• Title/Summary/Keyword: Reaction-time

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Processed Panax ginseng, sun ginseng, inhibits the differentiation and proliferation of 3T3-L1 preadipocytes and fat accumulation in Caenorhabditis elegans

  • Lee, Hyejin;Kim, Jinhee;Park, Jun Yeon;Kang, Ki Sung;Park, Joeng Hill;Hwang, Gwi Seo
    • Journal of Ginseng Research
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    • v.41 no.3
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    • pp.257-267
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    • 2017
  • Background: Heat-processed ginseng, sun ginseng (SG), has been reported to have improved therapeutic properties compared with raw forms, such as increased antidiabetic, anti-inflammatory, and antihyperglycemic effects. The aim of this study was to investigate the antiobesity effects of SG through the suppression of cell differentiation and proliferation of mouse 3T3-L1 preadipocyte cells and the lipid accumulation in Caenorhabditis elegans. Methods: To investigate the effect of SG on adipocyte differentiation, levels of stained intracellular lipid droplets were quantified by measuring the oil red O signal in the lipid extracts of cells on differentiation Day 7. To study the effect of SG on fat accumulation in C. elegans, L4 stage worms were cultured on an Escherichia coli OP50 diet supplemented with $10{\mu}g/mL$ of SG, followed by Nile red staining. To determine the effect of SG on gene expression of lipid and glucose metabolism-regulation molecules, messenger RNA (mRNA) levels of genes were analyzed by real-time reverse transcription-polymerase chain reaction analysis. In addition, the phosphorylation of Akt was examined by Western blotting. Results: SG suppressed the differentiation of 3T3-L1 cells stimulated by a mixture of 3-isobutyl-1-methylxanthine, dexamethasone, and insulin (MDI), and inhibited the proliferation of adipocytes during differentiation. Treatment of C. elegans with SG showed reductions in lipid accumulation by Nile red staining, thus directly demonstrating an antiobesity effect for SG. Furthermore, SG treatment down-regulated mRNA and protein expression levels of peroxisome proliferator-activated receptor subtype ${\gamma}$ ($PPAR{\gamma}$) and CCAAT/enhancer-binding protein-alpha ($C/EBP{\alpha}$) and decreased the mRNA level of sterol regulatory element-binding protein 1c in MDI-treated adipocytes in a dose-dependent manner. In differentiated 3T3-L1 cells, mRNA expression levels of lipid metabolism-regulating factors, such as amplifying mouse fatty acid-binding protein 2, leptin, lipoprotein lipase, fatty acid transporter protein 1, fatty acid synthase, and 3-hydroxy-3-methylglutaryl coenzyme A reductase, were increased, whereas that of the lipolytic enzyme carnitine palmitoyltransferase-1 was decreased. Our data demonstrate that SG inversely regulated the expression of these genes in differentiated adipocytes. SG induced increases in the mRNA expression of glycolytic enzymes such as glucokinase and pyruvate kinase, and a decrease in the mRNA level of the glycogenic enzyme phosphoenol pyruvate carboxylase. In addition, mRNA levels of the glucose transporters GLUT1, GLUT4, and insulin receptor substrate-1 were elevated by MDI stimulation, whereas SG dose-dependently inhibited the expression of these genes in differentiated adipocytes. SG also inhibited the phosphorylation of Akt (Ser473) at an early phase of MDI stimulation. Intracellular nitric oxide (NO) production and endothelial nitric oxide synthase mRNA levels were markedly decreased by MDI stimulation and recovered by SG treatment of adipocytes. Conclusion: Our results suggest that SG effectively inhibits adipocyte proliferation and differentiation through the downregulation of $PPAR{\gamma}$ and $C/EBP{\alpha}$, by suppressing Akt (Ser473) phosphorylation and enhancing NO production. These results provide strong evidence to support the development of SG for antiobesity treatment.

Purification of Human HtrA1 Expressed in E. coli and Characterization of Its Serine Protease Activity (E. coli에서 발현된 human HtrA1 단백질의 정제와 HtrA1의 serine protease 활성 조건에 관한 연구)

  • Kim, Kyung-Hee;Kim, Sang-Soo;Kim, Goo-Young;Rhim, Hyang-Shuk
    • Journal of Life Science
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    • v.16 no.7 s.80
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    • pp.1133-1140
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    • 2006
  • Human HtrA1 (High temperature requirement protein A1) is a homologue of the E. coli periplasmic serine protease HtrA. A recent study has demonstrated that HtrA1 is a serine protease involved in processing of insulin like growth factor binding protein (ICFBP), indicating that it serves as an important regulator of IGF activity. Additionally, several lines of evidence suggest a striking correlation between proteolytic activity of HtrA1 serine protease and the pathogenesis of several diseases; however, physiological roles of HtrA1 remain to be elucidated. We used the pGEX bacterial expression system to develop a simple and rapid method for purifying HtrA1, and the recombinant HtrA1 protein was utilized to investigate the optimal conditions in executing its proteolytic activity. The proteolytically active HtrA1 was purified to approximately 85% purity, although the yield of the recombinant HtrA1 protein was slightly low $460{\mu}g$ for 1 liter E. coli culture). Using in vitro endoproteolytic cleavage assay, we identified that the HtrA1 serine protease activity was dependent on the enzyme concentration and the incubation time and that the best reaction temperature was $42^{\circ}C$ instead of $37^{\circ}C$. We arbitrary defined one unit of proteolytic activity of the HtrA1 serine protease as 200nM of HtrA1 that cleaves half of $5{\mu}M\;of\;{\beta}-casein$ during 3 hr incubation at $37^{\circ}C$. Our study provides a method for generating useful reagents to investigate the molecular mechanisms by which HtrA1 serine protease activity contributes in regulating its physiological function and to identify natural substrates of HtrA1.

Reactive Oxygen Species Mediates Lysophosphatidic Acid-induced Migration of SKOV-3 Ovarian Cancer Cells (SKOV-3 난소암 세포주에서 lysophosphatidic acid 유도 세포의 이동에 있어 활성산소의 역할)

  • Kim, Eun Kyoung;Lee, Hye Sun;Ha, Hong Koo;Yun, Sung Ji;Ha, Jung Min;Kim, Young Whan;Jin, In Hye;Shin, Hwa Kyoung;Bae, Sun Sik
    • Journal of Life Science
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    • v.22 no.12
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    • pp.1621-1627
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    • 2012
  • Cell motility plays an essential role in many physiological responses, such as development, immune reaction, and angiogenesis. In the present study, we showed that lysophosphatidic acid (LPA) modulates cancer cell migration by regulation of generation of reactive oxygen species (ROS). Stimulation of SKOV-3 ovarian cancer cells with LPA strongly promoted migration. but this migration was completely blocked by pharmacological inhibition of phosphatidylinositol 3-kinase (PI3K)/Akt signaling pathway. Inhibition of the ERK pathway had no effect on migration. Stimulation of SKOV-3 ovarian cancer cells with LPA significantly induced the generation of ROS in a time-dependent manner. LPA-induced generation of ROS was significantly blocked by pharmacological inhibition of PI3K or Akt, but inhibition of the ERK signaling pathway had little effect. LPA-induced generation of ROS was blocked by pretreatment of SKOV-3 ovarian cancer cells with an NADPH oxidase inhibitor, whereas inhibition of xanthine oxidase, cyclooxygenase, or mitochondrial respiratory chain complex I had no effect. Scavenging of ROS by N-acetylcysteine completely blocked LPA-induced migration of SKOV-3 ovarian cancer cells. Inhibition of NADPH oxidase blocked LPA-induced migration whereas inhibition of xanthine oxidase, cyclooxygenase, or mitochondrial respiratory chain complex I did not affect LPA-induced migration of SKOV-3 ovarian cancer cells. Given these results, we suggest that LPA induces ROS generation through the PI3K/Akt/NADPH oxidase signaling axis, thereby regulating cancer cell migration.

Characteristics of Diurnal Variation of High PM2.5 Concentration by Spatio-Temporal Wind System in Busan, Korea (시·공간적 풍계에 따른 부산지역 고농도 PM2.5의 일변화 특성)

  • Kim, Bu-Kyung;Lee, Dong-In;Kim, Jeong-Chang;Lee, Jun-Ho
    • Journal of the Korean earth science society
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    • v.33 no.6
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    • pp.469-480
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    • 2012
  • This study was to analyze the characteristics of diurnal variation of high $PM_{2.5}$ concentration, $PM_{2.5}/PM_{10}$ concentration ratio by spatio-temporal wind system (wind speed and wind direction) for high $PM_{2.5}$ concentration (over the 24 hr environmental standard of $PM_{2.5}$, $50{\mu}g/m^3$) in the air quality observation sites (Jangrimdong: Industrial area, Jwadong: Residential area) that were measured for 3 years (2005. 12. 1-2008. 11. 30) in Busan. The observation days of high $PM_{2.5}$ concentration were 182 at Jangrimdong and 27 at Jwadong. The seasonal diurnal variation of hourly mean of high $PM_{2.5}$ concentration and of $PM_{2.5}/PM_{10}$ concentration ratio showed a similar pattern that had higher variation at dawn, and night and in the morning than in the afternoon. Durning daytime in summer at Jwadong, the $PM_{2.5}/PM_{10}$ concentration ratio increased because a secondary particulate matter, which was created by photochemical reaction, decreased the coarse particles of $PM_{10}$ more than the fine particles of $PM_{2.5}$ concentrations in ocean condition. We did an analysis of spatio-temporal wind system (wind speed range and wind direction) in each time zone. The result showed that high $PM_{2.5}$ concentration at Jangrimdong occurred due to the congestion of pollutants emissions from the industrial complex in Jangrimdong area and the transportation of pollutants from places nearby Jangrimdong. It also showed that high $PM_{2.5}$ concentration occurred at Jwadong because of a number of local residential and commercial activities that caused the congestion of pollutants.

Study on Synthesis of Pyrochlore in Gd-Ti-O and Gd-Zr-O Systems (Gd-Ti-O계 및 Gd-Zr-O 계에서의 파이로클로어 합성연구)

  • ;;;S.V. Yudintsev
    • Economic and Environmental Geology
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    • v.37 no.3
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    • pp.303-309
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    • 2004
  • Pyrochlores were known as promising materials for the immobilization of radioactive actinide. Accordingly, we synthesized pyrochlores with Gd$_2$Ti$_2$$O_7$ and Gd$_2$Zr$_2$$O_7$compositions by sintering method, and studied its properties and phase relations in Gd-Ti-O and Gd-Zr-O system. The mixed powders were pressed into pellets under 200-400 kgf/cm$^2$ at room temperature. and then sintered at 1000-1$600^{\circ}C$ for 0.5-40 hours. The synthesized samples were analyzed and were identified with XRD and SEM/EDS analyses. The optimal synthetic conditions of pyrochlores with Gd$_2$Ti$_2$$O_7$composition were at 140$0^{\circ}C$/0.5hrs, 130$0^{\circ}C$/3hrs and 120$0^{\circ}C$/20hrs. Its chemical composition was $Gd_{2.0-2.1}$$Ti_{1.9-2.0}$$O_7$ and similar to the stoichiometric composition without any relationship in temperature and atmosphere. The optimal synthetic conditions of pyrochlores with $Gd_{2}$$Zr_{2}$$O_7$composition were at 155$0^{\circ}C$/40hrs and 1$600^{\circ}C$/30hrs. The compositions of pyrochlore synthesized from these optimal conditions were irregular with $Gd_{1.5-2.4}$$Zr_{1.7-2.4}$$O_7$. Such heterogeneity indicates that the reaction rate of pyrochlore with Gd$_2$Zr$_2$$O_7$composition is very low, and then its equilibrium state could not be attained even for 40 hours which was the longest sintering time in this research.

The Individual Discrimination Location Tracking Technology for Multimodal Interaction at the Exhibition (전시 공간에서 다중 인터랙션을 위한 개인식별 위치 측위 기술 연구)

  • Jung, Hyun-Chul;Kim, Nam-Jin;Choi, Lee-Kwon
    • Journal of Intelligence and Information Systems
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    • v.18 no.2
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    • pp.19-28
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    • 2012
  • After the internet era, we are moving to the ubiquitous society. Nowadays the people are interested in the multimodal interaction technology, which enables audience to naturally interact with the computing environment at the exhibitions such as gallery, museum, and park. Also, there are other attempts to provide additional service based on the location information of the audience, or to improve and deploy interaction between subjects and audience by analyzing the using pattern of the people. In order to provide multimodal interaction service to the audience at the exhibition, it is important to distinguish the individuals and trace their location and route. For the location tracking on the outside, GPS is widely used nowadays. GPS is able to get the real time location of the subjects moving fast, so this is one of the important technologies in the field requiring location tracking service. However, as GPS uses the location tracking method using satellites, the service cannot be used on the inside, because it cannot catch the satellite signal. For this reason, the studies about inside location tracking are going on using very short range communication service such as ZigBee, UWB, RFID, as well as using mobile communication network and wireless lan service. However these technologies have shortcomings in that the audience needs to use additional sensor device and it becomes difficult and expensive as the density of the target area gets higher. In addition, the usual exhibition environment has many obstacles for the network, which makes the performance of the system to fall. Above all these things, the biggest problem is that the interaction method using the devices based on the old technologies cannot provide natural service to the users. Plus the system uses sensor recognition method, so multiple users should equip the devices. Therefore, there is the limitation in the number of the users that can use the system simultaneously. In order to make up for these shortcomings, in this study we suggest a technology that gets the exact location information of the users through the location mapping technology using Wi-Fi and 3d camera of the smartphones. We applied the signal amplitude of access point using wireless lan, to develop inside location tracking system with lower price. AP is cheaper than other devices used in other tracking techniques, and by installing the software to the user's mobile device it can be directly used as the tracking system device. We used the Microsoft Kinect sensor for the 3D Camera. Kinect is equippedwith the function discriminating the depth and human information inside the shooting area. Therefore it is appropriate to extract user's body, vector, and acceleration information with low price. We confirm the location of the audience using the cell ID obtained from the Wi-Fi signal. By using smartphones as the basic device for the location service, we solve the problems of additional tagging device and provide environment that multiple users can get the interaction service simultaneously. 3d cameras located at each cell areas get the exact location and status information of the users. The 3d cameras are connected to the Camera Client, calculate the mapping information aligned to each cells, get the exact information of the users, and get the status and pattern information of the audience. The location mapping technique of Camera Client decreases the error rate that occurs on the inside location service, increases accuracy of individual discrimination in the area through the individual discrimination based on body information, and establishes the foundation of the multimodal interaction technology at the exhibition. Calculated data and information enables the users to get the appropriate interaction service through the main server.

An Outbreak of Epidemic Keratoconjunctivitis by Adenovirus Type 8 in a Neonatal Intensive Care Unit (신생아 중환자실에서의 아데노바이러스 8형에 의한 유행성 각결막염의 발생)

  • Park, Na-Ri-Mi;Na, Ji-Youn;Joung, Kyoung-Eun;Lee, Ji-Na;Kim, Ee-Kyung;Kim, Han-Suk;Kim, Seong-Joon;Song, Jung-Suk;Oh, Hyang-Soon;Lee, Hoan-Jong;Choi, Jung-Hwan
    • Neonatal Medicine
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    • v.15 no.1
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    • pp.44-53
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    • 2008
  • Purpose : Epidemic keratoconjunctivitis (EKC) caused by adenovirus is a highly contagious disease, which has been reported as outbreaks involving adults in the community. However, there has been no report on EKC outbreak by adenovirus in a neonatal intensive care unit (NICU) in Korea. Aims of this study were to investigate the EKC outbreak by adenovirus type 8 in NICU and to confirm an effectiveness of polymerase chain reaction (PCR) for diagnosis. Methods : Conjunctival swab or nasopharyngeal aspirate specimens were taken from all patients and tested by viral culture and PCR. Adenovirus serotype was determined by sequencing of PCR product of selected region of hexon gene using the virus isolates or specimens. Results : An outbreak of EKC occurred which was involving 12 preterm infants in the NICU of the Seoul National University Children's Hospital between July 12th and August 1st, 2005. Three hospital staffs and one family member of the neonate were also affected. Adenovirus was detected in 12/12 (100%), 6/11 (54.5%) by PCR and virus culture, respectively. Eleven PCR-positive neonates were identified as serotype 8 by sequencing. The first affected 4 babies have had routine ROP (retinopathy of prematurity) examinations one week ago. While previous outbreaks were sustained for a few months, the event in our unit was controlled without complications in 3 weeks. Conclusion : We analyzed the EKC outbreak by adenovirus type 8 in NICU. Adenovirus serotype was identified by PCR and sequencing with high sensitivity for the first time in Korea, so we suggest this method can be very useful for rapid diagnosis and infection control.

Understanding the Response Characteristics of X-ray Verification Film (X-선 Verification 필름의 반응 특성에 관한 연구)

  • Yeo Inhwan;Seong Jinsil;Chu Sung Sil;Kim Gwi Eon;Suh Chang Ok;Burch Sandra E.;Wang Chris K.
    • Radiation Oncology Journal
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    • v.16 no.4
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    • pp.505-515
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    • 1998
  • Purpose : This study is intended to understand the sensitometric characteristics and the emulsion properties of the commercially available CEA TVS film in comparison with the Kodak X-Omat V film. Materials and Methods : For this purpose, we have formulated an analytic expression of the characteristic curves for x-ray film exposed to mixed radiation of electrons, photons, and visible light. This mathematical expression was developed based on reaction-rate and target-hit theories. Unlike previous expressions. it relates optical density to emulsion properties such as grain size and silver bromide content We have also developed a quantity which characterizes the film response to visible light relative to that to photons and electrons. This quantity could be expressed as a function of grain area. Thus, we have developed mathematical expressions and quantities with which the emulsion properties of the films can be revealed based on the sensitometric characteristics. Demonstrating the use of this analytical study, we exposed CEA and Kodak verification films to the mixed radiation of electrons, photons, and visible light, and interpreted the experimental results accordingly. Results : We have demonstrated that: (1) the saturation density increases as the silver bromide content increases, (2) the time required to reach the threshold dose (to which the film begins to respond) when films are exposed to visible light decreases as the grain size increases, and (3) the CEA film contains more silver bromide. whereas the Kodak film contains larger grains. These findings were supported by the data provided by the manufacturers afterward. Conclusion : This study presented an analytical and experimental basis for understanding the response of X-ray film with respect to the emulsion properties.

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Expression of Yippee-Like 5 (YPEL5) Gene During Activation of Human Peripheral T Lymphocytes by Immobilized Anti-CD3 (인체 말초혈액의 활성화 과정 중 yippee-like 5 (YPEL5) 유전자의 발현 양상)

  • Jun, Do-Youn;Park, Hye-Won;Kim, Young-Ho
    • Journal of Life Science
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    • v.17 no.12
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    • pp.1641-1648
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    • 2007
  • Yippee-like proteins, which have been identified as the homolog of Drosophila yippee protein containing a zinc-finger domain, are known to be highly conserved among eukaryotes. However, their functional roles are still poorly understood. Recently we initiated ordered differential display (ODD)-polymerase chain reaction (PCR) to isolate genes of which expressions are altered following activation of human T cells. On the ODD-PCR image, one PCR-product detected in unstimulated T cells was not detectable at the time when the activated T cells traversed near $G_1/S$ boundary following activation by immobilized anti-CD3. Cloning and nucleotide sequence analysis revealed that the PCR-product was yippee-like 5 (YPEL5) gene, which was known as a human homolog of the Drosophila yippee gene. Northern blot analysis confirmed the amount of ${\sim}2.2$ kb YPEL5 mRNA expression detectable in unstimulated T cells was sustained until 1.5 hr after activation and then rapidly declined to undetectable level by 5 hr. Ectopic expression of YPEL5 gene in human cervix epitheloid carcinoma HeLa cells caused a significant reduction in cell proliferation to the level of 47% of the control. Expression of GFP-YPEL5 fusion protein in HeLa cells showed its nuclear localization. These results demonstrated that the expression level of human YPEL5 mRNA was negatively regulated in the early stage of T cell activation, and suggested that YPEL5 might exert an inhibitory effect on the cell proliferation as a nuclear protein.

Effects of Salt and Soysauce Condiment on Lipid Oxidation in Broiled Mackerel (Scomber japonicus) (소금과 간장 양념이 고등어 구이의 지질산화 안정성에 미치는 영향)

  • Ryu, Seung-Hee;Lee, Young-Soon;Moon, Gap-Soon
    • Korean Journal of Food Science and Technology
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    • v.34 no.6
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    • pp.1030-1035
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    • 2002
  • Despite health benefits derived from fish oil, polyunsaturated fatty acids (PUFAs) contained in fish oil are susceptible to lipid oxidation. To determine the optimum condition for maintaining good quality cooked fish during storage, mackerels were broiled with salt or soysauce condiments, and the lipid oxidation during 12 days of storage at refrigerated condition was measured. Peroxide value of broiled mackerel group with salt significantly increased after immediate cooking and maintained higher value throughout the storage period compare to the soysauce-added group, but showed similar value to the control group. Conjugated diene content in the soysauce-group was lower than the control and salt-added groups. Malondialdehyde content of broiled mackerel increased twofold and showed similar values in soysauce-added and the control groups during storage, whereas increased in the salt-added group significantly. Fatty acid compositions of the three mackerel groups changed after cooking, whereas that of the control group was almost stable during storage. In comparison with raw mackerel, the ratio of PUFA and saturated fatty acids decreased significantly, and the content of n-3 family fatty acid decreased from 25.53 to 20.63% in salted broiled mackerel. Soysauce group showed no reduction of PUFA with increasing storage time and showed the highest ratio of n-3/n-6 among the three groups at 10 days storage. Results reveal soysauce condiment protects against lipid peroxidation of broiled mackerel. Maillard reaction products (MRPs) found in soysauce might be responsible for the inhibitory effect and is a good condiment for extending storage life of cooked fish containing high amount of PUFA.