1.Purification of recombinant lysostaphin by monoclonal antibody affinity chromatography.
Jinjiang HUANG ; Hongyu WU ; Jien ZHANG ; Qingshan HUANG
Chinese Journal of Biotechnology 2009;25(1):147-151
Lysostaphin, a specific endopeptidase enzyme derived from Staphylococcus aureus, is a bactericidal agent against Staphylococcus and difficult to be drug-resistant. This study established the monoclonal antibody affinity chromatography to obtain lysostaphin of high purity for drug-use standard. The purified Lysostaphin was of > 95% purity and its recovery rate more than 90%. Moreover, the affinity column kept its efficiency of purification invariable after more than 30 times repeat. Also, the dye release assay validated that the purified lysostaphin had significant bactericidal activity. This method was simple and of high efficacy for the lysostaphin purification and showed its potency in commercial production.
Antibodies, Monoclonal
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immunology
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Chromatography, Affinity
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methods
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Lysostaphin
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biosynthesis
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isolation & purification
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Recombinant Proteins
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biosynthesis
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isolation & purification
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Staphylococcus aureus
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enzymology
2.Repeated Serum Alkaline Phosphatase Measurements in the Treatment of Childhood Acute Bone and Joint Infections with High Doses of Antibiotics.
Markus PAAKKONEN ; Markku J T KALLIO ; Pentti E KALLIO ; Heikki PELTOLA
Annals of Laboratory Medicine 2013;33(5):375-378
No abstract available.
Acute Disease
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Adolescent
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Alkaline Phosphatase/*blood
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Anti-Bacterial Agents/*therapeutic use
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Arthritis, Infectious/*drug therapy/*enzymology/microbiology
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*Bacterial Infections/drug therapy/enzymology/microbiology
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Child
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Child, Preschool
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Haemophilus influenzae type b/isolation & purification
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Humans
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Infant
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Osteomyelitis/*drug therapy/*enzymology/microbiology
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Staphylococcus aureus/isolation & purification
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Streptococcus pneumoniae/isolation & purification
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Streptococcus pyogenes/isolation & purification
3.Coagulase-Positive Staphylococcal Necrotizing Fasciitis Subsequent to Shoulder Sprain in a Healthy Woman.
Hyeung June KIM ; Dong Heon KIM ; Duk Hwan KO
Clinics in Orthopedic Surgery 2010;2(4):256-259
Necrotizing fasciitis (NF) is a deep infection of the subcutaneous tissue that progressively destroys fascia and fat; it is associated with systemic toxicity, a fulminant course, and high mortality. NF most frequently develops from trauma that compromises skin integrity, and is more common in patients with predisposing medical conditions such as diabetes mellitus, atherosclerosis, alcoholism, renal disease, liver disease, immunosuppression, malignancy, or corticosteroid use. Most often, NF is caused by polymicrobial pathogens including aerobic and anaerobic bacteria. NF caused by Staphylococcus aureus as a single pathogen, however, is rare. Here we report a case of NF that developed in a healthy woman after an isolated shoulder sprain that occurred without breaking a skin barrier, and was caused by Staphylococcus aureus as a single pathogen.
*Arm
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Coagulase/metabolism
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Fasciitis, Necrotizing/*etiology/microbiology/pathology/surgery
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Female
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Humans
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Middle Aged
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Shoulder Joint/*injuries
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Sprains and Strains/*complications
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Staphylococcal Infections/*etiology/microbiology
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Staphylococcus aureus/enzymology/isolation & purification
4.Evaluation of BD MAX Staph SR Assay for Differentiating Between Staphylococcus aureus and Coagulase-Negative Staphylococci and Determining Methicillin Resistance Directly From Positive Blood Cultures.
Jaewoong LEE ; Yeon Joon PARK ; Dong Jin PARK ; Kang Gyun PARK ; Hae Kyung LEE
Annals of Laboratory Medicine 2017;37(1):39-44
BACKGROUND: We evaluated the performance of the BD MAX StaphSR Assay (SR assay; BD, USA) for direct detection of Staphylococcus aureus and methicillin resistance not only in S. aureus but also in coagulase-negative Staphylococci (CNS) from positive blood cultures. METHODS: From 228 blood culture bottles, 103 S. aureus [45 methicillin-resistant S. aureus (MRSA), 55 methicillin-susceptible S. aureus (MSSA), 3 mixed infections (1 MRSA+Enterococcus faecalis, 1 MSSA+MRCNS, 1 MSSA+MSCNS)], and 125 CNS (102 MRCNS, 23 MSCNS) were identified by Vitek 2. For further analysis, we obtained the cycle threshold (Ct) values from the BD MAX system software to determine an appropriate cutoff value. For discrepancy analysis, conventional mecA/mecC PCR and oxacillin minimum inhibitory concentrations (MICs) were determined. RESULTS: Compared to Vitek 2, the SR assay identified all 103 S. aureus isolates correctly but failed to detect methicillin resistance in three MRSA isolates. All 55 MSSA isolates were correctly identified by the SR assay. In the concordant cases, the highest Ct values for nuc, mecA, and mec right-extremity junction (MREJ) were 25.6, 22, and 22.2, respectively. Therefore, we selected Ct values from 0-27 as a range of positivity, and applying this cutoff, the sensitivity/specificity of the SR assay were 100%/100% for detecting S. aureus, and 97.9%/98.1% and 99.0%/95.8% for detecting methicillin resistance in S. aureus and CNS, respectively. CONCLUSIONS: We propose a Ct cutoff value for nuc/mec assay without considering MREJ because mixed cultures of MSSA and MRCNS were very rare (0.4%) in the positive blood cultures.
Anti-Bacterial Agents/pharmacology
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Bacteremia/diagnosis/microbiology
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Coagulase/metabolism
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Humans
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Methicillin-Resistant Staphylococcus aureus/drug effects/genetics/*isolation & purification
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Microbial Sensitivity Tests
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Oxacillin/pharmacology
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Reagent Kits, Diagnostic
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Staphylococcus/drug effects/enzymology/genetics/isolation & purification
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Staphylococcus aureus/drug effects/genetics/*isolation & purification
5.Active compounds from rhizomes of Musa basjoo.
China Journal of Chinese Materia Medica 2010;35(18):2424-2427
OBJECTIVETo study the active compounds from the rhizomes of Musa basjoo.
METHODAntioxidant and alpha-glucosidase inhibitory activity of different extracts were tested. Using bioassay-guided fractionation, the chemical constituents in EtOAC extracts were isolated by column chromatography and identified by MS and NMR spectroscopy.
RESULTFive compounds were isolated and identified as 2',3, 4'-trihydroxyflavone (1), 3,3'-bis-hydroxyanigorufone (2), irenolone (3), 4-dihydroxy-9-(4'-hydroxyphenyl)-phenalenone (4) and 3,4-dihydroxybenzaldehyde (5). Compound 1(IC50 8.61 mg x L(-1)), 3 (IC50 19.55 mg x L(-1)) and 5 (IC50 1.1 mg x L(-1)) had antioxidant activity. Compound 2 (IC50 24.15 mg x L(-1)) and 4(IC50 2.81 mg x L(-1)) had alpha-glucosidase inhibitory activity. Compound 5 showed MIC of 0.078, 0.313, 0.039 microg/disc against SA, MRSA and ESBLs, respectively.
CONCLUSIONCompound 1-5 were isolated from this plant for the first time. Compound 5 was isolated from the genus Musa for the first time. All compound except 5 were first reported about activity.
Bacterial Proteins ; analysis ; antagonists & inhibitors ; Enzyme Inhibitors ; analysis ; isolation & purification ; pharmacology ; Glycoside Hydrolase Inhibitors ; Musa ; chemistry ; Plant Extracts ; analysis ; isolation & purification ; pharmacology ; Rhizome ; chemistry ; Staphylococcus aureus ; drug effects ; enzymology ; alpha-Glucosidases ; analysis
6.Blood Culture Proven Early Onset Sepsis and Late Onset Sepsis in Very-Low-Birth-Weight Infants in Korea.
Soon Min LEE ; Meayoung CHANG ; Ki Soo KIM
Journal of Korean Medical Science 2015;30(Suppl 1):S67-S74
Neonatal sepsis remains one of the most important causes of death and co-morbidity in very-low-birth-weight (VLBW) infants. The aim of this study was to determine the current incidences of early-onset sepsis (EOS) and late-onset sepsis (LOS), the distribution of pathogens, and the impact of infection on co-morbidities in VLBW infants. We analyzed the data including sepsis episode from 2,386 VLBW infants enrolled in Korean Neonatal Network from January 2013 to June 2014. We defined EOS as a positive blood culture occurring between birth and 7 days of life and LOS after 7 days of life. Sepsis was found in 21.1% of VLBW infants. The risk of sepsis was inversely related to birth weight and gestational age. EOS was found in only 3.6% of VLBW infants, however the mortality rate was as high as 34.1%. EOS was associated with the increased odds for bronchopulmonary dysplasia and intraventricular hemorrhage. The vast majority of EOS was caused by Gram-positive organisms, particularly coagulase-negative staphylococci (30.6%). LOS developed in 19.4% of VLBW infants with a 16.1% mortality rate. Pathogens in LOS were dominated by coagulase-negative staphylococci (38.3%). Twenty-five percent and fifty percent of first LOS episode occurred after 12 days and 20 days from birth, respectively. Younger and smaller VLBW infants showed the earlier occurrence day for the 25% of first LOS episode. This study provides a recent nationwide epidemiology of sepsis in VLBW infants in Korea. Based on this study, successful strategies to reduce infections would improve survival and reduce morbidity.
Coagulase/metabolism
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Databases, Factual
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Gestational Age
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Gram-Negative Bacteria/isolation & purification
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Gram-Positive Bacteria/isolation & purification
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Humans
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Incidence
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Infant, Newborn
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*Infant, Very Low Birth Weight
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Kaplan-Meier Estimate
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Republic of Korea/epidemiology
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Risk Factors
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Sepsis/*epidemiology/microbiology/mortality
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Staphylococcus/enzymology/isolation & purification