1.Experimental Study of Changes of beta-catenin, PCNA, Substance P after Peripheral Nerve Compression in Rat.
Young Churl YANG ; Seung Jae LEE ; Kwan Hee HONG
Korean Journal of Physical Anthropology 2001;14(1):97-115
The ultrastructural changes of sciatic nerve and immunohistochemical changes of beta-catenin, PCNA, substance P were studied at the proximal segment of rat sciatic nerve after compression injury. We used 90 Sprague Dawley rats and the sciatic nerve compressed using silicon tube. We divided experimental groups which were the compression group for 1 hour (1C), for 2 hours (2C), and for 3 hours (3C), the release group for 1 day (1C1R) and 3 days (1C3R) after the compression for 1 hour, the release group for 1 day (2C1R) and 3 days (2C3R) after the compression for 2 hours, the release group for 1 day (3C1R) and 3 days (3C3R) after the compression for 3 hours. The rats were sacrified and took the sciatic nerve specimen. The specimens were investigated under the light microscope after hematoxylin & eosin, toluidin blue, and immunohistochemical stainings. In the H & E finding, the axon of the 1C disappeared, but recovered at the 1C3R. The part of nerve fibers at the 2C were swollen, but began to be partially recovered at 2C3R. Most nerve fibers were enlarged at the 3C, but markedly decreased at the 3C1R. The beta-catenin reaction disappeared at the 1C, but almost recovered at the 1C3R. This reaction of the 2C disappeared in the large fibers, but began to be recovered in the small fibers at the 2C1R. This reaction of the 3C disappeared in the large fibers, but began to be recovered at the 3C1R and 3C3R. The PCNA reaction prominently appeared at the 1C3R and 2C3R, the more prominent reaction at the 3C1R, and markedly increased reaction at the 3C3R. The substance P reaction of the 1C1R was mild positive, and the 2C1R and 3C1R were strong positive. In the toluidin blue staining, the myelin sheaths near the perineurium began to be thickened at the 1C, but almost recovered at the 1C3R. Many myelin sheaths became to be very thickened at the 2C and 3C, but almost recovered at the 2C3R and 3C3R. In the electron microscopic findings, the myelin sheaths of the 1C underwent the demyelination with the separated lamellae and the increase microtubules. At the 1C3R, the axolemma was attached on the myelin sheath and the axon was recovered. the myelin sheaths of the 2C underwent the demyelination with the separated axolemma. At the 2C1R, the myelin sheath was recovered by the developing Schwann cells, many intraaxonal mitochondria of demyelinated nerve fibers. At the 2C3R, the myelin sheath tended to be recovered by the increased rough endoplasmic reticulum and mitochondria of Schwann cells, many intraaxonal mitochondria of demyelinated nerve fibers. The myelin sheaths of the 3C began to be underwent severe demyelination from the middle portion of the sheath and the vacuolization of intraaxonal mitochondria. At the 3C1R, the myelin sheaths were recovered and contained many extended microtubules, mitochondria, and small granules. At the 3C3R, severe demyelinated nerve fibers were recovered by increasing microtubules. The proximal retrograde degeneration of sciatic nerve by the acute compression appeared the loss of the axons and the swelling of nerve fibers. The beta-catenin reaction was disappeared by the compression, but recovered by releasing. This reaction may be played a important role of the recover of demyelination. The PCNA reaction of Schwann cells was increased by the nerve compression. In the substance P finding, the pain after the compression appeared at the 1 day after releasing. Electron microscopic changes after sciatic nerve compression were the demyelination, the separated lamellae and the increase of intraaxonal microtubules. After releasing, the nerve fibers were recovered by developing Schwann cell, the intraaxonal mitochondria, and the transported granules through extending microtubules.
Animals
;
Axons
;
beta Catenin*
;
Demyelinating Diseases
;
Endoplasmic Reticulum, Rough
;
Eosine Yellowish-(YS)
;
Hematoxylin
;
Microtubules
;
Mitochondria
;
Myelin Sheath
;
Nerve Fibers
;
Peripheral Nerves*
;
Proliferating Cell Nuclear Antigen*
;
Rats*
;
Rats, Sprague-Dawley
;
Retrograde Degeneration
;
Schwann Cells
;
Sciatic Nerve
;
Silicones
;
Substance P*
2.Effect of Inorganic Polyphosphate on Cultured Periodontal Ligament Cells.
Korean Journal of Physical Anthropology 2001;14(1):89-96
This study was designed to examine the effect of polyP on the periodontal ligament fibroblasts. The cells were incubated for 24 and 48 hours with various concentrations (0.001%, 0.002%, 0.005%, 0.010%, 0.020%, 0.040%) of polyP. Cell activity, alkaline phosphatase activity, and protein synthesis activity of the cells were examined. The results were as follows: 1. Twenty four hours after incubation with polyP, the cell activity of the periodontal ligament fibroblasts was increased. Forty eight hours after polyP application, the activity of the cells was similar to that of control group. 2. Twenty four or forty eight hours after incubation with polyP, the alkaline phosphatase activity of the periodontal ligament fibroblasts was increased. 3. The protein synthesis of the periodontal ligament fibroblasts was not changed after polyP application. These results suggest that, polyP may increase the cell activity and alkaline phosphatase activity of periodotal ligament fibroblasts and polyP may not affect the protein synthesis of the cells.
Alkaline Phosphatase
;
Fibroblasts
;
Ligaments
;
Periodontal Ligament*
;
Polyps
3.Seizure -Related Change of NADPH -diaphorase and Calcium Binding Protein Positive Neurons in the Brain of Rats.
Sun Young SHIN ; Young Buhm HUH ; Kyoung Lan KANG ; Chan PARK ; Min Jeong KANG ; Jae Ryong LEE ; Jung Hye KIM ; Jin Hwa YOO ; Hee Kyung AHN
Korean Journal of Physical Anthropology 2001;14(1):79-88
Nitric oxide (NO) is a gaseous messenger that plays a role in neurotransmission, long term potentiation, depression and cerebral blood flow. Increases in intracellular calcium levels activate the enzyme NOS, and the NO released then diffuse to adjacent cells and activate guanylate cyclase. NO mediates the increase in cerebral blood flow during seizure activity. Therefore, the present study was aimed to investigate the change of NOS and calcium binding proteins in the rat cerebral cortex following seizure. Rats were injected with kainate (KA) and killed at 6 hours, 1, 3, 5 and 10 days after seizure. Expressional change of nNOS, calbindin D28k and parvalbumin was assessed by histochemistry, immunohistochemistry and microdensitometry in the rat brain. The intensity of the NADPH -d staining in rat cortical neurons showed a marked susceptibility to KA administration. At 6 hours and 3 days after seizure, the optical density of the NADPH -d staining was increased relative to the signal in saline treated control rats. At 5 and 10 days after seizure, the optical density of NADPH -d staining was not significantly different in most cortical regions compared to controls. In the hippocampus, the optical density of NADPH -d staining was highest at 5 days after seizure. The optical densities of calbindin D28k and parvalbumin positive neurons were various in the cerebral cortex, hippocampus and caudatoputamen during postseizure period. These results indicate that the calcium binding proteins investigated here are not essential for determining the activation of nNOS/NADPH -d positive neurons in the cerebral cortex and striatum.
Animals
;
Brain*
;
Calbindin 1
;
Calbindins
;
Calcium*
;
Calcium-Binding Proteins
;
Carrier Proteins*
;
Cerebral Cortex
;
Depression
;
Guanylate Cyclase
;
Hippocampus
;
Immunohistochemistry
;
Kainic Acid
;
Long-Term Potentiation
;
NADP*
;
Neurons*
;
Nitric Oxide
;
Rats*
;
Seizures*
;
Synaptic Transmission
4.A Study on the Early Development of the Hip Joint in Staged Human Embryos and Fetuses in Korean.
Kyun Ho CHANG ; Hyoung Woo PARK ; Won Kyu KIM ; Doo Jin PAIK
Korean Journal of Physical Anthropology 2001;14(1):61-78
The author has studied 33 cases of Korean embryos of Carnegie stage 11 ~23 and 18 cases of fetuses to demonstrate the development of the hip joint. The external feature of the lower extremity was observed by stereoscope and digital camera, and the internal structures were studied by light microscopic observation. The results obtained were as follows: In stage 13 lowerlimb buds were appeared. In stage 17 mesenchymal condensation for femur and hip bone, and one -layered interzone were observed. In stage 18 cartilage models for ilium and ischium were visible. In stage 22 three -layered interzone between the head of femur and hip bone was formed. In stage 23 acetabular labrum and distinct three -layered interzone was visible. In the 9th weeks mesenchymal ligamentum capitis femoris and transverse acetabular ligament are appeared, and acetabular labrum was reacted tracely to trichrome stain. In the 10th week the joint space was formed between the femoral head and hip bone, and shallow depression in acetabulum is found. In the 12th week the articular capsule was directed lateral to acetabular labrum, and numerous blood vessels were found in acetabular fossa and ligamentum capitis femoris, and cartilage canal were developed in femoral neck. In the 14th week cartilage canal was found in middle of femoral head, and synovial fold were developed, and ligamentum capitis femoris was shown strongly positive reaction. In the 16th week cartilage canals were more found, and numerous blood vessels were observed in fovea capitis. In the 18nd week the neck of femur was narrow, and femoral head was lied deeply in acetabulum with acetabular labrum. Consequently the lower extremity of Korean embryos and fetuses was first appeared in stage 13, and hip joint development was started at stage 17. The articular cavity was first formed at the 9th week of development, the acetabulum labrum was developed at stage 23. The mesenchymal ligamentum capitis femoris was appeared at the 9th week. At the same time the transverse acetabular ligament fully encircled the femoral head. The articular capsule has lined the articular cavity at the 12th week, and synovium was formed at the 14th week of development. At the 18th week the hip joint has attained its final shape.
Acetabulum
;
Blood Vessels
;
Cartilage
;
Depression
;
Embryonic Structures*
;
Femur
;
Femur Neck
;
Fetus*
;
Head
;
Hip Joint*
;
Hip*
;
Humans*
;
Ilium
;
Ischium
;
Joint Capsule
;
Joints
;
Ligaments
;
Lower Extremity
;
Neck
;
Synovial Membrane
5.Protective Roles of Adenosine A1 Receptor Agonist and KATP Channel Opener in Adriamycin induced Cardiac Toxicity.
Sahng LEE ; Wan Gil CHO ; Doo Jin PAIK ; Bang Hun LEE ; Tai Kyoung BAIK ; Won Kyu KIM ; Ho Sam CHUNG
Korean Journal of Physical Anthropology 2001;14(1):45-59
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.
Adenosine*
;
Animals
;
Antioxidants
;
Apoptosis
;
Creatine Kinase
;
Doxorubicin*
;
Glyburide
;
Heart
;
Heart Failure
;
Immunohistochemistry
;
In Situ Nick-End Labeling
;
Mice
;
Mice, Inbred ICR
;
Muscle Cells
;
Myocardium
;
Myocytes, Cardiac
;
Oxidative Stress
;
Pinacidil
;
Reactive Oxygen Species
;
Receptor, Adenosine A1*
;
Thiourea
6.Localization of Laminin and Laminin beta 1 chain on Gomeruli of Developmental Rat Kidney.
Ho Sam CHUNG ; Youn Kyung SEO ; Doo Jin PAIK ; Won Kyu KIM ; Jee Hee YOUN ; Ho Jung KIM ; Tai Kyoung BAIK ; Won Jae ROH
Korean Journal of Physical Anthropology 2001;14(1):29-44
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.
Animals
;
Basement Membrane
;
Capillaries
;
Collagen Type IV
;
Cytoskeleton
;
Doxorubicin
;
Endoplasmic Reticulum, Rough
;
Endothelial Cells
;
Epithelium
;
Extracellular Matrix
;
Fetus
;
Glomerular Basement Membrane
;
Glomerulonephritis
;
Glycoproteins
;
Heparan Sulfate Proteoglycans
;
Humans
;
Infant
;
Infant, Newborn
;
Kidney*
;
Laminin*
;
Mesangial Cells
;
Microscopy, Electron
;
Nephrons
;
Nuclear Envelope
;
Parturition
;
Rats*
;
Regeneration
7.Effects of c -Fos Immunoreactivity on Central Nervous System after Administration of Aspirin.
Chang Mok SON ; Chul Hong KIM ; Ki Soo YOO
Korean Journal of Physical Anthropology 2001;14(1):17-27
Aspirin is one of the popular non -steroid anti -inflammatory drugs used in the management of pain. This study was performed to investigate the effects of aspirin on c -Fos expression in rat CNS after inducing somatic pain with formalin. Male S.D. rats were injected subcutaneously with 0.1 ml of 5% formalin in the plantar surface of right hindpaw. For experimental group, aspirin was administered orally before injection of formalin. Asprin -untreated group was utilized as the control group. Rats were sacrificed at 0.5, 1, 2, 6 and 24 hours after formalin injection. Rat brains were removed and sliced in rat brain matrix. Brain slices were coronally sectioned at interaural 5.70 ~6.70 mm. Serial sections were immunohisto-chemically reacted with polyclonal c -Fos antibody. The numbers of c -Fos protein immunoreactive neurons in the cingulate cortex, primary somatosensory area, and hippocampus were counted and analyzed statistically with Mann - Whitney U test. Results were as follows: 1. Higher numbers of c -Fos immunoreactive neurons were found in the cingulate cortex, primary somatosensory area and hippocampus. 2. Both aspirin -treated and -untreated groups, numbers of c -Fos immunoreactive neurons were significantly higher all time points than formalin -untreated group, which peacked at 2 hours. 3. The numbers of c -Fos immunoreactive neuron of the aspirin -treated group were less compared to the aspirin - untreated group at each time point. In conclusion, these results provide some basic knowledge in understanding the mechanism and control of formalin - induced somatic pain.
Animals
;
Aspirin*
;
Brain
;
Central Nervous System*
;
Formaldehyde
;
Gyrus Cinguli
;
Hippocampus
;
Humans
;
Male
;
Neurons
;
Nociceptive Pain
;
Rats
8.Effects of Maternal Hyperthermia on the Palatal Mesenchyme Development of Hsp70 Knock-out Mice Fetuses.
Korean Journal of Physical Anthropology 2005;18(2):115-130
To investigate the effects of maternal hyperthermia on the development of the palate, pregnant Hsp70 knock-out mice at gestational day (GD) 8.5 were immersed in 43degrees C water bath until their body core temperature reached at 43degrees C. Thereafter, pregnant mice were given more 5 minutes hyperthermic exposure. Heat-untreated Hsp70 WT mice fetuses were used as the control group. Fetuses were collected at embryonic day 13.5, 14.5 and 15.5 (E13.5, E14, 5 and E15.5). Heads followed by removal of the mandible and the tongue were obtained and photographed for palatal development. Developing palates were processed for histological and immunohistochemical studies. Tissue sections were immunostained for TGF-beta2, FGF-8 and fibronectin, and observed with light microscope. The obtained results were as follows: Cleft palate was formed in heat-treated Hsp70 KO fetuses at E14.5 and E15.5. Immunohistochemical findings indicated that TGF-beta2 expression of the experimental fetuses were more delayed than that of the control fetuses. Mesenchyme under the medial edge epithelium (MEE) and cells of MEE showed continuously strong positive TGF-beta2 reactivity at E15.5. FGF-8 was revealed in both of the mesenchyme and the epithelium at the same time. FGF-8 immunoreactivity in the mesenchyme and the epithelium of the heat-treated fetuses showed strong reactivity at E15.5. In the experimental fetuses fibronectin was revealed the mesenchyma and basal lamina at E15.5. Taken together, it is suggested that maternal hyperthermia induces continuous expression of TGF-beta2 and FGF-8 in the mesenchyme and delayed expression of fibronectin. These should affect the normal palatogenesis and result in cleft palate.
Animals
;
Basement Membrane
;
Baths
;
Cleft Palate
;
Epithelium
;
Fetus*
;
Fever*
;
Fibronectins
;
Head
;
Immunohistochemistry
;
Mandible
;
Mesoderm*
;
Mice
;
Mice, Knockout*
;
Palate
;
Tongue
;
Transforming Growth Factor beta2
9.Effects of Maternal Hyperthermia on the Development of C57/BL6 Strain Hsp70 Knock-out Mice Fetuses.
Korean Journal of Physical Anthropology 2005;18(2):105-113
To investigate the effects of maternal hyperthermia on the formation of congenital anomalies, pregnant Hsp70 knock-out and wild type mice at gestational day (GD) 8.5 were immersed in 43degrees C water bath until their body core temperature reached at 43degrees C. Thereafter, pregnant mice were given more 5 minutes hyperthermic exposure. Pregnant mice were killed at GD 15.5, and fetuses were photographed for external appearance analysis. Fetuses with congenital anomalies such as anophthalmia and exencephaly were 72.6% (53 out of 73) in KO group and 28.2% (26 out of 90) in WT group, respectively. Histological findings showed exencephaly, eye abnormalities such as eyeball with retina only or buried eyeball or absence of eye structure, numerous apoptotic cells in the retina and inner ear neuroepithelium, cleft palate, and delayed endochondral ossification. The results of this study suggest that Hsp70 may have a protective function against heat shock.
Animals
;
Anophthalmos
;
Baths
;
Cleft Palate
;
Ear, Inner
;
Eye Abnormalities
;
Fetus*
;
Fever*
;
Hot Temperature
;
Mice
;
Mice, Knockout*
;
Neural Tube Defects
;
Retina
;
Shock
10.Effect of Exercise to Improve of Rat Lower Limb Healing after Physical Injury.
Duk Joon SUH ; Dae Hee LEE ; Hyun Hee SON ; Ji Young KIM ; Ki Soo YOO
Korean Journal of Physical Anthropology 2005;18(2):95-104
The aim of this report is to show that treadmill running exercise under well-controlled conditions is to improve of regeneration in rat gastrocnemius muscles after physical injury. For this, rats were submitted to bouts of exercise on a treadmill up a 10 degrees decline for 60 min and gastrocnemius muscles were analysed at different exercise periods by immunohistochemistry in comparison with injured nonexercised muscles. Rats were used with guidelines for experimental procedures as set forth in the Declaration of Helsinki. We analysed the regenerative processes by detection of immunoreactivity for the two intermediate filaments, desmin and vimentin. Desmin and vimentin are specific components of the cytoskeleton of striated muscle fibers and of mononuclear cells of mesenchymal origin including myoblasts, respectively. We found that non-exercised rats had more desmin-and vimentin-positive myofibers than that of exercised rats at 9th, 16th, 23th, 30th day after physical injury. At 30th day, non-exercised rats had several desmin-and vimentinpositive myofibers, but exercised rats had numerous normal myofibers. These results show that exercise is able to improve regeneration processes in physical injured gastrocnemius muscles of rats.
Animals
;
Cytoskeleton
;
Desmin
;
Helsinki Declaration
;
Immunohistochemistry
;
Intermediate Filaments
;
Lower Extremity*
;
Muscle, Skeletal
;
Muscle, Striated
;
Muscles
;
Myoblasts
;
Rats*
;
Regeneration
;
Running
;
Vimentin