To investigate the influence of phenobarbital sodium on the action of morphine and on the diurnal rhythms of both opiate receptor binding and ${\beta}-endorphin$ contents, the amount of specifically bound $(^3H)$-morphine and immunoreactive ${\beta}-endorphin$ were measured in the midbrain of phenobarbital-treated rats at 4h intervals in a day. Rats were housed and adapted to a controlled cycle of either 12 h light-12 h dark or 24 h constant dark. After 3 weeks of adaptation, 0.5 ml of physiological saline or phenobarbital sodium (20mg/kg/day, i.p.) were administered twice a day for 2 weeks. Highly significant diurnal rhythms of opiate receptor binding and ${\beta}-endorphin$ were present in rat midbrain. In control group, the peak of maximum $(^3H)$-morphine binding was observed at 22:00 h, whereas the peak of ${\beta}-endorphin$ content was found at 06:00 h. Even in the absence of time cues these diurnal rhythms persisted, but they were highly modified with respect to the wave form as well as differences in the timing of peak and nadir. In the phenobarbital-treated group, these diurnal rhythms were also modified in shape, phase and amplitude, as well as in timing of peak and nadir. In this group, 24 h mean of opiate receptor binding was significantly decreased, while the 24 h mean level of ${\beta}-endorphin$ content was highly increased. However, Kd values in all experimental groups did not change. This indicates that differences in binding were not due to changes in the affinity, but in the number of binding sites. Statistical analysis of regression line indicates that changes of receptor binding were closely correlated with the changes of ${\beta}-endorphin$ content. These results suggest that phenobarbital may influence the action of morphine by changing the number of opiate receptors and that the modification of diurnal rhythm of opiate receptor by the agent is possibly due to changes of ${\beta}-endorphin$ content.
To detect ischemic tissue in experimental model of cerebral ischemia made by middle cerebral artery(MCA)-occlusion, we acquired triple image of $^{99m}Tc$-glucarate, [$^{18}F$]fluoro-deoxyglucose (FDG), and 2,3,5- triphenyltetrazolium (TTC) staining. We made cerebral infarction either with reperfusion (after occlusion of 2 hours) or without reperfusion in 10 Sprague-Dawley rats by inserting thread to MCA through internal carotid artery. After 22 hours, we injected 740 MBq of $^{99m}Tc$-glucarate and 55.5 MBq of [$^{18}F$]FDG through tail vein. Each 1 mm slice of rat brains was frozen and exposed to imaging plate for 20 minutes in freezer to get an [$^{18}F$]FDG image. After 20 hours enough to fade radioactivity of [$^{18}F$]FDG, the slices were again imaged by BAS1500 for $^{99m}Tc$-glucarate uptake. Finally, these brain tissues were stained with TTC. Semi-quantitative visual analysis was done by grading 0 to 3 points according to the degree of uptakes($^{99m}Tc$-glucarate) and decreased uptakes([$^{18}F$]FDG and TTC). Ten rats survived with neurologic symptoms. TTC staining confirmed the development of infarction. The size of the infarction was relatively larger in the group without reperfusion. [$^{18}F$]FDG images were similar to TTC-stained images. However, we found regions with intermediate uptake which were not stained with TTC. We found regions with intermediate [$^{18}F$]FDG uptake where TTC staining was normal. $^{99m}Tc$-glucarate uptake was round only in TTC non-stained region. In the TTC stained regions, there were no uptake of $^{99m}Tc$-glucarate. We could not find clear relation between $^{99m}Tc$-glucarate uptake with [$^{18}F$]FDG uptake. This was partly because percent uptake of $^{99m}Tc$-glucarate was so small (less than 1 percent of injected dose) and because there were quite heterogeneity of patterns of [$^{18}F$]FDG uptake and TTC. With these findings, we could conclude that $^{99m}Tc$-glucarate were taken up only in part of ischemic tissues which were proven to be nonviable. The establishment of MCA-occluded rat model with or without reperfusion and triple imaging for $^{99m}Tc,\;^{18}F$ and TTC helped the characterization of $^{99m}Tc$-glucarate uptakes. Further work is needed to clarify the meaning or diversities or [$^{18}F$]FDG and TTC and their relation with $^{99m}Tc$-glucarate.
Kim, Un-Sung;Lee, Cherl-Ho;Kim, Seong-Jo;Lee, Joo-Don;Moon, Kwang-Hyun;Baek, Seung-Hwa
Korean Journal of Food Science and Technology
/
v.27
no.4
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pp.555-563
/
1995
This study was performed to investigate the effect of Aloe arborescens on the cadmium toxicity in rats. Thirty male Sprague-Dawley strains were divided into five groups consisting of a control group, a cadmium treatment group and 3 aloe(0.5%, 0.75%, 1%) treatment groups and observed for 9 weeks. The weight increment of the cadmium and 0.75% aloe group was higher than that of the cadmium treatment group(p<0.01). The food intake did not show the consistency rule among the experimental groups and the decrement tendency of food intake affected by cadmium feeding group. The decrement tendency of water intake affected by cadmium appeared to be suppressed by aloe treatment, especially cadmium and 0.75% aloe treatment group showed the remarkable increment of water intake. The diet efficiency of the control group was the highest among the experimental groups and that of cadmium and 0.75% aloe group was higher than other aloe treatment groups. The weight of each organ did not show consistency among the experimental groups but only the testicle of cadmium and 0.75% aloe treatment group was heavier than that of the control group. The cadmium accumulation was high in order of kidney>liver>spleen>heart>lung>testicle>brain. The cadmium content of the cadmium treatment group was more than that of cadmium and 0.5% aloe group, cadmium and 0.75% aloe group, cadmium and 1% aloe group. The cadmium content of cadmium and 0.75% aloe group was the lowest among other aloe treatment groups. Therefore, cadmium and 0.75% aloe is the most recommendable aloe treatment to eliminate the cadmium accumulated in organ.
Journal of the Korean Society of Food Science and Nutrition
/
v.41
no.4
/
pp.471-479
/
2012
This study was conducted to investigate the effects of various levels of chitosan oligomer (CO) molecular weight on the disorders of lipid metabolism and immune-related factors induced by 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), that is a endocrine disrupter, using adult male rats (SD) for 3 weeks. These 40 animals were divided into five groups. Three kinds of CO were used by molecular weight (MW) (less than 1000, 1000~3000, and 5000~10000) and added 4% to basal diets respectively. TCDD (40 ${\mu}g$/kg B.W) was intraperitoneally injected into rats at the beginning of the experiment. The relative weights of the livers were increased in all rats treated with TCDD, and the brain and testis weights were increased in all CO diet groups, compared to the control and TCDD groups. The levels of white blood cells (WBC) and red blood cells (RBC), hemoglobin, hematocrits (HCT), and platelets were significantly lowered by treating TCDD. By the way, RBC and HCT tended to recover by CO diets. The elevation of serum total and HDL cholesterol levels induced by TCDD treatment was significantly reduced by CO (5000~10000 MW) diets. The apparent increasing of the total lipid, cholesterol, and triglyceride levels of rat livers induced by TCDD was tended to be suppressed in those fed CO diets. Especially, diets with less than 1000 MW significantly diminished liver triglycerides. The levels of serum immunoglobulin (Ig) A, IgG1 and IgM were significantly high in rats fed CO (5000~10000 MW) diets. The decreasing levels of IgE by treatment with TCDD tended to recover all the CO diet groups to the level of control group. In histochemical observation, the fat droplets and apoptosis of liver due to TCDD treatment were markedly alleviated in all CO diet groups. These results indicated that CO, though not regular according to molecular weight, can exert improving effects on lipid accumulation, hepatocytic disorders, abnormal blood cells, and some immunoglobulins induced by TCDD.
Purpose: Lingual nerve (LN) damage may be caused by either tumor resection or injury such as wisdom tooth extraction, Although autologous nerve graft is sometimes used to repair the damaged nerve, it has the disadvantage of necessity of another operation for nerve harvesting. Moreover, the results of nerve grafting is not satisfactory. The nerve growth factor (NGF) is well-known to play a critical role in peripheral nerve regeneration and its local delivery to the injured nerve has been continuously tried to enhance nerve regeneration. However, its application has limitations like repeated administration due to short half life of 30 minutes and an in vivo delivery model must allow for direct and local delivery. The aim of this study was to construct a well-functioning $rhNGF-{\beta}$ adenovirus for the ultimate development of improved method to promote peripheral nerve regeneration with enhanced and extended secretion of hNGF from the injured nerve by injecting $rhNGF-{\beta}$ gene directly into crush-injured LN in rat model. Materials and Methods: $hNGF-{\beta}$ gene was prepared from fetal brain cDNA library and cloned into E1/E3 deleted adenoviral vector which contains green fluorescence protein (GFP) gene as a reporter. After large scale production and purification of $rhNGF-{\beta}$ adenovirus, transfection efficiency and its expression at various cells (primary cultured Schwann cells, HEK293 cells, Schwann cell lines, NIH3T3 and CRH cells) were evaluated by fluorescent microscopy, RT-PCR, ELISA, immunocytochemistry. Furthermore, the function of rhNGF-beta, which was secreted from various cells infected with $rhNGF-{\beta}$ adenovirus, was evaluated using neuritogenesis of PC-12 cells. For in vivo evaluation of efficacy of $rhNGF-{\beta}$ adenovirus, the LNs of 8-week old rats were exposed and crush-injured with a small hemostat for 10 seconds. After the injury, $rhNGF-{\beta}$ adenovirus($2{\mu}l,\;1.5{\times}10^{11}pfu$) or saline was administered into the crushed site in the experimental (n=24) and the control group (n=24), respectively. Sham operation of another group of rats (n=9) was performed without administration of either saline or adenovirus. The taste recovery and the change of fungiform papilla were studied at 1, 2, 3 and 4 weeks. Each of the 6 animals was tested with different solutions (0.1M NaCl, 0.1M sucrose, 0.01M QHCl, or 0.01M HCl) by two-bottle test paradigm and the number of papilla was counted using SEM picture of tongue dorsum. LN was explored at the same interval as taste study and evaluated electro-physiologically (peak voltage and nerve conduction velocity) and histomorphometrically (axon count, myelin thickness). Results: The recombinant adenovirus vector carrying $rhNGF-{\beta}$ was constructed and confirmed by restriction endonuclease analysis and DNA sequence analysis. GFP expression was observed in 90% of $rhNGF-{\beta}$ adenovirus infected cells compared with uninfected cells. Total mRNA isolated from $rhNGF-{\beta}$ adenovirus infected cells showed strong RT-PCR band, however uninfected or LacZ recombinant adenovirus infected cells did not. NGF quantification by ELISA showed a maximal release of $18865.4{\pm}310.9pg/ml$ NGF at the 4th day and stably continued till 14 days by $rhNGF-{\beta}$ adenovirus infected Schwann cells. PC-12 cells exposed to media with $rhNGF-{\beta}$ adenovirus infected Schwann cell revealed at the same level of neurite-extension as the commercial NGF did. $rhNGF-{\beta}$ adenovirus injected experimental groups in comparison to the control group exhibited different taste preference ratio. Salty, sweet and sour taste preference ratio were significantly different after 2 weeks from the beginning of the experiment, which were similar to the sham group, but not to the control group.
The effects of medetomidine on the degree of analgesia and sedation in rats were evaluated. The rats were randomly divided into six groups: saline, 1 mL/kg (group 'Saline'); butorphanol, 2.0 mg/kg; medetomidine, 0.2, 0.4, 0.8 or 1.6 mg/kg (group 'MED0.2', 'MED0.4', 'MED0.8' and 'MED1.6', respectively). The degree of analgesia was measured in the $50^{\circ}C$ hot-water tail-flick latency test, and the degree of sedation was evaluated using the numerical sedation score (NSS) and the righting reflex. All doses of medetomidine, except MED0.2, significantly increased the analgesic effect compared to the Saline group. Variables in the MED0.4 and MED0.8 groups, but not in the MED1.6 group, were significantly increased compared to those in the MED0.2 group. However, analgesia with all doses of medetomidine was not significantly different compared to that with butorphanol. Saline and butorphanol treatments did not induce sedation and loss of righting reflex during the recording period. NSS in the MED0.4, MED0.8 and MED1.6 groups were significantly higher than that in the MED0.2 group. NSS in the MED0.8 and MED1.6 groups were not significantly different from that in the MED0.4 group. The latency to loss of righting reflex in the MED0.8 and MED1.6 groups decreased significantly compared to that in the MED0.2 group. Thus, 0.4 and 0.8 mg/kg of medetomidine provided not only reliable analgesia but also sedation to rats. In conclusion, 0.4 to 0.8 mg/kg medetomidine could be a useful chemical restraint method in rats.
Purpose : This study aimed to investigate whether exogenous thyroxine($T_4$) treatment to alcohol-fed dams would ameliorate the detrimental effects of alcohol on the postnatal development of neuropeptide-Y(NPY)-containing neurons of the cerebral cortex and hippocampus of the offspring. Methods : Time-pregnant rats were divided into three groups. An alcohol-fed group A received 35 calories of liquid alcohol diet daily from gestation day 6; control group B was fed a liquid diet in which dextrin replaced alcohol isocalorically; and alcohol+$T_4$ group C received 35 calories of liquid alcohol diet and exogenous thyroxine subcutaneously. The features of the growth and maturation of rat brain tissue were observed at 0, 7, 14, 21 and 28 postnatal days via immunohistochemistry. Results : Group C showed prominent NPY immunoreactivity in the cerebral cortex compared to group A and B at P7. In group C, NPY-containing neurons were widely distributed in the all layers of cerebral cortex after P14. Also, numerical decreases of NPY-containing neuron were not found according to increasing age in group C. A decrease of NPY-containing neurons, however, was clearly observed in group A compared to group C at P28. In the hippocampus, similar patterns appeared in groups B and C after P7. Especially, in groups B and C, NPY-containing fibers formed plexus in the cerebral cortex and hippocampus at P14. Conclusion : These results suggest that the increase of NPY synthesis caused by maternal administration of exogenous thyroxine may convalesce fetal alcohol effects, one of the effects of the dysthyroid state following maternal alcohol abuse.
Purpose : We aimed to investigate the efficacy of and functional recovery after intracerebral transplantation of different doses of mouse mesenchymal stem cells (mMSCs) in immature rat brain with hypoxic-ischemic encephalopathy (HIE). Methods : Postnatal 7-days-old Sprague-Dawley rats, which had undergone unilateral HI operation, were given stereotaxic intracerebral injections of either vehicle or mMSCs and then tested for locomotory activity in the 2nd, 4th, 6th, and 8th week of the stem cell injection. In the 8th week, Morris water maze test was performed to evaluate the learning and memory dysfunction for a week. Results : In the open field test, no differences were observed in the total distance/the total duration (F=0.412, P=0.745) among the 4 study groups. In the invisible-platform Morris water maze test, significant differences were observed in escape latency (F=380.319, P<0.01) among the 4 groups. The escape latency in the control group significantly differed from that in the high-dose mMSC and/or sham group on training days 2-5 (Scheffe's test, P<0.05) and became prominent with time progression (F=6.034, P<0.01). In spatial probe trial and visible-platform Morris water maze test, no significant improvement was observed in the rats that had undergone transplantation. Conclusion : Although the rats that received a high dose of mMSCs showed significant recovery in the learning-related behavioral test only, our data support that mMSCs may be used as a valuable source to improve outcome in HIE. Further study is necessary to identify the optimal dose that shows maximal efficacy for HIE treatment.
Synapses are contact points where one neuron communicates with another. The morphological change of synapses under various physiological or pathological conditions has long been hypothesized to modify their functional properties. 3-dimensional (3-D) reconstruction of synapses with serial ultrathin sections has contributed to the understanding of ultrastructural dimensions and compositions of synapses. The 3-D reconstruction procedures, however, require a great amount of expertise as well as include prohibitively timeconsuming processes. Here, we introduce efficient 3-D reconstruction technique using high-voltage electron microscopy (HVEM). Primarily, we established an optimal section thickness and staining condition to observe synaptic structures in detail under HVEM. The result showed that synaptic profiles were preserved at the section thickness of 250 nm without the overlapping of synaptic ultrastructures. An increase in the reaction time of en bloc staining was most efficient to enhance contrast than the extension of postembedding staining or the addition of uranyl acetate during dehydration. Then, 3-D reconstruction of parallel fiber-Purkinje cell synapses in the rat cerebellum was carried out with serial HVEM images and reconstruction software. The images were aligned and the contours of synapses were outlined on each section. 3-D synapses were finally extracted from the section files by grouping all the synaptic contours. The reconstructed synapse model clearly demonstrated the configuration of pre and postsynaptic components. These results suggest that 3-D reconstruction of synapses using HVEM is much efficient and suitable for massive quantitative studies on synaptic connectivity than conventional TEM approach using numerous ultrathin sections.
Journal of the Korean Society of Food Science and Nutrition
/
v.31
no.6
/
pp.1084-1091
/
2002
Mortierella alpina, a common soil fungus, is the most efficient organism for production of production acid presently known. Since arachidonic acid are important in human brain and retina development, it was undertaken the growing effect containing diet as a food ingredient. Arachidonic acid rich oil derived from Mortierella alpina, was subjected to a program of studies to establish for use in diet supplement. This study was compared the growth and learning effect of fungal oil rich in arachidonic acid by incorporated into diets ad libitum. Sprague-Dawley rats received experimental diets 5 groups (standard AIN 93 based control with beef tallow, extract oil 8%, and 4%, and Mortierella alpina in diet 10% and 20%) over all experiment duration (pre-mating, mating, gestation, lactation, and after weaning 4 weeks). Pups born during this period consumed same diets after wean for 4 weeks. There was no statistical significance of diet effects in reproductive performance and fertility from birth to weaning. But the groups of Mortierella alpine diet were lower of weight gain and diet intake after weaning. The serum lipids were significantly different with diet groups, higher TG in LO (oil 4%) group of dams, and higher total cholesterol in LF (M. alpina 10%) of pups, although serum albumin content was not significantly different in diet group. The spent-time and memory effect within 4 weeks of T-Morris water maze pass test in dam and 7-week- age pups did not differ in diet groups. On the count of backing error in weaning period of pups was lower in HO(extracted oil 8%) group. In the group of 10% and 20% Mortierella alpina diet, DNA content was lower in brain with lower body weight, but liver DNA relative to body weight was higher than control. Further correlation analyses would be needed DNA and arachidonic acid intakes, with Mortierella alpina diet digestion rate.
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