• Tuberculosis and Respiratory Diseases
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• v.73 no.5
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• pp.258-265
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• 2012

Background: Vitamin D can translocate a vitamin D receptor (VDR) from the nucleus to the cell membranes. The meaning of this translocation is not elucidated in terms of a role in pathogenesis of chronic obstructive pulmonary disease (COPD) till now. VDR deficient mice are prone to develop emphysema, suggesting that abnormal function of VDR might influence a generation of COPD. The blood levels of vitamin D have known to be well correlated with that of lung function in patients with COPD, and smoking is the most important risk factor in development of COPD. This study was performed to investigate whether cigarette smoke extracts (CSE) can inhibit the translocation of VDR and whether mitogen activated protein kinases (MAPKs) are involved in this inhibition. Methods: Human alveolar basal epithelial cell line (A549) was used in this study. 1,25-$(OH_2)D_3$ and/or MAPKs inhibitors and antioxidants were pre-incubated before stimulation with 10% CSE, and then nucleus and microsomal proteins were extracted for a Western blot of VDR. Results: Five minutes treatment of 1,25-(OH2)D3 induced translocation of VDR from nucleus to microsomes by a dose-dependent manner. CSE inhibited 1,25-$(OH_2)D_3$-induced translocation of VDR in both concentrations of 10% and 20%. All MAPKs inhibitors did not suppress the inhibitory effects of CSE on the 1,25-$(OH_2)D_3$-induced translocation of VDR. Quercetin suppressed the inhibitory effects of CSE on the 1,25-$(OH_2)D_3$-induced translocation of VDR, but not in n-acetylcysteine. Conclusion: CSE has an ability to inhibit vitamin D-induced VDR translocation, but MAPKs are not involved in this inhibition.

• Biomolecules & Therapeutics
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• v.32 no.1
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• pp.56-64
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• 2024

Biased signaling or functional selectivity refers to the ability of an agonist or receptor to selectively activate a subset of transducers such as G protein and arrestin in the case of G protein-coupled receptors (GPCRs). Although signaling through arrestin has been reported from various GPCRs, only a few studies have examined side-by-side how it differs from signaling via G protein. In this study, two signaling pathways were compared using dopamine D₂ receptor (D₂R) mutants engineered via the evolutionary tracer method to selectively transduce signals through G protein or arrestin (D₂G and D₂Arr, respectively). D₂G mediated the inhibition of cAMP production and ERK activation in the cytoplasm. D₂Arr, in contrast, mediated receptor endocytosis accompanied by arrestin ubiquitination and ERK activation in the nucleus as well as in the cytoplasm. D₂Arr-mediated ERK activation occurred in a manner dependent on arrestin3 but not arrestin2, accompanied by the nuclear translocation of arrestin3 via importin1. D₂R-mediated ERK activation, which occurred in both the cytosol and nucleus, was limited to the cytosol when cellular arrestin3 was depleted. This finding supports the results obtained with D₂Arr and D₂G. Taken together, these observations indicate that biased signal transduction pathways activate distinct downstream mechanisms and that the subcellular regions in which they occur could be different when the same effectors are involved. These findings broaden our understanding on the relation between biased receptors and the corresponding downstream signaling, which is critical for elucidating the functional roles of biased pathways.

• Proceedings of the Korean Society of Applied Pharmacology
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• 1994.04a
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• pp.231-231
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• 1994

이 연구는 인삼 성분 약물이 도파민 수용체에 미치는 영향을 연구하기 위한 첫 단계로써 그 기본적인 assay system을 정착 시키기 위함이 목적이었다. 우리는 이 연구를 수행하기 위하여 생후 2-4 일의 쥐를 사용하여 흑질, 선조체, 해마구, nucleus accumbens등의 뇌 부위에서 신경세포 배양을 시도하였다. 흑질로 부터 배양한 신경세포들의 경우엔 면역 세포학적방법을 써서 살펴본 결과 대부분의 신경이 도파민성 신경이나 GABA성 신경들 이었다. 또한 이들 세포들의 전기 생리학적 상태를 알아보기 위하여 흑질에서 신호 전달체계가 잘 확립된 GABA 수용체의 작용을 살펴 본 결과 이 신경세포들은 GABA-A 및 GABA-B 수용체의 발현은 물론 이온 채널에 미치는 신호 전달체계를 완전히 갖추고 있었다. 도파민 수용체의 작용을 전기 생리학적으로 연구하기 위하여 배양한 신경세포에 도파민agonist를 가해서 이온 채널에 미치는 효과를 살펴 보았다. 선조체에서 배양한 신경세포들은 Dl 과 D2 agonist에 대해서 상반되는 반응을 나타냈다. 즉 Dl agonist는 선조체 신경세포를 활성화 시켰으나 D2 agonist는 선조체 신경세포들을 억제 하였다. 한편 해마구의 CAI 과 CA3 부위로 부터 배양한 신경세포에 대한 도파민 agonists의 작용은 선조체의 신경세포에 대한도파민 agonists의 작용과는 상반되는 반응 이었다. Dl agonist는 해마구로 부터 배양한 신경세포의 활성을 억제 하였으나 D2 agonist는 이들 신경세포들의 활성을 증가 시켰다. Nucleus accumbens 에서 배양한 신경세포들은 도파민에 의해서 그 활성이 억제 되었다. 이러한 결과로 미루어 봐서 같은 도파민 수용체라도 분포되어 있는 조직에 따라서 신호전달 에 관여 하고 있는 C-단백이나 이차 전령물질이 달라서 신경세포에 대한 작용이 다르든지. 약리학적으로는 구분되지 않으나 뇌의 조직에 따라서 분포가 다른 도파민 수용체의 아그룹이 존재 한다고 생각된다.

• Journal of Magnetics
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• v.16 no.3
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• pp.308-311
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• 2011

In order to obtain mono-dispersed Fe NPs with high saturation magnetization, quantitative analysis method to investigate the growth dynamics of the Fe NPs synthesized by a conventional thermal decomposition method has been developed. As a result, fast nucleation process promotes formation of ~4 nm of initial nucleus with a non-equilibrium phase, resulting in low saturation magnetization. And slow particle growth with atomic-scaled surface precipitation mode (< 100 atoms/($min{\cdot}nm^2$)) can form the growth layer on the surface of initial nucleus with high saturation magnetization (~190 emu/$g_{Fe}$) as an equilibrium a phase of Fe. Therefore, higher stabilization of small initial nucleus generated just after the injection of $Fe(CO)_5$ should be one of the key issues to achieve much higher $M_s$ of Fe NPs.

• The Journal of Korean Physical Therapy
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• v.9 no.1
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• pp.59-70
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• 1997

The ability of neurotropic alpha herpesviruses to replicate within synaptically linked neurons has made these pathogens valuable tools for transneuronal analysis. Recent studies suggest that unique gene products expressed by genetically engineered strains of virus may permit the use of multiple strains in complex tracing paradigms. In the present study we have examined the invasiveness of two genetically engineered strains of the swine pathogen known as pseudorabies virus(PRV). The two strains were isogenic with the attenuated Bartha strain of PRV; in one strain a lacZ reporter gene was inserted into the gC locus (PRV-BaBlu; $4.75\times10^8pfu/ml$) contrained a PRV envelope glycoprotein gene that was absent in PRV-BaBlu. Simultaneous or temporally separated sequential injection of $4\mu\ell$ of each strain into the ventral wall of the stomach produced a predictale course of retrograde synaptic infection. The results were as follows: 1. PRV-BaBlu and PRV-D infected the dorsal motor nucleus of vagus nerve(DMV) and paraventricular nucleus(PVN). 2. Invasion and replication of PRV-D occured at a faster rate than the parental strain or PRV-BaBlu. 3. PRV-D was much more virulent than PRV-BaBlu or the parental strain. 4. Co-injection of PRV-D and PRV-BaBlu produced an infection that was more virulent than that produced by the parental strain (PRV-Bartha), 5. Neurons in DMV were permissive to co-infection with PRV-D and PRV-BaBlu when they were injected simultaneously into the same site. 6. Replication of PRV-BaBlu was compromised by prior infection of the same circuit with PRV-D. 7. Prior infection of neurons with PRV-D maked them resistant to infection with PRV-BaBlu.

• Journal of Biomedical Engineering Research
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• v.39 no.1
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• pp.22-29
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• 2018

Intervertebral disc(IVD) mainly consists of Annulus fibrosus(AF) and Nucleus pulposus(NP), playing a role of distributing a mechanical load on vertebral body. IVD tissue engineering has been developed the methods to achieve anatomic morphology and restoration of biological function. The goal of present study is to identify the possibilities for creating a substitute of IVD the morphology and biological functions are the same as undamaged complete IVD. To fabricate the AF and NP combine biphasic IVD tissue, AF tissue scaffolds have been printed by 3D bio-printing system with natural biomaterials and NP tissues have been prepared by scaffold-free culture system. We evaluated whether the combined structure of 3D printed AF scaffold and scaffold-free NP tissue construct could support the architecture and cell functions as IVD tissue. 3D printed AF scaffolds were printed with 60 degree angle stripe patterned lamella structure(the inner-diameter is 5mm, outer-diameter is 10 mm and height is 3 mm). In the cytotoxicity test, the 3D printed AF scaffold showed good cell compatibility. The results of histological and immunohistochemical staining also showed the newly synthesized collagens and glycosaminoglycans, which are specific makers of AF tissue. And scaffold-free NP tissue actively synthesized glycosaminoglycans and type 2 collagen, which are the major components of NP tissue. When we combined two engineered tissues to realize the IVD, combined biphasic tissues showed a good integration between the two tissues. In conclusion, this study describes the fabrication of Engineered biphasic IVD tissue by using enable techniques of tissue engineering. This fabricated biphasic tissue would be used as a model system for the study of the native IVD tissue. In the future, it may have the potential to replace the damaged IVD in the future.

• Journal of Life Science
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• v.20 no.1
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• pp.1-8
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• 2010

We recently reported the distributions of AMPA ($\alpha$-amino-3-hydroxyl-5-methyl-4-isoxazole-propionate) receptor subtypes glutamate receptors (GluR) 1 and GluR4 in the superior colliculi (SC) of hamsters with antibody immunocytochemistry and the effect of enucleation on these distributions. We also compared these labelings to those of calcium-binding proteins calbindin D28K, calretinin, parvalbumin, and GABA. In the present study, we investigated whether the GluR1- and GluR4-immunoreactive (IR) neurons are interneurons or projection neurons by injection of the retrograde tracer horseradish peroxidase (HRP) into one of each major ascending and descending pathways of the SC. HRP injections were made into a tecto-reticulospinal pathway (TRS) and dorsal lateral geniculate nucleus (dLGN). Animals were then allowed to recover and to survive for 48 hr before perfusion. Sections containing retrograde-labeled neurons were then treated for GluR-immunoreactivity. HRP injections proved that only a small population of the GluR1-IR cells project into TRS (1.4%) and dLGN (2.6%). However, a large subpopulation of GluR4-IR cells project into TRS (32.7%). The differential compositions of inter/projection neurons, along with our previous studies on the separate distribution of the GluR subunits, its differential co-localization with calcium-binding proteins and GABA, and differential reactions to enucleations, strongly imply the functional variety of the receptor subunits in visual behavior responses.

• Journal of Sericultural and Entomological Science
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• v.13 no.1
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• pp.31-34
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• 1971

The pupa (7th. day after mounting) were irradiated for foully minutes with 1,000${\gamma}$-ray and the copulation was done under the various conditions when the pupa grew to be imagos. The silkworm eggs that the imagos laid were divided into many groups by five, fifteen, foully-five, ninety, seven-hundred, twenty and twenty-four hundred minutes eggs respectively and the lipid phosphorus in the each group of eggs was analyzed as follows. 1. The quantity of 928${\gamma}$/100mg dry powder (D. P.) was shown in the group of the fourt-five minutes eggs whose ovum nucleus and sperm nucleus were not conjugate in the control C (male and female). The number of the greatest quantity of 944${\gamma}$/100mg D.P. after the syngamy of ovum nucleus and sperm nucleus while the smallest quantity was 768${\gamma}$/100mg D.P. in the group of ninety minutes eggs. 2. Not much difference could be seen the copulation group of female irradiation and male control and the copulation group of male irradiation and female control compared to the both male and female control group, but the influence could be seen in the group of irradiated female pupas. 3. The great difference could be seen in the group of irradiated male and female pupa compared to the control group. The five minutes eggs showed the smallest quantity of 536${\gamma}$/100mg D.P. and the quantity of lipid phosphorus rapidly began to increase from the fifteen minutes eggs. The quantity amounted to 1,082${\gamma}$/100 mg D.P. in the case of seven hundred and twenty minutes eggs while the quantity decreased in the twenty-four hundred minutes eggs which showed 912${\gamma}$/100 mg D.P. The general result obtained in the present study was that the irradiated male and female pupa were greatly influenced before and after the syngamy of gamete.

• Applied Microscopy
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• v.26 no.3
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• pp.305-313
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• 1996

Acinous gland cells (A, B, C, D and E-type cells) and duct cell (G-type cell) are observed in acinus and in duct of salivary gland of Korean Slug respectively by electron microscope. The type-A gland cells are numerous and are packed with medium electron dense granules (diameter, $3{\mu}m$) in cytoplasm. The circular shaped nucleolus and evenly developed chromatins are observed in the nucleus of type-B cell, and cytoplasm includes medium electron dense granules (diameter, $2.5X3.7{\mu}m$). The type-C gland cell has a round nucleus, and thin elongated-shaped heterochromatins are evenly distributed in the nucleoplasm and many net shaped endoplasmic reticulums and oval serous granules of middle electron density (diameter, $3.5X5{\mu}m$) fill the cytoplasm. The type-D gland cell is the largest and the most numerous of the gland cells consisting the salivary gland and heterochromatins in nucleus are well developed in the nucleoplasm. Most of granules (diameter, $0.8X2.5{\mu}m$) in cytoplasm are round, and look dark for the high electron density, and cytoplasm is filled with net-shaped endoplasmic reticulums. The type-E gland cells are rarely existent around the salivary gland, and the granules of those cells are irregular in shape and size and are vacuolized in cytoplasm. Intralobular salivary duct is composed of the high electron dense squamus endotheliums, while the other interlobular salivary duct is filled with irregular columnar epitheliums. The interlobular duct cell contains the high electron dense granules (size, $0.3{\sim}1.5{\mu}m$) in cytoplasm and those granules are secreted into cilia of salivary lumen.

• Proceedings of the Korean Society of Crop Science Conference
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• 2017.06a
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• pp.149-149
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• 2017

The RING (Really Interesting New Gene) finger proteins are known to play crucial roles in various abiotic stresses in plants. In this study, we report on RING finger E3 ligase, ${\underline{O}ryza}$ ${\underline{s}ativa}$ ${\underline{s}alt$-, ${\underline{A}BA}$- and ${\underline{d}rounght}$ stress-${\underline{i}nduced}$ RING finger ${\underline{p}}rotein{\underline{1}}$ gene (OsSAD1). In vitro ubiquitination assay demonstrated that unlike OsSAD1, a single amino acid substitution ($OsSAD1^{C168A}$) of the RING domain showed no E3 ligase activity, supporting the notion that the activity of most E3s is specified by a RING domain. Result of Yeast-Two hybridization, In vivo protein degradation assay supports that OsSAD1 interacting with 3 substrate, OsSNAC2, OsGRAS44 and OsPIRIN1, and mediates proteolysis of 3 substrates via the 26S proteasome pathway. Subcellular localizations of OsSAD1 while approximately 62% of transient signals were detected in cytosol, 38% of signals were showed nucleus. However, transiently expression of OsSAD1 was detected in cytosol 30% while as 70% of nucleus under 200 mM salt treated rice protoplasts. Results of bimolecular fluorescence complementation (BiFC) showed that two nucleus-localized proteins (OsSNAC2 and OsGRAS44) interacted with OsSAD1 in the both cytosol and nucleus. Heterogeneous overexpression of OsSAD1 Heterogeneous overexpresssion of OsSAD1 in Arabidopsis exhibited sensitive phenotypes with respect to Salt-, mannitol-responsive seed germination, seedling growth. In ABA conditions, OsSAD1 overexpression plants showed highly tolerance phenotypes, such as root length and stomatal closure. Our findings suggest that the OsSAD1 may play a negative regulator in salt stress response by modulating levels of its target proteins.