Distribution of von Willebrand Factor with Angiogenesis in Neonatal Rat Heart / 체질인류학회지

To investigate the process of angiogenesis in the cardiac tissue of neonatal rats, the levels of von Willebrand factor (vWf) produced by endothelial cells were observed. At days 1 to 7 after birth, whole heart obtained from neonatal rats was frozen sectioned, stained with anti-vWf and biotinylated rabbit anti-goat IgG antibodies, followed by immunohistochemical examinations. The results were as follows: 1. At day 1 after birth, extracellular matrix of endocardium and epicardium was stained with anti-vWf at the intermediate level, but that of myocardium was at the low level. 2. At day 2 after birth, a few blood islands were detected. At day 4 after birth, blood island was formed in most parts of heart and extracellular matrix was stained with ant-vWf at the intermediate level. 3. At day 7 after birth, a few blood vessels were formed, and endothelial cells and extracellular matrix was stained with ant-vWf at the intermediate level. These results suggest that mesenchymal cells were differentiated to blood islands and myocardiac cells, which are responsible for the distribution of vWf in extracellular matirx and for angiogenesis.

Immunohistochemical and Electron Microscopic Study on the Depigmentation Effect of Melanoston / 체질인류학회지

The present study was designed to address whether melanoston which was extracted from yeast was able to induce depigmentation of the skin in guinea pigs. The activity of tyrosinase which participates in the synthesis of melanin pigment residing in melanocytes of guinea pig's epidermis was assessed using immunohistochemical staining methods with anti-tyrosinase antibody. To determine whether melanoston affects the processes of melanin synthesis in melanosome of melanocytes, melanoston-applied skin was examined by electron microscopy following immunogold tagging methods. We obtained results as follows: 1. The activity of tyrosinase was the lowest 3 weeks after application of 0.1 and 0.01% melanoston on the skin of guinea pigs. 2. The frequency of immunogold particles distributed in melanocytes and keratinocytes was lowest 3-4 weeks after application of 0.5 and 0.1% melanoston. Thus, these results suggest that melanoston has a potential to inhibit synthesis and distribution of melanin in the epidermis.

Essential roles for ID-1 motif of interleukin-4 receptor alpha chain in interleukin-4 signaling / 천식및알레르기

BACKGROUND: Interleukin (IL)-4 is a pleiotropic cytokine that plays an important role in the pathogenesis of the allergic inflammation and asthma. Upon IL-4 receptor (IL-4R) engagement, a variety of signaling mediators, such as JAK kinases and STAT-6 are activated, leading to induction of IL-4 target gene expression including CD23 and germline C epsilon transcription. The function of a membrane-proximal domain of IL-4Ra, termed ID-1, remains to be characterized to date. OBJECTIVE: To assess whether the ID-1 domain mediates the induction of IL-4 target gene expression in a STAT-6-dependent manner. METHODS: The intracellular region of IL-4Ralpha was translationally fused to the extracellular region of IL-2Rbeta to provide ligand specificity to IL-2. Acidic amino acids and serine residues in the ID-1 domain of the chimeric receptor were substituted by site-directed mutagenesis. These receptor cDNAs were stably transfected to M12.4.1 murine B lymphoma cells. Following IL-2 stimulation, wild type and mutant clones for the ID-1 motif were subjected to FACS. RNA blotting and elecroporetic mobility shift assays to address the levels of CD23, germline C epsilon and STAT-6 inductions, respectively. RESULTS: ID-1 mutant clones were defective in gene induction of CD23 and germline C epsilon in response to IL-2 stimulation, as compared with wildtype clones. Moreover, IL-2-mediated STAT-6 activation was abolished in ID-1 mutant clones. CONCLUSION: These results demonstrate that the ID-1 domain of IL-4Ra is essential to induce IL-4 target gene expression through a STAT-6-dependent pathway.

Changes in Laminin beta 1 Chain Expression in Renal Cortex by the Intervention with Cilazapril and Losartan on Adriamycin-Induced Nephropathy / 대한해부학회지

Laminin, an extracellular matrix glycoprotein, is distributed in the basement membrane of renal glomerulus. Laminin has been demonstrated to regulate the development and differentiation of glomeruli and play an important role in the filtering function of glomeruli. To address whether laminin beta 1 chain is associated with the pathogenic states of renal injury and recovery, nephropathy were induced by intravenous adriamycin injection and subsequently cured by treatment with antihypertensive drugs, cilazapril (angiotensin-converting enzyme inhibitor, ACEi) and lozartan (angiotensin receptor blockade, ARB), which have been known to be effective on renal protection. The results we obtained were as follows: 1. At 6 and 12 weeks after adriamycin injection, epithelial cells of proximal convoluted tubules exhibited intensive deposition of laminin beta 1 chain reactive gold particles. Cytoplasm of podocytes and mesangial cells in glomeruli showed an increase in the gold particle deposition as compared with that of controls. The mRNA level of laminin beta 1 chain was enhanced in the epithelial cells of proximal convoluted tubules. 2. Following cilazapril administration to adriamycin -induced nephropathy rats, laminin beta 1 chain reactive gold particle deposition was reduced in the epithelial cells of proximal convoluted tubules, podocytes, and mesangial cells. The level of laminin beta 1 chain transcripts was remarkably decreased in the epithelial cells of proximal convoluted tubules. 3. Following losartan administration to adriamycin -induced nephropathy rats, podocytes and mesangial cells were deposited with reduced number of laminin beta 1 chain reactive gold particles. The laminin beta 1 chain reactive gold particles and level of laminin beta 1 chain mRNA transcripts were remarkably reduced in the epithelial cells of proximal convoluted tubules. 4. Treatment with mixture of cilazapril and losartan to adriamycin -induced nephropathy rat resulted in a decrease in laminin beta 1 chain reactive gold particle deposition within the epithelial cells of proximal convoluted tubules, podocytes, and mesangial cells. Laminin beta 1 chain mRNA expression in the epithelial cells of proximal convoluted tubules nearly disappeared. These results consequently suggest that laminin beta 1 chain may be synthesized within the epithelial cells of proximal convoluted tubules, podocytes and mesangial cells and laminin beta 1 chain may play an important role on tissue recovery of the renal tissues injured by adriamycin.

The Influence of Adriamycin on the Morphologic Alteration and the Expression of Laminin in the Renal Glomerular Basement Membrane of Rat / 대한신장학회잡지

Laminin is a glycoprotein that is composed of the basement membrane of renal glomerulus and various tissues, such as epithelium, nervous tissue, and muscle tissues. Functions of laminin have been demonstrated to play an essential role in the regeneration and polarity of cells as well as the reservation of materials inside of the tissues. In collaboration with type IV collagen, heparan sulfate proteoglycan, and fibronectin, laminin forms a spike-like structure in the renal glomerular basement membrane of normal kidney. Recently, many investigators have suggested that the distribution of laminin in the basement membrane was altered in a variety of renal diseases, expecially in membranous lupus nephritis, membranous proliferative gromerular nephritis, and glomerulosclerosis. To investigate whether the morphologic changes in the glomerulus are associated with the profile of laminin distribution in the renal glomerular basement membrane, the gromerulus was examined after exposure to adriamycin, a potent inducer of nephritis. Male rats(Sprague Dawley strain) were intraperitoneally injected with 0.2ml adriamycin at 25mg/kg body weight. At 24 hour, 48 hour, 72 hour, one week, and 2 week after adriamycin administration, kidney isolated from sacrificed rats was sectioned and observed by immunohistochemical staining and electron microscopy using the immunogold method. The results we obtained were as follows: 1) Renal glomerulus was strongly stained with anti-laminin antibody when 24 or 48 hours passed after adriamycin administration. However, after 1 or 2 weeks later, the antibody response became weak at the level comparable with the vehicle controls. The renal glomerular basement membrane exhibited an increase of gold particles in subjects passed both 24 and 48 hours after adriamycin administration. This increase was observed in mesangial cells in the same subjects. In contrast, both 1 and 2 week-passed subjects displayed a reduction in the binding of gold particles at the level comparable with the vehicle controls. 2) 24 hour-passed rats upon treatment with adriamycin showed various morphologic changes in the renal glomerulus, including thicker and less transparent basement membrane and undetectable lamina densa layer. In conclusion, these results suggest that adriamycin-induced renal toxicity leads to the morphologic alteration along with an increase in laminin expression and that these aspects may be attributable to the regenerative potential of laminin.

Electron Microscopic Studies on the Effects of Retinoic Acid on Palatogenesis in Fetal Rats / 체질인류학회지

Retinoic acid (RA) is widely used to treat the dermatologic disorders, such as acne and psoriasis, but its usage is limited because of teratogenic effects. Moreover, it is known that RA induces cleft palate by influencing epithelial differentiation and mesenchymal cells in palatine processes. We studied the ultrastructures of the epithelial and mesenchymal cells in rat palatine shelves treated with RA, in comparison with those of the normal developing rat. In this experiment, pregnant Sprague -Dawley rats were treated with 100 mg/kg of all -trans retinoic acid at day 10 of gestation. Pregnant rats were killed at 14 th and 16 th day of gestation. Fetuses were removed and palatine processes were dissected. The specimen were observed with a transmissiom electron microscope. The results were as follows. 1. Palatine epithelium of control rats was made up of two cell layers at day 14 of gestation, and that of RA treated rats consisted of multicellular layers. At the 16th day of gestation, many apoptotic bodies were observed in triangular area of the palatine epithelium of the control rat. In contrast, apoptotic cells were hardly observed in RA treated rats. 2. Mesenchymal cells of control rats contained cytoplasmic process, oval -shaped nucleus, well -developed rough endoplasmic reticulum, Golgi complex, and mitochondria. RA treated mesenchymal cells showed atrophied cisternae of Golgi complex, rough endoplasmic reticulum with sacculated, fragmented and ribosome detached cisternae, mitochondria with dissolved mitochondrial cristae, and multivesicular body. After RA exposure during palatogenesis, the frequency of apoptotic bodies was low in palatine epithelium, and mesenchymal cells were severely damaged. In conclusion, it is suggested the RA may induce direct cytotoxic effects on mesenchymal cells and influence normal apoptosis process in developing epithelium.

Selective Susceptibility of Synovial Fibroblasts Isolated from Patients with Rheumatoid Arthritis to TRAIL -induced Cell Death / 체질인류학회지

TNF -related apoptosis inducing ligand (TRAIL) is a member of TNF ligand superfamily. TRAIL transduces death signal through two distinct receptors, TRAILR -1I and TRAILR -2, while the engagement of TRAILR -3 and TRAILR -4 interferes with TRAIL -induced apoptosis. The profile of TRAILR expression has been reported to be a mechanism by which transformed cells undergo apoptosis in response to TRAIL while normal cells do not. Rheumatoid arthritis (RA) is an inflammatory autoimmune disease which is characterized by the hyperplasia of synovial membrane. The dysregulation of apoptosis in synoviocytes has been suggested to contribute to synovial hyperplasia. Synovial fibroblasts obtained from patients with RA have been reported to exhibit several semi - transformed aspects. To investigate whether RA synovial fibroblasts acquire the susceptibility to TRAIL -induced apoptosis, synovial fibroblast lines obtained from 2 RA patients and two osteoarthritis (OA) patients were cultured in the presence of recombinant human TRAIL and followed by MTT assay. TRAIL treatment resulted in a significant decrease in the viability of both lines of RA cells, indicating TRAIL -induced cell death of RA synovial fibroblasts, whereas OA synovial fibroblasts and normal human dermal fibroblasts were either resistant or less sensitive to TRAIL as compared with RA synovial fibroblasts. In RT -PCR analyses, the expression levels of TRAILR 4 in RA synovial fibroblasts were lower than in OA synovial fibroblasts, while other receptors in both cell lines were expressed at comparable levels. Immunohistochemical studies showed that in RA synovial tissues TRAILR -3cells were mainly leukocyte infiltrates, implying that such leukocyte infiltrates play a role in the perpetuation of the disease. Taken together, these results suggest that RA synovial fibroblasts acquire the susceptibility to TRAIL -induced cell death during disease progression and this death signal may be regulated by, at least in part, differential expression of TRAILR -4 molecule.

Localization of Laminin and Laminin beta 1 chain on Gomeruli of Developmental Rat Kidney / 체질인류학회지

Laminin, an extracellular matrix glycoprotein composed of three polypeptide chains such as alpha , beta, and gamma is distributed in basement membranes of epithelium, muscle, and nervous tissues. Laminin functions as an extracellular cytoskeleton and regulates the differentiation and polarization of cells adjacent to the basement membrane. Along with type IV collagen and heparan sulfate proteoglycan, laminin forms a spike -like structure in the renal glomerular basement membrane (GBM). It has been previously demonstrated that the distribution and immune reaction of laminin are changed in response to the conditions of glomerulonephritis and that laminin plays a role in the reformation of GBM as well as the regeneration of renal glomerular cells. In the present study, the profile of expression and distribution of laminin/laminin beta1 chain were examined in different developmental stages and upon adriamycin administration. Kidney obtained from fetuses (16, 18, and 20 days old) and infants (1 and 7 days old) of Sprague -Dawley rats were either cryosectioned for immunohistochemical assays or ultrathin -sectioned for electron microscopy using immunogold staining methods. The results were as follows: 1. Intensive expression of laminin was observed in the GBM and surrounding mesenchymal tissues obtained from 16, 18, and 20 days old fetuses and in the glomerulus from one day neonates, whereas the level of staining decreased in the glomerulus from 7 days old infants. 2. Immunogold particles were observed in the comma -shaped nephron, in particular in cisternae of rough endoplasmic reticulum, vesicles and nuclear membrane of endothelial cells and mesangial cells obtained from 18 days old fetuses. 3. The immune reactions of laminin beta1 chain were trace detected in the kidney from fetuses (16, 18, and 20 days old) and weakly in tissues surrounding blood capillary and mesangial tissues from one day old neonates. 4. After 24 hours following adriamycin treatment, the reactivity of laminin was slightly enhanced in the renal glomerulus, when compared with that of untreated controls. This enhancement persisted up to 1 week of adriamycin treatment. Laminin beta1 chain was weakly detectable, while further treatment with adriamycin for another 24 hours reduced the intensity of laminin beta1 chain. Taken together, these results suggest that laminin is localized in the GBM at the high level during early fetal stages but the expression levels decrease after birth. Moreover, administration with adriamycin may result in an increase in the immune reactivities of laminin and laminin beta1 chain by renal tissue damage followed by renal regeneration.

Protective Roles of Adenosine A1 Receptor Agonist and KATP Channel Opener in Adriamycin induced Cardiac Toxicity / 체질인류학회지

Although adriamycin is a potent chemotherapeutic agent, it elicits serious adverse effects, including cardiac toxicity. Evidence suggests that congestive heart failure induced by adriamycin is mediated by oxidative stress. We investigated whether regulators of adenosine A1 receptor and KATP channel, which have been demonstrated to mediate protective effects of ischemic -preconditioning in myocardium, are able to modulate adriamicin -induced impairment of cardiomyocyte. To study the effect of antioxidant, adenosine A1 receptor agonist & antagonist and KATP channel agonist & antagonist, ICR mice were pretreated with Cu,Zn -SOD, dimethyl thiourea, RPIA (R (-)N6 -(2 -Phenylisopropropyl)- adenosine, adenosine A1 receptor agonist), 8 -CPDPX (8 -Cyclopentyl -1, 3 -dipropylxanthine, adenosine A1 receptor antagonist), Pinacidil (KATP channel opener) and glibenclamide (KATP channel closer), followed by i.p injection with adriamycin. Mice were sacrificed day 1 or day 4 after adriamycin injection and cardiac toxicity was accessed by measurement of creatine phosphokinase (CK) levels in serum, immunohistochemistry using anti -Bcl -2 antibody and TUNEL histochemical assay. As expected, pretreatment of mice with Cu, Zn -SOD and DMTU reduced the frequency of TUNEL positive cells, indicating antioxidants protected cardiocytes from adriamycin -induced apoptosis. Interestingly, pretreatment with RPIA and pinacidil induced a significant decrease in adriamycin -induced cytotoxicity, whereas 8 -CPDPX and glibenclamide generated the opposite results. In Bcl -2 immunohistochemistry, an increased expression of Bcl -2 was found in all ADR treated groups, especially in glibenclamide pretreated group, and 8 -CPDPX pretreated groups, but Bcl -2 failed to protect myocytes from apoptosis. All ADR treated groups exhibited elevated levels of serum CK, compared with nomal controls, especially mice sacrificed at day 4 than those at day 1, and showed similar patterns of TUNNEL assay, reflecting heart tissue damages. This observation implicated cytoprotective roles of RPIA and pinacidil against adriamycin -induced cardiac toxicity. In conclusion, these results demonstrated that adriamycin -induced cardiotoxicity was associated with the generation of reactive oxygen species and that regulators including SOD, DMTU, RPIA and pinacidil elicited protective effects on this toxicity. In particular, pinacidil, the KATP channel opener, was more effective than RPIA, the adenosine A, receptor agonist, to attenate the adriamycin -induced cardiac toxicity.

The Effects of Retinoic Acid on Development of the Knee Joint in Rat / 체질인류학회지

No abstract available.

Effects of Ischemic Preconditioning, Adenosine and Pinacidil on the Expression of Cu,Zn- and Mn-SOD mRNA in the Rectus Femoris Muscle of the Rat after Ischemia and Timely Reperfusion / 체질인류학회지

A brief episode of ischemia and reperfusion termed 'ischemic preconditioning' has been established as rendering muscle tolerance to damage during a subsequent prolonged ischemia. The effects of ischemic preconditioning in the cardiac muscle are related to the stimulation of adenosine A1 receptor and the opening of KATP channel. The effect and mechanisms of ischemic preconditioning in the skeletal muscle are not known clearly. The superoxide radical injures the skeletal muscle during the ischemia and reperfusion. There are two types of SOD, which metabolizes the superoxide radicals to H2O2 and O2, in the cell. One of them is Cu, Zn-SOD in the cytoplasm and the other is Mn-SOD in the mitochondria. The activities of SOD are increased against the formation of superoxide radical during the reperfusion. The author performed the present study to investigate the effect and the mechanisms of ischemic preconditioning by measuring the expression of SOD mRNA on timely reperfused ischemic muscles. The healthy Sprague-Dawley rats weighing from 300 g to 350 g were used as experimental animals. Under pentobarbital (50 mg/kg) anesthesia, lower abdominal incision was done and left common iliac artery was occluded by vascular clamp for 2 hours. Rectus femoris muscles were obtained respectively at 3, 6, 12, 24 and 72 hours after reperfusion. The ischemic preconditioning group underwent three episodes of 5 minute occlusion and 5 minute reperfusion of common iliac artery followed by 2 hours of ischemia and timely reperfusion. Adenosine (50 microgram/kg) or pinacidil (1 mg/kg) was administered intravenously before ischemia. 8-cyclopentyl-1, 3-dipropylxanthine (15 mg/kg) or glibenclamide (0.5 mg/kg) was administered intravenously before ischemic preconditioning. Paraffin sections with 4 micrometer thickness in all groups were obtained. The expression of Cu, Zn- and Mn-SOD mRNA was observed by use of in situ hybridization. The results obtained were as follows. 1. The expression of SOD mRNA was seen only in small muscle fibers of the rectus femoris muscle of the rat. 2. Weak expressions of Cu, Zn- and Mn-SOD mRNA were observed in the normal control rat. 3. After 2 hours of ischemia, moderate expression of Cu, Zn-SOD mRNA was observed until 72 hours of reperfusion. Weak or moderate expression of Mn-SOD mRNA at 3 hours and 6 hours of reperfusion, weak or trace expression at 12 hours of reperfusion, moderate expression at 24 hours of reperfusion and weak or moderate expression at 72 hours of reperfusion were observed. 4. After ischemic preconditioning, moderate expressions of Cu, Zn-SOD mRNA were seen in the groups of 3, 6, 12 and 24 hours of reperfusion. Moderate expressions of Mn-SOD mRNA were seen in the group of 0, 3, 6 and 12 hours of reperfusion and strong expression was seen in the group of 24 hours of reperfusion after ischemic preconditioning. 5. After 2 hours of ischemia with ischemic preconditoining, moderate expressions of Cu, Zn-SOD mRNA were seen in the groups of 0, 3, 6, 12, 24 hours of reperfusion. Moderate expressions of Mn-SOD mRNA were observed in the groups of 0, 3, 6, and 12 hours of reperfusion and moderate or strong expression was seen in the group of 24 hours of reperfusion. 6. After 2 hours of ischemia with the pretreatment of adenosine, moderate expressions of Cu, Zn-SOD mRNA were seen in the group of 0, 3, 6, 12 and 24 hours of reperfusion. Moderate expression of Mn-SOD mRNA in the groups and 3 hours of reperfusion, strong expression in the group of 6 and 12 hours of reperfusion and moderate expression in the group of 24 hours of reperfusion were seen. 7. After 2 hours of ischemia with the pretreatment of pinacidil, moderate expressions of Cu, Zn-SOD mRNA were seen in the groups of 0, 3, 6 and 12 hours of reperfusion and those of Mn-SOD mRNA were seen in the groups of 3, 6, 12 and 24 hours of reperfusion. 8. After 2 hours of ischemia with ischemic preconditioning and the pretreatment of 8-cyclopentyl-1, 3- dipropylxanthine, moderate expression of Cu, Zn-SOD mRNA were observed in the groups of 0, 3, 6, and 12 hours of reperfusion and those of Mn-SOD were seen in the groups of 6, 12 and 72 hours of reperfusion. 9. After 2 hours of ischemia with ischemic preconditioning and the pretreatment of glibenclamide, moderate expressions of Cu, Zn- and Mn-SOD mRNA were seen in all groups of reperfusion. Consequently, these results suggest that the expression of Cu, Zn and Mn-SOD mRNA increases during 2 hours ischemia and reperfusion with or without ischemic preconditioning. The effects of ischemic preconditioning are closely related to the stimulation of adenosine A1 receptor and KATP channel.

Expression and Localization of Laminin on Rectus Femoris Muscle in the Developing Rats / 대한해부학회지

Laminin is an extracellular matrix-associated protein, which is largely present in the basement membranes of the human placenta, striated muscle and Schwann cell. Laminin has been proposed to promote attachment, spread, motility, growth and development of tissues or cells. In this study, we investigated the formation, localization and migration of the laminin in developing rectus femoris muscle of fetal and newborn rats. Experimental animals, fetal or newborn rats (Sprague-Dawley strain), were divided into 8 groups (14 day-old, 16 day-old, 18 day-old, 20 day-old and 22 day-old fetal rats, 1 day-old, 3 day-old, 5 day-old and 7 day-old newborn rats). The specimens of each group were prepared for detection of the laminin by immuno- histological and immunogold electron microscopic methods. 1. The primitive rectus femoris muscle of 14 day-old fetal rats consisted of mesenchymal cells and myoblasts. Laminin stained with a few gold particles were observed in the cytoplasm of myoblasts and rough endoplasmic reticulum of mesenchymal cells. 2. In the rectus femoris muscle of 16 day-old fetal rats, laminin was strongly expressed in myoblasts and mesenchymal cells. Gold particles were distributed in the cytoplasm of myoblasts and mesenchymal cells. 3. In the rectus femoris muscle of 18 and 20 day-old fetal rats, strong laminin immune reactions were observed in the basement membrane of muscle cells. A few gold particles distributed on the basal lamina of muscle cells and in the RER of fibroblasts. 4. In the rectus femoris muscle of 22 day-old fetal rats and 1 day-old newborn rats, strong laminin immune reactions were seen in the basement membrane of muscle cells. Numerous gold particles were located on the basal lamina of muscle cells and the RER of fibroblasts. 5. In the rectus femoris muscle of 3 day, 5 day and 7 day-old newborn rats, moderate laminin immune reactions were detected in the basement membrane of the muscle cell. A few gold particles were located on the basal laminae of muscle cells and RER of fibroblasts. These results suggest that the distribution of laminin in the rectus femoris muscle is related to myogenicity. Laminin seemed to be secreted by myoblasts of rectus femoris muscle at the fetal stage, but by fibroblasts in the muscle at neonatal and adult stages.

N-cadherin Immunoreaction in the Interventricular Septum in Developing Rat Heart

BACKGROUND AND OBJECTIVES: N-cadherin is known to be expressed in neuroectodermal tissue such as central nervous system and various mesodermal origin tissues such as kidney and heart. We investigated N-cadherin expression in the endocardial cushion in developing rat heart by immunohistochemical method. MATERIALS AND METHOD: Fetal rat hearts at the 11th, 13th, 15th, 17th, and 19th day of gestation and the 1st day neonatal rat heart were used. Hematoxylin and eosin stain was performed for normal cardiogenesis, and immunohistochemistry was performed for the expression of N-cadherin in interventricular septum(IVS) during cardiogenesis in rat. RESULTS: Ventricular wall and membranous part of the IVS showed positive reaction with anti-N-cadherin at the 11th day of gestation. Membranous part of IVS was begun to show tracely positive reaction at the 15th day of gestation, and thereafter the immunoreactivity was increased with maturation. At the 17th day of gestation mesenchymal cells in membranous muscular part of the IVS showed positive reaction. The similar immunoreactivity of membranous and muscular parts of IVS were shown at the 19th day of gestation. CONCLUSION: As the immunoreaction of mesenchymal cells in the membraneous part of IVS to anti-N-cadherin was increased with time, it is suggested that mesenchymal cells in membranous part of IVS were differentiated into the cardiomyocytes.

Development of the Shoulder Joint in Staged Human Embryos and Fetuses in Korean / 체질인류학회지

The authors have studied 37 Korean embryos of Carnegie stage 11~23 and 18 fetuses to demonstrate the development of the shoulder joint. The external feature of the upperlimb bud is observed by stereoscope and camera, and the internal structures are studied by microscopic observation. The results obtained were as follows: In stage 12 upperlimb buds were appeared. In stage 17 mesenchymal condensations for humerus and scapula, and glenoid labrum were observed. In stage 19 one-layered interzone between the humerus and scapula was visible. In stage 22 three-layered interzone between the humerus and glenoid labrum was formed. In stage 23 three-layered interzone between the humerus and glenoid fossa of scapula was visible. In the 9th and 10th weeks distinct joint cavity was formed between humerus and glenoid labrum, and tendon of long head of Biceps brachii was attached to supraglenoid tuberosity and glenoid labrum. In the 11th week the joint cavity was formed between the midportion of humerus and glenoid fossa, and tendon of long head of Biceps brachii was more dense. In the 16th week the glenoid labrum was visible as fibrous cartilage, and joint cavity was more widened. In the 20th to 32nd week the shoulder joint was matured with the distinct joint cavity and glenoid labrum time after time.

Ultrastructural Changes in Rat Anterior Tibial and Soleus Muscles Following Graded Periods of Ischemia and Reperfusion / 체질인류학회지

Skeletal muscles are known to have tolerance to ischemia, but a prolonged ischemia can cause damage to muscular tissues. The ischemia-reperfusion injury results from the oxygen free radicals released by leucocytes and formed by the reaction of hypoxanthine and xanthine oxidase. Superoxide dismutase (SOD), one of major antioxidant enzymes ocurring in the various tissues of the body metabolizes or scarvanges the oxygen free radicals. Although many studies reported difference in tolerance to ischemia and reperfusion between white and red muscles, some other investigators failed in finding such difference. The present study was performed to examine effects of graded periods of ischemia and reperfusion on the cellular ultrastructure and activity of SOD in white and red muscles. The Sprague-Dawley rats (200~250 g) were used as experimental animals. Under pentobarbital (50 mg/kg IP) anesthesia, incision was made on lower abdomen and left common iliac artery was occluded by means of a vascular clamp for 2, 4 and 6 hour (hrs). Thereafter, the superficial portion of mid-belly of anterior tibial muscle and soleus muscles were excised at 0, 24 and 72 hrs after onset of reperfusion. The specimens were sectioned into slices, 2 mm in length, 1 mm in width and thickness. Some specimens were prepared for electron microscopic observation and others for determination of SOD activity by using antihuman Cu, Zn- and Mn-SOD antibodies. The results obtained were as follows. 1. In anterior tibial muscle, areas with loose electron-density and dilated cristae were observed in the mitochondria immediately after 2 hrs of ischemia, while widened intermyofibrillar spaces and dilated cisternae of sarcoplasmic reticulum were seen after 2 hrs and 24 hrs reperfusion. When subjected to 2 hrs ischemia and 72 hrs reperfusion, no significant change was found in the cellular ultrastructure. 2. In soleus muscle, electron density was loose in the matrix of mitochondria immediately after 2 hrs of ischemia, while cisternae of sarcoplasmic reticulum were dilalated after 2 hrs of ischemia and 24 hrs reperfusion. Following 2 hrs of ischemia and 72 hrs reperfusion, the electron microscopic findings were similar to those of normal rats. 3. The changes in cellular ultrastructure were more prominent in both the 4 hrs and 6 hrs ischemia groups, in which degree of ultrastructural changes were proportional to duration of reperfusion. 4. In anterior tibial muscle, trace or weak immunoreactivities of Cu, Zn- and Mn-SOD were seen, whereas trace immunoreactivity of Cu, Zn-SOD and trace or weak immunoreactivity of Mn-SOD were observed in soleus muscle. 5. The immunoreactivities of Cu, Zn- and Mn-SOD were not altered in 2 hrs ischemic and 72 hrs reperfused group, while they were increased slightly in 2 hrs ischemic and 24 hrs reperfused group. 6. In both muscles, the activity of SOD increased following 4 hrs or 6 hrs ischemia and 24 hrs or 72 hrs reperfusion. The changes in immunoreactivity of Mn-SOD were not different between two muscles, whereas immunoreactivity of Cu, Zn-SOD were higher in anterior tibial muscle. Consequently, it is suggested that significant ischemia reperfusion injuries are produced after 4~6 hrs ishemia followed by 24 hrs or 72 hrs reperfusion, that anterior tibial muscle is more susceptible to ischemic reperfusion injury and that the ischemic-reperfusion injury is closely related with activity of SOD.

Immunohistological and Immunogold Electronmicroscopic Studies on Fibronectin in Lung and Liver of CS gas Exposed Rats / 체질인류학회지

Fibronectin (FN) is a major extracellular matrix glycoprotein, highly expressed in developing rat lungs. Several observations suggest that it play an important role in many developmental processes of animals. In vitro, FN can affect the adhesion, migration, proliferation, differentiation, and even apoptosis of various cell types. This study was undertaken to describe the distribution and localizations of fibronectin in the alveolar septum of lungs and liver lobules after CS gas exposure to experimental rats. The experimental rats (Sprague-Dawley strain), weighing 150~200 gm were sacrificed at 6 hours, 12 hours, 24 hours, 48 hours and 72 hours after CS gas exposure. The specimens of lung and liver were prepared for fibronectin immunoreactions of alveolar septa and liver lobules on experimental group, and for fibronectin reactions on the cytoplasm of type I alveolar cells, type II alveolar cells, fibroblasts, alveolar macrophages and interstitium in alveolar septa of lungs. All of specimens for immune reactions were observed with light and electron microscopes. The results obtained were as follows. 1. The liver lobules showed mild fibronectin reactions at the 6 hours and 12 hours after CS gas exposure. 2. The alveolar macrophages revealed strong fibronectin reactions. But alveolar septa showed weak FN reactions at 6 hours after CS gas exposure. 3. At 12 hours and 24 hours after CS gas exposure, the interstitial pneumonitis were seen in the lung alveoli. The gold particles were increased in the cells of alveolar septa, weak fibronectin reactions were revealed in the alveolar septum. 4. At 48 hours and 72 hours after CS gas exposure, FN reactions of alveolar septa were moderate, and the gold particles in the alveolar cells were markedly decreased. These results suggest that CS gas exposure to rats induces the increase of the fibronectin in lung and liver.

The Effect of Uterine Environment during Peri-implantation Period on theUltrastructure of Zona Pellucida in Mouse Oocytes and Embryos / 대한불임학회지

In the studies on the hatching mechanisms in mammals, many investigators focused on the embryonic intrinsic factor(s) in in vitro culture, but the uterine environment as the extrinsic factor(s) is thought to play an important role in hatching mechanism. Therefore, to evaluate the effect of uterine environment on the hatching event in vivo, the immature(GV) and ovulated(MII) oocytes, and the late 2-cell embryos of mouse were transferred to pseudopregnant foster mother's uterus during peri-implantation period. So it was verified whether there would happen hatching by only uterine environment independently on embryonic stage. The ultrastructural changes of the zona surface of transferred group were compared with those of in vivo and vitro group by SEM. 36 hrs after transfer, the immature and ovulated oocytes almost degenerated, and the late 2-cell embryos developed to various embryonic stages. However, the embryos which didn't develop to blastula stage did not hatch. The ultrastructural network of ZP in transferred group seemed to be smoothed uniformly, which was different from in vitro group. In conclusion, it is suggested that the uterine environment during peri-implantation period enhances the embryo hatching by provoking the structural change of ZP.

The Distribution Patterns of the Cutaneous Nerves on Dorsum of the Hands in the Korean / 체질인류학회지

In order to investigate the distribution patterns of the dorsal digital nerves of the radial and ulnar nerve in the Korean, authors dissect the 113 hands (right 58/left 55) of the 59 cadavers (39 males/20 females). The types were classified by the area of radial dorsal digital nerves and the ulnar digital nerves. The difference in the distribution pattern between males and females, right and left hands was analysed by chi2-test in the case presenting the prequency more than 10%. The results as follows; 1. The ten types of the distribution patterns consisted of the radial and ulnar nerves were observed on the dorsum of the hands. 2. The case of the highest prequency was type VIII(33.9%), in which radial nerve supply the radial side of the 2 1-2 of digits and ulnar nerve extends the ulnar side 2 1-2 of digits. 3. In the cases of the both nerve mingling in the third digital web, the incidences in which the radial nerves extend to the radial half of ring finger and ulnar nerve to the ulnar half of middle finger (type III) were 25.7%, and that the radial nerves extend to the ulnar half of middle finger and ulnar nerve to the ulnar half of middle finger (type VI) were 11.0%. 4. Type IV as combined branch between the radial and the ulnar nerve extend to the third digital web was observed in the 12.8%. 5. The type III, VI, IX, X, XI showing the both nerves mingling in the third digital web and in the second digital web or combining in the second digital web were new observed in the Korean. 6. The musculocutaneous nerve replaces the superficial branch of the radial nerve in 4 cases. 7. There was no statistical difference in the distribution pattern between males and females, right and left hands. From the above results, it was suggested that the majority of the cases were that the ulnar digital nerves supplied the ulnar half of the middle finger in the Korean.

Effect of DMTU Pretreatment on Ultrastructural Changes in Tibialis Anterior Muscles and Soleus Muscles of Rats after Irradiation / 체질인류학회지

Free radicals, formed by ionization of water molecules, cause significant increase of morbidity and mortality in irradiated humans. The skeletal muscle is relatively radio-resistant because of its few content of proliferating cells. But the incidence and severity of muscular damage depends on the dose of radiation and time lapse. This study is aimed to investigate the ultrastructural changes and the effect of radiation and DMTU on two muscles, the tibialis anterior and the soleus muscle; the former is dominantly composed of white muscles while the latter is mainly composed of red muscles. Each muscle also show differences in energy production and distribution of capillaries. The male Sprague-Dawley rats weighing from 200 g to 250 g were used as experimental animals. Under urethane(1.15 g/kg, i.p.) anesthesia, 30Gy irradiation to lower extremities with PICKER-C9 cobalt 60 teletherapy unit was done. DMTU(500 mg/kg, i.p.) was administered 1 hour prior to irradiation. The animals were sacrificed 1 day, 3 days, 7 days, 2 weeks, and 4 weeks after irradiation. The muscular tissues in midbelly of tibialis anterior and the soleus muscles were obtained and sliced into 2 mm in length, 1 mm in width and thickness. The specimens were prepared by routine method for electron microscopy. The results obtained were as follows: 1. Widening of interfibrillar space, mitochondrial changes of eletron-lucent matrix and dilated cristae, and cisternal dilatation of sarcoplasmic reticulum were observed in both muscles after irradiation. More severe ultrastructural changes with time course were observed by 2 weeks. But those were recovered to normal at 4 weeks after irradiation. 2. More severe ultrastructural changes in soleus were observed 1 and 3 days after irradiation, and in tibialis anterior at 7 days and 2 weeks. Those findings were associated with reduction of glycogen contents in the myofibers of both muscles. 3. Widening of intermyofibrillar space, mitochondrial changes of electron-lucent matrix and indistinct cristae, and cisternal dilatation of sarcoplasmic reticulum were observed in both muscles after DMTU treatment. 4. Pretreatment of DMTU attenuated the ultrastructural changes induced by irradiation. Those were recoved normally by 2 weeks. Consequently, DMTU attenuates the ultrastructural changes in tibialis anterior and soleus muscle after irradiation. The more severe morphological changes in soleus muscle at 1 day and 3 days, and in tibialis anterior at 7 days and 2 weeks after irradiation are associated with the reduction of glycogen contents.

Effect of (R-)-N6-phenylisopropyladenosine (RPIA) Pretreatment on the alteration of Netural Cell dhesion Molecule / 대한간질학회지

BACKGROUND: Various neuronal and glial factors which participate in neural differentiation, including neural cell adhesion molecule (NCAM), are upregulated in pathogenesis of temporal lobe epilesy (TLE).This study aimed to investigate hte effect of (R-)-N6-phenylisopropyladenosine (RPIA), an adenosine A1 receptor agonist, on the morphological alteration of NCAM immunoreactivity (IR) in limbic system of Kainic acid (KA)-induced epileptic rats. METHODS: Experiment animals were divided into control group, KA treatment only (10 mg/kg. i.p.)group, and RPIA pretreatment (100 microgram/kg. i,p, 10 min prior to injection of KA) group. Animals were sacrificed at 24 hours and 1 week after KA treatment. Luxol fast blue-cresyl violet stain for histopathological observation, and NCAM immunohistochemistry to study alteration of NCAM IR in limbic system were performed. RESULTS: Neuronal loss in CA1 and CA3areas of hippocampus, piridorm cortex, basolateral amygdala nucleus and lateral dorsal thalamic nucleus were induced by KA unjection, and thoes were reduced by RPIA pretreatment. Inrease of NCAM-IR was observed in interneurons of all hippocampal areas. except CA2 area, pirform cortex and basolateral amygdala nucleus at 24 hours after KA injection. and increased NCAM-IR was observed in cell membrane and processes of neuroglia, dentate granule cells and pyramidal cells in CA1 area of hippocampus. and neurons in piriform cortex, amygdala and lateral dorsal thalamic nucleus 1 week after KA injection, but those changes were milder than those at 24 hours after KA injection. RPIA pretreatment significantly reduced KA-induced NCAM-IR in hippocampal CA3, CA1 area, piriform cortex, amtgdala and lateral dorsal thalamic nucleus. CONCLUSION: We suggest that decrease of NCAM immunoreactivity is associated with neuprotective effects of RPIA on limbic system against KA neurotoxiciy.