• Journal of Chest Surgery
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• v.30 no.6
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• pp.559-565
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• 1997

Donor airway ischemia is a significant problem after tracheal replacement with homograft or lung transplantation, Several factors such as omentopexy, heparin, PGl2 and fibroblast growth factor, have been shown to induce angiogenesis in vitro and in vivo. This study was designed to investigate whether omentopexy and basic flbroblast growth factor can enhance rabbit tracheal revascularization and epithelial regeneration, Three different experiments were performed with New Zealand white rabbit. In group I(n= 15 control group), only coNical tracheal autotransplantation was done. In group II(n= 15), cervical tracheal autotransplantation with omentopexy was done through subcutaneous route. In group III(n= 15), cervical tracheal autotransplantation was done and lug basic flbroblast growth factor was applied. After 3, 7 and 14 days, the animals were sacrificed. The extent of revascularization was investigated by means of uptake of the human serum albumin labelled with 99m technetium, and epithelial regeneration were assessed by means of light microscope. In the group investigated at day 3, there was statistically significant high tracheal revascularization in group III(p<0.05), but no difference at 7 and 14 days. And epithelial regenerations at day 3 were better in group III(p<0.05), and at day 7 in group II and III. But there was no difference at day 14. We concluded that b-FGF can enhance the revascularization and epithelial regeneration of the tracheal autograft especially in early phase.

• Journal of the Korean Society of Food Science and Nutrition
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• v.31 no.6
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• pp.1084-1091
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• 2002

Mortierella alpina, a common soil fungus, is the most efficient organism for production of production acid presently known. Since arachidonic acid are important in human brain and retina development, it was undertaken the growing effect containing diet as a food ingredient. Arachidonic acid rich oil derived from Mortierella alpina, was subjected to a program of studies to establish for use in diet supplement. This study was compared the growth and learning effect of fungal oil rich in arachidonic acid by incorporated into diets ad libitum. Sprague-Dawley rats received experimental diets 5 groups (standard AIN 93 based control with beef tallow, extract oil 8%, and 4%, and Mortierella alpina in diet 10% and 20%) over all experiment duration (pre-mating, mating, gestation, lactation, and after weaning 4 weeks). Pups born during this period consumed same diets after wean for 4 weeks. There was no statistical significance of diet effects in reproductive performance and fertility from birth to weaning. But the groups of Mortierella alpine diet were lower of weight gain and diet intake after weaning. The serum lipids were significantly different with diet groups, higher TG in LO (oil 4%) group of dams, and higher total cholesterol in LF (M. alpina 10%) of pups, although serum albumin content was not significantly different in diet group. The spent-time and memory effect within 4 weeks of T-Morris water maze pass test in dam and 7-week- age pups did not differ in diet groups. On the count of backing error in weaning period of pups was lower in HO(extracted oil 8%) group. In the group of 10% and 20% Mortierella alpina diet, DNA content was lower in brain with lower body weight, but liver DNA relative to body weight was higher than control. Further correlation analyses would be needed DNA and arachidonic acid intakes, with Mortierella alpina diet digestion rate.

• 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.

• Tuberculosis and Respiratory Diseases
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• v.63 no.6
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• pp.497-506
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• 2007

Background: The present investigation was performed in rats and isolated human neutrophils in order to confirm the presumptive role of the positive feedback loop of cytosolic phospholipase $A_2$ ($cPLA_2$) activation by plateletactivating factor (PAF). Methods: The possible formation of the positive feedback loop of the $cPLA_2$ activation and neutrophilic respiratory burst was investigated in vivo and in vitro by measurement of the parameters denoting acute lung injury. In addition, morphological examinations and electron microscopic cytochemistry were performed for the detection of free radicals in the lung. Results: Five hours after intratracheal instillation of PAF ($5{\mu}g/rat$), the lung leak index, lung myeloperoxidase (MPO) activity, the number of neutrophils and the concentration of cytokine-induced neutrophil chemoattractant (CINC) in bronchoalveolar lavage fluid were increased by PAF as compared with those of control rats. The NBT assay and cytochrome-c reduction assay revealed an increased neutrophilic respiratory burst in isolated human neutrophils following exposure to PAF. Lung and neutrophilic $cPLA_2$ activity were increased following PAF exposure and exposure to hydrogen peroxide increased $cPLA_2$ activity in the lung. Histologically, inflammatory findings of the lung were observed after PAF treatment. Remarkably, as determined by $CeCl_3$ cytochemical electron microscopy, increased production of hydrogen peroxide was identified in the lung after PAF treatment. Conclusion: PAF mediates acute oxidative lung injury by the activation of $cPLA_2$, which may provoke the generation of free radicals in neutrophils.

• Clinical and Experimental Reproductive Medicine
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• v.27 no.1
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• pp.47-57
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• 2000

Objective: The effects of cryopreservation with or without coculture on the in vitro development of blastomere-biopsied 8-cell mouse embryos were investigated. This experimental study was originally designed for the setup of a preclinical mouse model for the preimplantation genetic diagnosis (PGD) in human. Methods: Eight-cell embryos were obtained after in vitro fertilization (IVF) from F1 hybrid mice (C57BL(표현불가)/CBA(표현불가)). Using micromanipulation, one to four blastomeres were aspirated through a hole made in the zona pellucida by zona drilling (ZD) with acid Tyrode's solution (ATS). A slow-freezing and rapid-thawing protocol with 1.5M dimethyl sulfoxide (DMSO) and 0.1M sucrose as cryoprotectant was used for the cryopreservation of blastomere- biopsied 8-cell mouse embryos. After thawing, embryos were cultured for 110 hours in Ham's F-10 supplemented with 0.4% bovine serum albumin (BSA). In the coculture group, embryos were cultured for 110 hours on the monolayer of Vero cells in the same medium. The blastocyst formation was recorded, and the embryos developed beyond blastocyst stage were stained with 10% Giemsa to count the total number of nuclei in each embryo. Results: The survival rate of embryos after cryopreservation was significantly lower in the blastomere-biopsied (7/8, 6/8, 5/8, and 4/8 embryos) groups than in the non-biopsied, zona intact (ZI) group. Without the coculture, the blastocyst formation rate of embryos after cryopreservation was not significantly different among ZI, the zona drilling only (ZD), and the balstomere-biopsied groups, but it was significantly lower than in the non-cryopreserved control group. The mean number of cells in embryos beyond blastocyst stage was significantly higher in the control group ($50.2{\pm}14.0$) than in 6/8 ($26.5{\pm}6.2$), 5/8 ($25.0{\pm}5.5$), and 4/8 ($17.8{\pm}7.8$) groups. With the coculture using Vero cells, the blastocyst formation rate of embryos after cryopreservation was significantly lower in 5/8 and 4/8 groups, compared with the control, 7/8, and 6/8 groups. The mean number of cells in embryos beyond blastocyst stage was also significantly lower in 4/8 group ($25.9{\pm}10.2$), compared with the control ($50.2{\pm}14.0$), 7/8 ($56.0{\pm}22.2$), and 6/8 ($55.3{\pm}25.5$) groups. Conclusion: After cryopreservation, blastomere-biopsied mouse embryos have a significantly impaired developmental competence in vitro, but this detrimental effect might be prevented by the coculture with Vero cells in 8-cell mouse embryos biopsied one or two blastomeres. Biopsy of mouse embryos after ZD with ATS is a safe and highly efficient preclinical model for PGD of human embryos.

• The Korean Journal of Nuclear Medicine
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• v.36 no.3
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• pp.195-202
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• 2002

Purpose: For clinical application of beta-emitter labeled antibody, high specific activity is imporiant. Carrier-free $^{188}Re$ from $^{188}W/^{188}Re$ generator is an ideal radionuclide for this purpose. However, low stability of $^{188}Re$ labeled antibody, especially in high specific activity, due to radiolytic decomposition by high energy (2.1 MeV) beta ray was problem. We studied the stability of $^{188}Re$ labeled antibody, and stabilizing effect of several stabilizers. Materials and Methods: Pre-reduced monoclonal antibody (CEA79.4) was labeled with $^{188}Re$ by incubating with generator-eluted $^{188}Re-perrhenate$ in the presence of stannous tartrate for 2 hr at room temperature. Radiochemical purity of each preparation was determined by chromatography. Human serum albumin was added to the labeled antibodies (2%). Stability of $^{188}Re-CEA79.4$ was investigated in the presence of ascorbic acid, ethanol, of Tween 80 as stabilizing agents. Results: Labeling efficiencies were $88{\pm}4%\;(n=12)$. Specific activities of $1.25{\sim}4.77MBq/{\mu}g$ were obtained. If stored after purging with $N_2$, all the preparations were stable for 10 hr. However, stability decreased in the presence of air. Perrhenate and $^{188}Re-tartrate$ was major impurity in declined preparation. colloid-formation was not a significant problem in all cases. Addition of ascorbic acid stabilized the labeled antibodies either under $N_2$ or under air by reducing the formation of perrhenate. Conclusion: High specific activity $^{188}Re$ labeled antibody is unstable, especially, in the presence of oxygen. Addition of ascorbic acid increased the stability.

• Nuclear Medicine and Molecular Imaging
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• v.41 no.4
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• pp.309-316
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• 2007

Purpose: Sentinel lymph node biopsy became the standard procedure in early breast cancer surgery. Faculty members might be exposed to a trace amount of radiation. The aim of this study is to quantify the radiation exposure and verify the safety of the procedure and the facilities, especially during pathologic process. Materials and Methods: Sentinel lymph node biopsies with Tc-99m human serum albumin were performed as routine clinical work. Exposed radiation doses were measured in pathologic technologist, nuclear medicine technologist, and nuclear medicine physician using a thermoluminescence dosimeter (TLD) during one month. We also measured the residual radioactivities or absorbed dose rates, the exposure distance and time during procedure, the radiation dose of the waste and the ambient equivalent dose of the pathology laboratory. Results: Actual exposed doses were 0.21 and 0.85 (uSv/study) for the whole body and hand of pathology technologist after 47 sentinel node pathologic preparations were performed. Whole body exposed doses of nuclear medicine physician and technologist were 0.2 and 2.3 (uSv/study). According to this data and the exposure threshold of the general population (1 mSv), at least 1100 studies were allowed in pathology technologist. The calculated exposed dose rates (${\mu}$ Sv/study) from residual radioactivities data were 2.47/ 22.4 ${\mu}$ Sv (whole body/hand) for the surgeon; 0.22/ 0 ${\mu}$ Sv for operation nurse. The ambient equivalent dose of the pathology laboratory was 0.02-0.03 mR/hr. The radiation dose of the waste was less than 100 Bq/g and nearly was not detected. Conclusion: Pathologic procedure relating sentinel lymph node biopsy using radioactive colloid is safe in terms of the radiation safety.(Nucl Med Mol Imaging 2007;41(4);309-316)

• Tuberculosis and Respiratory Diseases
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• v.39 no.6
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• pp.490-501
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• 1992

Background: Isocyanate is the most significant cause of occupational asthma in this country. The mechanism of isocyanate induced bronchoconstriction is unclear. Subjects and Method: To observe its immunologic and clinical findings, we performed methacholine bronchial challenge test (MBCT), toluene diisocyanate (TDI)-bronchoprovocation test (BPT) and RAST to TDI-, diphenylmethane diisocyanate (MDI)-, and hexamethylene diisocyanate (HDI)-human serum albumin (HSA) conjugate in 22 isocyanate-sensitive asthmatic workers. Results: BPT revealed early (11), dual (5), and late only (6) asthmatic responses. Their latent period ranged from 3 to 120 months (mean:45.9 months). Three cases (13.6%)showed a negative response on initial MBCT, but following MBeT performed 24 hours after TDI-BPT revealed the development of airway hyperresponsivenss. Twelve (54.5%) workers had increased specific IgE to TDI-HSA, seven (31.8%) had to MDI-HSA, and nine (40.9%) had to HDI-HSA conjugate. The prevalence of specific IgE was not associated with latent period, type of asthmatic responses, smoking, and atopic status. After 3 months' avoidance from workplace, airway hyperresponsiveness was improved in 10 (38.3%), among 12 followed cases. Conclusion: It is suggested that isocyanate can induce IgE-mediated bronchoconstriction in 59.1% of isocyanate-sensitive asthmatic workers. Isocyanate-induced asthma can occur even though MBeT showed a negative result, and measurement of the changes of airway hyperresponsivenss after isocyanate-BPT could be helpful to diagnose isocyanate-sensitive asthma.

• Journal of the Korean Society of Food Science and Nutrition
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• v.44 no.1
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• pp.35-43
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• 2015

In this study, hot water extract from sea buckthorn (Hippophae rhamnoides L.) leaves fermented with Hericium erinaceum mycelium (SBT-HE) was assessed for protection against oxidative modification of biological macromolecules and cell death. Antioxidant activity of SBT-HE was evaluated based on 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical, 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical, and peroxyl radical scavenging assays. SBT-HE showed 65.06% DPPH radical scavenging activity at $500{\mu}g/mL$, 98.83% ABTS radical scavenging activity at $50{\mu}g/mL$, and 44.03% peroxyl radical scavenging activity at $100{\mu}g/mL$. SBT-HE significantly inhibited DNA strand breakage induced by peroxyl radical. SBT-HE also prevented peroxyl radical-mediated human serum albumin modification. SBT-HE effectively inhibited $H_2O_2$-induced cell death and significantly increased cell survival by 21.59% at $100{\mu}g/mL$. SBT-HE also reduced intracellular reactive oxygen species levels in $H_2O_2$-treated cells. The results suggest that SBT-HE can contribute to antioxidant activity and protect cells from oxidative stress-induced cell injury.