• Applied Microscopy
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• v.24 no.2
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• pp.78-92
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• 1994

Lysozyme has been reported to be present in the secretory granules of the Paneth cell, and lysozyme immunoreactivity has been detected by immunogold method in Paneth cells of the intestine of human, mouse and rat. The present study was aimed at clarifying the intracellular distribution and changes of the lysozyme immunoreactivity in rabbit Paneth cell after common bile duct ligation of rabbit, using the electron microscope immunogold technique. Healthy adult rabbits weighing about 2kg body weight were divided into normal and bile duct ligated groups. Common bile duct ligation was performed aseptically under ether anesthesia. Experimental animals were sacrificed on the 1st, the 3rd, the 5th, the 7th and the 14th day after the operation. Mucosal specimens from the intestinal gland of ileum were fixed in 2.5% glutaraldehyde-1.5% paraformaldehyde, followed by 1% osmium tetroxide, embedded in araldite mixture, cut with LKB-V ultratome. Ultrathin sections were placed on parlodion coated nickel grids (200mesh). The section-bearing grids were floated upside down on the added substance in a moist chamber at room temperature except for the primary antibody step, which was at $4^{\circ}C$. Sections were etched with a saturated solution of sodium m-periodate for 60min. After etching, sections were pretreated with 0.02M tris buffered saline (TBS), pH 8.4, with 1% bovine serum albumin (BSA, Sigma) for 60min, then treated polyclonal rabbit anti-human lysozyme (Dakipatts) diluted 1 : 50 in TBS with 0.1% BSA for 20hr. Subsequently, grids were incubated 60min in biotinylated goat anti rabbit IgG (Amersham) diluted 1 : 100 in TBS with 0.1% BSA. After this, sections were incubated 60min on streptavidin gold G10 (Amersham) diluted 1 : 50 in TBS with 0.1% BSA. After each step, the grids were briefly rinsed with TBS with 0.1% BSA. After the strepavidin gold step, the sections were jet washed with distilled water. Counterstain of the sections performed by uranyl acetate and lead citrate, and observed with JEM 100 CX II electron microscope. Observed results were as follow; 1. Secretory granules of mouse Paneth cells have a lysozyme immunoreactivity and also eosinophil leucocyte of rabbit applied for the positive-control stain, are well labeld with gold particles. 2. Normal rabbit Paneth cells have a lysozyme immunoreactivity restricted on the secretory granules. 3. Amount lysosomes containing myelin figures in the Paneth cells were significantly increased from 5th day after the common bile duct ligation. 4. Immunoreactivity of Paneth cell secretory granules were more activated on the 3rd day after the common bile duct ligation as compared with those of the normal animal. But the lysozyme immunoreactivity were decreased from the 5th day after the common bile duct ligation. 5. Considering the above finding, lysozyme contained Paneth cell are affected following of common bile duct ligation, whereas lysosomes containing myelin-figure do not exhibit any immunoreactive relationship with those of secretory granules.

• The Korean Journal of Nuclear Medicine
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• v.32 no.3
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• pp.298-304
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• 1998

Purpose: We evaluated the usefulness of Re-188 sulfur colloid for radiation synovectomy and therapy of intraperitoneal metastasis. Materials and Methods: We investigated the labeling efficiency of Re-188 sulfur colloid on various conditions. The stability of Re-188 sulfur colloid was observed at room temperature for 24 h and in human serum and synovial fluid for 72 h. The particle size distribution of Re-188 sulfur colloid was measured by filtering with various pore size filters. Animal experiment was performed in mice and rabbits. Results: The labeling efficiency of Re-188 sulfur colloid was $64.5{\pm}5.8%$ (n=5) at the conditions of sodium thiosulfate 40 mg, EDTA $Na_2.2H_2O$ 0.8 mg, $KReO_4$ 0.8 mg at pH 1. After purification, the radiochemical purity was higher than 99%. The stability of Re-188 sulfur colloid was high (>99%) at room temperature for 24 h and in human serum and synovial fluid for 72 h. The particle size distribution of Re-188 sulfur colloid was 0.3% ($<1{\mu}m$), 11.2% ($1{\sim}5{\mu}m$), 25.8% ($5{\sim}10{\mu}m$) and 52.8% ($>10{\mu}m$). In mice, 1 h postinjection of Re-188 sulfur colloid into tail vein, uptakes in lung, liver and muscle were $37.30{\pm}5.36$, $32.33{\pm}1.79$, $6.60{\pm}0.02%$ ID/organ respectively. After i.p. injection in mice, the uptakes of extraperitonial organs of Re-188 sulfur colloid at 1 and 24 h were $0.1{\pm}0.1$, $0.4{\pm}0.1%$ ID/organ, and the excretions through urine and feces (${\sim}70 h$) were low ($2.68{\pm}0.80$, $0.95{\pm}0.17%$). When Re-188 sulfur colloid was injected to synovial space of rabbit, the uptake in other organs except knee was very low. Conclusion: Re-188 sulfur colloid showed high labeling efficiency, stability and potency for clinical use.

• Applied Biological Chemistry
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• v.8
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• pp.101-111
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• 1967

Daily doses of phenylmercuric acetate arranged in $30{\gamma}\;(group\;I)$, 3{\gamma}\;(group\;II)$ and $0.3{\gamma}\;(group\;III)$ were administered respectively to rabbits for 90 days. The chief histopathological changes in the organs and the analytical data on mercury residues in the excretion and liver were as follows. 1. Kidney: In group I, severe degrees of vacuolization and cloudy swelling were occurred in the epithelial cells of proximal convoluted tubules and severe cloudy swelling and coagulative necrosis were observed in the proximal straight tubules. There were many hyaline casts in the collecting tubules. In group II, moderate degrees of vacuolization and cloudy swelling were observed in the epithelial cells of proximal convoluted tubules and moderate cloudy swelling and coagulative necrosis were encountered in the proximal straight tubules. A little numbers of hyaline casts were located in the lumen of collecting tubules. In group III, slight degree of cloudy swelling were observed in the epithelial cells of proximal convoluted and straight tubules. 2. Liver: In group I, cloudy swelling, fatty changes and coagulative necrosis were observed in the central zone of hepatic lobules. Dissociation of hepatic cell cords was encountered. Hyperplsia of hepatic cells were remarkable in group II. No Pathological changes were observed in group III. 3. Spleen: Deposition of hemosiderin pigment was prominant in group I and small amount of the pigment was observed in group II. There were no pathological changes in group III. 4. Adrenal, colon and heart: No pathological changes were detected in all 3 groups. 5. In an average about 76.5% of mercury was excreted from group I, 85.4% from group II and 79.8% from group III. 6. Mercury content in the liver was 0.0348 g in group I, 0.00378 g and 0.00066 g in group II and group III respectively. 7. In general, as to increased mercury doses the concentration of mercury accumulation in the liver became higher, how·ever, the accumulation quantity against a total amount of mercury doses showed an adverse trend. In other word, the quantity of mercury accumulation was not increased proportionately by higher dose of mercury.

• Korean Journal of Microbiology
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• v.44 no.3
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• pp.178-185
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• 2008

Avian influenza virus (AIV) is recognized as key to the emergence of pandemic influenza for humans; there are growing concerns that AIV H9N2 may become more efficient to transmit to humans in the near future, since the infection of poultry with AIV H9N2 has been common in recent years. In this study, we aimed to produce antisera recognizing the HA and NA proteins of AIV H9N2. Initially, coding sequences corresponding to the N-terminal regions of the HA and NA proteins of the Korean AIV H9N2 (A/Ck/Kr/MS96/96) isolated from a domestic chicken were amplified from the genomic RNA. Following cloning of the amplified cDNA fragments into pGEX4T-1 vector, two GST-fusion proteins (GST-HAln and GST-NAn) were expressed in E. coli BL21 and purified with glutathione sepharose columns; the recombinant GST-HAln and GST-NAn proteins were both used as immunogens in rabbits. The antigenicity of the rabbit antisera was analyzed by immunoblotting of the cell lysates prepared from AIV H9N2-infected MDCK cells. Overall, the recombinant HAln and NAn proteins fused to the C-terminus of GST and the rabbit antisera raised against the corresponding recombinant proteins would provide a valuable reagent for AIV diagnosis and basic research.

• Tuberculosis and Respiratory Diseases
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• v.48 no.4
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• pp.500-512
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• 2000

Background : To evaluate the efficacy of two methods of obtaining lung recruitment to reduce ventilator-induced lung injury(VILI). Methods : Fifteen New-Zealand white rabbits were ventilated in the pressure-controlled mode while maintaining constant tidal volume(10 ml/kg) and fixed respiration rate. Lung injury was induced by repeated saline lavage (PaO2<100 mmHg), and the pressure-volume curve was drawn to obtain Pflex. The animals were then randomly assigned to three groups and ventilated for 4 hours. In the control group(n=5), positive end-expiratory pressure(PEEP) less than that of Pflex by 3 mmHg was applied throughout the study. In the recruitment maneuver(RM) group(n=5), RM(CPAP of 22.5 mmHg, for 45 seconds) was performed every 15 minutes in addition to PEEP level less than Pflex by 3 mmHg This phrase is unclear. In the Pflex group, PEEP of Pflex was given without RM. Gas exchange, lung mechanics, and hemodynamics parameters as well as pathology were examined. Results : 1) Both the control and RM groups showed decreasing tendency in PaO2 with time. There was significantly decreased PaO2 at 4 hr compared to Ihr(p<0.05). But in the Pflex group, PaO2 did not decrease with time(p<0.05 vs other groups at 3, 4 hr). PaCO2 did not show significant difference among the three groups. 2) There was no significant difference in static compliance and plateau pressure. Mean blood pressure and heart rate also did not show any significant difference among the three groups. 3) The pathologic exam showed significantly less neutrophil infiltration in the Pflex group than in the control group(p<0.05). There was borderline significant difference in hyaline membrane score among the groups (p= 0.0532). Conclusion : Although recruitment maneuver of the injured lung may be important in decreasing VILI, it alone may not be sufficient to minimize VILI.

• Journal of Life Science
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• v.28 no.10
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• pp.1220-1232
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• 2018

The purpose of this animal study was to evaluate, by histological analysis, bone regeneration in rabbit maxillary sinuses with an anorganic bovine graft (Bio-Oss) and a ${\beta}-tricalcium$ phosphate (${\beta}-TCP$) grafting. Bilateral sinus augmentation procedures were performed in 12 adult male rabbits. Rectangular replaceable bony windows were made with a piezoelectric thin saw insert. In the Bio-Oss group, Bio-Oss was grafted and in the ${\beta}-TCP$ group, ${\beta}-TCP$ was grafted and covered by replaceable bony windows. The animals were sacrificed at 2, 4, and 8 weeks after the surgical procedure. The augmented sinuses were evaluated by histomorphometric analysis using hematoxylin-eosin, Masson trichrome, and tartrate-resistant acid phosphatase stains and also by immunohistochemical analysis of proliferating cell nuclear antigen (PCNA), type I collagen, and osteocalcin content. Histologically, new bone formation was found on the surface of Bio-Oss and ${\beta}-TCP$ particles from 2 weeks and continued to 8 weeks. Significant higher new bone formation was revealed in the ${\beta}-TCP$ group than in the Bio-Oss group at 8 weeks. The amount of graft materials was significantly decreased in the ${\beta}-TCP$ group and the number of osteoclasts was significantly increased in the ${\beta}-TCP$ group from 4 to 8 weeks. Immunoreactivity to PCNA was reduced at 8 weeks. The expression of type I collagen was significantly increased in the ${\beta}-TCP$ group at 2 weeks, but was significantly increased in the Bio-Oss group at 8 weeks. Immunoreactivity to osteocalcin was increased from 2 to 8 weeks. These histological results can help in the selection of graft materials for implants. Both Bio-Oss and ${\beta}-TCP$ are proven graft materials, however, these results indicate that ${\beta}-TCP$ showed better bone regeneration results in rabbit maxillary sinus augmentation.

• The Korean Journal of Physiology
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• v.7 no.1
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• pp.23-31
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• 1973

Numerous factors concern with the absorption of substances through the membrane of the gastrointestinal tract. To simplify the experimental condition, present work has been restricted to observe the disappearance rate of substance from the intestinal loop which was made in the jejunum, 70 cm apart from the pylorus of the adult rabbit. The purpose of the study is to clarify the absorption of urea through the jejunal wall is solely attributable to the concentration difference between the luminal fluid and plasma, and to observe the effect of adding red pepper upon the rate of absorption. The rabbits were anesthetized with nembutal, 35mg/kg I.V. Jejunal loop was made by ligating at 2 spots, 70 cm and 80cm apart from the pylorus. After rinsing with normal saline solution through the polyethylene tubing inserted from the end of the loop, 8 ml of test solution was placed through the same tubing. The test solution contained 200 mg% of urea and 150mg% of polyethylene glycol(M.W. 4,000) in normal saline solution. Right after placing the test solution the first specimen was taken through the tubing, and successive samplings were performed at 5, 10, 20, and 30 minutes. Logarithm of the difference of urea concentration between the luminal fluid and plasma was plotted against time elapsed after the onset of the experiment. If straight line is revealed, it would verify the nature of transport mechanism as diffusion, obeying the Fick's principle. The concentration of polyethylene glycol (PEG) was also measured in order to examine the change in the volume. PEG was used as the marker substance because it is not absorbable in the intestinal tract. Consequently the concentration of PEG relates inversely to the volume of the loop. Instantaneous concentration of urea in the loop times the volume will give the amount of urea remaining in the luminal fluid. The change in the amount of any substance is directly relate to the volume of the compartment and differs from the change in the concentration which is independent of the volume. After completion of the experiment without red pepper, it was added in the test solution and was centrifuged after thorough mixing. Supernatant of the mixture was placed in the loop and similar sampling were performed with the same time intervals that of previous run in order to observe the effects of the red pepper on the passive transport of the water soluble small substance, urea. The results obtained were as follows: 1. Logarithm of the concentration difference of urea between the luminal fluid and plasma was diminished exponentially as time elapsed. The decay constant in the experiment without red pepper was 0.0563/min. By adding red pepper in the test solution as much as the concentration rose to 4,000 mg% and 8,000 mg%, the decay constants were lowered to 0.0493/min and to 0.0506/min, respectively. The time interval by which the concentration difference dropped to one half of the initial value was prolonged. Without red pepper the half concentration time was 13.30 minutes, and by adding extract of red pepper, 15.31 minutes and 15.71 minutes were revealed. 2. The profile of the diminishing rate of tile amount of urea was quite different from that of the concentration because of the change in the volume of the loop during the observed period. 3. By adding the extract of red pepper, it slowed down the rate of absorption of urea in the intestinal loop, suggesting an increase in the diffusional barrier. 4. Larger dosage of red pepper brought an increase in the secretion of intestinal fluid with concomitant expansion of the luminal volume, and the retardation of the absorption of urea was noticed. This effect was largely dependent on the sensitivity of the individual animal to the red pepper, extract. The amount of urea remained after 10 minutes interval was 55.5% of the initial amount in the experiment without red pepper. On the other hand it was not consistent after administration of red pepper, showing 50.6% and 66.5% of the initial figures by adding 400 mg and 800 mg of red pepper in the test solution, respectively. It was postulated that symptom of diarrhea often encountered by taking a hot (red pepper) food might be attributable to the increase of secretion and the retardation of absorption in the intestinal tract.

• The Journal of Korean Academy of Prosthodontics
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• v.47 no.4
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• pp.445-456
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• 2009

Statement of problem: Many studies have been conducted to improve the primary stability of implants by providing bioactive surfaces via surface treatments. Increase of surface roughness may increase osteoblast activity and promote stronger bonding between bone and implant surface and it has been reported that bioactive surface or titanium can be obtained through alkali and heat treatment. Purpose: The purpose of this study was to evaluate the stability of alkali and heat treated implants via histomorphometric analysis. Material and methods: Specimens were divided into three groups; group 1 was the control group with machined surface, the other groups were treated for 24 hours in 5 M NaOH solution and heat treated for 1 hour at $600^{\circ}C$ in the atmosphere (group 2) and vacuum (group 3) conditions respectively. Surface characteristics were analyzed and fixtures were implanted into rabbits. The specimens were histologically and histomorphometrically compared according to healing periods and change in bone composition were analyzed with EPMA (Electron Probe Micro Analyzer). Results: 1. Groups treated with alkali and heat showed increase of oxidization layer and Na ions. Groups 2 which was heat treated in atmosphere showed significant increase of surface roughness (P<.05). 2. Histomorphometric analysis showed significant increase in BIC (bone to implant contact) according to increase in healing period and there was significant increases in groups 2 and 3 (P<.05). 3. BA(bone area) ratio showed similar results as contact ratio, but according to statistical analysis there was significant increase according to increase in healing period in group 2 only (P<.05). 4. EPMA analysis revealed no difference in gradation of bone composition of K, P, Ca, Ti in surrounding bone of implants according to healing periods but groups 2 and 3 showed increase of Ca and P in the initial stages. Conclusion: From the results above, it can be considered that alkali and heat treated implants in the atmosphere have advantages in osseointegration in early stages and may decrease the time interval between implantation and functional adaptation.

• Investigative Magnetic Resonance Imaging
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• v.13 no.1
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• pp.81-87
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• 2009

Purpose : We sought to evaluate enhancement of plaque with gadolinium-based contrast agent by magnetic resonance imaging (MRI) in comparison with histopathology, namely lipid-rich and macrophage-rich components that were two representative characteristics of plaque vulnerability using atherosclerotic rabbit aorta in order to determine which histopathologic component is relevant to the enhancement. Materials and Methods : New Zealand white rabbit (n=4, weight 3.0 to 3.5 kg, all male) was used for animal model of atherosclerosis. Atherosclerotic aortic lesions were induced by high-cholesterol diet and double balloon injury. T1-weight axial images were acquired before and after gadolinium-based contrast agent using a 3-T MRI. MR images and the matched histopathological sections (n=35) were divided into 4 quadrants or 3 (n=130). Enhancement ratio (ER, ER=SIpost/SIpre) on MRI was calculated for each quadrant and compared with histopathology in regard to lipid-rich and macrophage-rich areas. Results : Lipid-rich quadrants were 72 and fibrous quadrants were 58. The number of quadrants which had macrophage-rich areas was 105 and that of quadrants which did not have macrophage-rich areas was 25. ER was significantly higher in lipid-rich quadrants than in fibrous quadrants (mean ER 2.25c$\pm$0.41 vs. 2.72$\pm$0.65, p=0.013). ER poorly correlated with macrophage-rich areas when lipid-component was controlled (correlation coefficient -0.203, p=0.236). Conclusion : Lipid-rich plaques showed stronger enhancement than fibrous plaques using a standard gadolinium-based extracellular contrast agent. Macrophage infiltration did not correlate with degree of enhancement. Further study is warranted that account for optimal time of imaging after contrast injection using various plaque models from early to advanced stages and all possible parameters associated with contrast enhancement.

• Tuberculosis and Respiratory Diseases
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• v.49 no.6
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• pp.703-714
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• 2000

Background : Surfactant protein A (SP-A) is important in the regulation of surfactant secretion, synthesis and recycling. SP-A has important roles in regulating surfactant metabolism as well as in determining surfactant's physical properties. Since systemic sepsis is one of the common causes of acute respiratory distress syndrome (ARDS) and abnormalities in surfactant function have been described in ARDS, the authors investigated the effects of endotoxemia on the accumulation of mRNA encoding SP-A and SP-A protein content. Methods : Adult rats were given various doses of intraperitoneal endotoxin from Salmonella enteritidis and sacrificed at different times. SP-A mRNA was measured by filter hybridization method. Lung SP-A protein content was determined by double sandwich ELISA assay using a polyclonal antiserum raised in rabbits against purified rat SP-A. Results : 1) The accumulation of SP-A mRNA in the endotoxin treated group 24 hours after 2mg/kg and 5mg/kg endotoxin treatments was significantly increased 50.9% and 27.3%, respectively, compared to the control group (P<0.001, P<0.025). 2) The accumulation of SP-A mRNA 24 hours in the 5mg/kg endotoxin treated group was significantly increased by 26.5% compared to the control group (P<0.01). 3) Total amount of lung SP-A was not altered at 24 hours by various doses of treatment. Total lung SP-A content 144 hours after endotoxin administration was significantly decreased by 51.4% compared to the control group (P<0.01). Conclusions : The specific regulation of SP-A by various time course in vivo is evident. The late decline in SP-A protein content was unexpected and suggests that SP-A may be differentially regulated during lung inflammation. The functional significance of these alterations remains to be clarified.