• Title/Summary/Keyword: SDS resistance

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Studies on the development of the tick (Haemaphysalis longicornis)-vaccine (I) - Immune responses on the crude soluble - (진드기 백신 개발을 위한 기초연구(I) - 수용성 항원에 대한 면역반응에 관하여 -)

  • Jeong, Woo-seog;Kang, Seung-won;Choi, Eun-jin;Yoon, Yong-dhuk
    • Korean Journal of Veterinary Research
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    • v.36 no.3
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    • pp.693-698
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    • 1996
  • Haemaphysalis longiscornis is the common cattle tick of great economic importance in Korea. Chemical control using dips or sprays has been the traditional method of attempting to kill these ticks during the infestation period. However, the presence of resistant forms to chemical, the rising costs of acaricides and environmental problems have made it almost impossible to use these chemicals on a regular basis according to the pest problem. For this reason, vaccination against ticks and breeding for host resistance against ticks are being studied. In order to determine the common proteins and antigens according to developmental stages, SDS-PAGE and western blotting were performed. In SDS-PAGE 103.3kD and 98.3kD proteins were observed as common proteins, and these proteins were observed as common antigens in western blotting. Unimmunized rabbits were infestated three times with H longicornis. The weight of the second and the third engorged ticks were 0.153g and 0.104g respectively. This weight is 69% and 47% of the first engorged ticks weight respectively. Immunized rabbits by adult ticks antigen and control were infested with H longicornis. The control taked 3-4 days to fully engorge, but the immunized rabbits taked about 7 days. So adult tick antigen may be effective to render the immunity to host.

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Growth hormone therapy in patients with Noonan syndrome

  • Seo, Go Hun;Yoo, Han-Wook
    • Annals of Pediatric Endocrinology and Metabolism
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    • v.23 no.4
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    • pp.176-181
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    • 2018
  • Noonan syndrome (NS) is an autosomal dominant disorder that involves multiple organ systems, with short stature as the most common presentation (>70%). Possible mechanisms of short stature in NS include growth hormone (GH) deficiency, neurosecretory dysfunction, and GH resistance. Accordingly, GH therapy has been carried out for NS patients over the last three decades, and multiple studies have reported acceleration of growth velocity (GV) and increase of height standard deviation score (SDS) in both prepubertal and pubertal NS patients upon GH therapy. One year of GH therapy resulted in almost doubling of GV compared with baseline; afterwards, the increase in GV gradually decreased in the following years, showing that the effect of GH therapy wanes over time. After four years of GH therapy, ~70% of NS patients reached normal height considering their age and sex. Early initiation, long duration of GH therapy, and higher height SDS at the onset of puberty were associated with improved final height, whereas gender, dosage of GH, and the clinical severity did not show significant association with final height. Studies have reported no significant adverse events of GH therapy regarding progression of hypertrophic cardiomyopathy, alteration of metabolism, and tumor development. Therefore, GH therapy is effective for improving height and GV of NS patients; nevertheless, concerns on possible malignancy remains, which necessitates continuous monitoring of NS patients receiving GH therapy.

Isolation of Surfactant-Resistant Pseudomonads from the Estuarine Surface Microlayer

  • Louvado, Antonio;Coelho, Francisco J.R.C.;Domingues, Patricia;Santos, Ana L.;Gomes, Newton C.M.;Almeida, Adelaide;Cunha, Angela
    • Journal of Microbiology and Biotechnology
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    • v.22 no.3
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    • pp.283-291
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    • 2012
  • Bioremediation efforts often rely on the application of surfactants to enhance hydrocarbon bioavailability. However, synthetic surfactants can sometimes be toxic to degrading microorganisms, thus reducing the clearance rate of the pollutant. Therefore, surfactant-resistant bacteria can be an important tool for bioremediation efforts of hydrophobic pollutants, circumventing the toxicity of synthetic surfactants that often delay microbial bioremediation of these contaminants. In this study, we screened a natural surfactant-rich compartment, the estuarine surface microlayer (SML), for cultivable surfactant-resistant bacteria using selective cultures of sodium dodecyl sulfate (SDS) and cetyl trimethylammonium bromide (CTAB). Resistance to surfactants was evaluated by colony counts in solid media amended with critical micelle concentrations (CMC) of either surfactants, in comparison with non-amended controls. Selective cultures for surfactant-resistant bacteria were prepared in mineral medium also containing CMC concentrations of either CTAB or SDS. The surfactantresistant isolates obtained were tested by PCR for the Pseudomonas genus marker gacA gene and for the naphthalene-dioxygenase-encoding gene ndo. Isolates were also screened for biosurfactant production by the atomized oil assay. A high proportion of culturable bacterioneuston was tolerant to CMC concentrations of SDS or CTAB. The gacA-targeted PCR revealed that 64% of the isolates were Pseudomonads. Biosurfactant production in solid medium was detected in 9.4% of tested isolates, all affiliated with genus Pseudomonas. This study shows that the SML is a potential source of surfactant-resistant and biosurfactant-producing bacteria in which Pseudomonads emerge as a relevant group.

STM Studies of Keggin-type and Wells-Dawson-type Heteropolyacid Catalysts (Keggin 형 및 Wells-Dawson 형 헤테로폴리산 촉매의 STM 연구)

  • Park, Gyo Ik;Barteau, Mark A.;Jung, Ji Chul;Song, In Kyu
    • Korean Chemical Engineering Research
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    • v.47 no.2
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    • pp.163-168
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    • 2009
  • Negative differential resistance(NDR) behaviors of Keggin-type and Wells-Dawson-type heteropolyacids with cation, heteroatom, and polyatom substitutions were investigated by scanning tunneling microscopy. A reliable correlation between NDR peak voltage and reduction potential of heteropolyacid catalysts was established. It was found that more reducible heteropolyacid catalyst showed NDR behavior at less negative voltage, regardless of the structural difference. Thus, NDR peak voltage of heteropolyacid catalyst could be utilized as a single correlating parameter for the reduction potential of heteropolyacid catalyst.

Thermostability and Resistance to Proteolysis of L-Asparaginase Purified from Strepfomyces lincolnensis M-20 (Strepfomyces lincolnensis M-20 균주로 부터 분리, 정제된 L-Asparaginase의 열안정성과 단백 가수 분해 효소에 대한 저항성)

  • Kim, Kyoung-Ja
    • YAKHAK HOEJI
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    • v.51 no.3
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    • pp.199-205
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    • 2007
  • Thermostable asparaginase was purified to homogeneity from mesophilic Strepfomyces lincolnensis M-20 by 30${\sim}$70% ammonium sulfate precipitation and asparagine-Sepharose CL 6B affinity column chromatography, The apparent molecular mass of L-asparaginase by SDS-PAGE was found to be 47 kDa, whereas by its mobility on Sephacryl S-300 column was around 180 kDa, indicating that the enzyme at the native stage acts as tetramer, The purified enzyme showed a single band on acrylamide gel electrophoresis. The optimum pH and temperature were pH 9.5 and 55${\circ}$C, respectively. Chemical modification experiments of purified asparagines implied the existence cystein residue located at or near active site. Purified asparaginase retained the 85% of the initial activity after incubation at 90${\circ}$C for 30 min. A correlation between themostability and resistance to proteolysis of commercial asparaginase and purified asparaginase from Strepfomyces lincolnensis M-20 was investigated. Purified thermostable asparaginase was resistant to trypsin and chymotrypsin treatment, while the commercial asparaginase was not themostable and was susceptible to proteolytic treatment with trypsin and chymotrypsin.

Plasmid Profiling and Curing of Lactobacillus Strains Isolated from the Gastrointestinal Tract of Chicken

  • Chin Sieo Chin;Abdullah Norhani;Siang Tan Wen;Wan Ho Yin
    • Journal of Microbiology
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    • v.43 no.3
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    • pp.251-256
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    • 2005
  • In this study, we assessed the susceptibility of 12 Lactobacillus strains, all of which had been isolated from the gastrointestinal tracts of chicken, to three antibiotics (chloramphenicol, erythromycin and tetracycline) used commonly as selective markers in transformation studies of lactic acid bacteria. Among these strains, $17\%,\;58\%,\;and\;25\%$ were found to exhibit a high degree of resistance to $200\;{\mu}g/ml$ of tetracycline, erythromycin, and chloramphenicol, respectively. Seven of the 12 Lactobacillus strains exhibiting resistance to at least $50\;{\mu}g/ml$ of chloramphenicol or erythromycin, and five strains exhibiting resistance to at least $50\;{\mu}g/ml$ of tetracycline, were subsequently subjected to plasmid curing with chemical curing agents, such as novobiocin, acriflavin, SDS, and ethidium bromide. In no cases did the antibiotic resistance of these strains prove to be curable, with the exception of the erythromycin resistance exhibited by five Lactobacillus strains (L. acidophilus I16 and I26, L. fermentum I24 and C17, and L. brevis C10). Analysis of the plasmid profiles of these five cured derivatives revealed that all of the derivatives, except for L. acidophilus I16, possessed profiles similar to those of wild-type strains. The curing of L. acidophilus I16 was accompanied by the loss of 4.4 kb, 6.1 kb, and 11.5 kb plasmids.

Evaluation of friction of ceramic brackets in various bracket-wire combinations (브라켓 각도 변화에 따른 세라믹 브라켓의 마찰력 측정)

  • Cha, Jung-Yul;Kim, Kyung-Suk;Kim, Dong-Choon;Hwang, Chung-Ju
    • The korean journal of orthodontics
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    • v.36 no.2 s.115
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    • pp.125-135
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    • 2006
  • The purpose of this study was to measure and compare the level of frictional resistance generated from three currently used ceramic brackets; 1, Crystaline $V^{(R)}$, Tomy International Inc., Tokyo, Japan; 2, $Clarity^{(R)}$, 3M Unitek, Monrovia, CA, USA; 3, $Inspire^{(R)}$, Ormco, Orange, CA, USA; with composite resin brackets, Spirit, Ormco, Orange, CA, USA; and conventional stainless steel brackets, Kosaka, Tomy International Inc., Tokyo, Japan used as controls. In this experiment, the resistance to sliding was studied as a function of four angulations $(0^{\circ},\;5^{\circ},\;10^{\circ}\;and\;15^{\circ})$ using 2 different orthodontic wire alloys: stainless steel (stainless steel, SDS Ormco, Orange, CA, USA), and beta-titanium (TMA, SDS Ormco, Orange, CA, USA). After mounting the 22 mil brackets to the fixture and $.019{\times}.025$ wires ligated with elastic ligatures, the arch wires were slid through the brackets at 5mm/min in the dry state at $34^{\circ}C$. Silica-insert ceramic brackets generated a significantly lower frictional force than did other ceramic brackets, similar to that of stainless steel brackets. Beta-titanium archwires had higher frictional resistance than did stainless steel, and all the brackets showed higher static and kinetic frictional force as the angulation increased. When the angulation exceeded $5^{\circ}$, the active configuration emerged and frictional force quickly increased by 2.5 to 4.5-fold. The order of frictional force of the different wire-bracket couples transposed as the angle increased. The silica-insert ceramic bracket is a valuable alternative to conventional stainless steel brackets for patients with esthetic demands.

Condition Optimization for Overexpression of the Aklavinone 11-Hydroxylase Gene from Streptomyces peucetius subsp. caesius ATCC 27952 in Escherichia coli. (Streptomyces peucetius subsp. caesius ATCC 27952 유래 Aklavinone 11-Hydroxylase 유전자의 대장균에서의 대량발현과 최적화)

  • 민우근;홍영수;최용경;이정준;홍순광
    • Microbiology and Biotechnology Letters
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    • v.26 no.1
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    • pp.15-22
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    • 1998
  • The dnrF gene, responsible for conversion of aklavinone to $\varepsilon$-rhodomycinone via C-11 hydroxylation, was mapped in the daunorubicin gene cluster of Streptomyces peucetius subsp. caesius ATCC 27952, close to drrAB, one of the anthracycline resistance genes. To characterize the enzymatic properties of the aklavinone 11-hydroxylase, the dnrF gene was overexpressed in Escherchia coli. The pET-22(+) plasmid which has the T7 promoter under the control of lacUV5 gene was used for the overexpression of the dnrF gene, and the recombinant plasmid pET213 that contains the dnrF gene linked to the T7 promoter of pET-22b(+) was introduced into the E. coli BL2l. When the expression of the dnrF gene was induced by IPTG at the final concentration of 1 mM, the induced protein could be detected in SDS-PAGE only in insoluble precipitate. The insoluble protein was electroeluted from the gel and used for the preparation of antiserum in mice. Various culture conditions were tested to maximize the expression of the aklavinone 11-hydroxylase in soluble form. The enzymatic activity was checked by the bioconversion experiment, and the protein was confirmed by the SDS-PAGE and the Western blot analysis. From the analysis of the data, it was concluded that the culture induced with IPTG at the final concentration of 0.02 mM at 37$^{\circ}C$ yielded the best productivity of active form of enzyme.

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Studies on the Antibacterial Activity of Enzymatic Hydrolyzates of Lactoferrin Derived from Bovine Colostrum (유우 Lactoferrin 효소가수분해물 항균 활성에 관한 연구)

  • Han, Su Yeon;Kim, Jong Woo
    • Korean Journal of Agricultural Science
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    • v.25 no.1
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    • pp.52-67
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    • 1998
  • The investigative research on the mammalian milk purely consisted of the physiological quality of lactoferrin was conducted to reveal the antimicrobial ativity of specifically functional foods with antibiotic characteristics as a basic data in food manufacturing. Bovine lactoferrin were isolated from raw milk samples, and was digested with pepsin, trypsin and chymotrypsin. It was necessary then to separate and purify lactoferrin from bovine raw milk, and in order to analyze the antimicrobial activity of the enzyme-treated bovine lactoferrin in their required quantitative fraction. Afterwards Escherichia coli and Staphylococcus aureus was incubated in it. It was that investigated to enzyme-treated fractions molecular weight and the peptide fragment with antimicrobial effect. 1. The purity of enzyme-treated bovine lactoferrin(BLF) was tested by SDS-PAGE. As a results of 12% SDS-PAGE assay, pepsin-treated LF did not exhibited band until if reaches 14 KDa, while trypsin and chymotrypsin treated LF, known to contain the non-digestive lactoferrin exhibited band at a molecular weight of 33 KDa. 2. Bovine lactoferrin was sucessfully purified through the use of Sephadex G-50 Column. In order to assay LF through the Sephadex G-50 column chromatography, the digestive bovine lactoferrin (BLFs) was eluted with a linear gradient of 0.05% Tris-HCl. When the gel-filtration analysis, pepsin, trypsin and chymotrypsin treatments of BLF fragments was showed 2, 3, and 2 peak, respectively. The results of the HPLC analysis confirmed that had a non-digestive lactoferrin receptor, and trypsin and chymotrypsin treated BLFs has an antimicrobial effect. 3. To measure the strength of the antimicrobial effect of enzyme treated lactoferrin it was compared to the antimicrobial activity taking place at the incubated Escherichia coli and Staphylococcus aureus. This might explain the resistance of the microorganisms for peptide fragment. The pepsin-treated of bovine lactoferrin was markedly reduced by incubation of the cells. Trypsin-treated of BLF was similar to chymotrypsin-treated of BLF. However, trypsin and chymotrypsin treatments of BLFs were showed the antimicrobial effect until eight hours incubation for native bovine lactoferrin. Therefore the enzyme-treated lactoferrin have an antimicrobial effect even non-digestive lactoferrin. 4. The digestive bovine lactoferrin fragments assay was carried out by the use of Sephadex G-50 column chromatography and SDS-PAGE. The pepsin and chymotrypsin-treated fragments has a low molecular weight and trypsin-treated lactoferrin was only showed a band. It was described that characteristics of digestive protein. It appeared that there may be a relation between virulence and resistance to enzyme-treated BLF.

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Cloning of tlrD, 23S rRNA Monomethyltransferase Gene, Overexpression in Eschepichia coli and Its Activity (235 rRNA Monomethyltransferase인 tlrD의 클로닝, 이의 대장균에서 대량생산과 활성 검색)

  • Jin, Hyung-Jong
    • Korean Journal of Microbiology
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    • v.43 no.3
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    • pp.166-172
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    • 2007
  • ERM proteins transfer the methyl group to $A_{2058}$ in 23S rRNA, which reduces the affinity of MLS (macrolide-lincosamide-streptogramin B) antibiotics to 23S rRNA, thereby confer the antibiotic resistance on micro-organisms ranging from antibiotic producers to pathogens and are classified into monomethyltransferase and dimethyltransferase. To investigate the differences between mono- and dimethyltransferase, tirD, a representative monomethylase gene was cloned in Escherichia coli from Streptomyces fradiae which contains ermSF, dimethylase gene as well to overexpress the TlrD for the first time. T7 promoter driven expression system successfully overexpress tlrD as a insoluble aggregate at $37^{\circ}C$ accumulating to around 55% of the total cell protein but unlike ErmSF, culturing at temperature as low as $18^{\circ}C$ did not make insoluble aggregate of protein into soluble protein. Coexpression of Thioredoxin and GroESL, chaperone was not helpful in turning into soluble protein either as in case of ErmSF. These results might suggest that differences between mono- and dimethylase could be investigated on the basis of the characteristics of protein structure. However, a very small amount of soluble protein which could not be detected by SDS-PAGE conferred antibiotic resistance on E. coli as in ErmSF which was expected from the activity exerted by monmethylase in a cell.