• Development and Reproduction
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• 제6권1호
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• pp.55-66
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• 2002

Embryonic stem cells(ES cells) are derived from the inner cell mass(ICM) of blastocysts, which have the potentials to remain undifferentiated, to proliferate indefinitely in vitro, to differentiate into the derivates of three embryonic germ layers. ES cells are an attractive model system for studying the initial developmental decisions and their molecular mechanisms during embryogenesis. Additionally, ES cells of significant interest to those characterizing the various gene functions utilizing transgenic and gene targeting techniques. We investigated the effects of reproductive hormones, gonadotropins(GTH) and steroids on the induction of differentiation and expressions of their receptor genes using the newly established mouse ES cells. We collected the matured blastocysts of inbred mice C57BL/6J after superovulation and co-cultured with mitotically inactivated STO feeder cells. After 5 passages, we confirmed the expression alkaline phosphatase(Alk P) activity and SSEA-1, 3, 4 expressions. The protocol devised for inducing ES differentiation consisted of an aggregation steps, after 5 days as EBs in hormone treatments(FSH, LH, E$_2$, P$_4$, T) that allows complex signaling to occur between the cells and a dissociation step, induced differentiation through attachment culture during 7 days in hormone treatments. Hormone receptors were not increased in dose-dependent manner. All hormone receptors in ES cells treated reproductive hormones were expressed lower than those of undifferentiated ES cell except for LHR expression in E$_2$-treated ES cells group. After hormone induced differentiation, at least some of the cells are not terminally differentiated, as is evident from the expression of Oct-4, a marker of undifferentiated. To assess their differentiation by gene expression, we analyzed the expression of 7 tissue-specific markers from all three germ layers. Most of hormone-treated group increased in the expression of gata-4 and $\alpha$ -fetoprotein, suggesting reproductive hormone allowed or induced differentiation of endoderm.

• Tuberculosis and Respiratory Diseases
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• 제39권1호
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• pp.62-72
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• 1992

Background: T-cell mediated cellular immunity has been suggested as an important mechanism in mycobacterial infection and imbalance between helper/inducer and suppressor/cytotoxic T-cell has been suggested as an important immunological abnormality in the pathogenesis of tuberculosis in human. Method: To determine whether there is any difference in T-cell mediated immunity in the pathogenesis of pulmonary and extra pulmonary tuberculosis, total numbers of WBC&lymphocytes were counted and helper/inducer and suppressor/cytotoxic cells were calculated by flow cytometry. Blastogenesis after stimulation with Concanavalin-A, Phytohemagglutinin and PPD were measured by $^3H$-thymidine uptake. PPD skin test was performed as an in vivo test. Results: 1)There was no significant difference in the size of PPD skin test between pulmonary and extrapulmonary tuberculosis groups. 2)Number of total lymphocytes significantly decreased in tuberculosis patients compared with healthy control group. But there was no significant difference between pulmonary and extrapulmonary tuberculosis groups. 3) Number of HLA-DR and Interleukin-2 receptor (+) cells were significantly increased in tuberculosis patients. But there was no significant difference between pulmonary and extra pulmonary tuberculosis groups. 4) There was no significant difference in the numbers of WBC, $T_3$, $T_4$ and $T_8$ lymphocytes and $T_4/T_8$ ratio between tuberculosis patients and healthy controls. 5) There was no significant difference in the blastogenesis after stimulation with specific and non-specific blastogens between tuberculosis patients and healthy controls. 6) The percentage and absolute number of $T_4$ lymphocyte were significantly correlated with the size of PPD skin test. (r=0.689 and 0.598). Conclusion: From these results, it is concluded that there was no difference in T-cell mediated immunity between pulmonary and extra pulmonary tuberculosis group. But, because it is suspected that there might be some difference in the role of T-cell mediated immunity in the pathogenesis of pulmonary and extra pulmonary tuberculosis or even among the extrapulmonary tuberculosis patients, further studies would be required.

• Tuberculosis and Respiratory Diseases
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• 제40권6호
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• pp.683-693
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• 1993

Background: During movement the major inputs to nervous system come from firstly the muscle and joint to maintain posture and motion and secondly the chemoreceptors and baroreceptors to adjust the cardiovascular and respiratory function. Their complex relationships are generally studied for many years but the direct relation between the joint and respiratory system is not studied thoroughly until now. So this experiment was performed to determine whether the natural movement of knee joint can cause the enhancement of respiratory function by observation of the changes of respiratory rate, phrenic nerve activity and inspiratory neuron activity during the stimulation of knee joint in cat anesthetized with $\alpha$-chloralose. Method: Twenty six male adult cats were used and the extracelluar recording using bipolar platinum electrode and carbon filament electrode was done to record the changes in the activities of phrenic nerve and inspiratory neuron movement of knee joint, injection of chemicals into the joint cavity and electrical stimulation of articular nerve were done. Results: The 60 Hz. could not but 120 Hz. flexion-extension movement of knee joint increased respiratory rate(R.R.), tidal neural activity(TNA) and minute neural activity(MNA). Intra-articular injection of lactate could not increase R.R. but significantly increase TNA and MNA which represented the enhanced respiratory function. Injection of potassium chloride showed similar effects with the case of lactate but the duration of effect was shorter. The electrical stimulation of medial articular nerve with IV strength which could activate only group I and II afferents showed increased TNA and MNA during stimulation but 20 V stimulation which could activate all the afferents increased all the respiratory parameters. The changes of inspiratory neuron activity by knee joint stimulation was similar to that of phrenic nerve. Conclusion: The respiratory center could be directly stimulated by the activation of group I and II articular afferents and it seemed that the magnitude of the respiratory center enhancement is proportional to the amount of sensory information from the knee joint. These facts might suggest that the respiratory function could be enhanced even by the normal movement of knee joint.

• Tuberculosis and Respiratory Diseases
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• 제44권3호
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• pp.601-610
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• 1997

Background : Monocytes/macrophages play a central role in determining the host response during Gram-negative infection through secretion of a variety of mediators after stimulation of LPS. Even though cytokine production has been shown to play an important role in host defense during sepsis, cytokine release may also lead to tissue injury. Thus, regulation of macrophage response to LPS is critical for host survival during Gram-negative sepsis. In animals exposed to nonlethal doses of endotoxin, a characteristic hyporesponsiveness to subsequent administration of endotoxin has been observed. This phenomenon was known as 'LPS tolerance'. However, little information is available regarding the underlying mechanism of LPS tolerance. Method : Peripheral blood monocyte(PBMC) was isolated from peripheral blood of normal volunteers by adhesion purification method. To evaluate the conditions to obtain LPS tolerance, preculture was carried out with LPS at 10ng/ml for 24 hours. For stimulation, culture plates were washed two times and were stimulated with LPS at $1{\mu}g/ml$ for 4, 6 and 26 hours. To assess the underlying mechanisms of LPS tolerance, autologous serum, PMA, anti-CD14 Ab, Indomethacin or $PGF_2$ were added to preculture solution respectively. Cytokine concentrations in culture supernatants were measured using ELISA for TNF-$\alpha$ and IL-8 and mRNA of TNF-$\alpha$ and IL-8 were determined by Northern blot analysis. Results : The exposure of PBMC to low dose of LPS suppressed the cytokine production and mRNA expression of TNF-$\alpha$, but not IL-8. Anti-CD14 Ab partially recovered production of TNF-$\alpha$ which was suppressed by preculture with low dose LPS. The preculture with PMA induces LPS tolerance, as preculture with low dose LPS. Conclusion : LPS tolerance to TNF-$\alpha$ is regulated pretranslationally and is influenced by protein kinase C pathway and CD14.

• Korean Journal of Microbiology
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• 제38권3호
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• pp.153-161
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• 2002

Sox4, a transcription factor, consists of three functional domains: an HMG-box domain as a DNA binding domain, serine rich region as a transactivation domain and glycine rich region (GRR), an unknown functional domain. Although Sox4 is known to be functionally involved in heart, B-cell and reproductive system development, its physiological function remains to be elucidated. We used pGEX expression system to develop a simple and rapid method for purifying Sox4 protein in suitable forms for biochemical studies of their functions. Unexpectedly, we observed that full-length Sox4 appears to be protease-sensitive during expression and purification in E. coli. To map the protease-sensitive site in Sox4, we generated various constructs with each of functional domains of Sox4 and purified as the GST-Sox4 fusion proteins using glutathione beads. We found that the specific cleavage site for the proteolytic enzyme, which exists in E. coli, is localized within the novel GRR of Sox4. Our study suggest that the GRR of Sox4 may a target for the cellular protease action and this cleavage in the GRR may be involved in regulating physiological function of Sox4. Additionally, our study may provide a useful method for investigating the proteolytic cleavage of the target molecule in E. coli.

• Clinical and Experimental Pediatrics
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• 제51권1호
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• pp.54-61
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• 2008

Purpose : This study was designed to clarify the mechanism of proteinuria in nephrotic syndrome patients by using puromycin aminonucleoside (PAN) nephrosis model. Methods : Following administration of various concentrations of PAN and antioxidants we observed the changes of podocyte cytoskeletons in cultured rat glomerular epithelial cells (GEpC) by method of scanning electron microscope, reactive oxyten species (ROS) analysis, permeability assay, confocal microscope, and Western blot assay. Results : PAN not only induced the ultrastructural changes of GEpC, such as shortening and fusion of microvilli, but also separated the intercellular gaps and linear ZO-1. PAN induced oxidative stresses in time and dose dependent manners and increases of intercellular permeability in anti-oxidants inhibitable manners. High concentration of PAN induced not only actin polymerization and disorganization, but also the conglomerulation and internal dislocation of ${\alpha}-actinin$ protein. The intensities of fluorescences of ZO-1 protein were diminished and internalized by PAN in a dose-dependent manner, which were inhibited by anti anti-oxidants. Conclusion : PAN induced the changes of podocytes cytoskeleton and junctional barriers by way of increasing ROS in GEpC that resulted in increasing their permeability in a antioxidatn-inhibitable manner. Glomerular hyperpermeability induced by PAN mediateing through oxidative stresses is thought to take part in the mechanism of proteinuria in nephrotic syndrome.

• Korean Journal of Plant Resources
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• 제26권2호
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• pp.149-158
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• 2013

The objective of this study was to investigate the antioxidative capacity of ethanol extracts from Sanguisorbae officinalis L. root (Sanguisorbae radix) in vitro. The concentration of Sanguisorbae radix extract at which DPPH radical scavenging activity was inhibited by 50% was 0.33 mg/mL, which was similar to $IC_{50}$ of ${\alpha}$-tocopherol (0.40 mg/mL), as compared to 100% by pyrogallol as a reference. Total antioxidant status was examined by total antioxidant capacity against ABTS radical reactions. Total antioxidant capacities of Sanguisorbae radix extract were significantly (p<0.05) higher than those of ${\alpha}$-tocopherol. Superoxide scavenging activities of Sanguisorbae radix extract were significantly (p<0.05) higher than those of catechin. Oxygen radical absorbance capacities of Sanguisorbae radix extract were significantly (p<0.05) higher than those of ascorbic acid. Cupric reducing antioxidant capacities of Sanguisorbae radix extract were significantly (p<0.05) higher than those of ${\alpha}$-tocopherol. Sanguisorbae radix extract prevented supercoiled DNA strand breakage induced by hydroxyl radical and peroxyl radical. Total phenolic contents of Sanguisorbae radix extract at concentrations of 0.5 and 5 mg/mL were 0.50 and 3.33 mM gallic acid equivalents, respectively. Sanguisorbae radix extract at concentration of 0.01, 0.1 and 0.5 mg/mL inhibited 0.2 mM tert-butyl hydroperoxide-induced cytotoxicity by 33.8, 79.1 and 96.9%, respectively, in HepG2 cell culture system. Thus, strong antioxidant and cytotoxicity-ihnibiting effects of Sanguisorbae radix extract seem to be due to, at least in part, the prevention from free radicals-induced oxidation as well as high levels in total phenolic contents.

• Polymer(Korea)
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• 제29권3호
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• pp.260-265
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• 2005

The initial burst of drug release is an important role in the controlled delivery of drug having hish toxicity and narrow therapeutic ranges. Nanosphere composed of monolayer could not achieve precisely controlled drug release because of the initial burst of drug on surface. In this study, double layered nanosphere was prepared for sustained drug delivery without initial burst. Double layered nanosphere composed of dextran and PLGA was fabricated by using conventional W/O/W double emulsion method. To control surface tension on the outer layer of nanospheres, PVA was used as a surfactant. Release behavior of dextran as model drug was observed as the $3{\times}1$mm wafers formed by compression mould in the deionized water for 7 days. Double layered nanosphere has sustained release behavior, in contast to single layered nanospheres. such as mechanical mixture and dextran nanospheres. Especially, nanosphere containing PVA $0.2\%$ has shown nearly the zero-order release profile. As a result of this study, double layered nanospheres has more sustained release profile of drug without the initial burst and the release behavior of dexoan on tile double layered nanospheres was controlled by the contents of PVA as a surfactant.

• Korean Journal of Optics and Photonics
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• 제30권6호
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• pp.243-248
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• 2019

The respiratory tract is an essential part of the respiratory system involved in the process of respiration. However, if stenosis occurs, it interferes with breathing and can even lead to death. Asthma is a typical example of a reversible cause of airway narrowing, and the number of patients suffering from acute exacerbation is steadily increasing. Therefore, it is important to detect airway narrowing early and prevent the patient's condition from worsening. Optical coherence tomography (OCT), which has high resolution, is suitable for observing the microstructure of tissues. In this study we developed an endoscopic OCT system. We combined a 1300-nm OCT system with a servo motor, which can rotate at a high speed. A catheter was pulled back using a linear stage while imaging with 360° rotation by the motor. The motor was selected considering various requirements, such as torque, rotational speed, and gear ratio of pulleys. An ex vivo rabbit tracheal model was used as a sample, and the sample and catheter were immobilized by acrylic structures. The OCT images provided information about the structures of the mucosa and submucosa. The difference between normal and stenosed parts in the trachea was confirmed by OCT. Furthermore, through a three-dimensional (3-D) reconstruction process, it was possible to identify and diagnose the stenosis in the 3-D image of the airway, as well as the cross-sectional image. This study would be useful not only for diagnosing airway stenosis, but also for realizing 3-D imaging.

• Journal of the korean academy of Pediatric Dentistry
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• 제29권1호
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• pp.76-83
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• 2002

The purpose of study was to determine the effectiveness of crude IgY to S. mutans in preventing the acid production and the demineralization of primary tooth enamel in vitro. The acid production by S. mutans in Todd Hewitt broth with and without 5% sucrose was inhibited by 2.5% crude IgY, and as the concentration of crude IgY increased from 2.5% to 17.5%, the pH drop of the media after incubation continued to decrease. There were high positive correlations between the concentration of crude IgY and the pH of media in the late incubation period. The inhibition rate of demineralization of primary tooth enamel by S. mutans was determined by measuring the surface microhardness after incubation in 5% sucrose Todd Hewitt broth for 12 hours. The inhibition rate was 32.28% in 2.5% IgY, 42.28% in 7.5% IgY, 64.06% in 12.5% IgY, and 92.79% in 17.5% IgY. There was high positive correlation between the concentration of crude IgY and the surface microhardness of enamel after demineralization These results suggest that it would be possible to prevent dental caries through passive immunization using crude IgY.