Bile acids and their synthetic derivatives induced apoptosis in various kinds of cancer cells and anticancer effects. It has been reported that the synthetic chenodeoxycholic acid (CDCA) derivatives showed apoptosis-inducing activity on various cancer cells in vitro. It wasn't discovered those materials have apoptosis-inducing effects on G361 human melanoma cells. The present study was done to examine the synthetic bile acid derivatives, HS-1199 and HS-1200, induced apoptosis on G361 cells and such these apoptosis events. The viability of G361 cells was assessed by the MTT assay. Induction of apoptosis was confirmed by DNA electrophoresis and Hoechst staining. Westen blot analysis and immunofluorescent staining were performed to study the alterations in expression level and translocation of apoptosis-related proteins. Proteasome activity and mitochondrial membrane potential (MMP) change were also assayed. Tested G361 cells showed several lines of apoptotic manifestation such as activation of caspase-3, DFF and PARP, DNA degradation (HS-1200 only), nuclear condensation, inhibition of proteasome activity, reduction of mitochondrial membrane potential, and the release of cytochrome c and AIF to cytosol. Between two synthetic derivatives, HS-1200 showed stronger apoptosis-inducing effect than HS-1199 did. Taken collectively, we here demonstrated for the first time that synthetic CDCA dedrivatives induce apoptosis of human melanoma cells through the proteasome, mitochondria and caspase pathway. Therefore our data provide the possibility that HS-1200 could be considered as a novel therapeutic strategy for human melanoma cells from its powerful apoptosis-inducing activity.
Apoptosis* ; Bile ; Bile Acids and Salts ; Caspase 3 ; Chenodeoxycholic Acid* ; Cytochromes c ; Cytosol ; DNA ; Electrophoresis ; Humans* ; Melanoma* ; Membrane Potential, Mitochondrial ; Mitochondria ; Proteasome Endopeptidase Complex

In the present study, we investigated influences of glycogen synthase kinase (GSK) 3beta on the development and/or progression of amyotrophic lateral sclerosis (ALS). We used transgenic mice expressing a human Cu/Zn superoxide dismutase mutant (SOD1G93A) as an in vivo model of ALS and examined expressional changes of GSK3beta immunohistochemically in the spinal cord, brain stem and cerebellum. With these experiments we demonstrate that the neurons in these regions of symptomatic SOD1G93A transgenic mice showed increased GSK3beta immunoreactivities compared with wild-type SOD1 transgenic mice. In contrast to symptomatic SOD1G93A transgenic mice, few GSK3beta immunoreactivity changes were detected in 8w- and 13w-old presymptomatic SOD1G93A transgenic mice. These data suggest the possibility that GSK3 functions as a modulating factor of apoptosis-related alterations in ALS and that GSK3beta exert differential functions in the development and/or progression of ALS. But the exact functional significances of these changes require further elucidation.
Amyotrophic Lateral Sclerosis ; Animals ; Brain Stem ; Central Nervous System* ; Cerebellum ; Glycogen Synthase Kinases* ; Glycogen Synthase* ; Glycogen* ; Humans ; Mice ; Mice, Transgenic* ; Neurons ; Spinal Cord ; Superoxide Dismutase

The effect of peripheral nerve on limb growth and maturation has received little attention after the limb differentiation stage. This study investigated the effects of paralysis (loss of function) on growth of bones in the hind limb of Hymenochirus boettgeri. Complete denervation of the right sciatic nerve was performed at stages 63 and 66, animals were sacrificed after 6 and 7 (Groups I and II) and 12 and 13 (Groups III and IV) weeks. Denervation was assessed by degree of paralysis. Specimens were cleared and double stained with alcian blue and alizarin red for cartilage and bone. Length and area of right and left femurs and length of right and left tibia were measured by using an image analyzer program after photographing, and the ratio of right to left femur length and area was calculated. There were no gross morphological differences between the control and sham groups. In the experimental groups, the ratio of femur length was 93.71% and 95.70% in Group I and II, and 96.12% and 96.06% in Group III and IV. The ratio of tibia length was 94.05% and 96.15% in Group I and II, and 98.12% and 98.22% in Group III and IV. The ratio of femur area was 90.43% and 95.61% in Group I and II, and 96.08% and 95.73% in Group III and IV. Comparison between control and experimental groups showed statistically significant (p<0.05). There was a histologically maturational delay in proximal end of denervated hind limb, comparing with opposite side. These results suggested that denervation of sciatic nerve affect directly the growth and maturation of hind limb bones in Hymenochirus boettgeri and loss of critical period of bony maturation after denervation.
Alcian Blue ; Animals ; Bone Development ; Cartilage ; Critical Period (Psychology) ; Denervation* ; Extremities* ; Femur ; Paralysis ; Peripheral Nerves ; Sciatic Nerve ; Tibia

Retinoids play an important role in growth, reproduction and differentiation. Recently, retinoids have been used to both protect and treat from various animal models of carcinogenesis. In this study the effect of N-(4-hydroxyphenyl) retinamide (fenretinide) on viability of human neuroblastoma cell lines were evaluated. For the evaluation of apoptosis of human neuroblastoma cell lines by fenretinide. MTT assay, cytoplasmic DNA fragmentation, TUNEL stain, and Western blot analysis were performed. In MTT assay, fenretinide inhibited the proliferation of CHP134, IMR32 and SH-SY5Y but not in PC12 cells. Cytoplasmic DNA fragmentation was induced by treament of fenretinide (10 micrometer) for 48 h in IMR32 cells. PARP cleavage was detected by Western blot analysis after 16 h of treatment of fenretinide in CHP134, IMR32 and SH-SY5Y. These fenretinide effects on growth inhibition and increased apoptosis followed to the time dependent manner. The fenretinide treatment did not affect the phosphorylation of MAP kinases (ERK, JNK, p38). There was no change of Bcl-x and Bad expression after treatment of fenretinide (1 micrometer) in neroblastoma cell lines. Pretreatement of PD98059, SB203580, LY294002, or genistein also did not affect fenretinide-induced PARP cleavage in neuroblastoma cell lines. From these results, the fenretinide-induced apoptosis is due to the PARP cleavage which occured MAP kinase signal cascades independently.
Animals ; Apoptosis* ; Blotting, Western ; Carcinogenesis ; Cell Line* ; Cytoplasm ; DNA Fragmentation ; Fenretinide* ; Genistein ; Humans* ; In Situ Nick-End Labeling ; Models, Animal ; Neuroblastoma* ; PC12 Cells ; Phosphorylation ; Phosphotransferases ; Reproduction ; Retinoids

Interleukin-6 (IL-6) and its receptor are presumed to play important roles in the developing nervous system. However, little is known about their potential role(s) in the developing kidney. To investigate this, we have studied the expression of IL-6 and its receptor (IL-6R) in the developing rat kidney. Kidneys from 16- (F16), 18- and 20-day-old (F20) fetuses, 1- (P1), 3- (P3), 7- (P7) and 14-day-old (P14) pups, and adult rats were extracted. Renal expressions of IL-6 and its receptors were examined by immunohistochemistry and in situ hybridization respectively. Il-6 protein already appeared in F16. The early stage of renal development before birth, IL-6 showed strong immunoreactivity in the ureteric bud, metanephric mesenchymal cells (MMC) and developing glomerulus. The expression pattern of IL-6 in nephrogenic zone are very similar even after birth. In matured nephron after birth, IL-6 immunoreactivities were detected in distal tubules strongly, and collecting ducts moderately and thick ascending limb weekly. IL-6R hybridization signals have also already appeared in 16-day old fetal kidney. Before birth, IL-6R mRNAs were expressed in ureteric bud, MMC and developing glomerulus. In the matured nephron after birth, IL-6R mRNA was expressed in the thick ascending limb, distal tubules, collecting ducts and S3 segment of proximal tubule. These results suggest that IL-6 and its receptor may be involved in regulation of nephron formation in nephrogenic zone of rat, and play a role in distal nephron including collecting duct after birth.
Adult ; Animals ; Extremities ; Fetus ; Humans ; Immunohistochemistry ; In Situ Hybridization ; Interleukin-6* ; Kidney* ; Nephrons ; Nervous System ; Parturition ; Rats* ; RNA, Messenger ; Ureter

Homeostasis of the body is maintained by balance of cell renewal, differentiation, and death. Dexamethasone has been used long time in field of MSCs research as differentiation inducer while, in contrast, there has been little work on the effect of dexamethasone on apoptosis and proliferation of MSCs. To determine the effect of dexamethasone on apoptosis and proliferation, MSCs were cultivated with (Dex) or without (Con) dexamethasone for 3 weeks. During this period weekly cell count, DNA assay, mitosis, apoptosis as well as alkaline phosphatase assay observation were recorded. DNA and cell number of Dex group was lower than Con group in early period but exceed at 3th week. There is no significant difference in mitosis between both groups whereas apoptosis frequency in Dex group was lower than that of in Con group. These results indicate that dexamethasone influences cell proliferation of MSCs in long term confluent culture by suppression of apoptosis.
Alkaline Phosphatase ; Apoptosis ; Cell Count ; Cell Proliferation ; Dexamethasone* ; DNA ; Homeostasis ; Mesenchymal Stromal Cells* ; Mitosis

Previous studies showed that whole body vibration could prevent bone resorption and stimulate new bone formation. The aim of this study is to detect and track effect of whole body vibration for osteoporotic bone of OVX rats. 12 Female rats were used and allocated into 5 group, CON, SHAM, WBV 17, WBV 30 and WBV 45. Rats except SHAM group were ovariectomised to induce osteoporosis. Rats in WBV groups were stimulated whole body vibration at magnitude of 1 mm(peak-peak) and frequency 17 Hz, 30 Hz and 45 Hz, repectively, for 8 weeks (20 min, 5 days/week). The 4th lumbar in rats were scanned at a resolution of 35 micrometer at baseline, before stimulation, and 8 weeks by In-vivo Micro-CT. To detect and track changes of morphological characteristics in lumbar trabecuar bone of rats, structural parameters were measured and calculated from acquiring images. In the results, changes of structural characteristics of WBV group were smaller than those of CON group. Loss of quantity of trabecular bone in WBV 45 was the least. In contrast, that in WBV 17 was the biggest. These results showed that reasonable whole body vibration beneficially affected osteoporotic bones. In addition to, whole body vibration was likely to be substituted partly for drug treatment.
Animals ; Bone Resorption ; Female ; Humans ; Osteogenesis ; Osteoporosis ; Rats* ; Vibration*

This study aims to investigate the anatomical information of the sphenopalatine foramen with special reference to the bleeding control of the sphenopalatine artery and to the blockage of the pterygopalatine ganglion during functional endoscopic sinus surgery. Forty-three midsagittal sectioned Korean heads were used in the study. The mucosa on the lateral nasal wall was removed for showing the sphenopalatine foramen. The shapes of the sphenopalatine foramen were classified by 4 types. Angle from the akanthion and distances from akanthion, conchae, sphenoidal sinus and greater palatine foramen were measured. The sphenopalatine foramen was frequently found as oval shape and bilateral symmetry of 28.6%. The vertical length was 5.5 mm, the horizontal length was 5.3 mm. The most anterior point of the sphenopalatine foramen was located on 54.4 mm few and angles were between 15.9degrees and 22.1degrees from akanthion. The distances were 36.2 mm from anterior end of middle nasal concha, 6.2 mm from posterior end of middle nasal concha, 27.2 mm from greater palatine foramen. The uppermost point of the sphenopalatine foramen was located on 22.3 mm from the most superoanterior end of the sphenoidal sinus, 18.6 mm from the lowermost end of the sphenoidal sinus. This study could be useful to provide the confidence about the location of the sphenopalatine foramen during functional endoscopic sinus surgery.
Arteries ; Ganglion Cysts ; Head ; Hemorrhage ; Mucous Membrane ; Turbinates

Recently, with the increasing interests in the endoscopic nasal surgery, it is essential for practitioners to have detailed knowledges of the anatomical structures for the successful surgery. This study was undertaken to provide morphological information of the nasal turbinates as anatomical reference during endoscopic nasal surgery. A total of 43 adult cadaver heads were bisected midsagitally and evaluated morphometrically. The results were as follows: The highest nasal turbinatee were observed in 66.7% of the case. After removal of the mucosa, the distances from the akanthion to the anterior end of superior, middle, inferior nasal turbinate were 46.75 mm, 38.23 mm, 18.51 mm respectively. The distances from the akanthion to the posterior end of superior, middle, inferior nasal turbinate were 55.11 mm, 54.72 mm, 50.26 mm respectively. The lengths of the superior, middle, inferior nasal turbinate were 22.20 mm, 40.14 mm, 40.97 mm respectively. The detailed measurements of the nasal turbinates performed in this study may provide useful anatomical landmarks for the endonasal endoscopic surgery.
Adult ; Cadaver ; Head ; Humans ; Mucous Membrane ; Nasal Surgical Procedures ; Turbinates*

Ancient DNA analyses are widely used for evolutionary and phylogenetic study of mankind in anthropology and archeology. However, the DNA extraction from particularly poorly preserved ancient human samples is often unsuccessful in these analyses. In the present study, to improve the success rate of ancient DNA analysis, we introduced a high grade ancient DNA purification method using ion-exchange columns. We compared the success rate of ancient DNA analysis of this new method with that of the two methods that have been used for ancient DNA extraction, GENECLEAN(R) kit (Qbiogene) and Qiaquick column (Qiagen). Twelve ancient bone samples from Korea and Mongolia that are about 500 to 5,000 years old by an archeological estimation were used. As the DNA analysis methods, polymerase chain reaction (PCR) methods for the amplification of a mitochondrial DNA HV1 segment, a male sex determination marker DNA and M175 marker DNA that is used for the determination of O haplogroup of Y chromosome that is reportedly a common one in modern Korean people. The method developed in this study remarkably increased the success rate of DNA analysis compared with the other two methods. Using the GENECLEAN(R) kit, only two samples were amplifiable for the mitochondrial DNA, no samples for the male sex determination marker and M175 marker DNAs. Using the Qiaquick columns, nine samples were amplifiable for mitochondirial DNA, nine samples for male sex determination marker and six samples for M175 marker. The developed method allowed for the amplification of mitochondrial DNA from all samples, male sex determination marker from eight samples and M175 marker from eight samples. The results demonstrate that ion-exchange columns can be useful for the improved ancient DNA extraction in anthropology and archeology.
Anthropology ; Archaeology ; DNA* ; DNA, Mitochondrial ; Humans ; Korea ; Male ; Mongolia ; Polymerase Chain Reaction* ; Y Chromosome