BACKGROUND: Chemotaxis is one of the cardinal functions of leukocytes, which enables them to be recruited efficiently to the right place at the right time. Analyzing chemotactic activities is important not only for the study on leukocyte migration but also for many other applications including development of new drugs interfering with the chemotactic process. However, there are many technical limitations in the conventional in vitro chemotaxis assays. Here we applied a new optical assay to investigate chemotactic activities induced in differentiated HL-60 cells. METHODS: HL-60 cells were stimulated with 0.8% dimethylformamide (DMF) for 4 days. The cells were analyzed for morphology, flow cytometry as well as chemotactic activities by a time-lapse videomicroscopic assay using a chemotactic microchamber bearing a fibronectin-coated cover slip and an etched silicon chip. RESULTS: Videomicroscopic observation of the real cellular motions in a stable concentration gradient of chemokines demonstrated that HL-60 cells showed chemotaxis to inflammatory chemokines (CCL3, CCL5 and CXCL8) and also a homeostatic chemokine (CXCL12) after DFM-induced differentiation to granulocytic cells. The cells moved randomly at a speed of 6.99+/-1.24 micrometer/min (n=100) in the absence of chemokine. Chemokine stimulation induced directional migration of differentiated HL-60 cells, while they still wandered very much and significantly increased the moving speeds. CONCLUSION: The locomotive patterns of DMF-stimulated HL-60 cells can be analyzed in detail throughout the course of chemotaxis by the use of a time-lapse videomicroscopic assay. DMF-stimulated HL-60 cells may provide a convenient in vitro model for chemotactic studies of neutrophils.
Chemokines ; Chemotaxis ; Dimethylformamide ; Flow Cytometry ; HL-60 Cells* ; Humans ; Leukocytes ; Microscopy, Video ; Neutrophils ; Silicon

Progress in medical science and cell biology has resulted in the transplantation of human cells and tissues from on human into another, facilitating reproduction and the restoration of form and function, as well as enhancing the quality of life. For more than 40 years, society has recognized the medical and humanitarian value of donation and transplanting organs and tissues. The standard operating procedures of hard tissues reflect the collective expertise and conscientious efforts of tissue bank professionals to provide a foundation for the guidance of tissue banking activities. Procurement of allograft tissues from surgical bone donors is a part of tissue banking. During the past decades the use of bone allografts has become widely accepted for the filling of skelectal defects in a variety of surgical procedures. In particular in the field of orthopaedic and oral and maxillofacial surgery the demand for allografts obtained from either living or post-mortem donors has increased. Hospital-based tissue banks mainly retrieve allografts from living donors undergoing primary total hip replacement for osteoarthritis or hemi arthroplasty for hip fractures and orthgnatic surgery such as angle reduction. Although bone banks have existed for many years, the elements of organized and maintaining a hospital bone bank have not been well documented. The experience with a tissue bank at Korea Tissue Bank(KTB) between 2001 and 2004 provides a model of procurement, storage, processing, sterilization and documentation associated with such a facility. The following report describes the standard operating procedures of hard tissues such as femoral head obtained from living donors.
Allografts* ; Arthroplasty, Replacement, Hip ; Bone Banks ; Head* ; Hemiarthroplasty ; Hip Fractures ; Humans ; Korea ; Living Donors* ; Osteoarthritis ; Quality of Life ; Reproduction ; Sterilization ; Surgery, Oral ; Tissue Banks ; Tissue Donors

PURPOSE: We aimed to study the feasibility of arsenic trioxide as a treatment of neuroblastoma which has the ability to differentiate into nonmalignant cells like acute promyelocytic leukemia. METHODS: To determine the effects of arsenic trioxide in various concentrations and exposure time on the survivial of neuroblastoma cell lines, SH-SY5Y and SK-N-AS cells were cultured in RPMI 1640 media with 1 to 20muM concentration of arsenic trioxide. Apoptosis was measured with flow cytometry by staining with 7-aminoactinomycin D. Cell cycle was assessed by monitoring the DNA contents by flow cytometry. Arsenic trioxide induced cell morphologic changes were also observed with May-Grunwald-Giemsa stain under a light microscope. RESULTS: Arsenic trioxide induced apoptosis in SH-SY5Y cells earlier in the same concentration and to a more severe degree with the same exposure time than in HL-60 cells. The apoptosis induced by arsenic trioxide was steeply increased to 79.3 10.1% at 24 hours and then maintained a plateau on 20muM concentration, while increasing steadily to 40.2 6.5% until 72 hours on 5muM concentration. The proliferating cell proportion in S/G2/M phase was decreased with arsenic trioxide concentration and with exposure time in both SH-SY5Y and HL-60 cells, especially more so with the SH-SY5Y cells. The cellularity was decreased and more apoptotic cells could be observed in the arsenic trioxide treatment group than in untreated control group. CONCLUSION: As in acute promyelocytic leukemic cells, arsenic trioxide induced apoptosis and cell cycle arrest of proliferating phase in neuroblastoma cells.
Apoptosis ; Arsenic* ; Cell Cycle ; Cell Cycle Checkpoints ; Cell Line ; DNA ; Flow Cytometry ; HL-60 Cells ; Humans ; Leukemia, Promyelocytic, Acute ; Neuroblastoma*

Immunohistochemical staining was used to evaluate the presence of nitricoxide synthase (NOS) family enzymes in the rat penis and major pelvic ganglion.We used three kinds of antisera which are brain(neuronal), endothelial cell andmacrophge(inducible) enzymes of NOS. Many brain NOS positive neurons wereobserved in the major pelvic ganglion, especially in the initial portion of thecavenous nerve. In the penile erectile tissue, many delicate, beaded or linearbrain NOS-positive fibers were observed on the bundle of smooth muscles,adventitia of helicine artery and wall of cavernous space. Brain NOS-positiveimmunoreactivity were also seen on the encapsulated nerve ending and adventitiaof dorsal vessels of penis. Endothelial cell NOS immunoreactivity was exclusivelyin the urethral epithelium of corpus spongiosum. There was no inducible NOSimmunoreactivity in rat penis and major pelvic ganglion. These results suggestthat NO related to penile erection in rat was made from brain NOS rather thanendothelia1 cell or inducible NOS.
Animals ; Arteries ; Brain ; Endothelial Cells ; Epithelium ; Ganglion Cysts* ; Humans ; Immune Sera ; Male ; Nerve Endings ; Neurons ; Nitric Oxide Synthase* ; Nitric Oxide* ; Penile Erection ; Penis* ; Protein Isoforms* ; Rats*

Allografts ; Bacteria ; Bacteriology ; Communicable Diseases ; Gamma Rays ; Humans ; Incidence ; Korea ; Radiation, Ionizing ; Sterilization ; Tendons ; Tissue Banks ; Tissue Donors

BACKGROUND: The cell-mediated immune reaction to tuberculosis infection involves a complex network of cytokines. The extent of inflammation, tissue damage and severity of the disease suggested to be determined by the balance between extent and duration of the proinflammatory cytokine response versus those of the suppressive cytokines. The systemic cytokine response in pathogenesis of tuberculosis can be assessed by measuring serum cytokine levels. METHOD: Serum interleukin-1 beta(IL-1 ), IL-2, IL-4, IL-6, IL-10, IL-12(p40), tumor necrosis factor-alpha(TNF-alpha), interferon-gamma(IFN-gamma) and transforming growth factor-beta(TGF-beta) levels were measured in 83 patients with pulmonary tuberculosis, 10 patients with endobronchial tuberculosis before treatment and 20 healthy subjects by using a sandwich ELISA. In patients with pulmonary tuberculosis, they were divided into mild, moderate and far advanced group according to the severity by ATS guidelines. To compare with those of pretreatment levels, we measured serum IL-1beta, IL-2, IL-4, IL-6, IL-10, IL-12(p40), TNF-alpha, IFN-gamma and TGF-beta levels in 45 of 83 patients with pulmonary tuberculosis after 2 and 6 months of treatment. RESULTS: 1) In sera of patients with active pulmonary tuberculosis(n=83), IL-1beta, IL-6(p<0.05), TNF-alpha, and IFN-gamma were elevated and TGF-beta was decreased comparing to control. IL-2, IL-12(p40), IL-4 and IL-10 were similar between the patients with tuberculosis and control. 2) In endobronchial tuberculosis, IL-6 and TNF-alpha were elevated and TGF-beta was decreased comparing to control. IL-12(p40) seemed to be elevated comparing to pulmonary tuberculosis. 3) Far advanced tuberculosis showed markedly elevated IL-6 and IFN-gamma level(p<0.05). 4) The significant correlations were noted between IL-1, IL-6 and TNF-alpha and between IL-12, IL-2 and IL-4(p<0.01). 5) After 2 and 6 months of standard treatment, the level of IL-6 and IFN-gamma was significantly decreased(p<0.05). CONCLUSION: These results showed that an altered balance between cytokines is likely to be involved in the extent of inflammation, tissue damage and severity of the disease tuberculosis. But, it should be considered diversities of cytokine response according to type of tuberculosis and immunity in clinical application and interpreting future studies.
Cytokines ; Enzyme-Linked Immunosorbent Assay ; Humans ; Inflammation ; Interleukin-1 ; Interleukin-10 ; Interleukin-12 ; Interleukin-2 ; Interleukin-4 ; Interleukin-6 ; Korea* ; Necrosis ; Transforming Growth Factor beta ; Tuberculosis* ; Tuberculosis, Pulmonary ; Tumor Necrosis Factor-alpha

BACKGROUND: A range of well characterized materials are needed for validating the performance of hepatitis B surface antigen (HBsAg) immunoassays. These materials are purchased currently from overseas manufacturers at a high cost and with limited quantity. This study was conducted to establish an HBsAg low titer performance panel for use as a national standard for validation of HBsAg immunoassays in Korea. METHODS: 476 plasma units reactive on blood donor screening were collected HBsAg was tested using 3 enzyme immunoassays (EIA) and 1 chemiluminescence immunoassay (CIA). Units reactive on the CIA assay or on 2 or more immunoassays were subjected to hepatitis B virus (HBV) DNA quantification, HBV genotyping and subtyping. Units reactive on HBV DNA quantification were confirmed for HBsAg by neutralization. Candidates for the panel were subjected to a collaborative study performed at 7 laboratories using 7 immunoassays. RESULTS: Eleven HBsAg positive units were selected for the low titer performance panel based on HBsAg immunoassay, HBV DNA quantification, HBV genotyping and subtyping results. The range of the HBsAg concentration of the panel members was 0.05~1.28 IU/mL. Two HBsAg negative units were also included as negative controls. CONCLUSION: As a result of this study, a low titer performance panel [KFDA standard (08/028); HBsAg low titer performance panel (BTRL HBV/LP)] for validation of HBsAg immunoassays has been established as a Korean national standard. Use of this panel will improve performance assessment of HBsAg immunoassays. Because the performance of immunoassays cannot be assessed properly with a limited number of panels, continuous efforts are needed to develop a range of performance panels.
Blood Donors ; DNA ; Hepatitis ; Hepatitis B ; Hepatitis B Surface Antigens ; Hepatitis B virus ; Humans ; Immunoassay ; Immunoenzyme Techniques ; Luminescence ; Mass Screening ; Plasma

Azathioprine (AZA) treatment in transplant or autoimmune patients and subsequent appearance squamous cell carcinomas at various sites, particularly skin and cervix, has shown a close relationship. However, it remains uncertain whether this is true for the patients with Crohn's disease. We report a case of squamous cell carcinoma of the breast occurred in a 35-year-old female with Crohn's disease taking AZA. She was first diagnosed with Crohn's disease 10 years ago and has taken AZA with 5-aminosalicylic acid (5-ASA) on regular follow up in gastrointestinal department for 9 years. She had no family history of breast cancer. She visited breast cancer clinic due to incidentally found right breast mass. A mastectomy on the right breast was performed and 6.3x5.5 cm mass was removed. The mass was microscopically proven to be poorly differentiated squamous cell carcinoma with focal keratin pearl formation. At age of 25, she was first diagnosed with active Crohn's disease. 5-ASA and corticosteroid induced remission. Then, steroid was tapered off and AZA was maintained at 1 mg/kg due to leukopenia at higher dose. She stopped taking AZA at her discretion during her two pregnancies and reported total of 67 months of AZA medication on her breast cancer diagnosis.
Adult ; Azathioprine/*therapeutic use ; Breast Neoplasms/*diagnosis/pathology/therapy ; Carcinoma, Squamous Cell/*diagnosis/pathology/therapy ; Colonoscopy ; Combined Modality Therapy ; Crohn Disease/*drug therapy ; Female ; Humans ; Immunosuppressive Agents/*therapeutic use ; Mesalamine/therapeutic use ; Positron-Emission Tomography

Thyroid carcinomas are uncommon in childhood and adolescence. The aim of this study was to analyze clinical features and clinical outcomes of thyroid cancer in the pediatric population treated in the Yonsei University Health System. From September 1982 to June 2009, 90 patients (75 females, 15 males; female:male ratio of 5:1) with differentiated thyroid carcinoma were identified in our institute. The mean age at diagnosis was 15.8 yr old (range 4.8-19.9 yr). Cervical masses were most common clinical manifestations at diagnosis in 65 patients (72.2%). Forty-two patients underwent less than total thyroidectomy and 18 patients underwent total thyroidectomy. Thirty patients (33.3%) had lateral neck lymph node metastasis and seven patients (7.8%) had lung metastasis at the time of surgery. Among the 90 patients, recurrence occurred in 14 patients (15.5%). Mean follow-up period for patients with differentiated thyroid carcinoma was 81.6 months (13-324 months). No patients died of differentiated thyroid carcinoma. Patients with differentiated thyroid carcinoma who were < 20-yr-of-age were present with aggressive local disease and a high frequency of lymph node and distant metastasis. It is recommended that pediatric thyroid cancer should be managed mostly using proper surgical approach with thyroidectomy and lymph node dissection when indicated.
Adolescent ; Age Factors ; Carcinoma/*pathology/surgery ; Child ; Child, Preschool ; Female ; Follow-Up Studies ; Hospitals, University ; Humans ; Iodine Radioisotopes/therapeutic use ; Lung Neoplasms/diagnosis/secondary ; Lymph Node Excision ; Lymph Nodes/surgery ; Lymphatic Metastasis ; Male ; Recurrence ; Survival Rate ; Thyroid Neoplasms/*pathology/radiotherapy/surgery ; Thyroidectomy ; Young Adult

PURPOSE: Neuroblastoma is a common tumor in childhood, and generally exhibits heterogeneity and a malignant progression. MYCN expression and amplification profiles frequently correlate with therapeutic prognosis. Although it has been reported that MYCN silencing causes differentiation and apoptosis in human neuroblastoma cells, MYCN expression influences the cytotoxic potential of chemotherapeutic drugs via the deregulation of the cell cycle. STI-571 may constitute a promising therapeutic agent against neuroblastoma, particularly in cases in which c-Kit is expressed preferentially in MYCN-amplified neuroblastoma. MATERIALS AND METHODS: To determine whether STI-571 exerts a synergistic effect on cytotoxicity with MYCN expression, we assessed apoptotic cell death and cell cycle distribution after 72 h of exposure to STI-571 with or with out treatment of SK-N-BE(2) neuroblastoma cells with MYCN siRNA. RESULTS: MYCN siRNA-treated SK-N-BE(2) cells did not affect apoptosis and cells were arrested in G0/G1 phase after STI-571 treatment. CONCLUSIONS: siRNA therapy targeted to MYCN may not be effective when administered in combination with STI-571 treatment in cases of neuroblastoma. Therefore, chemotherapeutic drugs that target S or G2-M phase may prove ineffective when applied to cells arrested in the G0/1 phase as the result of MYCN knockdown and STI-571 treatment.
Apoptosis ; Benzamides ; Cell Cycle ; Cell Death ; Cell Line ; Humans ; Neuroblastoma ; Piperazines ; Population Characteristics ; Prognosis ; Pyrimidines ; Imatinib Mesylate ; RNA, Small Interfering