No abstract available.
Immunohistochemistry* ; Sciuridae*

Malignant melanoma is a highly metastatic tumor, resistant to chemotherapy and radiotherapy. Recent studies have suggested that many therapeutic agents used against cancer mediate their effects by induction of apoptosis of the cancer cells. Eugenol enhances the generation of tissue-damaging free radicals and inflammation or allergic reactions. In particular, it is more cytotoxic against cancer cells compared with normal fibroblasts. This study was performed to investigate whether the cytotoxic effect of eugenol is associated with the induction of apoptosis and involves activation of caspase in the human melanoma G361 cells. Eugenol-induced apoptosis was confirmed by MTT assay, Hemacolor stain, Hoechst stain, DNA electrophoresis, and Western blot analysis. Eugenol had a significant dose- and time-dependent inhibitory effect on the viability of G361 cells. Eugenol treatment induced caspase-3 and -6 cleavage, and activation. The caspase-3 substrates PARP and DFF45 are cleaved during eugenol-induced apoptosis. It was found that the casapase-6 substrate lamin A was cleaved, whose cleavage has been reported to be necessary for complete condesation of DNA during apoptosis. These results suggest that eugenol may constitute a potential antitumor compound against melanoma occurring in the skin and oral mucosa.
Apoptosis* ; Blotting, Western ; Caspase 3 ; DNA ; Drug Therapy ; Electrophoresis ; Eugenol* ; Fibroblasts ; Free Radicals ; Humans* ; Hypersensitivity ; Inflammation ; Lamin Type A ; Melanoma* ; Mouth Mucosa ; Radiotherapy ; Skin

The purpose of this study is to evaluate the effects of Woohwangcheongsim-won on reperfusion following the MCA occlusion in rats. To evaluate the effect of Woohwangcheongsim-won on reperfusion following the MCA occlusion, the volume of cerebral ischemia and edema was measured. The volume of the control group, which was ischemic-damaged was 23.6%, and that of the sample group was 13.5%. The voluminal ratio of the right/left hemisphere was 116 in the control group, and that of the sample group was 107.
Animals ; Brain Edema ; Brain Ischemia* ; Edema ; Infarction, Middle Cerebral Artery* ; Middle Cerebral Artery* ; Neuroprotective Agents* ; Rats ; Reperfusion

The marked hemodynamic and hormonal changes of normal pregnancy are associated with striking alterations in renal physiology involving structure, dynamics, tubular function, and volume homeostasis. A number of acid-base or electrolyte disorders are associated with decreased or increased HCO3-reabsorption in the renal tubules. The present study was to examine the alterations of expression and distribution of Na+/HCO3-cotransporter (NBC), Na+/H+ exchanger-3 (NHE-3), and carbonic anhydrase I and II (CA I, II) proteins in the kidneys of non-pregnant (NP) and pregnant rats using Western blot analysis and immunohistochemistry. Sprague-Dawley female rats were studied on days 10 (P 10), 12 (P 12), 14 (P 14), 17 (P 17), and 19 (P 19) of pregnancy. Western blot analysis demonstrated that the expression of NBC, ~110 kDa at molecular mass, was increased in pregnant rats, particularly P 12, compared with NP rat. The expression of NHE-3, ~83 kDa at molecular mass, was increased in pregnant rats, particularly P 12 and P 14. The expression of CA I, ~30 kDa at molecular mass, was decreased in pregnant rats, particularly P 14, but, CA II protein, ~30 kDa at molecular mass, was similar NP rat. In immunohistochemistry, strong immunoreactivity of NBC of NP rat was exclusively detected in the basolateral membranes of S1 and S2 segment of proximal tubules whereas not in S3 segment. In pregnant rats, the pattern of cellular labeling of NBC was identical to that of NP rat, but signal intensity was increased, particularly P 12. In NHE-3, strong immunoreactivity was detected in apical membranes and brush borders of S3 segments and moderate in S1 and S2 segments. In pregnant rats, the pattern of cellular labeling was identical to that of NP rat, but the signal intensity was increased, particularly P 12 and P 14. Expression of CA I and II proteins was detected in entire collecting duct. Signal intensity was prominent in type A intercalated cells and moderate in type B intercalated cells. In pregnant rats, the pattern of cellular labeling of CA I and II proteins was identical to that of non-pregnant rat, but the signal intensity of CA I was decreased in cortical collecting duct, particularly P 14 and CA II was identical to that of NP rat. These results suggest that the regulation of NBC and NHE-3 expressions in the proximal tubules and CA I expression in cortical collecting duct may maintain HCO3-concentration during the pregnancy.
Animals ; Bicarbonates* ; Blotting, Western ; Carbonic Anhydrase I ; Female ; Hemodynamics ; Homeostasis ; Humans ; Immunohistochemistry ; Kidney* ; Membranes ; Microvilli ; Physiology ; Pregnancy ; Rats* ; Rats, Sprague-Dawley ; Social Control, Formal* ; Strikes, Employee

Injection of Freund's complete adjuvant (FCA) into the plantar surface of the rat induces inflammatory responses with accompanying pain behaviors. Signs of pain behaviors observed in FCA-injected animals are reported to be similar to symptoms seen in patients with inflammatory pain. In the previous study, injection of FCA produced a significant mechanical allodynia over time. The role of substance-P and calcitonin gene-related peptide(CGRP) on allodynia induced by inflammation is still controversial. We investigated the change of spinal neuropeptides and nitric oxide (NO) in rats with inflammation induced by subcutaneous injection of FCA into hind paw. The results are: 1. The number of NADPH-diaphorase and substance P positive neurons increased at ipsilateral spinal ventral horn after FCA injection. No significant changes were found with L-NAME posttreatment. 2. Staining intensity of substance P-immunoreactive area increased at ipsilateral spinal dorsal horn after FCA injection. No significant changes were found with L-NAME posttreatment. 3. CGRP immunoreactivity changed in the same pattern with substance P in all group. The results suggest that spinal neuropeptide substance P and CGRP are involved in the mechanism of the development and maintenance of allodynia in a state of FCA-induced inflammaion. NO may be also involved in the regulation of the quantity of substance P and CGRP in spinal cord.
Animals ; Calcitonin ; Horns ; Humans ; Hyperalgesia ; Inflammation* ; Injections, Subcutaneous ; Neurons ; Neuropeptides* ; NG-Nitroarginine Methyl Ester ; Nitric Oxide* ; Rats ; Spinal Cord ; Substance P

This study was performed in order to establish the culture system optimal for the study on pituitary prolactin cells using growth factor and extra cellular matrix components as the culture substrate. The effect of epidermal growth factor (EGF) alone or along with extracellular marix components on GH3 cell growth and PRL expression was assessed using cell count, BrdU-immunocytochemistry and PRL-immunocytochemistry in in vitro cultures on plastic, laminin and Matrigel. EGF decreased the cell growth, BrdU-labeling and increased the PRL-immunoreactive cells regardless of the culture substrate by day 3 of the culture. Matrigel was the best culture substrate to decrease the cell growth and to increase the PRL expression. EGF treatment in the Matrigel culture showed about 80.5% of PRL-immunoreactive cells by day 6 of the culture. These results indicated that Matrigel is the better culture substrate than plastic or laminin to inhibit the overgrowth and to increase the prolactin expression of the GH3 cell and that EGF and Matrigel causes very effective culture environment for the long-term culture of the GH3 cell by synergistic mechanism.
Cell Count ; Epidermal Growth Factor* ; Extracellular Matrix* ; Lactotrophs ; Laminin ; Pituitary Neoplasms* ; Plastics ; Prolactin

The cerebral artery spasm is a phenomenon frequently present in cases of subarachnoid hemorrhage (SAH). Previous reports indicated that vasospasm could be produced primarily by muscle contraction and constricted vessels that exhibit myonecrosis and subendothelial changes. Pharmacological studies have shown that vasospastic arteries decrease in contractility but increase the spontaneous tone and stiffness compared to normal cerebral arteries. We carried out electron microscopic to observe the morphological changes of the basilar artery following the experimental SAH. We examined changes in the size of the lumen and thickness of the basilar artery in SAH. Morphological changes of cerebral basilar artery following SAH : (1) The size of the lumen of basilar artery decreased about 47%, while its thickness increased about 170%. (2) The shape of the endothelial cell transformed to a cuboidal type and its thickness increased, while the length of the smooth muscle cell shortened and the thickness increased.
Arteries ; Basilar Artery* ; Cerebral Arteries ; Endothelial Cells ; Hemorrhage* ; Muscle Contraction ; Myocytes, Smooth Muscle ; Subarachnoid Hemorrhage ; Vasospasm, Intracranial

GFAP (Glial Fibrillary Acidic Protein) was one of the intermediate filament group and used as an astrocyte marker. The numerous studies about GFAP immunoreactive cell's distribution were investigated for fetus, neonate and aged brains. There are several reports about that GFAP immunoreactive cells were appeared at early fetus or after birth. In cases of mammalian fetus radial glia cells migrated toward pial surface at early stage and revealed GFAP immunoreactivity by the immunostain. But in cases of rodents, they migrated last gestation or after birth. This study, the GFAP immunoreactive cells' localizations and distribution in the fetuses (the 30th, 45th, 60th, 90th, 95th, 105th 120th of gestation) and neonate telencephalon of Korean native goat were investigated by immunohisto-chemistry (ABC method). The results obtained in this study were summarized as followings; 1. Multipolar astrocytes of 60 days of gestation were found cerebral cortex, in 95 days of gestation were found cerebral medulla, in 105 days of gestation were found lateral ventricle. 2. Radial glial cell presented 45 days of gestation and process of GFAP immunoreactive was to stretch out from ventricular to pia mater. And the nonpolar immunoreactive cells were transformed bipolar immunoreactive cells and they were transformed to monopolar and multipolar immunoreactive cell. 3. The number of GFAP immunoreactive cells of a field were gradually increased from 45 days of gestation till 90 days of gestation and decreased from 90 days of gestation till 105 days of gestation. But in 120 days of gestation and newborn were slightly increased. 4. Immunoreactivity of GFAP immunoreactive cells were gradually decreased from 95 days of gestation till 120 days of gestatioin. However, most pia mater areas and ventricles had high immunoreactivity and medulla part had low immunoreactivity. These results were suggested that radial glial cell of cerebral cortex and cerebral medulla were developed faster than lateral ventricle.
Astrocytes ; Brain ; Cerebral Cortex ; Ependymoglial Cells ; Fetus* ; Goats* ; Humans ; Immunohistochemistry ; Infant, Newborn* ; Intermediate Filaments ; Lateral Ventricles ; Neuroglia ; Parturition ; Pia Mater ; Pregnancy ; Rodentia ; Telencephalon*

We have previously demonstrated that transferrin binding protein (TfBP) is a reliable marker for mature oligoden-drocytes (OLGs) in the avian central nervous system (CNS). Unlike mammalian CNS in which OLGs are generated largely postnatally, avian OLGs are differentiated during embryonic development of CNS. In this study, several aspects of TfBP(+/-) OLG development were immunohistochemically examined in the embryonic chick cerebellum : (1) change in shapes of immature cells with respect to time and to location within the cerebellum, (2) possible sites of origin, and (3) pathways of precursor cell migration. Our results indicate that TfBP expression gradually increases and extends from the deep portion of the white matter to gray matter with proportion to progress of cerebellar development. A few TfBP? cells were first observed in the deep portion of the cerebellum at E9. At E13, TfBP(+/-) cells were distributed evenly within the white matter. At E17, many TfBP(+/-) OLGs were located at granular layer and at the near place of Purkinje cell layer. At E20, a large number of TfBP cells appeared at the granular layer with a few in the molecular layer. Our data demonstrated distinct patterns of morphology and location of TfBP(+/-) OLGs in the cerebellum during development and suggest a role of TfBP in OLG development.
Animals ; Carrier Proteins ; Cell Movement ; Central Nervous System ; Cerebellum* ; Chick Embryo ; Embryonic Development ; Female ; Oligodendroglia ; Pregnancy ; Transferrin

The presence of erythroblasts within Kupffer cell was studied for transmission electron microscopically with 5 human fetal livers from 11 to 23 weeks of gestation during the high activity of hepatic hemopoiesis. By using continuous series of thin sections electron microscopically, the objective of the present study was to evaluate the relevance between a migrated erythroblast and a Kupffer cell, and the migration of erythroblasts within Kupffer cells in the sinusoidal lumen. During the examined period the sinusoidal wall consisted of endothelial cells and Kupffer cells, being deficient in basement membrane. Erythropoietic cell-Kupffer cell interaction was often found as the emperipolesis and adhesion between the cells in human fetal liver under electron microscopy. The cytoplasm of the emperipoletic Kupffer cell contained several mitochondria, rough endoplasmic reticuli, clear vesicles, electron dense bodies, cellular debris with shrunken chromatin of enucleated nuclei, intact enucleated nuclei, and erythroblast bearing vacuoles as intact erythroblasts. Intracellular erythroblasts in the Kupffer cell remain unaltered with their normal structure and showed mitosis, enucleation and migration of erythroblast into the sinusoidal lumen. And a clear zone of a vacuole was readily seen around the intracellular erythroblast within Kupffer cell. On occasion, the hypertropic Kupffer cell with interacellular erythroblasts virtually occluded the sinusoidal lining cell. Processing of a migrating emperipoletic erythroblast within a Kupffer cell, the erythroblast migrated via migration pore through the luminal cell membrane of the Kupffer cell into the sinusoidal lumen. An invasion of a proerythroblast into Kupffer cell or a migration of the cell into the sinusoidal lumen had been found in human fetal liver from 11 to 13 weeks of gestation. The results demonstrate that migration of emperipoletic erythroblasts within Kupffer cells occurs in human hepatic hemopoiesis. We suggest that emperipolsis may be one of the mechanisms that support the maturation of erythroblasts in human fetal liver.
Basement Membrane ; Cell Communication ; Cell Membrane ; Chromatin ; Cytoplasm ; Emperipolesis ; Endothelial Cells ; Erythroblasts* ; Humans* ; Kupffer Cells* ; Liver ; Microscopy, Electron ; Mitochondria ; Mitosis ; Phenobarbital ; Pregnancy ; Vacuoles