L-PDMP, Ganglioside Synthesis Enhancer Protects Dopaminergic Neurons from Death by Proteasomal Inhibition and the Accumulation of alpha-synuclein / 체질인류학회지

Gangliosides are components of the membranous constituents and abundant in the nervous system. And they are implicated in a wide range of biological activities including the regulation of cell proliferation, differentiation and lysosomal activity. But they have a diverse action to induce neuronal cell death by the interaction with some ligands. The interference of their biosynthesis is accompanied by the intracellular accumulation of unwanted and neurotoxic proteins and might underlie the neurodegeneration diseases including Parkinson's disease. However the mechanism has not been elucidated. In this study, we report that the enhancement of biosynthesis of ganglioside GD3 protects the intracellular accumulation of alpha-synuclein and neuronal death. PC12 cells, dopaminergic neurons are cultured with synthetic proteasomal inhibitor (PSI, Z-lle-Glu(OtBu)-Ala-Leu-al) and L-PDMP (GD3 synthetase enhancer, L-threo-1-phenyl-2-decanoylamino-3-morpholino-1-propanol). We found that the neuronal viability was recovered by L-PDMP from the proteasomal inhibition and also the expression of activated caspase-3 and PARP was reduced. L-PDMP decreased in the intracellular accumulation of alpha-synuclein. Interestingly, PSI induced the expression of ganglioside3 in PC12 cell. Our findings suggest that proteasomal inhibition may modulate the biosynthesis of GD3 and L-PDMP protects dopaminergic neurons from death by proteasomal inhibition and the accumulation of alpha-synuclein.

Effects of Hypoxia on the Ubiquitin-proteasome System in Primary Cortical Neuronal Cell Cultures / 체질인류학회지

The ubiquitin-proteasome system is crucial in maintaining cellular growth and metabolism. Dysfunction of this system may contribute to neurodegenerative diseases, such as Parkinson's disease. But its effects on primary neurons are largely unknown. In the present study, we investigated the effects of proteasome inhibitor and hypoxia on primary neuronal cultures to determine whether proteasomal malfunction induces neuronal death. Neuronal apoptosis increased in primary cultured cortical neurons with treatment of proteasomal inhibitor in normoxic condition and in the presence or absence of proteasomal inhibitor in hypoxic condition. Also expression of PARP and activated caspase 3 increased. NF-kappaB, a key transcription factor in this system expression increased in hypoxic condition and proteasomal inhibition. Interestingly, hypoxic condition induced an expression and accumulation of alpha-synuclein in neuron, one of components of Lewy body in Parkinson's disease. Our findings determine that hypoxic condition may affect the ubiquitin-proteasome system. Furthermore, it suggests that hypoxic condition and proteasomal inhibitors are involved, at least in parts, in neurodegeneration of mouse model for Parkinson's disease.

Age-related Changes of MHC Class II Positive Dendritic Cells in Cardiac Muscle of Rat / 체질인류학회지

Cardiac dendritic cells are considered to play an important role in the immunoresponse of the heart. However, It is unknown that changes of shapes and numbers of these cells in the heart. The aim of this study is to reveal age-related changes of MHC class II positive dendritic cells in cardiac muscle of rat. Male Sprague-Dawley rats (1 month, 12 months, and 24 months old) were used in this study. Animals were deeply anesthetized with 3.5% chloral hydrate (1 mL/100 g) and hearts removed. Immunostaining was done according to standard methods used routinely. In brief, tissue sections were incubated with primary antibodies generated in mouse anti-rat MHC class II antibody for single immunostains. Tissue sections were observed by using light microscope and dendritic cells were counted. Average numbers of MHC class II-positive dendritic cells were 1.4 cells per unit area (0.2 mm2) at 1 month old rat, 2.8 cells at 12 months old rat, and 4.6 cells at 24 months old rat, and then numbers of dendritic cells were increased according to ages. According as age increases, cytoplasmic processes of MHC class II-immunoreactive dendritic cells became longer and more complex and aggregated together. It's suggested that age-related changes of MHC class II positive dendritic cells in the cardiac muscle would be related to immunity.

Effects of Recombinant EPO on Death of Cortical Neuron in Chronic Hypoxia / 체질인류학회지

Chronic hypoxia has been associated with change in neurovascular behavior, mediated, in part, by erythropoietin (EPO). EPO, a hematopoietic growth factor, could act as a neurotrophic factor. In the present study, we investigated the characteristics of EPO and erythropoietin receptor (EPOR) expressions by cortical neuron in vivo and in vitro and tested the hypothesis that EPO serves protective functions under chronic hypoxia. E18, P5 and P7 mice for 3 days and primary cultured neurons for 6 days were incubated in hypoxic conditions consisted of a mixture of 10% O2, 5% CO2, 85% N2. To study expressions of EPO, EPOR, caspases, pAKT, pERK, and PARP, immunohistochemical stainning and western blotting were carried out. In addition to expressing EPO and EPOR under normoxic conditions, neurons increased their expression of EPO and EPOR under hypoxia. The effects of recombinant EPO appeared to be mediated via the phosphatidylinositol (PI) 3- kinase-AKT pathway, correlated directly with activation of caspase 3. Also recombinant EPO decreased expression of caspase 8, but not caspase 9. Finally, recombinant EPO decreased apoptosis of cultured neurons as evaluated by expression of PARP. These data support a role for EPO in maintenance of cortical neuron under chronic hypoxia.

Inflammatory Response of Secretoneurin -Immunoreactive Nerves in the Rat Uterus after Formalin Exposure / 체질인류학회지

It is known that there are numerous chemotatic secretoneurin -immunoreactive nerve fibers and movable MHC class II -immunoreactive dendritic cells in the normal uterine cervix. And the relationships between them are not fully understood. The aim of this study is to reveal that secretoneurin could give to chemotatic influence to dendritic cells in inflammational state. Virgin female Sprague -Dawley rats (n = 20; approximately 2 months old; 200 ~250 g body weight) were used in this study. Animals (n = 10) were injected with 5% formalin (0.5 ml/day, 5 days) in experiment group. Animals were deeply anesthetized with 3.5% chloral hydrate (100 mg/kg, i.p.) and uterine cervix were removed. Immunostaining was done according to standard methods used routinely. In brief, tissue sections were incubated with primary antibodies generated in mouse anti -rat MHC class II antibody and mouse or rabbit anti -rat secretoneurin antibody for single and double immunostains. FITC for secretoneurin and rhodamine for MHC class II were used as secondary antibodies in double stains. Tissue sections were observed by using light and confocal laser scanning microscopes. The results were as follows; 1. Numerous secretoneurin -immunoreactive nerve fibers were located in the lamina propria and those were not found in the epithelium of normal rat uterine cervix. 2. MHC class II -immunoreactive dendritic cells were mainly located in the epithelium and the lamina propria of normal rat uterine cervix. 3. On the inflammation state, MHC class II -immunoreactive dendritic cells were mainly located in the lamina propria and those were not found in the epithelium of rat uterine cervix. According to above results, it is suggested that secretoneurin can give to chemotatic influence to dendritic cells in inflammational state. Therefore, secretoneurin is considered to be used for dendritic cell immunotheraphy.

Effects of Retinoic Acid on the Cell Proliferating Activity and the Expression of Fibroblast Growth Factor 2, Fibroblast Growth Factor Receptor 2 during Palatal Development of Mice / 대한해부학회지

Retinoic acid plays an important role in embryogenesis, by regulating morphogenesis, cell proliferation, differentiation, and extracellular matrix production. Also retinoic acid is a potent teratogen and induces a variety of limb and craniofacial malformations including cleft palate, that is the most common congenital malformation. Fibroblast growth factor 2 (FGF2) and fibroblast growth factor receptor 2 (FGFR2) are an important role in the secondary induction for the epithelial-mesenchymal transformation during development. Mutations in them, produce a congenital malformation in the skeletal system and the craniofacial tissue. It was of interest to explore the hypothesis of an inhibitory effect exerted by retinoic acid on the cell proliferating activity and the expression of FGF2 and FGFR2 in the developing palate in vivo. In the present study, author observed the expression of PCNA as a marker for the cell proliferating activity, FGF2 and FGFR2 to compare with developmental stages and locations in normal and retinoic acid-induced cleft palate. Retinoic acid was administered orally at gestational day (GD) 10 to ICR mice. The pregnant mice were sacrificed on GD 12, 13, 14, 15 to obtain the fetuses. Scanning electron microscope and immunohistochemistry was performed. In the retinoic acid-treated fetuses, palatal shelves did not elevate and cleft palate was induced. On GD 12, 13 in the palatal mesenchyme of the retinoic acid treated-fetuses, expression of the PCNA decreased. On GD 12 in the palatal epithelium of the retinoic acid-treated fetuses, expression of FGFR2 decreased, but after GD 13, the patterns of expression of FGFR2 were not affected. On GD 12, 13 in the palatal epithelium and mesenchyme of the retinoic acid-treated fetuses, expression of FGF2 decreased dramatically, but after GD 14, it was similar to that in the normal fetal palate. These results suggest that retinoic acid inhibits the cell proliferating activity and the expression of FGF2, FGFR2 in the palatal mesenchyme on GD 12, 13, which is critical in the developing palate, and elevation of palatal shelves is delayed and impaired. Moreover, it seems that retinoic acid inhibits the epithelial-mesenchymal transformation of epithelium. Finally, cleft palate is induced.

Neurobehavioral Tests in Workers Exposed to Ethylene Oxide / 대한산업의학회지

Ethylene oxide is widely used to sterilize heat-sensitive materials in hospital. Previous reports for neurotoxic effects of ethylene oxide have been described in animals and humans. To assess the exposure level and neurobehavioral effect of ethylene oxide, a cross-sectional study was performed to 27 nurses from central supply unit at hospital, exposed to ethylene oxide and 32 nurses as reference. Ethylene oxide was collected with using a personal air sampler and analyzed by gas chromatography for the determination of exposure level, and five items among Neurobehavioral Core Test Battery (NCTB) of World Health Organization, including Digit Span, Benton visual retention. Santa Ana dexterity, Digit Symbol and Simple reaction time, were administered to the exposed and reference group. The mean exposure level was 0.63 ppm in the exposed group and six subjects were exposed above the level of 1 ppm, which is currently regulated by the Korean Ministry of Labor. The results of neurobehavioral test in the exposed group showed significantly poorer performances in Digit Span forward and backward, Benton visual retention and Simple reaction time, comparing with the reference group. When the exposure level was divided into below and above the level of 1 ppm, there were significant differences in performance on Benton visual retentions, Digit Symbol and Simple reaction time. Also, Digit Span forward and backward showed significantly poorer performances below the level of 1 ppm, compared with reference group. Simple reaction time was still significantly delayed by the exposure level after controlling the confounding factors with multiple regression analysis. The results suggest that the periodic measurement of ethylene oxide in hospital and health care program is needed.

Studies on the Production of Nitric Oxide, Interleukin-1 and Interleukin-6 in Mice Exposed with Sarcoma 180 Cells / 대한해부학회지

This study was conducted to elucidate the biological role of the overproduced nitric oxide (NO), interleukin-l (IL-l), and interleukin-6 (IL-6) which are known to elicit inflammation, rheumatic arthritis, fever, septic shock or other fatal reactions. I investigated whether the Scarcoma 180 cells elicit NO, IL-l and IL-6 production in vivo and in vitro by measuring the NO, IL-1 and IL-6 level in the splenocyte adherent cell (AD), non-adherent cell (NAD) and whole cell (W) exposed to sarcoma 180 cells. I also measured the NO, IL-1 and IL-6 level in the plasma and peritoneal fluid of the sarcoma 180 cell-transplanted mice after 2 hours, 4 hours, 6 hours and 24 hours of incubation. 1. In the splenocyte exposed to sarcoma 180 cells, the NO production of AD and NAD increased after 2, 4, and 6 hours but decreased after 24 hours of incubation. In the whole cell, the NO production was variable; it showed increased level of synthesis at 2 hours, decreased level at 4 hours, increased level again at 6 hours, and decreased level at 24 hours of incubation. In the plasma of the sarcoma 180 cell-transplanted mice, the NO synthesis significantly increased from the 6 hours of incubation. In the peritoneal fluid, the NO production significantly increased until the 4 hours of incubation then decreased gradually. However, it showed higher level of NO production compared to the control group. 2. For the splenocyte AD cell exposed to saracoma 180 cells, the IL-1 level decreased after 6 hours of incubation. For the W cells, the IL-1 level decreased at 4 hours then increased until 24 hours of incubation. The NAD cell showed increased level of IL-I production from 2 to 24 hours. All these cells showed significantly increased level of IL-1 production compared to the control. In the plasma of the sarcoma 180 cell-transplanted mice, the IL-1 production increased more than twice the level of control from the beginning. In the peritoneal fluid, no IL-1 production was detected as in the control. 3. The IL-6 synthesis of the sarcoma 180 cell-exposed splenocyte singnificantly increased compared to the control: the AD cell showed increased level of IL-6 production after 4 hours of incubation. For the NAD cell, increased level of IL-6 was detected at 2 hours after the incubation. In the plasma of the sarcoma 180 cells transplanted mice, the IL-6 level at 2 hours after incubation was 64.22+/- 5.85 pg/ml. After 4 hours of incubation, the level decreased to 43.55+/-1.56 pg/ml. At six and 24 hours after the incubation, no IL-6 was detected. In the control, IL-6 production was not detected. In the peritoneal fluid, the IL-6 production level was 712.41+/-4.27 pg/ml after 2 hours and 225.71+/-9.74 pg/ml after 4 hours of incubation, producing singnificantly higher level of IL-6 compared to the control. After 6 hours of incubation, IL-6 level was 8.27+/-0.78 pg/ml. After 24 hours of incubation, it decreased to 1.38+/-0.39 pg/ml as time proceeds. After 24 hours of incubation, the IL-6 level was the same as the control. These results suggest that the mice which were exposed to sarcoma 180 cells, nitric oxide, interleukin-1 and interleukin-6 which may lead to inflammation, fever, sepsis and septic shock. This study also helps us better understanding of the role of cytokines in inflammatory reaction.

Differential Expression of Transforming Growth Factor-α and Epidermal Growth Factor in Developing Mouse Embryo / 체질인류학회지

The distribution of transforming growth factor-α (TGF-α) and epidermal growth factor (EGF) in developing mouse embryos of gestational age 8 to 15 days was immunohistochemically (ABC method) studied to investigate the differential expression of these growth factors. Paraffin embedded sections were immunostained with antibodies for TGF-α and EGF. Staining of TGF-α was observed in several organs derived from endoderm, mesoderm and ectoderm in 9-day-old mouse embryos, such as in the heart, optic pit, head mesenchyme, neural tube and primitive gut, and the staining became more intense in 10 to 15-day-old mouse embryos. The staining of EGF was seen in the heart and primitive gut derived from mesoderm and ectoderm respectively, in 9-day-old mouse embryos, but it was observed in other organs as well in 10 to 15-day-old embryos although the intensity was weaker. In the development of heart, immunoreactivity for TGF-α was more intense than EGF, which suggests more active involvement of TGF-α. In the lung, TGF-α staining was observed both in the bronchus and lung bud, whereas EGF staining was seen only the bronchus. In the nervous system, TGF-α was expressed more extensively and more intensively than EGF. In the developing skeletal system, TGF-α staining was stronger and the expression was observed at earlier stage compared with EGF. These results indicate that the activity of TGF-α is more potent than EGF in the development of mouse embryo in general, especially, in the development of mouse heart, nervous system, mesenchyme and skeletal system.

Morphometric Study of the Lumbar Spinal Canal in Korean Adults / 체질인류학회지

In the antero-posterior and lateral viewe of the lumbar X-ray film of 258 normal Korean adults(120 males, 138 females), the following measurement was made ; the measurement of the interpeduncular distance, anteroposterior diaameter, width of the lumbar vertebral body and height, and thickness of the pedicle. The mean value of interpeduncular distances and anteroposterior diameter of each lumbar vertebral level was measured in relation to the variables of sex and age. The value of interpeduncular distance at each level was related to the pedicle index and width of the vertebral bodies. Statistical analysis was made. The percentage of type A was 74.42%, type B 21.32%, type C 4.26%, but type D was not observed. The mean value of the interpeduncular distances of each level of the lumbar vertebra was measured. In male, Ll was 23.57mm, L2 24.29mm, L3 25.36mm, L4 27.09mm, and L5 was 29.77mm. In female, Ll was 22.24mm, L2 23.03mm, L3 24.11mm, L4 25.70mm, and L5 28.29mm. The difference between male and female was significant (P3.58). There was no significant relationship between the interpeduncular distance, anteroposterior diameter, pedicle indices and the width of the body at all levels.

Anatomical Study on the Location of the Mental Foramen in Adult Korean Mandibles / 체질인류학회지

We examined the anatomical position of the mental foramina in mandibles foramen normal adult Koreans. 1. The percentages obtained from the study of the relationships between the mental foramen and the lower teeth showed that the most common location was type lv in which the mental foramen lay at the apex of the second promolar. The foramen between thr apices of ice two premolars (type lll) and the foramen between the second premolar and the first molar (type v) occured often and less often rspectively and find no foramen mesial to the first premolar or at the apex of the first premolar and posterior of the first molar (type l, ll, vl). 2. The study of relationship of the mental foramen to the bo of the mandible revealed that mental foramen was situated closer to the lowed border of the mandibular body. The distance ratio between the mental foramen and the alveolar crest to that between the mental foramen and the lower border was approximately 1.2 : 1. The height of the mandibular body was 31.09±2.80mm on the left side and 30.97±2.48mm on the right. 3. The distance from the mandibular symphysis to the anterior border of the mental foramen measured 29.52±2.01mm on the left, 30.82±2.04mm on the right side, and from the mandibular symphysis to the posterior border of the mandibular ramus was 104.20±4.74mm on the left, 105.44±4.49mm on the right side. It indicates that the mental foramen lies approximately at one-fourth of the distance from the mandibular symphysis to 2017-04-19 the posterior border of the ramus. 4. The distance from the superior border of the mental foramen to the bottom of the lower second premolar socket was found to be positive. It was 5.46±3.09mm on the left, 5.73±3.03mm on the right side. This indicates that the bottom of the lower second premolar socket is slightly higher than the superior border of the mental foramen.