• YAKHAK HOEJI
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• v.48 no.1
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• pp.47-54
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• 2004

ClpP is a stress-inducible protein and proteolytic subunit of the ATP-dependent Clp protease in prokaryotes and eukaryotes. Although its physiological roles in bacterial virulence were widely studied in various organsims, including Streptococcus pneumoniae, until now the immunological effect has not been investigated. Here, we have examined the cross reactivity of S. pneumoniae ClpP antibody with other organisms's cell lysate proteins. Although the protein sequence of S. pneumoniae ClpP was highly conserved among various organisms including human, the antibody rasised by S. pneumoniae ClpP was not cross-reacted with other organism's cell lysates, which were Saccharomyces cerevisiae , human lung A549 cell, Bacillus subtilis, Pseuomonas aeruginosa, E. coli, and Salmonella typhi. It was only reacted with S. pneumoniae and Lato-bacillus thermophilus. Thus we examined the immunoprotective effect of ClpP by immunizing mice with the purified ClpP. The mean survival time of mouse was significantly increased with the ClpP immunization. These results suggest that S. pneumoniae ClpP could be used as a vaccine candidate for prevention of S. pneumoniae infection.

• Journal of Life Science
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• v.23 no.8
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• pp.1050-1056
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• 2013

Streptococcus pneumoniae is the leading cause of community-acquired pneumonia throughout the world. The bacteria invade through lung tissue and cause sepsis, shock, and serious sequelae, including rheumatic fever and acute glomerulonephritis. However, the molecular mechanism associated with pneumonia's penetration of lung tissue and invasion of the blood stream are still unclear. We attempted to investigate the host cell response at protein levels to S. pneumoniae D39 invasion using human lung cancer epithelial cells, A549. Streptococcus pneumoniae D39 began to change the morphology of A549 cells to become round with filopodia at 2 hours post-infection. A549 cell proteins obtained at each infection time point were separated by SDS-PAGE and analyzed using MALDI-TOF. We identified several endoplasmic reticulum (ER) resident proteins such as Grp94 and Grp78 and mitochondrial proteins such as ATP synthase and Hsp60 that increased after S. pneumoniae D39 infection. Cytosolic Hsc70 and Hsp90 were, however, identified to decrease. These proteins were also confirmed by Western blot analysis. The identified ER resident proteins were known to be induced during ER stress signaling. These/ data, therefore, suggest that S. pneumoniae D39 infection may induce ER stress.

• Korean Journal of Microbiology
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• v.38 no.1
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• pp.38-44
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• 2002

Pneumococcal surfacce protein A (PspA) is an important virulence factor and an antigenically variable surface protein of the pneumococci. To purify the PspA from S. pneumoniae KNIH1156 , a clinical isolate (type 19F), we have taken advantage of the fact that PspA is released from the surface of pneumococci into the medium by growing in a CDM-ET medium and PspA is capable of binding human lactoferrin, the iron carrier protein. PspA of S. pneumoniae KNIH1156 was purified from culture supernatant by human lactoferrin (hLf) affinity chromatography. The purified PspA was confirmed with anti-PspA antiserum and also had the binding capacity to hLf specifically. To determine whether the purified PspA could elicit protection in mice against pneumococcal inflection, we immunized the mice with purified PspA and subsequently challenged with S. pneumoniae KNIH1156. Immunization with purified PspA protected mice from 500 times the $LD^{50}$ of S. pneumoniae KNIH1156. Therefore, it has been shown that purified PspA fromS. pneumoniae KNIH1156 (type 19F) is a protective immunogen.

• The Journal of Dong Guk Oriental Medicine
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• v.7 no.2
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• pp.121-140
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• 1999

The various oriental herbal medicines, which have usually been used for treatment of reducing fever, purging intense heat and detoxication, were screened to determine the antibacterial activity and the minimal inhibitory concentration against pulmonary disease-causing Klebsiella pneumoniae, Streptococcus pyogenes, and Streptococcus pneumoniae. The results obtained were as follows: 1. Among the 23 oriental medicines tested, the water-soluble extracts of Coptis japonica, Scutellaria baicalensis and Picrorrhiza kurrooa showed the antibacterial activity against K.pneumoniae and that of C. japonica against S. pyogenes. The antibacterial activities of C. japonica, Prunusmume, Schizandra chinesis, Scutellaria baicalensis were also found against S.pneumoniae. When C.japonica was used, the high antibacterial activity was shown against Bacillus subtilis and other extracts showed a little activity against B. subtilis and E. coli as a control. 2. The ethanol-soluble extracts of Patrinia scabriosaefolia, P. mume, S. baicalensis, S. chinesis showed the antibacterial activity against K. pneumoniae and those of S. baicalensis, C. japonica, S. chinesis, P. mume agaist S. pyogenes and S. pnuemoniae. However, those extract showed a little antibacterial activity against B. subtilis and E. coli except for that the extract of C. japonica showed comparatively high growth inhibition of B. subtilis. 3. Among the medicinal herbs tested, the water and ethanol extrats of C. japonica showed very extcellent antibacterial activity against the pathogenic bacteria and controls. 4. When the water-soluble extracts of C. japonica and S. baicalensis, minimal inhibitory concentrations (MICs) against K. pneumoniae were $10mg/m{\ell}$ and $22 mg/m{\ell}$, respectively. The MICs of the ethanol-soluble extracts of P. mume and P. scabriosaefolia were $5mg/m{\ell}$ and $20mg/m{\ell}$, respectively. 5. For the MICs against S.pyogenes, C. japonica showed $15mg/m{\ell}$ with the water-soluble extract and P. mume and C. japonica with the ethanol-souble extract did $5mg/m{\ell}$ and $10mg/m{\ell}$, respectively. 6. For the MICs against S. pneumoniae, C. japonica and P. mume with the water- and ethanol-souble extract showed $5mg/m{\ell}$ and $10mg/m{\ell}$, respectively. As a result, the highest antibacterial activity was found in the water- and ethanol-soluble extracts of C. japonica against pulmonary disease-causing bacteria, K. pneumoniae, S. pyogenes. and S. pnuemoniae. Also, the water- and ethanol-soluble extracts of S.chinesis. P.mume, S.baicalensis, and P.kurrooa showed hight antibacterial activities.

• Microbiology and Biotechnology Letters
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• v.19 no.3
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• pp.259-264
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• 1991

Glutamine synthetase (GS) of Kkbsiellu pmumonzae was purified and identified it's properties. It was determined to be composed of 12 identical subunits and it's molecular weight was about 600,000. It's optimum pH and temperature were identified as pH 7.0 and $37^{\circ}C$ respectively, and also there was no considerable variation of activity between pH 5 and 8. When GS was incubated at $57^{\circ}C$ for 10 min, it's activity was decreased to half of maximum activity. It was observed that K. pneumoniae has adenylylation-deadenylylation system which regulates activity of GS according to the quality and quantity of nitrogen source like GS of E. coli Also it's GS was very similar to that of E. coli. in structure deduced from the immunodiffuslon experiment using anti-E. coli GS antibody.

• Clinical and Experimental Pediatrics
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• v.53 no.2
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• pp.178-183
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• 2010

Purpose : The aim of this study was to identify mutations associated with macrolide resistance in Mycoplasma pneumoniae (MP) and to establish a cultural method to determine antimicrobial susceptibility. Methods : Nasopharyngeal aspirates (NPAs) were collected from 62 children diagnosed with MP pneumonia by a serologic method or polymerase chain reaction. The 23S rRNA and L4 ribosomal protein genes of MP were amplified and sequenced. To identify mutations in these 2 genes, their nucleotide sequences were compared to those of the reference strain M129. MP cultivation was carried out for 32 (28 frozen and 5 refrigerated) NPAs and M129 strain using Chanock's glucose broth and agar plate in a 5% $CO_2$ incubator at $37^{\circ}C$ and examined at 2-3 day intervals for 6 weeks. Results : Among the 62 specimens, 17 had M144V mutations in ribosomal protein L4. The A2064G mutation was observed in 1 specimen; its 23S rRNA gene was successfully sequenced. Culture for MP was successful from the M129 strain and 2 of the 5 NPAs that were refrigerated for no longer than 3 days. However, MP did not grow from the 28 NPAs that were kept frozen at $-80^{\circ}C$ since 2003. Conclusion : We found the M144V mutation of L4 protein to be common and that of domain V of 23S rRNA gene was relatively rare among MP. Studies on the prevalence of macrolide-resistant MP and the relationship between the mutations of 23S rRNA gene and ribosomal protein L4 will aid in understanding the mechanism of macrolide resistance in MP.

• Natural Product Sciences
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• v.13 no.2
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• pp.164-168
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• 2007

To investigate natural antibiotics from plant essential oils and to evaluate their synergism with current antimicrobial drugs in inhibiting antibiotic-resistant strains of Streptococcus pneumoniae. The minimal inhibitory concentrations (MICs) of eleven plant essential oils and their main components were established for two antibiotic-susceptible and two antibiotic-resistant strains of S. pneumoniae, using broth microdilution tests. Potential synergism with oxacillin, norfloxacin, or erythromycin was evaluated using a checkerboard microtitre assay. Among the tested oils, Mentha piperita oil and its main component, menthol, exhibited the strongest inhibitory activities against all of the tested strains. The activity of antibiotics against antibiotic-resistant strains of S. pneumoniae was enhanced significantly by combination with Mentha piperita oils and its main component, menthol. In conclusion, the combination Mentha piperita essential oil or menthol with antibiotics could be used to reduce the effective dose of antibiotic and to modulate the resistance of S. pneumoniae strains.

• YAKHAK HOEJI
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• v.48 no.6
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• pp.345-351
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• 2004

ln the present work to determine effect of a Streptococcus pneumoniae conjugate vaccine, S.pneumoniae capsule attached to the surface protein (JY-Pol) was ex amined. This JY-Pol contained approximately 92% and 6% carbohydrate and protein, respectively. Gel electrophoresis revealed the presence of the surface protein in the JY-Pol. By the double immunodiffusion and isotyping ELISA analyses, administration of JY-Pol that was adsorbed to alum adjuvant (JY-Pol/Alum) into mice induced IgM, IgG, and IgA specific for the S.pneumoniae capsule. The ATCC capsular polysaccharide adsorbed to alum (ATCC-Pol/Alum) provoked only IgM in mice. In survival tests, mice that were immunized with the JY-Pol/Alum before intravenous challenge with live S.pneumoniae survived entire period of 46 day-observation, whereas all mice that received ATCC-Pol/Alum or only diluent instead of the vaccination died within 5 and 12 days, respectively. Results from footpad-edema test showed that JY-Pol/Alum formula provoked the cellular immunity as determined by swelling of the mouse footpad. These data indicate that the naturally conjugated JY- Pol enhances resistance of mice against disseminated pneumococcal disease due to S.pneumoniae by both humoral and cellular immune responses.

• Korean Journal of Clinical Laboratory Science
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• v.40 no.1
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• pp.6-17
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• 2008

From the total 116,429 clinical specimens submitted to "C" hospital from January 2005 to December 2006, 2,195 strains of streptococci were isolated. Twenty four species of Streptococcus were identified with 0.1~19.8% isolation frequencies, of which S. pneumoniae was 19.8%, S. agalactiae 16.2%, S. anginosus 9.8%, S. constellatus 5.0%, S. oralis 3.9%, S. mitis 3.3%, S. pyogenes 2.7%, S. salivarius subsp. salivarius 2.2%, S. sanguinis 1.9%. For S. pneumoniae, clinical specimens showing over 9.0% isolation rate were 82.8% in sputum, 9.2% in blood, and for Streptococcus species other than S. pneumoniae, 18.0% in sputum, 16.0% in urine and 9.7% in blood. The antimicrobial agents that showed over 90.0% susceptibility were cefotaxime, gatilfloxacin, imipenem, levofloxacin, linezolid, moxifloxacin, rifampin and sporfloxacin in S. pneumoniae, ampicillin, cefotaxime, cetriaxone, levofloxacin, linezolid, penicillin, quinupristin/dalfopristin and vancomycin in S. agalactiae, chloramphenicol, clindamycin, levofloxacin and vancomycin in S. anginosus, levofloxacin, vancomycin in S. constellatus subsp. constellatus, vancomycin in S. oralis, vancomycin in S. mitis, chloramphenicol, clindamycin, levofloxacin, quinupristin/dalfopristin and vancomycin in S. pyogenes, chloramphenicol, levofloxacin and vancomycin in S. salivarius subsp. salivarius, chloramphenicol, levofloxacin and vancomycin in S. bovis II, chloramphenicol, levofloxacin, quinupristin/dalfopristin and vancomycin in S. dysgalactiae subsp. dysgalactiae, levofloxacin, chloramphenicol and vancomycin in the whole group of 10 Streptococcus spp. other than Streptococcus pneumoniae.

• YAKHAK HOEJI
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• v.49 no.4
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• pp.312-316
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• 2005

To develop a new natural antibiotics from Koran plant resources for dealing with the current situation regarding the rapid increase in antibiotic-resistant pathogen, the in vitro inhibitory activities of essential oils from the young leaves of Glehnia littoralis (Umbelliferae) as well as its main constituents were evaluated against susceptible and resistant species of Streptococcus pneumoniae. The essential oil fraction of G. littoralis and its main components, $\alpha-and\;\beta-pinene$, exhibited significant inhibitory activities against the antibiotic-susceptible and resistant strains of S. pneumoniae, with MICs (minimum inhibiting concentrations) ranging from 4.0mg/ml to 16mg/ml. No remarkable differences were shown between the susceptible and resistant strains. Moreover, the disk diffusion test disclosed that these inhibitory activities were dose­dependent. Furthermore, data from the checkerboard titer test with FICIs (fractional inhibiting concentration indices) from 0.15 to 0.28 indicated synergisms between norfloxacin and $\alpha-or\;{\beta}-pinene$ in activity against S. pneumoniae KCCM49629 and S. pneumoniae CCARM4059.