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Identification of Proteome Molecules by Proteomics Using Two-Dimensional Gel Electrophoresis and MALDI-TOF MS.

Korean Journal of Infectious Diseases.2001;33(6):470-476.

Genomic technologies have enabled rapid accumulation of information from complex biological systems over the last two decades. The complete DNA sequence is now known for many organism and the informational database obtained from genome sequencing projects has provide the base for the specification of proteome-the protein complement of genome. Genomic functions can be inferred from the analysis of gene structure and gene expression profiles because proteins are the functional molecules of an organism. Integrated technologies including protein separation, identification, characterization and information manage system are essential to analyze the proteins in complex cellular matrix. This study is focusing on the strategies of proteome analysis using sample preparation, 2-dimensional gel electrophoresis, processing of protein spots and identification of proteins, protein-protein interaction and posttranslational modification using MALDI-TOF-MS. 2-Dimensional gel electrophoresis is currently the most powerful protein separation technique and MALDI-TOF MS is powerful identification technique for protein and peptides as a sensitive, rapid, and high resolution analytical method. The developed integrated proteome technologies are very useful to understand the biological phenomena at molecular level by identifying the new molecules and their modifications in various cellular processes, and can be applied for biotechnology including medical science.
Base Sequence ; Biological Phenomena ; Biotechnology ; Complement System Proteins ; Electrophoresis ; Electrophoresis, Gel, Two-Dimensional* ; Genome ; Peptides ; Protein Processing, Post-Translational ; Proteome* ; Proteomics* ; Transcriptome

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Scrub Typhus in a Pregnant Woman: No Evidence of Intrauterine Infection.

Ji Hwan BANG ; Young Ju CHOE ; Yo Han JOH ; Ui Seok KIM ; Jong Wook SHIN ; Hang Rae KIM ; Myoung Don OH ; Ik Sang KIM ; Kang Won CHOE

Korean Journal of Infectious Diseases.2001;33(6):453-455.

Pregnancy with scrub typhus is a rare condition. A 30-year-old woman was infected with scrub typhus at the 35th week of gestation. She was treated successfully with azithromycin, and delivered her baby uneventfully. The baby developed no signs for scrub typhus, and thrived well. IgM antibodies to O. tsutsugamushi were undetectable in the baby's sera, and titers of IgG antibodies did not rise. The polymerase chain reaction of the cord blood for O. tsutsugamushi was also negative. We concluded that transplacental infection did not occur in this pregnant woman.
Adult ; Antibodies ; Azithromycin ; Female ; Fetal Blood ; Humans ; Immunoglobulin G ; Immunoglobulin M ; Polymerase Chain Reaction ; Pregnancy ; Pregnant Women* ; Scrub Typhus*

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Human Herpes Virus-6 Meningoencephalitis Following Allogeneic Hematopoietic Stem Cell Transplantation.

Dong Gun LEE ; Yoo Jin KIM ; Jung Hyun CHOI ; Wan Shik SHIN ; Sang Tae PARK ; Dong Kyun SON ; Dong Wook KIM ; Woo Sung MIN ; Chun Choo KIM

Korean Journal of Infectious Diseases.2001;33(6):448-452.

Two cases of human herpesvirus-6 (HHV-6) encephalitis that occurred after allogeneic hematopoietic stem cell transplant (HSCT) are presented. Both patients reported fever, skin rashes, and neurological symptoms, characterized by disorientation, confusion and drowsy mental status. HHV-6B DNA was detected from the cerebrospinal fluid (CSF) specimens by a multiplex polymerase chain reaction (PCR) assay including EBV, CMV, HHV-6B. After treatment with intravenous gan ciclovir for 2~3 weeks, all clinical manifestations were resolved and the HHV-6B DNA was cleared from the CSF in both patients. HHV-6 should be considered as a possible cause of neurological symptoms after HSCT, and prompt antiviral treatment should be begun.
Bone Marrow Transplantation ; Cerebrospinal Fluid ; DNA ; Encephalitis ; Exanthema ; Fever ; Hematopoietic Stem Cell Transplantation* ; Hematopoietic Stem Cells* ; Herpesvirus 4, Human ; Herpesvirus 6, Human ; Humans* ; Meningoencephalitis* ; Multiplex Polymerase Chain Reaction

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Epidemiologic and Clinical Features of Adult Patients with Measles During 2000 Epidemic.

Joon Young SONG ; Cheong Won PARK ; Jong Seop LEE ; Joong Shik EOM ; Jang Wook SOHN ; Hee Jin CHEONG ; Woo Joo KIM ; Min Ja KIM ; Seung Chull PARK

Korean Journal of Infectious Diseases.2001;33(6):443-447.

BACKGROUND: Measles is an acute highly contagious infection caused by measles virus. Since the introduction of measles vaccines in 1963, measles has been dramatically decreased in Korea. However, measles outbreaks have occurred periodically every 4 to 6 years during the 1990s. During 2000, measles outbreak of large scale occurred throughout the country and the total of 32,088 cases were reported. The majority of affected population was infants and school-aged children, but measles was reported also among adolescent and adult population. There was no report about the epidemiology of adult measles cases during the last decade. In this study, we retrospectively analyzed the epidemiologic and clinical features of adult measles patients in Korea. METHODS: From July of 2000 to January of 2001, 50 adult cases of measles had been admitted to Korea University Hospital. Medical records of these 50 patients, who were clinically diagnosed as measles, were reviewed. Demographic data, clinical features, history of measles vaccination and laboratory findings were evaluated. RESULTS: The median age of patients was 23 years : 28 (56%) were male. Most cases (49/50, 98%) occurred between November of 2000 to January of 2001 and 8 (16%) patients were associated with outbreaks. The occupation of patients was diverse but, students (28%) and office workers (20%) were the most common. 44 (88%) of 50 patients had no underlying diseases. Adult measles patients demonstrated similar clinical symptoms, which were shown in children with measles. In laboratory findings, mean WBC count was 4,933/micro L and 18 (36%) patients had thrombocytopenia. 21 (42 %) patients had complication of measles, and hepatitis was the most common episodes among them. Measles vaccination history revealed that 5 (10%) had two-dose and 7 (14%) had one-dose measled vaccine. In 35 patients, measles IgM antibody test was done. It appears that IgM positive rate among one-dose measle vaccination group, two-dose measle vaccination group, and group with unknown vaccination history were 86% (6/ 7), 40% (2/5), and 78% (18/23), respectively. Most (94%) of patients were hospitalized and more than two-third of patients were isolated. All the patients were recovered without sequelae. CONCLUSION: In 2000 measles outbreak, adult measles had the typical clinical features similar to those in the children and the severe complication was rare. Adult measles patients demonstrated low measles vaccination rate and occurred mainly among the students and office workers.
Adolescent ; Adult* ; Child ; Disease Outbreaks ; Epidemiology ; Hepatitis ; Humans ; Immunoglobulin M ; Infant ; Korea ; Male ; Measles virus ; Measles* ; Medical Records ; Occupations ; Retrospective Studies ; Thrombocytopenia ; Vaccination ; Vaccines

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Risk Factors for Prognosis in Nosocomial Acinetobacter Bacteremia.

Seung Soo SHEEN ; Young Hwa CHOI ; Yoon Jung OH ; Keu Sung LEE ; Byoung Kook IM ; Kwang Joo PARK ; Sung Chul HWANG ; Yi Hyeoung LEE

Korean Journal of Infectious Diseases.2001;33(6):436-442.

BACKGROUND: Acinetobacter bacteremia is an emerging nosocomial infection. We tried to find significant risk factors associated with the prognosis of patients with Acinetobacter bacteremia. METHODS: Retrospective case-control study was designed. The odds ratio estimation and multiple logistic regression for the categorical variables and Mann-Whitney test for the continuous variables were done. RESULTS: From September 1, 1999 to December 31, 2000 there were 25 adult patients with Acinetobacter bacteremia in Ajou University Hospital and 24 patients were confirmed as hospital acquired. The median age and hospital length of stay before bacteremia was 52 years old and 9.5 days respectively. There were 16 male patients. The overall mortality was 45.8 % (11 of 24). Thus there were 11 cases (death) and 13 controls (survival) of mortality. Statistical analysis revealed statistically significant differences between cases and controls in the terms of types of wards, central venous catheter, mechanical ventilation, total parenteral nutrition, and multi-resistant organisms. The multiple logistic regression analysis revealed that the more significant independent factors associated with mortality were mechanical ventilation and multi-resistant organisms. CONCLUSION: Acinetobacter bacteremia is a significant nosocomial infection. Especially mechanical ventilation and multi-resistant organisms were independent risk factors associated with high mortality with Acinetobacter bacteremia.
Acinetobacter* ; Adult ; Bacteremia* ; Case-Control Studies ; Central Venous Catheters ; Cross Infection ; Humans ; Length of Stay ; Logistic Models ; Male ; Middle Aged ; Mortality ; Odds Ratio ; Parenteral Nutrition, Total ; Prognosis* ; Respiration, Artificial ; Retrospective Studies ; Risk Factors*

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Characterization of Adenoviruses Isolated from Children with Respiratory Illness in Busan, 1999~2000.

Kyung Soon CHO ; Young Hee KIM ; Byoung Kuk NA ; Joo Yeon LEE ; Chun KANG ; Jae Hun BIN ; Woo Joo KIM

Korean Journal of Infectious Diseases.2001;33(6):430-435.

BACKGROUND: Adenoviruses (Ads) are a worldwide cause of endemic and epidemic respiratory infections, particularly in children, young adults and immunocompromised patients. They are responsible for 5% of the acute respiratory infections in children under the age of 4 years and account for about 10% of all cases of childhood requiring hospitalization in this age group. In this study, we report the characteristics of adenoviruses isolated from children with respiratory illness in Busan, 1999~2000. METHODS: A total of 765 children with acute respiratory illness from ten local clinics were studied. Isolation of Ads was performed by inoculating throat swab from patients into Hep-2 cells. The virus propagation was confirmed by the presence of cytopathic effect and adenoviruis specific PCR. Typing of isolated viruses was determined by sequencing analysis of hexon gene. RESULTS: Ads were isolated from throat swab of 17 (2.2%) out of 765 children. The epidemic of Ads infection was concentrated on winter months (December, January, and February). Seventeen Ad isolates showed four serotypes, with Ad3 being the most frequent strain. The distribution of Ads serotypes were eleven (64.7%) Ad3 strains, three (17.6%) Ad5 strains, two (11.8%) Ad2 srtains, and one (5.9%) Ad1 strain. CONCLUSION: Acute respiratory infections by adenoviruses in children were occurred particularly in winter season in Busan. More extended and systematic surveillance of adenovirus infection among populations is required to elucidate the extent of epidemic and disease burden of adenovirus infection.
Adenoviridae Infections ; Adenoviridae* ; Busan* ; Child* ; Hospitalization ; Humans ; Immunocompromised Host ; Pharynx ; Polymerase Chain Reaction ; Respiratory Tract Infections ; Seasons ; Young Adult

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Characteristics of Respiratory Tract Infection in the Hematopoietic Stem Cell Transplantation Population.

Dong Gun LEE ; Sang Tae PARK ; Byoung Kuk NA ; Jung Hyun CHOI ; Wan Shik SHIN ; Soon Young PAIK ; Ju Mi SHIN ; Chun KANG ; Woo Joo KIM ; Hoan Jong LEE ; Chun Choo KIM

Korean Journal of Infectious Diseases.2001;33(6):419-429.

PURPOSES: The respiratory tract infection is one of the most prevalent and serious complications following hematopoietic stem cell transplantation (HSCT). Reports not only for the respiratory tract infection but, unlikely for bacteria or fungi, for the infections caused by the respiratory viruses have been rarely reported in Korea. During the winter of 2000~2001, authors wanted to know the prevalence rate of the respiratory tract infection and the kinds of causative microorganisms, especially the community respiratory viruses (CRV). Based on these data, we attempted to evaluate the clinical courses and prognosis of the patients. METHODS: From October 2000 to February 2001, specimens were collected from the patients who visited Catholic hemopoietic stem cell transplantation center, showing symptoms and signs of respiratory tract infection after HSCT. Standard methods have been applied to isolate and identify bacterial and fungal species. Measles was diagnosed based on the typical symptoms, rash, fever, and Koplik spot. For the four different CRV (adenovirus, RSV, influenza virus, parainfluenza virus), multiplex PCR and conventional culture method were used for the identification. RESULTS: Eighty-four specimens were collected from 66 patients for 4 month period. Average age of patients was 35+/-8 years. Sixty patients (90%) were received allogeneic HSCT. Sample collection was performed between 10 and 3,740 days (average 370 days, median 215 days) after HSCT. Forty-seven patients (71.2%) have been received immunosuppressants at the time of respiratory tract infection. Forty patients (60.6 %) were suffered lower respiratory tract infection and forty-four patients (66.7%) had community-acquired infection. Sixty microorganisms were identified from 45 patients out of total 66 patients. Identified microoganisms were bacteria accounting for 2 cases (3.4%), fungi for 11 (18.3%), tuberculosis for 5 (8.3%), and viruses for 42 (70.0%). Among viruses, 16 cases were measles (39%), 14 adenovirus (33%), 9 cytomegalovirus (21%), 2 parainfluenza virus (5%), 1 was influenza virus (2%). However, no RSV was identified. Most of patients showed good prognosis without any complications. Ten (15.2%) out of total 66 patients were expired. The direct cause of death for all 8 among 10 patients was pneumonia. CONCLUSION: Of the respiratory tract infection fol-lowing HSCT, most common causative microorganisms were viruses - measles, adenovirus in order. No case of RSV infection was found. No epidemic must be occurred by influenza virus because only 1 case was found. Fourteen patients were infected by more than one microorganisms. Overall mortality rate was 15.2%. This study is still undergoing and once accumulated data for more than 1 year, it might be possible to work out a strategies of treatment and prevention for respiratory tract infections. We also expect that these data might be able to provide the basis of efficient infection control in HSCT unit.
Adenoviridae ; Bacteria ; Bone Marrow Transplantation ; Cause of Death ; Community-Acquired Infections ; Cytomegalovirus ; Exanthema ; Fever ; Fungi ; Hematopoietic Stem Cell Transplantation* ; Hematopoietic Stem Cells* ; Humans ; Immunosuppressive Agents ; Infection Control ; Korea ; Measles ; Mortality ; Multiplex Polymerase Chain Reaction ; Orthomyxoviridae ; Paramyxoviridae Infections ; Pneumonia ; Prevalence ; Prognosis ; Respiratory System* ; Respiratory Tract Infections* ; Stem Cell Transplantation ; Tuberculosis

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Iron-uptake System of Staphylococcus aureus According to the Antibiotic Susceptibility.

Sung Heui SHIN ; Yong LIM ; Nam Woong YANG ; Seung Il LEE ; Ho Jong JEON ; Jin Ho KIM

Korean Journal of Infectious Diseases.2001;33(6):414-418.

OBJECTIVE: It was reported that a few antimicrobial agents influenced on the activity of bacterial iron- uptake system (IUS). In the present study, we tried to investigate the relatedness between the resistance of antibiotics and the activity of the two high-affinity IUS, siderophore-mediated IUS and transferrin-binding protein (SA-tbp)-mediated IUS, of Staphylococcus aureus clinical isolates. METHODS: Total 71 strains including the standard ATCC 6538 strain were used. Seventy strains were isolated from the second or third line general hospitals from 1998 to 1999. Antimicrobial susceptibility test was performed by disk diffusion method. CAS agar diffusion assay was used for the measurement of staphylococcal siderophore. To visualize the expression of SA-tbp, Western blotting using human transferrin conjugated with horseradish peroxidase was performed. RESULTS: Among the nine antimicrobial agents, only the susceptible strains to oxacillin produced more siderophore compared to the resistant strains (P<0.05). There was no statistically significant difference between the antimicrobial susceptibility of the other antimicrobial agents and the production of siderophore (P>0.05). There were no antibiotics related to the expression of SA-tbp (P>0.05). CONCLUSION: These results indicate that only oxacillin (OXAC) influences on the production of staphylococcal siderophore and the further consecutive study about the action mechanism of OXAC is necessary.
Agar ; Anti-Bacterial Agents ; Anti-Infective Agents ; Blotting, Western ; Diffusion ; Horseradish Peroxidase ; Hospitals, General ; Humans ; Oxacillin ; Staphylococcus aureus* ; Staphylococcus* ; Transferrin

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Antibiogram Patterns of Staphylococcus aureus Isolates and Molecular Typing of MRSA Isolates from an Hospital in Andong City Using the Coagulase PCR-RFLP Method.

Shin KIM ; Sang Yun KIM ; Yong Ku WOO ; Eun Hee KIM

Korean Journal of Infectious Diseases.2001;33(6):404-413.

BACKGROUND: Up to date the antimicrobial resistance of Staphylococcus aureus (S. aureus) have been increased. The antimicrobial resistance patterns of Korean S. aureus strains were varied according to their isolated regions, periods, and treated method, etc. Recently methicillin-resistant S. aureus (MRSA) has already appeared one of the important nosocomial pathogens and also established the endemicity among many large scale hospitals in Korea. So far the animicrobial resistance of S. aureus isolates and MRSA isolation rates in the Andong city have been not reported yet. Therefore, it is necessary to know how many MRSA exist in one of the hospitals in Andong city. The aim of this study is to investigate the antibiogram patterns of S. aureus isolates and to differentiate the genetic relatedness among the MRSA strains using the coagulase gene product PCR-RFLP method from an hospital in Andong city over 2 years period from 1999 to 2000. METHODS: S. aureus isolates from various clinical specimens were collected from one of the hospitals in Andong city during 2 years period from 1999 to 2000. The identification of S. aureus was carried out according to the biochemical and polymerase chain reaction (PCR) method. The antimicrobial susceptibility test was carried out according to the method of NCCLLS. Methicillin resistance was tested by both MIC (microdilution method) method of NCCLLS and mecA gene PCR. The molecular typing of MRSA strains was performed by coagulase PCR-Restriction Fragment Length Polymorphism (PCR-RFLP) using the restriction enzyme (Alu I). RESULTS: One hundred fifteen S. aureus isolates were cultured from clinical specimens. Antimicrobial susceptibility test and mecA gene PCR showed that 61.4% of S. aureus isolates were identified as MRSA strains. All MRSA isolates had more than 5 drug resistance patterns. But only 34.1% (15/44) of methicillin-susceptible S. aureus (MSSA) strains had established resistence to more than 4 drugs. Depending on the results of coagulase PCR-RFLP for MRSA strains, a total of 5 genetic types were discriminated with according to the DNA band size and number. Type A and B were generally the major genetic types, which were accounted for 71.4% (50 strains) and 15.7% (11 strains), respectively and other types were isolated only in small numbers. CONCLUSIONS: MRSA strain has already been prevalent in an hospital in Andong city because 61.4% of S. aureus (70/114) isolates were typed as a MRSA strain. As the results of coagulase PCR-RFLP, type A and B were determined as a most predominant clones in this area. It is presumed that they have already been isolated from clinical specimens continuously and also established the endemicity in this area hospital. According to the diseases prevention and control strat-egy, it is necessary to block the spread of dominant MRSA clones and to study the transmission routes of dominant MRSA clones. The coagulse PCR-RFLP technique was an useful molecular typing tool for the MRSA isolates with no difficulty in manipulation.
Clone Cells ; Coagulase* ; DNA ; Drug Resistance ; Gyeongsangbuk-do* ; Korea ; Methicillin Resistance ; Methicillin-Resistant Staphylococcus aureus* ; Microbial Sensitivity Tests* ; Molecular Typing* ; Polymerase Chain Reaction ; Staphylococcus aureus* ; Staphylococcus*

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Molecular Typing of Urinary Isolates of Candida tropicalis by Pulsed-Field Gel Electrophoresis (PFGE) and Random Amplified Polymorphic DNA (RAPD) Analysis.

Joon RHO ; Chul Sung KIM ; Sook Jin JANG ; Jong Hee SHIN ; Soon Pal SUH ; Dong Wook RYANG

Korean Journal of Infectious Diseases.2001;33(6):392-403.

BACKGROUND: Despite the recognized increase of frequency of candiduria due to Candida tropicalis, little was known of its molecular epidemiology. We applied PFGE and RAPD assay for urinary C. tropicalis isolates and evaluated the utilities of PFGE and RAPD for the epidemiological typing of C. tropicalis isolates. METHODS: A total of urinary 57 isolates of C. tropicalis from 40 patients at two hospitals was analyzed. PFGE analysis were performed by electrophoretic karyotyping (EK) and restriction endonuclease analysis of genomic DNA (REAG) using two restriction enzymes (BssHII and SfiI). For RAPD, a total of 31 primers (30 random 10-mer primers and M13 primer) were used. RESULTS: EK and RAPD analysis showed the same or similar patterns among the isolates. REAG with BssHII separated 57 isolates into 28 distinct types. Six patterns were generated by REAG with using SfiI. By combining the two REAG, a total of 31 different DNA types were identified among 57 isolates from 40 patients. Three strain types were common to 23 isolates from 12 patients of a University Hospital, which suggested possible nosocomial transmission. In 19 patients with serial urinary isolates, the sequential strains from each patient exhibited the same REAG pattern. CONCLUSION: These suggest that REAG with BssHII and SfiI is useful for the investigation of molecular epidemiology of C. tropicalis isolates. In addition, some clusters of C. tropicalis isolates with the same DNA type suggest that nosocomial transmission may occur.
Candida tropicalis* ; Candida* ; DNA Restriction Enzymes ; DNA* ; Electrophoresis, Gel, Pulsed-Field* ; Humans ; Karyotyping ; Molecular Epidemiology ; Molecular Typing*

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