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ABSTRACT: The development of automated detection systems for animal behaviors is increasing in value in terms of saving time, objective analysis and reducing the need for well-trained experimenters. SCLABA(®) (Noveltec Inc., Kobe, Japan) is a commercially available analysis system originally developed for analyzing scratching behaviors in rodents, based on distances between points in videotaped images. Here, we used this software to automate analysis of abdominal licking behavior associated with visceral pain in mice.
Yellow and green spots were applied to the snout and the lower abdominal region of mice respectively to provide reference points for automated analysis of video recordings. Abdominal licking behavior after intracolonic administration of 0.3% capsaicin solution as a measure of visceral pain was determined based on changes in the inter-spot distance.
A distance threshold between the colored spots was chosen based on manual measurements showing that 99% of minimal distances were below this threshold. Using this threshold, the number of licks determined by the automated analysis significantly and positively correlated with that determined by manual observation (R(2)=0.95 and p<0.001). The neurokinin-1 receptor antagonist GR205171A dose-dependently inhibited capsaicin-induced licking detected by automated analysis.
We demonstrated that visceral pain-related licking behaviors after intracolonic capsaicin treatment can be automatically detected by applying commercially available image analysis software. This automated experimental system is very efficient and useful to evaluate antinociceptive effect of a test compound on visceral pain.
Journal of pharmacological and toxicological methods 03/2011; 64(2):119-23. · 2.32 Impact Factor
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ABSTRACT: Marble burying and forced swimming behavior are widely used and sensitive tests for identifying clinically effective antidepressant drugs, although the underlying neurobiology of these behaviors is not fully elucidated.
The objective of this study was to determine the relationship between the behavioral effects of antidepressant drugs and their ability to modulate extracellular neurotransmitter levels in the prefrontal cortex.
The effects of fluoxetine, fluvoxamine, citalopram, imipramine, and desipramine (0 to 60 mg/kg by oral gavage, except fluoxetine at 0 to 40 mg/kg) were studied independently in CD-1 mice in the marble-burying task, forced swim task and on extracellular concentrations of serotonin, norepinephrine, and dopamine in the prefrontal cortex by freely moving microdialysis.
Fluvoxamine, fluoxetine, and citalopram all suppressed marble-burying behavior, but produced no change in immobility time in the forced swim test. In contrast, imipramine and desipramine suppressed both marble-burying behavior and increased swimming time in the forced swim test, although desipramine mildly suppressed locomotor activity at the maximal dose. Fluvoxamine, fluoxetine, and citalopram all increased extracellular levels of cortical serotonin. Desipramine and imipramine increased extracellular dopamine levels. Fluoxetine, desipramine, and imipramine increased extracellular norepinephrine levels. Correlational analysis revealed a positive correlation between efficacy of drugs in the forced swim test and cortical extracellular dopamine levels, whereas a positive correlation was found between efficacy in the marble-burying test and extracellular serotonin levels.
Although marble burying and forced swimming behavior have strong predictive validity in tests of antidepressant action, each assay appears to be underpinned by entirely different neurochemical systems.
Psychopharmacologia 06/2008; 197(4):567-80. · 4.08 Impact Factor
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ABSTRACT: T cells play an important role in initiating autoimmune responses and maintaining synovial inflammation in rheumatoid arthritis. Although, anti-type II collagen antibody-induced arthritis (CAIA) is generally believed to be a T cell- and B cell-independent model, the detailed pathogenesis of CAIA remains unclear. In the present study, to elucidate the contribution of T cells to the pathogenesis of CAIA, we evaluated the effects of CTLA4 Ig and cyclosporin (CsA). Arthritis was induced in mice by intravenous injection of anti-type II collagen antibody followed by intraperitoneal injection of lipopolysaccharide. CTLA4 Ig was intraperitoneally administered and CsA was subcutaneously administered; then the severity of arthritis was evaluated by scoring the edema and erythema of paws and by measuring hind paw thickness. Paw samples were collected 12 days after the antibody injection, and the mRNA expression levels were analyzed by real-time quantitative polymerase chain reaction. Administration of CTLA4 Ig ameliorated the increases in arthritic score and paw thickness in the later phase, but not in the early phase of arthritis. CsA suppressed the increases in arthritic score and paw thickness in both the early and later phases of arthritis. CTLA4 Ig and CsA suppressed mRNA up-regulation of T-cell markers, CD3 and CD25, and immune response-related mediators, IFN-gamma and IL-12. They also suppressed the up-regulation of macrophage marker, F4/80, and proinflammatory cytokines, TNF-alpha, IL-1beta and IL-6. The results provide direct evidence that arthritis in this model is T-cell activation dependent.
International Immunopharmacology 11/2007; 7(10):1360-8. · 2.38 Impact Factor
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ABSTRACT: Reactive Oxygen Spiecies (ROS) such as superoxide anion radical (.O(2)(-)) act as signals for the activation of stress-response and defense pathways. However, excess ROS generated by perturbing .O(2)(-) homeostasis stimulated many environmental stress, including intense light, drought, temperature stress, herbicides, induce high radical toxicity. Consequently, quantitative analysis of .O(2)(-) is a subject of intense research, since most of ROS are derived from .O(2)(-). Iron meso-tetrakis(3-thienyl)porphyrin complexes were electropolymerized onto a Au wire electrode. The modified Au electrode were applied to .O(2)(-) sensor to detect catalytic oxidation current of .O(2)(-) which was generated as an intermediate during the oxidation of xanthine by catalystic XOD. It was revealed that the sensor was quantitative to measure .O(2)(-). The modified Au electrode were applied to measure oxidation current of .O(2)(-) in mung beans under environmental stress condition. Plants were grown in atmosphere, 25 degrees C and in black darkness. The other plants were exposed to oxygen excess. The oxidation current of .O(2)(-) were increased plants were grown by high-oxygen environment compared to plants were grown at atmosphere. This experiment was indicated that environmental stress such as hyperoxia induced excess .O(2)(-) and Au wire sensor using iron porphyrin complexes is capable of .O(2)(-) detection in plants under environmental stresses.
Journal of oleo science 02/2007; 56(2):81-6. · 1.42 Impact Factor
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ABSTRACT: Oxidative polymerization of bromoiron(III) meso-tetrakis(3-thienyl)porphyrin gave a novel polymeric porphyrin complex randomly crosslinked at the 2,5-positions of the peripheral thienyl groups. The electrical semiconductivity of ca. 10−5 S/cm after I2 doping indicated that the polymer had a π-conjugated structure with a moderate delocalization of π electrons over the thienylporphyrin units. PM3 calculations for free-base models revealed that HOCO (the highest occupied crystal orbital) band width was reduced by introduction of the porphyrin moieties into the thienylene backbone and yet low HOCO-LUCO (the lowest unoccupied crystal orbital) gap was maintained, which accounted for the relatively low electrical conductivity of the porphyrin polymer. The modified electrode prepared by electropolymerization was redox-active due to the presence of iron(II/III) couple and the semiconductivity of the film, which served as a novel non-enzymatic electrochemical sensor for superoxide anion radical based on the facile electrocatalytic oxidation of the superoxide. Copyright © 2005 John Wiley & Sons, Ltd.
Polymers for Advanced Technologies 06/2005; 16(8):616 - 621. · 2.01 Impact Factor
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ABSTRACT: A needle-type electrochemical sensor for the facile detection of superoxide anion radical (O) was devised using an electrodeposited film of a polymeric porphyrin complex attached to a carbon microelectrode which was placed in a stainless steel 18G needle tube as an auxiliary electrode. The sensing element was prepared by means of electropolymerization of bromoiron(III) meso- tetra(3-thienyl)porphyrin in the presence of 1-methylimidazole in CH2Cl2 containing 100 mM tetrabutylammonium perchlorate (TBAP) as a supporting electrolyte, which gave a smooth film of the corresponding polymer. The film was electrochemically active to give a redox response near −0.05 V versus Ag/AgCl due to the iron(II/III) couple. The microsensor was applied to detect O produced by xanthine oxidation catalyzed by xanthine oxidase. The amperometric response for O was monitored at an electrode potential of 0.5 V versus the auxiliary electrode in a 10 mM phosphate buffer. The microsensor displayed a high catalytic activity for the oxidation of O and showed a linear relationship between the current and the O concentration. Axial coordination of an imidazole ligand to the iron porphyrin center enhanced selectivity for O by impeding the undesired coordination of H2O2 that resulted from the dismutation of O. Copyright © 2005 John Wiley & Sons, Ltd.
Polymers for Advanced Technologies 02/2005; 16(4):287 - 292. · 2.01 Impact Factor
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ABSTRACT: In most publications on the forced swim test studies, duration of immobility is measured during the last 4 min of the 6-min testing period as a marker for depression. However, it is not clear if 4-min span best captures antidepressant-like drug effects. In the present study, six typical antidepressants in clinical use were evaluated over time. Imipramine (60 mg/kg, PO), desipramine (60 mg/kg, PO), reboxetine (20 mg/kg, IP), bupropion (60 mg/kg, PO), fluoxetine (20 mg/kg, PO) and fluvoxamine (60 mg/kg, PO) were characterized over 1-min intervals for the 6-min testing period in order to identify more sensitive periods than the last 4-min span to detect drug effects, using the automated experimental system.
One day before the testing, wire rings were attached to the hind paws of male CD-1 mice. On the test day, after attaching a small magnet to the wire rings, each animal was placed for 6 min in a glass cylinder filled with water, which is surrounded by a coil. The duration of immobility was measured by the previously validated automated detection system, MicroAct, i.e. electrical signals generated by limb movements served as the marker for swimming behavior.
In the 1-min interval examination, it was only in 2-3-min and 3-4-min spans that all of the six antidepressants reduced the duration of immobility with statistical significance. In the 2-4-min analysis, all of the six antidepressants demonstrated statistically significant reduction in the duration of immobility, while in the 2-6-min analysis, the reduction by fluvoxamine was not statistically significant.
The detailed time course analysis of the FST in mice using the automated system revealed that the duration of immobility between 2 and 4 min is the optimum period to capture the antidepressant-like effect of test compounds.
Journal of pharmacological and toxicological methods 57(1):80-4. · 2.32 Impact Factor
