Cancer development is accompanied by genetic events like losses, gains and amplification of certain chromosome regions or alterations of chromatin structure. Array-based CGH (Array-CGH) is a highly comprehensive, sensitive and fast technique to allow investigation of general changes in target oncogenes and tumor suppressor genes. Recently, the prevalence of colon cancer is rapidly increasing in Korea and now it is the fourth leading cause of cancer death. So, the purpose of this study is to examine genomic alterations in colon cancer cell lines and to search novel genes which might be related to the development of colon cancer. In this study, genomic alterations are analyzed by using array-CGH in three colon cell lines from Korean, SNU-81, SNU-407 and SNU-1047. We observed numerous chromosomal imbalances from all cell lines. The common chromosomal gains were observed in 1p36.33, 1q22, 1q32.1, 2q35, 8p12, 8q22.3, 14q32.33, 16p13.3, and 16q24. Common chromosomal losses were found in 4q22.1, 9q13, 14q21.1, 14q32.33, 20p12.1, Xq21.1, and Yq11.223. Gains of 1p, 2q, 8p, and 8q or losses of 4q, 14q and 20p are already known to be associated with the colon cancer development. For gene alterations, we could see gains of some genes such as ELF3 and AAMP, which were already reported to be associated with colon cancer. Also, we could find some gene alterations which were known to be associated with other cancer types. These genes were GON4L, RNPEP, TMBIM1, TIMM17A, GPBAR1, PPP1R13B and SOX8. Besides, we found alterations of new genes such as PKND and LEPROTL1. The association of these genes with colon cancer is first demonstrated here. These genes may be the novel candidate genes functioning in the development of colon cancer. In conclusion, array-CGH demonstrated the complexity of genetic aberrations in several colon cell lines. These data about the patterns of genomic alterations could be a basic step for understanding more detailed genetic events in the carcinogenesis and also provide information about possible target genes for diagnosis and treatment in colon cancer.
Cell Line ; Chromatin ; Colon ; Colonic Neoplasms ; Genes, Tumor Suppressor ; Korea ; Nucleic Acid Hybridization ; Oncogenes ; Prevalence

Intranasal administration provides a method of bypassing the blood brain barrier, which separates the systemic circulating system and central interstitial fluid, and directly delivering drugs to the central nervous system. This method also circumvents first-pass elimination by the liver and gastrointestinal tract. In the present study, the authors investigated intranasal siRNA delivery efficiency by using FITC-labeled transfection control siRNA and a genespecific siRNA. The localization of fluorescence-tagged siRNA revealed that siRNA was delivered to cells in the olfactory bulb and that the level of the siRNA target gene (alpha B-crystallin) was significantly reduced in the same area. siRNA was delivered to processes as well as nuclei and cytoplasm. At 12 hrs after intranasal delivery, siRNA-mediated target gene reduction was observed in other more distally located brain regions, for example, in the amygdala, entorhinal cortex, and hypothalamus. Target gene knockdown was demonstrated by double immunohistochemistry, which demonstrated alpha B crystallin expression depletion in more than 70% of cells at 12 hrs after the intranasal delivery. siRNA-mediated target gene suppression was detected not only in neurons but in glia, for example, astrocytes. These results indicate that intranasal siRNA delivery offers an efficient means of reducing specific target genes in certain regions of the brain and of performing gene knockdown-mediated therapy.
Administration, Intranasal ; alpha-Crystallin B Chain ; Amygdala ; Astrocytes ; Blood-Brain Barrier ; Brain ; Central Nervous System ; Cytoplasm ; Entorhinal Cortex ; Extracellular Fluid ; Gastrointestinal Tract ; Gene Knockdown Techniques ; Hypothalamus ; Immunohistochemistry ; Liver ; Neuroglia ; Neurons ; Olfactory Bulb ; RNA, Small Interfering ; Transfection

Lithium-induced neuroprotection are complex and may include the neurogenesis and anti-apoptotic events. Recently, several studies have suggested the possibility that replacement of external Na+ with lithium which induced activations of sodium transporters such as the Na+/H+ exchanger 1 (NHE1) and electrogenic Na+/HCO3 -cotransporter (eNBC). Thus, alteration of sodium transporters could be associated with neurogenesis and anti-apoptotic actions of lithium even though the drug's therapeutic mechanisms remain obscure. The present study was undertaken to examine the changes of protein of NHE1 and eNBC after lithium treatment in normal and ischemic rats can regulate neurogenesis and/or apoptosis in dentate gyrus (DG). Lithium treatment was produced by pellet of standard diet containing 60 mmol/dL lithium for 25 days. The serum concentrations of lithium were found to be 0.76+/-0.2 mEq/L which is therapeutic dose of clinical practice. Immunoblotting analyses revealed that the NHE1 was significantly increased (259+/-8% of controls, n=4, P<0.01) whereas eNBC was unchanged (103+/-8%) compared with controls (n=4) after lithium treatment. Immunohistochemical studies demonstrated that bromodeoxyuridine (BrdU)-positive cells in the lithium-treated DG(n=3) were significantly higher compared with those in controls (n=3). In ischemia-reperfusion rats (n=6), terminal deoxynucleotidyl transferase dUTP nick end labelling (TUNEL) staining revealed apoptotic granule cells with ischemia-reperfusion rats while no apoptotic granule cells were showed with pretreatment of lithium. These findings suggest that significant increase of NHE1 after lithium treatment may at least partly contribute the neurogenesis and anti-apoptosis of DG via increased intracellular pH and volume increase. Therefore, lithium treatment may have therapeutic potential for ischemic stroke via neurogenesis and anti-apoptotic actions
Animals ; Apoptosis ; Bromodeoxyuridine ; Dentate Gyrus ; Diet ; DNA Nucleotidylexotransferase ; Hydrogen-Ion Concentration ; Immunoblotting ; Lithium ; Neurogenesis ; Rats ; Sodium ; Stroke

Whole body of a Korean male cadaver was serially milled to make sectioned images. Segmentation of various anatomical structures can expand the utilization of the sectioned images such as three-dimensional (3D) reconstruction of the structures of real human. Following previous outlining of lower limb's structures, we decided to make segmented images of upper limb's structures in detail. Ninety-one structures (a skin, 32 bones, 49 muscles, 6 arteries, and 3 nerves) in the left upper limb were segmented in 628 sectioned images. While doing this, we developed more efficient technique for segmentation. To draw the outlines of various structures more quickly, sectioned images were filtered first and then outlines were drawn by 'quick selection' tool and other tools on the Photoshop. Also, outlines were automatically generated by interpolation using Combustion software. We made coronal and sagittal segmented images, browsing software of the serially sectioned images, volume 3D images, and surface 3D images for verifying segmentation. These segmented and sectioned images of the upper limb are expected to help other researchers make 3D images and various software of upper limb and to have widespread applications in both medical learning and research.
Arteries ; Cadaver ; Humans ; Imaging, Three-Dimensional ; Learning ; Male ; Muscles ; Skin ; Upper Extremity

Inducible nitric oxide synthase (iNOS) has been known to be involved in the various physiological metabolim and has been attracting topic. However, there are extensive differences in the reports about the localization of iNOS expression. To resolve this discrepancy, we compared immunohistochemical data from four iNOS antibody produced by different company (Chemicon, CH; Sigma, SI; Transduction Laboratories, TL; Upstate, UP), and NADPHdiaphorase (NADPH-d) enzyme-histochemical results using light- and transmission electorn-microscope in the lipopolysaccharide (LPS)-treated rat kidney. Electron microscopical examination revealed two different distribution of the NADPH-d reaction product. In the majority of NADPH-d reaction-positive cells, reaction depositions were restricted to the mitochondia, and in the cells of macula densa, descending thin limb (DTL), capsular epithelium (CE) and interstitial wandering cells (WC), NADPH-d positivities were found in the cytoplasm. In immunohistochemical results from LPStreated animal, DTL, CE and WC were positively stained with TL and UP antibodies but with CH and SI antibodies. We conclude that NADPH-d histochemistry may be usefull for identifing the iNOS-positive cells morphologically.
Animals ; Antibodies ; Cytoplasm ; Electrons ; Epithelium ; Extremities ; Kidney ; Nitric Oxide Synthase Type II ; Rats

Chios gum mastic (CGM) is a resin produced from the stem and leaves of Pistiacia lentiscus L var chia, a plant which grows only on Chios Island in Greece. CGM has been used for many centuries as a dietary supplement and folk medicine for stomach and duodenal ulcers in many Mediterranean countries and is also known to induce cell cycle arrest and apoptosis in some cancer cells. This study was undertaken to investigate the alteration of the cell cycle and induction of apoptosis by CGM treatment on human osteosarcoma (HOS) cells. The viability and the growth inhibition of HOS cells were assessed by the MTT assay and clonogenic assay respectively. The hoechst staining, TUNEL assay and DNA electrophoresis were conducted to observe the HOS cells undergoing apoptosis. HOS cells were treated with CGM, and Western blotting, immunocytochemistry, confocal microscopy, FACScan flow cytometry, mitochondrial membrane potential change and proteasome activity were conducted. CGM treatment of HOS cells was found to result in a dose- and time-dependent decrease in cell viability, a dose-dependent inhibition of cell growth, and apoptotic cell death. Tested HOS cells also showed several lines of apoptotic manifestation and G1 arrest in cell cycle progression. In summary, this study clearly demonstrated that CGM induces G1 cell cycle arrest via the modulation of cell cycle-related proteins, and apoptosis via proteasome, mitochondrial and caspase cascades in HOS cells. Therefore, our data provide the possibility that a natural product, CGM could be considered as a novel therapeutic strategy for human osteosarcoma.
Apoptosis ; Blotting, Western ; Cell Cycle ; Cell Cycle Checkpoints ; Cell Death ; Cell Survival ; Dietary Supplements ; DNA ; Duodenal Ulcer ; Electrophoresis ; Flow Cytometry ; G1 Phase Cell Cycle Checkpoints ; Gingiva ; Greece ; Humans ; Immunohistochemistry ; In Situ Nick-End Labeling ; Medicine, Traditional ; Membrane Potential, Mitochondrial ; Microscopy, Confocal ; Osteosarcoma ; Plants ; Proteasome Endopeptidase Complex ; Proteins ; Resins, Plant ; Stomach

Neuregulin-1 (NRG1) signaling participates in numerous neurodevelopmental processes. Although ErbB4, a key NRG1 receptor, is expressed in multiple regions in the adult animal brain, little is known about its expression in aged human brain. We show that ErbB4 immunoreactivity was shown regional difference in the hippocampus of age-matched control and that the distribution of these molecules was altered in Alzheimer's disease (AD) brains. Immunohistochemical characterization of the distribution of ErbB4 receptor in the hippocampus relative to pathology staging were performed in age-matched control (Braak stage I/II, n=5), early AD (Braak stage III/IV, n=5) and advanced AD(Braak stage V/VI, n=10). The intensity of ErbB4 immunoreactivity was higher in neurons of the CA2 than that in CA1 or CA3 in the age-matched control. Particularly, in the early AD, ErbB4 immunoreactivity was significantly increased in the apoptotic cells of the CA2 field. In the advanced AD, ErbB4 immunostaining was more intense in the apoptotic cell of the CA2 field. In the dentate gyrus (DG), ErbB4-positive granular cell density was gradually increased in proportion to the progression of pathology of AD brains. We have also found that ErbB4 immunostaining was increased in the nucleus, suggesting that the presenilin-dependent cleavage of ErbB4 generates the soluble ErbB4 ICD (intracellular domain) that translocalized to the nucleus. Together, these results provide the immunohistochemical analysis of ErbB4 receptor in the human hippocampus staged by the progression of pathology of AD.
Adult ; Aged ; Alzheimer Disease ; Animals ; Apoptosis ; Brain ; Cell Count ; Dentate Gyrus ; Hippocampus ; Humans ; Neuregulin-1 ; Neurons

Ischemic preconditioning is the earlier stress adaptive response that occurs during repeated episodes of the brief ischemia and reperfusion. It is now well known that this adaptive response can render the neuron more tolerant to the subsequent potential lethal ischemic injury. Although the selective mitochondrial K+ (ATP) channel opener induces protective effects similar to that of ischemic preconditioning, the underlying mechanism is not known yet. The purpose of this study was to investigate the mechanism of neuroprotective effect of diazoxide, a mitochondrial K+ (ATP) channel opener, pretreatment on a focal cerebral ischemic injury of rat brain. Thirthy-four Sprague-Dawley rats were used. Animals were randomly divided into normal control group, middle cerebral artery (MCA) permanent occulusion group (experimental control group), and diazoxide pretreated group. Animals were sacrified at 2 hours or 24 hours after MCA occulusion injury. For inducing the focal cerebral ischemic injury, the left MCA was occuluded by modified Longa's method. Diazoxide (3 mg/kg) was administrated through the femoral artery at 15 minutes earlier to surgical procedures. TTC-stained brain sections of experimental group showed a remarkable infarct injury in the ipsilateral cerebral cortex and striatum. However, the infarction volume of the diazoxide pretreated group was significantly reduced. Accordingly, the number of neurons undergoing eosinophilic degeneration and nuclear chromatin condensation was reduced by diazoxide pretreatment. TUNEL-positive neurons were not detected at 2 hours after MCA permanent occlusion but lots of them were observed at 24 hours. The number of c-fos immunoreactive neurons was remarkably increased at 2 hours following MCA permanent occulusion and reduced to the basal level at 24 hours in both experimental control and diazoxide pretreated group. However, the number of Bcl-2 or pAkt immunoreactivitive neurons of the diazoxide pretreated group outnumbered those of the experimental control group at all timepoints in our experiment. In conclusion, the pretreatment of diazoxide, K+ channel opener, could have europrotective effects on ischemic neurons by upregulating the expression of anti-apoptotic proteins, like Bcl-2 or pAkt.
Animals ; Apoptosis ; Apoptosis Regulatory Proteins ; Brain ; Brain Ischemia ; Cerebral Cortex ; Chromatin ; Diazoxide ; Eosinophils ; Femoral Artery ; Infarction ; Ischemia ; Ischemic Preconditioning ; Middle Cerebral Artery ; Models, Animal ; Neurons ; Neuroprotective Agents ; Rats ; Rats, Sprague-Dawley ; Reperfusion

The organ weight is one important indicator to discern normal condition from abnormal in forensic pathology as well as in clinical medicine. This study includes organ weights of Korean population, which can be fundamental sources to be analyzed comparatively with other ethnic groups. Seven organs (heart, lung, liver, kidney, spleen, thyroid gland, and brain), which were harvested from 526 Korean adults (369 males and 157 females) during ordinary postmortem examination, were weighed. All of the organs in males were heavier than those in females. With variables controlled, the organ weights of hearts, spleens, and thyroid glands in males were not different from those in females, and the rest of organs were heavier in males than females. Mean weight of left kidneys was higher than that of right ones (P<0.05). All of the organs but heart became lighter in weight, as one got older. The liver and kidney weights of the middle-aged adults were heavier than those of any other age groups (P<0.05). The weights of all organs except for lungs and cerebrums were more related to body weight than height. These results are considered for useful anatomical data to understand the disease properties in Koreans.
Adult ; Autopsy ; Body Weight ; Cerebrum ; Clinical Medicine ; Ethnic Groups ; Female ; Forensic Pathology ; Heart ; Humans ; Kidney ; Liver ; Lung ; Male ; Organ Size ; Spleen ; Thyroid Gland ; Weights and Measures

Transmission electron microscopy (TEM) provides high resolution images, which are useful in studying ultrastructure of cells and tissues. We have to use very thin section about 60~100 nm thickness due to poor penetration power of the conventional TEM at 100 kV. To overcome this limitation, TEMs using higher accelerating voltage have been developed. TEMs can be categorized into conventional TEM, intermediate TEM, high voltage TEM (HVEM), and ultrahigh voltage TEM according to their accelerating voltage. HVEM using 500~1,000 kV has an enough penetration power to observe thick specimen up to 3~4 micro, which is useful understanding 3 dimensional configuration of the cell and tissue. HVEM was built up in Korea Basic Science Institute (KBSI, Daejeon, Korea) at 2004, maximum accelerating voltage is 1.3 MV in Korea. Many results showed up to the present various fields of science such as medical science, biology, agriculture and so on. Here, we briefly summarize recent biomedical applications of HVEM to provide an insight of HVEM for morphologist.
Agriculture ; Biology ; Electrons ; Korea ; Microscopy, Electron ; Microscopy, Electron, Transmission