• Proceedings of the KACD Conference
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• 2003.11a
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• pp.546.1-546
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• 2003

Laser doppler flowmeter (LDF) has been applied to the measurement of pulpal blood flow (PBF) in human teeth. As far as we searched, the detection area of the pulp in the blood flow measurement has not been clarified, yet. Therefore, the purpose of this study was to obtain information of the detection area in PBF measurement using LDF. The experiments were performed on the artificial blood circulation in extracted human upper central incisors. The apical portions of examined teeth (n=6) were severed and root canals were enlarged from the apical end to the 2mm incisal to the level of enamel-cement junction. An individual resin cap of each tooth was prepared and a hole was drilled 2mm incisal to enamel-cement junction of the labial side of the cap. The measurement probe of LDF (MBF3D, Moor Instrument, UK) was plugged into the hole of the cap. Heparinized human peripheral blood, which was in advance collected and diluted 3 times with physiological saline, was pumped through the apical foramen of the teeth via a silicone tube and a disposable needle (o.d. 0.7mm) and blood flow signals were monitored. The flux signal significantly increased with the enlargement of the root canal to incisal direction (p<0.01, Friedman analysis). The result indicates that the performance of LDF in PBF with human teeth is limited.

• Journal of Plant Biotechnology
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• v.49 no.1
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• pp.90-98
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• 2022

This study was performed to determine the optimal ratio for preparing an extract comprising the Ecklonia cava and Hizikia fusiformis complex as a therapeutic material for alleviating inflammatory respiratory diseases. First, to examine the optimal ratio for preparing the complex (SD-EH), Ecklonia cava and Hizikia fusiformis extracts were used; four extracts with different mixing ratios were prepared. The effects of the SD-EH extract on MUC5AC mRNA expression in PMA-treated NCI-H292 cells were analyzed; it was confirmed that the MUC5AC expression was significantly reduced after treatment with the SD-EHA-001 (E(100) : H(0)), SD-EHB-001 (E(90) : H(10)), SD-EHC-001 (E(80) : H(20)), and SD-EHD-001 (E(70) : H(30)) extracts. Western blotting was used to determine whether the SD-EH extract affects the expression levels of COX-2 and MMP-9 in PMA-treated A549 cells. The protein expression levels of COX-2 and MMP-9 were significantly lower (p < 0.001) in the cells treated with the SD-EHC-001 (E(80):H(20)), SD-EHD-001 (E(70) : H(30)), and SD-EHE-001 (E(60) : H(40) extracts than in the cells treated with PMA alone. The SD-EHC-001 (E(80) : H(20)) extract markedly downregulated the expression levels of MUC5AC, COX-2, and MMP-9. Therefore, the SE-EH extract may serve as a potential therapeutic agent for treating inflammatory respiratory diseases.

• Toxicological Research
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• v.17
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• pp.59-69
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• 2001

Arsenic exposure is associated with several human diseases, including cancers, atherosclerosis, hypertension, and cerebrovascular diseases. In cultured cells, arsenite, an inorganic arsenic com-pound, was demonstrated to interfere with many physiological functions, such as enhancement of oxidative stress, delay of cell cycle progression, and induction of structural and numerical changes of chromosomes. The objective of this study is to investigate the effects of arsenic exposure on gene expression profiles by colorimetric cDNA microarray technique. HFW (normal human diploid skin fibroblasts), CL3 (human lung adenocarcinoma cell line), and HaCaT (immortalized human keratinocyte cell line) were treated with 5 $\mu\textrm{M}$ or 10 $\mu\textrm{M}$ sodium arsenite for 6 or 16 h, respectively. By a dual-color detection system, the expression profile of arsenite-treated cultures was compared to that of control cultures. Several genes expressed differentially were identified on the microarray membranes. For example, MDM2, SWI/SNF, ubiquitin specific protease 4, MAP3K11, RecQ protein-like 5, and Ribosomal protein Ll0a were consistently induced in all three cell types by arsenite, whereas prohibitin, cyclin D1, nucleolar protein 1, PCNA, Nm23, and immediate early protein (ETR101) were apparently inhibited. The present results suggest that arsenite insults altered the expression of several genes participating in cellular responses to DNA damage, stress, transcription, and cell cycle arrest.

• 한국생물공학회:학술대회논문집
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• 2005.04a
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• pp.37-37
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• 2005

Human chorionic gonadotropin (hCG) is a heterodimeric glycoprotein hormone consisting of non-covalently linked two subunits, the ${\alpha}$ and ${\beta}$ subunit. It has been used as a infertility drug for ovulation to mimic luteinizing hormone $(LH).^{1)}$ A stable cell line was established by transfection of Rc/CMV-i-dhfr-hCG, expression vector containing hCG ${\alpha}-$ and ${\beta}-genes$, into dihydrofolate reductase-deficient CHO cells and subesquent methotrexate-mediated gene amplification. Anchorage-dependent CHO cells were adapted into a serum-free and/or animal component-free suspension medium through gradual serum weaning for the hCG production. The established cell line showed typical morphological characteristics and growth profile of CHO cells, and could produce FSH with passage-to-passage consistency. The high density perfusion culture of the CHO cells was carried out in Celligen Plus bioreactor equipped with a spin-filter as a internal cell retention device. The cell density reached up to $>1x10^{7}$ cells/ml in less than 7 days and a perfusion-control strategy based on cellular consumption rates of glucose was $established.^{2)}$ Biologically active recombinant hCG was purified by a series of chromatographic steps including anion exchange chromatography and hydrophobic interaction chromatography to homogeneity. The highly purified recombinant hCG was characterized for physicochemical, immunological and biological properties.

• Clinical and Experimental Pediatrics
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• v.61 no.6
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• pp.180-186
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• 2018

Purpose: Despite the availability of molecular methods, identification of the causative virus in children with acute respiratory infections (ARIs) has proven difficult as the same viruses are often detected in asymptomatic children. Methods: Multiplex reverse transcription polymerase chain reaction assays were performed to detect 15 common respiratory viruses in children under 15 years of age who were hospitalized with ARI between January 2013 and December 2015. Viral epidemiology and clinical profiles of single virus infections were evaluated. Results: Of 3,505 patients, viruses were identified in 2,424 (69.1%), with the assay revealing a single virus in 1,747 cases (49.8%). While major pathogens in single virus-positive cases differed according to age, human rhinovirus (hRV) was common in patients of all ages. Respiratory syncytial virus (RSV), influenza virus (IF), and human metapneumovirus (hMPV) were found to be seasonal pathogens, appearing from fall through winter and spring, whereas hRV and adenovirus (AdV) were detected in every season. Patients with ARIs caused by RSV and hRV were frequently afebrile and more commonly had wheezing compared with patients with other viral ARIs. Neutrophil-dominant inflammation was observed in ARIs caused by IF, AdV, and hRV, whereas lymphocyte-dominant inflammation was observed with RSV A, parainfluenza virus, and hMPV. Monocytosis was common with RSV and AdV, whereas eosinophilia was observed with hRV. Conclusion: In combination with viral identification, recognition of virus-specific clinical and laboratory patterns will expand our understanding of the epidemiology of viral ARIs and help us to establish more efficient therapeutic and preventive strategies.

• Journal of Pharmaceutical Investigation
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• v.35 no.3
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• pp.137-142
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• 2005

A rapid, selective and sensitive reversed-phase HPLC method for the determination of glipizide in human serum was validated and applied to the pharmacokinetic study of glipizide. Glipizide and internal standard, tolbutamide, were extracted from human serum by liquid-liquid extraction with benzene and analyzed on a Nova Pak $C_{18}\;60{\AA}$ column with the mobile phase of acetonitrile-potassium dihydrogen phosphate (10 mM, pH 3.5) (4:6, v/v). Detection wavelength of 275 nm and flow rate of 0.7 ml/min were fixed for the study. The assay robustness for the changes of mobile phase pH, organic solvent content, and flow rate was confirmed by $3^3$ factorial design using a fixed glipizide concentration (500 ng/ ml) with respect to its peak area and retention time. And also, the ruggedness of this method was investigated at three different laboratories using same quality control (QC) samples. This method showed linear response over the concentration range of 10-1000 ng/ml with correlation coefficient greater than 0.999. The lower limit of quantitation using 0.5 ml of serum was 10.0 ng/ml, which was sensitive enough for pharmacokinetic studies. The overall accuracy of the quality control samples ranged from 82.6 to 105.0% for glipizide with overall precision (% C.V.) being 1.13-13.20%. The percent recovery for human serum was in the range of 85.2 93.5%. Stability studies showed that glipizide was stable during storage, or during the assay procedure in human serum. The peak area and retention time of glipizide were not significantly affected by the changes of mobile phase pH, organic solvent content, and flow rate under the conditions studied. This method showed good ruggedness (within 15% C.V.) and was successfully used for the analysis of glipizide in human serum samples for the pharmacokinetic studies at three different laboratories, demonstrating the suitability of the method.

• Journal of Biosystems Engineering
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• v.34 no.2
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• pp.120-126
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• 2009

The purpose of this study was to develop a visibility evaluation system for cabin type combine. Human's field of view was classified into five levels (perceptive, effective, stable gaze, induced, and auxiliary) depending on rotation of human's head and eye. Divider, reaper lever, gearshift, dashboard, and conveying part were considered as major viewpoints of combine. Visibilities of combine was evaluated quantitatively using the viewpoints and the human's field of view levels. The visibility evaluation system for cabin type combine was consisted of a laser pointer, stepping motors to control the direction of view, gyro sensors to measure horizontal and vertical angle, and I/O interface to acquire the signals. Tests were conducted with different postures ('sitting straight', 'sitting with $15^{\circ}$ tilt', 'standing straight', and 'standing with $15^{\circ}$ tilt'). The LSD (least significant difference) multiple comparison tests showed that the visibilities of viewpoints were different significantly as the operator's postures were changed. The results showed that the posture at standing with $15^{\circ}$ tilt provided the best visibility for operators. The divider of the combine was invisible due to blocking with the cabin frame at many postures. The reaper lever showed good visibilities at the postures of sitting or standing with $15^{\circ}$ tilt. The gearshift, the dashboard, and the conveying part had reasonable visibilities at the posture of sitting with $15^{\circ}$ tilt. However, most viewpoints of the combine were out of the stable gaze field of view level. Modifications of the combine design will be required to enhance the visibility during harvesting operation for farmers' safety and convenience.

• Molecules and Cells
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• v.41 no.7
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• pp.684-694
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• 2018

Upregulation of human telomerase reverse transcriptase (hTERT) expression is an important factor in the cellular survival and cancer. Although growing evidence suggests that hTERT inhibits cellular apoptosis by telomere-independent functions, the mechanisms involved are not fully understood. Here, we show that hTERT contains a BH3-like motif, a short peptide sequence found in BCL-2 family proteins, and interacts with anti-apoptotic BCL-2 family proteins MCL-1 and BCL-xL, suggesting a functional link between hTERT and the mitochondrial pathway of apoptosis. Additionally, we propose that hTERT can be categorized into the atypical BH3-only proteins that promote cellular survival, possibly due to the non-canonical interaction between hTERT and antiapoptotic proteins. Although the detailed mechanisms underlying the hTERT BH3-like motif functions and interactions between hTERT and BCL-2 family proteins have not been elucidated, this work proposes a possible connection between hTERT and BCL-2 family members and reconsiders the role of the BH3-like motif as an interaction motif.

• IMMUNE NETWORK
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• v.2 no.2
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• pp.91-95
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• 2002

Background: In order to characterize human antibodies with specificity for mite allergens at the molecular level, a scFv phage display library was constructed using peripheral blood mononuclear lymphocytes from an asthma patient allergic to mite as Ig gene sources. Methods: Immunoglobulin $V_H$ and V gene fragments were obtained by polymerase chain reaction, and randomly combined in pCANTAB-5E vector. The resulting human scFv phage display library had $3{\times}10^4$ independent clones, and biopanning was performed with house dust mite extracts. Results: Four scFv clones specific for house dust mite extract were isolated. Immunoblot assay showed that our clones reacted to 25 kDa and 50~60 kDa proteins with unknown identity in mite extracts. Sequence analysis indicated that two clones (b7 and c15) are identical, and all clones belong to human $V_H3$ subgroup. On the other hand, light chain usage was different in that two clones (a2 and b7 / c15) belonging to V ${\kappa}4$ subgroup, but a4 used V ${\kappa}1$ light chain gene. Conclusion: Our approach should facilitate provision of useful information on the antibody responses against allergens at the molecular level in humans.

• KSBB Journal
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• v.15 no.3
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• pp.238-242
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• 2000

A fusion protein of human interleukin-2(hiL-2) and glucagon which was expressed in Escherichia coli. was digested with enterokinase for recovery of hIL-2 from the fused protein. To obtain hIL-2 of optimum recovery hydrolysis reaction were performed under various conditions of urea additives and reaction time. hIL-2 was finally purified by RP-HPLC(reversed phase-HPLC) to remove cleaved G3 fusion partner and residual uncleaved G3-IL-2 HIL-2 was eluted in a single peak at 100% acetonitrile at 28 min. Optimum urea concentration was found to be 0.5 M and 24 h reaction time was sufficient without any additive such as CaCl2 and Tween-20.