BACKGROUND: The lineage assignment in acute leukemias is critical in therapeutic decisions. Immunophenotyping by flow cytometry plays the main role in the lineage assignment; however, few studies have been done to determine the optimal set of markers. In this regard, we tried to find out the optimal first-line set of markers with a minimal compromise in its diagnostic sensitivity. MATERIALS AND METHODS: We retrospectively analyzed 321 cases of acute leukemias whose diagnoses were based on the EGIL (European Group for Immunological Classification of Acute Leukemia) scores. At our institution, flow cytometic analyses included 15 first-line markers and 4 additional second-line markers as needed, along with immunohistochemical stains. We performed simulational studies for the expected EGIL scores involving every possible combination of markers and analyzing the overall diagnostic sensitivities in each combination. RESULTS: The cytoplasmic antigens including MPO stain and CD79a stain contributed greatly to the lineage assignment. For a sensitivity over 95%, there needed a combination of MPO stain with other 5 flow markers (CD33, CD13, CD14, CD15 and CD117) for myeloid lineage; CD79a stain with 3 flow markers [CD19, CD10, and CD20 (or TdT)] for B-lymphoid lineage; and 4 flow markers (CD2, CD3, CD5, and CD7) for T-lymphoid lineage. CONCLUSIONS: To maintain diagnostic sensitivities over 95% for each lineage, at least 14 markers (including MPO stain and CD79a stain) were needed; while 16 markers were needed for a sensitivity of 100%. When combined with other important markers for specific aims such as CD45, the minimum number of markers needed for the accurate diagnosis of acute leukemias would be more than about 18 to 20.
Classification ; Coloring Agents ; Cytoplasm ; Diagnosis ; Flow Cytometry ; Immunophenotyping ; Leukemia* ; Retrospective Studies

BACKGROUND: The CD34+ cell dose and infused number of committed progenitor cells in transplantation are important factors in hematologic engraftment. However, the relationship between expansion potential of progenitor cells and hematologic engraftment remains controversial. We evaluated whether expansion potential of progenitor cells is a predictive factor of post-transplantation hematologic engraftment. METHODS: Mononuclear cells isolated from mobilized peripheral blood and bone marrow were cultured with cytokine cocktail for 7 days. Progenitor cells and committed progenitors were analyzed using stem cell markers (CD34 and CD133) and lineage specific markers. Hematologic engraftment was defined as neutrophil counts over 500/microliter and platelet counts over 20,000/microliter without transfusion. Acute and chronic graft-versus-host disease (GVHD) were investigated. RESULTS: There was inverse tendency between the number and fold expansion of progenitor cells or committed (granulocytic or megakaryocytic) progenitors and time to engraftment. Especially, fold expansion of CD34(+)/CD33(+) cells was significantly correlated with time to neutrophil engraftment in bone marrow transplantation (r=-0.56, P=0.04). The infused number and fold expansion of lymphoid progenitors were not related to the occurrence of acute or chronic GVHD. CONCLUSIONS: We could not prove that expansion potential of progenitor cells and committed progenitor cells is correlated to hematologic engraftment although there is a correlation between CD34(+)/ CD33(+) cells and time to neutrophil engraftment. But, a further study on the value of expansion potential is required because there is an inverse tendency.
Bone Marrow ; Bone Marrow Transplantation ; Graft vs Host Disease ; Hematopoietic Stem Cell Transplantation* ; Hematopoietic Stem Cells* ; Neutrophils ; Platelet Count ; Stem Cells

BACKGROUND: Cystic fibrosis (CF) is one of the most common hereditary disorders among Caucasians. The most common mutations of the cystic fibrosis transmembrane conductance regulator (CFTR) gene have been well established among Caucasian populations. In Koreans, however, there are very few cases of genetically confirmed CF thus far, and the spectrum of mutations seems quite different from that observed in Caucasians. METHODS: In the present study, we describe the cases of 2 Korean CF patients, present sequencing results identifying mutations in their CFTR gene, and summarize the results of CFTR mutational spectrum from previously reported Korean CF patients. The mutations described were identified by performing direct sequencing analysis of the complete coding regions and flanking intronic sequences of the CFTR gene, followed by multiplex ligation-dependent probe amplification (MLPA) analysis in order to detect gene deletions or duplications that could not be identified by a direct sequencing method. RESULTS: Three CFTR mutations were identified in the 2 patients, including p.Q98R, c.2052delA, and c.579+5G>A. In an analysis of 9 Korean CF patients that included the 2 patients presented in this study, p.Q98R mutation was the only recurrently observed mutation with a frequency of 18.8% (3/16 alleles). Furthermore, only one of the mutations (c.3272-26A>G) was found among the 32 common mutations in the screening panel for Caucasians from the Cystic Fibrosis Mutation Database. CONCLUSIONS: Sequencing of the entire CFTR gene followed by MLPA analysis, rather than using the targeted sequencing-based screening panel for mutations commonly found in Caucasian populations, is recommended for genetic analysis of Korean CF patients.
Adult ; Alleles ; Asian Continental Ancestry Group/*genetics ; Cystic Fibrosis/*genetics ; Cystic Fibrosis Transmembrane Conductance Regulator/*genetics ; Female ; Heterozygote ; Humans ; Male ; Mutation ; Republic of Korea ; Sequence Analysis, DNA ; Young Adult

BACKGROUND: The objective of this study was to explore whether individual variations in the concentration of growth factors (GFs) influence the biologic effects of platelet-rich plasma (PRP) on human mesenchymal stem cells (HMSCs). METHODS: The concentrations of 7 representative GFs in activated PRP (aPRP) were measured using ELISA. The effects of PRP on the proliferation and alkaline phosphatase (ALP) activity of HMSCs were examined using several concentrations of aPRP from 3 donors; the relationships between the GF levels and these biologic effects were then evaluated using 10% aPRP from 5 subgroups derived from 39 total donors. HMSCs were cultured in DMEM with the addition of aPRP for 4 or 12 days; then, DNA content and ALP activity were measured. RESULTS: The quantity of DNA increased significantly at a 10% concentration of aPRP, but the ALP activity was suppressed at this concentration of aPRP. The GF concentrations varied among donors, and 5 subgroups of characteristic GF release patterns were identified via cluster analysis. DNA levels differed significantly between groups and tended to be higher in groups with higher concentrations of transforming growth factor-beta1 (TGF-beta1) and platelet-derived growth factors (PDGFs). DNA quantity was positively correlated with TGF-beta1 concentration, and was negatively correlated with donor age. ALP activity was negatively correlated with PDGF-BB concentration. CONCLUSIONS: The varying GF concentrations may result in different biologic effects; thus, individual differences in GF levels should be considered for reliable interpretation of the biologic functions and standardized application of PRP.
Alkaline Phosphatase/metabolism ; Blood Donors ; Cell Differentiation ; Cells, Cultured ; Culture Media/chemistry ; DNA/analysis ; Humans ; Intercellular Signaling Peptides and Proteins/*pharmacology ; Mesenchymal Stem Cells/*cytology/drug effects ; Platelet-Derived Growth Factor/pharmacology ; Platelet-Rich Plasma/*metabolism ; Transforming Growth Factor beta1/pharmacology

Streptococcus suis infection is an emerging zoonosis in Asia. The most common disease manifestation is meningitis, which is often associated with hearing loss and cochleovestibular signs. S. suis infection in humans mainly occurs among risk groups that have frequent exposure to pigs or raw pork. Here, we report a case of S. suis meningitis in a 67-yr-old pig carcass handler, who presented with dizziness and sensorineural hearing loss followed by headaches. Gram-positive diplococci were isolated from cerebrospinal fluid (CSF) and blood cultures and showed gray-white colonies with alpha-hemolysis. S. suis was identified from CSF and blood cultures by using a Vitek 2 system (bioMerieux, France), API 20 STREP (bioMerieux), and performing 16S rRNA and tuf gene sequencing. Even after receiving antibiotic treatment, patients with S. suis infection frequently show complications such as hearing impairment and vestibular dysfunction. To the best of our knowledge, this is the first case of S. suis meningitis in Korea. Prevention through public health surveillance is recommended, especially for individuals who have occupational exposures to swine and raw pork.
Aged ; Animals ; Bacterial Proteins/genetics ; Blood/microbiology ; Cerebrospinal Fluid/microbiology ; Hearing Loss, Bilateral/complications/*diagnosis/microbiology ; Humans ; Male ; Meningitis, Bacterial/complications/*diagnosis/microbiology ; Phylogeny ; RNA, Ribosomal, 16S/genetics ; Sequence Analysis, DNA ; Streptococcus suis/classification/genetics/*isolation & purification ; Swine ; Tomography, X-Ray Computed

Phaeohyphomycosis is a subcutaneous infection caused by dark pigmented fungi, including fungi of the species Phaeoacremonium, Alternaria, Exophiala, and Pyrenochaeta. In August 2005, a 54-yr-old man who had received a renal transplant 5 yr ago was admitted to our hospital with a subcutaneous mass on the third finger of the right hand; the mass had been present for several months. He had been receiving immunosuppressive agents for several years. He underwent excision of the mass, which was followed by aspiration of the wound for bacterial and fungal cultures. Many fungal hyphae were observed on the histology slide treated with periodic acid-Schiff stain. A few white waxy colonies with a woolly texture grew on the Sabouraud dextrose agar at 30degrees C and changed to dark brown in color. Nucleotide sequencing of internal transcribed spacer regions revealed 100% homology to the Phaeoacremonium aleophilum anamorph and Togninia minima teleomorph (514 bp/514 bp). The patient completely recovered after wide surgical excision. Here, we report the first case of phaeohyphomycosis caused by Phaeoacremonium species in a kidney transplant patient in Korea.
Antifungal Agents/therapeutic use ; Ascomycota/genetics/*isolation & purification ; Dermatomycoses/drug therapy/etiology/*microbiology ; Fingers/surgery ; Humans ; Immunosuppressive Agents/adverse effects ; *Kidney Transplantation ; Male ; Middle Aged ; Republic of Korea ; Sequence Analysis, DNA ; Subcutaneous Tissue/microbiology

Bacillus Calmette-Guerin (BCG) has been traditionally used as a vaccine against tuberculosis. Further, intravesical administration of BCG has been shown to be effective in treating bladder cancer. Although BCG contains a live attenuated strain of Mycobacterium bovis, complications such as M. bovis BCG infection caused by BCG administration are extremely rare. Here, we report a case of BCG infection occurring after intravesical BCG therapy. A 67-yr-old man presented with azotemia and weight loss. He had been diagnosed with bladder cancer 4 yr back, and had undergone transurethral resection of the bladder tumor and intravesical BCG (Tice strain) therapy at that time. An acid-fast bacterial strain was isolated from his urine sample. We did not detect Mycobacterium tuberculosis protein 64 (MPT-64) antigen in the isolates obtained from his sample, and multiplex PCR and PCR-reverse blot hybridization assay indicated that the isolate was a member of the M. tuberculosis complex, but was not M. tuberculosis. Finally, sequence analysis of 16S ribosomal RNA and DNA gyrase, subunit B (gyrB) suggested that the organism was M. bovis or M. bovis BCG. Although we could not confirm that M. bovis BCG was the causative agent, the results of the 3 molecular methods and the MPT-64 antigen assay suggest this finding. This is an important finding, especially because M. bovis BCG cannot be identified using common commercial molecular genetics tools.
Administration, Intravesical ; Aged ; BCG Vaccine/administration & dosage/*adverse effects ; DNA Gyrase/genetics ; Humans ; Male ; Mycobacterium Infections/*diagnosis/etiology ; Mycobacterium bovis/genetics/*isolation & purification ; Polymerase Chain Reaction ; RNA, Ribosomal, 16S/genetics ; Urinary Bladder Neoplasms/*therapy

BACKGROUND: Clostridium difficile infection (CDI) has markedly risen and is associated with hypervirulent ribotype 027 outbreaks in North America and Europe since 2003. The aims of this study were to determine the prevalence of ribotype 027 among C. difficile isolates in Korea, to characterize the ribotype 027 isolates, and to determine the clinical severity of CDI in patients infected with these isolates. METHODS: A total of 1,251 isolates of C. difficile recovered from stool specimens of suspected CDI patients at two tertiary-care hospitals and one commercial laboratory between 2002 and 2009. Genes for toxin A (tcdA), toxin B (tcdB), and binary toxin (cdtA and cdtB) were detected by PCR. Mutation in the tcdC gene was detected by sequencing after PCR amplification. For molecular genotyping, we performed PCR-ribotyping, pulsed-field gel electrophoresis (PFGE), and multilocus variable-number tandem-repeat analysis (MLVA). Minimum inhibitory concentrations of moxifloxacin were determined using Etest strips (AB bioMerieux, Sweden). RESULTS: We identified 7 isolates as ribotype 027. These isolates had the same tcdC mutation as the epidemic strain, and 6 of them were resistant to moxifloxacin. The isolates were categorized into 3 different PFGE types and 7 different MLVA types. All the 7 cases had occurred sporadically. CONCLUSIONS: C. difficile ribotype 027 is uncommon, but it has emerged in Korea. The spread of this ribotype should be closely monitored in order to avoid an outbreak of CDI in Korea.
Adult ; Aged ; Aged, 80 and over ; Bacterial Proteins/genetics/metabolism ; Bacterial Toxins/genetics/metabolism ; Clostridium difficile/genetics/*isolation & purification ; Drug Resistance, Bacterial ; Electrophoresis, Gel, Pulsed-Field ; Enterocolitis, Pseudomembranous/microbiology ; Enterotoxins/genetics/metabolism ; Feces/microbiology ; Female ; Humans ; Male ; Microbial Sensitivity Tests ; Middle Aged ; Mutation ; Polymerase Chain Reaction ; Republic of Korea ; *Ribotyping

BACKGROUND: In order to determine the clinical usefulness of the MicroScan (Siemens Healthcare Diagnostics, USA) MICroSTREP plus antimicrobial panel (MICroSTREP) for testing antimicrobial susceptibility of beta-hemolytic streptococci (BHS) and viridans group streptococci (VGS), we compared the accuracy of MICroSTREP with that of the CLSI reference method. METHODS: Seventy-five BHS and 59 VGS isolates were tested for antimicrobial susceptibility to ampicillin, penicillin, cefotaxime, meropenem, erythromycin, clindamycin, levofloxacin, and vancomycin by using MICroSTREP and the CLSI agar dilution method. RESULTS: The overall essential agreement with regard to minimum inhibitory concentrations (MICs) (within +/-1 double dilution) between MICroSTREP and the CLSI reference method was 98.2%, and categorical agreement (CA) was 96.9%. For the BHS isolates, the CA for erythromycin was 96.0%, whereas that for cefotaxime, meropenem, levofloxacin, and vancomycin (for ampicillin, penicillin, and clindamycin; 98.7%) was 100%. For the VGS isolates, the CA for penicillin was 84.7% and that for erythromycin, clindamycin, and vancomycin (for meropenem, 86.5%; for ampicillin, 88.1%; and for cefotaxime and levofloxacin, 96.6%) was 100%. All categorical errors of penicillin and ampicillin in the VGS isolates were minor. CONCLUSIONS: The accuracy of MICroSTREP is comparable to that of the CLSI reference method, suggesting that this panel can be effective for testing antimicrobial susceptibility of BHS and VGS.
Anti-Bacterial Agents/*pharmacology ; Drug Resistance, Bacterial ; Humans ; Microbial Sensitivity Tests ; Reagent Kits, Diagnostic ; Streptococcal Infections/microbiology ; Streptococcus/*drug effects/isolation & purification ; Viridans Streptococci/*drug effects/isolation & purification

BACKGROUND: The origin of hematologic malignancies has been known to be monoclonal. In most cases, the same or obviously related chromosomal abnormliaties are found and cytogenetically unrelated clones are uncommon. We evaluated the prevalence and clinical significance of patients with cytogenetically unrelated clones in hematologic malignancies. METHODS: Included in the study were 324 patients who had been diagnosed with the following hematologic malignancies at Ewha Womans University, Mokdong Hospital: AML (93 cases), MDS (27), CML (51), myeloproliferative disorder (38), acute biphenotypic leukemia (8), ALL (44), CLL (9), multiple myeloma (MM, 40), and Non-Hodgkin's lymphoma with bone marrow involvement (14). RESULTS: The overall prevalence of hematologic malignancies with cytogenetically unrelated clones at diagnosis was 0.9% (3/324). Of AML patients, 1.1% (1/93) had unrelated clones, CLL 11.1% (1/9), and MM 2.5% (1/40). The other hematologic malignancies did not show cytogenetically unrelated clones. The AML patient had add(11)(q23)/add(1)(p36.3); the CLL patient had +12/ del(13)(q22); and the MM patient had +der(1)t(1;13)(p12;q12), -13/-X, +5, +7, -8, -12, -13, add(14) (q32), +15, -16, +19, -20, -22, -22. We also detected an unrelated clone of trisomy 8 in Philadelphia chromosome negative cells from a CML patient who was treated with imatinib mesylate. CONCLUSIONS: Hematologic malignancies with cytogenetically unrelated clones are uncommon. This report highlights the importance of the conventional chromosomal analysis in that an unrelated clone in philadelphia chromosome negative cells may be detected in a CML case.
Bone Marrow ; Clone Cells* ; Diagnosis ; Female ; Hematologic Neoplasms* ; Humans ; Leukemia, Biphenotypic, Acute ; Lymphoma, Non-Hodgkin ; Mesylates ; Multiple Myeloma ; Myeloproliferative Disorders ; Philadelphia Chromosome ; Prevalence ; Trisomy ; Imatinib Mesylate