• 제목/요약/키워드: $P_{2Y}$ receptor

검색결과 1,284건 처리시간 0.03초

랫드 후각점막내 Zinc 이온의 조직화학적 동정 (Histochemical Detection of Ionic Zinc in the Rat Olfactory Mucosa: Zinc Selenium Autometallography ($ZnSe^{AMG}$))

  • 남동우;손원재;김성주;김용국;김수진;유윤조;정영길;조승묵
    • Applied Microscopy
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    • 제33권2호
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    • pp.145-154
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    • 2003
  • 본 연구는 후각점막에 내재하는 zinc의 분포를 형태학적으로 뚜렷이 보여준 예가 없기에 조직화학적으로 염색한 후 광학 및 전자현미경으로 이들의 분포를 기술하고자 하였다. 실험동물로는 성숙한 수컷 랫드(SD 계통)를 사용하였다. 우선 동물은 전신마취시킨 후 만들어진 selenium 용액 $100{\mu}l$를 복강에 주사하였고(i.p 실험군), 다른 방법으로는 selenium 용액 $100{\mu}l$를 비강 깁숙히 플라스틱관을 삽입한 후 Hamilton 주사기로 한 방울씩 떨어뜨렸다(i.n 실험군). 후각점막내 zinc를 조직화학법으로 동정하기 위해서 zinc 특이성이 높은 AMG법(Danscher, 1985)을 이용하였다. 각 실험군의 동물은 주사 후 2시간에 이르러 3% Glutaraldehyde 고정액으로 관류함으로써 희생시켰다. 고정후 비중격을 포함하여 후각부위를 떼어낸 수 $30{\mu}l$ 두께의 관상절편을 만들었다. AMG 반응이 끝나면 toluene blue (TB)로 대조염색하고, 알코올과 자일렌 등을 이용한 탈수과정 및 청명과정을 거쳐 광학현미경하에서 관찰하였다. 전자현미경적 절편을 만들기 위해 전자현미경 관찰을 위한 실험으로 vibratome을 사용하여, $100{\mu}m$ 두께의 가로절편을 만들었고, AMG염색이 끝나면 일반적인 전자현미경적 관찰을 위한 일련의 과정을 거쳐 투과전자현미경하에서 관찰하였다. Selenium 용액을 복강에 투여한 i.p 실험군에서는 $30{\mu}m$ 두께의 후각점막의 상층부와 기저부에서 강한 AMG 반응산물(silver grains)이 관찰되었으나 기저막아래 고유판에서는 전혀 관찰되지 않았다. 반면 selenium 용액을 비강점막에 직접 도포한 i.n 실험군에서는 i.p 실험군과 같이 후각점막에서 강한 AMG 반응이 관찰되었을 뿐만 아니라 이러한 반응이 기저막내 후각실의 주행을 따라 관찰되었다. 투과전자현미경으로 관찰된 후각상피에서는 AMG 과립이 지지세포의 세포 상층부에서 관찰되는 분비과립에 국한되는 소견이며, 이곳에서 관찰된 AMG은 과립(silver grains)은 원형 또는 난원형으로 그 크기는 다양하였다. 반면 세포체 하부에서 관찰된 AMG grains은 주로 용해소체위에서 밀집되어 관찰되었으며, 핵주변부 및 세포사이공간(intercellular space)에서는 AMG grains이 낱개로 구분, 관찰되었다. 한편 i.n 실험군에서 관찰된 후각점막의 전반적인 구조가 손상된 소견을 보였으며, 지지세포의 위쪽에서는 전자밀도가 높고 간상의 crystalloid structure가 다수 관찰되었고, 이들 구조에 다수의 AMG 과립이 붙어 있었다. 본 연구의 결과를 요약하면 랫드 후각점막에 존재하는 zinc는 지지세포의 분비과립과 후각세포의 축삭다발인 후각실에서 관찰되었는데 이는 zinc가 후각기능과 매우 밀접한 관련있음을 시사하며, 이는 향후 후각기능과 zinc의 연관성을 연구하는데 소중한 자료가 될 것으로 믿는다. 또한 본 연구의 결과는 후각점막의 이상으로 나타나는 여러 가지 질환의 병리에서 zinc가 영위하는 신경생물학적 기능을 밝히는데 유용한 기초자료가 될 수 있을 것으로 기대한다.

Molecular Cloning and mRNA Expression of the Porcine Insulin-responsive Glucose Transporter (GLUT4)

  • Zuo, Jianjun;Dai, Fawen;Feng, Dingyuan;Cao, Qingyun;Ye, Hui;Dong, Zemin;Xia, Weiguang
    • Asian-Australasian Journal of Animal Sciences
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    • 제23권5호
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    • pp.640-648
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    • 2010
  • Insulin-responsive glucose transporter 4 (GLUT4) is a member of the glucose transporter family and mainly presents in skeletal muscle and adipose tissue. To clarify the molecular structure of porcine GLUT4, RACE was used to clone its cDNA. Several cDNA clones corresponding to different regions of GLUT4 were obtained by amplifying reverse-transcriptase products of total RNA extracted from Landrace porcine skeletal muscles. Nucleotide sequence analysis of the cDNA clones revealed that porcine GLUT4 cDNA was composed of 2,491 base pairs with a coding region of 509 amino acids. The deduced amino acid sequence was over 90% identical to human, rabbit and cattle GLUT4. The tissue distribution of GLUT4 was also examined by Real-time RT-PCR. The mRNA expression abundance of GLUT4 was heart>liver, skeletal muscle and brain>lung, kidney and intestine. The developmental expression of GLUT4 and insulin receptor (IR) was also examined by Real-time RT-PCR using total RNA extracted from longissimus dorsi (LM), semimembranosus (SM), and semitendinosus (SD) muscle of Landrace at the age of 1, 7, 30, 60 and 90 d. It was shown that there was significant difference in the mRNA expression level of GLUT4 in skeletal muscles of Landrace at different ages (p<0.05). The mRNA expression level of IR also showed significant difference at different ages (p<0.05). The developmental change in the mRNA expression abundance of GLUT4 was similar to that in IR, and both showed a higher level at birth and 30 d than at other ages. However, there was no significant tissue difference in the mRNA expression of GLUT4 or IR (p>0.05). These results showed that the nucleotide sequence of the cDNA clones was highly identical with human, rabbit and cattle GLUT4 and the developmental change of GLUT4 mRNA in skeletal muscles was similar to that of IR, suggesting that porcine GLUT4 might be an insulin-responsive glucose transporter. Moreover, the tissue distribution of GLUT4 mRNA showed that GLUT4 might be an important nutritional transporter in porcine skeletal muscles.

Association between Laryngeal Squamous Cell Carcinoma and Polymorphisms in Tumor Necrosis Factor Related Apoptosis Induce Ligand (TRAIL), TRAIL Receptor and sTRAIL Levels

  • Verim, Aysegul;Turan, Saime;Farooqi, Ammad Ahmad;Kahraman, Ozlem Timirci;Tepe-Karaca, Cigdem;Yildiz, Yemliha;Naiboglu, Baris;Ozkan, Nazli Ezgi;Ergen, Arzu;Isitmangil, Gulbu Aydinoglu;Yaylim, Ilhan
    • Asian Pacific Journal of Cancer Prevention
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    • 제15권24호
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    • pp.10697-10703
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    • 2015
  • The laryngeal squamous cell carcinoma (LSCC) is one of the most common malignant tumors occurring in the head and neck. Tumor necrosis factor related apoptosis induce ligand (TRAIL) and TRAIL-receptors (DR4, DR5, DcR1, DcR2) are known as important members of TRAIL-mediated biochemical signaling pathway. Associations between polymorphisms in these genes and clinicopathological characteristics of human laryngeal carcinoma are not well defined. This study therefore aimed to investigate a possible relationship among the TRAIL and TRAIL-DR4 polymorphisms and sTRAIL levels in the risk or progression of LSCC. A total of 99 patients with laryngeal cancer and 120 healthy subjects were enrolled in the study. DR4 C626G and TRAIL 1595 C/T genotypes were determined by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis and sTRAIL levels were measured by ELISA. There were significant differences in the distribution of DR4 C626G genotypes and frequencies of the alleles between laryngeal cancer patients and controls (p<0.001) but not in TRAIL 1595 C/T. We found the increased frequency of the DR4 C626G homozygote CC genotype in patients than in controls (p<0.001). Haplotype analysis revealed that there was also a statistically significant relationship between TRAIL and TRAIL-DR4 polymorphisms and laryngeal cancer. Serum sTRAIL levels in the laryngeal patients with CC genotype who had advanced tumour stage were lower than those of patients with early tumor stage (p=0.014). Our findings suggest that DR4 C626G genotypes and sTRAIL levels might be associated with progression of laryngeal cancer in the Turkish population.

Immunological benefits by ginseng through reciprocal regulation of Th17 and Treg cells during cyclosporine-induced immunosuppression

  • Heo, Seong Beom;Lim, Sun Woo;Jhun, Joo Yeon;Cho, Mi La;Chung, Byung Ha;Yang, Chul Woo
    • Journal of Ginseng Research
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    • 제40권1호
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    • pp.18-27
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    • 2016
  • Background: It is not clear whether ginseng affects cyclosporine A (CsA)-induced desirable immunosuppressive action. In this study, we evaluated the immunological influence of combined treatment of ginseng with CsA. Methods: Using CD4+ T cells from mouse spleens stimulated with the T cell receptor (TCR) or allogeneic antigen-presenting cells (APCs), we examined the differentiation of naïve T cells into T helper 1 (Th1), Th2, Th17, and regulatory T cells (Tregs), and their cytokine production during treatment by Korean Red Ginseng extract (KRGE) and/or CsA. The influence of KRGE on the allogeneic T cell response was evaluated by mixed lymphocyte reaction (MLR). We also evaluated whether signal transducer and activator of transcription 3 (STAT3) and STAT5 are implicated in this regulation. Results: Under TCR stimulation, KRGE treatment did not affect the population of CD4+interferon gamma ($IFN{\gamma}$)+ and CD4+interleukin (IL)-4+ cells and their cytokine production compared with CsA alone. Under the Th17-polarizing condition, KRGE significantly reduced the number of CD4+IL-17+ cells and CD4+/phosphorylated STAT3 (p-STAT3)+ cells, but increased the number of CD4+CD25+forkhead box P3 (Foxp3)+ cells and CD4+/p-STAT5+ cells compared with CsA alone. In allogeneic APCs-stimulated CD4+ T cells, KRGE significantly decreased total allogeneic T cell proliferation. Consistent with the effects of TCR stimulation, KRGE reduced the number of CD4+IL-17+ cells and increased the number of CD4+CD25+Foxp3+ cells under the Th17-polarizing condition. Conclusion: KRGE has immunological benefits through the reciprocal regulation of Th17 and Treg cells during CsA-induced immunosuppression.

경피용 프로드럭인 에칠 글리콜레이트의 국소자극 및 피부투과성 (Skin Penetration and Local Irritation of Ethyl Glycolate, a Potential Transdermal Prodrug)

  • 양성운;하용호;김종갑;최영욱
    • 약학회지
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    • 제40권2호
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    • pp.155-162
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    • 1996
  • Hyperkeratinization is a dermatologic disorder, which is due to the increase of corneocyte cohesion force. Glycolic acid, an alpha hydroxy acid(AHA), has been used to breakdown the hyperkeratinization processes. However, it has a problem of skin irritation when applied topically, due to the strong acidity especially in high concentration. A molecular optimization of glycolic acid has been tried to reduce the skin irritation by the way of prodrug formation. Ethyl glycolate was synthesized by the esterification of glycolic acid with ethanol in acidic conditions in the presence of sulfuric acid, and examined under the spectroscopic trials, such as UV, IR, $^1H$-NMR, and GC-MS. The physicochemical and biopharmaceutical properties of the prodrug were also evaluated. Through the toxicological tests of both skin irritation and eye mucous irritation, it has been proved that ethyl glycolate was less irritant than glycolic acid, since the pH value of synthetic prodrug was higher than that of glycolic acid. In the penetration test through nude mouse skin by diffusion cell, ethyl glycolate was continuously hydrolyzed to glycolic acid, which was assayed form the receptor compartment. It was obtained that the penetrated amount of ethyl glycolate was five times higher than that of glycolic acid. These results suggest that ethyl glycolate might be a successful prodrug of glycolic acid to reduce the skin irritation and to increase the skin penetration as well.

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한국산 퉁가리과 (Amblycipitidge) 어류 2종 수염의 조직 학적 구조 (Histological Structure of the Barbels of Liobagrus andersoni and L. obesus (Amblycipitidae: Pices) from Korea)

  • 박인석;설동원;임수연;김치홍;강언종;공용근
    • 한국수산과학회지
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    • 제39권4호
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    • pp.338-343
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    • 2006
  • Histological observations of the barbels of torrent catfish (Amblycipitidae) in Korea are reported. The bullhead torrent catfish (Liobagrus andersoni) and Korean torrent catfish (L. obesus) have eight obvious barbels in two pairs each on the maxilla and mandible. Each barbel is composed of an epidermis, dermis, and central cartilagerod. The epidermis of the maxillary and mandibular barbels of the bullhead torrent catfish is thicker than that of the Korean torrent catfish (P<0.05) and is formed of stratified epithelium that contains many cutaneous taste buds and a few small club cells. There are many taste buds on the upper part of each barbel. The dermis consists of loose connective tissue fibers that enclose blood vessels and bundles of nerve fibers. The barbels of these fish can be categorized into stiff and flexible types and are accessory, feeding, and sensory structures. Our results validate the barbels as gustatory receptor organs for these fishes.

Evaluation of Transferrin-Polyethylenimine Conjugate for Targeted Gene Delivery

  • Lee Kyung Man;Kim In Sook;Lee Yong Bok;Shin Sang Chul;Lee Kang Choon;Oh In Joon
    • Archives of Pharmacal Research
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    • 제28권6호
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    • pp.722-729
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    • 2005
  • With the aim to improve the specificity and to reduce the cytotoxicity of polyethylenimine (PEI), we have synthesized the conjugates of the branched PEI (25 kDa) with transferrin. The trans-ferrin-PEI (TP) conjugates with five compositions were synthesized using periodate oxidation method and confirmed by FT-IR spectroscopy and gel permeation chromatography. The free amine contents of TP conjugates, which were able to condense and deliver DNA, increased as the amount of PEI increased. TP/DNA polyplexes were characterized by measuring gel elec-trophoresis, ethidium bromide fluorescence quenching, particle size and zeta potential of complexes. Complete complexation of the polyplexes was observed above the N/P ratio of 5 in TP/DNA, and above 3 in PEI/DNA, respectively. The zeta potential of the complexes decreased as the amount of transferrin in TP conjugates increased. Transfection efficiency of TP conjugates was evaluated in HeLa cell and Jurkat cell systems. Among the five compositions of TP conjugates, TP-2 system mediated a higher $\beta$-galactosidase gene expression than PEI system in Jurkat cell which was known to express elevated numbers of transferrin receptors. From the results of the cell viability based on MTT assay, TP conjugates showed lower cytotoxicity com-pared with the PEI system. We expect that the TP conjugate can be used efficiently as a non-viral gene delivery vector.

Cyclic tensile stress inhibits Wnt/${\beta}$-catenin signaling in human periodontal ligament cells

  • Kim, Ji-Young;Yang, Daum;Kim, Ha-Neui;Jung, Kyoung-Suk;Chang, Young-Il;Lee, Zang-Hee
    • International Journal of Oral Biology
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    • 제34권2호
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    • pp.53-59
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    • 2009
  • Periodontal ligament (PDL) tissue is a connective tissue that is interposed between the roots of the teeth and the inner wall of the alveolar bone socket. PDL is always exposed to physiologic mechanical force such as masticatory force and PDL cells play important roles during orthodontic tooth movement by synthesizing and secreting different mediators involved in bone remodeling. The Wnt/${\beta}$-catenin signaling pathway was recently shown to play a significant role in the control of bone formation. In the present study, we applied cyclic tensile stress of 20% elongation to cultured human PDL cells and assessed its impact after six days upon components of the Wnt/${\beta}$-catenin signaling pathway. RTPCR analysis showed that Wnt1a, Wnt3a, Wnt10b and the Wnt receptor LRP5 were down-regulated, whereas the Wnt inhibitor DKK1 was up-regulated in response to these stress conditions. In contrast, little change was detected in the mRNA expression of Wnt5a, Wnt7b, Fz1, and LRP6. By western blotting we found decreased expression of the ${\beta}$-catenin and p-GSK-3${\beta}$ proteins. Our results thus show that mechanical stress suppresses the canonical Wnt/${\beta}$-catenin signaling pathway in PDL cells.

Neuroprotective Effects of KC0244, a Glycine Site Antagonist, in a Rat Model of Transient Focal Ischemia

  • Ku, Hee-Jung;Churlmin Seong;Park, No-Sang;Changbae Jin
    • 한국응용약물학회:학술대회논문집
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    • 한국응용약물학회 1998년도 Proceedings of UNESCO-internetwork Cooperative Regional Seminar and Workshop on Bioassay Guided Isolation of Bioactive Substances from Natural Products and Microbial Products
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    • pp.143-143
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    • 1998
  • Antagonists acting at the glycine site of the NMDA receptor have been gaining safer alternatives for stroke therapy because they have few adverse effect competitive and noncompetitive NMDA antagonists. Therefore, the neuroprotect novel glycine site antagonist KC0244 were evaluated in a rat model of transient comparison with GV150526A in a developmental phase. Middle cerebral artery oc was produced by insertion of a silicone-coated 4-0 nylon monofilament to the o in male Sprague-Dawley rats under isoflurane anesthesia. After 90 or 120 min retracted and the ischemic tissue reperfused. In 90-min MCAO model, GV150526A was administered 30 min before MCAO or immediately after MCAO. In 120-min MC KC0244 or GV150526A (10 mg/kg, i.p.) was administered 1 hr before MCAO or imme MCAO. Infarct volume was measured 24 hr after MCAO using the 2,3,5-triphe chloride staining method. In 90-min MCAO model, treatments with GV1505 significantly reduce infarct volume although they tended to slightly reduce cor approximately 19% compared with the nontreated group. In 120-min MCAO model with GV150526A did not either significantly reduce infarct volume although the reduce total infarct volume by approximately 16% compared with the vehicle-tre However, 1-hr preischemic and immediate treatments with KC0244 reduced total i 39 and 30% (corrected total infarct volume by 44 and 32%), respectively, co vehicle-treated control group. The results suggest that KC0244 can provid against transient focal ischemic damage with greater in vivo potency than GV150

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Arsenic의 첨가량에 따른 방사선 검출센서 (a-Se)의 전기적 특성 비교 (The Comparison of Electric Characteristics of Radiation Detective Sensor(a-Se) with changing composition ratio of Arsenic)

  • 석대우;강상식;이동길;김재형;남상희
    • 한국전기전자재료학회:학술대회논문집
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    • 한국전기전자재료학회 2002년도 추계학술대회 논문집 Vol.15
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    • pp.391-394
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    • 2002
  • There has recently been much interest and research in developing digital x-ray systems based on using amorphous selenium(a-Se) photoconductors as the image receptor. The amorphous selenium layer that is currently being studied for use as an x-ray photoconductor is not pure a-Se but rather amorphous selenium alloyed with arsenic. We fabricated samples using the selenium and arsenic alloy with various concentrations of the arsenic. In this work, x-ray photoconductor using amorphous selenium alloyed with arsenic were fabricated with different concentrations of the arsenic (0.1 wt.%, 0.3wt.%, 0.5wt.%, 1wt.%, 1.5wt.%, 3wt.%, 5wt.%). The seven kind of samples was fabricated with a-Se alloyed with arsenic through vacuum thermal evaporation. We also investigate the arsenic concentration dependence on the device performance in radiation detector. The electric characteristics of radiation detector devices with changing additive ratio of the arsenic is performed by measuring the x-ray induced photocurrent and integrating it over time to find the total charge. The thickness of a-Se is $100{\mu}m$. Bias voltages $3V/{\mu}m$, $6V/{\mu}m$$9V/{\mu}m$ are applied at the samples. As results, the net charge of a-Se 0.3% As sample is $526.0pC/mR/cm^2$ at $9V/{\mu}m$ bias. The net charge is decreased as with the increasing additive ratio of arsenic.

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