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Gene Expression Profiles in Cervical Cancer with Radiation Therapy Alone and Chemo-radiation Therapy (자궁경부암의 방사선치료 및 방사선항암화학 병용치료에 따른 유전자발현 조절양상)

  • Lee Kyu Chan;Kim Meyoung-kon;Kim Jooyoung;Hwang You Jin;Choi Myung Sun;Kim Chul Yong
    • Radiation Oncology Journal
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    • v.21 no.1
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    • pp.54-65
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    • 2003
  • Purpose : To analyze the gene expression Profiles of uterine ceulcal cancer, and its variation after radiation therapy, with or without concurrent chemotherapy, using a CDNA microarray. Materials and Methods :Sixteen patients, 8 with squamous ceil carcinomas of the uterine cervix, who were treated with radiation alone, and the other 8 treated w14h concurrent chemo-radiation, were Included in the study. Before the starling of the treatment, tumor biopsies were carried out, and the second time biopsies were peformed after a radiation dose of 16.2$\~$27 Gy. Three normal cervix tissues were used as a control group. The microarray experiments were peformed with 5 groups of the total RNAs extracted individually and then admixed as control, pre-radiation therapy alone, during-radiation therapy alone, pre-chemoradiation therapy, and during-chemoradlation therapy. The 33P-iabeled CDNAS were synthesized from the total RNAs of each group, by reverse transcription, and then they were hybridized to the CDNA microarray membrane. The gene expression of each microarrays was captured by the intensity of each spot produced by the radioactive isotopes. The pixels per spot were counted with an Arrayguage, and were exported to Microsoft Excel The data were normalized by the Z transformation, and the comparisons were peformed on the Z-ratio values calculated. Results : The expressions of 15 genes, including integrin linked kinase (ILK), CDC28 protein kinase 2, Spry 2, and ERK 3, were increased with the Z-ratio values of over 2.0 for the cervix cancer tissues compared to those for the normal controls. Those genes were involved In cell growth and proliferation, cell cycle control, or signal transduction. The expressions of the other 6 genes, Including G protein coupled receptor kinase 5, were decreased with the Z-ratio values of below -2.0. After the radiation thorapy, most of the genes, with a previously Increase expressions, represented the decreased expression profiles, and the genes, with the Z-ratio values of over 2.0, were cyclic nucleotlde gated channel and 3 Expressed sequence tags (EST). In the concurrent chemo-radiation group, the genes involved in cell growth and proliferation, cell cycle control, and signal transduction were shown to have increased expressions compared to the radiation therapy alone group. The expressions of genes involved in anglogenesis (angiopoietln-2), immune reactions (formyl peptide receptor-iike 1), and DNA repair (CAMP phosphodiesterase) were increased, however, the expression of gene involved In apoptosls (death associated protein kinase) was decreased. Conclusion : The different kinds of genes involved in the development and progression of cervical cancer were identified with the CDNA microarray, and the proposed theory is that the proliferation signal stalls with ILK, and is amplified with Spry 2 and MAPK signaling, and the cellular mitoses are Increased with the increased expression oi Cdc 2 and cell division kinases. After the radiation therapy, the expression profiles demonstrated 4he evidence of the decreased cancer cell proliferation. There was no sigificant difference in the morphological findings of cell death between the radiation therapy aione and the chemo-radiation groups In the second time biopsy specimen, however, the gene expression profiles were markedly different, and the mechanism at the molecular level needs further study.

Modeling Brand Equity for Lifestyle Brand Extensions: A Strategic Approach into Generation Y vs. Baby Boomer (생활방식품패확장적품패자산건모(生活方式品牌扩张的品牌资产建模): 침대Y세대화영인조소비자적전략로경(针对Y世代和婴儿潮消费者的战略路径))

  • Kim, Eun-Young;Brandon, Lynn
    • Journal of Global Scholars of Marketing Science
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    • v.20 no.1
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    • pp.35-48
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    • 2010
  • Today, the fashion market challenged by a maturing retail market needs a new paradigm in the "evolution of brand" to improve their comparative advantages. An important issue in fashion marketing is lifestyle brand extension with a specific aim to meet consumers' specific needs for their changing lifestyle. For fashion brand extensions into lifestyle product categories, Gen Y and Baby Boomer are emerging as "prospects"-Baby Boomers who are renovating their lifestyle, and generation Y experiencing changes in their life stage-with demands for buying new products. Therefore, it is imperative that apparel companies pay special attention to the consumer cohort for brand extension to create and manage their brand equity in a new product category. The purposes of this study are to (a) evaluate brand equity between parent and extension brands; (b) identify consumers' perceived marketing elements for brand extension; and (c) estimate a structural equation model for examining causative relationship between marketing elements and brand equity for brand extensions in lifestyle product category including home fashion items for the selected two groups (e.g., Gen Y, and Baby boomer). For theoretical frameworks, this study focused on the traditional marketing 4P's mix to identify what marketing element is more importantly related to brand extension equity for this study. It is assumed that comparable marketing capability can be critical to establish "brand extension equity", leads to successfully entering the new categories. Drawing from the relevant literature, this study developed research hypotheses incorporating brand equity factors and marketing elements by focusing on the selected consumers (e.g., Gen Y, Baby Boomer). In the context of brand extension in the lifestyle products, constructs of brand equity consist of brand awareness/association, brand perceptions (e.g., perceived quality, emotional value) and brand resonance adapted from CBBE factors (Keller, 2001). It is postulated that the marketing elements create brand extension equity in terms of brand awareness/association, brand perceptions by the brand extension into lifestyle products, which in turn influence brand resonance. For data collection, the sample was comprised of Korean female consumers in Gen Y and Baby Boomer consumer categories who have a high demand for lifestyle products due to changing their lifecycles. A total of 651 usable questionnaires were obtained from female consumers of Gen Y (n=326) and Baby Boomer (n=325) in South Korea. Structural and measurement models using a correlation matrix was estimated using LISREL 8.8. Findings indicated that perceived marketing elements for brand extension consisted of three factors: price/store image, product, and advertising. In the model of Gen Y consumers, price/store image had a positive effect on brand equity factors (e.g., brand awareness/association, perceived quality), while product had positive effect on emotional value in the brand extensions; and the brand awareness/association was likely to increase the perceived quality and emotional value, leading to brand resonance for brand extensions in the lifestyle products. In the model of Baby Boomer consumers, price/store image had a positive effect on perceived quality, which created brand resonance of brand extension; and product had a positive effect on perceived quality and emotional value, which leads to brand resonance for brand extension in the lifestyle products. However, advertising was negatively related to brand equity for both groups. This study provides an insight for fashion marketers in developing a successful brand extension strategy, leading to a sustainable competitive advantage. This study complements and extends prior works in the brand extension through critical factors of marketing efforts that affect brand extension success. Findings support a synergy effect on leveraging of fashion brand extensions (Aaker and Keller, 1990; Tauber, 1988; Shine et al., 2007; Pitta and Katsanis, 1995) in conjunction with marketing actions for entering into the new product category. Thus, it is recommended that marketers targeting both Gen Y and Baby Boomer can reduce marketing cost for entering the new product category (e.g., home furnishings) by standardized marketing efforts; fashion marketers can (a) offer extension lines with premium ranges of price; (b) place an emphasis on upscale features of store image positioning by a retail channel (e.g., specialty department store) in Korea, and (c) combine apparel with lifestyle product assortments including innovative style and designer’s limited editions. With respect to brand equity, a key to successful brand extension is consumers’ brand awareness or association that ensures brand identity with new product category. It is imperative for marketers to have knowledge of what contributes to more concrete associations in a market entry into new product categories. For fashion brands, a second key of brand extension can be a "luxury" lifestyle approach into new product categories, in that higher price or store image had impact on perceived quality that established brand resonance. More importantly, this study increases the theoretical understanding of brand extension and suggests directions for marketers as they establish marketing program at Gen Y and Baby Boomers.

Sesquiterpenoids Bioconversion Analysis by Wood Rot Fungi

  • Lee, Su-Yeon;Ryu, Sun-Hwa;Choi, In-Gyu;Kim, Myungkil
    • 한국균학회소식:학술대회논문집
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    • 2016.05a
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    • pp.19-20
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    • 2016
  • Sesquiterpenoids are defined as $C_{15}$ compounds derived from farnesyl pyrophosphate (FPP), and their complex structures are found in the tissue of many diverse plants (Degenhardt et al. 2009). FPP's long chain length and additional double bond enables its conversion to a huge range of mono-, di-, and tri-cyclic structures. A number of cyclic sesquiterpenes with alcohol, aldehyde, and ketone derivatives have key biological and medicinal properties (Fraga 1999). Fungi, such as the wood-rotting Polyporus brumalis, are excellent sources of pharmaceutically interesting natural products such as sesquiterpenoids. In this study, we investigated the biosynthesis of P. brumalis sesquiterpenoids on modified medium. Fungal suspensions of 11 white rot species were inoculated in modified medium containing $C_6H_{12}O_6$, $C_4H_{12}N_2O_6$, $KH_2PO_4$, $MgSO_4$, and $CaCl_2$ for 20 days. Cultivation was stopped by solvent extraction via separation of the mycelium. The metabolites were identified as follows: propionic acid (1), mevalonic acid lactone (2), ${\beta}$-eudesmane (3), and ${\beta}$-eudesmol (4), respectively (Figure 1). The main peaks of ${\beta}$-eudesmane and ${\beta}$-eudesmol, which were indicative of sesquiterpene structures, were consistently detected for 5, 7, 12, and 15 days These results demonstrated the existence of terpene metabolism in the mycelium of P. brumalis. Polyporus spp. are known to generate flavor components such as methyl 2,4-dihydroxy-3,6-dimethyl benzoate; 2-hydroxy-4-methoxy-6-methyl benzoic acid; 3-hydroxy-5-methyl phenol; and 3-methoxy-2,5-dimethyl phenol in submerged cultures (Hoffmann and Esser 1978). Drimanes of sesquiterpenes were reported as metabolites from P. arcularius and shown to exhibit antimicrobial activity against Gram-positive bacteria such as Staphylococcus aureus (Fleck et al. 1996). The main metabolites of P. brumalis, ${\beta}$-Eudesmol and ${\beta}$-eudesmane, were categorized as eudesmane-type sesquiterpene structures. The eudesmane skeleton could be biosynthesized from FPP-derived IPP, and approximately 1,000 structures have been identified in plants as essential oils. The biosynthesis of eudesmol from P. brumalis may thus be an important tool for the production of useful natural compounds as presumed from its identified potent bioactivity in plants. Essential oils comprising eudesmane-type sesquiterpenoids have been previously and extensively researched (Wu et al. 2006). ${\beta}$-Eudesmol is a well-known and important eudesmane alcohol with an anticholinergic effect in the vascular endothelium (Tsuneki et al. 2005). Additionally, recent studies demonstrated that ${\beta}$-eudesmol acts as a channel blocker for nicotinic acetylcholine receptors at the neuromuscular junction, and it can inhibit angiogenesis in vitro and in vivo by blocking the mitogen-activated protein kinase (MAPK) signaling pathway (Seo et al. 2011). Variation of nutrients was conducted to determine an optimum condition for the biosynthesis of sesquiterpenes by P. brumalis. Genes encoding terpene synthases, which are crucial to the terpene synthesis pathway, generally respond to environmental factors such as pH, temperature, and available nutrients (Hoffmeister and Keller 2007, Yu and Keller 2005). Calvo et al. described the effect of major nutrients, carbon and nitrogen, on the synthesis of secondary metabolites (Calvo et al. 2002). P. brumalis did not prefer to synthesize sesquiterpenes under all growth conditions. Results of differences in metabolites observed in P. brumalis grown in PDB and modified medium highlighted the potential effect inorganic sources such as $C_4H_{12}N_2O_6$, $KH_2PO_4$, $MgSO_4$, and $CaCl_2$ on sesquiterpene synthesis. ${\beta}$-eudesmol was apparent during cultivation except for when P. brumalis was grown on $MgSO_4$-free medium. These results demonstrated that $MgSO_4$ can specifically control the biosynthesis of ${\beta}$-eudesmol. Magnesium has been reported as a cofactor that binds to sesquiterpene synthase (Agger et al. 2008). Specifically, the $Mg^{2+}$ ions bind to two conserved metal-binding motifs. These metal ions complex to the substrate pyrophosphate, thereby promoting the ionization of the leaving groups of FPP and resulting in the generation of a highly reactive allylic cation. Effect of magnesium source on the sesquiterpene biosynthesis was also identified via analysis of the concentration of total carbohydrates. Our current study offered further insight that fungal sesquiterpene biosynthesis can be controlled by nutrients. To profile the metabolites of P. brumalis, the cultures were extracted based on the growth curve. Despite metabolites produced during mycelia growth, there was difficulty in detecting significant changes in metabolite production, especially those at low concentrations. These compounds may be of interest in understanding their synthetic mechanisms in P. brumalis. The synthesis of terpene compounds began during the growth phase at day 9. Sesquiterpene synthesis occurred after growth was complete. At day 9, drimenol, farnesol, and mevalonic lactone (or mevalonic acid lactone) were identified. Mevalonic acid lactone is the precursor of the mevalonic pathway, and particularly, it is a precursor for a number of biologically important lipids, including cholesterol hormones (Buckley et al. 2002). Farnesol is the precursor of sesquiterpenoids. Drimenol compounds, bi-cyclic-sesquiterpene alcohols, can be synthesized from trans-trans farnesol via cyclization and rearrangement (Polovinka et al. 1994). They have also been identified in the basidiomycota Lentinus lepideus as secondary metabolites. After 12 days in the growth phase, ${\beta}$-elemene caryophyllene, ${\delta}$-cadiene, and eudesmane were detected with ${\beta}$-eudesmol. The data showed the synthesis of sesquiterpene hydrocarbons with bi-cyclic structures. These compounds can be synthesized from FPP by cyclization. Cyclic terpenoids are synthesized through the formation of a carbon skeleton from linear precursors by terpene cyclase, which is followed by chemical modification by oxidation, reduction, methylation, etc. Sesquiterpene cyclase is a key branch-point enzyme that catalyzes the complex intermolecular cyclization of the linear prenyl diphosphate into cyclic hydrocarbons (Toyomasu et al. 2007). After 20 days in stationary phase, the oxygenated structures eudesmol, elemol, and caryophyllene oxide were detected. Thus, after growth, sesquiterpenes were identified. Per these results, we showed that terpene metabolism in wood-rotting fungi occurs in the stationary phase. We also showed that such metabolism can be controlled by magnesium supplementation in the growth medium. In conclusion, we identified P. brumalis as a wood-rotting fungus that can produce sesquiterpenes. To mechanistically understand eudesmane-type sesquiterpene biosynthesis in P. brumalis, further research into the genes regulating the dynamics of such biosynthesis is warranted.

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A Tool Box to Evaluate the Phased Array Coil Performance Using Retrospective 3D Coil Modeling (3차원 코일 모델링을 통해 위상배열코일 성능을 평가하기 위한 프로그램)

  • Perez, Marlon;Hernandez, Daniel;Michel, Eric;Cho, Min Hyoung;Lee, Soo Yeol
    • Investigative Magnetic Resonance Imaging
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    • v.18 no.2
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    • pp.107-119
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    • 2014
  • Purpose : To efficiently evaluate phased array coil performance using a software tool box with which we can make visual comparison of the sensitivity of every coil element between the real experiment and EM simulation. Materials and Methods: We have developed a $C^{{+}{+}}$- and MATLAB-based software tool called Phased Array Coil Evaluator (PACE). PACE has the following functions: Building 3D models of the coil elements, importing the FDTD simulation results, and visualizing the coil sensitivity of each coil element on the ordinary Cartesian coordinate and the relative coil position coordinate. To build a 3D model of the phased array coil, we used an electromagnetic 3D tracker in a stylus form. After making the 3D model, we imported the 3D model into the FDTD electromagnetic field simulation tool. Results: An accurate comparison between the coil sensitivity simulation and real experiment on the tool box platform has been made through fine matching of the simulation and real experiment with aids of the 3D tracker. In the simulation and experiment, we used a 36-channel helmet-style phased array coil. At the 3D MRI data acquisition using the spoiled gradient echo sequence, we used the uniform cylindrical phantom that had the same geometry as the one in the FDTD simulation. In the tool box, we can conveniently choose the coil element of interest and we can compare the coil sensitivities element-by-element of the phased array coil. Conclusion: We expect the tool box can be greatly used for developing phased array coils of new geometry or for periodic maintenance of phased array coils in a more accurate and consistent manner.

Clinical Benefits and Complications of Cryotherapy in Advanced Lung Cancer with Central Airway Obstruction (중심성 기도 폐쇄를 동반한 폐암에서 냉동치료의 임상적 유용성 및 부작용)

  • Jung, Jin Yong;Lee, Sung Yong;Kim, Dae Hyun;Lee, Kyung Joo;Lee, Eun Joo;Kang, Eun Hae;Jung, Ki Hwan;Kim, Je Hyeong;Shin, Chol;Shim, Jae Jeong;In, Kwang Ho;Kang, Kyung Ho;Yoo, Se Hwa
    • Tuberculosis and Respiratory Diseases
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    • v.64 no.4
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    • pp.272-277
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    • 2008
  • Background: The efficacy of the use of the interventional bronchoscope for palliation of patients with central airway obstruction has been established. In the palliative setting to alleviate central airway obstruction, the use of laser resection, electrocautery, argon plasma coagulation, photodynamic therapy and cryotherapy can provide relief of an airway obstruction. Cryotherapy is the therapeutic application of extreme cold for the local destruction of living tissue. Recently, this technique has been used for endoscopic management of central airway obstructions in Korea. We report the role and complications of the use of cryotherapy for airway obstructions in patients with advanced lung cancer. Methods: We used a flexible cryoprobe for cryotherapy using nitrous oxide as a cryogen. The cryoprobe was applied through the working channel of a flexible fiberoptic bronchoscope. The temperature of the tip was approximately $-89^{\circ}C$, and the icing time was 5~20 seconds. Results: Four patients with a central airway obstruction from advanced lung cancer were treated with cryotherapy. Three of the four patients were treated successfully and the airway obstruction was improved after the cryotherapy procedure. Dyspnea, hypoxia and atelectais were improved in three cases. Two patients experienced complications- one patient experienced pneumomediastinum and the other patient experienced massive hemoptysis during the cryotherapy procedure. However, these complications resolved and did not influence mortality. Conclusion: This technique is effective and relatively safe for palliation of inoperable advanced lung cancer with a central airway obstruction.

Surgical Approaches to the Middle Cranial Base Tumors (중두개와저 종양에 대한 수술적 치료)

  • Kim, Il Seub;Rha, Hyung Kyun;Lee, Kyung Jin;Cho, Kyung Keun;Park, Sung Chan;Park, Hae Kwan;Cho, Jeung Ki;Kang, Jun Ki;Choi, Chang Rhack
    • Journal of Korean Neurosurgical Society
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    • v.30 no.9
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    • pp.1079-1085
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    • 2001
  • Objective : We analysed various surgical approaches and surgical results of 28 middle cranial base tumors for the purpose of selecting optimal surgical approach to the middle cranial base tumor. Methods : In this retrospective review, 28 patients, including 16 meningioma, 6 trigeminal neurinoma, 2 pituitary adenoma, 2 craniopharyngioma, 1 facial neurinoma, and 1 metastatic tumor, underwent surgical treatment using skull base technique. Of theses, 16 tumors were mainly confined to middle cranial fossae, 5 tumors with extension into both anterior and middle fossa, and 7 tumors with extension into both middle and posterior fossa. Tumors that confined to the middle cranial fossa or extended into the anterior cranial fossa were operated with modified pterional, orbitozygomatic or Dolen'c approach, and tumors that extended into the posterior cranial fossa were operated with anterior, posterior or combined transpetrosal approach. Completeness of tumor resection, surgical outcome, postoperative complication, and follow up result were studied. Results : Total tumor removal was achieved in 9 tumors of 10 tumors that did not extended to the cavernous sinus, and was achieved in 7 tumors of 8 tumors that extended to the lateral wall of the cavernous sinus. Of 10 tumors that extended to the venous channel of the cavernous sinus, only 2 were removed totally. Surgical outcome was excellent in 14 patients, good in 10, fair in 2 and poor in 2. There were no death in this series. Dumbell type tumor which extended into both middle and posterior fossae showed tendency of poor prognosis as compared with tumors that confined middle cranial fossa and extended into both anterior and middle cranial fossa. Postoperative dysfunctions were trieminal hypesthesia in 3, oculomotor nerve palsy in 2, abducens nerve palsy in 2, hemiparesis in 2, cerebellar sign in 1, facial palsy in 1 and hearing impairment in 1. Conclusion : Based on our findings and a review of the literature, we conclude that, when selecting the surgical approach to the middle cranial fossa tumors, the most important factors to be considered were exact location of the tumor mass and existence of the cavernous sinus invasion by tumor mass. We recommend modified pterional or orbitozygomatic approach in cases with tumors located anterior and middle cranial base, without cavernous sinus invasion. In cases with tumors invading into cavernous sinus, we recommend Dolen'c or orbitozygomatic approach. And in lateral wall mass and the cavernous sinus, it is preferred to approach the tumor extradurally. For the tumor involing with middle fossa and posterior fossa(dumbell type) a combined petrosal approach is necessary. In cases with cavernous sinus invasion and internal carotid artery encasement, we recommend subtotal resection of the tumor and radiation therapy to prevent permanent postoperative sequele.

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EEG based Cognitive Load Measurement for e-learning Application (이러닝 적용을 위한 뇌파기반 인지부하 측정)

  • Kim, Jun;Song, Ki-Sang
    • Korean Journal of Cognitive Science
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    • v.20 no.2
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    • pp.125-154
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    • 2009
  • This paper describes the possibility of human physiological data, especially brain-wave activity, to detect cognitive overload, a phenomenon that may occur while learner uses an e-learning system. If it is found that cognitive overload to be detectable, providing appropriate feedback to learners may be possible. To illustrate the possibility, while engaging in cognitive activities, cognitive load levels were measured by EEG (electroencephalogram) to seek detection of cognitive overload. The task given to learner was a computerized listening and recall test designed to measure working memory capacity, and the test had four progressively increasing degrees of difficulty. Eight male, right-handed, university students were asked to answer 4 sets of tests and each test took from 61 seconds to 198 seconds. A correction ratio was then calculated and EEG results analyzed. The correction ratio of listening and recall tests were 84.5%, 90.6%, 62.5% and 56.3% respectively, and the degree of difficulty had statistical significance. The data highlighted learner cognitive overload on test level of 3 and 4, the higher level tests. Second, the SEF-95% value was greater on test3 and 4 than on tests 1 and 2 indicating that tests 3 and 4 imposed greater cognitive load on participants. Third, the relative power of EEG gamma wave rapidly increased on the 3rd and $4^{th}$ test, and signals from channel F3, F4, C4, F7, and F8 showed statistically significance. These five channels are surrounding the brain's Broca area, and from a brain mapping analysis it was found that F8, right-half of the brain area, was activated relative to the degree of difficulty. Lastly, cross relation analysis showed greater increasing in synchronization at test3 and $4^{th}$ at test1 and 2. From these findings, it is possible to measure brain cognitive load level and cognitive over load via brain activity, which may provide atimely feedback scheme for e-learning systems.

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Characterization of Trigeminal Ganglion Neurons Expressing Transient Receptor Potential Ankyrin 1 (TRPA1) in the Rat (흰쥐의 삼차신경절에서 Transient receptor potential ankyrin 1 (TRPA1)의 발현 특성에 관한 연구)

  • Paik, Sang-Kyoo;Na, Yeon-Kyung;Kim, Yun-Sook
    • Applied Microscopy
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    • v.42 no.1
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    • pp.27-33
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    • 2012
  • Transient receptor potential ankyrin 1 (TRPA1), responding to noxious cold (${\leq}17^{\circ}C$) and pungent compounds, is implicated in nociception, but little is known about the coexpression of TRPA1 and other channels or receptors involved in the nociception in craniofacial regions. To address this issue, we characterized the TRPA1-immunopositive (+) neurons in the rat trigeminal ganglion (TG) and investigated their colocalization with other proteins known to be expressed in nociceptive neurons, such as transient receptor potential vanilloid (TRPV1) and $P2X_3$ receptor, using light microscopic immunofluorescence labeling method with TRPA1 and TRPV1 or $P2X_3$ antisera. The majority of TRPA1+ neurons costained for TRPV1 (TRPV1+/TRPA1+; 58.8%, 328/558) and 41.2% only expressed TRPA1 but not TRPV1. The TRPV1+/TRPA1+ neurons were small and medium sized. In addition, we investigated the colocalization of TRPA1 with $P2X_3$, a nonselective cation channel activated by ATP that may be released in the extracellular space as a result of tissue damage and inflammation. Among all TRPA1+ TG neurons, 26.1% (310/1186) costained for $P2X_3$, whereas 73.9% (876/1186) of TRPA1+ neurons did not coexpress $P2X_3$. $P2X_3$+/TRPA1+ neurons were predominantly small and medium sized. These results suggest that TRPA1+ neurons coexpressing TRPV1 or $P2X_3$ are involved in specific roles in the transmission and processing of orofacial nociceptive information by noxious cold, heat, and inflammation.

Expression and Purification of the Phosphatase-like Domain of a Voltage-Sensing Phosphatase, Ci-VSP (막 전위 감지 탈인산화 효소, Ci-VSP의 유사 탈인산화 효소 도메인의 발현과 정제)

  • Kim, Sung-Jae;Kim, Hae-Min;Choi, Hoon;Kim, Young-Jun
    • Journal of Life Science
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    • v.21 no.7
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    • pp.1032-1038
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    • 2011
  • Recently identified Ciona intestinalis voltage sensor-containing phosphatase (Ci-VSP) consists of an ion channel-like transmembrane domain (VSD) and a phosphatase-like domain. Ci-VSP senses the change of membrane potential by its VSD and works as a phosphoinositide phosphatase by its phosphatase domain. In this study, we present the construction of His-tagged phosphatase-like domain of Ci-VSP, its recombinant expression and purification, and its enzymatic activity behavior in order to examine the biochemical behavior of phosphatase domain of Ci-VSP without interference. We found that Ci-VSP(248-576)-His can be eluted with an elution buffer containing 25 mM NaCl and 100 mM imidazole during His-tag purification. In addition, we found the proper measurement condition for kinetics study of Ci-VSP(248-576)-His against p-nitrophenyl phosphate (pNPP). We measured the kinetic constant of Ci-VSP(248-576)-His at $37^{\circ}C$, pH 5.0 or 5.5, under 30 min of reaction time, and less than $2.0\;{\mu}g$ of protein amount. With these conditions, we acquired that Ci-VSP(248-576)-His has $K_m$ of $354{\pm}0.143\;{\mu}M$, $V_{max}$ of $0.0607{\pm}0.0137\;{\mu}mol$/min/mg and $k_{cat}$ of $0.359{\pm}0.009751\;min^{-1}$ for pNPP dephosphorylation. Therefore, we produced a pure form of Ci-VSP(248-576)-His, and this showed a higher activity against pNPP. This purified protein will provide the road to a structural investigation on an interesting protein, Ci-VSP.

Health Condition Assessment Using the Riparian Vegetation Index and Vegetation Analysis of Geumgang mainstream and Mihocheon (수변식생지수를 이용한 금강본류와 미호천의 건강성 평가 및 식생분석)

  • Lee, Seung-Yeon;Jang, Rae-Ha;Han, Young-Sub;Jung, Young-Ho;Lee, Soo-In;Lee, Eung-Pill;You, Young-Han
    • Korean Journal of Environment and Ecology
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    • v.32 no.1
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    • pp.105-117
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    • 2018
  • This study conducted health assessment and multivariate vegetation analysis using the riparian vegetation index in 30 sites of the Geumgang mainstream and Mihocheon to obtain practical data on the river management of the Geumgang. The result showed that the number of plant communities was 54. The flora was 75 families, 185 genera, 243 species, 2 subspecies, 21 varieties, 2 varieties, and 268 taxa. The riparian vegetation index was 38.3 (3.3; G-D1 ~ 66.7; G-U2, G-U4, and G-M3), and the health of the rivers in this area was evaluated as normal (grade C). The health of rivers was the highest in the upper stream of Geumgang mainstream and lowest in the downstream of Geumgang mainstream. The relationship between riparian vegetation index and chlorophyll-a content was low. The riparian vegetation was divided into five groups of Digitaria ciliaris colony group, Salix gracilistyla colony group, Erigeron annuus colony group, the group dominated by Humulus japonicus, Salix koreensis, Miscanthus sacchariflorus, and Phragmites japonica colonies, and the group dominated by Conyza canadensis and Echinochloa crusgalli var. echinata colonies. They had the similar health conditions. The CCA analysis showed that the environmental factors affecting the distribution of vegetation were physical factors such as vegetation area, artificial structure area, waterway area, branch width, channel width, and bank height and the biological factors such as the number of species. As such, it is necessary to maintain the health condition through continuous monitoring where the health condition is high and to apply active measures such as ecological restoration where the health condition is low.