쉽게 사용될 수 있는 장점이 있었다.

• 한국응용곤충학회지
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• 제16권4호
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• pp.199-208
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• 1977

수종의 식물병원균(흰비단병균, 균핵병균, 좀검은균 핵병균)이 생산하는 가수분해산소(Cellulase Cx, Invertase, $\beta-amylase$, Xylanase, PMG, PG, Phosphatase, Protease)를 균계체내의 효소 및 배양여액내의 효소와 균핵내의 효소(좀검은균핵병균은 제외)로 나누어 그의 생산량과 pH에 따른 활성의 변화를 검토한 결과를 요약하면 다음과 같다. 1. 배양 10일후의 Cx활성은 균핵병균이 다른 균에 비하여 활성이 높았고 흰비단병균과 균핵병균은 산성측(pH 3.0 부근)에서, 좀검은균핵병균은 중성측(pH6.0 부근)에서 활성이 높았다. 2. 흰비단병균의 Invertase는 다른균에 비하여 약 20배정도 높은 활성을 보였고 3균주 모드 배양여액과 균계체간에 효소활성의 차가 인정되지 않았다. 3. Xylanase의 활성은 3균주 모두 균계체, 균핵 및 배양여액에 따라 또 pH에 따라 아주 다양한 변화를 나타냈고 균핵내에서 활성이 높았다. 4. $\beta-amylase$의 활성은 공시균중 좀검은균핵병균의 균계체, 배양여액이 가장 높았다. (약 12.0ug/min) 흰비단병균과 균핵병균ㅇ서는 균계체나 균핵에서 보다 배양여액에서 높았는데 활성최적pH는 균계체, 균핵 모두 pH 6.2였으나 배양여액에서는 흰비단 병균과 균핵병균이 pH3.0이었는데 반해 좀검은균핵병균은 pH6.2였다. 5. PMG의 활성은 공시균 모두 배양여액에서 높았고 균계체에서는 균핵병균과 좀검은균핵병균이 높았으며 활성최적 pH는 균에 따라 또는 측정부분에 따라 다양하게 나타났다. 6. PG의 활성은 흰비단 병균과 균핵병균의 균계체에서 각각 9,1ug/min. 9.5ug/min으로서 가장 높았고 활성최적 pH는 흰비단병균이 pH4.5부근 균핵병균이 pH.3.0부근이었다. 7. 흰비단병균과 균핵병균의 Phosphatase는 산성측(최적 pH3.5)에서 활성이 높았고 좀검은 균핵 병균은 산성, 중성, 알카리측에서 각각 Peak가 나타났으나 최적 pH는 9.5였다. 8. 공시균주 모두 Protease는 pH 10.0에서 최고활성을 나타냈고 특히 좀검은균핵 병균의 배양여액내 효소활성이 높았다.