PURPOSE: A urea breath test (UBT) using C-14 or C-13 has been developed for identifying Helicobacter (H) pylori infection on the basis of urease production with release of labeled CO2. We investigated if the C-14 and C-13 UBT have the difference to reflect the presence and degree of H. pylori infection detected by gastroduodenoscopic biopsies (GBx) in the same patients. Materials and METHODS: Thirty eight patients (M:F=28:10, age 53.4+/-13.0 yrs) with upper gastrointestinal symptoms such as indigestion, gastric fullness or pain consecutively underwent C-14 UBT, GBx and C-13 UBT within one week before medications. For the C-14 UBT, a single breath sample was collected at 10 minutes after ingestion of C-14 urea (37 KBq) capsule and counting was done in a liquid scintillation counter for 1 minute, and the results were classified as positive (> or =200 dpm), intermediate (50~199 dpm) or negative (<50 dpm). For the C-13 UBT, the results were classified as positive (> or =2.5 permill) or negative (<2.5 permill). The results of GBx with Giemsa stain were graded 0 (normal) to 4 (diffuse) according to the distribution of H. pylori by the Wyatt method. We compared C-14 UBT and C-13 UBT results with GBx grade as a gold standard. RESULTS: The prevalence of H. pylori infection by GBx with Giemsa stain was 25/38 (65.8%). In the assessment of the presence of H. pylori infection, the C-14 UBT global performance yielded sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV) and accuracy of 92.0%, 92.3%, 95.8%, 91.7% and 92.1%, respectively. However, the C-13 UBT had sensitivity, specificity, PPV, NPV and accuracy of 96.0%, 84.6%, 92.3%, 91.7% and 92.1%, respectively. The more significant correlation in C-14 than C-13 UBT (r=0.948 vs r=0.819, p<0.001) was found between the value of UBT and the grade of distribution of H. pylori infection. CONCLUSION: We conclude that the diagnostic performance between C-14 and C-13 UBT to detect H. pylori infection is not significantly different, but the value of C-14 UBT more significantly reflects the degree of bacterial distribution.
Azure Stains ; Biopsy ; Breath Tests ; Dyspepsia ; Eating ; Helicobacter ; Helicobacter pylori ; Humans ; Organothiophosphorus Compounds ; Prevalence ; Scintillation Counting ; Sensitivity and Specificity ; Urea ; Urease

PURPOSE: To investigate the expression of IRF-1, IRF-7 and iNOS in the mice model of Pseudomonas aeruginosa keratitis. The enzymatic activity of iNOS and its expression were also investigated. METHODS: With western blot analysis, the protein expression of IRF-1, IRF-7 (at 24 hours), and iNOS (at 12 hours and 24 hours) were evaluated in the mouse model of P. aeruginosa keratitis. iNOS enzymatic activity was determined with a scintillation counter. IRF-1 and IRF-7 expression were localized with immunofluorescent labeling. The wounded control group was given the same corneal wound without bacterial inoculation, and the fellow eyes served as normal controls. RESULTS: Expression of IRF-1, IRF-7 and iNOS was highly upregulated in corneas with P. aeruginosa keratitis compared to normal or wounded corneas. iNOS enzymatic activity also was higher in infected than normal corneas. In wounded corneas, NOS2 expression and activity slightly increased at 12 hours after the infection. Intense IRF-1 immunopositivity was seen in the epithelial layer of infected corneas. Some corneal stromal cells and endothelial cells showed moderate positive labeling in infected corneas. IRF-7 showed intense labeling in the epithelial layer and endothelial cells of normal as well as infected corneas. Increased IRF-7 labeling was observed in epithelial cells in the ulcerated region of infected corneas. CONCLUSIONS: These results suggest that IRF-1, IRF-7 and iNOS may play a regulatory role in the immune responses and wound healing process in P. aeruginosa keratitis.
Animals ; Blotting, Western ; Cornea* ; Endothelial Cells ; Epithelial Cells ; Keratitis* ; Mice* ; Pseudomonas aeruginosa* ; Pseudomonas* ; Scintillation Counting ; Stromal Cells ; Ulcer ; Wound Healing ; Wounds and Injuries

We established an in vitro experimental system using the following procedure. We first introduced tritium-labeled norepinephrine ([3H]-NE) into PC12 cells. The [3H]-NE incorporated-PC12 cells were stimulated by a high concentration (60 mM) of K+ buffer during 12 minutes. Then, we collected 100 microliter supernatant and counted the amount of [3H]-NE release from PC12 cells with a scintillation counter. After screening fungal, Streptomyces spp. or bacterial product using this experimental sytem, we obtained FS390 from Streptomyces spp. which inhibited [3H]-NE release from PC12 cells. FS390 also inhibits the release of ATP as a neurotransmitter of PC12 cells and rat cortical neurons. The inhibitory effect was seen even when the PC12 cells were treated with low K+ buffer containing ionomycin (1 micrometer) as an ionopore. This result suggests that the inhibitory action of FS390 on neurotransmitter release appeared after the influx of Ca2+.
Adenosine Triphosphate ; Animals ; Exocytosis ; Ionomycin ; Mass Screening* ; Neurons ; Neurotransmitter Agents* ; Norepinephrine ; PC12 Cells* ; Rats ; Scintillation Counting ; Streptomyces

PURPOSE: 14C-urea breath test (UBT) is a non-invasive and reliable method for the diagnosis of Helicobacter pylori (HP) infection. In this study, we evaluated the diagnostic performance of a new and rapid 14C-UBT (Heliprobe method), which was equipped with Geiger-Muller counter and compared the results with those obtained by using the conventional method. MATERIALS AND METHODS: Forty-nine patients with dyspepsia underwent gastroduodenoscopy and 14C-UBT. A 37 KBq 14C-urea capsule was administered to patients and breath samples were collected. In Heliprobe method, patients exhaled into a Heliprobe BreathCard for 10 min. And then the activities of the BreathCard were countered using Heliprobe analyzer. In the conventional method, results were countered using liquid scintillation counter. During gastroduodenoscopy, 18 of 49 patients were underwent biopsies. According to these histologic results, we evaluated the diagnostic performance of two different methods and compared them. Also we evaluated the concordant and disconcordant rates between them. RESULTS: In all 49 patients, concordant rate of both conventional and Heliprobe methods was 98% (48/49) and the discordant rate was 2% (1/49). Thirteen of 18 patients to whom biopsies were applied, were found to be HP positive on histologic results. And both Heliprobe method and conventional method classified 13 of 13 HP-positive patients and 5 of 5 HP-negative patients correctly (sensitivity 100%, specificity 100%, accuracy 100%). CONCLUSION: The Heliprobe method demonstrated the same diagnostic performance compared with the conventional method and was a simpler and more rapid technique.
Biopsy ; Breath Tests* ; Diagnosis* ; Dyspepsia ; Helicobacter pylori* ; Helicobacter* ; Humans ; Scintillation Counting ; Sensitivity and Specificity

PURPOSE: Hippocampus contains interneurons where neuropeptide Y is located, but the connectivity of these has not been well studied. Neuropeptide Y may influence the serotonergic nervous system through the interneurons. Serotonergic nerve fibers pass through nearly all areas of the hippocampus. We investigate the effects of Neuropeptide Y on serotonin release from rat hippocampal slices for the better understanding of the effects of neuropeptide Y at the hippocampus. MATERIALS AND METHODS: The hippocampus was obtained from the male rat brain and sliced. The slices were incubated in a buffer containing 0.1 mM [3H]5-hydroxytryptamine (5-HT) for uptake. The release of 5-HT into the buffer during each 10 min period was measured and the radioactivities in each buffer and the tissue were counted using liquid scintillation counter and the results were expressed as a percentage of the total activity. After 50 min from the initiation, neuropeptide Y were administered at 6th and 7th 10 min period, respectively. The changes of 5-HT release were expressed as percent values compared to the 5th 10 min period. RESULTS: A steady release of 5-HT was observed up to 100 min after the rapid release during the first 40 min. The 5-HT release during 10 and 20 min of neuropeptide Y (10 6 M) treatment showed no significant change. CONCLUSIONS: The release of 5-HT was not changed by neuropeptide Y and this results suggest that neuropeptide Y does not influence the serotonergic nervous system through the interneurons.
Animals ; Brain ; Hippocampus* ; Humans ; Interneurons ; Male ; Nerve Fibers ; Nervous System ; Neuropeptide Y* ; Neuropeptides* ; Radioactivity ; Rats* ; Scintillation Counting ; Serotonin*

PURPOSE: The C-14 urea breath test (C-14 UBT) is the most specific noninvasive method to detect Helicobacter (H) pylori infection. We investigated if the C-14 UBT can reflect the presence and degree of H. pylori detected by gastroduodenoscopic biopsies (GBx). MATERIALS AND METHODS: One hundred fifty patients (M:F=83:67, age 48.6+/-11.2 yrs) underwent C-14 UBT, rapid urease test (CLO test) and GBx on the same day. For the C-14 UBT, a single breath sample was collected at 10 minutes after ingestion of C-14 urea (137 KBq) capsule and counting was done in a liquid scintillation counter for 1 minute, and the results were classified as positive ( 200 dpm), intermediate (50~199 dpm) or negative (<50 dpm). The results of CLO tests were classified as positive or negative according to color change. The results of GBx on giemsa stain were graded 0 (normal) to 4 (diffuse) according to the distribution of H. pylori by the Wyatt method. We compared C-14 UBT results with GBx grade as a gold standard. RESULTS: In the assessment of the presence of H. pylori infection, the C-14 UBT global performance yielded sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV) and accuracy of 92.5%, 88.4%, 97.1%, 88.4% and 91.3%, respectively. However, the CLO test had sensitivity, specificity, PPV, NPV and accuracy of 83.2%, 81.4%, 91.8%, 81.4% and 82.7%, respectively. The quantitative values of the C-14 UBT were 45+/-27 dpm in grade 0, 707+/-584 dpm in grade 1, 1558+/-584 dpm in grade 2, 1851+/-604 dpm in grade 3, and 2719+/-892 dpm in grade 4. A significant correlation (r=0.848, p<0.01) was found between C-14 UBT and the grade of distribution of H. pylori infection on GBx with giemsa stain. CONCLUSION: We conclude that the C-14 UBT is a highly accurate, simple and noninvasive method for the diagnosis of ongoing H. pylori infection and reflects the degree of bacterial distribution.
Azure Stains ; Biopsy ; Breath Tests* ; Diagnosis ; Eating ; Helicobacter pylori* ; Helicobacter* ; Humans ; Scintillation Counting ; Sensitivity and Specificity ; Urea* ; Urease

Scintillation proximity assay (SPA) is a unique type of radioimmunoassay and makes it possible to use radioisotopes for monitoring binding reactions continuously without separation procedure. Microbeads containing a fluorophor are covalently linked to antibody or receptor. When a radiolabeled antigen or ligand is added it binds to the beads and the emitted short range electrons, excite the fluorophor in the beads. The light emitted can be measured in a scintillation counter. 3H or 125I has been used for SPA. The sensitivities achieved with SPA are comparable to the sensitivities of other procedures. SPA is applicable to immunology, receptor binding, monitoring interactions of biomolecules and study for the kinetics of interaction between receptors and ligands.
Allergy and Immunology ; Kinetics ; Ligands ; Microspheres ; Radioimmunoassay ; Radioisotopes ; Scintillation Counting

BACKGROUNDS: Polyamines are known to be essential for cell growth and differentiation. Recently, possible roles of the polyamine in signal transduction as neurotransmitter, modulator, or second messenger are suggested in many studies. Furthermore, it is widely studied that possible roles of polyamine are involved in the action of hormone. Thus, it was to investigate the effect of polyamines in the cell proliferation and secretion of GH from the GH cells. METHODS: Cells(5*10 cells/mL) were incubated for 3 days in DMEM containing test drugs and labeled with 20pCi/mL of [S]-methionine for 2 hr. Proteins secreted into the medium were separated by 13% SDS-gel electrophoresis, then autoradiography was performed to identify radiolabeled proteins. [S]-methionine labelled GH was identified by radioimmuno-precipitation. Total protein synthesis was determined from the radioactivity of the cell homogenate by liquid scintillation counter. The intracellular polyamine content was determined by HPLC. RESULTS: Externally added polyamines(putrescine, spermidine, spermine) induced cell proliferation in a dose-dependent manner at proper concentrations, specifically 50pM putrescine increased GH secretion, DFMO or MGBG, which is polyamine biosynthetic inhibitor, inhibited GH secretion in a dose-dependent fashion, In the cells treated with 20mM or 0.01mM MGBG, total protein synthesis were decreased only to 90 or 76% of the control levels and cell proliferation was also slightly inhibited. However the secretion of GH was severely blocked to 37% or 35% of the control. Hydrocortisone at 5 pM stimulated the secretion of GH to 153% of basal secretion, also doubled intracellular putrescine content. CONCLUSION: The present data show that externally added polyamines induced cell proliferation and GH secretion. Also, extemally added putrescine stimulated GH secretion significantly. GH secretion was inhibited by polyamine metabolic inhibitor in a dose-dependent manner and polyamine metabolic inhibitors, at proper concentrations, specifically blocked GH secretion without any significant influence on the total protein synthesis. The above results imply the involvement of polyamine in GH secretion.
Autoradiography ; Cell Proliferation ; Chromatography, High Pressure Liquid ; Electrophoresis ; Growth Hormone ; Hydrocortisone ; Mitoguazone ; Neurotransmitter Agents ; Polyamines ; Putrescine ; Radioactivity ; Scintillation Counting ; Second Messenger Systems ; Signal Transduction ; Spermidine

5-hydroxytryptamine (5-HT) containing pathways exert a inhibitory or facilitate effect on copulation. The administration of gamma aminobutyric acid (GABA) receptor agonists to the central nervous system in rats inhibits male copulatory behavior, whereas the administration of antagonists facilitates corpulatory behavior. GABA may influence the erection through dopaminergic pathway. But few about the effect of GABA on serotoninergic neuron are known. Raphe nuclei give rise to the major serotonergic innervation to the hippocampal formation and the GABAergic interneurons are located at the hippocampal formation also. This study was performed to investigate the influence of GABA, inhibitory interneuron, on the 5-HT release from rat hippocampal slices to understand the connection of the serotonergic neurons to GABAergic interneurons. The hippocampus was obtained from the male rat brain and sliced to a 400 Im thickness. After 30 minutes preincubation in the normal buffer, the slices were incubated for 20 minutes in a buffer containing 0.1 microM [3H]5-HT for uptake, and washed. After administration of GABA, the release of [3H] 5-HT into the buffer was measured and the radioactivities in each buffer and the tissue were counted using liquid scintillation counter and the results were expressed as a percentage of the total activity. Spontaneous release of [3H] 5-HT from the rat hippocampal slices decreased rapidly during the first 40 minutes of incubation. Through the rapid release of [3H] 5-HT, a steady state of [3H] 5-HT release was obtained from the 50 minutes of incubation. The value of released [3H] 5-HT after 50 minutes was expressed as percent of the value at 50 minutes. After administration of GABA (10(-4)M), the values (mean +/- SE) of released (3H)5-HT were 97.3 +/- 3.8% at 60 minutes and 91.2 +/- 3.4% at 70 minutes. The values of control group were 96.6 +/- 1.9% at 60 minutes and 89.6 +/- 2.3% at 70 minutes. There were no changes in the release of (3H)5-HT after administration of GABA. These results suggest that there are few connections between GABA and serotoninergic neurons and GABA does not influence the release of 5-HT in rat hippocampus.
Animals ; Brain ; Central Nervous System ; Copulation ; gamma-Aminobutyric Acid* ; Hippocampus ; Humans ; Interneurons ; Male ; Radioactivity ; Raphe Nuclei ; Rats ; Scintillation Counting ; Serotonergic Neurons ; Serotonin*

PURPOSE: Phospholipase D (PLD) catalyzes the hydrolysis of phosphatidyl choline to phosphatidic acid (PA) and choline. Recently, PLD has been drawing much attentions and considered to be associated with cancer process since it is involved in cellular signal transduction. In this experiment, oleate-PLD activities were measured in various tissues of the living rats after whole body irradiation. MATERIAL AND METHODS: The reaction mixture for the PLD assay contained 0.1microCi 1,2-di[1-14C]palmitoyl phosphatidylcholine, 0.5mM phosphatidylcholine, 5mM sodium oleate, 0.2% taurodeoxycholate, 50mM HEPES buffer (pH 6.5), 10mM CaCl2, and 25mM KF. phosphatidic acid, the reaction product, was separated by TLC and its radioactivity was measured with a scintillation counter. The whole body irradiation was given to the female Wistar rats via Cobalt 60 Teletherapy with field size of 10cm x 10cm and an exposure of 2.7Gy per minute to the total doses of 10Gy and 25Gy. RESULTS: Among the tissues examined, PLD activity in lung was the highest one and was followed by kidney, skeletal muscle, brain, spleen, bone marrow, thymus, and liver. Upon irradiation, alteration of PLD activity was observed in thymus, spleen, lung, and bone marrow. Especially PLD activities of the spleen and thymus revealed the highest sensitivity toward gamma-ray with more than two times amplification in their activities. In contrast, the PLD activity of bone marrow appears to be reduced to nearly 30%. Irradiation effect was hardly detected in liver which showed the lowest PLD activity. CONCLUSION: The PLD activities affected most sensitively by the whole-body irradiation seem to be associated with organs involved in immunity and hematopoiesis. This observation strongly indicates that the PLD is closely related to the physiological function of these organs. Furthermore, radiation stress could offer an important means to explore the phenomena covering from cell proliferation to cell death on these organs.
Animals ; Attention ; Bone Marrow ; Brain ; Cell Death ; Cell Proliferation ; Choline ; Cobalt ; Female ; Hematopoiesis ; HEPES ; Humans ; Hydrolysis ; Kidney ; Liver ; Lung ; Muscle, Skeletal ; Oleic Acid ; Phosphatidic Acids ; Phosphatidylcholines ; Phospholipase D* ; Phospholipases* ; Radiation Effects ; Radioactivity ; Rats* ; Rats, Wistar ; Scintillation Counting ; Signal Transduction ; Sodium ; Spleen ; Taurodeoxycholic Acid ; Thymus Gland ; Whole-Body Irradiation