This study presents distribution of carbonic anhydrase (CA) isozymes IV and IX, membrane associated forms, and CA I and II, cytoplasmic forms, in rat parotid and submandibular glands using Western blot analysis and immunohistochemical staining. Western blot analysis demonstrated that CAs I, II and IX were found to be abundantly expressed, but CA IV was weakly expressed in parotid gland. Submandibular gland expressed abundant CAs I and II, weak CA IX, and undetectable level of CA IV. In hematoxylin-eosin staining, parotid gland was entirely composed of serous acini and their ducts while submandibular gland was mixed population of serous and mucous lobules. Most of lobules (submandibular gland proper type) contained mostly serous acini and their ducts with granular convoluted duct. Some lobules (sublingual gland type) contained mostly mucous acini with serous demilune and their ducts without granular convoluted duct. In parotid gland, CAs IV and IX were immunolocalized in duct cells and not in serous acinar cells. Immunoreactivity for CAs I and II was also detectable in duct cells. Serous acinar cells were positive for CA II, and negative for CA I. In submandibular gland, CAs IV and IX were immunolocalized in duct cells but not in acinar cells of both types of lobules. Immunoreactivity for CAs I and II was also detectable in duct cells of both types of lobules. Cells of serous acini and serous demilune were positive for CA II, and negative for CA I. Mucous cells were negative for both CAs I and II. These results demonstrate the distribution of CA isoenzymes in parotid and submandibular glands of the rat, and suggest CAs IV and IX as well as CAs I and II are related to electrolytes metabolism of saliva in duct cells.
Acinar Cells ; Animals ; Blotting, Western ; Carbon ; Carbonic Anhydrases ; Cytoplasm ; Electrolytes ; Immunohistochemistry ; Isoenzymes ; Membranes ; Parotid Gland ; Rats ; Saliva ; Salivary Glands ; Submandibular Gland

This study was intended to understand the role of the apoptosis-suppressing gene, bcl-2, in the hair follicle development. Immunohistochemistry for bcl-2 was performed using a high-throughput tissue-array technique, on Korean fetal scalp tissues at the 14 weeks, 16 weeks, 19 weeks & 24 weeks of the development. The results showed that the basal cells of epidermis were stained from 14 weeks to 24 weeks and the immunoreactive melanocytes were observed in the basal layer and suprabasal layer of epidermis as well as in the hair matrix cells and the external root sheath. At 19 weeks, the follicles at all stages of morphogenesis were observed. In the early stages, the epithelial cells of hair germ and hair peg, the mesenchymal cells surrounding them were stained. In the more advanced stage, the bcl-2 expression of follicular epithelial cells diminished to allow keratinization, hair canal formation and holocrine secretion to take place. In the bulbous hair peg stage, the hair papilla cells, the hair matrix cells, the basal cells of the primitive sebaceous gland, the primitive arrector pilli and the basal cells of the external root sheath were stained. We confirmed in the developing hair follicle that in the early stage or in the place where the cells continued to proliferate, the immature cells expressed bcl-2 to suppress cell death to overcome the susceptibility to cell death and when the differentiation was being achieved, the reduction of bcl-2 expression increased cell death to perform the tissue morphogenesis or the organ functions
Apoptosis ; Cell Death ; Epidermis ; Epithelial Cells ; Genes, bcl-2 ; Hair ; Hair Follicle ; Hypogonadism ; Immunohistochemistry ; Keratins ; Melanocytes ; Mitochondrial Diseases ; Morphogenesis ; Ophthalmoplegia ; Scalp ; Sebaceous Glands

Compressive neuropathy of the ulnar nerve occurs commonly at the cubital tunnel, but it can also be occurred by the anatomic variations of the structures on the ulnar nerve passage. This study was thus performed to clarify the variations of the ligaments and muscles, which can cause the ulnar nerve entrapment syndrome in the upper arm. One hundred arms of 50 Korean adult cadavers were used. The arcade of Struthers, a musculo-tendinous band from the medial head of the triceps brachii to the medial intermuscular septum, was observed in 34% of the cases. This arcade was mostly in a narrow-band shape, but a broad-band shaped arcade was sometimes observed. The internal brachial ligament was observed in 17% of cases. The epitrochleoanconeus muscle between the medial epicondyle and the olecranon was observed in 3% of cases. The ulnar nerve was wrapped or covered by the medial head of triceps brachii in 5% of cases. This study is expected to further the current understanding of the anatomic variations of ligaments and muscles on the ulnar nerve passage, and to be helpful data for the diagnosis and treatment of the ulnar nerve entrapment syndrome in the upper arm.
Adult ; Arm ; Cadaver ; Head ; Humans ; Ligaments ; Muscles ; Olecranon Process ; Ulnar Nerve ; Ulnar Nerve Compression Syndromes

The main transmitter substance mediating erection is the nitric oxide released from the vascular endothelial cells of corpus cavernosum and from the nonadrenergic, noncholinergic nerve endings. In addition, some neurotransmitters, such as acetylcholine or vasoactive intestinal polypeptide (VIP), have been reported to play an important role in mediating the erection. Thus, autonomic neuropathy may cause erectile dysfunction, and in reality, it occurs frequently in individuals with diabetes mellitus (DM), in which polyneuropathy, including both peripheral somatic sensorimotor neuropathy and autonomic neuropathy, develops usually. Thiazolidinedione (TZD) is an insulin-sensitizing agent used for the treatment of type 2 DM with insulin resistance, and has been reported to ameliorate nephropathy, decrease plasma glucose level and reduce blood pressure. However, the effect of this drug on the neuropathy related to erectile dysfunction has never been proved. In the present study, to evaluate the effect of TZDs on the neuropathy concerned with erectile dysfunction, we examined neurochemical changes of major pelvic ganglion (MPG) neurons in Otsuka Long Evans Tokushima Fatty (OLETF) rats, genetic models with non-insulin-dependent DM, after TZDs (pioglitazone and rosiglitazone) treatment. Age-matched nondiabetic Long Evans Tokushima Otsuka (LETO) rats were used as controls. Nitric oxide synthase (NOS), tyrosine hydroxylase (TH), VIP, and neuropeptide Y (NPY) contents were measured in MPG neurons of LETO, OLETF and pioglitazone- or rosiglitazone-treated OLETF rats by morphometry. Compared to the corresponding LETO group, number of TH-, NOS- and VIP-immunoreactive (ir) neurons decreased, while that of NPY-ir neurons, which modulate noradrenergic vasoconstriction of penile arteries, increased in the MPG of the OLETF group. After administration of pioglitazone- or rosiglitazone to OLETF rats for 23 weeks, these neurochemical changes were recovered to the control levels of the LETO group, although some variations were accompanied. These results suggest that TZDs treatment may be helpful for the treatment of autonomic neuropathy concerned with erectile dysfunction.
Acetylcholine ; Animals ; Arteries ; Blood Pressure ; Diabetes Mellitus ; Endothelial Cells ; Erectile Dysfunction ; Ganglion Cysts ; Glucose ; Insulin Resistance ; Male ; Models, Genetic ; Negotiating ; Nerve Endings ; Neurons ; Neuropeptide Y ; Neurotransmitter Agents ; Nitric Oxide ; Nitric Oxide Synthase ; Plasma ; Polyneuropathies ; Rats ; Rats, Inbred OLETF ; Thiazolidinediones ; Tyrosine 3-Monooxygenase ; Vasoactive Intestinal Peptide ; Vasoconstriction

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