• Clinical and Experimental Pediatrics
• /
• v.52 no.5
• /
• pp.594-602
• /
• 2009

Purpose : Transforming growth factor (TGF)-${\beta}1$ reportedly increases neuronal survival by inhibiting the induction of inducible nitric oxide synthase (NOS) in astrocytes and protecting neurons after excitotoxic injury. However, the neuroprotective mechanism of $TGF-{\beta}1$ on hypoxic-ischemic (HI) brain injury in neonatal rats is not clear. The aim of this study was to determine whether $TGF-{\beta}1$ has neuroprotective effects via a NO-mediated mechanism and N-methyl-D-aspartate (NMDA) receptor modulation on perinatal HI brain injury. Methods : Cortical cells were cultured using 19-day-pregnant Sprague-Dawley (SD) rats treated with $TGF-{\beta}1$ (1, 5, or 10 ng/mL) and incubated in a 1% O2 incubator for hypoxia. Seven-day-old SD rat pups were subjected to left carotid occlusion followed by 2 h of hypoxic exposure (7.5% $O_2$). $TGF-{\beta}1$ (0.5 ng/kg) was administered intracerebrally to the rats 30 min before HI brain injury. The expressions of NOS and NMDA receptors were measured. Results : In the in vitro model, the expressions of endothelial NOS (eNOS) and neuronal NOS (nNOS) increased in the hypoxic group and decreased in the 1 ng/mL $TGF-{\beta}1-treated$ group. In the in vivo model, the expression of inducible NOS (iNOS) decreased in the hypoxia group and increased in the $TGF-{\beta}1$-treated group. The expressions of eNOS and nNOS were reversed compared with the expression of iNOS. The expressions of all NMDA receptor subunits decreased in hypoxia group and increased in the $TGF-{\beta}1$-treated group except NR2C. Conclusion : The administration of $TGF-{\beta}1$ could significantly protect against perinatal HI brain injury via some parts of the NO-mediated or excitotoxic mechanism.

• Proceedings of the Korean Institute of Surface Engineering Conference
• /
• 2009.05a
• /
• pp.111-112
• /
• 2009

Diamond-like carbon (DLC) is a convenient term to indicate the compositions of the various forms of amorphous carbon (a-C), tetrahedral amorphous carbon (ta-C), hydrogenated amorphous carbon and tetrahedral amorphous carbon (a-C:H and ta-C:H). The a-C film with disordered graphitic ordering, such as soot, chars, glassy carbon, and evaporated a-C, is shown in the lower left hand corner. If the fraction of sp3 bonding reaches a high degree, such an a-C is denoted as tetrahedral amorphous carbon (ta-C), in order to distinguish it from sp2 a-C [2]. Two hydrocarbon polymers, that is, polyethylene (CH2)n and polyacetylene (CH)n, define the limits of the triangle in the right hand corner beyond which interconnecting C-C networks do not form, and only strait-chain molecules are formed. The DLC films, i.e. a-C, ta-C, a-C:H and ta-C:H, have some extreme properties similar to diamond, such as hardness, elastic modulus and chemical inertness. These films are great advantages for many applications. One of the most important applications of the carbon-based films is the coating for magnetic hard disk recording. The second successful application is wear protective and antireflective films for IR windows. The third application is wear protection of bearings and sliding friction parts. The fourth is precision gages for the automotive industry. Recently, exciting ongoing study [1] tries to deposit a carbon-based protective film on engine parts (e.g. engine cylinders and pistons) taking into account not only low friction and wear, but also self lubricating properties. Reduction of the oil consumption is expected. Currently, for an additional application field, the carbon-based films are extensively studied as excellent candidates for biocompatible films on biomedical implants. The carbon-based films consist of carbon, hydrogen and nitrogen, which are biologically harmless as well as the main elements of human body. Some in vitro and limited in vivo studies on the biological effects of carbon-based films have been studied [$2{\sim}5$].The carbon-based films have great potentials in many fields. However, a few technological issues for carbon-based film are still needed to be studied to improve the applicability. Aisenberg and Chabot [3] firstly prepared an amorphous carbon film on substrates remained at room temperature using a beam of carbon ions produced using argon plasma. Spencer et al. [4] had subsequently developed this field. Many deposition techniques for DLC films have been developed to increase the fraction of sp3 bonding in the films. The a-C films have been prepared by a variety of deposition methods such as ion plating, DC or RF sputtering, RF or DC plasma enhanced chemical vapor deposition (PECVD), electron cyclotron resonance chemical vapor deposition (ECR-CVD), ion implantation, ablation, pulsed laser deposition and cathodic arc deposition, from a variety of carbon target or gaseous sources materials [5]. Sputtering is the most common deposition method for a-C film. Deposited films by these plasma methods, such as plasma enhanced chemical vapor deposition (PECVD) [6], are ranged into the interior of the triangle. Application fields of DLC films investigated from papers. Many papers purposed to apply for tribology due to the carbon-based films of low friction and wear resistance. Figure 1 shows the percentage of DLC research interest for application field. The biggest portion is tribology field. It is occupied 57%. Second, biomedical field hold 14%. Nowadays, biomedical field is took notice in many countries and significantly increased the research papers. DLC films actually applied to many industries in 2005 as shown figure 2. The most applied fields are mold and machinery industries. It took over 50%. The automobile industry is more and more increase application parts. In the near future, automobile industry is expected a big market for DLC coating. Figure 1 Research interests of carbon-based filmsFigure 2 Demand ratio of DLC coating for industry in 2005. In this presentation, I will introduce a trend of carbon-based coating research and applications.

• Journal of The Korean Society of Grassland and Forage Science
• /
• v.27 no.4
• /
• pp.249-256
• /
• 2007

This experiment was conducted to compare the agronomic characteristics and productivity in introduced hairy vetch cultivars and developed new Korean Hairy vetch cultivars in two areas of the experimental field of Grassland and Forage Crops Division, National Institute of Animal Science from 2005 to 2006. The experiment was arranged in randomized complete block design with three replications. A total of 11 hairy vetch cultivars used in this study including 9 introduced cultivars (Penn-02, Sander, Latigo, Welta, Ostsaat, VV4712, Minnie, Barlosa and Capello) and 2 Korean cultivars (Cold green and Cheong pa). The early flowering varieties of Minnie, Barlosa, Capello and Cold green were grown as early maturity cultivars, while the late flowering varieties of Penn-02, Sander, Latigo, Welta, Ostsaat, VV4712 and Cheong pa were grown as medium and late maturity. The winter hardiness of hairy vetch was good except for Minnie, Barlosa and Capello of early maturity cultivars. In this study, dry matter yield of VV4712 was the highest of 11 hairy vetch cultivars. And dry matter yield of medium and late maturity vetch increased in the middle region while that of early maturity increased in the south. The new Korean cultivar, Cheong pa, as a medium and late maturity hairy vetch was not bad compared to introduced cultivars and Cold green as an early maturity hairy vetch was an excellent cultivar compared to introduced cultivars in both regions. Latigo in NDF (neutral detergent fiber) and Welta in ADF (acid detergent fiber) were lower than other cultivars, while Sander in IVDMD (in vitro dry matter digestibility) and CP (crude protein) content were higher than other cultivars. The results of this experiment indicated that the productivity of hairy vetch cultivars was highest in VV4712, and Cheong pa and Cold green of new Korean cultivar were the promising cultivars of hairy vetch in Korea.

• Journal of Life Science
• /
• v.23 no.4
• /
• pp.501-509
• /
• 2013

Formaldehyde (FA) is widely used in industries, and it is an indoor and outdoor pollutant. Exposure to FA may cause inflammation and respiratory oxidative stress. Studies have demonstrated that FA can cause cancer in animal models. During the regeneration process of injured starfish (Asterina pectinifera), several changes have been observed in the expression of cytokines. In particular, higher TGF-${\beta}1$ expression has been detected in arm cut starfish extract after eight days. The current study was designed to elucidate the in-vitro and the in-vivo pharmacological effects of starfish extract on FA exposure. We investigated the protective effects of intact starfish extract and arm cut starfish extract on an IMR-90 cell line and on mouse lung injury in response to FA exposure. In the presence of FA, inhalation of the arm cut starfish extract was associated with more promising cell proliferation, TNF-${\alpha}$, NF-${\kappa}B$ decrement, and $I{\kappa}-B{\alpha}$ increment. In the experimental group, the pulmonary structure of the arm cut starfish extract-treated group in the presence of FA exposure was similar to the control group, whereas the FA exposure group showed damage to the pulmonary structure. Moreover, the arm cut starfish extracts was more effective than the intact starfish extracts in terms of the expression of TNF-${\alpha}$, NF-${\kappa}B$, $I{\kappa}-B{\alpha}$, and surfactant protein A. The results obtained in this study demonstrate that arm cut starfish extracts are more effective in protecting pulmonary structure and function against FA exposure than intact starfish extracts.

• Clinical and Experimental Reproductive Medicine
• /
• v.13 no.1
• /
• pp.11-19
• /
• 1986

Follicular flxid (FF) and their matched oocytes were obtained from 58 follicles of 27 women who underwent an in vitro fertilization (IVF) procedure with ovarian hyperstimulation by clomiphene citrate(n=8), hMG(n=9),FSH/hMG(n=10). Follicular aspiration was performed 36 hours after human chorionic gonadotropin administration. The concentcation of androstendione (ADD), testosterone (T) was correlated with hyperstimulation regimens, the morphology of the oocyte-corona-cumulus complex (OCCC), oocyte fertilization, and the incidence of pregnancy after embryo transfer. The results were as follows. 1. According to hyperstimulation regimens, there was no significant differance in FF ADD and T concentrations of the similar morphology of OCCC. 2. In clomiphene-treated and FSH/hMG-treated cycles, FF ADD concentrations of preovulatory oocytes were 43.09${\pm}$9.53 ng/ml and 59.46${\pm}$9.09 ng/ml, those of immature occytes were 96.98${\pm}$16.55 ng/ml and 116.13${\pm}$36.81 ng/ml, those of atretic oocytes were 246.5 ${\pm}$9.25 ng/ml and 634.25${\pm}$9.25 ng/ml respectively, reflecting the significant relationship between FF ADD level and morphologic maturity of OCCC (p<0.05). But in hMG-treated cycles, such relationship was not found (p>0.1). In clomiphene-treated and FSH/hMG-treated cycles, FF T concentrations of preovulatory oocytes were 11.37${\pm}$2.38 ng/ml and 11.68${\pm}$1.73 ng/ml respectively which were significantly lower than those of atretic oocytes (25.1${\pm}$7.50 ng/ml and 23.25${\pm}$0.95 ng/ml respectively) (p<0.05). But in all cycles, FF T concentrations of immature oocytes were not significantly different from those of preovulatory oocytes, artetic oocytes (p>0.1). 3. In hMG-treated and FSH/hMG-treated cycles, FF ADD concentrations of fertilized oocytes were 32.43${\pm}$4.09 ng/ml and 42.61${\pm}$4.82 ng/ml respectively which were significantly lower than those of non-fertilized oocytes (72.18${\pm}$17.31 ng/ml and 108.09${\pm}$17.32 ng/ml respectively) (p<0.05), but in clomiphene-treated cycles there was no significant difference (p>0.1). In FSH/hMG-treated cycles, FF T concentration of fertilized oocytes was 7.33${\pm}$1.06 ng/ml which was significantly lower than that of non-fertilized oocytes (20.3${\pm}$6.21 ng/ml) (p>0.02), but in clomiphne-treated and hMG-treated cycles there was no significant difference (p>0.1). 4. In all cycles FF ADD and T concentrations did not correlated with the success of pregnancy after embryo transfer. Above results suggested that FF ADD and T may play an important role in oocyte maturation and fertilization, but their relationship with the success of psegnancy was not found.

• Journal of Life Science
• /
• v.16 no.4
• /
• pp.571-578
• /
• 2006

HtrA2/Omi, a mitochondrial trypsin-like serine protease, is pivotal in regulating apoptotic cell death. Several lines of recent evidence suggest that HtrA2 is associated with the pathogenesis of neurodegenerative disorders; however, the physiological function of HtrA2 still remains elusive. For studying physiological function of HtrA2 in depth, it is necessary to develop a suitable expression system in the model animal. We therefore utilized the zebrafish as a model animal to establish expression of human HtrA2 (hHtrA2) in vivo. For expression of mature HtrA2 as GFP fusion in zebrafish embryos, the HtrA2 (WT) or (S306A) cDNAs with the C-terminal GFP tag were inserted into the pCS2+ plasmid. Expression patterns of HtrA2 in HEK293 cells were first monitored by immunofluorescence staining and immunoblot assays, showing approximately 64 kDa of the HtrA2-GFP fusion proteins. Subsequently, the hHtrA2 plasmid DNA or in vitro transcribed mRNA was microinjected into zebrafish embryos. The expression patterns of HtrA2 in Zebrafish embryos were monitored by GFP fluorescence in 24 hours-post-fertilization (hpf). Although expression patterns of HtrA2-GFP in developing embryos were different between the injected DNA and mRNA, both nucleic acids revealed good expression levels to further study the physiological role of HtrA2 in vivo. This study provides a suitable condition for expressing hHtrA2 in the zebrafish embryos as well as a method for generating useful system to investigate physiological properties of the specific human genes.

• Proceedings of the Korean Society of Applied Pharmacology
• /
• 2007.11a
• /
• pp.79-92
• /
• 2007

Oxidative stress have known to be a risk factor for the degenerative processes and closely related to a lot of diseases. It is well established that antioxidants are good in protection and therapeutic means against oxidative damage. There is increasing interest in natural antioxidants and many natural antioxidants have been found and utilized as the possible protection for various diseases and skin aging. We have screened natural antioxidant agents for cosmeceuticals, nutraceuticals, and drugs as therapeutic and preventive means against oxidative stress, and have developed a number of novel antioxidants from various natural sources. A novel melanin synthesis inhibitor, Melanocin A, isolated from the metabolite of a fungal strain Eupenicillium shearii F80695 inhibited mushroom tyrosinase and melanin biosynthesis of B16 melanoma cells with $IC_{50}$ value of 9.0 nM and MIC value of $0.9\;{\mu}M$, respectively. Melanocin A also exhibited potent antioxidant activity by scavenging of DPPH and superoxide anion radicals. UV was found to increase the level of hydrogen peroxides and other reactive oxygen species (ROS) in skin tissues. This increase in ROS may not only alter the structure and function of many genes and proteins directly but may also modulate their expressions through signal transduction pathways and, ultimately, lead to skin damage. We investigated the effect of Melanocin A on UV-induced premature skin aging. Firstly, the effect of Melanocin A on UV-induced matrix metalloproteinase (MMP)-9 expression in an immortalized human keratinocyte cell line, HaCaT in vitro was investigated. Acute UV irradiation induced MMP-9 expression at both the mRNA and protein levels and Melanocin A suppressed this expression in a dose-dependent manner. We then investigated UV-induced skin changes in hairless mice in vivo by Melanocin A. Chronic exposure of hairless mouse dorsal skin to UV increased skin thickness and induced wrinkle formation and the gelatinase activities of MMP-2 and MMP-9. Moreover, Melanocin A significantly suppressed UV-induced morphologic skin changes and MMP-2 and MMP-9 expression. These results show that Melanocin A can prevent the harmful effects of UV that lead to skin aging. Therefore, we suggest that Melanocin A should be viewed as a potential therapeutic agent for preventing and/or treating premature skin aging. Terrein is a bioactive fungal metabolite isolated from Penicillium species. Terrein has a relatively simple structure and can be easily synthesized. However, the biologic effects of terrein are comparatively unknown. We found for the first time that terrein potently inhibit melanin production in melanocytes and has a strong hypopigmentary effect in a spontaneously immortalized mouse melanocyte cell line, Mel-Ab. Treatment of Mel-Ab cells with terrein (10-100 mM) for 4 days significantly reduced melanin levels in a dose-dependent manner. In addition, terrein at the same concentration also reduced tyrosinase activity. We then investigated whether terrein influences the extracellular signal-regulated protein kinase (ERK) pathway and the expression of microphthalmia-associated transcription factor (MITF), which is required for tyrosinase expression. Terrein was found to induce sustained ERK activation and MITF down-regulation, and luciferase assays showed that terrein inhibits MITF promoter activity in a dose-dependent manner. To elucidate the correlation between ERK pathway activation and a decreased MITF transcriptional level, PD98059, a specific inhibitor of the ERK pathway, was applied before terrain treatment and found to abrogate the terrein-induced MITF attenuation. Terrein also reduced the tyrosinase protein level for at least 72 h. These results suggest that terrain reduces melanin synthesis by reducing tyrosinase production via ERK activation, and that this is followed by MITF down-regulation.

• Journal of Preventive Medicine and Public Health
• /
• v.28 no.4 s.51
• /
• pp.863-873
• /
• 1995

This report describes a preperation and characterization of canine blood lead(Pb) standard reference material(SRM). Three adult beagle dogs(A, B, and C)were orally dosed with gelatin capsules containing $Pb(NO_3)_2$, equivalent to $10\sim80mg$ Pb/kg body weight. Blood was drawn 24 hours after the dose from the cephalic vein into lead free 500ml Pyrex beaker in which EDTA.K was contained as an anticoagulant. The amount of lead given to individual dog was varied arbitrarily. Three month later, 3 canine animals were orally dosed with lead secondarily to make mixed SRM(D1) which was mixed different concentrations of lead in bloods with A1, B1, and C1 in vitro. The SRMs for A, B, C, A1, B1, C1, and D1 were distributed 2ml each into more than 300 lead free bottles, and were stored in refregerator at $4^{\circ}C$. The amount of lead in canine whole blood samples were determined using a Varian 30A atomic absorption spectrophotometer(AAS) with a model GTA-96 graphite tube atomizer with D2 background correction and a Hitachi Z-8100 AAS with Zeeman background correction. The sensitivity and detection limits for lead determination of Varian 30A were $0.46{\mu}g/L,\;0.34{\mu}g/L,\;and\;0.56{\mu}g/L,\;0.14{\mu}g/L$ of Hitachi Z-8100, respectively. Day to day variations in determination of blood lead concentration in a certain sample were $31.11{\pm}1.36{\mu}g/100ml$ by Varian 30A, and $33.08{\pm}0.82{\mu}g/100ml$ by Hitachi Z-8100, showing the difference of 3% between the two results. At the blood lead concentrations of $56.31{\pm}1.98{\mu}g/100ml(A),\;40.89{\pm}0.80{\mu}g/100ml(B),\;59.01{\pm}1.38{\mu}g/100ml(C)$, the precisions of replicated measurements by AAS were 3.52%, 1.96%, and 2.34%, respectively. Coefficient variation(CV) of SRMs(A, B, and C) within a standard sample were ranged from 0.92% to 7.50%, and those between 5 standard samples were 1.21%, 2.64%, and 1.11%, respectively, showing inter-vial variation of $1{\mu}g/100ml$. Lead levels in SRMs during one month storage were unchanged. The overall recoveries were $89.6\sim100.4%,\;91.6\sim101.9%,\;90.3\sim100.0%$ for A, B, and C SRMs, means were $56.46{\pm}2.69{\mu}g/100ml,\;39.35{\pm}1.89{\mu}g/100ml,\;57.40{\pm}2.31{\mu}g/100ml$, and measurement ranges were$52.88{\pm}59.26{\mu}g/100ml,\;37.47{\pm}41.68{\mu}g/100ml,\;54.80{\pm}60.69{\mu}g/100ml$, respectively. Those results were laid within confidence limits values. The lead concentrations in the mixed sample(D1) stored over one month period were ranged from $32.76{\mu}g/100ml\;to\;33.54{\mu}g/100ml$, with CV ranging from 1.2% to 2.7%. The results were similiar to each of single samples(A1, B1, and C1) in respect of homogeneity and stability. Results of the mixed blood sample analysed after 1 month storage at $4^{\circ}C$ by four other laboratories(L1, L2, L3, L4) were similar with those of our laboratory($L5;31.18{\pm}0.24{\mu}g/100ml$, acceptable range by $CDC;25.18\sim37.18{\mu}g/100ml$), showing the concentrations of $25.91{\pm}1.19{\mu}g/100ml(L1),\;34.16{\pm}0.22{\mu}g/100ml(L2),\;35.68{\pm}0.85{\mu}g/100ml(L3),\;30.95{\pm}0.46{\mu}g/100ml(L4)$ in a each samples.

• Journal of Life Science
• /
• v.23 no.2
• /
• pp.167-174
• /
• 2013

Peroxiredoxin 6 (Prx 6) is a member of the thiol-specific antioxidant protein family, which may play a role in protection against oxidative stress and in regulating phospholipid turnover. The aim of this study was to determine whether a human Prx 6/Luc vector was stably expressed and responded to antioxidants in a lung cell line (NCI-H460). To achieve this, the luciferase signal, hPrx 6 mRNA expression, and superoxide dismutase (SOD) activity were measured in transfectants with a hPrx 6/Luc plasmid after treatment with four antioxidant extracts, including Korea white ginseng (KWG), Korea red ginseng (KRG), Liriope platyphylla (LP), and red Liriope platyphylla (RLP). First, the hPrx 6/Luc plasmid was successfully constructed with DNA fragments of human Prx 6 promoter, amplified by PCR using genomic DNA isolated from NCI-H460 cells, and cloned into the pTransLucent reporter vector. The orientation and sequencing of the hPrx 6/Luc plasmid were identified with restriction enzyme and automatic sequencing. A luciferase assay revealed significant enhancement of luciferase activity in the four treatment groups compared with a vehicle-treated group, although the ratio of the increase was different within each group. The KRG- and LP-treated groups showed higher activity than the KWG- and RLP-treated groups. Furthermore, the luciferase activity against RLP occurred roughly in a dose-dependent manner. However, the level of endogenous hPrx 6 mRNA did not change in any group treated with the four extracts. The SOD activity was in agreement with the luciferase activity. Therefore, these results indicate that the hPrx 6/Luc vector system may successfully express and respond to antioxidant compounds in NCI-H460 cells. The data also suggest that the Prx 6/Luc vector system may be effectively applied in screening the response of hPrx 6 to antioxidant compounds in transgenic mice.

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.44 no.4
• /
• pp.506-515
• /
• 2015

In this study, the effect of Goheung pomegranate extract on postmenopausal syndrome was evaluated both in vitro and in vivo in ovariectomized Sprague-Dawly (SD) rats. Sixty female SD rats were divided into six groups: sham, sham operation and distilled water; OVX, ovariectomized and distilled water; PE1, ovariectomized and pomegranate concentrate (0.75 mL/twice/d); PE2, ovariectomized and pomegranate concentrate (1.5 mL/twice/d); PE3, ovariectomized and pomegranate concentrate (2.2 mL/twice/d); and CE, ovariectomized and commercial pomegranate concentrate (2.2 mL/twice/d). Percent bone volume (bone volume/tissue volume) and trabecular thickness (Tb.Th) improved in a dose-independent manner in PE1, 2, and 3. Especially, bone mineral density was significantly improved in PE3 (P<0.05) compared to OVX. Pomegranate extract reduced body weight and visceral fat mass. High density lipoprotein cholesterol (HDL-C) level slightly increased in a dose-independent manner in the experimental group. In addition, HDL-C/total cholesterol level of PE3 significantly increased (P<0.05) compared with OVX. These results show that pomegranate concentrate improved blood lipid levels and bone metabolism in ovariectomized rats. Therefore, Goheung pomegranate concentrates are expected to improve cardiovascular and bone-related diseases in menopausal women.