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1.
Article in English | WPRIM | ID: wpr-116877

ABSTRACT

PURPOSE: Escherichia coli sequence type (ST) 131, a multidrug-resistant clone causing extraintestinal infections, has rapidly become prevalent worldwide. However, the epidemiological and clinical features of pediatric infections are poorly understood. We aimed to explore the characteristics of ST131 Escherichia coli isolated from Korean children with urinary tract infections. METHODS: We examined 114 uropathogenic E. coli (UPEC) isolates from children hospitalized at Chung-Ang University Hospital between 2011 and 2014. Bacterial strains were classified into STs by partial sequencing of seven housekeeping genes (adk, fumC, gyrB, icd, mdh, purA, and recA). Clinical characteristics and antimicrobial susceptibility were compared between ST131 and non-ST131 UPEC isolates. RESULTS: Sixteen UPEC isolates (14.0%) were extended-spectrum β-lactamase (ESBL)-producers; 50.0% of ESBL-producers were ST131 isolates. Of all the isolates tested, 13.2% (15 of 114) were classified as ST131. There were no statistically significant associations between ST131 and age, sex, or clinical characteristics, including fever, white blood cell counts in urine and serum, C-reactive protein, radiologic abnormalities, and clinical outcome. However, ST131 isolates showed significantly lower rates of susceptibility to cefazolin (26.7%), cefotaxime (40.0%), cefepime (40.0%), and ciprofloxacin (53.3%) than non-ST131 isolates (65.7%, 91.9%, 92.9%, and 87.9%, respectively; P<0.001 for all). ESBL was more frequently produced in ST131 (53.3%) than in non-ST131 (8.1%) isolates (P<0.01). CONCLUSION: ST131 E. coli isolates were prevalent uropathogens in children at a single medical center in Korea between 2011 and 2014. Although ST131 isolates showed higher rates of antimicrobial resistance, clinical presentation and outcomes of patients were similar to those of patients infected with non-ST131 isolates.


Subject(s)
C-Reactive Protein , Cefazolin , Cefotaxime , Child , Ciprofloxacin , Clone Cells , Escherichia coli , Fever , Genes, Essential , Humans , Korea , Leukocyte Count , Multilocus Sequence Typing , Urinary Tract Infections , Urinary Tract , Uropathogenic Escherichia coli
2.
Article in English | WPRIM | ID: wpr-228230

ABSTRACT

The rotavirus nonstructural glycoprotein, NSP4, has been identified as the first viral enterotoxin capable of inducing diarrhea. To investigate the biological function of NSP4 in the inflammatory process, a cDNA from human rotavirus (Wa strain) RNA segment 10 was amplified by RT-PCR, cloned into TA vector, and subsequently subcloned into pET23b expression plasmid. The expression of NSP4 protein was determined by SDS-PAGE and Western blotting, then, the protein was purified by affinity chromatography on Ni-NTA-agarose column. The inflammatory effects of NSP4, namely, production of nitric oxide (NO), pro-inflammatory cytokines (IL-1β, IL-6, IL-10, and TNF-α), and prostaglandin E2 (PGE₂), was evaluated using NSP4-stimulated RAW 264.7 murine macrophages and compared with those observed after stimulation with lipopolysaccharide (LPS). The levels of IL-1β, IL-6, and TNF-α were significantly increased, and those of NO and PGE₂ also increased in NSP4-stimulated RAW 264.7 cells. These findings indicate that NSP4 plays an important role in the inflammatory response observed during rotavirus infection.


Subject(s)
Blotting, Western , Chromatography, Affinity , Clone Cells , Cytokines , Diarrhea , Dinoprostone , DNA, Complementary , Electrophoresis, Polyacrylamide Gel , Enterotoxins , Glycoproteins , Humans , Inflammation , Interleukin-10 , Interleukin-6 , Macrophages , Nitric Oxide , Plasmids , RNA , Rotavirus Infections , Rotavirus
3.
Article in English | WPRIM | ID: wpr-727830

ABSTRACT

The anti-inflammatory, antioxidant, and antimicrobial properties of artemisinin derived from water, methanol, ethanol, or acetone extracts of Artemisia annua L. were evaluated. All 4 artemisinin-containing extracts had anti-inflammatory effects. Of these, the acetone extract had the greatest inhibitory effect on lipopolysaccharide-induced nitric oxide (NO), prostaglandin E2 (PGE2), and proinflammatory cytokine (IL-1beta , IL-6, and IL-10) production. Antioxidant activity evaluations revealed that the ethanol extract had the highest free radical scavenging activity, (91.0+/-3.2%), similar to alpha-tocopherol (99.9%). The extracts had antimicrobial activity against the periodontopathic microorganisms Aggregatibacter actinomycetemcomitans, Fusobacterium nucleatum subsp. animalis, Fusobacterium nucleatum subsp. polymorphum, and Prevotella intermedia. This study shows that Artemisia annua L. extracts contain anti-inflammatory, antioxidant, and antimicrobial substances and should be considered for use in pharmaceutical products for the treatment of dental diseases.


Subject(s)
Acetone , Aggregatibacter actinomycetemcomitans , alpha-Tocopherol , Artemisia annua , Dinoprostone , Ethanol , Fusobacterium nucleatum , Interleukin-6 , Methanol , Nitric Oxide , Pharmaceutical Preparations , Prevotella intermedia , Stomatognathic Diseases , Water
4.
Article in English | WPRIM | ID: wpr-11595

ABSTRACT

PURPOSE: We investigated the molecular types of uropathogenic Escherichia coli (UPEC) by using conventional phylogrouping, multilocus sequence typing (MLST), and fimH genotyping. METHODS: Samples of patients younger than 18 years of age were collected from the Chung-Ang University Hospital over 2 years. Conventional phylogenetic grouping for UPEC strains was performed by polymerase chain reaction (PCR). Bacterial strain sequence types (STs) were classified on the basis of the results of partial sequencing of seven housekeeping genes. In addition, we analyzed nucleotide variations in a 424-base pair fragment of fimH, a major virulence factor in UPEC. RESULTS: Sixty-four UPEC isolates were analyzed in this study. Phylogenetic grouping revealed that group B2 was the most common type (n=54, 84%). We identified 16 distinctive STs using MLST. The most common STs were ST95 (35.9%), ST73 (15.6%), ST131 (12.5%), ST69 (7.8%), and ST14 (6.3%). Fourteen fimH allele types were identified, of which 11 had been previously reported, and the remaining three were identified in this study. f1 (n=28, 45.2%) was found to be the most common allele type, followed by f6 and f9 (n=7, 11.3% each). Comparative analysis of the results from the three different molecular typing techniques revealed that both MLST and fimH typing generated more discriminatory UPEC types than did PCR-based phylogrouping. CONCLUSION: We characterized UPEC molecular types isolated from Korean children by MLST and fimH genotyping. fimH genotyping might serve as a useful molecular test for large epidemiologic studies of UPEC isolates.


Subject(s)
Alleles , Child , Epidemiologic Studies , Genes, Essential , Humans , Molecular Typing , Multilocus Sequence Typing , Phylogeny , Polymerase Chain Reaction , Urinary Tract Infections , Uropathogenic Escherichia coli , Virulence
5.
Article in English | WPRIM | ID: wpr-728530

ABSTRACT

Recently, Cynanchi wilfordii Radix has gained wide use in Asian countries as a functional food effective for relieving fatigue, osteoporosis, and constipation, particularly in menopausal disorders. However, its anti-inflammatory and anti-microbial activities have not been explored in detail to date. The anti-inflammatory, antioxidant, and anti-bacterial properties of the Cynanchi wilfordii Radix extracts obtained with water, methanol, ethanol, and acetone were compared. All 4 polyphenol-containing extracts exhibited anti-inflammatory and antioxidant effects. The ethanol extract was found to elicit the most potent reduction of nitric oxide (NO), prostaglandin E2 (PGE2), and cytokine (IL-1beta, IL-6, IL-10, and TNF-alpha) levels, as well as inhibit the expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) in a concentration-dependent manner. The evaluation of antioxidant activity also revealed the ethanol extract to have the highest free radical scavenging activity, measured as 85.3+/-0.4%, which is equivalent to 99.9% of the activity of alpha -tocopherol. In the assessment of anti-bacterial activity, only ethanol extract was found to inhibit the growth of the Bacillus species Bacillus cereus and Bacillus anthracis. These results show that polyphenols of Cynanchi wilfordii Radix have anti-inflammatory, antioxidant, and anti-bacterial properties that can be exploited and further improved for use as a supplementary functional food, in cosmetics, and for pharmaceutical purposes.


Subject(s)
Acetone , Antioxidants , Asian Continental Ancestry Group , Bacillus , Bacillus anthracis , Bacillus cereus , Constipation , Cyclooxygenase 2 , Dinoprostone , Ethanol , Fatigue , Functional Food , Humans , Interleukin-10 , Interleukin-6 , Methanol , Nitric Oxide , Nitric Oxide Synthase Type II , Osteoporosis , Polyphenols , Water
6.
Article in English | WPRIM | ID: wpr-218818

ABSTRACT

The MinION(TM) is a miniature nanopore-based analysis device in which the characteristics of an analyte, as it passes through the nanopore, cause changes in the flow of ions through the pore, which are measured, as current flow, by a low noise amplifier and analogue-to-digital converter. Potentially any molecular analyte capable of passing through the nanopore may modify the flow of ions and generate a signal which might be diagnostic. In practice the current device is focussed on DNA sequencing, directly sequencing RNA is a likely development. With the MinION Access Program making the MinION(TM) widely available a flood of applications exploiting its real time, long read capabilities have been published. We review the background to the technology and compare it to current next generation sequencing.


Subject(s)
Ions , Nanopores , Noise , RNA , Sequence Analysis, DNA
7.
Article in English | WPRIM | ID: wpr-727598

ABSTRACT

Shepherd's purse, Capsella bursa-pastoris (L.) Medik., has been considered a health food for centuries in Asia and is known to contain the isothiocyanate compound sulforaphane. In this study, we evaluated the anti-inflammatory and antibacterial properties of a sulforaphane-containing solution (SCS) isolated from shepherd's purse. SCS had significant anti-inflammatory activity indicated by the decreased levels of nitric oxide (NO), cytokines (interleukin 1beta [IL-1beta], IL-6, and IL-10), and prostaglandin E2 (PGE2) in lipopolysaccharide-stimulated RAW 264.7 murine macrophages. In addition, SCS decreased the inducible NO synthase (iNOS) and cyclooxygenase 2 (COX-2) levels, which confirmed the anti-inflammatory activity of SCS. Further, SCS inhibited vancomycin-resistant enterococci (VRE) and Bacillus anthracis. The minimal inhibitory concentration was 250 microg/ml for VRE and 1,000 microg/ml for B. anthracis. Taken together, these data indicate that SCS has potential anti-inflammatory and anti-superbacterial properties, and thus it can be used as a functional food or pharmaceutical.


Subject(s)
Asia , Bacillus anthracis , Capsella , Cyclooxygenase 2 , Cytokines , Dinoprostone , Functional Food , Food, Organic , Interleukin-6 , Macrophages , Nitric Oxide , Nitric Oxide Synthase
8.
Article in English | WPRIM | ID: wpr-70870

ABSTRACT

Hepatitis A virus (HAV) positive stool samples were collected from acute hepatitis A patients during the two study periods of 2002 and 2011 in Seoul, South Korea, and their genetic variability was determined. From a total of 79 specimens, the nucleotide sequences of the VP1 and 2A junction were successfully amplified in 27 (34.2%) samples and subjected to sequence analysis. Genetically, there was a dramatic change in HAV subgenotypes from IA to IIIA during the past ten years. Sequence analysis identified that most strains belonged to genotype I, which is the main genotype globally. The subgenotype IA (93.3%, n=13/14) was the major subgenotype in 2002, whereas the subgenotype IIIA (69.2%, n=9/13) was predominant in 2011. Interestingly, a IIIA strain was identified from a patient who had a history of travel to India in 2002. The finding presented provides new insight into the genetic shift of circulating HAVs in South Korea.


Subject(s)
Base Sequence , Genetic Variation , Genotype , Hepatitis A , Hepatitis A virus , Humans , India , Korea , Seoul , Sequence Analysis
9.
Article in English | WPRIM | ID: wpr-188345

ABSTRACT

To observe how anti-group A rotavirus antibody seropositivity rates and levels have changed in the western region of Gyeongnam Province, 2,030 serum samples collected at four collection periods (1989-1990, 1994-1995, 1999-2000, and 2004-2005) were tested by Enzyme-Linked Immunosorbent Assay for IgG, and IgA antibodies reacting to recombinant VP6 protein. The seroprevalences exhibit no regular patterns over a 16-yr period. For all four collection periods, the anti-rVP6 IgG levels rose steadily during the first 5 months of life, after which they remained high. However, the 2-9 yr and 10-39 yr groups had significantly higher IgG levels in 1999-2000 and 2004-2005, respectively, than in the other collection periods. The 1-5 mo, 40- > or = 60 yr, and 4-29 yr groups had significantly higher IgA levels in 1989-1990, 1999-2000, and 2004-2005, respectively. The 4 yr (25.0%), 5-9 yr (18.8%), 10-14 yr (41.1%), 20-29 yr (35.0%), and 30-39 yr (20.0%) groups in 2004-2005 had significant higher IgA seropositivity rate compared to the other three collection periods. These observations suggest that in the western region of Gyeongnam Province since the late 1990s, rotavirus reinfection has occurred more frequently than previously, with all ages being at risk.


Subject(s)
Adult , Aged , Antibodies, Viral/blood , Antigens, Viral/genetics , Capsid Proteins/genetics , Child , Child, Preschool , Enzyme-Linked Immunosorbent Assay , Female , Humans , Immunoglobulin A/blood , Immunoglobulin G/blood , Infant , Infant, Newborn , Male , Middle Aged , Recombinant Proteins/biosynthesis , Republic of Korea/epidemiology , Rotavirus/isolation & purification , Rotavirus Infections/epidemiology , Seroepidemiologic Studies , Time Factors , Young Adult
10.
Article in English | WPRIM | ID: wpr-727486

ABSTRACT

The fruit of the black raspberry (Rubus coreanus Miquel) has been employed in traditional medicine, and recent studies have demonstrated its measureable biological activities. However, the root of the black raspberry has not been studied. Therefore, in this study, we evaluated the anti-inflammatory and antibacterial properties of the root and unripe fruit polyphenols of the black raspberry. Both polyphenols proved to have anti-inflammatory activity as evidenced by the decreased nitric oxide (NO), cytokines (IL-1beta , IL-6, and IL-10) and prostaglandin E2 (PGE2) levels in lipopolysaccharide (LPS)-stimulated RAW 264.7 murine macrophages. However, root polyphenols showed stronger anti-inflammatory activity than fruit polyphenols. LPS-induced mRNA and protein expressions of inducible NO synthase (iNOS) and cyclooxygenase (COX)-2 levels were also decreased, confirming the anti-inflammatory activity. Root polyphenols showed lethal activity against methicillin-resistant Staphylococcus aureus (MRSA), carbapenem-resistant Acinetobacter baumannii (CRAB), and Bacillus anthracis. In contrast, the black raspberry fruit did not demonstrate these properties. These data provide the first demonstration that black raspberry root has potential anti-inflammatory and anti-superbacterial properties that can be exploited as alternatives for use in the food and cosmetic industries and/or as pharmaceuticals.


Subject(s)
Acinetobacter baumannii , Bacillus anthracis , Cosmetics , Cytokines , Dinoprostone , Fruit , Interleukin-6 , Macrophages , Medicine, Traditional , Methicillin-Resistant Staphylococcus aureus , Nitric Oxide , Nitric Oxide Synthase , Polyphenols , Prostaglandin-Endoperoxide Synthases , RNA, Messenger
11.
Article in English | WPRIM | ID: wpr-30994

ABSTRACT

The epidemiology of human group A rotavirus was analyzed by examining genotypic data acquired from 1989 to 2009 in South Korea. This information was derived from all the available published articles on rotavirus studies in South Korea, retrieved from both the PubMed and KoreaMed databases. Four common G types (G1, G2, G3, and G4) and three common P types (P[8], P[4], and P[6]) accounted for approximately 93% and 99% of the rotavirus reports, respectively. The G9 type was frequently detected after 2000, and because of this prevalence, it is considered to be the fifth most important G type rotavirus after the G1.G4 genotypes. Less common G types of the virus such as G12, G11, and G10 were detected in some geographic settings, and it is important to consider the context of these subtypes and their epidemiological significance. The P[9] virus genotype was observed in the study and has been discussed in many other studies; however, the P[3], P[10] and P[25] genotypes were rarely detected in the epidemiological research. In general, the distributions of the G and P genotypes showed temporal and geographical fluctuations, and a nationwide rotavirus vaccine program that targeted these genotypes demonstrated effectiveness in protecting against the circulating rotavirus strains. However, further analysis is needed to determine the true long-term effectiveness of these vaccines; the analysis should also consider the unexpected effects of vaccinations, such as vaccine-induced diseases, herd immunity, and changes in host susceptibilities.


Subject(s)
Epidemiology , Genotype , Humans , Immunity, Herd , Prevalence , Republic of Korea , Rotavirus , Vaccination , Vaccines , Viruses
12.
Article in English | WPRIM | ID: wpr-97575

ABSTRACT

Bronchial asthma can be triggered by microbial agents in the oropharynx. This study was designed to identify the differences in microbiota of oropharynx of bronchial asthmatic patients in contrast to normal controls. In order to resolve the qualitative and quantitative diversity of the 16S rRNA gene present in the oropharynx microbiota of 4 patients and 4 controls, we compared microbial communities using Sanger sequencing and 376 sequences of 16S rRNA gene were analyzed. Of the total microbial diversity detected in the oropharynx in asthmatic patients 45.6% comprised members of the Firmicutes. In contrast, Proteobacteria (44.0%) dominated the oropharyngeal microbiota in the normal control group. Members of the Bacteroidetes, Fusobacteria, Actinobacteria, TM7, Cyanobacteria and unclassified bacteria were present in both groups. In conclusion, the difference in the microbiota of the oropharynx between patients and normal individuals could trigger symptomatic attacks in bronchial asthma.


Subject(s)
Actinobacteria , Asthma , Bacteria , Bacteroidetes , Cyanobacteria , Fusobacteria , Genes, rRNA , Humans , Metagenome , Microbiota , Oropharynx , Proteobacteria
13.
Article in English | WPRIM | ID: wpr-97573

ABSTRACT

Although viruses are the most common cause of acute gastroenteritis (AGE) in humans, details about the causative viruses in AGE are largely unknown because many causative viruses are unable to be cultured by current culture techniques. In our study, fecal samples from 10 children under five years of age with unexplained AGE and 10 healthy children were investigated for RNA viruses using random priming (RP)-mediated sequence-independent single primer amplification (SISPA). The causative viruses in cases of cryptogenic diarrhea were then assessed for their potential diagnostic value. Of the 1,129 viral clones identified, rotavirus was most commonly associated with AGE (125 sequences, 22.4%). In contrast, bacteriophage was most common (43 sequences, 13.6%) in healthy children. The remaining 515 viral clones were unidentifiable. These findings suggest that investigation of cases or outbreaks of unexplained diarrhea using a metaviromic strategy is a new avenue for diagnosis.


Subject(s)
Bacteriophages , Child , Clone Cells , Culture Techniques , Diagnosis , Diarrhea , Disease Outbreaks , Feces , Gastroenteritis , Humans , Parechovirus , RNA Viruses , Rotavirus
14.
Article in English | WPRIM | ID: wpr-200678

ABSTRACT

Uncultured microorganisms comprise the majority of the planet's biological diversity. In many environments, as many as 99% of the microorganisms cannot be cultured by standard techniques, and the uncultured fraction includes diverse organisms that are only distantly related to the cultured ones. Therefore, culture-independent methods are essential to understand the genetic diversity, population structure, and ecological roles of the majority of microorganisms. Recently, new techniques for studying microbial communities, collectively called metagenomics, have been developed to overcome the limitations of culturing. This review assesses the potential of metagenomic techniques to analyze the relative abundance of microbial species under varying human environmental conditions and to discover infectious causes of unexplained human diseases.


Subject(s)
Biodiversity , Communicable Diseases , Genetic Variation , Humans , Metagenome , Metagenomics
15.
Article in English | WPRIM | ID: wpr-165303

ABSTRACT

The effect of DMSO and sodium butyrate on the production of recombinant hepatitis A virus (HAV) capsid protein VP1 was evaluated and optimized in the culture of stably transfected Drosophila melanogaster S2 cells using culture plates and spinner flasks. The effect of DMSO and sodium butyrate was also evaluated to improve the recombinant VP1 production in stably transfected Drosophila S2 cells. A production level of 0.88 mg of recombinant VP1/liter was obtained in the culture-plate culture of stably transfected S2 cells at 6 days after induction with 0.5 mM CuSO4. The supplements of 2% DMSO and 10 mM sodium butyrate at 4 days post-inoculation increased recombinant VP1 accumulation by 141 and 104%, respectively, resulting in 2.17 and 1.7 mg/liter of recombinant VP1 production. In spinner flasks, recombinant VP1 production reached maximum level at 9 days after induction with 0.5 mM CuSO4, with approximately 4.96 mg/liter of recombinant VP1 production level. When 2% DMSO or 10 mM sodium butyrate was added at 5 days post-inoculation, the recombinant VP1 production was increased to 8.35 and 5.85 mg/liter, respectively. However, the synergistic effects of DMSO and sodium butyrate were not observed. These results indicate that DMSO and/or sodium butyrate can be successfully used to improve the recombinant HAV VP1 production in culture plates and spinner flasks.


Subject(s)
Butyrates , Capsid Proteins , Dimethyl Sulfoxide , Drosophila , Drosophila melanogaster , Efficiency , Hepatitis , Hepatitis A , Hepatitis A virus , Sodium
16.
Article in Korean | WPRIM | ID: wpr-130990

ABSTRACT

PURPOSE: The serologic diagnosis of rotaviral infections is not commonly used in clinical practice, but is used in seroepidemiologic studies. In this study, the usefulness of Escherichia coli-expressed recombinant VP6 proteins of group A rotavirus in the serodiagnosis of rotavirus infections by ELISA was evaluated. METHODS: The recombinant VP6 proteins of group A rotavirus expressed in E. coli Rosetta II strain were purified and identified. One hundred sera from 22 children (4 healthy neonates, 13 healthy children, and 5 immunocompromised children) who had serial sera samples prior to and after rotavirus infections were provided by the Gyeongsang National University Hospital, a member of the National Biobank of Korea. IgG, IgA, and IgM antibodies against rVP6 were analyzed by ELISA in all of the patients and Western blot analysis in 4 neonates. RESULTS: ELISA tests using rVP6 proteins of group A rotavirus as antigen revealed that IgG, IgA, and IgM antibodies increased after rotaviral infections in most neonates and healthy children. IgG antibodies also increased after rotaviral infections in most immunocompromised children without an adequate increase in IgM or IgA antibodies. Western blot analysis in four neonates revealed very early IgM antibody responses, even in the sera with low optical densities in ELISA tests. CONCLUSION: Our study showed that ELISA using rVP6 as an antigen is a valid diagnostic tool for seroepidemiologic studies of rotavirus infections and Western blot analysis is a sensitive test in detecting IgG, IgA, and and IgM antibodies in patients with rotavirus infections.


Subject(s)
Antibodies , Antibody Formation , Blotting, Western , Child , Enzyme-Linked Immunosorbent Assay , Escherichia , Humans , Immunoglobulin A , Immunoglobulin G , Immunoglobulin M , Infant, Newborn , Korea , Proteins , Rotavirus , Rotavirus Infections , Seroepidemiologic Studies , Serologic Tests , Sprains and Strains
17.
Article in Korean | WPRIM | ID: wpr-130987

ABSTRACT

PURPOSE: The serologic diagnosis of rotaviral infections is not commonly used in clinical practice, but is used in seroepidemiologic studies. In this study, the usefulness of Escherichia coli-expressed recombinant VP6 proteins of group A rotavirus in the serodiagnosis of rotavirus infections by ELISA was evaluated. METHODS: The recombinant VP6 proteins of group A rotavirus expressed in E. coli Rosetta II strain were purified and identified. One hundred sera from 22 children (4 healthy neonates, 13 healthy children, and 5 immunocompromised children) who had serial sera samples prior to and after rotavirus infections were provided by the Gyeongsang National University Hospital, a member of the National Biobank of Korea. IgG, IgA, and IgM antibodies against rVP6 were analyzed by ELISA in all of the patients and Western blot analysis in 4 neonates. RESULTS: ELISA tests using rVP6 proteins of group A rotavirus as antigen revealed that IgG, IgA, and IgM antibodies increased after rotaviral infections in most neonates and healthy children. IgG antibodies also increased after rotaviral infections in most immunocompromised children without an adequate increase in IgM or IgA antibodies. Western blot analysis in four neonates revealed very early IgM antibody responses, even in the sera with low optical densities in ELISA tests. CONCLUSION: Our study showed that ELISA using rVP6 as an antigen is a valid diagnostic tool for seroepidemiologic studies of rotavirus infections and Western blot analysis is a sensitive test in detecting IgG, IgA, and and IgM antibodies in patients with rotavirus infections.


Subject(s)
Antibodies , Antibody Formation , Blotting, Western , Child , Enzyme-Linked Immunosorbent Assay , Escherichia , Humans , Immunoglobulin A , Immunoglobulin G , Immunoglobulin M , Infant, Newborn , Korea , Proteins , Rotavirus , Rotavirus Infections , Seroepidemiologic Studies , Serologic Tests , Sprains and Strains
18.
Article in English | WPRIM | ID: wpr-728746

ABSTRACT

Functional defects in mitochondria are involved in the induction of cell death in cancer cells. We assessed the toxic effect of camptothecin against the human cervical and uterine tumor cell line SiHa with respect to the mitochondria-mediated cell death process, and examined the combined effect of camptothecin and anticancer drugs. Camptothecin caused apoptosis in SiHa cells by inducing mitochondrial membrane permeability changes that lead to the loss of mitochondrial membrane potential, decreased Bcl-2 levels, cytochrome c release, caspase-3 activation, formation of reactive oxygen species and depletion of GSH. Combination of camptothecin with other anticancer drugs (carboplatin, paclitaxel, doxorubicin and mitomycin c) or signaling inhibitors (farnesyltransferase inhibitor and ERK inhibitor) did not enhance the camptothecin-induced cell death and caspase-3 activation. These results suggest that camptothecin may cause cell death in SiHa cells by inducing changes in mitochondrial membrane permeability, which leads to cytochrome c release and activation of caspase-3. This effect is also associated with increased formation of reactive oxygen species and depletion of GSH. Combination with other anticancer drugs (or signaling inhibitors) does not appear to increase the anti-tumor effect of camptothecin against SiHa cells, but rather may reduce it. Combination of camptothecin with other anticancer drugs does not seem to provide a benefit in the treatment of cervical and uterine cancer compared with camptothecin monotherapy.


Subject(s)
Apoptosis , Camptothecin , Carcinoma, Squamous Cell , Caspase 3 , Cell Death , Cell Line , Cell Line, Tumor , Cytochromes c , Doxorubicin , Humans , Membrane Potential, Mitochondrial , Mitochondria , Mitochondrial Membranes , Mitomycin , Paclitaxel , Permeability , Reactive Oxygen Species , Uterine Neoplasms
19.
Korean Journal of Medicine ; : 189-197, 2006.
Article in Korean | WPRIM | ID: wpr-91909

ABSTRACT

BACKGROUND: The aims of this study were to compare the suitability of repetitive-PCR genomic fingerprinting procedures to investigate genetic relatedness of the genus Vibrio and its applicability for the molecular identification of Vibrio vulnificus. METHODS: Forty-eight Vibrio strains were included for this study. REP-, ERIC-, BOX- and SERE-PCR were compared with 13 members of the genus Vibrio. RESULTS: REP-, BOX- and SERE-PCR showed V. vulnificus strains could not be separated well from other Vibrio species. However, approximately 320 bp of highly discriminatory specific fragments was recovered from V. vulnificus strains by ERIC-PCR. CONCLUSIONS: ERIC-PCR could be used as rapid classification and identification methods of V. vulnificus from other members of the genus Vibrio.


Subject(s)
Classification , Dermatoglyphics , Vibrio vulnificus , Vibrio
20.
Article in Korean | WPRIM | ID: wpr-83365

ABSTRACT

PURPOSE: Efficacy of the new rotavirus vaccines (Rotarix(R), RotaTeq(R)) recently developed can be affected by the rotavirus genotypes prevalent in communities. We performed this study to identify the recent distribution of rotavirus genotypes prevalent in Jeju. METHODS: Genotyping of human rotaviruses was performed using 81 samples collected from 154 inpatients and outpatients with rotavirus gastroenteritis at Cheju National University Hospital between July 2005 and June 2006. All six (1, 2, 3, 4, 8, 9) G serotypes were identified by amplification of segments of the gene for VP7 using the reverse transcription-polymerase reaction (RT- PCR). RESULTS: The results of RT-PCR for 81 samples were all positive. G typing of the VP7 protein showed that G1 was the most dominant circulating genotype (65.5%) followed by G2 (14.8%), G3 (13.6%), G8 (1.2%), G9 (1.2%), G4 (0%), and a combination of G1/G3 (3.7%). CONCLUSION: This distribution of rotavirus VP7 genotypes in Jeju is different from that in other domestic areas; the most dominant circulating genotype was G1.


Subject(s)
Gastroenteritis , Genotype , Humans , Infant , Inpatients , Outpatients , Rotavirus Vaccines , Rotavirus
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