• Pediatric Infection and Vaccine
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• v.26 no.3
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• pp.188-193
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• 2019

Since the implementation of conjugate Haemophilus influenzae serotype b (Hib) vaccine, the rate of infections caused by Hib has dramatically decreased, and the proportion of infections caused by non-type b H. influenzae has increased. Cerebral venous sinus thrombosis (CVST) is rare; however, it should be considered as a potential complication of bacterial meningitis. Herein, we report about a child who developed CVST after being diagnosed with H. influenzae serotype f meningitis.

• Journal of Microbiology and Biotechnology
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• v.14 no.4
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• pp.836-843
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• 2004

Nontypeable H. influenzae (NTHi), a Gram-negative obligate human pathogen, causes pneumonia, chronic bronchitis, and otitis media, and the respiratory epithelium is the first line of defense that copes with the pathogen. In an effort to identify transcriptional responses of human respiratory epithelial cells to infection with NTHi, we examined its differential gene expression using high density cDNA microarrays. BEAS-2B human bronchial epithelial cells were exposed to NTHi for 3 hand 24 h, and the alteration of mRNA expression was analyzed using microarrays consisting of 8,170 human cDNA clones. The results indicated that approximately 2.6% of the genes present on the microarrays increased in expression over 2-fold and 3.8% of the genes decreased during the 24-h infection period. Upregulated genes included cytokines (granulocyte-macrophage colony stimulating factor 2, granulocyte chemotactic protein 2, IL-6, IL-10, IL-8), transcription factors (Kruppel-like factor 7, CCAAT/enhancer binding protein $\beta$, E2F-1, NF-$\kappa$B, cell surface molecules (CD74, ICAM-1, ICAM-2, HLA class I), as well as those involved in signal transduction and cellular transport. Selected genes were further confirmed by reverse-transcription-PCR. These data expand our knowledge of host cellular responses during NTHi infection and should provide a molecular basis for the study of host-NTHi interaction.

• Korean Journal of Microbiology
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• v.39 no.1
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• pp.36-39
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• 2003

The rapid and reliable 16S rDNA multiplex polymerase chain reaction (PCR) assay was established to characterize bacterial etiologies of middle ear effusion. These etiologies included Haemophilus influenzae, Moraxella catarrhalis and Streptococcus pneumonia, which were detected in middle-ear effusion (MEE) samples taken from patient with otitis media. A total of 39 MEE samples were aspirated from 26 patients. DNA was extracted from MEE samples, and PCR was done with DNA extracts by using the common primers, which is localized at C4 region in the 16S rDNA gene of all bacterial species, and species-specific primers: (i) Haemophilus-specific primer, (ii) Moraxella- specific primer, and (iii) Streptococcus-specific primer. Among 39 samples tested, 24 (61.5%) were positive for H. influenzae, 10 (25.6%) were positive for M. catarrhalis, 3(7.7%) were positive for S. pneumonia, and 11 (28%) were negative for 165 rDNA multiplex PCR reaction. Nine samples (28.6%) exhibited a mixed infection and were positive for both H. infuenzae and M. catarrhalis. We suggested that 16S rDNA multiplex PCR is a useful method to identify rapidly for rapid identification of the pathogenic bacteria and characterization of bacterial etiologies of middle ear effusion.

• Clinical and Experimental Pediatrics
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• v.51 no.10
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• pp.1077-1084
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• 2008

Purpose : Despite the seriousness of bacterial meningitis in children, there is little information on the incidence, causative organisms, mortality rate and age distribution. We studied the frequency by age group and causal pathogens, and clinical characteristics in children with bacterial meningitis in the private sector in Korea. Methods : The medical records containing the data on bacterial meningitis patients under 18 years of age confirmed by cerebrospinal fluid (CSF) findings were retrospectively analyzed from September, 1993 to August, 2006 at Ewha Womans University Mokdong Hospital. Results : Eighty-one cases of bacterial meningitis were observed. Overall the most common organism was Streptococcus agalactiae (group B streptococcus, GBS) (30 cases, 37.0%) followed by Haemophilus influenzae (22 cases, 27.2%), Streptococcus pneumoniae (12 cases, 14.8%), Escherichia coli (3 cases, 3.7%), Neisseria meningitidis (1 case, 1.2%) and others (13 cases, 16.0%). In neonates and young infants under 2 months, the most common organism was GBS. In children between 3 months, and 5 years, the most common organism was H. influenzae. S. pneumoniae was the most common organism in children over 5 years of age. Thirty-one patients (38.3%) had complications. Of all ages, the mortality rate of bacterial meningitis markedly decreased compared with the previously reported rate. Conclusion : In neonates, GBS meningitis was most common. The frequency of H. influenzae meningitis decreased after the introduction of H. influenzae type b vaccination. A strategy for the prevention of GBS meningitis in neonates should be established. The influence of the pneumococcal conjugate vaccine on S. pneumoniae meningitis should be studied.

• Biotechnology and Bioprocess Engineering:BBE
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• v.11 no.3
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• pp.215-222
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• 2006

The Lowry method was used in this study to measure protein in Haemophilus influenzae type b (Hib) conjugate vaccines (polyribosylibitol phosphate-tetanus toxoid; PRP-TT) using deoxycholic acid (DOC) to induce protein precipitation. Trichloroacetic acid (TCA) did not induce precipitation adequately from the Hib conjugate bulk and the freeze-dried Hib conjugate product. Its yield was approximately 50%. The matrix structure of Hib conjugate inhibits precipitation by TCA. Although the Lowry method can be carried out without precipitation in Hib conjugate bulk when no residual impurities (adipic acid dihydrazide [ADH], 1-ethyl-3-(3-dimethylamino-propyl) carbodiimide-HCI [EDAC], phenol and cyanogens bromide [CNBr], etc.) are present, it cannot be used for Hib conjugate products that contain sucrose 8.5%, because 8.5% concentration of sucrose enhanced the protein concentration. DOC- and HCl-induced precipitation is an alternative method for evaluating the protein content of the Hib conjugate bulk and the Hib conjugate product. The precipitation was optimal with a final concentrate of 0.1% for DOC at $4^{\circ}C$ and pH 2. This Lowry method, using DOC/HCI precipitation to induce protein precipitation, was confirmed a consistent, reproducible, and valid test for proteins in Hib conjugate bulk and its freeze-dried product.

• Pediatric Infection and Vaccine
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• v.6 no.1
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• pp.53-68
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• 1999

• Pediatric Infection and Vaccine
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• v.10 no.2
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• pp.208-214
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• 2003

Purpose : We performed a retrospective study on bacterial meningitis in children, pertaining to the causative organisms, bacterial resistance to antibiotics and the effect of recent introduction of Haemophilus influenzae type b(Hib) vaccine. Methods : We analysed the forty-three cases of bacterial meningitis which had been treated at the Inha University Hospital from June 1996 to June 2003. Results : Nineteen cases(44.2%) of them were infants younger than 2 months of age, and 29 cases(67.5%) younger than 1 year of age. The common causative organisms under 2 months of age were group B streptococcus(GBS)(47.4%), E. coli(21.1%), and Klebsiella pneumoniae(21.1%). In the age group beyond 2 months of age, S. pneumoniae were seen in 50 %, H. influenzae in 16.7% and N. meningitidis in 16.7%. All of the five cases of Hib meningitis had not been vaccinated for Hib. There has been no Hib meningitis cases since 2001. Overall fatality rate was 4.5%, and complication occurred in 39%. Complications was significantly less frequent in patients resumed to be treated within 48 hours after onset compared to after 48 hours after onset. Penicillin resistance of S. pneumoniae and GBS isolated among bacterial meningitis cases was high. Conclusion : Timing of treatment after the onset of the disease appeared the most important factor for prognosis of bacterial meningitis. The cases of H. influenzae meningitis have decreased probably due to Hib vaccination.

• Bulletin of the Korean Chemical Society
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• v.32 no.8
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• pp.2549-2552
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• 2011

In this study, the kinetic properties of wild-type and C117D mutant H. influenzae MurA (Hi MurA), which catalyzes the first reaction in the biosynthetic pathway of the cell wall, were characterized. Purified recombinant Hi MurA was active at pH values ranging from pH 5.5 to pH 10, and its $K_m$ (UNAG), $K_m$ (PEP), and $k_{cat}$ values were measured to be 31 ${\mu}M$, 24 ${\mu}M$, and 210 $min^{-1}$, respectively. Hi MurA activity was effectively inhibited by fosfomycin with an $IC_{50}$ value of 60 ${\mu}M$. Hi MurA contains a cysteine residue (C117) at the loop region near the PEP binding, whereas MurA from fosfomycin resistant Mycobaterium tuberculosis or Chlamydia trachomatis contain an aspartate residue instead of the cysteine at the corresponding site. Aspartate substitution of Cys117 in Hi MurA shifted its optimum pH from 7.8 to 6.0. In addition, the $K_m$ values for UNAG and PEP were increased to 160 ${\mu}M$ and 150 ${\mu}M$, respectively, and the $k_{cat}$ value was significantly reduced to 41 $min^{-1}$. Furthermore, the C117D mutant form of Hi MurA was not inhibited by 1 mM fosfomycin. These results indicate that the Cys117 of Hi MurA is the binding site of fosfomycin and plays an important role in the fast turnover of the catalytic reaction.

• Pediatric Infection and Vaccine
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• v.7 no.2
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• pp.233-239
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• 2000

Purpose : A great deal of youngsters suffer from otitis media, for which antimicrobials are frequently prescribed. Increased antimicrobial resistance forces physicians to judiciously use antimicrobial agents in treating patients with acute otitis media. There have however been few references with regard to otitis media in Korean children, and authors proceeded investigation to look for the causative agents of otitis media in Korean children and their antimicrobial susceptibility. Methods : The study included 65 patients younger than 15 years old who had been cared at the department of pediatrics and otolaryngology in Hanyang University Hospital from July 1994 to June 1999, and diagnosed of otitis media with otorrhea which contained microorganisms isolated in otorrhea culture. The medical records were reviewed for demographic data, isolated organisms and their antimicrobial susceptibility. Results : Among 65 patients, 37(57%) were boys and 28(43%) girls. Distribution of the patients was reciprocal to the age of the patients; 27 patients(41.5%) were younger than 1 year old, 24(36.9%) were 1 to 3 years old with the average of 2.9 years of age. Staphylococus aureus was isolated in 32 patients(49.2%), Streptococcus pneumoniae in 19 patients(29.2%) Haemophilus influenzae in 9 patients(13.8%), Streptococcus oralis in 3 patients(4.6%), Moraxella catarrhalis in 1 patient(1.5%). The isolated microorganisms were not different whether patients had cleft lip/palate or not. The antibiotic resistance rates of S. aureus were ${\geq}90%$ to erythromycin, imipenem, cephalothin, and clindamycin, 86.2% to oxacillin, 25% to chloramphenicol, 12.5% to trimethoprim/sulfamethoxazole(TMP/SMX), and 0% to vancomycin and teicoplanin. The antibiotic resistance rates of S. pneumoniae were 71.4% to penicillin and greater than 60% to erythromycin, tetracycline, TMP/SMX, 7.1% to chloramphenicol, and 0% to vancomycin and teicoplanin. The antibiotic resistance rates of H. influenzae were 55% to ampicillin and TMP/SMX, and 0% to chloramphenicol, ceftriaxone, aztreonam, imipenem and ciprofloxacin. Conclusion : With otorrhea culture, the causative organisms of otitis media appear to be S. aureus, S. pneumoniae and H. influenzae. The high antibiotic resistance rates of the isolated organisms should affect the choice of antibiotics in treating patients with otitis media. Prospective investigations utilizing tympanocentesis in microbiologic studies are needed.

• BMB Reports
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• v.33 no.6
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• pp.514-518
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• 2000

Two open reading frames of Haemophilus influenzae, HI0572 and HI0751, showing homology to a yeast thioredoxin peroxidase II (TPx II) and an E. coli thiol peroxidase $P_{20}$, respectively, were cloned and expressed in E. coli, and then the proteins were subsequently purified and characterized. HI0751 protein showed the thioredoxin (Trx)-dependent peroxidase activity, whereas HI0572 protein showed glutathione-dependent peroxidase. The HI0572 is the first peroxiredoxin with glutathione peroxidase activity rather than thioredoxin peroxidase. Purified HI0572 and HI0751 proteins protected specifically the inactivation of glutamine synthetase by metal catalyzed oxidation (MCO) systems composed of $Fe^{3+}$, $O_2$ and mercaptans such as dithiothreitol, ${\beta}-mercaptoethanol$ and glutathione (GSH). Unlike the HI0751 protein, the HI0572 protein was more effective in protecting glutamine synthetase from inactivation by the $GSH/Fe^{3+}/O_2$ system. It seems that these unique properties of the HI0572 protein are due to the structure containing a glutaredoxin domain at it's C-terminal in addition to a peroxiredoxin domain.