• Molecules and Cells
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• v.23 no.2
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• pp.239-245
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• 2007

DnaK is a major antigen in Streptococcus pneumoniae, and is induced by a minor shift in temperature (30 to $37^{\circ}C$) but not by ethanol shock. Although HrcA in the presence of $Ca^{{+}{+}}$ represses the expression of both groEL and hrcA, the control of transcription of the dnaK operon is not completely understood. In this study, the dnaK operon of S. pneumoniae (5' hrcA-grpE-dnaK-dnaJ) was cloned and analyzed. It contains large intergenic regions in grpE/dnaK and dnaK/dnaJ. Pulse labeling with [$^{35}S$]-methionine and immunoblot analyses revealed the presence of higher levels of DnaK than of HrcA even in the presence of $Ca^{{+}{+}}$ after heat shock suggesting that $Ca^{{+}{+}}$ differentially regulates the heat shock responses of hrcA and dnaK. By blocking de novo mRNA synthesis with rifampin it was shown that neither the hrcA nor the groEL transcripts were stabilized by heat shock even though dnaK transcripts were stabilized. We conclude that S. pneumoniae uses fine regulation of the transcription of the individual genes of the tetracistronic dnaK operon to cope with the various stresses experienced during infections.

• YAKHAK HOEJI
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• v.49 no.6
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• pp.484-489
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• 2005

Streptococcus pmeumoniae is the leading cause of pneumonia and bacterial meningitis. The current polysaccharide vaccine has been reported ineffective in elderly adults and children less than 2 years of age. Thus, in recent many researchers have been focused on a different approach, DNA vaccine. In our laboratory we developed a Streptococcus pneumoniae DNA (SPDNA) vaccine. This SPDNA vaccine was formulated by inserting the region encoding part of the capsule in the S. pneumoniae into the LAMP-1. In present work, with use of the SPDNA vaccine we attempted to establish a certain methodology useful for evaluation of effectiveness and immunoresponse of a DNA vaccine. Results showed that the subcutaneous route was the most effective for production of antisera specific for S. pneumoniae in mice. By isotyping analyses, IgM, IgGl, IgG2a, and IgG2b were determined. In addition, INF-$\gamma$ and IL-4 were predominantly detected. Combination of those data resulted in a pattern of IgGl < IgG2a=IgG2b and INF$\gamma\>$ >IL-4, which indicates the inmmunity towards the Thl response predominantly; furthermore, the SPDNA vaccination induced resistance of the CD4+T lymphocyte-depleted mice against disseminated pneumococcal infection. These data appear to be possibly due to activation of CDS8+T cell-activation. Taken together, this methodology can be applied for evaluating efficacy and mode of action of a DNA vaccine as minimum critera.

• Proceedings of the PSK Conference
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• 2003.10b
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• pp.70.2-71
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• 2003

Quinolone resistance in Streptococcus pneumoniae is related to mutations in the DNA gyrase and topoisomerase IV genes. DW-286a displayed potent activity against S. pneumoniae C9211 (MIC, 0.015 ${\mu}$g/ml) compared with gemifloxacin (MIC, 0.06 ${\mu}$g/ml). This study was performed to analyze the ability of DW-286a to cause resistance development in S. pneumoniae and to establish whether DNA gyrase or topoisomerase IV is primary target. DW-286a resistant mutants of S. pneumoniae C9211 were generated by stepwise selection at increasing drug concentration. (omitted)

• Journal of Life Science
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• v.28 no.1
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• pp.99-104
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• 2018

Streptococcus pneumoniae is one of the major pathogens in community-acquired diseases, and it contains several factors that promote its pathogenesis, including pneumolysin (PLY). PLY is a member of the cholesterol-dependent cytolysin family, which attacks cholesterol-containing membranes, thereby forming ring-shaped pores. Thus, it is a major key target for vaccines against pneumococcal disease. We cloned the PLY gene from S. pneumoniae D39 and inserted it into the pQE-30 vector. Recombinant PLY (rPLY) was overexpressed in Escherichia coli M15 and purified by $Ni^{2+}$ affinity chromatography. Similarly, a PLY-EGFP fusion gene was produced by inserting the EGFP gene at the 3' end of the PLY gene in the same vector, and the recombinant protein was purified. Sodium dodecyl sulfate - polyacrylamide gel electrophoresis (SDS-PAGE) showed that both recombinant proteins were purified. rPLY exhibited significant hemolytic activity against 1% human red blood cells (RBCs). Complete hemolysis was obtained at 500 ng/ml, and 50% hemolysis was found with a 240 ng/ml concentration. In contrast, rPLY-EGFP did not show hemolytic activity. However, rPLY-EGFP did bind the RBC membrane, indicating that rPLY-EGFP lost hemolytic activity via EGFP fusion, while retaining its membrane-binding ability. These data suggest that PLY's C terminus is important for its hemolytic activity. Therefore, these two recombinant proteins can be extremely useful for investigating the toxin mechanism of PLY and cell damage during pneumonia.

• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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• v.12 no.1
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• pp.18-35
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• 1999

Coptidis Rhizoma, Fraxini Cortex, Jinpisan(秦皮散) have been as eye washes of inflammatory eye disease in the oriental medicine. Especially Jinpisan(秦皮散) has been used for the disease which is similar to Peudomonas aeruginisa keratitis. This research was attempted to investigate the effect of Coptidis Rhizoma, Fraxini Cortex, Jinpisan(秦皮散), on Peudoronas aeruginisa keratitis. Pseudomonas aeruginosa keratitis causes a deep rapid intense ulceration which often leads to perforation of the cornea within 48 hours. In this research, we induced keratits in the rabbits by inoculating Pesudomonas aeruginosa(9027) and observed the effect on the keratitis and the irritation against the external eye. Also we mesured the minimum inhibitory consentration(MIC) of Coptidis Rhizoma, Fraxini Cortex, Jinpisan(秦皮散) by agar diliution method and the anti-bacterial activites by disk method. The tested bacteria were as follows : a) Pseudomonas aeruginosa (9027), b) Streptococcus pneumoniae(6303), c) Staphylococcus epidermidis(12228), d) Staphylococcus aureus(6538P). The results were as follows ; 1. The groups which were applied eye washes of Fraxini Cortex, Jinpisan reavealed a significant effect, but the group applied eye wash of Coptidis Rhizoma reveaded no effect on Pseudomonas aeruginosa keratitis. 2. Applying eye washes of Coptidis Rhizoma, Fraxini Cortex, Jinisan revealed an irritation against external eyes. 3. Coptidis Rhizoma showed an anti-bacterial activity on Pseudomonas aeruginosa, Streptococcus pneumoniae, Staphylococcus epidermidis, Staphylucoccus aureus by agar diliution method 4. Coptidis Rhizoma showed an anti-bacterial activity on Pseudomonas aeruginosa, Streptococcus pneumoniae, Staphylococcus epidermidis, Staphylococcus aureus by disk method. 5. Fraxini Cortex showed an anti-bacterial activity on Pseudomonas aeruginosa, Streptococcus pneumoniae, Staphylococcus epidermidis, Staphylococcus aureus by agar diliution method 6. Fraxini Cortex showed an anti-bacterial activity on Pseudomonas aeruginosa, Sireptococcus pneumoniae, Staphylococcus epidermidis, Staphy1ococcus aureus by disk method. 7. Jinpisan showed an anti-bacterial activity on Pseudomonas aeruginosa, Streptococcus pneumoniae, Staphylococcus epidermidis, Staphylococcus aureus by agar diliution method. 8. Jinpisan showed an anti-bacterial activity on Pseudomonas aeruginosa, Streptococcus pneumoniae, Staphylococcus epidermidis, Staphylococcus aureus by disk method. According to the above results, Fraxini Cortex, Jinpisan(秦皮散) are recognized to have an effective treatment on the Pesudomonas aeruginosa keratitis, so this experiment is thought to be a basic ingredient in proving the effect of Fraxini Cortex, Jinpisan which is applied many in documents and clinical medicine. In the comparison of anti-bacterial activity and results of treatment on the Pesudomonas aeruginosa keratitis, Jinpisan(秦皮散) was more effective than Coptidis Rhizoma, Fraxini Cortex.

• YAKHAK HOEJI
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• v.54 no.2
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• pp.122-125
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• 2010

The essential oil fraction was extracted from the aerial parts of the plant by steam distillation method and its composition was analyzed by GC-MS (gas chromatography-mass spectrometry) which led to the identification of 43 compounds. Dehydroelsholtzia ketone (56.81%) and elsholtzia ketone (30.05%) were identified as the predominant components of this oil. The antibacterial activities of the essential oil fraction were assessed by micro-dilution tests against antibioticsusceptible and -resistant strains of Streptococcus pneumoniae, Staphylococcus aureus, Salmonella enteritidis, and S. typhimurium. The oil inhibited most of the tested strains significantly resulting MICs (minimum inhibiting concentrations) between 2 mg/ml and >16 mg/ml. In most cases of this study Streptococcus pneumoniae and Staphylococcus aureus showed higher sensitivity to this oil than Salmonella 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.

• Journal of Microbiology
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• v.44 no.4
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• pp.375-382
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• 2006

From its initial colonization to causation of disease, Streptococcus pneumoniae has evolved strategies to cope with a number of stressful in vivo environmental conditions. In order to analyze a global view of this organism's response to heat shock, we established a 2-D electrophoresis proteome map of the S. pneumoniae D39 soluble proteins under in vitro culture conditions and performed the comparative proteome analysis to a 37 to $42^{\circ}C$ temperature up-shift in S. pneumoniae. When the temperature of an exponentially growing S. pneumoniae D39 culture was raised to $42^{\circ}C$, the expression level of 25 proteins showed changes when compared to the control. Among these 25 proteins, 12 were identified by MALDI-TOF and LC-coupled ESI MS/MS. The identified proteins were shown to be involved in the general stress response, energy metabolism, nucleotide biosynthesis pathways, and purine metabolism. These results provide clues for understanding the mechanism of adaptation to heat shock by S. pneumoniae and may facilitate the assessment of a possible role for these proteins in the physiology and pathogenesis of this pathogen.

• Korean Journal of Microbiology
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• v.27 no.2
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• pp.162-167
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• 1989

Western blot analysis of lysates of Streptococcus pneumoniae revealed a single polypeptide species that cross-reacted with E. coli RecA antiserum. The apparent molecular weight of this putative RecA protein analog (RecAsp) was 24, 000 smaller than any other known RecA analogue. The RecAsp protein was present at the same level in competent and non-competent cells.

• Interdisciplinary Bio Central
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• v.4 no.4
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• pp.11.1-11.8
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• 2012

Genes that are indispensable for survival are referred to as essential gene. Due to the momentous significance of these genes for cellular activity they can be selected potentially as drug targets. Here in this study, an essential gene for Streptococcus suis was predicted using coherent statistical analysis and powerful genome comparison computational method. At first the whole genome protein scatter plot was generated and subsequently, on the basis of statistical significance, a reference genome was chosen. The parameters set forth for selecting the reference genome was that the genome of the query (Streptococcus suis) and subject must fall in the same genus and yet they must vary to a good degree. Streptococcus pneumoniae was found to be suitable as the reference genome. A whole genome comparison was performed for the reference (Streptococcus pneumoniae) and the query genome (Streptococcus suis) and 14 conserved proteins from them were subjected to a screen for potential essential gene property. Among those 14 only one essential gene was found to be with impressive similarity score between reference and query. The essential gene encodes for a type of 'Clp protease'. Clp proteases play major roles in degrading misfolded proteins. Results found here should help formulating a drug against Strptococcus suis which is responsible for mild to severe clinical conditions in human. However, like many other computational studies, the study has to be validated furthermore through in vitro assays for concrete proof.