• Title/Summary/Keyword: Liquid nitrogen

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EVALUATION OF THE VIABILITY OF PERIODONTAL LIGAMENT CELL IN RAT TEETH USING SLOW CRYOPRESERVATION METHOD WITH MAGNETIC FIELD (자기장 저속 냉동보관법을 이용한 쥐 치아 치주인대세포의 활성도 검사)

  • Ahn, Hyun-Jung;Kim, Eui-Seong;Kim, Jin;Kim, Duck-Won;Kim, Ki-Yeol;Lee, Chan-Young;Lee, Seung-Jong
    • Restorative Dentistry and Endodontics
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    • v.33 no.4
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    • pp.332-340
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    • 2008
  • The purpose of this study was to evaluate the viability of periodontal ligament cell in rat teeth using slow cryopreservation method with magnetic field through MTT assay and TUNEL test. For each group, 12 teeth of 4 weeks old white female Sprague-Dawley rat were used for MTT assay, and 6 teeth in TUNEL test. The Maxillary left and right, first and second molars were extracted as atraumatically as possible under tiletamine anesthesia. The experimental groups were group1 (immediately extraction), group 2 (cold preservation at 4$^{\circ}C$ for 1 week), group 3 (rapid cryopreservation in liquid nitrogen), group 4 (slow cryopreservation with magnetic field of 1 G), and group 5 (slow cryopreservation). F medium was used as preservation medium and 10% DMSO as cryoprotectant. After preservation and thawing, the MTT assay and TUNEL test were processed. One way ANOVA and Scheffe method were performed at the 95% level of confidence. The value of optical density obtained after MTT analysis was divided by the value of eosin staining for tissue volume standardization. In both MTT assay and TUNEL test, it had showed no significant difference among group 3, 4, and 5. And group 3 had showed higher viability of periodontal ligament cell than group 2. From this study, slow cryopreservation method with magnetic field can be used as one of cryopreservation methods.

Influences of Rate of Artificial Insemination Following Estrus Induction in Dog (개에서 발정유도가 인공수정효율에 미치는 영향)

  • 이영락;강태영;최상용
    • Journal of Embryo Transfer
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    • v.18 no.1
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    • pp.61-68
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    • 2003
  • Considerable attention has been focused on the cryopreservation of semen and estrus induction in dog, as consequence of poor productivity caused by long anestrus period, in order to enhance the productivity of youngs and to preserve the breeds. The objectives of this study were to improve reproductive efficiency of artificial insemination with fresh- and frozen-semen following estrus induction in dog. Fifty infertilie dogs (age 2~3 years) were selected fur the study and divided into three different estrus induction treatment groups. Group 1 : dogs (n=15) were given clomifene (0.1 mg/kg) orally f3r five days at 12 hr intervals. Croup 2: dogs (n=15) were given bromocriptine (50 $\mu$g/kg) orally for five days at 12 hr intervals, followed by single injection intravenously of 500 IU GnRH (Croup 3, n=20) when pro-estrus occurred. After being treated, the dogs were evaluated fur the rates of estrus induction and time interval lapses from treatment to beginning of the pro-estrus. The rates of pregnancy in estrus inducted dogs mated naturally compared to those inseminated artificially with ejaculated fresh semen and frozen-thawed semen. Estrus detection was performed using the method of vaginal smear and confirmed by the plasma progesterone assay. Pregnancy was confirmed by ultrasonograpy on day 25, 35 and 55 post insemination. The ejaculated semen was exposed to a mixture of Tris extender with cryoprotectant (Trisma, 81 mM; TES, 209 mM; citric acid, 6 mM; glucose, 5 mM; glycerol, 8%) and cryopreserved gradually by slow-cooling at 17 co above the surface of liquid nitrogen (L$N_2$) for 23 min. The use of fresh semen, the pregnancy rates were observed 66.6, 66.6, 75.0 and 83.3% in natural estrus, clomifene induced, bromocriptine induced and a combination of GnRH and bromocriptine, respectively. The use of frozen-thawed semen, the pregnancy rates were observed 66.6, 33.3, 50.0 and 60.0% in natural estrus, clomifene induced, bromocriptine induced and a combination of GnRH and bromocriptine, respectively. No difference was observed in the number of offspring produced among natural estrus and treated groups inseminated with fresh or frozen-thawed semen. In conclusion, there was no significant differences in the pregnancy rate of dogs between group treated with a combination of GnRH and bromocriptine and group treated clomifene or bromocriptine only. However, frozen-thawed semen can be used successfully fur artificial insemination in dog.

Modern Paper Quality Control

  • Olavi Komppa
    • Proceedings of the Korea Technical Association of the Pulp and Paper Industry Conference
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    • 2000.06a
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    • pp.16-23
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    • 2000
  • The increasing functional needs of top-quality printing papers and packaging paperboards, and especially the rapid developments in electronic printing processes and various computer printers during past few years, set new targets and requirements for modern paper quality. Most of these paper grades of today have relatively high filler content, are moderately or heavily calendered , and have many coating layers for the best appearance and performance. In practice, this means that many of the traditional quality assurance methods, mostly designed to measure papers made of pure. native pulp only, can not reliably (or at all) be used to analyze or rank the quality of modern papers. Hence, introduction of new measurement techniques is necessary to assure and further develop the paper quality today and in the future. Paper formation , i.e. small scale (millimeter scale) variation of basis weight, is the most important quality parameter of paper-making due to its influence on practically all the other quality properties of paper. The ideal paper would be completely uniform so that the basis weight of each small point (area) measured would be the same. In practice, of course, this is not possible because there always exists relatively large local variations in paper. However, these small scale basis weight variations are the major reason for many other quality problems, including calender blacking uneven coating result, uneven printing result, etc. The traditionally used visual inspection or optical measurement of the paper does not give us a reliable understanding of the material variations in the paper because in modern paper making process the optical behavior of paper is strongly affected by using e.g. fillers, dye or coating colors. Futhermore, the opacity (optical density) of the paper is changed at different process stages like wet pressing and calendering. The greatest advantage of using beta transmission method to measure paper formation is that it can be very reliably calibrated to measure true basis weight variation of all kinds of paper and board, independently on sample basis weight or paper grade. This gives us the possibility to measure, compare and judge papers made of different raw materials, different color, or even to measure heavily calendered, coated or printed papers. Scientific research of paper physics has shown that the orientation of the top layer (paper surface) fibers of the sheet paly the key role in paper curling and cockling , causing the typical practical problems (paper jam) with modern fax and copy machines, electronic printing , etc. On the other hand, the fiber orientation at the surface and middle layer of the sheet controls the bending stiffness of paperboard . Therefore, a reliable measurement of paper surface fiber orientation gives us a magnificent tool to investigate and predict paper curling and coclking tendency, and provides the necessary information to finetune, the manufacturing process for optimum quality. many papers, especially heavily calendered and coated grades, do resist liquid and gas penetration very much, bing beyond the measurement range of the traditional instruments or resulting invonveniently long measuring time per sample . The increased surface hardness and use of filler minerals and mechanical pulp make a reliable, nonleaking sample contact to the measurement head a challenge of its own. Paper surface coating causes, as expected, a layer which has completely different permeability characteristics compared to the other layer of the sheet. The latest developments in sensor technologies have made it possible to reliably measure gas flow in well controlled conditions, allowing us to investigate the gas penetration of open structures, such as cigarette paper, tissue or sack paper, and in the low permeability range analyze even fully greaseproof papers, silicon papers, heavily coated papers and boards or even detect defects in barrier coatings ! Even nitrogen or helium may be used as the gas, giving us completely new possibilities to rank the products or to find correlation to critical process or converting parameters. All the modern paper machines include many on-line measuring instruments which are used to give the necessary information for automatic process control systems. hence, the reliability of this information obtained from different sensors is vital for good optimizing and process stability. If any of these on-line sensors do not operate perfectly ass planned (having even small measurement error or malfunction ), the process control will set the machine to operate away from the optimum , resulting loss of profit or eventual problems in quality or runnability. To assure optimum operation of the paper machines, a novel quality assurance policy for the on-line measurements has been developed, including control procedures utilizing traceable, accredited standards for the best reliability and performance.

Isolation and Morphological Identification of Fresh Water Green Algae from Organic Farming Habitats in Korea (유기농업 생태계로부터 담수 녹조류 분리 및 형태적 동정)

  • Kim, Min-Jeong;Shim, Chang-Ki;Kim, Yong-Ki;Hong, Sung-Jun;Park, Jong-Ho;Han, Eun-Jung;Jee, Hyeong-Jin;Yun, Jong-Chul;Kim, Suk-Chul
    • Korean Journal of Organic Agriculture
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    • v.22 no.4
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    • pp.743-760
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    • 2014
  • This study aimed to isolate and identify freshwater algae from the organic agricultural ecosystems and investigate its biological characteristics to study the possibility of utilizing a biomass freshwater algae in organic farming. In the survey area, average water temperature was $12.4{\sim}28.2^{\circ}C$ and the pH ranges were from 6.1 to 8.5. The solid culture method is more suitable than liquid culture method for isolation of freshwater algae with lower contamination level and higher isolation frequency. A total of 115 strains were isolated from six freshwater algae habitats in nine regions in Korea. BGMM (BG11 Modified Medium) amended with NaNO3 and $KNO_3$ as a nitrogen, and $Na_2CO_3$ as carbon source was designed to isolate and culture freshwater algae. Absorbance of freshwater algae culture has increased dramatically to four days and decreased after eight days after inoculation. CHK008 of the seven isolates showed the highest absorbance in seven days after culturing in BGMM. The optimal pH of BGMM for culturing freshwater algae was pH 6-7. As light intensity increased, growth of freshwater algae increased. Among the five kinds of carbon sources, glucose and galactose promoted good growth of freshwater algae in BGMM. The colony color of purified 16 green algae isolates showed a separation of green, dark and light green, and of them, eleven algae strains showed a strong fluorescent light under fluorescence microscopy. Cell size of the green algae showed a wide range of variation depending on the species. General morphology of the green algae strains was spherical. Chlamydomonas sp. was elliptical, and Chlorella sorokiniana was ellipsoidal and cylindrical. All strains of the green algae except for Chlamydomonas sp. did not have flagella. One isolate of Chlamydomonas sp. and five isolates of C. sorokiniana secreted mucus. Sixteen isolates of 16 green algae were identified as two family and six species, Chlorella vulgalis, C. sorokiniana, C. pyrenoidosa, C. kessleri, C. emersonii, and Chlamydomonas sp. based on their morphological characteristics.

EVALUATION OF PERIODONTAL LIGAMENT CELL VIABILITY IN RAT TEETH AFTER FROZEN PRESERVATION USING IN-VIVO MTT ASSAY (급속냉동된 쥐 치아의 in vivo MTT 검색법을 이용한 치주인대세포 활성도 평가)

  • Kim, Jae-Wook;Kim, Eui-Sung;Kim, Jin;Lee, Seung-Jong
    • Restorative Dentistry and Endodontics
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    • v.31 no.3
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    • pp.192-202
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    • 2006
  • The purpose of this study was to examine the viability of PDL cells in rat molars by using in vivo MTT assay, which was used to compare fast cryopreservation group by liquid nitrogen $(-196^{\circ}C)\;with\;4^{\circ}C$ cold preservation group. A total of 74 Sprague-Dawley white female rats of 4 week-old with a body weight of 100 grams were used. The maxillary left and right, first and second molars were extracted as atraumatically as possible under ketamine anesthesia. Ten teeth of each group were divided as six experimental groups depending upon the preservation. Cryopreservation groups were Group 1 (5% DMSO 6% HES in F medium) Group 2 (10% DMSO in F medium), Group 3 (5% DMSO 6% HES in $Viaspan^(R)$). Group 4 (10% DMSO in $Viaspan^(R)$) which were cryopreserved for 1 week and cold preservation groups were Group 5 (F medium) , Group 6 ($Viaspan^(R)$) at $4^{\circ}C$ for 1 week. Immediate extraction group was used as a control. After preservation and thawing, the in vivo MTT assay was processed. Two way ANOVA and Duncan's Multiple Range Test was performed at the 95 % level of confidence, Another 2 teeth of each group were treated as the same manner and frozen sections $10{\mu}m$ thick for microscopic observation. The value of optical density obtained after in vivo MTT analysis was divided by the value of eosin staining for tissue volume standardization. Group 1, 2 had significantly higher optical density than Group 3 and 4 which had the lowest OD value. Group 6 had higher OD value than in Group 5 (P<0.05). Histological findings of periodontal ligament cell, after being stained with MTT solution were consistent with the in vivo MTT assay results. In this study, the groups which were frozen with DMSO as a cryoprotectant and the groups with F medium showed the best results.

Cryopreservation of Umbilical Cord as a Source of Mesenchymal Stromal Cells and Growth Factors (간엽줄기세포와 성장인자의 공급원으로서 제대 조직의 동결 보관)

  • Lee, Hye Ryun;Roh, Eun Youn;Shin, Sue;Yoon, Jong Hyun;Kim, Byoung Jae;Jeon, Hye Won
    • The Korean Journal of Blood Transfusion
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    • v.23 no.2
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    • pp.115-126
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    • 2012
  • Background: Umbilical cord (UC) is a promising source of mesenchymal stromal cells (MSCs). We compared the characteristics of MSCs from cryopreserved UC with those from fresh tissues, and demonstrated the possibility of UC cryopreservation for acquisition of MSCs from cryopreserved UC. Methods: Each UC was sliced into two types ($1{\sim}2mm^3$ vs. 0.5 cm), and cryopreserved in liquid nitrogen using different media (autologous cord blood plasma, aCBP vs. RPMI 1640). A fresh aliquot of $1{\sim}2mm^3$-sized UC was used as control tissue. After one week, the cryopreserved tissues were thawed and cultured. For the 0.5 cm UC, a slicing step into $1{\sim}2mm^3$ was needed. Cell count, viability, proliferative activity, and surface antigens were determined from harvested MSCs. Several growth factors (EGF, IGF-1, PDGF, TGF-${\beta}$, bFGF, and VEGF), were measured from the culture supernatant. Results: Eleven UC were enrolled in the study. Efficiencies of obtaining MSCs were higher in cryopreserved UC using RPMI 1640, compared with use of aCBP; the same result was observed for 0.5 cm sized UC, compared with $1{\sim}2mm^3$ sized UC. No difference in proliferative activity was observed between MSCs from fresh and cryopreserved UC. The amount of growth factors in culture supernatant using RPMI 1640 was larger than that of fresh tissues. Conclusion: We obtained growth factors from the supernatant as well as MSCs from cryopreserved UC. As with a cord blood bank, in the future, cryopreservation of UC for acquisition of both MSCs and growth factors would be possible in a time of need.

Investigation of ground condition charges due to cryogenic conditions in an underground LNG storage plant (지하 LNG 저장 시험장에서 극저온 환경에 의한 지반상태 변화의 규명)

  • Yi Myeong-Jong;Kim Jung-Ho;Park Sam-Gyu;Son Jeong-Sul
    • Geophysics and Geophysical Exploration
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    • v.8 no.1
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    • pp.67-72
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    • 2005
  • To investigate the feasibility of a new concept of storing Liquefied Natural Gas (LNG) in a lined hard rock cavern, and to develop essential technologies for constructing underground LNG storage facilities, a small pilot plant storing liquid nitrogen (LN2) has been constructed at the Korea Institute of Geoscience and Mineral Resources (KIGAM). The LN2 stored in the cavern will subject the host rock around the cavern to very low temperatures, which is expected to cause the development of an ice ring and the change of ground condition around the storage cavern. To investigate and monitor changes in ground conditions at this pilot plant site, geophysical, hydrogeological, and rock mechanical investigations were carried out. In particular, geophysical methods including borehole radar and three-dimensional (3D) resistivity surveys were used to identify and monitor the development of an ice ring, and other possible changes in ground conditions resulting from the very low temperature of LN2 in the storage tank. We acquired 3D resistivity data before and after storing the LN2, and the results were compared. From the 3D images obtained during the three phases of the resistivity monitoring survey, we delineated zones of distinct resistivity changes that are closely related to the storage of LN2. In these results, we observed a decrease in resistivity at the eastern part of the storage cavern. Comparing the hydrogeological data and Joint patterns around the storage cavern, we interpret this change in resistivity to result from changes in the groundwater flow pattern. Freezing of the host rock by the very low temperature of LN2 causes a drastic change in the hydrogeological conditions and groundwater flow patterns in this pilot plant.

Improvement of an Analytical Method for Methoprene in Livestock Products using LC-MS/MS (LC-MS/MS를 이용한 축산물 중 살충제 메토프렌의 잔류분석법 개선)

  • Park, Eun-Ji;Kim, Nam Young;Park, So-Ra;Lee, Jung Mi;Jung, Yong Hyun;Yoon, Hae Jung
    • Journal of Food Hygiene and Safety
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    • v.37 no.3
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    • pp.136-142
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    • 2022
  • The research aims to develop a rapid and easy analytical method for methoprene using liquid chromatography-tandem mass spectrometry (LC-MS/MS). A simple, highly sensitive, and specific analytical method for the determination of methoprene in livestock products (beef, pork, chicken, milk, eggs, and fat) was developed. Methoprene was effectively extracted with 1% acetic acid in acetonitrile and acetone (1:1), followed by the addition of anhydrous magnesium sulfate (MgSO4) and anhydrous sodium acetate. Subsequently, the lipids in the livestock sample were extracted by freezing them at -20℃. The extracts were cleaned using MgSO4, primary secondary amine (PSA), and octadecyl (C18), which were then centrifuged to separate the supernatant. Nitrogen gas was used to evaporate the supernatant, which was then dissolved in methanol. The matrix-matched calibration curves were constructed using 8 levels (1, 2.5, 5, 10, 25, 50, 100, 150 ng/mL) and the coefficient of determination (R2) was above 0.9964. Average recoveries spiked at three levels (0.01, 0.1, and 0.5 mg/kg), and ranged from 79.5-105.1%, with relative standard deviations (RSDs) smaller than 14.2%, as required by the Codex guideline (CODEX CAC/GL 40). This study could be useful for residue safety management in livestock products.

Improvement of Oxygen Isotope Analysis in Seawater samples with Stable Isotope Mass Spectrometer (질량분석기를 이용한 해수 중 산소안정동위원소 분석법의 개선)

  • Park, Mi-Kyung;Kang, Dong-Jin;Kim, Kyung-Ryul
    • The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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    • v.13 no.4
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    • pp.348-353
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    • 2008
  • Oxygen isotope has not been used actively in water mass studies because of difficulties on the analysis though it has advantages as a water mass tracer. The most popular method to analysis the oxygen isotope ratio in water samples is equilibration method: isotopic equilibrium of water with $CO_2$ at constant temperature. The precision of oxygen isotope analysis using commercial automatic $H_2O/CO_2$ equilibrator is ${\pm}0.1%o$. This value is not sufficient for studies in open ocean. The object of this study is to improve the analytical precision enough to apply open ocean studies by modification of the instrument. When sample gas is transferred by the pressure difference, the fractionation which is preferential transportation of light isotope can be occurred since the long transportation path between the equilibrator and mass spectrometer. And the The biggest source of error during the analysis is long distance and large volume of the pathway of sample gas between. Therefore, liquid nitrogen trap and high vacuum system are introduced to the system. The precisions of 14 time analysis of same seawater sample are ${\pm}0.081%o$ and ${\pm}0.021%o$ by built-in system and by modified system in this study, respectively.

Effects of Co-digestate application on the Soil Properties, Leachate and Growth Responses of Paddy Rice (통합혐기소화액의 시용이 벼 생육 및 논토양 환경에 미치는 영향)

  • Hong, Seung-Gil;Shin, Joung-Du;Kwon, Soon-Ik;Park, Woo-Kyun;Lee, Deog-Bae;Kim, Jeong-Gyu
    • Journal of the Korea Organic Resources Recycling Association
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    • v.18 no.4
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    • pp.31-37
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    • 2010
  • Livestock manures have a potential to be a valuable resource with an efficient treatment. In Korea, 42 million tons of livestock manure were generated in 2008, and 84 % of them were used for compost and liquid fertilizer production. Recently recycling of livestock manure for biogas production through anaerobic digestion is increasing, but its utilization in agriculture is still uncertified. In this study, there was applied co-digestate to the paddy for rice cultivation based on N supplement. Co-digestate was fertilizer fermented with pig slurry and food waste combined with the ratio of 70:30(v:v) in its volumetric basis. For assessing the safety of co-digestate, it was monitored the contents of co-digestate for seasonal variation, resulted in no potential harm to the soil and plant by heavy metals. The results showed that soil applied with co-digestate was increased in exchangeable potassium, copper and zinc mainly due to the high rate of pig slurry in co-digestate applied. Considering high salt content due to the combination with food waste, strict quality assurances are needed for safe application to arable land though it has valuable fertilizer nutrient. Leachate after treatment showed that the concentration of nitrate nitrogen washed out within two weeks. Considering the salt accumulation results in soil, it is highly recommended that the application rate of co-digestate should not exceed the crop fertilization rate based on N supplement. With these results, it was concluded that co-digestate could be used as an alternative fertilizer for chemical fertilizer. More study is needed for the long-term effects of co-digestate application on the soil and water environment.