In spite of the extensive use of antibiotics over several decades, Streptococcus pneumoniae still remains as one of the most serious human bacterial pathogens. In order to clone the pneumococcal genes whose expression is induced when pneumococcus causes infection in mice, S. pneumoniae strain ATCC 6303 was subcultured on blood agar plates (BAP) ten times to reduce the virulence first, and then passaged through BALB/c mice three times to restore the virulence. Subtractive hybridization was performed using the total RNA preparations isolated from BAP-cultured and mouse-passaged strains. Complementary DNAs corresponding to any mRNA species that were differentially expressed in the mouse- passaged strain were used as the probes to screen the genomic library of pneumococcus. Positive recombinants were selected and sequenced partially to identify the genes located within the cloned DNA. GenBank search of the sequence data has identified several genes including two heat shock genes (dnaK and dnap, a transposase-encoding gene, and a sequence which is very homologous to that of the ftsH gene.
Agar ; Animals ; Anti-Bacterial Agents ; Clone Cells* ; Cloning, Organism* ; Databases, Nucleic Acid ; DNA ; DNA, Complementary ; Genomic Library ; Hot Temperature ; Humans ; Mice ; Pneumonia ; RNA ; RNA, Messenger ; Shock ; Streptococcus pneumoniae* ; Streptococcus* ; Virulence

No abstract available.

BACKGROUND: Identification of specific antinuclear antibodies is useful for the diagnosis, subclassification and determination of prognosis in autoimmune disorders. In many diseases, multiple autoantibodies are detected, and simultaneous detection of multiple autoantibodies has been shown to be useful. Recently, a commercial kit (EL-ANA/6 profiles, TheraTest Laboratories, USA) losing enzyme-linked immunosorbent assay (ELISA) method for detection of six specific autoantibodies is avallable. In this study, we attempted to compare the results of EL-ANA/6 profiles with those of routinely used methods and evaluated usefulness of EL-ANA/6 profiles. METHODS: EL-ANA/6 profiles were performed with 28 sera which were positive for fluorescent antinuclear antibody (FANA) Simultaneously we tested anti-dsDNA antibodies with immnofluorescent (If) method and anti-Sm, anti-RNP, anti-SSA and anti-SSB antibodies with double immunodiffusion (DID). To evaluate specificity, EL- ANA/6 profiles tests were performed on 10 sera from healthy blood donors. RESULTS: Ten sera of healthly blood donors were all negative for EL-ANA/6 pro biles. In the results of EL-ANA/6 profiles on sera positive for FANA, the concordance rate with IF method for the anti-dsDNA antibodies was 89.3% (25/28) and the con- cordance rates with DID for anti-Sm, anti-Sm/RNP, anti-SSA and anti-SSB antibodies were 85.7% (24/28), 82.1% (23/28), 92.9% (26/28) and 82.1% (23/28), respectively. In 16 discordant settings, thirteen (81.3%) were negative on DID and positive on EL-ANA/6 profiles. CONCLUSIONS: The results of the EL-ANA/6 profiles show good concordance rates with If and DID. EL-ANA/6 profiles showing quantitative profiles for multiple autoantibodies is useful for diagnosis and tool)ow-up of autoimmune disorders.
Antibodies ; Antibodies, Antinuclear* ; Autoantibodies ; Bile ; Blood Donors ; Diagnosis ; Enzyme-Linked Immunosorbent Assay ; Humans ; Immunodiffusion ; Prognosis ; Sensitivity and Specificity

No abstract available.
Arthritis* ; Knee*

BACKGROUND: Methicillin-resistant Staphylococcus aureus(MRSA) is an increasingly common cause of nosocomial infections worldwide. Epidemiologic investigation of MRSA outbreaks and identification of pathways of nosocomial MRSA spread require the ability to distinguish individual MRSA strains. We applied molecular tap ing of the methicillin-resistant determinant (mec) and coagulase typing in the investigation of a nosocomial MRSA infections. METHODS: We randomly selected 79 strains of mecA positive MRSA isolated from patients who visited Dong-A university Hospital from Dec. 1995 to Oct. 1996. Molecular typing of MRSA was performed by comparing the size of the mac-associated hypervariable region amplified by the polymerase chain reaction (PCR). Coagulase typing with type I-VIII antisera was also used for classification of MRSA based on its phenotype. Each isolates were classified by the combination of molecular analyses and coagulase type. RESULTS: The 79 MRSA isolates were grouped Into sin hypervariable legion (HVR) genotypes on the basis of the size of the PGR products. In coagulase typing, the most predominant type was II(46.8%) and type V was not found. Nine strains were not typable. The combination of HVR genotypes and coagulase types showed 23 different types in 79 MRSA Isolates. The strains which were repeatedly isolated from the same patients showed the same HYR genotypes and coagulate types. CONCLUSION: The combination of HVR genotypes and coagulase types is thought to be useful in epidemiolgical Investigation of nosocomial infections caused by MRSA ,because of its simplicity and reproducibility.
Classification ; Coagulase* ; Cross Infection* ; Disease Outbreaks ; Genotype ; Humans ; Immune Sera ; Methicillin Resistance* ; Methicillin-Resistant Staphylococcus aureus* ; Molecular Typing* ; Phenotype ; Polymerase Chain Reaction ; Staphylococcus

BACKGROUND: Recent DNA polymorphism analysis using numerous DNA markers has been used to determine the parental origin of the extra chromosome 21 in Down syndrome. In this study we used seven dinucleotide repeat polymorphisms on chromosome 21 to characterize a case of rea(21q21q) and to know whether it is consistent with an isochromosome or a true Robertsonian translocation. METHODS: Cytogenetic investigation was done by conventional G banding DNA was extracted from whole blood of a proband and her parents and was amplified by PCR using seven sets of (GT)n repeat dinucleotide markers located on the long arm of chromosome 21 After electrophoresis of the PCR product in polyacrylamide gel and silver staining the parental origin and number of DNA copy were determined by visual comparison of the band intensities within and between individuals. RESULTS: Conventional cytogenetics showed that the proband had a 46.XX.re(21q21q) chromosome pattern. Parental chromosome studies were normal, therefore, the rearrangement was a de novo event. All seven DNA markers showed one or two alleles, demonstrating rea(21q21q) to be an isochromosome. For D21S215 and D21S156 markers both parents were heterozygous and the proband inherited one copy of paternal allele and two copies of maternal allele which both parents did not share. This finding was consistent with a maternally derided isochromosome. CONCLUSION: Use of dinucleotide repeat DNA polymorphisms after PCR amplification will be very useful to detect the parental origin of additional chromosome 21 or rearrangement of chromosome 21 in Down syndrome. Besides employing siltier staining of a PCR product we will be able to avoid using of radioisotopes and apply to clinical laboratory diagnosis.
Alleles ; Arm ; Chromosomes, Human, Pair 21 ; Clinical Laboratory Techniques ; Cytogenetics ; Dinucleotide Repeats* ; DNA ; Down Syndrome ; Electrophoresis ; Genetic Markers ; Humans ; Isochromosomes ; Parents ; Polymerase Chain Reaction ; Radioisotopes ; Silver Staining

Classification of meniscal tear is necessary only for partial resection. In western countries many reports said that medial meniscal tear is commoner than that of lateral meniscus. Actually the reverse is true in Korea and in some other oriental countries, However there is no plausible explanation concerning this matter. In this report 250 knees of 240 patients were involved. We performed arthroscopic meniscectomy in all cases and we investigated the relationship between the pattern of tear and many possibly related factors. The follow-up period was 1–8 years, the ratio of male-female was 163:77, and the range of age was varied between 7 and 71 tears with the young and middle age groups being predominant. Right and left side ratio was 117:113 and the involvement of both knees was noted in 20 knees of 10 patients. The results were as follows: 1. The medial menisci were torn in 100 knees and was less than 142 lateral meniscal tear, with the ratio being 1:1.4. The tear of lateral meniscus was commoner in the age group of less than 30 years and the tear of medial meniscus was commoner in the age group older than 50 years whereas there was no difference in the age group of inbetween. 2. The commonest pattern of tear was longitudinal one including peripheral tear, that is 135 knees (54.0%), followed by complex tear, 50 knees (20.0%), oblique tear, 40 knees (16.0%), horizontal tear, 21 knees (8.4%) and lastly superior and inferior flap tear, 5 knees (2.0%). 3. The longitudinal tear was most prevalent in the age group of 20–40, whereas the complex tear was most prevalent in the age group of beyond 50 years (p<0.05). 4. The longitudinal tear was significantly prevalent in lateral menisci, the complex tear in medial menisci (p<0.05), whereas the transverse and oblique tear were not significantly prevalent in either meniscus (p>0.05). 5. In the longitudinal tear group, the tear occured most frequently in the midsubstance of posterior portion in the medial menisci without statistical significance (p>0.05) wereas the tear occurred most frequently in the periphery of the anterior portion in case of lateral menisci (p<0.05). 6. There were 37 discoid lateral menisci, with the longitudinal tear being the commonest. 7. Ground sports injuries occurred only in 34 knees (13.6%), with the longitudinal tear being significantly commoner than in the other injuries. From these results it would be suggested that although the discoid lateral meniscus has to be given much weight in the total meniscal injuries another explanation should be given concerning the prevalence of lateral meniscus tear compared to the medial meniscus tear and the prevalence of longitudinal tear.
Athletic Injuries ; Classification ; Follow-Up Studies ; Humans ; Knee ; Korea ; Menisci, Tibial ; Middle Aged ; Prevalence ; Tears

No abstract available.
Respiratory Syncytial Virus Infections* ; Respiratory Syncytial Viruses*

The gene ftsH encodes a membrane-bound and ATP-dependent protease that is involved in a variety of cellular functions including heat-shock and stress response. Streptococcus pneumoniae DNA encompassing most part of the ftsH gene was cloned in Escherichia coli and sequenced. Due to the unsuccessful cloning as seen in other pneumococcal promoters, the 5'-end of the gene including the upstream promoter region was amplified by inverse polymerase chain reaction and then sequenced by cyclic sequencing. The amino acid sequence that is deduced from the 1,959 bp-long ftsH gene is very similar to FtsH of several gram-positive bacteria and E. coli within the region responsible for the AAA (ATPase associated with diverse cellular activities) function. Except for the N-terminal domain that contains a short extracellular region between two mernbrane-spanning segments, pneumococcal FtsH shows striking sequence similarity to that of a closely related species Lactococcus lactis within the conserved cytoplasmic domain where two ATP-binding motifs, the AAA Signature motif, and a zinc-binding motif are found.
Amino Acid Sequence ; ATP-Dependent Proteases ; Base Sequence* ; Clone Cells* ; Cloning, Organism* ; Cytoplasm ; DNA ; Escherichia coli ; Gram-Positive Bacteria ; Lactococcus lactis ; Polymerase Chain Reaction ; Promoter Regions, Genetic ; Streptococcus pneumoniae* ; Streptococcus* ; Strikes, Employee

We examined the expressions of CD44 splice variants (v4/5, v6), alpha-smooth muscle actin, nm23 to evaluate their roles as prognostic factors in 70 cases of uterine cervical carcinoma (stage IB to IIB) who were surgically treated from January 1989 to June 1990 with a clinical follow-up of a minimum of 5 years. The expression was examined by an immunohistochemical method using archival formalin fixed paraffin embedded tissue. In the 70 cases, 61 cases were squamous cell carcinoma and 9 cases were adenocarcinoma. CD44v4/5, CD44v6, alpha-smooth muscle actin, and nm23 were detected in 41.4%, 70%, 100%, and 74.3% of tumor samples, respectively. CD44 splice variants and nm23 showed membrane and cytoplasmic staining of tumor cells, respectively. The expression of alpha-smooth muscle actin showed cytoplasmic staining confined to stromal cells and was classified into three grades by the extent in stromal cells: with less than 10% of stromal cells; 32.9%, 10-50% of stromal cells; 40.0%, more than 50%; 27.1%. These expressions were not correlated with histologic types, lymph node involvement, recurrence, and grades of tumor infiltrating lymphocyte (TIL). But CD44v4/5 had significantly inverse correlation with TIL (p=0.049). The expression of CD44v4/5 was significantly correlated with that of CD44v6 (p=0.05), and that of alpha-smooth muscle actin was inversely correlated with that of nm23 (p=0.049). In conclusion, in FIGO IB-IIB uterine cervical carcinoma CD44 variants, nm23, and SMA show high prevalence, however, with little prognostic significance assessed by recurrence and lymph node metastasis.
Actins* ; Adenocarcinoma ; Carcinoma, Squamous Cell ; Cytoplasm ; Follow-Up Studies ; Formaldehyde ; Lymph Nodes ; Lymphocytes ; Membranes ; Neoplasm Metastasis ; Paraffin ; Prevalence ; Recurrence ; Stromal Cells ; Uterine Cervical Neoplasms*