• Asian-Australasian Journal of Animal Sciences
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• 제31권4호
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• pp.489-498
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• 2018

Objective: Foot and mouth disease (FMD) and porcine reproductive and respiratory syndrome (PRRS) are major diseases that interrupt porcine production. Because they are viral diseases, vaccinations are of only limited effectiveness in preventing outbreaks. To establish an alternative multi-resistant strategy against FMD virus (FMDV) and PRRS virus (PRRSV), the present study introduced two genetic modification techniques to porcine cells. Methods: First, cluster of differentiation 163 (CD163), the PRRSV viral receptor, was edited with the clustered regularly interspaced short palindromic repeats-CRISPR-associated protein 9 technique. The CD163 gene sequences of edited cells and control cells differed. Second, short hairpin RNA (shRNAs) were integrated into the cells. The shRNAs, targeting the 3D gene of FMDV and the open reading frame 7 (ORF7) gene of PRRSV, were transferred into fibroblasts. We also developed an in vitro shRNA verification method with a target gene expression vector. Results: shRNA activity was confirmed in vitro with vectors that expressed the 3D and ORF7 genes in the cells. Cells containing shRNAs showed lower transcript levels than cells with only the expression vectors. The shRNAs were integrated into CD163-edited cells to combine the two techniques, and the viral genes were suppressed in these cells. Conclusion: We established a multi-resistant strategy against viral diseases and an in vitro shRNA verification method.

• Journal of Microbiology and Biotechnology
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• 제10권5호
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• pp.707-713
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• 2000

Myasthenia gravis (MG) is caused mainly by autoantibodies directed against acetylcholine receptors located in the postsynaptic muscle cell membrane. Using in vitro selection techniques, we isolated an RNA containing 2'-fluoro pyrimidines that can specifically and avidly ($K_d$ ~25 nM) bind rat monoclonal antibody called mAb198, which recognizes the main immunogenic region on the acetylcholine receptors. This RNA can act as a very effective decoy and block mAb198 binding to the receptors in vitro. Furthermore, this RNA decoy can prevent the antigenic modulation of the acetylcholine receptor caused by mAb198 in human muscle cell cultures with and $IC_{50} $of approximately $2.4{\mu}M$. These results indicate that the RNA selected in this study is a more potent decly inhibitor of myashthenic antibodies than the previously identified RNA with 2'-amino pyrimidines [11]. Moreover, this RNA cross-reacts with autoantibodies from patients with MG and can protect human cells from the effects of these antibodies. These observations have important implications for developing an antigen-specific treatment of autoimmune diseases including MG, which is based on decoy RNAs selected in vitro.

• 한국수정란이식학회지
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• 제15권1호
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• pp.95-102
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• 2000

Techniques for manipulation of spermatozoa and oocytes have been widely used for in vitro production(IVP) of Hanwoo. This study was conducted to examine the effects of theophylline and heparin on frozen-thawed Hanwoo sperm for enhancing the efficiency of IVP technique. Oocytes were inseminated with forzen bull semen treated with either theophylline or heparin for examining the effect of each substance on fertilization and subsequent development. More (P<0.05) oocytes formed pronucleus and develop to the morula and blastocyst stages after inseminated with sperm treated with heparin than after inseminated with sperm treated with theophylline. The pregnancy rate after embryo transfer was higher after heparin treatment than after theophylline treatment, but did not differ significantly. There was no significant difference of offspring delivery between two groups. In conculsion, theophylline and heparin can be used for enhancing the efficiency of IVP system for Hanwoo. Considering characteristics of these substance, theophylline may be useful in the artificial insemination system, which requires vigorous sperm motility. While, heparin supporting sperm viability in vitro can be effectively used for improving in vitro-fertilization system.

• 한국동물번식학회:학술대회논문집
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• 한국동물번식학회 2001년도 발생공학 국제심포지움 및 학술대회 발표자료집
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• pp.48-49
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• 2001

While transgenic manipulation in mice have been very successful the same is not true for cattle and pigs. The inability to isolate ES cells from the bovine and porcine has precluded the utilization of the gene targeting technology in these species. Fortunately new advances in cloning by nuclear transfer have opened up a unique opportunity to undertake precise genetic modification in cattle and pigs. The ability of a number of different laboratory groups to successfully clone cattle is due to numerous research programs focused on nuclear transfer in cattle, and the enormous base of knowledge developed over the last 20 years involving the application of assisted reproductive techniques in cattle. Successful and repeatable procedures for in vitro oocyte maturation, in vitro fertilization, and in vitro embryo culture are now well established for cattle. In our laboratory we have utilized nuclear transfer to reproduce the genotypes of several animals, selected for cloning based on their inherent genetic value. Results that we have obtained to date are similar to those reported by other laboratories. (omitted)

• 전자통신동향분석
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• 제37권3호
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• pp.33-40
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• 2022

Communication technologies are evolving to meet the ever-increasing demand for fast transmission speeds and large capacity. However, the introduction of new mobile networks, such as 5G, should be accompanied by changes in corresponding frequency and modulation techniques to enable the commercialization of its services. Hence, investigating the human health effects of electromagnetic exposures used in these new communication systems is crucial. Since in vitro electromagnetic experiments can provide essential information on important mechanisms of exposure effects, this article presents in vitro exposure systems necessary for reliable experiments. Moreover, the fundamental requirements for system implementation and the features of various systems are discussed.

• Asian-Australasian Journal of Animal Sciences
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• 제29권8호
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• pp.1136-1144
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• 2016

Leucaena silage was supplemented with different levels of molasses and urea to study its nutritive value and in vitro rumen fermentation efficiency. The ensiling study was randomly assigned according to a $3{\times}3$ factorial arrangement in which the first factor was molasses (M) supplement at 0%, 1%, and 2% of crop dry matter (DM) and the second was urea (U) supplement as 0%, 0.5%, and 1% of the crop DM, respectively. After 28 days of ensiling, the silage samples were collected and analyzed for chemical composition. All the nine Leucaena silages were kept for study of rumen fermentation efficiency using in vitro gas production techniques. The present result shows that supplementation of U or M did not affect DM, organic matter, neutral detergent fiber, and acid detergent fiber content in the silage. However, increasing level of U supplementation increased crude protein content while M level did not show any effect. Moreover, the combination of U and M supplement decreased the content of mimosine concentration especially with M2U1 (molasses 2% and urea 1%) silage. The result of the in vitro study shows that gas production kinetics, cumulation gas at 96 h and in vitro true digestibility increased with the increasing level of U and M supplementation especially in the combination treatments. Supplementation of M and U resulted in increasing propionic acid and total volatile fatty acid whereas, acetic acid, butyric acid concentrations and methane production were not changed. In addition, increasing U level supplementation increased $NH_3$-N concentration. Result from real-time polymerase chain reaction revealed a significant effect on total bacteria, whereas F. succinogenes and R. flavefaciens population while R. albus was not affected by the M and U supplementation. Based on this study, it could be concluded that M and urea U supplementation could improve the nutritive value of Leucaena silage and enhance in vitro rumen fermentation efficiency. This study also suggested that the combination use of M and U supplementation level was at 2% and 1%, respectively.

• 한국가축번식학회지
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• 제20권4호
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• pp.413-421
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• 1997

체외에서 포유동물의 난자와 정자의 배양에 관한 연구는 난자의 성숙과정과 수정현상에 대한 많은 새로운 정보를 제공하였다. 동시에 체외수정의 연구로부터 얻은 결과는 또다른 의문을 제기하였다. 특히 동결융해정액을 이용한 돼지체외수정의 경우 낮은 정자의 침입율과 전핵형성율 및 높은 다정자침입(polyspermy)율은 아직도 해결해야할 문제점으로 남아있다. 돼지난자의 성숙에 관한 연구의 성과는 수정후 낮은 전핵형성율을 개선시켰으나 타동물종에 비하면 아직도 매우 낮게 나타나고 있다. 한편 동결정액의 처리를 위하여 caffeine이나 Ca2+와 같은 물질을 수정용 배지내에 첨가하는 등 수정능력획득의 유기를 위하여 여러 가지 방법이 연구되고 있지만 정자의 침입율은 아직도 낮고, polyspermy의 발생율은 높게 나타내고 있다. 따라서 정자의 침입율을 향상시키고 polyspermy를 억제하기 위하여 난관세포와의 공동배양, 난포액을 첨가한 배양액 내에서 정자의 전배양 및 정자농도의 조절은 매우 효과적인 방법으로 이용되어왔다. 그러나 수정란의 체외생산성 향상과 이와 관련된 연구를 보다 효과적으로 수행하기 위해서는 위에서 지적한 문제에 영향을 미치는 요인에 대한 보다 근본적인 이해가 요구된다.

• Biomaterials and Biomechanics in Bioengineering
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• 제3권3호
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• pp.141-155
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• 2016

Two-dimensional (2D) cell culture and in vivo cancer model systems have been used to understand cancer biology and develop drug delivery systems for cancer therapy. Although cell culture and in vivo model studies have provided critical contribution about disease mechanism, these models present important problems. 2D tissue culture models lack of three dimensional (3D) structure, while animal models are expensive, time consuming, and inadequate to reflect human tumor biology. Up to the present, scaffolds and 3D matrices have been used for many different clinical applications in regenerative medicine such as heart valves, corneal implants and artificial cartilage. While tissue engineering has focused on clinical applications in regenerative medicine, scaffolds can be used in in vitro tumor models to better understand tumor relapse and metastasis. Because 3D in vitro models can partially mimic the tumor microenvironment as follows. This review focuses on different scaffold production techniques and polymer types for tumor model applications in cancer tissue engineering and reports recent studies about in vitro 3D polymeric tumor models including breast, ewing sarcoma, pancreas, oral, prostate and brain cancers.

• 한국식품영양과학회지
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• 제14권1호
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• pp.13-22
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• 1985

최대의 in vitro digestibility를 나타내는 조건에서 가열처리한 오징어, 굴, 새우, 명태 및 김을 대상으로 rat-PER, NPR 및 in vitro apparent digestibility를 측정한 결과와 최근 개발된 computed PER(C-PER) 및 discriminant computed PER(DC-PER) technique을 이용하여 계산된 결과를 비교하여, 수산단백질의 품질을 신속하게 측정할 수 있는 가능성을 검토하였다. 오징어, 새우, 멍태의 C-PER은 rat-PER 보다 낮게 계산되었으나, 김은 높게 계산되었으며, 굴은 거의 비슷하게 계산되었다. 또한 이들 시료의 DC-PER은 rat-PER 및 NPR에 C-PER보다 근접된 결과를 보였으며, 단백효율비(PER)와 소화율이 높은 것으로 알려진 굴은 가공저장 중 발생된 산패의 결과로 rat-PER 및 C-PER이 표준단백질인 ANRC casein 보다 훨씬 낮게 계산되었으나, DC-PER은 높게 계산되었다. In vitro 및 in vitro digestibility가 높은 시료의 단백질 품질평가는 DC-PER이 정확하였으며, 품질변화가 심하게 일어난 시료나, 소화율이 낮은 시료는 C-PER technique이 유리할 것으로 생각되었다.

• Asian-Australasian Journal of Animal Sciences
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• 제12권7호
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• pp.1031-1034
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• 1999

Circulating lymphocytes collected from control and heat-stressed buffaloes were subjected to in vitro culture with glucocorticoids, epinephrine or serotonin and their effect, if any, on the release of intracellular prolactin (PRL) was studied using ELISA and C-ELISA techniques. It was noted from the study that PRL level was higher in lymphocytes than in plasma of the control and heat-stressed animals, and that the PRL levels increased in the plasma of heat-stressed animals compared to that of non stressed animals with a significant decrease in lymphocytic PRL content by heat stress. Epinephrine and serotonin significantly increased the release of intracellular PRL from the lymphocytes of both in the control and the heat-stressed buffaloes but release of PRL from lymphocyte was not significantly changed by cortisol treatment in both control and heat-stressed buffaloes as compared to epinephrine and serotonin in vitro. When lympocytes were incubated with serotonin, it caused drastic lysis of the lymphocytes but epinephirine and cortisol did not show any lysis. It may be concluded from this study that hormones like epinephrine or serotonin known to increase during stress, release intracellular PRL from lymphocytes, the satellite PRL storage/synthesizing organ of blood, although the mechanism of the release is different.