• Title/Summary/Keyword: Pretreatment temperature

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Integrated Rotary Genetic Analysis Microsystem for Influenza A Virus Detection

  • Jung, Jae Hwan;Park, Byung Hyun;Choi, Seok Jin;Seo, Tae Seok
    • Proceedings of the Korean Vacuum Society Conference
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    • 2013.08a
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    • pp.88-89
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    • 2013
  • A variety of influenza A viruses from animal hosts are continuously prevalent throughout the world which cause human epidemics resulting millions of human infections and enormous industrial and economic damages. Thus, early diagnosis of such pathogen is of paramount importance for biomedical examination and public healthcare screening. To approach this issue, here we propose a fully integrated Rotary genetic analysis system, called Rotary Genetic Analyzer, for on-site detection of influenza A viruses with high speed. The Rotary Genetic Analyzer is made up of four parts including a disposable microchip, a servo motor for precise and high rate spinning of the chip, thermal blocks for temperature control, and a miniaturized optical fluorescence detector as shown Fig. 1. A thermal block made from duralumin is integrated with a film heater at the bottom and a resistance temperature detector (RTD) in the middle. For the efficient performance of RT-PCR, three thermal blocks are placed on the Rotary stage and the temperature of each block is corresponded to the thermal cycling, namely $95^{\circ}C$ (denature), $58^{\circ}C$ (annealing), and $72^{\circ}C$ (extension). Rotary RT-PCR was performed to amplify the target gene which was monitored by an optical fluorescent detector above the extension block. A disposable microdevice (10 cm diameter) consists of a solid-phase extraction based sample pretreatment unit, bead chamber, and 4 ${\mu}L$ of the PCR chamber as shown Fig. 2. The microchip is fabricated using a patterned polycarbonate (PC) sheet with 1 mm thickness and a PC film with 130 ${\mu}m$ thickness, which layers are thermally bonded at $138^{\circ}C$ using acetone vapour. Silicatreated microglass beads with 150~212 ${\mu}L$ diameter are introduced into the sample pretreatment chambers and held in place by weir structure for construction of solid-phase extraction system. Fig. 3 shows strobed images of sequential loading of three samples. Three samples were loaded into the reservoir simultaneously (Fig. 3A), then the influenza A H3N2 viral RNA sample was loaded at 5000 RPM for 10 sec (Fig. 3B). Washing buffer was followed at 5000 RPM for 5 min (Fig. 3C), and angular frequency was decreased to 100 RPM for siphon priming of PCR cocktail to the channel as shown in Figure 3D. Finally the PCR cocktail was loaded to the bead chamber at 2000 RPM for 10 sec, and then RPM was increased up to 5000 RPM for 1 min to obtain the as much as PCR cocktail containing the RNA template (Fig. 3E). In this system, the wastes from RNA samples and washing buffer were transported to the waste chamber, which is fully filled to the chamber with precise optimization. Then, the PCR cocktail was able to transport to the PCR chamber. Fig. 3F shows the final image of the sample pretreatment. PCR cocktail containing RNA template is successfully isolated from waste. To detect the influenza A H3N2 virus, the purified RNA with PCR cocktail in the PCR chamber was amplified by using performed the RNA capture on the proposed microdevice. The fluorescence images were described in Figure 4A at the 0, 40 cycles. The fluorescence signal (40 cycle) was drastically increased confirming the influenza A H3N2 virus. The real-time profiles were successfully obtained using the optical fluorescence detector as shown in Figure 4B. The Rotary PCR and off-chip PCR were compared with same amount of influenza A H3N2 virus. The Ct value of Rotary PCR was smaller than the off-chip PCR without contamination. The whole process of the sample pretreatment and RT-PCR could be accomplished in 30 min on the fully integrated Rotary Genetic Analyzer system. We have demonstrated a fully integrated and portable Rotary Genetic Analyzer for detection of the gene expression of influenza A virus, which has 'Sample-in-answer-out' capability including sample pretreatment, rotary amplification, and optical detection. Target gene amplification was real-time monitored using the integrated Rotary Genetic Analyzer system.

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Effects on the Storage Life of Satsuma Mandarin by the Pretreatment at various Temperatures (저장전 온도처리가 온주밀감의 저장에 미치는 영향)

  • Kim, Wan-Taek;Lee, Sang-Yong;Kim, Ji-Yong;Kang, Chang-Hee;Koh, Jeong-Sam
    • Applied Biological Chemistry
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    • v.41 no.3
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    • pp.228-233
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    • 1998
  • The storage life of satsuma mandarin(Citrus unshiu Marc. var. miyagawa) by the various pretreatment of temperatures; non-treated, room temperature, $10,\;20^{\circ}C$ and $35^{\circ}C$ were investigated. The pretreated citrus fruits were stored at $4^{\circ}C$, 85% relative humidity. Weight loss of citrus fruits by the pretreatment at $35^{\circ}C$ for 24 hrs was the lowest among that of others. Decay ratio of $35^{\circ}C$ pretreated fruits was increased at initial stages of storage, but was maintained low level after that, compared to other treatments. After 115 days storage, firmness of fruits was lowered by the softening, and decayed fruits were occurred increasingly. Ethylene evolution was increased between $55{\sim}65$ days after storage, and the amount was increasing rapidly after 115 days. It seemed to be derived from decayed fruits and physiological activities. $CO_2$ content in fruit was decreased at initial stages of storage, but was increased between $55{\sim}100$ days during storage periods. Acid content, soluble solids, total sugar and vitamin C were reduced gradually during cold storage, but the difference among treatments was not so great. Pretreated fruits at $35^{\circ}C$ for 24 hrs before cold storage was effective on preventing from weight loss and respiration ratio. Optimum storage period of early variety of Satuma mandarin was regarded for 100 days on the basis of appeareance and taste.

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Study on the Pretreatment of Rice Hull to Enhance Enzymatic Saccharification Efficiency (효소 당화효율 증진을 위한 왕겨의 전처리 방법 연구)

  • Bark, Surn-Teh;Koo, Bon-Cheol;Moon, Youn-Ho;Cha, Young-Lok;Yoon, Young-Mi;Kim, Jung Kon;An, Gi Hong;Park, Kwang-Geun;Park, Don-Hee
    • Applied Chemistry for Engineering
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    • v.23 no.4
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    • pp.399-404
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    • 2012
  • The objective of this study was to investigate the efficient pretreatment method for bioethanol production from rice hull. Ammonia and sodium hydroxide as an alkaline solution and dilute sulfuric acid as an acidic solution were used in a batch reactor under high-temperature and high-pressure conditions. The highest enzymatic saccharification efficiency of 82.8% and ash removal rate of 94.7% were obtained in the dilute sulfuric acid treated sample after the sodium hydroxide solution treatment. The enzymatic saccharification efficiencies and ash removals of pretreated rice hull samples have very similar variation tendency. This means that the maximum obstructive factor for the enzymatic saccharification of rice hull is the ash (silicate) content in biomass. The findings suggest that the combined sodium hydroxide-dilute sulfuric acid treatment system under high-temperature and high-pressure conditions is a promising pretreatment method to enhance the enzymatic saccharification of the silica-rich biomass.

A Manufacturing Technique of Agar with Strong Gelling Ability from Gelidium amansii (우뭇가사리로부터 고강도 한천의 제조)

  • DO Jeong-Ryong;PARK Jin-Hee;JO Kil-Suk
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.31 no.5
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    • pp.673-676
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    • 1998
  • Preparative conditions of high-gel strength agar from Gelidium amansii have been studied, The effect of NaOH pretreatment on the quality and yields of agar extracted from Gelidium amansli was examined. The Bel strength of agar extracted from C. amansii pretreated with NaOH was higher than that of agar extracted from G. amansii non-pretreated with NaOH. The gel strength of agar extracted from G. amansii was influenced by concentration, temperature and time of pretreatment with NaOH. It was found that the proper concentration, temperature and time of NaOH pretreatment to produce high-gel strength agar was $6\%$ NaOH, $80^{\circ}C$ and 2$\~$3 hrs. The principal sugars of agar extracted from G. amansli were galactose and 3,6-anhydrogalactose.

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Development of a Temperature Controller for Microwave-assisted Digestion System for Agricultural Samples (농식품 시료 전처리를 위한 마이크로웨이브 분해기용 온도 제어장치 개발)

  • Mo, Chang-Yeon;Kim, Gi-Young;Kim, Hak-Jin;Kim, Yong-Hun;Yang, Kil-Mo;Lee, Kang-Jin
    • Journal of Biosystems Engineering
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    • v.34 no.5
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    • pp.371-376
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    • 2009
  • Microwave digestion is a preferred pretreatment method for agricultural samples because of its quick chemical reaction and minimum loss of analytes. In this research, a feedback temperature controller was developed to control the temperature inside a vessel for the microwave-assisted digestion system. An existing industrial microwave oven was fitted with the temperature controller for controlling inside temperature of the vessel. Four control methods, On/Off, proportional (P), proportional integral (PI), and proportional integral derivative (PID) were used and compared. Experimental results showed that PID control produced best temperature control performance. The PID controller could maintain the temperature of water sample and rice sample in the digestion system with error range of $-2.5{\sim}3.3^{\circ}C$ and $-1.9{\sim}0.5^{\circ}C$ at set temperature of $170^{\circ}C$, respectively.

Effect of Soluble Starch Pretreatment and Storage Condition on Caking Degree and Moisture Sorption of Powdered Onion (가용성 전분의 전처리와 저장조건이 분말양파의 Caking과 수분흡수에 미치는 효과)

  • 김명환
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.20 no.3
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    • pp.272-275
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    • 1991
  • The effects of storage temperature and relative humidity (RH), and immersion treatment (30min) of sliced onions in various soluble starch solutions(1-3%; w/v) at $25^{\circ}C$ before air dehydration on caking and water absorption degree of powedered onion were analyzed. In the range of 15 to $35^{\circ}C$, the caking tendency of powdered onion increased with temperature at a constant RH (70% RH). The caking degree of powdered onion also increased as increasing the storage RH, at $25^{\circ}C$. Storage starchpreteatment significantly reduced the water absorption and caking degree of powdered onion under ambient condition. A pretreatment of soluble starch solution(3%) reduced almost 80% the caking of powdered onion compared to that of control.

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Furfural production from miscanthus and utilization of miscanthus residues (Miscanthus로부터 furfural 생산과 잔여물의 활용에 관한 연구)

  • Kim, Sung Bong;Yoo, Hah-Young;Lee, Sang Jun;Lee, Ja Hyun;Choi, Han Seok;Kim, Seung Wook
    • 한국신재생에너지학회:학술대회논문집
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    • 2011.11a
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    • pp.114.2-114.2
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    • 2011
  • Furfural is a versatile derivative. It can be utilized for a building-block of furfuryl alcohol production and a component of fuels or liquid alkanes. But in bio-process, furfural is a critical compound because it inhibits cell growth and metabolism. Furfural could be converted from xylose and usually produced from biomass in which hemicellulose is abundant. In this study, furfural production from miscanthus was performed and utilization of miscanthus residue was consequently conducted. At first, hydrolysis for investigation of miscanthus composition and furfural production was performed using sulfuric acid. Previously, we optimized dilute acid pretreatment condition for miscanthus pretreatment and the condition was found to be about 15 min of reaction time, 1.5% of acid concentration and about $140^{\circ}C$ of temperature and 60% (about 7 g/L) of xylose was solubilized from miscanthus. Using the xylose, furfural production was conducted as second step. Approximately $160{\sim}200^{\circ}C$ of temperature was accompanied with the hydrolysis for pyrolysis of biomass. When the investigated condition; $180^{\circ}C$ of temperature, 20 min of reaction time and 2% of acid concentration was operated for furfural production, furfural productivity was reached to be 77% of theoretical maximum. After reaction, residue of miscanthus was utilized as feedstock of ethanol fermentation. Residue was well washed using water and saccharified using hydrolysis enzymes. Hydrolysate (glucose) from saccharification was utilized for the carbon source of Saccharomyces cervisiae K35.

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Enhancing Enzymatic Saccharification of Corn Stover by Aqueous Ammonia Soaking Pretreatment (옥수수 줄기의 암모니아수 침지 전처리에 의한 효소 당화 향상)

  • Shin, Soo-Jeong;Yu, Ju-Hyun;Cho, Nam-Seok;Han, Sim-Hee;Kim, Mun-Sung;Park, Jong-Moon
    • Journal of the Korean Wood Science and Technology
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    • v.37 no.4
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    • pp.381-387
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    • 2009
  • Enhancing enzymatic saccharification of corn stover by aqueous ammonia soaking pretreatment was investigated on chemical compositional changes and enzymatic hydrolysis characteristics. At three different levels of aqueous ammonia soaking temperature and time ($140^{\circ}C$-1 h, $90^{\circ}C$-16 h and $50^{\circ}C$-6 days), higher temperature and shorter treatment time led to more xylan and lignin removal based on overall composition analysis and carbohydrate compositional analysis. More xylan and lignin removal in higher temperature treatment led to higher enzymatic saccharification of cellulose and xylan to monosaccharide by commercial cellulase mixtures (Celluclast 1.5L and Novozym 342 from Novozyme, Denmark).

Effects of Heat Pretreatment on the Quality under Storage of Fresh Ginger Rhizomes (저장 전 열처리가 생강의 이화학적 및 관능적 특성에 미치는 영향)

  • Chung, Hun-Sik;Lee, Hyeon-Jeong;Seong, Jong-Hwan;Moon, Kwang-Deog
    • Food Science and Preservation
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    • v.16 no.5
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    • pp.623-628
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    • 2009
  • Fresh ginger rhizomes were heat-treated for 10 min, 30 min, or 60 min using hot air ($40^{\circ}C$, $50^{\circ}C$, or $60^{\circ}C$), and stored in low-density polyethylene (0.04 mm thickness) bags for 2 months at $12{\pm}1^{\circ}C$. We studied the effects of heat pretreatment on changes in gas levels after packaging, and quality characteristics of the rhizomes. Oxygen and carbon dioxide levels progressively fell and rose, respectively, as the temperature of heat treatment rose and the duration of such treatment was extended. The sprouting rate of ginger rhizomes treated at $40^{\circ}C$ was higher than that of other samples. Rotting, softening, and increasing pH of rhizomes were accelerated by treatment at higher temperature for a longer time. Weight loss and soluble solid levels were not affected by heat treatment. Sensory qualities such as appearance, odor, and overall acceptability of rhizomes fell with treatment at a higher temperature for a longer time. These results suggest that heat pretreatment has a detrimental effect on the quality of fresh ginger rhizomes.

Evaluation of fiberization of rice hull by autohydrolysis conditons (자기가수분해 조건에 따른 농산부산물 왕겨의 섬유화 특성 평가)

  • Lee, Young-Ju;Jung, Woong-Ki;Sung, Yong-Joo
    • Korean Journal of Agricultural Science
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    • v.38 no.1
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    • pp.95-100
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    • 2011
  • In the present study, effects of autohydrolysis treatment on the properties of rice hull were investigated. Two temperature condition such as $160^{\circ}C$ and $180^{\circ}C$ and thee treatment time such as 15 min, 30 min and 45 min were applied for autohydrolysis treatment. The pH of extract after autohydrolysis treatment decreased as increasing of the temperature and the treatment time. The ash content and the lignin content of rice hull were not decreased by the autohydrolysis treatment. The structure of rice hull after autohydrolysis treatment became more weak, the better efficiency of the fiberization with PFI-Mill was shown for the autohydrolysis treated rice hull with the higher temperature and the longer treatment time.