• Food Science and Biotechnology
• /
• v.18 no.2
• /
• pp.317-322
• /
• 2009

The proteins from 15 types of cultivar of hot peppers cultivated in Korea were extracted and its allergenicity was investigated by immunoblotting and enzyme-linked immunosorbent assay (ELISA). The immunoblotting of hot pepper proteins extracts (HPEs) against serum of hot pepper sensitized patients revealed dominant IgE binding to 14, 37, and 40 kDa molecules. The specific levels of IgE to HPEs sample No. 1, 3, and 7 were much higher than the other samples in patients. Also, IgE binding capacity of HPEs were not reduced by thermal processing and digestion in ELISA using human IgE antibody acquired from hot pepper sensitized patients. By means of Western blotting using anti-thaumatin IgY, thaumatin-like protein (TLP) acting as allergen in several plants and fruits was detected in tested hot peppers. This study demonstrates that the antigenic protein in hot peppers are present but are differently contained according to cultivars.

• Genomics & Informatics
• /
• v.17 no.1
• /
• pp.7.1-7.10
• /
• 2019

Cysticercosis, a parasitic disease caused by Taenia solium metacestode (TsM), has a major global public health impact in terms of disability-adjusted life years. The parasite preferentially infects subcutaneous tissue, but may invade the central nervous system, resulting in neurocysticercosis (NC). NC is an important neglected tropical disease and an emerging disease in industrialized countries due to immigration from endemic areas. The prevalence of taeniasis in Korea declined from 0.3%-12.7% during the 1970s to below 0.02% since the 2000s. A survey conducted from 1993 to 2006 revealed that the percentage of tested samples with high levels of specific anti-TsM antibody declined from 8.3% to 2.2%, suggesting the continuing occurrence of NC in Korea. Modern imaging modalities have substantially improved the diagnostic accuracy of NC, and recent advances in the molecular biochemical characterization of the TsM cyst fluid proteome also significantly strengthened NC serodiagnosis. Two glycoproteins of 150 and 120 kDa that induce strong antibody responses against sera from patients with active-stage NC have been elucidated. The 150 kDa protein showed hydrophobic-ligand binding activities and might be critically involved in the acquisition of host-derived lipid molecules. Fasciclin and endophilin B1, both of which play roles in the homeostatic functions of TsM, showed fairly high antibody responses against calcified NC cases. NC is now controllable and manageable. Further studies should focus on controlling late-onset intractable seizures and serological diagnosis of NC patients infected with few worms. This article briefly overviews diagnostic approaches and discusses current issues relating to NC serodiagnosis.

• Journal of Microbiology and Biotechnology
• /
• v.13 no.6
• /
• pp.912-918
• /
• 2003

Mitotic centromere-associated kinesin (MCAK), which is a novel kinesin with a central motor domain, is believed to playa role in mitotic segregation of chromosome during the M phase of the cell cycle. In the present study, it is shown that a rabbit polyclonal antibody has been produced using the N-terminal region (187 aa) of human MCAK expressed in E. coli as the antigen. To express the N-terminal region in E. coli, the MCAK cDNA fragment encoding N-terminal 187 aa was obtained by PCR and was then inserted into the pET 3d expression vector. Molecular mass of the N-terminal region overexpressed in the presence of IPTG was 23.2 kDa on SDS-PAGE, and the protein was insoluble and mainly localized in the inclusion body that could be easily purified from the other cellular proteins. The N-terminal region was purified by electro-elution from the gel after the inclusion body was resolved on the SDS-PAGE. The antiserum obtained after tertiary immunization with the purified protein specifically recognized HsMCAK when subjected to Western blot analysis, and showed a fluctuation of the protein level during the cell cycle of human Jurkat T cells. Synchronization of the cell-cycle progression required for recovery of cells at a specific stage of the cell cycle was performed by either hydroxyurea or nocadazole, and subsequent release from each blocking at 2, 4, and 7 h. Northern and Western analyses revealed that both mRNA and protein of HsMCAK reached a maximum level in the S phase and declined to a basal level in the G1 phase. These results indicate that a polyclonal antibody raised against the N-terminal region (187 aa) of HsMCAK, overexpressed in E. coli, specifically detects HsMCAK (81 kDa), and it can analyze the differential expression of HsMCAK protein during the cell cycle.

• The Korean Journal of Physiology and Pharmacology
• /
• v.6 no.3
• /
• pp.165-169
• /
• 2002

We have previously demonstrated that phytoestrogens isolated from safflower seeds significantly attenuated bone loss in ovariectomized rats, and directly stimulated proliferation and differentiation of cultured osteoblastic cells. In an attempt to elucidate underlying cellular mechanisms, in the present study we investigated effects of $17{\beta}-estradiol\;(E_2)$ and phytoestrogens such as matairesinol and acacetin, a type of lignan and flavonoid, respectively, on activation of mitogen activated protein (MAP) kinases, extracellular signal-regulated kinase 1 (ERK1) and ERK2, in cultured osteoblastic ROS 17/2.8 cells. Western blot analysis with anti-MAP kinase antibody showed that a wide range concentrations $(10^{-14}\;to\;10^{-6}\;M)\;of\;E_2$ as well as both phytoestrogens induced rapid and transient activation of ERK1/2 through phosphorylation within minutes. Maximum activation of MAP kinases by $E_2$ and phytoestrogens were observed at 10 and 15 min, respectively. $E_2-induced$ phosphorylation of ERK1/2 returned to the control level at 30 min, whereas phytoestrogen-induced phosphorylation was maintained at high level until 30 min. PD-98059, a highly selective inhibitor of MAP kinase, prevented phosphorylation of ERK1/2 in the cells treated either with $E_2$ or phytoestrogens. To examine a possible involvement of estrogen receptor in the activation process of MAP kinase, Western blot analysis was performed in the presence and absence of the estrogen receptor antagonists, ICI 182,780 and tamoxifen. These antagonists blocked MAP kinase phosphorylation induced not only by $E_2,$ but also by the phytoestrogens. To the best our knowledge, this study is the first to demonstrate that phytoestrogens such as flavonoid and lignan extracted from safflower seeds produce a rapid activation of MAP kinase, at least partially via membrane estrogen receptor of the cultured osteoblastic cells.

• Rapid Communication in Photoscience
• /
• v.4 no.2
• /
• pp.48-49
• /
• 2015

For myelination, Schwann cells and neuron cells from dorsal root ganglion (DRG) of rat embryos (E16) were cultured in vitro system. The purified DRG cells with anti-mitotic agents and purified Schwann cells were cocultured and then accomplished myelination processing. Treatment of M. leprae-specific phenolic glycolipid-1 (PGL-1) into this coculture system was performed and then accomplished demyelination. Therefore, we identified demyelination processing using antibody of myelin basic protein (MBP).

• Biomedical Science Letters
• /
• v.17 no.3
• /
• pp.197-202
• /
• 2011

Mitosis count is one of the most helpful morphologic features for distinguishing pulmonary typical carcinoid (TC) from atypical carcinoid (AC). However, identifying areas of highest mitotic activity is tedious and time-consuming, and mitosis count may vary substantially among pathologists. Anti-phosphohistone H3 (PHH3) is an antibody that specifically detects histone H3 only when phosphorylated at serine 10 or serine 28, an event that is concurrent with mitotic chromatin condensation and not observed during apoptosis. In this study, immunohistochemical staining for PHH3 was performed to determine whether PHH3 was a reliable and objective mitosis-specific marker for pulmonary carcinoid tumors. Seventeen cases of surgically resected pulmonary carcinoid tumors (12 TCs and 5 ACs) were obtained and classified according to the 2004 World Health Organization classification. Mitotic counts determined by PHH3 correlated to ones determined by hematoxylin and eosin (H&E) staining; however, PHH3 mitotic counts (mean mitotic counts: 1 in TCs and 3.2 in ACs) were slightly higher than H&E mitotic counts (mean mitotic counts: 0.25 in TCs and 1.8 in ACs). The mitotic counts determined by experienced observer were more correlated to those determined by inexperienced observer with the PHH3-based method (R=0.968, P<0.001) rather than H&E staining (R=0.658, P<0.001). These results suggest that the PHH3 mitotic counting method was more sensitive and simple for detecting mitoses compared to traditional H&E staining. Therefore, PHH3 immunohistochemistry may contribute to more accurate and reproducible diagnosis of pulmonary carcinoid tumors and may be a valuable aid for administrating appropriate clinical treatment.

• Clinical and Experimental Pediatrics
• /
• v.57 no.10
• /
• pp.457-460
• /
• 2014

A flaccid tetraparesis in Bickerstaff's brainstem encephalitis (BBE) is presumed to be a sign of overlapping Guillain-Barr$\acute{e}$ syndrome (GBS). In addition, BBE and Fisher syndrome, which are clinically similar and are both associated with the presence of the immunoglobulin G anti-GQ1b antibody, represent a specific autoimmune disease with a wide spectrum of symptoms that include ophthalmoplegia and ataxia. A 2-year-old boy presented with rapidly progressive ophthalmoplegia, ataxia, hyporeflexia, weakness of the lower extremities, and, subsequently, disturbance of consciousness. He experienced bronchitis with watery diarrhea and had laboratory evidence of recent infection with Epstein-Barr virus (EBV). He was diagnosed as having overlapping GBS and BBE associated with EBV and received treatment with a combination of immunoglobulin and methylprednisolone, as well as acyclovir, and had recovered completely after 3 months. In addition, he has not experienced any relapse over the past year. We suggest that combinations of symptoms and signs of central lesions (disturbance of consciousness) and peripheral lesions (ophthalmoplegia, facial weakness, limb weakness, and areflexia) are supportive of a diagnosis of overlapping GBS and BBE and can be helpful in achieving an early diagnosis, as well as for the administration of appropriate treatments.

• Journal of Microbiology and Biotechnology
• /
• v.19 no.2
• /
• pp.155-160
• /
• 2009

Mast cells and basophils perform important functions as pivotal effector cells in IgE-mediated allergic reactions. KU812F cells, a human basophilic cell line isolated originally from chronic myelocytic leukemia, express a high affinity receptor of IgE, $Fc{\varepsilon}RI$. Kaempferol was extracted and isolated from a methanolic extract of flavonoid-rich Nelumbo nucifera stamens. In the present study, the inhibitory effects of kaempferol on $Fc{\varepsilon}RI$ expression in human basophilic KU812F cells was examined. Flow cytometric analysis revealed that $Fc{\varepsilon}RI$ expression on the cell surface was suppressed in a concentration-dependent manner when the cells were cultured with kaempferol. Moreover, RT-PCR analysis showed that the mRNA levels for $Fc{\varepsilon}RI$ ${\alpha}-\;and\;{\gamma}$-chains were reduced as the result of kaempferol treatment in a concentration-dependent manner. Kaempferol showed its suppressive effects on intracellular $Ca^{2+}$ concentration and histamine release from anti-$Fc{\varepsilon}RI$ ${\alpha}$-chain antibody-stimulated cells in a concentration-dependent manner. These observations indicate that kaempferol may exert antiallergic effect via down regulation of $Fc{\varepsilon}RI$ expression and degranulation.

• Food Science of Animal Resources
• /
• v.20 no.4
• /
• pp.303-310
• /
• 2000

In vitro 소화시 계란 성분이 anti-Y ruckeri IgY 항세 활성을 안정화시키는데 어떡해 관여하는지를 SDS-PAGE와 ELISA로 조사한 결과는 다음과 같다. Anti-Y ruckeris IgY 항체와 난황 및 난백으로 혼합 후 펩신으로소화시킨 후 경우 1시간 후 난황 및 난백이 혼합된 시료에서 모두 anti-Y ruckeri IgY 항체의 heavy chain과 light chain이 분해는 되었지만 어느 정도 밴드를 관찰할 수있었으며, anti-Y, ruckeri IgY 항체 활성 측정한 결과 난황시료는 35%, 난백시료는 61%의 항체 활성을 유지시켰다. Anti-Y ruckeri IgY 항체와 오보알부민, 오보뮤우신, 라이소자임 및 오보뮤코이드를 혼합 한 후 펩신으로 소화시킨 경우 1시간 우에는 anti-Y, ruckeri IgY항체의 heavy chain의 밴드르 fdjsm 정도 볼 수 있는 형태로 나타났으며, anti-Y, ruckeri IgY 항체의 light chain는 오보뮤우신 및 오보뮤코이드가 홉합되어 펩신으로 소화시킨 경우 1시간 후에는 밴드를 거의 볼수 없었으나 오보알부민 오보트란스훼린 및 라이소자임에 혼합된 시료에서 밴드를 관찰할수 있었다. 특히 라이소자임과 오보트란스훼린의 경우 펩신 호화 2시간 우에도 anti-Y, ruckeri IgY 항체의 light 밴드가 관찰되었다. 펩신소화 1시간 후에 난백성분이오보알부민, 오보뮤우신, 라이소자임, 오보트란스웨린 및 오부뮤코이드 중에서 단지 오보트란스훼린만이 38%의 anti-Y, ruckeri IgY 항체 활서을 보인후 2시간 후에도 15% 정동의 활성을 나타내었다. anti-Y, ruckeri IgY항체와 난백 및 전란을 혼합한 다음 무지개송어 위 추출액으로 소화시 2시간 후에 난백은 14%, 전란은 69%로 anti-Y, ruckeri IgY 항체 활성을 유지시킨 것으로 나타났다.X> $e_{I}$ WPi_BE_QE]]]]]로 상징하며 WLWQ에 적용되는 몇 가지 제약을 관찰하고 이를 일반적인 언어원리로 설명한다. 첫째, XP는 주어로만 해석되는데 그 이유는 XP가 목적어 혹은 부가어 등 다른 기능을 할 경우 생략 부위가 생략의 복원 가능선 원리 (the deletion-up-to recoverability principle)를 위배하기 때문이다. 둘째, WLWQ가 내용 의문문으로만 해석되는데 그 이유는 양의 공리(the maxim of quantity: Grice 1975) 때문이다. 평서문으로 해석될 경우 WP에 들어갈 부분이 XP의 자질의 부분집합에 불과하므로 명제가 아무런 정보제공을 하지 못한다. 반면 의문문 자체는 정보제공을 추구하지 않으므로 앞에서 언급한 양의 공리로부터 자유롭다. 셋째, WLWQ의 XP는 주제어 표지 ‘는/-은’을 취하나 주어표지 ‘가/-이’는 취하지 못한다(XP-는/-은 vs. XP-가/-이). 이는 IP내부 에 비공범주의 존재 여부에 따라 C의 음운형태(PF)가 시성이 정해진다는 가설로 설명하고자 했다. WLWQ에 대한 우리의 논의가 옳다면, 본 논문은 다음과 같은 이론적 함의를 기닌다. 첫째, WLWQ의 존재는 생략에 대한 두 이론 즉 LF 복사 이론과 PF 삭제 이론 중 전자의 입장을 지지한다. 둘째, WP를 XP로부터 복원할 때 부분 자질만 복사된다. 이는 어휘가 통사층위로 들어온 이후에도 어휘 자질들이 완전히 동결되는 것이 아니라 계속 지시될 수 있다는 가설을 지지한다.

• The Korean Journal of Zoology
• /
• v.27 no.2
• /
• pp.103-116
• /
• 1984

The structural gene of rabbit hemoglobin was cloned into Pst I site of pBR322 in E. coli. The complementary DNA (cDNA) was synthesized from rabbit globin mRNA with avian myeloblastosis viral reverse transcriptase, and then RNA was destroyed at pH 11. The double stranded cDNA was synthesized with both Klenow fragment of E. coli DNA polymerase I and reverse transcriptase and then the hairpin loop was opened with Sl nuclease. Double stranded cDNA was subsequently tailed with dCTP and annealed to dGMP-tailed vector DNA. After transformation and initial screening of appropriate clones by plasmid size, the cloned colonies were identified by in situ colony hybridization using by plasmid size, the cloned colonies were identified by in situ colony hybridization using $[^32P]$-labeled cDNA probes and characterized the inserts with restriction endonucleases. The expression of cloned globin gene was investigated by standard radioimmunoassay using rat anti-rabbit Hb serum as primary antibody and goat antirat IgG serum as secondary antibody. The result suggested that the chimeric proteins (the part of $\\beta$-lactamase from the vector pBR322 and globin from rabbit) were supposedly produced in E. coli and the product had the antigenic determinant of rabbit hemoglobin.