Effect of Nitric Oxide on Neuropeptide Y Immunoreactivity in Rat Spinal Cord on the Allodynia of Inflammation Induced by Freund's Complete Adjuvant / 대한해부학회지

Injection of Freund's complete adjuvant (FCA) into peripheral tissue induces inflammatory responses with accompanying pain behaviors. Injection of FCA produced a significant mechanical allodynia over time and nitric oxide(NO) is involved in this mechanism. The role of neuropeptide Y (NPY) on allodynia induced by inflammation is still controversal. We invastigated the change of spinal NPY and nitric oxide in rats with inflammation induced by subcutaneous injection of FCA and L-NG-nitro arginine methyl ester (L-NAME) into hind paw. The results are: The number of NADPH-diaphorase positive neurons and staining intensity of area increased at ipsilateral spinal ventral/dorsal horn of inflammation model. No significant changes were found with L-NAME posttreatment. Staining intensity of NPY immunoreactive (ir) area increased at ipsilateral spinal dorsal horn after FCA injection. No significant changes were found with L-NAME posttreatment. NPY-ir and NADPH-d reactive neurons were found in Rexed III-IV lamina at spinal dorsal horn. No significant change were found on all groups. The results suggest that spinal NPY is 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 NPY in spinal cord level.

Effect of Nitric Oxide on the Change of Spinal Neuropeptide in the Inflammation Model by Freund's Complete Adjuvant / 대한해부학회지

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.

The Effect of IGF-I on Collagen Synthesis in Cultured Chondrocytes

Cartilage is one of the most commonly manipulated tissue in esthetic and reconstructive surgery. Cartilage has an important role in longitudinal bone growth. Anabolic hormones and locally produced peptide growth factors are known to influence this process Matrix composition changes through proliferation, maturation, and differentiation of chondrocytes, and endochondral ossification thereafter. Defined cartilage matrix is synthesized during the maturation of chondrocytes where the major change is the increment of type II collagen. Variable sulfated mucololysaccharides and hyaluronic acid are also synthesized during this maturation. IGF-I(insulin like growth factor-I), so called somatomedin C, is a prominent growth factor in serum. IGF-I is known to be involved in long growth. IGF-I is affected by pituitary growth hormone. There are few studies done on IGF-I effect in cartilage matrix formation and possible changes of collagen subtypes. This experiment was designed to see the IGF-I effect on the colagen synthesis of cultured chondrocytes. Optimal concentration of IGF-I for the experiment was determined using H3-thymidine incorporation into DNA. The IGF-I effect on collagen synthesis was studied using H3-proline. The IGF-I effect on the synthesis of subtypes of collagen was studied using SDS-PAGE and immunocytochemical staining. Chondrocytes were isolated from the ears of New Zealand white rabbit and cultured in 2 X 10(5) cells/300 microgram density. IGF-I increased DNA synthesis, and optimal concentration of IGF-I was determined by dose-relationship curve as 10ng/ml. Collagen synthesis was increased by IGF-I. Type II collagen was increased on SDS-PAGE with IGF-I and this gel electrophoresis showed type X collagen, also. The increase in type II collagen was confirmed with immunocytochemical staining, the reaction becoming stronger with the addition of IGF-I. Type I collagen was not changed with IGF-I on immunocytochemistry. We conclude that IGE-I is an important modulator influencing not only proliferation and maturation but also terminal different-iation of chondrocytes.

Studies on the Paneth Cell of the Mouse Ileum after Administration of Cyclophosphamide / 대한해부학회지

This study was carried out on ICR mice, male, weighing about 26~35 g in order to investigate the effects of cyclophosphamide (CP) on the Paneth cells and their glycoconjugates. They were given intraperitoneally CP (Sigma, USA) 150 mg/kg body weight. Control mice were given as same amount of distilled water. The mice were sacrificed after 12 hours and on day 1, 2, 4, 6, 9 and 14 after CP injection. Sections were prepared from the region upper 1~2 cm from the end of the ilea. The material for histological examination was fixed in 10% buffered formalin. Some of the preparation 4 mm thick from the paraffin blocks stained with hematoxylin and eosin. The average numbers of the cells (Paneth cell index : PCI) were counted in the longitudinally sectioned 20 intestinal glands and semiquantitive granulation indices (Paneth cell granulation index : PGI) were obtained arithmatically weighted method to 3 cell types classified according to the degree of granularity of the cells. The other sections were incubated with 8 species of lectins (GS I B4, PSA, SBA, sWGA, UEA I, ECL, PNA and LFA). In order to increase the specificity of the reactions, the sections were applicated with ABC system. And then the sections were incubated DAB and were counterstained with hematoxylin. The results observed by light microcope were as follows. 1. The Paneth cell index (PCI) was 155.5 in control mice, while the PCI from the mice after 12 hours CP injection was 88.3. The Paneth cell granulation index (PGI) decreased from 316.0 in control mice to 152.3 in 12 hours after the CP administration. 2. The PCI increased to 141 and the PGI was 354 on day 2 after CP administration, which was higher in number than those of the control mice. It was characterized that the Paneth cells packed with numerous eosinophilic granules in the apical region increased in great numbers on day 2. 3. The PCI and PGI decreased on day 4 and day 5, and began to increase on day 9, which recovered to the similar level of the control mice. 4. Apoptotic-like cells increased suddenly in great numbers 12 hours after the CPA injection and began to decrease on day 1. Most of the dying cells seem to come from stem cells of the crypts and a small numbers of them from Paneth cells. 5. Paneth cells exhibited an extensive binding pattern for SBA, sWGA, and showed a restricted binding pattern for GS I B4 and UEA I. PSA, PNA, LFA. ECL showed negative reaction with Paneth cells. 6. It seems that Paneth cells can be classified according to the composition of the glycoconjugate in the granule and the stages of the cell maturation. The glycoconjugates in the halo is thought different from that in the core of the secretory granules. 7. The Paneth cell granules generally showed stronger reaction with the lectins in 12 hours after the CP admini-stration. 8. It is thought that the core of the granules decomposed earlier than the halo of the granules, and the granules of the cells reacted negatively with the lectins secreted earlier than those of the cells showed strong reaction with the lectins after the CP injection.

Morphological Effect of Chronic Alcohol Drinking upon the Gastric Mucosa of Rats / 대한해부학회지

In order to investigate the effect of chronic alcohol drinking upon the gastric mucosa of rats, Sprague-Dawley rats (200~235 g) had drunken 10% alcohol in stead of water and were maintained on regular feed. They drank about 20cc of 10% alcohol a day. The experimental groups sacrificed in 1, 2, 4 and 8 weeks. The gastric mucosa from the gastric body near by the fundus and the gastric body in the vicinity of the pylorus were fixed in neutral formalin, dehydrated in graded ethanol and embedded in paraffin. They were sectioned and stained with hematoxylin and eosin, and stained with alcian blue pH 2.5. The results were as follows: 1. Mucosal epithelium of the rat destroyed a little in some place, but the damage was not severe in all the experimental groups. Migrating cells and parietal cells covered the damaged places. 2. In the mucosa of the gastric body near by the fundus, the cells of the gastric gland showed positive reaction with alcian blue pH 2.5 and increased in number at 1 week. The number and reactivity of the cells decreased at 4 and 8 weeks of the experiment. 3. The surface mucous cells of the gastric pits in the body near by the pylorus decreased in number. The mucous neck cells of the gastric gland also decreased in number with time. 4. The cells in the base of the gastric gland showed alcian blue pH 2.5 positive at 4 and 8 weeks of the experiment. 5. The thickness of the gastric mucosa in all the experimental groups decreased in comparison with that of normal gastric mucosa.

Detection and Characterization of Enteroviral RNA in Paraffin-embedded Heart Tissues form Patients with Dilated Cardiomyopathy

The aim of this study was to investigate viral etiology in dilated cardiomyopathy (DCM) by polymerase chain reaction (PCR) or nested reverse tanscription PCR (RT-PCR), and characterize the enteroviral RNA presented in the clinical specimens. Twenty-eight paraffin-embedded heart tissue samples were assayed to detect cytomegalovirus, herpes simplex virus type 1, type 2, parvovirus, adenovirus, and enterovirus (EV) with each specific primer. Of these 28 patients (mean age: 27, M: 24, F: 4), 26 were histologically diagnosed as DCM and 2 as myocardial infarction (MI). Nested RT-PCR detected enteroviral RNA in 7 (26.9%) of 26 patients with DCM, and none of patients with MI. And none of DNA viruses tested were detected from the samples. Amplified products were also genotyped by single-variation of EV is present in the explanted heart tissues from patients with DCM. Although most of the sequences among the wild isolates have the greatest similarity to those of coxsackievirus B3, there are specific regions of variable sequences (no 490 - no 510). The data suggest that enterovirus may be a major viral pathogen for the DCM in Korea and nucleotide sequence data indicate that coxsackievirus B3 may be a leading etiologic agent of DCM.

Fine Structure and Glycoconjugate Complex Pattern of the Rat Vaginal Epithelium during Pregnancy / 대한해부학회지

The present study is designed to study the light and electron microscopic structure of the rat vaginal epithelium during pregnancy. Furthermore, lectin histochemistry is used to know the changes of the terminal sugar of the glycoconjugate in the vaginal epithelium during pregnancy. The 0 day of pregnacy is defined as the day of presence of sperm in a vaginal smear. At 0 day of pregnancy, most of cells are flat in morphology except basal cell. Lightly-stained superficial cells had few cell organelles, and all the other cells had many intermediate filament bundles, microvilli-like processes and desmosomes. During pregnancy, the thickness of the vaginal epithelium was increased. The morphology of the mucous cells are changed from a cuboidal shape to a columnar one. The intermediate filament bundles are decreased in the mucous cell after first week of pregnancy. Lectin histochemistry showed the presence of alpha-L-fucose, alpha-D-galactose, beta(1, 4)-N-acetylglucosamine, alpha-D-N-acetylgalactosamine and galactosyl-beta(1, 4)-N-acetylglucosamine in the mucous cells. The basal cells also contained the same terminal sugars except galactosyl-beta(1, 4)-N-acetylglucosamine. Approaching to the birthday, the thickening of the mucous layer of the vaginal epithelium suggests that the mucous containing several glycoconjugates may play an important role to make the appropriate environment in the vaginal lumen during pregnancy and parturition.

Effect of VCAM-1 expression in human endothelial cells by proinflammatory cytokines / 천식및알레르기

BACKGROUND: The expression of adhesion molecules contribute to development of systemic diseases. Vascular cell adhesion molecule-l(VCAM-1) is an endothelial cell membrane glycoprotein that has been implicated in leukocyte/endothelial cell interactions in inflammation. OBJECTIVE: The aim of this study was to characterize the surface expression and regulation of VCAM-1 on two different endothelial cells. METHOD: We examined the effects of the expression of VCAM-1 in two different endothelial cells, isolated from human umbilical cords and human glomerulus. Expression of VCAM-1 was measured by enzyme-linked immunosorbent assay(ELISA) and flow cytometry. RESULTS: In human umbilical cord endothelial cells(HUVECs), both interleukin-l B(IL-lB) and tumor necrosis factor-a (TNF-a) increased VCAM-1 expression. VCAM-1 expression increased by TNF-a was higher than that increased by IL-lB. In human glomerular endothelial cells(HGECs), IL-lB and TNF-a markedly increased VCAM-1 expression. Conclusion. The regulation of VCAM-1 appears to be somewhat different in HGECs compared with HUVECs. These differences between the responsiveness of the two cells may possibly indicate inherent differences in endothelial cell derived from different vascular beds.

Distribution of inositol 1,4,5-trisphosphate 3-kinase in the cat brain by immunohistochemical study / 대한해부학회지

No abstract available.