• Title/Summary/Keyword: Alternative protein

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Aequorin Based Functional Assessment of the Melanin Concentrating Hormone Receptor by Intracellular Calcium Mobilization

  • Lee, Sung-Hou
    • Biomolecules & Therapeutics
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    • v.18 no.2
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    • pp.152-158
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    • 2010
  • Melanin concentrating hormone is a neuropeptide highly expressed in the brain that regulates several physiological functions mediated by receptors in the G-protein coupled receptor family, especially plays an important role in the complex regulation of energy balance and body weight mediated by the melanin concentrating hormone receptor subtype 1 (MCH1). Compelling pharmacological evidence implicating MCH1 signaling in the regulation of food intake and energy expenditure has generated a great deal of interest by pharmaceutical companies as MCH1 antagonists may have potential therapeutic benefit in the treatment of obesity and metabolic syndrome. Although fluorescence-based calcium mobilization assay platform has been one of the most widely accepted tools for receptor research and drug discovery, fluorescence interference and shallow assay window limit their application in high throughput screening and have led to a growing interest in alternative, luminescence-based technologies. Herein, a luminescence-based functional assay system for the MCH1 receptor was developed and validated with the mitochondrial targeted aequorin. Aequorin based functional assay system for MCH1 presented excellent Z' factor (0.8983) and high signal-to-noise ratio (141.9). The nonpeptide MCH1 receptor antagonist, SNAP 7941 and GSK 803430, exhibited $IC_{50}$ values of 0.62 ${\pm}$ 0.11 and 12.29 ${\pm}$ 2.31 nM with excellent correlation coefficient. These results suggest that the aequorin based assay system for MCH1 is a strong alternative to the traditional GPCR related tools such as radioligand binding experiments and fluorescence functional determinations for the compound screening and receptor research.

Effects of Rape Residue on Nitrogen Fertilizer Reduction in Paddy Soil under Double Cropping System

  • Cho, Hyun-Jun;Hyun, Byung-Keun;Sonn, Yeon-Kyu;Shin, Kook Sig
    • Korean Journal of Soil Science and Fertilizer
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    • v.49 no.1
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    • pp.7-11
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    • 2016
  • Winter crops have been recognized as an alternative to soil management for fertility and crop productivity in paddy soil. Recently, rape has been produced at winter season and there is little research results on reduction of N fertilizer by adding rape residues for rice cultivation. In this study, we investigated the productivity and quality of rice by applying with 0, 27, 63, $90kg\;ha^{-1}$ of N fertilizer after input of rape residues into soil for two years. The highest yield of rice was average $4.68Mg\;ha^{-1}$ at the treatment applied with the $90kg\;N\;ha^{-1}$ with rape residue. It was reduced to N rate from 31.6 to $43.2kg\;N\;ha^{-1}$ in comparison to $4.53Mg\;ha^{-1}$ of maximum yield in treatment added $90kg\;N\;ha^{-1}$ with rape. Amylose content in rice was similar among treatments with/without rape residue, but protein content was lower in treatment with rape residue than in those without rape residue. In conclusion, input of rape residue in rice cultivation could be alternative to reduction of N fertilization and improvement of quality by adjusting rice productivity in paddy soil under cropping system.

Alternative Splicing Pattern Analysis from RNA-Seq data (RNA-Seq 데이터를 이용한 선택 스플라이싱 유형 분석)

  • Kong, Jin-Hwa;Lee, Jong-Keun;Lee, Un-Joo;Yoon, Jee-Hee
    • Proceedings of the Korean Information Science Society Conference
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    • 2011.06a
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    • pp.37-40
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    • 2011
  • 선택 스플라이싱 (alternative splicing)은 mRNA (messenger RNA)의 전구체인 pre-mRNA가 mRNA로 전사될 때 pre-mRNA의 엑손 영역들 (exons)이 여러 가지 유형 (pattern)으로 다시 연결되는 과정을 말한다. 선택 스플라이싱에 의해 하나의 유전자로부터 서로 다른 mRNA가 만들어 지고 서로 다른 이소형의 단백질 (protein isoforms)이 생성된다. 현재까지 알려진 선택 스플라이싱의 유형은 약 7가지 종류가 있으며, 유전자의 돌연변이 및 질병과 밀접한 연관성을 가지고 있는 것으로 알려져 있다. 본 연구에서는 차세대 시퀀싱 (Next Generation Sequencing : NGS) 기술로 생성된 RNA-Seq 데이터로부터 각 유전자 영역에 대한 선택 스플라이싱 유형을 분류/추출하는 새로운 알고리즘을 제안한다. 제안된 알고리즘에서는 RNA-Seq 데이터를 DNA 시퀀스와 mRNA 트랜스크립트 시퀀스에 동시 매핑하고, 각 엑손 영역에 정렬된 RNA-Seq 데이터의 커버리지 정보 및 엑손의 접합 (junction) 정보를 이용하여 발현된 트랜스크립트 (transcript)의 종류와 양을 측정한다. 알고리즘의 유효성을 보이기 위하여 시뮬레이션 데이터를 이용한 인간 유전자 영역에서의 선택 스플라이싱 유형 추출 실험을 수행하였으며, 검증된 선택 스플라이싱 DB와 비교, 검증하였다.

Non-Conventional Concentrates in Temperate Asian-Australasian Countries - Review -

  • Chiou, P.W.S.
    • Asian-Australasian Journal of Animal Sciences
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    • v.12 no.3
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    • pp.460-466
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    • 1999
  • The huge amount of demand for feedgrains from this region could not possibly be met by producing countries from the other regions. In order to fulfill this increasing demand for conventional raw materials, an alternative for the conventional raw materials produced in the Asia and Pacific region is becoming increasingly more important. A potential alternative is concentrates or non-conventional concentrates produced locally in relative abundance in this region. These feedstuffs include feed grains, by-products from the milling, sugar industries, brewing and distilling industries. Vegetable, citrus, and animal by-products from abattoir, feather meal and blood meal are also possibilities. In addition to more widespread use of unconventional feed sources, the following approach is recommended to improve utilization and performance. These include establishing the nutritive value of non-conventional feeds, quality control to minimize variability, proper storage and processing to assure the nutritive value and prevent mycotoxin contamination, properly balance amino acids with protein sources, supplementation with synthetic amino acids and the use of enzymes to increase digestibility. Currently, practical applications for these resources in feed formulation are negligible despite the potential. The socio-economic aspects will dominate the use of these non-conventional concentrates. In the future, the feed industry will resolve the problems in using locally available raw feed materials.

Acidifier as an Alternative Material to Antibiotics in Animal Feed

  • Kim, Y.Y.;Kil, D.Y.;Oh, H.K.;Han, In K.
    • Asian-Australasian Journal of Animal Sciences
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    • v.18 no.7
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    • pp.1048-1060
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    • 2005
  • Dietary acidifiers appear to be a possible alternative to feed antibiotics in order to improve performance of weaning pigs. It is generally known that dietary acidifiers lower gastric pH, resulting in increased activity of proteolytic enzymes, improved protein digestibility and inhibiting the proliferation of pathogenic bacteria in GI tract. It is also hypothesized that acidifiers could be related to reduction of gastric emptying rate, energy source in intestine, chelation of minerals, stimulation of digestive enzymes and intermediate metabolism. However, the exact mode of action still remains questionable. Organic acidifiers have been widely used for weaning pigs' diets for decades and most common organic acidifiers contain fumaric, citric, formic and/or lactic acid. Many researchers have observed that dietary acidifier supplementation improved growth performance and health status in weaning pigs. Recently inorganic acidifiers as well as organic acidifiers have drawn much attention due to improving performance of weaning pigs with a low cost. Several researchers introduced the use of salt form of acidifiers because of convenient application and better effects than pure state acids. However, considerable variations in results of acidifier supplementation have been reported in response of weaning pigs. The inconsistent responses to dietary acidifiers could be explained by feed palatability, sources and composition of diet, supplementation level of acidifier and age of animals.

Alternative Isoforms of TonEBP with Variable N-termini are Expressed in Mammalian Cells

  • Kim, Hyo-Shin;Son, Sook-Jin;Kim, Seon-Nyo;Kim, Yong-Duk;Kim, Kwang-Jin;Jeon, Byeong-Hwa;Park, Jin-Bong;Lee, Sang-Do
    • The Korean Journal of Physiology and Pharmacology
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    • v.11 no.3
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    • pp.135-138
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    • 2007
  • Hypertonicity imposes a great deal of stress to cells since it causes rise in cellular ionic strength, which can be reduced by the accumulation of compatible osmolytes. TonEBP plays a central role in the cellular accumulation of compatible osmolytes via transcriptional stimulation of membrane transporters and aldose reductase. Alternatively spliced forms of TonEBP mRNA have previously been reported and two of them showed different transcriptional activity. In the present study, isoform-specific antibodies were produced to confirm the translation of the spliced mRNA to protein. TonEBP was immunoprecipitated by using anti-TonEBP antibody and then immunoblotted using anti-TonEBP or isoform specific antibodies to find out the expression profile of TonEBP isoforms in basal or stimulated condition. From these results, we conclude that all TonEBP isoforms are expressed in mammalian cells and their expression patterns are not same in every cells.

Use of Probiotics in Dairy Industry to Improve Productivity and as an Alternative to Antibiotics (낙농산업에서 항생제 사용의 문제점과 프로바이오틱스의 활용을 통한 생산성 향상)

  • Seo, Yeongeun;Yoo, Yoonjeong;Yoon, Yohan
    • Journal of Dairy Science and Biotechnology
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    • v.39 no.2
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    • pp.63-67
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    • 2021
  • Antibiotics are widely used to improve productivity in the dairy industry. However, the inappropriate use of antibiotics causes the deterioration in the quality of dairy products undergoing fermentation and maturation. Hence, probiotic use is emerging as an alternative to curb the increased utilization of antibiotics. Probiotics are defined as "living microorganisms that, when administered in appropriate amounts, confer health benefits on the host." They may improve host disease resistance by regulating intestinal microflora balance and promote animal growth and development. In the dairy industry, probiotics have been studied to increase milk production by improving digestion in dairy cows, enhance the content of dairy components such as milk fat and protein, reduce the risk of mastitis in cows, and increase calf weight. Thus, the use of probiotics can improve the production and safety of dairy products. However, some probiotics are still unstable during storage and have low quality and safety issues. Therefore, to reduce the use of antibiotics in the dairy industry, probiotics should be developed and produced considering the above-mentioned problems.

Screening and isolation of antibacterial proteinaceous compounds from flower tissues: Alternatives for treatment of healthcare-associated infections

  • de Almeida, Renato Goulart;Silva, Osmar Nascimento;de Souza Candido, Elizabete;Moreira, Joao Suender;Jojoa, Dianny Elizabeth Jimenez;Gomes, Diego Garces;de Souza Freire, Mirna;de Miranda Burgel, Pedro Henrique;de Oliveira, Nelson Gomes Junior;Valencia, Jorge William Arboleda;Franco, Octavio Luiz;Dias, Simoni Campos
    • CELLMED
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    • v.4 no.1
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    • pp.5.1-5.8
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    • 2014
  • Healthcare-associated infection represents a frequent cause of mortality that increases hospital costs. Due to increasing microbial resistance to antibiotics, it is necessary to search for alternative therapies. Consequently, novel alternatives for the control of resistant microorganisms have been studied. Among them, plant antimicrobial protein presents enormous potential, with flowers being a new source of antimicrobial molecules. In this work, the antimicrobial activity of protein-rich fractions from flower tissues from 18 different species was evaluated against several human pathogenic bacteria. The results showed that protein-rich fractions of 12 species were able to control bacterial development. Due its broad inhibition spectrum and high antibacterial activity, the protein-rich fraction of Hibiscus rosa-sinensis was subjected to DEAE-Sepharose chromatography, yielding a retained fraction and a non-retained fraction. The retained fraction inhibits 29.5% of Klebsiella pneumoniae growth, and the non-retained fraction showed 31.5% of growth inhibition against the same bacteria. The protein profile of the chromatography fractions was analyzed by using SDS-PAGE, revealing the presence of two major protein bands in the retained fraction, of 20 and 15 kDa. The results indicate that medicinal plants have the biotechnological potential to increase knowledge about antimicrobial protein structure and action mechanisms, assisting in the rational design of antimicrobial compounds for the development of new antibiotic drugs.

Binding of the His-tagged Tail Protein J of Bacteriophage Lambda with Escherichia coli K-12 (히스티딘으로 표지된 람다 박테리오파아지 꼬리 단백질 J와 대장균 K-12와의 결합)

  • Shin, Hae Ja
    • Journal of Life Science
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    • v.28 no.1
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    • pp.78-82
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    • 2018
  • Detection of pathogenic microorganisms takes several days by conventional methods. It is necessary to assess microorganisms in a timely manner to reduce the risk of spreading infection. For this purpose, bacteriophages are chosen for use as a biosensing tool due to their host specificity, wide abundance, and safety. However, their lytic cycle limits their efficacy as biosensors. Phage proteins involved in binding to bacteria could be a robust alternative in resolving this drawback. Here, a fragment of tail protein J (residues 784 to 1,132) of phage lambda fused with 6X His-tag (6HN-J) at its N-terminus was cloned, overexpressed, purified, and characterized for its binding with microorganisms. The purified protein demonstrated a size of about 38 kDa in sodium dodecyl sulfate - polyacrylamide gel electrophoresis (SDS-PAGE) and bound with anti-His monoclonal antibodies. It bound specifically to Escherichia coli K-12, and not Salmonella typhimurium, Bacillus subtilis, or Pseudomonas aeruginosa in dot blotting. Binding of the protein to E. coli K-12 inhibited about 50% of the in vivo adsorption of the phage lambda to host cells at a concentration of $1{\mu}g/ml$ 6HN-J protein and almost 100% at $25{\mu}g/ml$ 6HN-J. The results suggest that a fusion viral protein could be utilized as a biosensing element (e.g., protein chips) for detecting microorganisms in real time.

Evaluation of Feed Value of Barley Fodder as an Alternative Feed Ingredient

  • Kim, Tae-Il;Mayakrishnan, Vijayakumar;Lim, Dong-Hyun;Lee, Hyun-Jeong;Son, Jun-Kyu;Kim, Yoon-Jeong;Choi, Hee-Chul;Shin, Jae-Hyeong;Park, Jong-Ho;Kim, Sam-Churl;Ki, Kwang-Seok
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.40 no.3
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    • pp.161-166
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    • 2020
  • Barley is an important cereal gain which is traditionally used in some nations of Asia and North Africa, and there has been growing interest in using barley as an ingredient in food due to their nutritional value and high content of phyto-constituents. However, no study report on comparative feed value between sprouted barley, cornflake and alfalfa hay. Therefore, in this study we aimed to evaluate the chemical composition, amino acid profile and mineral content of 6 day sprouted barley fodder (SBF) compared with cornflake and alfalfa hay using by AOAC method, as an alternative feed ingredient. Results showed that SBF had higher content of crude protein, acid detergent insoluble crude proteins and neutral detergent insoluble crude protein than alfalfa hay and cornflake; cornflake had higher crude fiber, neutral detergent fiber content than SBF and alfalfa hay; alfalfa hay had higher crude fiber, crude ash, acid detergent fiber, neutral detergent fiber and lignin level than SBF and cornflake. Also, significant differences were found on amino acid content among them (p<0.01). The most abundant amino acid in SBF was glutamate (123 g/kg DM), which is higher than in alfalfa hay (1.27%) or cornflake (1.58%). However, methionine (1.33%) and cysteine (1.53%) were the least abundant amino acids in SBF compared with cornflake or alfalfa hay. Furthermore, our study results exhibited that SBE comprise a good sources of minerals including ferrous (90.01 mg/kg) followed by zinc (20.50 mg/kg), magnesium (0.20 mg/kg) and sodium (0.03 mg/kg) as compared to cornflake and alfalfa hay. The present research findings, confirmed that the nutritional values of SBF are comparable to those of cornflake and alfalfa hay. Hence, SBF can be a better alternative feed ingredient for cornflake or alfalfa hay. However, feeding trials will be required to determine acceptability of SBF for ruminant production.