• Applied Microscopy
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• v.32 no.2
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• pp.91-105
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• 2002

Urea transport in the kidney is mediated by a family of transporter proteins that includes renal urea transporters (UT-A) and erythrocyte urea transporters (UT-B). The cDNA of five isoforms of rat UT-A, UTA1, UT-A2, UT-A3, UT-A4, and UT-A5 have been cloned. The purpose of this study was to examine the expression of UT-A (L194), which marked UT-A1, UT-A2 and UT-A4. Male Sprague-Dawley rats, weighing approximately 200 g, were divided into three group: control rats had free access to water, dehydrated rats were deprived of water for 3 d, and water loaded rats had free access to 3% sucrose water for 3 d before being killed. The kidneys were preserved by in vivo perfusion through the abdominal aorta with the 2% paraformaldehyde-lysine- periodate (PLP) or 8% paraformaldehyde solution for 10 min. The sections were processed for immunohistochemical studies using pre-embedding immunoperoxidase method and immunogold method. In the normal rat kidney, UT-A1 was expressed intensely in the cytoplasm of the inner medullary collecting duct (IMCD) cell and UT-A2 was expressed on the plasma membrane of the terminal portion of the shortloop descending thin limb (DTL) cells (type I epithelium) and of the long-loop DTL cells (type II epithelium) in the initial part of the inner medulla. Immunoreactivity for UT-A1 in the IMCD cells, was decreased in dehydrated animals whereas strongly increased in water loaded animals compared with control animals. In the short-loop DTL, immunoreactivity for UT-A2 was increased in intensity in both dehydrated and water loaded groups. However, in the long-loop DTL of the outer part of the inner medulla, immunoreactivity for UT-A2 was markedly increase in intensity in dehydrated group, but not in water loaded group. In conclusion, in the rat kidney, UT-A1 is located in the cytoplasm of IMCD cells, whereas UT-A2 is located in the plasma membrane of both the short-and long-loop DTL cells. Immunohistochemistry studies revealed that UT-A1 and UT-A2 may have a different role in urea transport and are regulated by different mechanisms.

• Food Science of Animal Resources
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• v.24 no.3
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• pp.283-292
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• 2004

This study was conducted to investigate feeding effects of the high pressure boiled extracts (HPBE) of the Ogol chicken with herbs on glucose, hormones and immunological response (cytokine, specific antibody) of serum in the rat which fed either with normal feed (T$_1$), normal feed + herb HPBE (T$_2$), normal feed + Ogol chicken HPBE (T$_3$), normal feed + mixture of cross-bred Ogol chicken HPBE (T$_4$) hydrolyzed with Flavourzyme 0.1% for 35 days. During experimental period, there was a weak trend to have a higher glucose content for the T$_4$ group with 102.27${\pm}$5.95 mg/dL, but it was not significantly higher than other treatments. For insulin level, T$_1$ group showed numerically a slightly higher level with 6.79${\pm}$4.64 ${\mu}$IU/mL, but the difference was not significant in statistic term due likely to a large variation in comparison with other treatments. The treatments did not significantly alter testosterone level in rat plasma with 1.09, 1.46, 0.98, 1.13 ng/mL in T$_1$, T$_2$, T$_3$ and T$_4$, respectively. T$_4$ treatment increased the aldosterone level to a significantly (p<0.05) higher level (273.33 ng/dL) than other treatments. The extract treated rat showed a tendency in the cortisol level of lower levels than the control group, particularly, it was significantly (p<0.05) lower in T$_3$ group than other groups. T$_3$ and T$_4$ groups showed higher levels for interlukin-4 (IL-4) and anti-BSA IgG in immune cells and plasma. T$_2$, T$_3$ and T$_4$ treatments showed a slightly higher levels in v-interferon (INF-r) than the control, with a greater effect for T4 treatments. These results suggested that HPBE of the cross-bred Ogol chicken hydrolyzed with Flavourzyme increased immunological activity and decreased the concentration of cortisol and aldosterone hormones.

• Proceedings of the Korean Vacuum Society Conference
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• 2012.08a
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• pp.311-312
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• 2012

본 발표에서는 광학적 분석 시스템에 적용 가능한 발광소자(광원)과 수광소자(광센서)를 집적화시키는 모듈(수 발광 집적모듈) 기술을 제시하고자 한다. 이러한 수-발광 집적모듈은 다양한 응용 분야에 적용 될 수 있다. 예를 들어, 광신호 감지를 위한 광통신용 송-수신 모듈(optical communication), 의료/진단 분야에서 단백질/DNA/박테리아 등의 검출 및 분석에 관한 바이오 센서(bio-sensor), 그리고 대기(가스)/수질 모니터링에 관한 환경센서 등 매우 광범위한 분야에 해당되는 요소 기술이라 할 수 있다. 특히, 이들 분야들 중 바이오 물질을 분석하고 검출하는 광학적 바이오 센서 기술은 높은 경제적 가치와 산업적 성장 잠재력으로 인해 오랫동안 활발한 연구가 진행되어 오고 있다. 이러한 광학적 바이오 센서에서 가장 범용적인 방법 중 하나가 항온-항체 면역반응을 기반으로 하는 형광 검출(fluorescence detection) 기법이다. 이러한 시스템은 전체적으로 광원, 광학계, 그리고 센서로 구성되는데 기존에 일반적으로 사용되고 있는 형광 현미경의 경우는 민감도가 우수하다는 장점은 있으나 상당히 고가이고 부피가 크며 복잡한 광학구성으로 이루어져 있다는 한계점을 가지고 있다. 이러한 맥락에서 고민감도를 확보하면서 휴대성, 고속처리, 저가 등의 특성을 가진 시스템에 대한 요구가 갈수록 증가하고 있다. 이를 해결하기 위한 핵심기술 중의 하나가 수-발광 부분을 집적화 시키는 기술이라 할 수 있다. 본 연구에서는 바이오 센서 기술의 하나로서 형광을 측정하여 혈액내의 진단 지표인자를 검출할 수 있는 휴대용 혈액진단기기에 적용되는 소형 수 발광 집적 모듈을 개발하였다. 혈액내의 검출 성분의 양에 따라 형광의 세기가 변화하게 됨으로써 정량적인 검출이 가능한 원리이다. 모듈의 구조는 크게 광원(발광소자), 광학계, 그리고 광센서(수광소자) 세 영역으로 나누어 진다. 광원은 635 nm 적색 레이저다이오드로서 형광체(Alexa Fluor 647/발광파장: 668 nm)를 여기 시키는 기능을 하며 장착된 볼렌즈 의해 샘플의 형광체 영역으로 집광된다. 광학계는 크게 시준렌즈(collimating lens)와 광학필터로 구성됨으로써 샘플로부터 발생되는 광을 적절하게 수광소자로 전달하는 기능을 하게 된다. 여기서 광학필터의 경우는 기본적으로 Distributed Bragg's Reflector(DBR) 구조로써 실리콘(Si) 포토다이오드 상부에 모노리식(monolithic)하게 형성되며 검출 샘플로부터 진행되는 레이저 광(잡음의 주원인)은 차단하고 형광(광신호)만 통과 시키는 기능을 하게 된다. 따라서 신호 대 잡음비(S/N ratio)를 향상시키기 위해서는 정밀한 광 필터링 기능이 요구됨으로써 박막의 세밀한 공정 조건과 구조적-광학적 특성 분석이 수행되었다. 마지막으로 포토다이오드 소자는 일반적인 구조 이외에 중앙에 원형 구멍이 형성된 특별한 구조가 적용된다. 이것은 포토다이오드 구조에 변화를 줌으로써 모듈 구조를 효율적으로 응용할 수 있다는 의미를 갖는다. 또한 포토다이오드의 전기적-광학적 측정 분석을 통해 잡음 및 감도 특성이 세부적으로 조사되며 형광신호를 효과적으로 측정할 수 있음을 확인하였다. 최종적으로 제작된 모듈은 약 $1{\times}1{\times}1cm^3$ 내외 정도의 크기를 갖는다. 요약하자면 본 발표에서는 광학적 바이오센서에 적용할 수 있는 소형 수-발광 소자 집적모듈을 소개한다. 전체 모듈 설계는 최소한의 부피를 가짐과 동시에 측정의 정밀성을 향상시키는데 초점을 맞추어 진행하였다. 세부요소인 광학필터와 포트다이오드의 경우 잡음 및 민감도에 미치는 중요성 때문에 세밀한 공정 및 특성분석이 수행되었다. 결론적으로 독자적인 설계 및 공정을 통해 휴대성 및 정밀성 등의 목적에 부합한 경쟁력 있는 수-발광 소자 집적모듈 제작 기술을 확보하였다.

• Journal of Life Science
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• v.14 no.3
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• pp.375-384
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• 2004

Fibroblast growth factors (FGFs) are known to induce multiple functions in early development, including mesoderm formation, gastrulation movement and antero-posterior patterning. The induction of mesoderm from Xenopus presumptive ectoderm and the combination effect on inducing organs of bFGF(basic FGF) and HGF (Hepatocyte Growth Factor) were studied. Explants were cultured in the combined solution for 3 days to normal embryo arrive at St. 43. These effects on combined dose were examined by histological experiment and by immunohistochemical method. The concentrations of growth factors were tested in 0, 0.5, 1, 10 and also tested in 50 ng/ml of bFGF, and 0, 1, 10, 50 and 100ng/ml of HGF respectively. The synergistic effects were seen in the combined-dose of bFGF and HGF rather than in single dose. Various organs were differentiated and highest inducing effects were seen at the combined concentration of 1 ng/ml of bFGF and 10ng/ml of HGF, and at the concentration 10ng/ml of bFGF and 1 ng/ml of HGF. The bFGF induces various organs from cultured animal cap explants and the effects are time and dose-dependent. HGF is also a potent mitogen for renal tubular cells and for mature hepatocytes in primary culture. Eyes were developed in high percentage at the combined concentration of 1 and 10ng/ml of bFGF, and 1 and 10 ng/ml of HGF. From the induced eye and normal embryonic eye, RPE65 was commonly detected by monoclonal antibodies 40All and 25F5 and the localization of RPE65 was seen by AP reaction.

• Pediatric Infection and Vaccine
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• v.23 no.2
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• pp.149-154
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• 2016

Although an association of Kawasaki disease (KD) with infectious agents has been suggested, none have been proven to cause KD. In this case study, we present a case of KD with concurrent onset of influenza and Mycoplasma pneumoniae (MP) infections. A 27-month-old boy presented with prolonged fever, cough, and rhinorrhea. During the initial testing, influenza A infection was identified, and he was treated with oseltamivir. Despite the antiviral therapy, the fever persisted, and he had cervical lymph node enlargement, bilateral conjunctival injection, fissured red lips, strawberry tongue, and erythematous skin lesions on the Bacillus Calmette-$Gu{\acute{e}}rin$ vaccination site. Thus, the patient was diagnosed with KD and was treated with intravenous immunoglobulin (IVIG). The result of the initial antimycoplasma immunoglobulin M (IgM) antibody testing and was positive, and an increased IgM titer from baseline was found in a repeat test. We reviewed the hypotheses on pathogens known to be associated with KD and the etiology of KD. Based on our findings, we suspect that symptoms of KD and coronary artery lesions can occur from various infections besides those caused by Mycoplasma species and influenza viruses.

• Childhood Kidney Diseases
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• v.6 no.1
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• pp.56-60
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• 2002

Purpose Streptococcus pneumoniae is a major pathogen in both adults and children, causing significant morbidity and mortality In patients with nephrotic syndrome, Streptococcus pneumoniae is a major cause of spontaneous peritonitis, and the increasing incidence of penicillin-resistance strain facilitates the development of effective vaccine. The limitation of current pneumococcal polysaccharide vaccine prompted development of polysaccharide- protein conjugate vaccine. Methods: We reviewed the medical record of total 225 steroid responsive nephrotic patients to ascertain the effectiveness of 23- valent pneumococcal polysaccharide vaccine. Results. Twenty- eight patients have developed peritonitis during the courses, and 7 of those have recurrent peritonitis. Fifty- five patients were vaccinated and followed- up for 1- 108 months (mean 38.5 months), and during the follow- up period, pneumococcus related peritonitis was not detected. Vaccine- related relapse of nephrotic syndrome w as absent. Conclusion: In spite of the non- consensus about the efficacy of PPV23, clinically it benefits, and until the clinical trial of PCV7 is completed, PPV23 will be recommended. (J Korean Soc Pediatr Nephrol 2002;6: 56-60)

• Korean Journal of Head & Neck Oncology
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• v.13 no.2
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• pp.169-179
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• 1997

서론 : Laminin의 수용기로 알려진 Integrin $\alpha6\beta4$의 세포내 표현 정도는 편평상피암을 위시한 여러 악성종양의 전이능력 및 예후와 밀접한 상관관계가 없다고 알려져 있다. 이 Integrin은 Laminin과 같은 세포와 리간드와 결합하면 상피세포의 기저막 지주 구조물인 hemidesmosome의 세포체질 요소(cytoskeletal element)와 연관되어 그 결과 세포의 기저막과 세포내 케라틴을 연결하는 역할을 한다. Integrin $\alpha6\beta4$는 구조적으로 다른 많은 integrin들과 달리 $\beta$4의 세포질내 영역(cytoplasmic domain)이 특징적으로 크다. 이 세포질내 영역 $\beta$4 integrin의 기능은 아직 밝혀지지 않고 있으나 아마 세포 성장의 신호전달 및 악성종양의 특징인 침윤 전이에 관련할 것으로 보아지고 있다. 재료 및 방법: 저자들은 우선 $\beta$4 integrin의 wild type s-DNA와 $\beta$4 세포질내 영역(cytoplasmic domain) 및 $\beta$4의 tyrosine 인산화 반응 부위가 각각 결손된 c-DNA를 PCR을 통하여 합성하여 pRc/CMV 벡터에 삽입한 후 원래 $\beta$4 integrin의 발현이 결집된 인간 방광암 세포에 Calcium phosphate precipitation 방법으로 주입(transfection)시켜 형질변환된 세포를 면역형광법, Flow cytometry 및 Immunoprecipitation 방법으로 클로닝하여 wild type $\beta$4-full length(Clone FL), truncated $\beta$4-cytoplasmic domain(C1one CD), 및 mutated $\beta$4-tyrosine phosphorylation site (Clone M)을 얻었다. 암 세포의 부착 및 침투 능력의 기능적 연구로 모노 클로날 항체와 fibronectin, laminin, Matrigel을 단백질 기질로 사용하였으며 결과 비교를 위하여 pRc/CMV 벡터만 주입시켰던 클로운과 방광암 세포주를 $\beta$4 integrin 음성 대조군으로 또한 이 Integrin의 높은 발현을 보이는 두경부 편평상피암 세포주를 양성 대조군으로 이용하였다. 결과 : 세포부착능력에 있어서 온전한 $\beta$4 cytoplasmic domain이 존재하는 클로운이 laminin에 강한 부착능력을 보였으나 fibronectin의 부착정도는 $\beta$4 integrin의 표현정도와 관계없이 모든 클로운에서 비슷하였다. Matrigel을 투과하는 암세포 침윤 능력에서는 $\beta$4 integrin의 표현이 존재하는 클로운들이 투과 능력이 높았으나 세포외 리간드가 없는 control membrane을 사용하였을 때와 비교하여 투과능력의 차이를 보이지 않았다. 결론 : 유전자 주입(transfection) 방법으로 integrin의 다양한 클로운의 합성이 가능하여 이 Integrin의 암 세포의 부착 및 침투 능력에서의 기능을 규명 할 수 있게 한다. $\beta$4 integrin은 편평상피 암세포의 부착에 있어서 세포외 리간드 laminin과 특이 결합하여 부착 능력을 높이는 중요한 역할을 하며 편평상피 암세포의 침투에 있어서는 $\beta$4 integrin의 표현이 침투 능력을 높이는 역할을 하나 이때에는 laminin과 같은 리간드와의 특이 결합에 의존하지는 않는 것으로 사료된다.

• Food Science and Preservation
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• v.23 no.5
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• pp.666-672
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• 2016

The objective of this study was to evaluate the effects of low temperature-adapted Saccharomyces cerevisiae Y297 and fermentation temperatures on the quality of Yakju brewed. Physicochemical properties of Yakju brewed were compared pH, total acidity, ethanol, free amino acid, organic acid contents, and volatile flavor compounds in S. cerevisiae Y297 with control treatment. Cooked non-glutinous rice and saccharogenic amylase in koji were mixed with ethanol-producing yeasts and then fermented at $15^{\circ}C$, $20^{\circ}C$, and $25^{\circ}C$ for 20 days. Yakju brewed using the Y297 treatment showed the highest ethanol yield (17.9%) at $20^{\circ}C$. Expression of heat shock protein (HSP) 104 was evaluated by immunoblotting as an indication of adaptation to low temperatures ($15^{\circ}C$); levels of the HSP104 protein were higher in the Y297 treatment than in the control. Organic acid analysis showed that the lactic acid content of Yakju brewed using the control was the highest at $25^{\circ}C$. Finally, free amino acid analysis showed that the Y297 treatment had a higher proportion of essential amino acids than the control. Overall, these results indicate that S. cerevisiae Y297 could be used as a suitable yeast for Yakju brewed under low temperature ($15^{\circ}C$) condition.

• Tuberculosis and Respiratory Diseases
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• v.41 no.5
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• pp.484-488
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• 1994

Background : The antigen-specific receptor on the surface of most peripheral T lymphocytes is a disulfide-linked heterodimer composed of $\alpha$ and $\gamma$ subunits, noncovalently associated with CD3 polypeptides. Recently, a novel type of CD3-associated heterodimer was described on a T cell subset that does not express CD4 or CD8 molecules. This second type of TCR dimer is composed of chains encoded for by the $\gamma$- and $\delta$-TCR genes. These cells may exert both cytotoxic and lymphokine producing functions. Although it was reported that some ${\gamma}{\delta}$-TCR might recognize an MHC-linked determinant, the funεtion or physiologic ligand for this new receptor is not yet clear. It was found that ${\gamma}{\delta}$-TCR can react with 65 kD heat shock protein of M. tuberculosis, which suggests the possible protective role of ${\gamma}{\delta}$ T lymphocytes against tuberculosis. In our previous study, there was neither the increase in number nor the functional activation of ${\gamma}{\delta}$ T cells in the peripheral blood from patients with pulmonary tuberculosis. Now we report the distribution of ${\gamma}{\delta}$ T cells in the regional sites of M. tuberculosis infection, especial1y tuberculous lymphadenitis. Methods : Lymph nodes from patients with pathologically-proven tuberculous lymphadenopathy (n=5) and reactive hyperplasia (n=3) were used. Tissues were frozen in liquid nitrogen immediately after removal and stored below $-70^{\circ}C$. The cryostat sections of these frozen specimens were stained with anti-Leu-4 Ab, Identi-T TCR ${\delta}1$, and Identi-T ${\beta}F1$. The number of positively stained cells were counted at high power field. Results : The infiltration of ${\gamma}{\delta}$ T cells was significantly higher in the lymph nodes from patients with tuberculous lymphadenopathy than that with reactive hyperplasia ($16.3{\pm}10.3%$ vs. $1.7{\pm}1.5%$). Conclusion : These results suggest that ${\gamma}{\delta}$) T cells may play a role in the defense against M. tuberculosis infection, especially in the regional sites of infection.

• Journal of the Korean Society of Food Science and Nutrition
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• v.24 no.3
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• pp.470-486
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• 1995

Mammary epithelial cells contain a subpopulation of cells with a large proliferativ potential which are responsible for the maintenance of glandular cellularity and are the progenitor cells of mammary cancer. These clonogens give rise to multicellular clonal alveolar or ductal units(AU or DU) on transplantation and hormonal stimulation. To isolate putative mammary clonogens, enzymatically monodispersed rat mammary epithelial cells from organoid cultures and from intact glands are sorted by flow cytometry according to their affinity for FITC labeled peanut lectin(PNA) and PE labeled anti-Thy-1.1 antibody(Thy-1.1) into four subpopulations : cells negative to both PNA and Thy-1.1(B-), PNA+cells, Thy-1.1+cells, and cells positive to both reagents(B+). The in vivo transplantation assays indicate that the clonogenic fractions of PNA+cells from out-growths of organoids in primary cultures for three days in complete hormone medium(CHM) are significantly higher than those of cells from other subpopulations derived from cultrues or from intact glands. Extracellular matrix(ECM) is a complex of several proteins that regulated cell function ; its role in cell growth and differentiation and tissue-specific gene expression. It can act as a positive as well as a negative regulator of cellular differentiation depending on the cell type and the genes studied. Regulation by ECM is closely interrelated with the action of other regulators of cellular function, such as growth factors and hormones. Matrigel supports the growth and development of several different multicellular colonies from mammary organoids and from monodispersed epithelial cells in culture. Several types of colonies are observed including stellate colonies, duct-like structures, two- and three-dimensional web structures, squamous organoids, and lobulo-duct colonies. Organoids have the greatest proliferative potential and formation of multi-cellular structures. Phase contrast micrographs demonstrate extensive intracellular lipid accumulation within the web structures and some of duct-like colonies. At the immunocytochemical and electron micrograph level, casein proteins are predominantly localized near the apical surface of the cells or in the lumen of duct-like or lobulo-duct colonies. Squamous colonies are comprised of several layers of squamous epithelium surrounding keratin pearls as is typical fo squamous metaplasia(SM). All-trans retinoic acid(RA) inhibits the growth of SM. The frequency of lobulo-ductal colony formation increased with the augmentation of RA concentration in these culture conditions. The current study models could provide powerful tools not only for understanding cell growth and differentiation of epithelial cells, but also for the isolation and characterization of mammary clonogenic stem cells.