The present study was undertaken to investigate the influence of hypothermia on endothelium-independent vascular smooth muscle contractility and to determine the mechanism underlying the relaxation. Denuded aortic rings from male rats were used and isometric contractions were recorded and combined with molecular experiments. Hypothermia significantly inhibited fluoride-, thromboxane A2-, phenylephrine-, and phorbol ester-induced vascular contractions regardless of endothelial nitric oxide synthesis, suggesting that another pathway had a direct effect on vascular smooth muscle. Hypothermia significantly inhibited the fluoride-induced increase in pMYPT1 level and phorbol ester-induced increase in pERK1/2 level, suggesting inhibition of Rho-kinase and MEK activity and subsequent phosphorylation of MYPT1 and ERK1/2. These results suggest that the relaxing effect of moderate hypothermia on agonist-induced vascular contraction regardless of endothelial function involves inhibition of Rho-kinase and MEK activities.
Animals ; Fluorides ; Humans ; Hypothermia* ; Isometric Contraction ; Male ; Muscle, Smooth, Vascular ; Nitric Oxide ; Phosphorylation ; Rats ; Relaxation ; rho-Associated Kinases*

No abstract available.
Ductus Arteriosus, Patent

PURPOSE: To improve survival and/or to avoid radiotherapy, high dose chemotherapy (HDCT) with autologous peripheral blood stem cell transplantation (PBSCT) was given to patients with recurrent or high risk medulloblastoma (MB)/primitive neuroectodermal tumor (PNET) as well as patients younger than 3 years of age. METHODS: Six patients (3 recurrent, 1 high risk, 2 younger than 3 years; 5 MBs and 1 PNET) received single or double HDCT and PBSCT with or without immunotherapy using interleukin-2. Chemotherapeutic regimen in the first HDCT included cyclophosphamide (1,500 mg/m2/ day for 4 days) and melphalan (60 mg/m2/day for 3 days). Chemotherapeutic regimen in the second HDCT included carboplatin (400 mg/m2/day for 3 days), thiotepa (250 mg/ m2/day for 3 days), and etoposide (200 mg/m2/day for 3 days). RESULTS: Nine HDCTs were applied in 6 patients. Three double HDCTs were rescued with peripheral blood stem cells collected during single round leukapheresis. Rapid hematologic recovery occurred in 4 patients. Engraft failure occurred in 1 patient and delayed granulocyte recovery and platelet engraft failure occurred in 1 patient. Three patients who had minimal disease before HDCT had event free survival for 7~18 months after HDCT. Tumor relapsed 8 and 12 months after single HDCT in 2 patients among 3 patients with recurrent MB/PNET. One patient with recurrent MB died due to engraft failure and sepsis. CONCLUSION: HDCT with autologous PBSCT is expected to improve survival of patients with poor prognosis MB/PNET including younger patients less than 3 years. Subsequent trials with larger number of patients and long-term follow-up are needed.
Blood Platelets ; Carboplatin ; Cyclophosphamide ; Disease-Free Survival ; Drug Therapy* ; Etoposide ; Follow-Up Studies ; Granulocytes ; Humans ; Immunotherapy ; Interleukin-2 ; Leukapheresis ; Medulloblastoma ; Melphalan ; Neural Plate* ; Neuroectodermal Tumors* ; Neuroectodermal Tumors, Primitive ; Peripheral Blood Stem Cell Transplantation* ; Prognosis ; Radiotherapy ; Sepsis ; Stem Cells ; Thiotepa

This study was aimed to observe that extremely low frequency magnetic field (ELF-MF) may be relevant to changes of major neurotransmitters in rat brain. After the exposure to ELF-MF (60 Hz, 2.0 mT) for 2 or 5 days, we measured the levels of biogenic amines and their metabolites, amino acid neurotransmitters and nitric oxide (NO) in the cortex, striatum, thalamus, cerebellum and hippocampus. The exposure of ELF-MF for 2 or 5 days produced significant differences in norepinephrine and vanillyl mandelic acid in the striatum, thalamus, cerebellum and hippocampus. Significant increases in the levels of serotonin and 5-hydroxyindoleacetic acid were also observed in the striatum, thalamus or hippocampus. ELF-MF significantly increased the concentration of dopamine in the thalamus. ELF-MF tended to increase the levels of amino acid neurotransmitters such as glutamine, glycine and gamma -aminobutyric acid in the striatum and thalamus, whereas it decreased the levels in the cortex, cerebellum and hippocampus. ELF-MF significantly increased NO concentration in the striatum, thalamus and hippocampus. The present study has demonstrated that exposure to ELF-MFs may evoke the changes in the levels of biogenic amines, amino acid and NO in the brain although the extent and property vary with the brain areas. However, the mechanisms remain further to be characterized.
Animals ; Biogenic Amines ; Brain ; Cerebellum ; Dopamine ; Glutamine ; Glycine ; Hippocampus ; Magnetic Fields* ; Neurotransmitter Agents* ; Nitric Oxide ; Norepinephrine ; Rats ; Serotonin ; Thalamus

Malaria is a global disease affecting a large portion of the world’s population. Although vaccines have recently become available, their efficacies are suboptimal. We generated virus-like particles (VLPs) that expressed either apical membrane antigen 1 (AMA1) or microneme-associated antigen (MIC) of Plasmodium berghei and compared their efficacy in BALB/c mice. We found that immune sera acquired from AMA1 VLP- or MIC VLP-immunized mice specifically interacted with the antigen of choice and the whole P. berghei lysate antigen, indicating that the antibodies were highly parasite-specific. Both VLP vaccines significantly enhanced germinal center B cell frequencies in the inguinal lymph nodes of mice compared with the control, but only the mice that received MIC VLPs showed significantly enhanced CD4+ T cell responses in the blood following P. berghei challenge infection. AMA1 and MIC VLPs significantly suppressed TNF-α and interleukin-10 production but had a negligible effect on interferon-γ. Both VLPs prevented excessive parasitemia buildup in immunized mice, although parasite burden reduction induced by MIC VLPs was slightly more effective than that induced by AMA1. Both VLPs were equally effective at preventing body weight loss. Our findings demonstrated that the MIC VLP was an effective inducer of protection against murine experimental malaria and should be the focus of further development.

PURPOSE: Long-term survivors of childhood cancer appear to have an increased risk for the metabolic syndrome, subsequent type 2 diabetes and cardiovascular disease in adulthood compared to healthy children. The purpose of this study was to investigate the frequency of the metabolic syndrome and associated factors in childhood cancer survivors at a single center in Korea. METHODS: We performed a retrospective review of medical records of 98 childhood cancer survivors who were diagnosed and completed anticancer treatment at Samsung Medical Center, Seoul, Korea between Jan. 1996 and Dec. 2007. Parameters of metabolic syndrome were evaluated between Jan. 2008 and Dec. 2009. Clinical and biochemical findings including body fat percentage were analyzed. RESULTS: A total of 19 (19.4%) patients had the metabolic syndrome. The median body fat percentage was 31.5%. The body mass index and waist circumference were positively correlated with the cranial irradiation dose (r=0.38, P<0.001 and r=0.44, P<0.00, respectively). Sixty-one (62.2%) patients had at least one abnormal lipid value. The triglyceride showed significant positive correlation with the body fat percentage (r=0.26, P=0.03). The high density lipoprotein cholesterol showed significant negative correlation with the percent body fat (r=-0.26, P=0.03). CONCLUSION: Childhood cancer survivors should have thorough metabolic evaluation including measurement of body fat percentage even if they are not obese. A better understanding of the determinants of the metabolic syndrome during adolescence might provide preventive interventions for improving health outcomes in adulthood.
Adipose Tissue ; Adolescent ; Body Composition ; Body Mass Index ; Cardiovascular Diseases ; Child ; Cholesterol ; Cholesterol, HDL ; Cranial Irradiation ; Humans ; Korea ; Lipoproteins ; Medical Records ; Retrospective Studies ; Survivors ; Waist Circumference

Transplantation of marrow-derived mesenchymal stem cells (MSCs), expanded by culture in addition to whole bone marrow, has been shown to enhance engraftment of human hematopoietic stem cells (HSCs). Our hypothesis was that there might be an optimum ratio range that could enhance engraftment. We examined the percent donor chimerism according to the ratio of HSCs to MSCs in non-obese diabetic/severe combined immunodeficiency (NOD/SCID) mice. We tested a series of ratios of co-transplanted CD34+-selected bone marrow cells, and marrow-derived MSCs into sublethally irradiated NOD/SCID mice. In all experiments, 1 x 10(5) bone marrow derived human CD34+ cells were administered to each mouse and human MSCs from different donors were infused concomitantly. We repeated the procedure three times and evaluated engraftment with flow cytometry four weeks after each transplantation. Serial ratios of HSCs to MSCs were 1:0, 1:1, 1:2 and 1:4, in the first experiment, 1:0, 1:1, 1:2, 1:4 and 1:8 in the second and 1:0, 1:1, 1:4, 1:8 and 1:16 in the third. Cotransplantation of HSCs and MSCs enhanced engraftment as the dose of MSCs increased. Our results suggest that the optimal ratio of HSCs and MSCs for cotransplantation might be in the range of 1:8-1:16; whereas, an excessive dose of MSCs might decrease engraftment efficiency.
Middle Aged ; Mice, SCID ; Mice, Inbred NOD ; Mice ; Mesenchymal Stem Cells/*cytology ; Mesenchymal Stem Cell Transplantation/*methods ; Humans ; Hematopoietic Stem Cells/*cytology ; Hematopoietic Stem Cell Transplantation/*methods ; Graft Survival/*physiology ; Cells, Cultured ; Cell Count ; Animals ; Adult

Transplantation of marrow-derived mesenchymal stem cells (MSCs), expanded by culture in addition to whole bone marrow, has been shown to enhance engraftment of human hematopoietic stem cells (HSCs). Our hypothesis was that there might be an optimum ratio range that could enhance engraftment. We examined the percent donor chimerism according to the ratio of HSCs to MSCs in non-obese diabetic/severe combined immunodeficiency (NOD/SCID) mice. We tested a series of ratios of co-transplanted CD34+-selected bone marrow cells, and marrow-derived MSCs into sublethally irradiated NOD/SCID mice. In all experiments, 1 x 10(5) bone marrow derived human CD34+ cells were administered to each mouse and human MSCs from different donors were infused concomitantly. We repeated the procedure three times and evaluated engraftment with flow cytometry four weeks after each transplantation. Serial ratios of HSCs to MSCs were 1:0, 1:1, 1:2 and 1:4, in the first experiment, 1:0, 1:1, 1:2, 1:4 and 1:8 in the second and 1:0, 1:1, 1:4, 1:8 and 1:16 in the third. Cotransplantation of HSCs and MSCs enhanced engraftment as the dose of MSCs increased. Our results suggest that the optimal ratio of HSCs and MSCs for cotransplantation might be in the range of 1:8-1:16; whereas, an excessive dose of MSCs might decrease engraftment efficiency.
Middle Aged ; Mice, SCID ; Mice, Inbred NOD ; Mice ; Mesenchymal Stem Cells/*cytology ; Mesenchymal Stem Cell Transplantation/*methods ; Humans ; Hematopoietic Stem Cells/*cytology ; Hematopoietic Stem Cell Transplantation/*methods ; Graft Survival/*physiology ; Cells, Cultured ; Cell Count ; Animals ; Adult

BACKGROUND AND OBJECTIVES: Vascular endothelial growth factor (VEGF) is a potent endothelial cell-specific mitogen. This study was undertaken to test the hypothesis that the neointima hyperplasia induced by a balloon injury is inhibited by blocking VEGF and VEGF receptor-1 (VEGFR-1) with anti-VEGF peptides. Materials and Methods: Anti-VEGF RRKRRR peptide (dRK6) and anti-VEGFR-1 peptide (anti-flt-1) were synthesized at Pohang University of Science and Technology, Korea. Male Sprague-Dawley rats, weighing 300-350 g, were subcutaneously injected 0.5 mg/kg of dRK6 or 0.5 mg/kg of anti-flt-1, dissolved in phosphate buffer solution, 2 days before induction of a carotid balloon-injury, and then daily in the same manner post carotid balloon injury for 2 weeks. RESULTS: Neointima formation was suppressed in both the dRK6 and anti-flt-1 groups compared to that in the untreated controls at 2 weeks post carotid balloon-injury (neointimal area; control group 0.44+/-0.09 mm2, dRK6 group 0.25+/-0.05 mm2, anti-flt-1 group 0.19+/-0.05 mm2, p<0.01). Anti-flt-1 peptide and dRK6 reduced the numbers of proliferative bromodeoxyuridine-labeled cells in the neointima (control group 16.4+/-10.6%, dRK6 group 3.7+/-2.1%, anti-flt-1 group 5.9+/-3.4%, p<0.05). In addition, an inflammatory response, as determined by monocyte chemoattractant protein-1 and interleukin-6 upregulation, which was evident in the controls, was inhibited by both dRK6 and anti-flt-1. CONCLUSION: This study suggests anti-vascular endothelial growth factor peptides can reduce the inflammation and neointima formation in balloon injured rat carotid arteries.
Animals ; Carotid Arteries* ; Carotid Artery Injuries ; Chemokine CCL2 ; Endothelial Growth Factors* ; Gyeongsangbuk-do ; Humans ; Hyperplasia ; Inflammation ; Interleukin-6 ; Korea ; Male ; Neointima* ; Peptides* ; Rats* ; Rats, Sprague-Dawley ; Up-Regulation ; Vascular Endothelial Growth Factor A ; Vascular Endothelial Growth Factor Receptor-1 ; Vascular Endothelial Growth Factors

PURPOSE: Current pediatric cancer research requires an organized pediatric tumor tissue bank with standardized guidelines for preparation and storage of human tumor tissue samples, white cells, serum, genomic DNA, RNA, cDNA and proteins.. Our institution established and managed pediatric tumor tissue bank for the last one year, and we want to present an overview of our experiences and guidelines. METHODS: From leukemia patients, peripheral blood and bone marrow aspirates were collected at initial diagnosis. Leukemic cells were prepared by Ficoll density-gradient centrifugation and stored at 196oC liquid nitrogen. For solid tumors, tissue cultures were performed as soon as possible after surgical excision or needle biopsy. Serum free media and primary cultured cells were collected and stored at 20degrees C and at 196degrees C, respectively. Genomic DNA, RNA and cDNA were isolated from leukemic cells and cultured solid tumor cells, and stored at 20degrees C. We also isolated genomic DNA from white blood cells of solid tumor patients and stored at 20degrees C. Finally we collected serum samples from all pediatric cancer patients at diagnosis and stored at 20degrees C. RESULTS: Among the 41 cases of leukemia and 100 cases of solid tumor patients who were diagnosed at department of pediatrics, Samsung Medical Center, from August 2000 to July 2001, 26 cases (63%) of leukemia and 59 cases (59%) of solid tumor patients were registered to Pediatric Tumor Tissue Bank. Primary cell cultures were performed in 21 cases of solid tumors and were successful in 19 cases (90%). The isolated genomic DNA, RNA and cDNA were all in high quality confirmed by electrophoresis in agarose gel. CONCLUSION: The problem of tissue sample size obtained by needle biopsy could be overcome by primary cell cultures. For the effective management of pediatric tumor tissue bank, fresh tissue collection with active cooperation of surgeons, organized personnel structure, and multidisciplinary standardized guidelines are necessary.
Biopsy, Needle ; Bone Marrow ; Cell Culture Techniques ; Cells, Cultured ; Centrifugation ; Culture Media, Serum-Free ; Diagnosis ; DNA ; DNA, Complementary ; Electrophoresis ; Ficoll ; Humans ; Leukemia ; Leukocytes ; Nitrogen ; Pediatrics ; Primary Cell Culture ; RNA ; Sample Size ; Sepharose ; Tissue Banks*