• Title/Summary/Keyword: Dilution protocol

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Dilution Reference Ranges by Predictive Value of Serum Level β-hCG in Early Pregnancy Viability (임신 초기 임신양상에 따른 혈청 β-hCG의 결과 예측에 의한 희석배수 참고치 설정)

  • Kim, Yoon Sik;Shin, Jang Yong;Seo, Yeong Mi;Yoo, Shin Soo
    • Korean Journal of Clinical Laboratory Science
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    • v.36 no.2
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    • pp.210-214
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    • 2004
  • This study was carried out to predict the value of serum ${\beta}$ subunit of humans chorionic gonadotropin(${\beta}$- hCG) in early pregnancy viability. This was performed among 85 women in vitro fertilization and embryo transfer(IVF-ET). The serum ${\beta}$-hCG levels were established for 30 normal singleton pregnancies, 10 twin and triplet pregnancies, 10 preclinical abortions, 10 clinical abortions, 20 biochemical abortions and 5 ectopic pregnancies. In comparison to normal singleton pregnancies, multiple pregnancies showed higher ${\beta}$-hCG. But clinical abortions, preclinical abortions and ectopic pregnancies showed lower ${\beta}$-hCG levels than singleton pregnancies. In conclusion, if we predict the value of serum ${\beta}$-hCG of variable early pregnancies and analyze it, we could predict the dilution protocol. Also, it can be useful in other ways.

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Fabrication and Application of Micro Polymer Chip Platform for Rare Cell Sample Preparation (희귀 세포 샘플 준비를 위한 마이크로 폴리머 칩 플랫폼 제작 및 활용)

  • Park, Taehyun
    • Journal of the Korea Convergence Society
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    • v.9 no.3
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    • pp.217-222
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    • 2018
  • In this paper, a new micro polymer chip platform and protocol were developed for rare cell sample preparation. The proposed platform and protocol overcome the current limitation of the dilution method which is based on statistics and the FACS method which expensive and requires fluorescence staining. It allows collecting exact number of target cells simply and selectively because the cells are visually confirmed during the collecting process. The collected cells can be transported or spiked into a desired locations, such as a microchamber, without cell loss. This research may applicable not only to a rare cell sample preparation for Lab on a Chip cancer diagnosis, but also to a single/double/multiple cell sample preparation for a cell analysis field. To verify this platform and protocol, five human breast cancer cells (MCF-7) were collected and transported into a hemocytometer chamber.

Optimization of Microwave-Assisted Method for Accelerated Glycated Hemoglobin Quantification from Amino Acids to Proteins

  • Tran, Thi Thanh Huong;Jeong, Ji-Seon
    • Mass Spectrometry Letters
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    • v.8 no.3
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    • pp.53-58
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    • 2017
  • Glycated hemoglobin ($HbA_{1c}$) has been commonly used to screen and diagnose for patients with diabetes mellitus. Here the accelerated procedure of microwave-assisted sample treatment from acid hydrolysis to enzyme digestion followed by isotope dilution liquid chromatography-tandem mass spectrometry (ID-LC-MS/MS) was optimized and applied to measure $HbA_{1c}$ in an effort to speed up analysis time. First, two signature peptides of $HbA_{1c}$ and hemoglobin $A_0$ were certified with amino acid analysis by setting optimized acid hydrolysis conditions to $150^{\circ}C$, 1.5 h and $10{\mu}M$ sample concentration in 8 M hydrochloric acid. Consequently, the accurate certified peptides above were used as calibration standards to implement the proteolytic procedure with endoproteinase Glu-C at $37^{\circ}C$, 700 W for 6 h. Compared to the traditional method, the microwave heating not only shortened dramatically sample preparation time, but also afforded comparable recovery yields. The optimized protocol and analytical conditions in this study are suitable for a primary reference method of $HbA_{1c}$ quantification with full SI-traceability and other similar proteins in complex biological samples.

Design and Implementation of Assisted GPS Navigation Systems Using TPEG Protocol of Terrestrial DMB Data Services (지상파 DMB 데이터 서비스의 TPEG프로토콜을 이용한 Assisted GPS 항법 시스템의 설계 및 구현)

  • Kim, Byung-Soo;Min, Seung-Wook
    • The Journal of Korean Institute of Communications and Information Sciences
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    • v.35 no.11B
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    • pp.1618-1623
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    • 2010
  • In this paper, we propose a new assisted global positioning system (A-GPS) using terrestrial digital multimedia broadcasting (T-DMB) data services. Because of the weak signal strength from GPS satellite and the signal blockage, it is difficult for the telematics terminal to determine the position in urban area. Proposed A-GPS system calculates pseudo range (PR) from timing information of GPS satellites and obtains the satellite information such as ephemeris from T-DMB station to determine the current position. Compared to conventional GPS system, the proposed system has better performance in terms of the fast time to first fix (TTFF), low horizontal dilution of precision (HDOP). Experimental results show that the proposed system is a feasible and robust solution.

A Preliminary Study for Development of a Bioassay Protocol Using the Sperm of a Starfish, Asterias amurensis

  • Ryu, Tae-Kwon;Lee, Chang-Hoon;Park, Jin-Woo
    • Proceedings of the Korea Society of Environmental Toocicology Conference
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    • 2003.10a
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    • pp.158-158
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    • 2003
  • Bioassays using gametes of sea urchins are widely used in ecotoxicological assessments of marine environments. Since most of sea urchin species in Korean coastal water spawn from spring to autumn, bioassay with them during the winter is impossible. In the course of developing standard methods for bioassays with Korean species, we found a winter-spawning starfish, Asterias amurensis, Since reproductive mode of asteroids is similar to echinoids, the bioassay protocol for sea urchins could be applied similarly to the starfish. Here, we tested and determined several conditions for the acceptability of bioassay with A. amurensis. The least required time for formation of fertilization membrane of fertilized eggs to be easily distinguished from unfertilized ones was 60 min. The threshold of sperm to egg ratio that could make acceptable fertilization rates in controls was 3000. The allowed time for manipulation of sperm after dilution in seawater was at most 3 hr. The optimal exposure time of sperms when the response against toxicant solution was relatively stable was in the range of 20-60 min. The tolerance range of sperms to the salinity of test solution was 26-38 psu. The sensitivity of A. amurensis sperm was intermediate among marine organisms commonly used in aquatic toxicity tests. The sperm bioassay with A. amurensis can be satisfactorily applied to toxicity assessments of marine environments.

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Aortic and Pulmograft Transplantation Utilizing Cryopreservation (초저온 냉동보관법을 이용한 동종판막 이식술에 대한 연구)

  • Song, Myeong-Geun;Lee, Dong-Sun
    • Journal of Chest Surgery
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    • v.23 no.4
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    • pp.622-639
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    • 1990
  • The use of aortic valve homograft has been developed since 1962 when Ross and Barratt - Boyes independently replaced a diseased aortic valve with an orthotopically inserted homograft valve. And also surgical treatment of complex congenital cardiac malformations utilizing homograft extracardiac conduit has been tried with better result than any other prosthetic material. The present study was undertaken to clarify the safety tissue viability, sterility, after following our protocol of procurement of heart, dissection of aortic and pulmonic homograft, sterilization, cryopreservation, thawing and dilution, and transplantation on experimental animal, sheep. Tissue viability of valve and great artery was assessed by tissue culture. Sterility was evaluated by bacterial and fungal culture. The method used was proven no deleterious effect on the integrity of the valve. Tissue culture of valve tissue before, and after cryopreservation process resulted that active fibroblast growth was observed from homograft sterilized with antibiotics. And culture of the transplanted homograft from sacrificed animal showed active fibroblast growth. Pathologic examination of implanted valve tissue from sacrificed sheep showed mild calcification and minor change, but there were moderate and severe calcification of wall of great arteries.

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A Study on Performance Evaluation of CEA kits (CEA 키트들 간의 성능 평가에 대한 고찰)

  • Ha, Dong hyuk;Sin, Hee Jung;You, Tae min;Noh, Kyoung Woon
    • The Korean Journal of Nuclear Medicine Technology
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    • v.22 no.1
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    • pp.76-79
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    • 2018
  • Purpose Carcinoembryonic antigen (CEA) is cell-surface 180-200 kDa glycoprotein that is overexpressed in breast, stomach pancreas, lung, and colorectal cancers. CEA was first described in 1965 by Gold and Freedman and then serum CEA of colorectal cancer patients was first measured in 1969 by radioimmunoassay by Thomson. CEA is currently most widely used tumor marker in the clinic for management of colorectal cancer. Various CEA test kits have been developed and commercialized. CEA kits from different manufacturers might have different test results because of different reagents and protocol. The purpose of this study was to compare results of four commercial available CEA kits. Materials and Methods This study was designed to evaluate four commercially available CEA kits using serum samples acquired from 120 patients who visited our clinic. Test results were compared and analyzed according to the respective test methods. High concentration samples were diluted with saline and diluted solution. Results All of the four kits showed a significant correlation within the reference value. However, three of the four kits used for the dilution test using high concentration samples showed the hook effect. Conclusion Results of the present study showed that It is important to establish the standardized dilution standards for the high-concentration specimens to manage the error of the test result by the hook effect.

A Comparative Study on Fresh and Frozen Embryo Transfer after Superovulation in Black Bengal Goats(Capra-hircus)

  • Mishra, O.P.;Pandey, J.N.;Gawande, P.G.
    • Journal of Embryo Transfer
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    • v.17 no.2
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    • pp.91-100
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    • 2002
  • The experiment was divided into two phases. In phase-I fresh embryos were transferred and in Phase-II frozen embryos were transferred. Embryos were collected by using Dulbecco's phosphate buffered saline. In phase-I total of 65 ova were collected out of 107 ovulation in 18 goats. Recovery of ova was 60.74%, of which 51 (78.46%) was fertilized. Sixteen embryos were transferred to 10 recipient goats and kidding was observed in 6 goats, that produced 10 kids. Thus, 62.50% embryo survival and 60% kidding were achieved in phase-I. In phase-II of the experiment, 17 regular cyclic Black Bengal goats were used. The main purpose was to study the viability of caprine embryos after cryopreservation. In this phase the embryos were collected and frozen using Bio-cool freezers. A two step addition of cryoprotectants (5% glycerol and 10% glycerol) and three-step dilution of cryoprotectants with 1mole (M) sucrose was used. Embryos were preserved for 10 to 45 days. Out of 27 embryos preserved, 18 were recovered after freezing and thawing (37$^{\circ}C$ water bath) with 33.33% embryonic loss. Seventeen frozen and thawed embryos were transferred in 9 recipient goats, out of which kidding was observed in 6 goats and 7 kids were produced, giving a 66.66% kidding and embryo survival of 41.17%. The technique utilized for fresh and frozen embryo transfer can be successfully utilized to produce goats of superior genetic merits. The protocol used for addition of cryoprotectant, freezing, thawing and dilution was found suitable for caprine embryo freezing.

Analysis for Concentration Range of Fluorescein Sodium (플루오레신나트륨의 농도 범위 분석)

  • Lee, Da-Ae;Kim, Yong-Jae;Yoon, Ki-Cheol;Kim, Kwang-Gi
    • Journal of Biomedical Engineering Research
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    • v.41 no.2
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    • pp.67-74
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    • 2020
  • Brain tumors or gliomas are fatal cancer species with high recurrence rates due to their strong invasiveness. Therefore, the goal of surgery is complete tumor resection. However, the surgery is difficult to distinguish the border because tumors and blood vessels have the same color tone and shape. The fluorescein sodium is used as a fluorescence contrast agent for boundary separation. When the external light source is irradiated, yellow fluorescence is expressed in the tumor, which helps distinguish between blood vessels and tumor boundaries. But, the fluorescence expression of fluorescence sodium depends on the concentration of fluorescein sodium and such analytical data is insufficient. The unclear fluorescence can obscure the boundaries between blood vessels and tumors. In addition, reduce the efficiency of fluorescence sodium use. This paper proposes a protocol of concentration range for fluorescence expression conditions. Fluorescent expression was observed using a near-infrared (NIR) color camera with corresponding dilution using normal saline in 1 ml microtube. The flunoresence emission density range is 1.00 mM to 0.15 mM. The fluorescence emission begin to 1.00 mM and the 0.15 mM discolor. The discolor is difficult to fluorescence emission condition obserbation. Thus, the maximum density range of the bright fluoresecein is 0.15 mM to 0.30 mM. When the concentration range of fluorescein sodium is analyzed based on the gradient of fluorescence expression and the power measurement, the brightest fluorescence is expected to facilitate the complete resection of the tumor. For the concentration range protocol, setting concentration ranges and analyzing fluorescence expression image according to saturation and brightness to find optimal fluorescence concentration are important. Concentration range protocols for fluorescence expression conditions can be used to find optimal concentrations of substances whose expression pattern varies with concentration ranges. This study is expected to be helpful in the boundary classification and resection of brain tumors and glioma.

Development of Protocol for the Effective Detection of Feline Calicivirus as Norovirus Surrogate in Oyster and Lettuce (굴과 상추에서 노로바이러스의 대체모델 feline calicivirus의 효율적 검출법 개발)

  • Lee, Soo-Yeon;Jang, Keum-Il;Woo, Gun-Jo;Kwak, Hyo-Sun;Kim, Kwang-Yup
    • Korean Journal of Food Science and Technology
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    • v.39 no.1
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    • pp.71-76
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    • 2007
  • Foodborne illness caused by Noroviruses (NVs) is increasing rapidly in Korea. This study developed an effective detection protocol for NVs found in contaminated oysters and lettuce through an investigation using the major steps of virus particle separation, concentration and RT-PCR. As a surrogate model for NVs, the cultivable feline calicivirus (FCV) that belongs to the same Caliciviridae family was used. Instead of using a time-consuming ultracentrifugation method, efficient methods based on solvent extraction and PEG precipitation procedure were applied. Direct homogenization of a 25g sample of whole oyster and lettuce in 175mL PBS provided the simplicity that would be needed in the actual field of food product examination. The overnight PEG precipitation step at $4^{\circ}C$ was reduced to 3 h by placing the reaction tube in ice and by adjusting the PEG concentrations. The application of the use of chloroform and 0.2 ${\mu}m$ syringe filtration together showed a better detection efficiency than the use of chloroform alone in removing PCR inhibitors for both oyster and lettuce samples. Also, dilution of the extracted RNA solution before PCR provided increased sensitivity. The improved detection protocol developed in this study could be efficiently applied to detect FCV and most likely NVs from oysters and lettuce.