• Title/Summary/Keyword: 3-D Mechanism

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PKA-Mediated Regulation of B/K Gene Transcription in PC12 Cells

  • Choi, Mi-Hyun;Kim, Ho-Shik;Choi, Sung-Ho;Kim, Mi-Young;Jang, Yoon-Seong;Jang, Young-Min;Lee, Jeong-Hwa;Jeong, Seong-Whan;Kim, In-Kyung;Kwon, Oh-Joo
    • The Korean Journal of Physiology and Pharmacology
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    • v.9 no.6
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    • pp.333-339
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    • 2005
  • B/K protein is a novel protein containing double C2-like domains. We examined the specific signaling pathway that regulates the transcription of B/K in PC12 cells. When the cells were treated with forskolin ($50{\mu}M$), B/K mRNA and protein levels were time-dependently decreased, reaching the lowest level at 3 or 4 hr, and thereafter returning to the control level. Chemicals such as dibutyryl-cAMP, cellpermeable cyclic AMP (cAMP) analogue and CGS21680, adenosine receptor $A_{2A}$ agonist, also repressed the B/K transcription. However, 1,9-dideoxyforskolin did not show inhibitory effect on B/K transcription, suggesting direct involvement of cAMP in the forskolin-induced inhibition of B/K transcription. Effect of forskolin, dibutyryl cAMP and CGS21680 was significantly reduced in PKA-deficient PC12 cell line (PC12-123.7). One cAMP-response element (CRE)-like sequence (B/K CLS) was found in the promoter region of B/K DNA, and electrophoretic mobility shift assay indicated its binding to CREM and CREB. Forskolin significantly suppressed the promoter activity in CHO-K1 cells transfected with the constructs containing B/K CLS, but not with the construct in which B/K CLS was mutated (AC:TG). Taken together, we suggest that the transcription of B/K gene in PC12 cells may be regulated by PKA-dependent mechanism.

Three Non-Aspartate Amino Acid Mutations in the ComA Response Regulator Receiver Motif Severely Decrease Surfactin Production, Competence Development, and Spore Formation in Bacillus subtilis

  • Wang, Xiaoyu;Luo, Chuping;Liu, Youzhou;Nie, Yafeng;Liu, Yongfeng;Zhang, Rongsheng;Chen, Zhiyi
    • Journal of Microbiology and Biotechnology
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    • v.20 no.2
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    • pp.301-310
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    • 2010
  • Bacillus subtilis strains produce a broad spectrum of bioactive peptides. The lipopeptide surfactin belongs to one well-known class, which includes amphiphilic membrane-active biosurfactants and peptide antibiotics. Both the srfA promoter and the ComP-ComA signal transduction system are an important part of the factor that results in the production of surfactin. Bs-M49, obtained by means of low-energy ion implantation in wild-type Bs-916, produced significantly lower levels of surfactin, and had no obvious effects against R. solani. Occasionally, we found strain Bs-M49 decreased spore formation and the development of competence. Blast comparison of the sequences from Bs-916 and M49 indicate that there is no difference in the srfA operon promoter PsrfA, but there are differences in the coding sequence of the comA gene. These differences result in three missense mutations within the M49 ComA protein. RT-PCR analyses results showed that the expression levels of selected genes involved in competence and sporulation in both the wild-type Bs-916 and mutant M49 strains were significantly different. When we integrated the comA ORF into the chromosome of M49 at the amyE locus, M49 restored hemolytic activity and antifungal activity. Then, HPLC analyses results also showed the comA-complemented strain had a similar ability to produce surf actin with wild-type strain Bs-916. These data suggested that the mutation of three key amino acids in ComA greatly affected the biological activity of Bacillus subtilis. ComA protein 3D structure prediction and motif search prediction indicated that ComA has two obvious motifs common to response regulator proteins, which are the N-terminal response regulator receiver motif and the C-terminal helix-turn-helix motif. The three residues in the ComA N-terminal portion may be involved in phosphorylation activation mechanism. These structural prediction results implicate that three mutated residues in the ComA protein may play an important role in the formation of a salt-bridge to the phosphoryl group keeping active conformation to subsequent regulation of the expression of downstream genes.

MICROSATELLITE INSTABILITY AND p53 GENE MUTATION IN ORAL SQUAMOUS CELL CARCINOMA (구강편평상피세포암 조직에서의 Microsatellite 불안정성 및 p53 유전자 돌연변이 분석)

  • Choi, Tae-Ho;Chung, Woon-Bok;Hong, Su-Hyung;Kim, Jin-A;Na, Sun-Oung;Jang, Hyun-Jung;Sohn, Yoon-Kyung;Kim, Chin-Soo;Kim, Jung-Wan
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.26 no.4
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    • pp.337-344
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    • 2000
  • Germ-line mutations at DNA repair loci confer susceptibility to colon cancer in hereditary non-polypopsis colorectal cancer. Somatic loss of DNA mismatch repair gene has been reported in a large variety of other tumor types. Replication errors(RERs) judged by microsatellite instability(MSI) and its associated mutations have been recognized as an important mechanism in various tumor types. To investigate associations between MSI and oral squamous cell carcinoma, the frequency of MSI using 12 microsatellite markers were analyzed for the series of oral tumors. Of 17 tumors, 8 cases(47%) did not show instability at any of the 12 loci; 5(29%) showed instability at $2{\sim}3$ loci; and 4(24%) showed instability above 4 loci. The 4 cases showing widespread MSI did not differ from those without evidence of instability in terms of age at diagnosis, degree of differentiation, metastasis to lymph node, tumor location or the presence of mutations in the p53 tumor suppressor gene. DCC and D17S 796 were the most frequently detected in MSI analysis. There were no correlation between smoking and MSI frequency, instead, smoking was suggested to increase the mutation rate of p53 and development of oral carcinomas.

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Effects of Photoperiod and Water Temperature on the Reproductive cycle of the Spring-Spawning Bitterling, Rhodeus uyekii (Pisces : Cyprinidae) (각시붕어, Rhodeus uyekii의 생식주기에 미치는 광주기 및 수온의 영향)

  • An, Cheul-Min
    • Korean Journal of Ichthyology
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    • v.7 no.1
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    • pp.43-55
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    • 1995
  • Based on the reproductive cycle of Rhodeus uyekii, the reproductive control mechanism was examined under the several combinations of photoperiod and temperature regimes at different phases of their reproductive cycle. In early spring, the gonads developed rapidly under the warm temperature condition(above $8^{\circ}C$), regardless of the photoperiod. In late spring or early summer, gonads intensely regressed under the high temperature condition(above $24^{\circ}C$), regardless of the photoperiod. Thus, it is concluded that the spawning period of R. uyekii is initiated by the rising of water temperature in spring and is terminated by the high temperature in early summer. In autumn, the gonadal recrudescence occurs under the conditions of artificial long daylength with warm temperature(15L/9D, $12^{\circ}C{\sim}20^{\circ}C$), whereas the gonads remain in the existing condition under the short daylength. Therefore, in autumn the gonadal recrudescence is prevented by the short daylength although the temperature is still in favorable condition. In bitterling, the responsiveness of the gonads to photoperiod varies clearly with seasons, which was declined during the winter but increased during the autumn. The critical photoperiod for maturation ranges 12~13 hours of light per day. The results indicate that the proper ranges of temperature and photoperiod for gonadal maturation of this species are from $8^{\circ}C$ to $8^{\circ}C$ and from 13L to 15L, respectively.

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Salinity Effects on the Hydraulic Conductivity of Uplands (밭토양(土壌)의 수리전도도(水理伝導度)에 대(対)한 염류효과(塩類効果))

  • Park, Chang-Seo;O'Connor, George A.
    • Korean Journal of Soil Science and Fertilizer
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    • v.16 no.1
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    • pp.7-13
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    • 1983
  • Laboratory determinations of saturated hydraulic conductivity were conducted with four soils varying in texture from sand to clay and with five waters with different salinity level. The waters varied in total dissolved solids from 1,250 to $15,000mg/{\ell}$ and in SAR from 16 to 57 and were representative of saline waters in New Mexico. Saturated hydraulic conductivities of the soils were not significantly affected by water salinity if these waters were the sole source of irrigation water. However, small additions of distilled water, assuming simulated to rain, to soils previously equilibrated with the saline waters significantly decreased soil permeability. Dispersion and short or long-distance transport of clay apparently clogged conducting pores when distilled water was introduced. Swelling was an important mechanism in reducing soil permeability only in the clay soil. The data suggest that, when saline water is the dominant irrigation source and is supplemented by rain, (1) all saline waters could be used on very sandy soils, (2) no saline waters should be used on very heavy soils, and (3) slightly saline, but not very saline, waters could be used on medium-textured soils.

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The Processing of Causative and Passive Verbs in Korean (한국어의 사.피동문 처리에 관한 연구:실어증 환자의 처리 양상을 바탕으로)

  • 문영선;김동휘;남기춘
    • Proceedings of the Korean Society for Cognitive Science Conference
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    • 2000.05a
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    • pp.267-272
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    • 2000
  • 본 연구에서는 한국어의 사·피동문을 실어증 환자가 처리하는 양상에 대하여 살펴보았다. 한국어의 사·피동문은 용언에 파생접사가 붙어 이루어지는 경우와 '-게 하다'나 '-어 지다'와 같이 구문 변형으로 하여, 실어증 환자에게 실험을 하였다. 실험에 참여한 환자는 명칭성 실어증 환자, 이해성 실어증 환자, 표현성 실어증 환자, 전반성 실어증 환자로 구성되어 있다. 본 실험에서는 단어 채워 넣기 과제(word completion task)를 사용하였다. 명칭성 실어증 환자의 경우 피동에서는 처리 오류를 보이는 반면, 사동에는 아무런 문제도 보이지 않았다. 표현성 실어증 환자의 경우, <피동-비변형>에서 오류를 많이 보였다. 이를 통해 한국어의 사·피동은 영어와 달리 통사상의 문제가 아니라는 결론을 내릴 수 있다. 즉 이미 사·피동 접사에 의해 파생된 단어가 어휘부에 저장되어 있고, 각 단어의 논항 정보에 따라 문장이 생성되는 것이다. 표현성 실어증 환자가 피동의 비변형에서 지배적인 오류를 보이는 것은 피동의 비변형이 타동사로서 변형인 피동형에 비해 하나의 논항을 더 취하기 때문이다. 이해성 실어증 환자의 경우 사·피동 생성에 별 어려움을 보이지 않았다. 이는 이해성 실어증 환자가 개별 어휘의 논항 정보에 손실을 적게 입고 있음을 시사하는 결과이다. 본 연구에서는 서로 다른 유형을 보이는 환자들을 대상으로 한국어의 사·피동의 처리양상을 대조한 결과, 첫재 사·피동은 서로 다른 통사, 의미상의 처리 양상을 보이고 있고, 둘째 파생접사가 결합된 형태로 어휘부에 저장되어 있는 개별 사·피동사에 의해 형성되는 것임을 확인하였다.d CO2 quantity causes flame temperature to fall since at high strain retes diluent effect is prevailing and at low strain rates the products inhibits chain branching. It is also found that the contribution of NO production by N2O and NO2 mechanisms are negligible and that thermal mechanism is concentrate on only the reaction zone. As strain rate and CO2 quantity increase, NO production is remarkably augmented.our 10%를 대용한 것이 무첨가한 것보다 많이 단단해졌음을 알 수 있었다. 혼합중의 반죽의 조사형 전자현미경 관찰로 amarans flour로 대체한 gluten이 단단해졌음을 알수 있었다. 유화제 stearly 칼슘, 혹은 hemicellulase를 amarans 10% 대체한 밀가루에 첨가하면 확연히 비용적을 증대시킬 수 있다는 사실을 알 수 있었다. quinoa는 명아주과 Chenopodium에 속하고 페루, 볼리비아 등의 고산지에서 재배 되어지는 것을 시료로 사용하였다. quinoa 분말은 중량의 5-20%을 quinoa를 대체하고 더욱이 분말중량에 대하여 0-200ppm의 lipase를 lipid(밀가루의 2-3배)에 대하여 품질개량제로서 이용했다.

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Predictive Factors of Blood Transfusion Requirement in Blunt Trauma Patients Admitted to the Emergency Room (응급실에 내원한 둔상환자의 수혈 필요성 예측인자)

  • Oh, Ji Sun;Kim, Hyung Min;Choi, Se Min;Choi, Kyoung Ho;Hong, Tae Yong;Park, Kyu Nam;So, Byung Hak
    • Journal of Trauma and Injury
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    • v.22 no.2
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    • pp.218-226
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    • 2009
  • Purpose: In multiple blunt trauma patients, transfusion may be a significant therapeutic adjunct to non-operative management. The blood products must be expedited and efficiently to patients in impending shock caused by hemorrhage or traumatic coagulopathy, but the decision to perform blood transfusion has been made empirically, based on the clinician' and has not been guided by objective parameters, but own opinion, that may result in an underestimate of or a failure to detect bleeding, in delayed transfusion, and in a reduced outcome. This article presents quickly assessable predictive factors for determining if a blood transfusion is required to improve outcomes in multiple blunt trauma patients admitted to the emergency room. Methods: In a retrospective review of 282 multiple blunt trauma patients who visited our emergency center by emergency rescuer during a 1-year period, possible factors predictive of the need for a blood transfusion were subjected to univariate and multivariate logistic regression analysis. Results: Of blunt trauma patients, 9.2% (26/282), received red blood cells in the first 24 hours of care. Univariate analysis revealed significant associations between blood transfused and heart rate (HR) > 100 beats/min, respiratory rate (RR) > 20 breaths/min, Glasgow Coma Scale (GCS) < 14, Revised Trauma Score (RTS) < 11, white blood cell count (WBC) < 4000 or > 10000, and initial abnormal portable trauma series (Cspine lateral, chest AP, pelvis AP). A multiple regression analysis, with a correction for diagnosis, identified HR > 100 beats/min (EXP 3.2), GCS < 14 (EXP 4.1), and abnormal trauma series (EXP 2.9), as independent predictors. Conclusion: In our study, systolic blood pressure (SBP) < 90 mmHg, old age > 65 years, hemoglobin < 13g/dL, mechanism of injury were poor predictors of early blood transfusion. Initial abnormal portable trauma series, HR > 100 beats/min, and GCS < 14 were quickly assessable useful factors for predicting a need for early blood transfusion in blunt trauma patients visiting the emergency room.

Platelet-Rich Plasma: Quantitative Assessment of Growth Factor Levels and Comparative Analysis of Activated and Inactivated Groups

  • Lee, Jeong Woo;Kwon, O Hyun;Kim, Taek Kyun;Cho, Young Kyoo;Choi, Kang Young;Chung, Ho Yun;Cho, Byung Chae;Yang, Jung Dug;Shin, Jun Ho
    • Archives of Plastic Surgery
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    • v.40 no.5
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    • pp.530-535
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    • 2013
  • Background Platelet-rich plasma (PRP) has more concentrated platelets than normal plasma (approximately 150-400${\times}10^3$ cell/dL). Platelets excrete several growth factors and cytokines that are associated with the healing and regeneration process. However, even though PRP is widely used, the mechanism or actual effect is presently unclear. Therefore, this study was performed to investigate the levels of growth factors and platelet concentration rate. Methods Autologous blood for preparing PRP was obtained from healthy subjects aged 25 to 35 years. The samples were divided into 4 experimental groups (inactivated whole blood, inactivated PRP, activated whole blood with thrombin and calcium chloride, and activated PRP). The platelet counts in the blood were analyzed and the growth factors were quantitatively measured. A statistical analysis was performed by using Dunn's multiple comparison test. Results In the blood cell analysis, the platelet count of the PRP group was approximately 4.25 times higher than that of the whole blood group. In the quantitative analysis of growth factors, the platelet-derived growth factor (PDGF)-AB, PDGF-BB, and transforming growth factor-${\beta}$ of the inactivated and activated PRP groups were higher than those of the inactivated and activated whole blood groups (P<0.05). Conclusions In this study, the platelet count and the levels of PDGF-AB and PDGF-BB in the PRP were determined. Further, more research is required on the bioactivity level of the growth factors secreted during the process of PRP preparation and the potency of growth factors that can be exerted physiologically in vivo.

Effect of Epidermal Growth Factor (EGF) and anti-EGF on Early Embryonic Development in Mice (Epidermal Growth Factor (EGF)와 anti-EGF가 생쥐배아의 발생에 미치는 영향)

  • 변혜경;이호준
    • Korean Journal of Animal Reproduction
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    • v.21 no.1
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    • pp.61-69
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    • 1997
  • The present study was carried out to investigate the effects of EGF and anti-EGF on early embryonic development and hatching in mice. Developmental and hatching rates of mouse em-bryos from 2-cell to morular stage which were cultured in Ham's FlO medium supplemented with EGF (1-1,000 ng/ml) or anti-EGF (whole serum diluted from 1:10 to 1:1,000) were compared to those of control When mouse early 2-cell embryos were cultured in the EGF supplemented medium, blastulation was accelerated compared with control. Hatching rate was also significantly (p

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Functional Analysis of an Antibiotic Regulatory Gene, afsR2 in S. lividans through DNA microarray System (DNA 마이크로어레이 시스템 분석을 통한 S. lividans 유래 항생제 조절유전자 afsR2 기능 분석)

  • Kim, Chang-Young;Noh, Jun-Hee;Lee, Han-Na;Kim, Eung-Soo
    • KSBB Journal
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    • v.24 no.3
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    • pp.259-266
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    • 2009
  • AfsR2 in Streptomyces lividans, a 63-amino acid protein with limited sequence homology to Streptomyces sigma factors, has been known for a global regulatory protein stimulating multiple antibiotic biosynthetic pathways. Although the detailed regulatory mechanism of AfsK-AfsR-AfsR2 system has been well characterized, very little information about the AfsR2-dependent down-stream regulatory genes were characterized. Recently, the null mutant of afsS in S. coelicolor (the identical ortholog of afsR2) has been characterized through DNA microarray system, revealing that afsS deletion regulated several genes involved in antibiotic biosynthesis as well as phosphate-starvation. Through comparative DNA microarray analysis of afsR2-overexpressed S. lividans, here we also identify several afsR2-dependent genes involved in phosphate starvation, morphological differentiation, and antibiotic regulation in S. lividans, confirming that the AfsR2 plays an important pleiotrophic regulatory role in Streptomyces species.