• Title/Summary/Keyword: $I_c$ degradation

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MicroRNA-206 Protects against Myocardial Ischaemia-Reperfusion Injury in Rats by Targeting Gadd45β

  • Zhai, Changlin;Qian, Qang;Tang, Guanmin;Han, Bingjiang;Hu, Huilin;Yin, Dong;Pan, Haihua;Zhang, Song
    • Molecules and Cells
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    • v.40 no.12
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    • pp.916-924
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    • 2017
  • MicroRNAs are widely involved in the pathogenesis of cardiovascular diseases through regulating gene expression via translational inhibition or degradation of their target mRNAs. Recent studies have indicated a critical role of microRNA-206 in myocardial ischaemia-reperfusion (I/R) injury. However, the function of miR-206 in myocardial I/R injury is currently unclear. The present study was aimed to identify the specific role of miR-206 in myocardial I/R injury and explore the underlying molecular mechanism. Our results revealed that the expression level of miR-206 was significantly decreased both in rat I/R group and H9c2 cells subjected to hypoxia/reoxygenation (H/R) compared with the corresponding control. Overexpression of miR-206 observably decreased infarct size and inhibited the cardiomyocyte apoptosis induced by I/R injury. Furthermore, bioinformatics analysis, luciferase activity and western blot assay proved that $Gadd45{\beta}$ (growth arrest DNA damage-inducible gene $45{\beta}$) was a direct target gene of miR-206. In addition, the expression of pro-apoptotic-related genes, such as p53, Bax and cleaved caspase3, was decreased in association with the down-regulation of $Gadd45{\beta}$. In summary, this study demonstrates that miR-206 could protect against myocardial I/R injury by targeting $Gadd45{\beta}$.

Inhibition of angiotensin converting enzyme increases PKCβI isoform expression via activation of substance P and bradykinin receptors in cultured astrocytes of mice

  • Jae-Gyun Choi;Sheu-Ran Choi;Dong-Wook Kang;Hyun Jin Shin;Miae Lee;Jungmo Hwang;Hyun-Woo Kim
    • Journal of Veterinary Science
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    • v.24 no.2
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    • pp.26.1-26.11
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    • 2023
  • Background: Angiotensin-converting enzyme inhibitor (ACEi) inhibits the catalysis of angiotensin I to angiotensin II and the degradation of substance P (SP) and bradykinin (BK). While the possible relationship between ACEi and SP in nociceptive mice was recently suggested, the effect of ACEi on signal transduction in astrocytes remains unclear. Objectives: This study examined whether ACE inhibition with captopril or enalapril modulates the levels of SP and BK in primary cultured astrocytes and whether this change modulates PKC isoforms (PKCα, PKCβI, and PKCε) expression in cultured astrocytes. Methods: Immunocytochemistry and Western blot analysis were performed to examine the changes in the levels of SP and BK and the expression of the PKC isoforms in primary cultured astrocytes, respectively. Results: The treatment of captopril or enalapril increased the immunoreactivity of SP and BK significantly in glial fibrillary acidic protein-positive cultured astrocytes. These increases were suppressed by a pretreatment with an angiotensin-converting enzyme. In addition, treatment with captopril increased the expression of the PKCβI isoform in cultured astrocytes, while there were no changes in the expression of the PKCα and PKCε isoforms after the captopril treatment. The captopril-induced increased expression of the PKCβI isoform was inhibited by a pretreatment with the neurokinin-1 receptor antagonist, L-733,060, the BK B1 receptor antagonist, R 715, or the BK B2 receptor antagonist, HOE 140. Conclusions: These results suggest that ACE inhibition with captopril or enalapril increases the levels of SP and BK in cultured astrocytes and that the activation of SP and BK receptors mediates the captopril-induced increase in the expression of the PKCβI isoform.

Inulin stimulates NO synthesis via activation of PKC-$\alpha$ and protein tyrosine kinase, resulting in the activation of NF-$textsc{k}$B by IFN-ν-primed RAW 264.7 cells

  • Koo, Hyun-Na;Hong, Seung-Heon;Kim, Hyung-Min
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 2003.11a
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    • pp.78-78
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    • 2003
  • Inulin, an active component of Chicorium intybus root, has been shown to stimulate the growth of bifidobacteria, and inhibit colon carcinogenesis. NO mediates a number of the host-defense functions of activated macrophages, including antimicrobial and tumoricidal activity. We examined the effect of inulin on the synthesis of NO in RAW 264.7 cells. Inulin alone had no effect, whereas inulin with IFN-ν synergistically increased the NO production and inducible NO synthase (iNOS) expression in RAW 264.7 cells. Synergy between IFN-ν and inulin was mainly dependent on inulin-induced TNF-${\alpha}$ secretion. Also, protein kinase C (PKC)-${\alpha}$ was involved in the inulin-induced NO production. Inulin-mediated NO production was inhibited by the protein tyrosine kinase (PTK) inhibitor, tyrphostin AG126. Since iNOS gene transcriptions have been shown to be under the control of the NF -$\kappa$B/Rel family of transcription factors, we assessed the effect of inulin on NF -$\kappa$B/Rel using an EMSA. Inulin produced strong induction of NF-$\kappa$B/Rel binding, whereas AP-l binding was slightly induced in RAW 264.7 cells. Inulin stimulated phosphorylation and degradation of I$\kappa$B-${\alpha}$. These results suggest that in IFN-ν-primed RAW 264.7 cells inulin might stimulate NO synthesis via activation of PKC-${\alpha}$ and PTK, resulting in the activation of NF-$\kappa$B.

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Isolation and Identification of the Crude Oil-degrading Psychrotrophic Bacterium and the Characteristics of OCT Plasmid (저온성 원유분해 세균의 분리동정 및 OCT 프라스미드 특성)

  • 김상진;윤희정
    • Microbiology and Biotechnology Letters
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    • v.21 no.1
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    • pp.66-73
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    • 1993
  • Psychrotrophic bacterial strains utilizing crude oil as their sole carbon and energy sources were isolated from Antarctic soil and sea sediments. One of the strains named AI-I showed the hightest activity for emulsification of crude oil and the best growth. This strain was identified as Acinetobacter calcoaceticus. A. calcoaceticus AI-I strain contains a plasmid (OCT plasmid) which was related to the utilization of alkane compounds. The molecular weight of this plasmid was estimated to be about 110 Md by agarose gel electrophoresis. The cured strain of A. calcoaceticus AI-I strain (OCT ) was not able to utilize normal hydrocarbon compounds ($C_6C_{17}$) as carbon and energy sources. A. ca/coaceticus AI-1 was resistant to ampicillin and sensitive to streptomycin, kanamycin, chloramphenicol, tetracycline. The results suggested that this strain carries a plasmid (OCT) responsible for oil utilization which is quite stable and might be concerned with antibiotics resistancy.

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Potential of polylactic-co-glycolic acid (PLGA) for delivery Jembrana disease DNA vaccine Model (pEGFP-C1-tat)

  • Unsunnidhal, Lalu;Wasito, Raden;Setyawan, Erif Maha Nugraha;Warsani, Ziana;Kusumawati, Asmarani
    • Journal of Veterinary Science
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    • v.22 no.6
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    • pp.76.1-76.15
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    • 2021
  • Background: The development of a vaccine for Jembrana disease is needed to prevent losses in Indonesia's Bali cattle industry. A DNA vaccine model (pEGFP-C1-tat) that requires a functional delivery system will be developed. Polylactic-co-glycolic acid (PLGA) may have potential as a delivery system for the vaccine model. Objectives: This study aims to evaluate the in vitro potential of PLGA as a delivery system for pEGFP-C1-tat. Methods: Consensus and codon optimization for the tat gene was completed using a bioinformatic method, and the product was inserted into a pEGFP-C1 vector. Cloning of the pEGFP-C1-tat was successfully performed, and polymerase chain reaction (PCR) and restriction analysis confirmed DNA isolation. PLGA-pEGFP-C1-tat solutions were prepared for encapsulated formulation testing, physicochemical characterization, stability testing with DNase I, and cytotoxicity testing. The PLGA-pEGFP-C1-tat solutions were transfected in HeLa cells, and gene expression was observed by fluorescent microscopy and real-time PCR. Results: The successful acquisition of transformant bacteria was confirmed by PCR. The PLGA:DNA:polyvinyl alcohol ratio formulation with optimal encapsulation was 4%:0.5%:2%, physicochemical characterization of PLGA revealed a polydispersity index value of 0.246, a particle size of 925 nm, and a zeta potential value of -2.31 mV. PLGA succeeded in protecting pEGFP-C1-tat from enzymatic degradation, and the percentage viability from the cytotoxicity test of PLGA-pEGFP-C1-tat was 98.03%. The PLGA-pEGFP-C1-tat demonstrated luminescence of the EGFP-tat fusion protein and mRNA transcription was detected. Conclusions: PLGA has good potential as a delivery system for pEGFP-C1-tat.

The Effect of C Language Output Method to the Performance of CGI Gateway in the UNIX Systems (유닉스 시스템에서 C 언어 출력 방법이 CGI 게이트웨이 성능에 미치는 영향)

  • Lee Hyung-Bong;Jeong Yeon-Chul;Kweon Ki-Hyeon
    • The KIPS Transactions:PartC
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    • v.12C no.1 s.97
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    • pp.147-156
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    • 2005
  • CGI is a standard interface rule between web server and gateway devised for the gateway's standard output to replace a static web document in UNIX environment. So, it is common to use standard I/O statements provided by the programming language for the CGI gateway. But the standard I/O mechanism is one of buffer strategies that are designed transparently to operating system and optimized for generic cases. This means that it nay be useful to apply another optimization to the standard I/O environment in CGI gateway. In this paper, we introduced standard output method and file output method as the two output optimization areas for CGI gateways written in C language in the UNIX/LINUX systems, and applied the proposed methods of each area to Debian LINUX, IBM AIX, SUN Solaris, Digital UNIX respectively. Then we analyzed the effect of them focused on execution time. The results were different from operating system to operating system. Compared to normal situation, the best case of standard output area showed about $10{\%}$ improvement and the worst case showed $60{\%}$ degradation in file output area where some performance improvements were expected.

Syntheses and Properties of Copolymers of Tetramethyldisiloxane and 1,4-Bis(dimethylhydroxysilyl)benzene (Tetramethyldisiloxane-1,4-bis(dimethylhydroxysilyl)benzene 공중합체의 합성 및 특성)

  • Kim, Sun-ll;Yun, Young-Jae;Na, Jae-Woon
    • Applied Chemistry for Engineering
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    • v.7 no.3
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    • pp.490-495
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    • 1996
  • Low molecular weight linear chlorine terminated siloxanes (yields;71.2~86.5%) were prepared by reactions of cyclotri-, cyclotetra- and cyclopentasiloxane with dimethyldichlorosilane in the presence of pyridine N-oxide catalyst. The amine terminated siloxane oligomers were obtained in good yields(76.2~85.3%) by the reaction of linear chlorine terminated siloxanes with dimethylamine at $0^{\circ}C$. In this investigation, we have studied on the syntheses and properties of copolymers (yields;58.0~71.0%) obtained from the reaction of amine terminated siloxane oligomers with 1,4-bis(dimethylhydroxysilyl)benzene. The structures and properties of the copolymers were examined by FT-IR, $^1H$-NMR, TGA and DSC. Initial degradation temperatures($T_D{^i}$) of the polymer I and IV were confirmed as 476 and $485^{\circ}C$, respectively. The thermal stabilities of the polymers were found to be increased with increasing n of $(R_2SiO)_n$. The glass transition temperatures(Tg) of the polymers were increased with decreasing n of $(R_2SiO)_n$, and the lowest Tg revealed $-76^{\circ}C$ when n=5.

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Characterization and Production of Thermostable and Acid-stable Extracellular Fibrinolytic Enzymes from Cordyceps militaris

  • Kim, Seon-Ah;Son, Hong-Joo;Kim, Keun-Ki;Park, Hyun-Chul;Lee, Sang-Mong;Cho, Byung-Wook;Kim, Yong-Gyun
    • International Journal of Industrial Entomology and Biomaterials
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    • v.22 no.2
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    • pp.83-93
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    • 2011
  • Biochemical and enzymatic characterization for extracellular protease isolated from Cordyceps militaris cultivated on rice bran medium was investigated. C militaris produced proteolytic enzymes from 10 days after inoculation, maximum enzyme production was found at 25 days. The optimum temperature and pH of proteases production was at $25^{\circ}C$ and pH 7.0, respectively. The protease activity was observed in the four peaks (Pro-I, Pro-II, Pro-III, and Pro-IV) separated through Sephadex G-100 column chromatography. The separated protease was optimally active at $25^{\circ}C$. Optimum pH of the protease was between 7 and 8. Enzyme was also stable over at $30-80^{\circ}C$. The enzyme was highly stable in a pH range of 4-9. Protease activity was found to be slightly decreased by the addition of $Mg^{2+}$, $Mn^{2+}$, $Zn^{2+}$, $Fe^{2+}$ and $Cu^{2+}$, whereas inhibited by the addition of $Ca^{2+}$ and $Co^{2+}$ Protease activity was inhibited by protease inhibitor PMSF. On the other hand, the partially purified protease was investigated on proteolytic protease activity by zymogram gel electrophoresis using three substances (casein, gelatin and fibrin). Four active bands (F-I, FII, F-III, and F-IV) of fibrin degradation were revealed on fibrin zymogram gels. Both of F-II and FIII showed caseinolytic, fibrinolytic and gelatinolytic activities in three gels. Thermostability, pH stability, and pH-thermostability of the enzyme determined the residual fibrinolytic activity also displayed on fibrin zymogram gel. The only one enzyme (F-II) displayed over a broad range of temperature at $30-90^{\circ}C$. The FII displayed fibrinolytic activity in the pH range 3-5, but was inactivated in the range of pH 6-11. The F-I and F-III showed enzyme activity in the pH range of 6-11. In the pH-thermostability, the F-II only kept fibrinolytic activity after heating at $100^{\circ}C$ for 10, 20 and 30 min at pH 3 and pH 7, respectively. On the other hand, the F-II was retained activity until heating for 10 min under pH 11 condition. By using fibrin zymogram gel electrophoresis, extracellular fibrinolytic enzyme F-II from C. militaris showed unusual thermostable under acid and neutral conditions.

Response Time-based Web Service Availability Measurement Method (응답시간기반 웹 서비스 가용성 측정방안)

  • Park, Sang- Kun;Choi, Deok-Jai
    • The KIPS Transactions:PartC
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    • v.10C no.1
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    • pp.61-70
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    • 2003
  • I-Business providers and customers have chosen the service availability as the most important technical parameter which should be included in their SLA to succeed in their business. This means that web Service availability management is crucial to the web-based service providers. Application availability is originally defined as a measure of the fraction of time during a defined period when the service provided is deemed to be better than user expectation of service performance. But, because most web service availability measurement tools simply monitor disconnected state, they do not satisfy user's expectation of extended availability concept. In this paper, We propose the web service availability measurement method which supports extended availability concept. It takes account of performance degradation of web service based on response time, and determines what is the cause of unavailability of the service. We developed a measurement tool, WebSerAvail.

Suppression of Protein Kinase C and Nuclear Oncogene Expression as Possible Action Mechanisms of Cancer Chemoprevention by Curcumin

  • Lin, Jen-Kun
    • Archives of Pharmacal Research
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    • v.27 no.7
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    • pp.683-692
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    • 2004
  • Curcumin (diferuloylmethane) is a major naturally-occurring polyphenol of Curcuma species, which is commonly used as a yellow coloring and flavoring agent in foods. Curcumin has shown anti-carcinogenic activity in animal models. Curcumin possesses anti-inflammatory activity and is a potent inhibitor of reactive oxygen-generating enzymes such as lipoxygenase/cyclooxygenase, xanthine dehydrogenase/oxidase and inducible nitric oxide synthase; and an effective inducer of heme oxygenase-1. Curcumin is also a potent inhibitor of protein kinase C(PKC), EGF(Epidermal growth factor)-receptor tyrosine kinase and LĸB kinase. Subsequently, curcumin inhibits the activation of NF(nucleor factor)KB and the expressions of oncogenes including c-jun, c-fos, c-myc, NIK, MAPKs, ERK, ELK, PI3K, Akt, CDKs and iNOS. It is proposed that curcumin may suppress tumor promotion through blocking signal transduction path-ways in the target cells. The oxidant tumor promoter TPA activates PKC by reacting with zinc thiolates present within the regulatory domain, while the oxidized form of cancer chemopreventive agent such as curcumin can inactivate PKC by oxidizing the vicinal thiols present within the catalytic domain. Recent studies indicated that proteasome-mediated degradation of cell proteins playa pivotal role in the regulation of several basic cellular processes including differentiation, proliferation, cell cycling, and apoptosis. It has been demonstrated that curcumin-induced apoptosis is mediated through the impairment of ubiquitin-proteasome pathway. Curcumin was first biotransformed to dihydrocurcumin and tetrahydrocurcumin and that these compounds subsequently were converted to monoglucuronide conjugates. These results suggest that curcumin-glucuronide, dihydrocurcumin-glucuronide, tetrahydrocurcumin-glucuronide and tetrahydrocurcumin are the major metabolites of curcumin in mice, rats and humans.