OBJECTIVE: To assess magnetic resonance imaging (MRI) features of coronary microembolization in a swine model induced by small-sized microemboli, which may cause microinfarcts invisible to the naked eye. MATERIALS AND METHODS: Eleven pigs underwent intracoronary injection of small-sized microspheres (42 microm) and catheter coronary angiography was obtained before and after microembolization. Cardiac MRI and measurement of cardiac troponin T (cTnT) were performed at baseline, 6 hours, and 1 week after microembolization. Postmortem evaluation was performed after completion of the imaging studies. RESULTS: Coronary angiography pre- and post-microembolization revealed normal epicardial coronary arteries. Systolic wall thickening of the microembolized regions decreased significantly from 42.6 +/- 2.0% at baseline to 20.3 +/- 2.3% at 6 hours and 31.5 +/- 2.1% at 1 week after coronary microembolization (p < 0.001 for both). First-pass perfusion defect was visualized at 6 hours but the extent was largely decreased at 1 week. Delayed contrast enhancement MRI (DE-MRI) demonstrated hyperenhancement within the target area at 6 hours but not at 1 week. The microinfarcts on gross specimen stained with nitrobluetetrazolium chloride were invisible to the naked eye and only detectable microscopically. Increased cTnT was observed at 6 hours and 1 week after microembolization. CONCLUSION: Coronary microembolization induced by a certain load of small-sized microemboli may result in microinfarcts invisible to the naked eye with normal epicardial coronary arteries. MRI features of myocardial impairment secondary to such microembolization include the decline in left ventricular function and myocardial perfusion at cine and first-pass perfusion imaging, and transient hyperenhancement at DE-MRI.
Animals ; Coronary Angiography/*methods ; Coronary Vessels/*pathology ; Disease Models, Animal ; Embolism/*pathology ; Female ; Heart/radiography ; Image Processing, Computer-Assisted ; Magnetic Resonance Imaging/*methods ; Microspheres ; Myocardial Contraction/physiology ; Myocardial Infarction/*pathology ; Myocardium/pathology ; Nitroblue Tetrazolium ; Staining and Labeling ; Swine ; Troponin T/blood ; Ventricular Function, Left

PURPOSE: Leukemic promyelocytes have the unique ability to undergo differentiation after exposure to retinoic acid and both differentiation and apoptosis after exposure to arsenic trioxide (ATO). Recent studies have shown that inhibition of Src family kinases (SFKs) resulted in enhancement of retinoic acid-induced myeloid differentiation. MATERIALS AND METHODS: In this study, we investigated the question of whether the SFK inhibitor PP2 enhanced the differentiation of NB4 cells when combined with ATO as well as when combined with all-trans-retinoic acid (ATRA). In addition, we attempted to determine the difference in retinoic acid-induced gene expression between cells treated with PP2 in combination with ATRA and in combination with ATO. RESULTS: SFK inhibitor PP2 induced significant enhancement of ATRA- or ATO-induced differentiation of NB4 cells. A significantly stronger synergistic effect was observed when PP2 was combined with ATRA than when combined with ATO. Flow cytometric analysis demonstrated a significant increase in CD11b-positive granulocytes up to 60.73% and 31.58%, respectively. These results were confirmed by nitroblue tetrazolium staining. These effects were not related to apoptosis. Results of Annexin-V-fluorescein staining revealed that PP2 combined with ATRA or PP2 combined with ATO did not induce apoptosis in NB4 cells. Retinoic acid-induced gene expression was different in both groups. Intercellular adhesion molecule-1 expression showed a significant increase in cells treated with PP2 in combination with ATRA, whereas cathepsin D expression showed a significant increase in cells treated with PP2 in combination with ATO. CONCLUSION: Our data showed that SFK inhibitor PP2 enhanced acute promyelocytic leukemia cell differentiation when combined with either ATRA or ATO with difference in activation of retinoic acid-induced genes.
Apoptosis ; Arsenic ; Arsenicals ; Cathepsin D ; Cell Differentiation ; Cell Line ; Gene Expression ; Granulocyte Precursor Cells ; Granulocytes ; Humans ; Intercellular Adhesion Molecule-1 ; Leukemia, Promyelocytic, Acute ; Nitroblue Tetrazolium ; Oxides ; Phosphotransferases ; Pyrimidines ; src-Family Kinases ; Tretinoin

BACKGROUND: The immunomodulatory effects of Korean mistletoe (Viscum album Coloratum) on the innate immune responses of eel (Anguilla japonica) were studied. METHODS: Mistletoe, Freund's complete adjuvant (FCA), or phosphate-buffered saline (PBS) as a control was injected into eel peritoneal cavities. RESULTS: Nitroblue tetrazolium (NBT)-positive cells in the head kidney of eel were significantly augmented by the second day post-injection of mistletoe. Reactive oxygen intermediates (ROI) were more produced in mistletoe-injected fish kidney leucocytes than in FCA-injected ones. The level of lysozyme activity in the serum of fish 2 days after injection with mistletoe was also significantly higher than that in the serum of the control fish. The optimal concentration of mistletoe in inducing the highest serum lysozyme activity was revealed to 500microgram/200 g of fish. In phagocytic activity assay, mistletoe-sensitized eel kidney phagocytes captured more zymosan than did the control fish. CONCLUSION: Korean mistletoe appeared to be a good activator of the non-specific immune responses of eel.
Eels ; Head Kidney ; Immunity, Innate ; Kidney ; Mistletoe ; Muramidase ; Nitroblue Tetrazolium ; Oxygen ; Phagocytes ; Zymosan

Chronic granulomatous disease (CGD) is a rare primary immunodeficiency in children caused by an abnormal function of the nicotinamide adenine dinucleotide phosphate (NADPH) oxidase in the phagocytic cells, which results in an increased susceptibility to severe bacterial and fungal infections. Prophylactic trimethoprim-sulfamethoxazole improves medium-term survival, but cannot prevent inflammatory sequelae. It still shows high morbidity and mortality. Bone marrow transplantation (BMT) is currently the only curative treatment for CGD. We report on a 29-month-old boy with CGD who was successfully treated with allogeneic BMT from an HLA-identical unrelated donors following a conditioning regimen consisting of busulfan and cyclophosphamide. One year after post-transplantation, the boy is in excellent clinical and hematological condition with complete chimerism and normal nitroblue tetrazolium (NBT) test.
Bone Marrow Transplantation ; Busulfan ; Child ; Child, Preschool ; Chimerism ; Cyclophosphamide ; Granulomatous Disease, Chronic* ; Humans ; Male ; Mortality ; NADP ; Nitroblue Tetrazolium ; Oxidoreductases ; Phagocytes ; Stem Cell Transplantation* ; Stem Cells* ; Trimethoprim, Sulfamethoxazole Drug Combination ; Unrelated Donors

BACKGROUND AND OBJECTIVES: Postnasal drip is one of the most common symptoms of chronic rhinosinusitis (CRS), and is the main cause of chronic cough. To evaluate the effect of upper airway inflammation defined as CRS on lower bronchial airway, we compared the cytology of nasal secretion (NS) and bronchoalveolar lavage fluid (BALF) of normal controls and patients with chronic rhinosinusitis accompanying with and without chronic cough normal controls. MATERIALS AND METHOD: Ten patients of CRS with postnasal drip were selected. Five of them had chronic cough and the others not. Five normal controls were selected. NS collected using Juhn's tymanic tap and BALF collected through fiberoptic bronchoscopy were diluted with dithiothreitol and PBS. These samples were centrifused and then cytospin slide was prepared. The cytology of the slides were evaluated under light microscope after Wright stain. To examine neutrophil activity, nitroblue tetrazolium dye (NBT) test was performd. Statistical analysis was performed using Mann-Whitney Rank Sum W test. RESULTS: In NS, there were no significant differences in the cell populations among coughing, noncoughing, and the normal control group. NBT positivity of coughing (34.2%) and noncoughing (31.5%) groups showed significantly higher than those of controls (8.6%). In BALF of coughing group, the population of macrophages (78.0%) was significantly lower than noncoughing (86.6%) and control (92.8%) groups, and population of lymphocytes (20.8%) was significantly higher than noncoughing (12.6%) and the control (6.4%) groups. In BALF of noncoughing group, the population macrophages was lower and those of lymphocytes were higher than the control group. CONCLUSION: These results suggest that CRS enhances increased local immune responses and decreased phagocytic activity of the lower airway. And chronic cough in patients with CRS is thought to be dependent on individual tolerance to cough provocation, not on aspiration of postnasal drip of discharge.
Bronchoalveolar Lavage Fluid* ; Bronchoalveolar Lavage* ; Bronchoscopy ; Cough* ; Dithiothreitol ; Humans ; Inflammation ; Lymphocytes ; Macrophages ; Neutrophils ; Nitroblue Tetrazolium ; Sinusitis

PURPOSE: It has been reported that the Nitroblue Tetrazolium(NBT) test is more accurate than the urine pH, leukocyte esterase and nitrite test as a screening test of urinary tract infection (UTI). The purpose of this study is to compare the NBT test with other screening tests and evaluate the clinical usefulness of the NBT test as a screening test. METHODS: We selected 298 results out of the 304 urine cultures which were performed from March, 1999 to July, 1999 and compared them with screening tests such as NBT, urine pH, leukocyte esterase and nitrite tests. We interpreted those results as the urinary tract infection when the screening results were urine NBT(+), pH(>6.5), leukocyte esterase(>or=++) and nitrite(+). RESULTS: Urine NBT, pH and leukocyte esterase tests showed the statistical significance in comparison with the urine culture results(chi-square tests; P<0.001, <0.05, <0.001), while urine nitrite tests did not show statistical significance. As time passing, the sensitivity of 10 min, 30 min, 60 min NBT test was increased to 35%, 72%, 80% respectively but the sensitivity of urine pH and leukocyte esterase was as low as 33%, 16% respectively. But, the specificity of NBT test was reduced from 93% to 53% as the time went by, while the specificity of urine pH and leukocyte esterase tests were as high as each 79%, 96% respectively. Urine NBT tests at 10 min and 30 min showed a higher positive and negative predictive value than those of the other screening tests. CONCLUSION: Urine NBT test as a screening test for UTI was more accurate than the urine pH, nitrite and leukocyte esterase tests. But we think that we should develop a more quick and precise screening test in the future, because of the long time it requires to perform it.
Child* ; Humans ; Hydrogen-Ion Concentration ; Leukocytes ; Mass Screening* ; Nitroblue Tetrazolium ; Sensitivity and Specificity ; Urinary Tract Infections* ; Urinary Tract*

BACKGROUND: Shiga toxin-producing Escherichia coli (STEC) was found in several serotypes of E coli including 0157 serotype. Sorbitol-MacConkey agar: may be useful for the detection of E. coli 0157, but is not helpful for the detection of sorbitol-fermenting STEC other than 0157. Moreover, some strains of E. colt 0157 can ferment sorbitol. In this study, in situ hybridization using DNA probe of shiga toxin was used for the isolation of STEC from the PCR-positive stool and -Sequenbe analysis of a part of shiga toxin gene was performed. METHODS: The stool was incubated in LB broth overnight and DNA was extracted from the culture fluid. Multiplex PCR was performed with primers for stxl and stx2 genes. Specimen showed PCR-positive was incubated on MacConkey agar and colonies were blotted with nitrocellulose membrane. Digoxigenin-labelled DNA probe for shiga toxin was made by PCR and the positive colonies were detected with anti-digoxigenin-alkaline phosphatase conjugate and nitroblue tetrazolium. Agglutination test with antisera was performed for the serotying and VTEC-RPLA kit was used for the toxin production. Sequence analysis of PCR products was performed with automatic sequence analyser. RESULTS: An stxl-negative, but stx2-positive PCR was observed in a three-year-old girl, who visited Kumi Hospital on July 19, 1999 complaining of vomiting and diarrhea. The positive colonies were isolated by in situ hybridization using stx2-specific DNA probe. The titers of stxl and stx2 by VTEC-RPLA test were negative and 1:64, respectively. Agglutination for the serotyping was not observed with all of the 0 antisera. 160-nucleotide sequence of stx2 of this isolate was identical with bacteriophage 933W (GenBank X07865), except for the change (T-C) of 957th nucleotide and amino acid sequence was identical each other. CONCLUSIONS: For the sensitive detection of STEC from the stool of patients with diarrhea, multiplex PCR is recommended with stxl- and stx2-specific primers. And in situ hybridization should be performed in PCR-positive specimen for the isolation of STEC. This method may be helpful for the detection of STEC as the causative microorganisms in food-borne outbreak.
Agar ; Agglutination ; Agglutination Tests ; Amino Acid Sequence ; Bacteriophages ; Collodion ; Diarrhea ; DNA ; Escherichia coli ; Female ; Gyeongsangbuk-do ; Humans ; Immune Sera ; In Situ Hybridization ; Membranes ; Multiplex Polymerase Chain Reaction ; Nitroblue Tetrazolium ; Polymerase Chain Reaction ; Sequence Analysis* ; Serotyping ; Shiga Toxin ; Shiga-Toxigenic Escherichia coli* ; Sorbitol ; Vomiting

BACKGROUND: Shiga toxin-producing Escherichia coli (STEC) was found in several serotypes of E coli including 0157 serotype. Sorbitol-MacConkey agar: may be useful for the detection of E. coli 0157, but is not helpful for the detection of sorbitol-fermenting STEC other than 0157. Moreover, some strains of E. colt 0157 can ferment sorbitol. In this study, in situ hybridization using DNA probe of shiga toxin was used for the isolation of STEC from the PCR-positive stool and -Sequenbe analysis of a part of shiga toxin gene was performed. METHODS: The stool was incubated in LB broth overnight and DNA was extracted from the culture fluid. Multiplex PCR was performed with primers for stxl and stx2 genes. Specimen showed PCR-positive was incubated on MacConkey agar and colonies were blotted with nitrocellulose membrane. Digoxigenin-labelled DNA probe for shiga toxin was made by PCR and the positive colonies were detected with anti-digoxigenin-alkaline phosphatase conjugate and nitroblue tetrazolium. Agglutination test with antisera was performed for the serotying and VTEC-RPLA kit was used for the toxin production. Sequence analysis of PCR products was performed with automatic sequence analyser. RESULTS: An stxl-negative, but stx2-positive PCR was observed in a three-year-old girl, who visited Kumi Hospital on July 19, 1999 complaining of vomiting and diarrhea. The positive colonies were isolated by in situ hybridization using stx2-specific DNA probe. The titers of stxl and stx2 by VTEC-RPLA test were negative and 1:64, respectively. Agglutination for the serotyping was not observed with all of the 0 antisera. 160-nucleotide sequence of stx2 of this isolate was identical with bacteriophage 933W (GenBank X07865), except for the change (T-C) of 957th nucleotide and amino acid sequence was identical each other. CONCLUSIONS: For the sensitive detection of STEC from the stool of patients with diarrhea, multiplex PCR is recommended with stxl- and stx2-specific primers. And in situ hybridization should be performed in PCR-positive specimen for the isolation of STEC. This method may be helpful for the detection of STEC as the causative microorganisms in food-borne outbreak.
Agar ; Agglutination ; Agglutination Tests ; Amino Acid Sequence ; Bacteriophages ; Collodion ; Diarrhea ; DNA ; Escherichia coli ; Female ; Gyeongsangbuk-do ; Humans ; Immune Sera ; In Situ Hybridization ; Membranes ; Multiplex Polymerase Chain Reaction ; Nitroblue Tetrazolium ; Polymerase Chain Reaction ; Sequence Analysis* ; Serotyping ; Shiga Toxin ; Shiga-Toxigenic Escherichia coli* ; Sorbitol ; Vomiting

This study was undertaken to investigate an involvement of nitroxergic innervation in gastric smooth muscle of rat. Isometric tension study, the measurement of single cell length, NADPH diaphorase stain of smooth muscle layers and neuronal nitric oxide synthase (nNOS) western blotting were performed. Sodium nitroprusside (SNP), a nitric oxide donor, relaxed the muscle strips precontracted by acetylcholine (ACh) in a concentration-dependent manner. Pretreatment of L-arginine decreased the contraction induced by electric field stimulation (EFS). Pretreatment of NG-nitro-L-arginine methyl ester (L-NAME), a NOS inhibitor, increased the EFS-induced contractions. LY 83583, a guanylate cyclase (GC) inhibitor, reversed the inhibitory actions of L-arginine on the muscle contractions. The effects of L-Arginine, L-NAME and LY 83583 on ACh-induced contractions were not significant. L-arginine reduced the EFS-induced contraction in circular muscle, whereas L-NAME enhanced the EFS-induced contraction in longitudinal strips. By EFS, the phasic contractions appeared approximately 20~25 seconds later. L-NAME significantly shortened the delay time to about 2~3 seconds. In single cell study, ACh contracted gastric smooth muscle cells, SNP relaxed the cells, and the latter also inhibited the ACh-induced contraction. LY 83583 enhanced the ACh-induced contraction and antagonized SNP-induced relaxation. NADPH diaphorase activity was assessed by a histochemistry, nitroblue tetrazolium (NTB) staining. Positive staining was observed in both circular and longitudinal muscle layers. L-arginine increased the staining, while L-NAME decreased the staining. Western blotting for nNOS proved the presence of nNOS in rat gastric smooth muscle. EFS and additional Ca2+ increased nNOS protein expression. These results suggest that in rat stomach, both circular and longitudinal muscle layers are innervated with nitroxergic nerves which relax the gastric smooth muscle via NO-cGMP pathway.
Acetylcholine ; Animals ; Arginine ; Blotting, Western ; Guanylate Cyclase ; Humans ; Muscle Contraction ; Muscle, Smooth* ; Myocytes, Smooth Muscle ; NADPH Dehydrogenase ; NG-Nitroarginine Methyl Ester ; Nitric Oxide ; Nitric Oxide Synthase Type I ; Nitroblue Tetrazolium ; Nitroprusside ; Rats* ; Relaxation ; Stomach ; Tissue Donors

Transition metal ions including Se2+, Cd2+, Hg2+ or Mn2+ have been thought to disturb the bone metabolism directly. However, the mechanism for the bone lesion is unknown. In this study, we demonstrated that MC3T3E1 osteoblasts, exposed to various transition metal ions; selenium, cadmium, mercury or manganese, generated massive amounts of reactive oxygen species (ROS). The released ROS were completely quenched by free radical scavengers-N-acetyl cysteine (NAC), reduced glutathione (GSH), or superoxide dismutase (SOD). First, we have observed that selenium (10 micrometer), cadmium (100 micrometer), mercury (100 micrometer) or manganese (1 mM) treatment induced apoptotic phenomena like DNA fragmentation, chromatin condensation and caspase-3-like cysteine protease activation in MC3T3E1 osteoblasts. Concomitant treatment of antioxidant; N-acetyl-L-cysteine (NAC), reduced-form glutathione (GSH), or superoxide dismutase (SOD), prevented apoptosis induced by each of the transition metal ions. Catalase or dimethylsulfoxide (DMSO) has less potent inhibitory effect on the apoptosis, compared with NAC, GSH or SOD. In line with the results, nitroblue tetrazolium (NBT) stain shows that each of the transition metals is a potent source of free radicals in MC3T3E1 osteoblast. Our data show that oxidative damage is associated with the induction of apoptosis in MC3T3E1 osteoblasts following Se2+, Cd2+, Hg2+ or Mn2+ treatment.
Acetylcysteine ; Apoptosis* ; Cadmium ; Catalase ; Chromatin ; Cysteine ; Cysteine Proteases ; Dimethyl Sulfoxide ; DNA Fragmentation ; Free Radicals ; Glutathione ; Ions ; Manganese ; Metabolism ; Metals ; Nitroblue Tetrazolium ; Osteoblasts* ; Reactive Oxygen Species ; Selenium ; Superoxide Dismutase