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Alteration of The Quaternary Structure of Human UDP-Glucose Dehydrogenase by a Double Mutation

  • Huh, Jae-Wan;Yang, Seung-Ju;Hwang, Eun-Young;Choi, Myung-Min;Lee, Hyun-Ju;Kim, Eun-A;Choi, Soo-Young;Choi, Jene;Hong, Hea-Nam;Cho, Sung-Woo
    • BMB Reports
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    • v.40 no.5
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    • pp.690-696
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    • 2007
  • There are conflicting views for the polymerization process of human UDP-glucose dehydrogenase (UGDH) and no clear evidence has been reported yet. Based on crystal coordinates for Streptococcus pyogenes UGDH, we made double mutant A222Q/S233G. The double mutagenesis had no effects on expression, stability, and secondary structure. Interestingly, A222Q/S233G was a dimeric form and showed an UGDH activity, although it showed increased $K_m$ values for substrates. These results suggest that Ala222 and Ser233 play an important role in maintaining the hexameric structure and the reduced binding affinities for substrates are attributable to its altered subunit communication although quaternary structure may not be critical for catalysis.

Genetic characterization of methicillin-resistant Staphylococcus aureus from humans and animals within the community (지역사회 내 일반인 및 동물에서 methicillin 내성 Staphylococcus aureus의 유전학적 특성)

  • Cho, Jae-Keun;Kim, Jin-Hyun;Sung, Myung-Suk;Kim, Ki-Seuk
    • Korean Journal of Veterinary Research
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    • v.51 no.4
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    • pp.267-275
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    • 2011
  • Methicillin-resistant Staphylococcus (S.) aureus (MRSA) is one of the most important nosocomial pathogens worldwide and the emergence of this strain has become a major clinical problem. In this study, we investigated the prevalence of MRSA and their genetic characteristics in 69 S. aureus isolated from humans and animals. In human isolates, higher antimicrobial resistance rates were observed against penicillin (80.6%), followed by erythromycin (11.9%) and tetracycline (9.0%). All of them were susceptible to clindamycin, enrofloxacin, novobiocin, pirlimycin, trimethoprim/sulfamethoxazole and vancomycin. The resistance patterns in animal isolates were similar to those of human isolates. Two (2.9%) MRSA strains were isolated from human (n = 1) and animal (n = 1), and these isolates were confirmed as carrying the mecA gene. One isolate originating from human was resistant to 7 drugs and the other isolate derived from animal was resistant to 11 drugs. Staphylococcal cassette chromosome mec (SCCmec) variant IIIB was identified in animal isolate but SCCmec type of an isolate from human was not exactly determined. Two MRSA isolates showed unrelated PFGE pattern between them. Our results indicated although the frequency of MRSA isolates from humans and animals was low, a continuous surveillance and monitoring should be called for to prevent the contamination and spread of MRSA in the community. To our knowledge, this is the first time that SCCmec type variant IIIB was detected from animals in Korea.

A Study of the Changes of Game Music (게임음악의 변천에 대한 고찰)

  • Lee, Jeong-Hyeok
    • Journal of Korea Game Society
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    • v.12 no.1
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    • pp.103-111
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    • 2012
  • Games in 1970s have used the savings of analog wavelength as in music and the like on hardwares of small cassette, gramophone, and the like. Due to these configuration elements, durability has to be declined so much. In the event of using the music on the video game, more affordable method is to use the computer chip to convert the analog sound into the computer code to convert into the electric wavelength to send to the speaker. The sound effect of the game is generated in this method. The technical limit has been gradually overcome to grant more freedom to the composers and the sound track pre-recorded on the optic disc and the like has emerged. The game developers of today have made several attempts on the technology to produce the game music. This study has contemplated the process of advancement in the change of game music production with the influence on technology and business.

Characterization and Resistance Mechanisms of A 5-fluorouracil-resistant Hepatocellular Carcinoma Cell Line

  • Gu, Wei;Fang, Fan-Fu;Li, Bai;Cheng, Bin-Bin;Ling, Chang-Quan
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.9
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    • pp.4807-4814
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    • 2012
  • Purpose: The chemoresistance of human hepatocellular carcinoma (HCC) to cytotoxic drugs, especially intrinsic or acquired multidrug resistance (MDR), still remains a major challenge in the management of HCC. In the present study, possible mechanisms involved in MDR of HCC were identified using a 5-fluorouracil (5-FU)-resistant human HCC cell line. Methods: BEL-7402/5-FU cells were established through continuous culturing parental BEL-7402 cells, imitating the pattern of chemotherapy clinically. Growth curves and chemosensitivity to cytotoxic drugs were determined by MTT assay. Doubling times, colony formation and adherence rates were calculated after cell counting. Morphological alteration, karyotype morphology, and untrastructure were assessed under optical and electron microscopes. The distribution in the cell cycle and drug efflux pump activity were measured by flow cytometry. Furthermore, expression of potential genes involved in MDR of BEL-7402/5-FU cells were detected by immunocytochemistry. Results: Compared to its parental cells, BEL-7402/5-FU cells had a prolonged doubling time, a lower mitotic index, colony efficiency and adhesive ability, and a decreased drug efflux pump activity. The resistant cells tended to grow in clusters and apparent changes of ultrastructures occurred. BEL-7402/5-FU cells presented with an increased proportion in S and G2/M phases with a concomitant decrease in G0/G1 phase. The MDR phenotype of BEL-7402/5-FU might be partly attributed to increased drug efflux pump activity via multidrug resistance protein 1 (MRP1), overexpression of thymidylate synthase (TS), resistance to apoptosis by augmentation of the Bcl-xl/Bax ratio, and intracellular adhesion medicated by E-cadherin (E-cad). P-glycoprotein (P-gp) might play a limited role in the MDR of BEL-7402/5-FU. Conclusion: Increased activity or expression of MRP1, Bcl-xl, TS, and E-cad appear to be involved in the MDR mechanism of BEL-7402/5-FU.

Efficient transgene expression system using a cumate-inducible promoter and Cre-loxP recombination in avian cells

  • Park, Tae Sub;Kim, Si Won;Lee, Jeong Hyo
    • Asian-Australasian Journal of Animal Sciences
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    • v.30 no.6
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    • pp.886-892
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    • 2017
  • Objective: Transgenic technology is widely used for industrial applications and basic research. Systems that allow for genetic modification play a crucial role in biotechnology for a number of purposes, including the functional analysis of specific genes and the production of exogenous proteins. In this study, we examined and verified the cumate-inducible transgene expression system in chicken DF1 and quail QM7 cells, as well as loxP element-mediated transgene recombination using Cre recombinase in DF1 cells. Methods: After stable transfer of the transgene with piggyBac transposon and transposase, transgene expression was induced by an appropriate concentration of cumate. Additionally, we showed that the transgene can be replaced with additional transgenes by co-transfection with the Cre recombinase expression vector. Results: In the cumate-GFP DF1 and QM7 cells, green fluorescent protein (GFP) expression was repressed in the off state in the absence of cumate, and the GFP transgene expression was successfully induced in the presence of cumate. In the cumate-MyoD DF1 cells, MyoD transgene expression was induced by cumate, and the genes controlled by MyoD were upregulated according to the number of days in culture. Additionally, for the translocation experiments, a stable enhanced green fluorescent protein (eGFP)-expressing DF1 cell line transfected with the loxP66-eGFP-loxP71 vector was established, and DsRed-positive and eGFP-negative cells were observed after 14 days of co-transfection with the DsRed transgene and Cre recombinase indicating that the eGFP transgene was excised, and the DsRed transgene was replaced by Cre recombination. Conclusion: Transgene induction or replacement cassette systems in avian cells can be applied in functional genomics studies of specific genes and adapted further for efficient generation of transgenic poultry to modulate target gene expression.

The Effects of Music Therapy on Cognitive Function and Depression in Demented Old Adults (음악요법이 치매노인의 인지기능과 우울에 미치는 효과)

  • Gwon, Ja-Youn;Kim, Jung-Soo
    • Research in Community and Public Health Nursing
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    • v.9 no.2
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    • pp.336-349
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    • 1998
  • The purpose of this study was to test the effects of music therapy on cognitive function and depression in demented old adults. This study was made with one -group in a pre- and post-test design. The subjects were seven demented old adults over, sixty-five years and with mild to moderate cognitive impairment, residing at a nursing home. Music therapy was given by one researcher and one research assistant for thirty to forty minutes twice a week for 4 months. Music therapy was conducted with the subjects both listening and singing with a cassette player and a double-handed drum. In order to evaluate the effects of music, we measured the level of cognitive function and depression at the beginning and at the end of the music therapy session by means of an MMSE- K developed by Kwon and Park and the Depression Inventory developed by Chon. The Data were analyzed using descriptive statistics and a paired t - test analysis using a SPSS PC package. The results are as follows: 1) The subjects of the music therapy showed improvement in cognitive function. The MMSE-K score was significantly increased after music therapy. Especially, memory recall was very significantly. 2) The subjects of the music therapy showed a slight decrease in depression. However, there was no significant difference in the degree of depression between mean scores measured before and after music therapy. The results suggest that music therapy is effective in improving and maintaining cognitive function in demented old adults. And we suggest that long-term music therapy will be required to improve depression in demented old adults. These findings are encouraging the idea that music therapy may improve cognitive impairment.

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Screening of Bacterial Surface Display Anchoring Motif Using Tetrameric β-galactosidase in Bacillus subtilis Spore (Tetrameric β를 이용한 고초균 포자에서의 미생물 표면 발현 모체 선별)

  • Kim, June-Hyung;Pan, Jae-Gu;Kim, Byung-Gee
    • KSBB Journal
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    • v.26 no.3
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    • pp.199-205
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    • 2011
  • Using tetrameric ${\beta}$-galactosidase as a model protein, anchoring motives were screened in Bacillus subtilis spore display system. Eleven spore coat proteins were selected considering their expression levels and the location in the spore coat layer. After chromosomal single-copy homologous integration in the amyE site of Bacillus subtilis chromosome, cotE and cotG were chosen as possible spore surface anchoring motives with their higher whole cell ${\beta}$-galactosidase activity. PAGE and Wester blot of extracted fraction of outer layer of purified spore, which express CotE-LacZ or CotG-LacZ fusion verified the existence of exact size of fusion protein and its location in outer coat layer of purified spore. ${\beta}$-galactosidase activity of spore with CotE-LacZ or CotG-LacZ fusion reached its highest value around 16~20 h of culture time in terms of whole cell and purified spore. After intensive spore purification with lysozyme treatment and renografin treatment, spore of BJH135, which expresses CotE-LacZ, retained only 1~2% of its whole cell ${\beta}$-galactosidase activity. Whereas spore of BJH136, which has cotG-lacZ cassette in the chromosome, retained 10~15% of its whole cell ${\beta}$-galactosidase activity, proving minor perturbation of CotG-LacZ, when incorporated in the spore coat layer of Bacillus subtilis compared to CotE-LacZ. Usage of Bacillus subtilis WB700, of which 7 proteases are knocked-out and thereby resulting in 99.7% decrease in protease activity of the host, did not prevent the proteolytic degradation of spore surface expressed CotG-LacZ fusion protein.

Effect of Listening to Music on Speech Anxiety among Middle-school Female Students (음악청취가 중학생의 발표불안에 미치는 영향)

  • Oh Yoo-Suk;Sohn Jin-Hun;Jang Eun-Hye;Suk Ji-A;Lee Ok-Hyun
    • Science of Emotion and Sensibility
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    • v.7 no.4
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    • pp.43-49
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    • 2004
  • This study investigated the effect of listening to music on the reduction of public speech anxiety among middle-school female students. The subjects were 66 female first graders, 33 of the experimental and another 33 of the control group selected from two classes of C middle school in Chonan city. The level of anxiety (SAS) was rated by the subjects through self-report and the speech behavior (SBES) of the subjects were evaluated by a teacher before, a week after and two weeks after the speech trials. 64 pieces of music were selected based on the music therapy-related references. After 23 out of 64 pieces were selected for the preliminary experiment and 7 pieces as having positive effects through evaluating those 23 pieces in the other class in the same gaders were finally selected. Subjects listened to music for 40 min two times a day for two weeks with a cassette player in the classroom. The result yielded the followings: 1) Self-reported public speech anxiety decreased both in the experimental and control groups. However, there found more decrease in the music-listening group than in the non-listening music group. 2) The public speech behavior improved both in the experimental and control groups with on difference between the two groups. This suggests a possibility that SBES may not be a accurate measure to evaluate the speech anxiety. We conclude that two-week listening to music has the effect reduction on speech anxiety.

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Investigation of Herb-Drug Interactions between Korean Red Ginseng Extract and five CYP Substrates by LC-MS/MS

  • Jo, Jung Jae;Lee, Sangkyu
    • Mass Spectrometry Letters
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    • v.8 no.4
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    • pp.98-104
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    • 2017
  • Ginseng (Panax ginseng Meyer) is a well-known health functional food used as a traditional herbal drug in Asian countries owing to its diverse pharmacological effects. Herb-drug interactions may cause unexpected side effects of co-administered drugs by the alteration of pharmacokinetics through effects on cytochrome P450 activity. In this study, we investigated the herb-drug interactions between Korean red ginseng extract (KRG) and five CYP-specific probes in mice. The pharmacokinetics of KRG extract induced-drug interactions were studied by cassette dosing of five CYP substrates for CYP1A, 2B, 2C, 2D, and 3A and the LC-MS/MS analysis of the blood concentration of metabolites of each of the five probes. The linearity, precision, and accuracy of the quantification method of the five metabolites were successfully confirmed. The plasma concentrations of five metabolites after co-administration of different doses of the KRG extract (0, 0.5, 1, and 2 g/kg) were quantified by LC-MS/MS and dose-dependent pharmacokinetic parameters were determined. The pharmacokinetic parameters of the five metabolites were not significantly altered by the dose of the KRG extract. In conclusion, the single co-administration of KRG extract up to 2 g/kg in vivo did not cause any significant herb-drug interactions linked to the modulation of CYP activity.

Isolation and Characterization of the Colletotrichum acutatum ABC Transporter CaABC1

  • Kim, Suyoung;Park, Sook-Young;Kim, Hyejeong;Kim, Dongyoung;Lee, Seon-Woo;Kim, Heung Tae;Lee, Jong-Hwan;Choi, Woobong
    • The Plant Pathology Journal
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    • v.30 no.4
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    • pp.375-383
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    • 2014
  • Fungi tolerate exposure to various abiotic stresses, including cytotoxic compounds and fungicides, via their ATP-driven efflux pumps belonging to ATP-binding cassette (ABC) transporters. To clarify the molecular basis of interaction between the fungus and various abiotic stresses including fungicides, we constructed a cDNA library from germinated conidia of Colletotrichum acutatum, a major anthracnose pathogen of pepper (Capsicum annum L.). Over 1,000 cDNA clones were sequenced, of which single clone exhibited significant nucleotide sequence homology to ABC transporter genes. We isolated three fosmid clones containing the C. acutatum ABC1 (CaABC1) gene in full-length from genomic DNA library screening. The CaABC1 gene consists of 4,059 bp transcript, predicting a 1,353-aa protein. The gene contains the typical ABC signature and Walker A and B motifs. The 5'-flanking region contains a CAAT motif, a TATA box, and a Kozak region. Phylogenetic and structural analysis suggested that the CaABC1 is a typical ABC transporter gene highly conserved in various fungal species, as well as in Chromista, Metazoans, and Viridiplantae. We also found that CaABC1 was up-regulated during conidiation and a minimal medium condition. Moreover, CaABC1 was induced in iprobenfos, kresoxim-methyl, thiophanate-methyl, and hygromycin B. These results demonstrate that CaABC1 is necessary for conidiation, abiotic stress, and various fungicide resistances. These results will provide the basis for further study on the function of ABC transporter genes in C. acutatum.