• Clinical and Experimental Reproductive Medicine
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• 제24권1호
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• pp.21-26
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• 1997

본 연구는 체외생산된 생쥐 배반포기배의 ICM 세포에서 EGF-R 발현유무를 immunosurgery와 indirect immunofluorescence (간접 면역 형광방법) 을 이용하여 조사하고자 실시하였다. 본 실험에 사용된 ICM 세포는 체외수정 후 96시간째에 회수된 생쥐 배반포기배를 immunosurgery 방법을 이용하여 얻어졌으며, 회수된 ICM세포는 생사유무와 EGF-R 발현유무 조사에 공시 되어졌다. 그 결과를 요약하면 다음과 같다. ICM세포의 회수율은 rabbit anti-mouse serum (antiserum) 과 guinea pig serum (complement) 에 각각 15-30 분과 15-60분 동안 처리 했을 경우 8.0-84.2% 였으며, 또한 처리시간이 각각 30분과 60분일 때 가장 높은 회수율 (84.2%) 을 얻었다. Immunosurgery 후 얻어진 ICM세포의 생존 유무를 조사하기 위해 live/dead염색 방법을 이용하였던 바, 처리된 ICM세포중 93.8-100%의 생존율을 나타내어 회수된 ICM세포는 유해한 영향을 받지 않았다는 것을 알 수 있었다. 또한, 간접면역 형광방법을 이용하여 ICM세포에서 EGF-R가 발현되는 것을 확인 하였다. 따라서, ICM세포에서의 EGF-R의 발현은 인위적으로 첨가된 EGF의 이용가능성을 높임으로서 체외에서의 착상전 배 발달을 증진시킬 수 있을 것으로 사료된다.

• The Korean Journal of Physiology and Pharmacology
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• 제10권3호
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• pp.123-129
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• 2006

In several experimental models, estrogens protect neurons against ischemic insults. However, the recent clinical studies of hormone replacement showed negative results to prevent stroke. Therefore, optimal models to study estrogen replacement for neuroprotection are needed before its clinical ap-plication. Organotypic hippocampal slice under oxygen-glucose deprivation (OGD) has been established as a model of cerebral ischemia and has advantages to study drug effects. We investigated whether estrogen protected CAI neurons and affected activation of Akt (pAkt) in CAI region under OGD. Thus, rat hippocampal slices on day 7 of culture were treated with $17-{\beta}$ estradiol (E, 1 nM) for 7 days before 30 min OGD, and cell death of CAI neurons was quantified by propidium iodide (PI) staining and expression of pAkt was studied by Western blot and immunofluorescence. PI intensity in slices treated with E was significantly reduced 72 hour after OGD compared to that of non-treated slices (p < 0.05). E pretreatment also increased the expression of pAkt 72 hour after OGD compared to that of no treatment (p<0.01). These data suggest that estrogen pretreatment may rescue neurons from ischemic insults through the activation of Akt and also indicate that our model would be a useful alternative method to study the mechanisms and effects of estrogen replacement treatment for neuroprotection.

• 미생물학회지
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• 제23권1호
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• pp.64-68
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• 1985

페디스토마 성충의 단백질을 정제하여 이를 항원으로 면역시킨 mouse의 spleen cell과 myeloma cell을 세포융합 시킨 결과 융합율은 56% 이었고. 3차 cloning한 후의 항체생산율은 89% 이였으며, 이들 융합세포로 부터 IgM을 생산하는 hybridoma clone과 IgG을 생산하는 hybridoma clone을 얻었다. IgM을 생산하는 hybridoma clone의 supernate는 antibody titer가 희석농도 1 : 64까지 양성이었고. IgG을 생산하는 hybridoma clone의 supernate는 1 : 256까지 양성이었다.

• Clinical and Experimental Reproductive Medicine
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• 제47권3호
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• pp.186-193
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• 2020

Objective: The effectiveness of Staphylococcus protein A (SPA) in improving the penetration ability of sperm and reducing antisperm antibody (ASA) titers in immunologically infertile males was evaluated. Methods: Seminal fluid samples were obtained from 15 infertile men, and ASA titers were assessed with the latex agglutination test. Identification of immunoglobulin (Ig) classes and characterization of the antigens involved in the immune response were performed using indirect immunofluorescence. Local ASAs typically present as a mixture of IgG and IgA classes. The capillary tube penetration method was used to assess the capability of spermatozoa to penetrate the cervical mucus (CM). Results: ASAs associated with the neck region of sperm showed a significantly lower migration distance in the CM of infertile females than ASAs associated with the head or tail segments. ASA-positive seminal fluid exhibited significant increases in the mean migration distance (2.6 ± 1.4 cm vs. 1.54 ± 1.1 cm, respectively; p< 0.001) and sperm concentration (174 ± 121.0 × 10³/mL vs. 101 ± 93.7 × 10³/mL, respectively; p= 0.033) after treatment with SPA compared to pre-treated samples. A significant reduction (p< 0.01) in the recorded ASA titer was detected. Conclusion: These results indicate that SPA can be used as a sorting regimen for insemination programs. However, further studies are warranted to assess its influence on pregnancy rate.

• 대한수의학회지
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• 제34권4호
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• pp.805-813
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• 1994

Immunohistochemical study on the intestinal tissues obtained from the 21 pigs of the 14 terms in Korea in which the clinical and epidemiological features had indicated the possible outbreaks of porcine epidemic diarrhea(PED) was performed using the indirect immunofluorescence test and/or the immunoperoxidase method in order to detect PED viral antigens in the infected cells of the intestines, and histopathological features were described as well. By immunohistochemical analysis, PED viral antigens were detected in the epithelial cells covering the small intestinal villi and recognized slightly in the cells lining the colonic surface epithelium as well. Occasional fluorescence was also seen in a few intestinal crypt epithelium. On light microscopy, the piglets with PED showed marked villous atrophy and fusion, and severe enterocyte degeneration and desquamation. On the other hand, the older pigs more than 4 week old age was mild villous atrophy and fusion, severe villous epithelial cell proliferation, and moderate mononuclear cell infiltration.

• Journal of Korean Biological Nursing Science
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• 제8권1호
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• pp.39-48
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• 2006

Purpose: To see the effects of capsaicin on the peripheral nerve damage of intervibrissal fur in mature rats, Method: 24 female mature rats($200{\sim}250g$) are divided to 3 groups and compared with each other. Immunofluorescence dye using CGRP and PGP antibodies was performed and 8 weeks after administration of capsaicin with control group. Result: The immunopositive reaction against PGP and CGRP was reduced by the damage of epidermal and dermal endings in unmyelinated sheath and thin myelinated sheath and the group after 8weeks showed distinct positive reaction of PGP and CGRP than the group after 4 weeks which means the recover of nerves. Conclusion: As a result, capsaicin influenced on pain-related neurotransmitter like CGRP when administerd to mature rats and even though it caused the damages on unmyelinated sheath and thin myelinated sheath, the damaged nerves recovered after 8 weeks. Also the research about sensory nerve endings scattered over middle dermal and deep epidermal layers such as lanceolate, merkel reticular, Ruffini endings should be studied when the research of the inner conical body is performed. Further studies are necessary about the toxicity and effect of capsaicin on the peripheral nerve endings.

• 한국동물위생학회지
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• 제29권1호
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• pp.1-8
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• 2006

Porcine circovirus (PCV) is a small, nonenveloped virus that contains a single-stranded circular DNA genome of about 1.76 kb and belongs to the family circoviridae. The PCV-2 has been incriminated as the cause of post-weaning multisystemic wasting syndrome (PMWS) , an emerging disease in pigs. In the present study, a PCR assay was applied to detect PCV-2 in tissue samples. The presence of PCV-2 antigen in the porcine tissues was confirmed by indirect immunofluorescence (IIF) with PCV-2 specific monoclonal antibodies. And then DNA extracted from PCV-2 positive tissues was used as a template. One oligonucleotide primer suitable for PCR was selected from a published PCV-2 sequence (Genbank). Amplified PCR product was detected the same fragment lengths of 416 bp as a control. Based on these results, it was suggested that the PCR is a simple and sensitive method for support diagnostic purposes.

• Journal of Microbiology and Biotechnology
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• 제25권4호
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• pp.547-553
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• 2015

The yeast surface-displayed cDNA library has been used to identify unknown antigens. However, when unknown target antigens show moderate-to-low abundance, some modifications are needed in the screening process. In this study, a directional random-primed cDNA library was used to increase the number of candidates for the unknown antigen. To avoid the loss of target yeast clones that express proteins at a low frequency in the cDNA library, a comprehensive monitoring system based on magnetic-activated cell sorting, fluorescence-activated cell sorting, and immunofluorescence was established, and a small number of target yeast cells was successfully enriched. These results showed that our optimized method has potential application for identifying rare unknown antigens of the human monoclonal antibody.

• The Korean Journal of Physiology and Pharmacology
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• 제25권1호
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• pp.79-85
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• 2021

α-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid (AMPA) receptors are differentially regulated in the nucleus accumbens (NAcc) of the brain after cocaine exposure. However, these results are supported only by biochemical and electrophysiological methods, but have not been validated with immunohistochemistry. To overcome the restriction of antigen loss on the postsynaptic target molecules that occurs during perfusion-fixation, we adopted an immersion-fixation method that enabled us to immunohistochemically quantify the expression levels of the AMPA receptor GluA1 subunit in the NAcc. Interestingly, compared to saline exposure, cocaine significantly increased the immunofluorescence intensity of GluA1 in two sub-regions, the core and the shell, of the NAcc on withdrawal day 21 following cocaine exposure, which led to locomotor sensitization. Increases in GluA1 intensity were observed in both the extra-post synaptic density (PSD) and PSD areas in the two sub-regions of the NAcc. These results clearly indicate that AMPA receptor plasticity, as exemplified by GluA1, in the NAcc can be visually detected by immunohistochemistry and confocal imaging. These results expand our understanding of the molecular changes occurring in neuronal synapses by adding a new form of analysis to conventional biochemical and electrophysiological methods.

• 한국동물생명공학회지
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• 제37권2호
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• pp.80-86
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• 2022

The ability to determine the sex of bovine embryos before the transfer is advantageous in livestock management, especially in dairy production, where female calves are preferred in milk industry. The milk production of female and male cattle benefits both the dairy and beef industries. Pre-implantation sexing of embryos also helps with embryo transfer success. There are two approaches for sexing bovine embryos in farm animals: invasive and non-invasive. A non-invasive method of embryo sexing retains the embryo's autonomy and, as a result, is less likely to impair the embryo's ability to move and implant successfully. There are lists of non-invasive embryo sexing such as; Detection of H-Y antigens, X-linked enzymes, and sexing based on embryo cleavage and development. Since it protects the embryo's autonomy, the non-invasive procedure is considered to be the safest. Invasive methods affect an embryo's integrity and are likely to damage the embryo's chances of successful transformation. There are different types of invasive methods such as polymerase chain reaction, detection of male chromatin Y chromosome-specific DNA probes, Loop-mediated isothermal amplification (LAMP), cytological karyotyping, and immunofluorescence (FISH). The PCR approach is highly sensitive, precise, and effective as compared to invasive methods of farm animal embryonic sexing. Invasive procedures, such as cytological karyotyping, have high accuracy but are impractical in the field due to embryonic effectiveness concerns. This technology can be applicable especially in the dairy and beef industry by producing female and male animals respectively. Enhancing selection accuracy and decreasing the multiple ovulation embryo transfer costs.