• Title/Summary/Keyword: Molecular approach

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Biosynthesis of Medium-chain-length Polyhydroxylalkanoate Using Extracted Oil from Spent Coffee Grounds (커피 찌꺼기 오일을 이용한 medium-chain-length Polyhydroxylalkanoate생합성)

  • Hui-Yeon Kwon;Jong-Sik Kim;Chung-Wook Chung
    • Journal of Life Science
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    • v.34 no.10
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    • pp.682-690
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    • 2024
  • This study aimed to produce medium-chain-length polyhydroxyalkanoates (mcl-PHA) using oil extracted from spent coffee grounds (SCG) as a carbon source. The highest oil yields, 14.40%(w/w) and 14.49%(w/w), were achieved with an extraction time of 2 hr and an SCG/n-hexane ratio of 15:1. The fatty acid composition of the extracted SCG oil (EOC) primarily consisted of linoleic acid (45.08%), palmitic acid (35.56%), oleic acid (8.62%), and stearic acid (7.62%). Pseudomonas sp. HY61, isolated from soil, was used to biosynthesize mcl-PHA composed of 3-hydroxyhexanoate, 3-hydroxyoctanoate, 3-hydroxydecanoate, and 3-hydroxydodecanoate using EOC. Under optimal flask conditions, the highest cell growth and mcl-PHA accumulation were observed with 5 g/l EOC, 72 hr, with NH4Cl as the nitrogen source, at pH 7, 25℃, and a C/N ratio of 15:1. Batch fermentation resulted in a dry cell weight (DCW) of 0.92 g/l and mcl-PHA of 0.17 g/l after 72 hr. In contrast, fed-batch fermentation, with additional supplementation of EOC and NH4Cl at 18, 30, and 42 hr, increased the DCW to 2.85 g/l and mcl-PHA accumulation to 0.67 g/l, approximately three times higher than batch fermentation. Gas chromatography confirmed that the mcl-PHA structure was consistent across both fermentation modes. The synthesized mcl-PHA had higher molecular weight and thermal decomposition temperatures compared to other mcl-PHA, suggesting its potential applications in packaging and biomedical fields. This study offers an economical approach for mcl-PHA production using waste EOC and Pseudomonas sp. HY61.

Expression of Bcl-2 Family in 4-Nitroquinoline 1-Oxide-Induced Tongue Carcinogenesis of the Rat (백서 혀에서의 4-nitroquinoline 1-oxide 유도 발암과정에서 Bcl-2 계 유전자의 발현)

  • Choi, Jae-Wook;Chung, Sung-Su;Lee, Geum-Sug;Kim, Byung-Gook;Kim, Jae-Hyeong;Kook, Eun-Byul;Jang, Mi-Sun;Ko, Mi-Kyeong;Jung, Kwon;Choi, Hong-Ran;Kim, Ok-Joon
    • Journal of Oral Medicine and Pain
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    • v.30 no.3
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    • pp.301-317
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    • 2005
  • The number of patients with tongue carcinoma is increasing rapidly among young individuals in many parts of the world. Oral carcinoma progresses from hyperplastic lesion through dysplasia to invasive carcinoma and the concept of "field cancerization" with molecular alteration has been suggested for oral cavity carcinogenesis. Significant improvement in treatment and prognosis will depend on more detailed understanding of the multi-step process leading to cancer development. To induce tongue carcinoma in rat by 4-NQO, each drinking water was made to 10 ppm, 25 ppm, 50 ppm and control (only D.W. without 4-NQO). Specimens were classified into 4 groups such as control, I (mild & moderate dysplasia), II (severe dysplasia and carcinoma in situ), III (carcinoma). The mRNA expressions of Bcl-2 family were evaluated by RT-PCR technique. For anti-apoptotic Bcl-2 family, mRNA expression of Bcl-w was down-regulated in all stages of tongue carcinogenesis model. However, mRNA expression of Bcl-2 was up-regulated. For pro-apoptotic Bcl-2 family, all members were down-regulated in all stages of tongue carcinogenesis model except for Bad mRNA in group III. In terms of BH3 only protein, mRNA expressions of Bok and Mcl-1 were down regulated in all stages of specimen, but Bmf in group II and BBC3 in group III were up-regulated. Our current findings demonstrated the involvements of mRNA expression of Bcl-2 family in multi-step tongue carcinogensis. This highlights the necessity for continued efforts to discover suitable biomakers (Bcl-2 family) for early diagnosis of the disease, and to understand its pathogenesis as a first step in improving methods of treatment. The discovery of these potential biomarkers and molecular targets for cancer diagnostics and therapeutics has the potential to significantly change the clinical approach and outcome of the disease.

On the Milled Wood Lignins Isolated from Hardwood by Progressive Milling (단계적(段階的) 분쇄법(粉碎法)에 의해 조제(調製)된 활엽수(闊葉樹) MWL에 관한 연구(硏究))

  • Cho, Nam Seok
    • Journal of Korean Society of Forest Science
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    • v.45 no.1
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    • pp.62-67
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    • 1979
  • Ultraviolet microscopy of ultrathin sections of wood has proved to be one of the useful means for determining the lignin distribution in the various regions of the cell wall. Also, spectral approach and quantitative analysis of isolated compound middle lamella fraction from birch xylem have revealed that the lignin associated with the vessel secondary wall and middle lamella is composed predominantly of gualacylpropane units. Lignin deposited in the fiber and ray parenchyma secondary walls is composed mostly of syringylpropane units. The middle lamella lignin around fibers and ray cells contains both guaiacyl and syringyl propane quits. On the basis of the results above, this research was carried out to clarify the origin of milled wood lignin (MWL) by analysing the chemical characteristics of ML MWLs extracted at various milling stages. The amount of phenolic hydroxyl-, ${\alpha}$-carbonyl-, and methoxyl-group in the MWL's increases the milling time. And progressive mining contributes to the merease of ratio of syringylaldehyde to vanillin(S/V ratio) after nitrobenzene oxidation of MWL. Accordingly, It could be concluded that milled wood lignin extracted at the initial milling stage derives from compound middle lamella region of cell wall, whereas, with progressive milling, lignin of secondary wall of fiber is introduced gradually to milled wood lignin. These results are suggesting that heterogeneous chemical structure of lignins in hardwood exists. Although milled wood lignin at the initial stage seems to have lower molecular weight in comparison with milled wood lignin extracted at final milling stage from the result of Gel-filtration curves, further study would be required on molecular weight distribution of milled wood lignin in future.

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Comparison of Results between Cytogenetic Technique and Molecular Genetic Technique in Colorectal Carcinoma Patients (대장암환자의 염색체 결실에서 세포유전학적 기법과 분자유전학적 기법의 결과 비교)

  • Park, Cheolin;Lee, Jae Sik
    • Korean Journal of Clinical Laboratory Science
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    • v.49 no.3
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    • pp.285-293
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    • 2017
  • Globally, 1.3 million people develop colon cancer every year, and 600,000 people die each year it. In Korea, colorectal carcinoma was associated with the highest death rate, accounting for 8,380 people, among solid cancers in 2015. Among the various methods for the diagnosis and study of colorectal carcinoma, the results obtained by cytogenetic and molecular genetic methods were compared. Detection rate was 47% in 18q, 40% in 17p, 27% in 22q, and 17% in 10q via CGH; detection rate was 57% in D18S59, 50% in D18S68, 50% in TP53CA, 47% in D18S6940% in D22S274, 37% in D22S283, 27% in D10S187, and 23% in D10S541 with LOH. Microsatellite marker matching rates were 100% in D22S274, 100% in D22S283, 100% in D10S186, 100% in D10S187, 100% in D10S541, 93% in D18S69, 93% in D18S68, 92% in TP53CA, and 89% in D18S59. The agreement rate between the two methods was 94.4% based on positive results using CGH. Based on the advantages of CGH, which was the ability to obtain information regarding the entire tumor genome at once, this experiment could identify the region with significant deletion using CGH and the more limited region LOH, with a completely different approach. LOH in the recurrent high-risk group, 18q21, was helpful in the selection of treatment modalities and in prognostic estimation as well as making the most appropriate decision for treatment. Therefore, it is suggested that LOH with surgical site tissues could be one of the treatment methods for recurrent high-risk group among patients with colorectal carcinoma.

Characterization of Bovine Lymphocyte Antigen DRB3 exon2 Gene of Korean Native Cattle (한우의 BoLA DRB3 exon2 유전자의 특성)

  • Kang, Ho Bum;Ryoo, Seung Heui;Lee, Sang Hoon;Jeon, Byung Soon;Sang, Byung Chan
    • Korean Journal of Agricultural Science
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    • v.25 no.1
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    • pp.79-88
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    • 1998
  • This study was performed in order to apply to effective breeding of Korean native cattle on the molecular genetic level obtained from PCR and nucleotide sequencing analysis of BoLA DRB3 exon2 that has important roles in host immune defence. Genomic DNA used in this study was prepared from the blood of Korean native cattle in Korean Native Cattle Improvement Center of National Livestock Cooperation. The results obtained from this study are summarized as follows: 1. Genomic DNA extracted from the blood of Korean native cattle was subjected to electrophoresis on 1.5% agarose gel. Major band was bigger than 12.2kb, indicating that genomic DNA was well prepared for PCR. Amplified products of 284bp fragments was obtained the amplification of BoLA DRB3 exon2 gene by PCR. 2. Cloning of BoLA DRB3 exon2 of Korean native cattle with pCR2.1 vector was conformed by 300bp fragment from recombinent plasmid that restricted with enzyme digestion of EcoRI. 3. Homology of BoLA DRB3 exon2 alleles of parent was 82.0% between sire's alleles and 90.1% between dam's alleles. 4. In pedigree analysis using BoLA DRB3 exon2 gene, sequencing result of BoLA DRB3 exon2 genes showed inheritance by Mendelian mode through the parents to their offspring. 5. Taking together those experimental results, pedigree was confirmed on the basis of sequencing for the alleles of parents and offspring. This knowledge by the molecular biological approach could be served for the improvement of Korean native cattle.

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Physiological and molecular characterization of two inbred radish lines with different bolting times (추대시기가 서로 다른 무 계통간 생리학적, 분자생물학적 개화 특성 규명)

  • Park, Hyun Ji;Jung, Won Yong;Lee, Sang Sook;Lee, Joo won;Kim, Youn-Sung;Cho, Hye Sun
    • Journal of Plant Biotechnology
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    • v.42 no.3
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    • pp.215-222
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    • 2015
  • The radish (Raphanus sativus L.) is an important Brassicaceae root vegetable crop worldwide. Several studies have been conducted concerning radish breeding. There are major challenges to prevent premature bolting in spring plantings. Here, we performed the characterization of two inbred radish lines which vary in bolting time. "Late bolting radish" (NH-JS1) and "early bolting radish" (NH-JS2) were generated by a conventional breeding approach. The two inbred lines showed different bolting phenotypes depending on vernalization time at $4^{\circ}C$. NH-JS1, the late bolting radish, was less sensitive to cold treatment and the less sensitivity was inversely proportional to the duration of the vernalization. We also measured gene expression levels of the major bolting time related genes in the NH-JS1 and NH-JS2 lines. RsFLC1 plays a central role in the timing of flowering initiation. It is a strong repressor and it's transcript is highly expressed in NH-JS1 compared to NH-JS2 under no treatment and vernalization conditions. RsFRI, a positive regulator of RsFLC, is also highly expressed in NH-JS1 compared to NH-JS2 regardless of vernalization. In contrast, RsSOC1, suppressed by FLC as a floral integrator gene, showed the most difference, a 5-fold increase, between NH-JS1 and NH-JS2 under vernalization conditions. From these results, we conclude that NH-JS1 showed a late flowering phenotype after cold treatment due to the expression differences of flowering time regulator genes rather than difference sensitivity to cold. These results may be useful to understand the control mechanisms of flowering time and may help identify molecular markers for selecting late bolting trait in radish.

Overview of Real-time Visibility System for Food (Livestock Products) Transportation Systems on HACCP Application and Systematization (축산물 유통단계의 HACCP 적용과 체계화를 위한 실시간 관제시스템에 대한 현황)

  • Kim, Hyoun-Wook;Lee, Joo-Yeon;Hong, Wan-Soo;Hwang, Sun-Min;Lee, Victor;Rhim, Seong-Ryul;Paik, Hyun-Dong
    • Food Science of Animal Resources
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    • v.30 no.6
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    • pp.896-904
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    • 2010
  • HACCP is a scientific and systematic program that identifies specific hazards and gives measurements in order to control them and ensure the safety of foods. Transportation of livestock and its products is one of the vulnerable sectors regarding food safety in Korea, as meats are transported by truck in the form of a carcass or packaged meat in a box. HACCP application and its acceleration of distribution, in particular transportation, are regarded as important to providing consumers with ultimately safe livestock products. To achieve this goal, practical tools for HACCP application should be developed. Supply chain management (SCM) is a holistic and strategic approach to demand, operations, procurement, and logistics process management. SCM has been beneficially applied to several industries, notably in vehicle manufacture and the retail trade. HACCP-based real-time visibility system using wireless application (WAP) of the livestock distribution is centralized management system that enables control of temperature and HACCP management in real-time for livestock transportation. Therefore, the application of HACCP to livestock distribution (transportation, storage, and sale) can be activated. Using this system, HACCP management can be made easier, and distribution of safe livestock products can be achieved.

Variation in bioactive principles and bioactive compounds of Rosa rugosa fruit during ripening (해당화 열매 성숙단계에 따른 생리활성 및 기능성 물질 변화 분석)

  • Kwak, Minjeong;Eom, Seung Hee;Gil, Jinsu;Kim, Ju-Sung;Hyun, Tae Kyung
    • Journal of Plant Biotechnology
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    • v.46 no.3
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    • pp.236-245
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    • 2019
  • Fruit ripening is a genetically programmed process involving a number of biochemical and physiological processes assisted by variations in gene expression and enzyme activities. This process generally affects the phytochemical profile and the bioactive principles in fruits and vegetables. To appraise the variation in bioactive principles of fruits from Rosa rugosa during its ripening process, we analyzed the changes in antioxidant and anti-elastase activities and polyphenolic compounds during the four ripening stages of fruits. Overall, an extract of unripe fruits contained the highest levels of total phenolic and flavonoid contents, radical scavenging activity, reducing power, oxygen radical antioxidant capacity, and elastase inhibitory activity, compared with the extracts of fruits at other stages of ripening. Additionally, we found that the reduction of flavonoid content occurs because of decreased transcriptional levels of genes involved in flavonoid biosynthesis pathway during the ripening process. Based on HPLC analysis, we found that the extract of unripe fruits contained the highest amount of myricetin, caffeic acid, chlorogenic acid, syringic acid, and p-coumaric acid and suggested that the antioxidant and anti-elastase activities of the extract obtained from stage 1, should be mediated by the presence of these compounds. Additionally, we analyzed the interaction sites and patterns between these compounds and elastase using the structure-based molecular docking approach, and suggested that chlorogenic acid strongly interacted with elastase. Together, these findings suggest that the maturity of fruits has profound effects on the pharmaceutical value of R. rugosa.

THE PREVALENCE OF WHITE SPOT LESIONS ON THE MESIAL SURFACES OF THE 1ST MOLARS IN CHILDREN AND MINIMAL INVASIVE APPROACH - A PILOT STUDY (어린이 제1대구치 근심면 초기 우식의 유병률과 최소 침습적 접근)

  • Ahn, Myung-Ki;Lee, Geum-Lang;Jeong, Tae-Sung;Kim, Shin
    • Journal of the korean academy of Pediatric Dentistry
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    • v.36 no.1
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    • pp.102-107
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    • 2009
  • In clinical pediatric dentistry, we have many chances to encounter the white spot like incipient enamel lesions on the mesial surfaces of the 1st molars with direct vision, especially just after the 2nd primary molars were exfoliated. But it was thought highly desirable to assess if these lesions are properly and effectively managed yet. This study aims at surveying the prevalence of incipient lesions on the mesial surfaces of the 1st molars in children through direct observation and examining the suitability of adhesive sealing on them as a pilot trial in searching for their proper management. 1. Among the 124 mesial surfaces of the 1st molars examined, 34% were sound, 53% had incipient carious lesions and 13% had cavitated lesions. 2. In the sectional views of the specimens, 20% showed microleakage after thermo-cycling and it was thought not recommendable as a permanent method. Therefore in order to effectively fight against the incipient caries lesions in children‘s permanent teeth, it was thought proper not to rely on any one method, but to perform reinforcing oral hygiene and promotion of remineralization in combination with therapeutic sealing which is stronger in short-term sealing effect. Although therapeutic sealing has been considered as the core in minimally invasive concept to treat the white spot lesions, its long-term clinical trials have not been suggested. Continuous research is strongly required for making this approach to acquire permanent nature, especially in regards of proper pretreatment and high molecular materials deeply penetrable into enamel.

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Pharmacological Comparison of Timosaponin A III on the 5-beta Reductase and Androgen Receptor via In Silico Molecular Docking Approach (In silico 약리학적 분석을 통한 티모사포닌 A III의 5-베타 리덕타아제 단백질 및 안드로겐 수용체 단백질 활성 부위에 대한 결합 친화도 비교 연구)

  • Kim, Dong-Chan
    • Journal of Life Science
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    • v.28 no.3
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    • pp.307-313
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    • 2018
  • Alopecia cause psychological stress due to their effect on appearance. Thus, the global market size of the alopecia treatment products are growing quickly. Timosaponin A III is the well known active ingredient of Anemarrhenae Rhizoma. In this study, we investigated and compared the binding affinity of timosaponin A III with finasteride (5-beta reductase antagonist) and minoxidil (androgen receptor antagonist) on the target protein active site by in silico computational docking studies. The three dimensional crystallographic structure of 5-beta reductase (PDB ID : 3G1R) and androgen receptor (PDB ID: 4K7A) was obtained from PDB database. In silico computational autodocking analysis was performed using PyRx, Autodock Vina, Discovery Studio Version 4.5, and NX-QuickPharm option based on scoring functions. The timosaponin A III showed optimum binding affinity (docking energy) with 5-beta reductase as -12.20 kcal/mol as compared to the finasteride (-11.70 kcal/mol) and with androgen receptor as -9.00 kcal/mol as compared to the minoxidil (-7.40 kcal/mol). The centroid X, Y, Z grid position of the timosaponin A III on the 5-beta reductase was similar (overlap) to the finasteride, but the X, Y, Z centroid grid of the timosaponin A III on the androgen receptor was significantly far from the minoxidil centroid position. These results significantly indicated that timosaponin A III could be more potent antagonist to the 5-beta reductase and androgen receptor. Therefore, the extract of Anemarrhenae Rhizoma or timosaponin A III containing biomaterials can substitute the finasteride and minoxidil and can be applied to the alopecia protecting product and related industrial fields.