• BMB Reports
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• v.43 no.9
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• pp.604-608
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• 2010

Tumor necrosis factor-$\alpha$ (TNF-$\alpha$) mediates proinflammatory responses in primary human umbilical vein endothelial cells (HUVECs), and it upregulates the expression of secretory group IIA phospholipase $A_2$ ($sPLA_2$-IIA). $sPLA_2$-IIA plays a pivotal role in inflammation, and antithrombin (AT) possesses properties that are beneficial to endothelial cells. Therefore, we investigated the effects of AT on the expression of $sPLA_2$-IIA in TNF-$\alpha$-stimulated HUVECs. TNF-$\alpha$ potently upregulated the expression of $sPLA_2$-IIA, and prior treatment of cells with AT inhibited the expression of $sPLA_2$-IIA in HUVECs. Also, antibodies or siRNA for syndecan-4 blocked the protective effect of AT. Furthermore, PI3-kinase and the AKT pathway are significantly involved in the AT-mediated inhibition of the expression of $sPLA_2$-IIA. These results show that AT effectively suppresses the upregulated $sPLA_2$-IIA expression, which might contribute to the cytoprotective effects of AT in the treatment of severe inflammatory diseases.

• Biomolecules & Therapeutics
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• v.8 no.2
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• pp.107-112
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• 2000

To investigate the alteration of airway mucin in airway disease patients, immunoassay procedures were employed using monoclonal antibodies HM02 and HM03 (Hybridoma, 18,457-463, 1999). Alteration of mucin release was determined by ELISA and the integrity of mucin was determined by Western blot. In ELISA, it was found that mucin release increased from pneumonia, chronic cough, bronchiectasis, eosinophilic pneumonia, lung cancer and bronchial asthma patients. In Western blot, the increase in immunoreactivity was observed in case of pneumonia, chronic cough, bronchiectasis and bronchial asthma. In bronchial asthma, there was no obvious degradation of mucin while in other diseases, varying degree of mucin degradation was observed. The data from the present study implicate that HMO2 and HM03 are suitable for the immunological analysis of mucin in airway disease patients. The role of increased mucin release and varying degree of mucin degradation on airway diseases should be further investigated in the future.

• Journal of Plant Biotechnology
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• v.36 no.4
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• pp.309-319
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• 2009

Transgenic plant cell cultures for the production of biopharmaceuticals including monoclonal antibodies, recombinant proteins have been regarded as an alternative platform in addition to traditional microbial fermentation and mammalian cell cultures. Plant-made pharmaceuticals (PMPs) have several advantages such as safety, cost-effectiveness, scalability and possibility of complex post-translational modifications. Increasing demand for the quantity and diversity of pharmaceutical proteins may accelerate the industrialization of PMP technology. Up to date, there is no plant-made recombinant protein approved by USFDA (Food and Drug Administration) for human therapeutic uses due to the technological bottlenecks of low expression level and slight differences in glycosylation. Regarding expression levels, it is possible to improve the productivity by using stronger promoter and optimizing culture processes. In terms of glycosylation, humanization has been attempted in many ways to reduce immune responses and to enhance the efficacy as well as stability. In this review article, all these respects of transgenic plant cell cultures were summarized. In addition, we also discuss the general characteristics of plant cell suspension cultures related with bioreactor design and operation to achieve high productivity in large scale which could be a key to successful commercialization of PMPs.

• The Journal of the Korean Society for Microbiology
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• v.35 no.2
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• pp.141-147
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• 2000

We purified enolase from Candida albicans KNIH10 strain which was isolated from a clinical specimen in Korea. The purified enolase was used to detect anti-Candida antibodies in sera of patients with invasive candidiasis. For purification of enolase from the crude extract prepared by French pressure at 20,000 PSI, the fast performance liquid chromatography (FPLC) using DEAE-sepharose column was used. The elutes at $0.3{\sim}0.4\;M$ NaCl in FPLC was purified with homogenity in SDS-PAGE and its enzymatic activity was confirmed in sera of invasive candidiasis with candidemia patient by immunoblotting. The purified enolase indicated no signal (100% specificity) in 40 normal human sera and 75% (6/8) sensitivity in sera of candidemic patients with suspicious invasive candidiasis by immunoblotting.

• Pediatric Infection and Vaccine
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• v.3 no.2
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• pp.139-144
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• 1996

Purpose : Coxiella burnetii is a well-known causative agent of granuolmatous inflammation and an inducer of morphological change and transformation of human B lymphocyte in vitro. Coxiella burnetii manifests with several clinical symptoms depending upon the organs that are involved. We therefore undertook to clarify the association of Kawasaki disease and Coxiella burnetii. Subjects and Methods : The patient's sera were tested for antibodies specific for Coxiella burnetii, using indirect fluorescent antibody technique(IFA). We compared Coxiella burnetii infection with 3 groups of patients, group 1 (Kawasaki disease), group 2 (other febrile disease) and group 3 (control group). Results : 1) In children with Kawasaki disease (group 1), 93% of the patient tested positive for Coxiella burnetii. 2) In group 2 children, 20% of the patient tested positive for Coxiella burnetii. 3) In group 3 children, 10% of the patient tested positive for Coxiella burnetii 4) There were significant higher positive rate for Coxiella burnetii in Kawasaki disease than group 2 and group 3 (p<0.05). Conclusions : We concluded that our cases of Kawasaki disease were associated with Coxiella burnetii infection. Further studies will be needed to understand the precise role of Coxiella infection in Kawasaki disease.

• The Journal of Korean Society of Virology
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• v.27 no.1
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• pp.49-57
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• 1997

We developed a sensitive, nested reverse transcription-polymerase chain reaction (RT-PCR) to detect Hantaan, Seoul, Belgrade, Puumala and Sin Nombre viruses in animal tissues. Total RNA was extracted from blood, lung or kidney samples of experimentally-infected hamsters by using the guanidine isothiocyanate buffer-acid phenol-chloroform method. Genus-reactive outer primers were derived from the consensus region of the G1 gene sequences of several hantaviruses. Serotype-specific primers were selected within the region amplified by the outer primers. To examine the sensitivity and specificity of the test, we diluted known quantities of Hantaan, Seoul, Belgrade, Puumala and Sin Nombre viruses in human or hamster immune sera before performing the nested RT-PCR. We could detect as little as 1 pfu of virus, even in the presence of high-titer neutralizing antibodies, and the serotype-specific primers amplified only homologous serotype viruses. RT-PCR with these primers demonstrated virus in the blood of experimentally-infected hamsters as early as four days to as late as 30 days after infection. A comparison of a standard immunofluorescent antibody screening test (IFAT) to nested RT-PCR with RNA extracted from lung or kidney tissues of the hamsters, demonstrated that RT-PCR to be more sensitive for identifying viruses in these tissues.

• Korean Journal of Veterinary Service
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• v.33 no.2
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• pp.121-128
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• 2010

Brucellosis is a serious zoonosis, recognized worldwide. It primarily affects animals, which act as reservoirs for human infection as well as being of economic significance to the agri-food industry. Bangladesh has been reported as an endemic area for brucellosis. So a cross sectional study was conducted to determine the seroprevalence and potential risk factors of brucellosis in cattle in Dinajpur and Mymensingh districts of Bangladesh. A total of 182 cattle were examined by Rose Bengal Plate Test (RBPT) between September 2008 and October 2009. Then Positive, doubtful, and negative samples were further confirmed with slow agglutination test (SAT) and both indirect and competitive enzyme linked immunosorbent assay (iELISA and cELISA). A questionnaire was used to collect epidemiological information of the animals. The overall animal-level prevalence was 3.30%. Brucellosis seroprevalence was higher (4.76% by cELISA) in cattle above 48 months than those under 48 months. Female showed higher seroprevalence (10.67%) than male (6.25%). Higher seroprevalence was also found in cattle bred naturally (20.0%) than artificially (8.77%) and cattle that aborted or with previous abortion record (22.22%) showed higher seroprevalence than non-aborted (7.69%). The sensitivity of RBT and SAT was found 100% as compared to cELISA standard test, whereas specificity of RBT (95.35%) was higher than that of SAT (94.32%).

• Bulletin of the Korean Chemical Society
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• v.34 no.2
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• pp.460-464
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• 2013

Single-chain variable fragments of antibodies (scFv) specific to 2,4-dinitrotoluene (DNT) were isolated from a phage library displaying synthetic human scFv fragments with 6 diversified complementary determining regions (CDRs). A DNT derivative that contained an extended amine group was synthesized and conjugated to the NHS-group that was linked to magnetic beads. Phages specific to the immobilized DNT derivatives were isolated from the library after 4 rounds of sequential binding and elution processes. The displayed scFv fragments from the isolated phages showed consensus CDR sequences. One DNT-specific scFv was expressed in E. coli and purified using Ni-affinity chromatography. The purified DNT-specific scFv binds specifically to the immobilized DNT-derivative with $K_D$ value of $6.0{\times}10^{-7}$ M. The scFv and DNT interaction was not disrupted by the addition of 4-nitrotoluene or benzoic acid. These data demonstrate that the screened scFv from the phage displayed library could be used for selective and sensitive detection of explosives such as TNT.

• Molecules and Cells
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• v.38 no.10
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• pp.918-924
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• 2015

During T cell activation, mitochondrial content increases to meet the high energy demand of rapid cell proliferation. With this increase, the level of reactive oxygen species (ROS) also increases and causes the rapid apoptotic death of activated cells, thereby facilitating T cell homeostasis. Nicotinamide (NAM) has previously been shown to enhance mitochondria quality and extend the replicative life span of human fibroblasts. In this study, we examined the effect of NAM on $CD8^+$ T cell activation. NAM treatment attenuated the increase of mitochondrial content and ROS in T cells activated by CD3/CD28 antibodies. This was accompanied by an accelerated and higher-level clonal expansion resulting from attenuated apoptotic death but not increased division of the activated cells. Attenuation of ROS-triggered pro-apoptotic events and upregulation of Bcl-2 expression appeared to be involved. Although cells activated in the presence of NAM exhibited compromised cytokine gene expression, our results suggest a means to augment the size of T cell expansion during activation without consuming their limited replicative potential.

• Parasites, Hosts and Diseases
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• v.40 no.4
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• pp.177-180
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• 2002

Gangweon-do is known to be highly endemic area of sparganosis more than other provinces in Korea. A seroepidmiologic examination for the detection of anti-Spirometra erinacei plerocercoid IgG in serum was carried out in normal inhabitants in Hongcheon-gun, Gangweon-do. Sere were tested by enzyme-linked immunosrobent assay (ELISA) for the anti-sparganum antibodies. Positive rate for anti-sparganum antibody in 719 adults was 3.3%. Data of the questionnaire for 24 ELISA positive inhabitants revealed that 20 had a history of eating raw meat of snakes, 24 had a history of eating frogs, and 24 had a history of drinking stream water. Two positive cases had a past history of sparganosis. Two positive cases showed current symptoms of sparganosis. The data revealed that ELISA would be useful to find infected cases among normal inhabitants at sparganosis-endemic areas.