• 한국수정란이식학회지
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• 제15권2호
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• pp.147-156
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• 2000

To investigate fatty acid constituents and relative compositions in the fluid of the follicles, oviducts, uterine body and uterine body in Korean native cow, the fluids of the reproductive tract were analyzed using gas chromatography. The samples were taken from various reproductive tract of 23 Korean native cows. q. Caprylic acid (C8:0), myristic acid(C14:0), palmitic acid(C16:0), palmitoleic acid(C16:1), stearic acid(C18:0), oleic acid(C18:1), linoleic acid(C18:2), arachidonic acid(C20:4) were found in the reproductive tracts of the cows, which made 8 kinds of fatty acid in total. 2. Palmitic acid, oleic acid, stearic acid were predominant with 35.67%, 24.98% and 17.52%, respectively. while low levels of fatty acids(<5%) were myristic acid, palmitoleic acid and caprylic acid with 1.75%, 1.28% and 2.69%, respectively. 3. Two kinds of polyunsaturated fatty acids, linoleic acid arachidonic acid were found in the reproductive tracts of cows. 4. Palmitic acid among saturated fatty acids and oleic acid among unsaturated fatty acids were the highest level in all of the reproductive tracts. 5. The Highest level of arachidonic acid was found in the uterine horn. 6. The sum of the palmitic acid and oleic acid were 61.72%, 63.72%, 57.66% and 57.65% for the fluid of follicle, oviduct, uterine horn and uterine body of the cows, respectively. 7. The relative compositions of palmitic acid, palmitoleic acid and caprylic acid were higher during the luteal phase than during the follicular phase. 8. The relative compositions of arachidonic acid was higher during the follicular phase in the fluid of uterine horn and uterine body of the cows. 9. The long chain fatty acid, the palmitic acid, stearic acid, oleic acid and linoleic acidshowed higher relative compositions during the follicular phase(86.49%∼95.51%) than during the luteal phase(85.64%∼88.93%).

• 한국가축번식학회지
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• 제26권4호
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• pp.311-320
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• 2002

The objective of this study was to assess the developmental potential in vitro produced embryos frozen-thawed with the various containers, and also examined expression and localization of heat shock protein 70 at these embryos. For the vitrification, 2-cell, 8-cell and blastocyst stage embryos produced by in vitro fertilization (IVF) and nuclear transfer (NT) were exposed the ethylene glycol 5.5 M freezing solution (EC 5.5) for 30 sec, loaded on each containers such EM grid, straw and cryo-loop, and then immediately plunged into liquid nitrogen. Thawed embryos were serially diluted in sucrose solution, each for 1 min. and cultured in CRI-aa medium. Survival rates of the vitrification production were assessed by re-expanded, hatched blastocysts. There were no differences in the survival rates of IVF using EM grid and cryo-loop. However, survival rates by straw were relatively lower than other containers. The use of cryo-loop resulted in only survival of nuclear transferred embryos (43.7%). Also, there embryos after IVF or NT were analysed by semi-quantitive reverse transcription-polymerase chain reaction (RT- PCR) methods for hsp 70 mRNA expression. Results revealed the expression of hsp 70 mRNh were higher thawed embryos than control embryos. Immunocytochemistry used to localize the hsp 70 protein in embryos. Two and 8-cell embryos derived under control condition was evenly distributed in the cytoplasm but appeared as aggregates in some frozen-thawed embryos. However, in the control, blastocysts displayed aggregate signal while Hsp70 in frozen-thawed blastocysts appeared to be more uniform In distribution. Therefore, this result suggests that the exploiting Hsp 70 in the early embryos may be role for protection of stress condition for increase viability of embryos within IVF, NT and there frozen-thawed embryos.

• 한국수정란이식학회지
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• 제23권1호
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• pp.25-30
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• 2008

This study was performed to identify and analyze the specifically expressed plasma proteins during early pregnancy in both pregnant and non-pregnant Hanwoo. Blood samples were collected at 0 (the day of AI), 2, 3, 4, 7, and 11 weeks after AI from pregnant (n=3) and non-pregnant (n=4) Hanwoo, respectively. The hematological parameters were measured. After 2-dimensional electrophoresis using serum, normalized protein spots were selected for the significant expression variation deviated over two fold in its expression level between two groups. Among 17 spots selected, 15 were identified as albumin, IgG1 heavy chain constant region, haptoglobin, ferrochelatase, fibrinogen, hemopexin. 5 spots were expressed only in non-pregnant specific. The spot identification of 1105 and 6106 was decreased after 3 weeks from AI. However, 2/17 spots were still unidentified. Further studies are needed to analyze the function of the proteins associated with early pregnancy.

• 공업화학
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• 제4권1호
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• pp.144-152
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• 1993

${\gamma}-Al_2O_3$, ${\alpha}-Al_2O_3$, $SiO_2$, $TiO_2$등에 고정화된 폴리에틸렌글리콜을 $Fe(CO)_5$에 의한 니트로벤젠의 상온 환원반응에 상이동 촉매로 사용하였다. 고정화된 PEG의 몰수는 담체의 비표면적에 따라 증가하였고 PEG/${\gamma}-Al_2O_3$가 가장 좋은 활성을 나타내었다. PEG의 사슬길이가 길고 NaOH 농도가 높을수록 반응속도가 증가하였다. 적외선분광기를 이용하여 반응기구에 대한 고찰도 함께 실시하였으며 폴리에틸렌글리콜 상이동 촉매는 본 반응의 활성물질로 알려진 $HFe(CO)_4{^{-}}$이온의 생성과 이의 유기상으로의 이동을 촉진시키는 것으로 판단되었다.

• 한국수정란이식학회지
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• 제25권4호
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• pp.281-286
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• 2010

This study was to evaluate the protein profile of seminal plasma using 2-DE in Hanwoo. Seminal plasma was harvested from five mature Hanwoo, and seminal plasma protein was extracted by M-PER Mammalian Protein Extraction Reagent. Proteins were refined by clean-up kit and quantified by Bradford method until total protein was $300\;{\mu}l$. Immobilized pH gradient (IPG) strip was used 18 cm and 3~11 NL. SDS-PAGE was used 12% acrylamide gel. Each gels were visualized by comassie brilliant blue and silver staining. These spots were analyzed by MALDI-TOF MS and searched on NCBInr. The result, 20 proteins of 36 protein spots were searched through peptide sequencing on the NCBInr. 8 proteins profiled by 2-DE were proved through previous bovine studies and the name of each protein was albumin, nucleobindin, clusterin, TIMP-2, spermadhesin Z13, spermadhesin-1 and BSP proteins (BSP 30 kDa and BSP A1/A2). 12 new proteins were ATP synthase, protein MAK16 homolog, Transmembrane protein 214, E3 ubiquitin-protein ligase BRE1A, dual serine/threonine and tyrosine protein kinase, tissue factor pathway inhibitor 2, alpha-actinin-4, RUN domain-containing protein 3B, catenin alpha-1, protein-glutamine gamma-glutamyltransferase 2, plakophilin-1 and inter-alpha-trypsin inhibitor heavy chain H1 has not been previously described in the bovine seminal plasma study. These proteins may be contribute to define the type of proteins affecting fertility of male and improve the fertilizing ability of semen in Hanwoo.

• 한국수정란이식학회지
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• 제24권2호
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• pp.77-87
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• 2009

To understand molecular and cellular mechanisms of many gene products in the female reproductive organs including the ovary and uterine endometrium as well as during embryo development, researchers have developed and utilized many effective methodologies to analyze gene expression in cells, tissues and animals over the last several decades. For example, blotting techniques have helped to understand molecular functions at DNA, RNA and protein levels, and the reverse transcription-polymerase chain reaction (RT-PCR) method has been widely used in gene expression analysis. However, some conventional methods are not sufficient to understand regulation and function of genes expressed in very complex patterns in many organs. Thus, it is required to adopt more high-throughput and reliable techniques. Here, we describe several techniques used widely recently to analyze gene expression, including annealing control based-PCR, differential display-PCR, expressed sequence tag, suppression subtractive hybridization and microarray techniques. Use of these techniques will help to analyze expression pattern of many genes from small scale to large scale and to compare expression patterns of genes in one sample to another. In this review, we described principles of these methodologies and summarized examples of comparative analysis of gene expression in female reproductive organs with help of those methodologies.

• 한국축산식품학회지
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• 제32권1호
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• pp.6-12
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• 2012

The association of three candidate genes, fatty acid synthase (FASN), microsomal triglyceride transfer protein (MTTP) and fatty acid binding protein 3 (FABP3), with fatty acid (FA) composition in Duroc pigs was investigated. Identified single nucleotide polymorphisms (SNPs) were used for polymerase chain reaction-restriction fragment length polymorphism genotyping. The c.265C>T SNP of FASN gene was significantly associated with high levels of palmitoleic acid (C16:1) (p<0.05), oleic acid (C18:1) (p<0.01), and mono-unsaturated fatty acid (MUFA) (p<0.01), but low levels of linoleic acid (C18:2) (p<0.01), alpha linolenic acid (C18:3) (p<0.05), and poly-unsaturated fatty acid (PUFA) (p<0.01) in animals having the CT genotype. The c.2573T>C SNP in the MTTP gene had a significant effect only in elevating the level of palmitoleic acid (C16:1) (p<0.05) in heterozygote animals. The polymorphism in FABP3 showed no significant effects on any fatty acid composition traits. These results suggest that the identified SNPs in the FASN and MTTP genes can be useful markers for selecting Duroc pigs having desirable healthy fatty acid composition.

• 대한화학회지
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• 제37권1호
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• pp.36-42
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• 1993

$Ca^{2+}$ 이온 존재하에서 비이온성 계면활성제, $nonylphenol-(ethylene oxide)_n\;[NP-(EO)_n; n = 11, 40, 100]$와 요오드간의 charge transfer (CT) 상호작용을 UV-Visible spectrophotometer를 이용하여 수용액중에서 조사하였다. Spectra의 특성은 $Ca^{2+}$ 이온 농도와 EO 갯수에 의존하였다. CMC 이상에서 $NP-(EO)_{11}$과 $NP-(EO)_{40}$에 대한 $Ca^{2+}$이온 첨가에 따른 CT band는 증가하다가 다시 감소하였으며, $NP-(EO)_{100}$에 대해서는 계속적인 증가를 보였다. CT band의 강도가 증가하는 것은 $Ca^{2+}$이온 존재하에서 미셀 구조가 더욱 조밀해진데 따른 요오드와의 donor-acceptor overlap 증가로 볼 수 있다. 이러한 현상들은 수용액 중에서 상태적으로 자유로이 여러가지 배향을 가질 수 잇는 선형 ethylene oxide(EO) 사슬이 유사 크라운 에테르 구조를 형성하여 $Ca^{2+}$ 이온과 착물을 형성할 수 있는 가능성을 보여준다.

• 한국수정란이식학회지
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• 제28권4호
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• pp.343-348
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• 2013

Cell-free fetal RNA has been highlighted as useful tools for the fetal sex determination or other genetic inherent disorder. However, there is no knowledge about the sex determination using cell free fetal RNA in bovine field. Thus, the present study aimed to evaluate the presence of transcripts of DDX3Y, USP9Y and ZRSR2Y genes in maternal plasma of pregnant cows to determine the sex of the fetus using real-time quantitative polymerase chain reaction assay, and verify its accuracy, sensitivity and specificity compared with the molecular testing and the calf sex at birth. Transcripts of USP9Y and DDX3Y genes were expressed in the all plasma of males and females both the control group and the experimental group. However, ZRSR2Y gene was matched up with the molecular testing and the true sex in control group and has an overall accuracy of 82.6%, a sensitivity of 75%, and a specificity of 100% in experimental group. Therefore, these results indicated that real time PCR technique, as a noninvasive and cost-efficient method, is possible to determination fetal sex in the bovine species using circulating cell free RNA in maternal plasma and especially ZRSR2Y gene could be a good candidate for the RNA based sex determination work.

• 한국수정란이식학회지
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• 제32권3호
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• pp.193-200
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• 2017

The purpose of this study is to confirm whether spontaneous adipocyte generation during chondrogenic induction culture affects the chondrogenic differentiation of porcine skin-derived stem cells (pSSCs). For this purpose, chondrogenic differentiation characteristics and specific marker gene expression were analyzed using cell lines showing different characteristics of spontaneous adipocyte formation. Of the four different lines of pSSCs, the pSSCs-IV line showed higher Oil red O (ORO) and glycosaminoglycan (GAG) extraction levels. Quantitative real-time polymerase chain reaction (qRT-PCR) analysis revealed that the levels of adipogenic markers peroxisome proliferator-activated receptor gamma 2 ($PPAR{\gamma}2$) and adipocyte Protein 2 (aP2) mRNAs were significantly higher in pSSCs-IV than those of the other pSSC lines (P<0.05). Among three chondrogenic markers, collagen type II (Col II) and sex determining region Y-box (Sox9) mRNAs were strongly expressed in pSSCs-IV (P<0.05), but not in aggrecan (Agg), which was significantly higher in pSSCs-II (P<0.05). These results demonstrate that the spontaneous adipocyte generation during chondrogenic differentiation has a positive effect on the chondrogenesis of pSSCs. More research is needed on the correlation between adipocyte generation and cartilage formation.