Constitutive stabilization of hypoxia-inducible factor alpha selectively promotes the self-renewal of mesenchymal progenitors and maintains mesenchymal stromal cells in an undifferentiated state

With the increasing use of culture-expanded mesenchymal stromal cells (MSCs) for cell therapies, factors that regulate the cellular characteristics of MSCs have been of major interest. Oxygen concentration has been shown to influence the functions of MSCs, as well as other normal and malignant stem cells. However, the underlying mechanisms of hypoxic responses and the precise role of hypoxia-inducible factor-1alpha (Hif-1alpha), the master regulatory protein of hypoxia, in MSCs remain unclear, due to the limited span of Hif-1alpha stabilization and the complex network of hypoxic responses. In this study, to further define the significance of Hif-1alpha in MSC function during their self-renewal and terminal differentiation, we established adult bone marrow (BM)-derived MSCs that are able to sustain high level expression of ubiquitin-resistant Hif-1alpha during such long-term biological processes. Using this model, we show that the stabilization of Hif-1alpha proteins exerts a selective influence on colony-forming mesenchymal progenitors promoting their self-renewal and proliferation, without affecting the proliferation of the MSC mass population. Moreover, Hif-1alpha stabilization in MSCs led to the induction of pluripotent genes (oct-4 and klf-4) and the inhibition of their terminal differentiation into osteogenic and adipogenic lineages. These results provide insights into the previously unrecognized roles of Hif-1alpha proteins in maintaining the primitive state of primary MSCs and on the cellular heterogeneities in hypoxic responses among MSC populations.

A Case of Immune Hemolytic Anemia Induced by Ceftizoxime and Cefobactam (Sulbactam/Cefoperazone) / 대한진단검사의학회지

Simultaneous drug-induced immune hemolytic anemia (DIIHA) caused by multiple drugs is rare. We report a case of a patient who developed DIIHA caused by 2 drugs. The patient's serum exhibited agglutination of ceftizoxime- or sulbactam-coated red blood cells (RBCs; via a drug-adsorption mechanism) and of uncoated RBCs in the presence of sulbactam (via an immune-complex mechanism). Although ceftizoxime is known to exhibit a positive reaction by an immune-complex method with or without reactivity with drug-coated RBCs, this patient's antibodies were reactive only against drug-coated RBCs. On the other hand, sulbactam, which is known to cause hemolytic anemia by nonimmunologic protein adsorption, exhibited positive reactions in tests with both drug-coated RBCs and in the presence of sulbactam. This is the first report of DIIHA due to a sulbactam-cefoperazone combination and the fourth report of DIIHA due to ceftizoxime. Owing to the patient's complicated laboratory results, DIIHA was suspected only at a late stage. We propose that for the prompt diagnosis of DIIHA, tests for all possible causative drugs should be conducted by 2 methods.

Subjective and Objective Results of Radiofrequency Ablation in Drug-Resistant Allergic Rhinitis Based on a 12-Month Follow-up Study

BACKGROUND AND OBJECTIVES: Allergic rhinitis is a common condition that has been treated using various methods. Nevertheless, it remains an intractable condition in some cases. We evaluated the long-term efficacy of radiofrequency ablation (RFA) therapy for drug-resistant allergic rhinitis. MATERIALS AND METHODS: Eighteen patients with allergic rhinitis were enrolled in this prospective interventional study. Their subjective symptoms and objective findings were assessed using a visual analogue scale and acoustic rhinometry before and after RFA therapy. Postoperative follow-up was carried out at 1, 3, 6, and 12 months. RESULTS: Nasal obstruction and rhinorrhea were improved significantly (p<0.001 and p=0.015, respectively), and these effects continued for 12 months after RFA therapy. However, the effects on nasal itching and sneezing did not last throughout the year of the study. Patients' overall satisfaction scores declined at 12 months. On acoustic rhinometry, the minimal cross-sectional area and the total nasal volume did not show improvement, and there were no correlations with the subjective symptoms. Most of the complications associated with the RFA procedure were minimal and resolved within 4 weeks. CONCLUSION: RFA is a safe, simple, and effective procedure for relieving nasal obstruction and rhinorrhea for at least 12 months in drug-resistant allergic rhinitis.

Evaluation of Single-use Blood Donor Arm Antiseptic Kits / 대한수혈학회지

BACKGROUND: Transfusion-associated bacterial contamination (TABC) have become one of the major hazards of transfusion and the donors' normal skin flora is considered to be the most important source of contamination. We therefore evaluated the effectiveness of single-use blood donor arm antiseptic kits available on the Korean market. METHODS: Forty-eight antiseptic kits of two manufacturers consisting of 70% isopropyl alcohol swab stick and 10% povidone iodine swab stick were evaluated for their performance in reducing the bacterial load of the skin before and after disinfection. Staff response about ease of use was also evaluated. RESULTS: Whereas before disinfection the mean bacterial count was 5 CFU/plate or less, after disinfection bacterial growth was hardly observed. Overall both antiseptic kits showed a percent reduction of more than 98% compared to predisinfection. The amount of antiseptic solution of one manufacturers' kit was excessive and leakage of the antiseptic solution was also observed. CONCLUSION: The effectiveness of both antiseptic kits evaluated in this study was satisfactory to be used for blood donors, but an improvement to the packaging of one manufacturers' kit is required before it can be used in the field.

Evaluation of Nested Multiplex PCR in the Diagnosis of Malaria Infection / 대한진단검사의학회지

BACKGROUND: A peripheral blood smear has been the gold standard method for the diagnosis of malaria infection. Recently, many other methods have been introduced, although having inferior sensitivity and specificity to peripheral blood smears. We evaluated Neodin malaria PCR kit and its applicability in clinical settings. METHODS: Samples from seventy patients who visited Korea University hospital were used for evaluation. DNA from EDTA blood was tested in nested multiplex PCR and 470 bp for Plasmodium vivax or 340 bp for Plasmodium falciparum was confirmed after electrophoresis. The detection limit was determined by dilution of malaria positive blood with normal blood. RESULTS: Thirty-five cases of P. vivax and 10 cases of P. falciparum were noted. Except for a case of falciparum malaria, all positive cases were consistent with the peripheral blood smear results. Detection limit was 3.6 parasite/microL. CONCLUSIONS: Neodin malaria nested multiplex PCR has high sensitivity and the ability for species discrimination and may be available in the diagnosis of malaria infection.

A Psychopathology and Socio-Psychiatric Study of Delinquent Adolescents doing School Violence / 신경정신의학

OBJECTIVE: The purpose of this study was to identify the characteristics of school violence, self concept, psychopathology and psychosocial characteristics of male aldolescent delinquency doing school violence. And we hoped to use this result as basic material for prevention of school violence through comparison of those of students group. METHOD: We surveyed 725 male adolescents including 331 deliquents from educational program called "Big Brothers and Sisters" in Pusan. We used self-reporting questionnaire composed of demographic data and attitude to violence. In order to investigate the psychopathology and self-concept, we used Child Behavior Check List, Youth edit, Korean version(CBCL) and the Piers-Harris Children's Self-Concept Scale. Statistical methods employed were Chi-square, t-test, two-way ANOVA. RESULT: The results show that in most demographic and psychosocial factor including age, education, socio-economic condition, religion and relationship with friends, there was no significant difference between delinquent and student group. And there was no significant difference between bully and non-bully in both group. The frequency of alcohol drinking and smoking were higher in bullying groups. For the feeling of watching violent mass-media, bullying delinquents reported more excited state than non-bullying delinquents in both group. On results of CBCL, subscales of delinquent behavior, aggressive behavior, externalizing problem, social scale in delinquent group were higher than those in student group. Subscale of somatic complaints, thought problem, attention problem, delinquent behavior, aggressive behavior, internalizing problem, and externalizing problem in bullying group were higher than those in nonbullying group. In both delinquent and student group, subscale of delinquent behavior, aggressive behavior, and externalizing problem in bullying group were significantly higher than those in nonbully group. In the results of self-concept scale, subscales of behavior, intellectual and school status in delinquent group were lower than those in student group. Subscale of somatic complaints, thought problem, attention problem, delinquent behavior, aggressive behavior, internalizing problem, and externalizing problem in bullying group were higher than those in nonbully group. CONCLUSION: In both delinquent and student groups, most psychosocial factors in bully group were not significantly different with those in nonbully group. Bully group was more influenced by internal psychic state, psychopathology and environmental factor than nonbully group.

Immunophenotypes of umbilical cord blood cells and GM-CSF mobilized peripheral blood cells / 대한임상병리학회지

BACKGROUDS: Different clinical outcomes of umbilical cord blood (CB) transplantation and GM-CSF mobilized peripheral blood stem cell (LB) transplantation stimulated our interests in immune characteristics of various stem cell sources so that we investigated immunophenotypes of mononuclear cells of twenty-six CB and ten LB cases and twenty normal adult peripheral blood (AB). METHODS: Two-color direct immunofluorescence analysis was performed using FACScan (Becton-Dickinson, USA) and monoclonal antibodies were CD34FITC/CD33PE, CD34FITC/CD45ROPE, CD34PE/CD45RAFITC, CD34PE/CD38FITC, CD4PE/CD45RAFITC, CD4FITC/CD45ROPE, CD8PE/CD45RAFITC, CD8PE/CD11bFITC, CD8PE/CD57FITC, CD8FITC/CD38PE (Serotec), and Simulset IMK lymphocyte kit (Becton-Dickinson). RESULTS: CD34+ cell frequencies of CB and LB were four times higher than that of AB. CD34+CD38- cell frequency was higher in CB, 41.5+/-28.1% compared to 20.1+/-15.8% in LB (P=0.0391) and frequencies of CD34+CD33+, CD34+CD45RA-, and CD34+CD45RO+ showed no difference. LB showed higher frequency of CD3+ and CD8+ cells and lower frequency of CD4+ and CD19+ cells compared to that of CB and AB. CD4+CD45RA+ na ve cell frequency was higher in CB (29.3%) compared to 2.3% of LB. CD4+CD45RO+ memory T lymphocyte frequency (8.5%) and CD8+CD57+ cell frequency (1.0%) were lowest in CB. CD8+CD45RA+ and CD8+CD11+ cell frequencies showed no difference. No statistically significant differences in CD34+ cell and its subsets and lymphocytes and their subsets were observed between normal vaginal delivered umbilical cord blood (VD) and Cesarean sectioned umbilical cord blood (CS), except CD8+CD11+ cell frequency was higher in VD (11+/-7.6%) compared to that (4.0+/-2.3%) in CS (P=0.0081). CONCLUSIONS: CB showed higher frequency of immature hematopoietic stem cell precursors and na ve lymphocytes compared to that of LB and AB and LB showed higher frequency of T lymphocytes and lower frequency of B lymphocytes compared to those of CB and AB, which might be the possible causes of the different clinical outcome. Study on the standard immunophenotypic values in various hematopoietic stem cell products using sufficient number of subjects is necessary for the guideline of clinical utilization.

A case of Wegener's granulomatosis complicated by diffuse pulmonary hemorrhage and thrombotic thrombocytopenic purpura

Wegener's granulomatosis is a distinct form of necrotizing granulomatous vasculitis which usually affects the kidneys and the upper and lower respiratory tracts. Unusual manifestations have also been reported, and these include colitis, urethritis and diabetes insipidus. We describe a case of Wegener's granulomatosis which presented with rapidly progressive renal insufficiency, sudden deafness, red eye, facial palsy, and complicated by uncommon manifestations that were diffuse pulmonary hemorrhage and thrombotic thrombocytopenic purpura.

A Case Report of Mantle Cell Lymphoma in Leukemic Phase / 대한임상병리학회지

Recently, we experienced a patient with a B cell leukemia which could not be classified as chronic lymphocytic leukemia (CLL), prolymphocytic leukemia (PL) or follicular lymphoma in leukemic phase. He is a 58-year-old male and suffered from progressive lymphadenopathy at both inguinal and cervical areas. His blood film showed lymphocytosis (79%), and the cells were heterogeneous in size and shape. Twenty-five percent of the lymphocytes had nuclear irregularities or cleavages unlike the lymphocytes of CLL In which the cells often shows monomorphic features, small size with scanty cytoplasm and round nucleus. The bone marrow trephine biopsy specimen showed a diffuse pattern of infiltration of atypical Iymphoid cells. The lymph node histology showed atypical lymphoid colls proliferated as wide mantles around non-neoplastic appearing germinal centers. The immunophenotype of circulating lymphocytes in peripheral blood showed strong reactivity with CD5, CDl9 and CD20 without expression of CD2, CD3, CD7, CD10, CD22 and CD23. Although this case resembled CLL, the laboratory features showed major differences, notably in the Peripheral blood morphology, histology Patterns and the membrane Phenotype. By combining these data, we diagnosed this case as a leukemic phase of mantle cell lymphoma.

Comparison of Broth Macrodilution and Microdilution in Testing of Minimal Inhibitory Concentration of Candida Species / 대한임상병리학회지

BACKGROUND: Candida species are one ol the major pathogens causing opportunistic infection. The need for a standardized. and clinically relevant method for antifungal susceptibility testing has become more increased because resistance to antifungal agents is now being reported with increasing frequency. The purposes of this study were to measure the minimal inhibitory concentrations actions amphotericin B, flucytosine and fluconazole of clinical isolates of Candida species, and to evaluate broth microdilution method as a more efficient test method, compared with NCCLS standard broth macrodilution. METHODS: The minlmal Inhibitory concentrations of C. parapsilosis ATCC 90018, C, albicans ATCC 90028 and 32 Candida isolates from Korea University Hospital were evaluated against amphotericin B, flucytosine and fluconazole by broth macrodilution and microdilution methods. RESULTS: The minimal inhibitory concentrations of Candida species ranged 0.25 to 1.0 microgram/mL for amphotericin B, 0.125 to 2.0 microgram/mL for flucytosine and 0.125 to 64 microgram/mL for fluconazole. The percent of agreement between NCCLS standard broth macrodilution and microdilution was highest when inoculum size was 0.5-2.5x103 CFU/mL and incubation time was 48 hours, and one fold dilution agreement was 100% for amphotericin B, 100% for flucytosine and 88-96% for fluconazole. CONCLUSIONS: This study showed good agreement of results of broth microdilution test with those of macrodilution, and suggested that broth microdilution method is an accessible and useful method and can be substituted for broth macrodilutlon method with the expectation of good results.