• Investigative Magnetic Resonance Imaging
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• v.9 no.1
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• pp.30-35
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• 2005

Purpose : The purpose of this study is to observe the blood oxygen level dependent (BOLD) contrast changes due to the reaction of human brain at a gustatory sense in response to a salty-taste stimulation. Materials and Methods : Twelve healthy, non-smoking, right-handed male subjects (mean age: 25.6, range: 23-28 years) participated in this salty-taste stimulus functional magnetic resonance (fMRI) study. MRI scans were performed with 1.57 GE Signa, using a multi-slice GE-EPI sequence according to a blood-oxy-gen-level dependent (BOLD) experiment paradigm. Scan parameters included matrix size $128\times128$, FOV 250 mm, TR 5000 msec, TE 60 msec, TH/GAP 5/2 mm. Sequential data acquisitions were carried out for 42 measurements with a repetition time of 5 sec for each taste-stimulus experiments. Analysis of fMRI data was carried out using SPM99 implemented in Matlab. NaCl solution $(3\%)$ was used as a salty stimulus. The task paradigm consisted of alternating rest-stimulus cycles (30-second rest, 15-second stimulus) for 210 seconds. During the stimulus period, NaCl-solution was presented to the subject's mouth through plastic tubes as a bolus of delivered every 5 sec using -processor controlled auto-syringe pump. Results : Insula, frontal opercular taste cortex, amygdala and orbitofrontal cortex (OFC) were activated by a salty-taste stimulation $(NaCl,\;3\%)$ in the fMRI experiments. And dosolateral prefrontal cortex (DLPFC) was also significantly responded to salty-taste stimuli. Activation areas of the right side hemisphere were more superior to the left side hemisphere. Conclusion : The results of this study well correspond to the fact that both insula, amygdala, OFC, DLPFC areas are established as taste cortical areas by neuronal recordings in primates. Authors found that laboratory-developed auto-syringe pump is suitable for gustatory fMRI study. Further research in this field will accelerate to inquire into the mechanism of higher order gustatory process.

• Clinical and Experimental Pediatrics
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• v.45 no.12
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• pp.1551-1558
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• 2002

Purpose : Loss of hippocampal interneurons in dentate gyrus has been reported in patients with severe temporal lobe epilepsy and in animals treated with kainic acid(KA). Interneurons contain $Ca^{2+}$- binding protein parvalbumin(PV). The effects of kainic acid on parvalbumin-immunoreactive (PV-IR) interneurons in dentate gyrus were investigated in organotypic hippocampal slice cultures. Methods : Cultured hippocampal slices from postnatal day nine C57/BL6 mice were exposed to $10{\mu}M$ KA, and were observed at 0, 8, 24, 48, 72 hours after a one hour KA exposure. Neuronal injury was determined by morphologic changes of PV-IR interneuron in dentate gyrus. Results : Transient(1 hour) exposure of hippocampal explant cultures to KA produced marked varicosities in dendrites of PV-IR interneuron in dentate gyrus and the shaft of interbeaded dendrite is often much thinner than those in control. The presence of varicosities in dendrites was reversible with KA washout. The dendrites of KA treated explants were no longer beaded at 8, 24, 48 and 72 hours after KA exposure. The number of cells in PV-IR interneurons in dentate gyrus was decreased at 0, 8 hours after exposure. But there was no significant difference in 24, 48 and 72 hours recovery group compared with control group. Conclusion : The results suggested that loss of PV-IR interneurons in dentate gyrus is transient, and is not accompanied by PV-IR interneuronal cell death.

• Journal of the Korean Society of Radiology
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• v.14 no.2
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• pp.157-168
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• 2020

Advancements in segmentation methodology has made automatic segmentation of brain structures using structural images accurate and consistent. One method of automatic segmentation, which involves registering atlas information from template space to subject space, requires a high quality atlas with accurate boundaries for consistent segmentation. The Allen Mouse Brain Atlas, which has been widely accepted as a high quality reference of the mouse brain, has been used in various segmentations and can provide accurate coordinates and boundaries of mouse brain structures for tractography. Through probabilistic tractography, diffusion tensor images can be used to map comprehensive neuronal network of white matter pathways of the brain. Comparisons between neural networks of mouse and human brains showed that various clinical tests on mouse models were able to simulate disease pathology of human brains, increasing the importance of clinical mouse brain studies. However, differences between brain size of human and mouse brain has made it difficult to achieve the necessary image quality for analysis and the conditions for sufficient image quality such as a long scan time makes using live samples unrealistic. In order to secure a mouse brain image with a sufficient scan time, an Ex-vivo experiment of a mouse brain was conducted for this study. Using FSL, a tool for analyzing tensor images, we proposed a semi-automated segmentation and tractography analysis pipeline of the mouse brain and applied it to various mouse models. Also, in order to determine the useful signal-to-noise ratio of the diffusion tensor image acquired for the tractography analysis, images with various excitation numbers were compared.

• Korean Journal of Food Science and Technology
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• v.50 no.2
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• pp.225-237
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• 2018

This study was conducted to compare nutritional analysis and neuroprotective effect of 5 cultivars of Diospyros kaki (Dungsi, Godongsi, Gojongsi, Gabjubaekmok, and Bansi). In nutritional analysis, three free sugars: sucrose, glucose, and fructose, and six fatty acids: tartaric acid, hexadecanoic acid, palmitic acid, oleic acid, octadecenamide, and octadecane, were detected. Potassium and phosphorus levels were the highest in inorganic component analysis, and glutamic acid and aspartic acid were the highest contents in amino acid analysis. Vitamin C was detected in all cultivars. Total phenolic content was the highest in Dungsi. Antioxidant activities such as ABTS (3-ethylbenzothiazoline-6-sulfonic acid), DPPH (1,1-diphenyl-2-picrylhydrazyl) radical scavenging activities, FRAP (ferric reducing/antioxidant power), and MDA (malondialdehyde) inhibitory effect were the highest in Gabjubaekmok. Acetylcholinesterase inhibitory activity, cell viability, intracellular reactive oxygen species (ROS) accumulation, and lactate dehydrogenase (LDH) release were measured to confirm the neuroprotective effect in MC-IXC cells. Gabjubaekmok showed significant acetylcholinesterase (AChE) inhibition and neuroprotection.

• Journal of Life Science
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• v.19 no.1
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• pp.21-33
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• 2009

In this study, we investigated the effect of Rhei Rhizoma (RR; 大黃) water extract on gene expression in a hypoxia model of cultured rat hippocampal neurons. RR water extract $(2.5{\mu}g/ml)$ was added to the culture media on day 10 in vitro (DIV10), and a hypoxic shock (2% $O_2$/5% $CO_2$, $37^{\circ}C$, 3 h) was given on DIV13. After maintaining the cultures in normoxia for 24 hr, total RNA was isolated and used for microarray analysis. The MA-plot indicated that most genes were up- or downregulated within 2-fold. There were more downregulated genes (725 ea) than upregulated ones (472 ea) when larger than Global M value 0.2 (i.e., >15% increase) or smaller than Global M value -0.2 (i.e., >15% decrease) were considered. Antiapoptosis genes such as Tegt (2.4-fold), Nfkb1 (2.4-fold) Veg (1.8-fold), Ngfr (1.6-fold) were upregulated, while pro-apoptosis genes such as Bad (-64%), Cstb (-66%) were downregulated. Genes for combating environmental stress (stress response genes) such as Defb3 (2.7-fold), Cygb (2.2-fold), Ahsg (2.18-fold), Alox5 (2-fold) were upregulated. Genes for cell proliferation (cell cycle-related genes) such as Erbb2 (1.84-fold), Mapk12 gene (1.8-fold) was upregulated. Therefore, RR water extracts upregulate many pro-survival genes while downregulating many pro-death genes. It is interpreted that these genes, in combination with other regulated genes, can promote neuronal survival in a stress such as hypoxia.

• Korean Journal of Clinical Laboratory Science
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• v.48 no.2
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• pp.144-152
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• 2016

Carotid-artery stenosis may reduce cerebral perfusion, and affect cerebral neuronal activities. We examined the question of whether the recovery of cerebral-perfusion reserve after carotid-artery stenting (CAS) can affect the EEG power-spectrum. Nineteen candidates for CAS were initially recruited. Subtraction imaging of single photon emissary computerized tomography (SPECT) and an electroencephalogram (EEG) were taken twice, before and 1 month after CAS. At each time point, the EEGs were recorded before and after injection of acetazolamide (pre-ACZ EEG and post-ACZ EEG). Finally, 7 patients were enrolled after exclusion of incomplete studies. We obtained the spectral ratio (SR) of each hemisphere. SR was defined as the divided value of the power-spectrum sum of fast activities by that of slow activities. The power-spectrum values between hemispheres were compared using the inter-hemispheric index of spectral ratio (IHISR), and we examined the correlation between the power-spectrum and the cerebral-perfusion reserve. Cerebral-perfusion reserve improved after CAS on the stent side in 6 of 7 patients. In 3 patients with unilateral carotid-artery stenosis, CAS increased SR on the pre-ACZ EEGs, and IHISR on the post-ACZ EEGs. The increases of SR and IHISR were concordant with the increment of cerebral-perfusion reserve. In contrast, the results in the other patients with bilateral stenosis showed complex patterns. The SR of pre-ACZ EEGs and IHISR of post-ACZ EEGs may be useful electrophysiological markers for the blood-flow reserve after CAS in patients with unilateral carotid-artery stenosis, but not in those with bilateral stenosis.

• Neonatal Medicine
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• v.16 no.2
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• pp.221-233
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• 2009

Purpose: Erythropoietin (EPO) has neuroprotective effects in many animal models of brain injury, including hypoxic-ischemic (HI) encephalopathy, trauma, and excitotoxicity. Current studies have demonstrated the neuroprotective effects of EPO, but limited data are available for the neonatal periods. Here in we investigated whether recombinant human EPO (rHuEPO) can protect the developing rat brain from HI injury via modulation of NMDA receptors. Methods: In an in vitro model, embryonic cortical neuronal cell cultures from Sprague-Dawley (SD) rats at 19-days gestation were established. The cultured cells were divided into five groups: normoxia (N), hypoxia (H), and 1, 10, and 100 IU/mL rHuEPO-treated (H+E1, H+ E10, and H+E100) groups. To estimate cell viability and growth, a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) assay was done. In an in vivo model, left carotid artery ligation was performed on 7-day-old SD rat pups. The animals were divided into six groups; normoxia control (NC), normoxia Sham-operated (NS), hypoxia-ischemia only (H), hypoxia-ischemia+vehicle (HV), hypoxia-ischemia+rHuEPO before a HI injury (HE-B), and hypoxia-ischemia+rHuEPO after a HI injury (HE-A). The morphologic changes following brain injuries were noted using hematoxylin and eosin (H/E) staining. Real-time PCR using primers of subunits of NMDA receptors (NR1, NR2A, NR2B, NR2C and NR2D) mRNA were performed. Results: Cell viability in the H group was decreased to less than 60% of that in the N group. In the H+E1 and H+E10 groups, cell viability was increased to >80% of the N group, but cell viability in the H+E100 group did not recover. The percentage of the left hemisphere area compared the to the right hemisphere area were 98.9% in the NC group, 99.1% in the NS group, 57.1% in the H group, 57.0% in the HV group, 87.6% in the HE-B group, and 91.6% in the HE-A group. Real-time PCR analysis of the expressions of subunits of NMDA receptors mRNAs in the in vitro and in vivo neonatal HI brain injuries generally revealed that the expression in the H group was decreased compared to the N group and the expressions in the rHuEPO-treated groups was increased compared to the H group. Conclusion: rHuEPO has neuroprotective property in perinatal HI brain injury via modulation of N-methyl-D-aspartate receptors.

• Applied Microscopy
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• v.35 no.3
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• pp.121-128
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• 2005

It has been known that ras signaling transduction leads to cell proliferation and migration including various adaptor molecules. Dynamin protein has been implicated in the formation of nascent vesicles in both the endocytic and secretory pathways. Dynamin was classified into three isoforms: dynamin I is only expressed in neuronal tissue, dynamin II is expressed ubiquitously in all tissue but that of dynamin III is confined to testis. We have reported in previous study that Grb2, binding to ras, was associated with dynamin II in NIH3T3 cells. Therefore we have tried to identify the relative expression of dynamin II according to overexpressed ras protein in ras oncogene transfected cells (NIH3T3 (ras)). For the detection of differential expression of dynamin II, we have used immunofluorescent staining and western blot methods in NIH3T3 and NIH3T3 (ras) cells. Next we have described the morphological differences between NIH3T3 and NIH3T3 (ras) cells using SEM and TEM. From these experiments dynamin II was highly expressed in NIH3T3 (ras) cells. NIH3T3 cells was transformed to more spindle shape with many cell process by transfection of ras oncogene. Moreover dynamin II was more concentrated in endocytotic membrane of the NIH3T3 (ras) cells compared to that of NIH3T3 cells. The present results suggested that dynamin II may involve the intermediate messenger in Ras signaling transduction pathway.

• Applied Microscopy
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• v.41 no.2
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• pp.109-116
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• 2011

Polycystic ovary syndrome (PCOS) is hormonal imbalance condition as the endocrine and metabolic disorder that induces the infertility and various complications in reproductive age women. Estradiol valerate (EV) is used hormone replacement therapy in menopausal women and is reported that excessive administration of EV induces the PCOS. Nerve growth factor (NGF) is the factor to regulate the survival and maturation of developing neuronal cell and is also synthesized in ovary. And NGF is overexpressed in EV-induced polycystic ovary (PCO) as previously reported. Therefore, this study examined the possibility of NGF as can be used the biological marker in diagnosis of PCOS, the hormonal imbalance condition, using PCO and CHO (chinese hamster ovarian) cell lines. The concentration of EV treatment is optimized a 1 mg as not influence on the proliferation of CHO cell but 2 mg and 3 mg of EV treatment have the inhibition effect at initial stage. The morphological change was not observed in CHO cell after dose dependent manner treatment of EV. Expression of NGF mRNA and protein is significantly increased at 30 min after EV treatment in CHO cells compared to that of control. And NGF protein expression is strongly increased in PCO tissue, which observed many follicular cysts compared to normal ovary tissue. Taken together, overexpression of NGF may be act as a molecule to induce an abnormal development of follicle, suggesting that NGF can be used as a biological marker in diagnosis of PCOS.

• Applied Microscopy
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• v.38 no.2
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• pp.141-148
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• 2008

Nitric oxide (NO) is an important regulator of renal blood flow, glomerular hemodynamics, and tubule transport processes in the kidney. There is also evidence that NO is involved in cell cycle regulation and mitotic division. During development the nNOS expression pattern differs from that observed in adult animals. However, little is known about temporal and spatial patterns of nNOS expression in the developing kidney. The purpose of this study was to establish the time of expression and the distribution of nNOS in the developing rat kidney. Kidneys from 14-, 16-, 17-, 18-, and 20-day-old fetuses, 1-, 4-, 7-, 14-, and 21-day-old pups, and adult animals were preserved and processed for immunohistochemistry. In the adult kidney, nNOS was detected in the parietal epithelium of Bowman s capsule, macula densa, descending thin limb and inner medullary collecting duct. nNOS immunoreactivity appeared first in the distal tubule anlage at 15 days of gestation, and in all epithelial cells of developing thick ascending limbs (TAL) as well as macula densa of 17- and 18-day-old fetuses. From 20 days of gestation to 14 days after birth, nNOS was expressed in the newly formed cortical TAL, which are located in the medullary ray, whereas in mature TAL of juxtamedullary nephrons, nNOS immunolabeling gradually decreased in intensity and became restricted to the macula densa. In inner medullary collecting ducts, nNOS immunoreactivity appeared first at 7 days after birth in the papillary tip and gradually ascended to the border between outer and inner medulla. In the descending thin limb and parietal epithelium of Bowman's capsule, weak nNOS immunoreactivity was observed at 14 days after birth and labeling gradually increased to adult levels at 21 days after birth. These results suggest that differential expression of nNOS in the developing kidney is an important physiological regulator of renal function during kidney maturation.